The Elucidation of Chronic Diseases Occuring in High Incidence in Primitive and Isolated Populations
Gajdusek, Carleton Daniel
This was Carleton Gajdusek’s first lecture in Lindau after winning the Nobel Prize in Physiology or Medicine two years previously. Gajdusek dedicated most of the lecture to the research that led to the Nobel Prize, namely the occurrence of kuru, a fatal neurodegenerative disease that afflicted the Fore people of eastern New Guinea. The characteristic features of the disease are dementia and loss of muscle function. All kuru sufferers died within one year of the first symptoms taking place. Members of the Fore tribe had a ritual of eating their dead relatives as a sign of mourning, and this was demonstrated as the root cause of kuru, particularly since women and children were most likely to fall victim to the disease, and they usually ate the brains of the dead. Missionaries working in New Guinea discouraged the Fore people from this practice and the illness that was considered to be an epidemic gradually came to a halt, although long incubation times caused the disease to persist in the population for decades. “We are dealing with virus-like agents, we are dealing with a new group of microbes”, noted Gajdusek, however, he used the term “virus” or “slow virus” throughout the lecture. The replicating agent was still a mystery to researchers. At that point it was known that the “virus” was bizarrely devoid of nucleic acid, and resistant to ionising radiation, boiling, and storage in formaldehyde. Its presence in the body did not induce an immune response. The similarities of kuru to Creutzfeld-Jakob Disease (CJD) and to two animal diseases; scrapie in sheep and goats and transmissible mink encephalopathy were confirmed, yet the form of transmission of CJD and the animal diseases was also unknown. Only several years later, in 1982, Stanley Prusiner published a paper demonstrating that scrapie is caused by proteinaceous infectious particles, and proposed the use of the term “prions”. These pathogens induce normal proteins to change their structure, which leads to changes in their physicochemical properties. Prions are responsible for CJD and kuru, as well as several neurodegenerative animal diseases. Prusiner was awarded the Nobel Prize in Physiology or Medicine in 1997 “for his discovery of Prions – a new biological principle of infection”. Hanna Kurlanda-Witek-lecture.pdf-lecture.pdf
 Our German hosts and fellow colleagues and students, I'm very grateful to be with you. I'm coming directly from a much warmer climate about which I hope to speak to you a bit at the end of my lecture. And I'm suffering under the influence of your beautiful weather here. The study of an epidemic of new disease, new to Western medicine, and a chronic, progressive, always fatal central nervous system disease in a remote population isolate in people still in a stone age culture in the central highlands of New Guinea, who, hate their own dead, has a right of respect and mourning to their nearest kinsmen, has led us to two major discoveries. The first, already mentioned by the over-generous introduction, was the discovery that chronic, progressive, even at times totally non-inflammatory, and even apparently heredophomial diseases could be caused by slow-acting virus infections, even years and at times several decades after original infection and a long, silent incubation period. The second discovery was that of a new group of microorganisms, which cause human diseases and which possess such unusual physical, chemical, and biochemical, and biological properties that many of our colleagues prefer that we not call them viruses. They are so very different from all other microorganisms that they clearly represent a new type of replicating agent, which we have tentatively decided to call unconventional viruses. The disease in question that led us to these discoveries is called Khuru, a now disappearing plague of the Foray people and of ten other cultural and linguistic groups in the central highlands of eastern New Guinea who intermarry with them. They all inhabit a high mountainous interior of the eastern highlands, living at from 2,000 to 3,000 meters above sea level. By opening the bodies and the skulls of their dead kinsmen, they contaminated the skin and eyes principally of the women who performed the morning ritual and of the young children with them. This led to a fatal brain disease appearing 5, 10, even 20 or 30 years later. It became the most common death among them. In fact, it accounted in the first 10 years of our total surveillance of all the population involved for over 50% of all death and among adult women, 90% of all those who died in the 15 year period of surveillance died from Khuru. We have now discovered that the disease first described in Germany a half century ago and named Kreuzfeld-Yakob type of presenial dementia after the two German neurologists who first described it is a worldwide disease of at least tenfold and probably a hundredfold the incidents that we attributed to it only a