1 00:00:05,360 --> 00:00:09,680 >>WELCOME TO THE DNA REPAIR 2 00:00:09,680 --> 00:00:12,120 INTEREST GROUP VIDEO CONFERENCE. 3 00:00:12,120 --> 00:00:14,920 WE HAD A BREAK DURING THE SUMMER 4 00:00:14,920 --> 00:00:18,480 IN JULY AND AUGUST AND WE ARE 5 00:00:18,480 --> 00:00:21,200 STARTING AGAIN NOW IN SEPTEMBER. 6 00:00:21,200 --> 00:00:23,560 WITH THREE SPEAKERS. 7 00:00:23,560 --> 00:00:29,680 WE HAVE ARRANGED FOR SPEAKERS 8 00:00:29,680 --> 00:00:33,560 FOR OCTOBER 18TH. 9 00:00:33,560 --> 00:00:34,640 DR. (INAUDIBLE) FROM FRANCE WILL 10 00:00:34,640 --> 00:00:37,560 BE SPEAKING ON CHROMOSOME AND 11 00:00:37,560 --> 00:00:41,520 CHROMATIN DYNAMICS AND THEN ON 12 00:00:41,520 --> 00:00:43,960 NOVEMBER 15TH DR. (INAUDIBLE) 13 00:00:43,960 --> 00:00:45,560 ALSO FROM FRANCE WILL BE 14 00:00:45,560 --> 00:00:47,160 SPEAKING ABOUT TRANSCRIPTION AND 15 00:00:47,160 --> 00:00:49,640 DNA REPAIR. 16 00:00:49,640 --> 00:00:55,000 I WILL BE ENDING THE YEAR 17 00:00:55,000 --> 00:00:57,200 TALKING ABOUT ZERO PIGMENTOSA 18 00:00:57,200 --> 00:00:59,240 WHEN THE GENE POOL GOES ON 19 00:00:59,240 --> 00:01:02,000 STRIKE. 20 00:01:02,000 --> 00:01:04,000 >> I WANTED TO SAY WE ARE VERY 21 00:01:04,000 --> 00:01:06,720 PLEASED THAT THE VIDEO SERIES 22 00:01:06,720 --> 00:01:11,400 WILL CONTINUE AFTER NEXT YEAR, 23 00:01:11,400 --> 00:01:14,600 WITH JONG YANG AND KAREN AUSTIN 24 00:01:14,600 --> 00:01:16,240 WILL STEP IN AS ORGANIZERS OF 25 00:01:16,240 --> 00:01:21,080 THIS FOLLOWING THE MANY YEARS I 26 00:01:21,080 --> 00:01:23,880 HAVE BEEN DOING THIS, PLEASED TO 27 00:01:23,880 --> 00:01:26,160 HAVE CONTINUATION. 28 00:01:26,160 --> 00:01:27,360 JONG WILL BE PART OF THE TEAM 29 00:01:27,360 --> 00:01:28,600 TODAY AND TAKE THE QUESTIONS. 30 00:01:28,600 --> 00:01:31,080 COMING UP AFTER THE TALKS. 31 00:01:31,080 --> 00:01:32,720 SO THE ORGANIZATION IS THAT WE 32 00:01:32,720 --> 00:01:34,440 HAVE THE THREE SHORT TALKS THEN 33 00:01:34,440 --> 00:01:37,120 THE QUESTIONS WILL BE AFTER THE 34 00:01:37,120 --> 00:01:38,880 TALKS, QUESTIONS TO ALL THREE 35 00:01:38,880 --> 00:01:41,200 TALKS, THEY WILL COME THROUGH 36 00:01:41,200 --> 00:01:43,400 THE CHAT TO THE PANELISTS AND 37 00:01:43,400 --> 00:01:47,800 THEN READ THEM UP FOR THE 38 00:01:47,800 --> 00:01:48,400 SPEAKERS. 39 00:01:48,400 --> 00:01:50,240 ANYTHING ELSE, KEN? 40 00:01:50,240 --> 00:01:51,240 >> NO. 41 00:01:51,240 --> 00:01:56,760 YOU CAN DO THE INTRODUCTIONS. 42 00:01:56,760 --> 00:02:01,360 >> THE FIRST SPEAKER IS JONG 43 00:02:01,360 --> 00:02:03,840 HYUK LEE, I KNOW I DIDN'T 44 00:02:03,840 --> 00:02:05,840 PRONOUNCE IT PERFECTLY BUT JONG 45 00:02:05,840 --> 00:02:09,520 COMES FROM SOUTH KOREA WHERE HE 46 00:02:09,520 --> 00:02:11,760 WAS A -- DID HIS Ph.D. AS -- 47 00:02:11,760 --> 00:02:15,200 AT THE FAMOUS UNIVERSITY THERE, 48 00:02:15,200 --> 00:02:17,440 SEOUL NATIONAL UNIVERSITY. 49 00:02:17,440 --> 00:02:19,720 THEN HE SPENT FIVE YEARS IN MY 50 00:02:19,720 --> 00:02:25,720 LAB VERY PRODUCTIVE AND TERRIFIC 51 00:02:25,720 --> 00:02:27,080 POST-DOC IN THOSE YEARS DOING 52 00:02:27,080 --> 00:02:30,040 MANY PROJECTS THAT INCLUDED 53 00:02:30,040 --> 00:02:32,280 CHROMATIN STRUCTURAL ASPECTS 54 00:02:32,280 --> 00:02:35,400 THAT INCLUDED EPIGENETIC 55 00:02:35,400 --> 00:02:42,320 STUDIES, AND HE HAS BEEN VERY 56 00:02:42,320 --> 00:02:44,360 EXCELLENT COLLEAGUE IN THE LAB. 57 00:02:44,360 --> 00:02:47,680 I'M PLEASED HE GOT A GREAT JOB 58 00:02:47,680 --> 00:02:51,360 WHERE HE IS NOW ASSISTANT 59 00:02:51,360 --> 00:02:52,000 PROFESSOR AT MERCER UNIVERSITY 60 00:02:52,000 --> 00:02:52,840 SCHOOL OF MEDICINE AND HE 61 00:02:52,840 --> 00:02:56,120 STARTED THIS YEAR. 62 00:02:56,120 --> 00:03:01,080 HE WILL TALK ABOUT A PROJECT HE 63 00:03:01,080 --> 00:03:04,600 ORGANIZED ONE IN MY LAB ABOUT 64 00:03:04,600 --> 00:03:05,800 MITOCHONDRIAL CORRELATION. 65 00:03:05,800 --> 00:03:08,840 SO JONG, PLEASE TALK. 66 00:03:08,840 --> 00:03:15,080 THANK YOU, DOCTOR BOHR FOR YOUR 67 00:03:15,080 --> 00:03:15,680 INTRODUCTION. 68 00:03:15,680 --> 00:03:17,440 HELLO EVERYONE. 69 00:03:17,440 --> 00:03:20,040 I'M JONG HYUK LEE, ASSISTANT 70 00:03:20,040 --> 00:03:20,840 PROFESSOR AT MERCER UNIVERSITY 71 00:03:20,840 --> 00:03:22,040 SCHOOL OF MEDICINE. 72 00:03:22,040 --> 00:03:24,160 HERE IN GEORGIA, SAVANNAH. 73 00:03:24,160 --> 00:03:25,720 TODAY I'M GOING TO PRESENT ONE 74 00:03:25,720 --> 00:03:29,200 OF MY RESEARCH PROJECTS ENTITLED 75 00:03:29,200 --> 00:03:30,560 MITOCHONDRIAL PARP1 REGULATES 76 00:03:30,560 --> 00:03:31,880 NAD DEPENDENT POLYADP 77 00:03:31,880 --> 00:03:32,320 RIBOSYLATION ON MITOCHONDRIAL 78 00:03:32,320 --> 00:03:35,760 NUCLEOID. 79 00:03:35,760 --> 00:03:37,080 THIS WORK WAS MOSTLY DONE WHILE 80 00:03:37,080 --> 00:03:39,080 I WAS DOING MY POST-DOCTORAL 81 00:03:39,080 --> 00:03:40,280 RESEARCH AT NATIONAL INSTITUTE 82 00:03:40,280 --> 00:03:46,360 ON AGING IN DR. BORE'S LAB. 83 00:03:46,360 --> 00:03:49,240 IT IS KNOWN DNA DAMAGE CAUSES 84 00:03:49,240 --> 00:03:50,800 CASCADE OF SIGNAL TRANSDUCTION 85 00:03:50,800 --> 00:03:52,760 WHICH LEADS TO 86 00:03:52,760 --> 00:03:54,360 NEURODEGENERATION, CANCER AND 87 00:03:54,360 --> 00:03:55,080 AGING. 88 00:03:55,080 --> 00:03:57,680 AMONG THESE SIGNAL TRANSDUCERS, 89 00:03:57,680 --> 00:04:01,120 DNA DAMAGE INDUCED PARP 90 00:04:01,120 --> 00:04:03,280 ACTIVATION RESULTING 91 00:04:03,280 --> 00:04:04,760 INTRACELLULAR NAD DEPRIVATION IS 92 00:04:04,760 --> 00:04:08,480 RESPONSIBLE FOR MITOCHONDRIAL 93 00:04:08,480 --> 00:04:11,120 DYSFUNCTION. 94 00:04:11,120 --> 00:04:15,120 PARP1 IS VERY IMPORTANT IN 95 00:04:15,120 --> 00:04:16,000 MAINTAINING GENE INTEGRITY. 96 00:04:16,000 --> 00:04:19,200 IT PLAYS A CRITICAL ROLE IN MANY 97 00:04:19,200 --> 00:04:20,760 NUCLEAR DNA REPAIR PROCESSES. 98 00:04:20,760 --> 00:04:24,560 IN ADDITION TO THIS THERE ARE 99 00:04:24,560 --> 00:04:27,120 NUMBER OF REPORTS PARP AFFECTS 100 00:04:27,120 --> 00:04:29,120 MITOCHONDRIAL HOMEOSTASIS. 101 00:04:29,120 --> 00:04:32,080 PARP1 INHIBITION SEEMS LIKE 102 00:04:32,080 --> 00:04:34,200 INCREASING MITOCHONDRIAL 103 00:04:34,200 --> 00:04:36,640 METABOLISM, REDUCING REACTIVE 104 00:04:36,640 --> 00:04:39,840 OXYGEN SPECIES PRODUCTION. 105 00:04:39,840 --> 00:04:44,080 AND ENHANCING MITOCHONDRIAL 106 00:04:44,080 --> 00:04:49,080 BIOGENESIS AND REPAIR ACTIVITY. 107 00:04:49,080 --> 00:04:55,920 AFTER INTRAMITOCHONDRIAL 108 00:04:55,920 --> 00:04:58,040 PARYLATION WAS FOUND IN 1975 109 00:04:58,040 --> 00:04:59,640 IT'S BEEN OBSERVE AND STUDY. 110 00:04:59,640 --> 00:05:02,520 HOW FAR MITOCHONDRIAL PARP 111 00:05:02,520 --> 00:05:04,720 INEXISTENCE WAS A MATTER OF 112 00:05:04,720 --> 00:05:07,240 DEBATE IN THE FIELD UNTIL 113 00:05:07,240 --> 00:05:07,840 RECENTLY. 114 00:05:07,840 --> 00:05:09,600 THIS IS TAKEN FROM ONE OF THE 115 00:05:09,600 --> 00:05:12,240 REVIEW PAPERS AND YOU CAN TAKE A 116 00:05:12,240 --> 00:05:15,280 LOOK WHAT PEOPLE THINK ABOUT 117 00:05:15,280 --> 00:05:20,680 MITOCHONDRIAL PARP1. 118 00:05:20,680 --> 00:05:21,280 AS FAR AS ENTERER MITOCHONDRIAL 119 00:05:21,280 --> 00:05:23,080 THE FIELD WAS LARGELY NEGLECTED 120 00:05:23,080 --> 00:05:26,680 AND NOT STUDIED SYSTEMATICALLY. 121 00:05:26,680 --> 00:05:28,080 MOREOVER WHATEVER LITTLE WORK 122 00:05:28,080 --> 00:05:32,160 FORMED IN MITOCHONDRIAL PARP IT 123 00:05:32,160 --> 00:05:33,640 DIVERGES, SOME INVESTIGATORS 124 00:05:33,640 --> 00:05:36,880 FIND IT WHILE OTHERS DON'T. 125 00:05:36,880 --> 00:05:39,200 SO WE CONCLUDED THAT THE REASON 126 00:05:39,200 --> 00:05:42,720 WHY PEOPLE ARE HAVING DEBATES 127 00:05:42,720 --> 00:05:45,960 ABOUT MITOCHONDRIAL PARP 128 00:05:45,960 --> 00:05:47,280 EXISTING IS SIMPLY BECAUSE IT IS 129 00:05:47,280 --> 00:05:49,960 NOT STUDIED SYSTEMATICALLY. 130 00:05:49,960 --> 00:05:52,680 SO WE TRY TO SOLVE THIS BY 131 00:05:52,680 --> 00:05:57,560 STUDYING IT SYSTEMATICALLY. 132 00:05:57,560 --> 00:06:01,440 FIRST WE PERFORM MITOCHONDRIAL 133 00:06:01,440 --> 00:06:04,520 FRACTIONATION, MITOCHONDRIAL 134 00:06:04,520 --> 00:06:05,600 PURIFICATION FROM WHY WOULD TYPE 135 00:06:05,600 --> 00:06:09,040 AND NOT PARP KNOCK OUT AND 136 00:06:09,040 --> 00:06:09,960 INVESTIGATED PURITY BY WESTERN 137 00:06:09,960 --> 00:06:12,400 BLOT. 138 00:06:12,400 --> 00:06:17,480 OUR MITOCHONDRIAL SHOWS ABSENCE 139 00:06:17,480 --> 00:06:18,720 INDICATING FREE OF SITE 140 00:06:18,720 --> 00:06:19,440 CONTAMINATION. 141 00:06:19,440 --> 00:06:23,760 OUR SAMPLE WAS ALSO FREE OF 142 00:06:23,760 --> 00:06:26,320 LAMIN A AND HISTONE H 3 SHOWING 143 00:06:26,320 --> 00:06:30,400 NUCLEAR CONTAMINATION AS WELL. 144 00:06:30,400 --> 00:06:33,400 IN THIS CONDITION WE COULD FIND 145 00:06:33,400 --> 00:06:36,720 PARP EXISTENCE IN PURIFIED 146 00:06:36,720 --> 00:06:38,280 MITOCHONDRIAL FROM WILD TYPE 147 00:06:38,280 --> 00:06:39,440 CELLS BUT NOT PARP1 KNOCK OUT 148 00:06:39,440 --> 00:06:41,600 CELLS. 149 00:06:41,600 --> 00:06:44,400 WE THEN WANTED TO MAKE SURE THIS 150 00:06:44,400 --> 00:06:46,080 FINDING IS NOT COMING FROM THE 151 00:06:46,080 --> 00:06:53,520 OUTER MITOCHONDRIAL MEMBRANE 152 00:06:53,520 --> 00:06:55,880 WITH PARP1 CONTAMINATION. 153 00:06:55,880 --> 00:06:58,240 WE SPLIT THE MITOCHONDRIAL OUTER 154 00:06:58,240 --> 00:07:00,560 MEMBRANE WITH OPTIMIZED PROTEIN 155 00:07:00,560 --> 00:07:01,080 K TREATMENT. 156 00:07:01,080 --> 00:07:05,880 AS YOU CAN SEE HERE, PROTEIN K 157 00:07:05,880 --> 00:07:09,520 TREATMENT REMOVED TOM 20, OUTER 158 00:07:09,520 --> 00:07:10,560 MITOCHONDRIAL MEMBRANE MARKER 159 00:07:10,560 --> 00:07:14,960 AND MAINTAINED HSP 60 WHICH IS A 160 00:07:14,960 --> 00:07:17,160 MITOCHONDRIAL MATRIX MARKER, IT 161 00:07:17,160 --> 00:07:18,880 CAN STILL FIND PARP1 IN THIS 162 00:07:18,880 --> 00:07:20,840 CONDITION SUCH AS IN THE PARP1 163 00:07:20,840 --> 00:07:22,560 LOCALIZING IN THE MITOCHONDRIAL 164 00:07:22,560 --> 00:07:24,960 MATRIX. 165 00:07:24,960 --> 00:07:27,040 WE ONCE AGAIN PERFORMED 166 00:07:27,040 --> 00:07:29,560 MITOCHONDRIAL FRACTIONATION TO 167 00:07:29,560 --> 00:07:33,240 INVESTIGATE PARP1 LOCALIZATION 168 00:07:33,240 --> 00:07:35,600 IN INNER MITOCHONDRIAL MEMBRANE 169 00:07:35,600 --> 00:07:37,800 AND OUTER MITOCHONDRIAL MEMBRANE 170 00:07:37,800 --> 00:07:40,840 AND MITOCHONDRIAL MATRIX. 