>> GOOD AFTERNOON. I HAVE A COUPLE THINGS TO BRING YOUR ATTENTION TO. NEXT WEEK IS THE LAST WEEK OF THE COURSE IN THIS SESSION AND THE TOPIC IS THE ONE THAT WAS SNOWED OUT EARLIER IN THE YEAR. SO YOU'LL SEE ON THE WEBSITE ABOUT THE SPEAKERS AND THE TOPIC WHICH IS A VERY IMPORTANT ONE. IT'S A COMBINATION OF THE ISSUE OF SCREENING FOR MALIGNANT DISEASE BOTH GENOMICALLY AND LOOKING FOR MARKERS AND THE STATE OF PROSTATE CASE PRESENT. THE OTHER SPEAKER HAS ANALYZED THIS FROM THE STAND POINT OF KIND OF ARE WE ARE DOING THE RIGHT THING AND WHERE DO WE DRAW THE THING. IT'S REALLY AN EXTRA PROVOCATIVE ISSUE WHICH IS REALLY AT THE FOREFRONT NOW OF NEW TECHNOLOGIES AND THEIR APPLICATIONS. SO THAT'S NEXT WEEK. THE FINAL EXAM BACK BREAKER OF A COMPUTERIZED EXAM WHICH HAS BEEN PASSED WITH HIGH GRADES BY EVERYONE WHOSE TAKEN IT. SO THOSE OF YOU WHO HAVEN'T AND WANT TO RECEIVE A CERTIFICATE OR PENDING COURSE YOU CAN PICK IT UP ON-LINE ON OUR WEBSITE. OKAY? SO, TODAY'S SUBJECT OF MALARIA AND PARTICULARLY OUR TWO SPEAKERS WHO I WIL BRIEFLY ABOUT IS PARTICULARLY APPROPRIATE FOR THE SORT OF LOGO OF THE BROOKLYN BRIDGE WHICH JUST DISAPPEARED. THOSE TWO GENTLEMEN ARE DISCUSSING, ONE IS SAYING WHEET THE ORIGIN OF MALARIA. WHERE DID SIZE MODE YEAH COME FROM ARE THEY 50,000 YEARS OLD OR A HUNDRED THOUSAND YEARS OLD. AND WHAT'S THE RELATIONSHIP. WHY ARE SOME PEOPLE IN SOME PARTS OF THE WORLD SEEM TO BE RESISTENT. AND OTHERS AREN'T. WHAT CAN WE DO ABOUT IT. IT'S A HUGE PROBLEM BECAUSE EVEN THEN THEY KNEW THAT THIS WAS A GLOBAL PROBLEM. IT'S AT THE END OF THE 19TH CENTURY, AND OF COURSE TODAY WE KNOW THAT THERE ARE SEVERAL HUNDRED MILLION PEOPLE WHO SUFFER FROM MALARIA WITH A FAIRLY HIGH MORTALITY RATE, PARTICULARLY AMONGST YOUNG CHILDRENLY LARGELY IN SUB-SAHARAN AFRICA AND THE TROPICAL AND SUBTROPICAL AREA OF THE WORLD. AND THE OTHER CHAP IS SAYING BUT LOOK THIS THING IS TRANSMIT BY A MOSQUITO. MAYBE WE CAN UNDERSTAND WHAT'S THE INTERACTION BETWEEN THE PARASITE AND THE PRIMARY VECTOR, THE MOSQUITO. IF WE UNDERSTAND SOMETHING ABOUT WHAT HAPPENS IN THE MOSQUITO MAYBE THAT WILL GIVE US SOME CLUES AS TO HOW WE MIGHT EVEN BE ABLE TO TARGET SOME FORMS OF THERAPY. BECAUSE IN THE LAST ANALYSIS, BACK THEN AND EVEN NOW, THERAPY HAS BEEN A TWO-EDGED SWORD. THERE'S AS MUCH PUBLIC HEALTH VENTURES TO DEAL WITH WATER, NETTING OVER BED AT NIGHT BECAUSE OF THE FEMALE ANOMALIES, THE EATING OF HUMAN BLOOD AND BASICALLY THEN PICKS UP THE ONE FORM OF THE PARASITE AND LET IT MULTIPLY AND THEN DELIVERS IT TO THE NEXT VICTIM. THERE ARE DRUGS WHICH ARE EFFECTIVE AND IN SOME OF THE MORE VIRULENT FORMS HAVE BEEN LIFE SAVING. THERE'S HUGE PROBLEMS IN DRUG RESISTANCE AND A NEED TO DEVELOP PERHAPS NEW AGENTS IF YOU KNEW WHAT YOU WERE TARGETING. AND THEN BEHIND ALL OF THIS IS AN INCREDIBLE STORY OF WORLD HISTORY, WARFARE, POVERTY, THE EFFECT THIS DISEASE HAS BEEN PROBABLY MUCH GREATER ON THE HISTORY OF THE WORLD AND MANY OF THE THINGS THAT WE TEMPORARILY THINK ARE SO IMPORTANT. WELL, SO OUR TWO SPEAKERS TODAY CAROLINA BARILLAS-MURY WHO IS HERE AT THE NIH, SHE IS FROM GUATEMALA WHERE SHE GRADUATED IN MEDICINE. AND THEN WENT TO UNIVERSITY OF ARIZONA WHERE SHE RECEIVED A PH.D. DEGREE. AND THEN WAS A POST DOCTORAL FELLOW WITH A VERY FAMOUS PROFESSOR AT HARVARD, OTIS WHO OSCILLATE BETWEEN CAMBRIDGE AND THE EUROPEAN WHERE SHE RECEIVED HER POST DOCTORAL TRAINING. SHE WENT TO THE UNIVERSITY OF COLORADO, COULD BE STATE UNIVERSITY FOR A WHILE AND CAME HERE IN 2003 IN NIAID AND IS NOW A SENIOR INVESTIGATOR IN THE NATIONAL, THE LABORATORY OF MALARIA AND VECTOR RESEARCH AND HEAD OF THE MOSQUITO COMMUNITY AND VECTOR COMPETENCE SECTION. CAROLINA IS HIGHLY HONORED AND MOST RECENTLY, ACTUALLY THERE'S A MISTAKE IN THE SLIDE BECAUSE IT WASN'T YESTERDAY, IT WAS TODAY SHE WAS WELCOMED INTO THE NATIONAL ACADEMY OF SCIENCES. HER STUDIES DEAL WITH THE INTERACTIONS OF THE PLASMADOIUM AND THE CELL OF THE MOSQUITO. HER FOCUS HAS BEEN PRETTY MUCH ON THE MOSQUITO. SO WE ALL CONGRATULATE YOU AND LOOK FORWARD TO YOUR PRESENTATION. [APPLAUSE] OUR SECOND SPEAKER WHICH IS A GUEST VISITING FROM THE UNIVERSITY OF PENNSYLVANIA IS BEATRICE HAHN A PROFESSOR OF MEDICINE IN MICROBIOLOGY. SHE RECEIVED HER MEDICAL DEGREE IN MUNICH AND THEN CAME TO THE NIH H AS A POST DOCTORAL FELLOW IN BOB GALLO'S LABORATORY WHERE SHE LEARNED RETROVIROLOGY WHICH HAS BEEN THE BASIS OF HER RESEARCH. INITIALLY AT THE UNIVERSITY OF ALABAMA, BIRMINGHAM WHERE SHE WAS FOR A RELATIVELY LONG TIME AND THEN MOVED TO THE UNIVERSITY OF PENNSYLVANIA. SHE'S ALSO A MEMBER OF THE NATIONAL ACADEMY. TODAY IS DOUBLE THREAD DAY, WE REALLY HAVE TWO SUPER STARS. HER MAIN INTEREST HAS BEEN IN AT THE ORIGIN, THE EVOLUTIONARY IMPLICATIONS OF THE ORIGIN OF INITIALLY HIV, PARTICULARLY SIMIAN RETROVIRUS AND IN RECENT YEARS TURNED HER ATTENTION TO MALARIA. HER STUDIES WHICH ARE ON THE WEBSITE, I THINK YOU'LL FIND EXTRAORDINARILY EXCITING AND INTERESTING AND I WILL NOT BELABOR THAT ANY FURTHER. SO I THINK BEATRICE YOU'RE GOING TO BE THE FIRST SPEAKER, RIGHT. >> YES, THANK YOU. >> BY THE WAY, THAT'S HER IN THE SLIDE UP ON THE TOP. >> SO I WILL TELL YOU A LITTLE BIT TODAY ABOUT THE ORIGINS OF TWO HUMAN MALARIA PARASITES. THIS IS SORT OF AN OUE OF MY TALK. I'M GOING TO GIVE A VERY BRIEF INTRODUCTION AND THEN TALK ABOUT PLASMODIUM INSPECTIONS AND COVER THE ORIGIN OF THE TWO PARASITES. TALK A LITTLE BIT ABOUT THE LIKELIHOOD OF ADDITIONAL PLASMODIUM ZOONOSIS AND TALK ABOUT THE AID OF HUMAN MALARIA. JUST TO BRING EVERYBODY ON THE SAME PAGE BECAUSE I HEARD THAT YOU HAVE VERY DIFFERENT BACKGROUNDS, YOU MUST KNOW THAT MALARIA IS CAUSED BY PARASITE OF THE VENOUS PLASMODIUM. THERE ARE PARASITES LISTED HERE. AND IT'S REALLY -- IN PARTICULAR AND -- ARE THE MAJOR KILLERS, BEING RAVAGING THROUGH AFRICA AND -- BEING THE MOST WIDE SPREAD MALARIA PARASITE ON THE GLOBE. SIZE MATTERS? WELL, I'M AN HIV VIROLOGIST AND WE HAD TO GET USED IT. IT HAS 14 CHROMOSOMES ABOUT 5,300 GENES. HUMAN GENOME IS BY THE HUNDRED FOLD LARGER, BACTERIA ARE SMALLER AND IT CAN BE VERY SMALL. AND WE STILL HAVEN'T FIGURED THAT ONE OUT SO THAT TELLS YOU HOW LONG WE'RE GOING TO WORK ON THIS. WHAT DOES THE EXTRA GENOME START A COMPLEX LIFE CYCLE. THERE'S THE [INDISCERNIBLE] HOST AND THE MOSQUITO HOST. I'M NOT GOING TO GO THROUGH THE DETAILS BECAUSE IT DOESN'T MATTER FOR MY TALK AND I'M SURE CAROLINA WILL COVER THE MOSQUITO BASE. THERE'S A LIVER STATE AND A BLOOD STATE AND THERE ARE [INDISCERNIBLE] THAT ARE BEING PICKED UP BY THE MOSQUITO AGAIN. THIS IS WHERE SEXUAL REPRODUCTION OCCURS ANDw3 THIS IS WHERE ASEXUAL REPRODUCTION OCCURS. THE POINT IS MALARIA'S A BAD THING, ALMOST TOP OF THE WORLD'S POPULATION IS AT RISK. THERE ARE NEW INFECTIONS EVERY YEAR. OVER A MILLION DEATHS PRIMARILY IN KIDS IN AFRICA. IT'S SOMETHING YOU DON'T WANT TO HAVE. THERE'S NO GOOD VACCINE YET, ALTHOUGH IT'S LOOKING MORE PROMISING. AND THERE'S WIDE SPREAD DRUG RESISTANCE SO THIS IS A PROBLEM. SO FOLLOW GENETICALLY WHERE DO THE HUMAN PARASITES FALL. THIS IS SEQUENCES OF A VARIETY OF DIFFERENT PARASITES. THERE ARE FOUR GROUPS, THERE'S A PRIMARY GROUP ONE, A PRIMARY GROUPc IS -- AND THE GROUP ONE ARE THE OTHER FOUR. AND IT WAS PARASITES FROM RODENTS AND FROM BIRDS AND [INDISCERNIBLE]. SO THIS IS IMPORTANT BECAUSE THIS LINEAGE HERE IS QUITE DIFFERENT FROM THE OTHER LINEAGE THAT INCLUDES THE HUMAN PARASITES AND ACTUALLY HAS BEEN SOME TIME AGO WAS SUPPOSED TO BE NAMED A DIFFERENT SUBGENUS. YOU WILL FIND THAT'S A HAY HELPFUL NOMENCLATURE AS I GO ON. SO THERE ARE A LOT OF PARASITES INFECTING PRIMATES IN ASIA. INCLUDING ORANGUTANS. THERE'S ALSO PLASMODIUM MAYOR SITES IN THE NEW WORLD IN THE AMERICAS. HOWEVER, IT SHOULD BE POINTED OUT THAT THESE ARE ACTUALLY REVERSE [INDISCERNIBLE]. THERE ARE A NUMBER OF NEW WORLD MONKEY SPECIES INFECTED WHICH IS CALLED PLASMODIUM BRAZILIAN BUT IT'S ACTUALLY MALARIA WAS BROUGHT TO THE NEW WORLD AND THEN JUMPED FROM HUMANS TO MONKEYS. PLASMODIUM INFECTS HOW MONKEYS AND SPIDER MONKEYS AND THIS REALLY WAS BROUGHT TO THE NEW WORLD BY HUMANS. IT'S IMPORTANT BECAUSE IT SHOWS YOU THAT THESE PARASITES CAN JUMP SPECIES. IN AFRICA, THERE ARE PARASITES IN MONKEYS BUT ALSO IN APES. ABOUT A HUNDRED YEARS AGO PEOPLE WERE STUDYING PLASMODIUM INFECTIONS IN AFRICA AND AT THE TIME THEY DIDN'T HAVE ANY GENETIC TOOLS BUT THEY LOOKED IN THE MICROSCOPE AND FOUND THE SAME THING THEY COULD FIND IN HUMANS. AND SO THEY GAVE THEM NAMES PLASMODIUM [INDISCERNIBLE] AND [INDISCERNIBLE] AND UNFORTUNATELY THERE ARE NO STORED SAMPLES THAT WOULD ALLOW US TODAY TO FIGURE OUT WHAT THEY ACTUALLY WERE. AND THEN THERE'S [INDISCERNIBLE] LEMURS, THEY HAVE THEIR OWN SPECIES. SO ONE THING THAT'S IMPORTANT THAT I WANT YOU TO TAKE HOME IS THAT THESE TWO GROUPS OF PLASMODIUM PARASITES ARE VERY DIFFERENT WITH RESPECT TO THEIR HOST SPECIES SPECIFICITY. THESE GUYS HERE ARE QUITE PROMISCUOUS. I TOLD YOU ABOUT THE HUMAN PARASITES GOING INTO NEW WORLD MONKEYS. BUT AS IT TURNS OUT, THAT PLASMODIUM IS A PARASITE THAT INFECTS AND ATTRACTS IN SOUTHEAST ASIA AND THAT JUMPS OVER TO HUMANS AND CAUSES QUITE A BIT OF MORBIDITY AND EVEN SOME MORTALITY IN THESE AREAS. AND IT'S I THINK STILL NOT SHOWN THAT THIS PARASITE CAN BE TRANSMITTED FROM HUMANS TO HUMANS. AND EACH CASE IT IS IN THE CLASS OF THE HUMAN [INDISCERNIBLE] ALSO HAS BEEN FOUND IN HUMANS ALTHOUGH IN THESE CASES THESE ARE LABORATORY TRANSMISSION. BUT STILL, IT SHOWS YOU THAT THIS GROUP WHAT WE CALL NON--- [INDISCERNIBLE] THIS IS NOT THE CASE BECAUSE A HUNDRED YEARS AGO PEOPLE WHO WERE STUDYING MALARIAS IN AFRICA WERE DOING EXPERIMENTS, SOMETHING