1 00:00:05,560 --> 00:00:10,560 IT IS A TREMENDOUS PLEASURE TO 2 00:00:10,560 --> 00:00:15,960 ENTINTRODUCE YOSHI KAWAOKA 3 00:00:15,960 --> 00:00:20,680 WITHOUT KNOWING HIM PERSONALLY 4 00:00:20,680 --> 00:00:25,320 WHEN I DID TRAINING IN THE LAB 5 00:00:25,320 --> 00:00:30,200 IN GERMANY AND HE WAS INFECTIOUS 6 00:00:30,200 --> 00:00:32,560 GROUP. 7 00:00:32,560 --> 00:00:34,440 GOT TO KNOW HIM AS COLLABORATOR 8 00:00:34,440 --> 00:00:36,200 WHEN WE STARTED TO WORK ON 1918 9 00:00:36,200 --> 00:00:44,240 FLU AND EXB EBOLA AND WE BECAME 10 00:00:44,240 --> 00:00:48,640 FRIENDS OVER THAT COLLABORATION 11 00:00:48,640 --> 00:00:49,640 UNTIL TODAY. 12 00:00:49,640 --> 00:00:54,960 DR. YOSHI HAS A DVM FROM 13 00:00:54,960 --> 00:00:56,760 UNIVERSITY IN JAPAN, I LEAVE THE 14 00:00:56,760 --> 00:00:57,360 YEARS OUT. 15 00:00:57,360 --> 00:01:02,760 WE ARE ALL VERY YOUNG STILL. 16 00:01:02,760 --> 00:01:04,840 SO HIS RESEARCH CAREER IS A 17 00:01:04,840 --> 00:01:05,760 TREMENDOUS JOURNEY. 18 00:01:05,760 --> 00:01:11,760 AS I SAID HE DID A POST DOC IN 19 00:01:11,760 --> 00:01:13,560 MEMPHIS AND WENT TO ASSISTANT 20 00:01:13,560 --> 00:01:16,280 MEMBER ASSOCIATE MEMBER 21 00:01:16,280 --> 00:01:17,120 ASSOCIATE PROFESSORSHIP POSITION 22 00:01:17,120 --> 00:01:18,520 AND BECAME FULL PROFESSOR 23 00:01:18,520 --> 00:01:21,800 UNIVERSITY OF MADISON WISCONSIN, 24 00:01:21,800 --> 00:01:25,040 WISCONSIN MADISON WHERE HE IS 25 00:01:25,040 --> 00:01:29,400 STILL TODAY AND BECAME A UNIQUE 26 00:01:29,400 --> 00:01:32,600 SITUATION FOR ME AT LEAST AND I 27 00:01:32,600 --> 00:01:35,800 ALWAYS ADMIRE THIS THROUGHOUT 28 00:01:35,800 --> 00:01:36,840 FULL PROFESSOR UNIVERSITY OF 29 00:01:36,840 --> 00:01:37,560 TOKYO. 30 00:01:37,560 --> 00:01:40,600 ONLY TWO YEARS AFTER HE JOINED 31 00:01:40,600 --> 00:01:42,320 MEDICINE. 32 00:01:42,320 --> 00:01:44,200 HE KEPT THOSE TWO AFFILIATIONS 33 00:01:44,200 --> 00:01:47,520 THROUGHOUT HIS CAREER GOING BACK 34 00:01:47,520 --> 00:01:49,840 AND FORTH BETWEEN THOSE TWO 35 00:01:49,840 --> 00:01:51,040 CONTINENTS AND NEVER GETTING 36 00:01:51,040 --> 00:01:55,480 TIRED OF FLYING BACK AND FORTH 37 00:01:55,480 --> 00:01:57,080 THOUGH HE TOLD ME TODAY HE IS A 38 00:01:57,080 --> 00:01:58,960 LITTLE TIRED OF IT AND THAT OF 39 00:01:58,960 --> 00:02:02,400 COURSE WAS HAMPERED QUITE A BET 40 00:02:02,400 --> 00:02:03,960 BY SARS COV-2 EMERGENCE AND 41 00:02:03,960 --> 00:02:07,360 COVID PANDEMIC. 42 00:02:07,360 --> 00:02:09,440 JUST RESENTLY HE BECAME DIRECTOR 43 00:02:09,440 --> 00:02:11,120 OF RESEARCH CENTER FOR GLOBAL 44 00:02:11,120 --> 00:02:12,880 VIRAL INFECTIONS NATIONAL CENTER 45 00:02:12,880 --> 00:02:15,520 FOR GLOBAL HEALTH AND MEDICINE 46 00:02:15,520 --> 00:02:19,640 IN JAPAN AND THIS IS HIS LATEST 47 00:02:19,640 --> 00:02:21,240 ENDEAVOR, HE HAD MANY MORE 48 00:02:21,240 --> 00:02:22,720 AFFILIATIONS OVER THE YEARS AND 49 00:02:22,720 --> 00:02:23,640 STILL MAY HAVE MORE THAT I DON'T 50 00:02:23,640 --> 00:02:26,320 KNOW ABOUT. 51 00:02:26,320 --> 00:02:27,880 IN TERMS OF SCIENTIFIC 52 00:02:27,880 --> 00:02:30,520 PRODUCTIVITY, IT IS ABSOLUTELY 53 00:02:30,520 --> 00:02:35,360 FASCINATING, PUBMED LISTS 700 54 00:02:35,360 --> 00:02:37,520 PEER REVIEWED PUBLICATIONS PLUS 55 00:02:37,520 --> 00:02:42,160 150 MORE CONTRIBUTIONS, MANY 56 00:02:42,160 --> 00:02:43,760 PATENTS, UNCOUNTABLE NATIONAL 57 00:02:43,760 --> 00:02:44,840 AND INTERNATIONAL INVITED 58 00:02:44,840 --> 00:02:45,600 PRESENTATIONS. 59 00:02:45,600 --> 00:02:53,160 AND MANY, MANY AWARDS TO MENTION 60 00:02:53,160 --> 00:02:57,200 A FEW, THE ROBERT COUGH AWARDED 61 00:02:57,200 --> 00:03:01,160 THE PRESTIGIOUS GERMAN AWARDS. 62 00:03:01,160 --> 00:03:03,640 THE MEDAL OF HONOR, PURPLE 63 00:03:03,640 --> 00:03:06,360 RIBBON FROM JAPAN AND AWARD FROM 64 00:03:06,360 --> 00:03:10,480 JAPAN ACADEMY AWARD FROM THE 65 00:03:10,480 --> 00:03:11,520 EMPEROR OF JAPAN. 66 00:03:11,520 --> 00:03:13,280 BECAME MEMBER OF THE NATIONAL 67 00:03:13,280 --> 00:03:18,280 ACADEMY SCIENCES IN THE U.S. IN 68 00:03:18,280 --> 00:03:21,560 2013 AND IN 2015, THE COLLEGE 69 00:03:21,560 --> 00:03:22,760 FINALLY PRIZE FOR MICROBIOLOGY. 70 00:03:22,760 --> 00:03:30,320 AS I SAID THIS IS A FEW OF HIS 71 00:03:30,320 --> 00:03:31,360 HONORS AND AWARDS. 72 00:03:31,360 --> 00:03:35,000 HIS PROFESSIONAL INTEREST IS 73 00:03:35,000 --> 00:03:36,880 INFLUENZA. 74 00:03:36,880 --> 00:03:38,760 HE DEVELOPED REVERSE GENETICS TO 75 00:03:38,760 --> 00:03:41,120 STUDY PATHOGENESIS DEVELOP 76 00:03:41,120 --> 00:03:43,320 VACCINES AND MANY OTHER ASPECTS 77 00:03:43,320 --> 00:03:45,440 OF HIS AREAS OF RESEARCH AND 78 00:03:45,440 --> 00:03:47,960 STILL DOES THAT TODAY. 79 00:03:47,960 --> 00:03:51,120 HE IS ALSO DONE TREMENDOUS WORK 80 00:03:51,120 --> 00:03:55,800 ON EBOLA STARTING WITH REVERSE 81 00:03:55,800 --> 00:03:56,800 GENETICS CREATING A VACCINE THAT 82 00:03:56,800 --> 00:03:59,200 IS IN CLINICAL TRIALS, STUDYING 83 00:03:59,200 --> 00:04:00,800 PATHOGENESIS, AND EVEN DOING 84 00:04:00,800 --> 00:04:02,560 FIELD STUDIES WHICH HE STILL 85 00:04:02,560 --> 00:04:08,480 CONTINUES TO DO IN WEST AFRICA. 86 00:04:08,480 --> 00:04:09,480 SOMETHING I BELIEVE VERY NEW TO 87 00:04:09,480 --> 00:04:15,200 HIM AT THE TIME IT STARTED. 88 00:04:15,200 --> 00:04:18,360 OF COURSE AS MANY OF INVOLVED IN 89 00:04:18,360 --> 00:04:20,640 SARS COV-2 AND WE WILL HEAR THAT 90 00:04:20,640 --> 00:04:23,680 TODAY IN HIS PRESENTATION. 91 00:04:23,680 --> 00:04:26,600 IT IS AN HONOR AND PLEASURE TO 92 00:04:26,600 --> 00:04:28,920 BE HERE AND INTRODUCE YASHI. 93 00:04:28,920 --> 00:04:30,320 DON'T WANT TO STEAL MORE TIME 94 00:04:30,320 --> 00:04:31,160 FROM YOUR TALK. 95 00:04:31,160 --> 00:04:34,760 WE ARE LOOKING FORWARD TO 96 00:04:34,760 --> 00:04:37,840 FASCINATING JOURNEY THROUGH SARS 97 00:04:37,840 --> 00:04:38,880 COV-2, GO AHEAD. 98 00:04:38,880 --> 00:04:41,680 THANKS FOR COMING AND ACCEPTING 99 00:04:41,680 --> 00:04:43,920 THE INVITATION TO SPEAK. 100 00:04:43,920 --> 00:04:45,480 >>THANK YOU FOR THAT NICE 101 00:04:45,480 --> 00:04:45,840 INTRODUCTION. 102 00:04:45,840 --> 00:04:51,400 I WOULD LIKE TO ALSO THANK FOR 103 00:04:51,400 --> 00:04:52,480 INVITING ME TO TALK TO YOU SOME 104 00:04:52,480 --> 00:04:58,880 OF THE WORK. 105 00:04:58,880 --> 00:04:59,960 TODAY AS HINES MENTIONED, I GET 106 00:04:59,960 --> 00:05:02,560 TO TALK ABOUT SARS COV-2 WHAT WE 107 00:05:02,560 --> 00:05:03,600 HAVE BEEN WORKING ON SO I'M 108 00:05:03,600 --> 00:05:10,000 GOING TO COVER FOUR TOPICS. 109 00:05:10,000 --> 00:05:10,720 FIRST ANIMAL MODELS. 110 00:05:10,720 --> 00:05:12,280 AT THE BEGINNING OF THIS 111 00:05:12,280 --> 00:05:15,840 PANDEMIC WE NEEDED TO ANIMAL 112 00:05:15,840 --> 00:05:17,800 MODELS TO STUDY. 113 00:05:17,800 --> 00:05:26,280 SARS COV-2 WE INFECTED DIFFERENT 114 00:05:26,280 --> 00:05:28,320 ANIMAL SPECIES AND FOUND THE 115 00:05:28,320 --> 00:05:31,280 PROTOTYPE SARS COV-2 DIDN'T 116 00:05:31,280 --> 00:05:33,080 REPLICATE WELL IN MARMOSET AND 117 00:05:33,080 --> 00:05:36,520 REGULAR MICE. 118 00:05:36,520 --> 00:05:42,000 AS YOU KNOW SOME OF THE RECENT 119 00:05:42,000 --> 00:05:44,160 SOME OF THE VARIANT VIRUSES CAN 120 00:05:44,160 --> 00:05:47,200 INFECT REGULAR MICE BUT WE ALSO 121 00:05:47,200 --> 00:05:50,040 FOUND THAT THE HOSTS ARE HIGHLY 122 00:05:50,040 --> 00:05:52,560 SUSCEPTIBLE. 123 00:05:52,560 --> 00:05:57,520 IN FACT, CATS WHO WENT WITH THE 124 00:05:57,520 --> 00:05:58,880 COVID-19 PATIENT BECAME PCR 125 00:05:58,880 --> 00:06:03,560 POSITIVE. 126 00:06:03,560 --> 00:06:08,760 SO WE INOCULATED CATS WITH SARS 127 00:06:08,760 --> 00:06:11,040 COV-2 THEN EXAMINED WHETHER THE 128 00:06:11,040 --> 00:06:14,240 VIRUS REPLICATES IN CATS OR NOT 129 00:06:14,240 --> 00:06:17,280 AND THEN CO-HOUSED UNINFECTED 130 00:06:17,280 --> 00:06:19,120 CAT TO SEE WHETHER THE DEVICE 131 00:06:19,120 --> 00:06:21,600 CAN BE TRANSMITTED FROM 132 00:06:21,600 --> 00:06:23,040 INOCULATED CAT TO UNINFECTED 133 00:06:23,040 --> 00:06:27,440 CAT. 134 00:06:27,440 --> 00:06:29,160 WE FOUND THAT THE VIRUS 135 00:06:29,160 --> 00:06:32,160 REPLICATES QUITE WELL IN THE 136 00:06:32,160 --> 00:06:33,360 INOCULATED CAT AND THE VIRUS CAN 137 00:06:33,360 --> 00:06:36,120 BE TRANSMITTED TO CO-HOUSED 138 00:06:36,120 --> 00:06:38,680 UNINFECTED CAT WITH TWO TO FIVE 139 00:06:38,680 --> 00:06:43,760 DAYS OF CO-HOUSING. 140 00:06:43,760 --> 00:06:45,640 WE FOUND THE VIRUS REPLICATES 141 00:06:45,640 --> 00:06:49,480 WELL IN MORE THAN 10,000 142 00:06:49,480 --> 00:06:50,720 INFECTIOUS UNITS IN THE NASAL 143 00:06:50,720 --> 00:06:53,200 SWABS AND WHAT IS INTERESTING IS 144 00:06:53,200 --> 00:06:55,200 THAT NONE OF THESE ANIMALS 145 00:06:55,200 --> 00:06:59,720 SHOWED NO SYMPTOMS. 146 00:06:59,720 --> 00:07:01,880 SO WHAT THIS TELLS US IS THAT 147 00:07:01,880 --> 00:07:06,560 THE VIRUS CAN BE TRANSMITTED 148 00:07:06,560 --> 00:07:09,000 FROM HUMANS TO CAT AND TRANSMIT 149 00:07:09,000 --> 00:07:10,800 AMONG CATS WITHOUT CAUSING ANY 150 00:07:10,800 --> 00:07:12,160 SYMPTOMS. 151 00:07:12,160 --> 00:07:13,880 SO WE DON'T KNOW WHETHER THE 152 00:07:13,880 --> 00:07:16,000 VIRUS CAN BE TRANSMITTED FROM 153 00:07:16,000 --> 00:07:20,000 CATS TO HUMANS. 154 00:07:20,000 --> 00:07:24,360 NOW, WE LET SOME OF THESE CATS 155 00:07:24,360 --> 00:07:27,720 FOR 28 DAYS THEN EUTHANIZED AND 156 00:07:27,720 --> 00:07:28,720 EXAMINED LUNG PATHOLOGY. 157 00:07:28,720 --> 00:07:31,480 SO THIS WORK WAS DONE BY 158 00:07:31,480 --> 00:07:32,400 (INDISCERNIBLE) IN OUR 159 00:07:32,400 --> 00:07:32,840 DEPARTMENT. 