1 00:00:06,190 --> 00:00:07,858 >> WELL, GOOD MORNING, 2 00:00:07,858 --> 00:00:09,193 EVERYBODY, AND THANK YOU SO MUCH 3 00:00:09,193 --> 00:00:11,595 FOR JOINING US ON THIS RAINY 4 00:00:11,595 --> 00:00:13,664 TUESDAY MORNING WITH A LOT GOING 5 00:00:13,664 --> 00:00:16,067 ON ON THE CAMPUS. 6 00:00:16,067 --> 00:00:20,871 THIS SYMPOSIUM IS PART 1 OF A 7 00:00:20,871 --> 00:00:22,940 TWO-PART CELEBRATION OF THE FIVE 8 00:00:22,940 --> 00:00:24,708 MEMBERS OF THE NIH FAMILY 9 00:00:24,708 --> 00:00:28,045 ELECTED THIS YEAR TO THE 10 00:00:28,045 --> 00:00:29,747 NATIONAL ACADEMY OF THE 11 00:00:29,747 --> 00:00:30,714 SCIENCES. 12 00:00:30,714 --> 00:00:36,087 DRS. THOMAS KUNKEL, KYUNG 13 00:00:36,087 --> 00:00:38,489 KWON-CHUNG, GIORGIO TRINCHIERI, 14 00:00:38,489 --> 00:00:41,325 SANDRA WOLLEN AND STEVEN 15 00:00:41,325 --> 00:00:41,659 ROSENBERG. 16 00:00:41,659 --> 00:00:46,464 SO TODAY WE'LL HEAR FROM DRS. 17 00:00:46,464 --> 00:00:51,635 KWON-CHUNG, KUNKEL, AND 18 00:00:51,635 --> 00:00:52,136 TRINCHIERI. 19 00:00:52,136 --> 00:00:54,138 AND ON NOVEMBER 13TH, WE WILL 20 00:00:54,138 --> 00:00:56,107 HEAR FROM DRS. WOLLEN AND 21 00:00:56,107 --> 00:00:56,774 ROSENBERG. 22 00:00:56,774 --> 00:00:58,943 SO FIRST I'LL TELL YOU A LITTLE 23 00:00:58,943 --> 00:01:00,978 BIT ABOUT EACH OF TODAY'S THREE 24 00:01:00,978 --> 00:01:02,980 SPEAKERS AND THEN ALLOW THEM TO 25 00:01:02,980 --> 00:01:06,150 PROCEED WITH THEIR INSPIRATIONAL 26 00:01:06,150 --> 00:01:07,585 TALKS UNINTERRUPTED. 27 00:01:07,585 --> 00:01:09,553 HOPEFULLY WE'LL HAVE TIME FOR A 28 00:01:09,553 --> 00:01:12,523 QUESTION OR TWO AFTER EACH OF 29 00:01:12,523 --> 00:01:15,593 THE TALKS, AND PLEASE, IF YOU DO 30 00:01:15,593 --> 00:01:19,830 HAVE A QUESTION WHEN EACH TALK 31 00:01:19,830 --> 00:01:22,800 ENDS, PLEASE USE THE M THE MICRS 32 00:01:22,800 --> 00:01:24,001 ON EITHER SIDE OF THE 33 00:01:24,001 --> 00:01:28,038 AUDITORIUM. 34 00:01:28,038 --> 00:01:30,007 SO KYUNG KWON-CHUNG IS A NATIVE 35 00:01:30,007 --> 00:01:31,542 OF SOUTH KOREA, AND SHE RECEIVED 36 00:01:31,542 --> 00:01:37,681 HER UNDERGRADUATE DEGREE FROM 37 00:01:37,681 --> 00:01:38,749 ICHWA UNIVERSITY, WOMEN'S 38 00:01:38,749 --> 00:01:40,518 UNIVERSITY IN SEOUL. 39 00:01:40,518 --> 00:01:42,353 SHE RECEIVED A FULBRIGHT 40 00:01:42,353 --> 00:01:43,787 FELLOWSHIP TO PURSUE HER 41 00:01:43,787 --> 00:01:46,657 DOCTORAL STUDIES IN BACTERIOLOGY 42 00:01:46,657 --> 00:01:50,895 AT THE UNIVERSITY OF WISCONSIN. 43 00:01:50,895 --> 00:01:53,964 IN 1966, SHE JOINED NIAID AND 44 00:01:53,964 --> 00:01:58,869 THE FIELD OF MEDICAL MYCOLOGY. 45 00:01:58,869 --> 00:02:00,871 SHE HAS BEEN THE CHIEF OF THE 46 00:02:00,871 --> 00:02:02,072 MOLECULAR MICROBIOLOGY SECTION 47 00:02:02,072 --> 00:02:04,575 IN THE LABORATORY OF CLINICAL 48 00:02:04,575 --> 00:02:09,480 IMMUNOLOGY AND MICROBIOLOGY 49 00:02:09,480 --> 00:02:13,751 SINCE 1995. 50 00:02:13,751 --> 00:02:16,654 SHE RECEIVED THE LIFETIME 51 00:02:16,654 --> 00:02:17,188 ACHIEVEMENT AWARD FROM THE 52 00:02:17,188 --> 00:02:18,355 AMERICAN SOCIETY FOR 53 00:02:18,355 --> 00:02:21,525 MICROBIOLOGY IN 2017. 54 00:02:21,525 --> 00:02:23,527 AND HER TALK TODAY IS ENTITLED 55 00:02:23,527 --> 00:02:26,564 IN STEP WITH SCIENCE, A HALF 56 00:02:26,564 --> 00:02:31,135 CENTURY OF JOURNEY IN MEDICAL 57 00:02:31,135 --> 00:02:32,803 MYCOLOGY. 58 00:02:32,803 --> 00:02:35,773 DR. THOMAS KUNKEL RECEIVED HIS 59 00:02:35,773 --> 00:02:36,907 DOCTORATE IN 1977 FROM THE 60 00:02:36,907 --> 00:02:39,443 UNIVERSITY OF CINCINNATI. 61 00:02:39,443 --> 00:02:40,878 FOLLOWING A POSTDOCTORAL 62 00:02:40,878 --> 00:02:45,149 FELLOWSHIP AT THE UNIVERSITY OF 63 00:02:45,149 --> 00:02:48,852 WASHINGTON, HE JOINED NIEHS IN 64 00:02:48,852 --> 00:02:49,420 1982. 65 00:02:49,420 --> 00:02:51,288 DR. KUNKEL CURRENTLY LEADS THE 66 00:02:51,288 --> 00:02:54,225 DNA REPLICATION FIDELITY GROUP, 67 00:02:54,225 --> 00:02:57,828 WITHIN THE GENOME INTEGRITY AND 68 00:02:57,828 --> 00:02:59,463 STRUCTURAL BIOLOGY LABORATORY. 69 00:02:59,463 --> 00:03:03,300 HIS LAB STUDIES DNA REPLICATION 70 00:03:03,300 --> 00:03:06,270 AND HOW ENVIRONMENTAL STRESSES 71 00:03:06,270 --> 00:03:08,772 PERTURB THIS PROCESS. 72 00:03:08,772 --> 00:03:12,509 HIS TALK TODAY IS ENTITLED 73 00:03:12,509 --> 00:03:18,315 "ACCURACY MATTERS." 74 00:03:18,315 --> 00:03:19,717 DR. GIORGIO TRINCHIERI IS A 75 00:03:19,717 --> 00:03:21,585 NATIVE OF ITALY WHO RECEIVED HIS 76 00:03:21,585 --> 00:03:24,521 M.D. AT THE UNIVERSITY OF 77 00:03:24,521 --> 00:03:24,855 TURINO. 78 00:03:24,855 --> 00:03:29,226 HE JOINED THE NCI CENTER FOR 79 00:03:29,226 --> 00:03:31,428 CLINICAL RESEARCH IN 2006, AND 80 00:03:31,428 --> 00:03:33,731 IS CURRENTLY CHIEF OF THE 81 00:03:33,731 --> 00:03:35,466 LABORATORY OF INTEGRATIVE CANCER 82 00:03:35,466 --> 00:03:37,868 IMMUNOLOGY. 83 00:03:37,868 --> 00:03:41,905 PRIOR TO JOINING NCI, 84 00:03:41,905 --> 00:03:44,808 DR. TRINCHIERI WAS THE DIRECTOR 85 00:03:44,808 --> 00:03:47,711 OF THE LABORATORY FOR 86 00:03:47,711 --> 00:03:53,717 IMMUNOLOGICAL RESEARCH IN FRANCE 87 00:03:53,717 --> 00:03:56,086 AND AN NIH FOGARTY SCHOLAR IN 88 00:03:56,086 --> 00:03:57,655 THE LABORATORY FOR PARASITIC 89 00:03:57,655 --> 00:04:01,392 DISEASES AT NIAID. 90 00:04:01,392 --> 00:04:03,794 HIS TALK TODAY IS ENTITLED 91 00:04:03,794 --> 00:04:06,664 "IMMUNITY TO INFECTIONS, CANCER, 92 00:04:06,664 --> 00:04:09,366 AND IN BETWEEN." 93 00:04:09,366 --> 00:04:13,237 CONGRATULATIONS TO ALL OF YOU, 94 00:04:13,237 --> 00:04:16,173 AND DR. KWON-CHUNG, THE PODIUM 95 00:04:16,173 --> 00:04:17,374 IS YOURS. 96 00:04:17,374 --> 00:04:24,982 [APPLAUSE] 97 00:04:24,982 --> 00:04:25,516 >> THANK YOU. 98 00:04:25,516 --> 00:04:26,417 CAN YOU HEAR ME? 99 00:04:26,417 --> 00:04:28,686 IT'S QUITE A CHALLENGE TO TALK 100 00:04:28,686 --> 00:04:30,220 ABOUT 55 YEARS OF CAREER 101 00:04:30,220 --> 00:04:32,956 TRAJECTORY IN 30 MINUTES. 102 00:04:32,956 --> 00:04:41,465 SO I WILL START. 103 00:04:41,465 --> 00:04:43,567 AS A ROAD MAP FOR THIS TALK, WE 104 00:04:43,567 --> 00:04:44,735 INCLUDE PERSONAL BACKGROUND 105 00:04:44,735 --> 00:04:47,271 FIRST. 106 00:04:47,271 --> 00:04:50,441 AND THEN FOR THE MENTORS OF 107 00:04:50,441 --> 00:04:51,008 MYSELF. 108 00:04:51,008 --> 00:04:54,712 SINCE I SPENT -- I HAD ABOUT 70% 109 00:04:54,712 --> 00:04:58,749 OF MY WORK WAS IN CRYPTOCOCCUS, 110 00:04:58,749 --> 00:05:01,819 I'LL HAVE A BRIEF INTRODUCTION 111 00:05:01,819 --> 00:05:09,727 TO CRYPTOCOCCOSIS, AND THEN 112 00:05:09,727 --> 00:05:11,128 MILESTONE ACHIEVEMENTS AND THEIR 113 00:05:11,128 --> 00:05:12,096 IMPACTS ON THE PROGRESS OF 114 00:05:12,096 --> 00:05:13,630 MEDICAL MYCOLOGY. 115 00:05:13,630 --> 00:05:15,265 WHAT IS MILESTONE ANYWAY? 116 00:05:15,265 --> 00:05:19,203 I DEFINE THIS AS IF THAT WORK 117 00:05:19,203 --> 00:05:24,675 LAY THE FOUNDATION SO THAT THE 118 00:05:24,675 --> 00:05:26,143 PROGRESS CAN BE MADE IN NEW 119 00:05:26,143 --> 00:05:30,414 DIRECTION IN MEDICAL MYCOLOGY. 120 00:05:30,414 --> 00:05:31,849 AND THEN CONCLUSIONS AND 121 00:05:31,849 --> 00:05:34,451 ACKNOWLEDGMENT. 122 00:05:34,451 --> 00:05:42,760 FIRST BACKGROUND AND MENTORS. 123 00:05:42,760 --> 00:05:44,528 IN 1960 I APPLIED FOR GRADUATE 124 00:05:44,528 --> 00:05:45,362 STUDY IN THE U.S. BECAUSE I 125 00:05:45,362 --> 00:05:47,030 RECEIVED THE FULBRIGHT SCHOLAR 126 00:05:47,030 --> 00:05:49,967 SHIP, AND I WAS TEACHING AT 127 00:05:49,967 --> 00:05:52,469 UNIVERSITY IN SOUTH KOREA, SEOUL 128 00:05:52,469 --> 00:05:55,339 KOREA, BECAUSE THIS WAS AFTER MY 129 00:05:55,339 --> 00:05:56,974 MASTER'S DEGREE IN BACTERIOLOGY 130 00:05:56,974 --> 00:06:02,846 FROM THAT UNIVERSITY. 131 00:06:02,846 --> 00:06:04,214 I WAS ALREADY A MOTHER OF TWO 132 00:06:04,214 --> 00:06:05,883 LITTLE BOYS AT THAT TIME, AND 133 00:06:05,883 --> 00:06:10,387 THIS PERSON HERE IS NOW THAT 134 00:06:10,387 --> 00:06:13,624 TINY TODDLER, BUT NOW HE HAVE A 135 00:06:13,624 --> 00:06:19,997 P.I. AT NHLIB -- NATIONAL HEART, 136 00:06:19,997 --> 00:06:22,132 LUNG AND BLOOD INSTITUTE, AND HE 137 00:06:22,132 --> 00:06:25,335 IS A P.I. AND HE'S WORKING THREE 138 00:06:25,335 --> 00:06:30,174 FLEURS DFLOORS DOWN FROM MY OFF. 139 00:06:30,174 --> 00:06:33,911 MY PH.D. WORK STARTED AT THE 140 00:06:33,911 --> 00:06:35,179 UNIVERSITY OF WISCONSIN IN 1961. 141 00:06:35,179 --> 00:06:38,916 IF YOU WANT TO FIGURE OUT WHAT 142 00:06:38,916 --> 00:06:41,552 IS -- WHAT DOES 1961 LOOK LIKE? 143 00:06:41,552 --> 00:06:45,923 THIS IS EIGHT YEARS AFTER DNA 144 00:06:45,923 --> 00:06:47,791 STRUCTURE WAS FOUND OR AT LEAST 145 00:06:47,791 --> 00:06:53,263 BECAME CLEAR. 146 00:06:53,263 --> 00:06:55,265 SO WHAT'S DNA STRUCTURE AND 147 00:06:55,265 --> 00:06:58,469 EIGHT YEARS AFTER THAT, AND 10 148 00:06:58,469 --> 00:07:03,974 YEARS BEFORE, THE FIRST 149 00:07:03,974 --> 00:07:06,610 RESTRICTION ENDONUCLEASE BECAME 150 00:07:06,610 --> 00:07:07,678 AVAILABLE. 151 00:07:07,678 --> 00:07:08,779 SO FIRST YEAR GRADUATE SCHOOL 152 00:07:08,779 --> 00:07:10,614 WAS SUPPORTED BY THE FULBRIGHT, 153 00:07:10,614 --> 00:07:12,816 AS I SAID, AND FELLOWSHIP WAS 154 00:07:12,816 --> 00:07:13,684 MANAGED BY U.S. STATE 155 00:07:13,684 --> 00:07:15,118 DEPARTMENT. 156 00:07:15,118 --> 00:07:19,723 AND THE REMAINING YEARS, THE NSF 157 00:07:19,723 --> 00:07:23,227 RESEARCH GRANT FOR THREE YEARS, 158 00:07:23,227 --> 00:07:27,364 BECAUSE DR. RAPER HAD AN NSF 159 00:07:27,364 --> 00:07:30,701 GRANT TO MAKE A NEW MONO GRAPH. 160 00:07:30,701 --> 00:07:32,569 THEN I COMPLETED DOCTORATE IN 161 00:07:32,569 --> 00:07:32,970 1965. 162 00:07:32,970 --> 00:07:35,472 AND I'M VERY SORRY FOR THIS 163 00:07:35,472 --> 00:07:38,108 FUZZY PICTURE, BUT DR. RAPER AND 164 00:07:38,108 --> 00:07:41,578 I WERE STANDING IN FRONT OF THE 165 00:07:41,578 --> 00:07:44,448 BUILDING OF BACTERIOLOGY IN 166 00:07:44,448 --> 00:07:48,385 UNIVERSITY OF WISCONSIN IN '61. 167 00:07:48,385 --> 00:07:50,454 WHO IS PROFESSOR KENNETH RAPER 168 00:07:50,454 --> 00:07:50,687 ANYWAY? 169 00:07:50,687 --> 00:07:52,856 MOST OF YOU MAY HAVE NOT HEARD 170 00:07:52,856 --> 00:07:55,292 ABOUT HIM. 171 00:07:55,292 --> 00:08:02,599 HE IS FATHER OF CYST AT SYSTEMN 172 00:08:02,599 --> 00:08:05,102 THE MOST COMMON MOLD LIKE 173 00:08:05,102 --> 00:08:11,341 ASPERGILLUS, PENICILLUM AND 174 00:08:11,341 --> 00:08:13,076 CELLULAR SLIME MOLD. 175 00:08:13,076 --> 00:08:16,480 HE DISCOVERED VERY HIGH 176 00:08:16,480 --> 00:08:21,618 PENICILLIN YIELDING STRAIN OF 177 00:08:21,618 --> 00:08:25,022 PENICILLUM CRISOJENUM AND PFIZER 178 00:08:25,022 --> 00:08:27,391 USED THIS STRAIN AND AS A 179 00:08:27,391 --> 00:08:30,394 RESULT, WHOLESALE COST OF 10,000 180 00:08:30,394 --> 00:08:36,266 UNITS OF PENICILLIN WAS $20 181 00:08:36,266 --> 00:08:38,101 THEN, BECAME 3 CENTS BY WORLD 182 00:08:38,101 --> 00:08:42,306 WAR II BECAUSE OF HIS STRAIN. 183 00:08:42,306 --> 00:08:44,908 HE DISCOVERED AND DESCRIBED THE 184 00:08:44,908 --> 00:08:50,948 LIFECYCLE OF CELLULAR SLIME MOLD 185 00:08:50,948 --> 00:08:53,216 IN 1933. 186 00:08:53,216 --> 00:08:54,918 WHICH BECAME MODERN ORGANISM FOR 187 00:08:54,918 --> 00:09:01,892 CELL BIOLOGY RESEARCH. 188 00:09:01,892 --> 00:09:04,294 MY CAREER IN MYCOLOGY TOOK OFF 189 00:09:04,294 --> 00:09:11,735 AFTER THE D THE DISCOVERY WHICHI 190 00:09:11,735 --> 00:09:12,936 DISCOVERED WHEN I WAS IN 191 00:09:12,936 --> 00:09:14,371 GRADUATE SCHOOL BECAUSE 192 00:09:14,371 --> 00:09:16,640 DR. RAPER GAVE ME THE RESEARCH 193 00:09:16,640 --> 00:09:20,377 PROJECT SAYING ISOLATE AS MANY 194 00:09:20,377 --> 00:09:23,981 NEW SPECIES OF ASPERGILLUS AND 195 00:09:23,981 --> 00:09:26,550 DESCRIBE PHENOTYPICALLY, 196 00:09:26,550 --> 00:09:26,917 BIOCHEMICALLY. 197 00:09:26,917 --> 00:09:29,786 SO THAT WAS MY PROJECT, AND 198 00:09:29,786 --> 00:09:31,989 WHERE WAS THIS ASPERGILLUS 199 00:09:31,989 --> 00:09:32,489 COMING FROM? 200 00:09:32,489 --> 00:09:35,392 IT WAS COMING FROM SOIL 201 00:09:35,392 --> 00:09:37,427 COLLECTED IN COSTA RICA. 202 00:09:37,427 --> 00:09:40,063 WHY COSTA RICA? 203 00:09:40,063 --> 00:09:42,566 BECAUSE MOST OF THE ASPERGILLUS 204 00:09:42,566 --> 00:09:48,372 AND PENECILLUM WERE FROM TREM 205 00:09:48,372 --> 00:09:49,806 PRATT ZONE CLIMATE AND VERY 206 00:09:49,806 --> 00:09:52,643 LITTLE WAS KNOWN ABOUT THOSE 207 00:09:52,643 --> 00:09:54,177 ASPERGILLUS POPULATION IN 208 00:09:54,177 --> 00:09:58,882 TROPICAL SOIL. 209 00:09:58,882 --> 00:10:02,152 BECAUSE HE GOT NSF GRANT, HE HAD 210 00:10:02,152 --> 00:10:05,889 TO INCLUDE AS MANY SPECIES AS HE 211 00:10:05,889 --> 00:10:08,291 COULD. 212 00:10:08,291 --> 00:10:10,360 AND I DISCOVERED NINE NEW 213 00:10:10,360 --> 00:10:11,895 SPECIES DISCOVERED FROM COSTA 214 00:10:11,895 --> 00:10:13,430 RICA SOIL AND THEN ONE OF THEM 215 00:10:13,430 --> 00:10:18,568 TURNED OUT TO BE THE FIRST 216 00:10:18,568 --> 00:10:21,471 HETEROTHALLIC SPECIES IN THE 217 00:10:21,471 --> 00:10:23,173 GENUS ASPERGILLUS WHICH IS 218 00:10:23,173 --> 00:10:26,677 COMPOSED OF 132 SPECIES. 219 00:10:26,677 --> 00:10:28,512 NOW, WHAT IS THE HETEROTHAT 220 00:10:28,512 --> 00:10:29,079 WILLISM? 221 00:10:29,079 --> 00:10:40,657 FIRST TIME IS HOMOTHALLISMTHAT E 222 00:10:46,329 --> 00:10:48,265 SEXUAL STATE IN PURE CULTURE. 223 00:10:48,265 --> 00:10:53,236 IN OTHER WORDS, IT'S A 224 00:10:53,236 --> 00:10:53,804 HERMAPHRODITE. 225 00:10:53,804 --> 00:10:57,941 SECOND TYPE IS CALLED 226 00:10:57,941 --> 00:10:59,376 HETEROTHALLISM. 227 00:10:59,376 --> 00:11:00,777 IT'S SELF STERILE BUT CROSS 228 00:11:00,777 --> 00:11:01,545 FERTILE. 229 00:11:01,545 --> 00:11:03,980 NOW, IT REQUIRES MATING WITH 230 00:11:03,980 --> 00:11:08,685 OPPOSITE MATING TYPE. 231 00:11:08,685 --> 00:11:10,420 AND TO COMPLETE SEXUAL 232 00:11:10,420 --> 00:11:10,687 LIFECYCLE. 233 00:11:10,687 --> 00:11:13,890 SO I FOUND THIS HETEROTHALLIC AS 234 00:11:13,890 --> 00:11:17,327 PASPERGILLUS FROM COSTA RECAN 235 00:11:17,327 --> 00:11:19,963 SOIL AND THAT REALLY BECAME THE 236 00:11:19,963 --> 00:11:23,233 LAUNCHING PAD FOR MY MYCOLOGICAL 237 00:11:23,233 --> 00:11:23,467 CAREER. 238 00:11:23,467 --> 00:11:26,503 WHERE DID I GET MEDICAL MYCOLOGY 239 00:11:26,503 --> 00:11:30,440 TRAINING AT NIH? 240 00:11:30,440 --> 00:11:32,642 DR. CHESTER EMMONS, WHO USED TO 241 00:11:32,642 --> 00:11:35,178 BE THE SECTION CHIEF OF MEDICAL 242 00:11:35,178 --> 00:11:41,485 MYCOLOGY AT THE NIAID, HE IS A 243 00:11:41,485 --> 00:11:43,019 LEGENDARY PIONEER IN PATHOGENIC 244 00:11:43,019 --> 00:11:44,454 MYCOLOGY IN THE UNITED STATES. 245 00:11:44,454 --> 00:11:48,058 AND HE HIRED ME AS A VISITING 246 00:11:48,058 --> 00:11:49,860 FELLOW IN MEDICAL MYCOLOGY 247 00:11:49,860 --> 00:11:52,662 SECTION. 248 00:11:52,662 --> 00:11:55,732 AND HE DISCOVERED THE ECOLOGICAL 249 00:11:55,732 --> 00:11:59,102 NATURE OF ALMOST EVERY IMPORTANT 250 00:11:59,102 --> 00:12:02,272 SYSTEMIC FUNGAL DISEASE SUCH AS 251 00:12:02,272 --> 00:12:05,909 HISTOPLASMOSIS, CRYPT 252 00:12:05,909 --> 00:12:07,377 TOECOCCOSIS AND HE'S THE ONE 253 00:12:07,377 --> 00:12:12,682 THAT FOUND THE ASSOCIATION OF 254 00:12:12,682 --> 00:12:14,217 COCCIDIOIDES WITH DESERT 255 00:12:14,217 --> 00:12:17,721 RODENTS. 256 00:12:17,721 --> 00:12:19,556 HIS THREE EDITIONS OF MEDICAL 257 00:12:19,556 --> 00:12:20,757 MYCOLOGY SERVED AS THE REFERENCE 258 00:12:20,757 --> 00:12:22,225 FOR MEDICAL MYCOLOGY IN GRADUATE 259 00:12:22,225 --> 00:12:23,660 SCHOOL AND MEDICAL SCHOOL FOR 260 00:12:23,660 --> 00:12:27,564 MORE THAN THREE DECADES. 261 00:12:27,564 --> 00:12:29,833 TO MY DISAPPOINTMENT, HE 262 00:12:29,833 --> 00:12:34,337 ANNOUNCED RETIREMENT AND GONE 263 00:12:34,337 --> 00:12:35,438 AFTER SIX MONTHS AFTER MY 264 00:12:35,438 --> 00:12:40,577 ARRIVAL, AND SO I WAS FORCED TO 265 00:12:40,577 --> 00:12:43,513 FUNCTION AS AN INVESTIGATOR ON 266 00:12:43,513 --> 00:12:47,117 MY OWN AND DIDN'T REALLY GET THE 267 00:12:47,117 --> 00:12:49,419 BENEFIT OF BEING TRAINED BY 268 00:12:49,419 --> 00:12:53,590 DR. EMMONS. 269 00:12:53,590 --> 00:12:56,760 IN 1974, I JOINED CLINICAL 270 00:12:56,760 --> 00:12:59,262 MYCOLOGY SECTION HEADED BY JOHN 271 00:12:59,262 --> 00:13:01,998 BENNETT IN THE LEVEL THREE OF 272 00:13:01,998 --> 00:13:02,499 CLINICAL INVESTIGATION. 273 00:13:02,499 --> 00:13:04,835 AS YOU CAN SEE HERE, WHEN IT 274 00:13:04,835 --> 00:13:08,338 COMES TO DIVERSITY, ZERO. 275 00:13:08,338 --> 00:13:09,539 [LAUGHTER] 276 00:13:09,539 --> 00:13:12,742 ZERO DIVERSITY IN RACE, GENDER, 277 00:13:12,742 --> 00:13:14,344 AND ACADEMIC DEGREE. 278 00:13:14,344 --> 00:13:17,047 EVERYONE WAS M.D., EVERYONE WAS 279 00:13:17,047 --> 00:13:19,616 WHITE MALE, AND THEN I POINTED 280 00:13:19,616 --> 00:13:21,251 MYSELF WITH YELLOW ARROW. 281 00:13:21,251 --> 00:13:25,088 CAN YOU SEE IT? 282 00:13:25,088 --> 00:13:32,395 SO MY PRESENCE MADE TO LCI AS 283 00:13:32,395 --> 00:13:34,931 98% HOMOLOGY INSTEAD OF 100%. 284 00:13:34,931 --> 00:13:37,200 WELL, IT WASN'T A VERY 285 00:13:37,200 --> 00:13:38,101 COMFORTABLE ENVIRONMENT, BECAUSE 286 00:13:38,101 --> 00:13:43,340 I FELT LIKE AN ODD BALL. 287 00:13:43,340 --> 00:13:47,844 BUT THE SAVIOR WAS JOHN BENNETT. 288 00:13:47,844 --> 00:13:52,315 HE GAVE ME THE CHANCE TO BUILD A 289 00:13:52,315 --> 00:13:58,889 PROJECT PROGRAM ON CRYPTO COCCIS 290 00:13:58,889 --> 00:14:01,057 AND CRYPTO COCCOSIS. 291 00:14:01,057 --> 00:14:02,692 WHEN I JOINED, HE JUST OPENED 292 00:14:02,692 --> 00:14:09,032 HIS RESOURCES FOR ME TO USE. 293 00:14:09,032 --> 00:14:11,434 HE HAD NEARLY 1,000 STRAINS OF 294 00:14:11,434 --> 00:14:19,943 ZESEROTYPED CRYPTOFORMENS IN HIS 295 00:14:19,943 --> 00:14:20,810 STOCK AND HE HAD ALL THE 296 00:14:20,810 --> 00:14:21,544 INFORMATION ABOUT WHERE THIS IS 297 00:14:21,544 --> 00:14:23,079 COMING FROM, WHETHER IT'S A 298 00:14:23,079 --> 00:14:24,748 CLINICAL SOURCE OR ENVIRONMENT 299 00:14:24,748 --> 00:14:27,817 SOURCE, IF IT IS, WHAT KIND OF A 300 00:14:27,817 --> 00:14:30,687 SAMPLE OR CLINICAL SAMPLE DOES 301 00:14:30,687 --> 00:14:35,692 THE ISOLATE COME FROM. 302 00:14:35,692 --> 00:14:39,296 SO SINCE I WORKED 70% OF MY 303 00:14:39,296 --> 00:14:41,598 CAREER ON CRYPTO COCCUS, I'D 304 00:14:41,598 --> 00:14:43,233 LIKE TO INTRODUCE BRIEFLY ABOUT 305 00:14:43,233 --> 00:14:51,007 THE FUNGUS AND THE DISEASE. 306 00:14:51,007 --> 00:14:52,409 AS YOU CAN SEE HERE, THE DISEASE 307 00:14:52,409 --> 00:14:54,077 STARTS FROM INHALATION, AND IT 308 00:14:54,077 --> 00:14:59,749 WILL MULTIPLY IN THE LUNG IF THE 309 00:14:59,749 --> 00:15:03,486 INDIVIDUAL HAS SOME IMMUNE 310 00:15:03,486 --> 00:15:04,120 DEFECT ISSUE. 311 00:15:04,120 --> 00:15:11,027 AND THEN IT WILL SPREAD AND 312 00:15:11,027 --> 00:15:13,096 CROSS THE BLOOD BRAIN BARRIER 313 00:15:13,096 --> 00:15:18,335 AND CAUSE FULL FULMINATING BRAN 314 00:15:18,335 --> 00:15:21,304 DISEASE, WHICH IS 100% FATAL 315 00:15:21,304 --> 00:15:25,241 UNLESS TREATED. 316 00:15:25,241 --> 00:15:30,714 AND THE RISK FACTORS, 317 00:15:30,714 --> 00:15:31,848 HIV-POSITIVE AIDS PATIENTS AT 318 00:15:31,848 --> 00:15:34,718 THE HIGHEST RISK. 319 00:15:34,718 --> 00:15:42,425 IDIOPATHIC CD4 CYTOPENIA AND 320 00:15:42,425 --> 00:15:46,396 CORTICOSTEROID TREATMENT, OR 321 00:15:46,396 --> 00:15:47,731 ORGAN TRANSPLANTATION OR EVEN 322 00:15:47,731 --> 00:15:50,100 CANCER, THESE ARE THE RISK 323 00:15:50,100 --> 00:15:56,539 FACTORS FOR CRYPTOCOCCISIS. 324 00:15:56,539 --> 00:15:59,943 AND ITS MOST COMMON CAUSE OF 325 00:15:59,943 --> 00:16:05,515 DEATH IS MENINGOENCEPHALITIS. 326 00:16:05,515 --> 00:16:07,817 ACTUALLY THIS IS THE MOST COMMON 327 00:16:07,817 --> 00:16:12,756 CAUSE OF MENINGITIS, NOT VIRAL, 328 00:16:12,756 --> 00:16:17,327 IF YOU REMOVE THE -- THIS IS THE 329 00:16:17,327 --> 00:16:18,428 MOST COMMON MENINGITIS CAUSE IN 330 00:16:18,428 --> 00:16:27,137 THE U.S. 331 00:16:27,137 --> 00:16:28,338 HERE I'M SHOWING VERY 332 00:16:28,338 --> 00:16:29,039 INTERESTING LOOKING BRAIN, OF 333 00:16:29,039 --> 00:16:29,439 COURSE. 334 00:16:29,439 --> 00:16:30,507 THE LEFT, YOU CAN SEE THE BRAIN 335 00:16:30,507 --> 00:16:32,942 IS COVERED WITH A WHITE. 336 00:16:32,942 --> 00:16:41,518 THIS MEANS A CRYPTOCOCCIS IS 337 00:16:41,518 --> 00:16:42,886 GROWING VERY WELL AND THE OTHER 338 00:16:42,886 --> 00:16:50,226 ONE IS A BRAIN SECTION, AND THIS 339 00:16:50,226 --> 00:16:52,862 GLISTENING AREA MEANS -- AS WELL 340 00:16:52,862 --> 00:16:54,397 AS FUNGAL GROWTH. 341 00:16:54,397 --> 00:16:59,969 NOW, THIS FUNGUS IS NEUROTROPIC 342 00:16:59,969 --> 00:17:02,038 BECAUSE IT HAS THE ABILITY TO 343 00:17:02,038 --> 00:17:05,108 CROSS THE BLOOD BRAIN BARRIER 344 00:17:05,108 --> 00:17:07,844 FIRST, AND THEN IT CAN SURVIVE 345 00:17:07,844 --> 00:17:11,748 IN VERY LOW GLUCOSE ENVIRONMENT. 346 00:17:11,748 --> 00:17:18,755 AND THEN IT HAS ENZYME CALLED 347 00:17:18,755 --> 00:17:25,995 LACCASE, WHICH OXIDIZES DOPAM 348 00:17:25,995 --> 00:17:28,298 DOPAMINE -- AND NOREPINEPHRINE, 349 00:17:28,298 --> 00:17:28,832 NEUROTRANSMITTERS. 350 00:17:28,832 --> 00:17:32,669 NOW I'M GOING TO SHOW CHRONOLOGY 351 00:17:32,669 --> 00:17:33,770 OF MILESTONE RESEARCH. 352 00:17:33,770 --> 00:17:36,806 AS I SAID, MILESTONE IF IT LAY 353 00:17:36,806 --> 00:17:37,474 THE FOUNDATION FOR A NEW 354 00:17:37,474 --> 00:17:43,580 DIRECTION OF A STUDY IN 355 00:17:43,580 --> 00:17:45,048 CRYPTCOCCUS AND CRYPTOCOCCOSIS. 356 00:17:45,048 --> 00:17:46,649 IF YOU LOOK AT THIS SLIDE, THE 357 00:17:46,649 --> 00:17:50,487 LOWER PART IS THE DEVELOPMENT 358 00:17:50,487 --> 00:17:51,354 OF -- TECHNOLOGICAL DEVELOPMENT 359 00:17:51,354 --> 00:17:53,223 WHICH IS RELEVANT FOR OUR STUDY, 360 00:17:53,223 --> 00:17:58,695 AND THEN I HIGHLIGHTED THE PARTS 361 00:17:58,695 --> 00:18:02,198 WHICH ARE ALL CRYPTOCOCCAL 362 00:18:02,198 --> 00:18:03,266 RESEARCH. 363 00:18:03,266 --> 00:18:10,640 AND THE MOST IMPORTANT PART IS I 364 00:18:10,640 --> 00:18:14,077 DISCOVERED HETEROTHALLISM IN TWO 365 00:18:14,077 --> 00:18:16,713 AGENTS OF CRYPTOCOCCOSIS IN 1975 366 00:18:16,713 --> 00:18:17,147 AND 1976. 367 00:18:17,147 --> 00:18:19,883 SO THIS IS A YEAR AFTER I 368 00:18:19,883 --> 00:18:24,487 ARRIVED AT THE BENNETT 369 00:18:24,487 --> 00:18:28,424 LABORATORY, I ASKED HIM, CAN YOU 370 00:18:28,424 --> 00:18:30,493 COLLECT -- JUST PICK 20 ISOLATES 371 00:18:30,493 --> 00:18:32,262 FROM HALF CLINICAL AND HALF 372 00:18:32,262 --> 00:18:37,167 ENVIRONMENTAL FROM YOUR 1,000 373 00:18:37,167 --> 00:18:39,536 STOCK CULTURES, AND HE PICKED 374 00:18:39,536 --> 00:18:41,604 20, AND THEN I WAS THINKING 375 00:18:41,604 --> 00:18:45,475 MAYBE IT'S A HETEROTHALLISM AND 376 00:18:45,475 --> 00:18:52,482 STUDIED FROM THERE. 