171 00:07:40,840 --> 00:07:43,160 WE FOUND SOME AMOUNT OF PARP1 IN 172 00:07:43,160 --> 00:07:45,720 THE INNER MEMBRANE OF THE 173 00:07:45,720 --> 00:07:47,560 FRACTION BUT MOST AT THE SIGNAL 174 00:07:47,560 --> 00:07:52,800 WERE FROM MITOCHONDRIAL MATRIX. 175 00:07:52,800 --> 00:07:54,800 SINCE WE FOUND PARP1 176 00:07:54,800 --> 00:07:56,640 LOCALIZATION IN THE 177 00:07:56,640 --> 00:07:58,520 MITOCHONDRIAL MATRIX WE WANTED 178 00:07:58,520 --> 00:08:03,080 TO TEST PARP1 CONFINED TO 179 00:08:03,080 --> 00:08:04,840 MITOCHONDRIAL DNA NUCLEAR DNA. 180 00:08:04,840 --> 00:08:06,920 IN ORDER TO DO THIS WE TESTED 181 00:08:06,920 --> 00:08:08,720 OUR MITOCHONDRIAL PURITY BY REAL 182 00:08:08,720 --> 00:08:13,160 TIME PCR WITH MITOCHONDRIA AND 183 00:08:13,160 --> 00:08:16,200 NUCLEAR DNA SPECIFIC PRIMERS. 184 00:08:16,200 --> 00:08:18,200 OUR MITOCHONDRIAL APPROXIMATE 185 00:08:18,200 --> 00:08:20,320 RATION WERE ALMOST FREE OF 186 00:08:20,320 --> 00:08:23,080 NUCLEAR DNA CONTAMINATION. 187 00:08:23,080 --> 00:08:26,400 FINALLY WE PERFORM PARP1 FROM 188 00:08:26,400 --> 00:08:27,800 IMMUNOPRECIPITATION USING 189 00:08:27,800 --> 00:08:29,680 PURIFIED MITOCHONDRIA AND 190 00:08:29,680 --> 00:08:33,040 MITOCHONDRIAL SPECIFIC MARKERS. 191 00:08:33,040 --> 00:08:35,160 WE WERE ABLE TO FIND PARP1 192 00:08:35,160 --> 00:08:36,280 BINDING TO THE MITOCHONDRIAL 193 00:08:36,280 --> 00:08:39,680 DNA. 194 00:08:39,680 --> 00:08:44,400 NEXT WE TESTED IN WHICH 195 00:08:44,400 --> 00:08:47,480 CONDITIONS MITOCHONDRIAL 196 00:08:47,480 --> 00:08:50,280 PARLATION IS ACTIVATED. 197 00:08:50,280 --> 00:08:52,720 NAD IS PURIFIED GLYCOSYLATION 198 00:08:52,720 --> 00:08:55,520 AND TREATMENT OF NAD TO PURIFIED 199 00:08:55,520 --> 00:08:58,200 MITOCHONDRIA GREATLY INDUCED 200 00:08:58,200 --> 00:09:02,840 MITOCHONDRIAL PARLATION, THERE 201 00:09:02,840 --> 00:09:04,480 ARE REPORTS SAYING STRESS IS 202 00:09:04,480 --> 00:09:07,840 RESPONSIBLE FOR MITOCHONDRIAL 203 00:09:07,840 --> 00:09:10,040 PARLATION. 204 00:09:10,040 --> 00:09:12,480 HYDROGEN PEROXIDE TREATMENT DID 205 00:09:12,480 --> 00:09:15,920 INDUCE MITOCHONDRIA CORRELATION. 206 00:09:15,920 --> 00:09:21,680 PARP1 INHIBITED WITH TREATMENT 207 00:09:21,680 --> 00:09:26,240 NAD INDUCED PARYLATION WAS NOT 208 00:09:26,240 --> 00:09:30,080 INDUCED INDICATING PARP1 IS 209 00:09:30,080 --> 00:09:31,760 RESPONSIBLE IN THESE REGIONS. 210 00:09:31,760 --> 00:09:33,720 WHEN YOU PERFORM THE SAME 211 00:09:33,720 --> 00:09:35,760 EXPERIMENTS WITH PURIFIED 212 00:09:35,760 --> 00:09:37,280 MITOCHONDRIA FROM KNOCK OUT 213 00:09:37,280 --> 00:09:40,520 CELLS NAD COULDN'T INDUCE 214 00:09:40,520 --> 00:09:41,520 MITOCHONDRIAL CORRELATION. 215 00:09:41,520 --> 00:09:43,480 IT SUPPORTS OUR HYPOTHESIS THAT 216 00:09:43,480 --> 00:09:46,200 PARP1 IS RESPONSIBLE FOR NAD 217 00:09:46,200 --> 00:09:47,200 INDUCED MITOCHONDRIAL 218 00:09:47,200 --> 00:09:49,880 CORRELATION. 219 00:09:49,880 --> 00:09:52,120 -- PARYLATION. 220 00:09:52,120 --> 00:09:55,720 NEXT WE TESTED IF OTHER NAD 221 00:09:55,720 --> 00:09:57,160 PRECURSORS HAVE IMPACT ON 222 00:09:57,160 --> 00:09:59,200 MITOCHONDRIAL CORRELATION. 223 00:09:59,200 --> 00:10:04,960 WE TESTED NICOTINAMIDE, 224 00:10:04,960 --> 00:10:06,920 NICOTINAMIDE MONONUCLEAR SIDE 225 00:10:06,920 --> 00:10:09,560 AND RIBOSIDE. 226 00:10:09,560 --> 00:10:12,640 NAD EXCLUSIVELY INDUCES 227 00:10:12,640 --> 00:10:14,000 PARYLATION OF PURIFIED 228 00:10:14,000 --> 00:10:17,120 MITOCHONDRIA. 229 00:10:17,120 --> 00:10:19,080 WITH THE HELP FROM OUR 230 00:10:19,080 --> 00:10:21,680 COLLABORATOR, DR. ANTHONY AT 231 00:10:21,680 --> 00:10:23,360 JOUNCES HOPKINS WE EVALUATED 232 00:10:23,360 --> 00:10:27,600 MITOCHONDRIAL PARYLATION BY 233 00:10:27,600 --> 00:10:28,560 ELTA. 234 00:10:28,560 --> 00:10:34,680 THAT IS ENZYME@ICS LABELING BY 235 00:10:34,680 --> 00:10:38,040 RIBOSE IT LABELS POLYMERS WITH 236 00:10:38,040 --> 00:10:40,920 SIFI AT THE TERMINAL PARP BRANCH 237 00:10:40,920 --> 00:10:42,720 AND CONSIDERED SPECIFIC AND 238 00:10:42,720 --> 00:10:44,880 SENSITIVE FOR DETECTING FOLIATED 239 00:10:44,880 --> 00:10:46,280 GLYCOSYLATION. 240 00:10:46,280 --> 00:10:49,880 WITH THIS ELT LABELING WE COULD 241 00:10:49,880 --> 00:10:53,360 FIND MITOCHONDRIAL PARYLATION. 242 00:10:53,360 --> 00:10:56,000 CONSISTENT WITH EARLIER FINDINGS 243 00:10:56,000 --> 00:10:57,480 OUR TREATMENT SUPPRESSES 244 00:10:57,480 --> 00:11:00,240 MITOCHONDRIAL PARYLATION AND 245 00:11:00,240 --> 00:11:01,440 SUPPORTS PARP1 INVOLVEMENT IN 246 00:11:01,440 --> 00:11:06,280 THIS REACTION. 247 00:11:06,280 --> 00:11:09,360 IN ORDER TO VERIFY PAR 1 248 00:11:09,360 --> 00:11:12,000 MEDIATED MITOCHONDRIAL 249 00:11:12,000 --> 00:11:13,160 PARYLATION WE ADOPTED ANOTHER 250 00:11:13,160 --> 00:11:16,480 TECHNIQUE CALLED ADPR PROTEIN 251 00:11:16,480 --> 00:11:19,480 AFFINITY PURIFICATION. 252 00:11:19,480 --> 00:11:22,560 IT IS VERY SIMILAR TECHNIQUE TO 253 00:11:22,560 --> 00:11:23,880 TRADITIONAL CHROMATIN 254 00:11:23,880 --> 00:11:24,840 IMMUNOPRECIPITATION. 255 00:11:24,840 --> 00:11:29,120 HOWEVER, IT USES FOLIATED RIBOSE 256 00:11:29,120 --> 00:11:31,320 SPECIFIC BINDING PROTEIN CALLED 257 00:11:31,320 --> 00:11:34,960 WWE INSTEAD OF ANTIBODY IN 258 00:11:34,960 --> 00:11:37,680 REACTIONS SO WITH THESE ADPR 259 00:11:37,680 --> 00:11:39,640 CHAT WE CAN PUT OUT CORRELATED 260 00:11:39,640 --> 00:11:41,840 CHROMATIN AND ANALYZE BY DEEP 261 00:11:41,840 --> 00:11:45,000 SEQUENCE OR REAL TIME PCR. 262 00:11:45,000 --> 00:11:47,240 CONSISTENT WITH THE WESTERN BLOT 263 00:11:47,240 --> 00:11:51,440 RESULT WE COULD FIND NAD INDUCED 264 00:11:51,440 --> 00:11:52,640 PAYYLATION AND MEDIATED 265 00:11:52,640 --> 00:11:56,040 SUPPRESSION OF MITOCHONDRIAL 266 00:11:56,040 --> 00:11:57,280 PARYLATION IN WILD TYPE CELLS 267 00:11:57,280 --> 00:11:59,920 BUT NOT PARP1 KNOCK OUT CELLS. 268 00:11:59,920 --> 00:12:02,480 THIS AGAIN SUPPORTING PARP1 269 00:12:02,480 --> 00:12:04,720 RESPONSIBLE FOR NAD INDUCED 270 00:12:04,720 --> 00:12:09,360 MITOCHONDRIAL PAYYLATION IN 271 00:12:09,360 --> 00:12:11,840 PURIFIED MITOCHONDRIA. 272 00:12:11,840 --> 00:12:14,240 WE ALSO SUBJECTED PURIFIED DNA 273 00:12:14,240 --> 00:12:19,120 FROM APR TO DNA SEQUENCING. 274 00:12:19,120 --> 00:12:23,520 AND GENERATED PAYYLATION MAP ON 275 00:12:23,520 --> 00:12:24,360 MITOCHONDRIAL GENOME. 276 00:12:24,360 --> 00:12:26,280 PARP DISTRIBUTION SHOWED 277 00:12:26,280 --> 00:12:28,400 DISPERSE ENRICHMENT OVER 278 00:12:28,400 --> 00:12:29,400 MITOCHONDRIAL GENOME. 279 00:12:29,400 --> 00:12:35,920 IT IS A BIT DENSE IN REGIONS AND 280 00:12:35,920 --> 00:12:37,280 SCARCE IN (INAUDIBLE) REGIONS. 281 00:12:37,280 --> 00:12:39,680 THE OVERALL PARP DISTRIBUTION ON 282 00:12:39,680 --> 00:12:41,480 MITOCHONDRIAL GENE LOCUST WAS 283 00:12:41,480 --> 00:12:43,120 CONSISTENT WITH OUR EARLIER 284 00:12:43,120 --> 00:12:47,760 FINDINGS THAT MITOCHONDRIAL 285 00:12:47,760 --> 00:12:49,480 PAYYLATION WAS INDUCED BY NAD 286 00:12:49,480 --> 00:12:51,440 TREATMENT AND SUPPRESSED BY 287 00:12:51,440 --> 00:12:51,840 TREATMENT. 288 00:12:51,840 --> 00:12:53,960 IT WAS NOT HAPPENING IN PURIFIED 289 00:12:53,960 --> 00:12:55,760 MITOCHONDRIA FROM PARP1 KNOCK 290 00:12:55,760 --> 00:13:00,840 OUT CELLS. 291 00:13:00,840 --> 00:13:03,600 IT IS KNOWN HISTONE DENSITY IS 292 00:13:03,600 --> 00:13:10,480 CLOSELY RELATED TO THE CHROMATIN 293 00:13:10,480 --> 00:13:16,920 PARYLATION IN NUCLEUS SO WE 294 00:13:16,920 --> 00:13:19,520 HYPOTHESIZED IT WAS IN NUCLEOID. 295 00:13:19,520 --> 00:13:21,760 IT DOES HAVE -- WHICH STABILIZES 296 00:13:21,760 --> 00:13:23,800 MITOCHONDRIAL DNA BY PACKAGING 297 00:13:23,800 --> 00:13:27,000 INTO MITOCHONDRIAL NUCLEOIDS. 298 00:13:27,000 --> 00:13:30,880 IN ORDER TO TEST KEY FACTORS 299 00:13:30,880 --> 00:13:34,280 INVOLVED IN MITOCHONDRIAL 300 00:13:34,280 --> 00:13:38,160 NUCLEOID PARYLATION WE PERFORMED 301 00:13:38,160 --> 00:13:40,840 SEQUENCE ANALYSIS ON CHIP AND 302 00:13:40,840 --> 00:13:43,840 GAP PURIFIED DNA. 303 00:13:43,840 --> 00:13:47,320 AS A RESULT PARP DISTRIBUTION ON 304 00:13:47,320 --> 00:13:49,040 MITOCHONDRIA GENOME SHOW 305 00:13:49,040 --> 00:13:52,040 STRIKINGLY SIMILAR PATTERN. 306 00:13:52,040 --> 00:13:55,200 AND WHEN CHIP AND PARP 307 00:13:55,200 --> 00:13:57,760 SEQUENCING SIGNALING INTENSITY 308 00:13:57,760 --> 00:14:00,440 ON 37 MITOCHONDRIAL GENES, IT 309 00:14:00,440 --> 00:14:02,280 SHOWED VERY NICE POSITIVE 310 00:14:02,280 --> 00:14:07,200 CORRELATION. 311 00:14:07,200 --> 00:14:10,400 WE THEN PERFORMED KEY AND CHIP 312 00:14:10,400 --> 00:14:12,600 WITH PURIFIED MITOCHONDRIA TO 313 00:14:12,600 --> 00:14:14,080 EVALUATE FINDING. 314 00:14:14,080 --> 00:14:17,840 WE FOUND INCREASED TFAM 315 00:14:17,840 --> 00:14:20,360 OCCUPANCY ON THE REGION OF 316 00:14:20,360 --> 00:14:23,080 MITOCHONDRIAL NUCLEOID WITH NAD 317 00:14:23,080 --> 00:14:26,600 TREATMENT, VERY SIMILAR FOR THE 318 00:14:26,600 --> 00:14:27,640 PARYLATION PATTERN. 319 00:14:27,640 --> 00:14:29,840 IT IS ONCE OF THE MITOCHONDRIA 320 00:14:29,840 --> 00:14:32,320 TRANSCRIPTION FACTORS KNOWN TO 321 00:14:32,320 --> 00:14:34,040 INDUCE TRANSCRIPTION WHEN BOUND 322 00:14:34,040 --> 00:14:37,360 TO MITOCHONDRIAL REGION. 323 00:14:37,360 --> 00:14:41,720 WE HYPOTHESIZE THAT NAD INDUCED 324 00:14:41,720 --> 00:14:43,840 EQUIPMENT TO THE REGION MAY 325 00:14:43,840 --> 00:14:46,680 AFFECT MITOCHONDRIAL DNA 326 00:14:46,680 --> 00:14:47,080 TRANSCRIPTION. 327 00:14:47,080 --> 00:14:48,880 WHEN CHECKING MITOCHONDRIAL GENE 328 00:14:48,880 --> 00:14:51,080 EXPRESSION WE FOUND THAT 329 00:14:51,080 --> 00:14:52,360 MITOCHONDRIAL TRANSCRIPTION IS 330 00:14:52,360 --> 00:14:54,760 SIGNIFICANTLY INDUCED WITH NAD 331 00:14:54,760 --> 00:14:58,280 TREATMENT IN WILD TYPE CELLS, 332 00:14:58,280 --> 00:15:01,720 BUT NOT MUCH IN PARP. 333 00:15:01,720 --> 00:15:05,040 THIS SUGGEST NAD INDUCED TFAM 334 00:15:05,040 --> 00:15:07,240 TREATMENT TO DELOOP REGION 335 00:15:07,240 --> 00:15:08,600 REGULATES MITOCHONDRIAL 336 00:15:08,600 --> 00:15:10,600 TRANSCRIPTION. 337 00:15:10,600 --> 00:15:12,040 THIS IS THE SUMMARY OF OUR 338 00:15:12,040 --> 00:15:15,280 FINDINGS. 