1 00:00:07,040 --> 00:00:12,760 I WANT TO INTRODUCE MY 2 00:00:12,760 --> 00:00:14,400 COLLEAGUE, WHO HELPED ME WITH A 3 00:00:14,400 --> 00:00:16,040 LOT OF THIS, SHE'S GOING TO 4 00:00:16,040 --> 00:00:18,600 INTRODUCE ALL THE SPEAKERS AS 5 00:00:18,600 --> 00:00:21,880 THEY COME UP. 6 00:00:21,880 --> 00:00:23,280 7 00:00:23,280 --> 00:00:25,080 >> THANK YOU FOR COMING. 8 00:00:25,080 --> 00:00:26,360 BEFORE WE START, I WANT TO 9 00:00:26,360 --> 00:00:28,320 MENTION TOMORROW MORNING WE HAVE 10 00:00:28,320 --> 00:00:35,120 A POSTER SESSION AT THE FAES 11 00:00:35,120 --> 00:00:39,960 TERRACE WITH REFRESHMENTS 12 00:00:39,960 --> 00:00:47,040 FOLLOWED BY WALS LECTURE WITH 13 00:00:47,040 --> 00:00:49,560 DR. IRENE GEORGAKODI. 14 00:00:49,560 --> 00:00:52,400 WE'RE EXCITED TO INTRODUCE AN 15 00:00:52,400 --> 00:00:57,640 AWESOME SPEAKER, DR. KAITLIN 16 00:00:57,640 --> 00:01:00,520 SADTLER, CHIEF OF SECTION 4 AT 17 00:01:00,520 --> 00:01:02,560 NIBIB, HER LAB FOCUSES ON 18 00:01:02,560 --> 00:01:06,320 DEVELOPMENT OF IMMUNE ACTIVE 19 00:01:06,320 --> 00:01:09,240 BIOMATERIALS FOR REGENERATIVE 20 00:01:09,240 --> 00:01:09,880 MEDICINE USING MECHANISM-BASED 21 00:01:09,880 --> 00:01:15,160 NODOLOGY AND AND -- METHODOLOGR 22 00:01:15,160 --> 00:01:19,840 OUR OPENING TALK. 23 00:01:19,840 --> 00:01:20,120 [APPLAUSE] 24 00:01:20,120 --> 00:01:22,600 >>THANK YOU. 25 00:01:22,600 --> 00:01:24,040 THANKS FOR THE NICE 26 00:01:24,040 --> 00:01:26,640 INTRODUCTION, ALSO, STEVE, FOR 27 00:01:26,640 --> 00:01:28,080 THE EFFORT OF ORGANIZING THIS 28 00:01:28,080 --> 00:01:28,400 TOGETHER. 29 00:01:28,400 --> 00:01:31,920 I KNOW IT'S NO SMALL FEAT. 30 00:01:31,920 --> 00:01:37,920 HUGE THANK YOU FOR THE 31 00:01:37,920 --> 00:01:39,360 INVITATION, A I'M A TENURE-TRACK 32 00:01:39,360 --> 00:01:40,440 INVESTIGATOR AT NIBIB PRESENTING 33 00:01:40,440 --> 00:01:42,720 ON WORK OUR LAB HAS BEEN DOING 34 00:01:42,720 --> 00:01:45,880 IN THE PAST FEW YEARS ON MUSCLE 35 00:01:45,880 --> 00:01:48,400 TRAUMA AND THE WAY IMMUNE CELLS 36 00:01:48,400 --> 00:01:50,240 ARE IMPORTANT AND REACTING TO 37 00:01:50,240 --> 00:01:51,920 MATERIALS WE PUT IN AND 38 00:01:51,920 --> 00:01:58,280 SUBSEQUENT EFFECT ON TISSUE 39 00:01:58,280 --> 00:01:59,120 GROWTH. 40 00:01:59,120 --> 00:02:00,800 AS A BIT OF A BACKGROUND AS TO 41 00:02:00,800 --> 00:02:02,520 WHERE I CAME FROM AND HOW I 42 00:02:02,520 --> 00:02:04,400 ENDED UP BACK HERE AT THE NIH AS 43 00:02:04,400 --> 00:02:06,520 A P.I., I'M A LOCAL. 44 00:02:06,520 --> 00:02:10,040 I WENT THROUGH THE FREDERICK 45 00:02:10,040 --> 00:02:11,760 COUNTY PUBLIC SCHOOL SYSTEM 46 00:02:11,760 --> 00:02:18,440 PRIOR TO DEGREE AT UMBC, A YEAR 47 00:02:18,440 --> 00:02:20,400 AT POSTBAC BEFORE MY Ph.D. AT 48 00:02:20,400 --> 00:02:24,560 JOHNS HOPKINS TELLING MYSELF I 49 00:02:24,560 --> 00:02:26,640 COULD NOT LIVE IN MARYLAND AND 50 00:02:26,640 --> 00:02:28,960 ONLY MARYLAND, POPPED TO M.I.T., 51 00:02:28,960 --> 00:02:31,680 POSTDOC, FOR RETURNING AS A 52 00:02:31,680 --> 00:02:32,480 STADTMAN INVESTIGATOR AT NIBIB, 53 00:02:32,480 --> 00:02:39,440 FALL OF 2019 GETTING MY LAB 54 00:02:39,440 --> 00:02:41,400 STARTED, EARLY 2020, A BRILLIANT 55 00:02:41,400 --> 00:02:43,520 TIME TO OPEN THE LAB. 56 00:02:43,520 --> 00:02:48,480 THE MICS ALSO DON'T LIKE COVID. 57 00:02:48,480 --> 00:02:52,240 UP FRONT, QUICK ACKNOWLEDGMENTS. 58 00:02:52,240 --> 00:02:53,880 WE'RE GOOD? 59 00:02:53,880 --> 00:02:55,040 QUICK ACKNOWLEDGMENTS. 60 00:02:55,040 --> 00:02:58,600 I'M A NEW P.I., AS A NEW P.I. 61 00:02:58,600 --> 00:03:00,240 I'M BUILDING THE LAB, AS ALWAYS, 62 00:03:00,240 --> 00:03:01,880 A HUGE TEAM EFFORT, SO ANYTHING 63 00:03:01,880 --> 00:03:04,640 DONE HERE I DID MOST OF THE 64 00:03:04,640 --> 00:03:05,400 PAPERWORK ON, THE FOLKS IN THE 65 00:03:05,400 --> 00:03:07,920 PHOTO ARE THE ONES THAT DID MOST 66 00:03:07,920 --> 00:03:11,200 OF THE PIPETTING. 67 00:03:11,200 --> 00:03:15,120 HUGE THANKS TO EVERYBODY 68 00:03:15,120 --> 00:03:21,280 INVOLVED, ESPECIALLY DR. 69 00:03:21,280 --> 00:03:26,080 LACKWANI, AND ANOTHER 70 00:03:26,080 --> 00:03:27,600 POSTDOCTORAL FELLOW AND ANOTHER 71 00:03:27,600 --> 00:03:38,160 MD/PHD STUDENT, AND WE ALSO HAVE 72 00:03:39,840 --> 00:03:42,080 SABRINA, A TECHNICIAN, AN 73 00:03:42,080 --> 00:03:45,440 INDEPENDENT RESEARCH SCHOLAR, 74 00:03:45,440 --> 00:03:47,960 AND A POSTBAC NOW AN MD/PHD 75 00:03:47,960 --> 00:03:49,040 STUDENT AS WELL. 76 00:03:49,040 --> 00:03:50,240 THESE FOLKS AND FOLKS THAT 77 00:03:50,240 --> 00:03:52,200 AREN'T IN THIS PICTURE THAT I'LL 78 00:03:52,200 --> 00:03:54,080 MENTION LATER HELPED BUILD THE 79 00:03:54,080 --> 00:03:56,360 LAB, SO MANY THANKS TO THEM. 80 00:03:56,360 --> 00:03:59,200 SO, JUST AS A QUICK OVERVIEW IN 81 00:03:59,200 --> 00:04:01,280 TERMS OF WHAT WE THINK OF WHEN 82 00:04:01,280 --> 00:04:03,240 WE'RE TALKING ABOUT IMMUNE 83 00:04:03,240 --> 00:04:05,560 RESPONSES TO TRAUMATIC INJURY 84 00:04:05,560 --> 00:04:06,880 AND BIOMATERIALS, PREVIOUSLY THE 85 00:04:06,880 --> 00:04:08,080 IMMUNE SYSTEM WAS VIEWED AS A 86 00:04:08,080 --> 00:04:12,480 BIT OF A BADDIE WHEN IT CAME TO 87 00:04:12,480 --> 00:04:13,040 IMPLANTED MATERIALS, BUT 88 00:04:13,040 --> 00:04:14,800 ULTIMATELY THERE'S GOOD AND BAD 89 00:04:14,800 --> 00:04:17,520 THINGS THAT HAPPEN. 90 00:04:17,520 --> 00:04:19,480 IF YOU THINK OF BIOMATERIAL 91 00:04:19,480 --> 00:04:20,800 SCAFFOLD, WHERE WE WANT THEM TO 92 00:04:20,800 --> 00:04:22,200 REMODEL AND INCORPORATE INTO THE 93 00:04:22,200 --> 00:04:23,520 TISSUE, THE IMMUNE SYSTEM PLAYS 94 00:04:23,520 --> 00:04:25,600 AN IMPORTANT ROLE THERE, THE 95 00:04:25,600 --> 00:04:26,840 LOCATION OF THAT IMPLANTATION, 96 00:04:26,840 --> 00:04:28,800 PRESENCE OF INJURY, THE TYPE OF 97 00:04:28,800 --> 00:04:29,880 MATERIAL WE IMPLANT CHANGES THE 98 00:04:29,880 --> 00:04:31,840 TYPES OF IMMUNE CELLS THAT COME 99 00:04:31,840 --> 00:04:32,400 IN. 100 00:04:32,400 --> 00:04:38,240 THAT CAN ALSO THEN AFFECT THEIR 101 00:04:38,240 --> 00:04:39,440 ACTIVATION AND POLARIZATION, 102 00:04:39,440 --> 00:04:40,280 CREATING SCAFFOLD IMMUNE 103 00:04:40,280 --> 00:04:44,680 ENVIRONMENT AND ALTER STEM CELL 104 00:04:44,680 --> 00:04:52,320 DIVERSITY, 105 00:04:52,320 --> 00:04:53,520 -- DIFFERENTIATION WITH 106 00:04:53,520 --> 00:04:54,080 DIFFERENT OUTCOMES. 107 00:04:54,080 --> 00:04:57,120 IF WE THINK OF MEDICAL DEVICES, 108 00:04:57,120 --> 00:04:57,960 COSMETIC IMPLANT NOT NECESSARILY 109 00:04:57,960 --> 00:04:59,560 MEANT TO INTEGRATE WITH 110 00:04:59,560 --> 00:05:02,360 SURROUNDING TISSUE BUT WORK MORE 111 00:05:02,360 --> 00:05:05,680 IN THE STANDALONE WAY, WE HAVE 112 00:05:05,680 --> 00:05:07,000 THE CANONICAL FOREIGN BODY 113 00:05:07,000 --> 00:05:09,080 RESPONSE, IN THIS BOTTOM RIGHT 114 00:05:09,080 --> 00:05:09,480 HERE. 115 00:05:09,480 --> 00:05:11,840 WITH THAT, WHAT WE SEE IS IF YOU 116 00:05:11,840 --> 00:05:14,960 THINK OF A BIG PLASTIC DEVICE, 117 00:05:14,960 --> 00:05:17,040 WE HAVE PROTEIN ABSORPTION, 118 00:05:17,040 --> 00:05:18,600 IMMUNE CELLS TRYING TO 119 00:05:18,600 --> 00:05:20,880 INTEGRATE, THEY CAN'T, SO WHEN 120 00:05:20,880 --> 00:05:30,200 THEY CAN'T DEGRADE IT WE GET 121 00:05:30,200 --> 00:05:31,200 FIBROTIC CAPSULATION. 122 00:05:31,200 --> 00:05:33,760 I WAS ABLE TO COLLABORATE WITH 123 00:05:33,760 --> 00:05:39,600 AN ARTIST WHO PUT TOGETHER A 124 00:05:39,600 --> 00:05:43,000 VIDEO, THE ARTIST IS MORE 125 00:05:43,000 --> 00:05:45,400 TALENTED THAN I, HILARIOUS 126 00:05:45,400 --> 00:05:45,600 VIDEO. 127 00:05:45,600 --> 00:05:48,840 THERE IS THIS LARGE DISTURBANCE 128 00:05:48,840 --> 00:05:50,160 IN HOMEOSTASIS, AND THIS 129 00:05:50,160 --> 00:05:51,680 PERSISTENT INFLAMMATION, SO HOW 130 00:05:51,680 --> 00:05:54,520 DO OUR CELLS NOT START TO ATTACK 131 00:05:54,520 --> 00:05:57,280 ITSELF, SO IF WE HAVE CONSTANT 132 00:05:57,280 --> 00:05:57,880 INFLAMMATION AROUND DAMAGED 133 00:05:57,880 --> 00:05:59,320 TISSUE HOW COME THAT SWITCH 134 00:05:59,320 --> 00:06:00,880 DOESN'T HAPPEN AND WE GET A 135 00:06:00,880 --> 00:06:03,520 NEGATIVE RESPONSE? 136 00:06:03,520 --> 00:06:06,760 SO OUR GOAL TO TO VIEW 137 00:06:06,760 --> 00:06:07,760 ENGINEERING AS AN IMMUNOLOGIST, 138 00:06:07,760 --> 00:06:12,720 AS MENTIONED, SO WE'RE USING 139 00:06:12,720 --> 00:06:13,480 THIS MECHANISTIC BIOLOGY TO 140 00:06:13,480 --> 00:06:14,880 OUNCE MECHANISMS THAT OF GO EARN 141 00:06:14,880 --> 00:06:16,080 IMMUNE RESPONSES TO MATERIALS 142 00:06:16,080 --> 00:06:17,520 AND SUBSEQUENT EFFECT ON TISSUE 143 00:06:17,520 --> 00:06:18,520 GROWTH AND DEVELOPMENT. 144 00:06:18,520 --> 00:06:22,040 THAT WAY WE CAN IDENTIFY TARGETS 145 00:06:22,040 --> 00:06:22,600 FOR RATIONALLY DESIGNED 146 00:06:22,600 --> 00:06:23,920 THERAPEUTICS, BUILT FROM THE 147 00:06:23,920 --> 00:06:24,480 BOTTOM UP. 148 00:06:24,480 --> 00:06:32,080 SO, IN TERMS OF MODELS, OUR 149 00:06:32,080 --> 00:06:37,720 LABORATORY USES VML MOD MODEL A 150 00:06:37,720 --> 00:06:40,960 MOUSE, 30 MILLIGRAMS OF TISSUE 151 00:06:40,960 --> 00:06:41,640 REMOVED BILATERALLY FROM 152 00:06:41,640 --> 00:06:43,280 QUADRICEPS, RESULTS IN 30% 153 00:06:43,280 --> 00:06:44,400 PERMANENT DEFECT IN MUSCLE 154 00:06:44,400 --> 00:06:45,600 FUNCTION. 155 00:06:45,600 --> 00:06:47,000 IN TERMS OF THE DIFFERENT 156 00:06:47,000 --> 00:06:48,200 MATERIALS THAT WE IMPLANT WE 157 00:06:48,200 --> 00:06:50,520 LEAVE THE INJURY ALONE FOR 158 00:06:50,520 --> 00:06:53,160 CONTROL BUT ALSO HAVE CHOSEN TWO 159 00:06:53,160 --> 00:06:56,640 POLES OF IMMUNE RESPONSE TO 160 00:06:56,640 --> 00:06:58,680 MATERIALS, A POWDERED 161 00:06:58,680 --> 00:06:59,840 DECELLULARRIZED EXTRACELLULAR 162 00:06:59,840 --> 00:07:02,440 MATRIX, AND A POWDERED 163 00:07:02,440 --> 00:07:05,920 POLYETHYLENE, OUR PRO FIBROTIC 164 00:07:05,920 --> 00:07:06,200 MATERIAL. 165 00:07:06,200 --> 00:07:10,200 ECM IS USED FOR TREATMENT OF 166 00:07:10,200 --> 00:07:12,080 SKIN WOUNDS, HERNIAS, ALSO HAS 167 00:07:12,080 --> 00:07:16,680 GONE UNDERCLINICAL TRIALS FOR 168 00:07:16,680 --> 00:07:18,200 VOLUMETRIC INJURIES, IT'S USED 169 00:07:18,200 --> 00:07:19,600 IN ORTHOPEDIC IMPLANTS BUT WHEN 170 00:07:19,600 --> 00:07:20,720 YOU GET THOSE SMALL WEAR 171 00:07:20,720 --> 00:07:22,480 PARTICLES AND THOSE WIND UP 172 00:07:22,480 --> 00:07:24,120 EMBEDDING IN LOCAL TISSUE YOU 173 00:07:24,120 --> 00:07:25,840 GET STRONG INFLAMMATION AND 174 00:07:25,840 --> 00:07:28,840 FIBROSIS, AND IT'S REALLY A 175 00:07:28,840 --> 00:07:29,160 NASTY EFFECT. 176 00:07:29,160 --> 00:07:32,000 SO, WHEN WE GOT KICK STARTED, WE 177 00:07:32,000 --> 00:07:34,280 WOUND UP DEVELOPING A FEW FLOW 178 00:07:34,280 --> 00:07:36,280 CYTOMETRY PANELS TO EVALUATE THE 179 00:07:36,280 --> 00:07:37,480 DIFFERENT IMMUNE CELLS COMING IN 180 00:07:37,480 --> 00:07:38,800 AND RESPONDING TO THE INJURY AND 181 00:07:38,800 --> 00:07:40,080 TO THE MATERIAL. 182 00:07:40,080 --> 00:07:41,720 I WON'T GET INTO ALL OF THE 183 00:07:41,720 --> 00:07:44,960 CELLS THAT WE LOOKED AT, BUT WE 184 00:07:44,960 --> 00:07:46,880 WERE ABLE TO ANALYZE THESE I 185 00:07:46,880 --> 00:07:49,160 HAVE A STANDARD MANUAL GAIN AND 186 00:07:49,160 --> 00:07:50,160 APPLYING COMPUTATIONAL METHODS 187 00:07:50,160 --> 00:07:55,200 AS WELL, AND IF YOU WANT TO 188 00:07:55,200 --> 00:07:56,400 AVOID THE TROUBLESHOOTING WE 189 00:07:56,400 --> 00:07:58,640 HAVE A COMMON ARTICLE IN NATURE 190 00:07:58,640 --> 00:07:59,720 REVIEWS METHODS IN COLLABORATION 191 00:07:59,720 --> 00:08:03,440 WITH THE WOLFE LAB FROM NCI ON 192 00:08:03,440 --> 00:08:06,600 SOME TIPS TO LOOK OUT FOR WHILE 193 00:08:06,600 --> 00:08:07,600 DESIGNING THESE PANELS BECAUSE 194 00:08:07,600 --> 00:08:11,160 THEY WERE PAINFUL WHEN YOU GET 195 00:08:11,160 --> 00:08:11,840 HIGHLY AUTOFLUORESCENT SAMPLES. 196 00:08:11,840 --> 00:08:14,280 SO, SINCE WE WERE INTERESTED IN 197 00:08:14,280 --> 00:08:16,120 THIS ASPECT OF AUTOIMMUNITY WHAT 198 00:08:16,120 --> 00:08:18,400 WE WOUND UP DIGGING INTO HERE IN 199 00:08:18,400 --> 00:08:21,560 TERMS OF THE MYELOID IMMUNE 200 00:08:21,560 --> 00:08:22,680 CELLS WERE THE 201 00:08:22,680 --> 00:08:23,760 ANTIGEN-PRESENTING CELLS, 202 00:08:23,760 --> 00:08:23,960 APCs. 203 00:08:23,960 --> 00:08:26,200 DENDRITIC CELLS IN TERMS OF WHAT 204 00:08:26,200 --> 00:08:29,240 COMES INTO THIS INJURY SPACE, 205 00:08:29,240 --> 00:08:32,440 THEY PEAK AND COUNT BY ABOUT 206 00:08:32,440 --> 00:08:34,600 SEVEN DAYS POST INJURY AND 207 00:08:34,600 --> 00:08:36,720 QUICKLY CLEAR, PERSIST IN LOW 208 00:08:36,720 --> 00:08:39,640 NUMBERS THROUGH 42 DAYS 209 00:08:39,640 --> 00:08:40,080 POST-INJURY. 210 00:08:40,080 --> 00:08:42,800 SO, WHAT WE WIND UP SEEING IS 211 00:08:42,800 --> 00:08:45,000 THAT ECM TREATMENTS, SO 212 00:08:45,000 --> 00:08:45,680 TREATMENT WITH THIS 213 00:08:45,680 --> 00:08:46,960 PRO-REGENERATIVE MATERIAL WHICH 214 00:08:46,960 --> 00:08:55,880 YOU CAN SEE IN THE TEAL COLOR, 215 00:08:55,880 --> 00:08:57,200 POPULATION OF CONVENTIONAL 216 00:08:57,200 --> 00:08:58,200 DENDRITIC CELL TYPE ONES. 217 00:08:58,200 --> 00:09:01,560 SORRY ABOUT THE NAMES, IT'S HOW 218 00:09:01,560 --> 00:09:01,920 IMMUNOLOGY GOES. 219 00:09:01,920 --> 00:09:04,200 I LEARN THEM ON A DAILY BASIS. 220 00:09:04,200 --> 00:09:11,080 WE ENRICHED THIS POPULATION THAT 221 00:09:11,080 --> 00:09:15,040 EXPRESS HIGH LEVELS, LACKED 222 00:09:15,040 --> 00:09:17,320 CANONICAL RECEPTOR. 223 00:09:17,320 --> 00:09:19,160 THESE ARE ASSOCIATED WITH 224 00:09:19,160 --> 00:09:20,840 ANTIGEN CROSS-PRESENTATION 225 00:09:20,840 --> 00:09:24,760 CHARACTERIZED BY TWO MARKERS, 226 00:09:24,760 --> 00:09:26,640 EXTERIOR ONE AND CD103, NOT SO 227 00:09:26,640 --> 00:09:28,680 MUCH WITH POLYETHYLENE 228 00:09:28,680 --> 00:09:28,960 TREATMENT. 229 00:09:28,960 --> 00:09:30,360 IF WE LOOK IN THE LITERATURE 230 00:09:30,360 --> 00:09:37,160 ABOUT OTHER MARKERS EXPRESSED ON 231 00:09:37,160 --> 00:09:41,920 THEM DENDRITIC CELLS INTEREST 232 00:09:41,920 --> 00:09:44,040 BEEN DESCRIBED, Th2 IMMUNITY 233 00:09:44,040 --> 00:09:47,320 IS IMPORTANT, AND ANTIGEN 234 00:09:47,320 --> 00:09:49,600 CROSS-PRESENTATION AND CD8 AND 235 00:09:49,600 --> 00:09:54,040 INDUCE THE TREGS BEEN IMPLICATED 236 00:09:54,040 --> 00:09:56,640 IN TOLERANCE IN A MOUSE MODEL BY 237 00:09:56,640 --> 00:09:58,760 MARK DAVIS' GROUP, WE'RE GETTING 238 00:09:58,760 --> 00:10:03,680 CURIOUS WHETHER THEY PLAY AN 239 00:10:03,680 --> 00:10:04,560 IMMUNOREGULATORY ROLE. 240 00:10:04,560 --> 00:10:08,160 WE'RE GOING TO FOCUS MOSTLY ON 241 00:10:08,160 --> 00:10:09,440 THIS ECM TREATMENT, 242 00:10:09,440 --> 00:10:10,240 PRO-REGENERATIVE MATERIAL. 243 00:10:10,240 --> 00:10:12,080 FURTHERMORE WHEN WE LOOK AT 244 00:10:12,080 --> 00:10:15,000 MATERIAL TYPE OR THE DIFFERENT 245 00:10:15,000 --> 00:10:16,920 FORMS OF MATERIAL, IN ADDITION 246 00:10:16,920 --> 00:10:20,480 TO POWDERED MATERIAL IF WE PLACE 247 00:10:20,480 --> 00:10:23,920 ECM SHEETS, USED IN THE CLINIC, 248 00:10:23,920 --> 00:10:26,240 AS WELL AS LARGER POLYETHYLENE 249 00:10:26,240 --> 00:10:28,400 PARTICLES WE SEE THE SAME 250 00:10:28,400 --> 00:10:28,640 PATTERN. 251 00:10:28,640 --> 00:10:31,400 SO IT SEEMS TO BE AGNOSTIC TO 252 00:10:31,400 --> 00:10:32,920 THAT FORMULATION OF THE 253 00:10:32,920 --> 00:10:34,840 MATERIAL. 254 00:10:34,840 --> 00:10:35,840 SO IT STILL MAINTAINS THE SAME 255 00:10:35,840 --> 00:10:45,600 PATTERN WHERE THE SHEETZ ARE 256 00:10:45,600 --> 00:10:46,560 RECRUITING DENDRITIC CELLS, PE 257 00:10:46,560 --> 00:10:47,640 PARTICLES ARE NOT. 258 00:10:47,640 --> 00:10:49,520 WE WANTED TO KNOW WHETHER OR NOT 259 00:10:49,520 --> 00:10:50,920 THEY WERE THESE CROSS-PRESENTING 260 00:10:50,920 --> 00:10:52,360 DENDRITIC CELLS IN THE 261 00:10:52,360 --> 00:10:57,840 LITERATURE, SO WHAT WE LOOKED AT 262 00:10:57,840 --> 00:11:00,240 WAS A TRANSCRIPTION FACTOR 263 00:11:00,240 --> 00:11:03,520 REQUIRED FOR MATURATION OF CDC 264 00:11:03,520 --> 00:11:03,680 1s. 265 00:11:03,680 --> 00:11:07,240 GOOD DATA DAY, WHEN WE LOOK IN A 266 00:11:07,240 --> 00:11:08,200 THREE KNOCKOUT MICE IN RED, WE 267 00:11:08,200 --> 00:11:09,160 LOSE THEM. 268 00:11:09,160 --> 00:11:13,800 THESE DO APPEAR TO BE SAME 269 00:11:13,800 --> 00:11:15,120 CROSS-PRESENTING DEPENDENT CD1S 270 00:11:15,120 --> 00:11:16,840 IN THE LITERATURE, JUST A LITTLE 271 00:11:16,840 --> 00:11:19,400 WEIRD, WHICH IS EVERY IMMUNE 272 00:11:19,400 --> 00:11:20,000 CELL. 273 00:11:20,000 --> 00:11:27,480 NEXT QUESTION WAS ABOUT ADAPTIVE 274 00:11:27,480 --> 00:11:29,880 IMMUNITY, SO WE CHARACTERIZED 275 00:11:29,880 --> 00:11:31,440 THE IMMUNE INFILTRATE FOCUSING 276 00:11:31,440 --> 00:11:32,400 ADAPTIVE IMMUNE CELLS WITH 277 00:11:32,400 --> 00:11:33,720 ANOTHER FLOW PANEL. 278 00:11:33,720 --> 00:11:38,200 WE'VE KNOWN IN THE PAST, MY 279 00:11:38,200 --> 00:11:40,720 THESIS WORK WAS FOCUSED ON 280 00:11:40,720 --> 00:11:42,680 HELPER T CELLS AND WANTED TO 281 00:11:42,680 --> 00:11:45,680 KNOW THE LYMPHOID PROFILE AND 282 00:11:45,680 --> 00:11:46,840 POTENTIAL OF REGULATORY IMMUNE 283 00:11:46,840 --> 00:11:48,200 CELLS TO CALM THE IMMUNE 284 00:11:48,200 --> 00:11:48,560 RESPONSE. 285 00:11:48,560 --> 00:11:51,160 LOOKING AT THE DIFFERENT TYPES 286 00:11:51,160 --> 00:11:52,360 OF ADAPTIVE IMMUNE CELLS PRESENT 287 00:11:52,360 --> 00:11:54,640 YOU SEE HIGH LEVELS OF BOTH 288 00:11:54,640 --> 00:11:57,240 ALPHA BETA T CELLS AS REPORTED 289 00:11:57,240 --> 00:11:58,920 PREVIOUSLY BY OUR LAB AND 290 00:11:58,920 --> 00:12:01,440 OTHERS, AS WELL AS HIGH LEVELS 291 00:12:01,440 --> 00:12:04,920 OF NATURAL KILLER CELLS IN ECM 292 00:12:04,920 --> 00:12:05,320 TREATMENT. 293 00:12:05,320 --> 00:12:07,760 THIS IS INTERESTING BECAUSE 294 00:12:07,760 --> 00:12:10,320 NATURAL KILLER CELLS SECRETE 295 00:12:10,320 --> 00:12:20,800 CHEMOKINE THAT RECRUITS CDC 296 00:12:22,400 --> 00:12:22,840 1s. 297 00:12:22,840 --> 00:12:28,360 WE ALSO SEE THAT THESE -- THERE 298 00:12:28,360 --> 00:12:34,360 ARE CD8s EXPRESSING HELIOS, 299 00:12:34,360 --> 00:12:35,480 WE'RE PURSUING THAT FURTHER. 300 00:12:35,480 --> 00:12:37,000 WHEN WE LOOK AT GENE EXPRESSION 301 00:12:37,000 --> 00:12:39,640 OF THE T CELLS IN THE WOUND 302 00:12:39,640 --> 00:12:42,040 VERSUS DRAINING LYMPH NODE OF 303 00:12:42,040 --> 00:12:44,760 THE SAME ANIMAL WE SEE THAT 304 00:12:44,760 --> 00:12:55,320 THESE T CELLS IN LOCAL AREA HAVE 305 00:12:56,400 --> 00:12:58,000 HIGHER LEVELS OF IL, NOTHING 306 00:12:58,000 --> 00:12:59,560 EVER FITS ON A BEAUTIFUL BINARY 307 00:12:59,560 --> 00:13:02,400 SCALE BUT WE DO SEE THAT 308 00:13:02,400 --> 00:13:03,880 ENRICHMENT OF REGULATORY IL-10 309 00:13:03,880 --> 00:13:06,200 MARKER IN THE LOCAL TISSUE. 310 00:13:06,200 --> 00:13:07,960 NOW, WHEN WE'RE TALKING ABOUT 311 00:13:07,960 --> 00:13:09,800 TREATMENT SPECIFICALLY AND LOOK 312 00:13:09,800 --> 00:13:15,080 AT THE LYMPH NODE WE CAN SEE 313 00:13:15,080 --> 00:13:17,040 HIGH UPREGULATION OF -- THIS IS 314 00:13:17,040 --> 00:13:18,480 THE MUSCLE TISSUE IN COMPARISON 315 00:13:18,480 --> 00:13:20,960 TO OTHER TREATMENT GROUPS. 316 00:13:20,960 --> 00:13:23,800 WE CAN SEE ETM TREATMENT DOES 317 00:13:23,800 --> 00:13:27,400 UPREGULATE IL-HAD AND IL-13, 318 00:13:27,400 --> 00:13:30,400 AGAIN CANONICAL Th2 CYTOKINES, 319 00:13:30,400 --> 00:13:33,240 IL-10 AND IL-2RA FOR T-CELL 320 00:13:33,240 --> 00:13:35,200 ACTIVATION AND ODDLY AIR, TOSSED 321 00:13:35,200 --> 00:13:41,520 IN FOR FINE LIKE ANYONE DOES, 322 00:13:41,520 --> 00:13:42,120 AUTOIMMUNE REGULATORY ELEMENT, 323 00:13:42,120 --> 00:13:46,240 FOR T CELL DEVELOPMENT AND 324 00:13:46,240 --> 00:13:47,040 SELF-TOLERANCE, GOES UP 325 00:13:47,040 --> 00:13:48,120 PERIPHERALLY WHICH IS WEIRD. 326 00:13:48,120 --> 00:13:49,640 WE DON'T KNOW WHAT WE'RE DOING 327 00:13:49,640 --> 00:13:51,800 BUT IT HAPPENS. 328 00:13:51,800 --> 00:13:53,760 SO, INTERESTING ASSOCIATION WITH 329 00:13:53,760 --> 00:13:55,400 KIND OF SOME TOLERANCE. 330 00:13:55,400 --> 00:13:58,800 SO, NOW WE'VE TALKED ABOUT THE 331 00:13:58,800 --> 00:13:59,920 APCs, ADAPTIVE IMMUNE SYSTEM, 332 00:13:59,920 --> 00:14:01,640 WHAT ABOUT CONNECTION BETWEEN 333 00:14:01,640 --> 00:14:03,960 THE TWO? 334 00:14:03,960 --> 00:14:07,360 WHAT HAPPENS WITH IN TERMS OF 335 00:14:07,360 --> 00:14:08,560 REGULATING INACTIVATION AND 336 00:14:08,560 --> 00:14:10,520 POLARIZATION OF IMMUNE CELLS 337 00:14:10,520 --> 00:14:11,160 AFTER INJURY? 338 00:14:11,160 --> 00:14:15,960 SO WE TOOK A LOOK BACK AT THE 339 00:14:15,960 --> 00:14:17,440 BATTA 3 KNOCKOUT MODELS, T CELLS 340 00:14:17,440 --> 00:14:19,800 IN TRAINING LYMPH NODE SEVEN 341 00:14:19,800 --> 00:14:23,320 DAYS AFTER INJURY, RED BARS ARE 342 00:14:23,320 --> 00:14:26,160 THE BATA 3 KNOCKOUT MOUSE, 343 00:14:26,160 --> 00:14:27,120 INCREASE IN T-CELL ACTIVATION 344 00:14:27,120 --> 00:14:29,320 AND SEE THAT IN THE BLOOD. 345 00:14:29,320 --> 00:14:31,320 THIS IS BASED OFF THE EXPRESSION 346 00:14:31,320 --> 00:14:34,920 OF CD44 AND LACK OF CD62 LIGAND. 347 00:14:34,920 --> 00:14:38,520 IF WE LOOK AT MACROPHAGES, 348 00:14:38,520 --> 00:14:39,840 PREVIOUSLY IMPLICATED AS AN 349 00:14:39,840 --> 00:14:41,720 IMPORTANT PLAYER IN SCAFFOLD WE 350 00:14:41,720 --> 00:14:42,920 MODELING AND WOUND HEALING WE 351 00:14:42,920 --> 00:14:46,520 CAN SEE THEY ARE ALSO DISRUPTED 352 00:14:46,520 --> 00:14:49,160 IN THE BATF3 KNOCKOUT MODEL, 353 00:14:49,160 --> 00:14:51,240 WEIRD THINGS HAPPENING WITH 354 00:14:51,240 --> 00:14:51,520 MACROPHAGES. 355 00:14:51,520 --> 00:14:55,400 WHAT WE GET IS WE HAVE AN 356 00:14:55,400 --> 00:14:58,680 INITIAL LAG IN MACROPHAGE 357 00:14:58,680 --> 00:15:00,120 RECRUITMENT THAT PICKS UP, AND 358 00:15:00,120 --> 00:15:01,600 MORE RECRUITED TO MUSCLE 359 00:15:01,600 --> 00:15:07,320 INJURIES AT 21 DAYS IN THE BATF3 360 00:15:07,320 --> 00:15:13,240 KNOCKOUT MODEL, DIFFERENT 361 00:15:13,240 --> 00:15:15,200 SURFACE MODELINGS AND LOSE A 362 00:15:15,200 --> 00:15:17,480 MACROPHAGE MARKER IN MICE, I HAD 363 00:15:17,480 --> 00:15:20,000 TO GOOGLE WHAT IT DID, FIRST 364 00:15:20,000 --> 00:15:21,520 PAPER WAS ASSOCIATED WITH 365 00:15:21,520 --> 00:15:23,840 TOLERANCE IN MICE, ALSO GOOD 366 00:15:23,840 --> 00:15:28,680 DATA TO HAVE. 367 00:15:28,680 --> 00:15:30,720 CD301B MINUTEALLY CHANGED. 368 00:15:30,720 --> 00:15:33,360 CD206 HAS BEEN UTILIZED AS A 369 00:15:33,360 --> 00:15:35,200 MARKER OF KIND OF, YOU KNOW, 370 00:15:35,200 --> 00:15:39,280 MORE HELPFUL MACROPHAGES IN THE 371 00:15:39,280 --> 00:15:40,960 SCAFFOLD REMODELING PROCESS, 372 00:15:40,960 --> 00:15:50,040 CANCER LITERATURE ASSOCIATED 373 00:15:50,040 --> 00:15:50,800 WITH PHAGOCYTOSIS. 374 00:15:50,800 --> 00:15:52,640 OUR QUESTION IS WHAT'S BRINGING 375 00:15:52,640 --> 00:15:55,280 THEM THERE? SO WE START TO LOOK 376 00:15:55,280 --> 00:15:59,640 AT THE TWO RECEPTORS THAT 377 00:15:59,640 --> 00:16:05,960 CHARACTERIZE THESE CELLS, CD103 378 00:16:05,960 --> 00:16:06,440 103 379 00:16:06,440 --> 00:16:08,040 BINDS E-CADHERIN. 380 00:16:08,040 --> 00:16:11,240 WE LOOKED AT EXPRESSION AFTER 381 00:16:11,240 --> 00:16:12,440 INJURY, PCR AND 382 00:16:12,440 --> 00:16:14,280 IMMUNOHISTOCELLTY SEE INCREASES 383 00:16:14,280 --> 00:16:16,200 IN CADHERIN 1 GENE EXPRESSION, 384 00:16:16,200 --> 00:16:21,520 WHICH IS E-CADHERIN, AND WE SEE 385 00:16:21,520 --> 00:16:22,080 E-CADHERIN HISTOCHEMICALLY 386 00:16:22,080 --> 00:16:23,720 AROUND THE ECM AND GRANULOMAS AS 387 00:16:23,720 --> 00:16:30,160 WELL AS THINGS THAT LOOK LIKE 388 00:16:30,160 --> 00:16:33,920 TERTIARY LYMPHOID STRUCTURES, 389 00:16:33,920 --> 00:16:35,200 CO-LOCATING WITH CDPOSITIVE 390 00:16:35,200 --> 00:16:35,440 CELLS. 391 00:16:35,440 --> 00:16:38,480 THERE'S RECENT PAPER THAT SHOWED 392 00:16:38,480 --> 00:16:41,880 THAT ACTIVATION OF BETA-CATENIN 393 00:16:41,880 --> 00:16:45,800 WAS LINKED WITH MUSCLE ATROPHY, 394 00:16:45,800 --> 00:16:48,640 MUSCLE WASTING. 395 00:16:48,640 --> 00:16:50,280 E-CADHERIN ENGAGEMENT PREVENTS 396 00:16:50,280 --> 00:16:51,000 TRANSLOCATION TO THE NUCLEUS, 397 00:16:51,000 --> 00:16:53,160 THIS MIGHT BE A WAY FOR OUR 398 00:16:53,160 --> 00:16:55,120 BODIES TO HELP TRY AND SAVE SOME 399 00:16:55,120 --> 00:16:57,400 MUSCLE DURING AN INJURY AND 400 00:16:57,400 --> 00:16:58,400 PREVENT ATROPHY. 401 00:16:58,400 --> 00:17:02,560 SO WE LOOKED AT CD103, WHAT 402 00:17:02,560 --> 00:17:05,880 ABOUT CCR 1? 403 00:17:05,880 --> 00:17:08,120 BINDS THE CHEMOKINE IN MICE AND 404 00:17:08,120 --> 00:17:11,760 HUMANS, HUMANS ALSO HAVE XCL2, 405 00:17:11,760 --> 00:17:12,960 ASSOCIATED WITH CHEMOTAXIS OF 406 00:17:12,960 --> 00:17:15,040 THE CELLS TO TUMORS. 407 00:17:15,040 --> 00:17:18,400 WHEN WE LOOK AT PROTEIN AND GENE 408 00:17:18,400 --> 00:17:19,760 EXPRESSION, TOP ROW LOCAL 409 00:17:19,760 --> 00:17:23,240 TISSUE, WE SEE IN THAT MUSCLE WE 410 00:17:23,240 --> 00:17:24,880 CAN UPREGULATION WITH THE ECM 411 00:17:24,880 --> 00:17:28,040 TREATMENT AND WE ALSO SEE THAT 412 00:17:28,040 --> 00:17:30,040 WITH THE TISSUE, THE WHOLE 413 00:17:30,040 --> 00:17:31,440 MUSCLE HOMOGENIZED AT ONCE, IF 414 00:17:31,440 --> 00:17:33,280 WE ISOLATE SINGLE CELLS WE SEE 415 00:17:33,280 --> 00:17:35,160 THAT UPREGULATION. 416 00:17:35,160 --> 00:17:37,360 THE LACK OF UPREGULATION AND 417 00:17:37,360 --> 00:17:39,200 POLYETHYLENE TREATMENT IS A 418 00:17:39,200 --> 00:17:42,920 POSSIBLE EXPLANATION WHY WE 419 00:17:42,920 --> 00:17:44,360 AREN'T SEEING SEEING CDC1s, 420 00:17:44,360 --> 00:17:45,800 NOT GETTING THE CHEMOKINE SIGNAL 421 00:17:45,800 --> 00:17:46,840 TO GET RECRUITED IN. 422 00:17:46,840 --> 00:17:48,400 IF WE LOOK IN THE BLOOD 423 00:17:48,400 --> 00:17:50,600 TREMENDOUS WE SEE PEAK EARLY ON, 424 00:17:50,600 --> 00:17:52,680 ASSOCIATED WITH THE RECRUITMENT 425 00:17:52,680 --> 00:17:56,520 OF THESE CELLS, AND THEN 426 00:17:56,520 --> 00:17:57,800 INTERESTINGLY RAG KNOCKOUT MODEL 427 00:17:57,800 --> 00:18:00,840 WITHOUT T CELLS OR B CELLS WE 428 00:18:00,840 --> 00:18:04,840 DON'T SEE CHANGES WITH XCL-1, 429 00:18:04,840 --> 00:18:08,120 SUGGESTING IT'S SOURCED FROM 430 00:18:08,120 --> 00:18:09,440 INDEPENDENT SUBTYPE, PRODUCED BY 431 00:18:09,440 --> 00:18:10,680 T CELLS AND NK CELLS. 432 00:18:10,680 --> 00:18:12,840 T CELLS ARE RAG DEPENDENT SO WE 433 00:18:12,840 --> 00:18:15,760 LEANED TOWARD IT BEING THESE NK 434 00:18:15,760 --> 00:18:16,200 CELLS. 435 00:18:16,200 --> 00:18:18,840 AS MENTIONED THERE ARE 436 00:18:18,840 --> 00:18:23,760 LITERATURE REPORTS OF NC CELL 437 00:18:23,760 --> 00:18:26,080 MEDIATED CDC-1 TUMORS, AND SO WE 438 00:18:26,080 --> 00:18:27,960 SORTED OUT LYMPHOCYTES AND BY WE 439 00:18:27,960 --> 00:18:31,080 I MEAN 5:00 IN THE MORNING, 440 00:18:31,080 --> 00:18:33,200 STARTING AN EXPERIMENT WE, I WAS 441 00:18:33,200 --> 00:18:34,280 THERE, I PROMISE. 442 00:18:34,280 --> 00:18:35,600 AND HAD SOME GREAT TEAM WORK TO 443 00:18:35,600 --> 00:18:37,240 GET THIS DATA. 444 00:18:37,240 --> 00:18:40,720 AGAIN, ANOTHER GOOD DATA DAY SO 445 00:18:40,720 --> 00:18:51,120 WE SAW 300-FOLD HIGHER XCL-1, SO 446 00:18:51,120 --> 00:18:52,560 IT APPEARS NATURAL KILLER CELLS 447 00:18:52,560 --> 00:18:55,400 ARE PRODUCING THIS. 448 00:18:55,400 --> 00:18:58,360 A POST BACK STIMULATED NC CELLS 449 00:18:58,360 --> 00:18:59,280 IN VITRO WITH FRAGMENT 450 00:18:59,280 --> 00:19:01,840 EXTRACELLULAR MATRIX AND DAMAGE 451 00:19:01,840 --> 00:19:05,920 ASSOCIATED MOLECULAR PATTERN, 452 00:19:05,920 --> 00:19:08,000 LOW MOLECULAR WEIGHT HYALURONIC 453 00:19:08,000 --> 00:19:08,320 ACID. 454 00:19:08,320 --> 00:19:13,280 AND WE DID SEE WITHIN 24 HOURS A 455 00:19:13,280 --> 00:19:15,120 MODEST UPREGULATION IN THESE NC 456 00:19:15,120 --> 00:19:18,280 CELLS, IT DOES APPEAR NATURAL 457 00:19:18,280 --> 00:19:21,480 KILLER CELLS ARE GETTING TISSUE 458 00:19:21,480 --> 00:19:26,280 DAMAGE SIGNALS INDUCING TO 459 00:19:26,280 --> 00:19:27,360 SECRETE THIS. 460 00:19:27,360 --> 00:19:28,560 DO THEY REGULATE TISSUE 461 00:19:28,560 --> 00:19:29,760 REGENERATION AFTER INJURY? 462 00:19:29,760 --> 00:19:31,360 WE SEE ALL OF THESE CHANGES TO 463 00:19:31,360 --> 00:19:33,040 IMMUNE CELLS, WHAT ABOUT THE 464 00:19:33,040 --> 00:19:34,600 TISSUE ITSELF? 465 00:19:34,600 --> 00:19:39,640 SO IN OUR WILD TYPE MICE 466 00:19:39,640 --> 00:19:45,760 NORMALLY YOU GET INQUINNAL FAT 467 00:19:45,760 --> 00:19:47,720 BED MIGRATION, DENSE 468 00:19:47,720 --> 00:19:49,560 INFILTRATION AROUND THE 469 00:19:49,560 --> 00:19:53,960 PARTICLES, INTO THE ECM SCAFFOLD 470 00:19:53,960 --> 00:20:01,600 BUT NO DENSE FIBROTIC CAB CYLE. 471 00:20:01,600 --> 00:20:04,880 WITH BATF3 WE SEE DEPOSITION 472 00:20:04,880 --> 00:20:06,800 SUGGESTING ISSUES WITH 473 00:20:06,800 --> 00:20:08,000 PHAGOCYTOSIS, MORE WHITE FAT 474 00:20:08,000 --> 00:20:10,040 WHERE IT SHOULDN'T BE, AND WE 475 00:20:10,040 --> 00:20:12,680 ALSO SAW MORE DISTAL FROM THE 476 00:20:12,680 --> 00:20:15,720 INJURY SIDE MUSCLE FIBER 477 00:20:15,720 --> 00:20:17,520 NECROSIS AND CALCIFICATION, 478 00:20:17,520 --> 00:20:19,480 CALCIUM IS STAINED BLACK ON THE 479 00:20:19,480 --> 00:20:20,440 RIGHT. 480 00:20:20,440 --> 00:20:25,040 WE FOLLOWED THIS WITH PCR, HAD 481 00:20:25,040 --> 00:20:26,920 UPREGULATION ASSOCIATED WITH 482 00:20:26,920 --> 00:20:30,400 WHITE FAT, ALSO INCREASE IN 483 00:20:30,400 --> 00:20:32,080 ACVR1, A RECEPTOR THAT'S MUTATED 484 00:20:32,080 --> 00:20:34,480 IN A HUMAN DISEASE ASSOCIATED 485 00:20:34,480 --> 00:20:37,400 WITH SEVERE POSTTRAUMATIC SOFT 486 00:20:37,400 --> 00:20:38,760 TISSUE OSSIFICATION. 487 00:20:38,760 --> 00:20:40,040 INTERESTINGLY, WE ACTUALLY WOUND 488 00:20:40,040 --> 00:20:45,120 UP SEEING INCREASES IN MUSCLE 489 00:20:45,120 --> 00:20:46,320 REGENERATION MARKERS AS WELL. 490 00:20:46,320 --> 00:20:48,400 THIS WAS ODD FOR US BUT IF WE 491 00:20:48,400 --> 00:20:50,360 LOOK BACK AT OUR HISTOLOGY AND 492 00:20:50,360 --> 00:20:52,880 LOOK BACK AT THAT MUSCLE MOST 493 00:20:52,880 --> 00:20:56,360 MUSCLE FIBERS DO APPEAR TO HAVE 494 00:20:56,360 --> 00:20:57,880 AN INCREASE IN MUSCLE FIBER 495 00:20:57,880 --> 00:20:58,920 CROSS-SECTIONAL AREA. 496 00:20:58,920 --> 00:21:01,600 SO WE MIGHT BE SEEING DISRUPTION 497 00:21:01,600 --> 00:21:04,760 IN THE REGULATION OF OVERALL 498 00:21:04,760 --> 00:21:06,640 TISSUE DEVELOPMENT AFTER INJURY. 499 00:21:06,640 --> 00:21:09,800 IN GENERAL WE HAVE THIS CIRCUIT 500 00:21:09,800 --> 00:21:11,560 HERE THAT'S SURROUNDED BY 501 00:21:11,560 --> 00:21:19,120 DENDRITIC CELLS WORKING WITH 502 00:21:19,120 --> 00:21:20,440 MACROPHAGES, EOSINOPHILS AND 503 00:21:20,440 --> 00:21:22,160 TREGS ALL WORKING TOGETHER TO 504 00:21:22,160 --> 00:21:23,240 ULTIMATELY DETERMINE WHAT THE 505 00:21:23,240 --> 00:21:25,880 OUTCOME IS IN TERMS OF TISSUE 506 00:21:25,880 --> 00:21:26,840 DEVELOPMENT. 507 00:21:26,840 --> 00:21:29,840 NOW, WHAT'S OUR CURRENT 508 00:21:29,840 --> 00:21:34,760 HYPOTHESIS REGARDING THIS KIND 509 00:21:34,760 --> 00:21:35,520 OF SELF-REGULATION TO 510 00:21:35,520 --> 00:21:35,840 AUTOIMMUNITY? 511 00:21:35,840 --> 00:21:37,160 IF WE THINK ABOUT IT AFTER 512 00:21:37,160 --> 00:21:40,920 INJURY WE HAVE THE RELEASE OF 513 00:21:40,920 --> 00:21:42,360 DAMAGE ASSOCIATED MOLECULAR 514 00:21:42,360 --> 00:21:43,440 PATTERNS, SELF ANTIGEN PRESENT 515 00:21:43,440 --> 00:21:48,720 OUTSIDE HOMEO TASTE IS SO -- 516 00:21:48,720 --> 00:21:49,680 HOMEOSTASIS. 517 00:21:49,680 --> 00:21:52,760 WE GET THIS RELEASE OF XCL-1 518 00:21:52,760 --> 00:21:55,600 FROM THE INJURY SITE, 519 00:21:55,600 --> 00:21:57,480 RECRUITMENT OF CDC1s, WOULD BE 520 00:21:57,480 --> 00:22:00,680 ABLE TO CROSS-PRESENT THIS 521 00:22:00,680 --> 00:22:03,240 ANTIGEN TO TREGS, AND GET 522 00:22:03,240 --> 00:22:03,680 IMMUNOREGULATION AND 523 00:22:03,680 --> 00:22:06,120 SELF-TOLERANCE IN THE MORE 524 00:22:06,120 --> 00:22:07,200 REGULATORY ENVIRONMENT 525 00:22:07,200 --> 00:22:08,000 CHARACTERIZED BY IL-10 526 00:22:08,000 --> 00:22:09,520 EXPRESSION. 527 00:22:09,520 --> 00:22:10,840 IF YOU THINK ABOUT CANCER, 528 00:22:10,840 --> 00:22:13,480 CANCER IS USING PATHWAY TO 529 00:22:13,480 --> 00:22:16,520 PROTECT ITSELF DURING DISRUPTED 530 00:22:16,520 --> 00:22:17,960 HOMEOSTASIS. 531 00:22:17,960 --> 00:22:19,040 SO THIS TISSUE INJURY AS MANY 532 00:22:19,040 --> 00:22:20,600 HAVE SAID THAT CANCER IS THIS 533 00:22:20,600 --> 00:22:22,640 WOUND THAT NEVER HEALS SO WE CAN 534 00:22:22,640 --> 00:22:24,520 THINK OF TISSUE INJURY ACTUALLY 535 00:22:24,520 --> 00:22:26,600 CREATING A PATHWAY THAT COULD 536 00:22:26,600 --> 00:22:28,880 ALLOW CANCER TO SURVIVE. 537 00:22:28,880 --> 00:22:30,960 AND SO SUBJECT TO CHANGE, BUT 538 00:22:30,960 --> 00:22:33,360 THAT'S OUR CURRENT THINKING. 539 00:22:33,360 --> 00:22:36,200 AND SO JUST SOME QUICK 540 00:22:36,200 --> 00:22:40,080 ACKNOWLEDGMENTS, THOSE 541 00:22:40,080 --> 00:22:42,840 HIGHLIGHTED IN TEAL HELPED OUT 542 00:22:42,840 --> 00:22:44,960 WITH THIS PROJECT A LOT, THANK 543 00:22:44,960 --> 00:22:48,560 YOU, AS WELL AS OUR LEADERSHIP 544 00:22:48,560 --> 00:22:50,760 FROM NIBIB, RICHARD AND BRUCE 545 00:22:50,760 --> 00:22:54,280 HAVE BEEN SUPPORTIVE OF OUR LAB, 546 00:22:54,280 --> 00:22:55,280 AND THEN OUR ADMINISTRATIVE 547 00:22:55,280 --> 00:22:56,800 OFFICERS THAT HAVE HELPED OUT AS 548 00:22:56,800 --> 00:22:59,080 WELL AS MY OWN MENTORS AND 549 00:22:59,080 --> 00:23:01,280 ADVOCATES OUTSIDE NIH THAT HAVE 550 00:23:01,280 --> 00:23:03,880 BEEN HELPFUL IN MY CAREER 551 00:23:03,880 --> 00:23:05,440 JOURNEY SO I MASSIVELY THANK 552 00:23:05,440 --> 00:23:05,640 THEM. 553 00:23:05,640 --> 00:23:07,280 I'M HAPPY TO TAKE QUESTIONS NOW 554 00:23:07,280 --> 00:23:10,160 OR VIA E-MAIL IF YOU'D LIKE. 555 00:23:10,160 --> 00:23:11,920 THANK YOU VERY MUCH. 556 00:23:11,920 --> 00:23:19,840 [APPLAUSE] 557 00:23:19,840 --> 00:23:20,960 >> TOO MUCH IMMUNOLOGY? 558 00:23:20,960 --> 00:23:21,720 >> THANKS FOR THIS. 559 00:23:21,720 --> 00:23:27,840 I TRY TO STAY AWAY FROM 560 00:23:27,840 --> 00:23:28,160 IMMUNOLOGY. 561 00:23:28,160 --> 00:23:32,240 A QUICK QUESTION. 562 00:23:32,240 --> 00:23:37,600 SOME OF THE DIFFERENT -- AT 563 00:23:37,600 --> 00:23:39,080 IMMUNOLOGY TALKS, MOUSE 564 00:23:39,080 --> 00:23:42,160 IMMUNOLOGY, A QUESTION ABOUT 565 00:23:42,160 --> 00:23:44,040 HUMAN, COULD YOU LET US KNOW 566 00:23:44,040 --> 00:23:47,320 WHICH CELL TYPES HAVE BEEN 567 00:23:47,320 --> 00:23:50,080 IDENTIFIED IN HUMANS, EVEN 568 00:23:50,080 --> 00:23:52,040 NATURAL MATERIAL, CLINICAL 569 00:23:52,040 --> 00:23:53,040 TRIALS INTO THE CLINIC? 570 00:23:53,040 --> 00:23:54,480 >>YEAH, THE INTERESTING THING 571 00:23:54,480 --> 00:23:56,440 ABOUT THIS CASCADE IS THAT IT IS 572 00:23:56,440 --> 00:23:57,640 CONSERVED IN HUMANS. 573 00:23:57,640 --> 00:23:58,320 THAT WAS SOMETHING THAT WAS 574 00:23:58,320 --> 00:23:58,840 INTERESTING FOR US. 575 00:23:58,840 --> 00:24:01,600 ONE OF THE REASONS I DIDN'T 576 00:24:01,600 --> 00:24:05,480 KNOW, IT WAS A MOUSE MARKER, I 577 00:24:05,480 --> 00:24:09,040 DIDN'T EVER FOLLOW INTO ITS 578 00:24:09,040 --> 00:24:09,640 FUNCTION. 579 00:24:09,640 --> 00:24:17,560 CD103 IN HUMANS, E-CADHERIN IN 580 00:24:17,560 --> 00:24:21,080 HUMANS, XCR 1, XCL 1, WE'RE 581 00:24:21,080 --> 00:24:22,480 PLANNING ON FOLLOWING IN HUMANS 582 00:24:22,480 --> 00:24:24,040 TO CROSS-CHECK AND HAVE 583 00:24:24,040 --> 00:24:26,560 INDEPENDENT RESEARCH SCHOLAR IN 584 00:24:26,560 --> 00:24:28,720 THE GROUP WHO IS COLLABORATING 585 00:24:28,720 --> 00:24:31,160 WITH NICOLE MORGAN'S GROUP ON 586 00:24:31,160 --> 00:24:31,760 DEVELOPING MORE MICROFLUIDIC 587 00:24:31,760 --> 00:24:32,880 MODELS AND SO HER LAB IS GOING 588 00:24:32,880 --> 00:24:35,280 TO BE FOCUSED ON KIND OF GETTING 589 00:24:35,280 --> 00:24:37,480 SOME MORE OF THOSE HUMAN MODELS 590 00:24:37,480 --> 00:24:38,440 ESTABLISHED THAT WE CAN START 591 00:24:38,440 --> 00:24:40,000 PLAYING AROUND WITH. 592 00:24:40,000 --> 00:24:42,200 SO WE'RE EXCITED BECAUSE IT DOES 593 00:24:42,200 --> 00:24:43,720 APPEAR TO BE -- IT'S THE SAME 594 00:24:43,720 --> 00:24:46,320 PATHWAYS IN HUMANS, AND WE'RE 595 00:24:46,320 --> 00:24:48,640 CURRENTLY WORKING ON GETTING A 596 00:24:48,640 --> 00:24:51,280 COURIER SERVICE TO ACTUALLY GET 597 00:24:51,280 --> 00:24:52,920 THE HUMAN SAMPLES, SO BITS OF 598 00:24:52,920 --> 00:24:54,680 PAPERWORK LEFT BUT WE'RE 599 00:24:54,680 --> 00:25:01,440 CHECKING ON THAT. 600 00:25:01,440 --> 00:25:04,720 >>YOU TALK IN TERMS OF CANCER 601 00:25:04,720 --> 00:25:06,880 RESEARCH, ARE YOU ALSO LOOKING 602 00:25:06,880 --> 00:25:11,920 AT AUTOIMMUNE DISEASES IN 603 00:25:11,920 --> 00:25:12,240 GENERAL? 604 00:25:12,240 --> 00:25:12,560 (INAUDIBLE). 605 00:25:12,560 --> 00:25:14,760 >> I KNOW NOTHING ABOUT CANCER. 606 00:25:14,760 --> 00:25:16,800 WE FOCUS IN ON THIS TRAUMATIC 607 00:25:16,800 --> 00:25:17,840 INJURY SPACE. 608 00:25:17,840 --> 00:25:19,360 IT'S INTERESTING TO SEE CANCER 609 00:25:19,360 --> 00:25:21,440 MANIPULATES SOME OF THOSE 610 00:25:21,440 --> 00:25:22,120 PATHWAYS. 611 00:25:22,120 --> 00:25:25,480 AND SO I THINK THAT THESE SORT 612 00:25:25,480 --> 00:25:26,920 OF EXPERIMENTS, THIS SORT OF 613 00:25:26,920 --> 00:25:29,960 INVESTIGATION, COULD HELP LEND 614 00:25:29,960 --> 00:25:32,040 SOME INSIGHT INTO EVEN 615 00:25:32,040 --> 00:25:32,800 AUTOIMMUNE DISEASES THAT ARE NOT 616 00:25:32,800 --> 00:25:33,720 NECESSARILY SOMETHING YOU THINK 617 00:25:33,720 --> 00:25:36,960 ARE ASSOCIATED WITH AN INJURY. 618 00:25:36,960 --> 00:25:39,480 WE KNOW THAT PERSISTENT 619 00:25:39,480 --> 00:25:41,800 INFLAMMATION SUCH AS COVID-19 620 00:25:41,800 --> 00:25:42,800 CAN TRIGGER AUTOIMMUNE-LIKE 621 00:25:42,800 --> 00:25:43,080 CONDITIONS. 622 00:25:43,080 --> 00:25:47,040 A RECENT PAPER CAME OUT I THINK 623 00:25:47,040 --> 00:25:49,320 YESTERDAY, DAY BEFORE, DOUBLE 624 00:25:49,320 --> 00:25:52,720 INCREASED RISK FOR TYPE 1 625 00:25:52,720 --> 00:25:56,440 DIABETES IN PATIENTS WITH 626 00:25:56,440 --> 00:25:58,280 COVID-19, INFLAMMATION CAN 627 00:25:58,280 --> 00:26:00,600 INDUCE THOSE, THOUGH IT'S FRAMED 628 00:26:00,600 --> 00:26:03,120 WITHIN TRAUMATIC INJURY AND 629 00:26:03,120 --> 00:26:04,200 TISSUE RECONSTRUCTION COULD 630 00:26:04,200 --> 00:26:08,920 HOPEFULLY OFFER INSIGHT INTO 631 00:26:08,920 --> 00:26:09,960 OTHER AREAS. 632 00:26:09,960 --> 00:26:12,040 >> ONE MORE THING. 633 00:26:12,040 --> 00:26:18,360 (INAUDIBLE). 