377 00:18:52,482 --> 00:18:56,319 I FORMULATED CGB MEDIA THAT CAN 378 00:18:56,319 --> 00:19:00,023 SEPARATE THE TWO CRYPT TOECOCCAL 379 00:19:00,023 --> 00:19:00,924 PATHOGENS, BECAUSE AT THAT TIME 380 00:19:00,924 --> 00:19:04,194 YOU REMEMBER THERE WAS NO DNA 381 00:19:04,194 --> 00:19:07,697 SEQUENCE OR -- AND NO MOLOTOV 382 00:19:07,697 --> 00:19:08,464 FOR SURE. 383 00:19:08,464 --> 00:19:12,035 SO FOR THE IDENTIFICATION OF 384 00:19:12,035 --> 00:19:14,671 THESE TWO SPECIES WAS QUITE 385 00:19:14,671 --> 00:19:18,074 LABORIOUS, BECAUSE YOU HAD TO 386 00:19:18,074 --> 00:19:20,376 HAVE EITHER ANTIBODIES SPECIFIC 387 00:19:20,376 --> 00:19:24,080 TO EACH SEROTYPE, AND YOU KNOW 388 00:19:24,080 --> 00:19:26,082 HOW IT COST AND THE TIME THAT'S 389 00:19:26,082 --> 00:19:31,754 SPENT TO GET THE ANTIBODIES. 390 00:19:31,754 --> 00:19:34,490 SO THIS MEDIA REALLY LAID THE 391 00:19:34,490 --> 00:19:36,226 FOUNDATION FOR LOTS OF THINGS. 392 00:19:36,226 --> 00:19:40,396 AND THEN DISCOVERED MOST 393 00:19:40,396 --> 00:19:42,899 IMPORTANT BASIC VIRULENCE 394 00:19:42,899 --> 00:19:49,272 FACTORS OF CRYPT COCCUS IN '82. 395 00:19:49,272 --> 00:19:54,277 AND THEN TRANSFORMATION OF 396 00:19:54,277 --> 00:19:55,178 CRYPTOCOCCUS, DNA MEDIATED 397 00:19:55,178 --> 00:19:57,614 TRANSFORMATION IN 1990, AND THEN 398 00:19:57,614 --> 00:20:02,452 CREATED A CONGENIC SET AT ONE OF 399 00:20:02,452 --> 00:20:03,953 THE STRAIN THAT I CREATED WAS 400 00:20:03,953 --> 00:20:06,456 USED FOR THE FIRST GENOME 401 00:20:06,456 --> 00:20:08,858 SEQUENCING STRAIN OF 402 00:20:08,858 --> 00:20:11,961 CRYPTOCOCCUS. 403 00:20:11,961 --> 00:20:15,632 AND ONCE FRANCE FORMATION 404 00:20:15,632 --> 00:20:16,799 TECHNIQUE WAS ACHIEVED AND FIRST 405 00:20:16,799 --> 00:20:20,136 THING WE DID WAS MOLECULAR 406 00:20:20,136 --> 00:20:23,106 KOCH'S POSTULATE WAS FULL FIELD 407 00:20:23,106 --> 00:20:24,607 IN 1994. 408 00:20:24,607 --> 00:20:28,244 AND WE IDENTIFIED THE FIRST 409 00:20:28,244 --> 00:20:32,181 FUNGAL QUORUM SENSES PEPTIDE 410 00:20:32,181 --> 00:20:34,684 BECAUSE THEY WERE ALL ORGANIC 411 00:20:34,684 --> 00:20:37,754 ASSAY AND PEPTIDE WAS ONLY FOUND 412 00:20:37,754 --> 00:20:39,922 IN -- BUT WE FOUND THE 413 00:20:39,922 --> 00:20:42,992 CRYPTOCOCCUS HAS THE FIRST 414 00:20:42,992 --> 00:20:46,596 PEPTIDE QUORUM SENSING MATERIAL. 415 00:20:46,596 --> 00:20:50,767 AND THEN AZOLE HETERORESISTANCE 416 00:20:50,767 --> 00:20:54,804 IN GM-CSF AUTOANTIBODIES AS A 417 00:20:54,804 --> 00:20:59,542 MAJOR RISK FACTOR FOR C. GATTII, 418 00:20:59,542 --> 00:21:01,144 BUT I CANNOT GO THROUGH ALL THIS 419 00:21:01,144 --> 00:21:02,679 AND I'M JUST GOING TO 420 00:21:02,679 --> 00:21:04,647 CONCENTRATE ON THE FIRST FIVE 421 00:21:04,647 --> 00:21:08,251 SUBJECTS, BECAUSE THAT'S REALLY 422 00:21:08,251 --> 00:21:10,787 THE MAJOR FOUNDATION WE LAID FOR 423 00:21:10,787 --> 00:21:13,623 THE PROGRESS. 424 00:21:13,623 --> 00:21:18,761 SO HOW DID THIS WORK WHICH I 425 00:21:18,761 --> 00:21:22,265 CALL MILESTONE WORK LAY THE 426 00:21:22,265 --> 00:21:25,468 FOUNDATION FOR ADVANCEMENT OF 427 00:21:25,468 --> 00:21:30,340 WHAT FIELD OF RESEARCH? 428 00:21:30,340 --> 00:21:34,077 FIRST CRYPTOCOCCUS WAS NOT EVEN 429 00:21:34,077 --> 00:21:36,245 CLASSIFIED PROPERLY BECAUSE IT 430 00:21:36,245 --> 00:21:38,114 DIDN'T HAVE A SEXUAL STATE 431 00:21:38,114 --> 00:21:39,515 BEFORE. 432 00:21:39,515 --> 00:21:46,322 AND THEN PHYLOGENY OF THE FUNGAL 433 00:21:46,322 --> 00:21:48,358 KINGDOM COULD BE STUDIED 434 00:21:48,358 --> 00:21:50,360 INCLUDING CRYPTOCOCCUS, AND 435 00:21:50,360 --> 00:21:51,227 PATHOBIOLOGY FIELD 436 00:21:51,227 --> 00:21:52,995 IDENTIFICATION OF ESSENTIAL 437 00:21:52,995 --> 00:21:56,432 VIRULENCE FACTORS, AND THE 438 00:21:56,432 --> 00:22:00,002 FULFILLMENT OF MOLECULAR COST 439 00:22:00,002 --> 00:22:01,637 POSTULATE FOR VIRULENCE FACTOR, 440 00:22:01,637 --> 00:22:05,274 AND THEN ECOLOGY IDENTIFIED THE 441 00:22:05,274 --> 00:22:11,147 NEW SOURCE OF ECOLOGICAL NICHE 442 00:22:11,147 --> 00:22:12,448 OF CRYPTO COCCI. 443 00:22:12,448 --> 00:22:15,618 WHEN IT COMES TO EPIDEMIOLOGY, 444 00:22:15,618 --> 00:22:21,224 EPIDEMIOLOGY OF THE TWO SPECIES 445 00:22:21,224 --> 00:22:23,826 WHICH CAUSE CRYPTOCOCCOSIS WAS 446 00:22:23,826 --> 00:22:32,969 QUITE DIFFERENT. 447 00:22:32,969 --> 00:22:40,943 IN GENETICS -- AND IN CELL 448 00:22:40,943 --> 00:22:42,044 BIOLOGY, IDENTIFICATION OF THE 449 00:22:42,044 --> 00:22:44,547 FIRST FUNGAL DENSITY-DEPENDENT 450 00:22:44,547 --> 00:22:49,585 QUORUM SENSING PEPTIDE AND THE 451 00:22:49,585 --> 00:22:52,422 HETERORESISTANCE TO AZOLE DRUGS. 452 00:22:52,422 --> 00:22:54,791 SO SINCE I CANNOT COVER ALL 453 00:22:54,791 --> 00:22:57,660 THIS, I'M GOING TO JUST TALK 454 00:22:57,660 --> 00:23:00,296 ABOUT THIS IN DETAIL, AS I SAID 455 00:23:00,296 --> 00:23:04,367 BECAUSE THIS WAS REALLY THE VERY 456 00:23:04,367 --> 00:23:05,368 BASIC FOUNDATION OF KNOWLEDGE 457 00:23:05,368 --> 00:23:07,203 THAT WAS NEEDED FOR 458 00:23:07,203 --> 00:23:13,209 CRYPTOCOCCUS. 459 00:23:13,209 --> 00:23:16,179 NOW WHEN I JOINED DR. BENNETT'S 460 00:23:16,179 --> 00:23:20,082 OFFICE/LABORATORY IN '74, I WAS 461 00:23:20,082 --> 00:23:23,386 GIVEN A CHANCE TO STUDY KR CRYP, 462 00:23:23,386 --> 00:23:26,122 BUT I HAD TO HAVE A GOAL FIRST. 463 00:23:26,122 --> 00:23:29,292 SO MY FIRST GOAL WAS UNCOVERING 464 00:23:29,292 --> 00:23:33,329 THE COMPLETE LIFECYCLE OF 465 00:23:33,329 --> 00:23:34,430 CRYPTOCOCCUS BY INVESTIGATING 466 00:23:34,430 --> 00:23:35,631 THE POSSIBILITY OF 467 00:23:35,631 --> 00:23:36,199 HETEROTHALLISM BECAUSE UNTIL 468 00:23:36,199 --> 00:23:39,902 THIS TIME, CRYPTOCOCCUS WAS 469 00:23:39,902 --> 00:23:41,437 KNOWN AS A BUDDING YEAST, THAT'S 470 00:23:41,437 --> 00:23:44,941 ALL. 471 00:23:44,941 --> 00:23:46,776 AND DISCOVERED THE 472 00:23:46,776 --> 00:23:51,280 HETEROTHALLISM LIFECYCLE IN 473 00:23:51,280 --> 00:23:52,915 NEOFORMENS IN 1975 AND THE OTHER 474 00:23:52,915 --> 00:23:56,519 SPECIES IN 1976, AND RESOLVE THE 475 00:23:56,519 --> 00:23:59,789 TAXONOMY POSITION OF 476 00:23:59,789 --> 00:24:01,123 CRYPTOCOCCUS WITHIN -- AND 477 00:24:01,123 --> 00:24:03,292 ESTABLISHED THE FOUNDATION FOR 478 00:24:03,292 --> 00:24:08,898 THE PHYLOGENETIC STUDY. 479 00:24:08,898 --> 00:24:13,903 NOW, THIS IS THE WAY THE FUNGUS 480 00:24:13,903 --> 00:24:15,805 WAS KNOWN FROM THE TIME OF 481 00:24:15,805 --> 00:24:20,309 DISCOVERY WAS 1894 UNTIL 1975, 482 00:24:20,309 --> 00:24:22,645 WHEN I DISCOVERED THE SEXUAL 483 00:24:22,645 --> 00:24:22,979 STATE. 484 00:24:22,979 --> 00:24:25,081 NOW THIS IS JUST A SCANNING 485 00:24:25,081 --> 00:24:29,452 ELECTRON MICROSCOPY SHOWING THAT 486 00:24:29,452 --> 00:24:32,388 IT'S COVERED WITH POLICY 487 00:24:32,388 --> 00:24:35,224 SACCHARIDE CAPSULE. 488 00:24:35,224 --> 00:24:37,760 WHEN I FOUND THE LIFECYCLE, I 489 00:24:37,760 --> 00:24:42,665 WAS NOT ONLY HAPPY BUT WHOLE 490 00:24:42,665 --> 00:24:43,966 THINGS START OPENING UP. 491 00:24:43,966 --> 00:24:47,937 FIRST, LIFECYCLE HERE, THIS 492 00:24:47,937 --> 00:24:52,975 IS -- I CALL MATTA AND MATT 493 00:24:52,975 --> 00:24:54,243 ALPHA BECAUSE THERE WERE TWO 494 00:24:54,243 --> 00:24:55,378 SEX, ALPHA AND A. 495 00:24:55,378 --> 00:24:57,446 I JUST NAMED IT AS ALPHA AND A. 496 00:24:57,446 --> 00:24:59,749 THERE'S NO REASON WHY IT CANNOT 497 00:24:59,749 --> 00:25:01,050 BE PLUS OR MINUS OR SOMETHING 498 00:25:01,050 --> 00:25:01,384 ELSE. 499 00:25:01,384 --> 00:25:06,856 AND WHEN YOU MIX THESE YEAST 500 00:25:06,856 --> 00:25:08,925 CELLS, ON THE MEDIA, WHICH IS 501 00:25:08,925 --> 00:25:12,094 CONDUCIVE FOR THE SEXUAL 502 00:25:12,094 --> 00:25:15,498 REPRODUCTION, THEN THIS -- YOU 503 00:25:15,498 --> 00:25:18,434 CAN SEE THE ZONE HERE COMPLETELY 504 00:25:18,434 --> 00:25:20,069 DIFFERENT MORPHOLOGY, AND THIS 505 00:25:20,069 --> 00:25:22,038 IS WHAT IT WAS. 506 00:25:22,038 --> 00:25:25,441 FIRST MATING TYPE A AND ALPHA 507 00:25:25,441 --> 00:25:28,177 JUST BUD AND BUD, BUT WHEN THEY 508 00:25:28,177 --> 00:25:35,851 MEET TOGETHER, THEY CON JEW GATE 509 00:25:35,851 --> 00:25:45,895 AND PRODUCE THIS FLUFFY WHITE 510 00:25:45,895 --> 00:25:46,329 AREA. 511 00:25:46,329 --> 00:25:49,465 AND THE NUCLEAR FUSION OCCURS AT 512 00:25:49,465 --> 00:25:59,475 THE TIP OF BASIDIA, AND THEN ME 513 00:25:59,475 --> 00:26:08,317 IOS IS AND MEITIC NUCLEI 514 00:26:08,317 --> 00:26:09,585 REMAIN -- BUD OUT THE SPECIES IN 515 00:26:09,585 --> 00:26:11,120 THE LONG CHAIN. 516 00:26:11,120 --> 00:26:12,588 THIS IS A SCANNING ELECTRON 517 00:26:12,588 --> 00:26:13,789 MICROSCOPY. 518 00:26:13,789 --> 00:26:15,658 SO COMPLETELY DIFFERENT 519 00:26:15,658 --> 00:26:18,594 MORPHOLOGY OF ASEXUAL AND SEXUAL 520 00:26:18,594 --> 00:26:24,166 STAGE OF CRYPTOCOCCUS. 521 00:26:24,166 --> 00:26:28,204 HOW DID IT AFFECT TAXONOMY? 522 00:26:28,204 --> 00:26:31,507 WELL, WE ALREADY SHOWED THAT 523 00:26:31,507 --> 00:26:34,276 THERE MUST BE TWO SPECIES. 524 00:26:34,276 --> 00:26:36,245 WE KNEW CRYPTOCOCCUS WAS 525 00:26:36,245 --> 00:26:39,415 HETEROGENEOUS BUT NOT QUITE AS 526 00:26:39,415 --> 00:26:39,715 TWO SPECIES. 527 00:26:39,715 --> 00:26:45,888 SO THE TWO SPECIES CONCEPT WAS 528 00:26:45,888 --> 00:26:48,724 EMERGED AFTER I SHOWED THIS 529 00:26:48,724 --> 00:26:50,459 PERFECT STATE. 530 00:26:50,459 --> 00:26:52,762 AND BEFORE THIS PERFECT STATE, 531 00:26:52,762 --> 00:26:54,530 CRYPTOCOCCUS WAS ALWAYS 532 00:26:54,530 --> 00:27:00,302 IDENTIFIED AS A SEROTYPE A OR 533 00:27:00,302 --> 00:27:00,936 DOB OR C. 534 00:27:00,936 --> 00:27:09,078 THIS IS ALL DR. BENNETT'S WORK. 535 00:27:09,078 --> 00:27:12,548 NOW WHEN I DID THE MATING, AD 536 00:27:12,548 --> 00:27:14,316 MATING TYPE PRODUCED THIS KIND 537 00:27:14,316 --> 00:27:18,821 OF SEXUAL STATE, WHEREAS B AND C 538 00:27:18,821 --> 00:27:20,056 SEROTYPE PRODUCE COMPLETELY 539 00:27:20,056 --> 00:27:30,599 DIFFERENT MORPHOLOGY OF SPORES. 540 00:27:30,833 --> 00:27:32,201 AND IT'S REALLY SURPRISING 541 00:27:32,201 --> 00:27:36,038 BECAUSE OF THE 20 STRAINS HE 542 00:27:36,038 --> 00:27:38,774 PICKED RANDOMLY, THERE WERE TWO 543 00:27:38,774 --> 00:27:41,043 MATING TYPES, AND WE'LL LOOK AT 544 00:27:41,043 --> 00:27:42,378 WHY I SAY THIS. 545 00:27:42,378 --> 00:27:44,547 SO IF YOU DISCOVER THE 546 00:27:44,547 --> 00:27:45,548 HETEROTHALLISM, HOW DID IT 547 00:27:45,548 --> 00:27:51,120 IMPACT THE FIELD? 548 00:27:51,120 --> 00:27:53,089 EPIDEMIOLOGICAL SIGNIFICANCE OF 549 00:27:53,089 --> 00:27:53,856 MATING TYPE. 550 00:27:53,856 --> 00:27:56,692 WE JUST THINK OH, THERE'S GOT TO 551 00:27:56,692 --> 00:28:01,130 BE META, METAL FA, HALF AND 552 00:28:01,130 --> 00:28:02,865 HALF, AMONG CLINICAL 553 00:28:02,865 --> 00:28:03,365 ENVIRONMENTAL SOILS. 554 00:28:03,365 --> 00:28:06,202 WHAT WE WERE SO SURPRISED WAS 555 00:28:06,202 --> 00:28:11,340 METAL FA WAS 99.8%, MATTA IS 556 00:28:11,340 --> 00:28:11,774 ONLY .2%. 557 00:28:11,774 --> 00:28:14,844 IF YOU THINK ABOUT THE 558 00:28:14,844 --> 00:28:17,146 PROBABILITY OF MYSELF GETTING 20 559 00:28:17,146 --> 00:28:21,517 STRAINS FROM DR. BENNETT IN 560 00:28:21,517 --> 00:28:24,353 WHICH TWO EYES LATS WERE META, 561 00:28:24,353 --> 00:28:26,122 CAN YOU IMAGINE, SO I 562 00:28:26,122 --> 00:28:28,090 CALCULATED, IT LOOKED LIKE 563 00:28:28,090 --> 00:28:30,359 PROBABILITY IS 1 IN 1500 564 00:28:30,359 --> 00:28:31,360 CHANCES. 565 00:28:31,360 --> 00:28:35,197 1 IN 1500. 566 00:28:35,197 --> 00:28:38,267 SO I WAS VERY LUCKY THAT HE 567 00:28:38,267 --> 00:28:39,468 SELECTED 20 ISOLATES AMONG WHICH 568 00:28:39,468 --> 00:28:42,104 THERE WERE TWO MATING TYPE A 569 00:28:42,104 --> 00:28:42,338 STRAIN. 570 00:28:42,338 --> 00:28:44,173 THAT WAS ALL HE HAD, TWO MATING 571 00:28:44,173 --> 00:28:49,979 TYPE A STRAIN OUT OF 1,000. 572 00:28:49,979 --> 00:28:52,681 CONSTRUCTION, CONGENIC SET OF 573 00:28:52,681 --> 00:28:53,883 MATTA AND MATT ALPHA, BECAUSE 574 00:28:53,883 --> 00:28:57,820 WHEN YOU SEE THAT MAT ALPHA 575 00:28:57,820 --> 00:28:58,487 PREDOMINATES, THEN YOU'RE 576 00:28:58,487 --> 00:29:03,092 CURIOUS ABOUT WHAT IS MATALPHA 577 00:29:03,092 --> 00:29:06,595 DIFFERENT FROM MATA AND WHY IS 578 00:29:06,595 --> 00:29:08,130 IT SO MUCH MORE COMMON? 579 00:29:08,130 --> 00:29:08,898 NEXT QUESTION OF COURSE YOU WILL 580 00:29:08,898 --> 00:29:10,866 ASK IS, WHAT IS THE ROLE OF 581 00:29:10,866 --> 00:29:15,004 MATING TYPE IN VIRULENCE? 582 00:29:15,004 --> 00:29:16,539 MAT ALPHA WAS FOUND TO BE 583 00:29:16,539 --> 00:29:18,841 SIGNIFICANTLY MORE VIRULENT THAN 584 00:29:18,841 --> 00:29:23,312 MATA IN THE ANIMAL MODEL. 585 00:29:23,312 --> 00:29:24,446 WE DON'T KNOW WHY, BECAUSE 586 00:29:24,446 --> 00:29:26,415 THERE'S JUST SO FEW MAT A OUT 587 00:29:26,415 --> 00:29:27,483 THERE. 588 00:29:27,483 --> 00:29:31,554 AND GENOMIC ANALYSIS OF MAT 589 00:29:31,554 --> 00:29:33,823 LOCI, AND EVOLUTION OF FUNGAL 590 00:29:33,823 --> 00:29:42,031 SEX CHROMOSOME PROJECT EMERGED. 591 00:29:42,031 --> 00:29:44,133 MITOCHONDRIA INHERITANCE, UNO 592 00:29:44,133 --> 00:29:46,202 PARENTAL INHERITANCE FORMAT A 593 00:29:46,202 --> 00:29:49,905 PARENT JUST LIKE MAMMALIAN CASE, 594 00:29:49,905 --> 00:29:51,774 THE MITOCHONDRIA COMES FROM THE 595 00:29:51,774 --> 00:29:54,176 MOTHER, BUT MITOCHONDRIA FROM 596 00:29:54,176 --> 00:29:57,012 CRYPTO COMES FROM MAT A ISOLATE. 597 00:29:57,012 --> 00:30:00,549 NOW, WHO BENEFITED FROM ALL THIS 598 00:30:00,549 --> 00:30:02,084 DISCOVERY OF HETEROTHALLISM AND 599 00:30:02,084 --> 00:30:06,222 BUILT THEIR CAREER ON THE MAT 600 00:30:06,222 --> 00:30:07,122 MATING? 601 00:30:07,122 --> 00:30:10,292 JOE HEITMAN IN DUKE, HE HAD ALL 602 00:30:10,292 --> 00:30:15,431 THIS COMPLETING THE MATING TYPE 603 00:30:15,431 --> 00:30:20,135 LOCI GENETIC MAP OF MATING TYPE 604 00:30:20,135 --> 00:30:21,971 LOCI. 605 00:30:21,971 --> 00:30:25,507 AND PHEROMONE SIGNALING CASCADE 606 00:30:25,507 --> 00:30:26,909 AND CELLULAR CIRCUITS REQUIRED 607 00:30:26,909 --> 00:30:27,576 FOR MATING. 608 00:30:27,576 --> 00:30:30,412 AND HE SPENT PROBABLY NO LESS 609 00:30:30,412 --> 00:30:36,151 THAN 10 YEARS TO ACCOMPLISH ALL 610 00:30:36,151 --> 00:30:37,519 THIS. 611 00:30:37,519 --> 00:30:39,488 AND X. LIN NOW IN GEORGIA, BUT 612 00:30:39,488 --> 00:30:41,790 SHE USED TO BE AT TEXAS A & M, 613 00:30:41,790 --> 00:30:43,759 SHE DISCOVERED THE SAME SEX 614 00:30:43,759 --> 00:30:47,763 MATING AMONG CRYPTOCOCCUS. 615 00:30:47,763 --> 00:30:51,200 AND PHEROMONE INDEPENDENT 616 00:30:51,200 --> 00:30:51,734 UNISEXUAL MATING. 617 00:30:51,734 --> 00:30:54,470 AND THEN DR. XU IN CANADA, HE'S 618 00:30:54,470 --> 00:30:57,973 THE ONE STUDIED THE MAT A 619 00:30:57,973 --> 00:30:58,874 DEPENDENT MITOCHONDRIAL 620 00:30:58,874 --> 00:30:59,742 INHERITANCE. 621 00:30:59,742 --> 00:31:03,245 SO ALL THESE PEOPLE BUILT THEIR 622 00:31:03,245 --> 00:31:06,515 CAREER BENEFITING FROM 623 00:31:06,515 --> 00:31:07,716 HETEROTHALLISM LIFECYCLE OF 624 00:31:07,716 --> 00:31:12,955 CRYPTOCOCCUS. 625 00:31:12,955 --> 00:31:15,257 THIS IS 2000, SO THIS IS LONG 626 00:31:15,257 --> 00:31:19,295 BEFORE CRY CRYPTOCOCCAL GENOME 627 00:31:19,295 --> 00:31:22,898 SEQUENCING WAS ESTABLISHED. 628 00:31:22,898 --> 00:31:25,434 WHEN MARVIN KAROS FROM GERMANY 629 00:31:25,434 --> 00:31:33,108 WAS WITH US, -- WAS TEACHING HOW 630 00:31:33,108 --> 00:31:35,144 TO ISOLATE THE MATING TYPES AND 631 00:31:35,144 --> 00:31:39,114 WE WERE SO SU SURPRISED THAT ITS 632 00:31:39,114 --> 00:31:41,383 AT LEAST 50KB LONG, AND IF YOU 633 00:31:41,383 --> 00:31:43,152 LOOK AT -- THERE'S ONLY TWO 634 00:31:43,152 --> 00:31:45,554 GENES AND THEY ARE LIKE LESS 635 00:31:45,554 --> 00:31:45,988 THAN 8KB. 636 00:31:45,988 --> 00:31:50,259 COMPARED TO THAT, 50KB AND ALL 637 00:31:50,259 --> 00:31:51,794 THE MAT KINASE CASCADE GENES 638 00:31:51,794 --> 00:31:54,530 WERE SITTING ON THE MAT LOCUS. 639 00:31:54,530 --> 00:32:00,636 AND AS I SAID, AND WHEN GENOME 640 00:32:00,636 --> 00:32:07,109 SEQUENCING WAS DONE IN 2004, 641 00:32:07,109 --> 00:32:08,510 HEITMAN AT DUKE COMPLETED AND 642 00:32:08,510 --> 00:32:10,379 BOTH MATING TYPE, THE MATING 643 00:32:10,379 --> 00:32:17,386 TYPE LOCUS WAS 100KB OR 117KB, 644 00:32:17,386 --> 00:32:24,994 IT'S ALMOST SEX CHROMOSOME. 645 00:32:24,994 --> 00:32:27,329 ABOUT PHYLOGENY, HOW DID THEY 646 00:32:27,329 --> 00:32:30,933 BENEFIT, HOW DID THE AREA OF 647 00:32:30,933 --> 00:32:36,972 PHYLOGENY BENEFIT FROM 648 00:32:36,972 --> 00:32:37,306 HYPOTHALLISM? 649 00:32:37,306 --> 00:32:38,607 IF YOU SEE THIS MORPHOLOGY, THIS 650 00:32:38,607 --> 00:32:39,808 IS THE SEXUAL STATE. 651 00:32:39,808 --> 00:32:42,544 NOW WE CAN COMPARE ABOUT THE 652 00:32:42,544 --> 00:32:45,280 REMAINING FUNGAL SPECIES IN THE 653 00:32:45,280 --> 00:32:46,582 KINGDOM FUNGI. 654 00:32:46,582 --> 00:32:51,954 THE NEAREST ONE WAS THIS 655 00:32:51,954 --> 00:32:57,092 STRUCTURE, PRODUCED BY TREMELLA. 656 00:32:57,092 --> 00:33:02,898 THAT IS WOOD EAR MUSHROOM. 657 00:33:02,898 --> 00:33:04,867 THEY ALL GROW ON TREE BARK. 658 00:33:04,867 --> 00:33:12,908 SO FINALLY, WE FL WE NOTICE THIS 659 00:33:12,908 --> 00:33:14,209 CRYPTO COCCUS SHOULD BELONG TO 660 00:33:14,209 --> 00:33:16,612 THE TREMELLA GROUP. 661 00:33:16,612 --> 00:33:19,481 WHEN DNA SEQUENCING BECAME 662 00:33:19,481 --> 00:33:20,549 AVAILABLE, PHYLOGENETIC TREE 663 00:33:20,549 --> 00:33:24,286 BASED ON ITS INDEED SHOWED THAT 664 00:33:24,286 --> 00:33:27,089 CRYPTOCOCCUS GROUP AND THEN 665 00:33:27,089 --> 00:33:29,191 TREMELLA SPECIES WERE IN THE 666 00:33:29,191 --> 00:33:32,261 SAME ORDER. 667 00:33:32,261 --> 00:33:36,532 SO NOW WE COULD PLACE THE 668 00:33:36,532 --> 00:33:44,073 CRYPTOCOCCUS IN THE TREMELLALES 669 00:33:44,073 --> 00:33:44,740 IN MICROBIOTA. 670 00:33:44,740 --> 00:33:46,275 BECAUSE OF ALL THIS DIFFERENCE, 671 00:33:46,275 --> 00:33:50,646 FAY LOW GENPHYLOGENETIC STUDY WD 672 00:33:50,646 --> 00:33:52,481 BY MANY PEOPLE AND EVENTUALLY 673 00:33:52,481 --> 00:34:00,689 FOUND THAT CRYPTOCOCCUS GATT 674 00:34:00,689 --> 00:34:06,595 GATTII -- THIS PERSON, 675 00:34:06,595 --> 00:34:09,998 DR. MITCHELL, IS IN DUKE, AND 676 00:34:09,998 --> 00:34:14,903 MARIANA IS IN ITALY, AND 677 00:34:14,903 --> 00:34:17,106 BOEKHOUT IS IN POLAND, AND MEYER 678 00:34:17,106 --> 00:34:18,307 WAS IN AUSTRALIA. 679 00:34:18,307 --> 00:34:20,676 SO IT'S LIKE WHOLE INTERNATIONAL 680 00:34:20,676 --> 00:34:23,979 TEAMS START STUDYING THE 681 00:34:23,979 --> 00:34:34,156 PHYLOGENY. 682 00:34:48,003 --> 00:34:49,872 SO THE FIRST GOAL WAS DISCOVERED 683 00:34:49,872 --> 00:34:51,106 IN THE FULL LIFECYCLE. 684 00:34:51,106 --> 00:34:56,044 NOW THAT'S DONE, AND THE SECOND 685 00:34:56,044 --> 00:34:57,679 GOAL WAS TO FIND THE VIRULENCE 686 00:34:57,679 --> 00:34:58,013 FACTOR. 687 00:34:58,013 --> 00:34:59,548 AND HOW DO YOU FIND THE VIEWER 688 00:34:59,548 --> 00:35:03,152 LANCE FACTOR? 689 00:35:03,152 --> 00:35:03,886 VIRULENCE FACTOR? 690 00:35:03,886 --> 00:35:06,421 SO WHAT I DID WAS A DICHOTOMOUS 691 00:35:06,421 --> 00:35:07,422 DIAGNOSTIC APPROACH OF -- THAT 692 00:35:07,422 --> 00:35:09,191 TIME THERE WERE ONLY 27 SPECIES 693 00:35:09,191 --> 00:35:11,994 IN THE GENUS CRYPT COCCUS. 694 00:35:11,994 --> 00:35:15,831 SO THEIR ABILITY TO UTILIZE 695 00:35:15,831 --> 00:35:19,001 CARBOHYDRATES AND NITROGEN 696 00:35:19,001 --> 00:35:22,404 SOURCE AND THE GROWTH ABILITY AT 697 00:35:22,404 --> 00:35:25,140 37 DEGREE, AND THEN THE 698 00:35:25,140 --> 00:35:27,209 PHENOTYPE OF COLONY THAT WAS 699 00:35:27,209 --> 00:35:33,549 GROWN ON THE MEDIA THAT HAS 700 00:35:33,549 --> 00:35:34,016 DIPHENOLIC COMPOUNDS. 701 00:35:34,016 --> 00:35:36,852 SO I MADE THIS DIAGRAM, 702 00:35:36,852 --> 00:35:44,293 DICHOTOMOUS DIAGRAM, AND WHAT 703 00:35:44,293 --> 00:35:51,066 STOOD OUT -- I HAVE CIRCLED HERE 704 00:35:51,066 --> 00:35:52,801 BUT IT DOESN'T SHOW. 705 00:35:52,801 --> 00:35:58,173 AT THE TOP LEFT, IT SAYS COLONY 706 00:35:58,173 --> 00:36:03,946 IS BROWN, LECTOSE IS 707 00:36:03,946 --> 00:36:04,947 NON-UTILIZED AND GROWTH AT 708 00:36:04,947 --> 00:36:05,414 37 DEGREE. 709 00:36:05,414 --> 00:36:10,852 I LOOKED AT ALL 27 SPECIES, 710 00:36:10,852 --> 00:36:12,821 CRYPTOCOCCUS WAS UNIQUE 711 00:36:12,821 --> 00:36:13,488 DISTINGUISHING CHARACTER AS 712 00:36:13,488 --> 00:36:20,262 THIS. 713 00:36:20,262 --> 00:36:23,865 WHY BROWN COLONY ON THE 714 00:36:23,865 --> 00:36:25,834 DIPHENOLIC MEDIA? 715 00:36:25,834 --> 00:36:29,004 PROFESSOR STIVE, WHO USED TO BE 716 00:36:29,004 --> 00:36:31,406 WORKING AT ROBERT KOCH'S 717 00:36:31,406 --> 00:36:33,041 INSTITUTE IN BERLIN, AT THAT 718 00:36:33,041 --> 00:36:35,143 TIME, OF COURSE, THE ECOLOGICAL 719 00:36:35,143 --> 00:36:40,716 NICHE OF CRYPTOCOCCUS NEOFOR 720 00:36:40,716 --> 00:36:42,451 MENS BEST KNOWN IS THE PIGEON 721 00:36:42,451 --> 00:36:43,885 DROPPING, SO HE THINK MAYBE 722 00:36:43,885 --> 00:36:46,388 PIGEONS EAT THE BIRD SEED AND 723 00:36:46,388 --> 00:36:48,490 SOMEHOW BIRD SEED IS SELECTIVELY 724 00:36:48,490 --> 00:36:49,691 LETTING CRYPTOCOCCUS GROW MUCH 725 00:36:49,691 --> 00:36:50,125 BETTER. 726 00:36:50,125 --> 00:36:51,994 SO HE EXTRACTED THE BIRD SEED, 727 00:36:51,994 --> 00:36:55,597 PUT IT INTO THE AUGER, AND THEN 728 00:36:55,597 --> 00:37:00,502 PLATED THE CLINICAL YEAST, ALL 729 00:37:00,502 --> 00:37:02,371 THE CANDIDA SPECIES REMAIN WHITE 730 00:37:02,371 --> 00:37:06,208 AND KN NEOFORM CRYPTOCOCCUS IS E 731 00:37:06,208 --> 00:37:07,943 ONLY ONE THAT PRODUCED THE ROUND 732 00:37:07,943 --> 00:37:11,446 COLONY, THE MELANIN. 733 00:37:11,446 --> 00:37:14,416 AND SO REMEMBER 37 DEGREE 734 00:37:14,416 --> 00:37:15,717 GROWTH, MELANIN INFORMATION ON 735 00:37:15,717 --> 00:37:20,422 MEDIA, AND THE NEGATIVE 736 00:37:20,422 --> 00:37:27,095 UTILIZATION LECTOSE, SO THE NEXT 737 00:37:27,095 --> 00:37:29,264 THING WE DID WAS MADE A MU TAN, 738 00:37:29,264 --> 00:37:30,465 AT THAT TIME OF COURSE THERE'S 739 00:37:30,465 --> 00:37:32,000 NO GENETIC MANIPULATION, YOU DO 740 00:37:32,000 --> 00:37:34,636 UV RADIATION AND GET THE MELANIN 741 00:37:34,636 --> 00:37:35,837 NEGATIVE AND THEN LEAVE THE 742 00:37:35,837 --> 00:37:38,473 MELANIN POSITIVE PARENT, CROSS 743 00:37:38,473 --> 00:37:45,147 THEM, AND THEN ISOLATED ALL THE 744 00:37:45,147 --> 00:37:46,214 F1 PROGENY AND THERE'S NO 745 00:37:46,214 --> 00:37:48,950 QUESTION IT'S MAN DALIAN 746 00:37:48,950 --> 00:37:49,951 INHERITANCE OF THE MELANIN FOR 747 00:37:49,951 --> 00:37:51,753 PLAITION EXACTLY OUT OF 25 748 00:37:51,753 --> 00:37:54,156 COLONIES, 12 TO 13. 749 00:37:54,156 --> 00:37:56,858 AND THEN 37 DEGREE GROWTH. 750 00:37:56,858 --> 00:37:59,227 AND THEN CAPSULE. 751 00:37:59,227 --> 00:38:04,166 THESE WERE BY FAR THE ESSENTIAL 752 00:38:04,166 --> 00:38:08,270 VIRULENCE FACTOR OF CRYPTOCO 753 00:38:08,270 --> 00:38:10,505 CRYPTOCOCCUS. 754 00:38:10,505 --> 00:38:12,274 AND TWO FELLOWS WORKED ON THESE 755 00:38:12,274 --> 00:38:14,776 PROJECTS, DR. POLACHECK AND JUDY 756 00:38:14,776 --> 00:38:16,178 ROSE, BOTH OF THEM ARE RETIRED 757 00:38:16,178 --> 00:38:20,148 NOW, BUT THIS CHART SHOWS THE 758 00:38:20,148 --> 00:38:29,091 WILD TYPE AND THEN REV REVERTAN, 759 00:38:29,091 --> 00:38:33,028 BUT MELANIN NEGATIVE OR CAPSULE 760 00:38:33,028 --> 00:38:35,997 NEGATIVES ARE ALMOST AVIRULENT 761 00:38:35,997 --> 00:38:36,765 DEPENDING ON THE STRAIN. 