THAT IS UNHEARD OF TODAY. BUT THEY SHOWED THAT WHILE THEY COULD TRANSMIT SOME OF THESE FROM APRILS TO HUMANS AND HUMANS TO APES THEY COULD NOT DO THAT WITH PARASITES INFECTING CHIMPANZEES. SO FOLLOWING THE RELATIONSHIP -- MM-MM, SOMEHOW MY -- THE ANIMATION DOESN'T WORK BUT IT'S OKAY. SO PEOPLE HAD ACTUALLY THOUGHT OVER COMING FROM THIS RELATIONSHIP IS THAT THEY SAW A CHIMPANZEE PARASITE AND A HUMAN PEER SITE AND THEY SAID WELL MAYBE THESE PARASITES HAVE [INDISCERNIBLE] WITH THE HOST AND HENCE WHEN HUMANS AND CHIMPS LIVED ABOUT SIX MILLION YEARS AGO MOST LIKELY THESE PARASITES HAVE BEEN IN THESE SPECIES FOR THIS LONG A PERIOD OF TIME. BECAUSE [INDISCERNIBLE] BECAUSE THERE ARE A LOT OF OTHER PARASITES HERE THAT FALL IN THE SAME GROUP PEOPLE SAID WELL THAT MUST HAVE BEEN ORIGINATED IN ASIA AND IT MUST HAVE BEEN CROSS SPECIES TRANSMISSION FROM AN ASIAN MONKEY PARASITE. AND BOTH OF THESE ARE, BOTH OF THESE HYPOTHESES ARE WRONG AND I'M GOING TO SHOW YOU WHY. IF THERE'S NOTHING ELSE FOR YOU TO TAKE HOME TODAY, TAKE THAT HOME. IT'S NOT OF ASIAN ORIGIN AND PLASMODIUM DOES NOT CURRY WELL. WHY DO ix5 KNOW THIS AND HOW DOES AN HIV PERSON GET INTO THE MALARIA FIELD. THE REASON IS WE'VE BEEN INTERESTED IN THE ORIGIN OF THE HUMAN APE VIRUS HIV-1 AND WE TRACED IT BACK TO CHIMPANZEES. AND THE WAY WE LOOKED AT IS NON-INVASIVELY. BECAUSE CHIMPANZEES ARE INVASIVE YOU CAN NOT STUDY THEM IN THE WILD BY ANNETTE TIEING THEM OR DOING ANYTHING INVASIVE SO THE ONLY THING LEFT TO STUDY IS A FECAL SAMPLE. WE'VE BEEN DOING THIS FOR THE PAST DECADE COLLECTING FECAL SAMPLES AND IT IS AMAZING WHAT YOU CAN GET OUT OF FECAL SAMPLES. FOR STARTERS, YOU CAN GET ANTI-BODIES. IF YOU HAVE AN INFECTION, YOU CAN DO MORE A SENSITIVE TYPE OF BLOOD AND YOU CAN FIND ANTIBODY REACTIVITY. BUT YOU CAN ALSO EXTRACT [INDISCERNIBLE] WHICH ALLOWS YOU TO EXTRACT RNA IN THE CASE OF HIV OR HOST DNA. SO YOU CAN GET FULL LENGTH AND PARTIAL SIV GENOMES, YOU CAN DO [INDISCERNIBLE] ANALYSIS, YOU CAN EVEN RECONSTRUCT INFECTIOUS MOLECULAR CONES. YOU CAN RESURRECT THE VIRUS FROM FECAL SAMPLES WHICH WAS DONE IN THE CASE OF THE GORILLA VIRUS. YOU CAN ALSO STUDY THE HOST. YOU CAN DO MITOCHONDRIAL DNA ANALYSIS AND YOU CAN VERIFY THE SPECIES AND SUBSPECIES AND YOU CAN ENUMERATE INDIVIDUALS WHICH ALLOWS YOU TO DO PREVALENCE DETERMINATION. SO WE'VE BEEN DOING THIS FOR HIV AND WE ACTUALLY TRACE IT BACK TO THE HIV-1 CHIMPANZEES AND [INDISCERNIBLE] TO CHIMP'S AND GORILLAS. WE DID THIS BACK IN 2006. BUT IN 2009, A PAPER WAS PUBLISHED BY A FRENCH GROUP. IN FACT WORKING WITH OUR COLLABORATORS AND WHAT THEY SHOWED YOU COULD ALSO DETECT PLASMODIUM SEQUENCES AND AT THAT POINT WE KNEW WE WERE IN BUSINESS BECAUSE WE HAD 10,000 IN THE FREEZER. SO IT WAS A NO BRAINER TO LOOK OF WHAT WE COULD FIND IN THESE SAMPLES. WHEN I WAS FIRST APPROACHED TO LOOK FOR THAT, THIS IS ACTUALLY RICHARD CARTER WHO HAS BEEN IN THE MALARIA FIELD FOR A LONG TIME, I SAID YOU WILL NEVER FIND PLASMODIUM DNA IN FECAL SAMPLES. WHY WOULD IT BE THERE. IT TURNS OUT I WAS DEAD WRONG AND THE REASON IT IS THERE IS BECAUSE OF THE LIVER STAGE AND WE JUST PUBLISHED A PAPER WITH RICHARD SHOWING A LIVER INFECTION ACTUALLY LEADS TO SECRETION OF PARASITE ACID INTO THE BILE AND FROM THE BILE INTO THE FECAL SAMPLE AND THAT'S WHY YOU GET PLASMODIUM SEQUENCES IN FECAL SAMPLES. IT TOOK SOME TIME TO FIGURE OUT. SO GIVEN THAT, WE WONDERED WHAT WE MIGHT FIND IN AIDS. WE KNEW FROM THE OLD STUDIES THAT THERE WERE EQUIVALENT OF ALL OF THE HUMAN PARASITES IN APES . SO THIS WAS THE CHIMP EQUIVALENT OF [INDISCERNIBLE] WAS THE CHIMP OR PEOPLE PLIERMLY LOOKED AT CHIMPANZEES BUT SOME ALSO DID IN GORILLAS -- DIFFERENTIATE THE EQUIVALENT OF [INDISCERNIBLE]. BUT ONLY RIGHT NOW IT MADE IT INTO THE MODERN DAY IN TERMS OF CULTURING THE PARASITE AND DOING THE GENETIC ANALYSIS NO SAMPLES ARE WERE LEFT FROM THESE DESCRIPTIONS WHICH WERE BASICALLY A DESCRIPTION OF [INDISCERNIBLE] AND ANALYSIS DONE BACK BY EDWARD IN CAMEROON AND THESE OTHER PEOPLE A HUNDRED YEARS AGO. THEY WERE INTERESTED IN HUMAN MALARIA AND THEY FIGURED WELL MAYBE OUR CLOSEST RELATIVES HAVE IT TOO AND SURE ENOUGH THEY DID. SO THE QUESTIONS WE WANTED TO ADDRESS GIVEN OUR BEEN REPOSITORIES, WE WANTED TO KNOW WHICH APE SPECIES AND SUBSPECIES ARE ACTUALLY INFECTED IN THE WILD WITH PLASMODIUM, HOW PREVALENT WAS THIS ON INFECTION HOW MANY DIFFERENT PLASMODIUM SPECIES WERE CIRCULATING AND WERE THE PARASITES OF APE ORIGIN. WE DID WHAT EVERYBODY ELSE DID WE TARGETED REYNOLDS OF THE PLASMODIUM MITOCHONDRIAL GENOME, AMPLIFIED BY A THOUSAND BASIS. THIS IS IN THE [INDISCERNIBLE] GENE. SOMEBODY TOOK OUR FECAL SAMPLES AND TOOK THE DNA AND SCREENED THE HELL OUT OF THEM. THE RESULTS WERE WE FOUND [INDISCERNIBLE] INFECTIONS ALL OVER THE PLACE. EVERY SINGLE SUBSPECIES OF WHICH THERE ARE FOUR [INDISCERNIBLE] EVERY SINGLE SUBSPECIES WAS U INFECTED. THEY COLLECTED SAMPLES IN YELLOW BEING THOSE THAT WERE ACTUALLY POSITIVE. INTERESTINGLY ENOUGH, CHALLENGE PAN SEE THERE'S A SECOND SPECIES [INDISCERNIBLE] WE DID NOT FIND ANY INFECTIONS IN THESE ANIMALS. THE RANGE IS SOUTH OF THE CONGO RIVER. WE HAVE SINCE EXPANDED INTO APES AND STILL HAVE NOT FOUND IT. GORILLAS, SIMILARLY THIS IS THE WESTERN GORILLA. THIS IS THE EASTERN GORILLA. LOTS OF INFECTIONS, HIGHLY PREVALENT IN WESTERN GORILLAS. WE DID NOT FIND INFECTIONS IN EASTERN GORILLAS BUT HAVE TO SAY WE ONLY LOOKED AT 146 FECAL SAMPLES THERE, WHEREAS WE HAD A LOT MORE, CLOSE TO 3,000 FECAL SAMPLES HERE. THE REASON BEING WHO COLLECT FROM THESE POPULATIONS SAID THERE ARE REBELS IN THE FOREST WITH MACHINE GUNS AND I SAID DON'T GO THERE. WE ONLY HAVE FEW SAMPLES. HOPEFULLY WHEN THE POLITICAL SITUATION IN THE DRC CALMS DOWN WE WILL GET MORE SAMPLES FROM THAT REGION. SO CHIMPANZEES AND WESTERN GORILLAS INFECTED AT A HIGH PREVALENCE. AND REMEMBER THIS IS A FECAL PREVALENCE. SO IT'S PROBABLY A VAST UNDER ESTIMATED OF WHAT IS ACTUALLY THERE. WE THEN PUT THE SEQUENCES INTO A COLLAGEN [INDISCERNIBLE] AND WE HAD A SURPRISE. INSTEAD OF THE TWO LINEAGES THAT WERE KNOWN BEFORE BEEN IN DEAR, THIS IS HUMAN [INDISCERNIBLE] WE ACTUALLY FOUND SIX MAJOR LINEAGES WHICH TRANSLATES INTO SIX SPECIES. IN RED IS MORE SPECIFIC FOR CHIMPANZEES, THREE OF THEM IN GREEN WERE SPECIFIC FOR GORILLAS. AND YOU SEE HERE WHERE HUMAN FALLS IT FALLS WITHIN THE RADIATION OF ONE OF THESE THREE GORILLA PARASITES. EVERY SINGLE HUMAN SEQUENCE COLESS BACK TO ONE COMMON ANCESTOR AND THAT'S FALLS WITHIN THE RADIATION OF THE GORILLA PARASITES AND THAT'S THERE WAS A GORILLA TO HUMAN TRANSMISSION THAT GENERATED HUMAN FECAL SYSTEM. SO THIS IS NOT NOW SCHEMATIC FORM WHAT WE FOUND AND WE'VE SINCE GIVEN THESE LINEAGES SPECIES A NAME. SOME PEOPLE OBJECT TO THAT BECAUSE GIVING A SPECIES NAME REQUIRES MORE PLASMA MORPHOLOGY AND OTHER THINGS. BUT BECAUSE WHILE APES ARE INACCESSIBLE THIS IS ALL WE CAN DO. THE REASON WE'–śU GIVEN THESE SPECIES NAMES SO THAT WEP REFER TO THEM, SO WE CAN DIFFERENTIATE WHAT WE CALL [INDISCERNIBLE] FROM CHIMPANZEES. REMEMBER THE -- THE CLOSEST RELATIVE OF WHAT WE CALL [INDISCERNIBLE] BECAUSE IT'S THE GORILLA PARASITE [INDISCERNIBLE] AND [INDISCERNIBLE] IS THE CLOSEST RELATIVE AND THE OTHER TWO FALL IN THE MIDDLE AND HERE ARE THE OTHER PARASITES. SO TAKE HOME AT THIS POINT GORILLAS ARE NATURALLY INFECTED WITH PRE-- SPECIES. IF INFECTION ARE HIGHLY PREVALENT, WIDELY DISTRIBUTED AND ALMOST ALWAYS COMPRISED OF MIXED PARASITE INFECTIONS. I DIDN'T SHOW YOU THAT BUT WE WOULD ALMOST ALWAYS FIND NOT JUST ONE BUT ALL THREE OF THE CHIMPS OR GORILLAS SPECIFIC PARASITES BUT NEVER THE CROSS SPECIES TRANSMISSION. AND THERE ARE REMARKABLY HOST SPECIFIC SO YOU MUST UNDERSTAND GORILLAS AND CHIMP'S LIVE IN THE SAME FOREST. THEY EAT THE SAME FRUIT TREES AND LIKELY BITTEN BY THE SAME MOSQUITOS BUT YET THEY HAVE VERY SPECIFIC PLASMODIUM PARASITES. FOLLOWING A SINGLE GORILLA -- ARE VERY RECENT HUMAN ORIGIN. I DIDN'T SHOW YOU THIS DATA. PEOPLE OF COURSE FIRST LOOKED IN CAPTIVITY WHAT THEY COULD FIND AND THEY COULD FIND HUMAN [INDISCERNIBLE] AND THEY SAID BONOBOS ARE THE ORIGIN BUT IT SO HAPPENS THAT THEY HAD PARASITES WITH HUMAN DRUG RESISTANCE LIMITATIONS IN THEM AND THEY WERE IN AN URBAN SANCTUARY. SO WHAT IT TELLS YOU THE BONOBOS ARE NOT [INDISCERNIBLE] THEY ARE SUSCEPTIBLE TO ION IF HE CANNIONS. IF YOU PUT THE RIGHT MOSQUITOS AROUND THEM THEY WILL BECOME INFESTED. WHY THEY'RE NOT INFECTED IN THE WILD IS A MYSTERY. THERE IS NO EVIDENCE OF HUMAN IN APES OR MONKEYS. SO WHAT ABOUT THE OTHER PLASMODIUM SPECIES. WHEN WE DID THE SURVEY, WE FOUND ALSO EVIDENCE OF APES RELATIVE OF [INDISCERNIBLE] MALARIA AND IN FACT THERE ARE OTHER LINEAGES HERE THAT I AM NOT SHOWING HERE THAT WOULD INDICATE THAT APES HAVE ADDITIONAL PLASMODIUM SPECIES. IT DOES NOT HAVE A COUNTERPART. NOW BECAUSE OF THIS, FINDING [INDISCERNIBLE] IN APES, THAT WAS OF INTEREST TO US AND IT WAS OF INTEREST TO US BECAUSE OFFICE EPIDEMIOLOGY OF [INDISCERNIBLE] SO THIS IS THE GLOBAL SPREAD OF FULL ZIP RULE AND [INDISCERNIBLE] PEOPLE HERE ARE NEGATIVE -- WHICH IS THE RECEPTOR. AND PEOPLE HAVE ALWAYS WONDERED WHY IS THIS. WHERE IS THE PRESSURE OF PEOPLE WHO HAVE DEVELOPED THIS GENETIC MUTATION. SO THIS IS THE ANTIGEN RECEPTOR [INDISCERNIBLE] IT HAPPENS TO BE THE RECEPTOR FOR [INDISCERNIBLE] IT'S INTERACTS WITH THE DUFFY BINDING PROTEIN AND IF YOU DON'T HAVE THE RECEPTOR YOU GENERALLY DO NOT GET INFECTED. THIS WAS ACTUALLY DISCOVERED HERE. IN HUMANS, HUMANS HAVE REALLY A FABULOUS MUTATION. THE MUTATION IS REGULATORY DOMAIN OFF THIS PROTEIN THAT A GATES EXPRESSION NOT IN GENERAL BUT IN ERYTHROCYTES, WHERE IT COUNTS. SO