634 00:26:18,360 --> 00:26:19,960 >> WE HAVEN'T MEASURED 635 00:26:19,960 --> 00:26:22,040 BIOMECHANICAL PROPERTIES OF IT. 636 00:26:22,040 --> 00:26:25,880 WHAT WE DID WAS TOOK THOSE DENSE 637 00:26:25,880 --> 00:26:28,600 PLANTAR SHEETS, GOING TO HAVE 638 00:26:28,600 --> 00:26:30,320 THE SAME FORMULATION, BUT 639 00:26:30,320 --> 00:26:33,720 PROVIDED IN A DIFFERENT WAY, 640 00:26:33,720 --> 00:26:34,840 VERSUS THE PARTICULATE, AND SO 641 00:26:34,840 --> 00:26:36,040 WE DID LOOK AT THAT. 642 00:26:36,040 --> 00:26:37,760 WE DIDN'T SEE A DIFFERENCE, 643 00:26:37,760 --> 00:26:39,200 THOUGH MECHANICS ARE REALLY 644 00:26:39,200 --> 00:26:41,160 IMPORTANT IN IMMUNE ACTIVATION 645 00:26:41,160 --> 00:26:42,920 AND POLARIZATION SO PREVIOUS 646 00:26:42,920 --> 00:26:44,360 STUDIES ASSOCIATED WITH MORE 647 00:26:44,360 --> 00:26:46,200 SYNTHETIC HYDROGELS WHERE WE 648 00:26:46,200 --> 00:26:47,520 MODIFIED STIFFNESS OF THAT 649 00:26:47,520 --> 00:26:49,840 MATERIAL, YOU CAN SEE 650 00:26:49,840 --> 00:26:51,040 STIFFNESS-DEPENDENT CHANGE IN 651 00:26:51,040 --> 00:26:51,600 RESPONSE. 652 00:26:51,600 --> 00:26:54,080 AND SO WE CERTAINLY HAVE DONE A 653 00:26:54,080 --> 00:26:57,240 FEW OF THOSE EXPERIMENTS WITH 654 00:26:57,240 --> 00:27:00,080 SYNTHETIC HYDROGELS AND MANY 655 00:27:00,080 --> 00:27:02,640 OTHERS OUT THERE THAT ARE 656 00:27:02,640 --> 00:27:03,600 BIOMATERIAL MECHANICS AND HAVE 657 00:27:03,600 --> 00:27:06,920 DONE WONDERFUL WORK ON THAT. 658 00:27:06,920 --> 00:27:09,400 659 00:27:09,400 --> 00:27:15,640 >> (INAUDIBLE). 660 00:27:15,640 --> 00:27:19,240 >> YES, WE LOVE TO BE ABLE TO 661 00:27:19,240 --> 00:27:22,000 DEVELOP MORE SYNTHETIC KIND OF 662 00:27:22,000 --> 00:27:23,320 CONTROLLED HYDROGELS, WHERE IT'S 663 00:27:23,320 --> 00:27:26,040 X, Y AND Z AND WHAT'S IN EVERY 664 00:27:26,040 --> 00:27:27,880 SINGLE BATCH SO HOPEFULLY TRYING 665 00:27:27,880 --> 00:27:30,200 TO LEARN THE BASICS OF HOW 666 00:27:30,200 --> 00:27:32,040 SOMETHING LIKE AN EXTRACELLULAR 667 00:27:32,040 --> 00:27:33,920 MATRIX SCAFFOLD WHICH HELPS WITH 668 00:27:33,920 --> 00:27:35,880 THIS IMMUNE RECRUITMENT AND MORE 669 00:27:35,880 --> 00:27:36,760 POSITIVE WOUNDING OUTCOMES, SO 670 00:27:36,760 --> 00:27:38,840 IF WE CAN LEARN FROM THAT TO 671 00:27:38,840 --> 00:27:41,320 THEN APPLY TO THE DESIGN HONING 672 00:27:41,320 --> 00:27:44,640 IN ON WHAT ARE THE GOOD THINGS 673 00:27:44,640 --> 00:27:45,320 ABOUT THAT MATERIAL, SO, FOR 674 00:27:45,320 --> 00:27:48,000 EXAMPLE, ONE OF THE THINGS WE 675 00:27:48,000 --> 00:27:49,960 SEE, THIS REALLY HIGH EOSINOPHIL 676 00:27:49,960 --> 00:27:52,800 RECRUITMENT EARLY ON, DO WE WANT 677 00:27:52,800 --> 00:27:54,240 THAT MEAN EOSINOPHILS? WE DON'T 678 00:27:54,240 --> 00:27:55,040 KNOW YET. 679 00:27:55,040 --> 00:27:56,440 RAVI IS WORKING ON THAT 680 00:27:56,440 --> 00:27:56,720 CURRENTLY. 681 00:27:56,720 --> 00:27:58,320 SO WE'RE TRYING TO TEASE OUT THE 682 00:27:58,320 --> 00:27:59,960 GOOD AND BAD OF THOSE MATERIALS 683 00:27:59,960 --> 00:28:03,680 SO WE CAN GO AHEAD AND TAKE 684 00:28:03,680 --> 00:28:05,320 THOSE PUZZLE PIECES AND PUT THEM 685 00:28:05,320 --> 00:28:07,880 INTO SOMETHING THAT'S CONTROLLED 686 00:28:07,880 --> 00:28:09,200 AND HAS MINIMAL BATCH-TO-BATCH 687 00:28:09,200 --> 00:28:11,560 VARIABILITY. 688 00:28:11,560 --> 00:28:14,920 689 00:28:14,920 --> 00:28:15,440 THANK YOU SO MUCH. 690 00:28:15,440 --> 00:28:17,800 [APPLAUSE] 691 00:28:17,800 --> 00:28:23,960 692 00:28:23,960 --> 00:28:26,920 >> IF ANYONE HAS QUESTIONS -- 693 00:28:26,920 --> 00:28:34,920 (INAUDIBLE). 694 00:28:34,920 --> 00:28:36,440 >> THANK YOU. 695 00:28:36,440 --> 00:28:39,600 NEXT SPEAKER IS DR. MATTHEW 696 00:28:39,600 --> 00:28:41,280 WOLFE, STADTMAN INVESTIGATOR, 697 00:28:41,280 --> 00:28:43,320 HEAD OF CANCER BIOMATERIALS 698 00:28:43,320 --> 00:28:45,640 ENGINEERING SECTION FROM NCI. 699 00:28:45,640 --> 00:28:49,320 LAB IS INVESTIGATING THE 700 00:28:49,320 --> 00:28:50,680 IMMUNOMODULATORY BIOMATERIALS TO 701 00:28:50,680 --> 00:28:54,080 USE IN NEXT GENERATION CANCER 702 00:28:54,080 --> 00:28:54,760 IMMUNOLOGY. 703 00:28:54,760 --> 00:28:55,120 THANK YOU. 704 00:28:55,120 --> 00:28:55,640 >> ALL RIGHT. 705 00:28:55,640 --> 00:28:59,880 THANK YOU VERY MUCH FOR INVITING 706 00:28:59,880 --> 00:29:04,000 ME, FOR THAT INTRODUCTION. 707 00:29:04,000 --> 00:29:04,360 OKAY. 708 00:29:04,360 --> 00:29:12,800 SO, DID THE SLIDES COME UP? 709 00:29:12,800 --> 00:29:14,080 I'M FROM THE NCI, FREDERICK, 710 00:29:14,080 --> 00:29:14,840 MARYLAND. 711 00:29:14,840 --> 00:29:18,480 I'M ALSO A STADTMAN 712 00:29:18,480 --> 00:29:19,600 INVESTIGATOR, STARTED AUGUST 713 00:29:19,600 --> 00:29:20,040 2020. 714 00:29:20,040 --> 00:29:24,960 AND I ALSO USE BIOMATERIALS AND 715 00:29:24,960 --> 00:29:26,600 ALSO USE EXTRACELLULAR MATRIX 716 00:29:26,600 --> 00:29:27,600 FROM DECELLULARRIZED TISSUES. 717 00:29:27,600 --> 00:29:33,960 WHAT I'M GOING TO TALK ABOUT 718 00:29:33,960 --> 00:29:35,920 TODAY -- OH -- IS USING THESE 719 00:29:35,920 --> 00:29:37,920 MATERIALS IN A VERY DIFFERENT 720 00:29:37,920 --> 00:29:39,960 WAY. 721 00:29:39,960 --> 00:29:46,000 722 00:29:46,000 --> 00:29:53,120 723 00:29:53,120 --> 00:29:58,120 LET'S SEE IF THIS WORKS. 724 00:29:58,120 --> 00:30:00,920 ANYWAYS ... 725 00:30:00,920 --> 00:30:01,440 >> (INAUDIBLE). 726 00:30:01,440 --> 00:30:04,200 >> OH, I HOPE NOT. 727 00:30:04,200 --> 00:30:08,600 THAT'S MISERABLE. 728 00:30:08,600 --> 00:30:12,520 729 00:30:12,520 --> 00:30:14,520 THERE WE GO. 730 00:30:14,520 --> 00:30:17,000 THANK YOU. 731 00:30:17,000 --> 00:30:24,560 732 00:30:24,560 --> 00:30:25,960 ALL RIGHT. 733 00:30:25,960 --> 00:30:30,120 SO LIKE I MENTIONED, WE ARE 734 00:30:30,120 --> 00:30:31,000 BIOMATERIALS LAB. 735 00:30:31,000 --> 00:30:32,680 AND WHAT YOU KNOW IN ADDITION TO 736 00:30:32,680 --> 00:30:35,600 SOME THINGS THAT CAITLIN POINTED 737 00:30:35,600 --> 00:30:37,760 OUT, MATERIALS PLAY AN IMPORTANT 738 00:30:37,760 --> 00:30:43,040 ROLE IN CANCER. 739 00:30:43,040 --> 00:30:44,600 SCAFFOLD, POROUS CELLS CAN BE 740 00:30:44,600 --> 00:30:46,840 USED TO DELIVER DRUGS, IN A 741 00:30:46,840 --> 00:30:47,280 CONTROLLED FASHION. 742 00:30:47,280 --> 00:30:51,920 THEY CAN BE USED IN MODELING 743 00:30:51,920 --> 00:30:53,320 DIFFERENT TISSUE SYSTEMS IN DRUG 744 00:30:53,320 --> 00:30:53,640 DEVELOPMENT. 745 00:30:53,640 --> 00:31:00,040 AND THEY CAN BE USED HOW CAITLIN 746 00:31:00,040 --> 00:31:03,080 DESCRIBED IN A TISSUE 747 00:31:03,080 --> 00:31:05,920 REPAIRATIVE CONTEXT, AND THEY 748 00:31:05,920 --> 00:31:07,680 CAN BE IMMUNE MODULATORY, THAT'S 749 00:31:07,680 --> 00:31:10,080 DEFINITELY TRUE IN THE PREVIOUS 750 00:31:10,080 --> 00:31:10,720 TALK. 751 00:31:10,720 --> 00:31:13,680 THE DRUG DELIVERY ASPECT AND 752 00:31:13,680 --> 00:31:15,400 IMMUNE MODULATORY ASPECT WILL BE 753 00:31:15,400 --> 00:31:25,920 THE THEMES I HIGHLIGHT TODAY. 754 00:31:25,920 --> 00:31:26,880 DECELLULARIZATION TURNS A 755 00:31:26,880 --> 00:31:28,880 TRANSPLANT INTO A MEDICAL 756 00:31:28,880 --> 00:31:29,480 DEVICE. 757 00:31:29,480 --> 00:31:34,920 REMOVE THE CELLS, PREVENT ACUTE 758 00:31:34,920 --> 00:31:35,520 REJECTION. 759 00:31:35,520 --> 00:31:36,640 AND WE VERIFY THAT A NUMBER OF 760 00:31:36,640 --> 00:31:40,480 WAYS, AS DO OTHERS IN THE FIELD. 761 00:31:40,480 --> 00:31:43,320 LET ME SEE IF I CAN GET A 762 00:31:43,320 --> 00:31:44,960 POINTER UP HERE. 763 00:31:44,960 --> 00:31:47,920 LOOKING AT REMOVAL OF ALL THESE 764 00:31:47,920 --> 00:31:50,880 NUCLEI AND SMOOTH MUSCLE ACTIN, 765 00:31:50,880 --> 00:31:52,960 SMALL INTESTINE, SAME THAT 766 00:31:52,960 --> 00:31:56,600 CAITLIN MENTIONED, WE VERIFY DNA 767 00:31:56,600 --> 00:31:57,360 REMOVAL, QUANTITATIVELY, AND 768 00:31:57,360 --> 00:31:59,880 MAKING SURE THAT WHATEVER IS 769 00:31:59,880 --> 00:32:03,680 LEFT DNA-WISE IS EXTREMELY 770 00:32:03,680 --> 00:32:03,880 SMALL. 771 00:32:03,880 --> 00:32:05,680 AND IT'S NOT JUST INTESTINE, 772 00:32:05,680 --> 00:32:06,720 THOUGH THAT'S WHAT I'M GOING TO 773 00:32:06,720 --> 00:32:07,840 FOCUS ON TODAY. 774 00:32:07,840 --> 00:32:11,120 CAN YOU DO THIS WITH VIRTUALLY 775 00:32:11,120 --> 00:32:12,880 ANY SOLID TISSUE FROM ANY ANIMAL 776 00:32:12,880 --> 00:32:15,160 OR HUMAN SOURCE, IT'S NOT JUST 777 00:32:15,160 --> 00:32:16,040 ACADEMICS USING THIS. 778 00:32:16,040 --> 00:32:18,560 THEY ARE USED ALL THE TIME IN 779 00:32:18,560 --> 00:32:21,400 THE CLINIC, SO BOTH HUMAN AND 780 00:32:21,400 --> 00:32:24,040 ANIMAL TISSUES ARE USED AS 781 00:32:24,040 --> 00:32:26,680 SOURCE MATERIALS FOR THESE ECM 782 00:32:26,680 --> 00:32:28,440 SCAFFOLDED FOR TISSUE REPAIR IN 783 00:32:28,440 --> 00:32:30,400 THE CLINIC, THESE ARE JUST 784 00:32:30,400 --> 00:32:32,840 HANDFUL OF EXAMPLES CURRENTLY ON 785 00:32:32,840 --> 00:32:33,560 THE MARKET. 786 00:32:33,560 --> 00:32:38,400 AND WHAT WE ARE DOING IS WE'RE 787 00:32:38,400 --> 00:32:40,720 LEVERAGING MATERIALS AND 788 00:32:40,720 --> 00:32:42,000 VACCINES, ESPECIALLY CANCER 789 00:32:42,000 --> 00:32:42,920 VACCINE. 790 00:32:42,920 --> 00:32:46,280 AND SO IN THIS APPROACH, YOU 791 00:32:46,280 --> 00:32:48,040 TAKE BIOMATERIAL SCAFFOLD AND 792 00:32:48,040 --> 00:32:52,400 LOAD IT OR INFUSE IT WITH CANCER 793 00:32:52,400 --> 00:32:54,480 VACCINE COMPONENTS, ANTIGENS, 794 00:32:54,480 --> 00:32:54,840 ADJUVANTS. 795 00:32:54,840 --> 00:32:56,440 AND WITH ANY DRUG DELIVERY 796 00:32:56,440 --> 00:32:58,480 APPROACH LIKE THIS THERE ARE 797 00:32:58,480 --> 00:33:00,040 CERTAIN ADVANTAGES WHEN YOU 798 00:33:00,040 --> 00:33:01,160 IMPLANT THAT IN VIVO. 799 00:33:01,160 --> 00:33:03,200 ONE IS THAT YOU PROLONG EXPOSURE 800 00:33:03,200 --> 00:33:07,400 OF THE BODY OF THE IMMUNE SYSTEM 801 00:33:07,400 --> 00:33:08,880 TO VACCINE EXPONENTS RATHER THAN 802 00:33:08,880 --> 00:33:10,640 IT GETTING FLUSHED AWAY, WASHED 803 00:33:10,640 --> 00:33:12,720 AWAY IMMEDIATELY. 804 00:33:12,720 --> 00:33:15,240 THE MATERIAL ITSELF IS ALSO 805 00:33:15,240 --> 00:33:16,120 IMMUNOSTIMULATORY, THAT'S ONE OF 806 00:33:16,120 --> 00:33:17,960 THOSE FEATURES WE TRY TO 807 00:33:17,960 --> 00:33:19,760 CAPITALIZE ON. 808 00:33:19,760 --> 00:33:21,160 IT'S ATTRACTING LEUKOCYTES TO 809 00:33:21,160 --> 00:33:23,240 WHERE THAT CANCER VACCINE 810 00:33:23,240 --> 00:33:23,680 EXISTS. 811 00:33:23,680 --> 00:33:25,360 YOU KNOW, TO THE THERAPEUTIC YOU 812 00:33:25,360 --> 00:33:27,600 WANT IT TO RESPOND TO. 813 00:33:27,600 --> 00:33:29,040 ULTIMATELY THIS PRODUCED A 814 00:33:29,040 --> 00:33:30,880 POLARIZED IMMUNE RESPONSE THAT 815 00:33:30,880 --> 00:33:32,200 IF DEVELOPED IN APPROPRIATE WAY 816 00:33:32,200 --> 00:33:36,680 CAN TRAVEL TO OTHER SITES TO 817 00:33:36,680 --> 00:33:38,000 ATTACK CANCER CELLS. 818 00:33:38,000 --> 00:33:40,280 AND TO DATE THE MAJORITY OF 819 00:33:40,280 --> 00:33:42,920 MATERIALS USED IN THIS SCAFFOLD 820 00:33:42,920 --> 00:33:44,560 DELIVERY APPROACH HAVE BEEN 821 00:33:44,560 --> 00:33:47,880 THOSE FOREIGN BODY REACTION TYPE 822 00:33:47,880 --> 00:33:52,440 OF INFLAMMATORY RESPONSES, LIKE 823 00:33:52,440 --> 00:33:54,000 PLGA, OR ALGINATE. 824 00:33:54,000 --> 00:33:57,920 AND AS CAITLIN MENTIONED, ECM 825 00:33:57,920 --> 00:33:59,480 SCAFFOLD ARE CERTAINLY IMMUNE 826 00:33:59,480 --> 00:34:00,000 STIMULATORY. 827 00:34:00,000 --> 00:34:03,200 AND DO MANY OF THESE SAME 828 00:34:03,200 --> 00:34:03,400 THINGS. 829 00:34:03,400 --> 00:34:04,720 BUT POLARIZE DIFFERENT IMMUNE 830 00:34:04,720 --> 00:34:05,320 RESPONSE. 831 00:34:05,320 --> 00:34:08,040 REALLY THIS IS REHASHING WHAT 832 00:34:08,040 --> 00:34:09,960 WAS MENTIONED ALREADY, A LOT OF 833 00:34:09,960 --> 00:34:14,120 TYPE 2 BIASED IMMUNE FEATURES, 834 00:34:14,120 --> 00:34:21,360 CD206 POSITIVE MACROPHAGE, AND 835 00:34:21,360 --> 00:34:24,840 LOW BUT NOT INSIGNIFICANT CD8 T 836 00:34:24,840 --> 00:34:25,120 CELLS. 837 00:34:25,120 --> 00:34:28,560 SO SINCE THESE MATERIALS ARE 838 00:34:28,560 --> 00:34:31,400 USED EVERY DAY TO REPAIR TISSUE 839 00:34:31,400 --> 00:34:33,480 FOLLOWING TUMOR RESECTION CAN WE 840 00:34:33,480 --> 00:34:35,440 LEVERAGE THIS SCAFFOLD IMPLANT 841 00:34:35,440 --> 00:34:38,520 IN A CANCER VACCINE 842 00:34:38,520 --> 00:34:38,840 IMMUNOTHERAPY? 843 00:34:38,840 --> 00:34:41,040 AND A QUESTION THAT COMES UP ON 844 00:34:41,040 --> 00:34:43,520 THE WAY IS THIS IDEA OF TYPE 2 845 00:34:43,520 --> 00:34:48,080 IMMUNITY IN CANCER. 846 00:34:48,080 --> 00:34:51,880 REMEMBER THOSE IL-4-DRIVEN 847 00:34:51,880 --> 00:34:53,080 CD4-DRIVEN MACROPHAGE 848 00:34:53,080 --> 00:34:53,760 206-EXPRESSING MACROPHAGE 849 00:34:53,760 --> 00:34:54,600 RESPONSES, CLASSICALLY THOUGHT 850 00:34:54,600 --> 00:34:57,560 OF AS IMMUNE SUPPRESSIVE IN THE 851 00:34:57,560 --> 00:34:59,200 CONTEXT OF CANCER. 852 00:34:59,200 --> 00:35:00,760 BUT I WOULD CHALLENGE THAT 853 00:35:00,760 --> 00:35:02,360 SIMPLY BECAUSE IT DOESN'T MATCH 854 00:35:02,360 --> 00:35:04,560 WHAT WE FIND IN CLINICAL OR 855 00:35:04,560 --> 00:35:07,000 PRE-CLINICAL DATA. 856 00:35:07,000 --> 00:35:10,240 IN FACT, SUCH ECM MATERIALS 857 00:35:10,240 --> 00:35:12,760 SOMETIMES CALLED BIOLOGICS ARE 858 00:35:12,760 --> 00:35:14,440 ACTUALLY PREFERABLY USED IN 859 00:35:14,440 --> 00:35:15,000 CONTAMINATED FIELDS WHETHER 860 00:35:15,000 --> 00:35:16,040 THERE'S A HIGH RISK OF 861 00:35:16,040 --> 00:35:17,080 INFECTION. 862 00:35:17,080 --> 00:35:21,080 NOT SOMETHING WHETHER YOU WANT 863 00:35:21,080 --> 00:35:22,160 BLANKET IMMUNE SUPPRESSION. 864 00:35:22,160 --> 00:35:25,320 I POSIT IT'S A DIFFERENT TYPE OF 865 00:35:25,320 --> 00:35:26,960 INFLAMMATION THAT IS NOT BLANKET 866 00:35:26,960 --> 00:35:29,840 IMMUNE SUPPRESSION AND WE CAN 867 00:35:29,840 --> 00:35:32,320 USE THIS TYPE OF LEUKOCYTE 868 00:35:32,320 --> 00:35:34,960 ATTRACTION TO SYNERGIZE WITH 869 00:35:34,960 --> 00:35:36,720 LOCALIZED CANCER VACCINE. 870 00:35:36,720 --> 00:35:38,560 WE WANTED TO REALLY ANSWER THE 871 00:35:38,560 --> 00:35:44,040 QUESTION OF WHETHER THESE ECM 872 00:35:44,040 --> 00:35:54,600 SCAFFOLDS CAN MODULATE CYTOTOXIC 873 00:35:58,880 --> 00:35:59,120 IMMUNITY. 874 00:35:59,120 --> 00:36:00,720 WE WANTED TO STUDY INDUCING 875 00:36:00,720 --> 00:36:10,840 EFFECT ON OF -- FROM CANCER 876 00:36:10,840 --> 00:36:11,440 VACCINE. 877 00:36:11,440 --> 00:36:14,480 LOADING A FORM OF ECM WITH TUMOR 878 00:36:14,480 --> 00:36:18,000 ANTIGEN AND WITH A 879 00:36:18,000 --> 00:36:18,680 CYTOTOXIC-INDUCING ADJUVANT, 880 00:36:18,680 --> 00:36:20,400 HIGHLIGHTED HERE BECAUSE WE 881 00:36:20,400 --> 00:36:22,360 DON'T GET THAT TYPE OF RESPONSE 882 00:36:22,360 --> 00:36:24,640 INHERENTLY FROM ECM SO WE ADD 883 00:36:24,640 --> 00:36:25,960 ADDITIONAL COMPOUNDS TO INDUCE 884 00:36:25,960 --> 00:36:26,160 THAT. 885 00:36:26,160 --> 00:36:28,480 CAN WE USE IT TO TREAT 886 00:36:28,480 --> 00:36:29,440 ESTABLISHED TUMORS? 887 00:36:29,440 --> 00:36:30,440 WE'LL USE A MOUSE MODEL. 888 00:36:30,440 --> 00:36:33,520 ALONG THE WAY WE CAN ASK SEVERAL 889 00:36:33,520 --> 00:36:34,240 QUESTIONS. 890 00:36:34,240 --> 00:36:35,840 SO, IN ADDITION TO FUNDAMENTAL 891 00:36:35,840 --> 00:36:38,640 ONE DOES THIS ECM SCAFFOLD 892 00:36:38,640 --> 00:36:39,560 MICROENVIRONMENT AFFECT 893 00:36:39,560 --> 00:36:41,160 CYTOTOXIC C TELL FUNCTION? 894 00:36:41,160 --> 00:36:42,600 HOW DOES THE IMMUNE ADJUVANT 895 00:36:42,600 --> 00:36:47,520 THAT WE'RE ADDING AFFECT HOST 896 00:36:47,520 --> 00:36:50,920 RESPONSE AND HOW DOES THIS 897 00:36:50,920 --> 00:36:53,000 COLLABORATE TO CREATE A 898 00:36:53,000 --> 00:36:56,480 PRODUCTIVE ANTITUMOR RESPONSE 899 00:36:56,480 --> 00:36:57,880 WITH CANCER VACCINE. 900 00:36:57,880 --> 00:37:08,040 WE'RE ALSO USING A MUCOSA ECM 901 00:37:08,040 --> 00:37:08,520 PARTICULATE, CLINICALLY 902 00:37:08,520 --> 00:37:09,200 RELEVANT, WELL CHARACTERIZED, A 903 00:37:09,200 --> 00:37:10,080 GOOD PLACE TO START. 904 00:37:10,080 --> 00:37:13,640 SO WE TAKE THAT ECM, GRIND INTO 905 00:37:13,640 --> 00:37:15,200 PARTICLES, THAT ARE ABOUT 50 906 00:37:15,200 --> 00:37:16,960 MICRONS IN SIZE, IMPORTANTLY ARE 907 00:37:16,960 --> 00:37:18,720 INJECTABLE AND SOMETHING WE CAN 908 00:37:18,720 --> 00:37:21,680 MIX WITH OUR CANCER VACCINE OF 909 00:37:21,680 --> 00:37:24,040 TUMOR ANTIGEN AND IMMUNE 910 00:37:24,040 --> 00:37:24,320 ADJUVANTS. 911 00:37:24,320 --> 00:37:27,880 AND AS A QUICK OVERVIEW, ANY 912 00:37:27,880 --> 00:37:29,560 GOOD CANCER VACCINE HAS AT 913 00:37:29,560 --> 00:37:30,840 MINIMUM TWO COMPONENTS. 914 00:37:30,840 --> 00:37:33,160 THAT TUMOR ANTIGEN THAT PROVIDES 915 00:37:33,160 --> 00:37:34,360 THE FINGERPRINT THAT IDENTIFIES 916 00:37:34,360 --> 00:37:37,240 CANCER CELLS FROM YOUR NORMAL 917 00:37:37,240 --> 00:37:40,040 CELLS AND THEN IMMUNE ADJUVANT 918 00:37:40,040 --> 00:37:42,920 TO ATTRACT AND POLARIZE 919 00:37:42,920 --> 00:37:45,120 RESPONSES AGAINST THAT ANTIGEN. 920 00:37:45,120 --> 00:37:48,360 AND SO WHEN YOU DELIVER A CANCER 921 00:37:48,360 --> 00:37:49,760 VACCINE YOU'RE INITIATING THE 922 00:37:49,760 --> 00:37:50,760 CASCADE OF IMMUNE INTERACTIONS, 923 00:37:50,760 --> 00:37:52,640 THIS IS JUST ONE BRANCH OF 924 00:37:52,640 --> 00:37:53,960 WHAT'S GOING ON, BUT ULTIMATELY 925 00:37:53,960 --> 00:37:56,440 WHAT I'M GOING TO BE FOCUSING ON 926 00:37:56,440 --> 00:38:01,960 TODAY IS ABILITY TO INDUCE 927 00:38:01,960 --> 00:38:03,600 ANTIGEN-SPECIFIC TUMOR ANTIGEN 928 00:38:03,600 --> 00:38:06,240 SPECIFIC CD8 CELLS TO LYSE TUMOR 929 00:38:06,240 --> 00:38:06,640 CELLS. 930 00:38:06,640 --> 00:38:10,160 SO THE FIRST QUESTION, WHAT'S AN 931 00:38:10,160 --> 00:38:11,600 APPROPRIATE ADJUVANT TO INDUCE 932 00:38:11,600 --> 00:38:13,760 CYTOTOXIC RESPONSE WHEN 933 00:38:13,760 --> 00:38:14,960 DELIVERED WITH ECM SCAFFOLD? 934 00:38:14,960 --> 00:38:17,080 I WON'T GO THROUGH ALL THESE IN 935 00:38:17,080 --> 00:38:20,440 A LOT OF DETAIL OTHER THAN TO 936 00:38:20,440 --> 00:38:23,760 SAY THEY EACH STIMULATE A 937 00:38:23,760 --> 00:38:25,160 DIFFERENT PATHWAY AND DIFFERENT 938 00:38:25,160 --> 00:38:33,400 RESPONSE ON THE WAY TO 939 00:38:33,400 --> 00:38:34,720 INITIATING CONTRIBUTING 940 00:38:34,720 --> 00:38:42,040 CYTOTOXIC RESPONSE. . 941 00:38:42,040 --> 00:38:43,680 WE CAST A WIDE NET BECAUSE NO 942 00:38:43,680 --> 00:38:50,760 ONE HAS DONE THIS BEFORE WITH 943 00:38:50,760 --> 00:38:52,440 ECM SCAFFOLD MATERIAL. 944 00:38:52,440 --> 00:38:53,360 THIS IS BASELINE 945 00:38:53,360 --> 00:38:54,600 CHARACTERIZATION TO STUDY EFFECT 946 00:38:54,600 --> 00:38:56,480 OF WHAT ADJUVANTS WITH DOING TO 947 00:38:56,480 --> 00:38:59,320 THE SCAFFOLD HOST RESPONSE 948 00:38:59,320 --> 00:39:00,040 ITSELF. 949 00:39:00,040 --> 00:39:02,960 SO HERE IS THE SIS IMPLANT IN A 950 00:39:02,960 --> 00:39:04,240 SUBCUTANEOUS ENVIRONMENT, NO 951 00:39:04,240 --> 00:39:08,480 INJURY HERE, JUST A SUB Q 952 00:39:08,480 --> 00:39:10,160 INJECTION IN A MOUSE. 953 00:39:10,160 --> 00:39:13,000 YOU GET IMMUNE ACTIVATION AND 954 00:39:13,000 --> 00:39:14,200 INFILTRATION ON ITS OWN, YOU CAN 955 00:39:14,200 --> 00:39:17,280 SEE A BUNCH OF CELLS, MAJORITY 956 00:39:17,280 --> 00:39:19,360 OF IMMUNE CELLS INFILTRATING THE 957 00:39:19,360 --> 00:39:22,200 SAS IMPLANT ON ITS OWN. 958 00:39:22,200 --> 00:39:24,280 IN CONTRAST, SOMETHING LIKE 959 00:39:24,280 --> 00:39:27,040 MPLA, A POTENT IMMUNE ADJUVANT, 960 00:39:27,040 --> 00:39:30,360 CREATES A MUCH MORE ROBUST 961 00:39:30,360 --> 00:39:31,800 CELLULAR INFILTRATE SO EVEN 962 00:39:31,800 --> 00:39:34,000 AFTER 14 DAYS, WHICH IS WHEN 963 00:39:34,000 --> 00:39:35,760 THESE RESPONSES SHOULD BE DYING 964 00:39:35,760 --> 00:39:37,480 DOWN, WE SEE SOMETHING THAT'S 965 00:39:37,480 --> 00:39:42,200 VERY DENSELY PACKED WITH CELLS, 966 00:39:42,200 --> 00:39:43,720 ALMOST INFECTION LIKE, QUANTIFY 967 00:39:43,720 --> 00:39:45,360 FROM HISTOLOGY ACROSS A TIME 968 00:39:45,360 --> 00:39:49,200 COURSE, AND INDEED THAT'S WHAT 969 00:39:49,200 --> 00:39:59,720 WE SEE, BOTH HAVE THIS AFTER ONE 970 00:39:59,720 --> 00:40:02,760 DAY, CD8 DECREASED CURIOUSLY 971 00:40:02,760 --> 00:40:03,760 THAT INITIAL INFILTRATION, 972 00:40:03,760 --> 00:40:06,280 EVERYTHING CAUGHT UP BY DAY 7, 973 00:40:06,280 --> 00:40:10,320 AND ONLY MPLA ARE THE CHRONIC 974 00:40:10,320 --> 00:40:10,920 INFLAMMATORY ENVIRONMENT AFTER 975 00:40:10,920 --> 00:40:13,240 14 DAYS. 976 00:40:13,240 --> 00:40:15,240 THE ADJUVANTS ARE CHANGING AND 977 00:40:15,240 --> 00:40:19,280 BOTH INFILTRATION NUMBER AND 978 00:40:19,280 --> 00:40:21,680 KINETICS OF THAT RESPONSE. 979 00:40:21,680 --> 00:40:23,320 THERE WE GO. 980 00:40:23,320 --> 00:40:25,400 SO WE ALSO CHARACTERIZE WHAT 981 00:40:25,400 --> 00:40:27,520 THOSE IMMUNE CELLS ARE. 982 00:40:27,520 --> 00:40:31,680 SO THERE ARE CERTAIN IMMUNE 983 00:40:31,680 --> 00:40:34,960 HALLMARKS ASSOCIATED WITH UCM 984 00:40:34,960 --> 00:40:35,560 IMPLANTATION, SO CD206 985 00:40:35,560 --> 00:40:37,240 MACROPHAGES BEING A BIG PART OF 986 00:40:37,240 --> 00:40:37,560 THAT. 987 00:40:37,560 --> 00:40:41,280 SO THOSE ARE INDUCED HIGHLY WHEN 988 00:40:41,280 --> 00:40:43,480 ECM IS IMPLANTED ALONE. 989 00:40:43,480 --> 00:40:44,800 AND SOMETIMES CALLED AN M2 990 00:40:44,800 --> 00:40:48,040 MARKER IF YOU FOLLOW THAT 991 00:40:48,040 --> 00:40:48,280 PARADIGM. 992 00:40:48,280 --> 00:40:50,800 WE ALSO LOOK AT CD8 6, WHICH IS 993 00:40:50,800 --> 00:40:52,440 IMPORTANT IN VACCINE RESPONSES 994 00:40:52,440 --> 00:40:58,000 AND T CELL PRIMING. 995 00:40:58,000 --> 00:41:03,480 AND WHAT WE FIND HERE ONLY THE 996 00:41:03,480 --> 00:41:07,760 CD8 GROUP INDUCE APPRECIATABLE 997 00:41:07,760 --> 00:41:09,160 UPREGULATION OF THAT CD86 THAT'S 998 00:41:09,160 --> 00:41:17,720 SO IMPORTANT FOR T CELL PRIMING 999 00:41:17,720 --> 00:41:18,800 AND ACTIVATION. 1000 00:41:18,800 --> 00:41:20,360 AND REALLY ONLY MPLA WAS 1001 00:41:20,360 --> 00:41:25,400 TRENDING TO DECREASE IN HIGH 206 1002 00:41:25,400 --> 00:41:26,040 REGULATION. 1003 00:41:26,040 --> 00:41:29,280 WE ALSO LOOK AT LYMPH NODES AS 1004 00:41:29,280 --> 00:41:30,840 SENTINELS OF WHAT'S GOING ON IN 1005 00:41:30,840 --> 00:41:32,560 THE REGIONAL ENVIRONMENT, NOT 1006 00:41:32,560 --> 00:41:35,640 JUST LOCALLY WITHIN THE 1007 00:41:35,640 --> 00:41:36,080 SCAFFOLD. 1008 00:41:36,080 --> 00:41:39,040 IL-4 BEING ANOTHER HALLMARK OF 1009 00:41:39,040 --> 00:41:41,000 ECM RESPONSE, SIS MATERIAL ALONE 1010 00:41:41,000 --> 00:41:44,960 INDUCES A LARGE UPREGULATION OF 1011 00:41:44,960 --> 00:41:46,240 IL-4, SURPRISINGLY ADJUVANTS 1012 00:41:46,240 --> 00:41:48,800 DIDN'T DIMINISH THAT EVEN THOUGH 1013 00:41:48,800 --> 00:41:50,960 THEY ARE CYTOTOXIC INDUCING, IN 1014 00:41:50,960 --> 00:41:58,480 SOME CASES ENHANCED IT IN FACT. 1015 00:41:58,480 --> 00:41:59,480 OKAY. 1016 00:41:59,480 --> 00:42:02,240 AND SO AFTER CHARACTERIZING 1017 00:42:02,240 --> 00:42:05,080 WHAT'S GOING ON LOCALLY AND 1018 00:42:05,080 --> 00:42:06,520 REGIONALLY THE GOAL IS TO 1019 00:42:06,520 --> 00:42:09,560 ULTIMATELY LOOK FOR FUNCTION SO 1020 00:42:09,560 --> 00:42:11,320 ARE WE GETTING CYTOTOXIC T 1021 00:42:11,320 --> 00:42:14,240 CELLS, ANTIGEN SPECIFIC T CELLS 1022 00:42:14,240 --> 00:42:14,800 FOLLOWING THIS APPROACH? 1023 00:42:14,800 --> 00:42:17,360 SO, TO DO THAT WE USE WHAT'S 1024 00:42:17,360 --> 00:42:21,840 KNOWN AS IN VIVO CTL ASSAY, A 1025 00:42:21,840 --> 00:42:24,680 QUANTITATIVE METRIC OF CD8 T 1026 00:42:24,680 --> 00:42:26,080 CELL KILLING, ANTIGEN SPECIFIC 1027 00:42:26,080 --> 00:42:26,320 FASHION. 1028 00:42:26,320 --> 00:42:34,200 AND SO WE TOOK OUR ECM, SIS ECM, 1029 00:42:34,200 --> 00:42:36,960 COMBINED WITH A MODEL ANTIGEN 1030 00:42:36,960 --> 00:42:47,440 USEFUL FOR TRACKING ANTIGEN 1031 00:42:49,880 --> 00:42:51,960 SPECIFICITY AND ADJUVANT. 1032 00:42:51,960 --> 00:42:53,760 EVEN WITH SALINE ALONE, THE 1033 00:42:53,760 --> 00:42:57,120 SOLUBLE DELIVER WRITE OF OCA AND 1034 00:42:57,120 --> 00:42:59,760 CDA WERE MAXING OUT ASSAY BUT 1035 00:42:59,760 --> 00:43:01,920 NOT LOSING ANYTHING BY 1036 00:43:01,920 --> 00:43:02,920 DELIVERING ARE OUR ECM. 1037 00:43:02,920 --> 00:43:05,000 AND THAT'S NOT THE CASE FOR SOME 1038 00:43:05,000 --> 00:43:09,080 OF THESE OTHER ADJUVANTS WHERE 1039 00:43:09,080 --> 00:43:10,880 MPLA BECOMES VARIABLE IN A 1040 00:43:10,880 --> 00:43:14,840 DECREASE. 1041 00:43:14,840 --> 00:43:17,240 WE HAVE A COMPATIBLE ADJUVANT 1042 00:43:17,240 --> 00:43:23,360 FOR INDUCING SKY TOE CYTOTOXIC 1043 00:43:23,360 --> 00:43:23,760 IMMUNITY. 1044 00:43:23,760 --> 00:43:25,960 ANOTHER IS TO PROLONG DELIVERY 1045 00:43:25,960 --> 00:43:26,920 OF THESE COMPOUNDS, PREVENT THEM 1046 00:43:26,920 --> 00:43:28,360 FROM GOING AWAY TOO QUICKLY AND 1047 00:43:28,360 --> 00:43:29,440 GETTING THEM TO STICK AROUND. 1048 00:43:29,440 --> 00:43:31,280 WE WANT TO SEE IF WE'RE GETTING 1049 00:43:31,280 --> 00:43:34,640 A BENEFIT FROM USING AN ECM 1050 00:43:34,640 --> 00:43:35,800 SCAFFOLD, WHEN DELIVERED IN 1051 00:43:35,800 --> 00:43:43,320 VIVO, FOR THAT WE TOOK THAT 1052 00:43:43,320 --> 00:43:46,600 OVALBUMIN MODEL, CONJUGATED TO 1053 00:43:46,600 --> 00:43:48,040 NEAR-INFRARED DYE FOR 1054 00:43:48,040 --> 00:43:49,560 FLUORESCENCE IMAGE, TRACKED MICE 1055 00:43:49,560 --> 00:43:51,320 OVER TIME, QUANTIFIED THE 1056 00:43:51,320 --> 00:43:53,880 RESPONSE, AND WHAT WE FOUND WAS 1057 00:43:53,880 --> 00:43:58,800 INDEED WITH RESPECT TO JUST 1058 00:43:58,800 --> 00:44:03,520 DELIVER THE OVA WITH SALINE 1059 00:44:03,520 --> 00:44:04,600 SUBCUTANEOUSLY, WE'RE ABLE TO 1060 00:44:04,600 --> 00:44:10,440 GET MORE SUSTAINED RELEASE AT 1061 00:44:10,440 --> 00:44:11,640 INJECTION SITE WITH SIS. 1062 00:44:11,640 --> 00:44:13,680 ALUM WAS OLD AS POSITIVE 1063 00:44:13,680 --> 00:44:16,200 CONTROL, AN OLD VACCINE ADJUVANT 1064 00:44:16,200 --> 00:44:17,200 THAT'S GOOD AT BINDING PROTEIN 1065 00:44:17,200 --> 00:44:20,200 AND KEEPING IT AT THE INJECTION 1066 00:44:20,200 --> 00:44:20,400 SITE. 1067 00:44:20,400 --> 00:44:23,480 WHAT I WANT TO POSIT HERE, TOO 1068 00:44:23,480 --> 00:44:24,680 GOOD IN THIS INSTANCE, APPEARS 1069 00:44:24,680 --> 00:44:27,840 TO BE LOCKED UP AT THAT 1070 00:44:27,840 --> 00:44:28,520 INJECTION SITE, RATHER THAN 1071 00:44:28,520 --> 00:44:30,160 BEING TAKEN UP BY THE IMMUNE 1072 00:44:30,160 --> 00:44:34,840 SYSTEM SO THAT IT CAN THEN BE 1073 00:44:34,840 --> 00:44:38,680 USED FOR ANTIGEN PROCESSING. 1074 00:44:38,680 --> 00:44:40,760 WHEREAS WE'RE GETTING EXTENDED 1075 00:44:40,760 --> 00:44:42,840 RELEASE WITH SIS OVER TIME LINE 1076 00:44:42,840 --> 00:44:44,840 THAT'S RELEVANT FOR VACCINATION 1077 00:44:44,840 --> 00:44:46,240 APPROACH. 1078 00:44:46,240 --> 00:44:50,600 SO WHERE IS THAT OVA GOING? 1079 00:44:50,600 --> 00:44:53,320 WELL, WE FIND THAT IT'S BEING 1080 00:44:53,320 --> 00:44:59,920 INTERNIZED BY MYELOID CELLS, 1081 00:44:59,920 --> 00:45:04,720 TAKING THAT FLUORESCENT OVA, CAN 1082 00:45:04,720 --> 00:45:10,720 VACCINATE, COLLECT MYELOID CELLS 1083 00:45:10,720 --> 00:45:11,480 EXPRESSING CD11B, AND WHAT WE 1084 00:45:11,480 --> 00:45:15,560 FIND IS THAT WE DO INDEED FIND 1085 00:45:15,560 --> 00:45:16,560 THOSE OVA-POSITIVE MYELOID CELLS 1086 00:45:16,560 --> 00:45:18,280 THAT HAVE EATEN ANTIGEN AND MADE 1087 00:45:18,280 --> 00:45:20,520 THEIR WAY TO THE LYMPH NODE IN 1088 00:45:20,520 --> 00:45:23,080 ORDER TO PRESENT IT TO T CELLS, 1089 00:45:23,080 --> 00:45:25,280 AND THIS INCREASES SUBSTANTIALLY 1090 00:45:25,280 --> 00:45:29,000 WHEN CDA IS USED AS ADJUVANT. 1091 00:45:29,000 --> 00:45:30,520 THE CONTRALATERAL LYMPH NODE, 1092 00:45:30,520 --> 00:45:35,360 JUST AS OUR NEGATIVE CONTROL. 1093 00:45:35,360 --> 00:45:35,760 OKAY. 1094 00:45:35,760 --> 00:45:38,360 SO, THE PUNCHLINE IS DOES THIS 1095 00:45:38,360 --> 00:45:40,840 WORK IN TREATING ESTABLISHED 1096 00:45:40,840 --> 00:45:41,800 TUMORS? 1097 00:45:41,800 --> 00:45:47,400 SO WHEN USING OUR VACCINE, OUR 1098 00:45:47,400 --> 00:45:48,840 SIS AS ECM SCAFFOLD COMPONENT, 1099 00:45:48,840 --> 00:45:55,280 CDA ADJUVANT WITH MOST EFFICACY 1100 00:45:55,280 --> 00:46:01,440 IN CTLA ASSAY INDUCING ANTIGEN 1101 00:46:01,440 --> 00:46:04,280 SPECIFIC CD8 RESPONSE AND 1102 00:46:04,280 --> 00:46:05,480 OVALBUMIN, THESE ARE CELLS THAT 1103 00:46:05,480 --> 00:46:09,080 FORM TUMORS WHEN THEY ARE 1104 00:46:09,080 --> 00:46:11,160 INJECTED SUBCUTANEOUSLY AND 1105 00:46:11,160 --> 00:46:12,160 EXPRESS THAT OVA ANTIGEN, 1106 00:46:12,160 --> 00:46:13,480 SOMETHING WE CAN VACCINATE 1107 00:46:13,480 --> 00:46:15,760 AGAINST IN OUR MODEL SYSTEM 1108 00:46:15,760 --> 00:46:16,000 HERE. 1109 00:46:16,000 --> 00:46:19,400 SO, ONCE TUMORS ARE ESTABLISHED 1110 00:46:19,400 --> 00:46:20,440 WE THEN THERAPEUTICALLY 1111 00:46:20,440 --> 00:46:24,720 VACCINATE ONCE AND AGAIN SEVEN 1112 00:46:24,720 --> 00:46:25,160 DAYS LATER. 1113 00:46:25,160 --> 00:46:30,560 AND SO WHAT YOU CAN SEE HERE IS 1114 00:46:30,560 --> 00:46:31,680 THAT UNTREATED ANIMALS TUMORS 1115 00:46:31,680 --> 00:46:35,720 GROW VERY RAPIDLY AS EXPECTED. 1116 00:46:35,720 --> 00:46:37,720 WHEN THE VACCINE IS PROVIDED IN 1117 00:46:37,720 --> 00:46:44,800 A SOLUBLE FORM JUST CDA AND OVA 1118 00:46:44,800 --> 00:46:48,000 SUSPENDED IN SALINE, PARTIAL 1119 00:46:48,000 --> 00:46:51,600 RESPONSE, ALL TUMORS EVENTUALLY 1120 00:46:51,600 --> 00:46:53,800 GROW AND REQUIRE SACRIFICE. 1121 00:46:53,800 --> 00:46:55,560 HOWEVER, WHEN DELIVERING WITH 1122 00:46:55,560 --> 00:46:58,640 SIS ECM SCAFFOLD WE GET GREATER 1123 00:46:58,640 --> 00:47:02,880 AN 50% RESPONSES SO TUMORS 1124 00:47:02,880 --> 00:47:03,960 REGRESS UPON TREATMENT, AND 1125 00:47:03,960 --> 00:47:06,480 ABOUT HALF OF THEM OR A LITTLE 1126 00:47:06,480 --> 00:47:14,040 MORE THAN HALF DON'T COME BACK. 1127 00:47:14,040 --> 00:47:15,440 AND SO THIS MANIFESTS AS 1128 00:47:15,440 --> 00:47:16,120 SURVIVAL BENEFIT COMPARED TO 1129 00:47:16,120 --> 00:47:16,760 OTHER GROUPS. 1130 00:47:16,760 --> 00:47:17,960 WHEN WE LOOK AT THESE ANIMALS 1131 00:47:17,960 --> 00:47:20,600 THE ONES THAT HAVE FULLY 1132 00:47:20,600 --> 00:47:22,360 REJECTED THEIR TUMORS UPON 1133 00:47:22,360 --> 00:47:23,960 VACCINATION, WE LET THEM GO FOR 1134 00:47:23,960 --> 00:47:25,720 A FEW MONTHS, FOR A COUPLE 1135 00:47:25,720 --> 00:47:27,240 MONTHS HERE TO MAKE SURE THEY 1136 00:47:27,240 --> 00:47:31,640 DON'T COME BACK AND PERFORM A 1137 00:47:31,640 --> 00:47:32,120 RECHALLENGE EXPERIMENT. 1138 00:47:32,120 --> 00:47:33,600 WHERE WE INJECT THE VERY SAME 1139 00:47:33,600 --> 00:47:35,040 CANCER LINE ON THE OTHER SIDE. 1140 00:47:35,040 --> 00:47:41,040 SO THAT WILL TELL US WHETHER WE 1141 00:47:41,040 --> 00:47:42,040 HAVE SYSTEMIC PROTECTION, 1142 00:47:42,040 --> 00:47:44,880 WHETHER THE MOUSE HAS BEEN 1143 00:47:44,880 --> 00:47:45,640 SUCCESSFULLY VACCINATED WITH 1144 00:47:45,640 --> 00:47:46,600 MEMORY RESPONSE. 1145 00:47:46,600 --> 00:47:50,440 INDEED THAT'S WHAT WE FIND HERE. 1146 00:47:50,440 --> 00:47:52,640 SO, AFTER THAT RECHALLENGE, OUR 1147 00:47:52,640 --> 00:47:54,920 NAIVE AGE MATCHED CONTROLS 1148 00:47:54,920 --> 00:47:56,160 RAPIDLY FORMED TUMORS AS 1149 00:47:56,160 --> 00:47:57,880 EXPECTED AND NOT A SINGLE MOUSE 1150 00:47:57,880 --> 00:48:00,000 WAS LOST FROM THE VACCINATED 1151 00:48:00,000 --> 00:48:01,720 GROUP SO WE HAVE IMMUNE MEMORY 1152 00:48:01,720 --> 00:48:04,800 AND PROTECTION IN THE LONG TERM 1153 00:48:04,800 --> 00:48:06,200 AND WE'RE CUTTING IT OFF HERE 1154 00:48:06,200 --> 00:48:07,960 BUT THEY WENT FOR A WHILE AFTER 1155 00:48:07,960 --> 00:48:10,360 THIS AS WELL. 1156 00:48:10,360 --> 00:48:14,200 AND FINALLY WE WANTED TO DO A 1157 00:48:14,200 --> 00:48:15,400 LITTLE VALIDATION THAT WHAT WE 1158 00:48:15,400 --> 00:48:19,120 THINK IS HAPPENING IS ACTUALLY 1159 00:48:19,120 --> 00:48:22,120 HAPPENING. 1160 00:48:22,120 --> 00:48:26,000 SO FROM OUR CTL ASSAY WE KNEW 1161 00:48:26,000 --> 00:48:28,120 THIS GENERATED CYTOTOXIC T CELLS 1162 00:48:28,120 --> 00:48:29,880 SPECIFIC FOR OVA BUT ARE THEY 1163 00:48:29,880 --> 00:48:31,600 RESPONSIBLE FOR TUMOR 1164 00:48:31,600 --> 00:48:32,000 REGRESSION? 1165 00:48:32,000 --> 00:48:32,920 THERE'S STILL STEPS INVOLVED 1166 00:48:32,920 --> 00:48:35,240 WHERE THE T CELLS HAVE TO 1167 00:48:35,240 --> 00:48:36,280 INFILTRATE AND BECOME EFFECTIVE, 1168 00:48:36,280 --> 00:48:39,600 IN THAT ENVIRONMENT AS WELL. 1169 00:48:39,600 --> 00:48:42,000 SO WE CONFIRMED THAT CD8 T CELLS 1170 00:48:42,000 --> 00:48:45,200 WERE DOING THIS USING CELL 1171 00:48:45,200 --> 00:48:48,040 DEPLETION EXPERIMENT, MONOCLONAL 1172 00:48:48,040 --> 00:48:51,400 ANTIBODIES, WE CAN DEPLETE 1173 00:48:51,400 --> 00:48:53,600 DIFFERENT IMMUNE SUBTYPES FROM 1174 00:48:53,600 --> 00:48:57,680 MICE BEARING EG 7 TUMORS, AND BY 1175 00:48:57,680 --> 00:49:01,800 APPLYING ANTI-CD8 TREATMENT WE 1176 00:49:01,800 --> 00:49:02,640 EFFECTIVELY REDUCE CIRCULATING 1177 00:49:02,640 --> 00:49:06,080 CYTOTOXIC CD8 T CELLS TO ZERO. 1178 00:49:06,080 --> 00:49:09,480 AND SO IN THIS EXPERIMENT, AS 1179 00:49:09,480 --> 00:49:11,040 EXPECTED, THE UNTREATED MICE 1180 00:49:11,040 --> 00:49:12,640 AGAIN GREW VERY RAPIDLY, THOSE 1181 00:49:12,640 --> 00:49:17,680 THAT WERE VACCINATED BUT GOT THE 1182 00:49:17,680 --> 00:49:19,880 CONTROL ANTIBODY AGAIN 50% OR 1183 00:49:19,880 --> 00:49:23,920 OVER 50% RESPONSE RATES, AS 1184 00:49:23,920 --> 00:49:24,800 BEFORE. 1185 00:49:24,800 --> 00:49:26,680 AND WHEN DEPLETING CD8 T CELLS 1186 00:49:26,680 --> 00:49:28,880 THAT EFFECT IS COMPLETELY LOST. 1187 00:49:28,880 --> 00:49:32,040 SO, THESE CD8 T CELLS ARE 1188 00:49:32,040 --> 00:49:33,720 ABSOLUTELY CRUCIAL AND NECESSARY 1189 00:49:33,720 --> 00:49:37,760 FOR FUNCTIONAL TUMOR REGRESSION 1190 00:49:37,760 --> 00:49:40,840 SO WE ARE INDUCING THIS SYSTEMIC 1191 00:49:40,840 --> 00:49:44,000 ANTITUMOR RESPONSE IN A 1192 00:49:44,000 --> 00:49:46,040 CD8-DEPENDENT FASHION FOLLOWING 1193 00:49:46,040 --> 00:49:47,920 THIS SIS ECM VACCINE. 1194 00:49:47,920 --> 00:49:52,040 SO, I'LL STOP THERE AND OFFER 1195 00:49:52,040 --> 00:49:53,680 SOME CONCLUDING THOUGHTS. 1196 00:49:53,680 --> 00:49:59,320 THOUGH THE TYPE 2 IMMUNE 1197 00:49:59,320 --> 00:50:02,240 RESPONSE BIASED ECM SCAFFOLD, 1198 00:50:02,240 --> 00:50:04,200 THOUGH THE ECM SCAFFOLD IS TYPE 1199 00:50:04,200 --> 00:50:13,280 2 BY AS DOES NOT SUPPRESS CD8 T 1200 00:50:13,280 --> 00:50:15,400 CELLS, THE CYTO TOXICS IMMUNE 1201 00:50:15,400 --> 00:50:17,560 ADJUVANTS DID NOT NEGATE 1202 00:50:17,560 --> 00:50:23,280 HALLMARK FEATURES OF THE IMMUNE 1203 00:50:23,280 --> 00:50:25,000 RESPONSE AND ECM SCAFFOLD 1204 00:50:25,000 --> 00:50:26,440 SYNERGIZED TO CURE ESTABLISHED 1205 00:50:26,440 --> 00:50:27,440 TUMORS, SO REMEMBER SOLUBLE 1206 00:50:27,440 --> 00:50:31,040 DELIVERY OF THE SAME COMPONENTS 1207 00:50:31,040 --> 00:50:36,040 WAS ONLY A DELAY RATHER THAN A 1208 00:50:36,040 --> 00:50:37,120 CURATIVE RESPONSE. 1209 00:50:37,120 --> 00:50:39,560 AND SO I'LL CONCLUDE SAYING THAT 1210 00:50:39,560 --> 00:50:45,280 CYTOTOXIC T CELL IMMUNITY CAN BE 1211 00:50:45,280 --> 00:50:47,680 BIOORTHOGONAL TO A 1212 00:50:47,680 --> 00:50:48,920 PRO-REGENERATIVE ECM SCAFFOLD 1213 00:50:48,920 --> 00:50:51,080 IMMUNE ENVIRONMENT, WE CAN 1214 00:50:51,080 --> 00:50:54,280 INDUCE ARM OF IMMUNITY WITHOUT 1215 00:50:54,280 --> 00:50:55,240 DAMAGING OR DISRUPTING THE 1216 00:50:55,240 --> 00:50:57,320 BENEFICIAL ELEMENTS OF THAT ECM 1217 00:50:57,320 --> 00:51:03,680 RESPONSE, THAT ARE IMPORTANT FOR 1218 00:51:03,680 --> 00:51:04,360 HEALING, FOR EXAMPLE. 1219 00:51:04,360 --> 00:51:08,920 LET ME THANK MY TEAM FOR MAKING 1220 00:51:08,920 --> 00:51:10,440 THIS ALL POSSIBLE, ESPECIALLY 1221 00:51:10,440 --> 00:51:11,880 SANJAY, WHO DID THE MAJORITY OF 1222 00:51:11,880 --> 00:51:13,200 THIS WORK THAT YOU SAW HERE 1223 00:51:13,200 --> 00:51:21,400 TODAY. 1224 00:51:21,400 --> 00:51:23,600 1225 00:51:23,600 --> 00:51:26,280 WE'RE EXPANDING AND HIRING IF 1226 00:51:26,280 --> 00:51:27,640 YOU KNOW POSTDOCS TO JOIN THE 1227 00:51:27,640 --> 00:51:30,360 LAB AND WORK IN THIS AREA, SEND 1228 00:51:30,360 --> 00:51:31,240 ME A NOTE. 1229 00:51:31,240 --> 00:51:34,280 THANK YOU, AND ARE THERE ANY 1230 00:51:34,280 --> 00:51:34,520 QUESTIONS? 1231 00:51:34,520 --> 00:51:39,240 [APPLAUSE] 1232 00:51:39,240 --> 00:51:39,400 YES? 1233 00:51:39,400 --> 00:51:42,200 >> GREAT TO SEE THE WORK YOU 1234 00:51:42,200 --> 00:51:48,000 PUT TOGETHER. 1235 00:51:48,000 --> 00:51:48,200 1236 00:51:48,200 --> 00:51:49,200 YES, THANK YOU, MATT. 1237 00:51:49,200 --> 00:51:51,480 GREAT TO SEE YOUR WORK PUT 1238 00:51:51,480 --> 00:51:54,080 TOGETHER THIS WAY. 1239 00:51:54,080 --> 00:51:55,000 SO, YOU KNOW, ANTI-COLLAGEN 1240 00:51:55,000 --> 00:51:56,520 ANTIBODIES ARE GENERATED IN 1241 00:51:56,520 --> 00:51:58,240 PEOPLE WITH RHEUMATOID 1242 00:51:58,240 --> 00:51:59,000 ARTHRITIS. 1243 00:51:59,000 --> 00:52:00,920 I'M CURIOUS, USING ECM AS A 1244 00:52:00,920 --> 00:52:02,240 DELIVERY METHOD TO RAMP UP THE 1245 00:52:02,240 --> 00:52:03,960 IMMUNE SYSTEM FOR A VACCINE 1246 00:52:03,960 --> 00:52:04,960 APPROACH, ARE YOU CONCERNED OR 1247 00:52:04,960 --> 00:52:07,600 HAVE YOU CHECKED DID YOU GET 1248 00:52:07,600 --> 00:52:08,720 ANTI-ECM PROTEIN ANTIBODIES THAT 1249 00:52:08,720 --> 00:52:10,240 MIGHT CAUSE SOME OTHER SIDE 1250 00:52:10,240 --> 00:52:12,960 EFFECTS THAT WE WEREN'T LOOKING 1251 00:52:12,960 --> 00:52:13,120 FOR? 1252 00:52:13,120 --> 00:52:15,040 >> YEAH, THAT'S A GOOD 1253 00:52:15,040 --> 00:52:18,000 QUESTION, ONE THAT WE'VE THOUGHT 1254 00:52:18,000 --> 00:52:18,200 ABOUT. 1255 00:52:18,200 --> 00:52:22,920 NOW, HAVEN'T LOOKED FOR -- I 1256 00:52:22,920 --> 00:52:24,160 BELIEVE IT'S ANTI-COLLAGEN TYPE 1257 00:52:24,160 --> 00:52:26,240 2 THAT IS IN RHEUMATOID, BUT, 1258 00:52:26,240 --> 00:52:28,040 NO, WE HAVEN'T SEEN ANY EVIDENCE 1259 00:52:28,040 --> 00:52:33,040 AND WE'VE TAKEN THESE MICE OUT 1260 00:52:33,040 --> 00:52:34,200 FOR ABOUT SIX MONTHS. 1261 00:52:34,200 --> 00:52:36,160 THEY ARE STILL GOING NOW 1262 00:52:36,160 --> 00:52:36,720 ACTUALLY. 1263 00:52:36,720 --> 00:52:40,760 AND THERE'S NO EVIDENCE OF 1264 00:52:40,760 --> 00:52:42,520 DEGENERATION. 1265 00:52:42,520 --> 00:52:44,360 LIKEWISE, WE PERFORMED THIS 1266 00:52:44,360 --> 00:52:46,120 VACCINATION IN MATURE MICE, NOT 1267 00:52:46,120 --> 00:52:50,520 QUITE OLD AGE, WE DIDN'T SEE ANY 1268 00:52:50,520 --> 00:52:52,000 DIFFERENCE OR LONG-TERM 1269 00:52:52,000 --> 00:52:52,680 MORBIDITIES THERE. 