762 00:38:36,765 --> 00:38:40,602 AND THIS IS A MOUSE STUDY. 763 00:38:40,602 --> 00:38:45,474 SO WE ESTABLISHED THAT BUY YOE 764 00:38:45,474 --> 00:38:49,378 CHEMICAL STEPS IN MELANOGENESIS 765 00:38:49,378 --> 00:38:51,546 DESCRIBED AND INVOLVEMENT OF 766 00:38:51,546 --> 00:38:52,881 ONLY ONE ENZYME, BECAUSE IT WAS 767 00:38:52,881 --> 00:38:54,616 CLEAR THAT ONLY ONE ENZYME WAS 768 00:38:54,616 --> 00:38:56,685 INVOLVED. 769 00:38:56,685 --> 00:39:00,522 AND THAT WAS LACCASE, WE CALLED 770 00:39:00,522 --> 00:39:02,724 PHENOL OXIDASE. 771 00:39:02,724 --> 00:39:04,459 THAT POSITION OF MELANIN IN THE 772 00:39:04,459 --> 00:39:09,264 CELL WALL WE NOTICED, AND THEN 773 00:39:09,264 --> 00:39:11,800 PETER WILLIAMSON, WHO CAME HERE, 774 00:39:11,800 --> 00:39:15,404 HE'S SITTING HERE, BUT HE CAME 775 00:39:15,404 --> 00:39:16,938 HERE AS A CLINICAL FELLOW, AND 776 00:39:16,938 --> 00:39:19,674 THE FIRST 20 YEARS OF HIS CAREER 777 00:39:19,674 --> 00:39:23,278 WAS TOTALLY FOCUSED ON HOW 778 00:39:23,278 --> 00:39:25,580 CRYPTOCOCCUS PRODUCED MELANIN. 779 00:39:25,580 --> 00:39:29,384 AND HOW IS IT SYNTHESIZED, WHAT 780 00:39:29,384 --> 00:39:30,719 KINDS OF GENES ARE REGULATED, 781 00:39:30,719 --> 00:39:32,888 HOW IS IT BEING 782 00:39:32,888 --> 00:39:33,655 SHIPPED/TRANSPORTED TO THE CELL 783 00:39:33,655 --> 00:39:35,524 WALL AND EVENTUALLY SECRETED? 784 00:39:35,524 --> 00:39:38,260 ALL OF THESE PICTURES, THERE ARE 785 00:39:38,260 --> 00:39:40,996 JUST TONS OF GENETIC FACTORS 786 00:39:40,996 --> 00:39:42,664 INVOLVED TO MAKE THE MELANIN 787 00:39:42,664 --> 00:39:45,133 FORMATION. 788 00:39:45,133 --> 00:39:47,903 SO I'M GLAD PETER STARTED 789 00:39:47,903 --> 00:39:50,272 WORKING ON MELANIN AS SOON AS WE 790 00:39:50,272 --> 00:39:57,412 KNEW IT WAS A VIRULENCE FACTOR. 791 00:39:57,412 --> 00:40:00,148 AND THEN, RIGHT NOW AT HOPKINS, 792 00:40:00,148 --> 00:40:03,652 PREVIOUSLY HE WAS AT NEW YORK, 793 00:40:03,652 --> 00:40:06,254 HE PRODUCED MELANIN GHOST MADE 794 00:40:06,254 --> 00:40:16,898 FROM MELANIZED CRY -- MELANIN 795 00:40:19,568 --> 00:40:20,802 STAINED GHOST, AND THE 796 00:40:20,802 --> 00:40:24,105 INTERESTING PART IS THIS 797 00:40:24,105 --> 00:40:25,507 ANTIBODY IS CURRENTLY IN 798 00:40:25,507 --> 00:40:28,043 EVALUATION FOR THE TREATMENT OF 799 00:40:28,043 --> 00:40:31,746 HUMAN MELANOMA, BECAUSE HUMAN 800 00:40:31,746 --> 00:40:37,752 MELANIN, CRYPT CO CRYPTOCOCCAL N 801 00:40:37,752 --> 00:40:39,721 ARE BOTH U MELANINS. 802 00:40:39,721 --> 00:40:42,557 DISCOVERY OF THIS CGV DIAGNOSTIC 803 00:40:42,557 --> 00:40:43,892 MEDIA AND THE IMPACT ON THE 804 00:40:43,892 --> 00:40:49,564 PROGRESS OF CRYPT CO CRYPTOCOCCY 805 00:40:49,564 --> 00:40:50,332 AND EPIDEMIOLOGY. 806 00:40:50,332 --> 00:40:52,634 BEFORE MY MEDIA WAS MADE, WHAT 807 00:40:52,634 --> 00:40:56,238 WE KNEW WAS JUST ABOUT THIS 808 00:40:56,238 --> 00:40:59,074 NATURAL SOURCE OF CRYPTOCOCCUS 809 00:40:59,074 --> 00:41:01,042 WAS PIGEON DROPPINGS, AND THEN 810 00:41:01,042 --> 00:41:08,617 THE SOIL CONTAMINATED BY AVEEN 811 00:41:08,617 --> 00:41:10,151 GUANOS. 812 00:41:10,151 --> 00:41:13,188 SECOND SPECIES, CRYPTOCOCCUS 813 00:41:13,188 --> 00:41:14,723 GAATII WAS NEVER ISOLATED FROM 814 00:41:14,723 --> 00:41:17,459 ENVIRONMENT UNTIL 1990. 815 00:41:17,459 --> 00:41:24,032 BECAUSE GAATII AND NEOFORMANS 816 00:41:24,032 --> 00:41:27,202 WERE DISCOVERED ONE YEAR APART, 817 00:41:27,202 --> 00:41:28,837 1994 AND ONE IN 1995, SO IT'S 818 00:41:28,837 --> 00:41:30,505 ALMOST LIKE 100 YEARS THAT WE 819 00:41:30,505 --> 00:41:32,874 DIDN'T KNOW WHERE GATTII WAS 820 00:41:32,874 --> 00:41:37,579 COMING FROM. 821 00:41:37,579 --> 00:41:39,147 OUR DIAGNOSTIC MEDIA BECAME 822 00:41:39,147 --> 00:41:42,617 INSTRUMENTAL FOR THE STUDY OF 823 00:41:42,617 --> 00:41:44,920 C.GATTII FROM EUCALYPTUS TREE IN 824 00:41:44,920 --> 00:41:52,894 1990 WHICH LED TO RESEARCH FOR 825 00:41:52,894 --> 00:41:54,963 ARBORIA FORM, BECAUSE TREMELLA 826 00:41:54,963 --> 00:41:56,631 AND CRYPTOCOCCUS ARE VERY 827 00:41:56,631 --> 00:41:57,732 CLOSELY RELATED. 828 00:41:57,732 --> 00:41:59,367 TREMELLA GROWS ON TREE BARK AND 829 00:41:59,367 --> 00:42:01,303 WHY NOT CRYPTOCOCCUS? 830 00:42:01,303 --> 00:42:03,405 BUT EVERYONE WAS FOCUSING ON THE 831 00:42:03,405 --> 00:42:04,272 PIGEON DROPPINGS. 832 00:42:04,272 --> 00:42:08,143 THAT'S WHY WE NEVER FOUND, UNTIL 833 00:42:08,143 --> 00:42:08,343 1990. 834 00:42:08,343 --> 00:42:12,147 SO THIS IS IN MEDIA. 835 00:42:12,147 --> 00:42:14,549 NEOFORMANS DOES NOT UTILIZE -- 836 00:42:14,549 --> 00:42:23,758 CANNOT GROW IN CGB -- WHEREAS 837 00:42:23,758 --> 00:42:25,493 C.GATTII BECOMES BLUE WITHIN 24 838 00:42:25,493 --> 00:42:26,361 HOURS. 839 00:42:26,361 --> 00:42:30,532 WE'WHILE USING THIS MEDIA, FELLW 840 00:42:30,532 --> 00:42:33,702 IN AUSTRALIA, DAVID ELLIS, HE 841 00:42:33,702 --> 00:42:39,274 SAMPLED EUCALYPTUS TREE DROP THE 842 00:42:39,274 --> 00:42:41,142 LEAVES AND TREE BARKS SWABBED 843 00:42:41,142 --> 00:42:43,979 AND TONS OF YEAST WERE ISOLATED, 844 00:42:43,979 --> 00:42:47,148 BUT HE HAD TO LOOK FOR GATTII. 845 00:42:47,148 --> 00:42:49,884 HE USED MY MEDIA, AND HE 846 00:42:49,884 --> 00:42:57,125 ISOLATED THE FIRST GATTII FROM 847 00:42:57,125 --> 00:42:58,660 EUCALYPTUS TREE, AND HUNDREDS OF 848 00:42:58,660 --> 00:43:01,262 OTHER PEOPLE STARTED LOOKING FOR 849 00:43:01,262 --> 00:43:02,897 TREES, OF COURSE. 850 00:43:02,897 --> 00:43:04,633 SO THIS AN ECOLOGICAL IMPACT, 851 00:43:04,633 --> 00:43:12,140 BUT WHAT ABOUT EPIDEMIOLOGY? 852 00:43:12,140 --> 00:43:13,842 C. NEOFORMANS, AT LEAST 70% OF 853 00:43:13,842 --> 00:43:16,144 THEM ARE FROM HIV-POSITIVE 854 00:43:16,144 --> 00:43:17,879 PATIENT, CLINICAL SOURCE. 855 00:43:17,879 --> 00:43:22,484 AND THEN ONLY ABOUT 25% COMES 856 00:43:22,484 --> 00:43:22,917 FROM OTHERS. 857 00:43:22,917 --> 00:43:24,786 AS I SAID, WHAT THE RISK FACTOR 858 00:43:24,786 --> 00:43:28,056 IN THE BEGINNING. 859 00:43:28,056 --> 00:43:29,324 BUT INTERESTINGLY, IF YOU LOOK 860 00:43:29,324 --> 00:43:31,893 AT THE GATTII, VERY FEW COMES 861 00:43:31,893 --> 00:43:34,729 FROM AIDS PATIENTS, AND THE 862 00:43:34,729 --> 00:43:37,132 REMAINDERS OFTEN SAY PREVIOUSLY 863 00:43:37,132 --> 00:43:37,699 HEALTHY PATIENTS. 864 00:43:37,699 --> 00:43:41,503 SO THEY THOUGHT IT WAS AN 865 00:43:41,503 --> 00:43:42,170 IMMUNOCOMPETENT PATIENT, UNTIL 866 00:43:42,170 --> 00:43:45,106 WE FOUND THAT GM CSF AUTO 867 00:43:45,106 --> 00:43:46,441 ANTIBODY WAS A RISK FACTOR, 868 00:43:46,441 --> 00:43:48,943 WHICH HAS NEVER BEEN TESTED IN 869 00:43:48,943 --> 00:43:52,113 THE LABORATORY WHEN THEY WERE 870 00:43:52,113 --> 00:43:57,485 LOOKING FOR PATIENTS' 871 00:43:57,485 --> 00:43:59,554 IMMUNOLOGICAL -- SO GATTII WAS 872 00:43:59,554 --> 00:44:04,259 FOUND TO INFECT PREVIOUSLY 873 00:44:04,259 --> 00:44:05,460 HEALTHY PATIENTS, SO 874 00:44:05,460 --> 00:44:07,095 EPIDEMIOLOGY AND ECOLOGY BOTH 875 00:44:07,095 --> 00:44:10,932 DEVELOP THE VERY FAST AFTER THIS 876 00:44:10,932 --> 00:44:14,302 MEDIA HAS BEEN FORMULATED. 877 00:44:14,302 --> 00:44:16,271 NOW I LOOKED AT ALL THE TREES 878 00:44:16,271 --> 00:44:18,273 FROM WHICH THE CRYPTOCOCCUS 879 00:44:18,273 --> 00:44:20,775 COMES FROM BUT I JUST PICKED 12 880 00:44:20,775 --> 00:44:25,714 HERE BECAUSE THESE NAMES LIKE 881 00:44:25,714 --> 00:44:31,720 ACACIA, PINE AND OAK TREE AND 882 00:44:31,720 --> 00:44:32,721 PLATANUS, ALMOST EVERYONE KNOWS 883 00:44:32,721 --> 00:44:33,488 WHAT THEY ARE. 884 00:44:33,488 --> 00:44:35,390 BUT THERE ARE ABOUT MORE THAN 885 00:44:35,390 --> 00:44:40,395 100 TREES THAT GATTII AND 886 00:44:40,395 --> 00:44:44,666 NEOFORMANS ARE INHABITING. 887 00:44:44,666 --> 00:44:52,140 AND BY 1985, THAT PCR BECAME 888 00:44:52,140 --> 00:44:53,108 AVAILABLE. 889 00:44:53,108 --> 00:44:54,676 AND ALL OF MY WORK SO FAR HAS 890 00:44:54,676 --> 00:44:58,680 BEEN CLASSICAL, AND THEN THE 891 00:44:58,680 --> 00:45:00,081 HANDWRITING ON THE WALL OF MY 892 00:45:00,081 --> 00:45:03,618 LAB, KEEP ON SHOWING THAT YOU 893 00:45:03,618 --> 00:45:05,353 NEED TO CHANGE THE RESEARCH 894 00:45:05,353 --> 00:45:06,321 DIRECTION. 895 00:45:06,321 --> 00:45:07,655 REORIENT YOUR DIRECTION TO 896 00:45:07,655 --> 00:45:12,360 MOLECULAR GENETICS. 897 00:45:12,360 --> 00:45:14,429 IT WAS VERY INCONVENIENT, ALMOST 898 00:45:14,429 --> 00:45:20,769 SCARY TO STEP OUT OF MY COMFORT 899 00:45:20,769 --> 00:45:25,673 ZONE, BUT I DECIDED TO CHANGE 900 00:45:25,673 --> 00:45:27,142 IT, THAT WAS IN 1985. 901 00:45:27,142 --> 00:45:28,877 AT THAT TIME I WAS ALREADY EARLY 902 00:45:28,877 --> 00:45:33,815 50s, AND IT WAS A REALLY HARD 903 00:45:33,815 --> 00:45:40,488 DECISION I HAD TO MAKE. 904 00:45:40,488 --> 00:45:42,857 SO WHAT DID I DO AFTER CHANGING 905 00:45:42,857 --> 00:45:45,393 THE DIRECTION? 906 00:45:45,393 --> 00:45:46,928 FIRST THING I DID WAS DEVELOPING 907 00:45:46,928 --> 00:45:50,231 THE MOLECULAR TECHNIQUE OF 908 00:45:50,231 --> 00:45:52,167 CRYPTOCOCCUS AND COME UP WITH A 909 00:45:52,167 --> 00:45:58,540 TRANSFORMATION METHOD. 910 00:45:58,540 --> 00:46:01,476 AND THEN THE VIRULENCE FACTOR, 911 00:46:01,476 --> 00:46:03,878 MOLECULAR KOCH POSTULATE WAS 912 00:46:03,878 --> 00:46:04,212 FULL FIELD. 913 00:46:04,212 --> 00:46:05,413 SO I'M GOING TO TALK ABOUT THIS 914 00:46:05,413 --> 00:46:08,049 TOO BECAUSE THIS, TOO, COMES 915 00:46:08,049 --> 00:46:09,818 AFTER MY REORIENTATION, AND I 916 00:46:09,818 --> 00:46:18,226 HAVE NO TIME TO GO THROUGH HERE. 917 00:46:18,226 --> 00:46:19,894 SO WHAT DID I DO? 918 00:46:19,894 --> 00:46:23,264 I ENROLLED FOR EACH MOLECULAR 919 00:46:23,264 --> 00:46:24,699 BIOLOGY COURSE OFFERED BY 920 00:46:24,699 --> 00:46:26,201 COLDSPRING HARBOR AND I LEFT 921 00:46:26,201 --> 00:46:27,335 HOME TO STAY THREE WEEKS IN THE 922 00:46:27,335 --> 00:46:30,271 LAB, AND I FOUND I WAS THE ONLY 923 00:46:30,271 --> 00:46:32,273 ONE WHO ARE IN 50s, THE REST 924 00:46:32,273 --> 00:46:34,309 OF THEM WERE IN THEIR 20s AND 925 00:46:34,309 --> 00:46:40,348 30s AT THE MOST. 926 00:46:40,348 --> 00:46:42,884 SO DEVELOPED MOLECULAR TOOLS 927 00:46:42,884 --> 00:46:48,556 UPON ARRIVAL, AND WORKED IN 928 00:46:48,556 --> 00:46:50,425 COLLABORATION WITH JEFF EDMAN 929 00:46:50,425 --> 00:46:51,092 FROM UC SAN FRANCISCO. 930 00:46:51,092 --> 00:46:52,460 NOW WE HAD DEVELOPED 931 00:46:52,460 --> 00:46:55,029 TRANSFORMATION TECHNIQUE 932 00:46:55,029 --> 00:46:57,432 DEVELOPED. 933 00:46:57,432 --> 00:47:07,575 AND THAT MADE US TO DO MOLECU 934 00:47:07,575 --> 00:47:09,143 MOLECULAR -- POSTULATE FOR 935 00:47:09,143 --> 00:47:09,611 VIRULENCE FACTOR. 936 00:47:09,611 --> 00:47:14,415 AND BY DOING CLONING CAPSULE 937 00:47:14,415 --> 00:47:17,252 GENES AND TRANSFORMING CAP MINUS 938 00:47:17,252 --> 00:47:19,888 MUTANT, CAP POSITIVE DISRUPTION 939 00:47:19,888 --> 00:47:29,197 AND ALL OF THESE, MADE -- JUN 940 00:47:29,197 --> 00:47:30,732 CHANG, STAFF SCIENTIST IN OUR 941 00:47:30,732 --> 00:47:34,335 LAB, TOOK THIS PROJECT. 942 00:47:34,335 --> 00:47:36,738 AND USED THE CAP MINUS MUTANT, 943 00:47:36,738 --> 00:47:40,875 IT LOOKED LIKE -- YOU DON'T SEE 944 00:47:40,875 --> 00:47:42,410 ANY CAP -- COLONY IS VERY DRY, 945 00:47:42,410 --> 00:47:48,216 SO WHEN HE COMPLEMENTED WITH THE 946 00:47:48,216 --> 00:47:50,418 GENOMIC DNA, IT BECAME NOT ONLY 947 00:47:50,418 --> 00:47:53,488 CAPSID POSITIVE OF -- COLONY, 948 00:47:53,488 --> 00:47:56,858 BUT THE MOST IMPORTANT PART IS 949 00:47:56,858 --> 00:47:58,826 ALL THIS VIRULENCE CAME BACK. 950 00:47:58,826 --> 00:48:02,096 THE GREENS ARE THE CAP MINUS 951 00:48:02,096 --> 00:48:02,964 STRAINS, THERE'S FOUR DIFFERENT 952 00:48:02,964 --> 00:48:05,066 STRAINS. 953 00:48:05,066 --> 00:48:07,802 FOUR DIFFERENT MUTANTS. 954 00:48:07,802 --> 00:48:11,072 AND THEN THE RED IS THE WILD 955 00:48:11,072 --> 00:48:15,176 TYPE, AND BLUE IS THE ONE WHICH 956 00:48:15,176 --> 00:48:16,778 IS COMPLEMENTED THIS MUTANT, AND 957 00:48:16,778 --> 00:48:23,451 THEY ALL BECAME VIRULENT. 958 00:48:23,451 --> 00:48:27,488 SO AFTER THIS HUGE CHANGE 959 00:48:27,488 --> 00:48:29,290 OCCURRED, CRYPTOCOCCUS RESEARCH 960 00:48:29,290 --> 00:48:31,292 FIELD EXPERIENCES SUDDEN 961 00:48:31,292 --> 00:48:33,394 TURNABOUT FROM MID 1990s. 962 00:48:33,394 --> 00:48:34,729 WHY? 963 00:48:34,729 --> 00:48:40,268 BECAUSE AIDS CAME ON, AND THEN 964 00:48:40,268 --> 00:48:46,507 LOW LEVEL OF LAB HAZARD, SO SHEY 965 00:48:46,507 --> 00:48:47,709 SHOULDN'T BE AFRAID TO MOVE IN 966 00:48:47,709 --> 00:48:49,811 FROM OTHER FIELD, AND GENETIC 967 00:48:49,811 --> 00:48:52,113 AMENABILITY DUE TO THE CROSSING 968 00:48:52,113 --> 00:48:54,849 AND TRANSFORMATION AND ALL OF 969 00:48:54,849 --> 00:48:56,718 THIS, AVAILABILITY OF MOLECULAR 970 00:48:56,718 --> 00:48:59,988 TOOLS, AND CONFIRMED VIRULENCE 971 00:48:59,988 --> 00:49:02,423 FACTORS SO THEY CAN GO AFTER 972 00:49:02,423 --> 00:49:05,493 VIRULENCE, AND GENETIC 973 00:49:05,493 --> 00:49:06,794 DIVERSITY, AND EXCELLENT ANIMAL 974 00:49:06,794 --> 00:49:07,128 MODELS. 975 00:49:07,128 --> 00:49:10,298 BECAUSE OF ALL THESE, PEOPLE 976 00:49:10,298 --> 00:49:16,738 START MOVING IN, INFLUX FROM ALL 977 00:49:16,738 --> 00:49:18,072 THESE AREAS WITH A STRONG 978 00:49:18,072 --> 00:49:21,442 BACKGROUND IN MOLECULAR GENETICS 979 00:49:21,442 --> 00:49:24,846 OF SACK ROW MY CYST OR OTHER 980 00:49:24,846 --> 00:49:27,482 MICROORGANISM. 981 00:49:27,482 --> 00:49:31,085 AND GLYCOBIOLOGY, BECAUSE THE 982 00:49:31,085 --> 00:49:32,754 CAPSULE BEING THE MAY YOUR 983 00:49:32,754 --> 00:49:34,722 VIRULENCE FACTOR AND KNEW VERY 984 00:49:34,722 --> 00:49:37,558 LITTLE ABOUT CAPSULE SYNTHESIS 985 00:49:37,558 --> 00:49:40,294 MICROBIOLOGY CAME IN AND 986 00:49:40,294 --> 00:49:41,462 EVOLUTION BIOLOGISTS BECAUSE OF 987 00:49:41,462 --> 00:49:45,967 THE DIVERSITY OF CRYPTOCOCCAL 988 00:49:45,967 --> 00:49:49,404 POPULATION, AND GENETICS 989 00:49:49,404 --> 00:49:51,472 BIOINFORMATICS AND CELLULAR AND 990 00:49:51,472 --> 00:49:53,207 MOLECULAR IMMUNOLOGY ESPECIALLY 991 00:49:53,207 --> 00:49:58,679 BECAUSE EXCELLENT ANIMAL MODELS. 992 00:49:58,679 --> 00:50:02,417 EVENTUALLY BY YEAR 2011, 993 00:50:02,417 --> 00:50:04,519 CRYPTOCOCCUS BECAME A MODEL 994 00:50:04,519 --> 00:50:08,890 YEAST FOR THE STUDY OF FUNGAL 995 00:50:08,890 --> 00:50:10,525 PATHOGENICITY AND PUBLISHED A 996 00:50:10,525 --> 00:50:13,828 BOOK BY ASN. 997 00:50:13,828 --> 00:50:19,634 I TRIED TO HAVE AS MANY PAPERS 998 00:50:19,634 --> 00:50:21,602 PICTURE BUT THIS IS ALL I COULD 999 00:50:21,602 --> 00:50:21,936 GET. 1000 00:50:21,936 --> 00:50:24,238 BY THE YEAR 2017, ALL OF THESE 1001 00:50:24,238 --> 00:50:26,641 PEOPLE I CALL A VILLAGE, PLUS 1002 00:50:26,641 --> 00:50:32,647 THERE WERE ABOUT 400 AUTHORS, 1003 00:50:32,647 --> 00:50:33,247 CO-AUTHORS/COLLABORATORS, AND 1004 00:50:33,247 --> 00:50:35,716 WORKED SO THAT I COULD 1005 00:50:35,716 --> 00:50:40,121 ACCOMPLISH TO THIS DAY. 1006 00:50:40,121 --> 00:50:40,555 CONCLUSION. 1007 00:50:40,555 --> 00:50:42,757 NIAID PLAYED A KEY ROLE IN 1008 00:50:42,757 --> 00:50:45,259 LAYING THE FOUNDATION FOR 1009 00:50:45,259 --> 00:50:46,928 PROGRESS IN THE CRYPTOCOCCAL 1010 00:50:46,928 --> 00:50:53,801 RESEARCH STUDY IN 1975, AND 1011 00:50:53,801 --> 00:50:55,136 RE-ORIENTING THE RESEARCH 1012 00:50:55,136 --> 00:50:59,841 DIRECTION TO STAY ALIGNED WITH 1013 00:50:59,841 --> 00:51:02,276 SCIENTIFIC ADVANCEMENT WAS 1014 00:51:02,276 --> 00:51:05,746 INCONVENIENT AND CHALLENGING, 1015 00:51:05,746 --> 00:51:06,848 BUT IT WAS CRUCIAL FOR 1016 00:51:06,848 --> 00:51:10,685 SUSTAINING A LONG AND PRODUCTIVE 1017 00:51:10,685 --> 00:51:12,553 CAREER. 1018 00:51:12,553 --> 00:51:14,722 OUR CONTRIBUTION TRANSFORMED 1019 00:51:14,722 --> 00:51:18,793 WHAT WAS ONCE CONSIDERED A 1020 00:51:18,793 --> 00:51:20,194 OBSCURE PATHOGEN, THERE WAS JUST 1021 00:51:20,194 --> 00:51:21,996 300 CASES A YEAR, NEW CASES, 1022 00:51:21,996 --> 00:51:24,599 UNTIL AIDS CAME IN. 1023 00:51:24,599 --> 00:51:26,567 INTO A MODEL ORGANISM IN 1024 00:51:26,567 --> 00:51:30,338 BIOMEDICAL RESEARCH. 1025 00:51:30,338 --> 00:51:33,374 AS HIV EPIDEMIC HEIGHTENED THE 1026 00:51:33,374 --> 00:51:35,476 IMPORTANCE OF CRYPTO COCCOSIS. 1027 00:51:35,476 --> 00:51:37,678 I AM DEEPLY INDEBTED TO MY 1028 00:51:37,678 --> 00:51:42,049 MENTORS AND DEDICATED CO-WORKERS 1029 00:51:42,049 --> 00:51:43,551 AND SUPPORTIVE COLLEAGUES WHO 1030 00:51:43,551 --> 00:51:46,754 NUMBER MORE THAN 400 TODAY DATE. 1031 00:51:46,754 --> 00:51:48,122 THANK YOU. 1032 00:51:48,122 --> 00:51:58,299 [APPLAUSE] 1033 00:51:58,866 --> 00:52:00,301 >> LOOKS LIKE WE MIGHT HAVE A 1034 00:52:00,301 --> 00:52:03,070 QUESTION OVER HERE. 1035 00:52:03,070 --> 00:52:04,539 >> I HAVE A NAIVE QUESTION. 1036 00:52:04,539 --> 00:52:04,939 >> PLEASE. 1037 00:52:04,939 --> 00:52:06,140 >> HOW DOES THE MELANIN 1038 00:52:06,140 --> 00:52:07,675 CONTRIBUTE TO VIRULENCE? 1039 00:52:07,675 --> 00:52:08,442 IS THAT KNOWN? 1040 00:52:08,442 --> 00:52:09,043 >> EXCUSE ME? 1041 00:52:09,043 --> 00:52:10,411 >> HOW DOES THE MELANIN 1042 00:52:10,411 --> 00:52:11,412 CONTRIBUTE TO THE VIRULENCE? 1043 00:52:11,412 --> 00:52:13,381 >> OH. 1044 00:52:13,381 --> 00:52:14,348 MANY, MANY DIFFERENT WAYS. 1045 00:52:14,348 --> 00:52:18,085 I THINK THAT THE BEST WAY WILL 1046 00:52:18,085 --> 00:52:19,854 BE PETER WILLIAMS WILL ANSWER, 1047 00:52:19,854 --> 00:52:23,658 BUT IT PROTECTS THE CRYPTOCOCCUS 1048 00:52:23,658 --> 00:52:25,326 BECAUSE IT'S ANTIOXIDANT, AND 1049 00:52:25,326 --> 00:52:26,827 THEN OF COURSE IT'S ACCUMULATED 1050 00:52:26,827 --> 00:52:32,867 IN THE CELL WALL, SO ONCE -- THE 1051 00:52:32,867 --> 00:52:33,935 MEL NIEZED CELLS ARE MUCH MORE 1052 00:52:33,935 --> 00:52:34,735 DIFFICULT TO BE KILLED. 1053 00:52:34,735 --> 00:52:41,409 >> THANK YOU. 1054 00:52:41,409 --> 00:52:42,543 >> ACTUALLY I HAVE A QUESTION 1055 00:52:42,543 --> 00:52:43,377 FOR YOU. 1056 00:52:43,377 --> 00:52:46,514 YOU KNOW, I'M THINKING BACK ON 1057 00:52:46,514 --> 00:52:50,284 WHEN I WAS DOING CLINICAL WORK 1058 00:52:50,284 --> 00:52:52,486 AND MOST OF THE PEOPLE I SAW 1059 00:52:52,486 --> 00:52:58,059 WITH CRYPT CO CRYPTOCOCCAL MENIS 1060 00:52:58,059 --> 00:53:00,494 A NEUROLOGIST WERE PEOPLE WITH 1061 00:53:00,494 --> 00:53:02,230 DIABETES THAT WAS OUT OF 1062 00:53:02,230 --> 00:53:02,463 CONTROL. 1063 00:53:02,463 --> 00:53:02,730 >> RIGHT. 1064 00:53:02,730 --> 00:53:06,067 >> AND YET THE CRYPTOCOCCUS 1065 00:53:06,067 --> 00:53:10,238 APPARENTLY CAN LIVE IN LOW 1066 00:53:10,238 --> 00:53:12,540 GLUCOSE ENVIRONMENTS. 1067 00:53:12,540 --> 00:53:16,811 IS IT THE IMMUNE SYSTEM OF THE 1068 00:53:16,811 --> 00:53:19,547 UNCONTROLLED DIABETIC THAT IS 1069 00:53:19,547 --> 00:53:21,282 SOMETHING UNRELATED TO THE 1070 00:53:21,282 --> 00:53:22,984 GLUCOSE CONTENT OF THEIR BLOOD? 1071 00:53:22,984 --> 00:53:26,454 >> MAYBE I DIDN'T MENTION IT, 1072 00:53:26,454 --> 00:53:28,522 BUT THIS CRYPTOCOCCOSIS IS 1073 00:53:28,522 --> 00:53:30,224 PRIMARILY FOR IMMUNODEFICIENT 1074 00:53:30,224 --> 00:53:31,158 PATIENTS. 1075 00:53:31,158 --> 00:53:32,026 NUMBER ONE. 1076 00:53:32,026 --> 00:53:34,895 SO I'M SURE DIABETES PATIENTS 1077 00:53:34,895 --> 00:53:37,298 MUST BELONG THERE TOO. 1078 00:53:37,298 --> 00:53:40,501 BUT THE REASON WHY IT CAN THRIVE 1079 00:53:40,501 --> 00:53:42,436 IN THE BRAIN IS BECAUSE THIS 1080 00:53:42,436 --> 00:53:44,105 ORGANISM CAN THRIVE ON LOW 1081 00:53:44,105 --> 00:53:44,739 GLUCOSE MEDIA. 1082 00:53:44,739 --> 00:53:50,244 NOT THAT CRYPTO -- JUST LIKE THH 1083 00:53:50,244 --> 00:53:51,345 GLUCOSE BUT THEY CAN SURVIVE 1084 00:53:51,345 --> 00:53:53,047 VERY WELL IN LOW GLUCOSE. 1085 00:53:53,047 --> 00:53:53,648 >> THANK YOU. 1086 00:53:53,648 --> 00:53:55,082 THANK YOU VERY, VERY MUCH. 1087 00:53:55,082 --> 00:54:01,088 [APPLAUSE] 1088 00:54:01,088 --> 00:54:02,957 OUR NEXT SPEAKER IS DR. THOMAS 1089 00:54:02,957 --> 00:54:03,758 KUNKEL. 1090 00:54:03,758 --> 00:54:13,934 [APPLAUSE] 1091 00:55:06,821 --> 00:55:08,556 >> IT'S A GREAT PLEASURE TO BE 1092 00:55:08,556 --> 00:55:08,923 HERE. 1093 00:55:08,923 --> 00:55:15,262 I'VE NEVER HAD TO GIVE A TALK 1094 00:55:15,262 --> 00:55:16,697 THAT WAS DESCRIBED AS IT WAS TO 1095 00:55:16,697 --> 00:55:20,634 ME TO TRY TO COVER 42 YEARS OF 1096 00:55:20,634 --> 00:55:27,975 RESEARCH IN 30 MINUTES. 1097 00:55:27,975 --> 00:55:30,945 SO I'M GOING TO TRY DO THAT WITH 1098 00:55:30,945 --> 00:55:35,549 MINIMAL NEW DATA, BUT TO 1099 00:55:35,549 --> 00:55:36,851 DESCRIBE SOME OF THE THINGS THAT 1100 00:55:36,851 --> 00:55:39,186 HAVE REALLY STRUCK ME IN MY TIME 1101 00:55:39,186 --> 00:55:44,658 AS A SCIENTIST. 1102 00:55:44,658 --> 00:55:46,727 SO I WAS AS A CHILD A MEMBER OF 1103 00:55:46,727 --> 00:55:51,399 A LARGE FAMILY, 11 OF US, SIX 1104 00:55:51,399 --> 00:55:52,867 BROTHERS AND TWO SISTERS AND MY 1105 00:55:52,867 --> 00:55:54,635 PARENTS, AND WE GREW UP IN A 1106 00:55:54,635 --> 00:55:56,370 TINY LITTLE TOBACCO FARM IN THE 1107 00:55:56,370 --> 00:56:00,541 HILLS OF KENTUCKY. 1108 00:56:00,541 --> 00:56:02,076 AND THE FURTHEST THING FROM MY 1109 00:56:02,076 --> 00:56:03,411 MIND AS A CHILD WAS TO BECOME A 1110 00:56:03,411 --> 00:56:08,649 SCIENTIST. 1111 00:56:08,649 --> 00:56:10,418 AND WHAT LED TO THAT WAS THE 1112 00:56:10,418 --> 00:56:15,122 FACT THAT MY FATHER HAD A HEART 1113 00:56:15,122 --> 00:56:16,457 CONDITION AND PASSED AWAY AT THE 1114 00:56:16,457 --> 00:56:18,325 AGE OF 60, WHEN I WAS VERY 1115 00:56:18,325 --> 00:56:21,162 YOUNG, AND HAD TO -- AND WE HAD 1116 00:56:21,162 --> 00:56:25,232 TO MOVE INTO THE CITY, AND THAT 1117 00:56:25,232 --> 00:56:29,270 ACTUALLY BROKE MY HEART BECAUSE 1118 00:56:29,270 --> 00:56:30,805 UNTIL I WAS ABOUT 25 YEARS OLD, 1119 00:56:30,805 --> 00:56:32,339 I THOUGHT I WAS GOING TO BE A 1120 00:56:32,339 --> 00:56:33,474 FARMER, AND THAT WAS WHAT I 1121 00:56:33,474 --> 00:56:35,242 DEARLY LOVED TO DO. 1122 00:56:35,242 --> 00:56:38,078 BUT AT THE SAME TIME, I ENJOYED 1123 00:56:38,078 --> 00:56:41,916 OTHER THINGS, LIKE VERY MUCH 1124 00:56:41,916 --> 00:56:47,688 ENJOYED SCIENCE FICTION. 1125 00:56:47,688 --> 00:56:49,023 AND I WOULD READ BOOKS ABOUT 1126 00:56:49,023 --> 00:56:51,192 SCIENCE FICTION FROM THE TIME I 1127 00:56:51,192 --> 00:56:53,027 WAS ABOUT SEVEN YEARS OLD UNTIL 1128 00:56:53,027 --> 00:56:57,064 TODAY. 1129 00:56:57,064 --> 00:56:58,365 AND I STILL ENJOY THAT. 1130 00:56:58,365 --> 00:57:02,536 AND THAT'S ILLUSTRATED BY THIS 1131 00:57:02,536 --> 00:57:03,971 TO YOUR LEFT-HAND SIDE SIDE, THE 1132 00:57:03,971 --> 00:57:10,544 MIBGYTHEMILKY WAY AS IT MIGHT AR 1133 00:57:10,544 --> 00:57:11,312 FROM 10 BILLION LIGHT YEARS 1134 00:57:11,312 --> 00:57:13,280 AWAY, AND THAT'S 10 TO THE 23RD 1135 00:57:13,280 --> 00:57:15,249 METERS AND WE'RE ALL SCIENTISTS 1136 00:57:15,249 --> 00:57:16,584 AND KNOW THAT THAT IS A HUGE 1137 00:57:16,584 --> 00:57:17,117 DISTANCE. 