WE KNOW HUMAN [INDISCERNIBLE] WAS VIRTUALLY ABSENT FROM WEST CENTRAL AFRICA, YET THERE WAS A MYSTERY AND THAT MYSTERY HAS BEEN THERE FOR QUITE A NUMBER OF DECADES. A PERSON OF TRAVELERS THAT RETURN TO EUROPE OR TO THE UNITED STATES OR TO AUSTRALIA THAT HAVE VISIT THESE PARTS OF AFRICA COME BACK WITH MALARIA. WHEN YOU LOOK IT'S NOT FULL ZIP RULE -- PEOPLE TRAVEL TO OTHER PLACES AS WELL. CONSISTENTLY AND TO HAVE PAPERS WRITTEN ABOUT THESE UNUSUAL CASES OF TRAVELERS BRINGING BACK. IN ADDITION [INDISCERNIBLE] PUBLISHED A PAPER SOME TIME AGO WHERE HE LOOKED IN THE BLOOD OF INHABITANTS OF THE REPUBLICAN OF CONGO AND FOUND ANTI-BODIES.0 SO HOW DO THESE PEOPLE GET ANTI-BODIES IF IT WASN'T THERE. SO WE WERE WONDERING IF THERE'S A NATURAL RESERVOIR. WE DID THE SAME THING AS BEFORE. WE TARGETED THE PLASMODIUM MITOCHONDRIAL DNA. THIS TIME WE DIDN'T GO FOR THE [INDISCERNIBLE] BUT THE COX 1 GENE BECAUSE OF PRIMARY DESIGN AND OTHER THINGS AND WE SCREENED AGAIN. WHEN THE FECAL SAMPLES EXTRACTED DNA, THIS TIME WE SCREENED ABOUT 5,000 FECAL SAMPLES AND HERE ARE THE RESULTS. WE FOUND DEFINITELY PARASITES? CENTRAL, EASTERN AND WESTERN GORILLAS AND EASTERN GORILLAS. WE DIDN'T FIND IT IN BONOBOS AGAIN BUT THE PROBLEM HERE IT DOES NOT GROW TO THE SAME TITER IN THE BLOOD SO IT ENDS UP IN THE FECAL SAMPLE LOWER AMOUNTS. WE HAD TO SCREEN ON AVERAGE A HUNDRED FECAL SAMPLES TO FIND ONE POSITIVE. THAT'S A LOT OF FECAL SAMPLES. AND IT'S CLEARLY A TITER BUSINESS. SO WE GOT INFECTION RATES WHICH WERE MUCH LOWER THAN WHAT WE HAD SEEN FOR THE -- WHICH WERE 40-50% HERE WE'RE TALKING 4-8%. EVEN IN HUMANS PARASITE RATES ENDEMONSTRATEALLY INFECTED -- WE'RE CLEARLY IN THE TRANSMISSIBLE RANGE WITH THESE NUMBERS. AND IN FACT IN HUMANS A PARASITE RATE GREATER THAN 1% INDICATES TRANSMISSION. SO THIS IS AGAIN THE MAP. AGAIN YELLOW MEANS VIE VAC IS PRESENT SO YOU SEE THE SPREAD. THERE ARE A NUMBER OF NEGATIVE SITES BUT I THINK THAT'S BECAUSE WE MISSED IT BECAUSE WE DIDN'T HAVE ENOUGH SAMPLES TO SCREEN. WE FOUND IT IN AN UNUSUAL REGION WHERE THERE ARE CHIMPANZEES -- THEY ARE DIFFERENT THAN THEIR FOREIGN COUNTERPARTS BECAUSE THEY HAVE SUCH TOUGH ENVIRONMENTAL CONDITIONS TO DEAL WITH. SIX MONTHS OUT OF THE YEAR IT DOESN'T RAIN. THEY ARE BEHAVIORALLY DIFFERENT FROM THEIR FOREST COUNTERPARTS AND EVEN THERE WE FOUND VIVAX NOT JUST ONCE BUT FOUR TIMES. SO IT'S WIDE SPREAD. AGAIN NOTHING IN BONOBO AND I DON'T KNOW WHY. SO AT THAT POINT WE WANTED TO DO THE SAME THING, AMPHI GENES FROM THE MITOCHONDRIAL GENOME FROM THE FECAL GUILTY TO COMPARE THE APE VERSIONS TO THE HUMAN VERSIONS IN ORDER TO FIGURE OUT WHAT THE ORIGINrF WAS. BECAUSE OF THE LOW PARASITE LEVELS, ALTHOUGH WE STARTED OUT WITH THE DIAGNOSTIC FRAGMENTS IN THE MITOCHONDRIA GENOME, WE HAD ABOUT MAYBE SOME POSITIVE SAMPLES BUT WHEN WE TRIED TO AMPLIFY LARGER PIECES WHICH WOULD BE NECESSARY TO HAVE A DECENT ANALYSIS IT GOT PRETTY SLIM HERE. AND SO WE REALIZED WE HAD TO DO SOMETHING DIFFERENT. AND SO WE WENT AND LOOKED FOR AFRICAN SANCTUARY. AFRICAN SANCTUARIES ARE GREAT. THE PARENTS OF THESE WERE [INDISCERNIBLE] THEY END UP IN THESE SANCTUARIES. THEY ARE IN CAPTIVITY. THERE IS A FENCE FENCING THEM IN BUT THE HABITAT IS IMMEDIATELY ADJACENT TO THE HABITAT OF WILD SO THEY GET BITTEN BY THE SAME MOSQUITO. YES, YOU CAN DO THE ANNUAL HEALTH EXAM AND YOU CAN DRAW BLOOD FROM THEM. THE REASON WE GOT INVOLVES WITH THE SANCTUARIES, WHEN WE GET THEM ANTI-MALARIAL DRUGS THEY GET BETTER. THAT'S WHY WE START TO SCREEN THESE APES AND THAT'S WHY WE HAD ACCESS TO BLOOD SAMPLES. THE SEQUENCES ARE COLOR CODED. HERE ARE THE CENTRAL, NIGERIAN EASTERN CHIMPS AND WESTERN GORILLAS. YOU SEE WHERE THE HUMAN VEY -- VIVAX STRAINS COME FROM AND THEY FORM TO A SINGLE COMMON ANCESTOR WHICH FALLS WITHIN THE RADIATION OF THE APE PARASITE. AND THIS IS TRUE FOR THE MITOCHONDRIAL GENOME, IT'S TRUE FOR A NUCLEAR GENE LBH, SAME THING. AND IT'S TRUE FOR A FECAL CLASS GENE CLCC. HERE THEY ARE ALL FEWER SEQUENCES AVAILABLE BUT THE SAME STORY. ALL THE HUMANS SEQUENCES FORM THE SINGLE LINEAGE WITHIN THE APE RADIATION. WHEN WE LOOK AT DIVERSITY COMPARING HUMAN VIVAX AND APE VIVAX WE ALWAYS SAW THAT THE APE PARASITES WERE MORE DIVERSE WHICH IS WHAT THEY WOULD EXPECT FROM A RESERVOIR TESTED. IT'S DIFFERENT FROM 1.4 TIMES MORE IN THE MITOCHONDRIAL GENE UP TO 50 IN SOME REGIONS. SO [INDISCERNIBLE] NOW GIVEN THAT VIVAX IS FROM THE MORE PROMISCUOUS LINEAGE WE WANTED TO BE SURE WE DIDN'T MISS SOMETHING IN THE MUST NOT KEEPS SO WE DID LOOK AT A THOUSAND BLOOD SAMPLES FROM [INDISCERNIBLE] SAMPLES, THESE ARE MONKEYS THAT ARE SOLD AT MEAT MARKETS FOR MEAT AND OUR COLLABORATOR HAS DRAWN THESE BLOOD SAMPLES, THEY ARE ALL WAY BACK WHEN WE WERE STUDYING HIV SO THESE CAME HANDY. AGAIN WE LOOKED AT 16 DIFFERENT SPECIES AT 11 DIFFERENT LOCATIONS AT THESE SITES AND WE FOUND NO EVIDENCE FOR VIVAX. BUT ALL SAMPLES HAVE [INDISCERNIBLE] WHICH IS A KNOWN MONKEY PARASITE. WE ALSO LOOKED AT THE DARK GENES, THE PROMOTER REGION AND THE GENE ITSELF. BECAUSE WE WANTED TO KNOW WHETHER THE BONOBOS HAD A NEGATIVE GENETIC MUTATION. YOU CAN'T SEE IT BUT THIS IS THE DUFFY MUTATION BUT ALL THE APES WERE WILD TYPE. THEY DIDN'T LOOK LIKE DUFFY MUTATION. THERE'S A [INDISCERNIBLE] WHICH IS ASSOCIATED WITH POTENTIALLY PROTECTIVE ALLELE IN HUMANS. AND AGAIN ALL THE REST WERE WILD TYPE. THEY HAD CHANGES SO I CAN'T EXCLUDE THE POSSIBILITY OF A DUFFY-LIKE CHANGE BUT AT LEAST THE HUMAN POSITIONS WERE WILD TYPE. SO I'VE SHOWN YOU THIS NOW. THIS IS HUMAN [INDISCERNIBLE] HERE'S HUMAN FAL [INDISCERNIBLE] WHAT WOULD YOU CONCLUDE ABOUT THE ORIGIN OF THE HUMAN VIVAX. WOULD YOU SAY THIS IS SINGLE APE TO HUMAN TRANSMISSION? NO. WHY NOT? BECAUSE OF THIS. NO SPECIES SPECIFICITY BETWEEN THE CHIMP AND THE GORILLAS. STRICT SPECIES SPECIFICITY. SO CLEARLY THE SPARE SITES GO FROM GORILLAS TO CHIMPS AND CHIMP'S TO GORILLAS AND MOST LIKELY AT SOME POINT IN THE PAST WENT FROM GORILLAS TO CHIMPS TO HUMANSdTO GORILLAS TO CHIMPS. IN OTHER WORDS THERE WAS AN ANCIENT STOCK OF VIVAX THAT WAS PROMISCUOUS THAT WENT AROUND ALL OF THESE DIFFERENT SPECIES. AND THE HUMANS DEVELOPED THE DUFFY NEGATIVE PHENOTYPE AND IN THE HUMAN POPULATION. WHAT YOU'RE LOOKING AT HERE IS ONE PARASITE THAT ESCAPED. OUT OF AFRICA. AND NOW IS CIRCULATING IN GLOBAL POPULATIONS. SO VIVAX DOES NOT DIVIDE INTO CHIMP AND SPECIFIC GORILLA LINEAGES. THIS IS THE ANCESTRAL STOCK I JUST TALKED ABOUT THAT WAS PROBABLY PRESENT IN ALL APE SPECIES UNTIL THE DUFFY NEGATIVE MUTATIONS AROSE AND STARTED TO SPREAD. AND DOES EXTEND HUMAN VIVAX REPRESENTS A BOTTLENECK IMAGE THAT IS SPREADING OUT OF AFRICA K SAME THING IN GORILLAS BUT NOT BONOBOS ARE [INDISCERNIBLE] THERE'S NO EVIDENCE OF VIVAX INFECTIONS IN OTHER AFRICAN PRIMATES. THERE'S NO EVIDENCE OF A DUFFY NEGATIVE PHENOTYPE OR SELECTION OF A PROTECTIVE ALLELE IN WILD APES ALTHOUGH I CANNOT EXCLUDE IF THERE ARE OTHER MUTATIONS THAT COULD CHANGES THE [INDISCERNIBLE] HUMAN P VIVAX AROSE FROM WITHIN A PLASMODIUM SPECIES THAT INFECTED HUMAN CHIMPS AND GORILLAS. THIS PROVIDES A LONG-SOUGHT EXPLANATION FOR THE SELECTION OF DUFFY NEGATIVE PHENOTYPE IN WEST AFRICA. THERE WAS THE TRANSMISSION THERE WAS THE REASON TO DEVELOP A DUFFY NEGATIVE PHENOTYPE. SO SINCE WAY NOW THE WHOLE SUBGENUS WITH SIX DIFFERENT APE LINEAGES, HUMAN AND APE VIVAX [INDISCERNIBLE] OCCURRING AREAS WHERE ION FECTED APES AND HUMANS LIVED IN CLOSE PROXIMITY -- ESPECIALLY HUMANS THAT WOULD LIVE IN VILLAGES CLOSE TO THE NATURAL TERRAIN OF GORILLAS WITH HIGH PREVALENCE OF LAUGH RAIN ANNUAL AND SO WE HAD THOSE PEOPLE. THE FIRST THING, THIS IS A VIEW OF ONE OF THESE WHICH SHOWS YOU THAT PEOPLE REALLY DO LIVE AND SLEEP. SO THE FIRST QUESTION WE ASK IS MOST SEQUENCES AT THIS POINT HAD COME FROM NOT NECESSARILY FROM WEST CENTRAL AFRICA. MAYBE WE MISSED THEM OR SOMETHING BECAUSE IN HIV THERE ARE FOUR TRANSMISSIONS AND IT'S NM -- WE THOUGHT WE JUST MISSED IT. SO WE LOOKED AT ALMOST 15 NEW INFECTIONS BUT EVERY SINGLE ONE FELL WITHIN THAT LINEAGE OF HUMAN, THERE WAS NOŽ ANY ADDITIONAL CROSS SPECIES. WHICH IS ODD BECAUSE WHEN SOMETHING DUMPS IT WORKS TO DUMP MORE THAN ONCE UNLESS THERE'S SOMETHING REALLY UNUSUAL GOING ON. WE ALSO DID SOME PYROSEQUENCING BECAUSE YOU ALSO GET INFECTED WITH THE AIDS VERSION BUT THE AIDS VERSION AREN'T ADAPTED TO HUMANS AND HENCE THE TITERS ARE LOWER AND WE MISSED IT. SO WE DID SEQUENCING TO SEE WHETHER WE WOULD FIND IT. AND SO IN ALMOST 450 HUMAN SAMPLES WE LOOKED AND ALL WE WOULD FIND IS THE INFECTION WITH HUMAN [INDISCERNIBLE] WE DID NOT FIND ANY AIDS. IF WE LOOKED IN FECAL SAMPLES WITH THE SAME PRIMARIES WE DID FIND ALL OF THESE DIFFERENT SPECIES. IT WASN'T BECAUSE THE PRIMARIES WEREN'T WORKING IT'S BECAUSE THEY WEREN'T INFECTED WITH AIDS LAUGH RAIN YEAH PARASITES. YES, THE SAME GROUP WHO DISCOVERED THE PLASMODIUM SEQUENCES FROM FECAL SAMPLES ALSO LOOKED AT VIVAX AND THIS IS THE SAME THING WE DID EXCEPT THEY DIDN'T DO IT IN THOUSANDS OF SAMPLES. THEY WENT TO [INDISCERNIBLE] THEY WERE IN SANCTUARIES AND THEY FOUND APE VIVAX AND THEY LOOKED IN [INDISCERNIBLE] FROM THESE REGIONS. THEY PUT OUT TRAPS CLOSE TO WHERE THE APES WERE OVER THIS PERIOD OF A YEAR A COUPLE DIFFERENT POSITIVE AND SHOWED THEY WERE INFECTED WITH THE AIDS PARASITE. THEN THEY FOUND ONE TRAVEL, ONE FRENCH FORESTER WHO HAD WORKED IN A FOREST IN THE CENTRAL AFRICAN REPUBLICAN FOR TWO WEEKS, RETURNED BACK TO FRANCE, HAS MALARIA, LOOKED AT IT, LOOKS LIKE VIVAX. BUT WHEN THE SEQUENCE WAS DETERMINED, IT WAS AN APE VERSION NOT THE HUMAN VERSION. BECAUSE WE HAD SO MANY SEQUENCES WE PUT THAT SEQUENCE IN OURS AND