1270 00:52:52,680 --> 00:52:54,360 WE DIDN'T LOOK AT JOINTS BUT 1271 00:52:54,360 --> 00:52:55,840 COLLECTED TISSUES ALONG THE WAY 1272 00:52:55,840 --> 00:52:57,800 TO LOOK FOR EVIDENCE OF 1273 00:52:57,800 --> 00:52:58,800 INFLAMMATION IN OTHER TISSUES, 1274 00:52:58,800 --> 00:53:04,040 AND WE JUST DON'T SEE THAT. 1275 00:53:04,040 --> 00:53:06,040 THOUGH FOR THE MOUSE, IT'S A 1276 00:53:06,040 --> 00:53:07,320 CONCERN, BUT KIND OF WHAT MAKES 1277 00:53:07,320 --> 00:53:09,200 ME FEEL BETTER ABOUT THAT TOO IS 1278 00:53:09,200 --> 00:53:11,160 IT'S HARD TO INDUCE THOSE MODELS 1279 00:53:11,160 --> 00:53:15,120 IN THAT YOU NEED A SPECIFIC 1280 00:53:15,120 --> 00:53:16,720 ADJUVANT AND SPECIFIC TYPE OF 1281 00:53:16,720 --> 00:53:17,720 COLLAGEN DELIVERED IN A CERTAIN 1282 00:53:17,720 --> 00:53:23,200 WAY TO MAKE THAT MODEL WORK. 1283 00:53:23,200 --> 00:53:25,600 TO MY KNOWLEDGE. 1284 00:53:25,600 --> 00:53:27,480 AND I THINK THAT THAT MAKES 1285 00:53:27,480 --> 00:53:30,560 SENSE THAT WE'RE ALL VERY 1286 00:53:30,560 --> 00:53:33,600 TOLERANT TO OUR VERY BASE 1287 00:53:33,600 --> 00:53:34,920 STRUCTURAL ECM PROTEINS, THINGS 1288 00:53:34,920 --> 00:53:36,360 IF WE INDUCED AN AUTOIMMUNE 1289 00:53:36,360 --> 00:53:38,640 RESPONSE AGAINST WE'D HAVE A 1290 00:53:38,640 --> 00:53:41,400 DYSTROPHY AND BE IN REALLY BIG 1291 00:53:41,400 --> 00:53:42,280 TROUBLE. 1292 00:53:42,280 --> 00:53:44,720 SO, WE DON'T SEE -- LONG WAY OF 1293 00:53:44,720 --> 00:53:47,640 SAYING WE DON'T SEE IT HAPPENING 1294 00:53:47,640 --> 00:53:48,640 AND SOME THOUGHTS OF WHY THAT 1295 00:53:48,640 --> 00:53:51,160 MIGHT BE. 1296 00:53:51,160 --> 00:53:52,520 1297 00:53:52,520 --> 00:53:57,080 >> FOLLOWING UP ON HIS 1298 00:53:57,080 --> 00:53:58,720 QUESTION, SOMETIMES YOU SEE 1299 00:53:58,720 --> 00:54:01,600 COLLAGEN FORMED AROUND THE 1300 00:54:01,600 --> 00:54:02,880 IMPLANT, THINGS WALLED OFF. 1301 00:54:02,880 --> 00:54:06,720 MY QUESTION RELATES TO YOU HAVE 1302 00:54:06,720 --> 00:54:09,080 MICE THAT SURVIVED AND MICE THAT 1303 00:54:09,080 --> 00:54:10,760 KILL THE TUMORS, T CELLS KILL 1304 00:54:10,760 --> 00:54:11,200 THE TUMORS 1305 00:54:11,200 --> 00:54:14,360 WHAT DO YOU SEE WITH DIFFERENCE 1306 00:54:14,360 --> 00:54:15,480 IN EXTRACELLULAR MATRIX AROUND 1307 00:54:15,480 --> 00:54:18,400 THESE IMPLANTS THAT YOU PUT IN 1308 00:54:18,400 --> 00:54:19,600 AS FAR AS SURVIVAL? 1309 00:54:19,600 --> 00:54:22,440 >>VERY GOOD QUESTION. 1310 00:54:22,440 --> 00:54:25,520 SO, THE BASELINE WITHOUT ANY 1311 00:54:25,520 --> 00:54:27,720 MANIPULATION DIFFERENCE BETWEEN 1312 00:54:27,720 --> 00:54:29,360 THESE MATERIALS AND SOMETHING 1313 00:54:29,360 --> 00:54:30,440 LIKE POLYPROPYLENE MESH IS THAT 1314 00:54:30,440 --> 00:54:33,720 YOU TEND NOT TO GET THAT 1315 00:54:33,720 --> 00:54:36,040 FIBROTIC DEPOSITION WITH THESE 1316 00:54:36,040 --> 00:54:37,440 ECM BIASED MATERIALS, SO THAT'S 1317 00:54:37,440 --> 00:54:39,520 ONE OF THE REASONS THEY ARE USED 1318 00:54:39,520 --> 00:54:44,240 IN THE CLINIC TO AVOID FIBROTIC 1319 00:54:44,240 --> 00:54:47,280 SIDE EFFECTS. 1320 00:54:47,280 --> 00:54:50,360 WE DON'T HAVE LONG-TERM 1321 00:54:50,360 --> 00:54:51,880 EVALUATION OF THE MATERIAL PLUS 1322 00:54:51,880 --> 00:54:53,000 ADJUVANT YET SO THAT'S AN 1323 00:54:53,000 --> 00:54:57,040 INTERESTING QUESTION TO SEE IF 1324 00:54:57,040 --> 00:54:58,120 THAT'S BEING ALTERED, AND IT'S 1325 00:54:58,120 --> 00:55:02,840 ON THE LIST OF THINGS WE'RE 1326 00:55:02,840 --> 00:55:04,360 WORKING ON. 1327 00:55:04,360 --> 00:55:05,760 YEAH, BY DEFAULT, IT SHOULDN'T 1328 00:55:05,760 --> 00:55:08,080 HAPPEN AND WE'RE GOING TO SEE IF 1329 00:55:08,080 --> 00:55:10,320 WE'RE SCREWING THAT UP BY ADDING 1330 00:55:10,320 --> 00:55:18,600 THESE OTHER COMPOUNDS. 1331 00:55:18,600 --> 00:55:22,280 1332 00:55:22,280 --> 00:55:22,600 OKAY. 1333 00:55:22,600 --> 00:55:22,880 THANK YOU. 1334 00:55:22,880 --> 00:55:25,080 [APPLAUSE] 1335 00:55:25,080 --> 00:55:31,120 1336 00:55:31,120 --> 00:55:40,840 1337 00:55:40,840 --> 00:55:51,240 >> TEN-MINUTE BREAK. 1338 01:03:58,640 --> 01:04:01,320 >> WE'RE GETTING BEHIND TIME SO 1339 01:04:01,320 --> 01:04:02,080 LET'S GET GOING AGAIN. 1340 01:04:02,080 --> 01:04:04,480 THANK YOU. 1341 01:04:04,480 --> 01:04:10,520 1342 01:04:10,520 --> 01:04:14,120 1343 01:04:14,120 --> 01:04:15,880 >> ALL RIGHT. 1344 01:04:15,880 --> 01:04:19,280 NEXT SPEAKER, DR. CHEN, IF 1345 01:04:19,280 --> 01:04:21,760 YOU'RE HERE. 1346 01:04:21,760 --> 01:04:24,520 KNOW, NICOLE MORGAN, MY BAD. 1347 01:04:24,520 --> 01:04:26,920 ACTIVE CHIEF OF TRANS-NIH SHARED 1348 01:04:26,920 --> 01:04:28,560 RESOURCE ON BIOMEDICAL 1349 01:04:28,560 --> 01:04:30,440 ENGINEERING AND PHYSICAL 1350 01:04:30,440 --> 01:04:36,000 SCIENCES, RESEARCH FOCUSING ON 1351 01:04:36,000 --> 01:04:37,400 MICROFABRICATION WITH BIOMEDICAL 1352 01:04:37,400 --> 01:04:42,920 RESEARCH, PLEASE WELCOME DR. 1353 01:04:42,920 --> 01:04:43,240 MORGAN. 1354 01:04:43,240 --> 01:04:43,520 [APPLAUSE] 1355 01:04:43,520 --> 01:04:45,760 >> THANKS FOR THE OPPORTUNITY 1356 01:04:45,760 --> 01:04:47,200 TO TALK AND INTRODUCTION. 1357 01:04:47,200 --> 01:04:48,480 I'M GOING TO MOSTLY NOT TALK 1358 01:04:48,480 --> 01:04:50,560 ABOUT MOST OF WHAT WE DO TODAY 1359 01:04:50,560 --> 01:04:53,080 BUT I'M GOING TO TALK ABOUT A 1360 01:04:53,080 --> 01:04:54,200 FEW COLLABORATIONS FOCUSED ON 1361 01:04:54,200 --> 01:04:55,720 DEVELOPING TOOLS FOR CONTROLLING 1362 01:04:55,720 --> 01:04:57,120 AND MEASUREMENT OF OXYGEN IN 1363 01:04:57,120 --> 01:04:59,320 TISSUE CULTURE FOR IN VITRO 1364 01:04:59,320 --> 01:04:59,640 SYSTEMS. 1365 01:04:59,640 --> 01:05:03,160 HOW DO I DO THE THING? 1366 01:05:03,160 --> 01:05:05,360 PERFECT. 1367 01:05:05,360 --> 01:05:05,560 OKAY. 1368 01:05:05,560 --> 01:05:11,800 SO I'M HEAD OF THE SHARED 1369 01:05:11,800 --> 01:05:14,760 RESOURCES, HEAD UP 1370 01:05:14,760 --> 01:05:15,840 MICROFABRICATION AND 1371 01:05:15,840 --> 01:05:17,480 MICROFLUIDICS, WE RUN 1372 01:05:17,480 --> 01:05:19,440 MULTI-USER, IT HELPS TRAINEES 1373 01:05:19,440 --> 01:05:21,320 WITH CURRENT RESEARCH AND FUTURE 1374 01:05:21,320 --> 01:05:24,080 CAREER PROSPECTS TO DEVELOP SOME 1375 01:05:24,080 --> 01:05:26,040 OF THE TECHNIQUES AND TOOLS OF 1376 01:05:26,040 --> 01:05:27,240 BIOMEDICAL ENGINEERING TO TAKE 1377 01:05:27,240 --> 01:05:28,120 WITH THEM. 1378 01:05:28,120 --> 01:05:29,880 IT HELPS US FIGURE OUT A LITTLE 1379 01:05:29,880 --> 01:05:31,920 BIT MORE CLOSELY WHAT PEOPLE 1380 01:05:31,920 --> 01:05:32,720 WANT TO DO REALLY. 1381 01:05:32,720 --> 01:05:37,600 AND IN TERMS OF THE DEVICE 1382 01:05:37,600 --> 01:05:37,880 DESIGN. 1383 01:05:37,880 --> 01:05:39,960 LATER WE'LL TALK MORE ABOUT THE 1384 01:05:39,960 --> 01:05:44,000 FACILITY, I'M GOING TO SKIP TO 1385 01:05:44,000 --> 01:05:45,080 TALKING AND GIVING ORIENTATION 1386 01:05:45,080 --> 01:05:47,000 FOR PROJECTS WE'VE BEEN WORKING 1387 01:05:47,000 --> 01:05:49,560 ON AROUND OXYGEN DELIVERY AND 1388 01:05:49,560 --> 01:05:50,880 CONTROL. 1389 01:05:50,880 --> 01:05:52,400 FOR ORIENTATION, WE'RE BREATHING 1390 01:05:52,400 --> 01:05:55,600 21% OXYGEN RIGHT NOW, THAT'S NOT 1391 01:05:55,600 --> 01:05:58,720 WHAT CELLS ARE EXPERIENCING. 1392 01:05:58,720 --> 01:05:59,880 PHYSIOLOGICAL LEVELS OF OXYGEN 1393 01:05:59,880 --> 01:06:08,360 IN THE CIRCULATION, 3 TO 8% 1394 01:06:08,360 --> 01:06:12,120 PARTIAL PRESSURE, AND 200 1395 01:06:12,120 --> 01:06:17,040 MICRONS, SO IN BETWEEN 1396 01:06:17,040 --> 01:06:22,520 INTERCAPILLARIES LET'S THINK 1397 01:06:22,520 --> 01:06:24,040 ABOUT ADHERENT CELL CULTURE. 1398 01:06:24,040 --> 01:06:28,640 WE HAVE MONOLAYER OF CELLS UNDER 1399 01:06:28,640 --> 01:06:37,520 MEDIA, IN ROOM AIR, 18.6% OXYGEN 1400 01:06:37,520 --> 01:06:45,040 ONCE YOU TAKE CARBON DIOXIDE AND 1401 01:06:45,040 --> 01:06:47,320 WATER VAPOR. 1402 01:06:47,320 --> 01:06:49,400 IT'S SUBSTANTIALLY LOWER THAN 1403 01:06:49,400 --> 01:06:50,040 ATMOSPHERE BECAUSE TRANSPORT IS 1404 01:06:50,040 --> 01:06:52,240 LIMITED IN BY MIXING AND 1405 01:06:52,240 --> 01:06:52,960 DIFFUSION, SUBSTANTIAL DEPLETION 1406 01:06:52,960 --> 01:07:00,040 OF OXYGEN AT THAT CELL SURFACE 1407 01:07:00,040 --> 01:07:01,880 BY CONSUMPTION. 1408 01:07:01,880 --> 01:07:06,360 WE HAVE RECIPES FOR CULTURING. 1409 01:07:06,360 --> 01:07:07,800 THERE ARE OTHER FACTORS AT PLAY 1410 01:07:07,800 --> 01:07:10,320 BUT A LOT HAS TO DO WITH KEEPING 1411 01:07:10,320 --> 01:07:13,800 THEM AT OXYGEN LEVELS THEY ARE 1412 01:07:13,800 --> 01:07:16,040 HAPPY WITH. 1413 01:07:16,040 --> 01:07:19,520 3D THINGS GET COMPLICATED. 1414 01:07:19,520 --> 01:07:21,800 YOU HAVE SPHEROIDS, OTHER 3D 1415 01:07:21,800 --> 01:07:25,960 STRUCTURES IN MATRIGEL OR ECM 1416 01:07:25,960 --> 01:07:34,040 HYDROGEL WITH NO BULK 1417 01:07:34,040 --> 01:07:35,000 CONVECTION, TRANSPORTS DIFFUSION 1418 01:07:35,000 --> 01:07:37,480 ONLY, THE POSITION OF THE 3D 1419 01:07:37,480 --> 01:07:40,160 STRUCTURE IN THE MATRIX, IF YOU 1420 01:07:40,160 --> 01:07:41,600 HAVE A HANGING THAT'S UNIFORM 1421 01:07:41,600 --> 01:07:42,840 BUT YOU HAVE A SITUATION WHERE 1422 01:07:42,840 --> 01:07:46,080 THE OUTSIDE OF THE SPHEROID IS 1423 01:07:46,080 --> 01:07:49,360 PROBABLY AT HIGHER OXYGENATION 1424 01:07:49,360 --> 01:07:49,800 THAN PHYSIOLOGICAL 1425 01:07:49,800 --> 01:07:51,560 CONCENTRATIONS AND INSIDE 1426 01:07:51,560 --> 01:07:54,800 BECOMES HYPOXIC AND NECROTIC. 1427 01:07:54,800 --> 01:07:57,800 ALL SPHEROIDS, YOU DON'T LET 1428 01:07:57,800 --> 01:08:03,680 THEM GET TOO BIG, 500 MICRONS 1429 01:08:03,680 --> 01:08:04,640 CENTERS BECOME NECROTIC. 1430 01:08:04,640 --> 01:08:08,160 YOU HAVE A CULTURE, THE OUTSIDE 1431 01:08:08,160 --> 01:08:12,880 IS LEAST MECHANICALLY 1432 01:08:12,880 --> 01:08:14,280 CONSTRAINED, MOST OXYGENATED, 1433 01:08:14,280 --> 01:08:15,520 CAN MULTIPLY WITHOUT BOUND WHILE 1434 01:08:15,520 --> 01:08:17,800 CELLS IN THE MIDDLE STARVE. 1435 01:08:17,800 --> 01:08:20,440 SO PEOPLE HAVE BEEN WORKING ON 1436 01:08:20,440 --> 01:08:22,960 STRATEGIES FOR OXYGEN DELIVERY 1437 01:08:22,960 --> 01:08:26,400 IN 3D, FASCINATING AND NECESSARY 1438 01:08:26,400 --> 01:08:29,320 WITH TISSUE ON A CHIP, ECM IN 1439 01:08:29,320 --> 01:08:31,400 THE MIDDLE WITH PERFUSION 1440 01:08:31,400 --> 01:08:32,600 CHANNELS, OTHER STRATEGIES WHERE 1441 01:08:32,600 --> 01:08:36,200 PEOPLE WILL DEVELOP SORT OF 1442 01:08:36,200 --> 01:08:39,280 INDUCED VASCULARIZATION INSIDE 1443 01:08:39,280 --> 01:08:41,240 AN ECM, CELL-BEARING ECM. 1444 01:08:41,240 --> 01:08:42,880 AND THESE ARE GREAT BUT HARD TO 1445 01:08:42,880 --> 01:08:43,960 SCALE UP. 1446 01:08:43,960 --> 01:08:46,720 YOU NEED FLOW CONTROL FOR 1447 01:08:46,720 --> 01:08:48,840 MULTIPLE CHANNELS PER DEVICE, 1448 01:08:48,840 --> 01:08:52,440 YOU NEED TO -- IF YOU DON'T 1449 01:08:52,440 --> 01:08:56,040 INDUCE VASCULARIZATION YOU NEED 1450 01:08:56,040 --> 01:08:58,360 PATTERN CHANNELS 200 MICRONS 1451 01:08:58,360 --> 01:08:59,600 APART IN HYDROGEL, STILL HARD. 1452 01:08:59,600 --> 01:09:01,880 YOU HAVE A LARGE PARAMETER 1453 01:09:01,880 --> 01:09:04,280 STAYS, CHOOSE USUAL ECM, CELL 1454 01:09:04,280 --> 01:09:06,920 TYPE, FLOW CONDITIONS, SO IT'S 1455 01:09:06,920 --> 01:09:09,000 HARD TO BE CONFIDENT THAT THE 1456 01:09:09,000 --> 01:09:10,320 PARTICULAR CHOICES YOU'VE MADE 1457 01:09:10,320 --> 01:09:11,640 ALONG THOSE DIMENSIONS ARE GOING 1458 01:09:11,640 --> 01:09:15,000 TO BE RELEVANT FOR THE BIOLOGY 1459 01:09:15,000 --> 01:09:16,440 YOU WANT. 1460 01:09:16,440 --> 01:09:18,480 SO, WHEN WE LOOKED AT THIS 1461 01:09:18,480 --> 01:09:21,200 PROBLEM, WHAT IS THE SIMPLEST 1462 01:09:21,200 --> 01:09:23,440 WAY THAT'S SCALABLE THAT WE CAN 1463 01:09:23,440 --> 01:09:24,720 GET CONTROLLED OXYGEN DELIVERY 1464 01:09:24,720 --> 01:09:26,160 TO 3D CULTURE VOLUMES. 1465 01:09:26,160 --> 01:09:29,880 SO THE SYSTEM WE HAVE IS THIS 1466 01:09:29,880 --> 01:09:32,080 OXYGEN PERMEABLE MEMBRANE WITH 1467 01:09:32,080 --> 01:09:35,800 PILLARS SPACED BY INTERCAPILLARY 1468 01:09:35,800 --> 01:09:37,320 DISTANCE, 200 MICRONS APART, 1469 01:09:37,320 --> 01:09:40,520 SEED CELLS AROUND THESE PILLARS 1470 01:09:40,520 --> 01:09:43,320 IN MATRIGEL, PUT THE WHOLE 1471 01:09:43,320 --> 01:09:45,840 VESSEL INSIDE THIS BIOREACTOR 1472 01:09:45,840 --> 01:09:50,000 THAT HAS -- HOW DOES THE POINTER 1473 01:09:50,000 --> 01:09:50,200 WORK? 1474 01:09:50,200 --> 01:09:50,680 INTERESTING. 1475 01:09:50,680 --> 01:09:58,200 WE PUT THE WHOLE SYSTEM IN A 1476 01:09:58,200 --> 01:10:00,080 BIOREACTOR WITH TWO 1477 01:10:00,080 --> 01:10:02,720 CONTROLLED -- TWO CHAMBERS WITH 1478 01:10:02,720 --> 01:10:04,040 CONTROLLED ATMOSPHERE, SOURCE 1479 01:10:04,040 --> 01:10:07,080 GAS ON THE BOTTOM, 3% OXYGEN IF 1480 01:10:07,080 --> 01:10:08,280 WE'RE LOOKING FOR PHYSIOLOGICAL, 1481 01:10:08,280 --> 01:10:10,040 LOWER IF WE WANT HYPOXIA. 1482 01:10:10,040 --> 01:10:11,680 THERE'S MEDIA ON TOP. 1483 01:10:11,680 --> 01:10:16,040 ON TOP OF THAT SPACE IT'S 1484 01:10:16,040 --> 01:10:19,880 ANOXIC, 5% CO2 FOR PH BALANCE 1485 01:10:19,880 --> 01:10:20,320 AND NITROGEN. 1486 01:10:20,320 --> 01:10:23,800 THIS IS WHAT THE BIOREACTOR 1487 01:10:23,800 --> 01:10:25,560 LOOKS LIKE FIRST GENERATION FOR 1488 01:10:25,560 --> 01:10:30,240 MULTI-WELL PLATES, AGAIN TWO 1489 01:10:30,240 --> 01:10:32,560 CHAMBERS, SIX SIX-WELL PLATES, 1490 01:10:32,560 --> 01:10:34,200 GET TO HIGHER DENSITY OF PLATES 1491 01:10:34,200 --> 01:10:38,040 OR HIGHER DENSITY OF WELLS. 1492 01:10:38,040 --> 01:10:44,400 THERE'S LIKE TWO MACHINE TO 1493 01:10:44,400 --> 01:10:45,240 PLASTIC PIECES SANDWICH BE 1494 01:10:45,240 --> 01:10:48,680 MEMBRANE IN BETWEEN. 1495 01:10:48,680 --> 01:10:52,480 THERE'S A CONTROL CHAMBER BELT 1496 01:10:52,480 --> 01:10:56,080 IN -- BUILT IN, YOU CAN USE IT 1497 01:10:56,080 --> 01:10:58,080 AT THE SAME TIME YOU RUN THE 1498 01:10:58,080 --> 01:10:58,440 EXPERIMENT. 1499 01:10:58,440 --> 01:11:00,480 WHAT DO WE GET HERE? 1500 01:11:00,480 --> 01:11:02,320 WHAT WE'LL SAY IS CONVENTIONAL 1501 01:11:02,320 --> 01:11:03,960 IS LEFT TWO COLUMNS, CELLS GROWN 1502 01:11:03,960 --> 01:11:06,520 WITH 21% OXYGEN FROM THE TOP AND 1503 01:11:06,520 --> 01:11:08,640 3% FROM THE TOP, SORT OF THE 1504 01:11:08,640 --> 01:11:11,200 HYPOXIA CHAMBER, IF YOU WILL. 1505 01:11:11,200 --> 01:11:14,080 AND YOU SEE WHAT YOU EXPECT, 1506 01:11:14,080 --> 01:11:16,040 GROWING MORE HAPPILY IN 21% THAN 1507 01:11:16,040 --> 01:11:17,000 3%. 1508 01:11:17,000 --> 01:11:20,040 WHEN YOU PUT CELLS IN THE 1509 01:11:20,040 --> 01:11:22,480 GRADIENT CONFIGURATION IN 1510 01:11:22,480 --> 01:11:25,440 BETWEEN CHAMBERS, 3% AND 0% 1511 01:11:25,440 --> 01:11:33,880 OXYGEN, THEY GROW AS WELL AS 2% 1512 01:11:33,880 --> 01:11:38,240 TO A WEEK, SEE MORE HYPOXIA IN 1513 01:11:38,240 --> 01:11:39,440 21% CELLS, WHAT'S HAPPENING IS 1514 01:11:39,440 --> 01:11:41,440 THE CELLS NEAR THE OXYGEN HAVE 1515 01:11:41,440 --> 01:11:47,760 PROLIFERATED TO THE POINT THEY 1516 01:11:47,760 --> 01:11:48,200 ARE STARVING. 1517 01:11:48,200 --> 01:11:50,200 AND SO YOU ARE -- I'M GOING TO 1518 01:11:50,200 --> 01:11:52,240 MOVE ON BUT YOU'RE LOOK TO LOOK 1519 01:11:52,240 --> 01:11:53,800 AT MORE GENERAL EXPRESSION 1520 01:11:53,800 --> 01:11:56,040 ANALYSIS IN THE PUBLISHED 1521 01:11:56,040 --> 01:11:59,040 RESULTS BUT, AGAIN, WHAT YOU 1522 01:11:59,040 --> 01:12:01,240 WOULD EXPECT FROM FROM PRINCIPAL 1523 01:12:01,240 --> 01:12:03,880 COMPONENT ANALYSIS WHERE THE 1524 01:12:03,880 --> 01:12:06,280 GRADIENT CELLS, 21% CLUSTER MORE 1525 01:12:06,280 --> 01:12:08,240 TIGHTLY, 3% CELLS ARE WHAT WE 1526 01:12:08,240 --> 01:12:11,120 WOULD THINK OF MORE AS A HYPOXIC 1527 01:12:11,120 --> 01:12:14,120 SAMPLE, AND THEN AS A PLUG TO -- 1528 01:12:14,120 --> 01:12:16,120 PLEASE DO CULTURE EXPERIMENTS IN 1529 01:12:16,120 --> 01:12:18,880 3D, 2D IS IN LEFT FIELD. 1530 01:12:18,880 --> 01:12:21,480 SO, WHAT ARE WE TRYING TO DO 1531 01:12:21,480 --> 01:12:21,800 NOW? 1532 01:12:21,800 --> 01:12:23,800 WE'RE LOOKING AT NOW BIOLOGICAL 1533 01:12:23,800 --> 01:12:26,280 QUESTIONS LIKE DRUG RESISTANCE, 1534 01:12:26,280 --> 01:12:27,280 FUNCTION OF OXYGENATION, MONITOR 1535 01:12:27,280 --> 01:12:30,880 GROWTH SO WE HAVE DEVICES THAT 1536 01:12:30,880 --> 01:12:32,080 DELIVER OXYGEN IN SIMILAR 1537 01:12:32,080 --> 01:12:33,520 CONFIGURATIONS COMPATIBLE WITH 1538 01:12:33,520 --> 01:12:34,640 LONG-TERM IMAGING MICROSCOPES 1539 01:12:34,640 --> 01:12:36,080 AND LOOKING FOR SPATIAL 1540 01:12:36,080 --> 01:12:36,360 RECEIPTOUTS. 1541 01:12:36,360 --> 01:12:41,000 WHAT I WANT TO TALK ABOUT NOW IS 1542 01:12:41,000 --> 01:12:44,920 WHETHER WE CAN ALSO IN REAL TIME 1543 01:12:44,920 --> 01:12:45,360 CHARACTERIZE OXYGEN 1544 01:12:45,360 --> 01:12:45,960 CONCENTRATION EXPERIENCED BY 1545 01:12:45,960 --> 01:12:52,160 CELLS AS FUNCTION OF DELIVERING 1546 01:12:52,160 --> 01:12:52,680 CONSUMPTION. 1547 01:12:52,680 --> 01:12:55,840 WE STARTED WITH A COLLABORATION 1548 01:12:55,840 --> 01:13:01,320 WITH NHLBI, WITH A PROBE FOR 1549 01:13:01,320 --> 01:13:03,320 MEASURING INTRACELLULAR OXYGEN, 1550 01:13:03,320 --> 01:13:07,680 A FUSION, THE OVERLAP OF 1551 01:13:07,680 --> 01:13:09,680 mCHERRY AND MYOGLOBIN DEPENDS 1552 01:13:09,680 --> 01:13:14,680 ON OXYGENATION STATE OF 1553 01:13:14,680 --> 01:13:14,960 MYOGLOBIN. 1554 01:13:14,960 --> 01:13:17,320 SO, HOW WE MEASURE THIS IS WITH 1555 01:13:17,320 --> 01:13:17,560 LIFETIME. 1556 01:13:17,560 --> 01:13:19,600 WE COULD MEASURE A CHANGE IN 1557 01:13:19,600 --> 01:13:20,480 INTENSITY, THAT'S GOING TO 1558 01:13:20,480 --> 01:13:22,120 DEPEND ON EXPRESSION OF THE 1559 01:13:22,120 --> 01:13:23,680 PROBE AND CELLS, THIS IS A MESS. 1560 01:13:23,680 --> 01:13:25,840 WE'RE GOING TO MEASURE LIFETIME, 1561 01:13:25,840 --> 01:13:26,800 INDEPENDENT OF CONCENTRATION. 1562 01:13:26,800 --> 01:13:29,000 HOW THIS WORKS, WE EXCITE WITH 1563 01:13:29,000 --> 01:13:30,840 LASER PULSE, MEASURE THE PHOTONS 1564 01:13:30,840 --> 01:13:33,040 EMITTED AS A FUNCTION OF TIME, 1565 01:13:33,040 --> 01:13:36,560 DO THIS MANY TIMES, BUILD UP A 1566 01:13:36,560 --> 01:13:37,520 DISTRIBUTION OF PHOTONS EMITTED, 1567 01:13:37,520 --> 01:13:47,840 AND WE GO AHEAD AND WE FIT THAT 1568 01:13:47,840 --> 01:13:49,120 DISTRIBUTION, AS EXPONENTIAL 1569 01:13:49,120 --> 01:13:49,720 DECAY. 1570 01:13:49,720 --> 01:13:54,960 AND WE'RE IMAGING SO REPEAT FOR 1571 01:13:54,960 --> 01:14:02,920 EACH PIXEL. 1572 01:14:02,920 --> 01:14:04,480 WE'RE MEASURING INTRINSIC 1573 01:14:04,480 --> 01:14:05,800 FLUORESCENCE, COMBINED WITH 1574 01:14:05,800 --> 01:14:06,560 OXYGEN CONCENTRATION 1575 01:14:06,560 --> 01:14:07,200 MEASUREMENTS. 1576 01:14:07,200 --> 01:14:09,960 AND QUICKLY WE'RE USING TWO 1577 01:14:09,960 --> 01:14:12,040 PHOTON EXCITATION TO MAKE SURE 1578 01:14:12,040 --> 01:14:14,000 THAT WE'RE GETTING MINIMAL 1579 01:14:14,000 --> 01:14:15,400 BACKGROUND FLUORESCENCE AND LESS 1580 01:14:15,400 --> 01:14:16,280 PHOTO DAMAGE. 1581 01:14:16,280 --> 01:14:20,680 THIS IS WHAT WE SEE. 1582 01:14:20,680 --> 01:14:23,960 THIS IS CELLS, ADHERENT CELLS 1583 01:14:23,960 --> 01:14:28,120 MEASURED WITH ATMOSPHERIC OXYGEN 1584 01:14:28,120 --> 01:14:30,400 AND THEN LOWEST OXYGEN THE 1585 01:14:30,400 --> 01:14:34,160 CHAMBER WILL GET TO, 5%. 1586 01:14:34,160 --> 01:14:37,000 WITH THE STANDARD MYOmCHERRY 1587 01:14:37,000 --> 01:14:40,600 PROBE, RIGHT PANELS TARGETED TO 1588 01:14:40,600 --> 01:14:41,800 MITOCHONDRIA. 1589 01:14:41,800 --> 01:14:44,520 WHAT WE DO TO CONVERT THE 1590 01:14:44,520 --> 01:14:45,720 LIFETIME, EVENTUALLY I'LL FIGURE 1591 01:14:45,720 --> 01:14:47,160 THIS OUT, BUT MAYBE NOT TODAY. 1592 01:14:47,160 --> 01:14:50,760 WHAT WE SEE WITH THE LIFETIME -- 1593 01:14:50,760 --> 01:14:50,920 OH. 1594 01:14:50,920 --> 01:14:53,400 WHAT WE SEE WITH THE LIFETIME IS 1595 01:14:53,400 --> 01:14:58,880 HOW WE GET -- LET ME TRY AGAIN. 1596 01:14:58,880 --> 01:15:04,360 HOW WE GO FROM THE LIFETIME TO 1597 01:15:04,360 --> 01:15:12,000 OXYGEN CONCENTRATION IS WE HALT 1598 01:15:12,000 --> 01:15:17,360 IN THE CELLS, THE TOP PANEL, 1599 01:15:17,360 --> 01:15:19,000 ASSUMPTION THAT INSIDE IS THE 1600 01:15:19,000 --> 01:15:21,200 SAME AS OUTSIDE MEASURED WITH 1601 01:15:21,200 --> 01:15:21,880 FIBEROPTIC PROBE. 1602 01:15:21,880 --> 01:15:25,680 USING THAT DATA WE'RE ABLE TO 1603 01:15:25,680 --> 01:15:27,960 GET THE COMPARTMENTAL OXYGEN AS 1604 01:15:27,960 --> 01:15:32,680 A FUNCTION OF THE EXTRACELLULAR 1605 01:15:32,680 --> 01:15:35,640 OXYGEN IN THE RESPIRING CELLS. 1606 01:15:35,640 --> 01:15:36,480 WE'RE THE MICROFABRICATION 1607 01:15:36,480 --> 01:15:38,240 GROUP. 1608 01:15:38,240 --> 01:15:41,960 WHAT IF WE WERE ABLE TO GET 1609 01:15:41,960 --> 01:15:42,960 MEASUREMENTS OF THE OXYGEN 1610 01:15:42,960 --> 01:15:44,680 CONCENTRATION IN THESE CELLS AS 1611 01:15:44,680 --> 01:15:52,040 FUNCTION OF SUPPLIED OXYGEN 1612 01:15:52,040 --> 01:15:54,000 CONCENTRATION FOR A RANGE, MADE 1613 01:15:54,000 --> 01:15:56,440 A SUBSTRATE WITH A SINGLE GAS 1614 01:15:56,440 --> 01:15:58,160 CHANNEL WE FEED 8% OXYGEN 1615 01:15:58,160 --> 01:16:01,560 THROUGH AND THIN LAYER OF PDMS, 1616 01:16:01,560 --> 01:16:04,320 SO THE OTHER BLUE BOXES ARE JUST 1617 01:16:04,320 --> 01:16:07,680 A RULE TO USE THE MICROSCOPE, 1618 01:16:07,680 --> 01:16:13,160 FIGURE OUT WHERE WE ARE, 1619 01:16:13,160 --> 01:16:13,960 MICROSCOPIC IMAGES. 1620 01:16:13,960 --> 01:16:16,560 SO, WHAT WE GET IF WE ARE THEN 1621 01:16:16,560 --> 01:16:17,600 LOOKING AT THE OXYGEN 1622 01:16:17,600 --> 01:16:19,040 CONCENTRATION AS A FUNCTION OF 1623 01:16:19,040 --> 01:16:20,600 DISTANCE FROM THE SOURCE CHANNEL 1624 01:16:20,600 --> 01:16:25,720 WE HAVE MEASURED ON THE LEFT, 1625 01:16:25,720 --> 01:16:27,480 MODELED ON THE RIGHT. 1626 01:16:27,480 --> 01:16:28,320 DON'T HAVE CONVECTION. 1627 01:16:28,320 --> 01:16:34,040 THIS IS PRETTY GOOD AGREEMENT. 1628 01:16:34,040 --> 01:16:35,560 MEASUREMENTS WITH FIBEROPTIC 1629 01:16:35,560 --> 01:16:37,880 PROBE AND GET REASONABLE 1630 01:16:37,880 --> 01:16:39,080 AGREEMENT, A GOOD GRADIENT AND 1631 01:16:39,080 --> 01:16:45,640 CAN PUT CELLS IN THIS AND 1632 01:16:45,640 --> 01:16:49,800 MEASURE OUR MYO-mCHERRY AND 1633 01:16:49,800 --> 01:16:52,000 AUTOFLUORESCENCE AND SEE HIGHER 1634 01:16:52,000 --> 01:16:53,640 OXYGENATION CLOSE TO SOURCE 1635 01:16:53,640 --> 01:16:55,200 CHANNEL, LESS OXYGENATION AWAY 1636 01:16:55,200 --> 01:16:56,840 FROM SOURCE CHANNEL, MORE 1637 01:16:56,840 --> 01:16:59,320 GLYCOLYSIS FARTHER FROM THE 1638 01:16:59,320 --> 01:17:01,920 SOURCE CHANNEL. 1639 01:17:01,920 --> 01:17:04,400 THIS IS CONSISTENT THROUGH 1640 01:17:04,400 --> 01:17:05,480 MULTIPLE MEASUREMENTS. 1641 01:17:05,480 --> 01:17:10,240 AND THEN WHAT I'LL SAY WE CAN DO 1642 01:17:10,240 --> 01:17:12,920 WITH THIS TECHNIQUE, THIS IS NOT 1643 01:17:12,920 --> 01:17:16,160 A GREAT EXAMPLE, WE ALSO GET 1644 01:17:16,160 --> 01:17:17,200 NON-EQUILIBRIUM DATA. 1645 01:17:17,200 --> 01:17:18,720 WHAT I SHOWED UP TILL NOW WHAT 1646 01:17:18,720 --> 01:17:20,360 HAPPENS IF YOU WAIT TWO HOURS 1647 01:17:20,360 --> 01:17:21,000 AND START MEASURING. 1648 01:17:21,000 --> 01:17:22,360 THIS IS WHAT HAPPENS IF YOU 1649 01:17:22,360 --> 01:17:24,520 START MEASURING RIGHT AWAY. 1650 01:17:24,520 --> 01:17:26,920 SO YOU CAN ACTUALLY -- THIS IS 1651 01:17:26,920 --> 01:17:29,040 SOMETHING WHERE WE JUST CHANGE 1652 01:17:29,040 --> 01:17:32,920 THE CONCENTRATION IN ATMOSPHER, 1653 01:17:32,920 --> 01:17:35,120 ALSO COULD DO IN RESPONSE TO 1654 01:17:35,120 --> 01:17:37,520 ADDING A DRUG OR CHANGING MEDIA 1655 01:17:37,520 --> 01:17:39,280 AND MEASURE RESPONSES IN REAL 1656 01:17:39,280 --> 01:17:39,520 TIME. 1657 01:17:39,520 --> 01:17:42,720 THE OTHER THING WE WOULD LIKE TO 1658 01:17:42,720 --> 01:17:45,200 DO BUT NEED TO UNDERSTAND 1659 01:17:45,200 --> 01:17:46,120 SPATIAL AVERAGING SOFTWARE 1660 01:17:46,120 --> 01:17:47,520 BETTER, SO IF PEOPLE HAVE 1661 01:17:47,520 --> 01:17:48,800 SUGGESTIONS THAT WOULD BE GREAT, 1662 01:17:48,800 --> 01:17:50,080 WE WITH LOOK AT HETEROGENEITY. 1663 01:17:50,080 --> 01:17:53,200 IF YOU LOOKED AT IMAGES BEFORE, 1664 01:17:53,200 --> 01:17:55,400 WHAT WE SAW WAS THAT THEY 1665 01:17:55,400 --> 01:17:57,800 WEREN'T ALL THE SAME COLOR SO WE 1666 01:17:57,800 --> 01:18:02,720 HAVE OTHER TECHNIQUES THAT WE 1667 01:18:02,720 --> 01:18:04,880 MEASURE ENSEMBLE AVERAGES OF 1668 01:18:04,880 --> 01:18:05,520 CONSUMPTION RATE OXYGEN 1669 01:18:05,520 --> 01:18:07,000 CONCENTRATION BUT DON'TS HAVE 1670 01:18:07,000 --> 01:18:08,760 THE SORT OF GRANULAR DATE WE 1671 01:18:08,760 --> 01:18:10,280 HAVE GET FROM THAT. 1672 01:18:10,280 --> 01:18:11,880 WE'RE INTERESTED IN LOOKING AT 1673 01:18:11,880 --> 01:18:13,320 HETEROGENEITY WITHIN A LAYER AND 1674 01:18:13,320 --> 01:18:14,600 REALLY WHAT WE WANT TO DO OF 1675 01:18:14,600 --> 01:18:16,400 COURSE IS TO SEE WHAT HAPPENS 1676 01:18:16,400 --> 01:18:18,360 WHEN WE GO INTO 3D. 1677 01:18:18,360 --> 01:18:20,520 THESE ARE PRELIMINARY 1678 01:18:20,520 --> 01:18:21,200 MEASUREMENTS ON SPHEROIDS, 1679 01:18:21,200 --> 01:18:23,000 CHANGED COLOR SCALE ON YOU, WE 1680 01:18:23,000 --> 01:18:26,480 CAN GET A FEW HUNDRED MICRONS 1681 01:18:26,480 --> 01:18:27,680 INTO THE SPHEROID WITH 1682 01:18:27,680 --> 01:18:30,040 SINGLE-CELL RESOLUTION AND 1683 01:18:30,040 --> 01:18:31,280 MEASURE OXYGEN CONCENTRATION IN 1684 01:18:31,280 --> 01:18:32,480 THAT SYSTEM AS WELL. 1685 01:18:32,480 --> 01:18:34,120 AND WHAT WE WANT TO DO NOW IS 1686 01:18:34,120 --> 01:18:36,440 SORT OF PUT THAT -- COMBINE THAT 1687 01:18:36,440 --> 01:18:39,840 WITH THE OTHER SYSTEM THAT WE 1688 01:18:39,840 --> 01:18:40,360 HAVE. 1689 01:18:40,360 --> 01:18:40,880 OKAY. 1690 01:18:40,880 --> 01:18:48,440 SO, ONE QUICK DETOUR ON THIS, 1691 01:18:48,440 --> 01:18:50,400 MYO-mCHERRY SYSTEM, WE HAD TWO 1692 01:18:50,400 --> 01:18:51,840 PROBES, ONE TARGETED TO 1693 01:18:51,840 --> 01:18:53,800 CYTOPLASM, ONE TO MITOCHONDRIA. 1694 01:18:53,800 --> 01:18:56,320 AND WE SEE A MEASURABLE 1695 01:18:56,320 --> 01:18:57,640 DIFFERENCE IN OXYGEN 1696 01:18:57,640 --> 01:19:01,160 CONCENTRATION BETWEEN THE TWO 1697 01:19:01,160 --> 01:19:01,640 COMPARTMENTS. 1698 01:19:01,640 --> 01:19:02,800 AND OUR COLLABORATORS LAB KEPT 1699 01:19:02,800 --> 01:19:05,080 GETTING DINGED WITH THIS 1700 01:19:05,080 --> 01:19:07,800 ASSERTION, WELL, YOU CAN'T HAVE 1701 01:19:07,800 --> 01:19:08,840 THESE SUBCELLULAR DIFFERENCES IN 1702 01:19:08,840 --> 01:19:10,560 OXYGEN CONCENTRATION, THIS 1703 01:19:10,560 --> 01:19:11,040 DOESN'T HAPPEN. 1704 01:19:11,040 --> 01:19:15,480 AND SO WE KEPT LOOKING AND 1705 01:19:15,480 --> 01:19:16,480 SEEING IT. 1706 01:19:16,480 --> 01:19:18,680 AND SO WE DECIDED TO TAKE A LOOK 1707 01:19:18,680 --> 01:19:22,080 AT IT WITH FINITE ELEMENT 1708 01:19:22,080 --> 01:19:22,600 MODELING. 1709 01:19:22,600 --> 01:19:24,480 I HAD A POSTBAC DURING COVID AND 1710 01:19:24,480 --> 01:19:28,280 BEYOND THAT WHO WENT AHEAD AND 1711 01:19:28,280 --> 01:19:29,720 USED MICROSCOPIC IMAGE OF 1712 01:19:29,720 --> 01:19:34,320 MITOCHONDRIAL NETWORK IN A MOUSE 1713 01:19:34,320 --> 01:19:35,960 MYOBLAST TO CONSTRUCT GEOMETRIC 1714 01:19:35,960 --> 01:19:37,920 MODEL SIMILAR IN SCALE AND 1715 01:19:37,920 --> 01:19:41,520 COMPLEXITY AND CONNECTIVITY AND 1716 01:19:41,520 --> 01:19:43,480 PUT IT IN A QUARTER CELL AND 1717 01:19:43,480 --> 01:19:45,720 WENT AHEAD AND USED THAT WITH 1718 01:19:45,720 --> 01:19:47,840 LITERATURE VALUES TO SEE IF HE 1719 01:19:47,840 --> 01:19:58,320 COULD LOOK AT ON THE FORMATION 1720 01:19:59,440 --> 01:20:00,680 OF SUBCELLULAR OXYGEN GRADIENTS, 1721 01:20:00,680 --> 01:20:06,240 CELLS GROWING ON IMPERMEABLE 1722 01:20:06,240 --> 01:20:07,480 SURFACE, GLASS. 1723 01:20:07,480 --> 01:20:09,400 THE COLOR SCALE IS NORMALIZED 1724 01:20:09,400 --> 01:20:10,960 BUT IF YOU LOOK AT THE CELLS 1725 01:20:10,960 --> 01:20:14,600 BEING GROWN IN THE LOW OXYGEN 1726 01:20:14,600 --> 01:20:15,280 CONDITION, SORRY, GO BACK. 1727 01:20:15,280 --> 01:20:18,640 IF YOU LOOK AT THE CELLS IN THE 1728 01:20:18,640 --> 01:20:20,440 LOW OXYGEN CONDITION, THERE'S A 1729 01:20:20,440 --> 01:20:23,440 MUCH -- THERE'S DEPLETION OVER A 1730 01:20:23,440 --> 01:20:33,920 GREATER SPATIAL EXTENT THAN 1731 01:20:36,960 --> 01:20:38,120 ATMOSPHERIC OXYGEN. 1732 01:20:38,120 --> 01:20:41,240 CELL DENSITY HAS A MAJOR EFFECT. 1733 01:20:41,240 --> 01:20:44,040 THIS IS THE DIAGONAL OF A 1734 01:20:44,040 --> 01:20:47,400 COMPUTATIONAL CELL, CELLS ON A 1735 01:20:47,400 --> 01:20:49,160 SQUARE ARRAY, AND IF YOU HAVE 1736 01:20:49,160 --> 01:20:53,640 CELLS THAT ARE CLOSELY SPACED 1737 01:20:53,640 --> 01:20:57,040 THEY DEVELOP SUBSTANTIAL 1738 01:20:57,040 --> 01:20:58,480 GRADIENT IN OXYGEN 1739 01:20:58,480 --> 01:20:59,760 CONSTELLATION, FARTHER APART 1740 01:20:59,760 --> 01:21:03,920 THAN CELL DIAMETER YOU'RE VERY 1741 01:21:03,920 --> 01:21:05,120 CLOSE TO EQUILIBRIUM ATMOSPHERE 1742 01:21:05,120 --> 01:21:08,200 VALUES, JUST A LITTLE DISTANCE 1743 01:21:08,200 --> 01:21:12,680 AWAY FROM THE CELLS. 1744 01:21:12,680 --> 01:21:14,640 AND THEN FINALLY THE -- RELEVANT 1745 01:21:14,640 --> 01:21:18,680 TO BOTH CAN YOU HAVE SUBCELLULAR 1746 01:21:18,680 --> 01:21:19,920 GRADIENTS, CAN YOU HAVE 1747 01:21:19,920 --> 01:21:22,000 SUBSTANTIAL GRADIENTS IN YOUR 1748 01:21:22,000 --> 01:21:24,400 CELLS AND EARLIER BIOREACTOR 1749 01:21:24,400 --> 01:21:24,840 WORK? 1750 01:21:24,840 --> 01:21:26,040 WHETHER THE SURFACE YOU'RE 1751 01:21:26,040 --> 01:21:27,240 GROWING CELLS ON IS PERMEABLE TO 1752 01:21:27,240 --> 01:21:29,440 OXYGEN OR NOT MAKES A HUGE 1753 01:21:29,440 --> 01:21:32,040 DIFFERENCE, SO WE HAVE ON THE 1754 01:21:32,040 --> 01:21:33,920 LEFT CELLS GROWING ON GLASS, ON 1755 01:21:33,920 --> 01:21:37,640 THE RIGHT ON PDMS WITH OXYGEN 1756 01:21:37,640 --> 01:21:40,600 FROM BELOW AS WELL, AND YOU'LL 1757 01:21:40,600 --> 01:21:45,320 NOTICE ONE IS THE GRADE WENTZ 1758 01:21:45,320 --> 01:21:47,960 SEEN IN THE PDMS SITUATION 1759 01:21:47,960 --> 01:21:49,800 DEVICE ARE MUCH SMALLER, YOU CAN 1760 01:21:49,800 --> 01:21:52,880 SEE THE NUMBERS IN THE CHART 1761 01:21:52,880 --> 01:21:53,080 BELOW. 1762 01:21:53,080 --> 01:21:54,800 YOU CAN SEE THAT IN THE NUMBERS 1763 01:21:54,800 --> 01:21:59,080 IN THE CHART BELOW AS WELL, 1764 01:21:59,080 --> 01:22:00,840 DIFFERENCES BETWEEN TWO 1765 01:22:00,840 --> 01:22:02,400 COMPARTMENTS, BUT ALSO THE 1766 01:22:02,400 --> 01:22:04,800 OXYGEN CONCENTRATION THE CELL IS 1767 01:22:04,800 --> 01:22:06,320 EXPERIENCING IN PDMS IS THE 1768 01:22:06,320 --> 01:22:12,920 SOURCE CON CONCENTRATION, 1769 01:22:12,920 --> 01:22:14,200 CONSISTENT WITH RESULTS WHERE WE 1770 01:22:14,200 --> 01:22:15,960 HAVE A SOURCE OXYGEN 1771 01:22:15,960 --> 01:22:18,360 CONCENTRATION OF 3%, CELLS 1772 01:22:18,360 --> 01:22:18,760 GROWING HAPPILY. 1773 01:22:18,760 --> 01:22:20,840 SO I THINK THAT IS MY TIME. 1774 01:22:20,840 --> 01:22:23,080 AND I JUST WANT TO THANK 1775 01:22:23,080 --> 01:22:24,800 EVERYONE WHO HELPED WORK ON THIS 1776 01:22:24,800 --> 01:22:29,440 SO IT'S A COLLABORATION BETWEEN 1777 01:22:29,440 --> 01:22:31,040 OUR GROUP AND MICROFABRICATION 1778 01:22:31,040 --> 01:22:37,640 UNIT, AND MICHAEL GOTTESMAN'S 1779 01:22:37,640 --> 01:22:47,920 GROUP, AND OTHERS. 1780 01:22:53,800 --> 01:22:54,160 THANKS. 1781 01:22:54,160 --> 01:22:56,360 [APPLAUSE] 1782 01:22:56,360 --> 01:23:02,360 1783 01:23:02,360 --> 01:23:04,000 1784 01:23:04,000 --> 01:23:05,320 >> THANK YOU. 1785 01:23:05,320 --> 01:23:06,960 THIS IS REALLY EXCITING. 1786 01:23:06,960 --> 01:23:09,800 I HAVE TWO QUESTIONS. 1787 01:23:09,800 --> 01:23:17,480 ONE, SINCE YOU TAKE THE 1788 01:23:17,480 --> 01:23:20,960 mCHERRY LABELED CELLS, HAVE YOU 1789 01:23:20,960 --> 01:23:22,920 BEEN ABLE TO DO CORRELATIONS IN 1790 01:23:22,920 --> 01:23:23,560 THE SIGNAL? 1791 01:23:23,560 --> 01:23:27,960 SO IF YOU SEE CHANGES IN 1792 01:23:27,960 --> 01:23:32,640 mCHERRY DO YOU SEE -- 1793 01:23:32,640 --> 01:23:35,200 >>IN THE ONE SLIDE I HAD ON 1794 01:23:35,200 --> 01:23:37,240 HETEROGENEITY WE CAN TALK LATER, 1795 01:23:37,240 --> 01:23:39,320 BUT THE ONE SLIDE ON 1796 01:23:39,320 --> 01:23:41,080 HETEROGENEITY THAT WAS 1797 01:23:41,080 --> 01:23:45,160 CORRELATION, SORT OF LOOKING AT 1798 01:23:45,160 --> 01:23:49,200 THE TWO AXES, MYO-mCHERRY AND 1799 01:23:49,200 --> 01:23:51,200 AUTOFLUORESCENCE FOR DIFFERENT 1800 01:23:51,200 --> 01:23:51,920 REGIONS. 1801 01:23:51,920 --> 01:23:56,760 IT'S -- I THINK -- IT'S OKAY. 1802 01:23:56,760 --> 01:23:58,320 YEAH, WHAT WE -- I THINK I WOULD 1803 01:23:58,320 --> 01:24:00,720 LIKE TO BE A LITTLE BIT MORE 1804 01:24:00,720 --> 01:24:01,680 CONFIDENT IN THE SPATIAL 1805 01:24:01,680 --> 01:24:03,200 AVERAGING OF THE SOFTWARE, SO I 1806 01:24:03,200 --> 01:24:05,520 NEED TO DO A BIT OF DEEP DIVE 1807 01:24:05,520 --> 01:24:06,520 INTO THAT I THINK. 1808 01:24:06,520 --> 01:24:07,280 >>OKAY, GREAT. 1809 01:24:07,280 --> 01:24:09,360 AND THEN THE OTHER QUESTION IS I 1810 01:24:09,360 --> 01:24:13,400 WAS TRYING TO UNDERSTAND THE 1811 01:24:13,400 --> 01:24:15,800 SIMULATIONS THAT YOU'RE DOING. 1812 01:24:15,800 --> 01:24:18,320 ARE THE SIMULATIONS EXPLAINING, 1813 01:24:18,320 --> 01:24:23,920 TRYING TO EXPLAIN OXYGEN 1814 01:24:23,920 --> 01:24:25,880 GRADIENTS FROM MITOCHONDRIA INTO 1815 01:24:25,880 --> 01:24:27,400 THE CYTOPLASM? 1816 01:24:27,400 --> 01:24:30,440 I THOUGHT YOUR SIMULATIONS WERE 1817 01:24:30,440 --> 01:24:32,880 FOR OXYGEN GRADIENTS ACROSS 1818 01:24:32,880 --> 01:24:33,080 CELLS. 1819 01:24:33,080 --> 01:24:36,920 >> APOLOGIES FOR THAT. 1820 01:24:36,920 --> 01:24:39,560 SO, I THINK WHAT I WOULD SAY IS 1821 01:24:39,560 --> 01:24:41,600 I'M INTERESTED IN OXYGEN 1822 01:24:41,600 --> 01:24:43,520 GRADIENTS CAUSED BY CELLS WITHIN 1823 01:24:43,520 --> 01:24:47,080 3D STRUCTURES, WHAT HAVE YOU, 1824 01:24:47,080 --> 01:24:48,520 THE PARTICULAR THING WE WERE 1825 01:24:48,520 --> 01:24:50,480 GOING AFTER WITH THE DETAILED 1826 01:24:50,480 --> 01:24:52,120 STRUCTURE OF THE NETWORK WAS 1827 01:24:52,120 --> 01:24:55,200 ACTUALLY EXTENT TO WHICH YOU CAN 1828 01:24:55,200 --> 01:24:59,120 GET OXYGEN GRADIENTS WITHIN A 1829 01:24:59,120 --> 01:24:59,720 CELL. 1830 01:24:59,720 --> 01:25:02,080 SO, IF YOU LOOK AT THE 1831 01:25:02,080 --> 01:25:04,040 MITOCHONDRIAL NETWORK IT'S LIKE 1832 01:25:04,040 --> 01:25:06,920 THIS BASKET AROUND THE NUCLEUS, 1833 01:25:06,920 --> 01:25:08,120 AND IT SPREADS. 1834 01:25:08,120 --> 01:25:11,920 AND SO IF YOU LOOK -- IN THE 1835 01:25:11,920 --> 01:25:13,800 IMAGES OF SINGLE CELLS, THERE'S 1836 01:25:13,800 --> 01:25:16,320 VARIATION IN COLOR, VARIATION IN 1837 01:25:16,320 --> 01:25:17,640 LIFETIME, IN OXYGEN 1838 01:25:17,640 --> 01:25:18,200 CONCENTRATION. 1839 01:25:18,200 --> 01:25:22,320 WE CLEARLY SEE IT. 1840 01:25:22,320 --> 01:25:24,400 AND THERE WERE -- MAYBE WE'RE 1841 01:25:24,400 --> 01:25:25,600 BEARING THE SCARS OF REVIEWS 1842 01:25:25,600 --> 01:25:28,320 WHERE PEOPLE WERE SAYING, NO, 1843 01:25:28,320 --> 01:25:30,320 THESE DON'T HAPPEN. 1844 01:25:30,320 --> 01:25:31,280 THESE OXYGEN GRADIENTS, THIS 1845 01:25:31,280 --> 01:25:32,160 CAN'T HAPPEN. 1846 01:25:32,160 --> 01:25:34,480 AND SO PART OF THE GOAL WITH 1847 01:25:34,480 --> 01:25:35,560 COMPUTATIONAL TOOLS TO SET OUT 1848 01:25:35,560 --> 01:25:37,080 CONDITIONS UNDER WHICH IT CAN 1849 01:25:37,080 --> 01:25:38,400 HAPPEN AND EXPLAIN WHY PEOPLE 1850 01:25:38,400 --> 01:25:39,600 THINK IT MIGHT NOT HAPPEN. 1851 01:25:39,600 --> 01:25:42,640 AND PART OF THE ANSWER THERE IS 1852 01:25:42,640 --> 01:25:44,160 THAT IF YOU HAVE -- I MEAN, IF 1853 01:25:44,160 --> 01:25:47,720 YOU HAVE CELLS THAT ARE GETTING 1854 01:25:47,720 --> 01:25:51,080 OXYGEN IN 3D, THOSE GRADIENTS 1855 01:25:51,080 --> 01:25:52,280 ARE MUCH LESS PRONOUNCED. 1856 01:25:52,280 --> 01:25:54,840 IF YOU'RE UNABLE TO MEASURE AT 1857 01:25:54,840 --> 01:25:58,960 THE SAME SCALE AS 1858 01:25:58,960 --> 01:25:59,960 OXYGEN-CONSUMING STRUCTURES IT'S 1859 01:25:59,960 --> 01:26:02,440 HARDER TO SEE GRADIENTS, BUT NOW 1860 01:26:02,440 --> 01:26:03,240 WE CAN. 1861 01:26:03,240 --> 01:26:05,960 >> GREAT, THANK YOU. 1862 01:26:05,960 --> 01:26:06,240 1863 01:26:06,240 --> 01:26:09,040 >> ANY OTHER QUESTIONS? 1864 01:26:09,040 --> 01:26:15,080 1865 01:26:15,080 --> 01:26:21,200 1866 01:26:21,200 --> 01:26:21,600 THANK YOU. 1867 01:26:21,600 --> 01:26:24,920 >> NEXT IS DR. CHEN, 1868 01:26:24,920 --> 01:26:25,880 CO-DIRECTOR EVER METABOLIC 1869 01:26:25,880 --> 01:26:28,000 CLINICAL RESEARCH UNIT CORES AND 1870 01:26:28,000 --> 01:26:29,200 SUPPORT SERVICES AND SECTION 1871 01:26:29,200 --> 01:26:32,480 CHIEF FOR ENERGY METABOLISM 1872 01:26:32,480 --> 01:26:33,240 SECTION, DIABETES, 1873 01:26:33,240 --> 01:26:36,440 ENDOCRINOLOGY, OBESITY BRANCH. 1874 01:26:36,440 --> 01:26:38,520 AND RESEARCH ON UNDERSTANDING 1875 01:26:38,520 --> 01:26:40,040 HOW HUMAN ENERGY METABOLISM AND 1876 01:26:40,040 --> 01:26:42,320 PHYSICAL ACTIVITY ARE REGULATED 1877 01:26:42,320 --> 01:26:47,040 AND IMPACT DISEASE SUCH AS 1878 01:26:47,040 --> 01:26:47,280 OBESITY. 1879 01:26:47,280 --> 01:26:49,120 >> THANK YOU SO MUCH. 1880 01:26:49,120 --> 01:26:52,960 I DIDN'T KNOW WHAT TO EXPECT 1881 01:26:52,960 --> 01:26:55,880 WITH THE ENCLAVE, CONCLAVE, I 1882 01:26:55,880 --> 01:27:01,920 DIDN'T WEAR MY RED ROBE OR MY 1883 01:27:01,920 --> 01:27:03,200 RED CAP. 1884 01:27:03,200 --> 01:27:08,120 GREAT ARRANGEMENTS OF DR. 1885 01:27:08,120 --> 01:27:10,320 MORGAN, THAT'S WHAT I'M GOING TO 1886 01:27:10,320 --> 01:27:14,720 TALK ABOUT, EXCEPT NOT THE WHOLE 1887 01:27:14,720 --> 01:27:15,360 BODY LEVEL. 1888 01:27:15,360 --> 01:27:19,520 LET'S SEE HOW THIS GOES. 1889 01:27:19,520 --> 01:27:22,680 I APOLOGIZE, THE TITLE, TRYING 1890 01:27:22,680 --> 01:27:24,600 TO PUT BME AND BMR TOGETHER. 1891 01:27:24,600 --> 01:27:33,640 HOW DO WE MEASURE ENERGY 1892 01:27:33,640 --> 01:27:35,600 METABOLISM VIA PRINCIPLES VIA 1893 01:27:35,600 --> 01:27:36,080 BIOMEDICAL ENGINEERING? 1894 01:27:36,080 --> 01:27:38,560 I DO MEASURING OF METABOLIC 1895 01:27:38,560 --> 01:27:40,440 RATE, ONE LETTER DIFFERENCE, 1896 01:27:40,440 --> 01:27:42,920 BME, BMR, SOMETIMES I GET 1897 01:27:42,920 --> 01:27:43,920 CONFUSED WITH THOSE. 1898 01:27:43,920 --> 01:27:46,960 THIS IS A DIFFERENT TALK. 1899 01:27:46,960 --> 01:27:50,240 THERE WILL BE MORE PRESENTED ON 1900 01:27:50,240 --> 01:27:53,160 THE HUMAN RESEARCH AND SO WE DO 1901 01:27:53,160 --> 01:27:55,200 CLINICAL RESEARCH HERE IN THE 1902 01:27:55,200 --> 01:27:56,760 CLINICAL CENTER, MY LAB ON FIFTH 1903 01:27:56,760 --> 01:28:01,360 FLOOR IN THE CLINICAL CENTER. 