1138 00:57:17,117 --> 00:57:19,987 AND ON THE RIGHT IS WHAT I DO, 1139 00:57:19,987 --> 00:57:22,490 WHICH IS TO STUDY THE FIDELITY 1140 00:57:22,490 --> 00:57:25,025 OF DNA REPLICATION AND IT'S 1141 00:57:25,025 --> 00:57:26,760 ILLUSTRATED THERE BY THE X-RAY 1142 00:57:26,760 --> 00:57:33,467 CRYSTAL STRU STRUCTURE OF A DNA 1143 00:57:33,467 --> 00:57:35,669 POLYMERASE, CORRECTLY PAIRING A 1144 00:57:35,669 --> 00:57:38,205 INCOMING OPPOSITE A T. 1145 00:57:38,205 --> 00:57:41,942 AND THE DISTANCE THERE IS 1146 00:57:41,942 --> 00:57:42,910 ABSOLUTELY HUGE BETWEEN THE LEFT 1147 00:57:42,910 --> 00:57:47,281 AND THE RIGHT. 1148 00:57:47,281 --> 00:57:50,751 IN IS THE EVOLUTION OF LIFE ON 1149 00:57:50,751 --> 00:57:53,220 EARTH AS PRESENTED TO ME BY MY 1150 00:57:53,220 --> 00:57:56,724 EXCELLENT COLLEAGUE, ROELLE 1151 00:57:56,724 --> 00:57:57,057 SHOPPER. 1152 00:57:57,057 --> 00:57:58,292 THE FORMATION OF THE MOON IS ON 1153 00:57:58,292 --> 00:58:01,095 THE LEFT AND THAT WAS 1154 00:58:01,095 --> 00:58:04,098 4.5 BILLION YEARS A AND THAT'S 1155 00:58:04,098 --> 00:58:04,965 ALMOST UNIMAGINABLE FOR US TO 1156 00:58:04,965 --> 00:58:06,834 THINK ABOUT WHAT A BILLION YEARS 1157 00:58:06,834 --> 00:58:10,337 IS. 1158 00:58:10,337 --> 00:58:11,739 AND ON THE RIGHT IS WHAT THE NIH 1159 00:58:11,739 --> 00:58:15,175 DOES, WHICH IS TO UNDERSTAND THE 1160 00:58:15,175 --> 00:58:15,843 IMPACT OF THE ENVIRONMENT ON 1161 00:58:15,843 --> 00:58:18,779 HUMAN HEALTH. 1162 00:58:18,779 --> 00:58:20,748 AND YOU CAN SEE OVER THERE, HOME 1163 00:58:20,748 --> 00:58:22,416 SAIPIANS HAVE ONLY BEEN AROUND 1164 00:58:22,416 --> 00:58:24,385 FOR A VERY, VERY, VERY TINY 1165 00:58:24,385 --> 00:58:28,322 SLICE OF TIME. 1166 00:58:28,322 --> 00:58:29,623 SO ONE OF THE THINGS THAT HAS 1167 00:58:29,623 --> 00:58:33,561 ALWAYS INTRIGUED ME IS HOW DID 1168 00:58:33,561 --> 00:58:37,398 LIFE EVOLVE ON EARTH OVER 4 1/2 1169 00:58:37,398 --> 00:58:42,303 BILLION YEARS? 1170 00:58:42,303 --> 00:58:45,039 SO TIME IS A VERY CRITICAL 1171 00:58:45,039 --> 00:58:45,639 DETERMINANT OF THE ANSWER TO 1172 00:58:45,639 --> 00:58:49,009 THAT QUESTION. 1173 00:58:49,009 --> 00:58:50,878 THIS IS ANOTHER IMAGE PROVIDED 1174 00:58:50,878 --> 00:58:52,746 TO ME BY ANOTHER GOOD COLLEAGUE 1175 00:58:52,746 --> 00:58:56,350 OF MINE AT THE NIEHS, 1176 00:58:56,350 --> 00:58:58,018 ENVIRONMENTAL HEALTH SCIENCES 1177 00:58:58,018 --> 00:58:58,285 INSTITUTE. 1178 00:58:58,285 --> 00:59:00,754 WE LIVE IN NORTH CAROLINA, SO I 1179 00:59:00,754 --> 00:59:03,591 DON'T GET TO WALK DOWN YOUR 1180 00:59:03,591 --> 00:59:04,491 BEAUTIFUL HALLWAYS HERE. 1181 00:59:04,491 --> 00:59:08,128 I WAS VERY IMPRESSED WITH ALL OF 1182 00:59:08,128 --> 00:59:09,530 THE STUFF THAT YOU GUYS PROBABLY 1183 00:59:09,530 --> 00:59:12,132 SEE EVERY DAY AND MAYBE DON'T 1184 00:59:12,132 --> 00:59:13,667 EVEN PAY ANY ATTENTION TO 1185 00:59:13,667 --> 00:59:19,173 ANYMORE, WHICH IS THE LIST OF 1186 00:59:19,173 --> 00:59:20,374 PREVIOUS COLLEAGUES OF YOURS WHO 1187 00:59:20,374 --> 00:59:23,544 WON PRIZES AND DISCOVERED 1188 00:59:23,544 --> 00:59:27,381 BIOLOGY AS WE KNOW IT TODAY, AND 1189 00:59:27,381 --> 00:59:28,716 I TOOK QUITE A WHILE COMING DOWN 1190 00:59:28,716 --> 00:59:31,018 THE HALLWAY HERE TO LIPSETT TO 1191 00:59:31,018 --> 00:59:32,987 READ SOME OF THAT STUFF LIKE 1192 00:59:32,987 --> 00:59:40,127 RIGHT OUTSIDE IS MARSHAL 1193 00:59:40,127 --> 00:59:42,630 NUREMBERG AND MARSHAL NUREMBERG 1194 00:59:42,630 --> 00:59:43,764 WAS THE MENTOR FOR ONE OF MY 1195 00:59:43,764 --> 00:59:45,466 BEST FRIENDS IN THE WORLD WHO 1196 00:59:45,466 --> 00:59:47,267 RECENTLY PASSED AWAY, SO I'M NOT 1197 00:59:47,267 --> 00:59:50,237 THAT FAR REMOVED BIOLOGICALLY 1198 00:59:50,237 --> 00:59:52,873 FROM THE TIME FRAME THAT MADE 1199 00:59:52,873 --> 00:59:55,309 NIH GREAT, AND THE BASIC 1200 00:59:55,309 --> 00:59:56,477 RESEARCH THAT DETERMINES LIFE ON 1201 00:59:56,477 --> 01:00:00,648 EARTH. 1202 01:00:00,648 --> 01:00:04,284 JEFF NUELER PROVIDED ME THIS, 1203 01:00:04,284 --> 01:00:08,122 HE'S AN NMR SPECTROSCOPIST. 1204 01:00:08,122 --> 01:00:09,990 THE TOOLS USED ON THE BOTTOM TO 1205 01:00:09,990 --> 01:00:14,161 DISCOVER PROTEIN DYNAMICS AND 1206 01:00:14,161 --> 01:00:16,230 MOLECULAR MOTIONS USING NUCLEAR 1207 01:00:16,230 --> 01:00:17,865 MAGNETIC RESONANCE, AND THE TIME 1208 01:00:17,865 --> 01:00:21,201 FRAME HERE IS VERY, VERY 1209 01:00:21,201 --> 01:00:22,670 DIFFERENT THAN THE 4 BILLION 1210 01:00:22,670 --> 01:00:24,972 YEARS OF EVOLUTION THAT I 1211 01:00:24,972 --> 01:00:25,239 DESCRIBED. 1212 01:00:25,239 --> 01:00:26,807 IN FACT, IF YOU WANT TO 1213 01:00:26,807 --> 01:00:28,942 UNDERSTAND REPLICATION FIDELITY, 1214 01:00:28,942 --> 01:00:31,278 THEN YOU NEED TO UNDERSTAND WHAT 1215 01:00:31,278 --> 01:00:32,946 GOES ON IN THE X-RAY CRYSTAL 1216 01:00:32,946 --> 01:00:34,815 STRUCTURE OF A DNA POLYMERASE 1217 01:00:34,815 --> 01:00:36,350 WHERE THE LENGTH OF A HYDROGEN 1218 01:00:36,350 --> 01:00:38,986 BOND BETWEEN AN A AND A T BASE 1219 01:00:38,986 --> 01:00:42,956 PAIR IS 2 ANGSTROMS, TO ROUGHLY 1220 01:00:42,956 --> 01:00:45,726 THE SIZE OF THE HUMAN BEING, AS 1221 01:00:45,726 --> 01:00:47,995 WE COULD ROUGHLY DESCRIBE AS 1222 01:00:47,995 --> 01:00:49,897 BEING 2 METERS TALL, AND IT 1223 01:00:49,897 --> 01:00:52,499 COVERS A TIME FRAME OF 10 TO THE 1224 01:00:52,499 --> 01:00:54,735 MINUS 14TH SECONDS TO GREATER 1225 01:00:54,735 --> 01:00:56,470 THAN 4 BILLION YEARS. 1226 01:00:56,470 --> 01:00:59,973 AND IN ORDER TO COVER THAT 1227 01:00:59,973 --> 01:01:02,376 IMMENSE AMOUNT OF TERRITORY TO 1228 01:01:02,376 --> 01:01:04,712 UNDERSTAND DNA REPLICATION 1229 01:01:04,712 --> 01:01:06,346 FIDELITY, WE NEED FOUR BASIC 1230 01:01:06,346 --> 01:01:07,548 TECHNOLOGIES, WHICH I DESCRIBE 1231 01:01:07,548 --> 01:01:09,450 AS STRUCTURAL BIOLOGY, 1232 01:01:09,450 --> 01:01:10,517 BIOCHEMISTRY, GENETICS, AND 1233 01:01:10,517 --> 01:01:15,355 GENOMICS. 1234 01:01:15,355 --> 01:01:18,192 IF YOU WANT TO TYPE THE HUMAN 1235 01:01:18,192 --> 01:01:22,062 GENOME, IT'S 6 BILLION 1236 01:01:22,062 --> 01:01:24,131 NUCLEOTIDES, AND IF YOU WERE 1237 01:01:24,131 --> 01:01:27,835 TYPING ON A WORD PROCESSOR, YOU 1238 01:01:27,835 --> 01:01:30,170 WOULD USE -- SPELL SELECTING THE 1239 01:01:30,170 --> 01:01:32,139 CORRECT BET LETTER OF THE 1240 01:01:32,139 --> 01:01:34,675 ALPHABET, YOU WOULD USE BACK 1241 01:01:34,675 --> 01:01:36,877 SPACE, AND YOU WOULD USE SPELL 1242 01:01:36,877 --> 01:01:38,846 CHECK TO ULTIMATELY GET AN 1243 01:01:38,846 --> 01:01:39,413 ERROR-FREE DOCUMENT. 1244 01:01:39,413 --> 01:01:40,914 NOW I HAD A SLIDE THAT I TOOK 1245 01:01:40,914 --> 01:01:44,351 OUT THAT WAS 500 LETTERS, AND I 1246 01:01:44,351 --> 01:01:45,719 HAD STUDENTS COME IN TO THE LAB 1247 01:01:45,719 --> 01:01:49,289 OVER THE YEARS WHO USED ONE, TWO 1248 01:01:49,289 --> 01:01:52,159 OR THREE OF THOSE METHODS TO 1249 01:01:52,159 --> 01:01:54,228 TYPE THAT 500 LETTERS, AND 1250 01:01:54,228 --> 01:01:57,498 THERE'S THEIR ERROR RATES. 1251 01:01:57,498 --> 01:02:00,734 ABOUT 20 MISTAKES FOR EVERY 500 1252 01:02:00,734 --> 01:02:02,803 NUCLEOTIDES IF ALL YOU USE IS 1253 01:02:02,803 --> 01:02:03,237 THE FIRST STEP. 1254 01:02:03,237 --> 01:02:05,239 IF YOU ADD THE SECOND STEP, YOU 1255 01:02:05,239 --> 01:02:07,875 IMPROVE FIDELITY, AND IF YOU ADD 1256 01:02:07,875 --> 01:02:11,378 SPELL CHECK, YOU GET AN 1257 01:02:11,378 --> 01:02:12,579 UNBLEMISHED DOCUMENT. 1258 01:02:12,579 --> 01:02:14,148 SAME HOLDS FOR REPLICATION 1259 01:02:14,148 --> 01:02:16,884 FIDELITY. 1260 01:02:16,884 --> 01:02:19,286 WHERE THE DNA POLYMERASE THAT 1261 01:02:19,286 --> 01:02:21,355 SELECTS THE CORRECT NUCLEOTIDE 1262 01:02:21,355 --> 01:02:23,123 HAS A CERTAIN SELECTIVITY FOR A 1263 01:02:23,123 --> 01:02:25,759 CORRECT OVER AN INCORRECT 1264 01:02:25,759 --> 01:02:26,860 NUCLEOTIDE. 1265 01:02:26,860 --> 01:02:29,496 PROOFREADING, THAT IS, THE 1266 01:02:29,496 --> 01:02:31,465 EXO -- REMOVAL OF A MISMATCH IS 1267 01:02:31,465 --> 01:02:33,467 ROUGHLY EQUIVALENT TO BACKSPACE, 1268 01:02:33,467 --> 01:02:36,436 AND SPELLCHECK COULD BE ROUGHLY 1269 01:02:36,436 --> 01:02:37,838 EQUIVALENT TO TWO MAJOR REPAIR 1270 01:02:37,838 --> 01:02:39,807 PROCESSES THAT I WOULD LIKE TO 1271 01:02:39,807 --> 01:02:41,341 BRIEFLY DESCRIBE IN THE NEXT FEW 1272 01:02:41,341 --> 01:02:41,909 MINUTES. 1273 01:02:41,909 --> 01:02:43,577 ONE IS DNA MISMATCH REPAIR AND 1274 01:02:43,577 --> 01:02:48,182 THE OTHER IS RIBONUCLEOTIDE 1275 01:02:48,182 --> 01:02:49,383 EXCISION REPAIR. 1276 01:02:49,383 --> 01:02:51,285 THE WORLD'S EXPERT ON THAT 1277 01:02:51,285 --> 01:02:52,586 SUBJECT IS SOMEWHERE IN THIS 1278 01:02:52,586 --> 01:02:55,222 AUDIENCE LISTENING TO ME. 1279 01:02:55,222 --> 01:02:57,191 THAT'S BOB KROUCH. 1280 01:02:57,191 --> 01:02:59,393 AND WHAT WE HAVE INTN 1281 01:02:59,393 --> 01:03:00,928 INVESTIGATING FOR THE 42 YEARS 1282 01:03:00,928 --> 01:03:03,030 THAT I'VE BEEN AT THE NIEHS WAS 1283 01:03:03,030 --> 01:03:05,766 TO TRY TO UNDERSTAND HOW 1284 01:03:05,766 --> 01:03:07,534 REPLICATION FIDELITY CAN BE WHAT 1285 01:03:07,534 --> 01:03:10,370 I CONSIDER ONE OF THE MOST 1286 01:03:10,370 --> 01:03:12,773 AMAZINGLY ACCURATE BIOLOGICAL 1287 01:03:12,773 --> 01:03:17,311 PROCESSES THAT OCCUR, AND I'M 1288 01:03:17,311 --> 01:03:18,478 GOING TO GIVE YOU A LITTLE BIT 1289 01:03:18,478 --> 01:03:20,948 ABOUT THAT, AS A PERSON WHO WAS 1290 01:03:20,948 --> 01:03:22,916 ORIGINALLY TRAINED AS A 1291 01:03:22,916 --> 01:03:24,785 BIOCHEMIST AND PICK UNDER 1292 01:03:24,785 --> 01:03:27,621 STRUCTURAL BIOLOGY, GENETICS AND 1293 01:03:27,621 --> 01:03:33,660 GENOMICS AS TIME WENT BY. 1294 01:03:33,660 --> 01:03:35,195 SO THIS IS WHAT WE BELIEVE 1295 01:03:35,195 --> 01:03:37,831 SUMMARIZES WHAT IT TAKES TO 1296 01:03:37,831 --> 01:03:40,367 REPLICATE THE 6 BILLION 1297 01:03:40,367 --> 01:03:42,569 NUCLEOTIDES OF DNA IN THE 1298 01:03:42,569 --> 01:03:48,575 NUCLEAR GENOME OF A HUMAN BEING. 1299 01:03:48,575 --> 01:03:49,843 ONE, TWO, THREE AND FOUR ARE THE 1300 01:03:49,843 --> 01:03:51,378 PROCESSES THAT WE HAVE BEEN 1301 01:03:51,378 --> 01:03:53,447 VIGOROUSLY INVESTIGATING FOR THE 1302 01:03:53,447 --> 01:03:55,449 42 YEARS THAT I'VE BEEN AT THE 1303 01:03:55,449 --> 01:03:57,618 NIEHS, AND MOST OF THAT WAS ON 1304 01:03:57,618 --> 01:03:59,419 THE SELECTIVITY OF THE DNA POE 1305 01:03:59,419 --> 01:04:02,589 LPOLYMERASES THEMSELVES, NUMBER 1306 01:04:02,589 --> 01:04:04,324 TWO, IN WHICH YOU HAVE A CORRECT 1307 01:04:04,324 --> 01:04:08,295 PROPERLY ALIGNED DNTP INCOMING 1308 01:04:08,295 --> 01:04:10,197 OPPOSITE A CORRECT TEMPLATE 1309 01:04:10,197 --> 01:04:12,599 BASE, AND YOU ALSO HAVE THE 1310 01:04:12,599 --> 01:04:16,670 CORRECT SUGAR MOIETY, 1311 01:04:16,670 --> 01:04:18,438 DEOXYRIBONUCLEOTIDES, INSTEAD OF 1312 01:04:18,438 --> 01:04:19,539 RIBONUCLEOTIDES. 1313 01:04:19,539 --> 01:04:21,942 BUT OVER TIME, WE HAVE 1314 01:04:21,942 --> 01:04:23,510 INVESTIGATED ALL THOSE 1315 01:04:23,510 --> 01:04:25,045 PROCESSES, AND I DON'T HAVE TIME 1316 01:04:25,045 --> 01:04:27,648 TO DESCRIBE TO YOU EVERYTHING 1317 01:04:27,648 --> 01:04:28,649 THAT WE'VE LEARNED ABOUT THOSE, 1318 01:04:28,649 --> 01:04:31,418 SO I PICKED A COUPLE OF SELECTED 1319 01:04:31,418 --> 01:04:33,420 SUBJECTS THAT HAVE HAD AN IMPACT 1320 01:04:33,420 --> 01:04:35,789 ON HOW WE THINK ABOUT 1321 01:04:35,789 --> 01:04:37,925 REPLICATION FIDELITY, AND BY 1322 01:04:37,925 --> 01:04:39,593 THAT, I MEAN HOW THE WORLD 1323 01:04:39,593 --> 01:04:42,562 THINKS ABOUT IT BASED OP WH ON S 1324 01:04:42,562 --> 01:04:44,097 NOW IN MOLECULAR BAYOLOGY 1325 01:04:44,097 --> 01:04:45,966 TEXTBOOKS, PARTLY BASED ON WHAT 1326 01:04:45,966 --> 01:04:49,369 MY LAB HAS CONTRIBUTED TO THIS 1327 01:04:49,369 --> 01:04:54,041 AREA OF INVESTIGATION. 1328 01:04:54,041 --> 01:04:56,510 AND THE REASON WE'RE INTERESTED 1329 01:04:56,510 --> 01:04:58,612 IS THE RESPONSIBLE MECHANISMS 1330 01:04:58,612 --> 01:05:01,281 THAT DETERMINE REPLICATION 1331 01:05:01,281 --> 01:05:02,616 FIDELITY, THEIR CONNECTIONS TO 1332 01:05:02,616 --> 01:05:06,219 HUMAN HEALTH WHICH IS SIMILAR TO 1333 01:05:06,219 --> 01:05:07,754 WHAT THE NIH DOES AS AN 1334 01:05:07,754 --> 01:05:09,556 INSTITUTION OR A SET OF 1335 01:05:09,556 --> 01:05:10,958 INSTITUTIONS, AND THE 1336 01:05:10,958 --> 01:05:11,825 EVOLUTIONARY CONSERVATION OF 1337 01:05:11,825 --> 01:05:14,261 THOSE PROCESSES, WHICH IS MORE 1338 01:05:14,261 --> 01:05:16,163 OF A PERSONAL INTEREST ON MY 1339 01:05:16,163 --> 01:05:17,798 PART, BUT NONETHELESS, IMPORTANT 1340 01:05:17,798 --> 01:05:19,099 TO HUMAN HEALTH. 1341 01:05:19,099 --> 01:05:21,234 AND THE PROCESSES THAT WE THINK 1342 01:05:21,234 --> 01:05:23,537 ABOUT ARE THE ELONGATION OF DNA 1343 01:05:23,537 --> 01:05:27,674 ON AN OPEN TEMPLATE WITHOUT A 1344 01:05:27,674 --> 01:05:28,976 DOWNSTREAM DOUBLE STRANDED 1345 01:05:28,976 --> 01:05:29,810 NATURE. 1346 01:05:29,810 --> 01:05:33,847 SO IT'S ADDING NUCLEOTIDES TO A 1347 01:05:33,847 --> 01:05:37,150 TEMPLATE PRIMER FOR 6 BILLION 1348 01:05:37,150 --> 01:05:38,151 NUCLEOTIDES AND DOING IT WITHOUT 1349 01:05:38,151 --> 01:05:42,522 MAKING A MISTAKE. 1350 01:05:42,522 --> 01:05:45,692 AND THEN THERE'S OKAZAKI 1351 01:05:45,692 --> 01:05:46,860 FRAGMENT MATURATION WHICH IS THE 1352 01:05:46,860 --> 01:05:48,395 PROCESS WHICH ON THE LAGGING 1353 01:05:48,395 --> 01:05:52,099 STRAND OF REPLICATION ON WHICH 1354 01:05:52,099 --> 01:05:53,133 OKAZAKI FRAGMENTS ARE MADE, YOU 1355 01:05:53,133 --> 01:05:54,868 HAVE TO GET RID OF THE RNA 1356 01:05:54,868 --> 01:05:56,670 PRIMER THAT WAS USED TO INITIATE 1357 01:05:56,670 --> 01:05:58,872 THOSE OKAZAKI FRAGMENTS AND YOU 1358 01:05:58,872 --> 01:06:00,741 DO THAT BY A STRAND DISPLACEMENT 1359 01:06:00,741 --> 01:06:01,008 REACTION. 1360 01:06:01,008 --> 01:06:03,010 AND THAT OCCURS ABOUT 25 MILLION 1361 01:06:03,010 --> 01:06:04,378 TIMES PER REPLICATION OF THE 1362 01:06:04,378 --> 01:06:05,145 HUMAN GENOME. 1363 01:06:05,145 --> 01:06:07,214 AND THEN THE POST REPLICATION 1364 01:06:07,214 --> 01:06:09,149 REPAIR PROCESSES, THERE ARE TONS 1365 01:06:09,149 --> 01:06:11,752 OF REPLICATION -- OR REPAIR 1366 01:06:11,752 --> 01:06:13,820 PROCESSES THAT OCCUR IN CELLS. 1367 01:06:13,820 --> 01:06:17,057 THEY'RE DONE BY THE 17 NORMAL 1368 01:06:17,057 --> 01:06:19,026 HUMAN DNA POLYMERASES, WHICH MY 1369 01:06:19,026 --> 01:06:20,327 LAB INVESTIGATE WOULD FOR MORE 1370 01:06:20,327 --> 01:06:26,767 THAN 25 YEARS. 1371 01:06:26,767 --> 01:06:29,002 AND THEY OCCUR BY MISMATCH OF 1372 01:06:29,002 --> 01:06:30,604 THOSE MANY DNA REPAIR PROCESSES, 1373 01:06:30,604 --> 01:06:32,072 TWO OF THEM ARE PARTICULARLY 1374 01:06:32,072 --> 01:06:33,740 IMPORTANT TO THE FIDELITY OF 1375 01:06:33,740 --> 01:06:36,043 REPLICATION OF THE HUMAN NUCLEAR 1376 01:06:36,043 --> 01:06:36,276 GENOME. 1377 01:06:36,276 --> 01:06:39,446 SO THESE ARE THE 17 DNA 1378 01:06:39,446 --> 01:06:40,313 POLYMERASES THAT WE KNOW ABOUT 1379 01:06:40,313 --> 01:06:41,048 TO DATE. 1380 01:06:41,048 --> 01:06:43,350 IN MY LAB OVER TIME, WE'VE 1381 01:06:43,350 --> 01:06:46,086 INVESTIGATED THE FIDELITY WITH 1382 01:06:46,086 --> 01:06:48,121 WHICH THEY CAN REPLICATE DNA 1383 01:06:48,121 --> 01:06:50,157 WITHOUT MAKING A MISTAKE. 1384 01:06:50,157 --> 01:06:52,759 AND I'M GOING TO LEAVE ABOUT 1385 01:06:52,759 --> 01:06:54,961 HALF OF THAT WORK BEHIND AND 1386 01:06:54,961 --> 01:06:57,431 FOCUS ON ONE ASPECT, THE 1387 01:06:57,431 --> 01:07:00,167 FIDELITY OF NUCLEAR DNA 1388 01:07:00,167 --> 01:07:01,368 REPLICATION, AND OUR CURRENT 1389 01:07:01,368 --> 01:07:05,072 WORK IS PRIMARILY DONE IN 1390 01:07:05,072 --> 01:07:09,376 BUDDING YEAST AS A LEGITIMATE 1391 01:07:09,376 --> 01:07:10,911 MOLECULAR MODEL FOR WHAT GOES ON 1392 01:07:10,911 --> 01:07:12,946 IN HUMANS. 1393 01:07:12,946 --> 01:07:16,049 SO THESE ARE THE THREE NORMAL 1394 01:07:16,049 --> 01:07:18,919 REPLICATIONS THAT REPLICATE THE 1395 01:07:18,919 --> 01:07:21,688 6 BILLION NUCLEOTIDES. 1396 01:07:21,688 --> 01:07:23,123 THEY'RE POLYMERASES ALPHA, DELTA 1397 01:07:23,123 --> 01:07:26,159 AND EPSILON OR 1, 2 AND 3, AND 1398 01:07:26,159 --> 01:07:27,294 THIS SLIDE IS JUST TO SHOW YOU 1399 01:07:27,294 --> 01:07:28,829 THAT THEY'RE HEAVILY CONSERVED 1400 01:07:28,829 --> 01:07:32,132 IN MAMMALS AND IN OTHER 1401 01:07:32,132 --> 01:07:34,901 EUKARYOTES INCLUDING THE TWO 1402 01:07:34,901 --> 01:07:36,303 YEAST ORGANISMS THAT WE HAVE 1403 01:07:36,303 --> 01:07:45,812 SOME INFORMATION ON S.CEREVIIAE 1404 01:07:45,812 --> 01:07:46,913 AND POMBE. 1405 01:07:46,913 --> 01:07:48,748 AT THE VERY BOTTOM, YOU'LL SEE 1406 01:07:48,748 --> 01:07:50,617 THEY HAVE A FIDELITY OF DNA 1407 01:07:50,617 --> 01:07:52,285 REPLICATION DONE BY OUR 1408 01:07:52,285 --> 01:07:53,120 BIOCHEMICAL MEASUREMENTS OVER 1409 01:07:53,120 --> 01:07:56,790 THE YEARS THAT ARE DIFFERENT, 1410 01:07:56,790 --> 01:07:59,292 POL ALPHA LACKS A 1411 01:07:59,292 --> 01:08:00,560 PROOFREADING -- AND ITS ACCURACY 1412 01:08:00,560 --> 01:08:02,696 IS ABOUT ONE MISTAKE FOR EVERY 1413 01:08:02,696 --> 01:08:11,671 2,000 NUCLEOTIDES PRELIMINARY -E 1414 01:08:11,671 --> 01:08:13,273 CURRENT WORK SUGGESTS THAT ALPHA 1415 01:08:13,273 --> 01:08:17,444 ONLY INITIATES SHORT CHAINS BY 1416 01:08:17,444 --> 01:08:19,079 LAYING DOWN AN RNA PRIMER AND 1417 01:08:19,079 --> 01:08:21,748 THEN EXTENDING IT BY A FEW MORE 1418 01:08:21,748 --> 01:08:23,950 NUCLEOTIDES WITH DNA, WHEREAS 1419 01:08:23,950 --> 01:08:25,585 DELTA AND EPSILON DO THE 1420 01:08:25,585 --> 01:08:29,789 MAJORITY OF LEADING AND -- 1421 01:08:29,789 --> 01:08:30,323 STRAND REPLICATION. 1422 01:08:30,323 --> 01:08:32,959 SO HOW DO WE KNOW ABOUT THE 1423 01:08:32,959 --> 01:08:34,661 FIDELITY OF LEADING AND LAGGING 1424 01:08:34,661 --> 01:08:35,529 STRAND REPLICATION? 1425 01:08:35,529 --> 01:08:36,530 WELL, HERE'S ONE WAY TO ADDRESS 1426 01:08:36,530 --> 01:08:37,164 THAT QUESTION. 1427 01:08:37,164 --> 01:08:41,902 WE CAN USE MUTATOR ALLELES OF 1428 01:08:41,902 --> 01:08:45,872 THOSE THREE REPLICATIONS WHICH 1429 01:08:45,872 --> 01:08:47,607 HAVE AN ACTIVE SITE POINT 1430 01:08:47,607 --> 01:08:49,709 MUTATION IN WHAT'S CALLED 1431 01:08:49,709 --> 01:08:52,913 LEUCINE 612 IN THE X-RAY CRYSTAL 1432 01:08:52,913 --> 01:08:57,350 STRUCTURE OF YEAST POL DELTA. 1433 01:08:57,350 --> 01:09:01,755 AND THAT -- WE CAN MAKE MUTANT 1434 01:09:01,755 --> 01:09:04,758 IN WHICH WE CHANGE THAT 1435 01:09:04,758 --> 01:09:06,493 LEUCINE -- OR ITS HOMOLOGUE IN 1436 01:09:06,493 --> 01:09:10,096 THE B FAMILY REPLICASE FAMILY IN 1437 01:09:10,096 --> 01:09:11,998 POL ALPHA IS AN EQUIVALENT 1438 01:09:11,998 --> 01:09:14,534 LEUCINE THAT WE CHANGED FROM ME 1439 01:09:14,534 --> 01:09:18,471 THOI NEEN OR IN EPSILON, THAT WE 1440 01:09:18,471 --> 01:09:20,473 CHANGED TO GLYCINE, AND WHAT WE 1441 01:09:20,473 --> 01:09:24,077 KNOW FROM YEARS OF WORK ON THOSE 1442 01:09:24,077 --> 01:09:25,845 PROTEINS IS THAT THEY HAVE 1443 01:09:25,845 --> 01:09:26,179 PREFERENCES. 1444 01:09:26,179 --> 01:09:31,051 YOU CAN MAKE 12 POSSIBLE 1445 01:09:31,051 --> 01:09:33,987 MISMATCHES IN ORDER TO GET BASE 1446 01:09:33,987 --> 01:09:35,088 SUBSTITUTION ERRORS, AND WE KNOW 1447 01:09:35,088 --> 01:09:37,290 THAT THEY ARE SELECTIVE IN 1448 01:09:37,290 --> 01:09:39,960 MAKING ONE PARTICULAR MISMATCH 1449 01:09:39,960 --> 01:09:41,494 OVER ANOTHER IN ORDER TO EXPLAIN 1450 01:09:41,494 --> 01:09:42,963 A BASE SUBSTITUTION MUTATION 1451 01:09:42,963 --> 01:09:45,165 THAT OCCURS IN A CELL. 1452 01:09:45,165 --> 01:09:49,536 SO FOR EXAMPLE, POL DELTA L TO M 1453 01:09:49,536 --> 01:09:52,472 HAS A 28 FOLD HIGHER PROBABILITY 1454 01:09:52,472 --> 01:09:54,941 OF INSERTING G OPPOSITE T THAN 1455 01:09:54,941 --> 01:09:58,311 IT HAS OF INSERTING C OPPOSITE 1456 01:09:58,311 --> 01:10:00,180 A, WHEN THE COMPLEMENTARY 1457 01:10:00,180 --> 01:10:04,050 STRAPPED IS STRANDIS BEING REPL. 1458 01:10:04,050 --> 01:10:05,585 SO WE CAN USE THAT KIND OF 1459 01:10:05,585 --> 01:10:06,886 REPLICATION FIDELITY ASYMMETRY 1460 01:10:06,886 --> 01:10:10,890 FOR EACH OF THESE MUTATOR 1461 01:10:10,890 --> 01:10:12,525 POLYMERASES AND ASK WHAT HAPPENS 1462 01:10:12,525 --> 01:10:15,095 WHEN YOU SEQUENCE A HUMAN 1463 01:10:15,095 --> 01:10:16,830 GENOME -- OR A YEAST GENOME 1464 01:10:16,830 --> 01:10:22,068 THAT'S BEEN THROW 30 PASSAGES ON 1465 01:10:22,068 --> 01:10:23,770 A PETRI DISH, WHICH IS 1466 01:10:23,770 --> 01:10:25,739 EQUIVALENT TO ABOUT 900 CELLULAR 1467 01:10:25,739 --> 01:10:28,141 GENERATIONS OR 900 REPLICATION 1468 01:10:28,141 --> 01:10:29,209 CYCLES. 1469 01:10:29,209 --> 01:10:30,710 AND WHAT YOU REALIZE WHEN YOU 1470 01:10:30,710 --> 01:10:34,648 SEQUENCE THE WHOLE GENOME OF A 1471 01:10:34,648 --> 01:10:40,120 POL DELTA L TO M MUTATOR ALLELE, 1472 01:10:40,120 --> 01:10:41,121 IS THAT THE KINDS OF MUTATIONS 1473 01:10:41,121 --> 01:10:43,723 THAT APPEAR IN THE CELL DEPEND 1474 01:10:43,723 --> 01:10:47,127 ON WHERE THE MUTATION IS AND 1475 01:10:47,127 --> 01:10:48,695 WHICH OF THE REPLICASES IS DOING 1476 01:10:48,695 --> 01:10:52,866 THE REPLICATION, SO IN THIS 1477 01:10:52,866 --> 01:10:56,903 EXAMPLE, POL DELTA MAKES ERRORS 1478 01:10:56,903 --> 01:10:59,372 ON THE LEFT-HAND SIDE THAT ARE 1479 01:10:59,372 --> 01:11:02,876 CONSISTENT WITH G OPPOSITE T, 1480 01:11:02,876 --> 01:11:04,744 AND ON THE RIGHT-HAND SIDE, WITH 1481 01:11:04,744 --> 01:11:06,846 ERRORS THAT ARE CONSISTENT WITH 1482 01:11:06,846 --> 01:11:08,281 G OPPOSITE T ON THE OPPOSITE 1483 01:11:08,281 --> 01:11:08,782 STRAND. 1484 01:11:08,782 --> 01:11:11,451 YOU CAN USE THIS KIND OF LOGIC 1485 01:11:11,451 --> 01:11:17,090 FOR ALL THREE REPLICASES AND ALL 1486 01:11:17,090 --> 01:11:19,192 12 BASE SPACED MISMATCHES AND 1487 01:11:19,192 --> 01:11:20,727 ARRIVE AT A MODEL THAT SUGGESTS 1488 01:11:20,727 --> 01:11:29,502 THAT PO POL ALPHA IS INITIATING- 1489 01:11:29,502 --> 01:11:32,138 CONTINUES TO DO THE BULK OF THE 1490 01:11:32,138 --> 01:11:34,274 FRAGMENTS AND POL EPSILON IS THE 1491 01:11:34,274 --> 01:11:37,110 MAJOR LEADING STRAND REPLICASE. 1492 01:11:37,110 --> 01:11:39,412 THAT'S A QUESTION THAT HAS 1493 01:11:39,412 --> 01:11:41,114 POSED -- HAS BEEN POSED FOR 1494 01:11:41,114 --> 01:11:44,951 YEARS, EVER SINCE WATSON AND 1495 01:11:44,951 --> 01:11:47,687 CRAIG DESCRIBED THE ASYMMETRIC 1496 01:11:47,687 --> 01:11:49,489 CONFIGURATION OF DNA, AND THE 1497 01:11:49,489 --> 01:11:52,525 DATA THAT WE GENERATED ON THESE 1498 01:11:52,525 --> 01:11:54,627 MUTATOR ALLELES WAS CONSISTENT 1499 01:11:54,627 --> 01:11:59,265 WITH THAT MODEL ON THE LOWER 1500 01:11:59,265 --> 01:11:59,632 RIGHT-HAND SIDE. 1501 01:11:59,632 --> 01:12:06,573 AND THAT'S NOW, I THINK, YOU 1502 01:12:06,573 --> 01:12:07,674 NEVER PROVE ANYTHING IN SCIENCE, 1503 01:12:07,674 --> 01:12:09,709 YOU ONLY TEST HYPOTHESES AND TRY 1504 01:12:09,709 --> 01:12:11,111 TO DISPROVE THEM, BUT IF YOU 1505 01:12:11,111 --> 01:12:13,980 CAN'T DISPROVE THEM WITH ANY 1506 01:12:13,980 --> 01:12:16,516 OBSERVATIONS YOU MAKE, THEN YOU 1507 01:12:16,516 --> 01:12:18,184 PUT FORWARD THE HYPOTHESIS THAT 1508 01:12:18,184 --> 01:12:20,153 YOU BELIEVE IS TRUE, AND I 1509 01:12:20,153 --> 01:12:24,958 BELIEVE THAT THAT MODEL DEFINES 1510 01:12:24,958 --> 01:12:27,293 THE ROLES OF THE THREE MAJOR 1511 01:12:27,293 --> 01:12:29,162 REPLICASES IN REPLICATING THE 1512 01:12:29,162 --> 01:12:30,230 LEADING AND THE LAGGING 1513 01:12:30,230 --> 01:12:31,464 STRAPPED. 