HERE'S THE HUMAN VIVAX AND HERE'S THE TRAVEL. CLEARLY THE TRAVEL PICKED UP AN APE PARASITE. BOTTOM LINE PEOPLE IN AFRICA THEY DON'T HAVE DUFFY THEY GET INFECTED IF YOU HAVE ONE AND IT GOES TO THE RIGHT PLACE COMES BACK WITH AN AIDS INFECTION. SO APE VIVAX CLEARLY HAS THE POTENTIAL TO CAUSE THE INFECTION. AND THIS IS 100% IN LINE WHERE THE PEOPLE HAVE DESCRIBED A HUNDRED YEARS AGO ABOUT EXPERIMENTAL TRANSMISSIONS OF AT THE TIME FROM APES TO HUMANS WHICH WORK AND YET THE DIFFERENCE FROM CHIMP'S TO HUMANS FAILED. SO THIS IS EXACTLY WHAT YOU WOULD EXPECT. SO TAKE HOME POINTS WAS INFECTED WITH PARASITES THAT DO NOT SERVE AS A RECURRING SOURCE OF HUMAN MALARIA. AND SO IT REALLY LEAVES US HANGING THERE WHY IT WAS DIFFERENT FROM GORILLAS IN THE FIRST PLACE. MAYBE CAROLINA WILL HAVE AN ANSWER ONE OF THESE DAYS. SHE CERTAINLY HAS AN IDEA. HUMANS ARE SAW SENTABLE TO INFECTION WITH VIVAX SO THE QUESTION IS WHAT WOULD HAPPEN IF YOU BRING IN AN INFLUX OF DUFFY POSITIVE PEOPLE INTO AREAS WHERE VIVAX IS ENDEMIC AND EVEN PERHAPS EXPRESS IT BACK OUT TO PLACES IN ASIA WHERE VIVAX IS NORMALLY TRANSMITTED. VIVAX IS MORE DIVERSE AND PERHAPS MORE VERSATILE. SO THIS IS SOMETHING TO WATCH. SO HOW OLD IS HUMAN MALARIA. AND THERE I HAVE TO SPECULATE. THERE ARE TWO RULES OF THOUGHT. SOME PEOPLE THINK IT'S VERY ANCIENT, HUNDREDS OF THOUSANDS OF YEARS OLD. OTHER PEOPLE THINK IT'S RECENT, LESS THAN 10,000 YEARS OLD. PEOPLE THAT THINK MALARIA IN HUMANS IS ANCIENT BASE THEIR CONCLUSIONS ON MOLECULAR THOUGHT. AND ALSO, ASSUMPTIONS THAT CERTAIN PARASITES [INDISCERNIBLE] AND THEN THEY USE EVOLUTIONARY RATES NOT FROM PLASMODIUM BECAUSE THAT'S NOT KNOWN BUT FROM OTHER SPECIES WHO TRIED TO CALCULATE THAT. PEOPLE WHO DON'T BELIEVE THAT MALARIA'S ANCIENT BASED IT ON THE PARASITE BIOLOGY. THEY HAVE OBSERVED SOME OF THE GENETIC MUTATIONS IN HUMANS HAVE NOT YET SPREAD THE FIXATION. THAT A GROUP OF [INDISCERNIBLE] IS LESS THAN 10,000 YEARS OLD AND THAT DUFFY EMERGED AROUND 30,000 YEARS AGO. SO THAT IS NOT COMPATIBLE WITH HUNDREDS OF THOUSANDS OF YEARS OF INFECTION´ ALSO THERE'S DATA THAT REALLY HAS A MAJOR EVENT ABOUT I THINK A TO 10,000 YEARS AGO WHEN HUMAN HUNTER GATHERERS GOT TOGETHER THERE WERE POOLS OF WATER AND STUFF LIKE THAT. SO IF YOU BELIEVE THE FIRST GROUP OF PEOPLE YOU CAN SIMPLY SAY IN THIS CASE, IT'S A PRECURSOR OF [INDISCERNIBLE] IN GORILLAS AND CHEM'S SO ROUGHLY SIX MILLION YEARS. IF YOU ASSUME THAT THEN YOU COME OUT WITH THE COMMON ANSWERS IN THE TO TO 300,000 YEARS AGO. BUT YOU CAN ALSO CALCULATE THIS DIFFERENTLY BASED ON THE MUTATION RATE OF PLASMODIUM WHICH OF COURSE HAS TO BE EVALUATED IN VITRO AND THE NUMBER OF CYCLES WAS IN A LIFE CYCLE. AND SO THESE ARE SOME OF THE ENVELOPE CALCULATIONS YOU HAVE TO ACCOUNT FOR MOSQUITOS ON THE LIVER AND BLOOD BUT PROBABLY THERE ARE ABOUT YOU KNOW ABOUT 500 REPLICATIONS PROBABLY NOT MORE AND POSSIBLY LESS. WHEN YOU PLUG ALL THAT IN, YOU COME UP WITH AN ESTIMATE THAT THE LAST COMMON ANSWERS COULD HAVE EASILY EXISTED 4,800 YEARS AGO. DON'T TAKE THAT NUMBER TOO SERIOUSLY, IT JUST MEANS THAT THIS WOULD BE MORE CONSISTENT WITH THE MORE RECENT ORIGIN OF HUMAN MALARIA. SO THE DIVERSITY AMONG DIFFERENT STRENGTHS CAN EASILY BE EXPLAINED BY DESCENT FROM ANN ZOSTER THAT EXISTED WITHIN THE LAST 10 YEARS. IT COULD BE OLDER BUT BECAUSE OF THE HYPNO ST. STAGE THE NUMBER OF GENOME REPLICATES IS DIFFICULT TO ESTIMATE AND SO MAYBE THE AGE OF THE DUFFY MUTATION WHICH HAS BEEN ESTIMATED ANYWHERE FROM 10 TO 30,000 YEARS AGO PROBABLY A BETTER CALIBRATION POINT OF THE TIME WHEN THAT LAST COMMON ANSWER COSMOPOLITAN VIVAX EMERGED. AND USING MUTATION RATES SEEM MORE CREDIBLE THAN MOLECULAR CLOCK ESTIMATES WHICH ARE NOTORIOUS FOR GIVING THE WRONG RESULTS ESPECIALLY WE TRIED TO DO THE SAME THING. THE FURTHER YOU GO BACK IN TIME THE LESS ACCURATE THESE CALCULATIONS ARE. FUTURE STUDIES VERY BRIEFLY. WE REALLY WOULD LIKE TO KNOW WHAT HAPPENED AND WHY THE GORILLAS PRECURSOR OF FALCIPARUM OVER COME THIS AND WE DO NOT HAVE AN ANSWER YET. BUT IT IS KNOWN THAT THE FORM HAS MANY SURFACE RECEPTORS THAT INTERACT WITH LIGANDS ON THE ERYTHROCYTES AND THEY ARE NOT ABSOLUTELY REQUIRED BUT ONE HAD BEEN RECENTLY FOUND BY THE SANGER GROUP TO BE ABSOLUTELY ESSENTIAL AND THAT'S CALLED THE RECEPTOR'S CALLED [INDISCERNIBLE] AND IT INTERACTS WITH THE RH5 PROTEIN ON THE PARASITES. AND SO BECAUSE IT IS SUCH AN ESSENTIAL INTERACTION, WE WERE INTERESTED IN WHAT THE EQUIVALENT OF OUR APE PARASITES WOULD LOOKfL„g LIKE. SO WE AMPLIFIED THE EQUIVALENT BECAUSE WHEN YOU TARGET SOMETHING YOU CAN ACTUALLY GET IT OUT. AND INTERESTINGLY ENOUGH, THIS IS WHAT WE GOT. AND THIS IS WHAT I SHOWED YOU BEFORE AND WHAT WE CAN SEE HERE IS THAT THE ORDER OF THE PARASITES IS NOT RIGHT. WHAT WE COULD SEE FALCIPARUM RIGHT NOW IS HERE. NO FULL ZIP RULE [INDISCERNIBLE] DOWN HERE. SO WITH WHAT THAT SUGGESTS TO US IS A HORIZONTAL GENE TRANSFER THAT WENT FROM WHAT WE CALL PLASMODIUM ANOTHER GORILLA PARASITE INTO THE PRECURSOR OF HUMAN FALCIPARUM. WE HAVE GENOMIC INFERENCE [INDISCERNIBLE] HAVING TO DO WITH THE TRANSMISSION WE DON'T KNOW BUT IT SURELY IS ODD. THAT'S THE ONE THING THAT'S ABSOLUTELY ESSENTIAL ALSO IS INVOLVED IN THIS HORIZONTAL TRANSFER. SO HOW CAN WE GET THE GENOMES OF THESE PARASITES. THE BLOOD WE GET FROM THE SANCTUARY, THESE ARE INDIVIDUALS BY AND LARGE THE PARASITE LOAD IS VERY LOW. WE CANNOT DO ANY WORK THERE WHICH MEANS WE HAVE WHOLE BLOOD USUALLY AND THAT'S WHAT WE HAVE TO WORK WITH. THAT USUALLY DOESN'T WORK IN WHOLE GENOME SEQUENCING. SO MY LAB CAME UP WITH WHAT IS CALLED SELECTIVE WHOLE GENOME AMPLIFICATION BASED ON THE FACT THAT THEY HAVE UNIQUE MUTATIONAL BIASES SO YOU [INDISCERNIBLE] AND YOU CAN THEN USE A 529 POLYMERASE THAT BASICALLY SPECIFICALLY OR ALMOST SPECIFICALLY AMPLIFIES THE PLASMODIUM AND LEAVES THE CHIMPS CONTAMINATING DNA ALONE. SO WE'VE DONE THAT AND WE'VE SEEN 30,000 ENRICHMENT AND THIS IS JUST TO SHOW THAT WE CAN DO IT. IT WAS GREAT WITH BLOOD SAMPLES, IT DOESN'T WORK GREAT WITH FECAL SAMPLES SO THERE'S STILL DEVELOPMENT OF WORK WE NEED TO DO. THIS IS THE END OF THE CHROMOSOME SEQUENCE AND WE GOT IT THERE AND WE HAVE VERY FEW GAPS IN THE SEQUENCE. SO WE WILL HAVE A WAY TO LOOK AT THE GENOMES OF THESE PARASITES AND TO SEE THAT THEY HAVE TO HAVE THE BLUEPRINT OF WHAT HAPPENED TO SEE WHY THEY JUMPED INTO HUMANS ONLY ONCE. SO MY CONCLUSION, THREE MAJOR HUMAN PATHOGENS ORIGINATED IN APES . OBVIOUSLY THE TRANSMISSION HURDLE FOR APE PATHOGENS IS LOWER, THAT'S NOT SUPER SIZING BECAUSE THESE PARASITES AND VIRUSES AND PATHOGENS HAVE TO INTERACT WITH HOST PROTEINS AND THE CLOSER THEY ARE THE EASIER IT IS FOR THEM. STILL ONLY A SMALL SUBSET HAS ACTUALLY SUCCESSFULLY COLONIZED HUMANS AND THAT TELLS YOU THERE ARE RESTRICTIONS ACROSS SPECIES TRANSMISSION. UNDERSTANDING THEM IS VERY IMPORTANT BECAUSE IT COULD GIVE YOU A GLIMPSE AT WHAT TO DO AND COUNTERACT THESE INFECTIONS. HOPEFULLY I CONVINCED THAT NON-HONEY VASIVE STUDIES OF WILD APES HAVE BEEN ENFORCED AND THERE'S VALUE OF STUDYING APE PRE CURSERS OF HUMAN PATHOGENS AND THEIR NATURAL HOSTS. ALTHOUGH IT DOESN'T IMMEDIATELY GIVE YOU CLINICAL BENEFIT FOR HUMANS. THESE ARE ALL MY COLLABORATORS PLUS THE PEOPLE IN MY LAB. I MENTIONED [INDISCERNIBLE] AND HER TEAM IN CAMEROON. MY FRIEND [INDISCERNIBLE] AND [INDISCERNIBLE] WHO DOES THE ANALYSIS FOR US. LOTS OF PEOPLE WHO GAVE US HUMANS AND EVEN MORE PEOPLE WHO GAVE US FECAL SAMPLES OF COURSE. THAT'S IT. [APPLAUSE] >> THANK YOU VERY MUCH. WE HAVE TIME FOR SOME QUESTIONS. I ONLY ASK THAT YOU EITHER SPEAK LOUDLY OR USE THE MICROPHONE SO PEOPLE WATCHING CAN HEAR THE QUESTION. LET ME READ OFF. WHY IS THERE NO FALCIPARUM IN SOUTH AMERICA. >> THERE IS, THERE IS PLENTY. THERE IS PLENTY OF FALCIPARUM IN SOUTH AMERICA. >> NOT ON YOUR SLIDE. >> NO, NO, THERE WAS. >> I DIDN'T SEE ANY BLUE THERE. >> NO, THERE IS. >> I THOUGHT THERE WAS THAT'S WHY. >> NO, THERE IS. THE ONLY BIG HOLE IS, HERE, SORRY. SO I MEAN SO HERE IS THE VIVAX AND HERE IS THE FALCIPARUM. >> OKAY.Sy– >> SO HOW DO YOU THINK THE [INDISCERNIBLE] COULD THEY EVOLVE WITH THE CHIMP AND THE GORILLA EVOLUTION? >> THAT'S ONE POTENTIAL EXPLANATION BUT I HAVE NO EVIDENCE TO EITHER CONFIRM OR REFUTE IT. YOU COULD EXPLAIN IT THAT WAY. HOW THESE PARASITES GOT INTO THE APES I REALLY DON'T KNOW. THIS IS ALSO FOR THE [INDISCERNIBLE] I DON'T WANT TO MAKE ANY ASSUMPTIONS BECAUSE THEN I'M BACK TO YOU KNOW HOW FALCIPARUM EVOLVED. WHAT IS MORE INTERESTING IS WHY THERE IS THIS SMASHES SPECIFICITY WHEN THEY LIVE IN THE SAME PLACE AND HAVE PRESUMABLY THE SAME HOST. A BIG HOLE GAP IN KNOWLEDGE RIGHT NOW IS THE MOSQUITO PAR BECAUSE WE NEED TO FIGURE OUT WHICH SPECIES ARE TRANSMITTING THESE PARASITES AMONG APES AND HOW THEY WORK BECAUSE THAT COULD EXPLAIN A NUMBER OF THESE THINGS. >> [INDISCERNIBLE]. >> SORRY. >> THANK YOU. IF YOU THINK THERE'S A [INDISCERNIBLE] GENE TRANSFERS DO YOU KNOW HOW THAT WOULD HAVE OCCURRED IN THE MOSQUITO. >> I HAVE NO IDEA HOW THIS WORKS. ACTUALLY THERE'S NO MECHANISM I COULD POINT TO THAT WOULD MAKE SENSE. I JUST DON'T KNOW. >> FOR A LONG TIME THERE WAS A NOTION THAT SICKLE CELL DISEASE HAS SOME PRESSURE FOR TRANSMISSION OF FALCIPARUM. WHERE DOES THAT STORY STAND TODAY? >> I THINK SICKLE CELL DISEASES IS PARTIALLY PROTECTIVE OF THE PATHOGENIC CONSEQUENCES OF INFECTION. I DON'T KNOW WHETHER IT HAS ANYTHING TO DO WITH TRANSMISSION. I DON'T KNOW. >> MY UNDERSTANDING IS THAT [INDISCERNIBLE] PROTECTS YOU FROM THE SEVERE FORM, PROTECTS CHILDREN FROM SEVERE MALARIA BUT YOU STILL GET INFECTED BUT IT'S NOT LETHAL. >> SO IS THAT ALSO TRUE FOR THE [INDISCERNIBLE] TRAITS, SAME THING TYPE. >> YES. >> THAT'S EVOLUTIONARILY PRESERVED ESSENTIALLY BY REDUCING. >> YES. I THINK THOSE WHO ARE HOMOZYGOUS IS REALLY BAD BECAUSE YOU'RE SICK BUT YOUR HEATHER ZYGOUS AND YOU'RE HEALTHY YOU'RE PROTECTED FROM SEVERE MALARIA SO THEY HAVE STAYED IN THE POPULATION IN AFRICA. IT'S ANOTHER EVIDENCE OF STRONG CONNECTION OF HUMAN POPULATIONS. >> AND THE DUFFY, THERE YOU WANT TO BE HOMOZYGOUS BECAUSE YOU HAVE TO NOT LEAVE THE DUFFY ANTIGEN ON THE ERYTHROCYTE. BUT THAT'S A VERY UNUSUAL MUTATION. >> ARE PRIMATES ILL WHEN THEY HAVE INFECTION. >> I THINK SO. SOMETIMES. BECAUSE THE REASON WE GOT INTO THIS IS BECAUSE THE SAGE CHERRY VETERINARIAN WOULD DESCRIBE CASES THAT WOULD LIKE LIKE CEREBRAL MALARIA. THIS IS UP IN THE FOREST, THERE'S NO INFRASTRUCTURE, EVEN THE BLOOD SMEARS. BUT WHEN SHE GAVE MALARIAL DRUGS, CHIMPS WOULD GET BETTER AND THAT HAPPENED THREE OR – FOUR TIMES. THAT'S WHY [INDISCERNIBLE] IT WAS GOING ON IN MY APES. WE SEE MORE INFECTION THAN DISEASE. THAT COULD BE A SIMILAR STORY WITH HUMANS WHERE MOST PEOPLE HAVE IMMUNE SYSTEMS, IMMUNE RESPONSE THAT IS PARTIALLY PROTECTED. AND IT ONLY HITS THE KIDS AND IT HITS THE PREGNANT WOMEN. >> BUT IN HUMANS [INDISCERNIBLE] WHAT ABOUT IN THE PRIMATES. >> WE DON'T REALLY KNOW. BECAUSE THESE APES ARE FIVE, SIX, SEVEN YEARS OLD. >> YOU TOUCHED ON IT A LITTLE BIT. DID APES ALSO SUFFER FROM PREGNANCY ASSOCIATED BY MALARIA INFECTIONS. DO APES ALSO SUFFER FROM PREGNANCY ASSOCIATED -- >> I DO NOT KNOW BECAUSE ONE THING THEY DON'T DO IN THE SANCTUARY THEY DON'T BREED THEM. THEY DON'T NEED MORE, THEY CAN HARDLY FEED THE ONES THEY HAVE. >> THANK YOU FOR YOUR PRESENTATION. I WOULD LIKE TO KNOW WHAT THE LEVEL OF DRUG RESISTANCE IN APES AS COMPARED TO HUMANS. >> WE HAVEN'T LOOKED YET. WE'RE JUST BEGINNING TO GET THE WHOLE GENOME. WE LOOK AT THE GENES THAT ARE INVOLVED IN THAT. WE HAVEN'T LOOKED YET. >> THANK YOU. >> I WOULD EXPECT NONE. >> IS THERE A WAY OF SEEKING THE VIRUS PRESENT [INDISCERNIBLE] FOR INSTANCE FROM THE MOTHER TO THE [INDISCERNIBLE] PREGNANCY. >> IN APES I HAVE NO IDEA BUT I THINK, I WOULD SPECULATE THAT APES LAUGH YAIN YEAH AND EVEN VIVAX INFECTIONS ARE VERY SIMILAR TO THE HUMAN COUNTERPARTS IN THESE THINGS BUT I HAVE NO KNOWLEDGE WHAT HAPPENS IN APES . >> DO YOU KNOW OTHER MUTATIONS THAT AFFECT EITHER THE CAPACITY FOR THE HEPATIC PHASE OR I MEAN FROM WHAT I'VE READ OF AT THIS TIME, THE INTRAHEPATIC PHASE PARTICIPATES TO A CONSIDERABLE EX2E7B9 IN THE INDO SAIDIC CYCLE AND THERE ARE HUMAN DISEASES THAT AWE FACT THAT CYCLE. IS THERE ANY EVIDENCE THAT THAT PLAYS A PART IN MALARIA. >> WE WOULD HAVE TO SEQUENCE THE GENOMES OF ALL THESE DIFFERENT APES TO SEE WHETHER THERE ARE SIMILAR CHANGES IN THEIR GENES AND WE HAVEN'T DONE THAT. >> OKAY. WELL LISTEN, THANK YOU IMMENSELY AND WE WILL PROCEED NOW T. [APPLAUSE] [„ >> GOOD AFTERNOON. THERE'S A LOT OF INTEREST IN UNDERSTANDING THE INTERACTIONS YOU'VE SEEN WITH [INDISCERNIBLE] TRANSMISSION. I WOULD LIKE TO START WITH JUST A COUPLE SLIDES TELLING YOU A LITTLE BIT ABOUT WHAT IS IT ABOUT MALARIA THAT MAKES HUMANS SO SICK [INDISCERNIBLE] I WILL BRIEFLY TOUCH ON THAT. THEN MOST OF MY TALK WILL BE EITHER TRYING TO GIVE YOU A SENSE WHAT HAPPENS WITH [INDISCERNIBLE] MALARIA, HOW THE MOSQUITO IMMUNE SYSTEM CONTAINS THIS INFECTION AND VERY POWERFUL ANTI-PLASMODIUM RESPONSES AND WE'LL SEE HOW IT LIMITS INFECTION. AND EVEN OF COURSE MOSQUITO [INDISCERNIBLE] THIS IS SOMETHING THAT HAS ALWAYS PUZZLED ME. I THINK THE ANSWER IS PLASMODIUM [INDISCERNIBLE] IS NOT A NORMAL PART [INDISCERNIBLE] HAS LEARNED TO MAKE IT SO INCONVENIENCABLE GIVEN THE MOSQUITO IMMUNE SYSTEM. AND THE EVIDE THESE AND THE GENES FROM THE MECHANISMS THAT MAKE THESE. AS I MENTIONED BEFORE, MALARIA IS A VERY SEVERE DISEASE THAT AFFECTS 40% OF THE WORLD POPULATION, 90% OF THE [INDISCERNIBLE] AND I'M TALKING ABOUT MALARIA IN THIS CASE. SO [INDISCERNIBLE] THEY INFECT THE LIVER AND THEN THERE'S INFECTION OF THE MULTIPLICATION IN RED BLOOD CELLS AND BASICALLY MOST OF THE SEVERE DISEASE ON DAMAGE TO THE HUMAN COMES FROM TWO THINGS. ONE OF COURSE IS ANEMIA BECAUSE YOU HAVE CONSTANTLY [INDISCERNIBLE] RED BLOOD CELLS. THAT'S PART OF THE CHRONIC ANEMIA BUT THE SEVERE DISEASE IS BECAUSE OF ERYTHROCYTES THAT ARE INFECTED [INDISCERNIBLE] IT'S LIKE THEY HAVE A VELCRO. BECAUSE IF IT DID NOT [INDISCERNIBLE] IT WOULD BE REPLICAS [INDISCERNIBLE] THAT WOULD BE ELIMINATED [INDISCERNIBLE] CAN'T HIDE FROM THESE CLEARANCE MECHANISMS BY THE [INDISCERNIBLE] ENDOTHELIAL CELLS [INDISCERNIBLE] THEN IT CAUSES SEVERE BRAIN MALARIA THAT CAN BE FOUND IN DEATH AND COMAS. AND CHILDREN CAN HAVE ALSO LONG TERM CONSEQUENCES IN TERMS OF BRAIN DEATH. USUALLY BY THE SAME FEMALES HAVE CHILDREN THEY ARE RESENT. YOU HAVE ANTI-BODIES FOR MALARIA IN MANY CASES BUT WHEN THE WOMAN BECOMES PREGNANT, THEN THE PLACENTA CREATES A NEW PLACE WHERE THE PARASITES CAN HIDE. SO PARASITES WITH PARTICULAR FORMS OF THESE ADHERENCE CAN HIDE IN THE PLACENTA AND CAN CAUSE SEVERE MALARIA IN THIS PERSON. AND ALSO THE FINDING OF THE ERYTHROCYTES OF THE PLACENTA DISRUPTS THE FLOW OF BLOOD IN THE PLACENTA AND THE DAMAGES PROCEED IT. THESE HAVE SEVERE CONSEQUENCES. IT'S ESTIMATED EVEN TODAY AFTER INTRODUCTION OF [INDISCERNIBLE] BUT EVERY 45 SECONDS A CHILD DIES OF MALARIA. AND EVERY YEAR 10,000 WOMEN AND 200,000 OF THEIR INDUNLTS DIE AS -- INFANTS DIE OF MALARIA. SO THIS IS SOMETHING THAT IS OF COURSE OBVIOUS IS THAT THE [INDISCERNIBLE] BITE FROM A MOSQUITO. I THINK NOWADAYS EVERYONE ACCEPTS THAT EVERYWHERE IN THE WORLD MALARIA HAS BEEN RERAD INDICATED [INDISCERNIBLE] IF YOU LIVE IN AFRICA YOU WILL GET A 400% COVERAGE MEANING EVERYONE IN THE PLACE THAT GETS INFECTED WITH MALARIA FOUR TIMES EVERY YEAR. WITH THAT RATE OF REINFECTION NO MATTER HOW GOOD THE DRUGS ARE, NO MATTER IT'S JUST NOT POSSIBLE TO CONTROL. AND THIS IS VERY DIFFICULT BECAUSE IT'S EXTREMELY [INDISCERNIBLE]. WE WORK WITH THE MOSQUITO MOSTLY IN [INDISCERNIBLE] BECAUSE IT'S A MAJOR VECTOR IN AFRICA AND THIS MOSQUITO IS REMARKABLY EFFICIENT [INDISCERNIBLE] AND THIS IS ONE OVER REASONS WHY MALARIA IS SUCH A BIG PROBLEM IN AFRICA. MUCH MORE SEVERE THAN OTHER PLACES BECAUSE TRANSMISSION IS MUCH MORE EFFICIENT. SO WHILE I'M GOING TO TALK I'M GOING TO SPEND TIME ON THIS FIRST TWO PARTS BASICALLY GIVING YOU A MODEL WHAT WE HAVE LEARNED [INDISCERNIBLE] UP TO WHAT HAPPENS TO A MOSQUITO WHEN THE PARASITES INVADES THE MID GUT AND TRIES TO CONTAIN THE INFECTION AND HOW THE RESPONSES ALSO AFFECT THE RESPONSE OF THE COMPLEMENT SYSTEM SO THE MOW -- MOSQUITO HAS A COMPLEMENT LIKE SYSTEM [INDISCERNIBLE] AND IS VERY POWERFUL. IN SOME CASES IT CAN COMPLETELY BLOCK TRANSMISSION. I'M GOING TO TALK A LITTLE BIT ABOUT CENTRALLING PATHWAYS AND HOW THEY CAN REGULATE SOME OF THESE. AND I WOULD LIKEIWl TO SPEND MOST OF MY TIME ON THIS PART AND IT'S SHOWING YOU THE EVIDENCE WE HAVE THAT THE PLASMODIUM FALCIPARUM ENVISION OF THE MOSQUITO FAILS AND THAT'S WHEN YOU HAVE A REALLY HIGH TRANSMISSION OF MALARIA. SO THE MOSQUITO, OWE I'M GOING TO TALK ABOUT THE [INDISCERNIBLE] BUT IT APPLIES TO MALARIA. SO THE MOSQUITO IS GETTING [INDISCERNIBLE] MATURATION AND [INDISCERNIBLE] IT'S A BANANA-SHAPED PARASITE THAT MOVES. YOU SHOULD SEE THE VIDEOS, IT'S AMAZING. IT'S JUST CRAWLS AND IT'S ABLE TO INVADE THE [INDISCERNIBLE] INSERTS ITSELF THROUGH THE CELL AND CROSSES IRREPARABLE DAMAGE TO THE INVADED CELL. BUT THE CELL DIES BUT PUTTING A FIGHT UNTIL THE END. THE CELL WILL RESPOND TO INVASION BY TRIGGERING POINTS PRODUCTION OF CHEMICALS THAT ARE TOXIC NITROGEN, NITRATION, REACTION. AND [INDISCERNIBLE] RESPONSES AND IT EMERGES FROM THE CELL AND THEN IT HAS A COMPLEMENT SYSTEM FOR THE MOSQUITO. THERE ARE PROTEINS THAT ARE USED IN THE [INDISCERNIBLE] WHICH IS LIKE THE SERUM, THE REGULATORY SYSTEM. AND THESE CAN BIND TO THE SURFACE OF THE PARASITE AND VERY FAMOUS [INDISCERNIBLE] VERY WELL STUDIED IS THE [INDISCERNIBLE] THE SURFACE OF THE PARASITE AND FORMS A COMPLEX [INDISCERNIBLE] ALL THE COMPONENTS ARE [INDISCERNIBLE]. NOW SOMETHING THAT WAS VERY PUZZLING IS THAT YOU CAN INFECT A MOSQUITO WITH A MOUSE PARASITE THAT'S COLONIAL. SO GENETICALLY ALL OF THE PARTS ARE IDENTICAL. WHEN YOU SEE IT TOOK A BITE ON THE SURFACE OF ONE ON OF ON THE PARASITES [INDISCERNIBLE] HOW COME IT BINDS TO SOMEONE THAT IS NOT IDENTICAL. YOU CAN SEE THEY ARE IDENTICAL. SO WHY? AND THE PICTURE THAT IS EMERGING IS THAT ALTHOUGH THE ONE THAT EXECUTES THE LIKENESS OF THE PARASITE THE FACE [INDISCERNIBLE]. SO BASICALLY WHETHER THE PARASITE WILL SURVIVE OR NOT DEPENDS ON HOW WELL IT FORMS THE EPITHELIAL RESPONSE TO THE INVASION. SO I WILL SHOW YOU SOME OF THE DATA. I CANNOT [INDISCERNIBLE] BUT I'LL GIVE YOU SOME FEELING OF THE [INDISCERNIBLE] TO STUDY THESE. SO THESE ARE SOME PICTURES WHERE YOU CAN SEE WHEN THE)F PARASITE INVADES [INDISCERNIBLE] HERE IN GREEN YOU CAN SEE IT HAS STOPPED [INDISCERNIBLE]. THIS IS A SURFACE PARASITE COATING. YOU SEE THE CELLS [INDISCERNIBLE] AND I WANT TO BE EVENTUALLY [INDISCERNIBLE] THE EPITHELIUM COME TOGETHER AND THEY INVADE [INDISCERNIBLE]. THE CELLS ALSO NOT ONLY EXPRESS HIGH LEVELS OF [INDISCERNIBLE] BUT ALSO ANTI-OXIDANTS [INDISCERNIBLE] IN HUMAN MACROPHAGES IT HAS BEEN SHOWN THAT YOU CAN HAVE NITRATION RESPONSE BUT YOU CAN HAVE A SECOND SET [INDISCERNIBLE] POTENTIALLY THE EFFICIENCY OF THESE. SO YOU THINK WHEN YOU HAVE THE HYPOTHESES THAT THESE ARE HAPPENING IN MOSQUITOS, WE IDENTIFY [INDISCERNIBLE] WITH RESPONSES. AND I WILL SHOW YOU SOME OF THE DATA THAT'S PUBLISHED BUT I WOULD JUST SUMMARIZE HERE. YOU SEE THIS ACCUMULATES