339 00:15:15,280 --> 00:15:18,160 WE CONFIRM MITOCHONDRIAL PARP1 340 00:15:18,160 --> 00:15:22,280 AND PARYLATION BY MULTIPLE 341 00:15:22,280 --> 00:15:26,520 SETTINGS, SUCH AS WESTERN BLOT, 342 00:15:26,520 --> 00:15:27,880 IMMUNOPRECIPITATION, ADP 343 00:15:27,880 --> 00:15:29,200 AFFINITY CHROMATIN PURIFICATION 344 00:15:29,200 --> 00:15:32,080 AND ELPA OF WE FOUND NAD 345 00:15:32,080 --> 00:15:35,360 TREATMENT STIMULATED PARP1 346 00:15:35,360 --> 00:15:42,200 MEDIATED MITOCHONDRIAL NUCLEOID 347 00:15:42,200 --> 00:15:44,240 PARYLATION REGULATING TO THE D 348 00:15:44,240 --> 00:15:46,680 LOOP TO REGULATE MITOCHONDRIAL 349 00:15:46,680 --> 00:15:47,400 TRANSCRIPTION. 350 00:15:47,400 --> 00:15:50,280 I WOULD LIKE TO THANK MY 351 00:15:50,280 --> 00:15:53,040 POST-DOC MENTOR DR. (INAUDIBLE) 352 00:15:53,040 --> 00:15:54,880 FOR THEIR EVER LASTING SUPPORT. 353 00:15:54,880 --> 00:15:58,680 I WOULD LIKE TO THANK MY 354 00:15:58,680 --> 00:15:59,840 (INDISCERNIBLE) WHEN I WAS DOING 355 00:15:59,840 --> 00:16:02,000 THIS WORK AT NIA. 356 00:16:02,000 --> 00:16:04,640 I WOULD ALSO LIKE TO THANK MY 357 00:16:04,640 --> 00:16:09,400 COLLABORATOR, DR. ANTHONY LEON 358 00:16:09,400 --> 00:16:11,040 AND (INDISCERNIBLE) FOR THEIR 359 00:16:11,040 --> 00:16:13,400 CONTRIBUTIONS AT JOHNS HOPKINS 360 00:16:13,400 --> 00:16:14,080 ELT EXPENSE. 361 00:16:14,080 --> 00:16:17,040 THANK YOU FOR YOUR ATTENTION. 362 00:16:17,040 --> 00:16:19,120 >> THANK YOU FOR THAT WONDERFUL 363 00:16:19,120 --> 00:16:19,640 TALK. 364 00:16:19,640 --> 00:16:22,680 WE WILL HAVE THE QUESTIONS AT 365 00:16:22,680 --> 00:16:26,640 THE END. 366 00:16:26,640 --> 00:16:30,000 RIGHT NOW GOING TO UNIVERSITY OF 367 00:16:30,000 --> 00:16:31,840 PITTSBURGH, MATT SCHAICH WILL BE 368 00:16:31,840 --> 00:16:32,840 INTRODUCED. 369 00:16:32,840 --> 00:16:35,960 MY PLEASURE TO INTRODUCE MATT 370 00:16:35,960 --> 00:16:36,480 SCHAICH. 371 00:16:36,480 --> 00:16:37,920 HE RECEIVED BACHELORS 372 00:16:37,920 --> 00:16:39,120 BIOCHEMISTRY AND MUSIC 373 00:16:39,120 --> 00:16:43,760 UNIVERSITY OF NEBRASKA LINCOLN 374 00:16:43,760 --> 00:16:45,600 IN 2014 AND FROM THERE WENT TO 375 00:16:45,600 --> 00:16:47,040 (INAUDIBLE) LAB AT KANSAS 376 00:16:47,040 --> 00:16:48,640 UNIVERSITY MEDICAL CENTER. 377 00:16:48,640 --> 00:16:51,920 THERE HE SOLVED SEVERAL 378 00:16:51,920 --> 00:16:53,240 STRUCTURE POLYMERASE HOMOLOGUE 379 00:16:53,240 --> 00:16:54,640 AND IDENTIFIED INTERESTING 380 00:16:54,640 --> 00:16:57,600 RESIDUE THAT ALLOWS POLYMERASE 381 00:16:57,600 --> 00:17:00,160 TO SELECT AGAINST INSERTION OF 382 00:17:00,160 --> 00:17:00,880 RIBONUCLEOTIDES. 383 00:17:00,880 --> 00:17:04,120 HE MOVED TO THE CANCER CENTER 384 00:17:04,120 --> 00:17:05,680 2020, JOINED BEN'S LAB AFTER HE 385 00:17:05,680 --> 00:17:07,320 SUCCESSFULLY COMPETED FOR ONE OF 386 00:17:07,320 --> 00:17:09,560 THESE HIGHLY COMPETITIVE 387 00:17:09,560 --> 00:17:10,800 FELLOWSHIPS FOR INNOVATIVE 388 00:17:10,800 --> 00:17:11,360 CANCER RESEARCH. 389 00:17:11,360 --> 00:17:14,320 HE'S BEEN HIGHLY PRODUCTIVE AND 390 00:17:14,320 --> 00:17:18,440 HE HAS MASTERED THIS OPTICAL 391 00:17:18,440 --> 00:17:19,040 TWEEZER INSTRUMENT. 392 00:17:19,040 --> 00:17:20,400 LONG TIME WITH THIS INSTRUMENT 393 00:17:20,400 --> 00:17:22,240 DOING SINGLE MOLECULE STUDIES SO 394 00:17:22,240 --> 00:17:24,600 I THINK YOU ARE IN FOR A REAL 395 00:17:24,600 --> 00:17:25,840 TREAT, VERY EXCITING DATA TO 396 00:17:25,840 --> 00:17:26,160 SHARE. 397 00:17:26,160 --> 00:17:27,800 GO AHEAD MATT. 398 00:17:27,800 --> 00:17:28,440 >> EXCELLENT. 399 00:17:28,440 --> 00:17:30,560 THANK YOU SO MUCH FOR THAT 400 00:17:30,560 --> 00:17:31,080 INTRODUCTION. 401 00:17:31,080 --> 00:17:33,160 I'M REALLY EXCITEDDED TO TALK 402 00:17:33,160 --> 00:17:35,440 ABOUT THIS NEW MOLECULE THAT WE 403 00:17:35,440 --> 00:17:38,920 HAVE DEVELOPED, WE USE IT AS 404 00:17:38,920 --> 00:17:41,760 OPTICALS, I'M POST-DOCTORAL 405 00:17:41,760 --> 00:17:43,360 FELLOW IN DR. (INDISCERNIBLE) 406 00:17:43,360 --> 00:17:43,800 LAB. 407 00:17:43,800 --> 00:17:45,160 I HAVE SHOWN HERE THE EXCITING 408 00:17:45,160 --> 00:17:46,720 DATA WE ARE GETTING AND I WILL 409 00:17:46,720 --> 00:17:48,600 GO OVER WHAT IT IS RED AND PINK 410 00:17:48,600 --> 00:17:50,520 AND BLUE LINES MEAN AND HOW WE 411 00:17:50,520 --> 00:17:53,680 INTERPRET HOW DNA REPAIR WORKS 412 00:17:53,680 --> 00:17:56,840 BASED ON THESE INTEGERS. 413 00:17:56,840 --> 00:17:58,560 FIRST I WANT TO TALK ABOUT ONE 414 00:17:58,560 --> 00:17:59,840 OF THE MAIN POINTS OF FOCUS OF 415 00:17:59,840 --> 00:18:06,080 THE LAB WHICH IS THAT UBDDB IS 416 00:18:06,080 --> 00:18:07,280 UV DAMAGE DNA BINDING PROTEIN 417 00:18:07,280 --> 00:18:09,920 AND A LOT OF WORK OUT OF THE VAN 418 00:18:09,920 --> 00:18:11,560 OUTEN LAB HAS SHOWN THAT REALLY 419 00:18:11,560 --> 00:18:13,520 MAYBE WE SHOULD BE CALLING IT 420 00:18:13,520 --> 00:18:15,400 DDB BECAUSE NOT ONLY DOES IT 421 00:18:15,400 --> 00:18:19,240 BIND UV DAMAGE IT ALSO BINDS 422 00:18:19,240 --> 00:18:22,440 OXIDATIVE DAMAGE, ABBEY SICK 423 00:18:22,440 --> 00:18:24,400 TYPES VAST REPERTOIRE OF 424 00:18:24,400 --> 00:18:26,680 SUBSTRATES. 425 00:18:26,680 --> 00:18:29,200 SO UVDDB BINDS DAMAGE DIRECTLY 426 00:18:29,200 --> 00:18:31,560 BUT ALSO HELPS DECOMPACT 427 00:18:31,560 --> 00:18:32,480 CHROMATIN IN ORDER TO MAKE 428 00:18:32,480 --> 00:18:36,400 DAMAGE MORE ACCESSIBLE TO DNA 429 00:18:36,400 --> 00:18:38,680 GLYCOSYLATION, BER PATHWAY. 430 00:18:38,680 --> 00:18:41,520 BUT ALSO HELPS RECRUIT XPC TO 431 00:18:41,520 --> 00:18:45,240 SITES OF DAMAGE AND 432 00:18:45,240 --> 00:18:46,280 REALISTICALLY AS YOU CAN SEE 433 00:18:46,280 --> 00:18:47,800 THESE PATHWAYS ARE SUPER 434 00:18:47,800 --> 00:18:49,520 COMPLICATED AND WITH ALL THESE 435 00:18:49,520 --> 00:18:51,480 NEW FACTORS COMING IN WE NEED 436 00:18:51,480 --> 00:18:53,240 SINGLE MOLECULE STUDIES TO BE 437 00:18:53,240 --> 00:18:55,600 SURE WHICH PROTEIN COMES IN AT 438 00:18:55,600 --> 00:18:57,280 WHAT POINT IN THE PATHWAY AND 439 00:18:57,280 --> 00:18:58,920 HOW LONG IT STAYS THERE. 440 00:18:58,920 --> 00:19:02,880 SO AS YOU CAN SEE WITH UVDDB 441 00:19:02,880 --> 00:19:04,320 THERE'S POINTS IT INTERACTS WITH 442 00:19:04,320 --> 00:19:05,840 THE PATHWAY, WHEN IT COMES INTO 443 00:19:05,840 --> 00:19:07,440 THE PATHWAY, WHEN IT LEAVES THE 444 00:19:07,440 --> 00:19:10,160 PATHWAY AND WE CAN HAVE A SINGLE 445 00:19:10,160 --> 00:19:11,640 POINT OF DNA DAMAGE WATCH THESE 446 00:19:11,640 --> 00:19:17,480 FACTORS COMING AND GOING WE CAN 447 00:19:17,480 --> 00:19:19,240 DEFINITIVELY SAY HOW THE 448 00:19:19,240 --> 00:19:19,880 PATHWAYS WORK. 449 00:19:19,880 --> 00:19:21,760 ONE CHALLENGE IN SINGLE MOLECULE 450 00:19:21,760 --> 00:19:23,760 STUDY IS TO HAVE ACTIVE PURIFIED 451 00:19:23,760 --> 00:19:26,120 PROTEIN THAT YOU CAN USE IN 452 00:19:26,120 --> 00:19:28,920 ORDER TO WATCH THEM AT SINGLE 453 00:19:28,920 --> 00:19:30,000 MOLECULE LEVEL AND WE WERE 454 00:19:30,000 --> 00:19:31,920 THINKING WOULDN'T IT BE GREAT IF 455 00:19:31,920 --> 00:19:33,800 INSTEAD OF GOING TO TROUBLE OF 456 00:19:33,800 --> 00:19:37,720 PURIFYING PROTEINS YOU CAN SEE 457 00:19:37,720 --> 00:19:38,920 SO MANY IN THIS PATHWAY IF WE 458 00:19:38,920 --> 00:19:41,160 CAN DIRECTLY TAKE PROTEINS OUT 459 00:19:41,160 --> 00:19:43,480 OF NUCLEAR EXTRAK AND VISUALIZE 460 00:19:43,480 --> 00:19:46,120 THEM AT SINGLE MOLECULE LEVEL. 461 00:19:46,120 --> 00:19:47,800 SO THAT IS END OF THE FROM 462 00:19:47,800 --> 00:19:50,960 APOLOGETIC TRYING TO PIONEER NEW 463 00:19:50,960 --> 00:19:52,800 METHOD WE CALL SMADNE. 464 00:19:52,800 --> 00:19:55,280 WE CALL IT SMADNE BECAUSE IT 465 00:19:55,280 --> 00:19:56,960 STANDS FOR SINGLE MOLECULE 466 00:19:56,960 --> 00:19:59,760 ANALYSIS OF DNA BINDING PROOFING 467 00:19:59,760 --> 00:20:01,080 TEEN FROM NUCLEAR EXTRACT BUT 468 00:20:01,080 --> 00:20:03,400 ALSO SHARES THE NAME WITH A BEE 469 00:20:03,400 --> 00:20:08,200 WHICH TRANSLATES THE FIRST MONK. 470 00:20:08,200 --> 00:20:10,640 LIKE THE MONK IS THIRSTY FOR 471 00:20:10,640 --> 00:20:12,320 BEER WE ARE THIRSTY FOR SINGLE 472 00:20:12,320 --> 00:20:13,720 MOLECULE DATA SO THIS ALLOWS US 473 00:20:13,720 --> 00:20:18,760 TO RAPIDLY SERVE PROTEIN 474 00:20:18,760 --> 00:20:19,800 VARIANT, WE ARE NOT GOING 475 00:20:19,800 --> 00:20:21,240 THROUGH STEPS OF PURIFICATION, 476 00:20:21,240 --> 00:20:22,920 ALL WE NEED IS INDICATION OF 477 00:20:22,920 --> 00:20:24,160 PLASMID, DIRECTLY TO SINGLE 478 00:20:24,160 --> 00:20:24,760 MOLECULE STUDY. 479 00:20:24,760 --> 00:20:27,320 WE CAN LOOK AT PTMs, PROTEIN 480 00:20:27,320 --> 00:20:29,760 BINDING KINETICS TO DNA TARGET 481 00:20:29,760 --> 00:20:31,480 AND IMPORTANTLY WE HAVE MULTIPLE 482 00:20:31,480 --> 00:20:33,160 COLORS IN THIS SET UP SO WE CAN 483 00:20:33,160 --> 00:20:35,240 LOOK AT PROTEINS COMING AND 484 00:20:35,240 --> 00:20:38,800 GOING SIMULTANEOUSLY. 485 00:20:38,800 --> 00:20:42,200 THE WORK FLOW FOR SMADNE, FIVE 486 00:20:42,200 --> 00:20:43,800 MAIN STEPS REALLY A TEAM EFFORT 487 00:20:43,800 --> 00:20:45,240 IN THE LAB SO FIRST WE HAVE A 488 00:20:45,240 --> 00:20:48,440 VECTOR WE WANT, (INAUDIBLE) 489 00:20:48,440 --> 00:20:50,200 TALENTED UNDERGRAD TECHNICIAN IN 490 00:20:50,200 --> 00:20:53,840 THE LAB, VECTORS HAND THEM OFF 491 00:20:53,840 --> 00:20:55,000 TO (INAUDIBLE) ANOTHER TALENTED 492 00:20:55,000 --> 00:20:56,960 TECHNICIAN IN THE LAB WHO 493 00:20:56,960 --> 00:20:58,640 TRANSFECT THE CELLS AND PERFORMS 494 00:20:58,640 --> 00:21:00,920 NUCLEAR EXTRACT AND PUT THEM ON 495 00:21:00,920 --> 00:21:04,040 THE MICROSCOPE WHERE WE WILL 496 00:21:04,040 --> 00:21:06,120 VISUALIZE DIFFERENT FORMS OF DNA 497 00:21:06,120 --> 00:21:07,800 DAMAGE AND WATCH THE REAL TIME 498 00:21:07,800 --> 00:21:10,480 DYNAMICS OF DNA REPAIR. 