1514 01:12:31,464 --> 01:12:33,500 AND THAT'S ONE OF THE 1515 01:12:33,500 --> 01:12:35,235 OBSERVATIONS THAT LED TO MY 1516 01:12:35,235 --> 01:12:36,836 ELECTION TO THE NATIONAL ACADEMY 1517 01:12:36,836 --> 01:12:39,539 OF SCIENCES IN MAY. 1518 01:12:39,539 --> 01:12:46,246 THAT'S NOT NEARLY ENOUGH. 1519 01:12:46,246 --> 01:12:48,381 SO WE ALSO, IN 2010, AND THIS IS 1520 01:12:48,381 --> 01:12:49,582 A STATEMENT ADDRESSED TO ANY OF 1521 01:12:49,582 --> 01:12:50,884 THE STUDENTS IN THE AUDIENCE, OR 1522 01:12:50,884 --> 01:12:52,552 THE YOUNGER STUDENTS, I'M STILL 1523 01:12:52,552 --> 01:12:58,558 A STUDENT, BUT -- PEOPLE TRYING 1524 01:12:58,558 --> 01:12:59,793 TO FIGURE OUT WHAT THEY WANT TO 1525 01:12:59,793 --> 01:13:02,228 BE WHEN THEY GROW UP, THEY MAY 1526 01:13:02,228 --> 01:13:04,697 WANT TO THINK ABOUT THIS. 1527 01:13:04,697 --> 01:13:05,498 RIBONUCLEOTIDES ARE NOT NORMALLY 1528 01:13:05,498 --> 01:13:07,200 CONSIDERED PART OF DNA BUT I GOT 1529 01:13:07,200 --> 01:13:08,601 A STUDENT IN THE LAB IN 2009 WHO 1530 01:13:08,601 --> 01:13:11,738 WORKED WITH MY POSTDOC AND FOUND 1531 01:13:11,738 --> 01:13:14,707 THAT RIBONUCLEOTIDES, THE WRONG 1532 01:13:14,707 --> 01:13:16,609 SUGAR, NOT THE WRONG BASE UP 1533 01:13:16,609 --> 01:13:19,813 THERE IN GREEN, ARE FREQUENTLY 1534 01:13:19,813 --> 01:13:21,114 INCORPORATED INTO DNA. 1535 01:13:21,114 --> 01:13:24,818 IN FACT, FOR YEARS, PEOPLE HAD 1536 01:13:24,818 --> 01:13:30,457 THOUGHT THAT DNA POLYMERASES 1537 01:13:30,457 --> 01:13:31,224 INCORPORATE -- TRIPHOSPHATES BUT 1538 01:13:31,224 --> 01:13:33,626 THEY'RE VERY, VERY POOR AT 1539 01:13:33,626 --> 01:13:36,129 INCORPORATING RIBONUCLEOTIDES. 1540 01:13:36,129 --> 01:13:38,064 BASED ON X-RAY CRYSTAL 1541 01:13:38,064 --> 01:13:38,832 STRUCTURE, YOU COULD IMAGINE 1542 01:13:38,832 --> 01:13:40,633 WERE THAT'S SEW BASED ON 1543 01:13:40,633 --> 01:13:43,403 CALCULATING THE DNTP AND THE 1544 01:13:43,403 --> 01:13:44,704 RNTP CONCENTRATIONS IN YEAST 1545 01:13:44,704 --> 01:13:48,608 CELLS BY OUR COLLEAGUE ANDREI 1546 01:13:48,608 --> 01:13:52,312 CHABES, WE DISCOVERED THAT DNTP 1547 01:13:52,312 --> 01:13:54,447 CONCENTRATIONS ARE MUCH, MUCH 1548 01:13:54,447 --> 01:13:58,751 LOWER THAN ARE RIBONUCLEOTIDE 1549 01:13:58,751 --> 01:13:59,486 INCORPORATIONS CONSISTENT WITH 1550 01:13:59,486 --> 01:14:02,622 MUCH MORE FREQUENT OR MUCH MORE 1551 01:14:02,622 --> 01:14:03,923 ABUNDANT RIBONUCLEOTIDE 1552 01:14:03,923 --> 01:14:07,126 SYNTHESIS TO MAKE THE REST OF 1553 01:14:07,126 --> 01:14:12,198 THE GENETIC CODE WORK THAN ARE 1554 01:14:12,198 --> 01:14:14,400 DEOXIES, BUT DNA POLYMERASES USE 1555 01:14:14,400 --> 01:14:23,276 MUCH LOWER DNTP CONCENTRATIONS. 1556 01:14:23,276 --> 01:14:24,844 THIS CAN BE PUT INTO THE FIELD 1557 01:14:24,844 --> 01:14:26,613 OF DNA REPAIR BY THE FOLLOWING. 1558 01:14:26,613 --> 01:14:28,281 THIS IS A SLIDE TAKEN FROM -- 1559 01:14:28,281 --> 01:14:31,050 GIVEN TO ME BY THOMAS LINDELL 1560 01:14:31,050 --> 01:14:33,686 ABOUT 20 YEARS AGO DESCRIBING -- 1561 01:14:33,686 --> 01:14:34,888 THOMAS WON THE NOBEL PRIZE FOR 1562 01:14:34,888 --> 01:14:37,790 HIS WORK ON BASE EXCISION 1563 01:14:37,790 --> 01:14:38,892 REPAIR, BUT HE WAS ALSO A VERY 1564 01:14:38,892 --> 01:14:43,429 STRONG POE PO PROPONENT OF DNA E 1565 01:14:43,429 --> 01:14:45,198 AND HOW IT AFFECTS REPAIR 1566 01:14:45,198 --> 01:14:46,533 PROCESSES, REPLICATION FIDELITY, 1567 01:14:46,533 --> 01:14:48,968 AND THIS IS A LIST OF THE 1568 01:14:48,968 --> 01:14:50,837 LESIONS IN DNA THAT HAD BEEN 1569 01:14:50,837 --> 01:14:54,607 STUDIED FOR ABOUT 50 YEARS AT 1570 01:14:54,607 --> 01:14:54,908 THAT POINT. 1571 01:14:54,908 --> 01:14:56,476 AND HOW FREQUENTLY THEY OCCUR IN 1572 01:14:56,476 --> 01:14:59,212 A HUMAN CELL IN THE THIRD -- OR 1573 01:14:59,212 --> 01:15:02,682 IN THE SECOND FROM THE RIGHT 1574 01:15:02,682 --> 01:15:03,082 COLUMN. 1575 01:15:03,082 --> 01:15:04,651 SO THAT'S THE NUMBER OF ABASIC 1576 01:15:04,651 --> 01:15:06,052 SITES, WHICH IS THE MOST 1577 01:15:06,052 --> 01:15:07,687 ABUNDANT LESION PRODUCED IN A 1578 01:15:07,687 --> 01:15:09,188 HUMAN CELL, AT LEAST AS FAR AS 1579 01:15:09,188 --> 01:15:11,691 WE KNEW 20 YEARS AGO, AND THE 1580 01:15:11,691 --> 01:15:12,892 OTHERS ARE LESS FREQUENT THAN 1581 01:15:12,892 --> 01:15:15,261 THAT BUT THEY'RE ALL HIGHLY 1582 01:15:15,261 --> 01:15:16,262 MUTAGENIC AND VERY, VERY 1583 01:15:16,262 --> 01:15:17,897 INTERESTING TO PEOPLE INTERESTED 1584 01:15:17,897 --> 01:15:20,066 IN MUTAGENESIS LIKE I AM. 1585 01:15:20,066 --> 01:15:23,002 AND SO WE GAVE ALL THOSE 1586 01:15:23,002 --> 01:15:24,337 LITTLE -- THOSE NUMBERS █OVER 1587 01:15:24,337 --> 01:15:26,172 THERE, LITTLE DOTS, DESCRIBING 1588 01:15:26,172 --> 01:15:28,308 THE ABUNDANCE OF THAT KIND OF 1589 01:15:28,308 --> 01:15:31,511 DAMAGE IN A CELL, AND WHEN MY 1590 01:15:31,511 --> 01:15:33,046 POSTDOC AND HER STUDENT WORKED 1591 01:15:33,046 --> 01:15:37,483 ON HOW FREQUENTLY THE REPLICASES 1592 01:15:37,483 --> 01:15:40,219 INCORPORATED RIBONUCLEOTIDE 1593 01:15:40,219 --> 01:15:41,321 TRIPHOSPHATES INTO THE NUCLEAR 1594 01:15:41,321 --> 01:15:43,122 GENOME DURING NORMAL REPLICATION 1595 01:15:43,122 --> 01:15:47,393 IN YEAST, NO DAMAGE, THAT'S WHAT 1596 01:15:47,393 --> 01:15:49,896 WE SAW. 1597 01:15:49,896 --> 01:15:52,198 OF ALL THE KINDS OF 1598 01:15:52,198 --> 01:15:53,099 NON-CANONICAL NUCLEOTIDES PUT 1599 01:15:53,099 --> 01:15:56,202 INTO THE GENOME IN EUKARYOTES, 1600 01:15:56,202 --> 01:16:00,306 THE MOST ABUNDANT ARE 1601 01:16:00,306 --> 01:16:01,407 RIBONUCLEOTIDE TRIPHOSPHATES. 1602 01:16:01,407 --> 01:16:02,709 SO THAT'S LED TO A LOT OF WORK 1603 01:16:02,709 --> 01:16:04,577 THAT WE AND OTHERS HAVE DONE AND 1604 01:16:04,577 --> 01:16:08,114 NOW THE NUMBER OF PEOPLE WORKING 1605 01:16:08,114 --> 01:16:10,116 ON RIBONUCLEOTIDE INCORPORATION 1606 01:16:10,116 --> 01:16:11,517 DURING REPLICATION IS MUCH, MUCH 1607 01:16:11,517 --> 01:16:14,220 LARGER THAN IT WAS IN 2010, WHEN 1608 01:16:14,220 --> 01:16:20,326 WE PUBLISHED THIS RESULT IN THE 1609 01:16:20,326 --> 01:16:23,596 PNAS, AND WE'VE DONE TWO THINGS 1610 01:16:23,596 --> 01:16:24,030 WITH IT. 1611 01:16:24,030 --> 01:16:26,099 WE'VE USED RIBONUCLEOTIDE 1612 01:16:26,099 --> 01:16:27,200 INCORPORATION IN A YEAST CELL 1613 01:16:27,200 --> 01:16:29,769 THAT COULD NOT REPAIR THOSE 1614 01:16:29,769 --> 01:16:31,204 RIBONUCLEOTIDE, AND WE FOUND 1615 01:16:31,204 --> 01:16:33,039 THAT USING THIS NEW PROCEDURE 1616 01:16:33,039 --> 01:16:35,675 THAT WE DEVELOPED A LONG TIME 1617 01:16:35,675 --> 01:16:42,849 AGO, HYDEN-SEQ, WHEN WE MAP 1618 01:16:42,849 --> 01:16:44,450 WHERE RIBONUCLEOTIDES ARE PUT 1619 01:16:44,450 --> 01:16:45,418 IN, THEY'RE ENTIRELY CON SITION 1620 01:16:45,418 --> 01:16:47,053 TENT WITH OUR MODEL FROM THE 1621 01:16:47,053 --> 01:16:48,888 MUTAGENESIS STUDIES WHICH SHOW 1622 01:16:48,888 --> 01:16:49,656 THAT RIBONUCLEOTIDES ARE 1623 01:16:49,656 --> 01:16:52,825 FREQUENTLY PUT IN BY POL ALPHA, 1624 01:16:52,825 --> 01:16:54,594 LESS FREQUENTLY BY POL DELTA, 1625 01:16:54,594 --> 01:16:56,696 AND LESS FREQUENTLY STILL BY POL 1626 01:16:56,696 --> 01:16:59,666 EPSILON, BUT THEY'RE STILL 1627 01:16:59,666 --> 01:17:01,968 ABUNDANTLY MISINCORPORATED INTO 1628 01:17:01,968 --> 01:17:04,604 THE NUCLEAR GENOME, AND WHERE 1629 01:17:04,604 --> 01:17:08,808 THEY ARE PUT IN IS ENTIRELY 1630 01:17:08,808 --> 01:17:09,842 CONSISTENT WITH OUR MODEL THAT'S 1631 01:17:09,842 --> 01:17:12,845 NOW IN TEXTBOOKS OF WHICH ROLES 1632 01:17:12,845 --> 01:17:15,782 THOSE THREE REPLICASES PERFORM 1633 01:17:15,782 --> 01:17:17,116 DURING DNA REPLICATION. 1634 01:17:17,116 --> 01:17:22,822 WE'VE ALSO ASKED WHAT ARE THE 1635 01:17:22,822 --> 01:17:23,656 CONSEQUENCES OF RIBONUCLEOTIDES 1636 01:17:23,656 --> 01:17:23,956 IN DNA. 1637 01:17:23,956 --> 01:17:27,593 YOU CAN GO TO THAT ANNUAL REVIEW 1638 01:17:27,593 --> 01:17:28,161 OR BIOCHEMISTRY REVIEW WE 1639 01:17:28,161 --> 01:17:30,997 PUBLISHED A COUPLE YEARS AGO AND 1640 01:17:30,997 --> 01:17:34,500 IT DESCRIBES A LOT OF BIOLOGY, 1641 01:17:34,500 --> 01:17:38,971 GENETICS, AND NOW GENOMICS THAT 1642 01:17:38,971 --> 01:17:40,940 DESCRIBES HOW MANY RIBOS ARE PUT 1643 01:17:40,940 --> 01:17:42,475 IN DURING REPLICATION AND HOW 1644 01:17:42,475 --> 01:17:44,277 THEY ARE REPAIRED, AND THEY ARE 1645 01:17:44,277 --> 01:17:46,345 REPAIRED BY A PROCESS WHICH WE 1646 01:17:46,345 --> 01:17:49,916 NOW CALL RIBONUCLEOTIDE EXCISION 1647 01:17:49,916 --> 01:17:51,584 REPAIR, WHICH IS IN ADDITION TO 1648 01:17:51,584 --> 01:17:53,319 MISMATCH REPAIR, A STRONG 1649 01:17:53,319 --> 01:17:54,754 PROTECTOR OF PUTTING THE WRONG 1650 01:17:54,754 --> 01:17:58,124 THING INTO THE GENOME. 1651 01:17:58,124 --> 01:17:59,759 AND WHEN WE DO THAT, AND I 1652 01:17:59,759 --> 01:18:08,267 FORGOT WHAT I DID HERE, IF YOU 1653 01:18:08,267 --> 01:18:12,171 KNOCK OUT RNA H2 IN HUMANS OR 1654 01:18:12,171 --> 01:18:15,041 YEAST, YOU HAVE LOTS OF 1655 01:18:15,041 --> 01:18:16,409 RIBONUCLEOTIDES THAT WERE 1656 01:18:16,409 --> 01:18:17,310 INADVERTENTLY INCORPORATED 1657 01:18:17,310 --> 01:18:20,580 DURING REPLICATION, AND WHEN 1658 01:18:20,580 --> 01:18:26,152 THEY'RE NOT REMOVED, BECAUSE OF 1659 01:18:26,152 --> 01:18:28,421 THE RNA STAGE II DEFECT, THEY 1660 01:18:28,421 --> 01:18:29,522 CAUSE MUTAGENESIS, WHICH IS ONE 1661 01:18:29,522 --> 01:18:30,656 OF THE MAIN THINGS THAT MY LAB 1662 01:18:30,656 --> 01:18:31,357 IS INTERESTED IN. 1663 01:18:31,357 --> 01:18:32,892 SO IF YOU'RE INTERESTED, YOU CAN 1664 01:18:32,892 --> 01:18:35,595 READ THAT ARTICLE. 1665 01:18:35,595 --> 01:18:37,897 AND WE'RE VERY PROUD OF THIS 1666 01:18:37,897 --> 01:18:40,533 WORK BECAUSE FOR 50 YEARS, IT 1667 01:18:40,533 --> 01:18:44,203 WAS BIOCHEMISTS I TELLING US 1668 01:18:44,203 --> 01:18:45,104 RIBONUCLEOTIDES WERE 1669 01:18:45,104 --> 01:18:46,072 INCORPORATED DURING REPLICATION 1670 01:18:46,072 --> 01:18:50,076 BUT ONLY RARELY, TO IN 2010, 1671 01:18:50,076 --> 01:18:53,646 DISCOVERING THAT THEY'RE NOT 1672 01:18:53,646 --> 01:18:57,483 THAT RARE AND THAT IMPLIES THERE 1673 01:18:57,483 --> 01:18:58,918 MAY BE SOME IMPORTANT SELECTIVE 1674 01:18:58,918 --> 01:19:01,320 ADVANTAGES TO INCORPORATING 1675 01:19:01,320 --> 01:19:02,688 RIBONUCLEOTIDES INTO DNA, ONE OF 1676 01:19:02,688 --> 01:19:05,525 WHICH WE PROPOSE AS A STRAND 1677 01:19:05,525 --> 01:19:07,627 SIGNAL DISCRIMINATION FOR DNA 1678 01:19:07,627 --> 01:19:08,628 MISMATCH REPAIR. 1679 01:19:08,628 --> 01:19:10,997 SO THIS IS EUKARYOTIC MISMATCH 1680 01:19:10,997 --> 01:19:12,064 REPAIR, AND YOU CAN SEE GOING 1681 01:19:12,064 --> 01:19:13,332 FROM THE TOP, WHERE A MISMATCH 1682 01:19:13,332 --> 01:19:20,373 IS IN THAT LITTLE HEX AGOAL BOX, 1683 01:19:20,373 --> 01:19:22,341 THAT'S LEFT BEHIND BY THE 1684 01:19:22,341 --> 01:19:23,476 REPLICATION FORK AND IT NEEDS TO 1685 01:19:23,476 --> 01:19:25,144 BE REPAIRED OR YOU GET MUTATIONS 1686 01:19:25,144 --> 01:19:26,345 THAT AMONG OTHER THINGS CAUSE 1687 01:19:26,345 --> 01:19:28,414 CANCER, AND THE STEPS THAT ARE 1688 01:19:28,414 --> 01:19:30,049 INVOLVED ARE A NUMBER OF 1689 01:19:30,049 --> 01:19:34,754 PROTEINS ULTIMATELY LEADING TO A 1690 01:19:34,754 --> 01:19:35,755 CORRECT MOLECULE AT THE BOTTOM 1691 01:19:35,755 --> 01:19:37,156 IN WHICH THAT MISMATCH REPAIR 1692 01:19:37,156 --> 01:19:42,762 HAS BEEN REMOVED. 1693 01:19:42,762 --> 01:19:45,364 SO WE'VE BEEN STUDYING THAT AS 1694 01:19:45,364 --> 01:19:48,668 WELL FOR QUITE SOME TIME, EVEN 1695 01:19:48,668 --> 01:19:50,369 BEFORE THE NOBEL PRIZE FOR 1696 01:19:50,369 --> 01:19:52,104 STUDYING MISMATCH REPAIR BACK IN 1697 01:19:52,104 --> 01:19:54,106 2015. 1698 01:19:54,106 --> 01:19:57,810 AND WE'VE BEEN STUDYING IT BASED 1699 01:19:57,810 --> 01:20:02,048 ON AN ARTICLE PUBLISHED IN THE 1700 01:20:02,048 --> 01:20:06,185 PNAS BY MAT MESSELSON AT MIT WHO 1701 01:20:06,185 --> 01:20:07,486 WROTE IN 1976 THAT IF YOU WANT 1702 01:20:07,486 --> 01:20:09,021 TO UNDERSTAND THE RELAWTION SHIP 1703 01:20:09,021 --> 01:20:12,425 BETWEEN REPLICATION AND DNA 1704 01:20:12,425 --> 01:20:13,793 MISMATCH REPAIR IN VIVO, IT MAY 1705 01:20:13,793 --> 01:20:17,730 BE POSSIBLE THAT MISMATCH REPAIR 1706 01:20:17,730 --> 01:20:21,300 ACTS IN A DIRECTED M MANNER, 1707 01:20:21,300 --> 01:20:22,935 POSSIBLY BECAUSE OF THE SPECIAL 1708 01:20:22,935 --> 01:20:23,936 RELATIONSHIP TO THE REPLICATION 1709 01:20:23,936 --> 01:20:24,370 COMPLEX. 1710 01:20:24,370 --> 01:20:25,538 MY LAB, WHICH IS VERY INTERESTED 1711 01:20:25,538 --> 01:20:27,306 IN MECHANISMS THAT DETERMINE 1712 01:20:27,306 --> 01:20:29,575 REPLICATION FIDELITY, HAS BEEN 1713 01:20:29,575 --> 01:20:31,878 INVESTIGATING THOSE PROCESSES. 1714 01:20:31,878 --> 01:20:34,280 DOES MISMATCH REPAIR EFFICIENCY 1715 01:20:34,280 --> 01:20:36,349 DEPEND ON ANYTHING DESCRIBED IN 1716 01:20:36,349 --> 01:20:37,016 BLACK BELOW THAT. 1717 01:20:37,016 --> 01:20:39,919 AND WE'VE DONE KUWAI QUITE A BIF 1718 01:20:39,919 --> 01:20:41,454 WORK ON THAT OVER THE YEARS, AND 1719 01:20:41,454 --> 01:20:43,322 I JUST WANT TO END BY SHOWING 1720 01:20:43,322 --> 01:20:46,893 YOU JUST A COUPLE OF THINGS. 1721 01:20:46,893 --> 01:20:48,961 THIS IS A SUMMARY SLIDE OF WHAT 1722 01:20:48,961 --> 01:20:51,163 I'VE SAID. 1723 01:20:51,163 --> 01:20:54,634 WHERE WE BELIEVE THE REPLICATION 1724 01:20:54,634 --> 01:20:56,369 FIDELITY OF THE NUCLEAR 1725 01:20:56,369 --> 01:21:00,640 REPLICASES ARE THOSE NUMBERS 1726 01:21:00,640 --> 01:21:02,608 COLORED IN COLORS UP THERE FOR 1727 01:21:02,608 --> 01:21:03,342 BASE SUBSTITUTIONS. 1728 01:21:03,342 --> 01:21:05,211 WE ALSO HAVE DONE A LOT OF WORK 1729 01:21:05,211 --> 01:21:08,180 ON INSERTION/DELETION MUTATIONS. 1730 01:21:08,180 --> 01:21:09,982 WE ALSO BELIEVE THAT 1731 01:21:09,982 --> 01:21:11,617 RIBONUCLEOTIDES ARE ABUNDANTLY 1732 01:21:11,617 --> 01:21:13,252 INCORPORATED DURING REPLICATION. 1733 01:21:13,252 --> 01:21:14,887 BUT BOTH OF THOSE KINDS OF 1734 01:21:14,887 --> 01:21:17,056 LESIONS CAN BE REPAIRED BY 1735 01:21:17,056 --> 01:21:19,926 EITHER MISMATCH REPAIR OR 1736 01:21:19,926 --> 01:21:21,093 RIBONUCLEOTIDE EXCISION REPAIR, 1737 01:21:21,093 --> 01:21:23,262 AND WE PUBLISHED A PAPER IN 1738 01:21:23,262 --> 01:21:24,563 MOLECULAR CELL A WHILE AGO 1739 01:21:24,563 --> 01:21:33,506 SHOWING THAT NUCLEOTIDE NUCLEON 1740 01:21:33,506 --> 01:21:35,441 REPAIR -- TO CREATE A NIC DNA 1741 01:21:35,441 --> 01:21:36,776 AND THAT NIC CAN POSSIBLY BE 1742 01:21:36,776 --> 01:21:38,945 USED TO REPAIR REPLICATION 1743 01:21:38,945 --> 01:21:41,547 ERRORS BY MISMATCH REPAIR. 1744 01:21:41,547 --> 01:21:43,416 THAT'S A SUMMARY OF WHAT WE'VE 1745 01:21:43,416 --> 01:21:43,616 DONE. 1746 01:21:43,616 --> 01:21:44,951 NOW I WANT TO SHOW YOU JUST A 1747 01:21:44,951 --> 01:21:47,553 LITTLE BIT MORE. 1748 01:21:47,553 --> 01:21:50,690 IN THE COURSE OF THIS WORK, BACK 1749 01:21:50,690 --> 01:21:53,859 IN 1985 WHEN I HAD BEEN RUNNING 1750 01:21:53,859 --> 01:21:56,262 MY OWN SHOW, WHICH IS ME AND ONE 1751 01:21:56,262 --> 01:22:00,499 TECHNICIAN TO BEGIN WITH, AT THE 1752 01:22:00,499 --> 01:22:03,502 NIEHS, I DISCOVERED A METHOD 1753 01:22:03,502 --> 01:22:06,272 THAT COULD BE USED TO ENGINEER 1754 01:22:06,272 --> 01:22:10,009 ANY DNA SEQUENCE TO CHANGE ITS 1755 01:22:10,009 --> 01:22:11,544 NUCLEOTIDE COMPOSITION TO MAKE 1756 01:22:11,544 --> 01:22:12,745 MUTATIONS IN GENES. 1757 01:22:12,745 --> 01:22:19,685 AND I PUBLISHED THAT IN THE PNAS 1758 01:22:19,685 --> 01:22:22,855 IN 1985, RIGHT AFTER MY LAB GOT 1759 01:22:22,855 --> 01:22:23,089 STARTED. 1760 01:22:23,089 --> 01:22:25,725 AND I PUT THAT UP THERE NOT TO 1761 01:22:25,725 --> 01:22:27,159 BRAG, ALTHOUGH THAT'S ONE OF THE 1762 01:22:27,159 --> 01:22:28,961 REASONS THAT I THINK I GOT 1763 01:22:28,961 --> 01:22:31,597 ELECTED TO THE NATIONAL ACADEMY, 1764 01:22:31,597 --> 01:22:34,433 BUT IT'S BECAUSE WITHIN ABOUT 1765 01:22:34,433 --> 01:22:36,402 FIVE YEARS, I DID A LITERATURE 1766 01:22:36,402 --> 01:22:38,904 SEARCH AND IT HAD ALREADY 1767 01:22:38,904 --> 01:22:41,440 RECEIVED MORE THAN 10,000 1768 01:22:41,440 --> 01:22:42,241 CITATIONS. 1769 01:22:42,241 --> 01:22:46,312 SO IT'S ONE OF THE TOP FEW 1770 01:22:46,312 --> 01:22:50,016 PUBLICATIONS THE NIH HAS EVER 1771 01:22:50,016 --> 01:22:51,317 PUBLISHED IN TERMS OF THE NUMBER 1772 01:22:51,317 --> 01:22:53,753 OF CITATIONS, AND THAT WAS 1773 01:22:53,753 --> 01:22:55,287 BECAUSE IT WAS -- THAT PROCEDURE 1774 01:22:55,287 --> 01:22:58,524 WAS SO APPLICABLE TO MANY, MANY 1775 01:22:58,524 --> 01:23:02,128 QUESTIONS IN BIOLOGY WHERE YOU 1776 01:23:02,128 --> 01:23:03,562 WANT TO MAKE A MUTATION IN THE 1777 01:23:03,562 --> 01:23:05,631 GENE, BUT IT WAS DONE IN 1985, 1778 01:23:05,631 --> 01:23:06,932 WHEN IT WAS VERY DIFFICULT TO DO 1779 01:23:06,932 --> 01:23:09,301 UNLESS YOU HAD SOME PHENOTYPIC 1780 01:23:09,301 --> 01:23:09,835 SLEK SHUP. 1781 01:23:09,835 --> 01:23:12,405 SELECTION. 1782 01:23:12,405 --> 01:23:14,740 THIS ALLOWED ME TO DO THAT AS 1783 01:23:14,740 --> 01:23:18,577 ONE EVENT OUT OF EVERY TWO 1784 01:23:18,577 --> 01:23:21,514 GENOMES THAT I SEQUENCED. 1785 01:23:21,514 --> 01:23:22,415 WITHOUT ANY PHENOTYPIC 1786 01:23:22,415 --> 01:23:22,948 SELECTION. 1787 01:23:22,948 --> 01:23:24,683 YOU CAN READ THAT PAPER IF 1788 01:23:24,683 --> 01:23:25,351 YOU'RE INTERESTED. 1789 01:23:25,351 --> 01:23:27,987 BUT IT MADE ME THINK VERY 1790 01:23:27,987 --> 01:23:29,188 STRONGLY LAST WEEK WHEN A 1791 01:23:29,188 --> 01:23:31,557 COLLEAGUE OF MINE SENT ME A 1792 01:23:31,557 --> 01:23:32,425 PUBLICATION AND SAID YOU'VE GOT 1793 01:23:32,425 --> 01:23:36,262 TO READ THIS, AND IT WAS APPLIED 1794 01:23:36,262 --> 01:23:38,431 RESEARCH WON'T FLOURISH WITHOUT 1795 01:23:38,431 --> 01:23:40,299 BASIC SCIENCE, AND THIS WAS BY 1796 01:23:40,299 --> 01:23:44,336 THE THREE AUTHORS LISTED THERE, 1797 01:23:44,336 --> 01:23:47,273 WHO DESCRIBED THE PORTFOLIO OF 1798 01:23:47,273 --> 01:23:49,675 RESEARCH THAT NIH HAS FUNDED 1799 01:23:49,675 --> 01:23:53,379 SINCE ITS EXISTENCE, AND DECIDED 1800 01:23:53,379 --> 01:23:57,550 THAT ABOUT 51% OF THAT RESEARCH 1801 01:23:57,550 --> 01:23:58,751 IS CURRENTLY BASIC RESEARCH, 1802 01:23:58,751 --> 01:24:03,222 WHICH IS WHAT MY LAB STUDIES, 1803 01:24:03,222 --> 01:24:05,958 VERY BASIC QUESTIONS, AND ABOUT 1804 01:24:05,958 --> 01:24:10,196 49% IS CURRENTLY IN WHAT I CALL 1805 01:24:10,196 --> 01:24:11,397 ARTIFICIALLY TRANSLATIONAL 1806 01:24:11,397 --> 01:24:13,466 RESEARCH AS BEAUTIFULLY 1807 01:24:13,466 --> 01:24:14,333 ILLUSTRATED BY THE FIRST TALK WE 1808 01:24:14,333 --> 01:24:19,472 HEARD HERE. 1809 01:24:19,472 --> 01:24:20,906 SO WITH THAT THOUGHT IN MIND, I 1810 01:24:20,906 --> 01:24:22,408 WANTED TO END THIS BY DESCRIBING 1811 01:24:22,408 --> 01:24:23,609 TO YOU ONE THING THAT WE'RE 1812 01:24:23,609 --> 01:24:26,011 REALLY EXCITED ABOUT NOW IN MY 1813 01:24:26,011 --> 01:24:26,345 LAB. 1814 01:24:26,345 --> 01:24:28,647 I'M INTERESTED IN THE QUESTION 1815 01:24:28,647 --> 01:24:33,886 OF DOES SIMPLE CHAIN ELONGATION 1816 01:24:33,886 --> 01:24:36,922 COPY DNA WITH VERY, VERY HIGH 1817 01:24:36,922 --> 01:24:39,992 FIDELITY, WHEREAS STRAND 1818 01:24:39,992 --> 01:24:42,061 DISPLACEMENT, WHICH OCCURRED 1819 01:24:42,061 --> 01:24:45,998 DURING OKAZAKI FRAGMENT 1820 01:24:45,998 --> 01:24:48,067 MATURATION, THAT REQUIRES PRIMER 1821 01:24:48,067 --> 01:24:49,468 B BE REMOVED FROM THE DNA AND 1822 01:24:49,468 --> 01:24:50,803 THAT'S DONE, WELL, I'LL SHOW YOU 1823 01:24:50,803 --> 01:24:52,738 HOW THAT'S DONE IN THE NEXT 1824 01:24:52,738 --> 01:24:53,973 SLIDE, BUT GOING BACK TO ONE OF 1825 01:24:53,973 --> 01:24:55,474 THOSE SLIDES I SHOWED IN THE 1826 01:24:55,474 --> 01:24:57,009 MIDDLE, THE ELONGATION ON AN 1827 01:24:57,009 --> 01:24:59,311 OPEN TEMPLATE HAS TO OCCUR FOR 1828 01:24:59,311 --> 01:25:00,713 EVERY NUCLEOTIDE THAT'S 1829 01:25:00,713 --> 01:25:03,883 INCORPORATED OR MOST EVERY 1830 01:25:03,883 --> 01:25:05,951 NUCLEOTIDE, BUT OKAZAKI FRAGMENT 1831 01:25:05,951 --> 01:25:08,020 MATURATION USES THE LOWER THING, 1832 01:25:08,020 --> 01:25:09,321 STRAND DISPLACEMENT. 1833 01:25:09,321 --> 01:25:11,423 SO WE'RE VERY INTERESTED IN THAT 1834 01:25:11,423 --> 01:25:13,058 QUESTION, ENERGETICALLY, 1835 01:25:13,058 --> 01:25:14,994 BIOCHEMICALLY, STRUCTURALLY, BUT 1836 01:25:14,994 --> 01:25:17,062 WE'RE ALSO INTERESTED IN THAT 1837 01:25:17,062 --> 01:25:19,698 FROM THE STANDPOINT OF APPLYING 1838 01:25:19,698 --> 01:25:22,101 WHOLE GENOME SEQUENCING GENOMICS 1839 01:25:22,101 --> 01:25:25,804 APPROACH TO THE QUESTION. 1840 01:25:25,804 --> 01:25:29,642 SO WE DID THAT IN A YEAST STRAIN 1841 01:25:29,642 --> 01:25:33,012 THAT LACKS OKAZAKI FRAGMENT 1842 01:25:33,012 --> 01:25:35,948 MATURATION PROCESSING BECAUSE IT 1843 01:25:35,948 --> 01:25:38,184 LACKS FLAP ENDO NUCLEASE 1, 1844 01:25:38,184 --> 01:25:40,219 WHICH DISRUPTS THAT FLAP, SO 1845 01:25:40,219 --> 01:25:44,690 THAT IT CAN BE EXCISED BY FEN1, 1846 01:25:44,690 --> 01:25:48,827 AND WHEN WE DID THAT MONTHS AGO, 1847 01:25:48,827 --> 01:25:50,996 WE FOUND SOMETHING THAT TOTALLY 1848 01:25:50,996 --> 01:25:52,865 CONFUSES ME. 1849 01:25:52,865 --> 01:25:55,134 BUT IN A GOOD WAY. 1850 01:25:55,134 --> 01:25:57,870 IT'S ALWAYS GREAT TO BE 1851 01:25:57,870 --> 01:26:01,473 CONFUSED, BECAUSE EVENTUALLY IF 1852 01:26:01,473 --> 01:26:02,474 YOU WORK HARD ENOUGH ON IT, YOU 1853 01:26:02,474 --> 01:26:04,109 CAN FIND OUT SOMETHING 1854 01:26:04,109 --> 01:26:09,148 INTERESTING. 1855 01:26:09,148 --> 01:26:18,857 SO -- I APOLOGIZE FOR THAT. 1856 01:26:18,857 --> 01:26:20,025 I JUST WANT TO SHOW YOU TWO 1857 01:26:20,025 --> 01:26:20,292 SLIDES. 