160 00:07:32,840 --> 00:07:35,680 AND SHE FOUND THAT THE 161 00:07:35,680 --> 00:07:37,480 PATHOLOGICAL CHANGES LASTED FOR 162 00:07:37,480 --> 00:07:40,680 28 DAYS IN THE LUNGS. 163 00:07:40,680 --> 00:07:42,120 SO THIS MAY ALSO HAPPEN IN 164 00:07:42,120 --> 00:07:49,960 HUMANS. 165 00:07:49,960 --> 00:07:56,720 SO NOW WE INFECTED HAMSTERS WITH 166 00:07:56,720 --> 00:07:59,640 SARS COV-2 THEN EXAMINED THE 167 00:07:59,640 --> 00:08:02,640 VIRUS DATA ON DAY 3 AND FOUND 168 00:08:02,640 --> 00:08:05,520 THAT THE MORE THAN 10 TO THE 8TH 169 00:08:05,520 --> 00:08:07,680 INFECTIOUS UNITS IN NASAL 170 00:08:07,680 --> 00:08:09,360 TERMINATE IN THE LUNGS. 171 00:08:09,360 --> 00:08:14,440 ON THE 6 VIRUS TITER DROPPED BY 172 00:08:14,440 --> 00:08:16,360 TWO LOG UNITS. 173 00:08:16,360 --> 00:08:18,360 AND WE DIDN'T FIND ANY VIRUS IN 174 00:08:18,360 --> 00:08:23,800 KIDNEY, INTESTINE AND BLOOD. 175 00:08:23,800 --> 00:08:28,600 WE ALSO EXAMINED LUNG LESIONS BY 176 00:08:28,600 --> 00:08:32,440 USING MICROCD. 177 00:08:32,440 --> 00:08:34,440 BEFORE INFECTION AND SEVEN DAYS 178 00:08:34,440 --> 00:08:39,480 POST INFECTION AND 16 DAYS POST 179 00:08:39,480 --> 00:08:41,240 INFECTION OF THE SAME ANIMAL. 180 00:08:41,240 --> 00:08:43,520 AS YOU CAN SEE LUNG LESIONS ARE 181 00:08:43,520 --> 00:08:49,960 MOST PROMINENT ON DAY 7 MOSTLY 182 00:08:49,960 --> 00:08:53,040 BECAUSE OF BASIC STAIN. 183 00:08:53,040 --> 00:08:56,800 SOME IN OUR DEPARTMENT EXAMINED 184 00:08:56,800 --> 00:08:58,800 THIS CT SCAN AND FOUND THAT 185 00:08:58,800 --> 00:09:00,640 HAMSTERS INFECTED WITH SARS 186 00:09:00,640 --> 00:09:04,200 COV-2 SHARE CT IMAGING 187 00:09:04,200 --> 00:09:06,960 CHARACTERISTICS WITH HUMANS 188 00:09:06,960 --> 00:09:09,640 COVID-19 DISEASE. 189 00:09:09,640 --> 00:09:11,320 TOGETHER WITH THIS HAMSTER HIGH 190 00:09:11,320 --> 00:09:13,440 REPLICATION OF THIS VIRUS IN 191 00:09:13,440 --> 00:09:16,880 RESPIRATORY ORGANS, HAMSTERS ARE 192 00:09:16,880 --> 00:09:20,520 USED FOR MODEL FOR COVID-19 193 00:09:20,520 --> 00:09:21,720 STUDIES. 194 00:09:21,720 --> 00:09:24,320 SO WE USED THIS MODEL ANIMAL 195 00:09:24,320 --> 00:09:26,440 MODEL TO EXAMINE POINT MUTATION 196 00:09:26,440 --> 00:09:32,000 ON VIRUS TRANSMISSION. 197 00:09:32,000 --> 00:09:35,280 BETTY CULLIVER FOUND THAT THE 198 00:09:35,280 --> 00:09:39,200 MUTATION AT 614 OF THE S PROTEIN 199 00:09:39,200 --> 00:09:41,840 FROM D TO G WAS RESPONSIBLE FOR 200 00:09:41,840 --> 00:09:44,080 THIS DOMINANCE OF THE DEVICE 201 00:09:44,080 --> 00:09:48,320 LATER ON. 202 00:09:48,320 --> 00:09:53,720 SO IN COLLABORATION WITH THE -- 203 00:09:53,720 --> 00:09:55,160 WE EXAMINED THIS PARTICULAR 204 00:09:55,160 --> 00:09:57,080 MUTATION OF VIRUS TRANSMISSION. 205 00:09:57,080 --> 00:10:03,200 SO RALPH MADE ISOGENIC VIRUS 206 00:10:03,200 --> 00:10:04,920 THAT ONLY DISPOSITION. 207 00:10:04,920 --> 00:10:09,200 SO THE ONE VIRUS CONTAINS 208 00:10:09,200 --> 00:10:10,560 MUTATION DISPOSITION IN THE S 209 00:10:10,560 --> 00:10:14,400 PROTEIN AND THE OTHER VIRUS 210 00:10:14,400 --> 00:10:16,240 CONTAINS G AT THIS POSITION. 211 00:10:16,240 --> 00:10:20,080 SO WE INOCULATED THIS VIRUS INTO 212 00:10:20,080 --> 00:10:23,480 HAMSTER, NEXT DAY WE CO-HOUSED 213 00:10:23,480 --> 00:10:25,600 UNINFECTED ANIMAL IN THE SAME 214 00:10:25,600 --> 00:10:28,480 CAGE BUT THESE TWO ANIMALS 215 00:10:28,480 --> 00:10:29,280 DIVIDED BY MESH. 216 00:10:29,280 --> 00:10:30,440 SO THAT THEY DON'T TOUCH EACH 217 00:10:30,440 --> 00:10:32,040 OTHER. 218 00:10:32,040 --> 00:10:33,960 SO THE ONLY WAY THE VIRUS 219 00:10:33,960 --> 00:10:37,160 TRANSMITTED FROM THIS INOCULATED 220 00:10:37,160 --> 00:10:40,320 HAHAMSTER TO CO-HOUSED UNINFECTD 221 00:10:40,320 --> 00:10:41,840 ANIMAL IS RESPIRATORY DROPLETS 222 00:10:41,840 --> 00:10:45,280 OR AEROSOL. 223 00:10:45,280 --> 00:10:48,960 SO EXAMINE TRANSMISSION ON DAY 4 224 00:10:48,960 --> 00:10:53,120 POST EXPOSURE AND WE DID THIS 225 00:10:53,120 --> 00:10:55,880 WITH EIGHT PAIRS FOR EACH VIRUS 226 00:10:55,880 --> 00:10:59,000 AND FOUND THAT IN BOTH VIRUSES 227 00:10:59,000 --> 00:11:01,920 WE SAW TRANSMISSION IN ALL EIGHT 228 00:11:01,920 --> 00:11:06,480 PAIRS OF ANIMALS. 229 00:11:06,480 --> 00:11:08,520 ON DAY 2 POST EXPOSURE WE DIDN'T 230 00:11:08,520 --> 00:11:10,080 SEE ANY TRANSMISSION WITH THE 231 00:11:10,080 --> 00:11:14,600 VIRUS CONTAINING D AT 232 00:11:14,600 --> 00:11:16,080 DISPOSITION, WE SAW TRANSMISSION 233 00:11:16,080 --> 00:11:19,320 IN FIVE OUT OF EIGHT PAIRS WITH 234 00:11:19,320 --> 00:11:22,680 THE DEVICE CONTAINING GLYCENE AT 235 00:11:22,680 --> 00:11:25,160 THIS POSITION. 236 00:11:25,160 --> 00:11:28,880 SO THIS DATA SHOW THAT S 237 00:11:28,880 --> 00:11:30,680 MUTATION G PROMOWS BETTER 238 00:11:30,680 --> 00:11:33,720 TRANSMISSION. 239 00:11:33,720 --> 00:11:37,960 NOW I WILL TALK ABOUT OMICRON. 240 00:11:37,960 --> 00:11:40,720 SO YOU KNOW DELTA WAVE WAS 241 00:11:40,720 --> 00:11:42,960 REPLACED BY OMICRON BA 1 AND 242 00:11:42,960 --> 00:11:46,000 THEN BA 2. 243 00:11:46,000 --> 00:11:51,360 AND NOW IN UNITED STATES BA 2, 244 00:11:51,360 --> 00:11:52,840 12.1 BECAME DOMINANT AND IN 245 00:11:52,840 --> 00:11:56,480 SOUTH AFRICA, AFTER BA 1 WAVE BA 246 00:11:56,480 --> 00:12:00,360 2 APPEARED AND THEN BA 4 AND BA 247 00:12:00,360 --> 00:12:04,280 5 ARE NOW DOMINATING. 248 00:12:04,280 --> 00:12:06,200 SOME OF THE EUROPEAN COUNTRIES 249 00:12:06,200 --> 00:12:08,320 HAVE BA 4, BA 5 OR BOTH. 250 00:12:08,320 --> 00:12:11,200 BUT TO THEM GOING TO TALK ABOUT 251 00:12:11,200 --> 00:12:14,640 BA TODAY, THE GENUS BA 1. 252 00:12:14,640 --> 00:12:17,160 AND THIS WORK WAS DONE IN THE -- 253 00:12:17,160 --> 00:12:24,600 AS A PART OF THIS SAVE -- MANY 254 00:12:24,600 --> 00:12:26,680 PEOPLE ARE INVOLVED IN THIS 255 00:12:26,680 --> 00:12:30,800 PROJECT, MANY FROM NIH VRC AND 256 00:12:30,800 --> 00:12:33,200 ALSO MY PEOPLE IN MY GROUP IN 257 00:12:33,200 --> 00:12:38,520 TOKYO AND MADISON. 258 00:12:38,520 --> 00:12:43,320 SO WE FIRST TESTED IN MICE USING 259 00:12:43,320 --> 00:12:45,360 FOUR DIFFERENT MOUSE STRAINS, 260 00:12:45,360 --> 00:12:47,720 FOUR DIFFERENT BA 1 CLINICAL 261 00:12:47,720 --> 00:12:48,960 ISOLATES. 262 00:12:48,960 --> 00:12:53,000 AND TESTED DIFFERENT INOCULATING 263 00:12:53,000 --> 00:12:54,840 DOSES. 264 00:12:54,840 --> 00:12:58,600 SO THE DATA COLOR CODED ALL THE 265 00:12:58,600 --> 00:13:01,240 OMICRON DATA BA 1 DATA IN 266 00:13:01,240 --> 00:13:08,160 DIFFERENT COLORS, THE OTHER 267 00:13:08,160 --> 00:13:10,400 VARIANTS PROTO VIRUSES ARE SHOWN 268 00:13:10,400 --> 00:13:10,920 IN BLACK. 269 00:13:10,920 --> 00:13:13,440 IN THE ACE TWO EXPRESSING 270 00:13:13,440 --> 00:13:15,440 TRANSGENIC MICE BA 1 DIDN'T 271 00:13:15,440 --> 00:13:18,480 CAUSE ANY BODY WEIGHT LOSS. 272 00:13:18,480 --> 00:13:21,320 THAT WAS ALSO THE CASE IN 129 273 00:13:21,320 --> 00:13:26,520 MOUSE STRAIN. 274 00:13:26,520 --> 00:13:27,640 AND ALSO B 6 MOUSE. 275 00:13:27,640 --> 00:13:32,960 WE ALSO EXAMINED VIRUS TITER BY 276 00:13:32,960 --> 00:13:36,800 INFECTING THE ANIMALS WITH 277 00:13:36,800 --> 00:13:38,040 ISOLATE. 278 00:13:38,040 --> 00:13:44,160 IN H 2 TRANSGENIC MICE AND ALSO 279 00:13:44,160 --> 00:13:47,160 BALB/C MICE, BA 1 ISOLATE DIDN'T 280 00:13:47,160 --> 00:13:49,320 REPLICATE AS GOOD AS PROTOTYPE 281 00:13:49,320 --> 00:13:53,320 OR BETA VARIANT IN BOTH NASAL 282 00:13:53,320 --> 00:13:57,360 TERMINATE AND LUNGS. 283 00:13:57,360 --> 00:14:04,200 WE THEN EXAMINED HAMSTERS BY 284 00:14:04,200 --> 00:14:06,440 INFECTING HAMSTERS WITH 285 00:14:06,440 --> 00:14:07,040 INFECTIOUS UNITS. 286 00:14:07,040 --> 00:14:09,600 AS YOU CAN SEE THE ANIMALS 287 00:14:09,600 --> 00:14:12,240 INFECTED WITH THE OMICRON BA 1 288 00:14:12,240 --> 00:14:15,440 DIDN'T LOSE BODY WEIGHT UNLIKE 289 00:14:15,440 --> 00:14:20,040 ANIMALS INFECTED WITH OTHER 290 00:14:20,040 --> 00:14:21,120 VIRUS. 291 00:14:21,120 --> 00:14:25,480 WE THEN EXAMINED THE VIRUS TITER 292 00:14:25,480 --> 00:14:29,920 IN RESPIRATORY ORGANS AND YOU 293 00:14:29,920 --> 00:14:32,280 CAN SEE WE DIDN'T SEE ANY 294 00:14:32,280 --> 00:14:34,600 DIFFERENCE BETWEEN DELTA AND 295 00:14:34,600 --> 00:14:37,720 OMICRON IN NASAL TERMINATE BUT 296 00:14:37,720 --> 00:14:39,240 WE SAW SUBSTANTIAL REDUCTION IN 297 00:14:39,240 --> 00:14:41,600 TITER IN OMICRON INFECTED 298 00:14:41,600 --> 00:14:46,080 ANIMALS IN LUNGS. 299 00:14:46,080 --> 00:14:47,680 WE THEN EXAMINED RESPIRATORY 300 00:14:47,680 --> 00:14:55,640 FUNCTION BY USING WHOLE BODY THE 301 00:14:55,640 --> 00:15:00,680 ANIMALS INFECTED WITH OMICRON 302 00:15:00,680 --> 00:15:02,560 WAS SIMPLY INFECTED WITH ANIMALS 303 00:15:02,560 --> 00:15:03,560 IN THESE PARAMETERS DIFFERENT 304 00:15:03,560 --> 00:15:06,360 FROM ANIMALS INFECT WITH DELTA 305 00:15:06,360 --> 00:15:08,200 VARIANT WHICH SHOWS THE 306 00:15:08,200 --> 00:15:11,240 RESPIRATORY FAILURE. 307 00:15:11,240 --> 00:15:13,440 WE ALSO EXAMINED THE LUNG LESION 308 00:15:13,440 --> 00:15:16,480 BY USING MICROCT AND AS YOU CAN 309 00:15:16,480 --> 00:15:19,640 SEE ANIMALS INFECTED WITH DELTA 310 00:15:19,640 --> 00:15:26,480 VARIANT SHOWED MULTIBILL LUNG 311 00:15:26,480 --> 00:15:27,920 CONDENSATIONS AND NO DUALS AND 312 00:15:27,920 --> 00:15:29,680 WE DON'T SEE THOSE IN OMICRON 313 00:15:29,680 --> 00:15:32,640 INFECTED ANIMALS. 314 00:15:32,640 --> 00:15:35,760 WE DID SEE LUNG -- IN ANIMALS 315 00:15:35,760 --> 00:15:39,160 INFECTED WITH OMICRON. 316 00:15:39,160 --> 00:15:41,040 BUT CT SCORING CLEARLY SHOWED 317 00:15:41,040 --> 00:15:44,840 THE DIFFERENCE IN TERMS OF LUNG 318 00:15:44,840 --> 00:15:46,040 LESIONS BETWEEN DELTA AND 319 00:15:46,040 --> 00:15:51,400 OMICRON INFECTED ANIMALS. 