few years ago. It is caused by a virus indistinguishable from that of Khuru, but whereas we know the mode of transmission of Khuru from the contamination of the skin and mucous membranes through the ritual rite of cannibalism, we do not know the usual means of transmission of our civilized form of Khuru throughout the rest of the world. We now know that these two unconventional viruses of man are similar to two such viruses of animals, scrapey, tauberkrankheit of Deutsch or Tremblant en français and a disease called transmissible mink encephalopathy of mink. We call the group after the nature of its neuropathological lesion the subacute spongiform virus encephalopathy. Same resistance to an activation by ultraviolet radiation of 2,540 angstroms to 254 nanometers, the wavelength which is the peak absorption wavelength for RNA and DNA has led to the prediction that Khuru and CJD or Kreuzfeldt-Yakob disease virus and scrapey viruses possess no nucleic acid. This heresy has been further promulgated on the discovery that these unconventional viruses also exhibit an enormous resistance to ionizing radiation using either gamma rays from a cobalt 60 source or neutron beam. We're doing this work with Latigé at the Fondation Curie in Paris as well as at Oak Ridge and we have repeated the work of our British colleagues, Hague and Tigger Alpers, and confirm this resistance which is in the order of 100 to 1,000 times that known for any other microbial system. In fact the equivalent target size for a sphere with such unusual resistance to ionizing radiation as specific, a non-specific, though the damage may be, would be calculated to be under 150,000 Dalton's, a molecular weight which is only one-tenth the size of the smallest genome previously known to virology. Finally, intensive study of these viruses for over a decade has failed to reveal any antigenicity and in both the natural diseases and the experimentally induced diseases in laboratory animals, there is no evidence that the victims develop any immunity or that the immune system plays any role whatsoever in the disorder. The humoral and the delayed hypersensitivity types of immune response, B cell and T cell function remain intact and unstimulated. There are no immune complexes formed and no immune complex deposits found on basement glomerular membranes or in the chloride plexus. These thus are the first mammalium microbial pathogens which apparently contain no non-host protein in their structure. Instead, they seem to be minute messenger RNA sized pieces of DNA tightly bound to host membrane systems. And indeed, the demonstration that they are in fact DNA is very tenuous based on only one unconfirmed very recent experiment now in press. The only structure we are left possible for them at present remains a tightly membrane bound particle of size no larger than 150,000 Dalton's. Being probably for none of its own enzymes or any core or coat proteins. A virus without core or coat leaves those of us in structural virology most bewildered. It is from recent work in plant virology that we found find sucre and the hope that we may remain true to our basic tenets of our current DNA RNA religion and not have to retreat into the previous speculations of replicating membranes devoid of nucleotide structures or replicating basic proteins. But I must warn you that the very same workers who with no difficulty whatsoever quickly established the full sequence of FIX174 the smallest virus of bacteria yet sequenced have been six years on this problem from Caltech and North Carolina out and in Germany and in England and have failed yet to establish unequivocally that a nucleic acid is involved in this whole system. That we are dealing with viruses and viruses that cause human disease is very clear indeed. They grow in vitro in tissue cultures of infected cells obtained by primary culture of the spleen, liver, lymph nodes or brain of infected animals or primary human or animals with the natural diseases. Other cell lines such as L cells have been infected in vitro with the need of causing fusion to get the infection started and the perhaps hazardous procedures of transforming both human and animal cell lines with the SV40 genome so that they bear the tumor antigen and still carry replicating simultaneously the infected genome of both the Croix-Velliacob and the Coru agent and different cell lines and now the Scrapey have all been successfully performed. We are dealing with virus like agents. We are dealing with a new group of microbes which might better be named by another term and as usual the more fortunate plant virologists have preceded us and found that they must also revise their basic microbial conceptions in a group of agents which are the smallest of the plant viruses. They prefer to call following dener the man who has defined them most thoroughly, viroids. I will talk about them later and with that introduction I will start on a series of slides. If I get through them on time I will then try to take you on a historical travelogue to where this current day molecular