1904 01:28:01,360 --> 01:28:03,840 SO, AND IT'S BEEN MENTIONED, I'M 1905 01:28:03,840 --> 01:28:05,840 INTERESTED IN BODY WEIGHT 1906 01:28:05,840 --> 01:28:08,320 REGULATION, AND AS IT TURNS OUT 1907 01:28:08,320 --> 01:28:14,240 IT'S A SIMPLE LAW OF 1908 01:28:14,240 --> 01:28:14,880 THERMODYNAMICS, THE ENERGY 1909 01:28:14,880 --> 01:28:16,120 BALANCE, IT'S REALLY THE ENERGY 1910 01:28:16,120 --> 01:28:19,120 IN VERSUS ENERGY OUT. 1911 01:28:19,120 --> 01:28:22,360 WE'VE GOT THE ENERGY IN, WE HAD 1912 01:28:22,360 --> 01:28:23,680 COOKIES AND WHATEVER SNACKS YOU 1913 01:28:23,680 --> 01:28:24,760 HAD, AND THAT WOULD BE THE 1914 01:28:24,760 --> 01:28:25,360 ENERGY IN. 1915 01:28:25,360 --> 01:28:28,480 IF I HAD TO ASK YOU HOW MANY 1916 01:28:28,480 --> 01:28:31,960 CALORIES IS THAT, MOST OF YOU 1917 01:28:31,960 --> 01:28:33,080 DON'T EVEN KNOW. 1918 01:28:33,080 --> 01:28:37,360 IT DEPENDS ON WHAT YOU ATE AND 1919 01:28:37,360 --> 01:28:40,720 HOW MUCH IS DIGESTED. 1920 01:28:40,720 --> 01:28:42,040 FORTUNATELY I CAN MEASURE ENERGY 1921 01:28:42,040 --> 01:28:43,560 OUT MORE PRECISELY. 1922 01:28:43,560 --> 01:28:45,680 THAT'S WHAT WE FOCUS ON. 1923 01:28:45,680 --> 01:28:52,680 WE CAN MEASURE BODY COMPOSITION, 1924 01:28:52,680 --> 01:28:54,080 THAT'S ANOTHER BIG PART OF MY 1925 01:28:54,080 --> 01:28:55,160 LAB, MEASURE OF BODY 1926 01:28:55,160 --> 01:28:55,840 COMPOSITION. 1927 01:28:55,840 --> 01:28:57,560 IF YOU HAVE TWO OF THE THREE 1928 01:28:57,560 --> 01:28:58,560 COMPONENTS YOU CAN DETERMINE 1929 01:28:58,560 --> 01:28:58,840 ENERGY IN. 1930 01:28:58,840 --> 01:29:01,160 THIS IS WHAT WE'RE GOING TO TALK 1931 01:29:01,160 --> 01:29:01,880 ABOUT. 1932 01:29:01,880 --> 01:29:03,720 PARTICULARLY DIFFERENT 1933 01:29:03,720 --> 01:29:05,880 COMPONENTS OF ENERGY 1934 01:29:05,880 --> 01:29:07,520 EXPENDITURE, I'LL SHRINK DOWN TO 1935 01:29:07,520 --> 01:29:08,760 BMR, BUT THERE ARE OTHER 1936 01:29:08,760 --> 01:29:18,280 COMPONENTS WE ALSO TRY TO 1937 01:29:18,280 --> 01:29:18,880 MEASURE. 1938 01:29:18,880 --> 01:29:21,480 AS HISTORY GOES THE CONCEPT OF 1939 01:29:21,480 --> 01:29:23,920 HEAT HAS BEEN REALLY PROPOSED BY 1940 01:29:23,920 --> 01:29:26,600 ARISTOTLE, THE FOOD MUST BE 1941 01:29:26,600 --> 01:29:30,960 CAPABLE OF BEING DIGESTED AND 1942 01:29:30,960 --> 01:29:35,520 THOSE ACTIVITIES OF DIGESTION, 1943 01:29:35,520 --> 01:29:37,760 HEAT, DIRECT CALORIEMETRY, YOU 1944 01:29:37,760 --> 01:29:39,600 MEASURE THE HEAT, RIGHT? 1945 01:29:39,600 --> 01:29:46,720 BUT THE -- I KEEP GOING TO THE 1946 01:29:46,720 --> 01:29:47,840 WRONG ONE. 1947 01:29:47,840 --> 01:29:58,240 SO, 1770s IN PARIS, THIS 1948 01:29:58,920 --> 01:29:59,840 INVESTIGATOR LAVASIER MEASURED 1949 01:29:59,840 --> 01:30:01,800 BY RABBIT GENERATING HEAT AND 1950 01:30:01,800 --> 01:30:06,400 ICE BATH IN THE PREVIOUS SLIDE, 1951 01:30:06,400 --> 01:30:09,680 BUT HE DISCOVERED, HE AND HIS 1952 01:30:09,680 --> 01:30:11,360 PARTNER DISCOVERED IF YOU DON'T 1953 01:30:11,360 --> 01:30:13,880 FEED AIR INTO THIS SYSTEM, THE 1954 01:30:13,880 --> 01:30:15,520 RABBIT DIES. 1955 01:30:15,520 --> 01:30:17,600 THE HEAT DOESN'T MATTER OR NOT. 1956 01:30:17,600 --> 01:30:21,440 IN HUMANS, YOU START TO FOCUS ON 1957 01:30:21,440 --> 01:30:23,280 MEASURING GASES EXCHANGES, AND 1958 01:30:23,280 --> 01:30:24,920 THERE BY MEASURING IN THIS 1959 01:30:24,920 --> 01:30:27,560 UNITS, WHICH I LOVE THE MOST, 1960 01:30:27,560 --> 01:30:29,840 THE PUBLISHED AND DOCUMENTED BY 1961 01:30:29,840 --> 01:30:36,400 SCIENCE AND NUTRITION GRAHAM 1962 01:30:36,400 --> 01:30:38,920 LUSK, RESTING MAN REQUIRED 1200 1963 01:30:38,920 --> 01:30:44,720 AT 26 DEGREES C, 1400 AT 12 1964 01:30:44,720 --> 01:30:47,920 DEGREES C, WITH DIFFERENCE IN 1965 01:30:47,920 --> 01:30:48,800 METABOLISM CONSUMPTION. 1966 01:30:48,800 --> 01:30:53,200 ADVANCE 100 YEARS, A GERMAN 1967 01:30:53,200 --> 01:30:54,840 SCIENTIST DEVISED A ROOM WHICH 1968 01:30:54,840 --> 01:30:59,000 WE CALLED THE ROOM CALORIEMETER, 1969 01:30:59,000 --> 01:31:00,720 REGULATE TEMPERATURE BY OPENING 1970 01:31:00,720 --> 01:31:03,480 WINDOWS OR DOORS, SEALED IN 1971 01:31:03,480 --> 01:31:04,880 ONE-WAY FLOW MATTER, OPEN 1972 01:31:04,880 --> 01:31:06,520 CIRCUIT MATTER, WHICH HE 1973 01:31:06,520 --> 01:31:07,840 MEASURED THE CO2 PRODUCTION, SO 1974 01:31:07,840 --> 01:31:10,360 NOW YOU HAVE OXYGEN CONSUMPTION 1975 01:31:10,360 --> 01:31:10,560 AND CO2. 1976 01:31:10,560 --> 01:31:11,520 WHY ARE THOSE IMPORTANT? 1977 01:31:11,520 --> 01:31:15,800 I'LL SHOW YOU IN THE NEXT SLIDE 1978 01:31:15,800 --> 01:31:17,680 OR SO. 1979 01:31:17,680 --> 01:31:24,040 SO, ANOTHER 100 YEARS LATER, 1980 01:31:24,040 --> 01:31:28,720 1900s, METABOLIC CHAMBER CAME 1981 01:31:28,720 --> 01:31:30,560 IN, BUT IT IS -- THE REASON IT'S 1982 01:31:30,560 --> 01:31:34,440 A CHAMBER, BUILT IN THE BASEMENT 1983 01:31:34,440 --> 01:31:42,960 OF A UNIVERSITY BUILDING, METAL 1984 01:31:42,960 --> 01:31:45,760 ROOM, COLD AND DARK, THEY CALLED 1985 01:31:45,760 --> 01:31:48,160 IT THE CHAMBER. 1986 01:31:48,160 --> 01:31:50,640 THE FIRST OF ITS KIND MEASURING 1987 01:31:50,640 --> 01:31:52,480 HUMANS THAT DOES TWO PARTS. 1988 01:31:52,480 --> 01:31:57,920 ONE IS DIRECT, OR HEAT 1989 01:31:57,920 --> 01:31:59,360 MEASUREMENT, BY INSULATING WITH 1990 01:31:59,360 --> 01:32:00,640 COPPER WATER AND WATER 1991 01:32:00,640 --> 01:32:01,600 CIRCULATING THROUGH IT, MEASURE 1992 01:32:01,600 --> 01:32:04,160 THE TEMPERATURE CHANGE OF THE 1993 01:32:04,160 --> 01:32:06,480 WATER, THE VOLUME AND THE 1994 01:32:06,480 --> 01:32:07,560 CONDUCTIVITY OF THE WATER, 1995 01:32:07,560 --> 01:32:10,920 MEASURES THE DIRECT HEAT 1996 01:32:10,920 --> 01:32:12,040 EXCHANGE. 1997 01:32:12,040 --> 01:32:14,000 SIMULTANEOUSLY THEY MEASURE GAS 1998 01:32:14,000 --> 01:32:16,320 EXCHANGE, BY CIRCULATING AIR AND 1999 01:32:16,320 --> 01:32:23,200 MEASURE THE CO2 PRODUCTION BY 2000 01:32:23,200 --> 01:32:25,160 THE ABSORPTION, SODA LIME, 2001 01:32:25,160 --> 01:32:28,880 SULFURIC ACID, YOU CAN MEASURE 2002 01:32:28,880 --> 01:32:30,280 BOTH DIRECT, INDIRECT, HEAT 2003 01:32:30,280 --> 01:32:32,600 EXCHANGE, DIRECTLY BY THE HEAT, 2004 01:32:32,600 --> 01:32:35,120 AND INDIRECTLY BY GAS EXCHANGE. 2005 01:32:35,120 --> 01:32:45,680 AND THUS BY VALIDATED PRINCIPAL 2006 01:32:56,120 --> 01:32:56,680 OF INDIRECT CALORIEMETRY, WHEN 2007 01:32:56,680 --> 01:33:00,000 YOU HAVE A MOLECULE OF FAT YOU 2008 01:33:00,000 --> 01:33:03,600 NEED MORE OXYGEN THAN YOU 2009 01:33:03,600 --> 01:33:06,760 PRODUCE IN CO2. 2010 01:33:06,760 --> 01:33:07,240 PROTEIN, MIXED PROTEIN, 2011 01:33:07,240 --> 01:33:11,240 DIFFERENT EXCHANGE. 2012 01:33:11,240 --> 01:33:14,520 BY THE DIFFERENT EXCHANGES OF 2013 01:33:14,520 --> 01:33:15,920 CARBOHYDRATE, FAT, PROTEIN, 2014 01:33:15,920 --> 01:33:18,680 MACRONUTRIENTS, SO WHEN YOU EAT 2015 01:33:18,680 --> 01:33:19,280 THE COOKIES, CARBOHYDRATES AND 2016 01:33:19,280 --> 01:33:23,280 FAT WITH A LITTLE BIT OF 2017 01:33:23,280 --> 01:33:24,600 PROTEIN, REMEMBER YOUR LUNCH, 2018 01:33:24,600 --> 01:33:30,840 THAT'S THE THINGS WE ALSO BURN. 2019 01:33:30,840 --> 01:33:33,280 WE AS AN ENGINE, FLEX FUEL 2020 01:33:33,280 --> 01:33:37,080 BURNER, AS WE DO, SO WE MEASURE 2021 01:33:37,080 --> 01:33:40,040 THESE THREE COMPONENTS, NAMELY 2022 01:33:40,040 --> 01:33:41,800 CO2 PRODUCTION, OXYGEN 2023 01:33:41,800 --> 01:33:44,040 CONSUMPTION, AND URINARY 2024 01:33:44,040 --> 01:33:44,880 NITROGEN PRODUCTION, PROTEIN 2025 01:33:44,880 --> 01:33:46,640 TURNOVER, WE CAN CALCULATE 2026 01:33:46,640 --> 01:33:46,920 EVERYTHING. 2027 01:33:46,920 --> 01:33:52,000 THAT'S EXACTLY WHAT WE DO 2028 01:33:52,000 --> 01:33:52,520 INDIRECTLY. 2029 01:33:52,520 --> 01:33:53,520 WE HAVE ANOTHER REPRESENTATION 2030 01:33:53,520 --> 01:33:56,080 OF THE WHAT FUEL WE'RE BURNING, 2031 01:33:56,080 --> 01:33:59,320 THE RATIO OF OXYGEN CONSUMPTION 2032 01:33:59,320 --> 01:34:03,600 AND CO2, SIMPLE RATIO OF CO2 2033 01:34:03,600 --> 01:34:05,480 PROSECUTION OVER OXYGEN 2034 01:34:05,480 --> 01:34:15,000 CONSUMPTION, IF IT'S ONE, 2035 01:34:15,000 --> 01:34:19,880 CARBOHYDRATE, 6: 6, WESTERN 2036 01:34:19,880 --> 01:34:23,680 DIET IS 0.85. 2037 01:34:23,680 --> 01:34:25,200 HIGHER THAN 1 IS LIPOGENESIS, 2038 01:34:25,200 --> 01:34:29,680 BELOW 7 WHICH MOST PEOPLE DON'T, 2039 01:34:29,680 --> 01:34:30,560 ALCOHOL OXIDATION PROCESS, 2040 01:34:30,560 --> 01:34:34,040 SOMETIMES WE BURN ALCOHOL AS A 2041 01:34:34,040 --> 01:34:34,600 VALIDATION PROCESS. 2042 01:34:34,600 --> 01:34:38,000 SO THEN YOU HAVE THE STANDARD 2043 01:34:38,000 --> 01:34:40,320 EQUATION, DEVELOPED 100 YEARS 2044 01:34:40,320 --> 01:34:41,400 AGO. 2045 01:34:41,400 --> 01:34:43,400 THAT YOU CAN ESSENTIALLY 2046 01:34:43,400 --> 01:34:49,600 CALCULATE THE ENERGY EXPENDITURE 2047 01:34:49,600 --> 01:34:52,680 OR METABOLIC RATE, A MINUTE, 2048 01:34:52,680 --> 01:34:56,080 HOUR, DAY, CALCULATED BY THIS 2049 01:34:56,080 --> 01:34:57,920 EQUATION, OXYGEN CONSUMPTION, 2050 01:34:57,920 --> 01:34:58,720 CO2 PRODUCTION. 2051 01:34:58,720 --> 01:34:59,920 ALL THAT'S BEING SAID, HOW DOES 2052 01:34:59,920 --> 01:35:01,960 IT DO IT? 2053 01:35:01,960 --> 01:35:04,960 WELL, WE HAVE TO CALCULATE AN 2054 01:35:04,960 --> 01:35:08,760 OPEN CIRCUIT MATTER, INCORRECT 2055 01:35:08,760 --> 01:35:11,640 CALORIEMETRY, WHAT IS OXYGEN 2056 01:35:11,640 --> 01:35:14,160 CONSUMPTION, VOLUME OF INSPIRED 2057 01:35:14,160 --> 01:35:15,440 OXYGEN AND FRACTIONAL 2058 01:35:15,440 --> 01:35:17,320 CONCENTRATION OF OXYGEN TIMES 2059 01:35:17,320 --> 01:35:22,560 VOLUME OF WHAT'S INSPIRED, MINUS 2060 01:35:22,560 --> 01:35:25,400 THE EXPIRED OXYGEN, SORT OF 2061 01:35:25,400 --> 01:35:25,840 CONTENT. 2062 01:35:25,840 --> 01:35:28,680 SIMILARLY YOU DO THIS WITH CO2. 2063 01:35:28,680 --> 01:35:32,120 SO, THIS IS THE SIMPLE EQUATION, 2064 01:35:32,120 --> 01:35:33,840 BEING BIOMEDICAL ENGINEERS WE 2065 01:35:33,840 --> 01:35:35,120 HAVE TO SHOW EQUATIONS, RIGHT? 2066 01:35:35,120 --> 01:35:37,680 THIS IS THE SCHEMATIC OF HOW 2067 01:35:37,680 --> 01:35:38,320 IT'S DONE. 2068 01:35:38,320 --> 01:35:39,840 ANOTHER 100 YEARS, NOT BY 2069 01:35:39,840 --> 01:35:43,000 CHANCE, BUT JUST THE FACT THESE 2070 01:35:43,000 --> 01:35:50,040 THINGS ARE ADVANCED IN 100-YEAR 2071 01:35:50,040 --> 01:35:50,280 CYCLES. 2072 01:35:50,280 --> 01:35:52,760 THIS IS A METABOLIC CHAMBER 2073 01:35:52,760 --> 01:35:53,640 BUILT IN SWITZERLAND, SINCE 2074 01:35:53,640 --> 01:35:57,480 MOVED TO THE U.S., WITH CHAMBERS 2075 01:35:57,480 --> 01:36:03,720 IN THE RESEARCH FACILITIES AND 2076 01:36:03,720 --> 01:36:06,240 NIDDK, PHOENIX, AND PENNINGTON 2077 01:36:06,240 --> 01:36:07,120 BIORESEARCH CENTER, INDIRECT 2078 01:36:07,120 --> 01:36:08,880 MEASUREMENT OF OXYGEN AND CO2 2079 01:36:08,880 --> 01:36:11,560 NOW, YOU CAN SEE SMALL AIR ROOM 2080 01:36:11,560 --> 01:36:19,040 AT THE TIME 17 CUBIC METERS, 2081 01:36:19,040 --> 01:36:21,720 DESK, SINK, TOILET, A BED ON THE 2082 01:36:21,720 --> 01:36:24,440 SIDE OR SOFA, THEY HAVE A RADAR 2083 01:36:24,440 --> 01:36:26,280 SYSTEM TO MEASURE MOVEMENT, AIR 2084 01:36:26,280 --> 01:36:28,240 LOCKED, FOOD IN, WHAT NOT, I'LL 2085 01:36:28,240 --> 01:36:29,880 SHOW YOU THE REAL PICTURE. 2086 01:36:29,880 --> 01:36:32,200 AND THEN THEY DRAW THE AIR OUT 2087 01:36:32,200 --> 01:36:35,800 AND SAMPLE THE FLOW RATE WHICH 2088 01:36:35,800 --> 01:36:37,320 HAS STATIC VOLUME AND DYNAMIC 2089 01:36:37,320 --> 01:36:41,920 VOLUME, WHICH IS OPEN CIRCUIT 2090 01:36:41,920 --> 01:36:46,400 DESIGN NOW, AIR CONDITIONING AND 2091 01:36:46,400 --> 01:36:47,960 FILTERING, MOISTURE, MEASURE 2092 01:36:47,960 --> 01:36:51,000 OXYGEN BY PARAMAGNETIC SENSOR, 2093 01:36:51,000 --> 01:36:59,680 CONTINUOUS SENSOR AND CO2 BY 2094 01:36:59,680 --> 01:37:00,640 NEAR-INFRARED SENSOR, MODERNIZED 2095 01:37:00,640 --> 01:37:01,640 CALORIEMETRY BY ITSELF. 2096 01:37:01,640 --> 01:37:03,920 THIS IS THE CHAMBER I BUILT IN 2097 01:37:03,920 --> 01:37:05,880 THE CLINICAL CENTER. 2098 01:37:05,880 --> 01:37:07,080 WE RECRUITED FROM VANDERBILT, 2099 01:37:07,080 --> 01:37:12,920 WE, MY POSTDOC AND I, CAME HERE 2100 01:37:12,920 --> 01:37:14,760 2006, BUILT THREE CHAMBERS, 2101 01:37:14,760 --> 01:37:15,560 7th FLOOR CLINICAL CENTER, 2102 01:37:15,560 --> 01:37:18,840 THEY STILL OPERATE MOST OF THE 2103 01:37:18,840 --> 01:37:19,040 TIME. 2104 01:37:19,040 --> 01:37:22,280 SO IT'S A VOLUME OF 27,000 2105 01:37:22,280 --> 01:37:23,680 LITERS NOW. 2106 01:37:23,680 --> 01:37:25,520 APPLY DR. MORGAN'S PRINCIPLE, 2107 01:37:25,520 --> 01:37:27,960 THINK ABOUT 21% OF THE AMBIENT 2108 01:37:27,960 --> 01:37:31,360 AIR IS OXYGEN, AND SO WE MEASURE 2109 01:37:31,360 --> 01:37:34,720 DYNAMIC RANGE OF 1%, 20 TO 21% 2110 01:37:34,720 --> 01:37:36,480 OXYGEN, 0 TO 1% CO2. 2111 01:37:36,480 --> 01:37:40,720 YOU CAN DO THE MATH, BUT IF IT'S 2112 01:37:40,720 --> 01:37:42,720 1/5 OF THE AIR IS OXYGEN, YOU 2113 01:37:42,720 --> 01:37:45,520 END UP WITH MAYBE 70,000 LITERS 2114 01:37:45,520 --> 01:37:49,960 OF OXYGEN, AND BUT WE MEASURE 2115 01:37:49,960 --> 01:37:52,560 OXYGEN CONSUMPTION RATE PER 2116 01:37:52,560 --> 01:37:54,400 MINUTE BETWEEN 200 AND 300 2117 01:37:54,400 --> 01:37:55,080 MILLILITERS, REALLY THE 2118 01:37:55,080 --> 01:37:56,280 PRECISION MEASUREMENT OF THESE 2119 01:37:56,280 --> 01:37:57,240 THINGS THAT MATTERS. 2120 01:37:57,240 --> 01:37:59,120 TURNS OUT THERE ARE ONLY A FEW 2121 01:37:59,120 --> 01:38:01,200 OF THESE FACILITIES AROUND THE 2122 01:38:01,200 --> 01:38:01,400 WORLD. 2123 01:38:01,400 --> 01:38:04,200 AND THIS IS SOME OF THE PICTURES 2124 01:38:04,200 --> 01:38:05,600 SHOWING DIFFERENT DESIGNS, BY 2125 01:38:05,600 --> 01:38:10,520 AND LARGE SIMILAR IN VOLUME, 2126 01:38:10,520 --> 01:38:13,120 SMALLEST IS PROBABLY A 2127 01:38:13,120 --> 01:38:16,440 UNIVERSITY CHAMBER, 17 CUBIC 2128 01:38:16,440 --> 01:38:22,880 METERS, OURS IS 27, ANSCHUTZ IS 2129 01:38:22,880 --> 01:38:26,240 25 CUBIC MEETERS AND PENNINGTON 2130 01:38:26,240 --> 01:38:28,440 30 CUBIC METERS, FOUR CHAMBERS, 2131 01:38:28,440 --> 01:38:29,880 FACILITIES AROUND THE WORLD, WE 2132 01:38:29,880 --> 01:38:31,560 KEEP TRACK OF THEM SO WE JUST 2133 01:38:31,560 --> 01:38:34,240 HAD A MEETING AND FIGURED OUT 2134 01:38:34,240 --> 01:38:35,360 HOW MANY METABOLIC CHAMBERS, WE 2135 01:38:35,360 --> 01:38:39,480 NEED TO UPDATE BECAUSE CHINA 2136 01:38:39,480 --> 01:38:41,160 ADDED FOUR CHAMBERS IN THE 2137 01:38:41,160 --> 01:38:42,240 PANDEMIC ERA. 2138 01:38:42,240 --> 01:38:44,640 AND WE LOST TWO CHAMBERS 2139 01:38:44,640 --> 01:38:46,640 INCLUDING MY OLD CHAMBERS IN 2140 01:38:46,640 --> 01:38:47,840 VANDERBILT IN NASHVILLE, 2141 01:38:47,840 --> 01:38:48,240 TENNESSEE. 2142 01:38:48,240 --> 01:38:50,520 SO, THERE'S SOME UPDATES ON 2143 01:38:50,520 --> 01:38:52,080 THESE THINGS BUT ROUGHLY 40 OF 2144 01:38:52,080 --> 01:38:53,840 THESE FACILITIES AROUND THE 2145 01:38:53,840 --> 01:38:56,240 WORLD, THAT CAN DO THESE 2146 01:38:56,240 --> 01:38:59,320 METABOLIC CHAMBER MEASUREMENTS. 2147 01:38:59,320 --> 01:38:59,680 HUMAN CHAMBERS. 2148 01:38:59,680 --> 01:39:02,920 WITH ANY CHAMBER WE TALK ABOUT 2149 01:39:02,920 --> 01:39:04,680 TESTS, REPRODUCIBILITY, WE DID 2150 01:39:04,680 --> 01:39:11,040 THE STUDY WHEN WE FIRST HAD THE 2151 01:39:11,040 --> 01:39:13,040 FACILITIES RUNNING, 10 SUBJECTS 2152 01:39:13,040 --> 01:39:21,280 IN THE CHAMBER, THIS IS WHEN IRB 2153 01:39:21,280 --> 01:39:21,960 ALLOWED INVESTIGATOR ITSELF IN 2154 01:39:21,960 --> 01:39:23,160 THE INVESTIGATION. 2155 01:39:23,160 --> 01:39:26,880 I'M HERE SOMEWHERE. 2156 01:39:26,880 --> 01:39:30,240 THIS IS A BLUNT ALMOND PLOT, 24 2157 01:39:30,240 --> 01:39:33,680 HOURS, VERSUS AVERAGE OF THE 2158 01:39:33,680 --> 01:39:33,840 TWO. 2159 01:39:33,840 --> 01:39:38,880 YOU CAN SEE THE CURVE IS PRETTY 2160 01:39:38,880 --> 01:39:41,560 FLAT, MEANING THERE'S REALLY NOT 2161 01:39:41,560 --> 01:39:42,680 MUCH BIAS, DIFFERENCE ACCORDING 2162 01:39:42,680 --> 01:39:43,600 TO SIGNAL. 2163 01:39:43,600 --> 01:39:49,400 AND WE CAN ALSO DO THAT WITH THE 2164 01:39:49,400 --> 01:39:53,440 RQ, RESPIRATORY QUOTIENT, .85 IS 2165 01:39:53,440 --> 01:39:55,320 AVERAGE OF RQ OF THE WHOLE DAY. 2166 01:39:55,320 --> 01:39:56,880 THERE'S A VARIATION. 2167 01:39:56,880 --> 01:39:59,800 SOME PEOPLE CLOSE TO .8, SOME 2168 01:39:59,800 --> 01:40:03,000 ACTUALLY CLOSE TO .9. 2169 01:40:03,000 --> 01:40:11,320 BUT THE ENERGY EXPENDITURE, 2170 01:40:11,320 --> 01:40:12,400 KILOCALORIES TODAY, 1500 TO 2171 01:40:12,400 --> 01:40:14,160 4,000 PER DAY, BODY SIZE 2172 01:40:14,160 --> 01:40:15,120 MATTERS, WHICH I'LL SHOW ANOTHER 2173 01:40:15,120 --> 01:40:18,080 SLIDE OF THAT. 2174 01:40:18,080 --> 01:40:19,400 SO WHEN WE LOOK WHAT'S IMPORTANT 2175 01:40:19,400 --> 01:40:22,680 IN THE SLIDE, THIS NUMBER RIGHT 2176 01:40:22,680 --> 01:40:24,080 HERE, STANDARD DEVIATION, PLUS 2177 01:40:24,080 --> 01:40:25,400 OR MINUS 63. 2178 01:40:25,400 --> 01:40:27,920 THE DIFFERENCE OBVIOUSLY IS ALSO 2179 01:40:27,920 --> 01:40:29,800 IMPORTANT, 50 KCALS PER DAY, 2180 01:40:29,800 --> 01:40:33,720 BETWEEN DAY 1 AND DAY 2, WE'LL 2181 01:40:33,720 --> 01:40:35,720 CALL IT FIRST DAY EFFECT, 2182 01:40:35,720 --> 01:40:36,880 DIFFERENT ENVIRONMENT, FIGURE 2183 01:40:36,880 --> 01:40:40,600 OUT WHERE TO PUT YOUR CHOOSE, 2184 01:40:40,600 --> 01:40:42,360 LAPTOPS, BECAUSE IT'S INDIRECT 2185 01:40:42,360 --> 01:40:43,800 MEASUREMENT, YOU CAN BRING LAP 2186 01:40:43,800 --> 01:40:45,400 TOPS AND WHAT NOT BECAUSE WE'RE 2187 01:40:45,400 --> 01:40:46,960 NOT MEASURING HEAT, WE'RE 2188 01:40:46,960 --> 01:40:48,480 MEASURING GASES. 2189 01:40:48,480 --> 01:40:51,320 YOU SEE THE CHAMBER, THERE'S NO 2190 01:40:51,320 --> 01:40:54,080 PLANTS, NO THINGS THAT CONSUME 2191 01:40:54,080 --> 01:40:56,800 OXYGEN OR PRODUCE OXYGEN. 2192 01:40:56,800 --> 01:40:59,080 SO, NEVERTHELESS WE CAN ACTUALLY 2193 01:40:59,080 --> 01:41:01,920 HAVE A REPRODUCIBILITY OF ABOUT 2194 01:41:01,920 --> 01:41:04,000 3% OF THE 24 HOUR ENERGY 2195 01:41:04,000 --> 01:41:05,120 EXPENDITURE, PROBABLY AS HIGH AS 2196 01:41:05,120 --> 01:41:07,840 IT'S GOING TO GO. 2197 01:41:07,840 --> 01:41:12,680 AND RQ IS ALSO 2%, AND PLUS OR 2198 01:41:12,680 --> 01:41:13,400 MINUS 2%. 2199 01:41:13,400 --> 01:41:17,480 SO LET'S TAKE A LOOK AT VERY 2200 01:41:17,480 --> 01:41:19,360 QUICK WHAT INDIVIDUAL DATA LOOKS 2201 01:41:19,360 --> 01:41:20,880 LIKE, TAKING TWO DATA POINTS, 2202 01:41:20,880 --> 01:41:22,280 ONE WITH HIGHEST ENERGY 2203 01:41:22,280 --> 01:41:24,480 EXPENDITURE AND ONE WITH LOWEST, 2204 01:41:24,480 --> 01:41:26,360 HERE IS THE LOWEST. 2205 01:41:26,360 --> 01:41:30,640 THESE ARE NOW EXPRESSED IN A 2206 01:41:30,640 --> 01:41:32,840 24-HOUR MINUTE-BY-MINUTE CYCLE, 2207 01:41:32,840 --> 01:41:34,040 THIS IS MINUTE-BY-MINUTE CHANGES 2208 01:41:34,040 --> 01:41:37,120 IN KCALS PER MINUTE NOW. 2209 01:41:37,120 --> 01:41:39,080 SO SLEEPING, THIS LEAN HEALTHY 2210 01:41:39,080 --> 01:41:40,360 FEMALE SUBJECT IS BURNING ABOUT 2211 01:41:40,360 --> 01:41:42,240 A LITTLE LESS THAN 1 KCAL PER 2212 01:41:42,240 --> 01:41:45,600 MINUTE, THIS IS HOW EFFICIENT WE 2213 01:41:45,600 --> 01:41:47,360 ARE, 200 MILLILITERS OF OXYGEN 2214 01:41:47,360 --> 01:41:48,440 USED PER MINUTE. 2215 01:41:48,440 --> 01:41:53,480 WE WANT TO MEASURE WITHIN 5% OF 2216 01:41:53,480 --> 01:41:53,800 THAT. 2217 01:41:53,800 --> 01:41:54,960 TALK ABOUT PRECISION, SQUEEZING 2218 01:41:54,960 --> 01:41:56,200 ANY SIGNAL OUT OF THAT. 2219 01:41:56,200 --> 01:41:58,640 AND THE BLUE AND BLACK LINE 2220 01:41:58,640 --> 01:42:00,160 SHOWS REPRODUCIBILITY OF DAY 1 2221 01:42:00,160 --> 01:42:02,560 AND DAY 2, AND THAT'S TOTAL OF 2222 01:42:02,560 --> 01:42:02,800 1%. 2223 01:42:02,800 --> 01:42:04,960 YOU'LL SEE THERE'S SOME BUMPS 2224 01:42:04,960 --> 01:42:07,080 DURING THE DAY, AND IT'S NOT 2225 01:42:07,080 --> 01:42:09,480 EXERCISE AT ALL, SO WE HAVE 2226 01:42:09,480 --> 01:42:11,120 MOTION SENSOR IN THERE, THESE 2227 01:42:11,120 --> 01:42:12,760 ARE PURELY DUE TO MEAL WHICH 2228 01:42:12,760 --> 01:42:16,120 I'LL TELL YOU ABOUT MEAL INDUCED 2229 01:42:16,120 --> 01:42:20,080 OR THERMO EFFECT OF FEEDING. 2230 01:42:20,080 --> 01:42:21,280 THE DYNAMIC RANGE COMES IN, 2231 01:42:21,280 --> 01:42:22,960 DIFFERENT SUBJECT LIKES TO RUN. 2232 01:42:22,960 --> 01:42:25,200 WE HAD A TREADMILL IN THE ROOM, 2233 01:42:25,200 --> 01:42:28,520 YOU CAN SEE IT HERE. 2234 01:42:28,520 --> 01:42:31,040 HE WOULD RUN 10 MILES PER HOUR, 2235 01:42:31,040 --> 01:42:31,680 30 MINUTES IN THE MORNING, 30 2236 01:42:31,680 --> 01:42:33,440 MINUTES IN THE AFTERNOON. 2237 01:42:33,440 --> 01:42:36,200 THERE BY SORT OF DEMONSTRATING 2238 01:42:36,200 --> 01:42:38,480 THE DYNAMIC RANGE OF MOST 2239 01:42:38,480 --> 01:42:40,000 INDIVIDUALS CAN RANGE FROM 10 2240 01:42:40,000 --> 01:42:42,280 TIMES OF THE BASIC METABOLIC 2241 01:42:42,280 --> 01:42:42,480 RATE. 2242 01:42:42,480 --> 01:42:44,600 IT DOESN'T LAST VERY LONG IN HIS 2243 01:42:44,600 --> 01:42:46,320 CASE AN HOUR, BUT MOST SUBJECTS 2244 01:42:46,320 --> 01:42:48,960 CAN'T DO THAT, RIGHT? 2245 01:42:48,960 --> 01:42:52,560 SO WHAT IT DEMONSTRATES IS NOT 2246 01:42:52,560 --> 01:42:54,320 ONLY REPRODUCIBILITY, 2247 01:42:54,320 --> 01:42:54,960 MINUTE-BY-MINUTE LEVEL, BUT ALSO 2248 01:42:54,960 --> 01:42:58,480 SHOWS WE CAN TEASE OUT 2249 01:42:58,480 --> 01:43:00,480 COMPONENTS, SUCH AS EXERCISE, 2250 01:43:00,480 --> 01:43:03,520 SLEEP, SUCH AS DIFFERENT MEALS, 2251 01:43:03,520 --> 01:43:03,840 RIGHT? 2252 01:43:03,840 --> 01:43:06,680 AND SO HOW DO WE DO THAT? 2253 01:43:06,680 --> 01:43:08,760 YOU KNOW, YOU CAN DRAW THE 2254 01:43:08,760 --> 01:43:13,920 LINES, THE WAY WE DO THAT IS 2255 01:43:13,920 --> 01:43:16,320 CORRELATE THE ENERGY EXPENDITURE 2256 01:43:16,320 --> 01:43:17,840 IN MINUTE-BY-MINUTE FASHION 2257 01:43:17,840 --> 01:43:19,920 VERSUS ACTIVITY LEVEL, EITHER BY 2258 01:43:19,920 --> 01:43:22,320 SORT OF PORTABLE OR WEARABLE 2259 01:43:22,320 --> 01:43:23,120 SENSORS, OR BY RADAR SYSTEM 2260 01:43:23,120 --> 01:43:25,520 WHICH WE PUT IN THERE, THERE ARE 2261 01:43:25,520 --> 01:43:28,480 TWO SMALL BLACK DOTS, YOU CAN 2262 01:43:28,480 --> 01:43:30,440 PROBABLY SEE ON TOP OF THE 2263 01:43:30,440 --> 01:43:32,720 CEILING, THESE ARE MOTION 2264 01:43:32,720 --> 01:43:35,240 SENSORS, MUSEUM GRADE MOTION 2265 01:43:35,240 --> 01:43:37,440 SENSORS DETECTED, SO THAT'S WHAT 2266 01:43:37,440 --> 01:43:38,200 WE USE. 2267 01:43:38,200 --> 01:43:40,400 ENGINEERS, WE WIRE THEM, RIGHT? 2268 01:43:40,400 --> 01:43:42,800 AND WE MEASURE THE ACTIVITY. 2269 01:43:42,800 --> 01:43:45,880 SO WITH THE ACTIVITY INCREASES 2270 01:43:45,880 --> 01:43:48,040 YOU'LL SEE THAT ACTIVITY GOES 2271 01:43:48,040 --> 01:43:49,360 UP, ENERGY EXPENDITURE GOES UP 2272 01:43:49,360 --> 01:43:53,080 DUE TO ACTIVITY RELATED TO THE 2273 01:43:53,080 --> 01:43:54,000 MOVEMENT. 2274 01:43:54,000 --> 01:43:56,800 THEN YOU DRAW THIS INTERCEPT, 2275 01:43:56,800 --> 01:43:58,560 ZERO INTERCEPT, ZERO MOVEMENT, 2276 01:43:58,560 --> 01:44:01,080 THAT INTERCEPT IS HIGHER THAN 2277 01:44:01,080 --> 01:44:02,280 MEASURED BASIC METABOLIC, I SHOW 2278 01:44:02,280 --> 01:44:05,880 YOU HOW TO MEASURE THAT. 2279 01:44:05,880 --> 01:44:07,640 THE DIFFERENCE, THERMIC EFFECT 2280 01:44:07,640 --> 01:44:08,880 OF FEEDING, CLASSIFY ALL 2281 01:44:08,880 --> 01:44:09,720 MEASUREMENTS OF COMPONENTS WHICH 2282 01:44:09,720 --> 01:44:13,880 THESE ARE THREE MAJOR COMPONENTS 2283 01:44:13,880 --> 01:44:17,080 OF TOTAL ENERGY EXPENDITURE, 2284 01:44:17,080 --> 01:44:18,080 RESIDENT ENERGY EXPENDITURE, 2285 01:44:18,080 --> 01:44:21,360 THERMIC EFFECT OF FEEDING AND 2286 01:44:21,360 --> 01:44:22,880 YOUR ACTIVITY ENERGY 2287 01:44:22,880 --> 01:44:23,720 EXPENDITURE. 2288 01:44:23,720 --> 01:44:23,920 OKAY. 2289 01:44:23,920 --> 01:44:26,560 SO IT TURNS OUT IN OUR WESTERN 2290 01:44:26,560 --> 01:44:29,200 LIFESTYLE THE RESTING ENERGY 2291 01:44:29,200 --> 01:44:30,640 EXPENDITURE OR BASIC METABOLIC 2292 01:44:30,640 --> 01:44:32,920 EXPLAINED 60 TO 75% OF A TOTAL 2293 01:44:32,920 --> 01:44:35,960 ENERGY EXPENDITURE. 2294 01:44:35,960 --> 01:44:36,760 I.E. IT'S OBLIGATORY COMPONENT, 2295 01:44:36,760 --> 01:44:39,920 YOU HAVE TO SURVIVE, TO HAVE 60, 2296 01:44:39,920 --> 01:44:45,200 70% OF YOUR BASIC METABOLIC 2297 01:44:45,200 --> 01:44:45,960 RATE. 2298 01:44:45,960 --> 01:44:48,920 KNOW, THAT TURNS OUT TO BE 2299 01:44:48,920 --> 01:44:51,160 CLINICALLY MEANINGFUL RESEARCH 2300 01:44:51,160 --> 01:44:52,200 AND UNRESEARCHED SETTING, SO WE 2301 01:44:52,200 --> 01:44:54,280 DO THAT ALMOST EVERY DAY, 2302 01:44:54,280 --> 01:44:55,000 DIFFERENT WAYS TO MEASURE. 2303 01:44:55,000 --> 01:44:57,240 IT DOESN'T HAVE TO BE IN 2304 01:44:57,240 --> 01:44:58,520 METABOLIC CHAMBER, TURNS OUT. 2305 01:44:58,520 --> 01:45:01,400 WE DID THAT IN THE CLINIC, 2306 01:45:01,400 --> 01:45:02,920 WHETHER A PEDIATRIC UNIT OR 2307 01:45:02,920 --> 01:45:05,160 ADULT UNIT, USING THIS METABOLIC 2308 01:45:05,160 --> 01:45:05,360 CART. 2309 01:45:05,360 --> 01:45:10,040 THIS IS A CUTE PICTURE. 2310 01:45:10,040 --> 01:45:12,320 LITTLE BABY, WE MEASURED A 2311 01:45:12,320 --> 01:45:14,840 2-YEAR-OLD THREE WEEKS AGO. 2312 01:45:14,840 --> 01:45:16,760 THIS IS A VENTILATED HOOD OVER 2313 01:45:16,760 --> 01:45:19,240 AN INFANT, THEY CAN BE ASLEEP 2314 01:45:19,240 --> 01:45:19,800 HOPEFULLY, BECAUSE OTHERWISE 2315 01:45:19,800 --> 01:45:22,720 THEY WOULD BE IN CRYING IN THERE 2316 01:45:22,720 --> 01:45:27,000 AND NOT A METABOLIC RATE AT ALL, 2317 01:45:27,000 --> 01:45:28,200 OR ADULT UNIT, ESSENTIALLY THIS 2318 01:45:28,200 --> 01:45:30,400 IS THE INSTRUMENT THAT WE 2319 01:45:30,400 --> 01:45:31,120 MEASURE. 2320 01:45:31,120 --> 01:45:34,160 SAME PRINCIPLE EXCEPT WITH THE 2321 01:45:34,160 --> 01:45:36,320 SMALL DEAD SPACE, 7 LITERS OF 2322 01:45:36,320 --> 01:45:42,760 DEAD SPACE RATHER THAN 27,000 2323 01:45:42,760 --> 01:45:43,040 LITERS. 2324 01:45:43,040 --> 01:45:47,120 THE RATE IS DONE FASTED 2325 01:45:47,120 --> 01:45:52,040 OVERNIGHT, SUPINE, DIM LIGHT, 2326 01:45:52,040 --> 01:45:53,360 THERMONEUTRAL, WHAT I SPENT THE 2327 01:45:53,360 --> 01:45:55,440 LAST SIX YEARS TRYING TO FIGURE 2328 01:45:55,440 --> 01:46:00,400 THAT OUT, HAS TO BE QUIET, 2329 01:46:00,400 --> 01:46:01,280 MEASURING 25 TO 35 MINUTES BUT 2330 01:46:01,280 --> 01:46:07,320 IN A BABY I'M HAPPY WITH 10. 2331 01:46:07,320 --> 01:46:07,960 THERE ARE DIFFERENT 2332 01:46:07,960 --> 01:46:10,680 CONFIGURATION, WITH THE BABY WE 2333 01:46:10,680 --> 01:46:12,120 DIDN'T USE A VENTILATOR BUT CAN 2334 01:46:12,120 --> 01:46:13,200 WITH PATIENTS. 2335 01:46:13,200 --> 01:46:14,400 THE BIGGEST QUESTION THAT COMES 2336 01:46:14,400 --> 01:46:16,440 OUT, WE HAVE THE MEASUREMENT OF 2337 01:46:16,440 --> 01:46:18,600 BASIC METABOLIC RATE, IS IT 2338 01:46:18,600 --> 01:46:20,360 NORMAL, HYPER, OR HYPO. 2339 01:46:20,360 --> 01:46:22,080 CLINICALLY WE HAVE TO DETERMINE 2340 01:46:22,080 --> 01:46:24,400 THAT STATUS OF METABOLIC RATE. 2341 01:46:24,400 --> 01:46:27,440 TURNS OUT IT'S ONE OF THE LAST 2342 01:46:27,440 --> 01:46:32,400 LINEAR SYSTEMS LEFT THAT YOU 2343 01:46:32,400 --> 01:46:34,680 ASSOCIATE THESE 88 INDIVIDUALS, 2344 01:46:34,680 --> 01:46:35,960 RESTING ENERGY EXPENDITURE, 2345 01:46:35,960 --> 01:46:37,880 BASIC METABOLIC RATE, VERSUS 2346 01:46:37,880 --> 01:46:39,920 FAT-FREE MASS OR LEAN BODY MASS 2347 01:46:39,920 --> 01:46:43,120 MEASURED BY BODY COMPOSITION 2348 01:46:43,120 --> 01:46:45,600 METHODS, DEXA OR UNDERWATER, IT 2349 01:46:45,600 --> 01:46:49,320 HAS AMAZING LINEAR ASSOCIATION, 2350 01:46:49,320 --> 01:46:50,120 WHETHER YOU'RE LEAN, OBESE, 2351 01:46:50,120 --> 01:46:51,520 MALES AND FEMALES. 2352 01:46:51,520 --> 01:46:54,480 THEY ALL FOLLOW THIS LINEAR 2353 01:46:54,480 --> 01:46:55,000 LINE. 2354 01:46:55,000 --> 01:46:55,800 THERE'S OBVIOUSLY STANDARD 2355 01:46:55,800 --> 01:46:58,440 DEVIATIONS, WE CAN CALL THESE 2356 01:46:58,440 --> 01:46:59,760 THREE INDIVIDUALS HYPER 2357 01:46:59,760 --> 01:47:00,960 METABOLIC AND THEN MAYBE ONE OR 2358 01:47:00,960 --> 01:47:07,080 TWO INDIVIDUALS MAY BE HYPO 2359 01:47:07,080 --> 01:47:07,480 METABOLIC, THAT'S 2360 01:47:07,480 --> 01:47:08,120 CROSS-SECTIONALLY WE CAN 2361 01:47:08,120 --> 01:47:09,840 DETERMINE THAT, THIS IS MY 2362 01:47:09,840 --> 01:47:11,040 DATABASE AT VANDERBILT BEFORE I 2363 01:47:11,040 --> 01:47:12,720 CAME HERE. 2364 01:47:12,720 --> 01:47:14,800 WE HAD OVER 200 SUBJECTS. 2365 01:47:14,800 --> 01:47:17,920 SO THIS DEMONSTRATES TWO THINGS. 2366 01:47:17,920 --> 01:47:20,200 ONE IS THAT FAT-FREE MASS HAS 2367 01:47:20,200 --> 01:47:22,960 MUCH HIGHER CONTRIBUTION PER 2368 01:47:22,960 --> 01:47:25,920 KILOGRAMS VERSUS FAT MASS, ABOUT 2369 01:47:25,920 --> 01:47:28,000 4:1, REPRODUCED BY MANY LABS. 2370 01:47:28,000 --> 01:47:28,480 CROSS-SECTIONALLY. 2371 01:47:28,480 --> 01:47:33,040 THE OTHER THING THAT ALWAYS USES 2372 01:47:33,040 --> 01:47:35,080 AS EXAMPLE FOR MY PATIENTS, IF 2373 01:47:35,080 --> 01:47:37,640 I'M OVERWEIGHT I MUST HAVE A 2374 01:47:37,640 --> 01:47:39,280 LOWER METABOLIC RATE, AND THIS 2375 01:47:39,280 --> 01:47:40,800 SHOWS PER KILOGRAM OF FAT MASS 2376 01:47:40,800 --> 01:47:44,120 BY AND LARGE YOU HAVE HIGHER 2377 01:47:44,120 --> 01:47:46,600 METABOLIC RATE, FAT MASS 2378 01:47:46,600 --> 01:47:48,360 CONTRIBUTES TO METABOLIC RATE, 2379 01:47:48,360 --> 01:47:49,000 NOT NEGATIVELY, OKAY? 2380 01:47:49,000 --> 01:47:52,560 AND SO WE CAN ALSO APPLY TO 2381 01:47:52,560 --> 01:47:54,720 CLINICAL COMPUTATION, OLDER 2382 01:47:54,720 --> 01:48:01,080 STUDY WE DID THAT COMPARED 2383 01:48:01,080 --> 01:48:02,480 PATIENTS UNDER DIALYSIS, 2384 01:48:02,480 --> 01:48:05,440 PERITONEAL OR CHRONIC RENAL, 2385 01:48:05,440 --> 01:48:07,760 VERSUS CHRONIC RENAL FAILURE 2386 01:48:07,760 --> 01:48:09,600 PATIENTS, NON-DIALYSIS, CAN PLOT 2387 01:48:09,600 --> 01:48:13,320 RESTING METABOLIC RATE VERSUS 2388 01:48:13,320 --> 01:48:15,960 FAT-FREE MASS, CHRONIC RENAL 2389 01:48:15,960 --> 01:48:18,560 FAILURE HAS LOWER RESTING 2390 01:48:18,560 --> 01:48:20,920 METABOLIC RATE PER KILOGRAM 2391 01:48:20,920 --> 01:48:22,640 VERSUS PATIENTS RECEIVING 2392 01:48:22,640 --> 01:48:24,080 DIALYSIS, PRESUMABLY DUE TO 2393 01:48:24,080 --> 01:48:29,440 PROTEIN LEAK THAT THEY HAVE TO 2394 01:48:29,440 --> 01:48:32,040 REPLENISH, HIGHER PROTEIN 2395 01:48:32,040 --> 01:48:32,320 TURNOVERS. 2396 01:48:32,320 --> 01:48:35,960 THIS IS NON-DIALYSIS DAY, YOU 2397 01:48:35,960 --> 01:48:38,280 CAN PLOT THE GROUP-WISE 2398 01:48:38,280 --> 01:48:39,120 DIFFERENCE. 2399 01:48:39,120 --> 01:48:42,640 FIRST STUDY WITH CLINICAL TRIAL, 2400 01:48:42,640 --> 01:48:44,080 RECEPTOR AGONIST, APPROVED BUT 2401 01:48:44,080 --> 01:48:46,280 NOT FOR OBESITY RESEARCH, OR FOR 2402 01:48:46,280 --> 01:48:49,240 OBESITY TREATMENT BUT FOR 2403 01:48:49,240 --> 01:48:51,280 ANOTHER INDICATION, BUT THIS IS 2404 01:48:51,280 --> 01:48:54,120 WHEN WE TOOK A COMPOUND FROM 2405 01:48:54,120 --> 01:48:56,440 RHYTHM PHARMACEUTICAL AND WE 2406 01:48:56,440 --> 01:48:59,200 KNOW THE MECHANISM MAY AFFECT 2407 01:48:59,200 --> 01:49:02,000 METABOLISM AND DID STUDY IN 2408 01:49:02,000 --> 01:49:04,040 METABOLIC CHAMBER, 48 HOUR 2409 01:49:04,040 --> 01:49:08,360 RANDOM CROSSOVER STUDY IN I 2410 01:49:08,360 --> 01:49:10,200 THINK 12 INDIVIDUALS, MEASURED 2411 01:49:10,200 --> 01:49:11,640 RESTORATIONING METABOLIC RATE IN 2412 01:49:11,640 --> 01:49:15,680 THE DAY, COMPARED DRUG VERSUS 2413 01:49:15,680 --> 01:49:17,440 PLACEBO, AND CROSSOVER FASHION 2414 01:49:17,440 --> 01:49:22,120 WERE ABLE TO SHOW THE CHAMBER 2415 01:49:22,120 --> 01:49:23,560 MEASUREMENT, RESTING METABOLIC 2416 01:49:23,560 --> 01:49:25,120 RATE WAS SLIGHTLY HIGHER, ABOUT 2417 01:49:25,120 --> 01:49:28,920 6% HIGHER WITH THE DRUG VERSUS 2418 01:49:28,920 --> 01:49:30,680 PLACEBO, THIS IS, AGAIN, WITH 2419 01:49:30,680 --> 01:49:34,960 INDIVIDUAL DIFFERENCE OF 6% WITH 2420 01:49:34,960 --> 01:49:38,360 12 INDIVIDUALS, SHOWS YOU THE 2421 01:49:38,360 --> 01:49:39,640 PRECISION OF THE MEASUREMENT AND 2422 01:49:39,640 --> 01:49:42,400 LEVERAGE, BE ABLE TO STUDY 2423 01:49:42,400 --> 01:49:44,040 PHYSIOLOGY, WHOLE HUMANS. 2424 01:49:44,040 --> 01:49:45,600 AND THIS IS THE SPAGHETTI PLOT. 2425 01:49:45,600 --> 01:49:48,640 AND WE ALSO DID A HOOD 2426 01:49:48,640 --> 01:49:49,400 MEASUREMENT. 2427 01:49:49,400 --> 01:49:49,720 RIGHT HERE. 2428 01:49:49,720 --> 01:49:52,880 YOU CAN SEE BY GROUP DIFFERENCE 2429 01:49:52,880 --> 01:49:54,680 NOT MUCH, BUT THE PRECISION OF 2430 01:49:54,680 --> 01:49:55,520 THE CHAMBER MEASUREMENT IS 2431 01:49:55,520 --> 01:49:58,280 HIGHER, THEREFORE WE HAVE A 2432 01:49:58,280 --> 01:50:00,360 BETTER P-VALUE. 2433 01:50:00,360 --> 01:50:02,440 ANOTHER THING THAT GETS A LOT OF 2434 01:50:02,440 --> 01:50:06,480 INTEREST IS HOW THE BASIC 2435 01:50:06,480 --> 01:50:09,880 METABOLIC RATE CHANGES, IT'S NOT 2436 01:50:09,880 --> 01:50:10,120 STATIC. 2437 01:50:10,120 --> 01:50:11,400 BODY COMPOSITION CHANGES AS 2438 01:50:11,400 --> 01:50:15,000 WELL. 2439 01:50:15,000 --> 01:50:17,880 RESIDENT ENERGY EXPENDITURE 2440 01:50:17,880 --> 01:50:19,400 CHANGES WITH WEIGHT LOSS. 2441 01:50:19,400 --> 01:50:23,440 BEE DID THIS STUDY BY COMPARING 2442 01:50:23,440 --> 01:50:24,880 GASTRIC BYPASS SURGERY, 2443 01:50:24,880 --> 01:50:29,240 TREATMENT FOR SEVERE OBESITY, 2444 01:50:29,240 --> 01:50:32,400 VERSUS BIGGEST LOSER SORT OF 2445 01:50:32,400 --> 01:50:33,720 CONTESTANTS, WELL DOCUMENTED IN 2446 01:50:33,720 --> 01:50:36,320 THE SORT OF POPULAR SCIENCE 2447 01:50:36,320 --> 01:50:36,760 LITERATURE. 2448 01:50:36,760 --> 01:50:44,000 BUT WHEN WE DO THAT, YOU MAY NOT 2449 01:50:44,000 --> 01:50:45,360 SEE. 2450 01:50:45,360 --> 01:50:47,400 BIGGEST LOSER, 13 SUBJECTS, THEY 2451 01:50:47,400 --> 01:50:49,960 FOLLOW UP 7 MONTHS, MATCHED WITH 2452 01:50:49,960 --> 01:50:52,560 THE BARIATRIC SURGERY PATIENTS 2453 01:50:52,560 --> 01:50:54,240 WE FOLLOWED AT VANDERBILT, 13, 2454 01:50:54,240 --> 01:50:55,840 MATCHED FOR AGE AND BODY WEIGHT. 2455 01:50:55,840 --> 01:50:59,480 THEY CAME IN AT 140 KILOGRAMS, 2456 01:50:59,480 --> 01:51:05,920 135 KILOGRAMS, WELL MATCHED. 2457 01:51:05,920 --> 01:51:16,440 AND PERCENT BODY FAT WAS 50/50, 2458 01:51:20,920 --> 01:51:24,040 SEVEN MONTHS THE BIGGEST LOSER 2459 01:51:24,040 --> 01:51:27,200 LOST 35% OF THE BODY WEIGHT. 2460 01:51:27,200 --> 01:51:29,680 FAIRLY WELL MATCHED BUT WHEN YOU 2461 01:51:29,680 --> 01:51:30,480 MEASURE ENERGY EXPENDITURE, 2462 01:51:30,480 --> 01:51:34,080 VERSUS CHANGES IN BODY 2463 01:51:34,080 --> 01:51:37,720 COMPOSITION, THERE'S AN 2464 01:51:37,720 --> 01:51:40,520 ADAPTATION OF 400 CALORIES PER 2465 01:51:40,520 --> 01:51:43,000 DAY, THEY DROP BELOW PREDICTED 2466 01:51:43,000 --> 01:51:44,040 ENERGY EXPENDITURE ACCORDING TO 2467 01:51:44,040 --> 01:51:48,920 NO BODY COMPOSITION COMPARED TO 2468 01:51:48,920 --> 01:51:53,120 HALF OF THAT IN THE GASTRIC 2469 01:51:53,120 --> 01:51:55,760 BYPASS SURGERY. 2470 01:51:55,760 --> 01:51:57,000 TWELVE MONTHS LATER THE GASTRIC 2471 01:51:57,000 --> 01:51:59,600 SURGERY RETURNED TO ORIGINAL 2472 01:51:59,600 --> 01:52:03,280 LEVEL IN RESTING METABOLIC RATE, 2473 01:52:03,280 --> 01:52:04,400 METABOLIC ADAPTATION REDUCED TO 2474 01:52:04,400 --> 01:52:04,920 ALMOST NOTHING. 2475 01:52:04,920 --> 01:52:06,360 WE DIDN'T MEASURE IN BIGGEST 2476 01:52:06,360 --> 01:52:07,560 LOSERS BECAUSE THEY HAVE GONE 2477 01:52:07,560 --> 01:52:10,280 HOME AND WE CAN'T FOLLOW THEM 2478 01:52:10,280 --> 01:52:12,200 UP. 2479 01:52:12,200 --> 01:52:14,880 SO, THIS SHOWS WHAT I SHOWED 2480 01:52:14,880 --> 01:52:15,560 BEFORE. 2481 01:52:15,560 --> 01:52:18,080 IT TURNS OUT THAT THE 2482 01:52:18,080 --> 01:52:20,360 INDIVIDUAL, THIS IS INDIVIDUAL 2483 01:52:20,360 --> 01:52:22,560 METABOLIC ADAPTATION VERSUS THE 2484 01:52:22,560 --> 01:52:25,200 CHANGES IN THE ENERGY IMBALANCE 2485 01:52:25,200 --> 01:52:28,920 OR CALCULATED ENERGY INTAKE 2486 01:52:28,920 --> 01:52:30,200 VERSUS BODY COMPOSITION, AND 2487 01:52:30,200 --> 01:52:33,000 LEPTIN LEVEL, WHICH IS FAT 2488 01:52:33,000 --> 01:52:35,080 SECRETED, WHITE FAT SECRETED 2489 01:52:35,080 --> 01:52:36,800 HORMONE, SENSED BY THE BRAIN. 2490 01:52:36,800 --> 01:52:38,760 SO INDIVIDUAL LEVELS OF 2491 01:52:38,760 --> 01:52:40,840 METABOLIC ADAPTATION IS 2492 01:52:40,840 --> 01:52:44,760 CORRELATED TO BOTH MEASURES 2493 01:52:44,760 --> 01:52:48,520 SUGGESTING IT IS A PHYSIOLOGIC 2494 01:52:48,520 --> 01:52:50,120 MEASURE WE CAN MEASURE AND 2495 01:52:50,120 --> 01:52:50,360 PREDICT. 2496 01:52:50,360 --> 01:52:52,360 I WON'T TELL YOU THE OTHER 2497 01:52:52,360 --> 01:52:54,720 COMPONENTS, ALSO VERY IMPORTANT 2498 01:52:54,720 --> 01:52:56,320 BUT SMALLER IN SCALE, PHYSICAL 2499 01:52:56,320 --> 01:52:57,800 ACTIVITY, USING A LOT OF 2500 01:52:57,800 --> 01:53:01,480 WEARABLES TO DO THAT, AND 2501 01:53:01,480 --> 01:53:02,720 ADAPTIVE THERMOGENESIS WHICH 2502 01:53:02,720 --> 01:53:05,880 INCLUDE DIET INDUCED AND AS WELL 2503 01:53:05,880 --> 01:53:07,640 AS COLD INDUCED, WHEN WE TALK 2504 01:53:07,640 --> 01:53:09,160 ABOUT COLD WE MEASURE BROWN FAT, 2505 01:53:09,160 --> 01:53:11,280 SO I'M NOT GOING TO TELL YOU ANY 2506 01:53:11,280 --> 01:53:14,400 OF THAT BECAUSE THAT'S GOING TO 2507 01:53:14,400 --> 01:53:16,480 TAKE A LOT OF TIME. 2508 01:53:16,480 --> 01:53:19,320 HOPEFULLY THESE ARE MY NOTES, 2509 01:53:19,320 --> 01:53:21,960 AND CHEAT SHEET, TAKE HOME, THE 2510 01:53:21,960 --> 01:53:24,160 RESIDENT ENERGY EXPENDITURE IS A 2511 01:53:24,160 --> 01:53:26,120 MAJOR COMPONENT OF TOTAL ENERGY 2512 01:53:26,120 --> 01:53:28,160 EXPENDITURE, HIGHLY CORRELATED 2513 01:53:28,160 --> 01:53:31,800 WITH BODY COMPOSITION, FAT FREE 2514 01:53:31,800 --> 01:53:33,240 MASS, NOT STATIC, IT'S ACUTE AND 2515 01:53:33,240 --> 01:53:36,520 RAPID CHANGES IN BODY WEIGHT CAN 2516 01:53:36,520 --> 01:53:41,240 RESULT IN METABOLIC ADAPTATION, 2517 01:53:41,240 --> 01:53:42,000 PHARMACEUTICALS OR 2518 01:53:42,000 --> 01:53:44,080 NUTRICEUTICALS CAN ALTER BASIC 2519 01:53:44,080 --> 01:53:47,720 METABOLIC RATE IN ACUTE AND 2520 01:53:47,720 --> 01:53:49,760 CHRONIC CONDITIONS, THIS IS A 2521 01:53:49,760 --> 01:53:52,720 TEXTBOOK FROM 1970s SHOWING 2522 01:53:52,720 --> 01:53:54,920 METABOLIC RATE DO ADJUST TO 2523 01:53:54,920 --> 01:53:55,680 CLINICAL CONDITIONS BOTH IN 2524 01:53:55,680 --> 01:53:58,840 TERMS OF FEVER AS WELL AS 2525 01:53:58,840 --> 01:54:00,160 PARTIAL STARVATION, WHICH IS 2526 01:54:00,160 --> 01:54:02,120 REALLY NOT MUCH DIFFERENT THAN 2527 01:54:02,120 --> 01:54:04,520 THE BIGGEST LOSER CASE, RIGHT? 