320 00:15:51,400 --> 00:15:54,120 THIS CAN BE SEEN BY CT SCAN 321 00:15:54,120 --> 00:15:55,560 MOVIE, YOU CAN SEE MORE SEVERE 322 00:15:55,560 --> 00:15:57,680 LESIONS IN DELTA INFECTED 323 00:15:57,680 --> 00:15:59,040 ANIMALS THAN IN THE OMICRON 324 00:15:59,040 --> 00:16:04,640 INFECTED ANIMALS. 325 00:16:04,640 --> 00:16:06,160 PATHOLOGICALLY WE SAW LESIONS IN 326 00:16:06,160 --> 00:16:09,320 LUNGS OF ANIMALS INFECTED WITH 327 00:16:09,320 --> 00:16:11,040 DELTA BUT NOT IN LUNGS OF 328 00:16:11,040 --> 00:16:15,040 ANIMALS INFECTED WITH OMICRON 329 00:16:15,040 --> 00:16:15,320 VIRUS. 330 00:16:15,320 --> 00:16:18,440 HISTOPATHOLOGICALLY WE SAW EDEMA 331 00:16:18,440 --> 00:16:21,840 IN THE HEMORRHAGE IN ANIMALS 332 00:16:21,840 --> 00:16:25,440 INFECTED DELTA VIRUS BUT 333 00:16:25,440 --> 00:16:26,560 INFLAMMATION OF LUNGS WITH 334 00:16:26,560 --> 00:16:28,080 ANIMALS INFECTED WITH OMICRON IS 335 00:16:28,080 --> 00:16:31,640 LIMITED. 336 00:16:31,640 --> 00:16:33,720 HISTOLOGY SHOWS THIS DIFFERENCE 337 00:16:33,720 --> 00:16:38,800 ON DAY 6 POST INFECTION. 338 00:16:38,800 --> 00:16:42,320 NOW IN SITU HYBRIDIZATION ALSO 339 00:16:42,320 --> 00:16:46,200 SHOW THAT THERE ARE MANY VIRAL 340 00:16:46,200 --> 00:16:50,120 RNA POSITIVE CELLS IN LUNGS. 341 00:16:50,120 --> 00:16:53,440 AND ALSO EPITHELIAL CELLS LINING 342 00:16:53,440 --> 00:16:55,000 BRONCHUS OF ANIMALS INFECTED 343 00:16:55,000 --> 00:16:57,240 WITH DELTA VIRUS. 344 00:16:57,240 --> 00:17:00,200 BUT VERY FEW VIRUS RNA POSITIVE 345 00:17:00,200 --> 00:17:02,160 CELLS IN ANIMALS INFECTED WITH 346 00:17:02,160 --> 00:17:07,120 OMICRON VARIANT. 347 00:17:07,120 --> 00:17:09,560 SO THIS DATA SHOW THE OMICRON BA 348 00:17:09,560 --> 00:17:12,240 1 VARIANT ATTENUATED IN HAMSTERS 349 00:17:12,240 --> 00:17:16,760 AND MICE. 350 00:17:16,760 --> 00:17:22,800 OTHER SHOWED SIMILAR DATA. 351 00:17:22,800 --> 00:17:24,400 NOW I WILL TALK ABOUT COMPARISON 352 00:17:24,400 --> 00:17:29,960 BETWEEN BA 1 AND BA 2. 353 00:17:29,960 --> 00:17:33,880 SO THE GROUP IN JAPAN REPORTED 354 00:17:33,880 --> 00:17:36,600 STUDIES USING THE RECOMBINANT 355 00:17:36,600 --> 00:17:39,680 SARS COV-2 CONTAINING S PROTEIN 356 00:17:39,680 --> 00:17:43,680 FROM DIFFERENT VIRUS. 357 00:17:43,680 --> 00:17:45,760 SO THEY GENERATED RECOMBINANT 358 00:17:45,760 --> 00:17:49,120 VIRUSES POSSESSING Z GENE FROM 359 00:17:49,120 --> 00:17:53,440 DIFFERENT VIRUSES,ER IN THE 360 00:17:53,440 --> 00:17:59,480 BACKGROUND OF WUHAN LIKE SARS 361 00:17:59,480 --> 00:17:59,960 COV-2. 362 00:17:59,960 --> 00:18:02,040 THEY SHOW THE VIRUS CONTAINING 363 00:18:02,040 --> 00:18:04,640 BA 1 S PROTEIN DIDN'T CAUSE BODY 364 00:18:04,640 --> 00:18:06,200 WEIGHT LOSS, THIS IS IN HAMSTERS 365 00:18:06,200 --> 00:18:09,280 BUT THE VIRUS CONTAINING BA 2 S 366 00:18:09,280 --> 00:18:13,200 PROTEIN CAUSED BODY WEIGHT LOSS. 367 00:18:13,200 --> 00:18:16,240 ALSO THE AMOUNT OF VIRAL RNA IN 368 00:18:16,240 --> 00:18:20,320 LUNGS IS HIGHER WITH VIRUS 369 00:18:20,320 --> 00:18:23,560 CONTAINING BA 2 S PROTEIN 370 00:18:23,560 --> 00:18:25,240 COMPARED WITH VIRUS CONTAINING 371 00:18:25,240 --> 00:18:31,000 BA 1 S PROTEIN. 372 00:18:31,000 --> 00:18:34,520 THEY ALSO EXAMINE PATHOLOGY AND 373 00:18:34,520 --> 00:18:36,440 FOUND THAT VIRUS CONTAINING BA 2 374 00:18:36,440 --> 00:18:40,520 S PROTEIN CAUSE MORE SEVERE 375 00:18:40,520 --> 00:18:43,400 PATHOLOGICAL CHANGES. 376 00:18:43,400 --> 00:18:48,360 WE ALSO EXAMINE THE RESPIRATORY 377 00:18:48,360 --> 00:18:53,440 FUNCTION BY USING WHO BODY 378 00:18:53,440 --> 00:18:54,880 PLESTHOGRAPHY, THE VIRUS 379 00:18:54,880 --> 00:18:58,520 CONTAINING BA 2 PROTEIN CAUSED 380 00:18:58,520 --> 00:18:59,200 RESPIRATORY FAILURE. 381 00:18:59,200 --> 00:19:01,640 SO BASED ON THIS DATA THEY 382 00:19:01,640 --> 00:19:03,520 CONCLUDED THE VIRUS CONTAINING 383 00:19:03,520 --> 00:19:06,440 THE BA 2 S PROTEIN IS MORE 384 00:19:06,440 --> 00:19:07,720 PATHOGENIC THAN THE VIRUS 385 00:19:07,720 --> 00:19:13,680 CONTAINING BA 1 S PROTEIN. 386 00:19:13,680 --> 00:19:16,560 SO WE EXAMINED THE PATHOGENICITY 387 00:19:16,560 --> 00:19:22,600 AND COMPARED BETWEEN BA 1 AND BA 388 00:19:22,600 --> 00:19:25,680 2 USING THE CLINICAL ISOLATES. 389 00:19:25,680 --> 00:19:27,760 AND THIS WORK WAS DONE IN 390 00:19:27,760 --> 00:19:31,800 COLLABORATION WITH MANY PEOPLE. 391 00:19:31,800 --> 00:19:33,360 THE ONE AT UTAH STATE UNIVERSITY 392 00:19:33,360 --> 00:19:35,640 WHO PROVIDED US WITH TRANSGENIC 393 00:19:35,640 --> 00:19:39,920 HAMSTERS EXPRESSING HUMAN H 2 394 00:19:39,920 --> 00:19:40,320 PROTEIN. 395 00:19:40,320 --> 00:19:47,040 AND THE GROUP AT THE NIH VRC AND 396 00:19:47,040 --> 00:19:49,680 AUBREY GORDON, UNIVERSITY OF 397 00:19:49,680 --> 00:19:52,000 MICHIGAN AND MIKE DIAMOND, 398 00:19:52,000 --> 00:19:52,760 WASHINGTON UNIVERSITY. 399 00:19:52,760 --> 00:19:57,880 SO WE COMPARED VIRUS REPLICATION 400 00:19:57,880 --> 00:20:08,560 OF BA 1 AND BA 2 ICE RATES IN 401 00:20:08,560 --> 00:20:10,000 BALBC AND THEY REPLICATE TO 402 00:20:10,000 --> 00:20:13,240 SIMILAR EXTENT IN H NASAL 403 00:20:13,240 --> 00:20:15,240 DETERMINANT AND ON DAY FIVE WE 404 00:20:15,240 --> 00:20:17,600 SAW HIGHER REPLICATION WITH BA 1 405 00:20:17,600 --> 00:20:21,880 COMPARED WITH BA 2. 406 00:20:21,880 --> 00:20:25,080 ON DAY 2 THE TITLERS ARE SIMILAR 407 00:20:25,080 --> 00:20:27,040 THOUGH STATISTICALLY SLIGHTLY 408 00:20:27,040 --> 00:20:29,520 DIFFERENT. 409 00:20:29,520 --> 00:20:32,960 IN SITU HYBRIDIZATION AND 410 00:20:32,960 --> 00:20:33,680 HISTOPATHOLOGY SHOW KNOW 411 00:20:33,680 --> 00:20:35,840 DIFFERENCE BETWEEN BA 1 AND BA 2 412 00:20:35,840 --> 00:20:40,400 INFECTED ANIMALS. 413 00:20:40,400 --> 00:20:43,640 WE THEN EXAMINE BODY WEIGHT 414 00:20:43,640 --> 00:20:45,800 CHANGE UPON INFECTION OF 415 00:20:45,800 --> 00:20:48,040 HAMSTERS WITH 10 TO THIRD OR 10 416 00:20:48,040 --> 00:20:50,000 TO FIFTH INFECTIOUS UNITS AND WE 417 00:20:50,000 --> 00:20:51,440 DID NOT SEE ANY DIFFERENCE 418 00:20:51,440 --> 00:20:56,840 BETWEEN BA 1 AND BA 2. 419 00:20:56,840 --> 00:21:00,720 THE RESPIRATORY FUNCTIONS AS 420 00:21:00,720 --> 00:21:06,120 MEASURED BY WHOLE BODY 421 00:21:06,120 --> 00:21:08,200 PLETHSMAGRAPHY, WE DID NOT SEE 422 00:21:08,200 --> 00:21:09,760 ENDARCH BETWEEN BA 1 AND BA 2, 423 00:21:09,760 --> 00:21:12,960 BOTH ANIMALS ARE SIMILAR TO 424 00:21:12,960 --> 00:21:16,280 INFECTED. 425 00:21:16,280 --> 00:21:21,640 VIRUS REPLICATION ON DAY 3 WITH 426 00:21:21,640 --> 00:21:24,640 TWO DOSES 10 TO THIRD OR 10 TO 427 00:21:24,640 --> 00:21:28,680 FIFTH IN NASAL TERM FANTASTIC 428 00:21:28,680 --> 00:21:32,120 WITH SIMILAR TITER BA 1 AND 2 429 00:21:32,120 --> 00:21:34,800 WITH THE 10 TO 3RD INTEGRATION 430 00:21:34,800 --> 00:21:38,240 WE SAW BA 1 WAS HIGHER IN BA 2 431 00:21:38,240 --> 00:21:42,640 IN LUNG TITER. 432 00:21:42,640 --> 00:21:44,880 WITH 10 TO 5TH INTEGRATION WE 433 00:21:44,880 --> 00:21:48,480 DIDN'T SEE ANY DIFFERENCE. 434 00:21:48,480 --> 00:21:50,320 THE HISTOPATHOLOGICALLY AND IN 435 00:21:50,320 --> 00:21:52,560 SITU HYBRIDIZATION SHOW NO 436 00:21:52,560 --> 00:21:54,000 DIFFERENCE BETWEEN BA 1 AND 2 IN 437 00:21:54,000 --> 00:21:56,200 HAMSTER. 438 00:21:56,200 --> 00:22:06,720 TUSHNATE WE THEN INVOLVED BA 1 439 00:22:09,120 --> 00:22:12,200 AND BA 2 MIX, WE INTENDED TO MIX 440 00:22:12,200 --> 00:22:14,000 ONE TO ONE BUT WHEN WE ANALYZE 441 00:22:14,000 --> 00:22:18,440 BY SEQUENCING WE FOUND RATIO WAS 442 00:22:18,440 --> 00:22:21,480 3 TO 7. 443 00:22:21,480 --> 00:22:26,440 WE THEN EXAMINED THE VIRUS RATIO 444 00:22:26,440 --> 00:22:31,240 DAY 2 AND DAY 4 IN BOTH TIME 445 00:22:31,240 --> 00:22:34,560 POINT BA 1 BECAME MORE PREVALENT 446 00:22:34,560 --> 00:22:37,440 IN NASAL TURBINATE AND LUNG IN 447 00:22:37,440 --> 00:22:41,160 MOST ANIMALS. 448 00:22:41,160 --> 00:22:44,320 SO THIS DATA SHOW THAT FOR ALL 449 00:22:44,320 --> 00:22:47,720 PARAMETERS TESTED, BA 1 AND BA 2 450 00:22:47,720 --> 00:22:53,200 SHOW SIMILAR PATHOGENICITY. 451 00:22:53,200 --> 00:22:57,920 THIS IS SIMILAR TO WHAT IS 452 00:22:57,920 --> 00:23:00,640 HAPPENING IN CLINICAL SETTING 453 00:23:00,640 --> 00:23:02,280 SHOWN IN THIS PRE-PRINT, 454 00:23:02,280 --> 00:23:04,160 CLINICAL PROFILE OF ILLNESS 455 00:23:04,160 --> 00:23:05,800 REMAINS SIMILAR BETWEEN BA 2 AND 456 00:23:05,800 --> 00:23:06,600 BA 1. 457 00:23:06,600 --> 00:23:09,360 BUT RECENT PRE-PRINT SHOWS 458 00:23:09,360 --> 00:23:10,440 SOMETHING DIFFERENT. 459 00:23:10,440 --> 00:23:13,440 IN THIS PREPRINT DESCRIBE BA 2 460 00:23:13,440 --> 00:23:16,800 WAS ASSOCIATED WITH REPORTING 461 00:23:16,800 --> 00:23:19,280 MORE SYMPTOMS WITH GREATER 462 00:23:19,280 --> 00:23:20,360 DISRUPTION TO DAILY ACTIVITIES 463 00:23:20,360 --> 00:23:28,320 THAN BA 1. 464 00:23:28,320 --> 00:23:31,960 NOW, BA 1, BA 11 AND BA 2 DIFFER 465 00:23:31,960 --> 00:23:33,560 IN THEIR S PROTEIN. 466 00:23:33,560 --> 00:23:37,440 THIS PICTURE WAS I BORROWED IT 467 00:23:37,440 --> 00:23:41,000 FROM DAVID HO'S PAPER PUBLISHED 468 00:23:41,000 --> 00:23:45,720 IN NATURE. 469 00:23:45,720 --> 00:23:49,920 THE INSTITUTION SHOWN IN BLACK, 470 00:23:49,920 --> 00:23:51,280 SUBSTITUTION COMMON BETWEEN BA 1 471 00:23:51,280 --> 00:23:54,920 AND BA 2. 472 00:23:54,920 --> 00:23:56,960 IMMUNOASSAY CHANGES SHOWN IN 473 00:23:56,960 --> 00:23:58,880 GREEN RESIDUE SPECIFIC FOR BA 1 474 00:23:58,880 --> 00:24:01,600 AND BA 11. 475 00:24:01,600 --> 00:24:05,520 AND BA 1.1 CONTAINS ONE 476 00:24:05,520 --> 00:24:08,480 ADDITIONAL MUTATION OF 346 FROM 477 00:24:08,480 --> 00:24:13,280 ARGININE TO LYSINE. 