biology began. So if I pay somewhat short shrift to the rather lavish and hardly credible claims I have made by beguipologies from my molecular biological colleagues to make it a little easier to follow the terminology of a discipline of neurology and neuropathology notorious for the use of impossible eponyms and impossible and almost meaningless descriptions based on neuropathological lesion. I will leave you rest assured that we have introduced only one new name into medicine the name Kuru which is a nice non-anglo-saxon four letter word which has even crept into the Webster Collegiate Dictionary whereas Lewis has yet failed. I am unable to use any other term for Kreuzfeld-Yakob disease than the long established eponym. The disease does run by 17 additional all longer synonyms in the neurological literature. Scrape is a spontaneous disease of sheep and goats throughout the world and minkensophilopathy we now have good reason both virologically and epidemiologically to believe is simply the scrapy infection having spread to mink ranches naturally. In fact it is no one that where it was first seen the carcasses of scrapy infected sheep have been fed. The next slide please. This slide simply shows the natural diseases on the left the experimental diseases in experimentally infected animals after over a year of incubation on the right. It shows what is called status spongiosis or spongiform change by the neuropathologist which simply means a sponge like punched out hole appearance of the gray matter it is not referring to white matter spongiosis and although it was previously thought to be edema and interstellular fluid accumulation it is exactly not that. It is a swelling of neuronal and astroglyos cells based on a coalescence of minute vacuoles which can be seen in their origin by electron microscopy long before they are visible to light microscopists. This is scrapy in sheep and scrapy in mice, curu in a child and curu in a chimpanzee inoculated with the child's brain tissue. Kreuzfeld Jacob from a brain biopsy in Oxford and the same process in the brain two years later of a chimpanzee inoculated with it. Next slide please. The same distribution, scrapy, natural and experimental, curu natural and experimental, Kreuzfeld natural and experimental demonstrating with a specific stain that shows astroglyos cells the fibrous astroglyosis which is a dominant part of the pathological picture and about as severe in this disease and this group of diseases as in any disease we know. On the other hand these are the only infections of the brain and I make no exceptions in infections. I mainly range through the whole range of such things as the protozoa through the viruses in which there is no perivascular cuffing, no leukocytic invasion of the brain parenchyma and no sign of a hypersensitivity or a inflammatory response. Next slide. For those clinically minded, next slide please. The phenomenon that these diseases proceed from beginning to fatal outcome with no change in spinal fluid protein and no pleocytosis at any stage of the disease is a further unique finding in fatal CNS infections. Now I'm not going to dwell on the hundreds of studies attempting to purify scrapy first using cesium chloride then using calcium and potassium tartrate, metrismid as well as sucrose and sucrose saline gradients have been used and the workers who have done this work are not workers who have difficulty using zonal ultrace centrifuges and purifying reverse transcriptases, viral subunits, P30 proteins, polyacrylamide gel, electrophoresing them but these same workers have been unable to purify by these techniques or any other electrophoretic focusing and the like the scrapy agent because no matter what small portion of maximum infectivity at a density of 1.2 to 1.25 they take, it spreads again and electron microscopic control of the re-zonal banding of it simply shows that we are trying to purify soap bubbles. The material obtained with enormously high infectivity is a three-layered bioelectron microscope plasma membrane light structure which fasciculates and the problem is that of concentrating and purifying membranes where the reactive receptor or the reactive macromolecule on the membrane is not known and although we assume it is a small nucleic acid all attempts with hot formaldehyde and with detergents to release this presumed genome have failed to obtain any infectious nucleic acid. Next slide. The most disturbing early data which came first from Compton working on scrapy then we reconfirmed it in using zonal banded and fluorocarbon cleaned brain suspended scrapy virus. We obtained linear inactivation curves at ergs 10 to the per square millimeter times 10 to the fourth. A energy input which if you are not familiar with radiation work would not lead you to realize that the most radio resistant forms of life we know to 254 nanometer radiation would intersect here at six. They would be the U1 variant of tobacco mosaic and bacillus radiodurans. This was interpreted first as absolute proof that the agents could not contain nucleic