499 00:21:10,480 --> 00:21:12,400 SO WHEN I SAY MICROSCOPE WHAT DO 500 00:21:12,400 --> 00:21:13,800 I MEAN BY THAT? 501 00:21:13,800 --> 00:21:17,040 THIS IS THAT OPTICAL TWEEZER 502 00:21:17,040 --> 00:21:18,840 SEPTEMBER UP THAT WAS ALLUDED TO 503 00:21:18,840 --> 00:21:20,720 AND THIS IS HOW IT WORKS SO WE 504 00:21:20,720 --> 00:21:22,360 HAVE FIVE CHAMBER FLOW CELL 505 00:21:22,360 --> 00:21:24,480 WHICH ARE REALLY AWESOME BECAUSE 506 00:21:24,480 --> 00:21:26,680 THEY ARE MICROFLUIDIC SO WE USE 507 00:21:26,680 --> 00:21:29,640 VERY SMALL AMOUNT OF SAMPLE, WE 508 00:21:29,640 --> 00:21:33,160 ALSO HAVE A THREE COLOR CONFOCAL 509 00:21:33,160 --> 00:21:35,120 MICROSCOPE AND FIVE CHAMBERS SO 510 00:21:35,120 --> 00:21:36,560 WHEN WE ARE PULLING THE FLOW 511 00:21:36,560 --> 00:21:38,960 CELL THE LAMINAR KEEPS THEM 512 00:21:38,960 --> 00:21:41,080 SEPARATE SOD YOU CAN KEEP FIVE 513 00:21:41,080 --> 00:21:43,200 SOLUTIONS FROM MIXING, REALLY 514 00:21:43,200 --> 00:21:46,840 IMPORTANT FOR CATCHING DNA. 515 00:21:46,840 --> 00:21:50,200 SO THE FIRST STEP IN THE 516 00:21:50,200 --> 00:21:53,400 EXPERIMENT IS TO FLOW STRIP OUT 517 00:21:53,400 --> 00:21:55,960 ENCODED BEAD AND TRAP THEM INTO 518 00:21:55,960 --> 00:21:58,080 THE OPTICAL TRAP, BETWEEN ONE TO 519 00:21:58,080 --> 00:22:00,520 FOUR MICROMETERS IN SIZE SO IT 520 00:22:00,520 --> 00:22:02,160 IS TWO BEADS AND TWO TRACKS AND 521 00:22:02,160 --> 00:22:04,160 WE MOVE IT FROM CHANNEL ONE IN 522 00:22:04,160 --> 00:22:06,800 THE FLOW CELL TO 2, AND CHANNEL 523 00:22:06,800 --> 00:22:08,080 2 HAS DNA IN IT. 524 00:22:08,080 --> 00:22:12,400 WE CAN USE A BUY TIN LINKAGE TO 525 00:22:12,400 --> 00:22:14,000 STRING UP BOTH SIDES OF THE DNA 526 00:22:14,000 --> 00:22:15,960 IN THE TRACK. 527 00:22:15,960 --> 00:22:18,720 BECAUSE THIS INSTRUMENT HAS VERY 528 00:22:18,720 --> 00:22:20,200 SENSITIVE COURSE DETECTED WE CAN 529 00:22:20,200 --> 00:22:21,760 PULL THE DNA BY MOVING THE 530 00:22:21,760 --> 00:22:23,200 TRACKS IN AND OUT IN ORDER TO 531 00:22:23,200 --> 00:22:26,920 SEE AND VERIFY THAT WE HAVE ONE 532 00:22:26,920 --> 00:22:28,480 SINGLE STRAND OF DNA TETHERED 533 00:22:28,480 --> 00:22:29,440 BETWEEN THE TRACKS. 534 00:22:29,440 --> 00:22:30,720 SO SOMETIMES YOU CAN ACCIDENTLY 535 00:22:30,720 --> 00:22:32,680 CATCH TWO OR THREE BUT WE CAN 536 00:22:32,680 --> 00:22:34,040 KNOW FOR SURE WE ONLY HAVE ONE. 537 00:22:34,040 --> 00:22:36,160 ALSO WE KNOW EXACTLY WHERE IT IS 538 00:22:36,160 --> 00:22:39,600 BECAUSE IT GOES RIGHT TO CENTER. 539 00:22:39,600 --> 00:22:41,600 WE THEN WASH IT OFF AND GO 540 00:22:41,600 --> 00:22:45,120 DIRECTLY INTO NUCLEAR EXTRACT OF 541 00:22:45,120 --> 00:22:46,360 INTEREST. 542 00:22:46,360 --> 00:22:48,160 WE CAN THEN WASH THE DYNAMIC OF 543 00:22:48,160 --> 00:22:49,160 THESE EXTRACT BINDING TO DNA 544 00:22:49,160 --> 00:22:51,560 TARGET. 545 00:22:51,560 --> 00:22:54,320 SO THIS IS IN A 2D SCAN BINDING 546 00:22:54,320 --> 00:22:55,680 THAT LOOKS LIKE A CIRCLE BUT 547 00:22:55,680 --> 00:22:57,520 OFTEN WHAT WE LIKE TO DO IS 548 00:22:57,520 --> 00:22:58,720 INSTEAD OF SCANNING ALL THE 549 00:22:58,720 --> 00:23:00,200 BEADS OVER AND OVER WE WILL ONLY 550 00:23:00,200 --> 00:23:02,960 SCAN ALONG THE BLUE LINE. 551 00:23:02,960 --> 00:23:04,400 SO THEN INSTEAD OF CIRCLE IT 552 00:23:04,400 --> 00:23:06,080 LOOKS LIKE A LINE BECAUSE WHEN 553 00:23:06,080 --> 00:23:07,920 PROTEIN COMES ON LINE COMES ON 554 00:23:07,920 --> 00:23:11,080 AND THEN IF IT IS NOT MOVING, 555 00:23:11,080 --> 00:23:15,120 STAYING DOWN -- LOOKS LIKE A 556 00:23:15,120 --> 00:23:17,160 HORIZONTAL LINE. 557 00:23:17,160 --> 00:23:19,800 SO I WOULD LIKE TO BACKTRACK AND 558 00:23:19,800 --> 00:23:22,560 REVISIT UVDDB AND TALK ABOUT 559 00:23:22,560 --> 00:23:23,800 CANONICAL BEFORE WE TALK ABOUT 560 00:23:23,800 --> 00:23:24,160 RESULTS. 561 00:23:24,160 --> 00:23:28,360 THE UVDBB STANDS FOR UV DNA 562 00:23:28,360 --> 00:23:30,400 DAMAGE BINDING PROTEIN AND 563 00:23:30,400 --> 00:23:31,960 INTERACTS WITH STRONG PRODUCTS 564 00:23:31,960 --> 00:23:34,480 BUT ALSO OTHER SUBSTRATES LIKE 565 00:23:34,480 --> 00:23:36,880 BASIC -- TWO SUBUNITS HERE, I 566 00:23:36,880 --> 00:23:39,480 HAVE A COMPOSITE OF TWO 567 00:23:39,480 --> 00:23:42,520 STRUCTURE CRYSTAL AND CRYOEM 568 00:23:42,520 --> 00:23:47,160 SHOWING UVDDB BINDS TO 569 00:23:47,160 --> 00:23:50,760 NUCLEOSOME WITH DAMAGE. 570 00:23:50,760 --> 00:23:53,520 ZERO PIGMENTOSA PATIENTS HAVE 571 00:23:53,520 --> 00:23:57,280 UVDDB 2 AND KNOCK OUTS 572 00:23:57,280 --> 00:23:58,560 SPONTANEOUS TUMOR. 573 00:23:58,560 --> 00:24:01,000 IT IS IMPORTANT TO MENTION DDB 1 574 00:24:01,000 --> 00:24:04,840 IS A CORE COMPONENT OF THE RBX E 575 00:24:04,840 --> 00:24:07,880 3 UBIQUITIN LIGASE COMPLEX AND 576 00:24:07,880 --> 00:24:10,240 DDB 2 RECRUITS THAT DAMAGED 577 00:24:10,240 --> 00:24:12,280 CHROMATIN. 578 00:24:12,280 --> 00:24:16,520 A MODEL HERE, IT RECRUITS 579 00:24:16,520 --> 00:24:19,400 DAMAGED CHROMATIN AND USES 580 00:24:19,400 --> 00:24:21,760 UBIQUITINATION H 2LA TO MAKE IT 581 00:24:21,760 --> 00:24:26,640 MORE ACCESSIBLE FOR REPAIR BUT 582 00:24:26,640 --> 00:24:30,440 ALSO UBIQUITINDDB 120. 583 00:24:30,440 --> 00:24:31,960 A MAJOR QUESTION WE HAD GOING 584 00:24:31,960 --> 00:24:35,280 INTO THIS, DDB 1 AND 2 INTERACT 585 00:24:35,280 --> 00:24:38,720 STRONGLY AND YOU CAN BEURRE PHI 586 00:24:38,720 --> 00:24:40,720 THEM TOGETHER BUT IN CELLS UVDDB 587 00:24:40,720 --> 00:24:43,080 ACTS AS OBLIGATE HETERODIMER OR 588 00:24:43,080 --> 00:24:44,360 IS THERE MORE DYNAMICS AT PLAY 589 00:24:44,360 --> 00:24:46,680 IN THE SYSTEM. 590 00:24:46,680 --> 00:24:49,240 SO TO TEST IT WE LABELED DDB 1 591 00:24:49,240 --> 00:24:53,200 WITH EGFP AND DDB 2 WITH HALO SO 592 00:24:53,200 --> 00:24:54,920 IN RED AND BLUE RESPECTIVELY AND 593 00:24:54,920 --> 00:24:56,960 WHEN BOTH ON THE DNA AT THE SAME 594 00:24:56,960 --> 00:24:58,160 TIME THAT FORM A PINK COLOR 595 00:24:58,160 --> 00:25:00,920 BECAUSE THEY ARE FLUORESCENT 596 00:25:00,920 --> 00:25:02,800 SIMULTANEOUSLY SO WITH THE DNA 597 00:25:02,800 --> 00:25:04,760 WE TREATS WITH UV TO DAMAGE AT 598 00:25:04,760 --> 00:25:05,880 SEVERAL SITES. 599 00:25:05,880 --> 00:25:08,280 SO THIS IS HOW THE CHEMO GRAPHS 600 00:25:08,280 --> 00:25:09,200 COME UP IN THE INSTRUMENT, YOU 601 00:25:09,200 --> 00:25:10,640 CAN SEE THE LINES UP HERE AND 602 00:25:10,640 --> 00:25:12,520 THEY DISAPPEAR. 603 00:25:12,520 --> 00:25:14,920 THAT'S VISITING THE DNA AND 604 00:25:14,920 --> 00:25:15,680 ASSOCIATE OF THE DNA. 605 00:25:15,680 --> 00:25:18,240 YOU CAN SEE IT IS NOT ALL PINK 606 00:25:18,240 --> 00:25:19,920 LINES, WE HAVE RED LINES WE HAVE 607 00:25:19,920 --> 00:25:24,480 BLUE LINES AND COMBINATION. 608 00:25:24,480 --> 00:25:26,680 SO IMPORTANTLY THE POSITIONS 609 00:25:26,680 --> 00:25:29,240 VISITED MULTIPLE TIMES 610 00:25:29,240 --> 00:25:30,720 CONSISTENT WITH MAYBE REVISITING 611 00:25:30,720 --> 00:25:32,720 THE SAME SITE OF DAMAGE OVER AND 612 00:25:32,720 --> 00:25:35,240 OVER AGAIN, THERE IS ALSO SOME 613 00:25:35,240 --> 00:25:37,400 LIKE THIS WHERE THE COMPLEX 614 00:25:37,400 --> 00:25:39,720 COMES IN TOGETHER AND DDB 1 615 00:25:39,720 --> 00:25:43,080 DISSOCIATES AND DDB 2. 616 00:25:43,080 --> 00:25:46,640 SODS WHAT WE CAN DO IS FIT THESE 617 00:25:46,640 --> 00:25:49,960 FLUORESCENT INTENSITY OF THESE 618 00:25:49,960 --> 00:25:52,000 BINDING EVENTS TO CRTD WHICH IS 619 00:25:52,000 --> 00:25:54,960 A CUMULATIVE RESIDENCE 620 00:25:54,960 --> 00:25:55,720 DISTRIBUTION BUT YOU CAN THINK 621 00:25:55,720 --> 00:25:58,400 OF THIS AS A SURVIVAL CURVE SO 622 00:25:58,400 --> 00:26:00,960 EACH EVENT SHORT ONES IN ONE 623 00:26:00,960 --> 00:26:02,560 SECOND GO TO LEFT SIDE, LONG 624 00:26:02,560 --> 00:26:04,080 ONES THAT ARE 100 SECONDS OR 625 00:26:04,080 --> 00:26:08,480 MORE GO TO THIS SIDE. 626 00:26:08,480 --> 00:26:11,080 WHEN WE SORT WE CAN FIT EXKNOW 627 00:26:11,080 --> 00:26:13,840 ANYONIAL. 628 00:26:13,840 --> 00:26:16,840 FIT THIS TRIPLE EXPONENTIAL K 629 00:26:16,840 --> 00:26:18,760 FUNCTION WITH WEIGHTED TAU 630 00:26:18,760 --> 00:26:19,840 AVERAGE 29 SECONDS FOR BOTH. 631 00:26:19,840 --> 00:26:22,040 THIS IS CONSISTENT WITH WHAT IS 632 00:26:22,040 --> 00:26:27,040 OBSERVED WITH PURIFIED PROTEIN. 633 00:26:27,040 --> 00:26:30,000 IMPORTANTLY WHEN LOOK AT 634 00:26:30,000 --> 00:26:32,320 CO-LOCALIZATION, 32% LOCALIZED 635 00:26:32,320 --> 00:26:34,640 WITH 30 AND 38% RESPECTIVELY, 636 00:26:34,640 --> 00:26:36,880 INDEPENDENT. 637 00:26:36,880 --> 00:26:39,680 WE CAN BREAK DOWN CO-LOCALIZE TO 638 00:26:39,680 --> 00:26:41,560 SEE WHICH GOT THERE FIRST AND 639 00:26:41,560 --> 00:26:43,200 WHICH LEFT FIRST SO WHEN YOU 640 00:26:43,200 --> 00:26:44,920 BREAK DOWN TWO COLORS THERE IS 641 00:26:44,920 --> 00:26:47,440 11 EVENTS SO IN THIS CATEGORY 3 642 00:26:47,440 --> 00:26:50,240 FOR INSTANCE DDB 1 SHOWS UP AND 643 00:26:50,240 --> 00:26:53,240 DDB 2 SHOWS UP, DDB 1 LEAVES AND 644 00:26:53,240 --> 00:26:54,560 THEN DDB 2. 645 00:26:54,560 --> 00:26:57,680 THE MODEL CONSISTENT WITH THE 646 00:26:57,680 --> 00:26:59,000 LITERATURE WOULD BE NUMBER 7. 647 00:26:59,000 --> 00:27:02,640 THAT IS THE MOST COMMON CATEGORY 648 00:27:02,640 --> 00:27:03,560 WE SEE. 649 00:27:03,560 --> 00:27:06,120 WE ALSO SEE CATEGORIES WE DIDN'T 650 00:27:06,120 --> 00:27:08,880 EXPECT LIKE NUMBER 9 WHERE DDB 2 651 00:27:08,880 --> 00:27:10,840 GETS THERE FIRST, THEN DDB 1 652 00:27:10,840 --> 00:27:14,440 VISITS AND LEAVES AND THEN DDB 653 00:27:14,440 --> 00:27:15,920 2. 654 00:27:15,920 --> 00:27:19,240 WE RARELY SAW CATEGORY 3, 4, 5 655 00:27:19,240 --> 00:27:22,720 FOR DDB 1 ARRIVES FIRST BECAUSE 656 00:27:22,720 --> 00:27:24,720 IT DOESN'T INTERACT DIRECTLY 657 00:27:24,720 --> 00:27:25,720 WITH THE DNA DAMAGE. 658 00:27:25,720 --> 00:27:27,000 IMPORTANT WE WERE ABLE TO MAKE 659 00:27:27,000 --> 00:27:29,440 MUTATION OF DDB 2 RIGHT INTO 660 00:27:29,440 --> 00:27:31,880 ACTIVE SITE INTERACTS WITH PHOTO 661 00:27:31,880 --> 00:27:33,200 PRODUCTS AND INSTEAD OF BEING 662 00:27:33,200 --> 00:27:35,840 STRAIGHT LINE BECAUSE IT CAN'T 663 00:27:35,840 --> 00:27:37,440 ENGAGE DAMAGE EFFECTIVELY THE 664 00:27:37,440 --> 00:27:38,880 CHEMO GRAPH MOVES ON THE DNA 665 00:27:38,880 --> 00:27:41,920 WHICH IS INDICATIVE OF UVDDB 666 00:27:41,920 --> 00:27:43,760 SPLICING ON THE DDB. 667 00:27:43,760 --> 00:27:45,560 SO INDEED THE LIFETIME IS 668 00:27:45,560 --> 00:27:48,840 SHORTER FOR THE MUTANT FORM OF 669 00:27:48,840 --> 00:27:54,200 DDB 2, K 244E AND IT WAS LOT 670 00:27:54,200 --> 00:27:57,280 GREATER PERCENTAGE OF EVENTS 671 00:27:57,280 --> 00:27:59,240 MODAL MEAN THE SLID ON THE DAB 672 00:27:59,240 --> 00:28:00,520 OR MOVED AFTER BINDING. 