478 00:24:13,280 --> 00:24:15,280 CHANGES SHOWN IN BLUE SPECIFIC 479 00:24:15,280 --> 00:24:20,360 FOR BA 2. 480 00:24:20,360 --> 00:24:23,800 MANY PAPERS DESCRIBE 481 00:24:23,800 --> 00:24:24,800 NEUTRALIZATION OF OMICRON 482 00:24:24,800 --> 00:24:29,920 VARIANTS BY USING THE SERA FROM 483 00:24:29,920 --> 00:24:31,280 PATIENTS OR VACCINEES. 484 00:24:31,280 --> 00:24:34,360 WE ALSO DID SIMILAR EXPERIMENTS, 485 00:24:34,360 --> 00:24:39,120 THESE ARE THE SERA FROM VACCINE 486 00:24:39,120 --> 00:24:44,520 CANDIDATES, VACCINEES AND 487 00:24:44,520 --> 00:24:47,640 INFECTED AND THEN VACCINATED AND 488 00:24:47,640 --> 00:24:50,480 ONE MONTH TO THREE MONTHS AND 489 00:24:50,480 --> 00:24:53,720 SIX MONTHS AFTER VACCINATION. 490 00:24:53,720 --> 00:24:56,360 AGAINST WUHAN DELTA AND THREE 491 00:24:56,360 --> 00:24:58,400 DIFFERENT OMICRON VARIANTS. 492 00:24:58,400 --> 00:25:01,120 THESE ARE THE SERA FROM PEOPLE 493 00:25:01,120 --> 00:25:03,760 WHO RECEIVED THREE PFIZER 494 00:25:03,760 --> 00:25:04,360 VACCINATIONS. 495 00:25:04,360 --> 00:25:08,400 AND THESE SERA FROM THE mRNA 496 00:25:08,400 --> 00:25:09,400 VACCINES AND THEN OMICRON 497 00:25:09,400 --> 00:25:11,600 INFECTION. 498 00:25:11,600 --> 00:25:13,800 IN ALL CASES, THE -- YOU SEE 499 00:25:13,800 --> 00:25:19,560 HIGHER TITER AGAINST WUHAN NEXT 500 00:25:19,560 --> 00:25:23,560 DELTA, AND THEN OMICRON. 501 00:25:23,560 --> 00:25:25,880 WE ALSO DID SIMILAR 502 00:25:25,880 --> 00:25:27,080 NEUTRALIZATION ASSAY USING 503 00:25:27,080 --> 00:25:27,840 HAMSTERS HERE. 504 00:25:27,840 --> 00:25:31,520 SO HAMSTERS WERE INFECTED WITH 505 00:25:31,520 --> 00:25:34,360 ANCESTRAL VIRUS DELTA BA 1 BA 506 00:25:34,360 --> 00:25:36,600 1.1 OR BA 1. 507 00:25:36,600 --> 00:25:38,400 1 -- BA 2 AND TESTED 508 00:25:38,400 --> 00:25:39,640 AGAINST SAME VIRUSES. 509 00:25:39,640 --> 00:25:42,520 AND AS YOU SEE, BA 1 INFECTED 510 00:25:42,520 --> 00:25:45,080 HAMSTERS HERE REACT WELL WITH BA 511 00:25:45,080 --> 00:25:49,160 2 BUT LESS SO AGAINST OTHER 512 00:25:49,160 --> 00:25:53,160 OMICRON VARIANTS AND BA 1 OR BA 513 00:25:53,160 --> 00:25:57,480 1.1 INFECTED HAMSTERS REACT WITH 514 00:25:57,480 --> 00:26:02,320 ALL THREE BA OMICRON VARIANTS, 515 00:26:02,320 --> 00:26:06,200 LESS SO WUHAN AND DELTA. 516 00:26:06,200 --> 00:26:07,440 REVERSE THE SAME WITH ANCESTRAL 517 00:26:07,440 --> 00:26:11,440 OR DELTA INFECTED HAMSTERS HERE. 518 00:26:11,440 --> 00:26:14,600 THESE RESULTS ARE QUITE SIMILAR 519 00:26:14,600 --> 00:26:15,480 TO THOSE WE SEE WITH HUMANS 520 00:26:15,480 --> 00:26:18,840 HERE. 521 00:26:18,840 --> 00:26:22,040 SO HAMSTER SERA ARE QUITE USEFUL 522 00:26:22,040 --> 00:26:25,800 TO ANTIGEN ANALYSIS. 523 00:26:25,800 --> 00:26:29,680 IN FACT (INDISCERNIBLE) USED 524 00:26:29,680 --> 00:26:32,880 THIS HAMSTER SCALE TO GENERAL 525 00:26:32,880 --> 00:26:35,640 RATE ANTIGENIC OF SARS COV-2 S 526 00:26:35,640 --> 00:26:36,120 PROTEIN. 527 00:26:36,120 --> 00:26:38,560 THIS TECHNIQUE WAS ORIGINALLY 528 00:26:38,560 --> 00:26:43,520 DEVELOPED BY DERRICK SMITH AND 529 00:26:43,520 --> 00:26:48,720 (INDISCERNIBLE) AND USED THIS 530 00:26:48,720 --> 00:26:50,000 METHOD TO PERFORM ANGIOGENIC 531 00:26:50,000 --> 00:26:55,880 ANALYSIS. 532 00:26:55,880 --> 00:26:57,560 THE ONE GREEK UNIT HERE 533 00:26:57,560 --> 00:26:59,480 CORRESPONDS TO TWO FULL DILUTION 534 00:26:59,480 --> 00:27:01,480 OF SERA. 535 00:27:01,480 --> 00:27:05,080 AND THIS -- THIS IS THE 6 536 00:27:05,080 --> 00:27:06,120 PROTEIN G ONE OF THE ANCESTRAL 537 00:27:06,120 --> 00:27:06,680 VIRUS. 538 00:27:06,680 --> 00:27:08,760 THESE ARE THE OTHER VARIANTS. 539 00:27:08,760 --> 00:27:11,480 AND AS YOU CAN SEE, BA 1 AND BA 540 00:27:11,480 --> 00:27:15,960 2 ARE LOCATED HERE WHICH IS FAR 541 00:27:15,960 --> 00:27:19,880 AWAY FROM ANCESTRAL VIRUS. 542 00:27:19,880 --> 00:27:24,040 THESE TRANSGENIC RESPONSE TO 543 00:27:24,040 --> 00:27:26,360 ANGIOGENIC DIFFERENCES. 544 00:27:26,360 --> 00:27:29,800 IN MANY GROUP DESCRIBED 545 00:27:29,800 --> 00:27:32,120 NEUTRALIZATION AGAINST THESE 546 00:27:32,120 --> 00:27:36,880 OMICRON VARIANTS, WE ALSO DID 547 00:27:36,880 --> 00:27:42,120 EXPERIMENTS USING OMICRON AND 548 00:27:42,120 --> 00:27:44,800 FOUND THAT ANTIBODIES REACT -- 549 00:27:44,800 --> 00:27:46,680 DID NOT NEUTRALIZE ANY OF THE 550 00:27:46,680 --> 00:27:49,280 OMICRON VARIANTS BA 1 BA 1.1, BA 551 00:27:49,280 --> 00:27:54,280 2. 552 00:27:54,280 --> 00:27:57,520 ONE OF THE NOT NEUTRALIZED BA 1 553 00:27:57,520 --> 00:28:01,720 AND BA 1.1 BUT DID NEUTRALIZE BA 554 00:28:01,720 --> 00:28:04,320 2 THOUGH TO LESSER EXTENT 555 00:28:04,320 --> 00:28:06,240 COMPARED WITH THE ORIGINAL WUHAN 556 00:28:06,240 --> 00:28:08,640 VIRUS. 557 00:28:08,640 --> 00:28:11,840 THE TRANSGENIC ANTIBODIES ONE 558 00:28:11,840 --> 00:28:16,280 NEUTRALIZED BA 2 AS WELL AS 559 00:28:16,280 --> 00:28:17,320 WUHAN VIRUS. 560 00:28:17,320 --> 00:28:21,880 SO THIS COCKTAIL REACTED WITH BA 561 00:28:21,880 --> 00:28:23,360 2 ONLY 15 FOLD LESS COMPARED 562 00:28:23,360 --> 00:28:29,200 WITH THE WUHAN LIKE VIRUS. 563 00:28:29,200 --> 00:28:33,040 S 309 WHICH IS A PRECURSOR OF 564 00:28:33,040 --> 00:28:34,320 (INAUDIBLE) HAD REACTIVITY 565 00:28:34,320 --> 00:28:37,520 AGAINST BA 1 AND BA 1.1 WITH 566 00:28:37,520 --> 00:28:40,440 PROTEIN FOLD REDUCTION. 567 00:28:40,440 --> 00:28:43,680 50 FOLD REDUCTION AGAINST THE BA 568 00:28:43,680 --> 00:28:49,800 .2 VARIANT. 569 00:28:49,800 --> 00:28:53,080 SO DIFFERS AMONG OMICRON 570 00:28:53,080 --> 00:28:55,960 VARIANTS, WE THEN EXAMINED THE 571 00:28:55,960 --> 00:28:57,880 EFFICACY OF THESE MONOCLONAL 572 00:28:57,880 --> 00:28:59,400 ANTIBODIES AGAINST OMICRON 573 00:28:59,400 --> 00:29:01,720 VARIANTS IN HAMSTERS. 574 00:29:01,720 --> 00:29:04,120 SO WE FIRST INFECTED HAMSTERS 575 00:29:04,120 --> 00:29:06,600 WITH 10 TO THE THIRD INFECTIOUS 576 00:29:06,600 --> 00:29:08,720 UNITS AND THEN WITH THESE 577 00:29:08,720 --> 00:29:12,880 VIRUSES AND THEN NEXT DAY WE 578 00:29:12,880 --> 00:29:16,160 INOCULATED FIVE CHILDREN WITH 579 00:29:16,160 --> 00:29:17,520 ANTIBODY IN (INAUDIBLE) AND CAM 580 00:29:17,520 --> 00:29:26,120 MINUTED VIRUS TITER ON DAY 4. 581 00:29:26,120 --> 00:29:31,000 AGAINST THE 614G VIRUS 582 00:29:31,000 --> 00:29:33,200 (INAUDIBLE) PRECURSOR IS 309 583 00:29:33,200 --> 00:29:35,800 REGENERON COCKTAIL AND ASTRA 584 00:29:35,800 --> 00:29:37,280 ZENECA COCKTAIL REDUCE VIRUS 585 00:29:37,280 --> 00:29:41,240 TITER IN LUNGS AND REGENERON 586 00:29:41,240 --> 00:29:48,640 COCKTAIL AND ASTRA ZENECA 587 00:29:48,640 --> 00:29:51,840 COCKTAIL AGAINST BA 1 ASTRA 588 00:29:51,840 --> 00:29:53,320 ZENECA REDUCED TITER IN THE 589 00:29:53,320 --> 00:29:57,600 LUNGS, NONE REDUCED TITTER IN 590 00:29:57,600 --> 00:29:58,720 NASAL TURBINATE. 591 00:29:58,720 --> 00:30:00,480 AGAINST BA 2 ALL THREE ANIMALS 592 00:30:00,480 --> 00:30:03,040 -- ANTIBODIES REDUCED TITER 593 00:30:03,040 --> 00:30:07,000 THOUGH THE WE DID NOT SEE 594 00:30:07,000 --> 00:30:08,800 STATISTICAL DIFFERENCE WITH THIS 595 00:30:08,800 --> 00:30:09,760 REGENERON ANTIBODY COCKTAIL IN 596 00:30:09,760 --> 00:30:10,800 LUNG TITERS. 597 00:30:10,800 --> 00:30:15,240 BUT NONE OF THESE ANTIBODIES 598 00:30:15,240 --> 00:30:20,800 REDUCE TITER IN NASAL TURBINATE. 599 00:30:20,800 --> 00:30:24,840 BECAUSE OF THIS ANGIOGENIC 600 00:30:24,840 --> 00:30:28,920 DIFFERENCES, AS YOU ALL KNOW, 601 00:30:28,920 --> 00:30:30,880 THE VACCINE EFFECTIVENESS OF 602 00:30:30,880 --> 00:30:32,160 MRNA AGAINST OMICRON IS 603 00:30:32,160 --> 00:30:36,120 REDUCED. 604 00:30:36,120 --> 00:30:38,560 BUT THE QUESTION HERE IS THAT 605 00:30:38,560 --> 00:30:41,680 OMICRON INFECTION IN HUMANS, DUE 606 00:30:41,680 --> 00:30:44,000 TO IMMUNITY WANING OR ANGIOGENIC 607 00:30:44,000 --> 00:30:46,960 CHANGES IN INFECTED VIRUS OR 608 00:30:46,960 --> 00:30:48,320 BOTH. 609 00:30:48,320 --> 00:30:50,240 WE ATTEMPTED TO ANSWER THIS 610 00:30:50,240 --> 00:30:53,040 QUESTION UNDER CONTROLLED 611 00:30:53,040 --> 00:30:54,240 CONDITIONS, USING OMICRON 612 00:30:54,240 --> 00:30:56,320 VARIANT AND ISOLATE THAT ARE NO 613 00:30:56,320 --> 00:31:00,040 LONGER CIRCULATING IN NATURE. 614 00:31:00,040 --> 00:31:02,560 THAT IS WUHAN LIKE ISOLATE AND 615 00:31:02,560 --> 00:31:06,760 NOT ISOLATE WITH ONLY 614G SPIKE 616 00:31:06,760 --> 00:31:11,360 MUTATION IN HAMSTER MODEL. 617 00:31:11,360 --> 00:31:16,720 SO WE INFECTED HAMSTERS WITH 618 00:31:16,720 --> 00:31:21,160 614G VIRUS OR WUHAN LIKE VIRUS 619 00:31:21,160 --> 00:31:23,240 AND 7 MONTHS OR 22 MONTHS AFTER 620 00:31:23,240 --> 00:31:24,800 INFECTION WE CHALLENGE THOSE 621 00:31:24,800 --> 00:31:28,320 ANIMALS WITH EITHER OMICRON OR 622 00:31:28,320 --> 00:31:30,280 614G VIRUS. 623 00:31:30,280 --> 00:31:33,720 ON DAY 4 POST INFECTION WE 624 00:31:33,720 --> 00:31:36,880 EXAMINED THE VIRUS SITE. 625 00:31:36,880 --> 00:31:40,320 IN AGE MATCHED NAIVE ANIMALS, 626 00:31:40,320 --> 00:31:42,240 THESE 614G VIRUS REPLICATED WELL 627 00:31:42,240 --> 00:31:45,600 AS EXPECTED. 628 00:31:45,600 --> 00:31:47,800 BUT IN THE ANIMALS PREVIOUSLY 629 00:31:47,800 --> 00:31:51,440 INFECTD WITH 614G SEVEN MONTHS 630 00:31:51,440 --> 00:31:53,760 AGO WUHAN LIKE VIRUS 22 MONTHS 631 00:31:53,760 --> 00:31:57,240 AGO, IN THESE ANIMALS 614G VIRUS 632 00:31:57,240 --> 00:31:59,640 DIDN'T REPLICATE. 633 00:31:59,640 --> 00:32:03,680 SO IMMUNITY AGAINST ANGIOGENIC 634 00:32:03,680 --> 00:32:07,360 SIMILAR VIRUS LASTS FOR AT LEAST 635 00:32:07,360 --> 00:32:08,080 22 MONTHS. 636 00:32:08,080 --> 00:32:10,400 AFTER INFECTION. 