IN THE [INDISCERNIBLE] INVADED BY THE PARASITES. WE CAN MEASURE NITRATION [INDISCERNIBLE] COMPARE THE LEVEL OF NITRATION IN A CONTROLLED [INDISCERNIBLE] BUT IF YOU HAVE THE [INDISCERNIBLE] YOU REVEAL INDEMNIFY TRAIKS AND YOU GET THE CONTROL THAT WAS INFECTED WITH [INDISCERNIBLE] PRESENT IN THE MOSQUITO. BUT IF YOU INJECT DOUBLE STANDARD [INDISCERNIBLE] REDUCE EXPRESSION, YOU GET NITRATION. FROM THIS NITRATION IN THE [INDISCERNIBLE] YOU HAVE HIGHER [INDISCERNIBLE] THIS IS A GROUP OF MOSQUITOS, 41 MOSQUITOS AND THIS IS A NUMBER OF PARASITE IN EACH MOSQUITO AND YOU CAN SEE THAT THE MEDIAN LEVEL OF INFECTION INCREASES SIGNIFICANTLY WHEN THE EXPRESSION OF THIS. SO THIS IS TO GIVE YOU [INDISCERNIBLE] ALSO WORKS WITH [INDISCERNIBLE] AND WE FOUND SOME GENETIC EVIDENCE THERE WAS A LINK BETWEEN THE CHEMICAL INDEMNIFY TRAITIONZ AND OXIDATION. SO WE TRIED TO FIND SOME DIRECT EVIDENCE AND HERE IS A PICTURE [INDISCERNIBLE] JUST FIB SHRINK INVADING THE MOSQUITO. SOME OF THEM ARE [INDISCERNIBLE] HERE IN GREEN AND SOME ARE NOT. SO THIS IS THE NUMBER OF PARASITES GAINING CONTROL AND THIS IS THE [INDISCERNIBLE] AND YOU CAN [INDISCERNIBLE] IN TWO DIFFERENT WAYS. ONE IS USING THE PERCENT OF [INDISCERNIBLE] THERE'S A NUMBER OF PARASITES IS REVIEWED SIGNIFICANTLY WITH THE [INDISCERNIBLE] AND WE CAN ALSO MEASURE BINDING TO THE PART [INDISCERNIBLE] SO THERE IS A CONNECTION BETWEEN THE INITIAL RESPONSE AND THE COMPLEMENT. I WOULD LIKE TO TAKE A MINUTE AND I THINK IN IMAGES SO IF IT GETS A LITTLE BIT COMPLICATED BUT I THINK IF WE HAVE SOME IMAGES [INDISCERNIBLE]. LET'S A MOSQUITO THAT JUST HAD A BLOOD MEAL, IT WAS INFECTED SO LET'S LOOK INSIDE. THIS IS THE PARASITE [INDISCERNIBLE] CROSS THE BARRIERS THE BLOOD MEAL THROUGH THE EPITHELIUM [INDISCERNIBLE] INDIRECT CONTACT WITH THE EPITHELIAL. THIS IS MICRO BIOTIC CONTACT WITH THE CELLS [INDISCERNIBLE] LET'S SEE HOW THESE [INDISCERNIBLE]. AS I SHOWED YOU [INDISCERNIBLE] IT'S VERY REACTIVE BITE BECAUSE IT'S SO REACTIVE IT DECAYS QUICKLY INTO NITRITES. IF YOU WANT A STRONG REACTION YOU HAVE TO ACCUMULATE THE NITRATES AND YOU HAVE A [INDISCERNIBLE] GIVE SOME HYDROGEN PEROXIDE AND HAVING THESE COMPONENTS TOGETHER YOU CAN DO A SECOND SET WITH [INDISCERNIBLE] WHICH IS A SECOND CHEMICAL THAT IS VERY REACTIVE. NOW [INDISCERNIBLE] MODIFIED, IT'S CHANGED SOMEHOW. IT DOESN'T KNOW WHAT IS THE NATURE OF THAT MODIFICATION. SO IF IT HAS [INDISCERNIBLE]. SO THIS IS A CASE WHERE THE NEW SYSTEM [INDISCERNIBLE] BUT IF YOU HAVE INVASION AND YOU HAVE THE INDUCTION [INDISCERNIBLE] AND YOU HAVE NITRATES. BUT IF YOU CAN BLOCK THE [INDISCERNIBLE] WE CAN DO IT BY SILENCING EXPRESSION [INDISCERNIBLE] WILL COME OUT BUT THE [INDISCERNIBLE] DIDN'T HAPPEN. THE NITRATION IS NOT AFFECTED SO THESE PARTS ARE THEN [INDISCERNIBLE]. SO THESE PARASITES THAT IS NOT VISIBLE THEN CAN GO AHEAD, DEVELOP, MULTIPLY,AVRELY THOUSANDS [INDISCERNIBLE] MOSQUITO BITES . SO LET'S TALK A LITTLE BIT ABOUT [INDISCERNIBLE] AND THIS IS GOING TO BE VERY BRIEF. I JUST WANT TO MENTION THAT WE HAVE EVIDENCE THAT THE PATHWAYS HAVE A CRITICAL ROLE AND SO INTRACELLULAR AND IN HUMANS [INDISCERNIBLE] MULTIPLE RESPONSES IN THE IMMUNE SYSTEM [INDISCERNIBLE] IT'S INVOLVING THE DEVELOPMENT [INDISCERNIBLE] RESPONSES. I'M SHOWING HERE SOME OF THE PATHWAYS, SHOWING THE GEO MEDICAL [INDISCERNIBLE] GENES AND MOSQUITOS. AND THEN IF YOU FIND EXPRESSION OF THE GENES REQUIRED TO [INDISCERNIBLE] PARASITE CANNOT BE TURNED ON [INDISCERNIBLE]. YOU CAN ALSO FIND THESE SUPRESSORS WHICH NORMALLY% IF YOU BREAK THE SYSTEM IT TURNS OFF [INDISCERNIBLE] OVERACTIVATE THE PATHWAY. SO YOU CHECK A CAR WHEN YOU REMOVE THE BRAKES OR IT'S GOING TO BE OVERACTIVATED. SO WHAT HAPPENED. THESE ARE SAMPLES [INDISCERNIBLE] ACTIVATE THE PATHWAY. IF YOU [INDISCERNIBLE] THE PARASITES DO BETTER. IF YOU [INDISCERNIBLE] IF YOU OVER ACTIVATE THE PATHWAYS YOU HAVE THE [INDISCERNIBLE] NOW THE PARASITES DO WORSE THAN [INDISCERNIBLE]. SO WE ALSO IN THE STUDY TRYING TO UNDERSTAND HOW THESE [INDISCERNIBLE] WAS WORKING AND TO MAKE A LONG STORY SHORT WE FOUND THAT THE PATHWAY [INDISCERNIBLE] AND THESE ARE REQUIRED TO HAVE GOOD NITRATION. IF IT'S NOT INDUCED THEN INDEMNIFY TRAIKS ISN'T EFFECTIVE AND THEN YOU GET [INDISCERNIBLE]. I WOULD LIKE TO TAKE A MINUTE HERE AND JUST SUMMARIZE WHAT I'VE SAID SO FAR BECAUSE IT'S GOING TO BE A CRITICAL FOR THE NEXT PART. SO I WANT TO INVITE YOU TO IMAGINE THAT YOU'RE SITTING [INDISCERNIBLE], RIGHT. A GIANT [INDISCERNIBLE] IS GOING TO WALK THROUGH THE DOOR, GIANT IS GOING TO ENTER. AN ALARM IS SET OFF, RIGHT. THIS ALARM WILL TELL THE SYSTEM THIS IS A FORBIDDEN ZONE. THIS IS A BARRIER ZONE. NOTHING SHOULD CROSS THIS. NOW WHEN THIS HAPPENS THEN THE ALARM IS SET OFF AND A CHEMICAL IS RELEASED INSIDE THE ROOM. THIS CHEMICAL IS NOT STRONG ENOUGH TO KILL THE PARASITE BUT WE CAN THINK OF IT LIKE A FLUORESCENT PAINT. THE KINASE IS PASSING THROUGH THESE CELLS AND A FLUORESCENT PAINT WILL BE MADE THAT WILL HAVE [INDISCERNIBLE] AND GETS OUT, IT'S COVERED IN FLUORESCENT PAINT. SO IT WILL BE IMMEDIATELY RECOGNIZED BY THE SECURITY SYSTEM. NOW IF THE PARASITE MANAGES TO CROSS THE ROOM WITHOUT BEING LABELED, SO THAT FOR INSTANCE [INDISCERNIBLE] THERE'S NO ALARM, THREE NO DIRECTION OF PAINT, IT CAN GO THROUGH, GETS OUT AND THE SECURITY SYSTEM DOESN'T DETECT IT. THAT MEAN THE PARASITE IS NOT THERE. DOES ANYONE HAVE QUESTIONS SO FAR UP TO HERE? OKAY. SO NOW WE'RE GOING TO SEE WHAT HAPPENS. SO WHEN, TO GIVE YOU AN IDEA HOW STRONG THE SYSTEM CAN BE, INFECTION IN A NORMAL MOSQUITO. YOU HAVE BY FA THE NUMBER OF PARASITES. THIS KILLS 3W-8 0% IN THIS. PEOPLE WERE ALL EXCITED ABOUT STEP ONE AND THEN PEOPLE START TRYING [INDISCERNIBLE] AND THIS IS WHAT HAPPENED. [INDISCERNIBLE] LINE OF AFRICAN ORIGIN WITH SOME AFRICAN MOSQUITO. YOU FIND STEP ONE AND NOTHING HAPPENS AND THAT SHOCKED OUR COMMUNITY. ONE INTERPRETATION IS WE HAVE LOST 15 YEARS OF OUR LIVES BARKING UP THE WRONG TREE AND ALL THEY MODELS ARE IRRELEVANT TO HUMANS. BUT THAT IS NOT THE CASE LUCKILY. WHAT I HOPE TO CONVINCE YOU IS THAT WE NEED TO SEE THESE TO KNOW HOW THE SYSTEM CAN WORK. NOW THE QUESTION IS WHY IS IT NOT WORKING HERE. IS THIS NORMAL OR IS THERE SOMETHING UNUSUAL ABOUT THIS. WE STARTED TO LOOK AT THIS MORE DEEPER AND I WILL SHOW YOU THE EVIDENCE WE HAVE THAT PLASMODIUM FALCIPARUM [INDISCERNIBLE] MOSQUITO. SO THIS STARTED THAT WAS CONNECTED WITH NIH ABOUT 20, 25 YEARS AGO. AND THIS IS THE RESULT. IN THIS LINE THE PARASITES DEVELOPED THE [INDISCERNIBLE] BUT AS SOON AS THEY COME IN CONTACT WITH A [INDISCERNIBLE] IN THE STEP ONE, THEN THE PARASITE ARE DAMAGED. BUT IN THIS CASE WHEN THEY ARE KILLED THEY ARE IMMEDIATELY COVERED WITH A BLACK PIGMENT. AND SO COVER THIS BLACK PIGMENT CALLED MELANIN SO THE [INDISCERNIBLE] AND THIS IS NICE BECAUSE WE CAN SEE WHEN THE PARASITE HAS BEEN [INDISCERNIBLE] AND THIS IS VERY USEFUL TO US. SO WHEN WE [INDISCERNIBLE] WAS INITIALLY SELECTED TO KILL ALL PLASMODIUM FROM THE [INDISCERNIBLE] BUT THEN THEY STARTED TESTING OTHER PARTS ONCE THE [INDISCERNIBLE] AND ALL OF THE [INDISCERNIBLE] YOU CAN IMAGINE WITH ONE EXCEPTION. IF YOU INSPECT THIS LINE, IT'S PLASMODIUM FROM AFRICA FROM WHERE THESE MOSQUITO COMES FROM. THEN IT GOES REALLY WELL. SO THAT BEGINS TO SAY THERE'S SOMETHING ABOUT AS FAR AS THAT STRAIN COMES FROM. AND WHETHER MOSQUITO WILL SEE IT OR NOT SEE IT. SO TO GIVE YOU AN IDEA OF HOW DRASTIC SOME OF THIS IS, THIS IS AN AFRICAN [INDISCERNIBLE] AND IN THIS EXPERIENCE I'M GOING TO BE SHOWING YOU THE NUMBER OF PARASITES IN THE MOSQUITO AND THEN I HAVE THE [INDISCERNIBLE] Y AXIS IS THE NUMBER OF LIVE. IN THIS ONE IN THE AFRICAN [INDISCERNIBLE] ALL OF THEM WERE ALIVE. WHILE IF YOU START WITH THE PRECEDING STRAIN THEN NOW BASICALLY ALMOST EVERYBODY 97% ARE DEAD AND BASICALLY NO ONE IS ALIVE. SO THE WAY THESE MOSQUITOS RESPOND AND THIS IS MOST OF THE [INDISCERNIBLE] BUT IT DEPENDS WHERE THE PATIENT INFECTION CAME FROM. IT WAS SOMETHING DIFFERENT. NOW BECAUSE BEFORE FALCIPARUM THEN YOU CAN CROSS THESE, YOU CAN DO A GENETIC CROSS. AND WE, THESE TWO LINES HAVE ALREADY BEEN GONE AT NIH H TO MAP [INDISCERNIBLE]. AND WE WANTED TO KNOW BEFORE WE [INDISCERNIBLE] WE DECIDED TO KNOW WHY ARE THESE BIND. THEY DIE. AND IF YOU INFECT THE MOSQUITO [INDISCERNIBLE] THEY ALSO DIE. BUT IF YOU [INDISCERNIBLE] SO THAT THE MOSQUITO IN THE SYSTEM [INDISCERNIBLE] VERY WELL, 9 % OF THEM. SO THE PARASITE IS KILLED BECAUSE THE MOSQUITO SEES SOME ACTIVITY [INDISCERNIBLE]. NOW WHEN YOU HAVE STEP ONE IN THE AFRICAN STRAIN, WHAT YOU SEE IS THE MEDIAN IS THE [INDISCERNIBLE]. A NUMBER OF [INDISCERNIBLE] DECREASES FROM 4% TO NOTHING BUT OTHER THAN THAT, YOU SEE THAT 9 % OF THE PARASITES ALREADY CAN ESCAPE THE IMMUNE SYSTEM. SO THIS IS A [INDISCERNIBLE] RESPONSE, THIS IS VERY EFFECTIVE IMMUNE RESPONSE. WHY IS THE MOSQUITO RESPONDING SO DIFFERENT IN THESE TWO PARASITES. SO WE DID A COMBINATION OF [INDISCERNIBLE] FROM A GENETIC CROSS BETWEEN THIS AFRICAN AND THE BRA JILLIAN -- BRAZILIAN STRAIN AND WITH THE [INDISCERNIBLE], SORRY, THE [INDISCERNIBLE]. AND THEN YOU SEE A COMBINATION OF THESE TWO WE WERE ABLE TO MAP A [INDISCERNIBLE] THEN WE WERE ABLE TO DETERMINE THAT IT IS [INDISCERNIBLE] ONES THAT ALLOWS [INDISCERNIBLE] AND I WILL SHOW YOU THE DATA FOR THAT IN A MINUTE. THIS IS VERY INTERESTING WAS THE [INDISCERNIBLE] IS ANOTHER GENE CALLED 48-45. THIS IS THE GENE EXPRESSING [INDISCERNIBLE]. ALSO VERY CLOSE TO HERE THERE'S SOMETHING CALLED [INDISCERNIBLE] THE MARKERS THATP)E HIGHLY ASSOCIATED WITH [INDISCERNIBLE]. SO [INDISCERNIBLE] AT THE END WHEN I SEE WHAT IS THE RELEVANCE