2528 01:54:04,520 --> 01:54:07,160 THEY DO DECREASE METABOLIC RATE 2529 01:54:07,160 --> 01:54:08,600 WITH PARTIAL STARVATION, 2530 01:54:08,600 --> 01:54:12,400 INCREASES WITH BURNS OR FEVER. 2531 01:54:12,400 --> 01:54:15,360 SO WE'RE VERY INTERESTED IN 2532 01:54:15,360 --> 01:54:17,000 DISEASE CONDITIONS, CANCER, HIV, 2533 01:54:17,000 --> 01:54:20,680 COVID-19, WHETHER WEIGHT LOSS 2534 01:54:20,680 --> 01:54:22,560 CONDITIONS OR AGE-SPECIFIC, YOU 2535 01:54:22,560 --> 01:54:24,320 KNOW, INTERVENTIONS IN THAT, AND 2536 01:54:24,320 --> 01:54:27,600 AS WELL AS DIFFERENT MEDICATIONS 2537 01:54:27,600 --> 01:54:30,640 USED FOR DIABETES, OBESITY, 2538 01:54:30,640 --> 01:54:31,520 SSRIs, VACCINES. 2539 01:54:31,520 --> 01:54:33,520 SO OUR PRINCIPLE IS TO MEASURE 2540 01:54:33,520 --> 01:54:34,720 EVERY COMPONENT POSSIBLE AND 2541 01:54:34,720 --> 01:54:36,000 WHAT I'VE SHOWN IS ONLY ONE 2542 01:54:36,000 --> 01:54:38,120 ASPECT OF WHAT WE DO. 2543 01:54:38,120 --> 01:54:41,040 WE MEASURE BODY COMPOSITION, WE 2544 01:54:41,040 --> 01:54:44,040 ALSO MEASURE PET-CT CAN BROWN 2545 01:54:44,040 --> 01:54:46,840 FAT AND USE ACCELEROMETER AND 2546 01:54:46,840 --> 01:54:47,960 WEARABLES TO MEASURE PHYSICAL 2547 01:54:47,960 --> 01:54:53,080 ACTIVITY AND STILL HAVE TROUBLE 2548 01:54:53,080 --> 01:54:53,880 MEASURING PHYSICAL ACTIVITY. 2549 01:54:53,880 --> 01:55:00,000 I WANT TO ACKNOWLEDGE THE PEOPLE 2550 01:55:00,000 --> 01:55:02,400 IN MY LAB, CURRENT LAB, AND 2551 01:55:02,400 --> 01:55:03,920 PREVIOUS MEMBERS HERE. 2552 01:55:03,920 --> 01:55:06,080 WITH THAT, I'D BE HAPPY TO 2553 01:55:06,080 --> 01:55:06,560 ANSWER ANY QUESTIONS. 2554 01:55:06,560 --> 01:55:08,960 [APPLAUSE] 2555 01:55:08,960 --> 01:55:15,000 2556 01:55:15,000 --> 01:55:17,840 2557 01:55:17,840 --> 01:55:23,640 >> DO YOU USE OPTICAL IMAGING 2558 01:55:23,640 --> 01:55:26,920 DEVICES -- (INAUDIBLE). 2559 01:55:26,920 --> 01:55:29,240 >> SO LET ME REPEAT THE 2560 01:55:29,240 --> 01:55:30,520 QUESTION FOR PEOPLE ONLINE. 2561 01:55:30,520 --> 01:55:32,520 YOU DIDN'T USE THE MICROPHONE. 2562 01:55:32,520 --> 01:55:33,600 YOU'RE THE HOST, YOU CAN BREAK 2563 01:55:33,600 --> 01:55:34,000 THE RULES. 2564 01:55:34,000 --> 01:55:38,880 THE QUESTION IS DO I USE ANY 2565 01:55:38,880 --> 01:55:39,760 OPTICAL MEASUREMENTS 2566 01:55:39,760 --> 01:55:42,560 SIMULTANEOUSLY IN THE CHAMBER. 2567 01:55:42,560 --> 01:55:45,400 WE DO HAVE OPTICAL CAMERA, AND 2568 01:55:45,400 --> 01:55:46,640 ACTUALLY WITH SLIGHT INFRARED TO 2569 01:55:46,640 --> 01:55:48,040 IT, MOSTLY FOR SAFETY. 2570 01:55:48,040 --> 01:55:52,320 WE DON'T RECORD IT FOR SUBJECT 2571 01:55:52,320 --> 01:55:53,640 PRIVACY REASONS. 2572 01:55:53,640 --> 01:55:57,560 BUT THEY ARE METABOLIC CHAMBERS 2573 01:55:57,560 --> 01:55:59,000 WITH OPTICAL IMAGING FOR, YOU 2574 01:55:59,000 --> 01:56:00,960 KNOW, RECORD THE TYPE OF 2575 01:56:00,960 --> 01:56:02,800 ACTIVITIES AND INTENSITY OF 2576 01:56:02,800 --> 01:56:03,080 ACTIVITIES. 2577 01:56:03,080 --> 01:56:05,320 AND LAST BUT NOT THE LEAST, WE 2578 01:56:05,320 --> 01:56:08,320 DO HAVE WHEN WE DO BROWN FAT 2579 01:56:08,320 --> 01:56:11,680 STUDIES, WE DO AN IR IMAGING OF 2580 01:56:11,680 --> 01:56:13,320 SURFACE BODY TEMPERATURE, BUT 2581 01:56:13,320 --> 01:56:15,720 STATIC IMAGE. 2582 01:56:15,720 --> 01:56:18,800 2583 01:56:18,800 --> 01:56:19,440 2584 01:56:19,440 --> 01:56:21,120 >> I'M CURIOUS ABOUT THE RADAR 2585 01:56:21,120 --> 01:56:22,280 MEASUREMENT OF ACTIVITY AND HOW 2586 01:56:22,280 --> 01:56:24,320 IT'S ONLY A SINGLE PARAMETER 2587 01:56:24,320 --> 01:56:28,120 BECAUSE I WONDER IF YOU LOOKED 2588 01:56:28,120 --> 01:56:29,720 AT THE DIFFERENCES IN ENERGY 2589 01:56:29,720 --> 01:56:32,200 EXPENDITURE AS A FUNCTION OF -- 2590 01:56:32,200 --> 01:56:33,560 >>YES, YES. 2591 01:56:33,560 --> 01:56:35,520 ANOTHER VERY GOOD QUESTION. 2592 01:56:35,520 --> 01:56:37,160 SO, THE RADAR SENSOR HAS REALLY 2593 01:56:37,160 --> 01:56:38,600 BEEN IN THERE FOR HISTORICAL 2594 01:56:38,600 --> 01:56:41,760 REASONS BECAUSE A LOT OF THE 2595 01:56:41,760 --> 01:56:43,840 TECHNIQUE TRYING TO SEPARATE THE 2596 01:56:43,840 --> 01:56:45,720 DIFFERENT COMPONENTS IS RADAR 2597 01:56:45,720 --> 01:56:49,120 BASED, BUT WE HAVE A LOT MORE 2598 01:56:49,120 --> 01:56:54,360 WEARABLE SENSORS TO GET MORE 2599 01:56:54,360 --> 01:56:57,960 QUANTITATIVE MEASUREMENTS, IT'S 2600 01:56:57,960 --> 01:56:58,480 A QUANTITATIVE/QUALITATIVE 2601 01:56:58,480 --> 01:57:01,120 MEASUREMENT, RIGHT? 2602 01:57:01,120 --> 01:57:02,880 BUT ACTUALLY BEFORE I CAME HERE 2603 01:57:02,880 --> 01:57:07,120 IN 2006 I HAD TWO R01s AT 2604 01:57:07,120 --> 01:57:08,120 VANDERBILT, DEVELOPING WEARABLE 2605 01:57:08,120 --> 01:57:09,040 SENSORS. 2606 01:57:09,040 --> 01:57:13,080 SO IF YOU WONDER HOW YOUR APPLE 2607 01:57:13,080 --> 01:57:13,720 WATCH ESTIMATES YOUR CALORIES, 2608 01:57:13,720 --> 01:57:20,080 WE HAD A PART IN THAT. 2609 01:57:20,080 --> 01:57:22,480 >> IN TERMS OF BODY 2610 01:57:22,480 --> 01:57:23,680 COMPOSITION, HOW MUCH EFFECT 2611 01:57:23,680 --> 01:57:26,520 DOES MUSCLE MASS HAVE ON EITHER 2612 01:57:26,520 --> 01:57:27,600 ENERGY EXPENDITURE OR BMR? 2613 01:57:27,600 --> 01:57:31,320 >>VERY GOOD QUESTION. 2614 01:57:31,320 --> 01:57:33,840 SO, IT TURNS OUT MUSCLE MASS AT 2615 01:57:33,840 --> 01:57:37,120 RESTING LEVEL CONTRIBUTES TO 2616 01:57:37,120 --> 01:57:39,520 VERY LITTLE OF YOUR BASIC 2617 01:57:39,520 --> 01:57:41,840 METABOLIC RATE BUT ORGAN SIZE IS 2618 01:57:41,840 --> 01:57:44,080 HIGHLY CORRELATED WITH MUSCLE 2619 01:57:44,080 --> 01:57:44,280 MASS. 2620 01:57:44,280 --> 01:57:47,440 IF WE COULD MEASURE ORGAN SIZE 2621 01:57:47,440 --> 01:57:48,720 AND ORGAN-SPECIFIC METABOLIC 2622 01:57:48,720 --> 01:57:53,760 RATE, THAT'S THE HOLY GRAIL OF 2623 01:57:53,760 --> 01:57:55,640 MEASURING THE METABOLIC 2624 01:57:55,640 --> 01:57:58,160 DIFFERENCES, THAT HAS MUCH 2625 01:57:58,160 --> 01:58:00,040 HIGHER CONTRIBUTION, FOR EXAMPLE 2626 01:58:00,040 --> 01:58:04,400 BRAIN, LIVER, HEART, AND KIDNEY 2627 01:58:04,400 --> 01:58:06,440 CONTRIBUTES TO ABOUT 60% OF 2628 01:58:06,440 --> 01:58:07,880 METABOLIC RATE, BUT ON THE 2629 01:58:07,880 --> 01:58:11,320 INDIVIDUAL LEVEL IT'S ACTUALLY 2630 01:58:11,320 --> 01:58:12,280 VERY DIFFICULT TO DETERMINE. 2631 01:58:12,280 --> 01:58:22,360 P. . 2632 01:58:23,120 --> 01:58:23,760 >> OZEMPIC, METABOLIC RATE? 2633 01:58:23,760 --> 01:58:24,880 >> ANOTHER INTERESTING 2634 01:58:24,880 --> 01:58:25,160 QUESTION. 2635 01:58:25,160 --> 01:58:30,560 WE'RE DOING A FEW OF THOSE 2636 01:58:30,560 --> 01:58:32,320 STUDIES, BUT IT TURNS OUT MOSTLY 2637 01:58:32,320 --> 01:58:34,960 REGULATING ON THE ENERGY INTAKE 2638 01:58:34,960 --> 01:58:36,360 SIDE, NOT SO MUCH ENERGY 2639 01:58:36,360 --> 01:58:38,640 EXPENDITURE BUT HOW DOES THAT 2640 01:58:38,640 --> 01:58:44,240 IMPACT METABOLIC ADAPTATIONS? 2641 01:58:44,240 --> 01:58:46,320 WE DON'T KNOW. 2642 01:58:46,320 --> 01:58:46,760 OKAY, THANK YOU. 2643 01:58:46,760 --> 01:58:59,400 [APPLAUSE] 2644 01:58:59,400 --> 01:59:01,280 INVENTION, DESIGN, DEVELOPMENT 2645 01:59:01,280 --> 01:59:03,680 AND APPLICATION OF BIOMEDICAL 2646 01:59:03,680 --> 01:59:06,080 OPTICS AND INSTRUMENTATION TO 2647 01:59:06,080 --> 01:59:15,560 BROAD RANGE OF INTERDISCIPLINARY 2648 01:59:15,560 --> 01:59:15,960 RESEARCH PROGRAMS. 2649 01:59:15,960 --> 01:59:17,240 >> THANKS YOU FOR THE 2650 01:59:17,240 --> 01:59:18,200 INTRODUCTION AND INVITING ME TO 2651 01:59:18,200 --> 01:59:22,160 PRESENT THE WORK IN DEVELOPING 2652 01:59:22,160 --> 01:59:26,880 ADAPTIVE OPTIC TECHNIQUES TO 2653 01:59:26,880 --> 01:59:34,680 EXPLORE RETINAL FUNCTION AND 2654 01:59:34,680 --> 01:59:37,840 DYSFUNCTION. 2655 01:59:37,840 --> 01:59:43,400 I'D LIKE TO START BY RECOGNIZING 2656 01:59:43,400 --> 01:59:45,600 COLLABORATORS, ESPECIALLY HERE 2657 01:59:45,600 --> 01:59:48,920 AT NATIONAL EYE INSTITUTE, 2658 01:59:48,920 --> 01:59:52,200 INCLUDING KATHY KUKROS AND WORK 2659 01:59:52,200 --> 01:59:53,040 WORKING WITH RETINAL DISEASES 2660 01:59:53,040 --> 01:59:56,240 AND JOHNNY TAM WHO WE'RE HELPING 2661 01:59:56,240 --> 01:59:58,000 BUILD SOME A.O. SYSTEMS. 2662 01:59:58,000 --> 02:00:01,240 HE'S DONE SOME NICE WORK USING 2663 02:00:01,240 --> 02:00:04,760 ICG TECHNIQUE TO LOOK AT 2664 02:00:04,760 --> 02:00:06,080 METABOLISM AND RPE CELLS. 2665 02:00:06,080 --> 02:00:10,160 SO, I WORK FOR THE OFFICE OF 2666 02:00:10,160 --> 02:00:11,680 SCIENCE AND ENGINEERING LABS, 2667 02:00:11,680 --> 02:00:16,160 AND OUR PRIMARY OUTPUT IS 2668 02:00:16,160 --> 02:00:17,120 REGULATORY SCIENCE TOOLS, 2669 02:00:17,120 --> 02:00:18,720 REGULATORY SCIENCE IS DEFINED BY 2670 02:00:18,720 --> 02:00:21,760 FDA AS DEVELOPMENT OF NEW TOOLS, 2671 02:00:21,760 --> 02:00:25,480 STANDARDS, APPROACHES TO ASSESS 2672 02:00:25,480 --> 02:00:27,520 QUALITY AND PERFORMANCE OF 2673 02:00:27,520 --> 02:00:28,560 FDA-REGULATED PROCESS. 2674 02:00:28,560 --> 02:00:30,720 IN THE RESEARCH ARM, OUR OFFICE, 2675 02:00:30,720 --> 02:00:33,400 WE HAVE ESTABLISHED A CATALOG OF 2676 02:00:33,400 --> 02:00:39,640 REGULATORY SCIENCE TOOLS THAT 2677 02:00:39,640 --> 02:00:44,440 INCLUDE PHANTOMS, MODELS, USEFUL 2678 02:00:44,440 --> 02:00:45,520 IN THE INDUSTRY. 2679 02:00:45,520 --> 02:00:47,080 IF YOU CERTAIN REGULATORY 2680 02:00:47,080 --> 02:00:52,640 SCIENCE TOOLS CATALOG FROM THE 2681 02:00:52,640 --> 02:00:53,520 FDA.GOV WEBSITE YOU'LL ACCESS 2682 02:00:53,520 --> 02:00:54,720 THIS AND LOOK AT TOOLS WE'VE 2683 02:00:54,720 --> 02:00:56,800 DEVELOPED IN THE OFFICE. 2684 02:00:56,800 --> 02:00:58,200 WE BELIEVE THESE TOOLS WILL 2685 02:00:58,200 --> 02:01:01,280 EXPAND THE SCOPE OF INNOVATEIVE 2686 02:01:01,280 --> 02:01:03,160 SCIENCE-BASED APPROACHES TO 2687 02:01:03,160 --> 02:01:07,240 IMPROVE DEVELOPMENT OF EMERGING 2688 02:01:07,240 --> 02:01:09,160 MEDICAL TECHNOLOGIES. 2689 02:01:09,160 --> 02:01:14,880 WE'VE ESTABLISHED ADAPTIVE 2690 02:01:14,880 --> 02:01:17,560 OPTICS PROGRAM AT FDA, TO ALLOW 2691 02:01:17,560 --> 02:01:20,120 CELLULAR RESOLUTION OF RETINA BY 2692 02:01:20,120 --> 02:01:21,400 CORRECTING FOR OCULAR 2693 02:01:21,400 --> 02:01:25,360 ABERRATIONS THAT OCCUR IN ALL OF 2694 02:01:25,360 --> 02:01:27,760 US. 2695 02:01:27,760 --> 02:01:30,040 THE PROGRAM GOALS, ONE TO 2696 02:01:30,040 --> 02:01:32,120 DEVELOP PHANTOM-BASED EYE MODELS 2697 02:01:32,120 --> 02:01:34,760 AND NON-CLINICAL TOOLS FOR 2698 02:01:34,760 --> 02:01:36,480 PERFORMANCE ASSESSMENT, AND 2699 02:01:36,480 --> 02:01:39,120 THESE RETINAL IMAGING DEVICES 2700 02:01:39,120 --> 02:01:47,360 THAT USE ADAPTIVE OPTICS INCLUDE 2701 02:01:47,360 --> 02:01:48,160 DEFORMABLE MIRROR, PERSONALIZED, 2702 02:01:48,160 --> 02:01:49,560 EVERY PATIENT SUBJECT IN FRONT 2703 02:01:49,560 --> 02:01:52,080 OF THEM THEY ADAPT TOP. 2704 02:01:52,080 --> 02:01:54,800 AND THAT CAUSES SOME CHALLENGE 2705 02:01:54,800 --> 02:01:55,920 FOR PERFORMANCE ASSESSMENT. 2706 02:01:55,920 --> 02:01:59,600 THE SECOND PART OF OUR PROGRAM 2707 02:01:59,600 --> 02:02:01,920 IS TO INVENT, DEVELOP, VALIDATE 2708 02:02:01,920 --> 02:02:04,880 A.O. BASED STRUCTURAL AND 2709 02:02:04,880 --> 02:02:06,400 FUNCTIONAL BIOMARKERS FOR EARLY 2710 02:02:06,400 --> 02:02:14,000 DISEASE DIAGNOSIS FOR USE AS 2711 02:02:14,000 --> 02:02:14,640 PRIMARY CLINICAL ENDPOINTS. 2712 02:02:14,640 --> 02:02:18,360 I'D LIKE TO TAKE YOU ON A TOUR 2713 02:02:18,360 --> 02:02:22,720 OF THE RETINA AND NOTE A.O. 2714 02:02:22,720 --> 02:02:23,720 TECHNIQUES WE'RE USING TO 2715 02:02:23,720 --> 02:02:25,800 EXPLORE THE RETINA. 2716 02:02:25,800 --> 02:02:27,200 THIS IS A CROSS-SECTION TAKEN 2717 02:02:27,200 --> 02:02:28,520 FROM AN A.O. SYSTEM. 2718 02:02:28,520 --> 02:02:32,040 YOU CAN SEE THE LAYERS THERE, 2719 02:02:32,040 --> 02:02:34,560 THE MAIN CELLULAR COMPONENTS OF 2720 02:02:34,560 --> 02:02:38,600 RETINA INCLUDE GANGLION CELLS, 2721 02:02:38,600 --> 02:02:40,240 IMPORTANT IN GLAUCOMA, BIPOLAR 2722 02:02:40,240 --> 02:02:41,720 CELLS AND LIGHT SENSITIVE 2723 02:02:41,720 --> 02:02:44,720 PHOTORECEPTORS AND RPE, WHICH IS 2724 02:02:44,720 --> 02:02:48,440 A MONOLAYER, AND THROUGH WHICH 2725 02:02:48,440 --> 02:02:52,680 NUTRIENTS FROM THE CHORIOCAP 2726 02:02:52,680 --> 02:02:56,920 LARULARIS PASS TO PHOTO 2727 02:02:56,920 --> 02:02:57,480 PHOTORECEPTORS. 2728 02:02:57,480 --> 02:03:03,360 WE'RE GOING TO EXPLORE 2729 02:03:03,360 --> 02:03:05,240 MACROPHAGE CELLS AT THE TOP OF 2730 02:03:05,240 --> 02:03:10,880 THE RETINA, FURTHER IN THE 2731 02:03:10,880 --> 02:03:17,880 RETINA, WE'LL LOOK AT GLAUCOMA. 2732 02:03:17,880 --> 02:03:21,960 A NEW TECHNIQUE, YOU CAN MEASURE 2733 02:03:21,960 --> 02:03:23,240 FUNCTION OF INDIVIDUAL 2734 02:03:23,240 --> 02:03:24,800 PHOTORECEPTORS, RPE CELL, I 2735 02:03:24,800 --> 02:03:27,960 NOTED WE'LL LOOK AT ORGANELLE 2736 02:03:27,960 --> 02:03:37,600 MOTILITY AND LATEST WORK IN 2737 02:03:37,600 --> 02:03:38,560 IMAGING CHORIOCAPILLARIS. 2738 02:03:38,560 --> 02:03:40,640 MACROPHAGES MAKE UP 15% OF BRAIN 2739 02:03:40,640 --> 02:03:47,920 CELLS, PRIMARY FUNCTION IS IN 2740 02:03:47,920 --> 02:03:56,880 IMMUNE RESPONSE TO INJURY. 2741 02:03:56,880 --> 02:04:02,760 MY UNDERSTANDING, WONG DID SOME 2742 02:04:02,760 --> 02:04:05,120 NICE WORK AT NIH, AND NEI, IN 2743 02:04:05,120 --> 02:04:06,840 STAINING THESE CELLS IN THE 2744 02:04:06,840 --> 02:04:08,360 RETINA WHICH OCCUR PRIMARILY IN 2745 02:04:08,360 --> 02:04:16,200 THE PLEXIFORM LAYERS AS YOU CAN 2746 02:04:16,200 --> 02:04:21,400 SEE HERE IN THIS FLUORESCENT 2747 02:04:21,400 --> 02:04:27,400 IMAGE, A PORTION OCCUR AT THE 2748 02:04:27,400 --> 02:04:27,640 MEMBRANE. 2749 02:04:27,640 --> 02:04:29,600 UNTIL RECENTLY CELLS HAVE BEEN 2750 02:04:29,600 --> 02:04:31,680 ONLY VISUALIZED WITH FLUORESCENT 2751 02:04:31,680 --> 02:04:34,160 MARKERS, EXOGENOUS DYES OR 2752 02:04:34,160 --> 02:04:35,640 TRANSGENIC LABELING. 2753 02:04:35,640 --> 02:04:37,560 ANOTHER RECENT STUDY FOUND THAT 2754 02:04:37,560 --> 02:04:39,120 IN THE RESTING STATE WITHIN 10 2755 02:04:39,120 --> 02:04:45,760 MICRONS OF THE RETINAL SURFACE, 2756 02:04:45,760 --> 02:04:51,120 82% ARE MICROGLIA, COMPARED TO 2757 02:04:51,120 --> 02:04:53,680 69% VITREOUS MACROPHAGE CELLS, 2758 02:04:53,680 --> 02:04:55,280 9% PARA VASCULAR MACROPHAGES. 2759 02:04:55,280 --> 02:04:58,680 YOU'LL SEE THEM CALLED 2760 02:04:58,680 --> 02:05:03,640 MACROPHAGE-LIKE CELLS, BUT THEY 2761 02:05:03,640 --> 02:05:04,720 ARE PREDOMINANTLY MICROGLIA. 2762 02:05:04,720 --> 02:05:06,640 WE UNDERTOOK A STUDY USING 2763 02:05:06,640 --> 02:05:07,680 ADAPTIVE OPTICS TO RESOLVE CELLS 2764 02:05:07,680 --> 02:05:09,880 IN THE RETINA AND WE WERE ABLE 2765 02:05:09,880 --> 02:05:13,160 TO DO THAT AND MAP THEIR 2766 02:05:13,160 --> 02:05:14,400 DISTRIBUTION, IN LIVE HUMAN 2767 02:05:14,400 --> 02:05:14,840 EYES. 2768 02:05:14,840 --> 02:05:16,480 YOU CAN SEE DENSITY OF CELLS 2769 02:05:16,480 --> 02:05:19,640 INCREASES US A MOVE AWAY FROM 2770 02:05:19,640 --> 02:05:20,640 THE FOVEA, CORRELATING WITH 2771 02:05:20,640 --> 02:05:28,040 THICKNESS OF THE NERVE FIBER 2772 02:05:28,040 --> 02:05:29,240 LEVEL WHICH WE THINK THEY HAVE A 2773 02:05:29,240 --> 02:05:31,400 ROLE IN CLEARING AND DISEASES. 2774 02:05:31,400 --> 02:05:34,640 WE'RE ALSO ABLE TO LOOK AT TIME 2775 02:05:34,640 --> 02:05:36,920 LAPSE VIDEOS OF THESE -- AND 2776 02:05:36,920 --> 02:05:38,760 THIS IS A 25-MINUTE TIME LAPSED 2777 02:05:38,760 --> 02:05:41,720 VIDEO OF THE CELLS, YOU CAN SEE 2778 02:05:41,720 --> 02:05:44,480 IN THE RESTING STATE PROBING 2779 02:05:44,480 --> 02:05:45,560 ENVIRONMENTS, THESE IMAGES ARE 2780 02:05:45,560 --> 02:05:48,640 FROM LIVE HUMAN EYES, NOW THAT 2781 02:05:48,640 --> 02:05:50,160 WE CAN EXPLORE THAT, WE 2782 02:05:50,160 --> 02:05:53,240 UNDERTOOK A FEW STUDIES TO LOOK 2783 02:05:53,240 --> 02:05:53,680 AT CHANGES. 2784 02:05:53,680 --> 02:05:56,760 THE FIRST WAS WITH AGING, WE 2785 02:05:56,760 --> 02:06:00,240 FOUND HUMANS LOSE 2% OF 2786 02:06:00,240 --> 02:06:04,080 MICROGLIA PER YEAR, CELLS AREN'T 2787 02:06:04,080 --> 02:06:04,920 REPLENISHED FROM UNDERLYING 2788 02:06:04,920 --> 02:06:05,840 PLEXIFORM LAYERS. 2789 02:06:05,840 --> 02:06:12,720 WE FOUND THE MICRO MICROGLIA AE 2790 02:06:12,720 --> 02:06:16,280 DENSER IN EARLIER STAGES OF 2791 02:06:16,280 --> 02:06:18,320 DISEASE, SLIGHT INCREASE IN 2792 02:06:18,320 --> 02:06:20,200 PROCESS VELOCITY WITH GLAUCOMA. 2793 02:06:20,200 --> 02:06:29,520 ANOTHER STUDY WE RECENTLY 2794 02:06:29,520 --> 02:06:30,920 FINISHED, PUBLISHED, MICROGLIA 2795 02:06:30,920 --> 02:06:33,800 DENSER IN M.S. SUBJECTS THAN 2796 02:06:33,800 --> 02:06:35,720 HEALTHY VOLUNTEERS AND CONFIRMED 2797 02:06:35,720 --> 02:06:37,800 EARLIER WORK THAT THE PROCESS 2798 02:06:37,800 --> 02:06:39,880 VELOCITY IS SLIGHTLY HIGHER WITH 2799 02:06:39,880 --> 02:06:40,120 DISEASE. 2800 02:06:40,120 --> 02:06:44,840 SO IMMUNE CELLS PLAY A ROLE IN 2801 02:06:44,840 --> 02:06:47,240 RETINAL HOMEOSTASIS AND 2802 02:06:47,240 --> 02:06:48,080 NEUROLOGICAL AND OPHTHALMIC 2803 02:06:48,080 --> 02:06:49,720 DISEASE, IMAGING IN LIVING HUMAN 2804 02:06:49,720 --> 02:06:54,440 EYES, EXPLORED IN A NUMBER OF 2805 02:06:54,440 --> 02:06:55,520 DIFFERENT CONDITIONS INCLUDING 2806 02:06:55,520 --> 02:06:58,280 TRAUMATIC BRAIN INJURY, OPTIC 2807 02:06:58,280 --> 02:07:00,160 NEUROPATHY, RETINAL VAIN 2808 02:07:00,160 --> 02:07:01,240 OCCLUSION, DIABETIC RETINOPATHY 2809 02:07:01,240 --> 02:07:02,440 AND OTHERS. 2810 02:07:02,440 --> 02:07:04,200 NOW MOVING FURTHER INTO THE 2811 02:07:04,200 --> 02:07:08,120 RETINA WE'LL LOOK AT RETINAL 2812 02:07:08,120 --> 02:07:09,200 VESSEL AUTOREGULATION IN 2813 02:07:09,200 --> 02:07:09,600 GLAUCOMA. 2814 02:07:09,600 --> 02:07:13,160 SO THE RETINA IS CENTRAL NERVOUS 2815 02:07:13,160 --> 02:07:16,320 SYSTEM TISSUE, IT SHARES 2816 02:07:16,320 --> 02:07:17,680 CHARACTERISTICALLY HIGH 2817 02:07:17,680 --> 02:07:19,080 METABOLISM OF THE BRAIN, 2818 02:07:19,080 --> 02:07:22,000 HYPEROXIA IS A CHANGE IN BLOOD 2819 02:07:22,000 --> 02:07:26,320 OXYGEN PARTIAL PRESSURE THAT 2820 02:07:26,320 --> 02:07:29,320 ACTIVATES AUTOREGULATION, SO OUR 2821 02:07:29,320 --> 02:07:33,160 HYPOTHESIS ABERRANT BLOOD KNOW 2822 02:07:33,160 --> 02:07:34,560 PRECEDES STRUCTURAL CHANGES AND 2823 02:07:34,560 --> 02:07:41,080 CAN BE USED AS BIOMARKER AND TO 2824 02:07:41,080 --> 02:07:46,760 DETERMINE RETINAL VAST VASCULAR 2825 02:07:46,760 --> 02:07:47,200 FUNCTION. 2826 02:07:47,200 --> 02:07:51,680 WE HAVE AN OSLO SCAN, WHICH IS 2827 02:07:51,680 --> 02:07:54,760 SCANNING LASER, IN A LINE SCAN 2828 02:07:54,760 --> 02:07:57,360 MODE WHERE AT TENS OF KILOHERTZ 2829 02:07:57,360 --> 02:07:59,920 RATES TRACK INDIVIDUAL RED BLOOD 2830 02:07:59,920 --> 02:08:02,280 CELLS FLOWING THROUGH VESSELS. 2831 02:08:02,280 --> 02:08:03,800 YOU CAN SEE THE STREAKS, THE 2832 02:08:03,800 --> 02:08:05,480 ANGLE THE STREAK MAKES WITH 2833 02:08:05,480 --> 02:08:09,840 RESPECT TO THE SCANNER AND TO 2834 02:08:09,840 --> 02:08:12,000 VESSEL, GIVES QUANTITATIVE 2835 02:08:12,000 --> 02:08:13,920 MEASURE OF FLOW VELOCITY. 2836 02:08:13,920 --> 02:08:18,640 AND WE CAN PAIR THAT WITH OUR 2837 02:08:18,640 --> 02:08:20,920 ABILITY TO COLLECT VERY HIGH 2838 02:08:20,920 --> 02:08:23,440 RESOLUTION AOOCT VOLUMES WHERE 2839 02:08:23,440 --> 02:08:25,400 WE CAN EXTRACTS VESSEL DIAMETER 2840 02:08:25,400 --> 02:08:27,800 SO YOU CAN SEE HERE, WE CAN 2841 02:08:27,800 --> 02:08:30,200 RESOLVE THE VESSEL WALLS HERE 2842 02:08:30,200 --> 02:08:32,080 AND GET PRECISE MEASURES OF 2843 02:08:32,080 --> 02:08:33,280 VESSEL DIAMETER AND PUTTING 2844 02:08:33,280 --> 02:08:35,120 THOSE TWO TOGETHER ALLOWS US TO 2845 02:08:35,120 --> 02:08:37,400 DO QUANTITATIVE BLOOD FLOW IN 2846 02:08:37,400 --> 02:08:39,040 OUR SUBJECTS. 2847 02:08:39,040 --> 02:08:41,000 SO WE DO AN OXYGEN CHALLENGE 2848 02:08:41,000 --> 02:08:44,400 EXPERIMENT, WHERE WE GIVE THEM 2849 02:08:44,400 --> 02:08:45,720 OXYGEN SUPPLEMENTATION DURING 2850 02:08:45,720 --> 02:08:49,200 IMAGING AND RECORD VITALS AND 2851 02:08:49,200 --> 02:08:49,480 SPO2. 2852 02:08:49,480 --> 02:08:51,840 THE FIRST VALIDATION WE DID OF 2853 02:08:51,840 --> 02:08:53,440 THIS EXPERIMENT WAS A BRANCHED 2854 02:08:53,440 --> 02:08:55,920 VESSEL EXPERIMENT WHERE WE FOUND 2855 02:08:55,920 --> 02:08:57,440 A SUBJECT THAT HAD AN AREA WHERE 2856 02:08:57,440 --> 02:09:00,440 THEY HAD A PARENT AND TWO 2857 02:09:00,440 --> 02:09:01,840 DAUGHTER VESSELS, WE COULD DO 2858 02:09:01,840 --> 02:09:04,800 THE LINE SCAN APPROACH ON THIS 2859 02:09:04,800 --> 02:09:06,680 TO MEASURE THE VELOCITIES, AND 2860 02:09:06,680 --> 02:09:09,640 MEASURE THEIR DIAMETERS, AND WE 2861 02:09:09,640 --> 02:09:10,960 FOUND THAT THE PARENT, MEAN FLOW 2862 02:09:10,960 --> 02:09:12,480 RATE IN THE PARENT, WAS THE SUM 2863 02:09:12,480 --> 02:09:15,320 OF THE TWO DAUGHTER VESSELS TO A 2864 02:09:15,320 --> 02:09:16,600 HIGH DEGREE, WHICH SHOWED 2865 02:09:16,600 --> 02:09:20,120 CONSERVATION OF MEAN FLOW RATES. 2866 02:09:20,120 --> 02:09:21,560 NOW WE'RE TAKING THIS TO A 2867 02:09:21,560 --> 02:09:23,760 HEALTHY COHORT AND THEN TO OUR 2868 02:09:23,760 --> 02:09:29,960 GLAUCOMA SUBJECTS. 2869 02:09:29,960 --> 02:09:35,080 WE CAN SEE CARDIAC CYCLE AND 2870 02:09:35,080 --> 02:09:37,320 PARABOLIC FLOW ACROSS 2871 02:09:37,320 --> 02:09:43,200 VASCULATURE, PUTTING THOSE 2872 02:09:43,200 --> 02:09:46,520 TOGETHER WERE ABLE TO MEASURE, 2873 02:09:46,520 --> 02:09:48,000 ACCOMPANIED WITH 2874 02:09:48,000 --> 02:09:54,160 VASOCONSTRICTION IN ARTERIALS 2875 02:09:54,160 --> 02:09:56,320 AND VENTROLES, NO CHANGE IN MEAN 2876 02:09:56,320 --> 02:10:01,240 FLOW RATES WITH OXYGEN 2877 02:10:01,240 --> 02:10:01,800 SUPPLEMENTATION INDICATING 2878 02:10:01,800 --> 02:10:05,280 EFFICIENT AUTOREGULATION, ONLY 2879 02:10:05,280 --> 02:10:07,280 HAVE DONE ONE GLAUCOMA SUSPECT, 2880 02:10:07,280 --> 02:10:09,680 20% DECREASE IN MEAN FLOW RATE 2881 02:10:09,680 --> 02:10:12,080 SO WE'RE WORKING WITH A FULL 2882 02:10:12,080 --> 02:10:14,320 GLAUCOMA COHORT AND WE SHOULD 2883 02:10:14,320 --> 02:10:15,440 HAVE RESULTS THERE SOON. 2884 02:10:15,440 --> 02:10:18,880 NEXT I'D LIKE TO MOVE TO THE 2885 02:10:18,880 --> 02:10:22,960 PHOTORECEPTOR, NEW FUNCTIONAL 2886 02:10:22,960 --> 02:10:26,320 MEASURE IS CALLED 2887 02:10:26,320 --> 02:10:30,920 OPTORETINOPATHY, COROLLARY OF 2888 02:10:30,920 --> 02:10:33,760 MULTI-FOCAL ERG, TWO VARIETIES 2889 02:10:33,760 --> 02:10:44,280 OF OSLO OR AOOCT BUT MEASURING 2890 02:10:46,520 --> 02:10:49,640 THE SAME THING. 2891 02:10:49,640 --> 02:10:52,480 THE MORE RECENT DEVELOPMENT, YOU 2892 02:10:52,480 --> 02:10:55,840 HOOK AT PHASE CHANGES IN 2893 02:10:55,840 --> 02:10:58,960 SPECIFIC LAYERS OF THE RETINA, 2894 02:10:58,960 --> 02:11:09,440 ENTER SEGMENT, OUTER SEGMENT 2895 02:11:13,080 --> 02:11:22,560 JUNCTION, OPTI RETINOPATHY IS AN 2896 02:11:22,560 --> 02:11:24,320 ESTABLISHED TECHNIQUE. 2897 02:11:24,320 --> 02:11:29,160 A PROCESS WE'RE USING, WE 2898 02:11:29,160 --> 02:11:30,400 COLLECT VOLUMES ACROSS STIMULUS, 2899 02:11:30,400 --> 02:11:32,840 FROM AND REGISTER TO EACH OTHER, 2900 02:11:32,840 --> 02:11:35,360 CAN TRACK PHASE DYNAMICS AND 2901 02:11:35,360 --> 02:11:38,520 INDIVIDUAL CONES, SO AGAIN PHASE 2902 02:11:38,520 --> 02:11:40,480 CHANGE BETWEEN INNER AND OUTER 2903 02:11:40,480 --> 02:11:43,560 SEGMENT AND TIPS WE GET A 2904 02:11:43,560 --> 02:11:46,080 MEASURE OF THE OPTICAL PATH 2905 02:11:46,080 --> 02:11:52,520 LENGTH CHANGES, ELONGATION OF 2906 02:11:52,520 --> 02:11:54,920 PHOTORECEPTORS. 2907 02:11:54,920 --> 02:11:56,560 WE CAN CLASSIFY. 2908 02:11:56,560 --> 02:12:02,040 IF WE IMAGE A REGION WITH NO 2909 02:12:02,040 --> 02:12:02,800 OVERLYING VASCULATURE, APPLY 2910 02:12:02,800 --> 02:12:04,120 STIMULUS, SEE PHASE CHANGE WHICH 2911 02:12:04,120 --> 02:12:08,400 WE CAN EQUATE WITH OPTICAL PATH 2912 02:12:08,400 --> 02:12:10,680 LENGTH CHANGE FROM THE 2913 02:12:10,680 --> 02:12:13,760 650-NANOMETER STIMULUS LIGHT. 2914 02:12:13,760 --> 02:12:15,120 SO, USING THE PRINCIPLE 2915 02:12:15,120 --> 02:12:18,040 COMPONENT ANALYSIS CAN SEPARATE 2916 02:12:18,040 --> 02:12:20,240 L, M, S-CONES IN THIS PHASE 2917 02:12:20,240 --> 02:12:20,680 RESPONSE. 2918 02:12:20,680 --> 02:12:24,080 AND YOU CAN DO THIS WITH HIGH 2919 02:12:24,080 --> 02:12:25,120 ACCURACY, VERY LOW UNCERTAINTY. 2920 02:12:25,120 --> 02:12:28,120 SO THEN YOU CAN START TO APPLY 2921 02:12:28,120 --> 02:12:29,160 OTHER STIMULUS SOURCES. 2922 02:12:29,160 --> 02:12:32,240 WE HAVE A SUPER CONTINUUM LASER 2923 02:12:32,240 --> 02:12:34,880 SOURCE CHANNEL THAT'S COUPLED 2924 02:12:34,880 --> 02:12:40,000 INTO ADAPTIVE OPTICS SYSTEM, AND 2925 02:12:40,000 --> 02:12:42,000 YOU CAN LOOK AT DIFFERENCES IN 2926 02:12:42,000 --> 02:12:44,720 THESE WHERE IF YOU PROBE WITH 2927 02:12:44,720 --> 02:12:48,080 526 LIGHT AS EXPECTED YOU'D SEE 2928 02:12:48,080 --> 02:12:53,360 HIGHER RESPONSE IN M VERSUS L 2929 02:12:53,360 --> 02:12:54,680 CONES FOR 650-NANOMETER LIGHT. 2930 02:12:54,680 --> 02:12:58,320 CAN YOU PLOT THESE ON A 2931 02:12:58,320 --> 02:12:59,400 PRINCIPAL COMPONENT CURVE AND 2932 02:12:59,400 --> 02:13:01,040 SEE CLUSTERING OF INDIVIDUAL 2933 02:13:01,040 --> 02:13:02,480 CONES AND CONE CLASSIFICATION IS 2934 02:13:02,480 --> 02:13:03,880 FINE BUT WHAT WE WANT TO USE 2935 02:13:03,880 --> 02:13:10,000 THIS FOR IS LOOKING AT RETINAL 2936 02:13:10,000 --> 02:13:11,880 DEGENERATION AND CONE 2937 02:13:11,880 --> 02:13:12,520 PHOTORECEPTOR FUNCTION. 2938 02:13:12,520 --> 02:13:15,920 WE CAME ONE A MODEL TO SORT OF 2939 02:13:15,920 --> 02:13:21,720 REPRESENT THIS IN A VECTOR 2940 02:13:21,720 --> 02:13:24,880 REPRESENTATION WHERE THE ANGLE 2941 02:13:24,880 --> 02:13:28,720 FROM THE -- VECTOR FROM ORIGIN 2942 02:13:28,720 --> 02:13:32,120 TO CENTROID OF THOSE M, L, 2943 02:13:32,120 --> 02:13:37,120 S-TYPE CONES GIVE A MEASURE OF 2944 02:13:37,120 --> 02:13:42,160 AMPLITUDE, AND ANGLE IS SPECTRAL 2945 02:13:42,160 --> 02:13:42,440 RESPONSE. 2946 02:13:42,440 --> 02:13:45,680 IF WE LOOK AT A SUBJECT THAT HAS 2947 02:13:45,680 --> 02:13:47,960 HEALTHY EYES, NO DIAGNOSED 2948 02:13:47,960 --> 02:13:53,040 RETINAL DISEASE, AND LOOK AT 2949 02:13:53,040 --> 02:13:58,360 AREAS OF DRUSEN, YOU CAN SEE ALL 2950 02:13:58,360 --> 02:14:05,920 THE WAY ACROSS THE MACULA, NICE 2951 02:14:05,920 --> 02:14:08,520 RESOLUTION OF PHOTORECEPTORS, IN 2952 02:14:08,520 --> 02:14:13,760 DRUSEN LOOK AT CROSS-SECTIONAL B 2953 02:14:13,760 --> 02:14:15,760 SCAN, DRUSEN IS EVIDENT IN BOTH 2954 02:14:15,760 --> 02:14:16,760 VIEWS, WORKING WITH DUKE 2955 02:14:16,760 --> 02:14:19,360 UNIVERSITY ON A MACHINE LEARNING 2956 02:14:19,360 --> 02:14:22,320 ALGORITHM TO MEASURE THE CONE 2957 02:14:22,320 --> 02:14:23,640 OUTER SEGMENT LENGTHS, HAVE DONE 2958 02:14:23,640 --> 02:14:25,280 THAT AS WELL, BUT THE IMPORTANT 2959 02:14:25,280 --> 02:14:28,120 THING IS THE CONES ABOVE THE 2960 02:14:28,120 --> 02:14:29,320 DRUSEN RETAIN THEIR PAIRED 2961 02:14:29,320 --> 02:14:30,880 REFLECTION SO WE'RE ABLE TO 2962 02:14:30,880 --> 02:14:33,200 MEASURE PHASE CHANGES IN THESE 2963 02:14:33,200 --> 02:14:34,080 DRUSEN REGIONS. 2964 02:14:34,080 --> 02:14:39,120 IF WE LOOK IN THE VOLUME AT RPE 2965 02:14:39,120 --> 02:14:40,640 LAYER, WE CAN SEE REGIONS HERE 2966 02:14:40,640 --> 02:14:43,120 WHERE THE RPE CELLS ARE 2967 02:14:43,120 --> 02:14:45,760 RETAINED, BUT THE MAJORITY OF 2968 02:14:45,760 --> 02:14:50,040 THE DRUSEN AREA HAS THIS 2969 02:14:50,040 --> 02:14:51,240 HETEROGENEOUS SCATTERING 2970 02:14:51,240 --> 02:14:51,520 FEATURE. 2971 02:14:51,520 --> 02:14:58,880 AND WE CAN ALSO LOOK AT THE 2972 02:14:58,880 --> 02:15:00,240 CHORIOCAPILLARIS, YOU CAN SEE IN 2973 02:15:00,240 --> 02:15:03,280 THE TEMPORAL PART OF THE MACULA, 2974 02:15:03,280 --> 02:15:04,480 RELATIVELY HEALTHY NORMAL 2975 02:15:04,480 --> 02:15:06,480 VASCULATURE AND CLOSER TO THE 2976 02:15:06,480 --> 02:15:09,080 NASAL REGION YOU SEE DISRUPTION, 2977 02:15:09,080 --> 02:15:10,400 MOST IMPORTANTLY IN THE REGION 2978 02:15:10,400 --> 02:15:13,240 AROUND THE DRUSEN YOU CAN ALSO 2979 02:15:13,240 --> 02:15:17,520 SEE DISRUPTION OF THE RETINAL 2980 02:15:17,520 --> 02:15:19,280 VASCULAR NETWORK THAT SUPPLIES 2981 02:15:19,280 --> 02:15:21,160 PHOTORECEPTORS AND RPE. 2982 02:15:21,160 --> 02:15:24,200 LOOKING AT THE CHANGES IN THESE 2983 02:15:24,200 --> 02:15:25,400 LOCATIONS NEAR THE DRUSEN, THIS 2984 02:15:25,400 --> 02:15:28,000 IS THE FIRST LOCATION, AND IF 2985 02:15:28,000 --> 02:15:31,200 YOU LOOK AT THE ADJACENT 2986 02:15:31,200 --> 02:15:32,280 NON-DRUSEN REGION YOU CAN PLOT 2987 02:15:32,280 --> 02:15:35,560 OUT THE VECTORS AND THEN WHEN 2988 02:15:35,560 --> 02:15:37,680 YOU APPLY OUR ANALYSIS AND LOOK 2989 02:15:37,680 --> 02:15:39,080 AT THE REGIONS AND FURTHER APPLY 2990 02:15:39,080 --> 02:15:41,320 THE VECTORS THERE YOU SEE 2991 02:15:41,320 --> 02:15:43,680 THERE'S A TENDENCY OF A WEAKER 2992 02:15:43,680 --> 02:15:47,520 RESPONSE IN THE DRUSEN REGION 2993 02:15:47,520 --> 02:15:49,360 AND ALSO SMALLER ANGLE, AND THIS 2994 02:15:49,360 --> 02:15:51,960 MAY INDICATE THERE'S A SPECTRAL 2995 02:15:51,960 --> 02:15:52,640 SENSITIVITY SHIFT THAT'S CAUSED 2996 02:15:52,640 --> 02:15:55,080 BY A CHANGE IN THE WAVE GUIDING 2997 02:15:55,080 --> 02:15:57,160 PROPERTIES IN THESE CELLS. 2998 02:15:57,160 --> 02:15:58,440 WE'VE DONE THIS FOR ANOTHER 2999 02:15:58,440 --> 02:16:05,240 REGION IN THE SAME EYE, AND THEN 3000 02:16:05,240 --> 02:16:07,640 IN A FEW SUBJECTS FOR REGIONS 3001 02:16:07,640 --> 02:16:10,040 WE'VE SEEN SIMILAR RESULTS, 3002 02:16:10,040 --> 02:16:12,920 WEAKER RESPONSE IN DRUSEN 3003 02:16:12,920 --> 02:16:15,960 REGION, MEASURABLE REDSHIFT IN 3004 02:16:15,960 --> 02:16:19,240 THE DRUSEN REASON, SO WE THINK 3005 02:16:19,240 --> 02:16:20,760 WE'RE UNDERWAY EXPLORING THIS IN 3006 02:16:20,760 --> 02:16:22,320 ADDITIONAL SUBJECTS WITH DRUSEN 3007 02:16:22,320 --> 02:16:26,040 AND WE THINK THIS IS A SUITABLE 3008 02:16:26,040 --> 02:16:28,440 FOR A BIOMARKER OF EARLY AMD. 3009 02:16:28,440 --> 02:16:31,200 LET'S MOVE INTO THE RPE CELLS 3010 02:16:31,200 --> 02:16:34,000 AND LOOK AT ORGANELLE MOTILITY. 3011 02:16:34,000 --> 02:16:39,200 FIRST WITH OUR ADAPTIVE OPTICS 3012 02:16:39,200 --> 02:16:41,680 SYSTEMS WE'RE ABLE TO DO 3013 02:16:41,680 --> 02:16:44,960 HIGH-SPEED IMAGING AND COLLECT 3014 02:16:44,960 --> 02:16:46,160 VOLUMES ACROSS THE TEMPORAL 3015 02:16:46,160 --> 02:16:48,880 MACULA, IN THIS SUBJECT HERE 3016 02:16:48,880 --> 02:16:51,520 THIS HEALTHY VOLUNTEER, THE 3017 02:16:51,520 --> 02:16:56,520 FOVEA IS HERE, GOING INTO THE 3018 02:16:56,520 --> 02:16:59,280 TEMPORAL MACULA, YOU CAN SEE 3019 02:16:59,280 --> 02:17:10,160 PHOTORECEPTOR CHANGES US A RECEN 3020 02:17:12,880 --> 02:17:14,920 THE PERIPHERY, ALSO THE DOTS, 3021 02:17:14,920 --> 02:17:16,640 ABLE TO QUANTIFY CONE DENSITY 3022 02:17:16,640 --> 02:17:18,080 AND CAN YOU DO THE SAME THING 3023 02:17:18,080 --> 02:17:20,160 WITH RPE CELLS SO WE GET A NICE 3024 02:17:20,160 --> 02:17:25,200 PICTURE OF THE RPE CELLS ACROSS 3025 02:17:25,200 --> 02:17:30,320 THE MACULA AND FOUND THAT THEIR 3026 02:17:30,320 --> 02:17:33,400 DENSITY CHANGES THOUGH NOT AS 3027 02:17:33,400 --> 02:17:34,600 MUCH AS CONE PHOTORECEPTORS, THE 3028 02:17:34,600 --> 02:17:38,320 RATIO OF CELLS THAT SERVES EACH 3029 02:17:38,320 --> 02:17:39,000 CONE PHOTORECEPTOR GOES DOWN 3030 02:17:39,000 --> 02:17:46,760 APPRECIATABLY AS YOU MOVE IN 3031 02:17:46,760 --> 02:17:47,200 ECENTRICITY. 3032 02:17:47,200 --> 02:17:50,040 IN THESE RPE CELLS, THERE'S 3033 02:17:50,040 --> 02:17:50,400 MITOCHONDRIA. 3034 02:17:50,400 --> 02:17:53,080 MELANOSOMES MOVING AROUND, A 3035 02:17:53,080 --> 02:17:54,080 SOURCE OF CONTRAST. 3036 02:17:54,080 --> 02:17:56,640 LOOKING AT INDIVIDUAL SINGLE 3037 02:17:56,640 --> 02:17:58,360 VOLUME FROM PHOTORECEPTORS CAN 3038 02:17:58,360 --> 02:17:59,800 SEE PHOTORECEPTOR LAYER BUT IN 3039 02:17:59,800 --> 02:18:03,320 THE RPE CELLS WE JUST SEE THIS 3040 02:18:03,320 --> 02:18:04,520 SPECKLE NOISE, BUT WHAT WE CAN 3041 02:18:04,520 --> 02:18:06,560 DO IS LOOK AT THIS OVER TIME AND 3042 02:18:06,560 --> 02:18:11,800 THEN WE GET A VERY CONSTANT 3043 02:18:11,800 --> 02:18:14,000 PHOTORECEPTOR PICTURE BUT THIS 3044 02:18:14,000 --> 02:18:16,240 BOILING OF THE TISSUE, NOT 3045 02:18:16,240 --> 02:18:18,280 REALLY BOILING BUT LOOKS LIKE 3046 02:18:18,280 --> 02:18:19,160 IT'S BOILING, DECORRELATED 3047 02:18:19,160 --> 02:18:20,720 LAYINGS IN SPECKLE PATTERN WHICH 3048 02:18:20,720 --> 02:18:23,800 WE CAN USE TO CREATE AVERAGE 3049 02:18:23,800 --> 02:18:34,440 IMAGES, DECOR DECOR -- DECORRE, 3050 02:18:35,120 --> 02:18:39,080 LOOKING AT INDIVIDUAL CELL OVER 3051 02:18:39,080 --> 02:18:43,440 TIME, WE'RE ABLE TO CALCULATE 3052 02:18:43,440 --> 02:18:44,600 THE CROSS-CORRELATION FUNCTION, 3053 02:18:44,600 --> 02:18:45,560 COMPARED TO LAYERS THAT ARE MORE 3054 02:18:45,560 --> 02:18:48,160 STABLE OR THAT DON'T HAVE ANY 3055 02:18:48,160 --> 02:18:48,800 DE-CORRELATION. 3056 02:18:48,800 --> 02:18:51,440 WHEN WE DO THIS WE GET A 3057 02:18:51,440 --> 02:18:52,440 CROSS-CORRELATION COEFFICIENT 3058 02:18:52,440 --> 02:18:56,440 THAT WE CAN FIT TO EXPONENTIAL 3059 02:18:56,440 --> 02:18:59,200 CURVE AND TIME CONSTANT OF WHICH 3060 02:18:59,200 --> 02:19:01,480 IS THE BIOMARKER, MOTILITY TIME 3061 02:19:01,480 --> 02:19:03,040 CONSTANT, AND SEVEN SUBJECTS AND 3062 02:19:03,040 --> 02:19:04,480 EARLY STUDY WE FOUND TIME 3063 02:19:04,480 --> 02:19:06,040 CONSTANT WAS ABOUT FIVE SECONDS. 3064 02:19:06,040 --> 02:19:07,880 AND THAT ALSO PUTS A LIMIT ON 3065 02:19:07,880 --> 02:19:10,360 HOW WE DO OUR IMAGING, WE HAVE 3066 02:19:10,360 --> 02:19:13,000 TO SEPARATE OUR VOLUMES BY FIVE 3067 02:19:13,000 --> 02:19:14,760 SECONDS IN ORDER TO ALLOW 3068 02:19:14,760 --> 02:19:18,240 DE-CORRELATION TO OCCUR TO GET 3069 02:19:18,240 --> 02:19:21,120 THE RPE-ENHANCED CONTRAST. 3070 02:19:21,120 --> 02:19:25,600 SO, WE'RE USING THIS TECHNIQUE 3071 02:19:25,600 --> 02:19:28,080 IN SOME KATHY KUKRAS' SUBJECTS 3072 02:19:28,080 --> 02:19:31,480 BUT IN INITIAL STUDY IN HEALTHY 3073 02:19:31,480 --> 02:19:33,800 SUBJECTS WANTED TO EXPLORE 3074 02:19:33,800 --> 02:19:34,680 SOURCE OF ORGANELLE 3075 02:19:34,680 --> 02:19:38,160 DE-CORRELATION MORE AND LOOKED, 3076 02:19:38,160 --> 02:19:40,040 WITH THE VERY HIGH CAPABILITIES 3077 02:19:40,040 --> 02:19:44,880 OF OCT LOOK AT APICAL, MEDIAL, 3078 02:19:44,880 --> 02:19:46,840 BASAL REGIONS, YOU CAN SEE THESE 3079 02:19:46,840 --> 02:19:48,160 REGIONS SEPARATED BY ABOUT 4 3080 02:19:48,160 --> 02:19:50,560 MICRONS, IF YOU DO AN AVERAGE OF 3081 02:19:50,560 --> 02:19:53,400 THESE VOLUMES YOU SEE THE RPE 3082 02:19:53,400 --> 02:19:54,600 CELLS ARE CLEARLY DELINEATED, 3083 02:19:54,600 --> 02:19:58,440 BUT IF YOU LOOK AT SINGLE 3084 02:19:58,440 --> 02:19:59,240 VOLUMES THERE'S DIFFERENT 3085 02:19:59,240 --> 02:20:00,840 SPECKLE PATTERN IN EACH OF THEM 3086 02:20:00,840 --> 02:20:03,400 AND COMPARING THAT TO THE SIGNAL 3087 02:20:03,400 --> 02:20:05,120 FROM BROOKS MEMBRANE YOU CAN SEE 3088 02:20:05,120 --> 02:20:05,880 THE DIFFERENCES THERE. 3089 02:20:05,880 --> 02:20:08,800 WHEN WE DID THIS AND DID OUR 3090 02:20:08,800 --> 02:20:16,200 MEASUREMENT WE FOUND IN APICAL 3091 02:20:16,200 --> 02:20:20,160 REGION WE GET LONGER TIME 3092 02:20:20,160 --> 02:20:22,760 CONSTANT, IN BASAL REGION WHERE 3093 02:20:22,760 --> 02:20:28,440 THERE'S MITOCHONDRIA WE GET MUCH 3094 02:20:28,440 --> 02:20:30,080 SHORTER TIME CONSTANT, MORE 3095 02:20:30,080 --> 02:20:33,840 ACTIVITY IN THE MITOCHONDRIA AND 3096 02:20:33,840 --> 02:20:36,880 BASAL REGION. 3097 02:20:36,880 --> 02:20:38,960 AND THAT'S JUST HISTOOF THE RPE 3098 02:20:38,960 --> 02:20:41,880 CELL, YOU CAN SEE THE 3099 02:20:41,880 --> 02:20:50,800 MELANOSOMES, NUKE NUCLEUS, 3100 02:20:50,800 --> 02:20:51,760 MITOCHONDRIA. 3101 02:20:51,760 --> 02:20:57,360 WE'LL MOVE TO THE 3102 02:20:57,360 --> 02:20:58,200 CHORIOCAPILLARIS. 3103 02:20:58,200 --> 02:21:00,320 THIS IS A COLLECTION OF SINGLE 3104 02:21:00,320 --> 02:21:02,200 FOLLOWS AND COLLECTED IN 10 3105 02:21:02,200 --> 02:21:12,720 MINUTES, THIS IS AN AOOCTA SCAN 3106 02:21:15,480 --> 02:21:19,120 FROM HEALTHY VOLUNTEER, WE CAN 3107 02:21:19,120 --> 02:21:20,520 RESOLVE THE CHORIOCAPILLARIS, A 3108 02:21:20,520 --> 02:21:23,160 DENSE CAPILLARY NETWORK JUST 3109 02:21:23,160 --> 02:21:25,160 BELOW THE PHOTORECEPTORS. 3110 02:21:25,160 --> 02:21:29,400 AND THEN YOU CAN TAKE A LOOK AT 3111 02:21:29,400 --> 02:21:35,200 THE VASCULAR NETWORK ACROSS THIS 3112 02:21:35,200 --> 02:21:35,960 REGION, NICE DELINEATION OF 3113 02:21:35,960 --> 02:21:37,600 VOIDS AND VESSELS. 3114 02:21:37,600 --> 02:21:39,600 WE STARTED TO EXPLORE THIS IN 3115 02:21:39,600 --> 02:21:41,240 KATHY'S SUBJECTS, THIS IS A 3116 02:21:41,240 --> 02:21:43,720 SUBJECT WITH CONE ROD DYSTROPHY. 3117 02:21:43,720 --> 02:21:46,480 YOU CAN SEE THEY HAVE A CENTRAL 3118 02:21:46,480 --> 02:21:49,080 LESION OF ABOUT 7 OR 8 DEGREES 3119 02:21:49,080 --> 02:21:51,280 WHERE THERE'S A DISRUPTION IN 3120 02:21:51,280 --> 02:21:53,560 THE PHOTORECEPTOR AND RPE CELLS. 3121 02:21:53,560 --> 02:21:57,120 THE PATIENT INTERESTINGLY HAD 3122 02:21:57,120 --> 02:21:58,600 VERY GOOD CENTRAL VISUAL ACUITY. 3123 02:21:58,600 --> 02:22:02,640 SO IF WE TAKE A LOOK AND DO THE 3124 02:22:02,640 --> 02:22:04,880 OCT SCANS ACROSS THE MACULA IN 3125 02:22:04,880 --> 02:22:07,520 THIS SUBJECT, YOU CAN SEE THE 3126 02:22:07,520 --> 02:22:11,400 BROAD OUTLINES OF THE LESION 3127 02:22:11,400 --> 02:22:18,800 HERE, AND IN THE PARAGRAPH RAIL 3128 02:22:18,800 --> 02:22:22,960 D. - PERIPHERAL REGION, WE CAN 3129 02:22:22,960 --> 02:22:23,600 SEE HYPER REFLECTIVE 3130 02:22:23,600 --> 02:22:25,040 PSEUDODRUSEN, WERE ABLE TO 3131 02:22:25,040 --> 02:22:27,720 MEASURE PHOTORECEPTORS IN THE 3132 02:22:27,720 --> 02:22:30,080 CENTRAL LESION WHICH EXPLAINS 3133 02:22:30,080 --> 02:22:32,920 THEIR GOOD VISUAL ACUITIY. 3134 02:22:32,920 --> 02:22:38,720 AND WE CAN APPLY OUR AO-OCTA 3135 02:22:38,720 --> 02:22:41,000 PROCESSING, AND YOU NOTICE 3136 02:22:41,000 --> 02:22:47,520 IMMEDIATELY THE CHANGE IN 3137 02:22:47,520 --> 02:22:50,160 VESSELS IN THE LESION REGION 3138 02:22:50,160 --> 02:22:51,440 ABOUT THINNER CHORIOCAPILLARIS 3139 02:22:51,440 --> 02:22:53,120 VESSELS, UNCLEAR WHETHER THESE 3140 02:22:53,120 --> 02:22:56,600 ARE ACTUALLY THINNER VESSELS OR 3141 02:22:56,600 --> 02:22:58,560 FROM SCATTERING CHANGES. 