637 00:32:10,400 --> 00:32:14,400 IN AGE MATCHED ANIMALS OMICRON 638 00:32:14,400 --> 00:32:16,600 VIRUS REPLICATED WELL, THOUGH TO 639 00:32:16,600 --> 00:32:19,640 LESSER EXTENT AS I DESCRIBED. 640 00:32:19,640 --> 00:32:21,640 BUT IN ANIMALS PREVIOUSLY 641 00:32:21,640 --> 00:32:25,160 INFECTED WITH 614G OR WUHAN LIKE 642 00:32:25,160 --> 00:32:27,360 VIRUS, OMICRON DIDN'T REPLICATE 643 00:32:27,360 --> 00:32:29,480 IN LUNGS, BUT DID REPLICATE IN 644 00:32:29,480 --> 00:32:35,360 NASAL TURBINATE. 645 00:32:35,360 --> 00:32:37,160 100 FOLD REDUCTION COMPARED WITH 646 00:32:37,160 --> 00:32:40,400 NAIVE ANIMALS. 647 00:32:40,400 --> 00:32:41,480 THIS DIFFERENCE IN REPLICATION 648 00:32:41,480 --> 00:32:45,040 HERE IS DUE TO THE ANGIOGENIC 649 00:32:45,040 --> 00:32:46,360 DIFFERENCE BETWEEN THE PROTOTYPE 650 00:32:46,360 --> 00:32:50,000 VIRUS AND OMICRON. 651 00:32:50,000 --> 00:32:52,400 SO PRODUCTIVITY IMMUNITY LASTS 652 00:32:52,400 --> 00:32:54,600 FOR 22 MONTHS AFTER THE ANTIGEN 653 00:32:54,600 --> 00:32:56,920 IS -- WHEN THE ANTIGEN IS 654 00:32:56,920 --> 00:32:58,400 DETERMINED VIRUS IS SIMILAR TO 655 00:32:58,400 --> 00:33:01,400 THAT OF THE INITIAL VIRUS LISTED 656 00:33:01,400 --> 00:33:04,840 -- AT LEAST IN HAMSTERS. 657 00:33:04,840 --> 00:33:07,560 WE THEN EXAMINED PROTECTION OF 658 00:33:07,560 --> 00:33:09,120 WEAKER INFECTION AGAINST THE 659 00:33:09,120 --> 00:33:12,200 VACCINATED ANIMALS. 660 00:33:12,200 --> 00:33:15,360 AND SO WE VACCINATED HAMSTERS 661 00:33:15,360 --> 00:33:18,800 WITH MODERNA mRNA ONCE OR 662 00:33:18,800 --> 00:33:20,720 TWICE AND SEVEN MONTHS LATER WE 663 00:33:20,720 --> 00:33:22,480 CHALLENGED THESE ANIMALS WITH 664 00:33:22,480 --> 00:33:24,560 OMICRON OR PROTOTYPE VIRUS. 665 00:33:24,560 --> 00:33:31,000 THEN EXAMINED THE DATA ON DAY 4. 666 00:33:31,000 --> 00:33:33,480 AS YOU CAN SEE THE VIRUS TITER 667 00:33:33,480 --> 00:33:36,880 IS REDUCED ABOUT SIX LOG UNITS 668 00:33:36,880 --> 00:33:38,800 IN LUNGS, IN ANIMALS CHALLENGED 669 00:33:38,800 --> 00:33:40,840 WITH THE PROTOTYPE VIRUS. 670 00:33:40,840 --> 00:33:45,520 AFTER TWO DOSES OF VACCINATION. 671 00:33:45,520 --> 00:33:46,520 TWO AND A HALF LOG UNIT 672 00:33:46,520 --> 00:33:50,520 REDUCTION IN NASAL TURBINATE IN 673 00:33:50,520 --> 00:33:53,360 ANIMALS VACCINATED TWICE. 674 00:33:53,360 --> 00:33:54,720 SO THAT IS AGAINST PROTOTYPE 675 00:33:54,720 --> 00:33:56,200 CHALLENGE. 676 00:33:56,200 --> 00:33:58,960 BUT IN ANIMALS CHALLENGED WITH 677 00:33:58,960 --> 00:34:02,240 OMICRON WE SAW ONLY TWO LOG 678 00:34:02,240 --> 00:34:03,800 REDUCTION IN ANIMALS VACCINATED 679 00:34:03,800 --> 00:34:06,560 TWICE. 680 00:34:06,560 --> 00:34:08,320 IN LUNGS AND ONE AND A HALF LOG 681 00:34:08,320 --> 00:34:14,120 UNIT REDUCTION IN ANIMALS 682 00:34:14,120 --> 00:34:16,240 RECEIVING TWO DOSES VACCINATION. 683 00:34:16,240 --> 00:34:18,680 SO THIS DIFFERENCE IS DUE TO THE 684 00:34:18,680 --> 00:34:22,800 ANGIOGENIC DIFFERENCES. 685 00:34:22,800 --> 00:34:25,240 SO THE PROTOTYPE VACCINE IS 686 00:34:25,240 --> 00:34:27,040 PROTECTIVE AGAINST THE PROTOTYPE 687 00:34:27,040 --> 00:34:30,680 VACCINE IS PROTECTIVE AGAINST 688 00:34:30,680 --> 00:34:32,040 PROTOTYPE VIRUS EVEN SEVEN 689 00:34:32,040 --> 00:34:33,800 MONTHS ASP VACCINATION. 690 00:34:33,800 --> 00:34:38,200 BUT LESS PROTECTIVE AGAINST 691 00:34:38,200 --> 00:34:39,720 ANTIGENICALLY DIFFERENT OMICRON 692 00:34:39,720 --> 00:34:43,800 VARIANT. 693 00:34:43,800 --> 00:34:47,680 WE TESTED LOW MOLECULAR WEIGHT 694 00:34:47,680 --> 00:34:50,360 COMPOUNDS, REMDESIVIR, 695 00:34:50,360 --> 00:34:54,960 POLYMERASE INHIBITOR, 696 00:34:54,960 --> 00:34:56,640 NIRMATRELVIR PROTEASE INHIBITOR. 697 00:34:56,640 --> 00:34:59,360 ALL THREE WORK GAINS BA 1, BA 698 00:34:59,360 --> 00:35:03,200 1.1 BA 2 QUITE WELL IN VITRO. 699 00:35:03,200 --> 00:35:06,760 WE THEN TESTED THE EFFICACY OF 700 00:35:06,760 --> 00:35:14,640 THIS COMPOUNDS AND S 217622 701 00:35:14,640 --> 00:35:19,200 PRODUCE BY -- COMPANY CLINICAL 702 00:35:19,200 --> 00:35:23,440 TRIAL IN HAMSTER MODEL. 703 00:35:23,440 --> 00:35:25,680 WE INFECTED HAMSTERS WITH BA 1 704 00:35:25,680 --> 00:35:29,440 OR BA 2 WITH 10 TO THIRD 705 00:35:29,440 --> 00:35:30,360 INFECTIOUS UNITS. 706 00:35:30,360 --> 00:35:33,080 NEXT DAY WE STARTED TREATMENT, 707 00:35:33,080 --> 00:35:35,360 TWICE A DAY ORALLY. 708 00:35:35,360 --> 00:35:37,880 WITH THESE COMPOUNDS AND THEN ON 709 00:35:37,880 --> 00:35:39,240 DAY 4 WE EXAMINED THE VIRUS 710 00:35:39,240 --> 00:35:43,000 TITER. 711 00:35:43,000 --> 00:35:51,240 NOW AGAINST BA 121-7622 WORKED 712 00:35:51,240 --> 00:35:53,480 WELL REDUCED VIRUS TITER TO ZERO 713 00:35:53,480 --> 00:35:57,400 IN LUNGS. 714 00:35:57,400 --> 00:35:59,640 AND 217622 ALSO REDUCED VIRUS 715 00:35:59,640 --> 00:36:02,440 TITER IN NASAL TURBINATE. 716 00:36:02,440 --> 00:36:07,160 AGAINST BA 2 ALL THREE COMPOUNDS 717 00:36:07,160 --> 00:36:08,360 REDUCED VIRUS TITERS 718 00:36:08,360 --> 00:36:12,600 SUBSTANTIALLY ALMOST TO ZERO AND 719 00:36:12,600 --> 00:36:17,680 TO UNDETECTABLE LEVEL AND 720 00:36:17,680 --> 00:36:19,160 NIRMATRELVIR ALSO REDUCED VIRUS 721 00:36:19,160 --> 00:36:21,240 TITER IN NASAL TURBINATE. 722 00:36:21,240 --> 00:36:22,880 SO SMALL MOLECULE DRUGS 723 00:36:22,880 --> 00:36:25,600 CURRENTLY APPROVED OR IN 724 00:36:25,600 --> 00:36:28,080 CLINICAL TRIALS EFFECTIVELY 725 00:36:28,080 --> 00:36:30,360 INHIBIT REPLICATION OMICRON BA 1 726 00:36:30,360 --> 00:36:36,680 AND BA 2 IN LUNGS OF HAMSTERS. 727 00:36:36,680 --> 00:36:38,120 NOW I'M GOING TO TALK ABOUT 728 00:36:38,120 --> 00:36:41,280 IMAGING. 729 00:36:41,280 --> 00:36:42,960 THE AIM OF THE STUDIES TO 730 00:36:42,960 --> 00:36:46,280 UNDERSTAND COVID-19 PATHOGENESIS 731 00:36:46,280 --> 00:36:48,200 BY USING IMAGING SYSTEMS AND 732 00:36:48,200 --> 00:36:54,640 THIS WORK WAS DONE BY 733 00:36:54,640 --> 00:36:56,200 (INDISCERNIBLE). 734 00:36:56,200 --> 00:36:59,080 IN THAT GROUP WE ESTABLISHED 735 00:36:59,080 --> 00:37:00,880 MULTIPLE DIFFERENT IMAGING 736 00:37:00,880 --> 00:37:02,680 SYSTEMS FOR INFLUENZA RESEARCH. 737 00:37:02,680 --> 00:37:04,840 TODAY I WILL TALK ABOUT TWO 738 00:37:04,840 --> 00:37:07,840 PHOTON MICROSCOPY. 739 00:37:07,840 --> 00:37:10,400 SO THIS IS HOW WE DO THIS 740 00:37:10,400 --> 00:37:11,520 EXPERIMENT. 741 00:37:11,520 --> 00:37:15,480 WE HIT MICE WITH LASER AND 742 00:37:15,480 --> 00:37:19,360 PROJECT IMAGE ON MONITOR. 743 00:37:19,360 --> 00:37:21,880 MISO IN HEATED STAGE AND TO LOOK 744 00:37:21,880 --> 00:37:25,840 INSIDE THE LUNGS WE PLACED 745 00:37:25,840 --> 00:37:30,840 WINDOW CONNECTED TO VACUUM PUMP, 746 00:37:30,840 --> 00:37:33,360 WE HAVE ANESTHESIA MACHINE, 747 00:37:33,360 --> 00:37:35,040 MECHANICAL VENTILATOR, TO 748 00:37:35,040 --> 00:37:36,880 PREVENT CONTAMINATION OF THIS 749 00:37:36,880 --> 00:37:41,320 DEVICE WE PUT HEPA FILTER AND WE 750 00:37:41,320 --> 00:37:43,480 PLACED THE ENTIRE SYSTEM IN BSL 751 00:37:43,480 --> 00:37:45,560 3 SO OPERATOR WITH PROTECTIVE 752 00:37:45,560 --> 00:37:47,960 GEAR. 753 00:37:47,960 --> 00:37:52,160 THIS IS HOW WE DO THIS 754 00:37:52,160 --> 00:37:52,880 EXPERIMENT. 755 00:37:52,880 --> 00:37:56,760 WE PERFORM TRACHEOTOMY AND 756 00:37:56,760 --> 00:37:58,960 INSERTED CANNELLA. 757 00:37:58,960 --> 00:38:04,560 MAKE INCISION, AND EXPOSE LUNG. 758 00:38:04,560 --> 00:38:07,480 LUNG MOVES LIKE THIS. 759 00:38:07,480 --> 00:38:10,480 SO WE NEED TO STOP THE MOVEMENT 760 00:38:10,480 --> 00:38:12,280 TO DO THE EXPERIMENT. 761 00:38:12,280 --> 00:38:17,360 SO WE PLACE THE THORACIC WINDOW 762 00:38:17,360 --> 00:38:20,600 THROUGH THE VACUUM SO THAT 763 00:38:20,600 --> 00:38:24,240 SURFACE OF THE LUNG BECAME 764 00:38:24,240 --> 00:38:28,320 STABILIZED. 765 00:38:28,320 --> 00:38:32,200 THEN PLACE OBJECTIVE LENS ON TOP 766 00:38:32,200 --> 00:38:38,480 OF THIS LUNG AND THEN OBSERVE. 767 00:38:38,480 --> 00:38:40,400 SO THIS IS HOW OUR SET UP IN BSL 768 00:38:40,400 --> 00:38:42,400 3 AND THIS IS DR. (INAUDIBLE) 769 00:38:42,400 --> 00:38:48,160 PERFORMING THIS EXPERIMENT. 770 00:38:48,160 --> 00:38:49,720 SO WE USE WOE ESTABLISH THE 771 00:38:49,720 --> 00:38:52,760 SYSTEM FOR INFLUENZA FIRST AND 772 00:38:52,760 --> 00:38:56,280 THEN NOW USE THE SYSTEM FOR SARS 773 00:38:56,280 --> 00:38:56,800 COV-2. 774 00:38:56,800 --> 00:39:01,080 SO WE INFECTED H 2 EXPRESSING 775 00:39:01,080 --> 00:39:04,560 TRANSGENIC MICE WITH SARS COV-2 776 00:39:04,560 --> 00:39:05,680 EXPRESSING VENUS FLUORESCENT 777 00:39:05,680 --> 00:39:06,440 PROTEIN. 778 00:39:06,440 --> 00:39:10,640 WHICH IS GENERATED BY DR. 779 00:39:10,640 --> 00:39:14,600 KAMITANI AT GUNMA UNIVERSITY. 780 00:39:14,600 --> 00:39:19,920 WE INJECTED DEXTRAN TO LOOK AT 781 00:39:19,920 --> 00:39:22,280 BLOOD FLOW ANTIBODY TO 782 00:39:22,280 --> 00:39:25,520 NEUTROPHIL, ANTIBODY 2 PLATELET. 783 00:39:25,520 --> 00:39:27,760 AND THIS IS LUNG OF ANIMALS 784 00:39:27,760 --> 00:39:29,280 INFECT WITH SARS COV-2 785 00:39:29,280 --> 00:39:32,080 EXPRESSING VENUS PROTEIN. 786 00:39:32,080 --> 00:39:38,440 AND GRAY INDICATES BLACK FILL, 787 00:39:38,440 --> 00:39:40,120 GREEN INDICATES VIRUS INDICATED 788 00:39:40,120 --> 00:39:45,040 WITH CELLS, RED INDICATES 789 00:39:45,040 --> 00:39:48,080 NEUTROPHIL, CIENNE INDICATES 790 00:39:48,080 --> 00:39:49,040 PLATELETS. 791 00:39:49,040 --> 00:39:51,680 TYPE 1 AND TYPE 2 PNEUMOCYTES 792 00:39:51,680 --> 00:39:55,680 ARE INFECTED IN THE TRANSGENIC 793 00:39:55,680 --> 00:40:00,600 ANIMALS. 