OF THESE. OKAY. SO IF THE PARASITE HAS THE [INDISCERNIBLE] AS I TOLD YOU ARE MOST OF THEM DO WELL. IN THIS CASE [INDISCERNIBLE] BUT IF YOU REMOVE THESE GENES, SO YOU HAVE A 47 KNOCKOUT. NOW THE AFRICAN [INDISCERNIBLE] AND DIE. SO IF YOU REMOVE THESE GENES [INDISCERNIBLE] 47 FROM THE AFRICAN PART, IT NO LONGER PROVIDES IN THIS CELL. IF YOU REMOVE THE GENE IN THE [INDISCERNIBLE] NOW IT'S DYING, BUT IF YOU INFECT THE MOSQUITO [INDISCERNIBLE] BUT IF YOU [INDISCERNIBLE] WILL SURVIVE EVEN IF THEY DON'T HAVE THE [INDISCERNIBLE]. THE GENE IS ONLY REQUIRED TO AVOID THE MOSQUITO. IT DOESN'T NEED A GENE [INDISCERNIBLE] BUT IT'S JUST TO BE ABLE TO [INDISCERNIBLE] THE MOSQUITO. SO, THIS IS THE [INDISCERNIBLE] ALSO NECESSARY THIS IS THE WILD TYPE. THIS IS A MOW SKEET THAT IS [INDISCERNIBLE] THE PARASITES. BUT IF YOU [INDISCERNIBLE] THIS IS ANOTHER STRAIN. THIS IS FROM THE [INDISCERNIBLE] STRAIN. WHEN YOU DON'T HAVE [INDISCERNIBLE] 47, THEN YOU HAVE VERY LOW INFECTION. SO EVEN IN STRAINS THAT ARE [INDISCERNIBLE] MOSQUITOS KILL THE PARASITES AND [INDISCERNIBLE] DIFFERENT GENETIC BACKGROUND OF THE MOSQUITO BUT YOU ALSO [INDISCERNIBLE] GET VERY LITTLE INFECTION. BUT IF YOU FOUND YOUR COMPLEMENT NOW YOU CAN GET HIGH LEVELS OF INFECTION. SO, I DIDN'T MENTION IT HERE BUT WE WERE ALSO, WE ALSO SHOW AT THAT TIME IF YOU TAKE THE KNOCKOUT LINE AND YOU COMPLEMENT WITH THE AFRICAN OR THE [INDISCERNIBLE] PHENOTYPE BUT IF YOU COMPLEMENT WITH THE BRAZILIAN ALLELE YOU DO NOT [INDISCERNIBLE]. SO ALSO WANTED TO KNOW IF [INDISCERNIBLE] 47 WAS EXPRESSED ON THE SURFACE [INDISCERNIBLE] IT WAS IMPORTANT TO SHOW WHETHER IT WAS [INDISCERNIBLE] IT'S ALSO IN THE PRESENCE OF WILD TYPE [INDISCERNIBLE] HERE BUT IT'S NOT [INDISCERNIBLE] ON THE KNOCKOUT. WHEN WE LOOK AT [INDISCERNIBLE] RESPONSE TO A WILD TYPE VERSUS A KNOCKOUT IS THERE A DIFFERENCE [INDISCERNIBLE]. WHAT WE FIND IS THAT THE W50EU8D TYPE THAT HAS PFS47 HAS LITTLE NITRATION. THE LEGAL OF NITRATION IS LOW THAN THOSE SITTING ON UNAFFECTED BULLET. THE OPPOSITE IS FOR THE KNOCKOUT AND YOU HAVE THE KNOCKOUT THE INDEMNIFY TRAIKS IS MUCH HIGHER [INDISCERNIBLE]. ALSO WE FOUND THAT WE HAVE PSF47 NO LONGER INDUCED EXPRESSION [INDISCERNIBLE] SO OUR CURRENT WORKING IS THE PLACEBO [INDISCERNIBLE] AND WE'RE DOING A SERIES OF EXPERIMENTS. BASICALLY THE [INDISCERNIBLE] IT INHIBITS INDUCTION OF THE POTENTIATION AND IT HAS THE [INDISCERNIBLE] THEN IT WILL BE MODIFIED BY OBSERVATION OF [INDISCERNIBLE] AND THEN THE SECOND SYSTEM WILL BIND [INDISCERNIBLE] AND THEN THESE PARTS WILL DIE AND NOT GO ON. NOW, THERE'S SOMETHING VERY INTERESTING ABOUT PFS47 SO WE'LL GO FROM THE LAST TO THE REAL WORLD. WE'RE GOING TO MAKE A BIG JUMP. SO IF THESE ARE JUST BAR CODES JUST TO LOOK AT DIFFERENT [INDISCERNIBLE] OF PFS47, 48/45 WITH CONNECTIONS WE HAVE AT NIH H. EACH DIFFERENT BAR CODE HAS A DIFFERENT COLOR TO ZIMA LAYER YEAH ORIGINATING IN AFRICA SO YOU GET MORE DIVERSITY IN AFRICA. BUT YOU SEE SOMETHING IN THESE SAMPLE SIZE. THERE'S ONE [INDISCERNIBLE] AND IT'S VERY COMMON AND AMERICA'S THE ONLY ONE WE FIND, ASIA IS THE DOMINANT ONE AND THERE ARE [INDISCERNIBLE] IN AFRICA THAT YOU DON'T SEE OUTSIDE OF THAT. THIS IS ALREADY [INDISCERNIBLE] THIS IS A SMALL [INDISCERNIBLE] IN THIS CASE THREE [INDISCERNIBLE] AND WHEN YOU GO OUT OF AFRICA YOU FIND TWO OF THEM IN ASIA AND THEN TWO DIFFERENT ONES IN THE NEW WORLD. NOW IT HAS BEEN HARD TO EXPLAIN WHY DO YOU GET GEOGRAPHIC FRACTURING. WHAT WE WOULD LIKE TO PROPOSE IS THAT IS THE BEST [INDISCERNIBLE] BY MOSQUITO. IT TAKES A MINUTE TO LOOK AT A LITTLE BIT OF THE HISTORY OF A [INDISCERNIBLE] SO JUST TO SIMPLIFY THESE, THIS IS A WORLD MAP AND THESE ARE THE SPECIES OF MALARIA IN DIFFERENT PLACES. SO JUST AN OVERVIEW I WANTED TO REALIZE THERE ARE MANY DIFFERENT COLORS DIFFERENT REGIONS. MOSQUITOS ARE LOCAL. THE SECOND THING IS THAT THE MALARIA ORIGINATING IN AFRICA [INDISCERNIBLE] BUT THESE MOSQUITOS GO [INDISCERNIBLE] SEPARATE FROM EACH OTHER A LONG TIME AGO. SO THE [INDISCERNIBLE] UNDERGROUND HIDING FROM THE DINOSAURS. THEY WERE NOT EXTINCT YET. IT'S BEEN A KNOWLEDGE LO LONG TIME THESE MOSQUITOS FROM SOUTH AMERICA AND FROM AFRICA HAVE SEEN EACH OTHER. SO NOW SUDDENLY HUMANS BEGIN TO WALK OUT OF AFRICA AND THEY BEGIN TO MOVE. SO IF A [INDISCERNIBLE] WANTS TO BE TRANSMITTED IT HAS TO BE A NEW VECTOR A LOCAL VECTOR. THOSE PARASITES THAT CAN ADAPT ARE TRANSMITTED AND THE ONES THAT CAN'T MAKE THE JUMP THERE'S A DEAD END AND THE INFECTION WILL NOT BE SPREAD IN THE NEW PLACE. SO NOW WE HAVE ANALYZED IN MORE DETAIL [INDISCERNIBLE] PFS IN 47 IN HUMAN SAMPLES. THIS IS 18 ISOLATES AND WE FOUND 30 DIFFERENT [INDISCERNIBLE] AND YOU SEE THERE'S MUCH MORE DIVERSITY IN AFRICA BECAUSE THIS IS THE ORIGIN. YOU SEE THE ALLELES ABUNDANT IN ASIA ARE DIFFERENT FROM THE ONES IN AFRICA. THE MOST EXPENSIVE ARE THE AMERICAS. SO FAR WE HAVEN'T A SINGLE ALLELE, AN ALLELE THAT IS RARE IN AFRICA. SO HOW DO WE EXPLAIN THIS. WHAT HAPPENS IS THAT MOSQUITOS HERE IN AFRICA VERY WELL ADAPTED TO THEIR VECTORS AND SUDDENLY HISTORICALLY THE NEW WORLD IS DISCOVERED AND WITH THE SLAVE TRADE IT'S ESTIMATED THAT MORE THAN MILLION AFRICANS OF THEM OR MOST OF THEM [INDISCERNIBLE] WERE BROUGHT UP INTO THE NEW WORLD. AND THIS HAPPENED [INDISCERNIBLE] SO NOW THESE PARASITES FOUND THEMSELVES IN HUMANS IN PLACES WHERE THE MOSQUITOS ARE LIKE FROM ANOTHER PLANET, RIGHT. THEY'RE REALLY REALLY DIFFERENT FROM THE ONES WE ARE USED TO. SO WE THINK THAT ONLY VERY FEW ACTUALLY ONLY ONE WE CAN IDENTIFY [INDISCERNIBLE] AND IT'S ONLY THESE PARASITES THE ONES THAT HAVE TRANSMITTED IN THE AMERICAS. SO THIS IS OF COURSE A HYPOTHESES BUT TRYING TO FIND EXPERIMENTAL EVIDENCE TO BEGIN TO SUPPORT THIS HYPOTHESES. I JUST WANT TO CALL YOUR ATTENTION TO [INDISCERNIBLE] YELLOW, THIS IS A VECTOR IN THE CARIBBEAN, CENTRAL AMERICA AND A LITTLE BIT IN COLUMBIA. IN THE COAST IN THE ATLANTIC COAST. SO IF YOU LOOK AT A [INDISCERNIBLE] THE NEW WORLD VECTOR, RIGHT. INFECTION IS VERY LOW. THIS IS HERE AS A [INDISCERNIBLE] A STRAIN THAT'S TO BE PERMISSIVE [INDISCERNIBLE]. SO IF YOU TAKE ANOTHER STRAIN FROM SENEGAL THE SAME STORY. BUT IF YOU TAKE A STRAIN FROM BRAZIL THEN IT'S [INDISCERNIBLE] NOT BEING INFECTED. THAT IS JUST INCOMPATIBLE MAYBE THEY'VE HAD DIFFERENCE IN BIOLOGY OR REALLY IN THE NEW SYSTEM. ABOUT YOU WE HAVE BEENŤ— ABLE TO IDENTIFY [INDISCERNIBLE] DECOMPONENT OF THE COMPLEMENT IN THIS SPECIES [INDISCERNIBLE] YOU CAN SEE THE AFRICAN PARASITE [INDISCERNIBLE] NOW YOU GET REALLY GOOD TRANSITION INFECTION [INDISCERNIBLE]. SO WHAT THIS IS TELLING US IS THIS IS GIVING EVIDENCE THAT IT IS THE MOSQUITO IMMUNE SYSTEM ONE THAT IS LIMITING TRANSMISSION FROM [INDISCERNIBLE] SO AN AFRICAN WAS NEVER ABLE TO MAKE A JUMP INTO THE NEW WORLD BECAUSE THE MOSQUITO IS VERY ABLE TO [INDISCERNIBLE]. SO RIGHT NOW WE ARE CONSTRUCTING WHERE WE ARE MAKING THE PSF47 [INDISCERNIBLE] WE'RE TAKING ONE PARASITE AND WE'RE CHANGING THE ALLELE FROM AFRICA FROM THE AMERICAS AND NOW WE HAVE VECTORS IN THE COLLECTION AT NIH H THAT CONQUERS MORE REGIONS OF THE WORLD TRYING TO SEE WHAT IS THE COMPATIBILITY. AND WE THINK THAT THERE ARE MATCHES AND DISMATCHES. IN THIS WASTE WE FEEL THIS IS THE MOST EXTREME SITUATION WHERE WE THINK IT'S A YES OR NO ANSWER [INDISCERNIBLE] SAFER GRADE HERE. AND THIS IS IMPORTANT BECAUSE IF WE UNDERSTAND WHAT IS THE KNOWLEDGE OF COMPATIBILITY WE CAN PREDICT WHAT IS THE CHANCE THAT A [INDISCERNIBLE] FOR EXAMPLE WE HAVE I TALK ABOUT THE [INDISCERNIBLE] THAT IS ASSOCIATED WITH THE [INDISCERNIBLE] CAMBODIA. IF YOU HAVE THIS RESISTANCE, WHAT IS THE CHANCE THAT THESE [INDISCERNIBLE] FOR INSTANCE WERE PROMISING A NEWER WORLD BASED ON THESE BUT [INDISCERNIBLE] NEVER WERE ABLE TO ESTABLISH IN THE NEW WORLD. CAN THEY JUMP BACK TO AFRICA, I THINK YES. SO I THINK AS WE UNDERSTAND THE MOLECULAR BASIS OF THE COMPATIBILITY IT CAN REALLY GIVE YOU AN IMMUNLOGICAL WHICH PARASITES ARE LIKELY TO SPREAD IN AN AREA AND UNDERSTAND WHAT MAKES COMPATIBLE OR NOT. SO OF COURSE MANY QUESTIONS STILL ARISE AND WE'RE TRYING TO UNDERSTAND HOW THIS WORKS. FOR EXAMPLE IF THERE IS THE PFS47 THAT'S [INDISCERNIBLE] THERE HAS TO BE A UNIT RECEPTOR IN THE MOSQUITO WHERE THERE'S A COUNTERPART. WE'RE WORKING HARD TO IDENTIFY AND IT'S ALSO HAVING PFS47 IS THE KEY TO A VERY SUCCESSFUL SPREAD OF MALARIA COULD THAT BE A WEAK POINT IN THE SYSTEM [INDISCERNIBLE] AND NOW WE MAKE THESE PARASITES VISIBLE [INDISCERNIBLE]. SO THIS IS WHERE WE ARE WORKING. OF COURSE OUR LONG TERM GOAL IS TO DISRUPT TRANSMISSION TO PREVENT HUMAN DISEASE AND [INDISCERNIBLE] QUALITY OF LIFE OF THOSE ENDEMIC AREAS. I WANT TO BRIEFLY INTRODUCE SOME OF THE PEOPLE WHO HAVE DONE THIS WORK. [INDISCERNIBLE] HEADING THE PROJECT, WE FOUND A CONNECTION BETWEEN NITRATION [INDISCERNIBLE] DID THE WORK WITH THE [INDISCERNIBLE] IS A SCIENTIST IN THE LAB AND [INDISCERNIBLE] SEVEN YEAR PROJECT [INDISCERNIBLE] VERY CHALLENGING BUT AT THE END I THINK IT WAS WELL FURTHER IT. AND THEN THE PEOPLE FROM THE LAB THAT ARE WORKING VERY HARD TRYING TO UNDERSTAND HOW THIS WORKS. AND ALSO SOME COLLABORATORS [INDISCERNIBLE] WE HAVE INTERVIEWS [INDISCERNIBLE] AND WE'RE ASKING IS IT ENOUGH TO HAVE THE [INDISCERNIBLE] BECOMING INVISIBLE, IS THAT ENOUGH. AND WE SHOULD HAVE THE [INDISCERNIBLE] TOMORROW AFTERNOON SO WE'RE HOPING THAT THAT WILL WORK AS IT WILL BE AMAZING. [INDISCERNIBLE] CRYSTAL STRUCTURE AND TRYING TO UNDERSTAND HOW IT WORKS. AND [INDISCERNIBLE] IN CAMEROON [INDISCERNIBLE]. I WOULD LIKE TO STOP HERE AND THANK YOU. [APPLAUSE] >> THANK YOU. >> WHEN THE [INDISCERNIBLE] >> YES. >> THEY ALL CAME WITH