3142 02:22:58,560 --> 02:23:01,880 WE'VE QUANTIFIED ALL OF THIS AND 3143 02:23:01,880 --> 02:23:05,880 COMPARED THE CONE-ROD DYSTROPHY 3144 02:23:05,880 --> 02:23:14,800 TO HEALTHY VOLUNTEER, FURTHER 3145 02:23:14,800 --> 02:23:15,640 QUANTIFICATION, HEALTHY 3146 02:23:15,640 --> 02:23:16,640 VOLUNTEER DENSITY IS CONSISTENT 3147 02:23:16,640 --> 02:23:20,040 ACROSS THE MACULA, YOU SEE 3148 02:23:20,040 --> 02:23:22,320 CHANGES IN VESSEL DENSITY WHERE 3149 02:23:22,320 --> 02:23:24,440 THE LESION IS IN CONE-ROD 3150 02:23:24,440 --> 02:23:26,680 DYSTROPHY, SIMILARLY WITH THE 3151 02:23:26,680 --> 02:23:30,240 VESSEL DIAMETER THERE MAY BE 3152 02:23:30,240 --> 02:23:33,160 SOME CHANGES IN VESSEL 3153 02:23:33,160 --> 02:23:35,800 TORTUOSITY, AND THEN WE CAN LOOK 3154 02:23:35,800 --> 02:23:40,040 AT REGIONS IN BETWEEN 3155 02:23:40,040 --> 02:23:40,720 VASCULATURE AND SEE CLEAR 3156 02:23:40,720 --> 02:23:47,000 DIFFERENCES IN THE DISEASE EYE 3157 02:23:47,000 --> 02:23:50,800 COMPARED TO HEALTHY EYE. 3158 02:23:50,800 --> 02:23:52,840 HEALTHY VOLUNTEERS HAVE 3159 02:23:52,840 --> 02:23:53,760 CONSISTENT VALUES, CONSISTENT 3160 02:23:53,760 --> 02:23:56,040 VASCULAR NETWORK AND THESE 3161 02:23:56,040 --> 02:23:57,040 CHORIOCAPILLARIS METRICS ARE 3162 02:23:57,040 --> 02:23:58,240 EXTREMELY SENSITIVE TO VASCULAR 3163 02:23:58,240 --> 02:24:01,960 CHANGES SO THEY MAKE VERY GOOD 3164 02:24:01,960 --> 02:24:02,480 BIOMARKERS. 3165 02:24:02,480 --> 02:24:03,680 I'LL STOP THERE AND THE FDA 3166 02:24:03,680 --> 02:24:05,920 MISSION IS TO ENABLE GREATER 3167 02:24:05,920 --> 02:24:07,600 PATIENT ACCESS TO INNOVATIVE 3168 02:24:07,600 --> 02:24:10,280 TECHNOLOGIES WHICH YOU AT THE 3169 02:24:10,280 --> 02:24:13,880 NIH COMPLETELY UNDERSTAND. 3170 02:24:13,880 --> 02:24:14,560 WE'VE ESTABLISHED ADAPTIVE 3171 02:24:14,560 --> 02:24:15,200 OPTICS REGULATORY SCIENCE 3172 02:24:15,200 --> 02:24:16,800 PROGRAM IN OUR OFFICE AND OUR 3173 02:24:16,800 --> 02:24:18,840 LABS ARE EQUIPMENT WITH TWO 3174 02:24:18,840 --> 02:24:22,200 STATE OF THE ART MULTI-MODAL AO 3175 02:24:22,200 --> 02:24:27,240 SYSTEMS THAT SEARCH AS PLATFORM 3176 02:24:27,240 --> 02:24:28,880 FOR CLINICAL STUDIES WITH A 3177 02:24:28,880 --> 02:24:31,160 NUMBER OF COLLABORATORS AT NIH 3178 02:24:31,160 --> 02:24:32,920 NEI, DEMONSTRATING AND DEVELOP 3179 02:24:32,920 --> 02:24:34,240 AO-BASED BIOMARKERS IMPORTANT IN 3180 02:24:34,240 --> 02:24:36,280 VISION SCIENCE STUDIES TO BETTER 3181 02:24:36,280 --> 02:24:37,640 UNDERSTAND RETINAL METABOLISM 3182 02:24:37,640 --> 02:24:40,480 AND FUNCTION, BUT ALSO FOR 3183 02:24:40,480 --> 02:24:43,320 LOOKING AT OPHTHALMIC AND 3184 02:24:43,320 --> 02:24:43,960 NEUROLOGICAL DISEASE DIAGNOSIS 3185 02:24:43,960 --> 02:24:44,520 AND TREATMENT. 3186 02:24:44,520 --> 02:24:54,880 I'LL STOP AND TAKE ANY 3187 02:25:03,640 --> 02:25:03,840 QUESTIONS. 3188 02:25:03,840 --> 02:25:04,120 [APPLAUSE] 3189 02:25:04,120 --> 02:25:06,200 >>HAVE YOU LOOKED AT 3190 02:25:06,200 --> 02:25:07,160 CORRELATION BETWEEN VESSEL 3191 02:25:07,160 --> 02:25:09,840 DIAMETER OR DENSITY AND 3192 02:25:09,840 --> 02:25:10,560 MITOCHONDRIAL FUNCTION? 3193 02:25:10,560 --> 02:25:10,880 >>NOT YET. 3194 02:25:10,880 --> 02:25:12,320 BUT THAT'S A GREAT QUESTION AND 3195 02:25:12,320 --> 02:25:14,440 A GREAT IDEA. 3196 02:25:14,440 --> 02:25:15,400 THAT'S SOMETHING WE WANT TO 3197 02:25:15,400 --> 02:25:16,160 BEGIN TO EXPLORE. 3198 02:25:16,160 --> 02:25:22,400 BUT WE HAVEN'T DONE THAT YET. 3199 02:25:22,400 --> 02:25:25,120 YOU CAN TELL WE'RE ON THE CUSP 3200 02:25:25,120 --> 02:25:26,880 OF GREAT ADVANCES BUT JUST GOT 3201 02:25:26,880 --> 02:25:28,840 ALL THESE TECHNIQUES PERFECTED, 3202 02:25:28,840 --> 02:25:31,680 STILL IN THE INITIATION STAGE 3203 02:25:31,680 --> 02:25:33,200 AND SORT OF EARLY DEVELOPMENT 3204 02:25:33,200 --> 02:25:34,640 STAGE OF THEM, AND THERE'S A LOT 3205 02:25:34,640 --> 02:25:38,640 OF WORK THAT GOES INTO DOING -- 3206 02:25:38,640 --> 02:25:41,320 VALIDATING A BIOMARKER AND DOING 3207 02:25:41,320 --> 02:25:47,280 REPRODUCIBILITY STUDIES BUT 3208 02:25:47,280 --> 02:25:55,920 THAT'S A GREAT IDEA. 3209 02:25:55,920 --> 02:25:56,040 3210 02:25:56,040 --> 02:25:57,440 >> GOING BACK TO THE BEGINNING, 3211 02:25:57,440 --> 02:26:00,840 YOU MENTIONED A NEED FOR 3212 02:26:00,840 --> 02:26:03,440 PHANTOMS, I WONDERED IF YOU ARE 3213 02:26:03,440 --> 02:26:04,000 COMBINING MEASUREMENTS ON 3214 02:26:04,000 --> 02:26:06,000 PHANTOMS WITH ANY OF THESE 3215 02:26:06,000 --> 02:26:07,040 PATIENT MEASUREMENTS OR TO 3216 02:26:07,040 --> 02:26:08,600 CHARACTERIZE THE SYSTEMS AND 3217 02:26:08,600 --> 02:26:10,120 WHAT YOUR NEEDS ARE AROUND 3218 02:26:10,120 --> 02:26:10,320 THOSE. 3219 02:26:10,320 --> 02:26:13,440 >>SO, WE'RE DOING A COUPLE OF 3220 02:26:13,440 --> 02:26:14,560 THINGS. 3221 02:26:14,560 --> 02:26:17,360 ONE, SO THE PHANTOMS THAT WE'RE 3222 02:26:17,360 --> 02:26:18,560 DEVELOPING FIRST LOOKING AT 3223 02:26:18,560 --> 02:26:19,120 RESOLUTION. 3224 02:26:19,120 --> 02:26:21,040 I HAVE A SORT OF A VISION TO 3225 02:26:21,040 --> 02:26:22,720 MAKE THE PHANTOMS A LITTLE MORE 3226 02:26:22,720 --> 02:26:25,800 DYNAMIC AND BE ABLE TO BE LIKE 3227 02:26:25,800 --> 02:26:27,360 STRESS TEST FOR OUR AO SYSTEMS, 3228 02:26:27,360 --> 02:26:29,640 I TALKED ABOUT HOW IT'S A 3229 02:26:29,640 --> 02:26:30,560 PERSONALIZED DEVICE THAT'S 3230 02:26:30,560 --> 02:26:31,280 CHANGING WITH EACH SUBJECT 3231 02:26:31,280 --> 02:26:32,600 THAT'S IN FRONT OF IT. 3232 02:26:32,600 --> 02:26:37,000 RIGHT NOW WE HAVE A NUMBER OF 3233 02:26:37,000 --> 02:26:38,440 RESOLUTION PHANTOMS THAT 3234 02:26:38,440 --> 02:26:39,480 BASICALLY MIMIC THE 3235 02:26:39,480 --> 02:26:42,240 PHOTORECEPTOR DENSITY AND WE'RE 3236 02:26:42,240 --> 02:26:44,440 USING THOSE ON A NUMBER OF 3237 02:26:44,440 --> 02:26:45,720 SYSTEMS INCLUDING THE ONES IN 3238 02:26:45,720 --> 02:26:48,880 JOHNNY'S LAB BUT ALSO IN SOME 3239 02:26:48,880 --> 02:26:50,760 MULTI-SITE CLINICAL STUDIES SO 3240 02:26:50,760 --> 02:26:53,840 THE IDEA THERE TO GET 3241 02:26:53,840 --> 02:26:54,600 STANDARDIZED MEASUREMENTS ACROSS 3242 02:26:54,600 --> 02:26:59,160 ALL OF THE INSTRUMENTS THAT ARE 3243 02:26:59,160 --> 02:27:02,040 IN A MULTI-SITE CLINICAL STUDY, 3244 02:27:02,040 --> 02:27:04,880 IN COLLABORATION WITH JACKIE 3245 02:27:04,880 --> 02:27:07,200 DUNCAN UC-SAN FRANCISCO AND JOE 3246 02:27:07,200 --> 02:27:09,200 CARROLL AT MARQUETTE, SO THEY 3247 02:27:09,200 --> 02:27:10,920 HAVE A LARGE STUDY AT A 3248 02:27:10,920 --> 02:27:12,880 NUMBER -- I THINK FIVE OR SIX 3249 02:27:12,880 --> 02:27:14,760 SITES, WE SEND OUR PHANTOMS 3250 02:27:14,760 --> 02:27:17,160 THERE, THEY TEST THEIR DEVICES, 3251 02:27:17,160 --> 02:27:18,400 MAKE SURE THEY ARE STANDARDIZED 3252 02:27:18,400 --> 02:27:20,240 IN TERMS OF WHAT THE 3253 02:27:20,240 --> 02:27:21,080 MEASUREMENTS AND QUANTIFICATION 3254 02:27:21,080 --> 02:27:23,880 YOU GET OUT OF THEM, AND THEN 3255 02:27:23,880 --> 02:27:25,040 THEY HAVE MORE CONFIDENCE IN THE 3256 02:27:25,040 --> 02:27:26,440 NUMBERS THEY GET FROM THEIR 3257 02:27:26,440 --> 02:27:29,640 CLINICAL STUDIES THAT CAN BE 3258 02:27:29,640 --> 02:27:30,120 COMPARED ACROSS SITES. 3259 02:27:30,120 --> 02:27:31,720 >>WHEN YOU THINK ABOUT DYNAMIC 3260 02:27:31,720 --> 02:27:34,000 ARE YOU THINKING ABOUT SOFT 3261 02:27:34,000 --> 02:27:35,320 MATERIALS YOU CAN STRESS IN REAL 3262 02:27:35,320 --> 02:27:37,000 TIME TO GET THEM TO FLEX? 3263 02:27:37,000 --> 02:27:40,440 >>WELL, THERE'S A NUMBER OF 3264 02:27:40,440 --> 02:27:42,800 DIFFERENT ADAPTIVE LENSES, SO 3265 02:27:42,800 --> 02:27:44,200 IT'S PROBABLY SIMILAR TO WHAT 3266 02:27:44,200 --> 02:27:46,040 YOU'RE TALKING ABOUT, SENSES YOU 3267 02:27:46,040 --> 02:27:49,640 CAN SHAPE AND FOCUS, AND PUTTING 3268 02:27:49,640 --> 02:27:51,720 AN ARBITRARY FOCUS PATTERN OR 3269 02:27:51,720 --> 02:27:53,840 ANY ABERRATIONS ON A LENS AND 3270 02:27:53,840 --> 02:27:55,480 THEN SEEING HOW THE SYSTEM 3271 02:27:55,480 --> 02:27:56,880 RESPONDS TO THAT AND IS ABLE TO 3272 02:27:56,880 --> 02:28:00,080 CORRECT FOR THAT, THAT'S THE 3273 02:28:00,080 --> 02:28:00,280 IDEA. 3274 02:28:00,280 --> 02:28:01,720 >> THANK YOU. 3275 02:28:01,720 --> 02:28:02,560 >> YOU'RE PROBABLY MORE 3276 02:28:02,560 --> 02:28:04,320 FAMILIAR WITH FORMAL LENSES THAN 3277 02:28:04,320 --> 02:28:05,960 I AM, THE MATERIAL THEY USE. 3278 02:28:05,960 --> 02:28:07,560 >> I WAS THINKING ABOUT THE 3279 02:28:07,560 --> 02:28:10,560 PHANTOMS, HOW YOU MAKE A DYNAMIC 3280 02:28:10,560 --> 02:28:11,240 PHANTOMS FOR STRESS TEST. 3281 02:28:11,240 --> 02:28:13,280 >> THERE'S IDEAS WILL YOU TO 3282 02:28:13,280 --> 02:28:15,240 MAKE FLOW PHANTOMS TOO, THAT'S 3283 02:28:15,240 --> 02:28:16,200 MORE DIFFICULT. 3284 02:28:16,200 --> 02:28:17,880 AGAIN, THAT'S SOMETHING WE'RE 3285 02:28:17,880 --> 02:28:25,680 EXPLORING. 3286 02:28:25,680 --> 02:28:26,760 3287 02:28:26,760 --> 02:28:28,520 ALL RIGHT. 3288 02:28:28,520 --> 02:28:29,200 THANK YOU VERY MUCH. 3289 02:28:29,200 --> 02:28:37,480 [APPLAUSE] 3290 02:28:37,480 --> 02:28:45,240 >> DR. MANU PLATT IS DIRECTORY 3291 02:28:45,240 --> 02:28:48,520 OF NIH-WIDE CENTER, ALSO CHIEF 3292 02:28:48,520 --> 02:28:52,920 OF A LAB, I'LL LET HIM DECOMPOSE 3293 02:28:52,920 --> 02:28:53,240 THAT ACRONYM. 3294 02:28:53,240 --> 02:28:54,440 >> THANKS FOR HAVING ME. 3295 02:28:54,440 --> 02:28:56,320 THANKS TO ALL THE ORGANIZERS AND 3296 02:28:56,320 --> 02:28:58,120 THANKS TO MY NEW COLLEAGUES AT 3297 02:28:58,120 --> 02:28:59,040 THE NIH. 3298 02:28:59,040 --> 02:29:01,280 LOOK, MA, I'M PRESENTING AT THE 3299 02:29:01,280 --> 02:29:02,280 NIH! 3300 02:29:02,280 --> 02:29:03,400 NO, IT'S BEEN WONDERFUL GETTING 3301 02:29:03,400 --> 02:29:04,600 THE CENTER OFF THE GROUND AND 3302 02:29:04,600 --> 02:29:05,560 TALKING TO PEOPLE ABOUT WHAT 3303 02:29:05,560 --> 02:29:06,760 WE'RE TRYING TO DO AND WHAT YOU 3304 02:29:06,760 --> 02:29:07,320 ALL DO. 3305 02:29:07,320 --> 02:29:10,080 IT'S GREAT TO COME HOME AND TALK 3306 02:29:10,080 --> 02:29:12,120 ABOUT MY OWN RESEARCH WHICH 3307 02:29:12,120 --> 02:29:13,160 FEELS REALLY GREAT. 3308 02:29:13,160 --> 02:29:15,440 YES, I'M THE DIRECTOR OF THE NEW 3309 02:29:15,440 --> 02:29:26,000 BETA CENTER, RUNNING MY OWN LAP 3310 02:29:28,280 --> 02:29:28,680 HERE. 3311 02:29:28,680 --> 02:29:29,640 MATRICES LAB, THERE'S GREAT 3312 02:29:29,640 --> 02:29:31,440 SICKLE CELL RESEARCH AT NIH. 3313 02:29:31,440 --> 02:29:33,600 TO PLUG MY LAB WE DO MORE THAN 3314 02:29:33,600 --> 02:29:37,080 SICKLE CELL DISEASE BUT I SPEND 3315 02:29:37,080 --> 02:29:40,160 TIME TRYING TO UNDERSTAND WHY 3316 02:29:40,160 --> 02:29:42,920 CHILDREN ARE AT INCREASED RISK 3317 02:29:42,920 --> 02:29:47,280 OF STROKES, BUT WE CARE ABOUT 3318 02:29:47,280 --> 02:29:52,400 THE EXTRACELLULAR MATRIX ME 3319 02:29:52,400 --> 02:29:53,600 MODELING AND PROTEOLYTIC 3320 02:29:53,600 --> 02:29:54,600 CHANGES, TO ALSO UNDERSTAND SOME 3321 02:29:54,600 --> 02:29:56,360 OF THESE THINGS HAPPENING IN 3322 02:29:56,360 --> 02:29:57,440 THESE INDIVIDUALS WITH SICKLE 3323 02:29:57,440 --> 02:30:01,400 CELL, MY LAB IS USING LABEL-FREE 3324 02:30:01,400 --> 02:30:03,160 MAGNETIC RESONANCE IMAGING, BUT 3325 02:30:03,160 --> 02:30:05,240 WE ALSO MERGED THAT WITH 3326 02:30:05,240 --> 02:30:07,640 COMPUTATIONAL FLUID DYNAMICS AND 3327 02:30:07,640 --> 02:30:13,000 HEMO DYNAMICS ALSO LINKED TO 3328 02:30:13,000 --> 02:30:14,120 BIOLOGICAL RESPONSES. 3329 02:30:14,120 --> 02:30:17,360 CHECK IT OUT IF THIS IS 3330 02:30:17,360 --> 02:30:17,720 INTERESTING. 3331 02:30:17,720 --> 02:30:21,880 TODAY WE'LL TALK ABOUT SINGLE 3332 02:30:21,880 --> 02:30:24,960 CELL DISEASE, MULTI-FACTOR 3333 02:30:24,960 --> 02:30:25,360 ZEALS. 3334 02:30:25,360 --> 02:30:26,680 THERE'S ONE NUCLEOTIDE CHANGE 3335 02:30:26,680 --> 02:30:30,720 FROM A TO T, THAT CHANGES ONE 3336 02:30:30,720 --> 02:30:38,160 AMINO ACID, GLUTAMIC TO VALENE, 3337 02:30:38,160 --> 02:30:39,600 HEMOGLOBIN ONLY EXPRESSED IN ONE 3338 02:30:39,600 --> 02:30:41,120 TYPE OF CELL, RED BLOOD CELLS. 3339 02:30:41,120 --> 02:30:42,160 WHAT'S THE PROBLEM? 3340 02:30:42,160 --> 02:30:43,720 RED BLOOD CELLS GO EVERYWHERE. 3341 02:30:43,720 --> 02:30:47,120 AND THAT'S WHERE YOU GET THIS 3342 02:30:47,120 --> 02:30:47,520 SYSTEMIC DAMAGE. 3343 02:30:47,520 --> 02:30:49,880 EVEN TODAY SICKLE CELL IS NOT 3344 02:30:49,880 --> 02:30:54,280 CURED, LIFE EXPECTANCY IS LOWER, 3345 02:30:54,280 --> 02:30:58,160 36 FOR MEN, 42 FOR WOMEN, 3346 02:30:58,160 --> 02:31:01,000 THERE'S EVEN A SEX DIFFERENCE IN 3347 02:31:01,000 --> 02:31:02,120 LIFE EXPECTANCY, AND A HEALTH 3348 02:31:02,120 --> 02:31:04,120 DISPARITY IN THE UNITED STATES. 3349 02:31:04,120 --> 02:31:11,320 ONE IN 400 AFRICAN AMERICANS IS 3350 02:31:11,320 --> 02:31:13,040 HOMOZYGOUS, AND 1 IN 12 ARE 3351 02:31:13,040 --> 02:31:13,400 CARRIERS. 3352 02:31:13,400 --> 02:31:16,120 THERE'S 100,000 AMERICANS IN THE 3353 02:31:16,120 --> 02:31:20,280 U.S. LIVING WITH SICKLE CELL 3354 02:31:20,280 --> 02:31:26,080 DISEASE BUT GLOBAL LIP MILLIONS. 3355 02:31:26,080 --> 02:31:30,760 YOU CAN SEE THAT SUB-SAHARAN 3356 02:31:30,760 --> 02:31:33,560 AFRICA IS MAJOR, INDIAN, ARABIC 3357 02:31:33,560 --> 02:31:36,600 WORLD, NORTH AND SOUTH AMERICA 3358 02:31:36,600 --> 02:31:40,640 AS REMNANTS OF CHATTEL SURGERY. 3359 02:31:40,640 --> 02:31:42,040 90% OF GLOBALLY DIE IN THE FIRST 3360 02:31:42,040 --> 02:31:45,120 FIVE YEARS OF LIFE, MAKING IT A 3361 02:31:45,120 --> 02:31:46,120 DIFFICULT PROBLEM TO QUESTION, 3362 02:31:46,120 --> 02:31:51,360 TUGGING AT MY HEART STRINGS. 3363 02:31:51,360 --> 02:31:53,040 MOLECULAR ORIGINS WE'VE KNOWN 3364 02:31:53,040 --> 02:31:54,760 FOR YEARS, IDENTIFIED CLINICALLY 3365 02:31:54,760 --> 02:31:57,600 IN 1910, WE'VE BEEN STUDYING 3366 02:31:57,600 --> 02:32:03,520 THIS FOR MORE THAN 113 YEARS. 3367 02:32:03,520 --> 02:32:10,400 FROM THE HYDROPHILIC TO VALENE 3368 02:32:10,400 --> 02:32:12,040 IT EXPOSES THAT VALENE TO 3369 02:32:12,040 --> 02:32:18,720 SURFACE OF MOLECULE, VALENE IS 3370 02:32:18,720 --> 02:32:21,320 HYDROPHOBIC, ALL THE HEMOGLOBINS 3371 02:32:21,320 --> 02:32:24,120 SEQUESTER AND SELF-ASSEMBLE. 3372 02:32:24,120 --> 02:32:26,640 IF YOU'RE A POLYMER CHEMIST, 3373 02:32:26,640 --> 02:32:28,000 GREAT QUESTIONS, CREATING A 3374 02:32:28,000 --> 02:32:29,280 STIFF FIBER THAT DEFORMS THE RED 3375 02:32:29,280 --> 02:32:30,960 CELLS AS YOU CAN SEE IN THE 3376 02:32:30,960 --> 02:32:34,440 VIDEO, WE TOOK IN MY LAB UNDER 3377 02:32:34,440 --> 02:32:36,960 THE OXYGENATION CONDITIONS. 3378 02:32:36,960 --> 02:32:40,600 AND BECAUSE THEY ARE RED BLOOD 3379 02:32:40,600 --> 02:32:46,840 CELLS, EFFECTED NUCLEUS, RIGID 3380 02:32:46,840 --> 02:32:49,320 PART, THIS IS DEFORMABLE. 3381 02:32:49,320 --> 02:32:50,520 DEOXYGENATED CONDITIONS WE SEE 3382 02:32:50,520 --> 02:32:52,080 THIS HAPPENING. 3383 02:32:52,080 --> 02:32:54,680 YOU IN THE CAPILLARIES WHERE THE 3384 02:32:54,680 --> 02:32:56,320 CELLS RELEASE OXYGEN AND START 3385 02:32:56,320 --> 02:33:00,520 TO TAKE ON CO2 YOU BEGIN TO GET 3386 02:33:00,520 --> 02:33:01,160 SICKLING AND THESE 3387 02:33:01,160 --> 02:33:08,800 VASOOCCLUSIONS AND BLOCKAGES IN 3388 02:33:08,800 --> 02:33:15,080 POST-CAPILLARY VENULES, LEADING 3389 02:33:15,080 --> 02:33:16,040 TO CRISIS AND PAIN. 3390 02:33:16,040 --> 02:33:18,560 WHEN I STARTED MY LAB AT GEORGIA 3391 02:33:18,560 --> 02:33:22,920 TECH, I FOUND OUT CHILDREN WITH 3392 02:33:22,920 --> 02:33:23,920 SICKLE CELL WERE AT INCREASED 3393 02:33:23,920 --> 02:33:26,000 RISK OF STROKE. 3394 02:33:26,000 --> 02:33:32,120 MY Ph.D. WAS CARDIAC DISEASE, 3395 02:33:32,120 --> 02:33:33,000 ATHEROSCLEROSIS, IN OXYGENATED 3396 02:33:33,000 --> 02:33:34,720 BLOODS CREST ALSO. 3397 02:33:34,720 --> 02:33:37,800 DEOXYGEN RATED WHEN THE GET RED 3398 02:33:37,800 --> 02:33:40,880 CELL SICKLING, WHAT'S HAPPENING 3399 02:33:40,880 --> 02:33:41,320 HERE? 3400 02:33:41,320 --> 02:33:43,600 THERE ARE STUDIES SHOWING THE 3401 02:33:43,600 --> 02:33:44,920 TIME FOR THE SICKLE POLYMER TO 3402 02:33:44,920 --> 02:33:48,200 MELT IS LONGER THAN THE TIME TO 3403 02:33:48,200 --> 02:33:49,960 TRAVERSE THROUGHOUT THE 3404 02:33:49,960 --> 02:33:50,800 OXYGENATED BLOODSTREAM SO 3405 02:33:50,800 --> 02:33:55,640 STARTED LOOKING AT PUTTING 3406 02:33:55,640 --> 02:33:59,440 TOGETHER SOME INTERESTING 3407 02:33:59,440 --> 02:34:04,000 PARADIGMS ABOUT WHAT SICKLING 3408 02:34:04,000 --> 02:34:06,320 CELLS AND WHAT DAMAGE TO LARGE 3409 02:34:06,320 --> 02:34:06,560 ARTERIES. 3410 02:34:06,560 --> 02:34:08,720 11% OF CHILDREN WILL HAVE A 3411 02:34:08,720 --> 02:34:10,760 STROKE, A MAJOR STROKE, BEFORE 3412 02:34:10,760 --> 02:34:12,560 AGE 16, BUT UP TO A THIRD OF 3413 02:34:12,560 --> 02:34:16,880 THESE WILL HAVE WHAT WE CALL 3414 02:34:16,880 --> 02:34:23,280 SILENT STROKE, IT DOESN'T 3415 02:34:23,280 --> 02:34:24,800 PROGRAM BUT UNDER MR, YOU SEE 3416 02:34:24,800 --> 02:34:26,440 DAMAGE TO THE BRAIN. 3417 02:34:26,440 --> 02:34:28,440 SILENT STROKES AFFECT QUALITY OF 3418 02:34:28,440 --> 02:34:29,480 LIFE, LOWER ACADEMIC ACHIEVEMENT 3419 02:34:29,480 --> 02:34:31,240 FOR CHILDREN WITH SICKLE CELL, 3420 02:34:31,240 --> 02:34:32,360 PARTIALLY BECAUSE OF THE STROKES 3421 02:34:32,360 --> 02:34:37,560 AND DAMAGES TO THEIR LESIONS, 3422 02:34:37,560 --> 02:34:38,600 HELD BEHIND. 3423 02:34:38,600 --> 02:34:40,920 BUT AS THE PATIENTS AGE, AGAIN 3424 02:34:40,920 --> 02:34:43,400 HUGE RISK FOR CHILDREN TO HAVE 3425 02:34:43,400 --> 02:34:46,280 ISCHEMIC STROKE BUT AS THEY AGE 3426 02:34:46,280 --> 02:34:47,760 THEN THE RISK INCREASES FOR THEM 3427 02:34:47,760 --> 02:34:51,720 TO GO FROM ISCHEMIC TO 3428 02:34:51,720 --> 02:34:52,720 HEMORRHAGIC, BY TWENTIES AND 3429 02:34:52,720 --> 02:34:54,600 30s BACK TO INFARCTIVE AGAIN. 3430 02:34:54,600 --> 02:34:59,040 WHAT'S HAPPENING WITH THIS 3431 02:34:59,040 --> 02:35:00,080 AGE-RELATED DIFFERENCES? 3432 02:35:00,080 --> 02:35:07,080 WE START TO EQUATE WITH MOUSE 3433 02:35:07,080 --> 02:35:09,120 AGE FOR INTERVENTIONAL STUDIES. 3434 02:35:09,120 --> 02:35:11,960 MY OLD WORK WAS ENDOTHELIAL 3435 02:35:11,960 --> 02:35:13,600 BIOLOGY, SHEER STRESS, LOOKING 3436 02:35:13,600 --> 02:35:16,920 AT SICKLE CELL THERE WAS AMAZING 3437 02:35:16,920 --> 02:35:21,720 CLINICAL TRIAL CALLED THE STOP 3438 02:35:21,720 --> 02:35:26,320 TRIAL BY BE ON BOB ADAMS, 3439 02:35:26,320 --> 02:35:27,840 TRANSCRANIAL DOPPLER, INTERNAL 3440 02:35:27,840 --> 02:35:30,880 CAROTID ARTERY, MIDDLE CEREBRAL 3441 02:35:30,880 --> 02:35:33,280 AND INTERIOR CEREBRAL ARTERY IF 3442 02:35:33,280 --> 02:35:34,640 GREATER THAN 200 CENTIMETERS PER 3443 02:35:34,640 --> 02:35:35,480 SECOND AT INCREASED RISK OF 3444 02:35:35,480 --> 02:35:37,680 STROKES, PUT THEM ON MONTHLY 3445 02:35:37,680 --> 02:35:39,000 BLOOD TRANSFUSIONS AND THIS IS A 3446 02:35:39,000 --> 02:35:40,280 CLINICAL TRIAL THAT ENDED EARLY 3447 02:35:40,280 --> 02:35:42,280 BECAUSE IT WAS OBVIOUS CHILDREN 3448 02:35:42,280 --> 02:35:43,480 WITH BLOODED TRANSFUSIONS WERE 3449 02:35:43,480 --> 02:35:46,200 BEING PROTECTED AND THOSE 3450 02:35:46,200 --> 02:35:47,440 WITHOUT WERE HAVING STROKES 3451 02:35:47,440 --> 02:35:51,280 AND/OR DYING, A GREAT FINDING. 3452 02:35:51,280 --> 02:35:52,880 YOU CAN IDENTIFY WHICH CHILDREN 3453 02:35:52,880 --> 02:35:56,600 ARE AT GREATEST RISK FOR 3454 02:35:56,600 --> 02:35:57,080 STROKES. 3455 02:35:57,080 --> 02:36:03,640 WELL, IF YOU START TO HAVE 3456 02:36:03,640 --> 02:36:05,800 INCREASED BLOOD FOE YOU CAN HAVE 3457 02:36:05,800 --> 02:36:07,560 STENOSIS, WHY MAY BE THERE 3458 02:36:07,560 --> 02:36:10,440 STENOSIS EXCEPT WHEN THEY WERE 3459 02:36:10,440 --> 02:36:11,800 DOING MAGNETIC RESONANCE 3460 02:36:11,800 --> 02:36:14,080 ANGIOGRAMS YOU DIDN'T ALWAYS 3461 02:36:14,080 --> 02:36:16,920 DETECT STENOSIS WHERE YOU WOULD 3462 02:36:16,920 --> 02:36:17,960 GET INCREASED BLOOD FLOW, LOT OF 3463 02:36:17,960 --> 02:36:20,520 CELLS ABOUT WHAT ARE THE RED 3464 02:36:20,520 --> 02:36:22,080 CELLS DOING, THEY ARE ADHESIVE 3465 02:36:22,080 --> 02:36:26,120 AND CAN STICK TO THE WALL OF THE 3466 02:36:26,120 --> 02:36:30,840 ARTERY, AND MORE TRANSIENT IN A 3467 02:36:30,840 --> 02:36:31,720 LUMINAL REMODELING FOR PLAQUE 3468 02:36:31,720 --> 02:36:33,320 AND SICKLE CELL. 3469 02:36:33,320 --> 02:36:36,640 YOU WOULD GET THIS DOWNSTREAM 3470 02:36:36,640 --> 02:36:40,400 REGION OF FLOW REVERSAL 3471 02:36:40,400 --> 02:36:42,320 EXACERBATING PATHWAYS LEADING TO 3472 02:36:42,320 --> 02:36:43,640 SICKLE CELL CAUSING THIS 3473 02:36:43,640 --> 02:36:44,640 ARTERIAL DAMAGE. 3474 02:36:44,640 --> 02:36:47,360 FOR THOSE THAT DON'T STUDY 3475 02:36:47,360 --> 02:36:48,120 CARDIOVASCULAR AND HEMODYNAMICS, 3476 02:36:48,120 --> 02:36:51,080 PATHOLOGIST WAY BACK IN THE 3477 02:36:51,080 --> 02:36:56,760 '60s IDENTIFIED WHERE PLAQUES 3478 02:36:56,760 --> 02:36:59,320 FORM, REGIONS OF BIFURCATIONS, 3479 02:36:59,320 --> 02:37:00,960 CALLED HIS ENGINEER BUDDY, MY 3480 02:37:00,960 --> 02:37:02,360 FORMER DEAN DOWN AT GEORGIA 3481 02:37:02,360 --> 02:37:04,440 TECH, THEY SHOWED AT THESE 3482 02:37:04,440 --> 02:37:06,200 REGIONS OF BIFURCATION YOU GET 3483 02:37:06,200 --> 02:37:10,880 REGIONS OF FLOW SEPARATION AND 3484 02:37:10,880 --> 02:37:12,440 FLOW REVERSAL WITH CARDIAC 3485 02:37:12,440 --> 02:37:12,640 CYCLE. 3486 02:37:12,640 --> 02:37:18,880 THIS IS WHERE A PLAQUE WOULD 3487 02:37:18,880 --> 02:37:20,760 FORM IN THE ARTERY. 3488 02:37:20,760 --> 02:37:24,160 WHEN YOU GO INTO THE BRAIN, 3489 02:37:24,160 --> 02:37:25,280 INTERNAL CAROTID ARTERY BRANCHES 3490 02:37:25,280 --> 02:37:27,000 LIKE CRAZY INSIDE THE BRAIN, 3491 02:37:27,000 --> 02:37:29,200 THERE'S THESE POINTS WHERE YOU 3492 02:37:29,200 --> 02:37:37,720 COULD HAVE REGIONS OF BLOOD 3493 02:37:37,720 --> 02:37:39,800 FLOW. 3494 02:37:39,800 --> 02:37:45,720 IN AUTOPSY, IN THIS 5-YEAR-OLD 3495 02:37:45,720 --> 02:37:51,080 YOU CAN SEE THE LUMINA IS CLOSED 3496 02:37:51,080 --> 02:37:52,200 OFF, FRAGMENTED ELASTIC LAMINA, 3497 02:37:52,200 --> 02:37:53,280 DAMAGED LAYERS. 3498 02:37:53,280 --> 02:37:58,280 WHICH WE KNOW WHEN YOU BREAK 3499 02:37:58,280 --> 02:38:01,360 ELASTIN THEY CHANGE PHENOTYPE, 3500 02:38:01,360 --> 02:38:02,240 BECOME PROLIFERATIVE, ALL THESE 3501 02:38:02,240 --> 02:38:03,320 THINGS COULD HAPPEN DUE TO 3502 02:38:03,320 --> 02:38:05,760 DAMAGE TO THE EXTRACELLULAR 3503 02:38:05,760 --> 02:38:06,600 MATRIX. 3504 02:38:06,600 --> 02:38:17,160 I LOVE THESE ENZYMES, PROTEASES. 3505 02:38:25,840 --> 02:38:26,880 CYSTEINE CATHEPSINS, THE ARTERY 3506 02:38:26,880 --> 02:38:29,480 WANTS TO KEEP THAT LUMEN OPEN AS 3507 02:38:29,480 --> 02:38:31,120 LONG AS POSSIBLE UNTIL THAT 3508 02:38:31,120 --> 02:38:32,760 PLAQUE ENCROACHES TOO FAR. 3509 02:38:32,760 --> 02:38:37,800 WE START LOOKING AT CATHEPSINS, 3510 02:38:37,800 --> 02:38:39,640 REGULATED BY SHEER STRESS 3511 02:38:39,640 --> 02:38:41,080 PUTTING ON FLOW REMODELING 3512 02:38:41,080 --> 02:38:42,480 ISSUES HAPPENING IN SICKLE CELL 3513 02:38:42,480 --> 02:38:44,800 DISEASE, AND WE GOT A STUDY, 3514 02:38:44,800 --> 02:38:45,000 RIGHT? 3515 02:38:45,000 --> 02:38:49,640 A LOT OF BIOMEDICAL FRIENDS 3516 02:38:49,640 --> 02:38:50,960 STUDY MATRIX PROTEINASES, I'M 3517 02:38:50,960 --> 02:38:53,880 HAPPY FOR YOU, GREAT WORK THAT'S 3518 02:38:53,880 --> 02:38:55,960 BEEN HAPPENING, BUT THE CYSTEINE 3519 02:38:55,960 --> 02:38:57,840 CATHEPSINS ARE MORE POWERFUL AS 3520 02:38:57,840 --> 02:39:00,560 THEY TURN OVER SUBSTRATES FASTER 3521 02:39:00,560 --> 02:39:03,080 AND ARE TURNED OFF OR LOW LEVELS 3522 02:39:03,080 --> 02:39:07,360 BUT GET TURNED ON AND UP UNDER 3523 02:39:07,360 --> 02:39:08,440 DISEASE. 3524 02:39:08,440 --> 02:39:19,120 BIG PHARMA HAS BEEN CHASING 3525 02:39:19,120 --> 02:39:21,680 INHIBITORS, BUT 16 MADE IT, 16 3526 02:39:21,680 --> 02:39:22,000 FAILED. 3527 02:39:22,000 --> 02:39:24,400 WE'RE STILL LOOKING FOR DRUGS TO 3528 02:39:24,400 --> 02:39:26,400 INHIBIT THESE ENZYMES TO STOP 3529 02:39:26,400 --> 02:39:26,760 THAT REMODELING. 3530 02:39:26,760 --> 02:39:28,800 ALL OF THAT IS BACKGROUND TO 3531 02:39:28,800 --> 02:39:30,520 SHOW WE WERE TAKING THIS 3532 02:39:30,520 --> 02:39:33,080 MULTI-SCALE APPROACH WHERE WE 3533 02:39:33,080 --> 02:39:34,360 CARE ABOUT ENZYMES BEING 3534 02:39:34,360 --> 02:39:36,680 ELEVATED, HOW ARE THEY BEING 3535 02:39:36,680 --> 02:39:38,280 INFLUENCED, AS WELL AS 3536 02:39:38,280 --> 02:39:39,400 INFLAMMATION IN THE BIOCHEMISTRY 3537 02:39:39,400 --> 02:39:41,360 THAT LEADS TO THEIR 3538 02:39:41,360 --> 02:39:42,240 UPREGULATION, THAT WILL REMODEL 3539 02:39:42,240 --> 02:39:44,520 THAT ARTERY THAT COULD LEAD TO 3540 02:39:44,520 --> 02:39:45,960 THIS PATHOPHYSIOLOGY OF A 3541 02:39:45,960 --> 02:39:46,200 STROKE. 3542 02:39:46,200 --> 02:39:48,160 HOPEFULLY WE FIND OTHER 3543 02:39:48,160 --> 02:39:49,920 BIOMARKERS ALONG THE WAY FOR 3544 02:39:49,920 --> 02:39:52,880 OTHER DRUG TARGETS. 3545 02:39:52,880 --> 02:39:54,400 SO, WORKING WITH RUSSELL AT 3546 02:39:54,400 --> 02:39:56,680 SAINT ST. 3547 02:39:56,680 --> 02:39:59,320 JUDE'S MY FORMER STUDENT WAS 3548 02:39:59,320 --> 02:40:01,080 ABLE TO TAKE ANGIOGRAMS FROM 3549 02:40:01,080 --> 02:40:04,040 CHILDREN WITH SICKLE CELL, 3550 02:40:04,040 --> 02:40:05,480 RECREATED IN SILICO, 3551 02:40:05,480 --> 02:40:06,240 COMPUTATIONAL FLUID DYNAMICS TO 3552 02:40:06,240 --> 02:40:10,920 LOOK AT WHERE ARE THESE POSSIBLE 3553 02:40:10,920 --> 02:40:14,960 REGIONS OF STENOSIS OR 3554 02:40:14,960 --> 02:40:16,400 FLOW-MEDIATED CHANGES CAUSING 3555 02:40:16,400 --> 02:40:18,480 INCREASED BLOOD VELOCITIES, IT 3556 02:40:18,480 --> 02:40:19,240 WASN'T OBVIOUS. 3557 02:40:19,240 --> 02:40:22,640 AND IN DOING SO WE IDENTIFIED 3558 02:40:22,640 --> 02:40:25,400 THAT THE GEOMETRY ALONE, ANATOMY 3559 02:40:25,400 --> 02:40:35,840 OF THE CHILDREN IS QUITE 3560 02:40:39,160 --> 02:40:39,360 DIFFERENT. 3561 02:40:39,360 --> 02:40:40,880 YOU CAN SEE THE SAME INPUT IN 3562 02:40:40,880 --> 02:40:46,640 THE ICA BUT IN THE SICKLE 3563 02:40:46,640 --> 02:40:49,160 VESSELS MCA, IT'S GOING UP. 3564 02:40:49,160 --> 02:40:51,120 THERE'S THESE BUMPS IN THE 3565 02:40:51,120 --> 02:40:54,640 VESSELS, WE CALL THEM BUMPS, 3566 02:40:54,640 --> 02:40:56,520 MICROFEATURES TO MAKE IT SEEM 3567 02:40:56,520 --> 02:40:59,000 FANCY, YOU CAN IMAGINE FOR 3568 02:40:59,000 --> 02:41:00,320 MAGNETIC RESONANCE ANGIOGRAM 3569 02:41:00,320 --> 02:41:01,640 THIS IS THE NEGATIVE OF THE 3570 02:41:01,640 --> 02:41:02,400 BLOOD VESSEL. 3571 02:41:02,400 --> 02:41:03,920 I WAS UPSET. 3572 02:41:03,920 --> 02:41:05,920 CHRIS, DID YOU NOT SMOOTH IT? 3573 02:41:05,920 --> 02:41:07,880 HE FOLLOWED THE SAME ALGORITHMS 3574 02:41:07,880 --> 02:41:09,920 FOR NON-SICKLE AND SICKLE BUT 3575 02:41:09,920 --> 02:41:11,480 WHEN WE THEN LOOKED AT REGIONS 3576 02:41:11,480 --> 02:41:13,520 OF FLOW SEPARATION AND FLOW 3577 02:41:13,520 --> 02:41:15,720 CIRCULATION WE SAW IN THE SICKLE 3578 02:41:15,720 --> 02:41:16,600 CELL INDIVIDUALS, FROM HUMANS, 3579 02:41:16,600 --> 02:41:18,480 AGAIN THOSE ARE THE SITES WHERE 3580 02:41:18,480 --> 02:41:22,400 WE ACTUALLY SAW REGIONS OF FLOW 3581 02:41:22,400 --> 02:41:23,120 SEPARATION AND RECIRCULATION 3582 02:41:23,120 --> 02:41:24,640 WITH THE CARDIAC CYCLE AND DID 3583 02:41:24,640 --> 02:41:26,560 NOT SEE THAT IN INDIVIDUALS 3584 02:41:26,560 --> 02:41:29,920 WITHOUT SICKLE CELL AT THAT 3585 02:41:29,920 --> 02:41:35,440 CRITICAL JUNCTION. 3586 02:41:35,440 --> 02:41:38,080 THE ANATOMY IS DIFFERENT, WE 3587 02:41:38,080 --> 02:41:39,160 CARE ABOUT SUBSTRUCTURAL 3588 02:41:39,160 --> 02:41:39,840 CHANGES. 3589 02:41:39,840 --> 02:41:42,880 WE CAN'T LOOK IN HUMANS, I 3590 02:41:42,880 --> 02:41:45,640 APPLAUD ALL OF YOU, BUT 3591 02:41:45,640 --> 02:41:48,240 UNIVERSITY OF ALABAMA BIRMINGHAM 3592 02:41:48,240 --> 02:41:52,520 DEVELOPED THIS SICKLE CELL 3593 02:41:52,520 --> 02:41:57,200 TOWNES TRANSGENIC MOUSE, KNOCKED 3594 02:41:57,200 --> 02:41:58,920 OUT HOUSE HEMOGLOBIN AND KNOCKED 3595 02:41:58,920 --> 02:42:06,760 IN THE HUMAN, FOLLOWING 3596 02:42:06,760 --> 02:42:11,200 MENDELIAN GENT IS. 3597 02:42:11,200 --> 02:42:17,240 SS IS HOMOZYGOUS. 3598 02:42:17,240 --> 02:42:19,080 YOU'LL SEE WE NORMALLY COMPARE 3599 02:42:19,080 --> 02:42:20,880 WITH AS FOR CONTROL BECAUSE 3600 02:42:20,880 --> 02:42:22,480 THESE ARE LITTER MATE CONTROLS. 3601 02:42:22,480 --> 02:42:26,400 THE FIRST THING WE WANTED TO DO 3602 02:42:26,400 --> 02:42:28,200 EARLY ARTERIAL DAMAGE TO THE 3603 02:42:28,200 --> 02:42:30,080 CAROTID ARTERIES IN THESE MICE, 3604 02:42:30,080 --> 02:42:31,920 WE DID CAROTID BECAUSE MICE ARE 3605 02:42:31,920 --> 02:42:34,440 SMALL, SO RIGHT AT ONE MONTH AT 3606 02:42:34,440 --> 02:42:41,920 WEANING WE ISOLATED CAROTID 3607 02:42:41,920 --> 02:42:44,520 ARTERIES, WE WERE ASTOUNDED, 3608 02:42:44,520 --> 02:42:45,920 ALREADY SEEING A NUMBER OF 3609 02:42:45,920 --> 02:42:50,280 ELASTIN BREAKS BY THREE WEEKS, 3610 02:42:50,280 --> 02:42:51,840 NO INTERVENTION, JUST SICKLE 3611 02:42:51,840 --> 02:42:53,480 CELL DISEASE. 3612 02:42:53,480 --> 02:43:00,800 AND THEN I'M SKIPPING THROUGH 3613 02:43:00,800 --> 02:43:06,600 THINGS, ALSO LOOKED AT PATHWAYS 3614 02:43:06,600 --> 02:43:09,160 UPREGULATING CATHEPSIN 3615 02:43:09,160 --> 02:43:12,160 REGULATION. 3616 02:43:12,160 --> 02:43:14,280 SO JNK, WE BEGAN TO INJECT MICE 3617 02:43:14,280 --> 02:43:21,440 FROM ONE TO THREE MONTHS WITH 3618 02:43:21,440 --> 02:43:23,440 JNK INHIBITOR TO STOP EXPRESSION 3619 02:43:23,440 --> 02:43:24,920 AND STOP ARTERIAL REMODEL. 3620 02:43:24,920 --> 02:43:26,920 THIS IS AN EXCITING RESULT, 3621 02:43:26,920 --> 02:43:33,560 SEVERAL YEARS AGO, THERE'S A 3622 02:43:33,560 --> 02:43:37,400 GENE DOSING EFFECT, SS, 3623 02:43:37,400 --> 02:43:38,600 IMMUNOHISTOCHEMISTRY, INCREASED 3624 02:43:38,600 --> 02:43:40,800 CATHEPSIN K IN CAROTID ARTERY 3625 02:43:40,800 --> 02:43:46,520 WALLS, WHEN THEY RECEIVED IP 3626 02:43:46,520 --> 02:43:51,200 INJECTIONS OF THE JNK INHIBITOR 3627 02:43:51,200 --> 02:43:53,720 THAT PRESERVED ARTERY STRUCTURE, 3628 02:43:53,720 --> 02:43:56,840 OUT TO FIVE MONTHS, INJECTED 3629 02:43:56,840 --> 02:43:58,400 DAILY, ISCHEMIC STROKES EARLY 3630 02:43:58,400 --> 02:44:00,920 ON, HEMORRHAGIC STROKES, BACK TO 3631 02:44:00,920 --> 02:44:02,240 ISCHEMIC, WE'RE SEEING THIS 3632 02:44:02,240 --> 02:44:03,880 INCREDIBLE REMODELING OF THE 3633 02:44:03,880 --> 02:44:06,520 SICKLE CELL ARTERIES BY FIVE 3634 02:44:06,520 --> 02:44:08,040 MONTHS THAT, AGAIN, JNK 3635 02:44:08,040 --> 02:44:11,440 INHIBITION WAS ABLE TO BLOCK 3636 02:44:11,440 --> 02:44:13,200 THAT REMODELING. 3637 02:44:13,200 --> 02:44:14,840 WE CARED ABOUT ELASTIC BREAKS 3638 02:44:14,840 --> 02:44:17,280 FROM THE AUTOPSIES IN CHILDREN 3639 02:44:17,280 --> 02:44:20,200 BUT THE QUESTION WAS WHAT IS 3640 02:44:20,200 --> 02:44:20,480 HAPPENING? 3641 02:44:20,480 --> 02:44:23,400 MY WONDERFUL STAFF SCIENTIST 3642 02:44:23,400 --> 02:44:27,880 HANNAH SUNG ACTUALLY BEGAN TO DO 3643 02:44:27,880 --> 02:44:30,480 NASCENT STAINING, BY FIVE MONTHS 3644 02:44:30,480 --> 02:44:31,600 COLLAGEN INDICATED BY BLUE 3645 02:44:31,600 --> 02:44:38,120 STAINING IS KIND OF GONE IN THAT 3646 02:44:38,120 --> 02:44:42,400 SICKLE CELL CAROTID ARTERY, 3647 02:44:42,400 --> 02:44:44,120 HOMOGENIZED TO QUANTIFY COLLAGEN 3648 02:44:44,120 --> 02:44:47,360 IN THE WALL, IF YOU GIVE THEM 3649 02:44:47,360 --> 02:44:48,600 THE JNK INHIBITOR YOU CAN 3650 02:44:48,600 --> 02:44:52,240 PROTECT COLLAGEN IN THE VESSEL 3651 02:44:52,240 --> 02:44:52,880 WALL. 3652 02:44:52,880 --> 02:44:55,960 SO THERE'S A PICTURE OF HER, 3653 02:44:55,960 --> 02:44:57,480 BECAUSE THERE'S SO MUCH 3654 02:44:57,480 --> 02:44:58,480 VARIABILITY, 11% OF CHILDREN 3655 02:44:58,480 --> 02:45:00,800 HAVING A MAJOR STROKE, A THIRD 3656 02:45:00,800 --> 02:45:04,040 SILENT STROKE, THAT WAS 3657 02:45:04,040 --> 02:45:05,040 REFLECTED IN FLUCTUATION AMONG 3658 02:45:05,040 --> 02:45:06,160 OUR MICE. 3659 02:45:06,160 --> 02:45:09,680 HANNAH FOUND THIS METHOD OF 3660 02:45:09,680 --> 02:45:11,080 USING LABEL-FREE MAGNETIC 3661 02:45:11,080 --> 02:45:16,000 RESONANCE ANGIOGRAM, WE WANTED 3662 02:45:16,000 --> 02:45:17,440 TO DO LONGITUDINAL SCANS, SCAN 3663 02:45:17,440 --> 02:45:19,160 OVER A PERIOD OF TIME. 3664 02:45:19,160 --> 02:45:21,160 SOMETIMES THEY DIE BUT WE WANTED 3665 02:45:21,160 --> 02:45:22,360 TO MINIMIZE VARIABILITY AND BE 3666 02:45:22,360 --> 02:45:24,960 ABLE TO TRACK IT FOR INDIVIDUAL 3667 02:45:24,960 --> 02:45:27,480 ANIMALS. 3668 02:45:27,480 --> 02:45:28,480 SO AGAIN THE BENEFITS, 3669 02:45:28,480 --> 02:45:30,560 NON-INVASIVE, GOOD SIGNAL 3670 02:45:30,560 --> 02:45:31,520 WITHOUT CONTRAST, BETTER WITH 3671 02:45:31,520 --> 02:45:38,960 CONTRAST, WE'RE HOPE TO THINKING 3672 02:45:38,960 --> 02:45:41,160 ABOUT IT. 3673 02:45:41,160 --> 02:45:43,120 I WOULD SAY I'VE BEEN DOING IT 3674 02:45:43,120 --> 02:45:44,760 FOR 13 YEARS, BUT SOME REVIEWS I 3675 02:45:44,760 --> 02:45:50,680 USED TO GET FROM MY WONDERFUL 3676 02:45:50,680 --> 02:45:53,080 NIH REVIEWERS WAS THAT MICE 3677 02:45:53,080 --> 02:45:56,800 NEVER SHOWN TO HAVE A STROKE, NO 3678 02:45:56,800 --> 02:45:58,640 ONE SHOWS IT HAVING A STROKE, 3679 02:45:58,640 --> 02:45:59,640 WHY SHOULD I BELIEVE YOU'RE 3680 02:45:59,640 --> 02:46:01,400 SEEING A STROKE IN THE MICE? 3681 02:46:01,400 --> 02:46:02,720 THAT'S NOT FAIR. 3682 02:46:02,720 --> 02:46:04,880 THAT'S WHETHER WE WENT WITH 3683 02:46:04,880 --> 02:46:07,080 LONGITUDINAL SCANS TO START 3684 02:46:07,080 --> 02:46:08,520 EXACERBATED ARTERIAL DAMAGE AND 3685 02:46:08,520 --> 02:46:11,920 SHOW PROGRESSING OVER TIME. 3686 02:46:11,920 --> 02:46:14,760 LOOKING AT TEN BRAINS THREE 3687 02:46:14,760 --> 02:46:15,960 MIGHT HAVE ABNORMALITIES IN 3688 02:46:15,960 --> 02:46:17,280 SCAN, BRAIN SCAN. 3689 02:46:17,280 --> 02:46:27,760 HOW CAN WE FOLLOW INDIVIDUAL 3690 02:46:28,200 --> 02:46:28,360 ANIMALS? 3691 02:46:28,360 --> 02:46:30,880 SAME MICE AT ONE AND THREE 3692 02:46:30,880 --> 02:46:31,120 MONTHS. 3693 02:46:31,120 --> 02:46:33,000 AS THEY GROW THE CROSS-SECTIONAL 3694 02:46:33,000 --> 02:46:34,960 AREA IS LARGER, BUT YOU SEE 3695 02:46:34,960 --> 02:46:45,480 VARIATION AMONG OUR SICKLE CELL 3696 02:46:45,720 --> 02:46:47,280 ANIMALS, WIDE VARIABILITY. 3697 02:46:47,280 --> 02:46:50,040 A NEW GRAD STUDENT HAS TAKEN 3698 02:46:50,040 --> 02:46:54,840 THIS ON, DOING SCANS OF OUR 3699 02:46:54,840 --> 02:46:56,560 SICKLE CELL MICE, AND WILL 3700 02:46:56,560 --> 02:46:58,680 EXPORT DATA AND RECREATE THE 3701 02:46:58,680 --> 02:47:00,560 VESSELS USING MIMICS. 3702 02:47:00,560 --> 02:47:02,520 WE'RE FOLLOWING THE SAME ANIMAL 3703 02:47:02,520 --> 02:47:05,120 ONE, THREE, FIVE MONTHS, SHE 3704 02:47:05,120 --> 02:47:07,640 FINISHED SEVEN MONTH SCANS AS 3705 02:47:07,640 --> 02:47:09,920 WELL, NOW QUANTIFYING FOR SINGLE 3706 02:47:09,920 --> 02:47:10,360 MOUSE. 3707 02:47:10,360 --> 02:47:13,840 I'M SHOWING AGGREGATED DATA 3708 02:47:13,840 --> 02:47:15,720 BECAUSE WE'RE STILL FINDING BEST 3709 02:47:15,720 --> 02:47:18,400 WAYS TO TRACK INDIVIDUAL MICE. 3710 02:47:18,400 --> 02:47:20,320 THIS IS N OF 18 WHERE WE'RE 3711 02:47:20,320 --> 02:47:25,920 SEEING THAT THERE'S AN INCREASE 3712 02:47:25,920 --> 02:47:28,200 IN THE CAROTID ARTERY SIZE IN 3713 02:47:28,200 --> 02:47:29,840 SICKLE CELL ANIMALS, MORE 3714 02:47:29,840 --> 02:47:31,720 SIGNIFICANT IN MALES THAN 3715 02:47:31,720 --> 02:47:33,680 FEMALES, EVEN AS WE SEE IN DATA 3716 02:47:33,680 --> 02:47:34,000 HERE. 3717 02:47:34,000 --> 02:47:35,560 WHAT WE'VE BEEN THINKING ABOUT 3718 02:47:35,560 --> 02:47:40,440 IS THE ONLY FUNCTIONAL CURE FOR 3719 02:47:40,440 --> 02:47:45,800 SICKLE CELL IS BONE MARROW 3720 02:47:45,800 --> 02:47:48,000 TRANSPLANT OR HEMATOPOIETIC STEM 3721 02:47:48,000 --> 02:47:50,080 CELL TRANSGENIC PLANT, STILL 3722 02:47:50,080 --> 02:47:51,280 WITH SOME COMPLICATION OF 3723 02:47:51,280 --> 02:47:52,280 CEREBRAL INFARCTION, WONDERING 3724 02:47:52,280 --> 02:47:53,360 IF WE COULD SEE THAT IN THE 3725 02:47:53,360 --> 02:47:55,000 MOUSE MODEL EVEN AFTER 3726 02:47:55,000 --> 02:47:56,640 CORRECTING FOR THE GENE MUTATION 3727 02:47:56,640 --> 02:47:59,720 BY REPLACING THE CELLS WITH 3728 02:47:59,720 --> 02:48:03,200 HEALTHY CELLS COULD WE STILL OR 3729 02:48:03,200 --> 02:48:05,080 TRACK DAMAGE TO ARTERY WALL, 3730 02:48:05,080 --> 02:48:07,040 GREAT WORK AT NIH CLINICAL 3731 02:48:07,040 --> 02:48:08,360 TRIALS FOR SICKLE CELL DISEASE, 3732 02:48:08,360 --> 02:48:15,680 HOPING TO GET INVOLVED WITH THAT 3733 02:48:15,680 --> 02:48:16,920 WORK. 3734 02:48:16,920 --> 02:48:20,720 HYPOTHESIS IF WE DO EARLY SCD 3735 02:48:20,720 --> 02:48:25,520 WILL THAT PROTECT SS ARTERIES 3736 02:48:25,520 --> 02:48:27,600 FROM ARTERIAL MODELING? 3737 02:48:27,600 --> 02:48:30,480 WE COMPARED TWO MONTHS AND FOUR 3738 02:48:30,480 --> 02:48:32,000 MONTHS, ARTERIAL DIAMETERS, 3739 02:48:32,000 --> 02:48:33,720 SEEING THE EXPANSIVE REMODELING. 3740 02:48:33,720 --> 02:48:44,320 AND WE SWITCHED , HANNAH SWITCHD 3741 02:48:47,800 --> 02:48:51,560 TO BUSULFAN, TO MONITOR THE 3742 02:48:51,560 --> 02:48:52,920 BLOOD VESSELS. 3743 02:48:52,920 --> 02:48:56,240 AND WOULD CONFIRM HSCT WAS 3744 02:48:56,240 --> 02:48:58,560 SUCCESSFUL RUNNING PCR, THE TAIL 3745 02:48:58,560 --> 02:49:02,400 SHOULD STILL CARRY THE SICKLE 3746 02:49:02,400 --> 02:49:03,360 CELL MUTATION, COLLECT 3747 02:49:03,360 --> 02:49:04,040 PERIPHERAL BLOOD FROM 3748 02:49:04,040 --> 02:49:05,120 HEMATOPOIETIC STEM CELLS AND 3749 02:49:05,120 --> 02:49:10,360 MAKE SURE THEY WERE AT LEAST 3750 02:49:10,360 --> 02:49:12,680 HETEROZYGOUS FOR WILD TYPE 3751 02:49:12,680 --> 02:49:13,000 ALLELE. 3752 02:49:13,000 --> 02:49:15,280 AFTER HSCT IT WAS QUITING, DATA 3753 02:49:15,280 --> 02:49:18,280 SHE JUST FINISHED COLLECTING, 3754 02:49:18,280 --> 02:49:20,680 AND THATs EARLY HSCT WERE ABLE 3755 02:49:20,680 --> 02:49:24,600 TO PROTECT ARTERIAL DIAMETER OF 3756 02:49:24,600 --> 02:49:27,400 BLOOD VESSELS, AT AGE 5 MONTHS, 3757 02:49:27,400 --> 02:49:29,760 SO THREE MONTHS AFTER. 3758 02:49:29,760 --> 02:49:32,480 IN THE SHAM MICE THEY STILL HAVE 3759 02:49:32,480 --> 02:49:34,880 SIGNIFICANT DIFFERENCE. 