794 00:40:00,600 --> 00:40:04,080 NOW WE OBSERVE -- WE FOUND THAT 795 00:40:04,080 --> 00:40:06,760 ENHANCED -- WE FOUND ENHANCED 796 00:40:06,760 --> 00:40:08,880 VASCULAR PERMEABILITY IN ANIMALS 797 00:40:08,880 --> 00:40:12,280 INFECTED WITH SARS COV-2, YOU 798 00:40:12,280 --> 00:40:15,640 SEE CLOUDNESS OF ALVEOLI IN THE 799 00:40:15,640 --> 00:40:17,440 VIRUS INFECTED ANIMALS. 800 00:40:17,440 --> 00:40:20,000 THIS ACTUALLY CAN BE QUANTIFIED 801 00:40:20,000 --> 00:40:23,400 BY EXAMINING THE AMOUNT OF 802 00:40:23,400 --> 00:40:26,880 DEXTRAN LEADING TO ALVEOLI. 803 00:40:26,880 --> 00:40:32,000 YOU SEE INCREASE VASCULAR 804 00:40:32,000 --> 00:40:33,920 PERMEABILITY AS THE INFECTION 805 00:40:33,920 --> 00:40:35,960 PROGRESSED. 806 00:40:35,960 --> 00:40:38,320 WE ALSO EXAMINED NUMBER OF 807 00:40:38,320 --> 00:40:40,840 NEUTROPHILS INCREASE OVER TIME. 808 00:40:40,840 --> 00:40:42,560 NEUTROPHILS SHOWN IN RED IN 809 00:40:42,560 --> 00:40:47,640 NAIVE ANIMALS MOVE FAST BUT 810 00:40:47,640 --> 00:40:49,280 NEUTROPHILS IN THE VIRUS 811 00:40:49,280 --> 00:40:51,200 INFECTED ANIMALS, THE NUMBER IS 812 00:40:51,200 --> 00:40:52,880 INCREASE AS SHOWN HERE. 813 00:40:52,880 --> 00:40:57,240 ALSO THEY DON'T MOVE. 814 00:40:57,240 --> 00:41:01,320 THO MOVEMENT CAN BE TRACED. 815 00:41:01,320 --> 00:41:03,440 THEN QUANTIFIED. 816 00:41:03,440 --> 00:41:06,240 AS YOU CAN SEE, NEUTROPHILS IN 817 00:41:06,240 --> 00:41:08,080 NAIVE ANIMALS MOVE QUICKLY, AS 818 00:41:08,080 --> 00:41:10,520 SHOWN IN THIS DIAGRAM. 819 00:41:10,520 --> 00:41:12,560 BUT NEUTROPHIL IN VIRUS INFECTED 820 00:41:12,560 --> 00:41:13,760 ANIMALS ON DAY 6, THEY DON'T 821 00:41:13,760 --> 00:41:20,600 MOVE. 822 00:41:20,600 --> 00:41:22,840 WE ALSO FOUND THAT MICROTHROMBUS 823 00:41:22,840 --> 00:41:24,440 LIKE STRUCTURE COMPOSED OF 824 00:41:24,440 --> 00:41:27,480 PLATELET SHOWN IN SIGH YEN AND 825 00:41:27,480 --> 00:41:29,640 NEUTROPHILS SHOWN IN RED. 826 00:41:29,640 --> 00:41:32,840 AND NEUTROPHILS THAT REMAIN IN 827 00:41:32,840 --> 00:41:36,440 THE BLOOD VESSEL MAY CAUSE 828 00:41:36,440 --> 00:41:37,600 THROMBUS FORMATION SEEN IN 829 00:41:37,600 --> 00:41:42,800 COVID-19 PNEUMONIA. 830 00:41:42,800 --> 00:41:46,480 WE ALSO FOUND FORMATION 831 00:41:46,480 --> 00:41:47,720 MICROTHROMBI IN CAPILLARIES OF 832 00:41:47,720 --> 00:41:53,480 VIRUS INFECTED LUNGS. 833 00:41:53,480 --> 00:41:58,160 SO WE EXTRACTED PLATELET SIGNALS 834 00:41:58,160 --> 00:42:01,080 FROM ORIGINAL IMAGE, IDENTIFIED 835 00:42:01,080 --> 00:42:02,960 PLATELETS BASED OR FLUORESCENT 836 00:42:02,960 --> 00:42:04,680 INTENSITY AND SIZE AND MADE 837 00:42:04,680 --> 00:42:07,360 HISTOGRAM. 838 00:42:07,360 --> 00:42:11,480 AS YOU CAN SEE SINGLE PLATELET 839 00:42:11,480 --> 00:42:15,480 PROPORTION OF SINGLE PLATELET 840 00:42:15,480 --> 00:42:18,120 REDUCED AFTER INFECTION AND 841 00:42:18,120 --> 00:42:21,800 AGGRAVATED PLATELETS THROMBUS 842 00:42:21,800 --> 00:42:22,800 INCREASE AFTER INFECTION OVER 843 00:42:22,800 --> 00:42:27,600 TIME. 844 00:42:27,600 --> 00:42:29,760 NOW, LUNG IS ONE O OF THE PLACES 845 00:42:29,760 --> 00:42:33,760 WHERE PLATELETS ARE GENERATED 846 00:42:33,760 --> 00:42:35,840 FROM MEGA KARYOCYTES. 847 00:42:35,840 --> 00:42:40,720 IN UNINFECTED LUNGS, MEGA 848 00:42:40,720 --> 00:42:44,480 KARYOCYTES ARE TORN APART BY FOR 849 00:42:44,480 --> 00:42:48,680 PLATELET GENERATION BY STEADY 850 00:42:48,680 --> 00:42:49,560 BLOOD FLOW. 851 00:42:49,560 --> 00:42:51,680 OR CARRIED AWAY BY THE BLOOD 852 00:42:51,680 --> 00:42:52,680 FLOW. 853 00:42:52,680 --> 00:42:56,760 SO YOU CAN SEE THE GENERATION OF 854 00:42:56,760 --> 00:42:58,600 PLATELETS FROM MEGAKARYOCYTES IN 855 00:42:58,600 --> 00:43:01,840 LUNGS. 856 00:43:01,840 --> 00:43:06,280 SO THIS IS IN NAIVE ANIMAL. 857 00:43:06,280 --> 00:43:08,880 IN VIRUS INFECTED LUNGS, BLOOD 858 00:43:08,880 --> 00:43:13,520 FLOW IS CONGESTED AND 859 00:43:13,520 --> 00:43:14,560 MEGAKARYOCYTES ARE MORE EASILY 860 00:43:14,560 --> 00:43:15,800 TRAPPED IN BLOOD VESSELS WHICH 861 00:43:15,800 --> 00:43:21,440 MAY BE A CAUSE OF EMBOLISM. 862 00:43:21,440 --> 00:43:26,520 SO WE NEXT EXAMINE BLOOD FLOW BY 863 00:43:26,520 --> 00:43:29,080 INFECTING H 2 EXPRESSING 864 00:43:29,080 --> 00:43:30,760 TRANSGENIC MICE WITH SARS COV-2 865 00:43:30,760 --> 00:43:33,520 AND SIX DAYS POST INFECTION WE 866 00:43:33,520 --> 00:43:37,040 INTRODUCED FLUORESCENT LABELED 867 00:43:37,040 --> 00:43:38,240 ERYTHROCYTES. 868 00:43:38,240 --> 00:43:40,200 THEN PERFORM 2 PHOTON 869 00:43:40,200 --> 00:43:41,880 MICROSCOPY. 870 00:43:41,880 --> 00:43:46,440 WE THEN TOOK SHOT EVERY 1.2 871 00:43:46,440 --> 00:43:47,560 SECONDS AND THEN OVERLAID. 872 00:43:47,560 --> 00:43:50,040 FUL SO THIS IS THE NAIVE ANIMAL, 873 00:43:50,040 --> 00:43:54,600 THIS IS SARS COV-2 INFECTED 874 00:43:54,600 --> 00:43:57,720 ANIMALS AND YOU SEE THE 875 00:43:57,720 --> 00:44:01,960 ACCUMULATION OF ERYTHROCYTES 876 00:44:01,960 --> 00:44:04,760 PASS THROUGH THESE CAPILLARIES. 877 00:44:04,760 --> 00:44:07,520 SO WHEN ERYTHROCYTES PASS 878 00:44:07,520 --> 00:44:09,760 THROUGH YOU SEE RED. 879 00:44:09,760 --> 00:44:11,440 IF THEY DON'T GO THROUGH YOU 880 00:44:11,440 --> 00:44:14,040 DON'T SEE ANY RED. 881 00:44:14,040 --> 00:44:16,720 BLUE INDICATES BLOOD FLOW. 882 00:44:16,720 --> 00:44:20,160 SO IN NAIVE ANIMALS AS YOU 883 00:44:20,160 --> 00:44:21,840 EXPECT, ERYTHROCYTES PASS 884 00:44:21,840 --> 00:44:24,720 THROUGH CAPILLARIES, BUT IN SARS 885 00:44:24,720 --> 00:44:27,520 COV-2 INFECTED ANIMALS THERE ARE 886 00:44:27,520 --> 00:44:31,240 AREA YOU DON'T SEE ANY 887 00:44:31,240 --> 00:44:35,200 ERYTHROCYTES GOING THROUGH. 888 00:44:35,200 --> 00:44:38,040 SO IN LUNGS INFECTED WITH SARS 889 00:44:38,040 --> 00:44:39,800 COV-2 CAPILLARIES AVAILABLE FOR 890 00:44:39,800 --> 00:44:45,240 OXYGEN EXCHANGE ARE LIMITED. 891 00:44:45,240 --> 00:44:49,880 SO 2 PHOTON LASER MICROSCOPY 892 00:44:49,880 --> 00:44:52,240 REVEAL FUNCTIONAL FAILURE OF 893 00:44:52,240 --> 00:44:54,720 LUNG ARE DUE TO THROMBI 894 00:44:54,720 --> 00:44:56,000 FORMATION IN MICE INFECT WITH 895 00:44:56,000 --> 00:44:58,440 SARS COV-2. 896 00:44:58,440 --> 00:45:00,680 SO LAST FEW MINUTES I'M GOING TO 897 00:45:00,680 --> 00:45:02,920 TALK ABOUT STABILITY OF SARS 898 00:45:02,920 --> 00:45:08,040 COV-2 IN AEROSOL. 899 00:45:08,040 --> 00:45:09,600 TWO GROUPS PREVIOUSLY DESCRIBED 900 00:45:09,600 --> 00:45:13,360 THE STABILITY OF SARS COV-2 IN 901 00:45:13,360 --> 00:45:19,640 AEROSOL USING ROTATING DRUMS 902 00:45:19,640 --> 00:45:23,080 SIMILAR TO (INAUDIBLE). 903 00:45:23,080 --> 00:45:25,080 DESCRIBE THAT HALF LIFE OF SARS 904 00:45:25,080 --> 00:45:27,920 COV-2 IN AEROSOL IS 905 00:45:27,920 --> 00:45:30,120 APPROXIMATELY 1.1 HOUR. 906 00:45:30,120 --> 00:45:32,680 ANOTHER GROUP REPORTED THAT 907 00:45:32,680 --> 00:45:34,120 INFECTIVITY OF SARS COV-2 908 00:45:34,120 --> 00:45:38,160 REMAINS FOR UP TO 16 HOURS IN 909 00:45:38,160 --> 00:45:39,200 AEROSOL. 910 00:45:39,200 --> 00:45:40,440 SO THEY COULDN'T DETERMINE HALF 911 00:45:40,440 --> 00:45:45,200 LIFE. 912 00:45:45,200 --> 00:45:46,960 SO WE WANTED TO TIME THE 913 00:45:46,960 --> 00:45:48,840 STABILITY OF SARS COV-2 IN 914 00:45:48,840 --> 00:45:51,240 AEROSOL IN SETTING DIFFERENT 915 00:45:51,240 --> 00:45:54,680 FROM THAT USE IN THE PREVIOUS 916 00:45:54,680 --> 00:45:55,080 STUDIES. 917 00:45:55,080 --> 00:46:00,200 SO WE SET UP A SYSTEM IN BSL 3 918 00:46:00,200 --> 00:46:04,520 FACILITY. 919 00:46:04,520 --> 00:46:06,960 HERE WE HAVE NEBULIZER CHAMBER 920 00:46:06,960 --> 00:46:09,040 AND HUMIDIFIER TO CONTROL THE 921 00:46:09,040 --> 00:46:12,360 HUMIDITY OF THIS CHAMBER AND 922 00:46:12,360 --> 00:46:15,720 THEN AIR SAMPLER. 923 00:46:15,720 --> 00:46:18,000 NEBULIZER REDUCES DROPLETS IN 924 00:46:18,000 --> 00:46:21,240 AEROSOL CONTAINING SARS COV-2 AT 925 00:46:21,240 --> 00:46:24,840 THE SPEED OF 2-METER PER SECOND. 926 00:46:24,840 --> 00:46:27,440 THE OTHER PARTICLE SIZE RELEASED 927 00:46:27,440 --> 00:46:29,080 AT TIME OF -- RELEASE OF THIS 928 00:46:29,080 --> 00:46:30,760 DEVICE AT THE TIME OF THE 929 00:46:30,760 --> 00:46:35,440 RERELEASE IS 5.5 MICROLITER. 930 00:46:35,440 --> 00:46:41,160 THE CHAMBER WAS THEN PRODUCED BY 931 00:46:41,160 --> 00:46:43,400 THE (INDISCERNIBLE) UNIVERSITY 932 00:46:43,400 --> 00:46:48,120 BY USING NEBULIZER WE SPRAYED 933 00:46:48,120 --> 00:46:51,520 AEROSOL CONTAINING SARS COV-2 934 00:46:51,520 --> 00:46:56,720 FOR FIVE MINUTES, FOR FIRST ONE 935 00:46:56,720 --> 00:46:57,280 MINUTE THERE WERE TOO MANY 936 00:46:57,280 --> 00:46:58,720 PARTICLES TO COUNT BUT THEN WE 937 00:46:58,720 --> 00:47:00,720 WERE ABLE TO COUNT NUMBER OF 938 00:47:00,720 --> 00:47:03,080 AEROSOLS. 939 00:47:03,080 --> 00:47:06,080 NUMBER OF TOTAL PARTICLES 940 00:47:06,080 --> 00:47:07,280 RELATIVELY HIGH FOR FIRST 24 941 00:47:07,280 --> 00:47:11,560 HOURS. 942 00:47:11,560 --> 00:47:13,760 THERE WAS A DIFFERENCE IN NUMBER 943 00:47:13,760 --> 00:47:16,560 OF DIFFERENT SIZE PARTICLES OVER 944 00:47:16,560 --> 00:47:17,320 TIME. 945 00:47:17,320 --> 00:47:21,480 THE PARTICLES WITH DIAMETER FIVE 946 00:47:21,480 --> 00:47:26,800 MICROMETER REDUCED IN FIRST FEW 947 00:47:26,800 --> 00:47:27,000 HOURS. 948 00:47:27,000 --> 00:47:32,160 AND PARTICLES WITH DIAMETER OF 2 949 00:47:32,160 --> 00:47:33,320 MICROMETER INCREASE AND THEN 950 00:47:33,320 --> 00:47:35,880 DECREASED. 951 00:47:35,880 --> 00:47:39,080 PARTICLE WITH DIAMETER OF 1 952 00:47:39,080 --> 00:47:40,760 MICROMETER GRADUALLY INCREASED 953 00:47:40,760 --> 00:47:43,280 AND THEN REDUCED. 954 00:47:43,280 --> 00:47:46,800 PARTICLES WITH DIAMETER OF .3 OR 955 00:47:46,800 --> 00:47:48,560 .5 MICROMETER GRADUALLY 956 00:47:48,560 --> 00:47:51,080 INCREASE. 