1858 01:26:20,292 --> 01:26:23,229 WHEN WE INVESTIGATED MISMATCH 1859 01:26:23,229 --> 01:26:24,430 REPAIR, IF I CAN EVER GET BACK 1860 01:26:24,430 --> 01:26:34,940 TO THAT, ONE OF THE NUCLEASES 1861 01:26:35,975 --> 01:26:37,409 THAT'S THOUGHT TO BE INVOLVED IN 1862 01:26:37,409 --> 01:26:39,511 REMOVING A MISMATCH, ONCE THE 1863 01:26:39,511 --> 01:26:41,614 NEWLY SYNTHESIZED STRAND HAS 1864 01:26:41,614 --> 01:26:45,451 BEEN NICKED TO INCLUDE AN ENTRY 1865 01:26:45,451 --> 01:26:52,157 POINT FOR A NUCLEASE, ONE OF THE 1866 01:26:52,157 --> 01:26:54,560 NUCLEASES IS FEN1 THAT WORKS 1867 01:26:54,560 --> 01:26:56,295 DURING OKAZAKI FRAGMENT 1868 01:26:56,295 --> 01:26:56,929 MATURATION, AND THAT'S WHAT I 1869 01:26:56,929 --> 01:27:01,433 WAS SHOWING HERE. 1870 01:27:01,433 --> 01:27:04,169 THIS, HOWEVER, IS A VERY 1871 01:27:04,169 --> 01:27:04,937 COMPLICATED SLIDE SO I'M NOT 1872 01:27:04,937 --> 01:27:06,338 GOING TO TRY TO EXPLAIN IT TOO 1873 01:27:06,338 --> 01:27:08,307 MUCH, BUT TO TRY TO SAY THAT 1874 01:27:08,307 --> 01:27:09,608 THIS IS THE CHANGE IN LENGTH IN 1875 01:27:09,608 --> 01:27:13,312 BASE PAIRS, EITHER INSERTIONS ON 1876 01:27:13,312 --> 01:27:15,614 THE RIGHT OR DELETIONS ON THE 1877 01:27:15,614 --> 01:27:18,784 LEFT, WHICH OCCURS IN A STRAIN 1878 01:27:18,784 --> 01:27:22,821 THAT'S LACKING MISMATCH REPAIR 1879 01:27:22,821 --> 01:27:23,889 COMPLETELY BECAUSE YOU'VE 1880 01:27:23,889 --> 01:27:25,324 DISRUPTED THE FIRST SEMINAL 1881 01:27:25,324 --> 01:27:30,929 PROTEIN IN THE PATHWAY, THE MUSE 1882 01:27:30,929 --> 01:27:32,264 S ALPHA HETERODIMER. 1883 01:27:32,264 --> 01:27:34,400 AND IN GREEN YOU SEE ALL THE 1884 01:27:34,400 --> 01:27:36,268 SINGLE BASE MUTATIONS CREATED IN 1885 01:27:36,268 --> 01:27:38,771 AN MSH2 STRAIN, ONE OF THE 1886 01:27:38,771 --> 01:27:40,072 REASONS IT'S SUSPECTED IN BEING 1887 01:27:40,072 --> 01:27:41,273 HEAVILY INVOLVED IN TUMOR 1888 01:27:41,273 --> 01:27:42,508 FORMATION. 1889 01:27:42,508 --> 01:27:45,110 AND AS YOU GO UP AND OUT, YOU 1890 01:27:45,110 --> 01:27:48,614 SEE THERE'S MUCH LESS LOWER RATE 1891 01:27:48,614 --> 01:27:53,185 OF REPLICATION ERRORS IN A 1892 01:27:53,185 --> 01:27:55,788 MISMATCH REPAIR DEFECTIVE YEAST 1893 01:27:55,788 --> 01:27:58,991 CELL AS ILLUSTRATED BY THE RED, 1894 01:27:58,991 --> 01:28:00,726 WHICH HAVE -- IF YOU COULD READ 1895 01:28:00,726 --> 01:28:02,428 IT, YOU CAN'T, LOWER MUTATION 1896 01:28:02,428 --> 01:28:02,895 RATES. 1897 01:28:02,895 --> 01:28:04,396 SO THE PREDOMINANT CONSEQUENCE 1898 01:28:04,396 --> 01:28:07,599 OF KNOCKING OUT ALL MISMATCH 1899 01:28:07,599 --> 01:28:09,802 REPAIR IS POINT MUTATIONS. 1900 01:28:09,802 --> 01:28:11,503 AND WE PUBLISHED THIS QUITE A 1901 01:28:11,503 --> 01:28:13,038 WHILE AGO, BUT I PUT IT LIKE 1902 01:28:13,038 --> 01:28:14,373 THIS BECAUSE THIS IS WHAT 1903 01:28:14,373 --> 01:28:19,912 HAPPENED WHEN WE KNOCKED OUT RAD 1904 01:28:19,912 --> 01:28:20,245 27. 1905 01:28:20,245 --> 01:28:21,914 SO THERE ARE SO MANY THINGS 1906 01:28:21,914 --> 01:28:25,050 DIFFERENT ABOUT THIS SLIDE THAN 1907 01:28:25,050 --> 01:28:27,553 KNOCKING OUT MISMATCH REPAIR, IT 1908 01:28:27,553 --> 01:28:31,623 REALLY CONFUSES ME, BUT YOU 1909 01:28:31,623 --> 01:28:32,157 STILL SEE GREEN. 1910 01:28:32,157 --> 01:28:36,428 A LOT OF FRAME SHIFT MUTATIONS 1911 01:28:36,428 --> 01:28:38,163 RESULTING FROM LOSS OF DNA 1912 01:28:38,163 --> 01:28:39,798 MISMATCH REPAIR, SO RAD 27 IS 1913 01:28:39,798 --> 01:28:41,567 INVOLVED IN THAT PROCESS, BUT 1914 01:28:41,567 --> 01:28:45,237 YOU ALSO SEE A LOT OF OTHER 1915 01:28:45,237 --> 01:28:48,440 ERRORS IN VARIOUS ERRORS AROUND 1916 01:28:48,440 --> 01:28:50,275 THIS CURVE, THEY INVOLVE BOTH 1917 01:28:50,275 --> 01:28:52,010 DELETIONS AND INSERTIONS, SO WE 1918 01:28:52,010 --> 01:28:53,645 BELIEVE THERE'S AT LEAST ONE 1919 01:28:53,645 --> 01:28:57,816 OTHER MECHANISM THAT IS CRITICAL 1920 01:28:57,816 --> 01:29:03,489 THAT RAD 27 FLAP ENDONUCLEASE BE 1921 01:29:03,489 --> 01:29:04,223 THERE FOR BECAUSE OF THE HIGH 1922 01:29:04,223 --> 01:29:07,393 RATE OF MU TEATION MUTATIONS INR 1923 01:29:07,393 --> 01:29:08,060 REGIONS OF THAT PLOT. 1924 01:29:08,060 --> 01:29:09,061 WE DON'T UNDERSTAND THAT. 1925 01:29:09,061 --> 01:29:11,063 I THINK, AS DOES MY COLLEAGUE 1926 01:29:11,063 --> 01:29:13,198 WHO IS RESPONSIBLE FOR THIS 1927 01:29:13,198 --> 01:29:16,702 WORK, THAT THERE MAY BE A TOTAL 1928 01:29:16,702 --> 01:29:20,639 OF 30 DIFFERENT MECHANISMS 1929 01:29:20,639 --> 01:29:22,508 RESPONSIBLE FOR THIS, OF WHICH 1930 01:29:22,508 --> 01:29:24,109 ALL OF THEM ARE AFFECTED BY LOSS 1931 01:29:24,109 --> 01:29:26,945 OF RAD 27, WHICH ANOTHER WAY TO 1932 01:29:26,945 --> 01:29:32,084 SAY THAT IS IT'S DOING MUCH MORE 1933 01:29:32,084 --> 01:29:33,352 THAN CREATING POINT MUTATIONS, 1934 01:29:33,352 --> 01:29:34,820 WHICH IS THE MAJOR EFFECT OF 1935 01:29:34,820 --> 01:29:35,921 LOSS OF MISMATCH REPAIR. 1936 01:29:35,921 --> 01:29:37,956 I DON'T UNDERSTAND THAT EXCEPT, 1937 01:29:37,956 --> 01:29:39,191 I MEAN, I COULD GO ON FOR HALF 1938 01:29:39,191 --> 01:29:40,592 AN HOUR ABOUT THIS, BUT I'M NOT 1939 01:29:40,592 --> 01:29:42,995 GOING TO. 1940 01:29:42,995 --> 01:29:44,196 I AM GOING TO POINT OUT THAT IN 1941 01:29:44,196 --> 01:29:49,134 THE LAST FEW YEARS, MANY 1942 01:29:49,134 --> 01:29:51,203 ARTICLES HAVE DESCRIBED 1943 01:29:51,203 --> 01:29:52,337 SPECIFICITY IN TUMORS ISOLATED 1944 01:29:52,337 --> 01:29:53,806 FROM A WIDE VARIETY OF CANCERS. 1945 01:29:53,806 --> 01:29:58,210 EVERY TIME I OPEN YOU C UP CELL, 1946 01:29:58,210 --> 01:29:59,411 NATURE OR SCIENCE, I READ 1947 01:29:59,411 --> 01:30:00,712 ANOTHER VERY EXCITING ARTICLE ON 1948 01:30:00,712 --> 01:30:01,380 THAT SUBJECT. 1949 01:30:01,380 --> 01:30:02,815 THESE STUDIES FOCUS ON THE 1950 01:30:02,815 --> 01:30:05,083 IDENTITY OF POINT MUTATIONS THAT 1951 01:30:05,083 --> 01:30:08,787 DRIVE TUMOR FORMATION TOWARDS 1952 01:30:08,787 --> 01:30:10,022 DEFINING TARGETS FOR CLINICAL 1953 01:30:10,022 --> 01:30:11,423 INTERVENTION. 1954 01:30:11,423 --> 01:30:13,392 THE TRANSLATIONAL SIDE OF WHAT 1955 01:30:13,392 --> 01:30:15,360 NIH DOES. 1956 01:30:15,360 --> 01:30:16,995 THE RESULTS I JUST SHOWED YOU ON 1957 01:30:16,995 --> 01:30:21,800 THAT LAST SLIDE IN THE YEAST RAD 1958 01:30:21,800 --> 01:30:23,535 27 MUTATIONAL PROFILE SUGGESTS 1959 01:30:23,535 --> 01:30:25,838 THAT SUCH DRIVERS MAY INCLUDE 1960 01:30:25,838 --> 01:30:29,141 GENES, MUTATIONS IN GENES THAT 1961 01:30:29,141 --> 01:30:32,611 DRIVE LARGER MUTATIONAL EVENTS. 1962 01:30:32,611 --> 01:30:36,982 AND THAT SUBJECT HAS NOT YET 1963 01:30:36,982 --> 01:30:38,317 BEEN INVESTIGATED BY THOSE 1964 01:30:38,317 --> 01:30:39,618 MULTIPLE EXCITING TRANSLATIONAL 1965 01:30:39,618 --> 01:30:40,385 RESEARCH STUDIES. 1966 01:30:40,385 --> 01:30:43,822 SO THERE IS A WIDE OPPORTUNITY 1967 01:30:43,822 --> 01:30:46,825 FOR ANY OF YOU STUDENTS IN THE 1968 01:30:46,825 --> 01:30:48,126 AUDIENCE TO BECOME INVOLVED IN 1969 01:30:48,126 --> 01:30:52,498 THIS WHICH IS STILL A GROWTH 1970 01:30:52,498 --> 01:30:52,931 AREA. 1971 01:30:52,931 --> 01:30:54,399 AND LAST, I WANT TO SINCERELY 1972 01:30:54,399 --> 01:30:55,968 THANK THE MANY OUTSTANDING 1973 01:30:55,968 --> 01:30:57,402 COLLEAGUES THAT I'VE HAD OVER 1974 01:30:57,402 --> 01:30:59,805 THE YEARS FOR CONTRIBUTING TO 1975 01:30:59,805 --> 01:31:01,573 THIS RESEARCH FOR NOW 42 YEARS. 1976 01:31:01,573 --> 01:31:02,007 THANK YOU. 1977 01:31:02,007 --> 01:31:10,549 [APPLAUSE] 1978 01:31:10,549 --> 01:31:11,517 >> AGAIN, QUESTIONS? 1979 01:31:11,517 --> 01:31:12,684 I'M GOING TO CHECK ONLINE. 1980 01:31:12,684 --> 01:31:14,019 I KNOW THERE'S A LOT OF PEOPLE 1981 01:31:14,019 --> 01:31:15,287 WATCHING AND PERHAPS FROM NORTH 1982 01:31:15,287 --> 01:31:16,522 CAROLINA. 1983 01:31:16,522 --> 01:31:17,389 >> HELLO. 1984 01:31:17,389 --> 01:31:20,893 THANK YOU FOR YOUR INTERESTING 1985 01:31:20,893 --> 01:31:21,560 TALK. 1986 01:31:21,560 --> 01:31:23,095 I'M JUST WONDERING, ANY 1987 01:31:23,095 --> 01:31:24,596 PERSPECTIVE REGARDING TO AGING 1988 01:31:24,596 --> 01:31:26,999 AND TO FERTILITY? 1989 01:31:26,999 --> 01:31:29,535 >> YEAH, THERE ARE LOTS OF 1990 01:31:29,535 --> 01:31:31,603 STUDIES IN THE LITERATURE ON THE 1991 01:31:31,603 --> 01:31:39,912 ROLE OF MUTATIONS, OR FAILURE TO 1992 01:31:39,912 --> 01:31:41,780 CREATE MUTATIONS IN THE AGING 1993 01:31:41,780 --> 01:31:42,481 PROCESS. 1994 01:31:42,481 --> 01:31:44,783 VERY ROBUST SET OF LITERATURE. 1995 01:31:44,783 --> 01:31:48,654 IN FACT, NIH HAS AN AGING 1996 01:31:48,654 --> 01:31:49,655 INSTITUTE, AND SEVERAL OF MY 1997 01:31:49,655 --> 01:31:50,956 COLLEAGUES IN THAT INSTITUTE 1998 01:31:50,956 --> 01:31:53,258 COULD TELL YOU PLENTY MORE ABOUT 1999 01:31:53,258 --> 01:31:54,459 THAT SUBJECT THAN THAT BRIEF 2000 01:31:54,459 --> 01:32:03,335 ANSWER THAT I JUST GAVE. 2001 01:32:03,335 --> 01:32:08,674 >> IS THERE AN ANALOGOUS PROCESS 2002 01:32:08,674 --> 01:32:10,742 FOR WHEN A MISTAKE IS MADE IN 2003 01:32:10,742 --> 01:32:19,818 IRANIRNAELONGATION LIKE IN THE N 2004 01:32:19,818 --> 01:32:20,352 OF RNA? 2005 01:32:20,352 --> 01:32:23,855 >> YES, THERE ARE A LOT OF 2006 01:32:23,855 --> 01:32:26,458 ANALOGIES TO ERROR CORRECTION 2007 01:32:26,458 --> 01:32:29,962 DURING RNA SYNTHESIS AND ERROR 2008 01:32:29,962 --> 01:32:31,263 CORRECTION DURING DNA SYNTHESIS. 2009 01:32:31,263 --> 01:32:35,968 THERE'S A WORLD OF RESEARCH DONE 2010 01:32:35,968 --> 01:32:36,635 ON THAT SUBJECT. 2011 01:32:36,635 --> 01:32:41,740 OF COURSE ONE ERROR IN RNA 2012 01:32:41,740 --> 01:32:43,108 SYNTHESIS IS NOT AS SEVERE AS 2013 01:32:43,108 --> 01:32:45,577 ONE ERROR IN DNA SYNTHESIS 2014 01:32:45,577 --> 01:32:47,746 BECAUSE YOU MAKE A LOT MORE RNA 2015 01:32:47,746 --> 01:32:51,783 THAN YOU MAKE DNA, BUT 2016 01:32:51,783 --> 01:32:53,018 NONETHELESS, THERE ARE ANALOGIES 2017 01:32:53,018 --> 01:32:55,954 THAT HAVE BEEN DRAWN BETWEEN THE 2018 01:32:55,954 --> 01:32:56,321 TWO PROCESSES. 2019 01:32:56,321 --> 01:32:56,755 >> GREAT. 2020 01:32:56,755 --> 01:32:59,691 THANK YOU. 2021 01:32:59,691 --> 01:33:00,125 ALL RIGHT. 2022 01:33:00,125 --> 01:33:01,460 THANK YOU VERY, VERY MUCH. 2023 01:33:01,460 --> 01:33:04,162 [APPLAUSE] 2024 01:33:04,162 --> 01:33:05,597 I'VE BEEN ASKED BEFORE OUR FINAL 2025 01:33:05,597 --> 01:33:07,666 TALK JUST TO LET YOU KNOW THAT 2026 01:33:07,666 --> 01:33:10,936 THERE WILL BE A RECEPTION AFTER 2027 01:33:10,936 --> 01:33:14,439 THIS SESSION RIGHT IN THE LOBBY 2028 01:33:14,439 --> 01:33:15,073 DIRECTLY OUTSIDE. 2029 01:33:15,073 --> 01:33:18,276 AND OUR FINAL SPEAKER IS 2030 01:33:18,276 --> 01:33:18,844 DR. GIORGIO TRINCHIERI. 2031 01:33:18,844 --> 01:33:29,021 [APPLAUSE] 2032 01:33:30,055 --> 01:33:32,124 >> THANK YOU, THANK YOU VERY 2033 01:33:32,124 --> 01:33:32,324 MUCH. 2034 01:33:32,324 --> 01:33:35,093 I'M VERY HAPPY TO BE HERE, 2035 01:33:35,093 --> 01:33:36,628 REALLY EXCITED TO HEAR THE 2036 01:33:36,628 --> 01:33:37,496 PREVIOUS TWO TALKS. 2037 01:33:37,496 --> 01:33:38,697 I THINK MY TALK WILL HAVE 2038 01:33:38,697 --> 01:33:40,098 SOMETHING A LITTLE BIT DIFFERENT 2039 01:33:40,098 --> 01:33:42,200 THAT MY CAREER HAS BEEN NOT VERY 2040 01:33:42,200 --> 01:33:44,269 STABLE, I'VE BEEN MOVING AROUND, 2041 01:33:44,269 --> 01:33:45,737 I THINK A LOT MORE THAN THE TWO 2042 01:33:45,737 --> 01:33:48,740 PREVIOUS SPEAKERS. 2043 01:33:48,740 --> 01:33:55,847 AND I REALLY START IN ITALY, 2044 01:33:55,847 --> 01:34:00,185 THEN MOVED TO THE IMMUNOLOGY, 2045 01:34:00,185 --> 01:34:02,754 YOU CAME TO THE STATES IN 2046 01:34:02,754 --> 01:34:03,288 PHILADELPHIA. 2047 01:34:03,288 --> 01:34:06,258 I WENT BACK TO SWITZERLAND FOR A 2048 01:34:06,258 --> 01:34:11,063 COUPLE OF YEARS, AND THEN I 2049 01:34:11,063 --> 01:34:17,035 MOVED TO -- BACK TO WISTAR FOR 2050 01:34:17,035 --> 01:34:22,307 20 YEARS, I WENT TO A COMPANY 2051 01:34:22,307 --> 01:34:24,042 SCHERING-PLOUGH IN FRANCE, AND 2052 01:34:24,042 --> 01:34:25,711 IN 2004, COMING BACK FROM 2053 01:34:25,711 --> 01:34:27,446 INDUSTRY, I WAS VERY HAPPY TO 2054 01:34:27,446 --> 01:34:29,614 COME TO NIH. 2055 01:34:29,614 --> 01:34:31,283 FIRST I DID A SABBATICAL, 2056 01:34:31,283 --> 01:34:35,320 MENTIONED A LITTLE BIT OF THAT, 2057 01:34:35,320 --> 01:34:37,823 IN NIAID AND FOR ALMOST THE LAST 2058 01:34:37,823 --> 01:34:39,291 20 YEARS IN THE NATIONAL CANCER 2059 01:34:39,291 --> 01:34:41,126 INSTITUTE. 2060 01:34:41,126 --> 01:34:42,961 AND MY -- SO I'M GOING TO TRY TO 2061 01:34:42,961 --> 01:34:45,597 PUT TOGETHER MY HISTORY WITH 2062 01:34:45,597 --> 01:34:46,465 SOME OF THE SCIENCE THAT WAS 2063 01:34:46,465 --> 01:34:47,466 GOING AT THE SAME TIME, AND ONE 2064 01:34:47,466 --> 01:34:50,302 OF MY FIRST PROJECTS IN TORINO 2065 01:34:50,302 --> 01:34:53,905 BUT ALSO THE -- IMMUNOLOGY WAS 2066 01:34:53,905 --> 01:34:57,642 TO IDENTIFY WHAT THE CYTOTOXIC T 2067 01:34:57,642 --> 01:34:58,610 LYMPHOCYTE WOULD RECOGNIZE 2068 01:34:58,610 --> 01:35:02,013 WITHIN THE MAJOR 2069 01:35:02,013 --> 01:35:03,448 HISTOCOMPATIBILITY COMPLEX HLA 2070 01:35:03,448 --> 01:35:04,182 IN HUMAN. 2071 01:35:04,182 --> 01:35:06,218 SO OBVIOUSLY WE KNOW A LOT NOW 2072 01:35:06,218 --> 01:35:08,453 ABOUT HLA, CLASS 1, CLASS 2, 2073 01:35:08,453 --> 01:35:10,655 MANY DIFFERENT GENE, CLASS 3, 2074 01:35:10,655 --> 01:35:12,424 AND WE KNOW THE INVOLVEMENT IN 2075 01:35:12,424 --> 01:35:14,459 DISEASE AND ALL OF THAT. 2076 01:35:14,459 --> 01:35:17,195 BUT THAT WAS VERY DIFFERENT IN 2077 01:35:17,195 --> 01:35:22,267 THOSE YEARS, TALK ABOUT '68, 2078 01:35:22,267 --> 01:35:22,434 '72. 2079 01:35:22,434 --> 01:35:28,039 WE KNEW THAT THERE WERE ANTIGEN 2080 01:35:28,039 --> 01:35:31,676 THAT WERE RECOGNIZED BY SERA 2081 01:35:31,676 --> 01:35:33,845 FROM -- WOMAN AND THEY WERE 2082 01:35:33,845 --> 01:35:35,046 CALLED SEROLOGICAL DEFINE AND 2083 01:35:35,046 --> 01:35:36,248 JUST AT THAT TIME IT WAS 2084 01:35:36,248 --> 01:35:37,282 DISCOVERED THERE WERE ACTUALLY 2085 01:35:37,282 --> 01:35:38,984 TWO DIFFERENT LOCI, A AND B. 2086 01:35:38,984 --> 01:35:42,487 THESE ARE THE ANTIGEN THAT WERE 2087 01:35:42,487 --> 01:35:44,022 DESCRIBED ORIGINALLY BY SOME OF 2088 01:35:44,022 --> 01:35:48,827 THE PIONEER IN THE FIELD. 2089 01:35:48,827 --> 01:35:52,798 BUT ALSO FRITZ BARK AT THAT TIME 2090 01:35:52,798 --> 01:35:56,601 FOUND WHEN YOU MIX -- 2091 01:35:56,601 --> 01:35:58,470 TOGETHER -- CELL CULTURE AND 2092 01:35:58,470 --> 01:36:01,973 DEFINE A SEPARATE LOCUS IN THE 2093 01:36:01,973 --> 01:36:04,709 HLA CHROMOSOME THAT HE DEFINED 2094 01:36:04,709 --> 01:36:07,546 AS LYMPHOCYTE DEFINE NOW W WE KW 2095 01:36:07,546 --> 01:36:10,182 IS CLASS 2 MHC. 2096 01:36:10,182 --> 01:36:13,785 SO WHAT WE DID IS WE MIXED UP -- 2097 01:36:13,785 --> 01:36:16,721 CULTURE WITH STIMULATE -- 2098 01:36:16,721 --> 01:36:19,891 UNIDIRECTIONAL DONOR LYMPHOCYTE 2099 01:36:19,891 --> 01:36:20,892 STIMULATED BY THE LYMPHOCYTE 2100 01:36:20,892 --> 01:36:22,260 FROM DONOR B. 2101 01:36:22,260 --> 01:36:24,196 WE KNEW THAT WAS REGULATED BY 2102 01:36:24,196 --> 01:36:28,366 THIS LI LYMPHOCYTE DEFINED LOCU. 2103 01:36:28,366 --> 01:36:30,769 BUT WHEN WE LOOKED AT THE 2104 01:36:30,769 --> 01:36:32,204 CYTOTOXIC T-CELL IN THIS CULTURE 2105 01:36:32,204 --> 01:36:34,139 THEY WERE KILLING NOT FROM THE 2106 01:36:34,139 --> 01:36:35,574 LOCUS FROM WHICH THEY WERE 2107 01:36:35,574 --> 01:36:38,310 ACTIVATED BUT THEY RECOGNIZE THE 2108 01:36:38,310 --> 01:36:39,945 SEROLOGIC ANTIGEN -- THEY WERE 2109 01:36:39,945 --> 01:36:43,448 RECOGNIZED BY THE ANTIBODY ON 2110 01:36:43,448 --> 01:36:45,851 CLASS I HLA ORGANIZED BY THE 2111 01:36:45,851 --> 01:36:48,820 CYTOTOXIC T LYMPHOCYTE. 2112 01:36:48,820 --> 01:36:51,423 SO WE IDENTIFIED THAT THE T 2113 01:36:51,423 --> 01:36:53,491 LYMPHOCYTE RECOGNIZE CLASS 1 -- 2114 01:36:53,491 --> 01:36:56,261 CELL THAT OBVIOUSLY WAS 2115 01:36:56,261 --> 01:36:59,097 IMPORTANT FOR -- TRANSPLANT 2116 01:36:59,097 --> 01:37:00,832 AFTER GRAFT REJECTION. 2117 01:37:00,832 --> 01:37:06,404 A COUPLE OF YEARS LATER, 2118 01:37:06,404 --> 01:37:08,707 ZINGERNAGEL AND DOHERTY GOT 2119 01:37:08,707 --> 01:37:10,475 MAJOR FUNDING FOR WHICH THEY WON 2120 01:37:10,475 --> 01:37:13,578 A NOBEL PRIZE, THE RECOGNITION 2121 01:37:13,578 --> 01:37:15,413 OF VITAL ANTIGEN IS ACTUALLY 2122 01:37:15,413 --> 01:37:20,118 RESTRICTED BY SELF CLASS I MHC. 2123 01:37:20,118 --> 01:37:23,188 SO -- ALSO RECOGNIZE CLASS I 2124 01:37:23,188 --> 01:37:24,189 MHC. 2125 01:37:24,189 --> 01:37:25,390 WHAT WAS NOT KNOWN AT THE TIME, 2126 01:37:25,390 --> 01:37:26,791 THERE WAS A LOT OF DISCUSSION, 2127 01:37:26,791 --> 01:37:28,860 IS WHETHER A TUMOR ANTIGEN ALSO 2128 01:37:28,860 --> 01:37:30,729 NEEDED CLASS I FOR RECOGNITION. 2129 01:37:30,729 --> 01:37:35,533 SO WHEN I MOVED TO WISTAR, I 2130 01:37:35,533 --> 01:37:38,036 WORKED WITH BARB RECREATION AND 2131 01:37:38,036 --> 01:37:44,276 PARKS -- THAT WERE -- T ANTIGEN, 2132 01:37:44,276 --> 01:37:50,849 AND WE FIND OUT IF YOU IMMUNIZE 2133 01:37:50,849 --> 01:37:53,151 MICE -- TRANSFORM CELL, THEY 2134 01:37:53,151 --> 01:37:57,856 WILL KILL -- CYTOTOXIC T-CELL 2135 01:37:57,856 --> 01:38:00,292 BUT ONLY IF THEY WERE FROM THE 2136 01:38:00,292 --> 01:38:01,860 SAME STRAIN WE IMMUNIZED. 2137 01:38:01,860 --> 01:38:10,268 SO B6 MICE ONLY KILL B6 AS -- 2138 01:38:10,268 --> 01:38:21,246 CELL -- CELL,, BALB/C MICE -- T- 2139 01:38:23,248 --> 01:38:24,716 DESCRIBE, BY THE WAY A SURPRISE 2140 01:38:24,716 --> 01:38:26,451 BECAUSE WE ALSO IMMUNIZE MICE 2141 01:38:26,451 --> 01:38:29,087 NOT ONLY -- TRANSFORM CELLS, BUT 2142 01:38:29,087 --> 01:38:35,794 ALSO WITH AL JO ALLOGENEIC -- AD 2143 01:38:35,794 --> 01:38:37,862 REGARDLESS OF WHICH CELL WE USE 2144 01:38:37,862 --> 01:38:42,467 TO IMMUNIZE THE MICE, STILL ALL 2145 01:38:42,467 --> 01:38:47,639 THIS ALLOGENEIC -- SO WE BA BASY 2146 01:38:47,639 --> 01:38:53,311 FOUND THE TUMOR ANTIGEN -- CELL 2147 01:38:53,311 --> 01:38:55,914 ARE CURRENTLY REPRESENTED IN 2148 01:38:55,914 --> 01:38:57,916 VIVO IN SUCCESSION WITH -- 2149 01:38:57,916 --> 01:38:59,951 WHAT'S NOW KNOWN AS ANTIGEN 2150 01:38:59,951 --> 01:39:08,360 CROSS PRESENTATION. 2151 01:39:08,360 --> 01:39:12,197 NOW WE KNOW HOW THE T-CELL 2152 01:39:12,197 --> 01:39:14,799 RECEPTOR WORK AND WE KNOW ALL 2153 01:39:14,799 --> 01:39:17,002 THE MOLECULAR DETAIL OF THESE 2154 01:39:17,002 --> 01:39:19,204 PARTICULAR INTERACTIONS IN TERMS 2155 01:39:19,204 --> 01:39:22,140 OF ACTION. 2156 01:39:22,140 --> 01:39:25,310 BUT IN THE 70s THE T-CELL 2157 01:39:25,310 --> 01:39:27,312 RECEPTOR WAS DESCRIBED BY 2158 01:39:27,312 --> 01:39:30,148 REINHARDT AND MARK DAVIS IN THE 2159 01:39:30,148 --> 01:39:31,549 80s, SO AT THAT TIME WE REALLY 2160 01:39:31,549 --> 01:39:33,184 DIDN'T KNOW WHAT WAS GOING ON 2161 01:39:33,184 --> 01:39:34,819 AND WE HAD A LOT TO DISCUSS WITH 2162 01:39:34,819 --> 01:39:37,689 PETER DOHERTY WHEN HE WAS AT 2163 01:39:37,689 --> 01:39:41,526 WISTAR INTUESDAY AND WE CAME OUT 2164 01:39:41,526 --> 01:39:42,894 WITH THIS CONCEPTUAL PAPER AND 2165 01:39:42,894 --> 01:39:44,362 TRY TO FIGURE OUT WHAT WAS GOING 2166 01:39:44,362 --> 01:39:46,898 ON, AND WE BASICALLY -- DRAWING 2167 01:39:46,898 --> 01:39:50,168 FROM THE PAPER, WE SAID THAT MAY 2168 01:39:50,168 --> 01:39:51,936 BE THAT MH SEA CHANGED THE 2169 01:39:51,936 --> 01:39:55,540 VIRUS, SO THE VIRUS CHANGED MHC 2170 01:39:55,540 --> 01:39:57,509 OR MAYBE THERE'S TWO RECEPTORS, 2171 01:39:57,509 --> 01:39:58,943 MAYBE THERE'S A RECEPTOR THAT 2172 01:39:58,943 --> 01:40:00,812 SEE WHAT IS BETWEEN THE TWO, AND 2173 01:40:00,812 --> 01:40:02,313 ACTUALLY THE FIRST HYPOTHESIS 2174 01:40:02,313 --> 01:40:06,584 THAT WE HAVE, THE POSSIBILITY, 2175 01:40:06,584 --> 01:40:09,454 THE H2MHC WAS MULTIPLIED BY THE 2176 01:40:09,454 --> 01:40:10,755 VIRUS, WE KNOW NOW IT'S 2177 01:40:10,755 --> 01:40:14,926 MULTIPLIED BY THE PEPTIDE, IS 2178 01:40:14,926 --> 01:40:17,695 PROBABLY THE ONE CLOSE WITH 2179 01:40:17,695 --> 01:40:19,664 EVENTUALLY WAS DISCOVERED TO BE 2180 01:40:19,664 --> 01:40:20,532 THE REAL RECOGNITION. 2181 01:40:20,532 --> 01:40:23,935 NOW WHAT ABOUT CROSS 2182 01:40:23,935 --> 01:40:24,669 PRESENTATION? 2183 01:40:24,669 --> 01:40:25,804 EXACTLY AT THE SAME TIME, THE 2184 01:40:25,804 --> 01:40:33,878 SAME MONTH, MAY '76, MIKE -- 2185 01:40:33,878 --> 01:40:34,646 DESCRIBED CROSS PRESENTATION, IN 2186 01:40:34,646 --> 01:40:38,483 THIS CASE IT WAS WORKING WITH 2187 01:40:38,483 --> 01:40:42,420 THE MINOR -- ANTIGEN, AND BEVAN 2188 01:40:42,420 --> 01:40:44,956 AND MANY OTHER REALLY DID ALL 2189 01:40:44,956 --> 01:40:46,691 THE MOLECULAR CHARACTERIZATION 2190 01:40:46,691 --> 01:40:50,095 ANTIGEN CROSS PRESENTATION. 2191 01:40:50,095 --> 01:40:51,629 WHAT WE KNOW NOW IS THAT THE 2192 01:40:51,629 --> 01:40:53,965 ANTIGEN CAN BE CROSS PRESENTED 2193 01:40:53,965 --> 01:40:58,436 BY SPECIALIZED CELL TYPES 2194 01:40:58,436 --> 01:41:01,873 MOSTLY, THIS -- MOSTLY ONE 2195 01:41:01,873 --> 01:41:02,807 SUBSTITUTE DENDRITIC CELL BUT 2196 01:41:02,807 --> 01:41:04,476 AGAIN WHEN WE WERE DOING THE 2197 01:41:04,476 --> 01:41:07,445 STUDY WAS IN '76, THE DENDRITIC 2198 01:41:07,445 --> 01:41:10,715 CELLS WERE ACTUALLY DESCRIBED BY 2199 01:41:10,715 --> 01:41:13,685 STEINMAN IN '73 AS FUNNY LOOKING 2200 01:41:13,685 --> 01:41:16,121 CELLS IN THE LYMPHONODE OF MICE. 2201 01:41:16,121 --> 01:41:19,157 IT WAS NOT ONLY REALLY AFTER 2202 01:41:19,157 --> 01:41:21,226 2004, YES, AFTER THE STUDY I'VE 2203 01:41:21,226 --> 01:41:26,598 BEEN DISCUSSING NOW, BEVAN'S 2204 01:41:26,598 --> 01:41:30,201 STUDY, THAT -- EVENTUALLY 2205 01:41:30,201 --> 01:41:33,271 IDENTIFIED THE DENDRITIC CELL AS 2206 01:41:33,271 --> 01:41:37,542 THE ANTIGEN PRESENTING CELL IN 2207 01:41:37,542 --> 01:41:38,743 CULTURE AND LYMPHOCYTE PRESENT 2208 01:41:38,743 --> 01:41:39,477 BEING IN GENERAL. 2209 01:41:39,477 --> 01:41:41,713 AND AT THAT TIME OBVIOUSLY WE 2210 01:41:41,713 --> 01:41:46,985 COULD NOT ATTRIBUTE -- NOR BEVAN 2211 01:41:46,985 --> 01:41:48,686 TO DENDRITIC CELL AND WHAT WE 2212 01:41:48,686 --> 01:41:52,090 WERE REALLY THINKING WAS WITH -- 2213 01:41:52,090 --> 01:41:54,425 IT WAS -- A MAJOR ANTIGEN 2214 01:41:54,425 --> 01:42:00,131 PRESENTING CELL WERE A MACROPHA, 2215 01:42:00,131 --> 01:42:01,966 THE TUMOR CELL IN VIVO AND THEN 2216 01:42:01,966 --> 01:42:06,037 PRESENT ON THEIR H2 TO THE 2217 01:42:06,037 --> 01:42:06,938 IMMUNE SYSTEM OF THE HOST. 2218 01:42:06,938 --> 01:42:13,278 I'D LIKE TO MENTION EMIL UNANUE, 2219 01:42:13,278 --> 01:42:14,479 ACTUALLY PASSED AWAY A SHORT 2220 01:42:14,479 --> 01:42:16,681 TIME AGO, AND -- WAS THE ONE 2221 01:42:16,681 --> 01:42:18,216 THAT NOMINATED ME FOR THE 2222 01:42:18,216 --> 01:42:23,354 ACADEMY. 2223 01:42:23,354 --> 01:42:27,525 SO THE OTHER WHEN WE RECOGNIZE 2224 01:42:27,525 --> 01:42:32,664 THAT THE HUMAN -- HLA CLASS I, 2225 01:42:32,664 --> 01:42:36,067 WE THOUGHT THAT MAYBE IF WE USE 2226 01:42:36,067 --> 01:42:38,670 ANTIBODY AGAINST HLA CLASS 1, 2227 01:42:38,670 --> 01:42:41,306 REMEMBER THERE WERE NO 2228 01:42:41,306 --> 01:42:42,407 MONOCLONAL ANTIBODY AT THAT 2229 01:42:42,407 --> 01:42:46,411 TIME, THERE WERE SE SERA FROM A 2230 01:42:46,411 --> 01:42:47,478 WOMAN, MAYBE WE CAN BLOCK THE 2231 01:42:47,478 --> 01:42:48,012 KILLING. 