3760 02:49:34,880 --> 02:49:36,240 IN LATE MICE, HSCT AT FOUR 3761 02:49:36,240 --> 02:49:37,880 MONTHS, WE'RE STILL SEEING A 3762 02:49:37,880 --> 02:49:40,920 SIGNIFICANT DIFFERENCE BETWEEN 3763 02:49:40,920 --> 02:49:42,680 THE CONTROLS AND THE S S MICE. 3764 02:49:42,680 --> 02:49:43,560 WHAT DOES THAT SAY? 3765 02:49:43,560 --> 02:49:48,040 THE EARLIER YOU DO THE HSCT, YOU 3766 02:49:48,040 --> 02:49:51,200 PROTECT THE VESSELS MORE SO. 3767 02:49:51,200 --> 02:49:52,640 WE'LL KEEP FOLLOWING THESE AND 3768 02:49:52,640 --> 02:49:58,440 LOOK AT EXTRA TREATMENT OF JNK 3769 02:49:58,440 --> 02:50:03,480 IF YOU DO THE HSCT LATER. 3770 02:50:03,480 --> 02:50:09,600 WHAT IS GREAT ABOUT THE MAGNETIC 3771 02:50:09,600 --> 02:50:10,440 RESONANCE ANGIOGRAPHY LOOKING AT 3772 02:50:10,440 --> 02:50:13,440 FEATURE WHERE IS WE MIGHT SEE 3773 02:50:13,440 --> 02:50:17,280 STENOSIS OR BLOCKAGE, IN OTHER 3774 02:50:17,280 --> 02:50:18,320 STUDIES PEOPLE MANIPULATE LASERS 3775 02:50:18,320 --> 02:50:19,920 TO CAUSE A STROKE BUT THAT'S NOT 3776 02:50:19,920 --> 02:50:23,360 WHAT YOU SEE IN PEOPLE LIVING 3777 02:50:23,360 --> 02:50:25,040 WITH SICKLE CELL. 3778 02:50:25,040 --> 02:50:31,280 WE DIDN'T WANT BLOWOUT STROKES. 3779 02:50:31,280 --> 02:50:38,840 KNOW SHE CAN LOOK AT TORTUOSITY. 3780 02:50:38,840 --> 02:50:40,280 ARE THEY REALLY HAVING STROKES? 3781 02:50:40,280 --> 02:50:46,600 NOW WITH M.R. SHE CAN LOOK FOR 3782 02:50:46,600 --> 02:50:48,280 OCCLUDED VESSELS, THAT NOW COULD 3783 02:50:48,280 --> 02:50:50,680 GIVE A STRONG INDICATION THAT 3784 02:50:50,680 --> 02:50:52,120 ACTUALLY MAYBE A STROKE THESE 3785 02:50:52,120 --> 02:50:56,680 MICE ARE HAVING TO CONVINCE THE 3786 02:50:56,680 --> 02:50:57,880 COMMUNITY, TRADITIONAL BRAIN 3787 02:50:57,880 --> 02:50:58,760 SCANS SHOW ABNORMALITIES ON THE 3788 02:50:58,760 --> 02:51:02,600 BRAINS, NOT IN ALL THE MICE, AT 3789 02:51:02,600 --> 02:51:04,680 LEAST 10 TO 15%. 3790 02:51:04,680 --> 02:51:06,440 AND ONE MORE THING, WHAT IS THE 3791 02:51:06,440 --> 02:51:09,040 CHALLENGE HERE, WE SAID WE WERE 3792 02:51:09,040 --> 02:51:10,360 MEASURING ARTERIAL DIAMETER BUT 3793 02:51:10,360 --> 02:51:14,520 AS YOU GO THE LENGTH OF THAT 3794 02:51:14,520 --> 02:51:15,840 ARTERY, CHANGES FLUCTUATE, AND 3795 02:51:15,840 --> 02:51:17,160 EVEN THIS ONE MOUSE 3796 02:51:17,160 --> 02:51:19,320 INTERESTINGLY FROM THREE MONTHS 3797 02:51:19,320 --> 02:51:21,320 TO FIVE MONTHS SHE BEGAN TO 3798 02:51:21,320 --> 02:51:24,040 NOTICE WHEN WE LOOKED AT THE 3799 02:51:24,040 --> 02:51:25,440 PERCENT AREA CHANGE, OR FROM 3 3800 02:51:25,440 --> 02:51:29,320 TO 5 MONTHS, SOME MICE HAD A 3801 02:51:29,320 --> 02:51:30,760 REDUCTION IN THE AREA WHICH 3802 02:51:30,760 --> 02:51:32,560 DOESN'T MAKE SENSE IF THE MOUSE 3803 02:51:32,560 --> 02:51:35,880 IS GROWING OR AT LEAST SHOULD 3804 02:51:35,880 --> 02:51:36,160 MAINTAIN. 3805 02:51:36,160 --> 02:51:37,600 WE'VE BEEN TRYING TO CHANGE THE 3806 02:51:37,600 --> 02:51:41,120 WAIT WE ANALYZE ALONG THE ENTIRE 3807 02:51:41,120 --> 02:51:47,360 LENGTH, LOOKING AT VOLUME, NOW 3808 02:51:47,360 --> 02:51:48,760 TRYING TO IDENTIFY STENOSES SO 3809 02:51:48,760 --> 02:51:51,520 WE CAN SACRIFICE WHEN WE SEE THE 3810 02:51:51,520 --> 02:51:54,480 STENOSIS AND UNDERSTAND WHAT'S 3811 02:51:54,480 --> 02:51:54,920 HAPPENING. 3812 02:51:54,920 --> 02:51:57,400 IT INDUCES SEX AND AGE-RELATED 3813 02:51:57,400 --> 02:51:59,960 DIFFERENCE IN LARGE ARTERY 3814 02:51:59,960 --> 02:52:02,560 REMODEL AND SS MALES SHOW 3815 02:52:02,560 --> 02:52:03,400 GREATEST EXPANSION CORRELATING 3816 02:52:03,400 --> 02:52:05,960 WITH HUMANS IN THE SEX-SPECIFIC 3817 02:52:05,960 --> 02:52:08,360 DIFFERENCES IN LIFE EXPECTANCY. 3818 02:52:08,360 --> 02:52:09,160 CURATIVE HEMATOPOIETIC STEM CELL 3819 02:52:09,160 --> 02:52:11,320 TRANSPLANT DOES PROTECT THESE 3820 02:52:11,320 --> 02:52:12,520 ARTERIES FROM PATHOLOGICAL 3821 02:52:12,520 --> 02:52:13,840 REMODEL, ESPECIALLY AT EARLIER 3822 02:52:13,840 --> 02:52:14,160 AGE. 3823 02:52:14,160 --> 02:52:15,680 AND AGAIN THROUGH THIS 3824 02:52:15,680 --> 02:52:16,880 LONGITUDINAL MRA WE'RE ABLE TO 3825 02:52:16,880 --> 02:52:20,480 GET MORE OF A TIME LINE FOR THIS 3826 02:52:20,480 --> 02:52:23,920 DAMAGE AND NOW WE'LL LOOK AT 3827 02:52:23,920 --> 02:52:26,840 HISTOLOGICAL ANALYSES AND MATCH 3828 02:52:26,840 --> 02:52:27,960 WITH SUGGESTED THERAPEUTICS. 3829 02:52:27,960 --> 02:52:33,080 ALSO TO GET TO THE CASE MADE AND 3830 02:52:33,080 --> 02:52:38,000 HAVE DATA FOR CATHEPSIN KNOCKOUT 3831 02:52:38,000 --> 02:52:39,560 MICE, MTA HAS FINALLY MOVED 3832 02:52:39,560 --> 02:52:46,440 THROUGH, SO THE MICE CAN BE SENT 3833 02:52:46,440 --> 02:52:47,080 HERE. 3834 02:52:47,080 --> 02:52:50,680 NO MATTER WHAT YOU'RE STUDYING, 3835 02:52:50,680 --> 02:52:55,280 IT APPLIES TO SICKLE CELL 3836 02:52:55,280 --> 02:52:56,120 DISEASE. 3837 02:52:56,120 --> 02:52:57,720 INTERESTINGLY ENGINEERING 3838 02:52:57,720 --> 02:52:59,920 QUESTIONS, INTERESTING CHEMICAL 3839 02:52:59,920 --> 02:53:06,040 QUESTIONS AND NEW METHODOLOGIES. 3840 02:53:06,040 --> 02:53:07,680 THANK YOU FOR LISTENING. 3841 02:53:07,680 --> 02:53:08,960 I THANK NIBIB FOR FUNDING TO 3842 02:53:08,960 --> 02:53:11,080 CONTINUE THIS WORK AND THIS WAS 3843 02:53:11,080 --> 02:53:16,560 MY RESEARCH GROUP IN SEPTEMBER 3844 02:53:16,560 --> 02:53:21,600 AT THE SICKLE CELL WALK-RUN IN 3845 02:53:21,600 --> 02:53:22,000 GEORGIA. 3846 02:53:22,000 --> 02:53:23,000 THERE'S LEANNA WITH WONDERFUL 3847 02:53:23,000 --> 02:53:26,840 KIDS, HER HUSBAND RAN OFF 3848 02:53:26,840 --> 02:53:27,720 SOMEWHERE. 3849 02:53:27,720 --> 02:53:34,160 THERE WE ARE. 3850 02:53:34,160 --> 02:53:36,240 THANK YOU VERY MUCH. 3851 02:53:36,240 --> 02:53:36,520 [APPLAUSE] 3852 02:53:36,520 --> 02:53:43,160 >> WHAT DOES THE RETINAL 3853 02:53:43,160 --> 02:53:45,560 VASCULATURE LOOK LIKE? 3854 02:53:45,560 --> 02:53:47,000 >> DEFINITELY THERE'S DAMAGE, 3855 02:53:47,000 --> 02:53:49,800 YOU CAN SEE IN EARLY 20s, LATE 3856 02:53:49,800 --> 02:53:51,320 TEENS, BUT EVEN BEYOND THAT 3857 02:53:51,320 --> 02:53:53,960 SICKLE TRAIT WHICH PEOPLE ARE 3858 02:53:53,960 --> 02:53:56,040 TOLD IS BENIGN THERE'S RETINAL 3859 02:53:56,040 --> 02:53:57,800 DAMAGE, ONE OF THE COMPLICATIONS 3860 02:53:57,800 --> 02:53:58,800 OF SICKLE CELL TRAIT. 3861 02:53:58,800 --> 02:54:01,640 >> YOU COULD DO A LOT MORE WITH 3862 02:54:01,640 --> 02:54:03,240 THE RETINAL VESSELS, CEREBRAL 3863 02:54:03,240 --> 02:54:05,480 VESSELS ARE HARD TO GET TO I 3864 02:54:05,480 --> 02:54:06,480 GUESS AND HARDER TO IMAGE. 3865 02:54:06,480 --> 02:54:07,440 >> I WAS INTERESTED IN TALKING 3866 02:54:07,440 --> 02:54:09,720 WITH YOU ABOUT THAT. 3867 02:54:09,720 --> 02:54:15,760 REMEMBER THE A.S. IS A LARGER 3868 02:54:15,760 --> 02:54:17,960 POPULATION BUT THEY ARE ALL HERE 3869 02:54:17,960 --> 02:54:18,680 AT THE CLINICAL CENTER. 3870 02:54:18,680 --> 02:54:22,560 >> YOU'RE GOING TO GUESS THE 3871 02:54:22,560 --> 02:54:23,320 THEME OF MY QUESTION. 3872 02:54:23,320 --> 02:54:25,960 HAVE YOU LOOKED AT THE DIFFERENT 3873 02:54:25,960 --> 02:54:28,440 CELLS THAT ARE PARTICIPATING IN 3874 02:54:28,440 --> 02:54:30,640 THAT REMODELING WITHIN THE 3875 02:54:30,640 --> 02:54:33,400 VASCULATURE BECAUSE, YOU KNOW, 3876 02:54:33,400 --> 02:54:35,680 YOUNG EVEN IN UTERO VERSUS BIRTH 3877 02:54:35,680 --> 02:54:41,080 MICE WILL HAVE A DIFFERENT 3878 02:54:41,080 --> 02:54:44,320 IMMUNE RESPONSE TO TISSUE 3879 02:54:44,320 --> 02:54:45,960 DAMAGE, CATHEPSINS COME FROM 3880 02:54:45,960 --> 02:54:49,320 MYELOID CELLS, CURIOUS. 3881 02:54:49,320 --> 02:54:51,560 >> I DIDN'T SHOW THE HUMAN 3882 02:54:51,560 --> 02:54:51,760 WORK. 3883 02:54:51,760 --> 02:54:53,960 WE STARTED WITH GETTING BLOOD 3884 02:54:53,960 --> 02:54:59,640 FROM PEOPLE WITH SICKLE CELL, 3885 02:54:59,640 --> 02:55:01,640 USING WITH ENDOTHELIA CELLS IN 3886 02:55:01,640 --> 02:55:08,480 THE SHEER STRESS FLOW CHAMBER. 3887 02:55:08,480 --> 02:55:10,480 THERE'S CHRONIC AND SYSTEMIC 3888 02:55:10,480 --> 02:55:13,760 INFLAMMATION, HIGH LEVELS OF 3889 02:55:13,760 --> 02:55:16,080 CIRCULATING TNF ALPHA. 3890 02:55:16,080 --> 02:55:21,320 IF YOU TAKE PBMCs COMPARED TO 3891 02:55:21,320 --> 02:55:23,480 A.A., ADHERE BETTER, EVEN 3892 02:55:23,480 --> 02:55:24,280 WITHOUT PRE-TREATMENT WITH TNF 3893 02:55:24,280 --> 02:55:28,080 ALPHA AND TURN ON EXPRESSION OF 3894 02:55:28,080 --> 02:55:30,240 CYSTEINE CATHEPSINS, IT'S A 3895 02:55:30,240 --> 02:55:32,120 POSITIVE FEEDBACK, MORE 3896 02:55:32,120 --> 02:55:37,920 MONOCYTES, MOR ADHESIVE, THEY 3897 02:55:37,920 --> 02:55:39,160 AIN'T GOT NO CHANCE. 3898 02:55:39,160 --> 02:55:43,080 IN OTHER STUDIES WE BEGAN TO 3899 02:55:43,080 --> 02:55:46,160 LOOK AT INFLAMMATORY MONOCYTES 3900 02:55:46,160 --> 02:55:46,960 VERSUS ANTI-ININFLAMMATORY, 3901 02:55:46,960 --> 02:55:49,680 WE'VE SEEN THAT IN HUMANS AND 3902 02:55:49,680 --> 02:55:56,360 MICE, IT'S THE DIFFERENT MARKER, 3903 02:55:56,360 --> 02:55:59,440 CD16, THERE'S A GREATER NUMBER 3904 02:55:59,440 --> 02:56:01,520 OF INFLAMMATORY MONOCYTES IN 3905 02:56:01,520 --> 02:56:03,240 INDIVIDUALS WITH SS, EVEN 3906 02:56:03,240 --> 02:56:06,760 MONOCYTES ARE MORE 3907 02:56:06,760 --> 02:56:08,760 PROTEOLYTICALLY ACTIVE BEFORE 3908 02:56:08,760 --> 02:56:11,840 THEY BIND AND STICK TO CELLS. 3909 02:56:11,840 --> 02:56:14,080 HANNAH CARES ABOUT T CELLS, I 3910 02:56:14,080 --> 02:56:15,880 TELL HER TO GET THAT STUFF OUT 3911 02:56:15,880 --> 02:56:17,480 OF MY FACE BUT NOW WE'RE HERE 3912 02:56:17,480 --> 02:56:18,680 AND WE SHOULD TALK. 3913 02:56:18,680 --> 02:56:20,920 IF THERE'S OTHER DATA WE HAVE 3914 02:56:20,920 --> 02:56:23,840 ALREADY COLLECTED WE'D LOVE TO 3915 02:56:23,840 --> 02:56:25,480 SHARE AND SHOW INTERESTING 3916 02:56:25,480 --> 02:56:26,600 THINGS TO LOOK AT. 3917 02:56:26,600 --> 02:56:27,560 >> GREAT TALK. 3918 02:56:27,560 --> 02:56:30,520 >> ALL RIGHT, THANK YOU. 3919 02:56:30,520 --> 02:56:35,760 3920 02:56:35,760 --> 02:56:38,200 3921 02:56:38,200 --> 02:56:42,720 >> THIS IS NOT AN IMMUNOLOGY 3922 02:56:42,720 --> 02:56:43,000 QUESTION. 3923 02:56:43,000 --> 02:56:46,160 AM I CORRECT IT'S THE TURBULENT 3924 02:56:46,160 --> 02:56:48,320 FLOW AFTER THE STENOSIS CAUSING 3925 02:56:48,320 --> 02:56:51,480 THE DAMAGE AND SUBSEQUENT 3926 02:56:51,480 --> 02:56:51,800 REMODELING? 3927 02:56:51,800 --> 02:56:54,800 >>WE DON'T KNOW IF IT GOES 3928 02:56:54,800 --> 02:56:56,640 TURBULENT BUT DEFINITELY 3929 02:56:56,640 --> 02:57:05,960 DISTURBED FLOW, ENDOTHELIAL 3930 02:57:05,960 --> 02:57:06,160 CELLS. 3931 02:57:06,160 --> 02:57:10,000 >> CAN YOU ADD SOMETHING TO THE 3932 02:57:10,000 --> 02:57:14,160 CIRCULATION TO LAMINARIZE THE 3933 02:57:14,160 --> 02:57:17,400 FLOW, LIKE DRUG RESISTANT 3934 02:57:17,400 --> 02:57:17,640 POLYMERS. 3935 02:57:17,640 --> 02:57:20,720 >> OH, INTERESTING. 3936 02:57:20,720 --> 02:57:24,200 THE PROBLEM THAT -- MOST PEOPLE 3937 02:57:24,200 --> 02:57:29,320 CALL THIS SICKLE CELL ANEMIA, 3938 02:57:29,320 --> 02:57:35,840 HEMATOCRIT IS 45% WITHOUT, IN 3939 02:57:35,840 --> 02:57:37,040 SICKLE CELL IT'S 25%. 3940 02:57:37,040 --> 02:57:39,440 IN THE PROCESS OF THEM BREAKING 3941 02:57:39,440 --> 02:57:42,080 UP THE MEMBRANES EACH INDIVIDUAL 3942 02:57:42,080 --> 02:57:45,800 RED CELL MEMBRANE GETS FOR 3943 02:57:45,800 --> 02:57:49,000 VISCOUS, THAN HAVING THEM IN 3944 02:57:49,000 --> 02:57:51,600 AGGREGATE DOES NOT GET MORE 3945 02:57:51,600 --> 02:57:53,000 VISCOUS BECAUSE THERE'S LESS RED 3946 02:57:53,000 --> 02:57:55,680 BLOOD CELLS SO WHEN YOU ASK, 3947 02:57:55,680 --> 02:57:58,840 THERE'S STILL INDIVIDUAL RED 3948 02:57:58,840 --> 02:57:59,920 CELLS CREATING THEIR OWN 3949 02:57:59,920 --> 02:58:00,320 CHANGES. 3950 02:58:00,320 --> 02:58:02,640 YOU DON'T WANT TO THIN OUT RED 3951 02:58:02,640 --> 02:58:05,280 CELLS BECAUSE YOU'RE ALREADY AT 3952 02:58:05,280 --> 02:58:07,560 LIMITS HOW YOU TRANSPORT OXYGEN. 3953 02:58:07,560 --> 02:58:09,080 THERE HAVE BEEN OTHER DRUGS THAT 3954 02:58:09,080 --> 02:58:11,040 PEOPLE HAVE TRIED IN THE PAST 3955 02:58:11,040 --> 02:58:16,880 WHERE THIS HAS BEEN A PROBLEM, 3956 02:58:16,880 --> 02:58:18,200 POLYMERS WOULD PREVENT 3957 02:58:18,200 --> 02:58:23,000 HEMOGLOBIN MOLECULES FROM 3958 02:58:23,000 --> 02:58:24,920 POLYMERIZING, BUT EXCRETING 3959 02:58:24,920 --> 02:58:26,680 THROUGH THE KIDNEYS, YOU GOT TO 3960 02:58:26,680 --> 02:58:29,800 KEEP ENOUGH MOLECULES IN THE 3961 02:58:29,800 --> 02:58:30,880 BLOODSTREAM WITH THE HEMOGLOBIN 3962 02:58:30,880 --> 02:58:34,160 TO STOP THAT. 3963 02:58:34,160 --> 02:58:34,560 BUMMER. 3964 02:58:34,560 --> 02:58:36,760 THE ONE TREATMENT THAT DOES 3965 02:58:36,760 --> 02:58:39,080 WORK, WHERE CLINICAL TRIALS ARE 3966 02:58:39,080 --> 02:58:40,960 GOING NOW, ONE DRUG ON THE 3967 02:58:40,960 --> 02:58:41,720 MARKET THAT'S REALLY EFFECTIVE, 3968 02:58:41,720 --> 02:58:44,240 THERE ARE THREE BUT ONE REALLY 3969 02:58:44,240 --> 02:58:50,440 EFFECTIVE, TURNS BACK ON FETAL 3970 02:58:50,440 --> 02:58:58,000 HEMOGLOBIN, IN THE RED CELL 3971 02:58:58,000 --> 02:58:59,240 INTERFERES WITH SICKLE 3972 02:58:59,240 --> 02:59:00,680 POLYMERIZATION, VISCOSITY GOES 3973 02:59:00,680 --> 02:59:00,880 DOWN. 3974 02:59:00,880 --> 02:59:01,760 THAT'S WHY PEOPLE TARGET 3975 02:59:01,760 --> 02:59:03,160 PROTECTING THE RED CELL BUT I 3976 02:59:03,160 --> 02:59:04,080 HAVEN'T HEARD PEOPLE TALK ABOUT 3977 02:59:04,080 --> 02:59:09,200 HOW DO YOU MAKE THE FLOW MORE -- 3978 02:59:09,200 --> 02:59:13,800 LESS VISCOUS, BECAUSE RED CELLS 3979 02:59:13,800 --> 02:59:16,120 ARE INCREASING VISCOSITY. 3980 02:59:16,120 --> 02:59:22,560 LONG ANSWER, SORRY. 3981 02:59:22,560 --> 02:59:23,080 >> THANK YOU. 3982 02:59:23,080 --> 02:59:23,360 >> OKAY. 3983 02:59:23,360 --> 02:59:24,200 >> YOU SAID PORT OF THE PROBLEM 3984 02:59:24,200 --> 02:59:26,160 WITH CLOTS AND THINGS FORMING 3985 02:59:26,160 --> 02:59:27,840 ARE CELLS ARE STICKING TO THE 3986 02:59:27,840 --> 02:59:29,560 BLOOD VESSEL WELL. 3987 02:59:29,560 --> 02:59:32,400 HAVE PEOPLE ATTACKED THAT FROM A 3988 02:59:32,400 --> 02:59:33,080 PHARMACOLOGICAL VIEWPOINT? 3989 02:59:33,080 --> 02:59:34,280 >> YEAH, THERE'S A DRUG OUT 3990 02:59:34,280 --> 02:59:43,960 NOW, I THINK IT'S BEING TESTED, 3991 02:59:43,960 --> 02:59:45,120 KRIZUNLAZIMAB, P SELECTIN 3992 02:59:45,120 --> 02:59:48,920 ANTIBODY, IT HAS, LAST PAPER I 3993 02:59:48,920 --> 02:59:52,920 READ, WAS EFFECTIVE IN REDUCING 3994 02:59:52,920 --> 03:00:00,400 NUMBER OF VASOOCCLUSIONS OR 3995 03:00:00,400 --> 03:00:00,840 VASOOCCLUSIVE CRISES. 3996 03:00:00,840 --> 03:00:01,240 >> THANK YOU. 3997 03:00:01,240 --> 03:00:03,720 >> ALL RIGHT. 3998 03:00:03,720 --> 03:00:05,800 THANK YOU VERY MUCH. 3999 03:00:05,800 --> 03:00:08,760 [APPLAUSE] 4000 03:00:08,760 --> 03:00:11,840 >> WE'LL HAVE FOUR SHORT 4001 03:00:11,840 --> 03:00:20,520 TEN-MINUTE TALKS. 4002 03:00:20,520 --> 03:00:23,240 I WANT TO ANNOUNCE TOMORROW 4003 03:00:23,240 --> 03:00:33,320 WE'RE GOING TO HAVE A LUNCH WITH 4004 03:00:33,320 --> 03:00:34,200 DR. GEORGAKOUDI FOR WALS, IF YOU 4005 03:00:34,200 --> 03:00:36,240 WOULD LIKE TO JOIN US FOR DINNER 4006 03:00:36,240 --> 03:00:41,760 LET US KNOW. 4007 03:00:41,760 --> 03:00:43,160 CONTACT STEVE. 4008 03:00:43,160 --> 03:00:46,320 LET US KNOW IF THERE'S ANY 4009 03:00:46,320 --> 03:00:47,080 DIETARY RESTRICTION. 4010 03:00:47,080 --> 03:00:49,800 THE LUNCH WILL BE FROM TWELVE TO 4011 03:00:49,800 --> 03:00:51,280 ONE P.M. TOMORROW. 4012 03:00:51,280 --> 03:00:52,120 LET US KNOW. 4013 03:00:52,120 --> 03:00:54,640 ALL RIGHT. 4014 03:00:54,640 --> 03:00:59,880 NEXT SPEAKER IS DR. LICHEN 4015 03:00:59,880 --> 03:01:01,840 XIANG, NINR, WE WOULD LIKE TO 4016 03:01:01,840 --> 03:01:10,480 WELCOME HIM TO GIVE HIS TALK. 4017 03:01:10,480 --> 03:01:10,760 THANK YOU. 4018 03:01:10,760 --> 03:01:15,000 >>I'M A RESEARCH FELLOW, 4019 03:01:15,000 --> 03:01:23,720 INSTITUTE OF NURSING RESEARCH, 4020 03:01:23,720 --> 03:01:28,240 DIVISION OF NURSING, MY TOMMIC 4021 03:01:28,240 --> 03:01:29,120 IS RAPID POINT-OF-CARE TEST, 4022 03:01:29,120 --> 03:01:32,400 I'VE BEEN DOING THIS FROM MY 4023 03:01:32,400 --> 03:01:34,800 GRADUATE YEARS TO NOW. 4024 03:01:34,800 --> 03:01:37,520 THIS IS THE FIRST TEN-MINUTE 4025 03:01:37,520 --> 03:01:41,920 TALK FOR TODAY, I'LL TRY TO MAE 4026 03:01:41,920 --> 03:01:45,760 IT ON TIME. 4027 03:01:45,760 --> 03:01:49,360 YOU ARE MISSION TO SAVE PROBLEMS 4028 03:01:49,360 --> 03:01:52,000 IN THIRD WORLD COUNTRY WHICH 4029 03:01:52,000 --> 03:01:54,880 THERE ARE MANY CHILDRENS DIE 4030 03:01:54,880 --> 03:01:56,520 LIKE OVER 500,000 CHILDREN DIE 4031 03:01:56,520 --> 03:01:59,000 EVERY YEAR BECAUSE OF THE 4032 03:01:59,000 --> 03:02:01,960 DIARRHEA, ACCORDING TO THE 4033 03:02:01,960 --> 03:02:04,480 W.H.O., EVEN NOW TODAY. 4034 03:02:04,480 --> 03:02:07,320 EVEN TODAY. 4035 03:02:07,320 --> 03:02:10,600 SO, THERE ARE MANY KIDS DIE 4036 03:02:10,600 --> 03:02:12,240 BECAUSE OF LACK OF RESOURCES TO 4037 03:02:12,240 --> 03:02:16,280 TREAT IT OR DIAGNOSE IT. 4038 03:02:16,280 --> 03:02:20,440 THE CURRENT METHOD FOR TESTING 4039 03:02:20,440 --> 03:02:24,480 THE DIARRHEA INFECTIONS ARE 4040 03:02:24,480 --> 03:02:27,480 EITHER BEING PCR BASED OR CELL 4041 03:02:27,480 --> 03:02:30,680 CULTURE BASED, WASTING TIME, 4042 03:02:30,680 --> 03:02:33,240 SLOW, AND REQUIRED HIGHLY 4043 03:02:33,240 --> 03:02:35,440 SKILLED TECHNICIANS, THEY DEMAND 4044 03:02:35,440 --> 03:02:37,840 DEDICATED LABORATORY RESOURCES, 4045 03:02:37,840 --> 03:02:40,560 SO THOSE ARE NOT VERY APPLICABE 4046 03:02:40,560 --> 03:02:43,960 IN THE FIELD OR POOR COUNTRIES. 4047 03:02:43,960 --> 03:02:47,440 AND SO THERE'S A NEED FOR RAPID 4048 03:02:47,440 --> 03:02:48,800 REALTIME LOW COST POINT OF CARE 4049 03:02:48,800 --> 03:02:52,520 THAT CAN DIAGNOSE THE PATHOGENS 4050 03:02:52,520 --> 03:02:55,360 THAT ARE CAUSING DIARRHEA, AND 4051 03:02:55,360 --> 03:03:01,040 ALSO IN THE REAL LOW COST 4052 03:03:01,040 --> 03:03:01,280 SETTINGS. 4053 03:03:01,280 --> 03:03:04,880 SO, AT THE VERY BEGINNING OF 4054 03:03:04,880 --> 03:03:06,280 THIS RESEARCH WE FACED BIG 4055 03:03:06,280 --> 03:03:15,360 CHALLENGES BECAUSE IF WE'RE 4056 03:03:15,360 --> 03:03:17,760 DEALING WITH STEW THERE'S A LOT 4057 03:03:17,760 --> 03:03:20,600 OF INTERFERENCE THAT CAN 4058 03:03:20,600 --> 03:03:21,840 INTERFERE YOUR DNA APPLICATIONS, 4059 03:03:21,840 --> 03:03:23,680 ALL SUCH THINGS. 4060 03:03:23,680 --> 03:03:32,800 THAT'S WHY MOST OF THE LABS 4061 03:03:32,800 --> 03:03:36,880 USING THE LIKE SAMPLE PREP KIDS 4062 03:03:36,880 --> 03:03:43,440 TO ISOLATE DNA FROM THE STOOL OR 4063 03:03:43,440 --> 03:03:45,600 USING THE CENTRIFUGE AT LEAST, 4064 03:03:45,600 --> 03:03:45,920 RIGHT? 4065 03:03:45,920 --> 03:03:48,360 SO THE VERY BEGINNING WE WANTED 4066 03:03:48,360 --> 03:03:50,280 TO TRY TO HAVE SOMETHING THAT IS 4067 03:03:50,280 --> 03:03:53,320 LOW COST AND DON'T NEED A LAB TO 4068 03:03:53,320 --> 03:03:55,000 DO BIOSEPARATION. 4069 03:03:55,000 --> 03:04:05,560 THIS IS THE METHOD WE DEVELOPED, 4070 03:04:08,760 --> 03:04:10,080 MICRO SPHERE BIO-SEPARATION 4071 03:04:10,080 --> 03:04:12,000 METHOD, IT HAS A REALLY, REALLY 4072 03:04:12,000 --> 03:04:16,480 LOW DENSITY, SO IF WE PUT THAT 4073 03:04:16,480 --> 03:04:18,640 INTO A LIQUID IT WILL FLOAT TO 4074 03:04:18,640 --> 03:04:21,360 THE TOP VERY FAST. 4075 03:04:21,360 --> 03:04:22,560 SO HERE'S THE REALTIME MEDIA 4076 03:04:22,560 --> 03:04:28,040 CLIP THAT WE JUST MIX IT UP WITH 4077 03:04:28,040 --> 03:04:31,120 THESE AND WATER, PBS, YES, AND 4078 03:04:31,120 --> 03:04:32,560 THEN JUST ESSENTIALLY FLOAT TO 4079 03:04:32,560 --> 03:04:36,160 THE TOP, RIGHT? 4080 03:04:36,160 --> 03:04:38,000 GENERALLY USING BUOYANCY FORCE. 4081 03:04:38,000 --> 03:04:42,280 THE IDEA IF WE FUNCTIONALIZE 4082 03:04:42,280 --> 03:04:44,680 THOSE BEADS WITH CAPTURING 4083 03:04:44,680 --> 03:04:47,080 MATERIALS THAT CAN SPECIFICALLY 4084 03:04:47,080 --> 03:04:49,040 CAPTURE THE DNA THAT I WANT OR 4085 03:04:49,040 --> 03:04:51,920 THE TOTAL DNA THAT I WANT, SO IT 4086 03:04:51,920 --> 03:04:55,720 CAN CARRY THIS DNA, FLOAT TO THE 4087 03:04:55,720 --> 03:05:01,120 TOP, SO THEN WE CAN USE THAT 4088 03:05:01,120 --> 03:05:03,600 MATERIAL TO SEPARATE FIRSTLY 4089 03:05:03,600 --> 03:05:04,800 LYSIS THE STEW, AND THEN 4090 03:05:04,800 --> 03:05:07,640 SEPARATE THE DNA THAT WE WANT. 4091 03:05:07,640 --> 03:05:12,240 AND THEN WE APPLY THEM TO THE 4092 03:05:12,240 --> 03:05:16,400 LATERAL FLOW ASSAY, WHICH IS WE 4093 03:05:16,400 --> 03:05:18,600 SEE THE PREGNANCY TEST BASED 4094 03:05:18,600 --> 03:05:24,600 LIKE STRIPS, THAT WE CAN SAY IF 4095 03:05:24,600 --> 03:05:27,560 THIS EXISTS OR NOT, RIGHT? 4096 03:05:27,560 --> 03:05:31,280 HERE IS THE RESULT. 4097 03:05:31,280 --> 03:05:32,560 BY APPLYING THOSE BIO-SEPARATION 4098 03:05:32,560 --> 03:05:34,360 METHOD WE SEPARATE DNA FROM 4099 03:05:34,360 --> 03:05:38,280 STEW, AND THEN WE SPIKE IN THE 4100 03:05:38,280 --> 03:05:41,920 PATHOGENS INTO THE STEW, AND 4101 03:05:41,920 --> 03:05:42,680 THEN WITH DIFFERENT 4102 03:05:42,680 --> 03:05:44,680 CONCENTRATIONS FROM LOW TO HIGH, 4103 03:05:44,680 --> 03:05:46,640 I WANT TO SEE LIMIT OF DETECTION 4104 03:05:46,640 --> 03:05:52,480 OF THIS METHOD AND WE ALSO 4105 03:05:52,480 --> 03:05:54,360 QUANTIFYING INTENSITY OF THE 4106 03:05:54,360 --> 03:05:56,240 TEST DOTS HERE, LATER WE MADE 4107 03:05:56,240 --> 03:05:57,680 THE LINE. 4108 03:05:57,680 --> 03:06:01,800 SO THE TEST DOT HERE, WE HAVE 4109 03:06:01,800 --> 03:06:03,720 QUITE A LINEAR REGRESSION ON 4110 03:06:03,720 --> 03:06:04,560 THAT. 4111 03:06:04,560 --> 03:06:08,800 IF WE INCREASE, KEEP INCREASE 4112 03:06:08,800 --> 03:06:10,400 THE CONCENTRATION OF THE 4113 03:06:10,400 --> 03:06:15,760 PATHOGEN, YOU WILL BE LIKE FLAT. 4114 03:06:15,760 --> 03:06:19,880 SO LATER ON WE TRIED, ALSO TRIED 4115 03:06:19,880 --> 03:06:23,400 SENSITIVITY AND SELECTIVITY OF 4116 03:06:23,400 --> 03:06:24,280 OUR METHOD. 4117 03:06:24,280 --> 03:06:26,680 THE PICTURE ON THE RIGHT IS THE 4118 03:06:26,680 --> 03:06:27,680 TEST. 4119 03:06:27,680 --> 03:06:35,040 WE USED THE RPA AMPLIFICATION 4120 03:06:35,040 --> 03:06:39,960 METHOD TO AMPLIFY THE 4121 03:06:39,960 --> 03:06:42,200 CRYPTOSPOREDIAN, IF WE SPIKE 4122 03:06:42,200 --> 03:06:46,760 DIFFERENT OTHER BACTERIAS AND 4123 03:06:46,760 --> 03:06:50,320 PATHOGENS, PARASITES IN THERE, 4124 03:06:50,320 --> 03:06:53,320 GRDSM, WHATEVER, NO SIGNALS, 4125 03:06:53,320 --> 03:06:56,600 THAT'S THE SENSITIVITY AND 4126 03:06:56,600 --> 03:06:57,560 SELECTIVITY. 4127 03:06:57,560 --> 03:06:59,120 WE THINK, OKAY, TEST ONE STEW 4128 03:06:59,120 --> 03:07:02,400 SAMPLE, TELL US MORE 4129 03:07:02,400 --> 03:07:03,720 INFORMATION, NOT CHRIS CRYPTO, 4130 03:07:03,720 --> 03:07:07,560 BUT ALSO LIKE C. DIFF, E. COLI, 4131 03:07:07,560 --> 03:07:09,520 WHATEVER, THAT WE WANT, WE WANT 4132 03:07:09,520 --> 03:07:11,440 A MULTIPLEX. 4133 03:07:11,440 --> 03:07:18,600 SO THE PICTURE AT THE LEFT IS WE 4134 03:07:18,600 --> 03:07:22,000 ACTUALLY HAVE DIFFERENT MASTER 4135 03:07:22,000 --> 03:07:24,720 MIX OF THE RPA AMPLIFICATION, 4136 03:07:24,720 --> 03:07:27,680 AND THEN WE AMPLIFY TWO OR THREE 4137 03:07:27,680 --> 03:07:29,760 PATHOGENS AT A TIME. 4138 03:07:29,760 --> 03:07:35,240 SO WE CAN HAVE THREE MULTIPLEX 4139 03:07:35,240 --> 03:07:45,520 LIKEY MULTIPLEX 4140 03:07:46,280 --> 03:07:49,440 GIARDIA, THREE AT ONE TIME. 4141 03:07:49,440 --> 03:07:51,120 WE TRY DIFFERENT COMBINATIONS 4142 03:07:51,120 --> 03:07:54,920 AND SEE SELECTIVITY IS PRETTY 4143 03:07:54,920 --> 03:07:55,120 GOOD. 4144 03:07:55,120 --> 03:07:57,320 SO, LATER ON, WE ALSO HAVE 4145 03:07:57,320 --> 03:07:58,240 ANOTHER QUESTION. 4146 03:07:58,240 --> 03:08:02,120 SO, THE TEST STRIP LIKE THE 4147 03:08:02,120 --> 03:08:03,480 PREGNANCY TEST STRIP, BECAUSE OF 4148 03:08:03,480 --> 03:08:05,520 THE COVID MAYBE EVERYONE USING 4149 03:08:05,520 --> 03:08:08,920 THAT FOR COVID TEST, SO IT'S 4150 03:08:08,920 --> 03:08:09,720 PRETTY SHORT. 4151 03:08:09,720 --> 03:08:14,000 AND WE TRY TO HAVE LIKE TEN OR 4152 03:08:14,000 --> 03:08:16,560 FIVE PATHOGENS AT A TIME FOR ONE 4153 03:08:16,560 --> 03:08:18,880 STRIP BUT DOESN'T WORK. 4154 03:08:18,880 --> 03:08:21,200 SO WE WANT TO TRY LIKE AS MUCH 4155 03:08:21,200 --> 03:08:24,800 AS WE CAN, LIKE WE WANT TO GET 4156 03:08:24,800 --> 03:08:26,000 ONE SAMPLE AND GET AS MUCH 4157 03:08:26,000 --> 03:08:28,200 ANSWER AS WE CAN. 4158 03:08:28,200 --> 03:08:31,440 SO WE TRIED ANOTHER PLATFORM, 4159 03:08:31,440 --> 03:08:33,120 CALLED THIS PAPER ORIGAMI 4160 03:08:33,120 --> 03:08:33,640 MULTIPLEX SENSOR. 4161 03:08:33,640 --> 03:08:38,800 SO WHAT WE DO IS WE CHANGE THE 4162 03:08:38,800 --> 03:08:41,000 IDEA, THE LOGIC BEHIND IT. 4163 03:08:41,000 --> 03:08:45,360 LATERAL FLOW IN THE LATERAL, AND 4164 03:08:45,360 --> 03:08:47,000 WHAT IF WE USE MICRO FLUID NOT 4165 03:08:47,000 --> 03:08:50,520 ONLY ON LATERAL BUT VERTICAL, 4166 03:08:50,520 --> 03:08:50,960 OKAY? 4167 03:08:50,960 --> 03:08:54,120 WE HAVE PRINT THOSE PATTERNS ON 4168 03:08:54,120 --> 03:08:56,720 A CHROMATOGRAPHY PAPER, AND THEN 4169 03:08:56,720 --> 03:08:59,680 WHAT YOU SEE HERE IN THE BLACK 4170 03:08:59,680 --> 03:09:03,320 COLOR ARE WAX, SO WAX ARE 4171 03:09:03,320 --> 03:09:06,040 HYDROPHOBIC, SO IF YOU HAVE A 4172 03:09:06,040 --> 03:09:08,240 LIQUID SAMPLE APPLIED TO THE 4173 03:09:08,240 --> 03:09:11,800 PAPER, STUCK TO THE PAPER, AND 4174 03:09:11,800 --> 03:09:13,840 TRAVELING FROM LAYER 1 TO 2 TO 3 4175 03:09:13,840 --> 03:09:16,120 TO 4. 4176 03:09:16,120 --> 03:09:19,240 AND ALL THE WAY DOWN, IT TRAVELS 4177 03:09:19,240 --> 03:09:21,400 LIKE NOT VERTICALLY BUT -- NOT 4178 03:09:21,400 --> 03:09:24,560 LATERALLY BUT VERTICALLY, RIGHT? 4179 03:09:24,560 --> 03:09:28,400 SO, WE VIEWED THIS KIND OF 4180 03:09:28,400 --> 03:09:29,160 VERTICAL ORIGAMI SENSOR BECAUSE 4181 03:09:29,160 --> 03:09:31,680 AT THE VERY BEGINNING WE PRINT 4182 03:09:31,680 --> 03:09:34,960 AS A PAPER, THEY WOULD FOLD IT 4183 03:09:34,960 --> 03:09:39,640 TO A SENSOR. 4184 03:09:39,640 --> 03:09:42,160 SO, THEN WE APPLIED THE SAMPLE 4185 03:09:42,160 --> 03:09:43,480 AT THE VERY FIRST LAYER, AND 4186 03:09:43,480 --> 03:09:45,000 WE'LL GO DOWN, DOWN, DOWN, DOWN, 4187 03:09:45,000 --> 03:09:48,440 ALL THE WAY TO THE BOTTOM, AND 4188 03:09:48,440 --> 03:09:54,880 IF WE HAVE THE C. DIFF APPEAR IT 4189 03:09:54,880 --> 03:09:56,080 WILL TURN COLOR, FLUORESCENT 4190 03:09:56,080 --> 03:09:59,240 GREEN, IF THERE'S NO, IT'S AN 4191 03:09:59,240 --> 03:10:02,000 ACTIVE CONTROL, BROWNISH. 4192 03:10:02,000 --> 03:10:02,560 OKAY? 4193 03:10:02,560 --> 03:10:06,040 SO, BY DOING THAT, WE CAN DO 4194 03:10:06,040 --> 03:10:07,240 EIGHT PATHOGENS AT A TIME 4195 03:10:07,240 --> 03:10:09,240 BECAUSE LOOK AT THE PATTERN IN 4196 03:10:09,240 --> 03:10:14,040 THE MIDDLE, SO EACH ARM IS ONE 4197 03:10:14,040 --> 03:10:14,480 PATHOGEN. 4198 03:10:14,480 --> 03:10:16,320 SO ONE SAMPLE INJECT IN THE 4199 03:10:16,320 --> 03:10:17,920 MIDDLE, WILL TRAVEL DOWN AND 4200 03:10:17,920 --> 03:10:20,080 SPREAD AND DOWN TO THE BOTTOM, 4201 03:10:20,080 --> 03:10:24,960 AND WE CAN GET EIGHT ANSWERS AT 4202 03:10:24,960 --> 03:10:26,520 A TIME. 4203 03:10:26,520 --> 03:10:27,600 ACCURACY AND SENSITIVITY OVER 4204 03:10:27,600 --> 03:10:28,680 98% FOR NOW. 4205 03:10:28,680 --> 03:10:31,880 WE'RE GOING TO IMPROVE THAT. 4206 03:10:31,880 --> 03:10:40,960 SO RIGHT NOW WE TRIED C. DIFF, 4207 03:10:40,960 --> 03:10:44,240 SHIGELLA, ETEC, EPEC, ROTAVIRUS, 4208 03:10:44,240 --> 03:10:54,480 NOROVIRUS, TYPE 1 AND 2, 4209 03:10:54,480 --> 03:10:56,600 CRYPTOSPORIDUM, GIARDIA. 4210 03:10:56,600 --> 03:11:02,800 ALL OF THOSE LIKE THIS IS -- WE 4211 03:11:02,800 --> 03:11:03,800 RECENTLY GOT THE PATENT ISSUED, 4212 03:11:03,800 --> 03:11:08,280 GLOBAL PATENT FOR THIS PAPER 4213 03:11:08,280 --> 03:11:10,160 ORIGAMI AND MICRO SPHERE TWO 4214 03:11:10,160 --> 03:11:11,040 YEARS AGO. 4215 03:11:11,040 --> 03:11:15,280 AND I AM THE PRIMARY MENTOR FOR 4216 03:11:15,280 --> 03:11:18,160 BOTH OF THE PAT -- PRIMARY 4217 03:11:18,160 --> 03:11:19,440 INVENTOR FOR BOTH OF THE 4218 03:11:19,440 --> 03:11:22,080 PATENTS, AND THAT IS MY 4219 03:11:22,080 --> 03:11:23,240 RESEARCH. 4220 03:11:23,240 --> 03:11:24,600 [APPLAUSE] 4221 03:11:24,600 --> 03:11:27,040 4222 03:11:27,040 --> 03:11:29,440 >> VERY CLEVER IDEAS. 4223 03:11:29,440 --> 03:11:31,200 I LOVE THEM. 4224 03:11:31,200 --> 03:11:33,280 FOURTH PANEL, LAST SLIDE, 4225 03:11:33,280 --> 03:11:34,800 INTERMEDIATE POSITIONS. 4226 03:11:34,800 --> 03:11:35,240 WHAT ARE THOSE FOR? 4227 03:11:35,240 --> 03:11:36,760 >> THAT'S A GOOD QUESTION, FOR 4228 03:11:36,760 --> 03:11:39,360 THE TWO DOTS, RIGHT? 4229 03:11:39,360 --> 03:11:43,360 SO, THE VERY FIRST IDEA FOR THAT 4230 03:11:43,360 --> 03:11:47,840 WAS TO INCUBATE, YOU KNOW, LIKE 4231 03:11:47,840 --> 03:11:49,800 EITHER FOR RPA OR LAMP 4232 03:11:49,800 --> 03:11:53,520 APPLICATION REQUIRES A HEATING. 4233 03:11:53,520 --> 03:11:57,880 EITHER TO 37 DEGREES C OR OVER 4234 03:11:57,880 --> 03:11:59,440 40 DEGREES C. 4235 03:11:59,440 --> 03:12:02,960 SO WE HAVE LIKE THICKER, YOU SEE 4236 03:12:02,960 --> 03:12:05,760 A THICKER DESIGN RATHER THAN ALL 4237 03:12:05,760 --> 03:12:08,200 THE OTHER LAYERS BECAUSE IT ACTS 4238 03:12:08,200 --> 03:12:10,960 AS THE RESERVOIR TO KEEP SOME OF 4239 03:12:10,960 --> 03:12:13,000 THE SAMPLES OVER THERE, AND WE 4240 03:12:13,000 --> 03:12:14,560 HEAT UP. 4241 03:12:14,560 --> 03:12:17,520 WE USE -- ACTUALLY AT THE VERY 4242 03:12:17,520 --> 03:12:19,160 BEGINNING WE TRIED THE FOOT 4243 03:12:19,160 --> 03:12:20,880 WARMER YOU PUT IN YOUR SHOES IN 4244 03:12:20,880 --> 03:12:23,960 THE WINTER TO KEEP YOU WARM. 4245 03:12:23,960 --> 03:12:28,000 AND PUT THAT LAYER UNDERNEATH 4246 03:12:28,000 --> 03:12:28,200 THAT. 4247 03:12:28,200 --> 03:12:29,760 WHEN YOU SQUEEZE IT, IT WOULD 4248 03:12:29,760 --> 03:12:33,480 GENERATE HEAT AND YOU REMOVE IT, 4249 03:12:33,480 --> 03:12:36,640 DRAIN DOWN SO SHOWS COLOR AND IT 4250 03:12:36,640 --> 03:12:38,320 REMAINS LOW COST AND ALSO CAN 4251 03:12:38,320 --> 03:12:40,120 HEAT IT UP. 4252 03:12:40,120 --> 03:12:42,560 LATER ON BECAUSE OF TECHNOLOGY 4253 03:12:42,560 --> 03:12:47,160 GOT MUCH BETTER, WE CAN DO A 4254 03:12:47,160 --> 03:12:48,440 ROOM TEMPERATURE APPLICATION, NO 4255 03:12:48,440 --> 03:12:52,720 NEED FOR HEATING AT THIS MOMENT. 4256 03:12:52,720 --> 03:12:55,440 BUT FOR ESPECIALLY LAMP, THERE'S 4257 03:12:55,440 --> 03:12:58,400 A ROOM TEMPERATURE LAMP, 4258 03:12:58,400 --> 03:12:59,640 REALTIME ROOM TEMPERATURE LAMP, 4259 03:12:59,640 --> 03:13:05,080 SO WE CAN APPLY THAT AND WE HAVE 4260 03:13:05,080 --> 03:13:07,280 FURTHER RESEARCH DOING OTHER 4261 03:13:07,280 --> 03:13:10,680 AMPLIFICATIONS THAT IS UNDER THE 4262 03:13:10,680 --> 03:13:14,720 PATENT REVIEW SO I CANNOT REVIEW 4263 03:13:14,720 --> 03:13:15,160 HERE. 4264 03:13:15,160 --> 03:13:18,200 SO, YEAH, THE TECHNOLOGY IS 4265 03:13:18,200 --> 03:13:18,800 MOVING FORWARD. 4266 03:13:18,800 --> 03:13:19,120 >> AGREED. 4267 03:13:19,120 --> 03:13:20,640 THIS IS REALLY COOL. 4268 03:13:20,640 --> 03:13:24,240 CURIOUS HOW TEMPERATURE STABLE 4269 03:13:24,240 --> 03:13:26,640 ARE THESE, LIKE LOW-RESOURCE 4270 03:13:26,640 --> 03:13:27,440 SETTINGS WITHOUT REFRIGERATION, 4271 03:13:27,440 --> 03:13:28,480 TEMPERATURE UP TO HOW LONG? 4272 03:13:28,480 --> 03:13:31,040 YOU SAID THIS WAS WAX? 4273 03:13:31,040 --> 03:13:32,080 CURIOUS ABOUT THAT. 4274 03:13:32,080 --> 03:13:38,360 >> SO THE WAX -- SO, WHEN WE 4275 03:13:38,360 --> 03:13:39,600 MADE THIS, WE ONLY PRINT WAX ON 4276 03:13:39,600 --> 03:13:41,640 ONE SIDE OF THE PAPER AND THEN 4277 03:13:41,640 --> 03:13:46,520 WE PUT IN THE OVEN AND MAKE THE 4278 03:13:46,520 --> 03:13:49,880 WAX WET AND IT WILL SINK THROUGH 4279 03:13:49,880 --> 03:13:53,160 THE PAPER, SO GENERATE ACTUALLY 4280 03:13:53,160 --> 03:13:56,000 THE REAL HYDROPHOBIC WALL IN THE 4281 03:13:56,000 --> 03:13:56,320 PAPER. 4282 03:13:56,320 --> 03:13:58,840 AND THEN THAT WILL -- THAT IS 4283 03:13:58,840 --> 03:14:03,520 OVER 100 DEGREES C, SO I DON'T 4284 03:14:03,520 --> 03:14:05,280 THINK ANY LIKE ENVIRONMENTAL 4285 03:14:05,280 --> 03:14:07,920 TEMPERATURE WILL HEAT IT UP 4286 03:14:07,920 --> 03:14:10,520 AGAIN TO DAMAGE THIS CHANNEL. 4287 03:14:10,520 --> 03:14:15,000 BUT INCLUDING THE HEATING THAT I 4288 03:14:15,000 --> 03:14:16,440 JUST MENTIONED, EITHER 37 4289 03:14:16,440 --> 03:14:20,080 DEGREES C OR OVER 45, SO IT 4290 03:14:20,080 --> 03:14:22,040 WON'T DAMAGE THE PATTERN THAT WE 4291 03:14:22,040 --> 03:14:25,400 DEVELOPED. 4292 03:14:25,400 --> 03:14:30,120 AND ALSO THAT'S A GOOD QUESTION 4293 03:14:30,120 --> 03:14:31,120 TOO, YOU SEE THIS TRANSPARENT 4294 03:14:31,120 --> 03:14:32,320 THING OVER HERE? 4295 03:14:32,320 --> 03:14:38,000 IT'S THE MEMBRANE THAT WE TRY TO 4296 03:14:38,000 --> 03:14:41,200 SEAL EVERYTHING INTO A SEALED 4297 03:14:41,200 --> 03:14:43,280 DEVICE THAT LIKE ONCE YOU INJECT 4298 03:14:43,280 --> 03:14:45,680 THERE'S ANOTHER SEAL AT THE TOP, 4299 03:14:45,680 --> 03:14:50,040 SO THE USER WON'T TOUCH THE STEW 4300 03:14:50,040 --> 03:14:52,320 BECAUSE, YOU KNOW, THEY APPLY 4301 03:14:52,320 --> 03:15:01,840 THE STEW SAMPLES, TO KEEP IT 4302 03:15:01,840 --> 03:15:02,120 SANITIZED. 4303 03:15:02,120 --> 03:15:05,520 >> SO, I SEE THAT LIKE THE 4304 03:15:05,520 --> 03:15:08,840 NEGATIVE CONTROL C. DIFF SHOW 4305 03:15:08,840 --> 03:15:10,240 GREEN FLUORESCENCE, WHAT ABOUT 4306 03:15:10,240 --> 03:15:14,520 OTHER SPECIES, DO THEY SHOW 4307 03:15:14,520 --> 03:15:15,600 DIFFERENT COLOR FLUORESCENCE ON 4308 03:15:15,600 --> 03:15:16,480 DIFFERENT PATTERN? 4309 03:15:16,480 --> 03:15:18,880 HOW CAN YOU TELL? 4310 03:15:18,880 --> 03:15:20,640 >> YEAH, FOR THIS, YES, THAT'S 4311 03:15:20,640 --> 03:15:21,640 A GREAT QUESTION. 4312 03:15:21,640 --> 03:15:25,240 SO AT THE VERY BOTTOM YOU SEE 4313 03:15:25,240 --> 03:15:27,760 THE -- YEAH, YOU SEE THE PICTURE 4314 03:15:27,760 --> 03:15:28,440 RIGHT HERE, RIGHT? 4315 03:15:28,440 --> 03:15:31,840 SO, ONE OF THE DOTS, ONE OF THE 4316 03:15:31,840 --> 03:15:34,680 TWO DOTS IS THE C. DIFF. 4317 03:15:34,680 --> 03:15:38,120 IT WILL BE THIS ONE OUTSIDE, 4318 03:15:38,120 --> 03:15:40,600 OUTSIDE IS CONTROL, INSIDE IS A 4319 03:15:40,600 --> 03:15:40,920 TEST. 4320 03:15:40,920 --> 03:15:43,880 WE WILL LABEL WHICH ARM IS WHICH 4321 03:15:43,880 --> 03:15:45,840 AT THE VERY BOTTOM. 4322 03:15:45,840 --> 03:15:48,360 SO THEY ESSENTIALLY HAVE THE 4323 03:15:48,360 --> 03:15:50,200 SAME SIGNAL BUT BECAUSE WE LABEL 4324 03:15:50,200 --> 03:15:54,920 IT, YOU WILL KNOW WHICH ARM 4325 03:15:54,920 --> 03:15:55,400 REPRESENTS WHAT. 4326 03:15:55,400 --> 03:15:55,800 >> THANK YOU. 4327 03:15:55,800 --> 03:15:59,160 >> YES. 4328 03:15:59,160 --> 03:16:01,720 >> THANK YOU SO MUCH. 4329 03:16:01,720 --> 03:16:08,800 >> ALL RIGHT, THANK YOU. 4330 03:16:08,800 --> 03:16:09,080 [APPLAUSE] 4331 03:16:09,080 --> 03:16:11,720 >> NEXT SPEAKER IS DR. ETHAN 4332 03:16:11,720 --> 03:16:15,400 COHEN FROM OFFICE OF SCIENCE AND 4333 03:16:15,400 --> 03:16:18,080 ENGINEERING LABORATORY AT FDA. 4334 03:16:18,080 --> 03:16:23,880 4335 03:16:23,880 --> 03:16:28,320 >> I'D LIKE TO THANK THE 4336 03:16:28,320 --> 03:16:29,520 ORGANIZERS FOR INVITING ME. 4337 03:16:29,520 --> 03:16:35,600 I'D LIKE TO TALK TODAY ABOUT -- 4338 03:16:35,600 --> 03:16:41,600 COBALT CAUSES RAPID CHANGE IN 4339 03:16:41,600 --> 03:16:43,600 RAPID STRUCTURE, OF THE RABBIT 4340 03:16:43,600 --> 03:16:43,960 RETINA. 4341 03:16:43,960 --> 03:16:48,840 I WORK AT FDA SO WE HAVE THIS 4342 03:16:48,840 --> 03:16:49,160 DISCLAIMER. 4343 03:16:49,160 --> 03:16:50,040 GO PAST THAT. 4344 03:16:50,040 --> 03:16:52,800 SO I WORK AT MEDICAL DEVICE 4345 03:16:52,800 --> 03:16:55,280 COMPANY, MEDICAL DEVICE 4346 03:16:55,280 --> 03:16:57,360 GOVERNMENT AGENCY. 4347 03:16:57,360 --> 03:17:00,760 AND WE'RE ALWAYS DEALING WITH 4348 03:17:00,760 --> 03:17:03,520 TOXICOLOGY ISSUES, TOXICOLOGY OF 4349 03:17:03,520 --> 03:17:05,040 DRUGS, TOXICOLOGY OF DEVICES, 4350 03:17:05,040 --> 03:17:07,680 SOME DEVICES WORK WELL, SOME 4351 03:17:07,680 --> 03:17:08,840 DEVICES FAIL. 4352 03:17:08,840 --> 03:17:10,840 AS DAN POINTED OUT, WE NEED TO 4353 03:17:10,840 --> 03:17:14,680 DEVELOP A SERIES OF REGULATORY 4354 03:17:14,680 --> 03:17:17,800 SCIENCE TOOLS TO HELP INDUSTRY 4355 03:17:17,800 --> 03:17:20,920 BRAIN DEVICES THROUGH THEY CAN 4356 03:17:20,920 --> 03:17:23,560 DEMONSTRATE ARE SAFE, REDUCES 4357 03:17:23,560 --> 03:17:25,720 USE OF ANIMALS TO SOME DEGREE. 4358 03:17:25,720 --> 03:17:29,240 THREE, THAT GIVES SOME IDEA OF 4359 03:17:29,240 --> 03:17:31,200 HOW THESE SUBSTANCES ARE 4360 03:17:31,200 --> 03:17:34,400 AFFECTING LIVING TISSUE IN REAL 4361 03:17:34,400 --> 03:17:35,280 TIME. 4362 03:17:35,280 --> 03:17:38,200 SO, IN THE CASE OF -- I'M GOING 4363 03:17:38,200 --> 03:17:40,960 TO TALK ABOUT THE CASE OF 4364 03:17:40,960 --> 03:17:44,040 COBALT, AND THERE WERE ABOUT A 4365 03:17:44,040 --> 03:17:49,080 MILLION COBALT CHROMIUM HIP 4366 03:17:49,080 --> 03:17:50,280 IMPLANTS PRODUCED, AND A 4367 03:17:50,280 --> 03:17:52,600 SUBSTANTIAL NUMBER OF THESE 4368 03:17:52,600 --> 03:17:54,760 IMPLANTS PRODUCE HIGH LEVELS OF 4369 03:17:54,760 --> 03:17:56,080 COBALT IN THE PATIENT'S BLOOD, 4370 03:17:56,080 --> 03:18:01,760 AND IN A VERY LIMITED SUBSET OF 4371 03:18:01,760 --> 03:18:04,480 PEOPLE THIS LEVEL OF COBALT WAS 4372 03:18:04,480 --> 03:18:13,240 ENOUGH TO IMPAIR VISION OR CAUSE 4373 03:18:13,240 --> 03:18:16,520 TINNITUS OR BALANCE ISSUES. 4374 03:18:16,520 --> 03:18:18,480 WHAT ARE THE MECHANISMS OF 4375 03:18:18,480 --> 03:18:21,800 COBALT TOXICITY AND I'M USING 4376 03:18:21,800 --> 03:18:24,080 THE LIVING RETINA AS THE MODEL 4377 03:18:24,080 --> 03:18:26,920 SYSTEM TO EXAMINE THESE EFFECTS 4378 03:18:26,920 --> 03:18:28,440 IN REAL TIME. 4379 03:18:28,440 --> 03:18:38,320 AND I USE A THING CALLED OP -- 4380 03:18:38,320 --> 03:18:39,320 OPTICAL TOMOGRAPHY. 4381 03:18:39,320 --> 03:18:41,840 COBALT HAS MANY EFFECTS ON 4382 03:18:41,840 --> 03:18:49,840 TISSUES, ONE WE DON'T UNDERSTAND 4383 03:18:49,840 --> 03:18:51,040 WELL ETHNICITY ALLOTHIONINE 4384 03:18:51,040 --> 03:18:53,480 RECEPTOR ACTIVATION, OF INTEREST 4385 03:18:53,480 --> 03:18:57,280 TO MITOCHONDRIAL PEOPLE BY 4386 03:18:57,280 --> 03:18:58,040 INTERFERING WITH REDOX SYSTEMS 4387 03:18:58,040 --> 03:19:03,520 AND WE KNOW IT CAN MIMIC 4388 03:19:03,520 --> 03:19:05,640 BACTERIAL CELL WALL ACTIVATING 4389 03:19:05,640 --> 03:19:07,680 TOLL 4 RECEPTORS, COBALT AND 4390 03:19:07,680 --> 03:19:09,040 NICKEL BOTH. 4391 03:19:09,040 --> 03:19:10,880 AND IN ADDITION, WHAT YOU MAY 4392 03:19:10,880 --> 03:19:20,640 NOT KNOW IS COBALT ACTIVATES 4393 03:19:20,640 --> 03:19:21,400 PHOSPHO-DIESTERASE. 