957 00:47:51,080 --> 00:47:53,800 SO WHAT THIS INDICATES IS THAT 958 00:47:53,800 --> 00:47:56,960 WATER IN AEROSOLS HAVE UPGRADES 959 00:47:56,960 --> 00:47:58,360 AND SIZE OF AEROSOLS CHANGED 960 00:47:58,360 --> 00:47:59,480 OVER TIME BUT THE TOTAL NUMBER 961 00:47:59,480 --> 00:48:03,480 OF AEROSOLS REMAINS LARGELY THE 962 00:48:03,480 --> 00:48:08,640 SAME. 963 00:48:08,640 --> 00:48:11,160 AIR SAMPLER, THE OPENING OF THE 964 00:48:11,160 --> 00:48:13,120 AIR SAMPLER WE HAVE GELATIN 965 00:48:13,120 --> 00:48:16,120 FILTER WHICH COLLECTS VIRUS 966 00:48:16,120 --> 00:48:19,320 PARTICLE DURING CERTAIN OF AIR. 967 00:48:19,320 --> 00:48:22,680 THIS FILTER DISSOLVES IN CULTURE 968 00:48:22,680 --> 00:48:24,640 MEDIA EASILY TO SO THAT WE CAN 969 00:48:24,640 --> 00:48:26,480 DETERMINE AMOUNT OF INFECTIOUS 970 00:48:26,480 --> 00:48:27,760 VIRUS IN THE VIRAL RNA COPY 971 00:48:27,760 --> 00:48:31,080 NUMBER. 972 00:48:31,080 --> 00:48:33,640 SO HERE WE HAVE NEBULIZER TO 973 00:48:33,640 --> 00:48:35,520 SPRAY AEROSOL CONTAINING SARS 974 00:48:35,520 --> 00:48:38,520 COV-2 INTO CHAMBER AND THEN AT 975 00:48:38,520 --> 00:48:43,560 DIFFERENT TIME POINTS, WE GET 976 00:48:43,560 --> 00:48:48,960 THE RECOVER VIRUS BY USING AIR 977 00:48:48,960 --> 00:48:49,360 SAMPLER. 978 00:48:49,360 --> 00:48:51,280 THE RESULTS ARE SHOWN IN THIS 979 00:48:51,280 --> 00:48:51,880 SLIDE. 980 00:48:51,880 --> 00:48:54,920 BLUE INDICATES INFECTIOUS VIRUS 981 00:48:54,920 --> 00:48:55,960 TITER. 982 00:48:55,960 --> 00:48:57,640 GOLD INDICATES VIRUS RNA COPY 983 00:48:57,640 --> 00:49:00,800 NUMBER. 984 00:49:00,800 --> 00:49:02,880 THE FIRST LUNG WE SEE REDUCTION 985 00:49:02,880 --> 00:49:06,640 IN TITER MOST LIKELY LARGE 986 00:49:06,640 --> 00:49:09,760 PARTICLE SETTLING. 987 00:49:09,760 --> 00:49:12,560 BUT AFTER ONE HOUR WE SEE TITER 988 00:49:12,560 --> 00:49:18,200 REMAIN RELATIVELY STABLE. 989 00:49:18,200 --> 00:49:20,440 WE CAN DETERMINE PERCENTAGE OF 990 00:49:20,440 --> 00:49:23,240 INFECTIOUS VIRUS DIVIDING VIRUS 991 00:49:23,240 --> 00:49:25,920 TITER BY VIRUS RNA COPY NUMBER. 992 00:49:25,920 --> 00:49:28,640 WE FOUND THAT THE PERCENTAGE OF 993 00:49:28,640 --> 00:49:32,320 INFECTIOUS SARS COV-2 IN AEROSOL 994 00:49:32,320 --> 00:49:37,280 REMAINED CONSTANT FOR THREE 995 00:49:37,280 --> 00:49:37,720 HOURS. 996 00:49:37,720 --> 00:49:43,400 SARS COV-2 IN AEROSOL SETTING IS 997 00:49:43,400 --> 00:49:44,760 STAIN SO GOOD INDOOR VENTILATION 998 00:49:44,760 --> 00:49:50,240 IS IMPORTANT. 999 00:49:50,240 --> 00:49:51,880 IN SUMMARY SARS COV-2 1000 00:49:51,880 --> 00:49:54,240 NATURALIZED SHOWED SIMILAR 1001 00:49:54,240 --> 00:49:56,440 PATHOGENICITY TO BA 1 NATURAL 1002 00:49:56,440 --> 00:49:58,840 ISOLATES IN MICE AND HAMSTERS 1003 00:49:58,840 --> 00:50:01,200 AND CURRENTLY IMPROVED 1004 00:50:01,200 --> 00:50:03,680 POLYMERASE AND PROTEASE 1005 00:50:03,680 --> 00:50:05,800 INHIBITORS ARE IMPROVED IN 1006 00:50:05,800 --> 00:50:07,680 OMICRON VARIANTS AND 2 PHOTON 1007 00:50:07,680 --> 00:50:09,760 LASER MICROSCOPY CAN BE USED TO 1008 00:50:09,760 --> 00:50:11,720 ANALYZE SARS COV-2 PATHOGENESIS 1009 00:50:11,720 --> 00:50:13,960 IN MICE AND BECAUSE SARS COVE 26 1010 00:50:13,960 --> 00:50:16,640 IN AEROSOLS IS STABLE, FREQUENT 1011 00:50:16,640 --> 00:50:18,440 VENTILATION IS IMPORTANT. 1012 00:50:18,440 --> 00:50:19,240 THANK YOU VERY MUCH FOR YOUR 1013 00:50:19,240 --> 00:50:27,280 ATTENTION. 1014 00:50:27,280 --> 00:50:31,040 WONDERFUL TALK AS USUAL. 1015 00:50:31,040 --> 00:50:32,920 I'M (INAUDIBLE) AND YOU CAN 1016 00:50:32,920 --> 00:50:35,280 SUBMIT YOUR QUESTIONS TO SEND 1017 00:50:35,280 --> 00:50:37,720 LIVE FEEDBACK BUTTON IN THE 1018 00:50:37,720 --> 00:50:38,520 MIDDLE OF YOUR SCREEN. 1019 00:50:38,520 --> 00:50:39,560 WE HAVE A LOT OF QUESTIONS SO I 1020 00:50:39,560 --> 00:50:43,640 WILL START WITH A FEW OF THEM. 1021 00:50:43,640 --> 00:50:45,200 WHAT DO YOU THINK ANY ANIMAL 1022 00:50:45,200 --> 00:50:48,880 MODEL THAT RECAPITULATES THE 1023 00:50:48,880 --> 00:50:50,840 CARDIAC NEUROLOGICAL SYMPTOMS WE 1024 00:50:50,840 --> 00:50:52,360 SEE IN HUMANS? 1025 00:50:52,360 --> 00:50:57,880 >> RIGHT. 1026 00:50:57,880 --> 00:51:00,560 NONE ANIMAL MODELS -- WILL 1027 00:51:00,560 --> 00:51:01,320 CAPTURE EVERYTHING. 1028 00:51:01,320 --> 00:51:02,920 THE HAMSTERS ARE HIGHLY 1029 00:51:02,920 --> 00:51:05,800 SUSCEPTIBLE SO IT IS A GOOD 1030 00:51:05,800 --> 00:51:09,160 ANIMAL MODEL VIROLOGICAL 1031 00:51:09,160 --> 00:51:09,760 STUDIES. 1032 00:51:09,760 --> 00:51:11,000 AND PROBABLY GOOD ANIMAL MODELS 1033 00:51:11,000 --> 00:51:14,480 FOR PATHOGENESIS IN RESPIRATORY 1034 00:51:14,480 --> 00:51:15,080 ORGANS. 1035 00:51:15,080 --> 00:51:18,040 BUT WHETHER THIS IS GOOD ANIMAL 1036 00:51:18,040 --> 00:51:21,200 MODEL FOR KNOWN RESPIRATORY 1037 00:51:21,200 --> 00:51:25,280 ORGANS REQUIRE MORE STUDIES. 1038 00:51:25,280 --> 00:51:31,120 WE SEE SOME ANTIGEN CELLS IN THE 1039 00:51:31,120 --> 00:51:33,400 HEART SO THERE MAYBE SOME 1040 00:51:33,400 --> 00:51:37,720 REPLICATION IN HEART BUT WHETHER 1041 00:51:37,720 --> 00:51:38,400 THAT CAUSE DISEASE IN -- SIMILAR 1042 00:51:38,400 --> 00:51:41,120 DISEASE IN HAMSTER OR NOT I'M 1043 00:51:41,120 --> 00:51:44,160 NOT SURE. 1044 00:51:44,160 --> 00:51:46,000 >> SINCE YOU HAVE ANIMALS THAT 1045 00:51:46,000 --> 00:51:48,200 YOU HAVE KEPT FOR MANY MONTHS 1046 00:51:48,200 --> 00:51:50,760 POST INFECTION, DO YOU SEE ANY 1047 00:51:50,760 --> 00:51:52,680 OF THOSE ANIMALS THAT MIMIC LONG 1048 00:51:52,680 --> 00:51:57,440 TERM SEQUELAE OR 1049 00:51:57,440 --> 00:51:57,880 (INDISCERNIBLE)? 1050 00:51:57,880 --> 00:51:59,760 >> YES, THAT IS THE QUESTION TO 1051 00:51:59,760 --> 00:52:00,440 ADDRESS. 1052 00:52:00,440 --> 00:52:01,560 WE HAVEN'T DONE ANY EXPERIMENTS. 1053 00:52:01,560 --> 00:52:04,320 I THINK THIS ANIMAL MODEL IS ONE 1054 00:52:04,320 --> 00:52:07,160 OF THE GOOD ONE BECAUSE IT WAS A 1055 00:52:07,160 --> 00:52:09,800 SMALL ANIMAL SO WE CAN KEPT 1056 00:52:09,800 --> 00:52:13,200 ANIMALS LONG TIME EASIER THAN 1057 00:52:13,200 --> 00:52:14,760 MONKEYS SO I THINK WE AND OTHERS 1058 00:52:14,760 --> 00:52:19,360 WILL BE DOING THAT. 1059 00:52:19,360 --> 00:52:21,440 >> ONE QUESTION ASKED ORIGINAL 1060 00:52:21,440 --> 00:52:23,880 SARS COV-2 STRAIN ARE PROTOTYPE 1061 00:52:23,880 --> 00:52:27,400 STRAIN EVOKES HYPERIMMUNE 1062 00:52:27,400 --> 00:52:28,600 HYPERCOAGULOPATHY IN PATIENTS, 1063 00:52:28,600 --> 00:52:28,840 US RIGHT? 1064 00:52:28,840 --> 00:52:30,920 BUT IN YOUR ANIMAL MODELS THAT 1065 00:52:30,920 --> 00:52:32,760 SHOWED LESS SEVERITY DO YOU 1066 00:52:32,760 --> 00:52:33,840 THINK THE OMICRON VARIANT CAUSE 1067 00:52:33,840 --> 00:52:42,480 A LESS HYPERIMMUNETY? 1068 00:52:42,480 --> 00:52:48,480 >> IN HAMSTER MODELS OMICRON IS 1069 00:52:48,480 --> 00:52:51,640 (INAUDIBLE) AND MAYBE SO IN 1070 00:52:51,640 --> 00:52:52,160 HUMANS. 1071 00:52:52,160 --> 00:52:55,000 BUT ATTENUATION IN HAMSTERS WITH 1072 00:52:55,000 --> 00:52:58,320 OMICRON MAYBE MORE PROMINENT 1073 00:52:58,320 --> 00:53:02,040 COMPARED WITH IN HUMANS. 1074 00:53:02,040 --> 00:53:04,320 SO BECAUSE OF THAT I AM NOT SURE 1075 00:53:04,320 --> 00:53:04,960 WHETHER I CAN ADDRESS THE 1076 00:53:04,960 --> 00:53:10,520 QUESTION. 1077 00:53:10,520 --> 00:53:12,080 ANTIGEN MULTIPLE STUDIES ON 1078 00:53:12,080 --> 00:53:13,360 DRUGS AND VACCINES AND 1079 00:53:13,360 --> 00:53:18,320 THERAPEUTIC ANTIBODIES, IN 1080 00:53:18,320 --> 00:53:20,960 HAMSTERS DO YOU SEE SARS COV-2 1081 00:53:20,960 --> 00:53:22,680 DRIFTING OR ESCAPING TREATMENT 1082 00:53:22,680 --> 00:53:25,280 OVER TIME AS OBSERVED IN 1083 00:53:25,280 --> 00:53:25,920 (INDISCERNIBLE)? 1084 00:53:25,920 --> 00:53:29,480 >> CLEARLY MONOCLONAL ANTIBODIES 1085 00:53:29,480 --> 00:53:30,960 BECAUSE OF MUTATION IN S 1086 00:53:30,960 --> 00:53:34,240 PROTEINS AND VARIANTS, SOME OF 1087 00:53:34,240 --> 00:53:35,240 THE VARIANTS ESCAPE FROM THIS 1088 00:53:35,240 --> 00:53:38,360 TREATMENT. 1089 00:53:38,360 --> 00:53:39,920 BUT AT LEAST WITH THIS NORMAL 1090 00:53:39,920 --> 00:53:42,480 MOLECULAR WEIGHT COMPOUNDS, THEY 1091 00:53:42,480 --> 00:53:44,560 ARE EFFICACIOUS AGAINST THE 1092 00:53:44,560 --> 00:53:46,080 VIRUSES WE TESTED INCLUDING ALL 1093 00:53:46,080 --> 00:53:50,640 THE VARIANTS WE TESTED. 1094 00:53:50,640 --> 00:53:52,360 SO THAT IS THAT. 1095 00:53:52,360 --> 00:53:55,840 >> AND IN THE STUDIES THAT YOU 1096 00:53:55,840 --> 00:53:57,280 MENTION IN THE (INAUDIBLE) 1097 00:53:57,280 --> 00:53:59,400 STUDIES HAVE YOU TRIED THAT WITH 1098 00:53:59,400 --> 00:54:00,280 OMICRON VARIANTS? 1099 00:54:00,280 --> 00:54:01,200 ARE THERE DIFFERENCES? 1100 00:54:01,200 --> 00:54:04,200 >> WE HAVEN'T DONE THAT. 1101 00:54:04,200 --> 00:54:06,040 WE WANT TO DO THAT EXPERIMENT. 1102 00:54:06,040 --> 00:54:09,880 >> I THINK ONE OF THE QUESTIONS 1103 00:54:09,880 --> 00:54:12,520 THAT IS WE ARE NOW SEEING BREAK 1104 00:54:12,520 --> 00:54:13,560 THROUGH INFECTIONS FOLLOWING 1105 00:54:13,560 --> 00:54:14,160 VACCINATION. 1106 00:54:14,160 --> 00:54:15,520 SO CAN ANY OF THESE ANIMAL 1107 00:54:15,520 --> 00:54:17,600 MODELS INFORM US THE DEVELOPMENT 1108 00:54:17,600 --> 00:54:19,480 OF MORE EFFECTIVE OR MORE 1109 00:54:19,480 --> 00:54:20,080 AVAILABLE RESPONSES FOR 1110 00:54:20,080 --> 00:54:21,280 VACCINES? 1111 00:54:21,280 --> 00:54:25,520 >> I THINK AS I DESCRIBED, IT IS 1112 00:54:25,520 --> 00:54:27,680 ALWAYS IMPORTANT TO SEE 1113 00:54:27,680 --> 00:54:28,840 DIFFERENT ANIMAL MODELS OR 1114 00:54:28,840 --> 00:54:32,160 DIFFERENT ANIMAL SPECIES CAN 1115 00:54:32,160 --> 00:54:33,400 RECOGNIZE PROTEIN, IN THIS CASE 1116 00:54:33,400 --> 00:54:34,280 S PROTEIN, THE SAME WAY AS 1117 00:54:34,280 --> 00:54:35,680 HUMANS. 