2232 01:42:48,012 --> 01:42:50,515 IF WE'RE USING TO TRY TO 2233 01:42:50,515 --> 01:42:51,849 COMPLETELY BLOCK, WE DID NOT 2234 01:42:51,849 --> 01:42:53,818 REALLY BLOCK THE KILLING BUT WE 2235 01:42:53,818 --> 01:42:56,254 GOT MORE KILLING AND THE REASON 2236 01:42:56,254 --> 01:42:58,623 WAS THAT WE'RE IN OUR CULTURE, 2237 01:42:58,623 --> 01:43:00,091 NATURAL KILLER CELLS THAT WERE 2238 01:43:00,091 --> 01:43:03,261 ABLE TO DO ANTIBODY 2239 01:43:03,261 --> 01:43:04,462 CYTOTOXICITY, REALLY BROUGHT FOR 2240 01:43:04,462 --> 01:43:07,565 SEVERAL YEARS IN MY GROUP TO 2241 01:43:07,565 --> 01:43:11,069 STUDY MANY OF THE BIOLOGY OF 2242 01:43:11,069 --> 01:43:12,270 THIS INNATE LYMPHOCYTE OF 2243 01:43:12,270 --> 01:43:15,240 NATURAL KILLER CELL. 2244 01:43:15,240 --> 01:43:18,943 SO WITH -- WE DESCRIBED FOR 2245 01:43:18,943 --> 01:43:21,913 EXAMPLE THE RECEPTOR RECOGNIZING 2246 01:43:21,913 --> 01:43:23,881 IMMUNOGLOBULIN ON THE NATIONAL 2247 01:43:23,881 --> 01:43:26,084 KILLER CELL, AND WE DISCOVER 2248 01:43:26,084 --> 01:43:29,354 THAT IT WAS A MAJOR ACTIVATOR 2249 01:43:29,354 --> 01:43:30,555 FOR NATURAL KILLER CELL 2250 01:43:30,555 --> 01:43:34,959 FUNCTION, BUT WE ALSO FIND OUT 2251 01:43:34,959 --> 01:43:37,462 THAT THE NK CELL THAT WERE KNOWN 2252 01:43:37,462 --> 01:43:38,663 TO KILL TUMOR CELL OR TEU NOR 2253 01:43:38,663 --> 01:43:41,633 CELL LINE, WERE ALSO VERY EE 2254 01:43:41,633 --> 01:43:42,834 EFFICIENT IN KILLING VIRUS 2255 01:43:42,834 --> 01:43:46,704 INFECTED CELL BUT NOT THE SAME 2256 01:43:46,704 --> 01:43:47,672 FIBROBLAST -- IN THIS CASE WERE 2257 01:43:47,672 --> 01:43:48,973 NOT INFECTED BY VIRUS. 2258 01:43:48,973 --> 01:43:54,445 AND WHEN WE LOOK AT WHAT WAS 2259 01:43:54,445 --> 01:43:55,780 HAPPENING THE AFFECTED CELL, WE 2260 01:43:55,780 --> 01:43:58,082 FOUND OUT THAT PRODUCTION OF 2261 01:43:58,082 --> 01:43:59,817 TYPE 1 INTERFERON WAS NEEDED TO 2262 01:43:59,817 --> 01:44:03,454 ACTIVATE NK CELLS IN VIRUS 2263 01:44:03,454 --> 01:44:03,988 INFECTED CELLS. 2264 01:44:03,988 --> 01:44:06,224 BUT WHEN WE TRY TO IDENTIFY 2265 01:44:06,224 --> 01:44:08,159 WHICH CELL IN THE CULTURE WAS 2266 01:44:08,159 --> 01:44:09,961 PRODUCING INTERFERON, THAT WAS 2267 01:44:09,961 --> 01:44:12,930 NOT NK CELLS, THEY WERE NOT THE 2268 01:44:12,930 --> 01:44:14,666 VIRUS INFECTED CELL BUT ACTUALLY 2269 01:44:14,666 --> 01:44:18,136 A VERY RARE CELL TYPE IN HUMAN 2270 01:44:18,136 --> 01:44:21,873 PERIPHERAL BLOOD THAT WE DEFINE 2271 01:44:21,873 --> 01:44:29,847 AS THE -- APOLOGIZE, THIS SEEMS 2272 01:44:29,847 --> 01:44:31,082 TO BE FROZEN. 2273 01:44:31,082 --> 01:44:31,282 OKAY. 2274 01:44:31,282 --> 01:44:35,453 LET'S GO BACK. 2275 01:44:35,453 --> 01:44:43,094 THAT WE RECOGNIZE -- THAT WE 2276 01:44:43,094 --> 01:44:44,962 DEFINE NATURAL TYPE 1 INTERFERON 2277 01:44:44,962 --> 01:44:46,831 PRODUCING CELL AND CELL TYPE NOW 2278 01:44:46,831 --> 01:44:52,737 IS VERY WELL STUDIED -- 2279 01:44:52,737 --> 01:44:53,404 DENDRITIC CELL. 2280 01:44:53,404 --> 01:44:55,373 WE FOUND THESE CELLS WERE THE 2281 01:44:55,373 --> 01:44:59,877 MOST EFFICIENT PRODUCER OF TYPE 2282 01:44:59,877 --> 01:45:00,511 1 IN PERIPHERAL BLOOD. 2283 01:45:00,511 --> 01:45:03,181 THERE WERE SOME -- WITH 2284 01:45:03,181 --> 01:45:04,515 DENDRITIC CELL BUT ACTUALLY 2285 01:45:04,515 --> 01:45:05,917 QUITE DIFFERENT, HERE I SHOW THE 2286 01:45:05,917 --> 01:45:07,785 MORPHOLOGY OF HUMAN DENDRITIC 2287 01:45:07,785 --> 01:45:10,088 CELL IN THE BLOOD VERSUS THE 2288 01:45:10,088 --> 01:45:12,590 INTERFERON PRODUCING CELLS. 2289 01:45:12,590 --> 01:45:15,126 AND WE STUDIED THAT FOR A FEW 2290 01:45:15,126 --> 01:45:17,228 YEARS, BUT WHAT WAS SURPRISING 2291 01:45:17,228 --> 01:45:21,165 WAS UNTIL ABOUT 20 YEARS LATER, 2292 01:45:21,165 --> 01:45:22,266 IMMUNOLOGISTS IN MOUSE WERE 2293 01:45:22,266 --> 01:45:23,701 CLAIMING THAT THIS CELL DID NOT 2294 01:45:23,701 --> 01:45:24,669 EXIST IN THE MOUSE, THEY WERE 2295 01:45:24,669 --> 01:45:25,536 ONLY PRESENT IN HUMAN. 2296 01:45:25,536 --> 01:45:29,841 SO WE WENT TO TRY WHEN I WAS IN 2297 01:45:29,841 --> 01:45:31,142 ROSAN TO TRY TO IDENTIFY THE 2298 01:45:31,142 --> 01:45:32,710 CELL AND WE FOUND OUT ACTUALLY 2299 01:45:32,710 --> 01:45:33,511 THEY'RE VERY SIMILAR IN THE 2300 01:45:33,511 --> 01:45:34,679 MOUSE AS IN HUMAN. 2301 01:45:34,679 --> 01:45:36,848 THEY ARE MOUSE TYPE 1 PRODUCING 2302 01:45:36,848 --> 01:45:42,353 CELL AND THIS WAS DEFINED 2303 01:45:42,353 --> 01:45:45,189 PLASMA -- MORPHOLOGY AND THEY'RE 2304 01:45:45,189 --> 01:45:46,090 NOW -- DENDRITIC CELL. 2305 01:45:46,090 --> 01:45:47,625 WHEN I MENTIONED THAT THIS 2306 01:45:47,625 --> 01:45:50,695 REALLY BROUGHT ME TO COLLABORATE 2307 01:45:50,695 --> 01:45:52,330 FOR MANY, MANY YEARS WAS A VERY 2308 01:45:52,330 --> 01:45:54,532 CLOSE FRIEND OF MINE, CHRISTINE 2309 01:45:54,532 --> 01:45:58,469 BIRON, THAT WE DID THE MAJOR 2310 01:45:58,469 --> 01:46:00,438 PLAYER IN THE NK CELL FIELD AND 2311 01:46:00,438 --> 01:46:01,706 VIRUS INFECTION, AND WE ARE 2312 01:46:01,706 --> 01:46:03,441 HAPPY TO COLLABORATE IN SOME OF 2313 01:46:03,441 --> 01:46:07,145 THIS STUDY. 2314 01:46:07,145 --> 01:46:10,515 NOW, INTERFERON CAN INDUCE -- IN 2315 01:46:10,515 --> 01:46:13,184 CASE -- BECAME CYTOTOXIC -- WE 2316 01:46:13,184 --> 01:46:15,586 FOUND A FEW YEARS LATER THAT 2317 01:46:15,586 --> 01:46:21,192 OTHER CYTOKINE LIKE INTERLEUKIN 2318 01:46:21,192 --> 01:46:22,727 2 OR STIMULATION THROUGH THE -- 2319 01:46:22,727 --> 01:46:24,695 RECEPTOR WERE NOT ONLY -- TO 2320 01:46:24,695 --> 01:46:26,264 BECOME MORE CYTOTOXIC, ALSO USE 2321 01:46:26,264 --> 01:46:29,967 THEM TO PROLIFERATE BUT MOST 2322 01:46:29,967 --> 01:46:31,169 IMPORTANT WAS INDUCE THEM TO 2323 01:46:31,169 --> 01:46:33,638 PRODUCE A CYTOKINE. 2324 01:46:33,638 --> 01:46:35,339 THE MAJOR ONE IS INTERFERON 2325 01:46:35,339 --> 01:46:37,141 GAMMA BUT ALSO THE CYTOKINE 2326 01:46:37,141 --> 01:46:37,542 LISTED HERE. 2327 01:46:37,542 --> 01:46:44,315 NOW THIS FINDING THAT -- NK CELL 2328 01:46:44,315 --> 01:46:47,051 AND -- T-CELL -- WITHOUT 2329 01:46:47,051 --> 01:46:50,888 SENSITIZATION WAS REALLY THE 2330 01:46:50,888 --> 01:46:54,058 FIRST IDENTIFICATION OF NK CELL 2331 01:46:54,058 --> 01:46:56,160 IS AN EXAMPLE OF -- TYPE I 2332 01:46:56,160 --> 01:46:59,230 SIMILAR TO NK CELL, TYPE 2 AND 2333 01:46:59,230 --> 01:46:59,497 TYPE 3. 2334 01:46:59,497 --> 01:47:00,665 BUT THE OTHER THING THAT WE WERE 2335 01:47:00,665 --> 01:47:02,500 VERY INTERESTED IN AT THAT TIME 2336 01:47:02,500 --> 01:47:05,236 WAS THE FACT NK CELLS WERE KNOWN 2337 01:47:05,236 --> 01:47:09,640 TO BE ABLE TO REGULATE 2338 01:47:09,640 --> 01:47:12,910 HEMOPOIESIS, INTERFERON GAMMA -- 2339 01:47:12,910 --> 01:47:19,717 MACROPHAGE AND THE -- CYTOKINE 2340 01:47:19,717 --> 01:47:23,120 COULD INDUCE -- OF -- MAJOR 2341 01:47:23,120 --> 01:47:27,391 ROLE -- BUT ALSO TWO OTHER 2342 01:47:27,391 --> 01:47:28,626 CYTOKINE, WE ARE ALL WORKING 2343 01:47:28,626 --> 01:47:30,695 TOGETHER WITH INTERFERON GAMMA 2344 01:47:30,695 --> 01:47:35,600 IN HAVING THIS EFFECT OF MYELOID 2345 01:47:35,600 --> 01:47:39,337 DIFFERENTIATION AND INHIBITION 2346 01:47:39,337 --> 01:47:42,073 OF HEMATOPOIESIS. 2347 01:47:42,073 --> 01:47:44,942 SO THESE ARE THE CYTOKINES THAT 2348 01:47:44,942 --> 01:47:50,515 ARE THOUGHT TO BE GENE CLONE. 2349 01:47:50,515 --> 01:47:55,119 WE COULD NOT GET -- FOR TOXINS 2350 01:47:55,119 --> 01:47:58,556 SO WE DECIDE WE HAD TO PURIFY 2351 01:47:58,556 --> 01:47:58,856 LYMPHOTOXIN. 2352 01:47:58,856 --> 01:48:03,027 AND THE GOOD SOURCE OF 2353 01:48:03,027 --> 01:48:05,630 LYMPHOTOXIN IS FROM EBV 2354 01:48:05,630 --> 01:48:07,598 TRANSFORMED HUMAN B CELL LINE. 2355 01:48:07,598 --> 01:48:10,935 SO WE STARTED PURIFYING FROM ONE 2356 01:48:10,935 --> 01:48:16,674 OF THE CELL LINE, AND WHY WE ARE 2357 01:48:16,674 --> 01:48:21,212 DOING THAT AN WE'RE TESTING IN 2358 01:48:21,212 --> 01:48:23,748 FRACTIONATION SUPER NATANT -- 2359 01:48:23,748 --> 01:48:26,017 BECAUSE WE KNEW THAT THE B CELL 2360 01:48:26,017 --> 01:48:28,786 LINE ALSO ACTIVATED AND CAN 2361 01:48:28,786 --> 01:48:30,888 INTRODUCE INTERFERON PRODUCTION 2362 01:48:30,888 --> 01:48:32,189 WITH -- IF WE'RE SEEING THE 2363 01:48:32,189 --> 01:48:35,059 SUPER NATANT OR SOME ABILITY TO 2364 01:48:35,059 --> 01:48:37,562 STIMULATE INTERFERON GAMMA OR 2365 01:48:37,562 --> 01:48:38,563 ACTIVATE NK CELLS. 2366 01:48:38,563 --> 01:48:41,999 THIS IS REALLY THE FIRST TWO 2367 01:48:41,999 --> 01:48:43,501 COLUMBUS WE DID. 2368 01:48:43,501 --> 01:48:49,073 THE FIRST COLUMN WAS -- WE'VE 2369 01:48:49,073 --> 01:48:51,909 FOUND A BROAD PEAK AND ALSO 2370 01:48:51,909 --> 01:48:54,545 INTERFATHER ON GAMMA PRODUCTIONS 2371 01:48:54,545 --> 01:48:58,249 STIMULATING NK CELL, BUT THEN 2372 01:48:58,249 --> 01:49:00,451 WHEN -- PEER FIED BY ION 2373 01:49:00,451 --> 01:49:03,454 EXCHANGE CHROME MAAING TO FEE, 2374 01:49:03,454 --> 01:49:05,122 WE COULD COMPLETELY SEPARATE 2375 01:49:05,122 --> 01:49:07,592 FROM THE ABILITY TO STIMULATE N 2376 01:49:07,592 --> 01:49:11,095 KSM CELL TO PRODUCE INTERFERON 2377 01:49:11,095 --> 01:49:12,630 GAMMA. 2378 01:49:12,630 --> 01:49:16,567 SO WE DEFINE THIS NEW ACTIVITY 2379 01:49:16,567 --> 01:49:19,103 AS A NATURAL KILLER CELL 2380 01:49:19,103 --> 01:49:23,074 STIMULATORY FACTOR THAT WAS -- 2381 01:49:23,074 --> 01:49:26,310 ABILITY TO -- INDUCE INTERFERON 2382 01:49:26,310 --> 01:49:27,745 GAMMA PRODUCTION. 2383 01:49:27,745 --> 01:49:33,651 AND WHEN WE BEAUTIFY ESPECIALLY 2384 01:49:33,651 --> 01:49:36,954 THE WORK BY THAT TIME, WE FIND 2385 01:49:36,954 --> 01:49:41,559 OUT THERE IS A VAN AROUND 70 -- 2386 01:49:41,559 --> 01:49:43,060 BUT WHETHER THIS WAS REDUCED, IT 2387 01:49:43,060 --> 01:49:44,929 WAS ACTUALLY FORMED BY TWO 2388 01:49:44,929 --> 01:49:49,200 CHAINS, ONE OF 40,000, P40, ONE 2389 01:49:49,200 --> 01:49:52,103 OF THE 35,000 OR P35. 2390 01:49:52,103 --> 01:49:54,438 SO THIS WAS ACTUALLY NOT A 2391 01:49:54,438 --> 01:49:58,576 SINGLE CHAIN CYTOKINE, WAS 2392 01:49:58,576 --> 01:50:00,211 ACTUALLY -- IN WHICH THE 2393 01:50:00,211 --> 01:50:00,878 ACTIVITY WAS FOUND. 2394 01:50:00,878 --> 01:50:02,847 SO THAT WAS VERY SURPRISING 2395 01:50:02,847 --> 01:50:06,150 BECAUSE ALL THAT CYTOKINE AT 2396 01:50:06,150 --> 01:50:07,451 THAT TIME WERE KNOWN TO BE 2397 01:50:07,451 --> 01:50:08,686 SINGLE CHAIN, BUT THEN WE 2398 01:50:08,686 --> 01:50:09,687 ACTUALLY FOUND OUT MAYBE WE 2399 01:50:09,687 --> 01:50:12,723 SHOULD NOT BE SURPRISED BECAUSE 2400 01:50:12,723 --> 01:50:17,461 IL-1, IL2, IL-8, IL10 ARE NOT 2401 01:50:17,461 --> 01:50:24,669 ONLY SITE KIN, BUT THE FIRST 2402 01:50:24,669 --> 01:50:29,874 AIRPLANE, DUAL ENGINE, WAS. 2403 01:50:29,874 --> 01:50:33,678 IL2. 2404 01:50:33,678 --> 01:50:37,081 FOUND STRUCTURE -- WHEN 2405 01:50:37,081 --> 01:50:38,716 HETERODIMER FORM BY IL-6 AND 2406 01:50:38,716 --> 01:50:42,687 FORM OF IL-6 RECEPTOR. 2407 01:50:42,687 --> 01:50:46,724 SO THE IL12 WAS STARTED BY US 2408 01:50:46,724 --> 01:50:48,959 AND MANY OTHER, THE MOLECULES 2409 01:50:48,959 --> 01:50:51,896 WERE CHARACTERIZED, THE RECEPTOR 2410 01:50:51,896 --> 01:51:01,072 WAS CHARACTERIZED, FROM JACK -- 2411 01:51:01,072 --> 01:51:05,476 IF YOU'RE LISTENING TO TESTIMONY 2412 01:51:05,476 --> 01:51:06,777 VCH -- VERY FAMOUS MOLECULES, 2413 01:51:06,777 --> 01:51:08,345 AND THE SIGNALS PROVES THAT 2414 01:51:08,345 --> 01:51:11,982 SIGNALING MOST AT FOUR WOULD 2415 01:51:11,982 --> 01:51:14,151 INDUCE INTERFERON GAMMA AND 2416 01:51:14,151 --> 01:51:15,920 OTHER ACTIVATED MONIKERS. 2417 01:51:15,920 --> 01:51:18,456 SO INTERLEUKIN 12 ENDED UP TO BE 2418 01:51:18,456 --> 01:51:20,324 A CYTOKINE THAT PROVIDES THE 2419 01:51:20,324 --> 01:51:22,159 CRITICAL LINK BETWEEN INNATE 2420 01:51:22,159 --> 01:51:24,395 ADAPTIVE IMMUNITY THAT'S 2421 01:51:24,395 --> 01:51:26,697 PRODUCED BY -- MOSTLY BY MYELOID 2422 01:51:26,697 --> 01:51:28,032 CELL, DENDRITIC CELL IN RESPONSE 2423 01:51:28,032 --> 01:51:30,334 TO MANY STIMULI, INCLUDING 2424 01:51:30,334 --> 01:51:32,770 INFECTION, BACTERIAL AND VIRAL, 2425 01:51:32,770 --> 01:51:35,272 AND ACTIVATING ON NK CELL AND 2426 01:51:35,272 --> 01:51:38,876 T-CELL. 2427 01:51:38,876 --> 01:51:42,646 AND IN 1993, WE WORK WITH 2428 01:51:42,646 --> 01:51:43,714 SERGIO -- COULD ACTUALLY SEE 2429 01:51:43,714 --> 01:51:45,616 THAT IL12 IS THE MAJOR MOLECULES 2430 01:51:45,616 --> 01:51:48,552 THAT IS INVOLVED IN THE 2431 01:51:48,552 --> 01:51:49,653 PRIORITIZATION OF THE FUNCTIONAL 2432 01:51:49,653 --> 01:51:53,724 SUBSET OF T-CELL, THE TH1 2433 01:51:53,724 --> 01:51:56,160 T-CELL, THE TL -- 1 THAT ARE THE 2434 01:51:56,160 --> 01:51:57,895 T-CELL THAT PRODUCE INTERFERON 2435 01:51:57,895 --> 01:51:59,196 GAMMA. 2436 01:51:59,196 --> 01:52:01,432 AND AGAIN, LIKE FOR CROSS 2437 01:52:01,432 --> 01:52:03,067 PRESENTATION, EXACTLY IN THE 2438 01:52:03,067 --> 01:52:07,438 SAME MONTH, IN 1993, THEY 2439 01:52:07,438 --> 01:52:08,572 PUBLISHED EXACTLY THE SAME 2440 01:52:08,572 --> 01:52:12,843 RESULT IN THE MOUSE. 2441 01:52:12,843 --> 01:52:16,781 AT THAT TIME OBVIOUSLY -- WAS A 2442 01:52:16,781 --> 01:52:18,649 MAJOR COMPETITOR OF US AND NOW 2443 01:52:18,649 --> 01:52:20,651 BECAME A WONDERFUL FRIEND AND 2444 01:52:20,651 --> 01:52:21,719 MANY COLLABORATIONS WE'VE DONE. 2445 01:52:21,719 --> 01:52:26,123 ONE I'D LIKE TO MENTION ACTUALLY 2446 01:52:26,123 --> 01:52:29,093 ANNE WAS ELECTED TO THE ACADEMY 2447 01:52:29,093 --> 01:52:31,395 OF SCIENCE AS INTERNATIONAL 2448 01:52:31,395 --> 01:52:32,730 ACADEMY MEMBER. 2449 01:52:32,730 --> 01:52:35,032 SO WHEN WE STARTED LOOKING 2450 01:52:35,032 --> 01:52:36,767 BETTER AT THE INTERACTION OF THE 2451 01:52:36,767 --> 01:52:41,605 DIFFERENT CHAIN WITH DELTA, WE 2452 01:52:41,605 --> 01:52:43,040 ACTUALLY START SEEING WHERE 2453 01:52:43,040 --> 01:52:44,475 OTHER CHAINS THAT WERE 2454 01:52:44,475 --> 01:52:45,676 INTERACTING WITH BEFORE, 2455 01:52:45,676 --> 01:52:47,845 PARTICULARLY ONE IS CALLED P19, 2456 01:52:47,845 --> 01:52:51,048 THEN NOW WE KNOW THIS FORM 2457 01:52:51,048 --> 01:52:53,684 ANOTHER CYTOKINE THAT WAS NOT 2458 01:52:53,684 --> 01:52:56,687 REALLY THE WORK IF MY LAB, IT 2459 01:52:56,687 --> 01:53:01,058 WAS MOSTLY THE WORK AT THAT 2460 01:53:01,058 --> 01:53:04,228 TIME -- FERNANDO, ROB AND -- 2461 01:53:04,228 --> 01:53:08,833 THAT REALLY DESCRIBE MANY IL23 2462 01:53:08,833 --> 01:53:10,234 AND OTHER CYTOKINE OF THIS 2463 01:53:10,234 --> 01:53:12,002 FAMILY AND THIS REALLY WHAT HAS 2464 01:53:12,002 --> 01:53:16,841 BECOME NOW AS A GROWING FAMILY 2465 01:53:16,841 --> 01:53:21,879 OF HETERODIMERIC CYTOKINE, AND 2466 01:53:21,879 --> 01:53:23,314 IL23 AT THE BEGINNING WITH SOME 2467 01:53:23,314 --> 01:53:25,282 OF THE ACTIVITY IL23 WERE 2468 01:53:25,282 --> 01:53:28,285 ASSIGNED TO US TO HAVE BUT WHAT 2469 01:53:28,285 --> 01:53:30,487 ACTUALLY DUE TO THE ABILITY OF 2470 01:53:30,487 --> 01:53:35,192 I23 NOT TO -- AND THEREFORE 2471 01:53:35,192 --> 01:53:37,094 IL-23 IS ACTUALLY A MAJOR ROLE 2472 01:53:37,094 --> 01:53:40,898 IN AUTOIMMUNITY. 2473 01:53:40,898 --> 01:53:43,300 THE WORK ON IL12 REALLY STARTED 2474 01:53:43,300 --> 01:53:45,336 WHAT WAS VERY CLOSE FOR ALMOST 2475 01:53:45,336 --> 01:53:46,637 10 YEAR COLLABORATION BETWEEN 2476 01:53:46,637 --> 01:53:49,273 PHILADELPHIA AND BETHESDA. 2477 01:53:49,273 --> 01:53:56,647 WITH THE LAB OF SHERRA AS WELL 2478 01:53:56,647 --> 01:53:59,917 AS ALUMNI WILL LIKE PHIL SCOTT 2479 01:53:59,917 --> 01:54:04,188 THAT WAS ALREADY IN PHILADELPHIA 2480 01:54:04,188 --> 01:54:10,294 DEFINITELY YA, IF THERE'S -- 2481 01:54:10,294 --> 01:54:12,529 COME TO BETHESDA FOR LAB MEETING 2482 01:54:12,529 --> 01:54:14,198 AND WITH ALL THESE 2483 01:54:14,198 --> 01:54:16,467 COLLABORATIONS WE WORK ON 2484 01:54:16,467 --> 01:54:19,570 EFFECTIVE IL20, IN CANCER, IN 2485 01:54:19,570 --> 01:54:21,205 INFECTION, AND IN AUTOIMMUNITY. 2486 01:54:21,205 --> 01:54:26,277 AND IT WAS REALLY A WOU WONDERFL 2487 01:54:26,277 --> 01:54:28,345 TIME OF COLLABORATIVE WORK ON 2488 01:54:28,345 --> 01:54:30,881 ALL THESE DIFFERENT ASPECTS. 2489 01:54:30,881 --> 01:54:33,117 AND I MENTIONED TO YOU THAT IL12 2490 01:54:33,117 --> 01:54:37,388 IS INDUCED BY MOSTLY MYELOID 2491 01:54:37,388 --> 01:54:40,090 CELL AND DENDRITIC CELL 2492 01:54:40,090 --> 01:54:41,258 STIMULATION WITH PATHOGEN OR 2493 01:54:41,258 --> 01:54:46,196 WITH BACTERIA OR WITH VIRUS. 2494 01:54:46,196 --> 01:54:47,932 AND THAT REALLY BROUGHT THE 2495 01:54:47,932 --> 01:54:49,166 THINKING BACK OF WHAT IS THE 2496 01:54:49,166 --> 01:54:52,436 WORK OF WILLIAM COLEY WITH THE 2497 01:54:52,436 --> 01:54:56,173 COLEY TOXIN WHICH WAS ABLE TO 2498 01:54:56,173 --> 01:54:58,042 TREAT OSTEOSARCOMA WITH A 2499 01:54:58,042 --> 01:55:02,746 MIXTURE OF BACK YEAR YA, STRECT 2500 01:55:02,746 --> 01:55:07,217 COCCUS AND -- AND INDUCE 2501 01:55:07,217 --> 01:55:10,654 INFLAMMATION AND -- REME REMISSN 2502 01:55:10,654 --> 01:55:11,422 SOME PATIENT AND WHAT WAS 2503 01:55:11,422 --> 01:55:13,891 CONSIDERED TO BE PROBABLY THE 2504 01:55:13,891 --> 01:55:16,060 FIRST IN THE LATE 19TH CENTURY, 2505 01:55:16,060 --> 01:55:18,696 THE FIRST FORM OF IMMUNOTHERAPY. 2506 01:55:18,696 --> 01:55:25,402 AND AS I MENTIONED, INDEED A 2507 01:55:25,402 --> 01:55:28,872 COLEY TOXIN -- ACTIVATE -- THAT 2508 01:55:28,872 --> 01:55:31,642 KILLED THE TUMOR AND OUR WORK AN 2509 01:55:31,642 --> 01:55:38,248 MANY OTHER LAB -- IN TUMOR 2510 01:55:38,248 --> 01:55:38,682 KILLING. 2511 01:55:38,682 --> 01:55:43,387 THAT'S REALLY BROAD TO STEVE 2512 01:55:43,387 --> 01:55:47,524 BALDWIN, QUITE POPULAR IN THE 2513 01:55:47,524 --> 01:55:48,892 IMMUNOTHERAPY FIELD. 2514 01:55:48,892 --> 01:55:52,963 IN WHICH IL12 WAS DEFINED AS THE 2515 01:55:52,963 --> 01:55:57,234 NEXT GREAT MAGIC BULLET. 2516 01:55:57,234 --> 01:56:00,537 THAT TIME OF THIS INTERACTION 2517 01:56:00,537 --> 01:56:03,507 WITH -- IN THE TUMOR, TUMOR 2518 01:56:03,507 --> 01:56:04,608 IMMUNOTHERAPY, REALLY BROUGHT ME 2519 01:56:04,608 --> 01:56:09,346 VERY CLOSE TO THE CANCER 2520 01:56:09,346 --> 01:56:10,547 RESEARCH INSTITUTE IN WHICH I'M 2521 01:56:10,547 --> 01:56:12,316 STILL INVOLVED AFTER MANY YEARS, 2522 01:56:12,316 --> 01:56:13,784 AND THE CANCER RESEARCH 2523 01:56:13,784 --> 01:56:19,223 INSTITUTE, YOU MAY KNOW, WAS 2524 01:56:19,223 --> 01:56:24,728 FOUNDED BY DR. HELENE COLEY 2525 01:56:24,728 --> 01:56:26,497 NAUTS, AND THAT REALLY BROUGHT 2526 01:56:26,497 --> 01:56:35,439 MANY DISCUSSIONS -- THAT WAS -- 2527 01:56:35,439 --> 01:56:36,440 OF THE CANCER RESEARCH 2528 01:56:36,440 --> 01:56:37,241 INSTITUTE. 2529 01:56:37,241 --> 01:56:39,676 SO THIS WAS THE NEXT GREAT MAGIC 2530 01:56:39,676 --> 01:56:44,815 BULLET, SO DID WE CURE CANCER? 2531 01:56:44,815 --> 01:56:46,216 IT'S GREAT IN THE MOUSE, IT'S 2532 01:56:46,216 --> 01:56:51,622 CONSIDERED THE MOST POTENT 2533 01:56:51,622 --> 01:56:52,923 CYTOKINE -- SO TOXIC THAT IT WAS 2534 01:56:52,923 --> 01:56:54,591 REALLY NEVER SUCCESSFULLY USED 2535 01:56:54,591 --> 01:56:57,561 IN THE CLINICS, AND THERE WERE 2536 01:56:57,561 --> 01:56:59,863 ACTUALLY MAJOR TOXIC EFFECT 2537 01:56:59,863 --> 01:57:01,398 DURING THE CLINICAL TRIAL. 2538 01:57:01,398 --> 01:57:04,668 THERE IS STILL WORK, THERE'S 2539 01:57:04,668 --> 01:57:07,137 STILL ATTEMPT TO USE IN A MORE 2540 01:57:07,137 --> 01:57:10,407 LOCALIZED TARGET WAY TO AVOID 2541 01:57:10,407 --> 01:57:10,674 TOXICITY. 2542 01:57:10,674 --> 01:57:12,643 WE DID SOME WORK WITH STEVE 2543 01:57:12,643 --> 01:57:17,314 ROSENBERG ON THAT AND ON CAR-T 2544 01:57:17,314 --> 01:57:18,615 CELLS AND SOME PEOPLE ARE STILL 2545 01:57:18,615 --> 01:57:20,284 WORKING ON THAT, AND FOR 2546 01:57:20,284 --> 01:57:22,619 EXAMPLE, ROSA KAPLAN AT NCI IS 2547 01:57:22,619 --> 01:57:25,589 REALLY GETTING VERY NICE WORK 2548 01:57:25,589 --> 01:57:32,262 WITH TRANSFECTING MYELOID CELLS 2549 01:57:32,262 --> 01:57:36,400 WITH -- AVENUES IN THERAPY. 2550 01:57:36,400 --> 01:57:40,971 SO THE ANTIBODY AGAINST THE 2551 01:57:40,971 --> 01:57:45,209 P4010IL12 WOULD BLOCK -- WAS 2552 01:57:45,209 --> 01:57:48,245 APPROVED BY FDA FOR VIE CYST, 2553 01:57:48,245 --> 01:57:55,352 ARTHRITIS AND THE DRUG -- IS 2554 01:57:55,352 --> 01:57:57,087 ACTUALLY SELLING MORE THAN 2555 01:57:57,087 --> 01:57:58,255 $10 BILLION PER YEAR. 2556 01:57:58,255 --> 01:58:02,192 AND I DON'T KNOW IF I'M PROUD OR 2557 01:58:02,192 --> 01:58:04,595 ASHAMED OF THAT BUT IT'S 2558 01:58:04,595 --> 01:58:07,698 ACTUALLY ONE OF THE 10 DRUGS BY 2559 01:58:07,698 --> 01:58:11,502 THE ADMINISTRATION TARGETED FOR 2560 01:58:11,502 --> 01:58:12,369 PRICE REDUCTION. 2561 01:58:12,369 --> 01:58:13,470 PROBABLY THIS DRUG IS NOT GOING 2562 01:58:13,470 --> 01:58:15,339 TO BE AROUND FOR LONG BECAUSE 2563 01:58:15,339 --> 01:58:17,674 NOW IT'S A SPECIFIC ANTIBODY 2564 01:58:17,674 --> 01:58:22,379 AGAINST IL20 THEY BUT MUCH 2565 01:58:22,379 --> 01:58:24,581 MORE -- AN MOST LIKELY THEY WILL 2566 01:58:24,581 --> 01:58:28,552 REPLACE THE LESS SPECIFIC 2567 01:58:28,552 --> 01:58:31,755 ANTI -- SO IN 2004, I WANT TO 2568 01:58:31,755 --> 01:58:33,924 LEAVE MY ADVENTURE IN BIG PHARMA 2569 01:58:33,924 --> 01:58:36,393 WHEN I WAS WORKING FOR 2570 01:58:36,393 --> 01:58:37,461 SCHERING-PLOUGH AND I WAS HAPPY 2571 01:58:37,461 --> 01:58:38,695 TO BE ABLE TO DO BASIC SAY ENS 2572 01:58:38,695 --> 01:58:45,836 THERI SCIENCETHERE BUT YOU DON'G 2573 01:58:45,836 --> 01:58:48,005 DOING BASIC SCIENCE IN A BIG 2574 01:58:48,005 --> 01:58:51,675 PHARMA COMPANY. 2575 01:58:51,675 --> 01:58:52,776 TWO PERSON I REALLY NEED TO 2576 01:58:52,776 --> 01:58:54,945 THANK FOR THAT, ONE IS MY OLD 2577 01:58:54,945 --> 01:58:58,282 FRIEND ALAN SHER THAT BROUGHT ME 2578 01:58:58,282 --> 01:58:59,816 FOR SIX MONTH SABBATICAL THAT 2579 01:58:59,816 --> 01:59:02,119 LASTING TWO YEARS IN HIS LAB, 2580 01:59:02,119 --> 01:59:07,324 AND KATHY ZOON AND TONY FAUCI 2581 01:59:07,324 --> 01:59:08,425 REALLY SUPPORTED THAT AND I HAD 2582 01:59:08,425 --> 01:59:10,160 A WONDERFUL YEAR AND A HALF, TWO 2583 01:59:10,160 --> 01:59:11,962 YEARS WORKING IN THE INFECTION 2584 01:59:11,962 --> 01:59:14,031 PARASITE LAB AND REALLY ENJOYING 2585 01:59:14,031 --> 01:59:16,266 MANY, MANY FRIEND, OLD FRIEND 2586 01:59:16,266 --> 01:59:20,103 AND NEW FRIEND I MET THERE. 2587 01:59:20,103 --> 01:59:21,939 THAT I ALSO KNEW FOR A LONG TIME 2588 01:59:21,939 --> 01:59:23,440 ENTICED ME TO MOVE TO FREDERICK 2589 01:59:23,440 --> 01:59:31,848 AND THEN EVENTUALLY TO BETHESDA, 2590 01:59:31,848 --> 01:59:33,817 AND -- FOR ALMOST 20 YEARS. 2591 01:59:33,817 --> 01:59:36,687 SO WHAT I WAS DESCRIBING TO YOU 2592 01:59:36,687 --> 01:59:37,821 WORK BETWEEN INFECTION DISEASE 2593 01:59:37,821 --> 01:59:39,122 AND CANCER REALLY START MAKE ME 2594 01:59:39,122 --> 01:59:40,757 AND SOME OF MY COLLABORATORS TO 2595 01:59:40,757 --> 01:59:43,660 REALLY PUSH THIS IDEA, AND 2596 01:59:43,660 --> 01:59:45,762 REALLY THINKING ABOUT LEARNING 2597 01:59:45,762 --> 01:59:50,033 FROM THE TWO SYSTEM HOW TO -- 2598 01:59:50,033 --> 01:59:51,902 INFECTION DISEASE -- UNDERSTAND 2599 01:59:51,902 --> 01:59:56,707 CANCER IMMUNITY, THE FACT THAT 2600 01:59:56,707 --> 01:59:58,775 ANY INFECTION, ACUTE INFECTION 2601 01:59:58,775 --> 02:00:02,012 IS ACTIVE TO ELIMINATE THE 2602 02:00:02,012 --> 02:00:02,879 PATHOGEN, BUT THEN YOU SHOULD 2603 02:00:02,879 --> 02:00:06,483 MOVE THE INFLAMMATION RESOLUTION 2604 02:00:06,483 --> 02:00:10,587 INFECTION INFLAMMATION IMMUNITY 2605 02:00:10,587 --> 02:00:12,356 TO GO BACK TO TISSUE 2606 02:00:12,356 --> 02:00:13,223 HOMEOSTASIS. 