VARIOUS DIFFERENT VERSIONS [INDISCERNIBLE] >> YES. >> WAS THIS JUST LUCK AFTER MANY YEARS [INDISCERNIBLE]. >> THIS IS WHAT WE THINK. THAT THERE WAS ONE AND ACTUALLY A RARE ONE. WE ONLY HAVE ONE SAMPLE IN ALL THE COLLECTION, WE ONLY HAVE ONE. WITH THE NEW WORLD ALLELE THAT CAME FROM AFRICA. BUT THIS IS I THINK WHAT HAPPENED THAT IT WAS ALMOST EQUIVALENT, THERE WAS A HIGH CHANCE IT WOULDN'T HAVE HAPPENED, RIGHT. IT'S JUST THAT THERE WAS ONE PARASITE THAT HAD JUST THE RIGHT COMBINATION. SO WHAT WE'RE DOING IT'S DIFFICULT TO COMPARE BECAUSE WHEN YOU COMPARE [INDISCERNIBLE] CONTINENTS [INDISCERNIBLE] BY HAVING JUST ALL OF THEM [INDISCERNIBLE] AND THE COMPATIBILITY QUESTION WITH EACH OF THE DIFFERENT STRAINS. SO NOW WE HAVE [INDISCERNIBLE] THAT IS AFRICAN PARASITE BUT HAS A NEW WORLD ALLELE EXPRESSING THE RIGHT, EVERYTHING RIGHT AND OUR HOPE IS THIS PARASITE WILL JUMP INTO THE NEW WORLD FOR THE FIRST TIME. SO HAVING THAT JUMP MIGHT BE ENOUGH. BUT SINCE WE HAVE A PRELIMINARY WITH PLASMIDS CONFIRMATION [INDISCERNIBLE] MULTIPLE DRUGS THAT HAVE TO BE [INDISCERNIBLE] WE THINK THE TRANSFORMANCE [INDISCERNIBLE] >> CAROLINA WHAT'S THE EFFECT OF GLOBAL WARMING GOING TO BE ON THE DISTRIBUTION OF MOSQUITOS. >> WELL, I THINK THAT YOU KNOW, I THINK DISTRIBUTION WILL CHANGE. PLACES WHERE WE DID NOT HAVE LET'S SAY COMPATIBLE COMBINATIONS CHANGE [INDISCERNIBLE] TRANSMISSIONS BASICALLY YOU HAVE NOT SEEN THEM BEFORE. I THINK IT'S WORRISOME FOR MOSQUITOS LIKE [INDISCERNIBLE] AND OTHER DISEASES THAT ARE LESS SPECIFIC I THINK [INDISCERNIBLE] IN GENERAL IS SUPPOSED TO BE LOCAL AND I THINK IT MIGHT SPREAD. I THINK THE BIGGEST EXAMPLE OF HOW IMPORTANT THE VECTOR IS IN THE 19 50'S FROM [INDISCERNIBLE] WHICH IS VERY SIMILAR. IT'S A SPECIALS CLOSE TO GANDHI [INDISCERNIBLE] WAS BROUGHT INTO [INDISCERNIBLE] BY MISTAKE. AND THEY BEGAN TO HAVE EPIDEMIC AND VERY HIGH MORTALITY IN THIS AREA. LUCKILY SOMEONE NOTICED AND REALIZED WHAT WAS GOING ON AND THEY DID A [INDISCERNIBLE]. IT DID NOT ACHIEVED THE ERADICATION SOUTH AMERICA WOULD BE LIKE AFRICA TODAY. SO I THINK IT'S THE VECTOR. THE VECTOR IS REALLY CRITICAL. I THINK THE VIRILITY DEPENDS HOW MANY [INDISCERNIBLE] AND THAT'S WHERE IT'S THE MAXIMUM BECAUSE THIS MOSQUITO BASICALLY DOESN'T SEE THE PARASITE SO ALLOWS THE PARASITE TO GO THROUGH. >> WE BRING BACK DDT? >> WELL, I THINK UNDER CERTAIN CONDITIONS SOME PEOPLE THINK THAT IT COULD BE USEFUL. BUT OF COURSE MOSQUITOS DEVELOP VERY QUICKLY SO I THINK [INDISCERNIBLE] LIKE IT WAS GONE BEFORE WITH THE IDEA TO ERADICATE I DON'T THINK SO BUT AS PART OF THE INTEGRATED CAMPAIGNS WITH LONG LASTING [INDISCERNIBLE] TO REDUCE MOSQUITOS WHEN YOU ARE [INDISCERNIBLE]. >> YES? >> I READ A RECENT ARTICLE ABOUT [INDISCERNIBLE] IN BRAZIL AND MODIFIED MOSQUITO I GUESS. DO YOU HAVE ANY OPINIONS ABOUT THAT. >> YES ARE THAT'S A VERY INTERESTING [INDISCERNIBLE] BUT I THINK IT'S A [INDISCERNIBLE] WHAT THEY HAVE IS A [INDISCERNIBLE] THE MOSQUITO MAIL HAS A GENE WHEN THEY MATE THE FEMALES OF THE NEXT GENERATION WILL BE UNABLE TO FLY. SO IT'S A MOSQUITO THAT JUST WILL BE PARALYZED. SO IF YOU RELEASE A LOT OF THESE MALES AND THEY MATE WITH THE LOCAL POP LESION THEY'LL FIND THE FEMALE AND THE MOSQUITO POPULATION WILL COLLAPSE. SO I THINK THIS COULD BE VERY USEFUL FOR INSTANCE IF YOU'RE ABOUT TO HAVE A [INDISCERNIBLE] AND YOU HAVE MOSQUITOS AND YOU HAVE TO DO IT QUICKLY I THINK IT WOULD BE A VERY GOOD MEASUREMENT. THE PROBLEM IS IT'S NOT [INDISCERNIBLE] YOU HAVE TO RELEASE THOUSANDS OF MOSQUITOS EVERY YEAR. SO I THINK IT HAS AN APPLICATION BUT BY ITSELF I THINK IT'S [INDISCERNIBLE] BECAUSE IT'S VERY EXPENSIVE. THERE'S SOME [INDISCERNIBLE] WHICH I DON'T KNOW IF YOU WANT. >> DO YOU THINK THAT THE PFS47 IS AFFECTING, IS CAUSING A SURFACE PROTEIN OR INHIBITING A SURFACE PROTEIN ON THE PARASITE WHICH IS AROUND FOR IT TO BE MARKED LIKE EITHER IT'S INHIBITING? >> THIS IS THE BILLION DOLLAR QUESTION. SO THE SHORT ANSWER IS WE'RE NOT SURE IT'S WORKING BUT WE KNOW WE CAN MAKE RECALL NUNS 47 CAN WHAT WE THINK IS A CRITICAL PART OF [INDISCERNIBLE] THEY SELL IN VITRO DISEASE THE CELL'S NO LONGER ACTIVATED. SOMEHOW THESE CELLS NO LONGER [INDISCERNIBLE] SO IN THIS IMAGE THAT I WAS GIVING YOU IMAGINE THAT SOMEONE DISCONNECT THE ALARM AND THE DOOR [INDISCERNIBLE] BUT IT REALLY SOMEHOW [INDISCERNIBLE] INDUCTION OF [INDISCERNIBLE] THAT PUTTING THESE SIMILARLY CAN BLOCK SIGNALING. NOW THIS IS A [INDISCERNIBLE] WHERE WE ACTUALLY THINK A [INDISCERNIBLE] IN A CELL LINE BUT WE SEE LACK OF INDUCTION OF THE INFECTOR GENE WHEN WE DO THAT. IT'S SIMPLER YOU DON'T HAVE INVASION YOU DON'T HAVE INJURY BECAUSE THIS IS A COMPLEX SYSTEM BECAUSE THE CELL IS UNDER GOING APOPTOSIS. AND SOMETHING [INDISCERNIBLE] IN THIS PICTURE WHICH I THINK IS GOING TO BE CENTRAL PLAYERS, MOSQUITOS HAVE [INDISCERNIBLE] BUT THEY ARE CENTRAL IN THESE. I DID NOT EVEN BRING THEM INTO THE PICTURE BECAUSE IT GETS [INDISCERNIBLE]. BUT I THINK THESE ARE MAJOR PLAYERS SO I THINK THERE'S INTERACTIONS WITH ALL OF THEM BUT IN THE END IT'S THE OUTCOME [INDISCERNIBLE]. >> ARE THERE EFFORTS TO TRY AND INTERFERE, IF YOU COULD BLOCK THE HEPATIC STAGE YOU'D STOP THE CYCLE. >> YES. >> WE SEEM TO BE LEARNING A LOT IN RECENT YEARS OF THE BASIC CELL BIOLOGY HOW THE PA HAT SITE VAC VEUL EIGHTY. >> THERE ARE THREE VACCINES. ONE IS TRANSMISSION BLOCKING A PERSON THAT IS INFECTED WILL NO LONGER TRANSMIT IF YOU GET MALARIA AND MOSQUITO BITES YOU CANNOT INBEEN IF HE CAN MOSQUITOS. LET'S GOOD AT THE [INDISCERNIBLE] BUT IT'S THE BENEFIT OF THE PERSON WHO RECEIVES THE VACCINE. NOW PEOPLE HAVE [INDISCERNIBLE] VACCINES WHICH ARE STAGES BEFORE THE [INDISCERNIBLE] AND MANY ARE WORKING ON THAT. THE PROBLEM IS YOU HAVE A VERY NARROW WINDOW, RIGHT FROM THE MOSQUITO BITES [INDISCERNIBLE] GOES INTO THE LIVER YOU HAVE A FEW MINUTES TO MAYBE HOURS. SO THE TEAM HAS TO WORK VERY QUICKLY, RIGHT THE ONCE IT GOES INTO THE LIVER THE PARASITES MULTIPLY AND THEN ONCE IT GOES OUT. TO GIVE YOU AN IDEA IN MOSQUITO YOU GET ONE OR TWO SURVIVING BUT WITH HUMANS WEDNESDAY YOU GO TO THE [INDISCERNIBLE]pxÖ WE HAVE 10-11 PARASITES. SO THE NUMBERS ARE SPIRALING. ONCE YOU HAVE THAT MANY PARASITES THEN IT BECOMES VERY DIFFICULT TO CONTROL. SOME PEOPLE THINK MAYBE IT'S OKAY TO ALLOW THE INFECTION TO HAPPEN AND WE SHOULD START PREVENTING MALARIA FIRST TRY TO DEVELOP VACCINES THAT WILL PREVENT MALARIA BECAUSE AS LONG AS YOU GET OUT [INDISCERNIBLE] NOW THERE'S SOME NEW PROMISES INFECTUAL STAGE VACCINE THAT WE APPEAR TO UNDERSTAND BETTER WHAT IS THE RIGHT CONFIRMATION AND THE RIGHT WAY TO [INDISCERNIBLE] HAS SOME VERY UNDERSTANDING RESULTS SO THAT THEY CAN BE VERY GOOD ANTIGENS [INDISCERNIBLE] >> HOW DOES VIVAX DEAL WITH THE AWE LAWFERL SYSTEM. >> THAT'S A GREAT QUESTION. SO WE HAVE JUST STARTED WITH WORK WITH THAT. NOW THAT'S A MUCH MORE DIFFICULT BECAUSE THERE'S NO IN VITRO SYSTEM THERE'S NO WAY TO STUDY VIVAX BUT WHAT WE'RE ABOUT TO START DOING IS IN THE NEW WORLD, WE ONLY FOUND ONE ALLELE. NOW WE'RE TRYING TO PUT THIS, WE'RE THINKING OF PUTTING THE VIVAX PFS47 INTO THE [INDISCERNIBLE] KNOCKOUT TO SEE IF THAT'S ENOUGH TO OPEN THE DOORS. MY GUESS IS VIVAX MAY BE MORE ABLE TO JUMP TO DIFFERENT VECTORS BUT I DON'T KNOW. YOU HAVE SPECULATION [INDISCERNIBLE]. STILL WE HAVE SOME SAMPLE THAT ARE FULL OF SAMPLES THAT ARE SMALL AT THE MOMENT BUT SO FAR ALL THE VIE VAC THAT WE HAVE FECALS AND WE HAVE [INDISCERNIBLE] SAMPLES FROM BRAZIL WITH ONLY ONE ALLELE AND IT'S THE SAME ALLELE THAT'S BEEN REPORTED IN [INDISCERNIBLE]. SO MAYBE IT'S ONLY ONE ALLELE BUT WE DON'T KNOW YET. >> SO THEORIES THAT ARE NOW BECOMING MORE OUT IN THE OPEN ABOUT THE ORIGIN OF VIRUSES. WHAT DO YOU THINK IS THE ORIGIN OF A PLASMODIUM. >> THE ORIGIN. >> ORIGIN. >> I THINK THAT MOSQUITOS MAYBE WILL GET INFECTED. THEY SAY THAT THE MOSQUITOS USED TO [INDISCERNIBLE] FROM INSKS THAT TAKES THEM. NOW WHAT IS THE JUMP. THE THING IS WHAT IS IF YOU ARE THE CHICKEN OR THE EGG, RIGHT. SO WHAT'S FIRST WAS THE MOSQUITO FIRST INFECTED AND THEN IT ADAPTED TO HUMANS [INDISCERNIBLE]. OR DID HUMANS HAVE INFECTIONS AND THEN IT JUMP INTO THE MOSQUITO. IT IS HARD TO IMAGINE BECAUSE YOU NEED BOTH TO HAVE A TRANSMISSION. SO HOW WAS THE CYCLE FIRST. I DON'T KNOW, DO YOU HAVE ANY ... >> [INDISCERNIBLE]. ONE OF THE THEORIES THAT PEOPLE HAVE PROPOSED IS BECAUSE THE PART OF THE MALARIA CYCLE THE LIFE CYCLE THAT HAS SEXUAL STAGES AND MOST COMPLEX TO ONES THAT IS IN THE MOSQUITO PROBABLY THAT STATE WAS FIRST. AND THE PARASITE ALSO HAS A [INDISCERNIBLE] WHICH IS AN ANCIENT [INDISCERNIBLE] ORGANELLE AQUATIC ORGANISM MOST LIKELY AND THE MOSQUITO HAS AN AQUATIC STAGE PROBABLY THE FIRST PLASMODIUM WAS THE PARASITE IN THE AQUATIC STAGE OF LARVAE AND MAYBE LATER ON CAME THE ACCIDENTAL HOST. >> MOSTLY PROBABLY THE [INDISCERNIBLE]. THEN SOMEONE BIT A HUMAN AND THEY GOT A SUPER MEAL. OH WOW, THIS IS GOOD SO THEN THEY MULTIPLY. AND THAT'S HOW IT STARTED THAT. >> ARE THERE ANY OTHER QUESTIONS? JUST TWO THINGS. ONE, I'M REMINDED, I THINK THAT THERE HAVE BEEN FOUR MALARIAS THAT HAS BEEN THE GREATEST SUBJECT FOR NUMBERS OF NOBEL PRIZES OF CERTAINLY ANY INFECTIOUS DISEASE. I BELIEVE THERE WERE FOUR NOBEL PRIZES GIVEN THREE OF WHICH WERE REALLY APPROPRIATE. ONE OF THEME FEVER THERAPY HAS SORT OF DIED BY THE WAYSIDE BUT YOU CAN SEE THAT FROM THIS INCREDIBLE EXPLOSION OF INFORMATION AND HOW YOU GET YOUR FEELING, I GOT THE FEELING TODAY THAT WE WERE VERY PRIVILEGED TO BE ON THE CREST OF ALL KINDS OF WAVES THAT ARE COMING. AND WE CAN DO NOTHING BUT ADMIRE THE QUALITY OF YOUR WORK AND YOUR PRESENTATION. THANK YOU VERY VERY MUCH.