673 00:28:00,520 --> 00:28:02,520 I WANT TO TALK A FEW CONTROLS WE 674 00:28:02,520 --> 00:28:03,920 HAVE GONE THROUGH, FIRST WE CAN 675 00:28:03,920 --> 00:28:06,240 MEASURE THE CONCENTRATION OF THE 676 00:28:06,240 --> 00:28:08,320 PROTEIN BY LOOKING AT THE 677 00:28:08,320 --> 00:28:10,640 FLUORESCENCE INTENSITY, HERE ARE 678 00:28:10,640 --> 00:28:11,240 THREE CONCENTRATIONS, YOU CAN 679 00:28:11,240 --> 00:28:14,200 SEE EVENT IN THE PICA MOLAR 680 00:28:14,200 --> 00:28:16,320 SCALE WE CAN SEE BACKGROUND 681 00:28:16,320 --> 00:28:16,960 FLUORESCENCE. 682 00:28:16,960 --> 00:28:19,960 SO GOOD STANDARD CURVE TO 683 00:28:19,960 --> 00:28:21,600 DETERMINE GFP CONCENTRATION. 684 00:28:21,600 --> 00:28:23,480 WE CAN MEASURE PHOTO BLEACHING 685 00:28:23,480 --> 00:28:27,200 BY LOOKING AT MOLECULES BOUND TO 686 00:28:27,200 --> 00:28:29,040 SURFACE AND WATCHING OVER TIME 687 00:28:29,040 --> 00:28:31,120 SO WE SEE A LONG LIFETIME GREAT 688 00:28:31,120 --> 00:28:32,840 FOR THIS APPROACH BECAUSE WE SEE 689 00:28:32,840 --> 00:28:34,960 THESE ALONG WITH EVENTS AND 690 00:28:34,960 --> 00:28:35,760 LOOKING (INAUDIBLE) CAN HELP 691 00:28:35,760 --> 00:28:36,120 WITH THAT. 692 00:28:36,120 --> 00:28:37,920 BY THE WAY WE CAN USE A WESTERN 693 00:28:37,920 --> 00:28:40,040 BLOT TO COMPARE THE ENDOGENOUS 694 00:28:40,040 --> 00:28:41,680 LEVEL TO THE OVEREXPRESS LEVELS 695 00:28:41,680 --> 00:28:43,960 SO THIS IS A TRANSIENT 696 00:28:43,960 --> 00:28:46,120 EXPRESSION IN U2OS CELLS, THIS 697 00:28:46,120 --> 00:28:47,440 IS PURIFIED DDB #. 698 00:28:47,440 --> 00:28:51,000 YOU CAN SEE OVEREXPRESS PROTEIN 699 00:28:51,000 --> 00:28:53,240 OUTCOMPETES, AT MUCH HIGHER 700 00:28:53,240 --> 00:28:54,240 CONCENTRATION THAN ENDOGENOUS 701 00:28:54,240 --> 00:29:00,240 LEVEL. 702 00:29:00,240 --> 00:29:03,240 WITH THAT I WANT TO SAY SMADNE 703 00:29:03,240 --> 00:29:05,040 IS AWESOME TECHNIQUE, WE CAN GO 704 00:29:05,040 --> 00:29:06,120 -- IT IS AN ENVIRONMENTS THAT 705 00:29:06,120 --> 00:29:08,320 HAS THE OTHER PARTNERS THAT ARE 706 00:29:08,320 --> 00:29:10,240 CLOSELY MIMICKING THE KNEW COLOR 707 00:29:10,240 --> 00:29:11,720 ENVIRONMENT AND PRESEVERRING 708 00:29:11,720 --> 00:29:14,080 PROTEIN PTM, NOT JUST DNA REPAIR 709 00:29:14,080 --> 00:29:16,600 PROTEIN, ANY PROTEIN THAT CAN BE 710 00:29:16,600 --> 00:29:18,120 TAGGED CAN BIND TO DNA CAN BE 711 00:29:18,120 --> 00:29:19,720 STUDIED ON THE SYSTEM AND THE 712 00:29:19,720 --> 00:29:22,960 MULTIPLE COLORS REALLY UNVEIL 713 00:29:22,960 --> 00:29:25,160 ORDERS OF ASSEMBLY AND IT IS 714 00:29:25,160 --> 00:29:26,280 ASSEMBLY SO WE HAVE STUDIED A 715 00:29:26,280 --> 00:29:29,600 LOT OF PROTEINS INCLUDING DDB 1, 716 00:29:29,600 --> 00:29:35,200 2, ODG 1 GLYCOSYLATION, PARP1 717 00:29:35,200 --> 00:29:41,280 PARP 2, BETA DDG AND XPC. 718 00:29:41,280 --> 00:29:42,560 THE GOAL GOING FORWARD IS 719 00:29:42,560 --> 00:29:43,760 CONSTITUTE HOW THE PATHWAY WORKS 720 00:29:43,760 --> 00:29:45,240 AT THE SINGLE MOLECULE LEVEL 721 00:29:45,240 --> 00:29:48,240 LOOKING AT TWO OR THREE MULTIPLE 722 00:29:48,240 --> 00:29:48,800 SEGMENTS. 723 00:29:48,800 --> 00:29:50,360 SO WITH THAT I WOULD LIKE TO 724 00:29:50,360 --> 00:29:53,240 ACKNOWLEDGE THE LAB MEMBERS, MY 725 00:29:53,240 --> 00:29:57,120 POSTBACK MENTORSHIP COMMITTEE, 726 00:29:57,120 --> 00:29:59,360 COLLABORACOLLABORATORS. 727 00:29:59,360 --> 00:30:00,480 WITH THAT THANK YOU FOR YOUR 728 00:30:00,480 --> 00:30:01,280 ATTENTION. 729 00:30:01,280 --> 00:30:01,760 >> THANK YOU. 730 00:30:01,760 --> 00:30:07,600 THANK YOU FOR EXCELLENT TALK. 731 00:30:07,600 --> 00:30:12,360 WE WILL GO ON TO BRUCE WHO IS 732 00:30:12,360 --> 00:30:16,360 INTRODUCING THE LAST SPEAKER. 733 00:30:16,360 --> 00:30:16,840 >> STONY BROOK. 734 00:30:16,840 --> 00:30:18,360 >> SO THIS WILL BE SOMETHING OF 735 00:30:18,360 --> 00:30:22,040 A CHANG OF PACE. 736 00:30:22,040 --> 00:30:23,480 THIS THAT YOU MAY HAVE GLEANED 737 00:30:23,480 --> 00:30:24,200 FROM THE TITLE. 738 00:30:24,200 --> 00:30:26,920 JAMIE JOINED THE LAB IN 2018 AS 739 00:30:26,920 --> 00:30:30,320 MASTERS STUDENT, HE CAME FROM 740 00:30:30,320 --> 00:30:30,840 TAIWAN. 741 00:30:30,840 --> 00:30:32,360 HE STARTED WORKING ON THIS 742 00:30:32,360 --> 00:30:34,520 PROJECT STRAIGHT AWAY, FOLLOWING 743 00:30:34,520 --> 00:30:36,000 ON ANOTHER GRADUATE STUDENT'S 744 00:30:36,000 --> 00:30:38,920 WORK HERE. 745 00:30:38,920 --> 00:30:41,400 HE A YEAR LATER TRANSITIONED TO 746 00:30:41,400 --> 00:30:43,280 THE Ph.D. PROGRAM. 747 00:30:43,280 --> 00:30:45,440 SO JUST BEFORE THE PANDEMIC IN 748 00:30:45,440 --> 00:30:48,880 2019 HE STARTED AS A Ph.D. 749 00:30:48,880 --> 00:30:49,480 STUDENT. 750 00:30:49,480 --> 00:30:51,600 WHICH HAD ALL THE SAME EFFECT AS 751 00:30:51,600 --> 00:30:53,400 IT DID ON EVERYONE ELSE, HE GOT 752 00:30:53,400 --> 00:30:54,920 THROUGH THAT, WORKING WHEN WE 753 00:30:54,920 --> 00:30:58,960 GOT BACK TO THE LAB. 754 00:30:58,960 --> 00:31:02,560 HIS PROJECT IS FOCUSED ON 755 00:31:02,560 --> 00:31:04,440 COMPLEX INTERACTIONS, HOW 756 00:31:04,440 --> 00:31:13,920 COMPLEX MATERIALS, IN THIS CASE 757 00:31:13,920 --> 00:31:17,560 SIMULATING OF LIEU MARCH CELLS 758 00:31:17,560 --> 00:31:21,240 CAUSE THE DUST -- LUNAR CELLS. 759 00:31:21,240 --> 00:31:24,400 SO HE HAS A LOT TO SAY, I HOPE 760 00:31:24,400 --> 00:31:25,400 HECK SQUEEZE IT IN IN 15 761 00:31:25,400 --> 00:31:26,200 MINUTES. 762 00:31:26,200 --> 00:31:28,120 GGO AHEAD. 763 00:31:28,120 --> 00:31:33,840 >> HI, EVERYONE, THIS IS JAMIE, 764 00:31:33,840 --> 00:31:35,040 I'M IN THE -- TODAY I WANT TO 765 00:31:35,040 --> 00:31:38,080 TALK ABOUT MY PROJECT, THE 766 00:31:38,080 --> 00:31:42,880 IMPACT OF ARTIFICIALLY SPACE 767 00:31:42,880 --> 00:31:45,640 WEATHERED LUNAR DUST SIMULANTS 768 00:31:45,640 --> 00:31:47,200 IN HUMAN LUNG CELLS. 769 00:31:47,200 --> 00:31:52,920 BACK IN THE 'O '60S WHEN NASA WT 770 00:31:52,920 --> 00:31:55,400 TO UPHOLD A PROGRAM THEY HAVE 771 00:31:55,400 --> 00:31:56,000 LIMITED SIZE ON THE MOON THERE 772 00:31:56,000 --> 00:31:59,760 FOR ANALYSIS, HOWEVER THEY 773 00:31:59,760 --> 00:32:01,760 UNDERESTIMATED OR ALREADY WERE 774 00:32:01,760 --> 00:32:04,240 NOT AWARE OF RISK OF LUNAR DUST. 775 00:32:04,240 --> 00:32:05,880 THE FINAL REPORT IT MENTIONED 776 00:32:05,880 --> 00:32:08,000 THE LUNAR DUST CAN GO EVERYWHERE 777 00:32:08,000 --> 00:32:09,520 AND THE PICTURE HAS INDICATED, 778 00:32:09,520 --> 00:32:12,320 YOU CAN SEE THAT THE LUNAR DUST 779 00:32:12,320 --> 00:32:15,360 AFTER THEIR GOING BACK TO THE 780 00:32:15,360 --> 00:32:16,560 EARTH, BASICALLY IT IS 781 00:32:16,560 --> 00:32:18,440 EVERYWHERE BECAUSE YOU CAN SEE 782 00:32:18,440 --> 00:32:23,320 THE DUST IS COVERING THE FACE OF 783 00:32:23,320 --> 00:32:26,320 THE ASTRONAUTS AND THE SHIRTS. 784 00:32:26,320 --> 00:32:28,440 THOSE MISSION CREW MEMBERS ALSO 785 00:32:28,440 --> 00:32:29,960 MENTIONED THAT AFTER BREATHING 786 00:32:29,960 --> 00:32:35,120 IN THOSE LUNAR DUST PARTICLES 787 00:32:35,120 --> 00:32:36,440 THEY FELL RUNNY NOSES. 788 00:32:36,440 --> 00:32:38,120 SO NASA WANTS TO KNOW WHAT 789 00:32:38,120 --> 00:32:39,840 EXACTLY IS HAPPENING WITH THOSE 790 00:32:39,840 --> 00:32:43,360 DUST AND THEY DID SOME ANALYSIS 791 00:32:43,360 --> 00:32:46,040 UNDER THE COMPOSITION AND 792 00:32:46,040 --> 00:32:47,680 CATEGORIZED THEM BY ARRAY THE 793 00:32:47,680 --> 00:32:54,480 LOWER TERRAIN ON THE MOON AND 794 00:32:54,480 --> 00:32:57,280 NOW IN THE PROGRAM ANNOUNCED IN 795 00:32:57,280 --> 00:32:59,200 2017, HAIR GOAL IS TO BRING 796 00:32:59,200 --> 00:33:03,480 HUMANS BACK TO MOON AND ALSO 797 00:33:03,480 --> 00:33:05,480 EXPLORE NEIGHBORING PLANET MARS. 798 00:33:05,480 --> 00:33:07,560 HOWEVER AS I SAID, POTENTIAL 799 00:33:07,560 --> 00:33:09,120 RISK OF CHALLENGING PART OF THE 800 00:33:09,120 --> 00:33:11,240 LUNAR DUST HAS NOT BEEN OVERCOME 801 00:33:11,240 --> 00:33:11,880 YET. 802 00:33:11,880 --> 00:33:13,880 AND SO THAT IS WHY TODAY I'M 803 00:33:13,880 --> 00:33:16,560 GOING TO GIVE THE BROADER 804 00:33:16,560 --> 00:33:18,400 INTRODUCTION TO YOU TO ELABORATE 805 00:33:18,400 --> 00:33:20,920 ON WHY IT IS NOT OVERCOME YET. 806 00:33:20,920 --> 00:33:24,560 SO FIRST AFTER THEIR ANALYSIS ON 807 00:33:24,560 --> 00:33:25,880 LUNAR DUST THEY HAVE FOUND OUT 808 00:33:25,880 --> 00:33:31,680 DUST IS SIMILAR TO TERRESTRIAL 809 00:33:31,680 --> 00:33:31,880 DUST. 810 00:33:31,880 --> 00:33:35,760 HOWEVER CHRONIC EXPOSURE TO 811 00:33:35,760 --> 00:33:38,640 LUNAR DUST TERRESTRIAL DUST CAN 812 00:33:38,640 --> 00:33:41,880 LEAD TO (INAUDIBLE) SO WITHOUT 813 00:33:41,880 --> 00:33:44,880 THE ATMOSPHERE -- WITH THAT 814 00:33:44,880 --> 00:33:47,600 PERCEPTION OF ATMOSPHERE ON THE 815 00:33:47,600 --> 00:33:50,720 MOON THIS COULD BE WAY MORE 816 00:33:50,720 --> 00:33:51,800 TOXIC ENVIRONMENT THAN WHEN 817 00:33:51,800 --> 00:33:56,560 COMPARING TO EXPOSURE TO 818 00:33:56,560 --> 00:33:57,160 (INAUDIBLE) DUST. 819 00:33:57,160 --> 00:33:59,040 THOSE IN FACT NOT CHAPPING ON 820 00:33:59,040 --> 00:34:02,560 EARTH, IT IS EXCLUSIVE ON MOON, 821 00:34:02,560 --> 00:34:06,200 INCLUDING THE BOMBARDMENT OF 822 00:34:06,200 --> 00:34:06,840 MICROMY YOUR RIOTS WHICH MAKESES 823 00:34:06,840 --> 00:34:10,280 THE DUST FINER AND ALSO SOLAR 824 00:34:10,280 --> 00:34:14,920 WHEN BLOWING SOLAR WIND WHICH 825 00:34:14,920 --> 00:34:19,920 CONTAINS A LOT OF LUNAR DUST AND 826 00:34:19,920 --> 00:34:22,040 UVY ALSO LEADS TO CHANG PROPERTY 827 00:34:22,040 --> 00:34:24,240 PHYSICALLY AND CHEMICALLY TO THE 828 00:34:24,240 --> 00:34:24,760 LUNAR DUST. 829 00:34:24,760 --> 00:34:26,960 THAT IS WHY THE DUST IS MORE 830 00:34:26,960 --> 00:34:28,280 IRRITATING AND COULD STICK TO 831 00:34:28,280 --> 00:34:31,520 EVERYTHING AND GET EVERYWHERE 832 00:34:31,520 --> 00:34:34,480 AFTER LIKE THOSE ASTRONAUTS HAVE 833 00:34:34,480 --> 00:34:37,760 PASSED THROUGH IT. 834 00:34:37,760 --> 00:34:41,160 WITH THE GOAL OF 835 00:34:41,160 --> 00:34:44,240 LOOKING AT EXPOSURE ON THE MOON 836 00:34:44,240 --> 00:34:46,560 WE HYPOTHESIZE OCCUPATIONAL 837 00:34:46,560 --> 00:34:49,680 EXPOSURE LEAD TO DNA MUTATION IN 838 00:34:49,680 --> 00:34:52,080 CELLS AND CAN LEAD TO LONG TERM 839 00:34:52,080 --> 00:34:53,920 INFLAMMATION AND MITOCHONDRIAL 840 00:34:53,920 --> 00:34:55,960 DYSFUNCTION AFTER EXPOSURE BASED 841 00:34:55,960 --> 00:34:58,320 ON PUBLISHED PAPER. 