2607 02:00:13,223 --> 02:00:13,957 IN TUMOR IT'S ACTUALLY THE 2608 02:00:13,957 --> 02:00:14,725 OPPOSITE. 2609 02:00:14,725 --> 02:00:17,961 THIS MECHANISM OR RESOLUTION OF 2610 02:00:17,961 --> 02:00:20,163 INFLAMMATION ARE THE ONE THAT 2611 02:00:20,163 --> 02:00:24,768 INDUCE CHRONIC INFLAMMATION, 2612 02:00:24,768 --> 02:00:25,636 INDUCE -- AND BLOCK IMMUNE 2613 02:00:25,636 --> 02:00:27,337 AGAINST THE TUMOR, AND IN TUMOR, 2614 02:00:27,337 --> 02:00:29,406 WE WANT TO DO THE OPPOSITE, WE 2615 02:00:29,406 --> 02:00:33,377 ACTUALLY WANT TO REACTIVATE THE 2616 02:00:33,377 --> 02:00:34,811 IMMUNE SYSTEM AND INFLAMMATION 2617 02:00:34,811 --> 02:00:36,446 IN A WAY TO KILL THE TUMOR 2618 02:00:36,446 --> 02:00:38,849 CELLS. 2619 02:00:38,849 --> 02:00:41,418 AND THE LAST 10 OR 12 YEARS WHAT 2620 02:00:41,418 --> 02:00:44,154 REALLY MY LAB REALLY STARTED 2621 02:00:44,154 --> 02:00:48,859 FOCUSING WAS A NEW ASPECT IN 2622 02:00:48,859 --> 02:00:49,993 MICROBIOLOGY INFECTIOUS DISEASES 2623 02:00:49,993 --> 02:00:53,930 REALLY WAS THE ROLE OF THE 2624 02:00:53,930 --> 02:00:55,999 COMMENSAL -- IS MICROBIOME. 2625 02:00:55,999 --> 02:00:58,535 AND REALLY WE'VE BEEN INTERESTED 2626 02:00:58,535 --> 02:01:00,537 TO START STUDYING CANCER AS A 2627 02:01:00,537 --> 02:01:02,172 DISEASE OF THE METAORGANISM, SO 2628 02:01:02,172 --> 02:01:08,345 THE ORGANISM LIKE OTHER ANIMAL 2629 02:01:08,345 --> 02:01:11,181 DPSH BY THEIR OWN CELLS TOGETHER 2630 02:01:11,181 --> 02:01:13,050 WITH THE COMMAND CELL MICROBE. 2631 02:01:13,050 --> 02:01:14,685 WE KNOW THIS MICROBIOME IS 2632 02:01:14,685 --> 02:01:15,619 REALLY IMPORTANT FOR THE 2633 02:01:15,619 --> 02:01:20,223 SURVIVAL OF THE METAORGANNISM. 2634 02:01:20,223 --> 02:01:25,162 THE CROSSTALK BETWEEN THE 2635 02:01:25,162 --> 02:01:26,830 MICROBIOTA REGULATE MANY 2636 02:01:26,830 --> 02:01:30,133 PHYSICAL FUNCTION, THIS INCLUDES 2637 02:01:30,133 --> 02:01:32,869 IMMUNITY, MECHANICAL FUNCTION 2638 02:01:32,869 --> 02:01:33,203 METABOLISM. 2639 02:01:33,203 --> 02:01:34,771 SO WE REALLY WANTED TO STUDY 2640 02:01:34,771 --> 02:01:36,306 THIS IN CANCER, AND WE WORK WITH 2641 02:01:36,306 --> 02:01:37,841 MANY OF THE INVESTIGATOR OF NIH, 2642 02:01:37,841 --> 02:01:40,510 I WOULD SAY THAT PARTICULAR 2643 02:01:40,510 --> 02:01:43,914 WHICH WITH JULIE SEGRA AND -- IN 2644 02:01:43,914 --> 02:01:48,985 REALLY TRYING TO PUSH THE STURDY 2645 02:01:48,985 --> 02:01:51,388 OF THE MICROBIOME AS NEW 2646 02:01:51,388 --> 02:01:52,155 FRONTIER OF RESEARCH. 2647 02:01:52,155 --> 02:01:55,392 I THINK THIS TRANS NIEH ATTEMPT 2648 02:01:55,392 --> 02:01:57,594 DIDN'T WORK VERY WELL BUT EASILY 2649 02:01:57,594 --> 02:02:00,764 THE CONTRIBUTION OF JULIE SAGER 2650 02:02:00,764 --> 02:02:04,101 TOGETHER WITH -- THE FIELD AS 2651 02:02:04,101 --> 02:02:06,503 REALLY BEING GREAT FUNDAMENTALLY 2652 02:02:06,503 --> 02:02:10,774 AS MAY EVENTUALLY BECOME -- OUR 2653 02:02:10,774 --> 02:02:14,277 WORK TOGETHER WITH -- IN 2654 02:02:14,277 --> 02:02:15,645 COLLABORATION WITH ME ON OUR OWN 2655 02:02:15,645 --> 02:02:17,814 RECEIVED SOME INTEREST AND I 2656 02:02:17,814 --> 02:02:20,517 ACTUALLY WAS QUITE EXCITED WHEN 2657 02:02:20,517 --> 02:02:21,485 NATIONAL GEOGRAPHIC REPORTED OUR 2658 02:02:21,485 --> 02:02:21,685 WORK. 2659 02:02:21,685 --> 02:02:24,087 I NEVER THOUGHT NATIONAL 2660 02:02:24,087 --> 02:02:24,588 GEOGRAPHIC WOULD BECOME 2661 02:02:24,588 --> 02:02:28,058 INTERESTED IN OUR WORK. 2662 02:02:28,058 --> 02:02:34,364 WHAT WAS NOT SO NICE, THAT THE 2663 02:02:34,364 --> 02:02:36,633 SECTION -- WAS NOT EXACTLY 2664 02:02:36,633 --> 02:02:37,300 ROCKET SCIENCE. 2665 02:02:37,300 --> 02:02:38,602 THEN I REMEMBERED THAT MY SON 2666 02:02:38,602 --> 02:02:40,404 GAVE ME A PRESENT A FEW YEARS 2667 02:02:40,404 --> 02:02:42,806 BEFORE AND I THINK HE WAS VERY 2668 02:02:42,806 --> 02:02:44,241 PROPHETIC AND HE GAVE ME THIS 2669 02:02:44,241 --> 02:02:45,609 COFFEE CUP, AND I STILL HAVE IT 2670 02:02:45,609 --> 02:02:46,576 ON MY DESK. 2671 02:02:46,576 --> 02:02:49,212 I THINK IT'S A GOOD REMINDER FOR 2672 02:02:49,212 --> 02:02:55,218 HOW WE SHOULD LOOK AT SCIENCE. 2673 02:02:55,218 --> 02:02:57,354 OKAY, SO MICROBIOME IS NOW SEEN 2674 02:02:57,354 --> 02:03:02,559 AS A MAJOR PLAYER IN CANCER, 2675 02:03:02,559 --> 02:03:03,193 IT'S -- INSTABILITY, DEFINITELY 2676 02:03:03,193 --> 02:03:08,165 IN FAVORING -- TUMOR GROW, BUT 2677 02:03:08,165 --> 02:03:10,834 ALSO AS A MAY YOUR ROLE IN 2678 02:03:10,834 --> 02:03:15,005 REGULATION OF CANCER THERAPY. 2679 02:03:15,005 --> 02:03:16,206 THIS HAS BEEN GOING FOR QUITE A 2680 02:03:16,206 --> 02:03:18,942 LONG TIME, IT WAS ONLY 2681 02:03:18,942 --> 02:03:20,777 '22 THAT -- FINALLY PUT THE 2682 02:03:20,777 --> 02:03:26,550 MICROBIOME AS ONE OF THE 2683 02:03:26,550 --> 02:03:35,659 CANCER -- SO WHAT WE DID WAS THE 2684 02:03:35,659 --> 02:03:37,360 WANT TO KNOW IF THE CANCER 2685 02:03:37,360 --> 02:03:39,396 THERAPY COULD BE AFFECTED BY THE 2686 02:03:39,396 --> 02:03:39,729 MICROBIOME. 2687 02:03:39,729 --> 02:03:44,267 AND WE ESTABLISHED THE GUT 2688 02:03:44,267 --> 02:03:47,003 MYOBIOME -- CANCER THERAPY 2689 02:03:47,003 --> 02:03:48,772 MOSTLY BY MODULATE ANTITUMOR 2690 02:03:48,772 --> 02:03:51,308 RESPONSE, MONITORING -- IMMUNITY 2691 02:03:51,308 --> 02:03:54,044 BUT DIRECTLY INDIRECTLY -- 2692 02:03:54,044 --> 02:03:54,578 IMMUNITY. 2693 02:03:54,578 --> 02:03:57,447 AND OUR PAPER IN WHICH THE MAJOR 2694 02:03:57,447 --> 02:03:58,615 PLAYER -- NOT SHOWN IN THIS 2695 02:03:58,615 --> 02:04:00,884 PICTURE, HE WAS ALREADY BACK TO 2696 02:04:00,884 --> 02:04:04,821 JAPAN, BUT -- AND HIS -- IN 2013 2697 02:04:04,821 --> 02:04:10,360 WAS TO SHOW THAT IF YOU WANT 2698 02:04:10,360 --> 02:04:13,530 TO -- OR IMMUNOTHERAPY USING -- 2699 02:04:13,530 --> 02:04:18,568 YOU NEED THE PRESENCE OF THE GUT 2700 02:04:18,568 --> 02:04:18,869 MICROBIOTA. 2701 02:04:18,869 --> 02:04:20,203 AND WITHIN GUT MICROBIOTA, YOU 2702 02:04:20,203 --> 02:04:25,141 NEED CERTAIN SPECIES THAT WERE 2703 02:04:25,141 --> 02:04:27,744 FAVORTIVE RESPONSE. 2704 02:04:27,744 --> 02:04:30,881 -- FEW YEARS BEFORE LIKELY A 2705 02:04:30,881 --> 02:04:34,484 DIFFERENT SITUATION -- SHOW THAT 2706 02:04:34,484 --> 02:04:37,787 LPS AND MICROBIOME -- SOME ROLE 2707 02:04:37,787 --> 02:04:41,558 BUT AT THE SAME TIME, WE PUBLISH 2708 02:04:41,558 --> 02:04:47,697 IN 2013LAURENCE ZI TV OGEL, LONG 2709 02:04:47,697 --> 02:04:49,065 TIME FRIEND OF MINE, IN 2710 02:04:49,065 --> 02:04:50,133 PITTSBURGH, PUBLISHED IN PARIS 2711 02:04:50,133 --> 02:04:54,671 THE ROLE OF THE MICROBIOME AND 2712 02:04:54,671 --> 02:04:57,307 OTHER CHEMOTHERAPY IN 2713 02:04:57,307 --> 02:04:57,974 CYCLOPHOSPHAMIDE. 2714 02:04:57,974 --> 02:05:00,377 THE MAJOR INTEREST IN THE 2715 02:05:00,377 --> 02:05:05,348 COMMUNITY REALLY COME WHEN 2716 02:05:05,348 --> 02:05:06,449 LAURENCE AS WELL AS -- SHOW IN 2717 02:05:06,449 --> 02:05:11,254 MICE THAT THE MICROBIOTA PLAY A 2718 02:05:11,254 --> 02:05:16,259 ROLE NOT ONLY IN CHEMOTHERAPY 2719 02:05:16,259 --> 02:05:17,994 BUT ALSO IN IMMUNOTHERAPY WITH 2720 02:05:17,994 --> 02:05:23,967 IMMUNE CHECKPOINT INHIBITORS IN 2721 02:05:23,967 --> 02:05:26,269 CTLA4 OR ANTIPD-1. 2722 02:05:26,269 --> 02:05:32,509 OUR PAPER AND LAURECE'S PAPER IS 2723 02:05:32,509 --> 02:05:35,946 ALSO LISTED AS IN SCIENCE 2013, 2724 02:05:35,946 --> 02:05:37,914 SO WE CONTINUE WORK IN THE LAST 2725 02:05:37,914 --> 02:05:39,783 10 YEARS IN MICE IN CLINICAL 2726 02:05:39,783 --> 02:05:43,219 WORK, AND WE STUDY TOGETHER WITH 2727 02:05:43,219 --> 02:05:45,956 MANY OTHER INVESTIGATORS SOME OF 2728 02:05:45,956 --> 02:05:51,227 THE M -- IN WHICH SOME BACTERIAN 2729 02:05:51,227 --> 02:05:54,331 THE LEFT MAY FAVOR RESPONSE TO 2730 02:05:54,331 --> 02:05:55,765 ANTI-PD-1 BUT -- WOULD PLAY A 2731 02:05:55,765 --> 02:05:58,335 MAJOR ROLE IS OUR OTHER TYPE OF 2732 02:05:58,335 --> 02:06:00,070 BACTERIA, MOSTLY GRAM-NEGATIVE, 2733 02:06:00,070 --> 02:06:03,273 SEEMS TO HAVE AN INHIBITORY 2734 02:06:03,273 --> 02:06:05,075 EFFECT ON THE RESPONSE OF PD-1 2735 02:06:05,075 --> 02:06:07,143 AND THE QUESTION IS WHAT DO WE 2736 02:06:07,143 --> 02:06:08,878 WANT TO TARGET FOR THERAPY, THE 2737 02:06:08,878 --> 02:06:11,047 POSITIVE EFFECT OR THE 2738 02:06:11,047 --> 02:06:13,450 ELIMINATING NEGATIVE EFFECT. 2739 02:06:13,450 --> 02:06:19,222 NOW, IF THE MICROBIOME EFFECT -- 2740 02:06:19,222 --> 02:06:20,490 THE RESPONSE TO CANCER THERAPY 2741 02:06:20,490 --> 02:06:24,227 AND BECAUSE WE CAN CHARACTERIZE 2742 02:06:24,227 --> 02:06:25,128 THE MICROBIOME IN THE IF YOU 2743 02:06:25,128 --> 02:06:31,735 TELL OGUT OF THEPATIENT, CAN WEE 2744 02:06:31,735 --> 02:06:33,436 PATIENT WILL RESPOND BASED ON 2745 02:06:33,436 --> 02:06:34,304 THE BABBLING TIER YA IN THE GUT 2746 02:06:34,304 --> 02:06:38,908 BACTERIA IN THE GUT OF THE 2747 02:06:38,908 --> 02:06:39,342 PATIENT. 2748 02:06:39,342 --> 02:06:42,312 EACH INDIVIDUAL IS ABOUT 2749 02:06:42,312 --> 02:06:43,179 COUPLE -- MORE DIFFERENT BACK 2750 02:06:43,179 --> 02:06:44,447 FEAR YA SPECIES IN THE GUT AND 2751 02:06:44,447 --> 02:06:48,151 THERE ARE SEVERAL THOUSAND -- 2752 02:06:48,151 --> 02:06:49,252 DISTRIBUTE IN DIFFERENT BACTERIA 2753 02:06:49,252 --> 02:06:50,887 AND EACH PERSON IS REALLY 2754 02:06:50,887 --> 02:06:53,056 DIFFERENT AND ITS GEOGRAPHIC 2755 02:06:53,056 --> 02:06:54,591 LOCALITY IS A DIFFERENT 2756 02:06:54,591 --> 02:06:55,258 MICROBIOME. 2757 02:06:55,258 --> 02:06:56,760 SO WHEN YOU TRY TO USE A MACHINE 2758 02:06:56,760 --> 02:06:58,695 LEARNING TO PREDICT A RESPONSE, 2759 02:06:58,695 --> 02:06:59,796 TRAIN THE MACHINE LEARNING WITH 2760 02:06:59,796 --> 02:07:04,267 THE DATA FROM ONE -- AND PREDICT 2761 02:07:04,267 --> 02:07:07,937 THE RESPONSE IN ANOTHER ONE IS 2762 02:07:07,937 --> 02:07:08,705 COMPLETELY FAIL. 2763 02:07:08,705 --> 02:07:12,308 BUT IF YOU DO TRAIN YOUR MODEL 2764 02:07:12,308 --> 02:07:15,612 USING ALL THE DIFFERENT CORE 2765 02:07:15,612 --> 02:07:17,814 TOGETHER LEAVING OUT ONLY ONE 2766 02:07:17,814 --> 02:07:18,848 CORE, THEN THINGS START TO BE 2767 02:07:18,848 --> 02:07:20,717 BETTER AND WE CAN GET A 2768 02:07:20,717 --> 02:07:22,018 REASONABLE HIGH ACCURACY IN 2769 02:07:22,018 --> 02:07:23,153 PREDICTION OF THE RESPONSE. 2770 02:07:23,153 --> 02:07:26,556 THERE ARE ALWAYS A LOT OF -- IN 2771 02:07:26,556 --> 02:07:28,091 THIS WORK DESPITE THE DIFFERENT 2772 02:07:28,091 --> 02:07:29,659 QUALITY OF MELANOMA PATIENT AND 2773 02:07:29,659 --> 02:07:30,760 THERE'S REALLY A LOT OF WORK IN 2774 02:07:30,760 --> 02:07:32,629 OUR LAB AND MANY OTHER LABS TO 2775 02:07:32,629 --> 02:07:36,833 TRY TO DEFINE MIC MICROBIOME 2776 02:07:36,833 --> 02:07:37,500 BIOMARKER FOR PREDICTION OF THE 2777 02:07:37,500 --> 02:07:39,135 RESPONSE. 2778 02:07:39,135 --> 02:07:43,139 BUT THE OTHER THINGS IS THE -- 2779 02:07:43,139 --> 02:07:45,108 CAN WE USE -- IN THERAPY, CAN WE 2780 02:07:45,108 --> 02:07:47,544 TARGET THE MICROBIOME TO IMPROVE 2781 02:07:47,544 --> 02:07:49,946 THERAPY RESPONSE. 2782 02:07:49,946 --> 02:07:51,781 AND WHAT WE DID AND ALL OUR 2783 02:07:51,781 --> 02:07:54,084 CLINICAL WORK IS REALLY BEEN A 2784 02:07:54,084 --> 02:07:54,951 WONDERFUL -- MANY A 2785 02:07:54,951 --> 02:07:57,187 COLLABORATION NOW WITH SHOWING 2786 02:07:57,187 --> 02:08:06,062 HERE THE WORK OF DEVAR AND -- IN 2787 02:08:06,062 --> 02:08:07,030 PITTSBURGH, AND WHAT WE WERE 2788 02:08:07,030 --> 02:08:12,335 ABLE TO SHOW THAT IF WE GIVE A 2789 02:08:12,335 --> 02:08:14,537 FECAL MICROBIOTA TRANSPLANT FROM 2790 02:08:14,537 --> 02:08:17,173 A PATIENT RESPOND TO ANTIPD-1 TO 2791 02:08:17,173 --> 02:08:18,241 PATIENT THAT HAVE COMPLETELY 2792 02:08:18,241 --> 02:08:21,478 FAILED TO RESPOND TO ANTIPD-1, 2793 02:08:21,478 --> 02:08:25,615 WE GET A REVERSION OVERCOME THIS 2794 02:08:25,615 --> 02:08:27,684 PD-1 RESISTANCE IN APPROXIMATELY 2795 02:08:27,684 --> 02:08:28,885 40% OF THE PATIENTS. 2796 02:08:28,885 --> 02:08:31,755 THERE ARE NOW THREE OR FOUR 2797 02:08:31,755 --> 02:08:34,023 STUDY IN OTHER GROUP THAT'S 2798 02:08:34,023 --> 02:08:36,760 SHOWN ABOUT THE SAME RESULT THAT 2799 02:08:36,760 --> 02:08:41,664 IS VERY PROMISING. 2800 02:08:41,664 --> 02:08:42,699 BUT WHAT ACTUALLY WAS HAPPENING 2801 02:08:42,699 --> 02:08:44,901 JUST A FEW DAYS AGO, A 2802 02:08:44,901 --> 02:08:46,002 PRELIMINARY STUDY WAS REPORTED 2803 02:08:46,002 --> 02:08:49,272 BY GROUP IN ITALY THAT DID FIRST 2804 02:08:49,272 --> 02:08:51,007 RANDOMIZED TRIAL, IT IS ACTUALLY 2805 02:08:51,007 --> 02:08:55,278 QUITE SIGNIFICANT EFFECT OF THE 2806 02:08:55,278 --> 02:08:56,780 FECAL MICROBIOTA TRANSPLANT IN 2807 02:08:56,780 --> 02:08:57,881 IMPROVING THE RESPONSE IN 2808 02:08:57,881 --> 02:08:58,982 PATIENTS THAT FAILED THERAPY 2809 02:08:58,982 --> 02:08:59,949 BEFORE THE TRANSPLANT. 2810 02:08:59,949 --> 02:09:01,785 AND THE OTHER THING THAT WE WORK 2811 02:09:01,785 --> 02:09:04,654 AND THAT WE ARE TRYING TO FIGURE 2812 02:09:04,654 --> 02:09:06,790 OUT, ARE THERE WAY TO CHANGE THE 2813 02:09:06,790 --> 02:09:09,392 MICROBIOTA, AND ONE WAY THAT IS 2814 02:09:09,392 --> 02:09:12,662 POSSIBLE TO DO THAT IS WITH 2815 02:09:12,662 --> 02:09:17,133 DIET, AND TOGETHER WORKING WITH 2816 02:09:17,133 --> 02:09:19,869 JENNIFER WARGO, WE'VE BEEN ABLE 2817 02:09:19,869 --> 02:09:23,006 TO SHOW THAT PATIENT WITH HIGH 2818 02:09:23,006 --> 02:09:24,207 FIBER DIET ARE MUCH MORE 2819 02:09:24,207 --> 02:09:26,943 RESPONSE SIEVE TO ANTIPD-1 2820 02:09:26,943 --> 02:09:29,112 THERAPY, WE COMPLETELY 2821 02:09:29,112 --> 02:09:29,979 REPRODUCED THOSE RESULTS IN MICE 2822 02:09:29,979 --> 02:09:32,816 AND SHOWED THAT THE FIBER OF THE 2823 02:09:32,816 --> 02:09:34,784 DIET IS AFFECTING PD-1 RESPONSE 2824 02:09:34,784 --> 02:09:39,556 BY CHANGING THE MICROBIOTA 2825 02:09:39,556 --> 02:09:41,858 COMPOSITION IN THE -- AT LEAST 2826 02:09:41,858 --> 02:09:48,097 IN THE MOUSE ENVIRONMENT. 2827 02:09:48,097 --> 02:09:50,333 SO I'VE BEEN DISCUSSING LIKE MY 2828 02:09:50,333 --> 02:09:52,101 TWO COLLEAGUES BEFORE ME MY 2829 02:09:52,101 --> 02:09:52,969 YEARS IN SCIENCE. 2830 02:09:52,969 --> 02:09:56,039 I WOULD LIKE TO LIST EACH 2831 02:09:56,039 --> 02:09:59,042 PERSON, ITS CONTRIBUTION, I 2832 02:09:59,042 --> 02:10:00,243 DON'T HAVE A PICTURE OF EVERYONE 2833 02:10:00,243 --> 02:10:01,444 I'VE WORKED WITH, I TRY TO 2834 02:10:01,444 --> 02:10:02,879 COLLECT AS MANY AS POSSIBLE, 2835 02:10:02,879 --> 02:10:04,047 SOME OF YOU MAY KNOW SOME OF 2836 02:10:04,047 --> 02:10:07,650 THEM, ESPECIALLY THE NIH PEOPLE 2837 02:10:07,650 --> 02:10:11,888 BUT IT'S BEEN A LONG JOURNEY, A 2838 02:10:11,888 --> 02:10:13,189 REALLY REWARDING ONE AND I'M 2839 02:10:13,189 --> 02:10:15,225 THANKFUL TO THE NIH FOR THE LAST 2840 02:10:15,225 --> 02:10:16,092 20 YEARS AND BE ABLE TO DO THIS 2841 02:10:16,092 --> 02:10:16,526 TYPE OF WORK. 2842 02:10:16,526 --> 02:10:17,093 THANK YOU VERY MUCH. 2843 02:10:17,093 --> 02:10:22,899 [APPLAUSE] 2844 02:10:22,899 --> 02:10:23,666 >> THANK YOU. 2845 02:10:23,666 --> 02:10:24,601 KNOW THE ROUTINE. 2846 02:10:24,601 --> 02:10:25,902 IF YOU HAVE QUESTIONS, AT THE 2847 02:10:25,902 --> 02:10:32,575 MICS. 2848 02:10:32,575 --> 02:10:36,079 >> YOU KNOW, IT INTERESTING TO 2849 02:10:36,079 --> 02:10:40,516 ME THAT YOUR JOURNEY HAS TAKEN 2850 02:10:40,516 --> 02:10:45,588 YOU FROM THE ENDOGENOUS IMMUNE 2851 02:10:45,588 --> 02:10:46,756 SYSTEM AND THE CELLS THAT 2852 02:10:46,756 --> 02:10:49,225 MEDIATE IT, AND I THINK WE ALL 2853 02:10:49,225 --> 02:10:55,598 THINK OF AS BEING THERE TO FIGHT 2854 02:10:55,598 --> 02:11:00,403 THIS FOE, THE EXOGENOUS 2855 02:11:00,403 --> 02:11:03,940 MICROBIAL UNIVERSE, ALL THE WAY 2856 02:11:03,940 --> 02:11:05,141 ALMOST FULL CIRCLE TO THE POINT 2857 02:11:05,141 --> 02:11:08,011 WHERE YOU'RE STUDYING THE 2858 02:11:08,011 --> 02:11:10,914 PEACEFUL COEXISTENCE AND MAYBE 2859 02:11:10,914 --> 02:11:13,316 COLLABORATION WITH THE 2860 02:11:13,316 --> 02:11:16,486 MICROBIOME. 2861 02:11:16,486 --> 02:11:19,222 IS THERE A DANGER OF THE 2862 02:11:19,222 --> 02:11:22,892 ENDOGENOUS IMMUNE SYSTEM KILLING 2863 02:11:22,892 --> 02:11:25,528 OFF SOME OF THESE -- I MEAN, ARE 2864 02:11:25,528 --> 02:11:30,133 THERE DISORDERS THAT RESULT FROM 2865 02:11:30,133 --> 02:11:33,870 SEEING THE MICROBIOME AS FOE, OR 2866 02:11:33,870 --> 02:11:37,507 IS THERE SOMETHING THAT PROTECTS 2867 02:11:37,507 --> 02:11:40,610 THESE PARTICULAR ORGANISMS FROM 2868 02:11:40,610 --> 02:11:43,313 ATTACK BY THE HUMAN IMMUNE 2869 02:11:43,313 --> 02:11:43,546 SYSTEM? 2870 02:11:43,546 --> 02:11:49,185 >> OBVIOUSLY, I MEAN, WE EVOLVE 2871 02:11:49,185 --> 02:11:50,820 AS AN ECOLOGICAL SYSTEM AND WE 2872 02:11:50,820 --> 02:11:52,555 HAVE MOST OF THE TIME PRETTY 2873 02:11:52,555 --> 02:11:54,991 GOOD. 2874 02:11:54,991 --> 02:11:58,027 THERE ARE INNATE AND IMMUNE 2875 02:11:58,027 --> 02:12:00,563 MECHANISMS THAT REGULATE THAT. 2876 02:12:00,563 --> 02:12:02,265 THE EPITHELIAL BARRIER ALWAYS 2877 02:12:02,265 --> 02:12:03,800 PLAYS A MAJOR PHYSICAL ROLE IN 2878 02:12:03,800 --> 02:12:06,002 AVOIDING THAT. 2879 02:12:06,002 --> 02:12:10,940 BUT SOMETIMES THINGS GO WRONG 2880 02:12:10,940 --> 02:12:12,308 AND YOU HAVE TWO TYPE OF 2881 02:12:12,308 --> 02:12:12,742 PATHOLOGY. 2882 02:12:12,742 --> 02:12:16,079 ONE IS WHAT THEY CALL -- THEY 2883 02:12:16,079 --> 02:12:17,447 NORMALLY ARE OKAY, BUT IN 2884 02:12:17,447 --> 02:12:20,516 CERTAIN SITUATIONS, THEY MIGHT 2885 02:12:20,516 --> 02:12:23,319 BECOME A PATHOGEN. 2886 02:12:23,319 --> 02:12:26,923 -- TYPICAL ONE, WE ALL -- 2887 02:12:26,923 --> 02:12:29,759 DIFFICILE, BUT IF IT GROW IS NOT 2888 02:12:29,759 --> 02:12:30,960 CONTROLLED IT BECOMES A MAJOR 2889 02:12:30,960 --> 02:12:31,227 PATHOGEN. 2890 02:12:31,227 --> 02:12:32,261 THE OTHER ONE IS THE OPPOSITE 2891 02:12:32,261 --> 02:12:35,765 WHEN OUR SYSTEM IS BECOMING TOO 2892 02:12:35,765 --> 02:12:37,033 ACTIVATED AGAINST THE COMMENSAL 2893 02:12:37,033 --> 02:12:40,103 AND THEN OBVIOUSLY THE MAJOR 2894 02:12:40,103 --> 02:12:41,104 DISEASE YOU CAN THINK IN THIS 2895 02:12:41,104 --> 02:12:43,573 CASE IS INFLAMMATORY BOWEL 2896 02:12:43,573 --> 02:12:45,842 DISEASE, CROHN'S DISEASE, AND 2897 02:12:45,842 --> 02:12:46,843 ULCERATIVE COLITIS. 2898 02:12:46,843 --> 02:12:48,277 >> WONDERFUL. 2899 02:12:48,277 --> 02:12:49,579 WONDERFUL. 2900 02:12:49,579 --> 02:12:55,351 YOU KNOW, I THINK ALL OF THE 2901 02:12:55,351 --> 02:12:56,452 TALKS WE'VE HEARD TODAY REALLY 2902 02:12:56,452 --> 02:13:00,923 GIVE US A SENSE THAT THERE'S 2903 02:13:00,923 --> 02:13:02,125 ALWAYS A NEXT QUESTION. 2904 02:13:02,125 --> 02:13:07,764 I MEAN, I THINK ESPECIALLY THE 2905 02:13:07,764 --> 02:13:13,436 WORK WE'VE JUST HEARD, YOU KNOW, 2906 02:13:13,436 --> 02:13:16,305 THERE IS SOMETHING ABOUT SCIENCE 2907 02:13:16,305 --> 02:13:20,109 THAT KEEPS ONE INTELLECTUALLY 2908 02:13:20,109 --> 02:13:23,813 CURIOUS FOR A LIFETIME, AND THAT 2909 02:13:23,813 --> 02:13:26,215 ALWAYS MAKES THERE BE A NEXT 2910 02:13:26,215 --> 02:13:26,883 QUESTION. 2911 02:13:26,883 --> 02:13:28,618 SO THANK YOU SO MUCH. 2912 02:13:28,618 --> 02:13:29,218 >> THANK YOU. 2913 02:13:29,218 --> 02:13:34,757 [APPLAUSE] 2914 02:13:34,757 --> 02:13:38,795 >> WELL, BEFORE WE GO OUT FOR 2915 02:13:38,795 --> 02:13:40,863 OUR RECEPTION, I HESITATE TO SAY 2916 02:13:40,863 --> 02:13:42,265 THIS IS A TRADITION, THIS IS 2917 02:13:42,265 --> 02:13:45,535 ONLY THE SECOND TIME I'VE DONE 2918 02:13:45,535 --> 02:13:47,737 THIS, BUT LAST YEAR, CHRIS 2919 02:13:47,737 --> 02:13:49,172 ACTUALLY COMMENTED THAT I DID A 2920 02:13:49,172 --> 02:13:52,208 SORT OF A SUPPLETIVE TOUR, THREE 2921 02:13:52,208 --> 02:13:54,010 SENTENCES, THAT TIED TOGETHER 2922 02:13:54,010 --> 02:13:55,812 THREE TALKS THAT SEEMED AT FACE 2923 02:13:55,812 --> 02:13:57,980 VALUE TO HAVE ABSOLUTELY NOTHING 2924 02:13:57,980 --> 02:14:03,653 TO DO WITH ONE ANOTHER. 2925 02:14:03,653 --> 02:14:05,521 AND THIS YEAR, WHAT STRUCK ME 2926 02:14:05,521 --> 02:14:10,827 WAS THE SORT OF CAREER TAKE-HOME 2927 02:14:10,827 --> 02:14:12,595 LESSONS THAT PERVADED REALLY ALL 2928 02:14:12,595 --> 02:14:15,865 THREE OF THE TALKS THAT WE'VE 2929 02:14:15,865 --> 02:14:19,435 HEARD SO I'VE COME UP WITH FIVE 2930 02:14:19,435 --> 02:14:21,104 SORT OF, I DON'T KNOW, LIFE AND 2931 02:14:21,104 --> 02:14:22,538 CAREER LESSONS THAT I WANT TO 2932 02:14:22,538 --> 02:14:25,441 SHARE WITH YOU, AT LEAST ONE OF 2933 02:14:25,441 --> 02:14:27,310 WHICH MAY GET ME IN A LOT OF 2934 02:14:27,310 --> 02:14:31,781 TROUBLE. 2935 02:14:31,781 --> 02:14:34,417 THE FIRST IS, ONE SHOULDN'T BE 2936 02:14:34,417 --> 02:14:36,986 AFRAID TO STICK ONE'S NOSE WHERE 2937 02:14:36,986 --> 02:14:38,955 SOME WOULD SAY IT DEFINITELY 2938 02:14:38,955 --> 02:14:40,723 DOESN'T BELONG. 2939 02:14:40,723 --> 02:14:43,893 I THINK THAT IS SOME OF THE 2940 02:14:43,893 --> 02:14:46,295 EXCITEMENT OF SCIENCE. 2941 02:14:46,295 --> 02:14:48,698 THE SECOND IS THAT ONE SHOULD 2942 02:14:48,698 --> 02:14:53,169 HASTEN TO JUXTAPOSE FIELDS THAT 2943 02:14:53,169 --> 02:14:54,370 NO SANE PERSON WOULD EVER PUT 2944 02:14:54,370 --> 02:15:00,676 TOGETHER. 2945 02:15:00,676 --> 02:15:01,777 THE THIRD IS THAT YOU SHOULD 2946 02:15:01,777 --> 02:15:03,212 ALWAYS CHECK YOUR WORK AND 2947 02:15:03,212 --> 02:15:04,280 PARTICULARLY YOUR SPELLING 2948 02:15:04,280 --> 02:15:08,351 BEFORE YOU HAND YOUR WORK IN. 2949 02:15:08,351 --> 02:15:09,752 THE FOURTH, AND THIS MAY BE THE 2950 02:15:09,752 --> 02:15:12,822 PROBLEMATIC ONE, IS THAT FOREIGN 2951 02:15:12,822 --> 02:15:15,892 POLITICAL INTERFERENCE EXTENDS 2952 02:15:15,892 --> 02:15:17,727 TO INTERLEUKIN NAMING 2953 02:15:17,727 --> 02:15:22,331 CONVENTIONS. 2954 02:15:22,331 --> 02:15:23,533 AND THE FIFTH, AND THIS IS THE 2955 02:15:23,533 --> 02:15:25,468 CRUX OF THE WHOLE THING, AND 2956 02:15:25,468 --> 02:15:26,402 AGAIN, PERVADED ALL THREE OF THE 2957 02:15:26,402 --> 02:15:29,839 TALKS TODAY, YOU SHOULD USE YOUR 2958 02:15:29,839 --> 02:15:32,008 SCIENCE TO FORM A BROAD AND DEEP 2959 02:15:32,008 --> 02:15:35,478 NETWORK OF FRIENDS AND A LEGACY 2960 02:15:35,478 --> 02:15:36,579 OF IMPACT. 2961 02:15:36,579 --> 02:15:40,850 SO PLEASE JOIN US FOR THE 2962 02:15:40,850 --> 02:15:41,951 RECEPTION OUTSIDE, AND JOIN ME 2963 02:15:41,951 --> 02:15:43,319 IN THANKING OUR SPEAKERS ONCE 2964 02:15:43,319 --> 02:15:43,519 AGAIN. 2965 02:15:43,519 --> 02:15:53,696 [APPLAUSE]