4394 03:19:21,400 --> 03:19:23,920 IT BINDS TO PROTEIN ON THE 4395 03:19:23,920 --> 03:19:26,400 HYPOXIA INDUCED FACTOR THAT 4396 03:19:26,400 --> 03:19:28,760 MIMICS HYPOXIA, AND THIS CAUSES 4397 03:19:28,760 --> 03:19:30,480 CHANGES IN GENE TRANSCRIPTION. 4398 03:19:30,480 --> 03:19:32,120 AND FINALLY BEING A 4399 03:19:32,120 --> 03:19:35,240 PHYSIOLOGIST, YOU KNOW THE 4400 03:19:35,240 --> 03:19:37,040 COBALT INHIBITS CALCIUM 4401 03:19:37,040 --> 03:19:37,720 CHANNELS. 4402 03:19:37,720 --> 03:19:39,120 SO, WHAT HAPPENS TO A PATIENT 4403 03:19:39,120 --> 03:19:44,480 WHO HAS A VERY SEVERE CASE OF 4404 03:19:44,480 --> 03:19:44,880 COBALTERIA? 4405 03:19:44,880 --> 03:19:49,880 THEY COULD HAVE A LEVEL OF 900 4406 03:19:49,880 --> 03:19:52,720 MICROGRAMS PER ML, AND WHAT YOU 4407 03:19:52,720 --> 03:19:54,680 CAN SEE HERE OCT IMAGE WHICH DAN 4408 03:19:54,680 --> 03:19:57,800 HAS DISCUSSED, THIS IS WHAT A 4409 03:19:57,800 --> 03:19:59,160 MORE NORMAL CLINICAL OCT IMAGES 4410 03:19:59,160 --> 03:20:01,560 LOOKS LIKE OF A RETINA THAN 4411 03:20:01,560 --> 03:20:02,800 EARLIER GENERATION, AND LET'S 4412 03:20:02,800 --> 03:20:05,200 SEE, DOES THIS HAVE A LASER 4413 03:20:05,200 --> 03:20:12,720 POINTER? 4414 03:20:12,720 --> 03:20:17,440 4415 03:20:17,440 --> 03:20:18,200 OVER HERE? 4416 03:20:18,200 --> 03:20:26,320 HERE, I CAN USE THIS I GUESS. 4417 03:20:26,320 --> 03:20:34,840 WHAT YOU CAN SEE, EXCUSE ME, 4418 03:20:34,840 --> 03:20:42,200 WHAT YOU CAN SEE HERE, THE UPPER 4419 03:20:42,200 --> 03:20:43,480 LEFT-HAND PANEL A 4420 03:20:43,480 --> 03:20:45,600 CROSS-SECTIONAL IMAGE USING A 4421 03:20:45,600 --> 03:20:52,440 NORMAL CLINICAL OCT, FOVEA, 4422 03:20:52,440 --> 03:20:53,760 THERE'S A LOT OF GANGLION CELLS 4423 03:20:53,760 --> 03:20:55,320 BUT YOU CAN'T SEE A LOT WHAT'S 4424 03:20:55,320 --> 03:20:58,200 GOING ON IN THE PHOTORECEPTORS. 4425 03:20:58,200 --> 03:21:00,920 THESE PATIENTS WHEN THEY COME 4426 03:21:00,920 --> 03:21:03,200 INTO THE CLINIC IF YOU LOOK AT 4427 03:21:03,200 --> 03:21:04,720 VISUAL FIELDS, WHICH YOU CAN SEE 4428 03:21:04,720 --> 03:21:08,200 ON THE FAR RIGHT, YOU CAN SEE 4429 03:21:08,200 --> 03:21:13,920 VERY LARGE REGION, WHERE THEY 4430 03:21:13,920 --> 03:21:22,000 LOST CENTRAL VISION. 4431 03:21:22,000 --> 03:21:32,680 AND ALSO WHEN YOU PROBE THE 4432 03:21:32,680 --> 03:21:33,920 RETINA USING MULTI-FOCAL 4433 03:21:33,920 --> 03:21:35,160 ELECTRORETINOGRAMS REFLECTING 4434 03:21:35,160 --> 03:21:36,000 LARGELY OUTER RETINAL FUNCTION, 4435 03:21:36,000 --> 03:21:37,560 THIS IS WHAT A NORMAL RETINA 4436 03:21:37,560 --> 03:21:40,080 LOOKS LIKE WHEN YOU PROBE THE 4437 03:21:40,080 --> 03:21:42,320 SURFACE OF THE FUNDUS, SENSITIVE 4438 03:21:42,320 --> 03:21:45,400 IN OUR FOVEA, YOU SEE A PEAK OF 4439 03:21:45,400 --> 03:21:47,080 SENSITIVITY IN PATIENTS, THIS 4440 03:21:47,080 --> 03:21:48,480 PEAK IS COMPLETELY LOST. 4441 03:21:48,480 --> 03:21:52,560 AND THIS SUGGESTS THAT THERE'S A 4442 03:21:52,560 --> 03:21:58,480 SIGNIFICANT DEFICIT TO THE 4443 03:21:58,480 --> 03:21:59,080 PHOTORECEPTORS. 4444 03:21:59,080 --> 03:22:01,840 SO, LET'S SEE. 4445 03:22:01,840 --> 03:22:02,040 OKAY. 4446 03:22:02,040 --> 03:22:05,440 SO, WHAT WE'VE -- I'VE USED HERE 4447 03:22:05,440 --> 03:22:09,080 IS A VERY SPECIAL PREPARATION 4448 03:22:09,080 --> 03:22:11,160 WHERE WE CAN KEEP THE RETINA 4449 03:22:11,160 --> 03:22:13,880 ALIVE OUTSIDE THE BODY ATTACHED 4450 03:22:13,880 --> 03:22:18,920 TO PIGMENT EPITHELIUM, THIS IS 4451 03:22:18,920 --> 03:22:21,080 CALLED A RABBIT EYE CUP 4452 03:22:21,080 --> 03:22:21,640 PREPARATION HERE. 4453 03:22:21,640 --> 03:22:24,960 BASICALLY THE BACK HALF OF THE 4454 03:22:24,960 --> 03:22:26,880 RABBIT EYE IS EVERTED, LIKE A 4455 03:22:26,880 --> 03:22:28,760 BALL CUT IN HALF, EVERTED AND 4456 03:22:28,760 --> 03:22:30,960 MOUNTED ON THE SORT OF DOME 4457 03:22:30,960 --> 03:22:33,480 CHAMBER, AND THEN USING OPTICAL 4458 03:22:33,480 --> 03:22:35,280 CURRENT STEMMOGRAPHY CAN SEE ALL 4459 03:22:35,280 --> 03:22:45,800 THE LAYERS OF THE LIVING RETINA 4460 03:22:48,760 --> 03:22:51,440 IN REAL TIME, HERE IS THE 4461 03:22:51,440 --> 03:22:57,800 GANGLION CELL LAYER, AND HERE, 4462 03:22:57,800 --> 03:22:59,040 THESE LAYERS HERE ARE 4463 03:22:59,040 --> 03:23:02,400 PHOTORECEPTOR ASSOCIATED LAYERS. 4464 03:23:02,400 --> 03:23:04,360 REMEMBER I SAID THE MULTI-FOCAL 4465 03:23:04,360 --> 03:23:05,240 ELECTRORETINOGRAM IS SEVERELY 4466 03:23:05,240 --> 03:23:08,280 LOST IN THESE PATIENTS THAT HAVE 4467 03:23:08,280 --> 03:23:09,520 HIGH COBALTERUA. 4468 03:23:09,520 --> 03:23:16,600 NOW WE LOOK AT OCT IMAGES OF 4469 03:23:16,600 --> 03:23:18,280 RABBIT RETINA CAN PERFUSE IN 4470 03:23:18,280 --> 03:23:20,160 REAL TIME AND SEE EFFECT ON THE 4471 03:23:20,160 --> 03:23:21,800 RETINAL STRUCTURE. 4472 03:23:21,800 --> 03:23:23,560 THAT'S REALLY THE ADVANTAGE HERE 4473 03:23:23,560 --> 03:23:25,720 OF THIS RETINAL PREPARATION, WE 4474 03:23:25,720 --> 03:23:26,960 DON'T HAVE ADAPTIVE OPTICS, SO 4475 03:23:26,960 --> 03:23:29,320 WE DON'T HAVE TO DEAL WITH THE 4476 03:23:29,320 --> 03:23:30,240 DISTORTIONS OF THE EYE, AND WE 4477 03:23:30,240 --> 03:23:34,360 DON'T HAVE TO DEAL WITH THE 4478 03:23:34,360 --> 03:23:38,960 PHARMACOKINETICS AND PARTITION 4479 03:23:38,960 --> 03:23:49,440 COEFFICIENTS OF DRUGS, LIVER 4480 03:23:51,960 --> 03:23:53,760 PATHWAYS, KIDNEY ALLUTIAN. 4481 03:23:53,760 --> 03:23:56,600 HERE WE SEE TEN MINUTES AFTER, 4482 03:23:56,600 --> 03:24:03,600 YOU CAN SEE CLEARLY HERE THAT 4483 03:24:03,600 --> 03:24:06,400 THERE'S A LOSS OF OF THE FINE 4484 03:24:06,400 --> 03:24:14,200 LINE NEXT TO THE PIGMENT 4485 03:24:14,200 --> 03:24:16,080 EPITHELIUM AND INNER 4486 03:24:16,080 --> 03:24:18,520 SEGMENT/OUTER SEGMENT LINE. 4487 03:24:18,520 --> 03:24:23,400 IN ADDITION YOU CAN SEE THERE'S 4488 03:24:23,400 --> 03:24:25,360 CHANGES ALSO THE MEMBRANE WHERE 4489 03:24:25,360 --> 03:24:28,200 THE GANGLION CELLS ARE. 4490 03:24:28,200 --> 03:24:30,200 SO WHAT THESE TWO IMAGES, THEY 4491 03:24:30,200 --> 03:24:31,200 ARE FLIPPED OF THE SAME RETINA 4492 03:24:31,200 --> 03:24:33,960 BEFORE AND AFTER THE DRUG SO CAN 4493 03:24:33,960 --> 03:24:35,000 YOU DIRECTLY COMPARE THE 4494 03:24:35,000 --> 03:24:35,440 STRUCTURE. 4495 03:24:35,440 --> 03:24:38,280 SO YOU CAN SEE THERE'S A 4496 03:24:38,280 --> 03:24:40,840 SIGNIFICANT CHANGE IN THE OUTER 4497 03:24:40,840 --> 03:24:44,960 RETINA, IT'S VERY RAPID, WHICH 4498 03:24:44,960 --> 03:24:45,720 IS SOMEWHAT UNEXPECTED, AND WHAT 4499 03:24:45,720 --> 03:24:50,240 WE'RE TRYING TO FIGURE OUT IS 4500 03:24:50,240 --> 03:24:51,960 WHAT IS CAUSING THIS 4501 03:24:51,960 --> 03:24:52,440 DEGENERATION. 4502 03:24:52,440 --> 03:24:54,360 SO, WHAT WE CAN DO NOW IS WE CAN 4503 03:24:54,360 --> 03:24:57,760 LOOK AT HOW THE CHANGES IN THAT 4504 03:24:57,760 --> 03:24:58,880 RETINAL STRUCTURE CHANGE WITH 4505 03:24:58,880 --> 03:25:06,000 TIME, AND WE DO THAT BY 4506 03:25:06,000 --> 03:25:09,040 TAKING -- LET'S SEE HERE. 4507 03:25:09,040 --> 03:25:10,600 IT'S NOT GOING BACKWARDS. 4508 03:25:10,600 --> 03:25:13,440 HOLD ON FOR A SECOND. 4509 03:25:13,440 --> 03:25:18,040 4510 03:25:18,040 --> 03:25:20,560 WE CAN TAKE A SQUARE REGION HERE 4511 03:25:20,560 --> 03:25:23,840 OF THE RETINA, AVERAGE IT 4512 03:25:23,840 --> 03:25:25,360 LATERALLY, AND THEN TAKE 4513 03:25:25,360 --> 03:25:26,520 DIFFERENT TIME POINTS EVERY 15 4514 03:25:26,520 --> 03:25:29,520 SECONDS OF THE CHANGES IN THE 4515 03:25:29,520 --> 03:25:31,480 RETINAL STRUCTURE WITH THE 4516 03:25:31,480 --> 03:25:32,040 PRESENCE OF COBALT. 4517 03:25:32,040 --> 03:25:36,320 AND SO HERE YOU SEE A TEST WHERE 4518 03:25:36,320 --> 03:25:40,000 WE USED NO COBALT AND YOU CAN 4519 03:25:40,000 --> 03:25:42,240 SEE HERE WHEN WE PUT THE RINGER 4520 03:25:42,240 --> 03:25:44,360 WHICH HAS NOTHING IN IT, 4521 03:25:44,360 --> 03:25:48,240 BASICALLY THERE'S VERY ALSO 4522 03:25:48,240 --> 03:25:49,600 EFFECT -- VERY LITTLE EFFECT ON 4523 03:25:49,600 --> 03:25:50,960 THE RETINAL STRUCTURE, THIS IS A 4524 03:25:50,960 --> 03:25:54,080 CROSS-SECTION AS A LINE OF THE 4525 03:25:54,080 --> 03:25:56,000 RETINAL STRUCTURE, BEFORE AND 4526 03:25:56,000 --> 03:25:56,960 AFTER THE COBALT RIGHT HERE AND 4527 03:25:56,960 --> 03:25:58,080 RIGHT THERE. 4528 03:25:58,080 --> 03:26:01,480 AND YOU CAN SEE BASICALLY 4529 03:26:01,480 --> 03:26:03,120 THERE'S VERY LITTLE EFFECT. 4530 03:26:03,120 --> 03:26:08,400 NOW, IF WE DO THE SAME THING, 4531 03:26:08,400 --> 03:26:12,000 EXCUSE ME, IN THE PRESENCE OF 4532 03:26:12,000 --> 03:26:15,000 COBALT YOU CAN SEE HERE THAT 4533 03:26:15,000 --> 03:26:17,880 THERE'S A DROP IN THE INNER 4534 03:26:17,880 --> 03:26:19,320 SEGMENT, OUTER SEGMENT LINE AND 4535 03:26:19,320 --> 03:26:29,840 A DROP IN WHAT WE KNOW ARE THE 4536 03:26:30,440 --> 03:26:30,960 INTERDIGITATION ZONE, ON THE 4537 03:26:30,960 --> 03:26:32,120 ORDER OF TEN MINUTES. 4538 03:26:32,120 --> 03:26:35,000 IF THIS WAS ACTING THROUGH SOME 4539 03:26:35,000 --> 03:26:36,640 SORT OF HYPOXIA-INDUCED FACTOR 4540 03:26:36,640 --> 03:26:38,360 OR GENE TRANSCRIPTION YOU WENT 4541 03:26:38,360 --> 03:26:41,760 GET EFFECT OF THAT RAPIDITY. 4542 03:26:41,760 --> 03:26:44,960 AND SO ON AVERAGE, THE RETINA, 4543 03:26:44,960 --> 03:26:49,640 PHYSICAL DISTANCE OF THE RETINA 4544 03:26:49,640 --> 03:26:50,840 SHRINKS AN AVERAGE OF 3.3 4545 03:26:50,840 --> 03:26:52,280 MICRONS IN THE PRESENCE OF 4546 03:26:52,280 --> 03:27:00,080 COBALT BUT THE MAJORITY OF THE 4547 03:27:00,080 --> 03:27:02,360 SHRINKAGE IS HERE, SIGNIFICANT 4548 03:27:02,360 --> 03:27:06,920 PORTION OF THE OUTER SEGMENTS OF 4549 03:27:06,920 --> 03:27:08,640 THE PHOTORECEPTORS ARE BEING 4550 03:27:08,640 --> 03:27:11,760 AFFECTED BY THE COBALT. 4551 03:27:11,760 --> 03:27:22,240 AND -- HOLD ON ONE MOMENT. 4552 03:27:26,200 --> 03:27:28,320 HERE YOU CAN SEE ON THE RIGHT 4553 03:27:28,320 --> 03:27:30,400 COMPARISON OF THREE RETINAS 4554 03:27:30,400 --> 03:27:32,160 WHERE WE APPLIED COBALT BEFORE 4555 03:27:32,160 --> 03:27:35,240 AND AFTER, AND YOU CAN SEE IN 4556 03:27:35,240 --> 03:27:37,640 EVERY CASE THESE LINES 4557 03:27:37,640 --> 03:27:39,040 REPRESENTING THE PHOTORECEPTOR 4558 03:27:39,040 --> 03:27:41,320 AREA SHOW THE ACTUAL 4559 03:27:41,320 --> 03:27:42,600 PHOTORECEPTORS ARE SHRIVELED 4560 03:27:42,600 --> 03:27:49,000 SLIGHTLY IN THE PRESENCE OF 4561 03:27:49,000 --> 03:27:49,240 COBALT. 4562 03:27:49,240 --> 03:27:50,320 OKAY. 4563 03:27:50,320 --> 03:27:51,640 EXCUSE ME. 4564 03:27:51,640 --> 03:27:54,160 SO, NOW WE CAN LOOK AT THE 4565 03:27:54,160 --> 03:27:58,440 COMPARISON OF THE EFFECTS OF 4566 03:27:58,440 --> 03:27:59,440 COBALT, ON CROSS-SECTIONAL 4567 03:27:59,440 --> 03:28:00,280 PROFILE. 4568 03:28:00,280 --> 03:28:01,960 YOU CAN SEE AGAIN THE BEFORE AND 4569 03:28:01,960 --> 03:28:03,280 AFTER THE COBALT, BEFORE IS 4570 03:28:03,280 --> 03:28:04,240 BLUE, AFTER IS RED. 4571 03:28:04,240 --> 03:28:06,960 AND YOU CAN SEE THERE'S A LARGE 4572 03:28:06,960 --> 03:28:10,360 DROP IN THIS PEAK HERE ON THE 4573 03:28:10,360 --> 03:28:11,240 INTERDIGITIZATION ZONE IN 4574 03:28:11,240 --> 03:28:13,320 PRESENCE OF COBALT AND ALSO PEAK 4575 03:28:13,320 --> 03:28:15,280 OF INNER/OUTER SEGMENT ZONE. 4576 03:28:15,280 --> 03:28:17,800 SO THE QUESTION THAT WE HAVE, 4577 03:28:17,800 --> 03:28:20,880 THIS SHOWS A CROSS-SECTION WITH 4578 03:28:20,880 --> 03:28:27,440 TIME, OF THE EFFECT ON THE 4579 03:28:27,440 --> 03:28:30,400 INNER/OUTER SEGMENT LINE, DROP 4580 03:28:30,400 --> 03:28:34,440 IN REFLEXIVITY WITH TIME, 4581 03:28:34,440 --> 03:28:36,280 DECLINED AVERAGE OF NINE UNITS, 4582 03:28:36,280 --> 03:28:38,280 SO PRETTY CLEAR THIS IS EFFECT 4583 03:28:38,280 --> 03:28:39,600 ON OUTER RETINA, AND THEN WE 4584 03:28:39,600 --> 03:28:41,760 WANT TO KNOW WHAT IS THE DOSE 4585 03:28:41,760 --> 03:28:43,640 AND WHAT RELEVANCE DOES IT HAVE 4586 03:28:43,640 --> 03:28:44,840 TO HUMAN VALUES? 4587 03:28:44,840 --> 03:28:46,920 WELL, A THOUSAND PARTS PER 4588 03:28:46,920 --> 03:28:49,080 MILLION, ABOUT THE WORST DOSE 4589 03:28:49,080 --> 03:28:53,040 REPORTED IN PATIENTS, WORKS OUT 4590 03:28:53,040 --> 03:28:54,760 TO 17 MICROMOLAR COBALT. 4591 03:28:54,760 --> 03:28:56,200 SO THAT'S -- THINK ABOUT THAT, 4592 03:28:56,200 --> 03:28:59,280 THAT'S A PATIENT THAT WOULD HAVE 4593 03:28:59,280 --> 03:29:01,000 COBALT THREE, FOUR YEARS IN 4594 03:29:01,000 --> 03:29:04,080 THEIR IMPLANT, SO IT'S A LONG 4595 03:29:04,080 --> 03:29:04,440 EXPOSURE TIME. 4596 03:29:04,440 --> 03:29:06,160 AND SO WHAT WE'VE DONE HERE IS 4597 03:29:06,160 --> 03:29:12,080 WE'VE STARTED TO DO DOSIMETRY, 4598 03:29:12,080 --> 03:29:15,800 YOU CAN GET THE SAME EFFECT OF 4599 03:29:15,800 --> 03:29:18,240 COBALT ON THE PHOTORECEPTOR 4600 03:29:18,240 --> 03:29:20,360 LINES AT 200 MICROMOLAR AND 4601 03:29:20,360 --> 03:29:24,440 SMALLER EFFECTS AT 40 4602 03:29:24,440 --> 03:29:27,280 MICROMOLAR, NOT FAR OFF FROM THE 4603 03:29:27,280 --> 03:29:29,280 DOSES THAT ARE AFFECTING 4604 03:29:29,280 --> 03:29:29,560 PATIENTS. 4605 03:29:29,560 --> 03:29:30,480 SO, THOUSAND DOLLAR QUESTION WE 4606 03:29:30,480 --> 03:29:33,560 WANT TO KNOW IS HOW IS COBALT IN 4607 03:29:33,560 --> 03:29:35,320 FACT AFFECTING THE RETINA? 4608 03:29:35,320 --> 03:29:42,240 WE'VE PREVIOUSLY LOOKED AT A 4609 03:29:42,240 --> 03:29:44,280 SERIES OF DIFFERENT 4610 03:29:44,280 --> 03:29:46,360 NEUROTOXICANTS, A DIP 8, NMDA, 4611 03:29:46,360 --> 03:29:50,720 AND WE'VE LOOKED AT CYANIDE AND 4612 03:29:50,720 --> 03:29:54,320 CHLOROQUIN, A WELL-KNOWN DRUG 4613 03:29:54,320 --> 03:29:55,440 THAT AFFECTS MITOCHONDRIA, AND 4614 03:29:55,440 --> 03:29:59,800 WHAT YOU CAN SEE HERE IS HERE WE 4615 03:29:59,800 --> 03:30:02,000 HAVE THE RETINA BEFORE AND AFTER 4616 03:30:02,000 --> 03:30:02,600 COBALT. 4617 03:30:02,600 --> 03:30:06,400 AND YOU CAN CLEARLY SEE THIS 4618 03:30:06,400 --> 03:30:07,480 CHANGE IN THE INTERDIGITIZATION 4619 03:30:07,480 --> 03:30:10,240 ZONE AND ALSO CHANGE IN THE 4620 03:30:10,240 --> 03:30:14,240 BRIGHTNESS OF THE INNER/OUTER 4621 03:30:14,240 --> 03:30:14,520 SEGMENTS. 4622 03:30:14,520 --> 03:30:15,440 SAME RABBIT RETINA, DIFFERENT 4623 03:30:15,440 --> 03:30:19,720 EXPERIMENT, IN THIS CASE USING 4624 03:30:19,720 --> 03:30:25,320 CYANIDE, A WELL-KNOWN METABOLIC 4625 03:30:25,320 --> 03:30:25,880 UNCOUPLER MUCH MITOCHONDRIA, 4626 03:30:25,880 --> 03:30:27,400 SAME LOSS NEXT TO THE RPE AND 4627 03:30:27,400 --> 03:30:30,120 ALSO SEE A LOSS IN THE 4628 03:30:30,120 --> 03:30:31,280 BRIGHTNESS OF THE INNER/OUTER 4629 03:30:31,280 --> 03:30:31,560 SEGMENTS. 4630 03:30:31,560 --> 03:30:34,320 ON THE RIGHT YOU CAN SEE 4631 03:30:34,320 --> 03:30:37,400 HISTOLOGY PICTURE OF BEFORE AND 4632 03:30:37,400 --> 03:30:38,760 AFTER CYANIDE SHOWING THIS 4633 03:30:38,760 --> 03:30:41,240 DISRUPTION OF THE PHOTORECEPTOR 4634 03:30:41,240 --> 03:30:41,440 TIPS. 4635 03:30:41,440 --> 03:30:48,520 WE CAN SEE DISRUPTION OF THE 4636 03:30:48,520 --> 03:30:49,480 PHOTORECEPTOR TIPS, THE THOUSAND 4637 03:30:49,480 --> 03:30:51,560 DOLLAR QUESTION WE'RE WORKING ON 4638 03:30:51,560 --> 03:30:53,960 CURRENTLY IS TRYING TO 4639 03:30:53,960 --> 03:30:56,040 UNDERSTAND HOW COBALT IS CAUSING 4640 03:30:56,040 --> 03:30:57,520 THIS EFFECT. 4641 03:30:57,520 --> 03:31:00,520 SO WE'VE DEVELOPED A METHOD TO 4642 03:31:00,520 --> 03:31:02,160 IMAGE METAL IONS RELEASED ON 4643 03:31:02,160 --> 03:31:04,040 RETINAL STRUCTURE IN REAL TIME, 4644 03:31:04,040 --> 03:31:06,760 SO THIS SAVES ON SACRIFICING 4645 03:31:06,760 --> 03:31:08,320 ANIMALS AT MULTIPLE TIME POINTS. 4646 03:31:08,320 --> 03:31:12,360 COBALT CAUSES A RAPID THINNING 4647 03:31:12,360 --> 03:31:14,560 IN THE RABBIT RETINA ON THE 4648 03:31:14,560 --> 03:31:18,160 ORDER OF MINUTES, NOT ACCOUNTED 4649 03:31:18,160 --> 03:31:25,680 FOR HYPOXIA FACTOR TYPE, GENE 4650 03:31:25,680 --> 03:31:27,680 ALTERATION, AM OPT POOK ARE POCK 4651 03:31:27,680 --> 03:31:34,040 ARE R. L. -- ALTERED POSITION OF 4652 03:31:34,040 --> 03:31:34,720 IS/OS LINE. 4653 03:31:34,720 --> 03:31:36,760 THERE ARE A FEW SMALLER EFFECTS 4654 03:31:36,760 --> 03:31:40,480 ON THE OUTER SIDE OF THE RETINA, 4655 03:31:40,480 --> 03:31:44,440 AFFECTING GANGLION CELLS, SO THE 4656 03:31:44,440 --> 03:31:46,320 RABBIT EYE OFFERS RETIME 4657 03:31:46,320 --> 03:31:47,520 ASSESSMENT OF TOXICANTS ON THE 4658 03:31:47,520 --> 03:31:48,480 RETINAL STRUCTURE AND WE'RE 4659 03:31:48,480 --> 03:31:51,320 GOING TO OFFER IT ACTUALLY AS A 4660 03:31:51,320 --> 03:31:52,080 FREE REGULATORY SCIENCE TOOL 4661 03:31:52,080 --> 03:31:53,680 THAT YOU CAN USE FOR STUDIES. 4662 03:31:53,680 --> 03:31:58,320 I DON'T KNOW IF YOU CAN DO 4663 03:31:58,320 --> 03:31:59,320 ANYTHING WITH SICKLE CELL WITH 4664 03:31:59,320 --> 03:32:00,520 IT BUT YOU'RE WELCOME TO IT. 4665 03:32:00,520 --> 03:32:02,600 THIS IS WHAT IT LOOKS LIKE. 4666 03:32:02,600 --> 03:32:03,920 HERE ARE MY COLLABORATORS, AND I 4667 03:32:03,920 --> 03:32:06,880 ALSO WORK WITH PEOPLE AT THE EYE 4668 03:32:06,880 --> 03:32:11,280 INSTITUTE AS WELL. 4669 03:32:11,280 --> 03:32:17,800 SO JOHANNA, STEVE, ROSLYN, THANK 4670 03:32:17,800 --> 03:32:18,000 YOU. 4671 03:32:18,000 --> 03:32:20,000 >> IN THE INTEREST OF TIME, 4672 03:32:20,000 --> 03:32:21,640 SAVE YOUR QUESTIONS FOR THE VERY 4673 03:32:21,640 --> 03:32:22,320 END IF POSSIBLE. 4674 03:32:22,320 --> 03:32:25,840 WE ALSO HAVE A POSTER 4675 03:32:25,840 --> 03:32:27,440 PRESENTATION FROM ALL THE SHORT 4676 03:32:27,440 --> 03:32:28,760 TALK SPEAKERS TOMORROW. 4677 03:32:28,760 --> 03:32:31,200 MAKE SURE YOU ATTEND THEIR 4678 03:32:31,200 --> 03:32:37,400 POSTER AND ASK ANY QUESTIONS YOU 4679 03:32:37,400 --> 03:32:37,720 MIGHT HAVE. 4680 03:32:37,720 --> 03:32:38,040 >> QUESTIONS? 4681 03:32:38,040 --> 03:32:44,000 >>WE'LL SAVE IT FOR LATER. 4682 03:32:44,000 --> 03:32:49,480 NEXT DR. QU TIAN, NATIONAL 4683 03:32:49,480 --> 03:32:50,080 INSTITUTE OF AGING. 4684 03:32:50,080 --> 03:32:51,440 >> GREAT TO BE HERE. 4685 03:32:51,440 --> 03:32:54,400 THANK YOU FOR THE ORGANIZERS OF 4686 03:32:54,400 --> 03:32:55,600 THE EVENT AND OPPORTUNITY TO 4687 03:32:55,600 --> 03:32:57,680 PRESENT SOME OF OUR WORK. 4688 03:32:57,680 --> 03:32:59,760 MY TITLE IS MITOCHONDRIAL 4689 03:32:59,760 --> 03:33:03,720 FUNCTION PREDICT IT'S COGNITIVE 4690 03:33:03,720 --> 03:33:11,040 IMPAIRMENT, ASSOCIATED WITH 4691 03:33:11,040 --> 03:33:17,600 BIOMARKERS IN NEURODEGENERATION. 4692 03:33:17,600 --> 03:33:18,520 SO, MITOCHONDRIA CASCADE 4693 03:33:18,520 --> 03:33:22,120 HYPOTHESIS WAS FIRST PROPOSED IN 4694 03:33:22,120 --> 03:33:23,400 2004, STATES MITOCHONDRIAL 4695 03:33:23,400 --> 03:33:24,320 FUNCTION WAS GENETICALLY 4696 03:33:24,320 --> 03:33:26,960 DETERMINED AT BIRTH. 4697 03:33:26,960 --> 03:33:28,400 OVER TIME, MITOCHONDRIA FUNCTION 4698 03:33:28,400 --> 03:33:31,080 WILL DECLINE WITH AGING. 4699 03:33:31,080 --> 03:33:33,720 ONCE THE DECLINE PASSED CERTAIN 4700 03:33:33,720 --> 03:33:37,120 THRESHOLD IMAGE MAY TRIGGER 4701 03:33:37,120 --> 03:33:46,080 FUTURE ALZHEIMER'S PATHOLOGY. 4702 03:33:46,080 --> 03:33:50,360 SO OVER THE YEARS THERE HAS BEEN 4703 03:33:50,360 --> 03:33:51,000 RESEARCH DEMONSTRATING 4704 03:33:51,000 --> 03:33:52,360 MITOCHONDRIA DYSFUNCTION IS 4705 03:33:52,360 --> 03:33:53,640 IMPORTANT HALLMARK OF AGING. 4706 03:33:53,640 --> 03:33:56,960 AND THERE HAS ALSO BEEN EVIDENCE 4707 03:33:56,960 --> 03:34:02,440 SHOWING CONNECTION BETWEEN 4708 03:34:02,440 --> 03:34:06,360 MITOCHONDRIA DYSFUNCTION WITH 4709 03:34:06,360 --> 03:34:11,040 A-BETA AND TAU, HALLMARKS OF 4710 03:34:11,040 --> 03:34:14,840 ALZHEIMER'S DISEASE, MOST OF THE 4711 03:34:14,840 --> 03:34:16,200 EVIDENCE COMING FROM ANIMAL 4712 03:34:16,200 --> 03:34:19,280 MODELS, THERE'S BEEN LIMITED 4713 03:34:19,280 --> 03:34:21,040 EVIDENCE FROM HUMAN STUDIES. 4714 03:34:21,040 --> 03:34:23,640 AND ALONG THE LINE WE 4715 03:34:23,640 --> 03:34:24,560 HYPOTHESIZE MITOCHONDRIAL 4716 03:34:24,560 --> 03:34:27,360 DYSFUNCTION WOULD CONTRIBUTE TO 4717 03:34:27,360 --> 03:34:30,080 FUTURE DECLINE AND ALSO 4718 03:34:30,080 --> 03:34:31,600 IMPAIRMENT OF COGNITION, 4719 03:34:31,600 --> 03:34:34,680 POTENTIALLY THROUGH LOW ENERGY 4720 03:34:34,680 --> 03:34:37,240 AVAILABILITY, INCREASED STRESS 4721 03:34:37,240 --> 03:34:38,080 AND INFLAMMATION. 4722 03:34:38,080 --> 03:34:39,840 WE'RE USING DATA FROM THE 4723 03:34:39,840 --> 03:34:41,600 BALTIMORE LONGITUDINAL STUDY OF 4724 03:34:41,600 --> 03:34:45,720 AGING WHICH IS THE AMERICAN'S 4725 03:34:45,720 --> 03:34:48,600 LONGEST RUNNING STUDY, BEGAN IN 4726 03:34:48,600 --> 03:34:50,120 1958, PARTICIPANTS COMING FROM 4727 03:34:50,120 --> 03:34:56,160 THE COMMUNITY, SO WE'RE TESTING 4728 03:34:56,160 --> 03:34:57,680 THE HYPOTHESIS WHETHER 4729 03:34:57,680 --> 03:34:59,240 MITOCHONDRIAL FUNCTION WILL 4730 03:34:59,240 --> 03:35:00,120 PREDICT FUTURE DEVELOPMENT OF 4731 03:35:00,120 --> 03:35:01,720 COGNITIVE IMPAIRMENT OR 4732 03:35:01,720 --> 03:35:03,800 DEMENTIA. 4733 03:35:03,800 --> 03:35:07,000 WE IDENTIFIED 469 PARTICIPANTS, 4734 03:35:07,000 --> 03:35:08,400 INITIALLY COGNITIVE NORMAL, 4735 03:35:08,400 --> 03:35:11,040 AVERAGE AGE 70, DIVERSE SAMPLE 4736 03:35:11,040 --> 03:35:14,720 WITH MANY ETHNICITIES AND RACE, 4737 03:35:14,720 --> 03:35:19,320 AND FOLLOW-UP FIVE YEARS, 5% 4738 03:35:19,320 --> 03:35:20,760 MITOCHONDRIAL IMPAIRMENT OR 4739 03:35:20,760 --> 03:35:21,360 DEMENTIA. 4740 03:35:21,360 --> 03:35:23,920 IN SUBSET WE ALSO LOOK AT 4741 03:35:23,920 --> 03:35:26,680 ASSOCIATIONS BETWEEN IN VIVO 4742 03:35:26,680 --> 03:35:28,320 MITOCHONDRIAL FUNCTION WITH 4743 03:35:28,320 --> 03:35:36,120 BIOMARKERS ALZHEIMER'S DISEASE 4744 03:35:36,120 --> 03:35:38,200 OF THE BRAIN. 4745 03:35:38,200 --> 03:35:43,120 THE SPECIFIC MEASURE HERE WITH 4746 03:35:43,120 --> 03:35:48,120 THE SKELETAL FUNCTION IS MAXIMAL 4747 03:35:48,120 --> 03:35:49,760 CAPACITY, IN VIVO ASSESSMENT. 4748 03:35:49,760 --> 03:35:53,080 YOU SEE THE PART PANTS ARE IN 4749 03:35:53,080 --> 03:35:56,320 THE SCANNER, AND THEY ARE 4750 03:35:56,320 --> 03:35:57,320 INSTRUCTED TO PERFORM INTENSE 4751 03:35:57,320 --> 03:35:59,960 EXERCISE UNTIL A POINT OF 4752 03:35:59,960 --> 03:36:06,640 EXHAUSTION, AND THE GOAL OF THE 4753 03:36:06,640 --> 03:36:08,080 EXERCISE PROTOCOL TO DEPLETE 4754 03:36:08,080 --> 03:36:11,240 PHOSPHO-CREATINE IN THE MUSCLES. 4755 03:36:11,240 --> 03:36:16,920 SOLID LINE INDICATES CHANGE IN 4756 03:36:16,920 --> 03:36:17,240 THE MUSCLE. 4757 03:36:17,240 --> 03:36:19,440 WE'RE CAPTURING RECOVERY RATE IN 4758 03:36:19,440 --> 03:36:20,440 THE SKELETAL MUSCLE. 4759 03:36:20,440 --> 03:36:22,520 SO THE RATE OF RECOVERY 4760 03:36:22,520 --> 03:36:25,240 INDICATES A HIGHER -- FASTER 4761 03:36:25,240 --> 03:36:27,080 RECOVERY RATE INDICATES HIGHER 4762 03:36:27,080 --> 03:36:28,720 OXIDATIVE CAPACITY AND HIGHER 4763 03:36:28,720 --> 03:36:33,320 MITOCHONDRIAL FUNCTION. 4764 03:36:33,320 --> 03:36:36,640 HERE WE DENOTE AS KPCR. 4765 03:36:36,640 --> 03:36:39,040 WE'VE FOUND THIS HAS BEEN 4766 03:36:39,040 --> 03:36:43,320 ASSOCIATED WITH EX VIVO 4767 03:36:43,320 --> 03:36:53,840 MITOCHONDRIA RESPIRATION FROM 4768 03:37:01,680 --> 03:37:02,480 MUSCLE BIOPSY. 4769 03:37:02,480 --> 03:37:03,480 ACROSS TISSUES WE'VE SEEN 4770 03:37:03,480 --> 03:37:05,880 CORRELATION BETWEEN IN VIVO 4771 03:37:05,880 --> 03:37:07,600 OXIDATIVE CAPACITY WITH 4772 03:37:07,600 --> 03:37:08,840 DIFFERENT MITOCHONDRIA MARKERS. 4773 03:37:08,840 --> 03:37:11,680 AND FROM THE STUDY WE FOUND THAT 4774 03:37:11,680 --> 03:37:14,440 THE PARTICIPANT HAD HIGHER 4775 03:37:14,440 --> 03:37:16,280 MITOCHONDRIAL FUNCTION AT 4776 03:37:16,280 --> 03:37:19,600 BASELINE, THEY HAD A HIGHER 4777 03:37:19,600 --> 03:37:21,560 PROBABILITY OF BEING COGNITIVE 4778 03:37:21,560 --> 03:37:23,840 NORMAL, LOWER RISK OF DEVELOPING 4779 03:37:23,840 --> 03:37:26,440 COGNITIVE IMPAIRMENT OR DEMENTIA 4780 03:37:26,440 --> 03:37:28,320 COMPARED TO LOW MITOCHONDRIAL 4781 03:37:28,320 --> 03:37:29,400 FUNCTION AT BASELINE, 4782 03:37:29,400 --> 03:37:31,280 INDEPENDENT OF THE DEMOGRAPHIC 4783 03:37:31,280 --> 03:37:38,480 FACTORS AS WELL AS APOE 4 4784 03:37:38,480 --> 03:37:39,800 CARRIER STATUS. 4785 03:37:39,800 --> 03:37:44,760 WE FOUND THIS IS ASSOCIATED WITH 4786 03:37:44,760 --> 03:37:46,720 BIOMARKERS OF ALZHEIMER'S 4787 03:37:46,720 --> 03:37:50,480 DISEASE, HERE AMYLOID TRACER 4788 03:37:50,480 --> 03:37:51,840 UPTAKE BASED ON PET IMAGING, 4789 03:37:51,840 --> 03:37:52,960 ASSOCIATION PRESENT IN GRAY 4790 03:37:52,960 --> 03:37:54,360 MATTER AND WHITE MATTER. 4791 03:37:54,360 --> 03:37:56,800 AND WE ALSO FOUND THERE'S 4792 03:37:56,800 --> 03:38:00,040 ASSOCIATION WITH BIOMARKERS OF 4793 03:38:00,040 --> 03:38:03,280 NEURODEGENERATION INDICATED BY 4794 03:38:03,280 --> 03:38:07,920 THE GFAP, AND ALSO A TREND 4795 03:38:07,920 --> 03:38:17,560 TOWARDS LOWER NfL, INDICATOR 4796 03:38:17,560 --> 03:38:19,880 OF NEURONAL INJURY. 4797 03:38:19,880 --> 03:38:22,040 WE FOUND THE STRONGEST 4798 03:38:22,040 --> 03:38:25,320 ASSOCIATION HAS BEEN LOCALIZED 4799 03:38:25,320 --> 03:38:28,840 TO SPECIFIC BRAIN AREA INCLUDING 4800 03:38:28,840 --> 03:38:35,920 PRE-CENTRAL GYRUS, POST CENTRAL 4801 03:38:35,920 --> 03:38:37,640 GYRUS AND LOBE, IMPORTANT FOR 4802 03:38:37,640 --> 03:38:41,000 MOTOR FUNCTION AND EXECUTION. 4803 03:38:41,000 --> 03:38:41,440 ASSOCIATION BETWEEN 4804 03:38:41,440 --> 03:38:43,720 MITOCHONDRIAL FUNCTION AND 4805 03:38:43,720 --> 03:38:47,000 AMYLOID TRACER UPTAKE HAS BEEN 4806 03:38:47,000 --> 03:38:50,280 SPECIFIC TO SPECIFIC REGIONS. 4807 03:38:50,280 --> 03:38:53,560 IN SUMMARY THIS FINDING PROVIDES 4808 03:38:53,560 --> 03:38:56,760 THE FIRST EVIDENCE OF IN VIVO 4809 03:38:56,760 --> 03:38:58,800 MITOCHONDRIAL FUNCTION TO THE 4810 03:38:58,800 --> 03:39:00,000 FUTURE COGNITIVE IMPAIRMENT AND 4811 03:39:00,000 --> 03:39:03,840 FUNDING FROM THE BELL MARKERS 4812 03:39:03,840 --> 03:39:07,480 MAY INDICATE MITOCHONDRIAL 4813 03:39:07,480 --> 03:39:10,240 DYSFUNCTION MAY PLAY KEY ROLE IN 4814 03:39:10,240 --> 03:39:11,520 A.D. POTENTIAL 4815 03:39:11,520 --> 03:39:14,120 APPROACHESOLOGICAL CHANGES AND 4816 03:39:14,120 --> 03:39:14,680 NEUROINFLAMMATION. 4817 03:39:14,680 --> 03:39:24,240 FUTURE STUDIES SHOULD FOCUS ON 4818 03:39:24,240 --> 03:39:24,760 LONGITUDINAL EXAMINATION 4819 03:39:24,760 --> 03:39:26,320 ALSO INVESTIGATE OTHER IMPORTANT 4820 03:39:26,320 --> 03:39:27,600 MECHANISMS SUCH AS MITOCHONDRIAL 4821 03:39:27,600 --> 03:39:29,800 FUNCTION IN THE BRAIN WHICH CAN 4822 03:39:29,800 --> 03:39:34,000 BE ASSESSED BY M.R. SPECTROSCOPY 4823 03:39:34,000 --> 03:39:35,960 AS WELL AND PERFUSION, IRON 4824 03:39:35,960 --> 03:39:38,360 DEPRIVATION, ALSO ONE OF THE 4825 03:39:38,360 --> 03:39:40,320 IMPORTANT MECHANISMS OF 4826 03:39:40,320 --> 03:39:41,960 OXIDATIVE STRESS, ALSO WHITE 4827 03:39:41,960 --> 03:39:43,720 MATTER HEALTH IN THE BRAIN. 4828 03:39:43,720 --> 03:39:46,800 BASED ON THE OBSERVATIONAL 4829 03:39:46,800 --> 03:39:48,960 STUDIES IT'S IMPORTANT TO NOTE 4830 03:39:48,960 --> 03:39:50,840 THERE'S BEEN PRELIMINARY 4831 03:39:50,840 --> 03:39:52,040 EVIDENCE FROM THE LITERATURE 4832 03:39:52,040 --> 03:39:53,920 THAT MITOCHONDRIAL FUNCTION CAN 4833 03:39:53,920 --> 03:39:58,480 BE IMPROVED IN HUMANS THROUGH 4834 03:39:58,480 --> 03:40:00,240 BOTH NON-PHARMACOLOGICAL 4835 03:40:00,240 --> 03:40:03,080 APPROACH AND SUPPLEMENTS, AND 4836 03:40:03,080 --> 03:40:05,280 THERE'S SOME EVIDENCE FROM 4837 03:40:05,280 --> 03:40:07,120 TRIGLYCERIDES, SO OUR STUDIES 4838 03:40:07,120 --> 03:40:09,960 LAY A FOUNDATION FOR FUTURE EYE 4839 03:40:09,960 --> 03:40:10,640 INTERVENTION STUDIES, 4840 03:40:10,640 --> 03:40:12,600 EXPERIMENTAL STUDY TO TEST 4841 03:40:12,600 --> 03:40:13,480 WHETHER IMPROVING MITOCHONDRIAL 4842 03:40:13,480 --> 03:40:16,440 FUNCTION MAY IMPROVE BRAIN 4843 03:40:16,440 --> 03:40:21,240 HEALTH OR MUSCLE FUNCTION WITHIN 4844 03:40:21,240 --> 03:40:21,440 AGING. 4845 03:40:21,440 --> 03:40:23,440 SO WITH THAT, I'D LIKE TO 4846 03:40:23,440 --> 03:40:25,720 ACKNOWLEDGE THE STUDY 4847 03:40:25,720 --> 03:40:28,560 PARTICIPANTS AND STAFF AND MANY 4848 03:40:28,560 --> 03:40:29,640 CO-AUTHORS IN THIS WORK. 4849 03:40:29,640 --> 03:40:31,080 AND IN THE INTEREST OF TIME I'M 4850 03:40:31,080 --> 03:40:33,600 HAPPY TO TAKE ANY QUESTIONS. 4851 03:40:33,600 --> 03:40:35,040 WELL, WE'LL TAKE QUESTIONS 4852 03:40:35,040 --> 03:40:36,480 TOMORROW. 4853 03:40:36,480 --> 03:40:37,080 ALL RIGHT, THANK YOU. 4854 03:40:37,080 --> 03:40:39,080 [APPLAUSE] 4855 03:40:39,080 --> 03:40:43,240 4856 03:40:43,240 --> 03:40:44,560 4857 03:40:44,560 --> 03:40:55,120 >>LAST SPEAKER IS A A BIOMEDIL 4858 03:40:56,680 --> 03:40:56,920 UPPING NEAR. 4859 03:40:56,920 --> 03:41:00,000 TODAY I WOULD LIKE TO TALK TO 4860 03:41:00,000 --> 03:41:05,960 YOU ABOUT MARKER FABRICATION AND 4861 03:41:05,960 --> 03:41:08,000 MICROFLUIDICS FOR CELL BIOLOGY. 4862 03:41:08,000 --> 03:41:13,000 BACKGROUND, I WORK AT 4863 03:41:13,000 --> 03:41:18,480 MICROFABRICATION AND MICROFLUID 4864 03:41:18,480 --> 03:41:20,400 UNITS, UNDER BIOMEDICAL 4865 03:41:20,400 --> 03:41:22,040 ENGINEERING, NIBIB, IN BUILDING 4866 03:41:22,040 --> 03:41:22,560 13. 4867 03:41:22,560 --> 03:41:26,080 SO, LET ME START WITH 4868 03:41:26,080 --> 03:41:30,560 REINTRODUCING YOU TO MARKER 4869 03:41:30,560 --> 03:41:31,240 FABRICATION, DEVELOPED FOR 4870 03:41:31,240 --> 03:41:32,640 SEMICONDUCTOR INDUSTRY, CAPABLE 4871 03:41:32,640 --> 03:41:34,840 OF PATTERNING FEATURES DOWN TO 4872 03:41:34,840 --> 03:41:36,240 ONE MICRONS. 4873 03:41:36,240 --> 03:41:38,120 NOW, WHEN YOU GET TO MANIPULATE 4874 03:41:38,120 --> 03:41:41,960 FLUIDS AND GASES IN CHANNELS 4875 03:41:41,960 --> 03:41:46,640 THAT ARE IN MICROMETER RANGE 4876 03:41:46,640 --> 03:41:47,320 THAT'S CALLED MICROFLUIDICS, 4877 03:41:47,320 --> 03:41:49,840 USED AND APPLIED IN MANY FIELDS. 4878 03:41:49,840 --> 03:41:51,760 NOW, THE REASON IT IS IMPORTANT 4879 03:41:51,760 --> 03:41:56,480 FOR US IS BECAUSE WE CAN USE 4880 03:41:56,480 --> 03:41:57,960 MICROFLUIDICS AND 4881 03:41:57,960 --> 03:42:00,040 MICROFABRICATION TO REPLICATE 4882 03:42:00,040 --> 03:42:04,000 CELLULAR MICROENVIRONMENT AND 4883 03:42:04,000 --> 03:42:05,200 CREATE SMALL SCALE SYSTEMS. 4884 03:42:05,200 --> 03:42:07,480 IN THE NEXT FEW SLIDES I'LL TALK 4885 03:42:07,480 --> 03:42:09,760 ABOUT ONE OF THE PROJECTS WE 4886 03:42:09,760 --> 03:42:15,920 HAVE IN COLLABORATION WITH 4887 03:42:15,920 --> 03:42:18,760 RESEARCHERS, THEY WANTED TO 4888 03:42:18,760 --> 03:42:21,040 DEFINE NEUTROPHIL HETEROGENEITY 4889 03:42:21,040 --> 03:42:22,920 IN PATIENTS WITH SLE, BUT 4890 03:42:22,920 --> 03:42:24,280 UNDERSTANDING THE EFFECTS OF 4891 03:42:24,280 --> 03:42:25,520 MECHANICAL PRODUCTS OF 4892 03:42:25,520 --> 03:42:29,040 NEUTROPHILS AS THEY ARE 4893 03:42:29,040 --> 03:42:29,680 TRAFFICKING THROUGH PULMONARY 4894 03:42:29,680 --> 03:42:35,640 MICRO VATS -- MICROVASCULATURE, 4895 03:42:35,640 --> 03:42:37,720 COMPARING ROUGHNESS OF NORMAL 4896 03:42:37,720 --> 03:42:41,320 AND LOW DENSITY NEUTROPHILS IN 4897 03:42:41,320 --> 03:42:43,200 SLE PATIENTS WITH HEALTHY 4898 03:42:43,200 --> 03:42:44,080 CONTROLLED NORMAL DENSITY 4899 03:42:44,080 --> 03:42:48,440 NEUTROPHILS AND ONES THAT HAVE 4900 03:42:48,440 --> 03:42:49,440 THEM CHEMICALLY PRIMED. 4901 03:42:49,440 --> 03:42:51,560 THEY CAME WITH A ROUGH IDEA 4902 03:42:51,560 --> 03:42:53,200 ABOUT WHAT THEY WANTED TO DO. 4903 03:42:53,200 --> 03:42:58,480 AND HOW THEY WANTED TO QUANTIFY 4904 03:42:58,480 --> 03:42:59,800 THE NEUTROPHIL ROUGHNESS, AND 4905 03:42:59,800 --> 03:43:01,840 CORRELATE THAT WITH THE TIME IT 4906 03:43:01,840 --> 03:43:10,520 TAKES FOR THE CELL TO TRANSIT 4907 03:43:10,520 --> 03:43:11,960 THROUGH PULMONARY 4908 03:43:11,960 --> 03:43:12,920 MICROVASCULATURE, WE HAD 4909 03:43:12,920 --> 03:43:16,640 DISCUSSIONS, WE TOOK INTO 4910 03:43:16,640 --> 03:43:17,760 ACCOUNT THE READOUT REQUIREMENTS 4911 03:43:17,760 --> 03:43:19,400 AND CONSTRAINTS THEY HAD FOR THE 4912 03:43:19,400 --> 03:43:22,880 PROJECT, AND WE CAME UP WITH TWO 4913 03:43:22,880 --> 03:43:25,600 DIFFERENT PATTERNS, TWO 4914 03:43:25,600 --> 03:43:27,240 DIFFERENT DEVICE PATTERNS. 4915 03:43:27,240 --> 03:43:33,960 DESIGN 1, BRANCHING NETWORK 4916 03:43:33,960 --> 03:43:35,080 WHICH REPLICATES PULMONARY 4917 03:43:35,080 --> 03:43:36,880 VASCULATURE, DESIGN 2 MULTIPLE 4918 03:43:36,880 --> 03:43:39,680 CONSTRUCTIONS, SINGLE CHANNEL. 4919 03:43:39,680 --> 03:43:41,400 THAT MIMICS CAPILLARY BED. 4920 03:43:41,400 --> 03:43:47,280 USING THE CAD DRAWINGS WE 4921 03:43:47,280 --> 03:43:47,920 ORDERED A PHOTOMASK. 4922 03:43:47,920 --> 03:43:51,360 ONCE WE HAVE THE MASK WE USE IT 4923 03:43:51,360 --> 03:43:56,160 IN A PROCESS CALLED 4924 03:43:56,160 --> 03:43:56,960 PHOTOLITERATUROGRAPHY, WE COAT 4925 03:43:56,960 --> 03:43:59,040 THE SURFACE WITH A 4926 03:43:59,040 --> 03:44:01,640 PHOTORESISTANT MATERIAL, SHINE 4927 03:44:01,640 --> 03:44:03,520 UV LIGHT THROUGH THE PHOTO MASK, 4928 03:44:03,520 --> 03:44:07,440 IN THIS CASE AREAS OF 4929 03:44:07,440 --> 03:44:08,240 PHOTORESIST, WILL CROSS-LINK, 4930 03:44:08,240 --> 03:44:09,880 EVERYTHING ELSE IS WASHED OFF. 4931 03:44:09,880 --> 03:44:13,560 WHAT YOU END UP WITH IS SILICONE 4932 03:44:13,560 --> 03:44:15,520 VAPOR WITH FEATURES THAT CAN BE 4933 03:44:15,520 --> 03:44:19,480 USEDDED A TEMPLATE OR MOLD TO 4934 03:44:19,480 --> 03:44:20,800 MAKE DEVICES. 4935 03:44:20,800 --> 03:44:25,160 ONCE YOU HAVE THOSE TEMPLATES, 4936 03:44:25,160 --> 03:44:27,840 WE CAST CURE IT, UNMOLD, CUT 4937 03:44:27,840 --> 03:44:29,160 THEM, PUNCH IN OUTLETS AND 4938 03:44:29,160 --> 03:44:36,360 PREPARE THEM FOR THE 4939 03:44:36,360 --> 03:44:36,680 EXPERIMENTS. 4940 03:44:36,680 --> 03:44:39,360 OUR COLLABORATORS TRIED ALL THE 4941 03:44:39,360 --> 03:44:40,440 DIFFERENT DESIGN ITERATIONS WE 4942 03:44:40,440 --> 03:44:42,840 PROVIDED TO THEM AND THEY ENDED 4943 03:44:42,840 --> 03:44:45,800 UP USING THESE BRANCHING 4944 03:44:45,800 --> 03:44:47,000 NETWORKS FOR THEIR EXPERIMENTS. 4945 03:44:47,000 --> 03:44:49,800 AND WHAT THEY DID WAS THAT THEY 4946 03:44:49,800 --> 03:44:51,920 INTRODUCED ONE CELL AT A TIME, 4947 03:44:51,920 --> 03:44:53,400 TO THIS BRANCHING NETWORK, AND 4948 03:44:53,400 --> 03:44:54,960 THEY MEASURED HOW LONG IT WILL 4949 03:44:54,960 --> 03:44:59,560 TAKE FOR A CELL TO ENTER AND 4950 03:44:59,560 --> 03:45:00,880 THEN EXIT. 4951 03:45:00,880 --> 03:45:03,240 THERE'S A CORRELATION BETWEEN 4952 03:45:03,240 --> 03:45:05,240 CELL ROUGHNESS AND HOW LONG IT 4953 03:45:05,240 --> 03:45:07,880 WILL -- HOW LONG THE CELLS WILL 4954 03:45:07,880 --> 03:45:11,520 STAY IN THE BRANCHING SYSTEM. 4955 03:45:11,520 --> 03:45:13,040 TO CHANGE GEARS A LITTLE BIT I'M 4956 03:45:13,040 --> 03:45:23,160 GOING TO 4957 03:45:27,320 --> 03:45:30,200 BEADS AND CAPSULES, HERE ON THE 4958 03:45:30,200 --> 03:45:32,600 LEFT YOU'RE LOOKING TO PHASES 4959 03:45:32,600 --> 03:45:36,960 COMING TOGETHER MAKING DROPS IN 4960 03:45:36,960 --> 03:45:43,000 OIL, USUALLY FOR EXPERIMENTS 4961 03:45:43,000 --> 03:45:44,960 LIKE THIS WE RUN THE CELLS LIEU, 4962 03:45:44,960 --> 03:45:47,680 EACH DROP WILL HAVE MAXIMUM OF 4963 03:45:47,680 --> 03:45:58,040 ONE CELL, ONE OF THE 4964 03:46:10,160 --> 03:46:13,560 METHODS FOR SORTING IS FLOW 4965 03:46:13,560 --> 03:46:15,200 CYTOMETRY BUT THE DROPS AT THE 4966 03:46:15,200 --> 03:46:19,240 STAIN ARE NOT COMPATIBLE WITH 4967 03:46:19,240 --> 03:46:20,360 FLOW CYTOMETRY BECAUSE THEY ARE 4968 03:46:20,360 --> 03:46:21,480 SURROUNDED BY OIL. 4969 03:46:21,480 --> 03:46:28,600 THERE ARE A ANOTHER SET OF 4970 03:46:28,600 --> 03:46:34,560 DEVICES, YOU'RE LOOKING AT HALF 4971 03:46:34,560 --> 03:46:36,320 HERE, IN THE FIRST PART OF THE 4972 03:46:36,320 --> 03:46:46,880 DEVICE WE HAVE THE WATER AND OIL 4973 03:46:55,280 --> 03:46:59,640 DROPS, AND ON THE SAME DEVICE 4974 03:46:59,640 --> 03:47:01,680 THE DROPS ARE REENCAPSULATED, 4975 03:47:01,680 --> 03:47:03,640 NOW WE HAVE WATER AND OIL IN 4976 03:47:03,640 --> 03:47:11,880 WATER DROPS THAT ARE NOW 4977 03:47:11,880 --> 03:47:12,760 COMPATIBLE WITH FLOW CYTOMETRY. 4978 03:47:12,760 --> 03:47:14,680 THE SKY IS THE LIMIT WITH 4979 03:47:14,680 --> 03:47:16,280 FABRICATION, I HAVE MORE 4980 03:47:16,280 --> 03:47:18,200 EXAMPLES FOR YOU. 4981 03:47:18,200 --> 03:47:21,080 HERE YOU'RE SEEING AN EXAMPLE OF 4982 03:47:21,080 --> 03:47:22,880 DEVICES USED FOR CULTURE AND 4983 03:47:22,880 --> 03:47:25,440 ISOLATION OF AXONS. 4984 03:47:25,440 --> 03:47:36,000 IN THE MIDDLE WE HAVE AN EXAMPLE 4985 03:47:36,720 --> 03:47:41,880 OF TISSUE, ON A CHIP, HERE 4986 03:47:41,880 --> 03:47:47,880 FIBROSIS ON A CHIP IS STUDIED. 4987 03:47:47,880 --> 03:47:52,680 LASTLY USE OF PBMS DEVICES FOR 4988 03:47:52,680 --> 03:47:56,000 PATTERNING SURFACES, YOU CAN SEE 4989 03:47:56,000 --> 03:48:01,560 PILLARS ABOUT ONE MILLIMETER IN 4990 03:48:01,560 --> 03:48:04,520 DIAMETER, COATED WITH MATRIGEL, 4991 03:48:04,520 --> 03:48:07,200 STAMPED ON A GLASS, CELLS ARE 4992 03:48:07,200 --> 03:48:14,720 SEATED THERE, YOU CAN SEE 4993 03:48:14,720 --> 03:48:15,160 SPATIAL ORGANIZATIONS. 4994 03:48:15,160 --> 03:48:16,320 I'LL BE AVAILABLE TOMORROW IF 4995 03:48:16,320 --> 03:48:18,560 YOU HAVE QUESTIONS ON THE POSTER 4996 03:48:18,560 --> 03:48:18,760 SESSION. 4997 03:48:18,760 --> 03:48:22,040 YOU CAN GO INTO THE WEBSITE AND 4998 03:48:22,040 --> 03:48:26,120 SEE WHAT CAPABILITIES WE HAVE ON 4999 03:48:26,120 --> 03:48:28,840 CAMPUS. 5000 03:48:28,840 --> 03:48:31,360 LASTLY, BUT NOT LEAST, I THANK 5001 03:48:31,360 --> 03:48:35,840 MY MENTOR DR. MORGAN, THE 5002 03:48:35,840 --> 03:48:36,840 ORGANIZERS, FOR THIS SEMINAR, 5003 03:48:36,840 --> 03:48:41,240 AND GIVING ME THE OPPORTUNITY TO 5004 03:48:41,240 --> 03:48:50,560 TALK AND THANK YOU ALL 5005 03:48:50,560 --> 03:48:50,880 EVERYTHING. 5006 03:48:50,880 --> 03:48:52,480 >>I HAVE A BUNCH OF QUESTIONS. 5007 03:48:52,480 --> 03:48:56,200 I HAVE TO CLOSE THE MEETING FOR 5008 03:48:56,200 --> 03:48:56,840 RECORDING PURPOSES. 5009 03:48:56,840 --> 00:00:00,000 THANK YOU SO MUCH, EVERYONE.