842 00:34:58,320 --> 00:35:02,240 SUCH ATTENUATION OF THOSE 843 00:35:02,240 --> 00:35:04,160 DETRIMENTAL AFFECT LEAD TO 844 00:35:04,160 --> 00:35:07,760 TISSUE DAMAGE IN LUNG, OR OTHER 845 00:35:07,760 --> 00:35:10,240 TISSUE AND IT COULD ELEVATE THE 846 00:35:10,240 --> 00:35:13,920 RISK OF CHRONIC DISEASE AND EVEN 847 00:35:13,920 --> 00:35:14,480 CANCER. 848 00:35:14,480 --> 00:35:16,840 SO YOU MAY BE ASKING OH, WHERE 849 00:35:16,840 --> 00:35:18,760 ARE WE GOING TO GET LUNAR DUST? 850 00:35:18,760 --> 00:35:22,520 WE ARE NOT USING THE LUNAR DUST 851 00:35:22,520 --> 00:35:24,080 BECAUSE THE OTHER -- THERE IS 852 00:35:24,080 --> 00:35:25,960 NOT A LOT OF LUNAR DUST COLLECT 853 00:35:25,960 --> 00:35:26,760 MISDEMEANOR THE PREVIOUS MISSION 854 00:35:26,760 --> 00:35:28,360 AND THE SECOND POINT, IT'S BEEN 855 00:35:28,360 --> 00:35:30,360 50 YEARS. 856 00:35:30,360 --> 00:35:32,560 SO THE CONDITION -- LUNAR DUST 857 00:35:32,560 --> 00:35:34,320 IS NOT THAT (INAUDIBLE) ANY MORE 858 00:35:34,320 --> 00:35:37,160 SO WE ARE TRYING TO SIMULATE 859 00:35:37,160 --> 00:35:38,560 LUNAR ENVIRONMENT OURSELF AND 860 00:35:38,560 --> 00:35:47,520 GET THE LUNAR DUST SIMULANTS 861 00:35:47,520 --> 00:35:49,400 WHICH SIMULATES THE COMPOSITION 862 00:35:49,400 --> 00:35:51,080 OF MARE. 863 00:35:51,080 --> 00:35:57,680 THE OTHER LUNAR SIMULANT 864 00:35:57,680 --> 00:35:59,720 COMPOSITION ON THE HIGH LANDS ON 865 00:35:59,720 --> 00:36:00,080 THE MOON. 866 00:36:00,080 --> 00:36:01,960 WE BOUGHT IT FROM A MANUFACTURER 867 00:36:01,960 --> 00:36:05,640 AND COLLABORATOR WOULD DO A 868 00:36:05,640 --> 00:36:06,800 OCCLUSION FOR THOSE PARTICLES SO 869 00:36:06,800 --> 00:36:11,600 WE CAN MAKE SURE THAT THE DUST 870 00:36:11,600 --> 00:36:15,000 PARTICLES ARE BELOW TEN MICRONS, 871 00:36:15,000 --> 00:36:18,280 SO THE SIGNIFICANCE OF THOSE 872 00:36:18,280 --> 00:36:24,120 FINE PARTICLES WITH THE -- AFTER 873 00:36:24,120 --> 00:36:26,640 -- ASTRONAUT LEADS TO DIFFERENT 874 00:36:26,640 --> 00:36:28,840 (INAUDIBLE) OR RISKS AFTER SUCH 875 00:36:28,840 --> 00:36:33,520 EXPOSURE. 876 00:36:33,520 --> 00:36:35,600 THEN NEXT AFTER GETTING THOSE 877 00:36:35,600 --> 00:36:40,760 FINE PARTICLES WE WOULD SORT 878 00:36:40,760 --> 00:36:44,280 INTO (INAUDIBLE) ENSURE 879 00:36:44,280 --> 00:36:45,600 FRESHNESS AND AS ANOTHER PART TO 880 00:36:45,600 --> 00:36:48,080 MENTION IS THAT I MENTION 881 00:36:48,080 --> 00:36:51,640 EARLIER IT IS A PROCESS THAT 882 00:36:51,640 --> 00:36:52,880 EXCLUSIVELY HAPPEN ON MOON. 883 00:36:52,880 --> 00:36:58,200 HERE WE REDUCE THE DUST 884 00:36:58,200 --> 00:37:00,200 PARTICLES TO SOLAR WIND WHICH IS 885 00:37:00,200 --> 00:37:03,680 AN ASSET I MENTIONED EARLIER OF 886 00:37:03,680 --> 00:37:04,280 SPACE (INAUDIBLE) THAT IS WHY 887 00:37:04,280 --> 00:37:06,960 THE TITLE I'M TALKING ABOUT 888 00:37:06,960 --> 00:37:07,600 ARTIFICIAL SPATE ARTERY BECAUSE 889 00:37:07,600 --> 00:37:08,800 WE ARE TRYING TO DO THAT, 890 00:37:08,800 --> 00:37:10,920 SIMULATE THIS IN OUR 891 00:37:10,920 --> 00:37:13,400 COLLABORATORS LAB. 892 00:37:13,400 --> 00:37:20,160 AFTER WE WE WILL GRIND OFF 893 00:37:20,160 --> 00:37:22,000 SPECIFIC TO LUNG EPITHELIAL 894 00:37:22,000 --> 00:37:26,480 CELLS AS OUR MODEL HERE. 895 00:37:26,480 --> 00:37:27,800 FIRST EXPERIMENT TALK ABOUT FOR 896 00:37:27,800 --> 00:37:32,400 TESTING OF TOXICITY OF LUNAR 897 00:37:32,400 --> 00:37:35,280 DUST IS (INAUDIBLE) A DYE THAT 898 00:37:35,280 --> 00:37:39,800 IS OXIDIZED BY SUPER OX SIDE 899 00:37:39,800 --> 00:37:41,240 INTO THE MITOCHONDRIA. 900 00:37:41,240 --> 00:37:45,400 SO WHEN THEY ARE OVERLOAD 901 00:37:45,400 --> 00:37:48,000 OVERLOADED OUTSIDE THE 902 00:37:48,000 --> 00:37:50,440 MITOCHONDRIA, THE ELECTRON CHAIN 903 00:37:50,440 --> 00:37:59,640 OR OUTSI OUTER SOURCES THESE SUB 904 00:37:59,640 --> 00:38:01,120 COMPLEX WOULD EMIT OF 905 00:38:01,120 --> 00:38:04,200 QUANTIFICATION, WE MEASURE 906 00:38:04,200 --> 00:38:06,480 SIGNAL OF THE RESIN TO SEE HOW 907 00:38:06,480 --> 00:38:08,760 MUCH IS INSIDE MITOCHONDRIA 908 00:38:08,760 --> 00:38:11,440 AFTER EXPOSURE TO LUNAR DUST 909 00:38:11,440 --> 00:38:14,680 SIMULANTS SO WE CAN DEVELOP THE 910 00:38:14,680 --> 00:38:16,160 MITOCHONDRIA FUNCTION. 911 00:38:16,160 --> 00:38:18,200 BEFORE I TALK FUNCTION THERE IS 912 00:38:18,200 --> 00:38:20,560 OVERVIEW TO SHOW EVERYONE. 913 00:38:20,560 --> 00:38:24,000 THIS IS THE CELL SURVIVAL AFTER 914 00:38:24,000 --> 00:38:27,000 EXPOSURE TO 1 TO 2 THAT I 915 00:38:27,000 --> 00:38:27,920 MENTIONED EARLIER. 916 00:38:27,920 --> 00:38:31,000 AFTER REDUCTION OF THOSE TWO 917 00:38:31,000 --> 00:38:36,600 LUNAR DUST EXPOSURE OF MATERIALS 918 00:38:36,600 --> 00:38:38,800 LEAD TO MORE CELL DEATH, 919 00:38:38,800 --> 00:38:41,440 UNTREATED ONES SURVIVAL IS 920 00:38:41,440 --> 00:38:44,080 AROUND 50 TO 60 BUT AT THE 921 00:38:44,080 --> 00:38:48,200 CONCENTRATION OF .5 MILLIGRAMS, 922 00:38:48,200 --> 00:38:50,520 HOWEVER IF WE TREAT EXPOE THE 923 00:38:50,520 --> 00:38:52,840 CELLS REDUCE ONE YOU CAN SEE FAR 924 00:38:52,840 --> 00:38:56,800 WAY LOWER THAN THE UNTREATED 925 00:38:56,800 --> 00:38:58,560 ONE. 926 00:38:58,560 --> 00:38:59,640 IN MITOCHONDRIAL FUNCTION YOU 927 00:38:59,640 --> 00:39:02,320 CAN SEE IN TREATED, THERE IS 928 00:39:02,320 --> 00:39:03,680 ALMOST NO SIGNAL REPORTED. 929 00:39:03,680 --> 00:39:09,440 HOWEVER AFTER EXPOSURE TO 930 00:39:09,440 --> 00:39:10,320 SIMULANTS YOU CAN SEE THE 931 00:39:10,320 --> 00:39:12,640 MITOCHONDRIAL COMPLEX HAS BEEN 932 00:39:12,640 --> 00:39:14,520 GAINED WITH (INAUDIBLE) AND 933 00:39:14,520 --> 00:39:16,080 QUANTIFY THOSE IMAGES AND FOUND 934 00:39:16,080 --> 00:39:18,200 THE SAME PHENOMENON AFTER 935 00:39:18,200 --> 00:39:20,920 EXPOSURE TO LUNAR DUST 936 00:39:20,920 --> 00:39:21,960 SIMULANTS, THOUGH SIGNAL ARE 937 00:39:21,960 --> 00:39:24,080 SIGNIFICANTLY INCREASED THE 938 00:39:24,080 --> 00:39:26,400 REDUCE WILL SHOW MUCH HIGHER 939 00:39:26,400 --> 00:39:28,080 LEVEL OF THE (INAUDIBLE) WHICH 940 00:39:28,080 --> 00:39:33,160 MEANS UNDERGOING MORE INTENTION 941 00:39:33,160 --> 00:39:33,760 MITOCHONDRIAL DYSFUNCTION 942 00:39:33,760 --> 00:39:35,200 SITUATION AFTER EXPOSURE TO 943 00:39:35,200 --> 00:39:36,000 THOSE MATERIALS. 944 00:39:36,000 --> 00:39:38,520 SO AFTER UNDERSTANDING CELL 945 00:39:38,520 --> 00:39:41,200 SURVIVAL AND MITOCHONDRIAL 946 00:39:41,200 --> 00:39:41,680 FUNCTION. 947 00:39:41,680 --> 00:39:44,680 WE LIKE TO SEE MITOCHONDRIAL 948 00:39:44,680 --> 00:39:47,000 INTEGRITY OF MITOCHONDRIAL DNA, 949 00:39:47,000 --> 00:39:50,960 WHICH IS CRUCIAL FOR FUNCTION. 950 00:39:50,960 --> 00:39:53,600 WE USE TWO SETS OF PCR WITH 951 00:39:53,600 --> 00:39:55,640 MECHANISM OF THE PCR ASSAY, SO 952 00:39:55,640 --> 00:39:58,520 WE WILL HAVE THE SHORT PCR AS 953 00:39:58,520 --> 00:40:05,720 CONTROL AND WE WILL RUN PCR 954 00:40:05,720 --> 00:40:07,720 BECAUSE THERE ARE MORE EVENLY 955 00:40:07,720 --> 00:40:10,640 DAMAGED BY THOSE LUNAR DUST 956 00:40:10,640 --> 00:40:11,800 SIMULANTS OR OTHER TOX CAN'T. 957 00:40:11,800 --> 00:40:15,000 AFTER RUNNING PCR WE WILL 958 00:40:15,000 --> 00:40:17,360 QUANTIFY AND THEN NORMALIZE THE 959 00:40:17,360 --> 00:40:17,720 RATIO. 960 00:40:17,720 --> 00:40:19,320 WE CAN ALSO WE ALSO FOUND OUT 961 00:40:19,320 --> 00:40:23,200 THAT AFTER EXPOSURE TO BOTH 962 00:40:23,200 --> 00:40:24,320 SIMULANTS, REDUCE ONE ARE ALSO 963 00:40:24,320 --> 00:40:26,680 SHOWING FAR WAY LOWER INTEGRITY 964 00:40:26,680 --> 00:40:31,080 COMPARED TO THE NON-REDUCE ONES. 965 00:40:31,080 --> 00:40:36,640 LASTLY BUZZ MITOCHONDRIAL 966 00:40:36,640 --> 00:40:38,880 FUNCTION IS DECIDED BY FUNCTION 967 00:40:38,880 --> 00:40:40,760 MITOCHONDRIAL PROTEINS THAT ARE 968 00:40:40,760 --> 00:40:42,680 CODED WHILE REALLY FUNCTIONING 969 00:40:42,680 --> 00:40:45,880 AS THEY SHOULD BE, SO THOSE 970 00:40:45,880 --> 00:40:48,400 PROTEINS ARE ENCODED BY THE 971 00:40:48,400 --> 00:40:48,880 NUCLEAR DNA. 972 00:40:48,880 --> 00:40:53,280 SO WE ALSO USE THAT ASSAY TO RUN 973 00:40:53,280 --> 00:40:55,480 -- WE ALSO RUN SAY SAY TO 974 00:40:55,480 --> 00:40:56,880 EVALUATE NEED FOR DNA DAMAGE. 975 00:40:56,880 --> 00:41:01,000 SO THIS ASSAY INVOLVES THE CELLS 976 00:41:01,000 --> 00:41:05,520 AND THEN LYSE THE CELLS WITH 977 00:41:05,520 --> 00:41:06,240 ELECTROFREESIS. 978 00:41:06,240 --> 00:41:07,920 YOU CAN SEE THE CELL FORM BUG IF 979 00:41:07,920 --> 00:41:11,200 NOT CONTROLS ARE SHOWING WHAT IS 980 00:41:11,200 --> 00:41:13,600 REALLY NICE BROWN FLUORESCENCE. 981 00:41:13,600 --> 00:41:15,880 AFTER QUANTIFYING RESULTS WE 982 00:41:15,880 --> 00:41:17,080 FOUND OUT AFTER EXPOSURE TO 983 00:41:17,080 --> 00:41:21,400 REDUCE SAMPLES SHOWN HIGHER 984 00:41:21,400 --> 00:41:26,640 TOXICITY BUT EXAMPLE SHOWING 985 00:41:26,640 --> 00:41:30,680 SIGNIFICANT DAMAGE IN DNA YOU 986 00:41:30,680 --> 00:41:34,160 CAN SEE REDUCE MATERIALS SHOWING 987 00:41:34,160 --> 00:41:39,200 MUCH HIGHER DAMAGE. 988 00:41:39,200 --> 00:41:42,000 FROM THE RESULTS HAVE SHOWN YOU, 989 00:41:42,000 --> 00:41:44,960 WE CAN CONCLUDE LUNAR DUST 990 00:41:44,960 --> 00:41:45,880 SIMULANTS IS TOXIC BUT WHY THEY 991 00:41:45,880 --> 00:41:47,600 ARE TOXIC IS STILL KIND OF NOT 992 00:41:47,600 --> 00:41:50,400 CLEAR. 993 00:41:50,400 --> 00:41:53,600 THERE IS A REVIEW PAPER STATING 994 00:41:53,600 --> 00:41:57,440 LUNAR DUST COULD LEAD TO 995 00:41:57,440 --> 00:42:00,000 PRODUCTION OF PARTICLES WHICH 996 00:42:00,000 --> 00:42:05,080 REPRESENTED BY DILUTION OF THE 997 00:42:05,080 --> 00:42:07,000 MATERIALS DISSOLVED IN SOLUTION. 998 00:42:07,000 --> 00:42:10,720 SOME OF THEM ARE DERIVED BY CELL 999 00:42:10,720 --> 00:42:14,280 SO WHEN THEY THOSE PARTICLES, IT 1000 00:42:14,280 --> 00:42:19,120 SHOULD BE KNOW GENERATE SUCH 1001 00:42:19,120 --> 00:42:22,400 COULD LEAD TO CYTOTOXICITY AND 1002 00:42:22,400 --> 00:42:23,880 GENO TOXICITY I MENTIONED 1003 00:42:23,880 --> 00:42:25,920 EARLIER, ACCUMULATION OF THOSE 1004 00:42:25,920 --> 00:42:30,720 ARE REPAIR DNA LEAD TO CANCER. 1005 00:42:30,720 --> 00:42:35,880 TO VALIDATE WE USE WELL KNOWN 1006 00:42:35,880 --> 00:42:39,760 ANTIOXIDANT NAC AND SO WE CAN 1007 00:42:39,760 --> 00:42:45,800 TREAT CELLS WITH NAC AND NAC CAN 1008 00:42:45,800 --> 00:42:50,120 BASICALLY NEUTRALIZE TOXIC -- 1009 00:42:50,120 --> 00:42:52,280 TOXICITY OR OTHER DAMAGE CAUSED 1010 00:42:52,280 --> 00:42:52,880 BY ROS. 1011 00:42:52,880 --> 00:42:54,960 IF THERE'S INCREASE IN CELL 1012 00:42:54,960 --> 00:42:56,480 SURVIVAL OR OTHER BIOMARKERS I 1013 00:42:56,480 --> 00:43:02,200 MENTIONED EARLIER, THAT MEANS -- 1014 00:43:02,200 --> 00:43:04,480 HAS PEAK TOXICITY OF THE 1015 00:43:04,480 --> 00:43:10,400 (INAUDIBLE). 