1118 00:54:35,680 --> 00:54:40,920 AND AS I SHOWED, HAMSTERS SERA 1119 00:54:40,920 --> 00:54:43,160 GIVE SIMILAR REACTIVITY AGAINST 1120 00:54:43,160 --> 00:54:46,920 DIFFERENT VARIANTS AS WE SEE IN 1121 00:54:46,920 --> 00:54:47,520 HUMANS. 1122 00:54:47,520 --> 00:54:49,080 SO IN -- BECAUSE OF THAT I THINK 1123 00:54:49,080 --> 00:54:51,080 HAMSTER MAYBE A GOOD ANIMAL 1124 00:54:51,080 --> 00:54:53,320 MODEL FOR STUDYING VACCINE 1125 00:54:53,320 --> 00:54:54,280 EFFICACY AGAINST DIFFERENT 1126 00:54:54,280 --> 00:54:55,200 VARIANTS. 1127 00:54:55,200 --> 00:55:00,760 >> IN YOUR ANIMAL MODEL IF I 1128 00:55:00,760 --> 00:55:02,400 READ THAT BA 1 VERSUS BA 2 YOU 1129 00:55:02,400 --> 00:55:08,760 SAW BA 1 WAS OVERCOMING BA 2 IN 1130 00:55:08,760 --> 00:55:09,400 THE MIXTURE EXPERIMENT, RIGHT? 1131 00:55:09,400 --> 00:55:09,800 >> YES. 1132 00:55:09,800 --> 00:55:11,800 >> BUT NOW IN HUMANS AT LEAST WE 1133 00:55:11,800 --> 00:55:17,360 ARE SEEING THAT BA 2 OR BA 2, OR 1134 00:55:17,360 --> 00:55:18,920 BA 1.1 -- WHAT IS GOING ON AND 1135 00:55:18,920 --> 00:55:20,800 HOW CAN WE UNDERSTAND THAT? 1136 00:55:20,800 --> 00:55:25,800 >> I THINK THAT IS DUE TO 1137 00:55:25,800 --> 00:55:26,960 IMMUNITY HUMAN. 1138 00:55:26,960 --> 00:55:30,920 HERE WE USE NAIVE ANIMALS SO THE 1139 00:55:30,920 --> 00:55:32,080 NEXT OBVIOUS EXPLAIN IS TO BE 1140 00:55:32,080 --> 00:55:33,560 DONE IN ANIMALS PREVIOUSLY 1141 00:55:33,560 --> 00:55:38,480 VACCINATED OR INFECTED. 1142 00:55:38,480 --> 00:55:41,840 >> AND DO YOU SEE TRANSMISSION 1143 00:55:41,840 --> 00:55:43,840 IN THE TWO BARE I CAN'T WANT, BA 1144 00:55:43,840 --> 00:55:47,040 2 VERSUS BA 21.1? 1145 00:55:47,040 --> 00:55:49,240 >> THAT IS A REALLY IMPORTANT 1146 00:55:49,240 --> 00:55:53,280 QUESTION AND NOT ONLY OUR GROUP 1147 00:55:53,280 --> 00:55:54,400 BUT ALSO SEVERAL OTHER GROUP 1148 00:55:54,400 --> 00:55:55,880 TESTED HAMSTER FOR AIRBORNE 1149 00:55:55,880 --> 00:55:57,480 TRANSMISSION. 1150 00:55:57,480 --> 00:55:59,040 AND WE DON'T SEE AIRBORNE 1151 00:55:59,040 --> 00:56:01,120 TRANSMISSION IN OMICRON VARIANT 1152 00:56:01,120 --> 00:56:02,200 IN HAMSTERS. 1153 00:56:02,200 --> 00:56:05,280 SO AT THE MOMENT THERE IS NO 1154 00:56:05,280 --> 00:56:07,160 GOOD AIRBORNE TRANSMISSION 1155 00:56:07,160 --> 00:56:08,160 ANIMAL MODEL SYSTEM. 1156 00:56:08,160 --> 00:56:13,080 WE DO SEE DIRECT TRANSMISSION. 1157 00:56:13,080 --> 00:56:14,800 BETWEEN HAMSTERS WITH OMICRONS 1158 00:56:14,800 --> 00:56:17,000 WE TESTED MULTIPLE OMICRON 1159 00:56:17,000 --> 00:56:20,040 VARIANTS CO-HOUSING HAMSTERS AND 1160 00:56:20,040 --> 00:56:21,240 THEY DO TRANSMIT QUITE WELL BUT 1161 00:56:21,240 --> 00:56:24,280 WE DON'T SEE THE BA 1 1162 00:56:24,280 --> 00:56:24,920 TRANSMISSION ANY MORE. 1163 00:56:24,920 --> 00:56:26,960 >> FOR EXAMPLE YOU MENTIONED THE 1164 00:56:26,960 --> 00:56:28,600 OMICRON CAN NOW INFECT SOME OF 1165 00:56:28,600 --> 00:56:30,800 THE WILD TYPE MICE, RIGHT? 1166 00:56:30,800 --> 00:56:32,280 SO THAT MEAN SOME CHANGE IN 1167 00:56:32,280 --> 00:56:36,120 USAGE OF RECEPTOR OR OTHER 1168 00:56:36,120 --> 00:56:38,680 PATHOGENICITY. 1169 00:56:38,680 --> 00:56:40,080 SO DOES THESE NEW VARIANTS WE 1170 00:56:40,080 --> 00:56:43,960 ARE NOW ENCOUNTERING, ARE THOSE 1171 00:56:43,960 --> 00:56:46,880 CHANGING IN HAMSTERS SAY TO BODY 1172 00:56:46,880 --> 00:56:48,520 TISSUES OR NEUROLOGICAL TISSUES? 1173 00:56:48,520 --> 00:56:51,200 >> YEAH THAT, IS POSSIBLE. 1174 00:56:51,200 --> 00:56:54,080 THE MUTATION IN S PROTEIN ALLOWS 1175 00:56:54,080 --> 00:56:58,440 SOME OF THE VARIANTS RECOGNIZE 1176 00:56:58,440 --> 00:57:00,120 HUMAN -- MOUSE BETTER SO THAT IS 1177 00:57:00,120 --> 00:57:02,280 WHY WE SEE REPLICATION OF 1178 00:57:02,280 --> 00:57:04,480 VARIANTS IN REGULAR MICE. 1179 00:57:04,480 --> 00:57:06,360 FOR THE SAME REASON YOU 1180 00:57:06,360 --> 00:57:09,920 DESCRIBED, IT IS POSSIBLE THAT 1181 00:57:09,920 --> 00:57:14,440 VARIANTS MAY RECOGNIZE HOST 1182 00:57:14,440 --> 00:57:15,000 MOLECULES DIFFERENT. 1183 00:57:15,000 --> 00:57:18,320 >> IN YOUR ANIMAL HAMSTERS 1184 00:57:18,320 --> 00:57:20,480 STUDIES, HOW LONG IS VIRUS 1185 00:57:20,480 --> 00:57:22,720 PERSIST IN THOSE HAMSTERS IN 1186 00:57:22,720 --> 00:57:24,880 DIFFERENT TISSUES? 1187 00:57:24,880 --> 00:57:30,800 >> USUALLY BY DAY 6, 7 VIRUS IS 1188 00:57:30,800 --> 00:57:32,240 GONE. 1189 00:57:32,240 --> 00:57:37,880 BUT WE DID ONLY INFECTIOUS VIRUS 1190 00:57:37,880 --> 00:57:41,040 TESTING SO IF WE DO EXTENSIVE 1191 00:57:41,040 --> 00:57:46,560 SEARCH BY PCR AFTER ONE MONTHS 1192 00:57:46,560 --> 00:57:48,360 WE MAY SEE SOMETHING THERE SO WE 1193 00:57:48,360 --> 00:57:49,760 SHOULD ACTUALLY DO THAT. 1194 00:57:49,760 --> 00:57:52,160 >> OKAY. 1195 00:57:52,160 --> 00:57:54,880 COMING FROM IAN. 1196 00:57:54,880 --> 00:57:55,360 IS THERE DIFFERENCIAL 1197 00:57:55,360 --> 00:57:57,040 SENSITIVITY IN OMICRON VARIANTS 1198 00:57:57,040 --> 00:57:58,760 VERSUS PROTOTYPE VARIANTS 1199 00:57:58,760 --> 00:58:00,480 VIRUSES IN YOUR SAMPLING MODELS 1200 00:58:00,480 --> 00:58:01,600 THAT YOU USED? 1201 00:58:01,600 --> 00:58:02,680 >> WE HAVEN'T DONE THAT 1202 00:58:02,680 --> 00:58:03,040 EXPERIMENT. 1203 00:58:03,040 --> 00:58:04,400 AS I MENTION I THINK WE SHOULD 1204 00:58:04,400 --> 00:58:04,800 DO THAT. 1205 00:58:04,800 --> 00:58:06,960 IT'S VERY IMPORTANT. 1206 00:58:06,960 --> 00:58:08,960 >> AND ANOTHER ONE IS DO YOU 1207 00:58:08,960 --> 00:58:11,080 KNOW DIFFERENCES IN OMICRON 1208 00:58:11,080 --> 00:58:13,080 COMPARED TO THE OTHER -- DO YOU 1209 00:58:13,080 --> 00:58:16,160 THINK UP ENDING THE SEQUENCE IN 1210 00:58:16,160 --> 00:58:18,440 VACCINE SPIKE SEQUENCE OF BA 1, 1211 00:58:18,440 --> 00:58:22,680 2, 3 OR 4 SHOULD BE PURSUED? 1212 00:58:22,680 --> 00:58:26,000 >> YES WE SHOULD, YES. 1213 00:58:26,000 --> 00:58:28,400 >> WHAT WOULD YOU RECOMMEND, 1214 00:58:28,400 --> 00:58:29,800 BECAUSE AS YOU SHOWED US WHEN 1215 00:58:29,800 --> 00:58:32,360 YOU HAVE INFECTION WITH BA 1 OR 1216 00:58:32,360 --> 00:58:33,400 2 USING DIFFERENTIAL IMMUNE 1217 00:58:33,400 --> 00:58:34,760 RESPONSE TO PREVIOUS STRAIN, 1218 00:58:34,760 --> 00:58:36,760 WHAT DO YOU THINK IS GOING ON 1219 00:58:36,760 --> 00:58:42,080 MORE LIKE ANGIOGENIC PHENOMENA 1220 00:58:42,080 --> 00:58:44,480 OR IMMUNE RESPONSE THAT YOU DO 1221 00:58:44,480 --> 00:58:46,760 NOT HAVE SAY MUCH MORE BROADER 1222 00:58:46,760 --> 00:58:47,760 PROTECTION AGAINST DIFFERENT 1223 00:58:47,760 --> 00:58:48,360 STRAINS? 1224 00:58:48,360 --> 00:58:54,080 >> WE HAVEN'T SEEN ORIGINAL 1225 00:58:54,080 --> 00:58:58,040 ANGIOGENIC PORTION YET. 1226 00:58:58,040 --> 00:59:00,280 IT REFERS TO HUMANS ANIMALS 1227 00:59:00,280 --> 00:59:02,840 DON'T RESPOND TO THE MOST RECENT 1228 00:59:02,840 --> 00:59:06,640 ONE, BUT CLEARLY MAINTAIN IMMUNE 1229 00:59:06,640 --> 00:59:07,640 RESPONSES TO OTHER VIRUS. 1230 00:59:07,640 --> 00:59:09,720 FUL SO AT LEAST WE ARE NOT 1231 00:59:09,720 --> 00:59:14,680 SEEING THIS SAME PORTION, WE SEE 1232 00:59:14,680 --> 00:59:16,960 MORE LIKE MORE BROAD CROSS 1233 00:59:16,960 --> 00:59:17,600 REACTIVITY. 1234 00:59:17,600 --> 00:59:19,320 SO AS LONG AS THAT LASTS, THAT'S 1235 00:59:19,320 --> 00:59:19,840 GOOD. 1236 00:59:19,840 --> 00:59:22,320 BUT WE WILL SEE WHETHER -- 1237 00:59:22,320 --> 00:59:25,720 BECAUSE WE ARE ONLY TWO YEARS OR 1238 00:59:25,720 --> 00:59:28,760 THREE YEARS IN PANDEMIC BEEN 1239 00:59:28,760 --> 00:59:31,960 EXPOSED TO VARIANTS. 1240 00:59:31,960 --> 00:59:34,400 SO I THINK TIME TELLS. 1241 00:59:34,400 --> 00:59:37,960 >> OUT OF THE THREE OMICRON 1242 00:59:37,960 --> 00:59:42,360 VARIANTS DIAGNOSED BA 1 BA 2 BA 1243 00:59:42,360 --> 00:59:44,480 4 BA 5 WHAT WOULD BE YOUR CHOICE 1244 00:59:44,480 --> 00:59:46,000 OF SPIKE VACCINE? 1245 00:59:46,000 --> 00:59:47,080 OFF THOSE THREE STRAINS? 1246 00:59:47,080 --> 00:59:49,320 >> THAT IS A VERY DIFFICULT 1247 00:59:49,320 --> 00:59:53,920 QUESTION BECAUSE WHETHER WE NEXT 1248 00:59:53,920 --> 00:59:58,320 WITH DESCENDENT OMICRON OR NOT. 1249 00:59:58,320 --> 01:00:01,440 SO THAT BECOMES RESISTANT IF THE 1250 01:00:01,440 --> 01:00:03,560 NEXT CIRCULATING VIRUS WILL BE 1251 01:00:03,560 --> 01:00:11,240 SEDESCENDENT OF OMICRON, THE 1252 01:00:11,240 --> 01:00:12,280 (INAUDIBLE) VACCINE CANDIDATE IN 1253 01:00:12,280 --> 01:00:15,040 THE SO CALLED BACK VERSED IMMUNE 1254 01:00:15,040 --> 01:00:17,360 RESPONSES, ANYTHING BETWEEN THE 1255 01:00:17,360 --> 01:00:19,240 ORIGINAL VIRUS AND OMICRON, 1256 01:00:19,240 --> 01:00:21,680 ANYTHING BETWEEN IN THAT MAP 1257 01:00:21,680 --> 01:00:23,600 WILL BE COVERED BY USING OMICRON 1258 01:00:23,600 --> 01:00:26,080 VARIANTS AS A VACCINE SO IN 1259 01:00:26,080 --> 01:00:26,840 BETWEEN EVERYTHING WOULD BE 1260 01:00:26,840 --> 01:00:30,800 COVERED SO OMICRON VACCINE MAY 1261 01:00:30,800 --> 01:00:33,280 BE VIABLE CHOICE BECAUSE YOU 1262 01:00:33,280 --> 01:00:33,880 RECOVER THERE. 1263 01:00:33,880 --> 01:00:36,720 >> WELL, THANK YOU SO MUCH, 1264 01:00:36,720 --> 01:00:37,280 YOSHI. 1265 01:00:37,280 --> 01:00:38,880 WONDERFUL TALK, WE HAD LOT OF 1266 01:00:38,880 --> 01:00:40,040 QUESTIONS, TIME IS SHORT SO I 1267 01:00:40,040 --> 01:00:43,280 THINK WE WILL THANK YOU, AND WE 1268 01:00:43,280 --> 01:00:44,400 LOOK FORWARD TO HEARING FROM YOU 1269 01:00:44,400 --> 01:00:44,640 AGAIN. 1270 01:00:44,640 --> 01:00:45,240 THANK YOU SO MUCH. 1271 01:00:45,240 --> 01:00:46,120 >> THANK YOU VERY MUCH. 1272 01:00:46,120 --> 01:00:56,360 APPRECIATE IT.