1016 00:43:10,400 --> 00:43:14,480 AS THE SURVIVAL CELL SURVIVAL 1017 00:43:14,480 --> 00:43:17,200 DATA SHOWN HERE, AFTER THE 1018 00:43:17,200 --> 00:43:20,880 PRE-TREATMENT OF NAC, EXPOSURE 1019 00:43:20,880 --> 00:43:24,080 TO LUNAR DUST SHOWS HIGHER CELL 1020 00:43:24,080 --> 00:43:28,800 SURVIVAL RATE NO MATTER IN 1021 00:43:28,800 --> 00:43:34,680 REDUCE OR NON-REDUCE EXPERIM 1022 00:43:34,680 --> 00:43:35,320 EXPERIMENTAL GROUP. 1023 00:43:35,320 --> 00:43:37,440 FOR THE MITOCHONDRIAL DNA WE 1024 00:43:37,440 --> 00:43:39,600 HAVE SEEN INCREASE IN 1025 00:43:39,600 --> 00:43:41,840 MITOCHONDRIAL DNA INTEGRITY. 1026 00:43:41,840 --> 00:43:43,280 WHICH NOT SHOWN HERE IS THE 1027 00:43:43,280 --> 00:43:45,640 MITOCHONDRIAL FUNCTION IS ALSO 1028 00:43:45,640 --> 00:43:48,080 INCREASED AFTER THE 1029 00:43:48,080 --> 00:43:51,080 PRE-TREATMENT OF THE NAC 1030 00:43:51,080 --> 00:43:52,960 ANTIOXIDANTS SO YOU CAN SEE HERE 1031 00:43:52,960 --> 00:43:55,120 BOTH MARKERS ARE INCREASED AFTER 1032 00:43:55,120 --> 00:43:58,960 THE PRE-TREATMENT OF NAC. 1033 00:43:58,960 --> 00:44:02,280 WE CAN CONCLUDE THAT ALTHOUGH 1034 00:44:02,280 --> 00:44:06,840 LUNAR DUST AFTER EXPOSURE TO 1035 00:44:06,840 --> 00:44:09,280 LUNAR DUST SIMULANTS, WE DID SEE 1036 00:44:09,280 --> 00:44:11,960 INCREASE IN CELL TOXICITY, 1037 00:44:11,960 --> 00:44:15,040 MITOCHONDRIAL DYSFUNCTION AND 1038 00:44:15,040 --> 00:44:17,480 REDUCTION OF BOTH MATERIALS 1039 00:44:17,480 --> 00:44:21,320 WHICH IS THE PART ARTIFICIAL, 1040 00:44:21,320 --> 00:44:23,520 INCREASED TOXICITY TOO. 1041 00:44:23,520 --> 00:44:25,240 BUT WE ALSO HAVE FOUND -- WE 1042 00:44:25,240 --> 00:44:28,680 ALSO HAVE VALIDATED (INAUDIBLE) 1043 00:44:28,680 --> 00:44:31,880 I MENTION EARLIER, KEY TO THE 1044 00:44:31,880 --> 00:44:34,680 TOXICITY OF LUNAR DUST 1045 00:44:34,680 --> 00:44:36,560 SIMULANTS, IN THOSE CELLS. 1046 00:44:36,560 --> 00:44:38,480 SO NOW WE HAVE ELEVATED THAT AND 1047 00:44:38,480 --> 00:44:44,520 WANT TO KNOW IF NAC OXIDASE IS 1048 00:44:44,520 --> 00:44:47,520 KEY TO THE INCREASE PRODUCTION 1049 00:44:47,520 --> 00:44:48,760 OF ROS. 1050 00:44:48,760 --> 00:44:50,360 THAT WILL BE THE NEXT STEP WE 1051 00:44:50,360 --> 00:44:53,880 WILL BE TRYING TO DO. 1052 00:44:53,880 --> 00:44:58,160 WITH INCREASE OF NOX LEADS TO 1053 00:44:58,160 --> 00:44:59,320 DOWNSTREAM BETTER RESPONSE WE 1054 00:44:59,320 --> 00:45:02,520 HAVEN'T LOOK INTO YET SO WE HAVE 1055 00:45:02,520 --> 00:45:06,320 THINGS TO DO IN THE FUTURE. 1056 00:45:06,320 --> 00:45:09,640 I WANT TO THANK EVERYBODY IN MY 1057 00:45:09,640 --> 00:45:13,600 LAB, AND ESPECIALLY THE 1058 00:45:13,600 --> 00:45:17,280 PROVISION OF (INAUDIBLE) 1059 00:45:17,280 --> 00:45:18,440 SPECIFIC COMMITTEE AND OUR 1060 00:45:18,440 --> 00:45:19,560 FUNDING SOURCE AND ALSO ALL 1061 00:45:19,560 --> 00:45:21,080 THESE PEOPLE IN OUR PROGRAM AND 1062 00:45:21,080 --> 00:45:23,520 OUR COLLABORATORS HELPING OUT 1063 00:45:23,520 --> 00:45:26,440 WITH MY EXPERIMENTS. 1064 00:45:26,440 --> 00:45:28,800 THANK YOU FOR YOUR ATTENTION 1065 00:45:28,800 --> 00:45:30,480 TOO. 1066 00:45:30,480 --> 00:45:36,080 LASTLY, MY PI IS HOLDING A 1067 00:45:36,080 --> 00:45:37,560 CONFERENCE ANT DNA DAMAGE AND 1068 00:45:37,560 --> 00:45:40,720 REPAIR IN NOVEMBER 6 AND 9, SO 1069 00:45:40,720 --> 00:45:44,280 FEEL FREE TO REGISTER NOW. 1070 00:45:44,280 --> 00:45:46,840 WITH THE QR CODE I JUST TESTED 1071 00:45:46,840 --> 00:45:48,960 IT AND IT IS WORKING OR GET THE 1072 00:45:48,960 --> 00:45:50,920 LINK FOR THE WEBSITE, IT IS 1073 00:45:50,920 --> 00:45:55,480 ABOUT DNA REPAIR DAMAGE AND 1074 00:45:55,480 --> 00:45:59,800 CANCER FOR AGING AND OTHER 1075 00:45:59,800 --> 00:46:00,160 DISEASES. 1076 00:46:00,160 --> 00:46:01,280 THANK YOU FOR YOUR ATTENTION. 1077 00:46:01,280 --> 00:46:04,560 >> THANK YOU FOR THE VERY 1078 00:46:04,560 --> 00:46:05,920 INTERESTING TALK. 1079 00:46:05,920 --> 00:46:08,200 JAMIE, MATT AND DR. LEE, PLEASE 1080 00:46:08,200 --> 00:46:09,880 UNMUTE, TURN ON YOUR VIDEO SO 1081 00:46:09,880 --> 00:46:13,400 YOU CAN SEE THEM. 1082 00:46:13,400 --> 00:46:18,360 WE WILL DO THE QUESTIONS. 1083 00:46:18,360 --> 00:46:19,600 ACTUALLY (INAUDIBLE) HAS A 1084 00:46:19,600 --> 00:46:20,000 QUESTION. 1085 00:46:20,000 --> 00:46:23,280 YOU CAN JUST ASK YOUR QUESTION. 1086 00:46:23,280 --> 00:46:24,120 >> THANKS. 1087 00:46:24,120 --> 00:46:26,200 THIS QUESTION IS TO JAMIE. 1088 00:46:26,200 --> 00:46:29,280 THANK YOU FOR VERY INTERESTING 1089 00:46:29,280 --> 00:46:29,880 TALK. 1090 00:46:29,880 --> 00:46:31,560 WHAT IS CHEMICAL COMPOSITION FOR 1091 00:46:31,560 --> 00:46:34,080 THE LUNAR DUST? 1092 00:46:34,080 --> 00:46:35,360 HOW MUCH IT IS DIFFERENT FROM 1093 00:46:35,360 --> 00:46:42,400 THE DUST ON THE EARTH? 1094 00:46:42,400 --> 00:46:43,000 A. DID REALLY GOOD QUESTION. 1095 00:46:43,000 --> 00:46:44,280 I'LL ANSWER THE SECOND ONE FIRST 1096 00:46:44,280 --> 00:46:45,120 BECAUSE IT IS EASY. 1097 00:46:45,120 --> 00:46:46,440 HOW MUCH IS IT DIFFERENT FROM 1098 00:46:46,440 --> 00:46:48,760 THE DUST IN THE EARTH. 1099 00:46:48,760 --> 00:46:50,960 IT IS ACTUALLY LIKE COMPOSITION 1100 00:46:50,960 --> 00:46:52,320 WISE, IT IS THE SAME. 1101 00:46:52,320 --> 00:46:54,000 SO THEY ALSO HAVE MAGNESIUM, 1102 00:46:54,000 --> 00:47:01,600 THEY ALSO HAVE IRON, THEY HAVE 1103 00:47:01,600 --> 00:47:04,480 ALMOST EVERY KIND OF ELEMENT, 1104 00:47:04,480 --> 00:47:05,720 CHEMICAL ELEMENT THAT IS PRESENT 1105 00:47:05,720 --> 00:47:07,240 ON EARTH SO BASICALLY THE SAME. 1106 00:47:07,240 --> 00:47:12,280 AS I SAID EARLIER IN THE TALK, 1107 00:47:12,280 --> 00:47:13,120 MANY DIFFERENCES, ATMOSPHERE ON 1108 00:47:13,120 --> 00:47:16,840 THE MOON SO THE MOON WILL 1109 00:47:16,840 --> 00:47:18,320 (INAUDIBLE) CHANG THE PROPERTIES 1110 00:47:18,320 --> 00:47:21,080 SO IT IS PRETTY MUCH THE SAME 1111 00:47:21,080 --> 00:47:23,160 CHEMICAL LINES AND YOU SAID WHAT 1112 00:47:23,160 --> 00:47:27,720 IS THE CHEMICAL COMPOSITION OF 1113 00:47:27,720 --> 00:47:31,640 LUNAR DUST SO TO LEARN THE 1114 00:47:31,640 --> 00:47:33,080 SIMULANT I USE SILICONE DUST, 1115 00:47:33,080 --> 00:47:36,160 COMPOSED OF SOME IRON OXIDE AND 1116 00:47:36,160 --> 00:47:38,920 MAGNESIUM OXIDE, THAT'S KIND OF 1117 00:47:38,920 --> 00:47:42,680 THE MAIN STUFF IN MY LUNAR DUST 1118 00:47:42,680 --> 00:47:43,560 SIMULANTS. 1119 00:47:43,560 --> 00:47:44,920 BUT IN THE KNOCK OUT REPORT, 1120 00:47:44,920 --> 00:47:46,320 THOUGH THE -- THE KNOCK OUT 1121 00:47:46,320 --> 00:47:49,560 REPORT IS ALSO SHOWING SOME 1122 00:47:49,560 --> 00:47:51,520 SIMILAR COMPOSITION, IN TERMS OF 1123 00:47:51,520 --> 00:47:53,640 LIKE -- IN TERMS OF THE 1124 00:47:53,640 --> 00:47:55,240 COMPOSITION, IT IS SIMILAR, NOT 1125 00:47:55,240 --> 00:47:58,800 REALLY FOR THE SILICA DUST, 1126 00:47:58,800 --> 00:48:01,040 SILICA DUST IS KIND OF A COMMON 1127 00:48:01,040 --> 00:48:04,040 MATERIAL PRESENT ON THE MOON AND 1128 00:48:04,040 --> 00:48:04,840 EARTH. 1129 00:48:04,840 --> 00:48:07,280 OTHER DIFFERENT SITES BECAUSE I 1130 00:48:07,280 --> 00:48:09,720 ALSO MENTIONED DIFFERENT LENDING 1131 00:48:09,720 --> 00:48:10,120 SITES. 1132 00:48:10,120 --> 00:48:11,400 DIFFERENT COLLABORATIONS 1133 00:48:11,400 --> 00:48:13,200 DIFFERENT SITES ON THE MOON, 1134 00:48:13,200 --> 00:48:16,120 ALSO SHOW SLIGHT DIFFERENCES IN 1135 00:48:16,120 --> 00:48:18,800 RATIO OF IRON OXIDE, MAGNESIUM O 1136 00:48:18,800 --> 00:48:20,080 SIDE, CALCIUM OXIDE I MENTIONED 1137 00:48:20,080 --> 00:48:20,600 EARLIER. 1138 00:48:20,600 --> 00:48:23,080 SO I HOPE THAT ANSWERS YOUR 1139 00:48:23,080 --> 00:48:23,360 QUESTION. 1140 00:48:23,360 --> 00:48:24,160 HOPE IT IS MAKING SENSE. 1141 00:48:24,160 --> 00:48:26,400 THANK YOU. 1142 00:48:26,400 --> 00:48:30,600 >> IS TH THANK YOU. 1143 00:48:30,600 --> 00:48:34,400 QUESTION FOR DR. LEE. 1144 00:48:34,400 --> 00:48:35,960 WHAT IS THE HUMAN CELL LINE DOES 1145 00:48:35,960 --> 00:48:39,920 IT EXPRESS PARP 2 IN NUCLEUS AND 1146 00:48:39,920 --> 00:48:40,360 MITOCHONDRIA? 1147 00:48:40,360 --> 00:48:42,520 >> YEAH. 1148 00:48:42,520 --> 00:48:44,320 THAT IS A VERY GOOD QUESTION. 1149 00:48:44,320 --> 00:48:46,960 WE USE HELO CELLS AND WE USE 1150 00:48:46,960 --> 00:48:51,280 CONTROL AS HELO WILD TYPE AND WE 1151 00:48:51,280 --> 00:48:53,640 USE HELO KNOCK OUT CELLS. 1152 00:48:53,640 --> 00:48:56,440 MOST EXPERIMENTAL RESULTS ARE 1153 00:48:56,440 --> 00:48:59,680 ACROSS CONFIRMED IN CELLS 1154 00:48:59,680 --> 00:49:04,520 KNOCKED DOWN WITH PARP1. 1155 00:49:04,520 --> 00:49:07,600 SO WE DO NOT KNOW IF IT 1156 00:49:07,600 --> 00:49:11,640 EXPRESSES PARP 2 IN NUCLEUS OR 1157 00:49:11,640 --> 00:49:14,440 MITOCHONDRIA, BUT I CAN SAY 1158 00:49:14,440 --> 00:49:19,280 THERE IS A LITTLE BIT OF IMPACT 1159 00:49:19,280 --> 00:49:20,600 OF TREATMENT THOUGH THERE WAS NO 1160 00:49:20,600 --> 00:49:21,080 PARP1. 1161 00:49:21,080 --> 00:49:27,240 SO PARP1 KNOCK OUT CELLS ARE 1162 00:49:27,240 --> 00:49:28,280 RESPONDING TO (INAUDIBLE), WHICH 1163 00:49:28,280 --> 00:49:29,280 MEANS THERE COULD BE PARP 2 OR 1164 00:49:29,280 --> 00:49:31,920 3. 1165 00:49:31,920 --> 00:49:34,120 >> THANK YOU, DR. YANG DO YOU 1166 00:49:34,120 --> 00:49:35,600 WANT TO READ THE NEXT ONE? 1167 00:49:35,600 --> 00:49:38,040 >> I WILL GO WITH THE NEXT ONE. 1168 00:49:38,040 --> 00:49:40,000 THE QUESTION IS FROM MATTHEW 1169 00:49:40,000 --> 00:49:41,720 LONGLY. 1170 00:49:41,720 --> 00:49:44,040 THIS IS FOR MATT. 1171 00:49:44,040 --> 00:49:44,760 NICE TALK. 1172 00:49:44,760 --> 00:49:47,240 CAN YOUR EXPERIMENTAL SYSTEM BE 1173 00:49:47,240 --> 00:49:49,080 ADAPTED TO ARTIFICIAL 1174 00:49:49,080 --> 00:49:51,200 REPLICATION OR OTHER SPECIALIZED 1175 00:49:51,200 --> 00:49:53,880 STRUCTURE CONTAIN SINGLE 1176 00:49:53,880 --> 00:49:54,480 STRANDED DNA? 1177 00:49:54,480 --> 00:49:57,600 >> THANKS FOR THE QUESTION. 1178 00:49:57,600 --> 00:49:58,680 YOU ARE HITTING ON SOMETHING 1179 00:49:58,680 --> 00:50:01,840 THAT WAS A BIG CONCERN OF OURS 1180 00:50:01,840 --> 00:50:05,200 WHEN YOU HAVE NUCLEAR EXTRACT 1181 00:50:05,200 --> 00:50:07,160 EXPOSED TO DNA SUBSTRATE SO 1182 00:50:07,160 --> 00:50:09,480 TIGHT ROPE ANY KNEE COLLIEIUS 1183 00:50:09,480 --> 00:50:12,440 CUT THE DNA WILL BREAK AND WE 1184 00:50:12,440 --> 00:50:14,720 WON'T BE ABLE TO -- ANY MORE. 1185 00:50:14,720 --> 00:50:16,240 AT FIRST DOUBLE STRAND DNA WE 1186 00:50:16,240 --> 00:50:19,680 WERE SUSPICIOUS HOW LONG CAN WE 1187 00:50:19,680 --> 00:50:21,600 IMAGE, WE CAN GET 15 MINUTES OR 1188 00:50:21,600 --> 00:50:22,760 LONGER WITH DOUBLE STRAND DNA. 1189 00:50:22,760 --> 00:50:25,120 WE ALSO TRIED TO EXTRACT SINGLE 1190 00:50:25,120 --> 00:50:26,480 STRAND DNA AND TO CONFIRM THERE 1191 00:50:26,480 --> 00:50:28,280 IS ONLY A SINGLE CUT YOU LOSE 1192 00:50:28,280 --> 00:50:29,480 YOUR IMAGE. 1193 00:50:29,480 --> 00:50:31,880 EVERYONE ON SINGLE STRAND DNA IT 1194 00:50:31,880 --> 00:50:33,640 DOES SEEM WE CAN IMAGE FOR FIVE 1195 00:50:33,640 --> 00:50:35,480 OR TEN MINUTES AT LEAST. 1196 00:50:35,480 --> 00:50:37,360 SO I WOULD SAY YES WE CAN ADAPT 1197 00:50:37,360 --> 00:50:39,280 THAT TO THE ARTIFICIAL 1198 00:50:39,280 --> 00:50:40,600 REPLICATION. 1199 00:50:40,600 --> 00:50:41,920 BUT IT IS A LITTLE MORE 1200 00:50:41,920 --> 00:50:43,840 CHALLENGING BECAUSE YOU HAVE TO 1201 00:50:43,840 --> 00:50:45,200 WITH REPLICATION YOU HAVE THREE 1202 00:50:45,200 --> 00:50:48,320 ENDS SO YOU NEED THREE BEADS YOU 1203 00:50:48,320 --> 00:50:50,720 NEED SOME WAY TO GENERATE THAT 1204 00:50:50,720 --> 00:50:51,560 SUBSTRATE WHICH WOULD BE 1205 00:50:51,560 --> 00:50:52,560 SOMEWHAT COMPLICATED. 1206 00:50:52,560 --> 00:50:57,400 IT IS A GREAT DIRECTION TO GO. 1207 00:50:57,400 --> 00:51:00,840 >> DR. DIMPLE, YOU ARE A HOST 1208 00:51:00,840 --> 00:51:02,520 NOW, CAN YOU UNMUTE AND ASK YOUR 1209 00:51:02,520 --> 00:51:02,840 QUESTION? 1210 00:51:02,840 --> 00:51:05,120 >> SURE. 1211 00:51:05,120 --> 00:51:07,440 I HAVE QUESTION FOR TWO OF YOU. 1212 00:51:07,440 --> 00:51:10,080 FIRST ABOUT THE CORRELATION, 1213 00:51:10,080 --> 00:51:13,840 MITOCHONDRIAL PARYLATION IS THAT 1214 00:51:13,840 --> 00:51:14,960 AFFECTED BY TREATING CELLS WITH 1215 00:51:14,960 --> 00:51:16,480 DNA DAMAGING AGENTS? 1216 00:51:16,480 --> 00:51:19,040 >> OUR MAIN PURPOSE OF THIS 1217 00:51:19,040 --> 00:51:21,160 RESEARCH IS TO IDENTIFY 1218 00:51:21,160 --> 00:51:24,280 MITOCHONDRIAL PARP1 EXISTENCE. 1219 00:51:24,280 --> 00:51:27,240 THAT IS WHY USE MOST PURIFIED 1220 00:51:27,240 --> 00:51:28,120 MITE CHON TRIA FOR MOST 1221 00:51:28,120 --> 00:51:31,280 EXPERIMENTAL SETTING. 1222 00:51:31,280 --> 00:51:33,520 UNFORTUNATELY I DIDN'T PERFORM 1223 00:51:33,520 --> 00:51:35,880 INTACT CELLS AND (INAUDIBLE) DNA 1224 00:51:35,880 --> 00:51:39,480 DAMAGE AGENTS BUT AT LEAST WHEN 1225 00:51:39,480 --> 00:51:41,760 WE TRY TO TREAT ISOLATED 1226 00:51:41,760 --> 00:51:45,920 MITOCHONDRIA WITH HYDROGEN 1227 00:51:45,920 --> 00:51:48,120 PEROXIDE IT DIDN'T REACT. 1228 00:51:48,120 --> 00:51:52,000 >> THANK YOU. 1229 00:51:52,000 --> 00:51:56,040 QUESTION FOR THAT, IS 1230 00:51:56,040 --> 00:51:57,440 UBIQUITINATION ACTIVE UNDER YOUR 1231 00:51:57,440 --> 00:51:57,960 CONDITIONS? 1232 00:51:57,960 --> 00:51:59,440 IF IT IS NOT CAN YOU MAKE IT 1233 00:51:59,440 --> 00:52:02,480 ACTIVE AND SEE WHAT HAPPENS? 1234 00:52:02,480 --> 00:52:04,280 >> THANKS, REALLY GREAT 1235 00:52:04,280 --> 00:52:07,120 QUESTION. 1236 00:52:07,120 --> 00:52:08,680 WE ARE LOOKING AT MAYBE GETTING 1237 00:52:08,680 --> 00:52:12,520 A FLUORESCENTLY TAGGED GFP 1238 00:52:12,520 --> 00:52:14,000 UBIQUITIN CONSTRUCT, WE DON'T 1239 00:52:14,000 --> 00:52:15,080 KNOW WHETHER OR NOT IT IS 1240 00:52:15,080 --> 00:52:16,560 UBIQUITINATED OR NOT. 1241 00:52:16,560 --> 00:52:18,760 SO IT IS IN THE NUCLEAR EXTRACT. 1242 00:52:18,760 --> 00:52:21,080 IN PRINCIPLE COULD BE ACTIVELY 1243 00:52:21,080 --> 00:52:21,920 UBIQUITINATED. 1244 00:52:21,920 --> 00:52:24,520 MAYBE EXPLAIN DIFFERENT 1245 00:52:24,520 --> 00:52:25,120 LIFETIMES. 1246 00:52:25,120 --> 00:52:29,320 SO WE HAVE SUCH A LONG DISPARATE 1247 00:52:29,320 --> 00:52:30,080 POPULATION BETWEEN HUNDRED 1248 00:52:30,080 --> 00:52:31,640 SECONDS AND .1 SECONDS SO MAYBE 1249 00:52:31,640 --> 00:52:35,440 ONE OF THOSE POPULATIONS IS A 1250 00:52:35,440 --> 00:52:35,960 UBIQUITINATED. 1251 00:52:35,960 --> 00:52:38,680 WE ARE INTERESTED IN THE SAME, 1252 00:52:38,680 --> 00:52:39,880 SO THANK YOU. 1253 00:52:39,880 --> 00:52:42,320 -- INTERESTED IN PURSUING THAT. 1254 00:52:42,320 --> 00:52:43,000 THANK YOU. 1255 00:52:43,000 --> 00:52:44,040 >> WHY DON'T YOU GO. 1256 00:52:44,040 --> 00:52:45,720 >>LY GO TO NEXT ONE. 1257 00:52:45,720 --> 00:52:49,520 THIS QUESTION IS FROM JOE JONG. 1258 00:52:49,520 --> 00:52:52,640 THIS IS TO MATT TOO, GREAT TALK. 1259 00:52:52,640 --> 00:52:56,200 DO YOU SEE THE BACKGROUND LEVEL 1260 00:52:56,200 --> 00:52:57,480 UVDDB PLAY A ROLE IN YOUR 1261 00:52:57,480 --> 00:52:59,800 RESULTS HAVING CONSIDERED THAT 1262 00:52:59,800 --> 00:53:02,400 YOU KNOCK IN FLUORESCENT TAG? 1263 00:53:02,400 --> 00:53:03,480 >> THANKS. 1264 00:53:03,480 --> 00:53:05,520 THAT IS A REALLY IMPORTANT 1265 00:53:05,520 --> 00:53:08,640 CONCEPT WE ARE FINDING IS THAT 1266 00:53:08,640 --> 00:53:11,280 PROTEINS UNDERGO WHAT IS CALLED 1267 00:53:11,280 --> 00:53:12,880 FACILITATED ASSOCIATION SO IF 1268 00:53:12,880 --> 00:53:14,840 YOU HAVE A HIGH POPULATION OF 1269 00:53:14,840 --> 00:53:16,520 UNLABELED PROTEIN COMPETING WITH 1270 00:53:16,520 --> 00:53:19,960 FLUORESCENT LABEL IT SHORTENS 1271 00:53:19,960 --> 00:53:21,400 THE BINDING LIFETIME AND WE DID 1272 00:53:21,400 --> 00:53:22,960 EXPERIMENT IF YOU INCLUDE 1273 00:53:22,960 --> 00:53:26,800 PURIFIED UVDDB WITH THE 1274 00:53:26,800 --> 00:53:28,560 EXPERIMENT IT DOES SHORTEN THE 1275 00:53:28,560 --> 00:53:29,200 LIFETIME. 1276 00:53:29,200 --> 00:53:32,120 WE ARE INTERESTED IN DOING A 1277 00:53:32,120 --> 00:53:33,000 KNOCK IN P FLUORESCENCE TAG. 1278 00:53:33,000 --> 00:53:36,360 WE ALSO HAVE PRELIMINARY DATA 1279 00:53:36,360 --> 00:53:38,040 WHERE WE HAVE DONE A STABLE CELL 1280 00:53:38,040 --> 00:53:41,440 LINE EXPRESSING DDB 2 TAG NEAR 1281 00:53:41,440 --> 00:53:43,440 ENDOGENOUS LEVELS SO NOT 1282 00:53:43,440 --> 00:53:45,120 OVEREXPRESSED AND IT DOES BEHAVE 1283 00:53:45,120 --> 00:53:48,560 DIFFERENTLY TO THE OVEREXPRESSED 1284 00:53:48,560 --> 00:53:49,920 VERSION BECAUSE IT HAS THESE 1285 00:53:49,920 --> 00:53:52,520 OTHER PARTNERS THAT ARE CHANGING 1286 00:53:52,520 --> 00:53:53,120 THE CHARACTER. 1287 00:53:53,120 --> 00:53:58,160 IT WILL BE FUN TO UNRAVEL THOSE 1288 00:53:58,160 --> 00:53:59,280 COMPLEXITY. 1289 00:53:59,280 --> 00:54:02,120 >> DR. LEE, HAVE YOU SHARED 1290 00:54:02,120 --> 00:54:03,880 RESULTS WITH NORMAL CELLS WITH 1291 00:54:03,880 --> 00:54:06,320 HELO CELLS, HAVE YOU TESTED ANY 1292 00:54:06,320 --> 00:54:10,840 CELLS WITH DISEASES MITOCHONDRIA 1293 00:54:10,840 --> 00:54:14,840 LIKE (INAUDIBLE) SYNDROME FOR 1294 00:54:14,840 --> 00:54:15,120 INSTANCE? 1295 00:54:15,120 --> 00:54:17,520 >> THAT WAS A INITIAL PLAN. 1296 00:54:17,520 --> 00:54:24,040 SO I TESTED ISOLATED 1297 00:54:24,040 --> 00:54:25,680 MITOCHONDRIA FROM COCAINE 1298 00:54:25,680 --> 00:54:27,120 SYNDROME PATIENTS AND THEY SEEM 1299 00:54:27,120 --> 00:54:31,520 TO BE LATE ELEVATED BUT I DIDN'T 1300 00:54:31,520 --> 00:54:39,360 INCLUDE THOSE IN THE DATA. 1301 00:54:39,360 --> 00:54:41,280 >> THANK YOU. 1302 00:54:41,280 --> 00:54:42,520 JAMIE, I HAVE A QUESTION FOR 1303 00:54:42,520 --> 00:54:43,960 YOU. 1304 00:54:43,960 --> 00:54:46,440 THIS IS A DNA REPAIR INTEREST 1305 00:54:46,440 --> 00:54:48,440 GROUP, YOU ARE SHOWING A LOT OF 1306 00:54:48,440 --> 00:54:49,600 INFORMATION ABOUT DAMAGE, DO YOU 1307 00:54:49,600 --> 00:54:51,680 HAVE ANYTHING ABOUT REPAIR OF 1308 00:54:51,680 --> 00:54:54,680 THAT DAMAGE? 1309 00:54:54,680 --> 00:54:56,080 >> YEAH. 1310 00:54:56,080 --> 00:54:57,520 I HAVEN'T GO INTO THAT BECAUSE I 1311 00:54:57,520 --> 00:55:02,120 HAVEN'T DONE A LOT OF REPAIR BUT 1312 00:55:02,120 --> 00:55:04,000 I HAVE BEEN EXPECTING, I'M GOING 1313 00:55:04,000 --> 00:55:07,760 TO PURSUE TESTING AND IN FACT 1314 00:55:07,760 --> 00:55:10,600 BER COULD BE THE MAIN MECHANISM 1315 00:55:10,600 --> 00:55:16,400 OF DNA REPAIR BECAUSE AS I HAVE 1316 00:55:16,400 --> 00:55:17,920 SHOWN -- MAIN MECHANISM OF 1317 00:55:17,920 --> 00:55:22,760 TOXICITY OF LUNAR SIMULANTS. 1318 00:55:22,760 --> 00:55:25,400 CR IS ALSO ASSOCIATED WITH KIND 1319 00:55:25,400 --> 00:55:29,960 OF OXIDATIVE DAMAGE CAUSED BY 1320 00:55:29,960 --> 00:55:31,520 FACTORS IN THE ENVIRONMENT 1321 00:55:31,520 --> 00:55:32,040 ITSELF. 1322 00:55:32,040 --> 00:55:33,920 SO THAT IS WHAT I'M THINKING BUT 1323 00:55:33,920 --> 00:55:35,880 YEAH, I HAVEN'T -- I MEAN 1324 00:55:35,880 --> 00:55:38,840 BECAUSE LIMITED TIME I DID NOT 1325 00:55:38,840 --> 00:55:39,400 BRING THAT YET. 1326 00:55:39,400 --> 00:55:40,960 SO THANKS FOR THE QUESTION. 1327 00:55:40,960 --> 00:55:42,080 >> THANK YOU. 1328 00:55:42,080 --> 00:55:43,680 WE DON'T HAVE ANY MORE QUESTIONS 1329 00:55:43,680 --> 00:55:46,040 IN THE CHAT. 1330 00:55:46,040 --> 00:55:47,440 LET ME THANK THE SPEAKERS FOR 1331 00:55:47,440 --> 00:55:48,600 THE WONDERFUL PRESENTATIONS 1332 00:55:48,600 --> 00:55:49,720 TODAY. 1333 00:55:49,720 --> 00:55:52,840 AND REMIND PEOPLE IN THE 1334 00:55:52,840 --> 00:55:54,920 AUDIENCE THAT YOU CAN JOIN THE 1335 00:55:54,920 --> 00:55:57,280 LIST SERVE FOR THE DNA REPAIR 1336 00:55:57,280 --> 00:55:58,840 INTEREST GROUP AND FIND OUT 1337 00:55:58,840 --> 00:56:00,680 ABOUT THE VIDEO CONFERENCES, 1338 00:56:00,680 --> 00:56:03,080 OTHER ONES, ABOUT MEETINGS, JOBS 1339 00:56:03,080 --> 00:56:04,720 AND MUCH MORE. 1340 00:56:04,720 --> 00:56:09,040 THE ANNOUNCEMENT YOU GOT YOU CAN 1341 00:56:09,040 --> 00:56:13,720 SEND ME AN EMAIL AT KRAMERK@NIH 1342 00:56:13,720 --> 00:56:16,040 TO BE INCLUDED IN THE LIST 1343 00:56:16,040 --> 00:56:16,640 THERE. 1344 00:56:16,640 --> 00:56:18,400 OR THERE WAS A LINK TO THE DNA 1345 00:56:18,400 --> 00:56:19,800 REPAIR INTEREST GROUP WEBSITE IN 1346 00:56:19,800 --> 00:56:21,120 THE EMAIL THAT YOU RECEIVED 1347 00:56:21,120 --> 00:56:23,400 TODAY. 1348 00:56:23,400 --> 00:56:27,040 SO THANK YOU SO MUCH. 1349 00:56:27,040 --> 00:56:27,840 GOOD LUCK. 1350 00:56:27,840 --> 00:00:00,000 THANK YOU.