acid. In the suspensions if the argument is that they are dirty not sufficiently cleaned one puts a large variety of mammalian plant viruses and bacteriophage. These so called dirty suspensions have no effect on protection at this radiation. Next slide. I am not going to dwell on the long discussion this might entail but here is the same data you just saw but it simply shows that the plant extracts of the plant viroids these newly described smallest known viruses are even more radio resistant and the purified RNA genome of 115,000 molecular weight of potato spindle tuber virus is of the same order of magnitude of radio resistance whereas all conventional viruses fall here. Next slide please. This alone eliminates all other plant and animal viruses from consideration. Studies of very unsatisfactory zonal banding into 55 fractions in a sucrose saline gradient that the best anyone has obtained is a log scale so it doesn't plot out as well as the earlier ones I showed you. I have failed to reveal any relation with marker enzyme systems in enzyme activity for three lysosomal membrane bound enzymes and one mitochondrial enzyme. Next slide please. I am now going to rapidly summarize probably as fast as you can read. Much of what I have said simply to drive home the fact that these surely are an unconventional type of viral agent. On the physical chemical side I fail to mention what alone would leave any earlier microbiologist totally skeptical of the integrity of the workers. That a virus or any microbe can be stored in 4% formaldehyde or 10 or 20% formaldehyde at room temperature for years without losing tighter namely embalming fluid and that which we use in pathological laboratories to store specimens is blatantly ridiculous. It happens to be the fact. We have isolated Creutz-Feliaco virus from material that has been stored years at room temperature in pathological laboratories in formaldehyde. Scrapey and minkencephalopathy have been taken off paraffin embedded blocks which were for two years in formaldehyde before they were sectioned and left several years as blocks and obtained as viable microbes. Beta-propreolactone doesn't touch them. Ethylene, diamine, tetraacetic acid doesn't touch them. Proteases and the nucleases on even the deprotonized fluorocarbon treated zonal band infractions are not effective nor can we use hot formaldehyde to inactivate them and searching for double or single stranded RNA. Heat resistance is not incompatible with the biological experience but it is incompatible with most virological experience. Up until 80 degrees we have full stability. At 85 degrees we begin to inactivate and as with the hepatitis virus, boiling is not sufficient for hospital sterilization. Autoclaving however most certainly is. Next slide please. To go on further I have already summarized the UV resistance for you. I've mentioned the enormous ionizing radiation resistance. We use ultrasonic energy in such energy input that we would be inactivating rapidly polio or flu or most conventional viruses and all we do is gain in yield increasing our infectivity tighter. When we do an action spectrum at 2,370 angstroms the nanometer wavelength of 237 which is much less sensitive for inactivation work for most microbes, in fact all of them, we find six fold the sensitivity which is still not very high of that where it should be in a DNA RNA system. No one has visualized the agents and not one but literally dozens of the major structural electron microscopists working on the subunit structure of viruses have been involved for decades in the attempt and their starting material are either tissue slices containing an infectivity tighter of 10 to the ninth per gram or zonal banding material even more rich in infectious particles. That they contain no host proteins I mentioned earlier. We doubt this but it looks as though it may be the case and attempts to immunize animals with material that have infectivity titers as high as 10 to the 11th per gram have produced no neutralizing, no radioimmune assay obtainable and low cytotoxic antibody that we can with any of the existing systems that work with retroviruses and all other groups of viruses that we can identify. Next slide please. The pathological lesion is a coalescence of vacuoles which have been studied and studied and studied their membrane is not very nice to look at it doesn't look there's no visible very virus like structure on it. Next slide. It expands and blow next slide please it blows up the cell and finally we end up with neuronal destruction. A good deal of implanted electrode physiology done at Percy Camard and Marseille with animals inoculated having previously had electrodes implanted has yielded evidence that for literally months and sometimes years before neuron death, neuron involvement starts and by controlled perfused pathological study this can be verified. The first proof that we had an infectious agent here came when chimpanzees succumbed to the clinical disease Kuru. A disease which produced a clinical picture which no behavioral analysis analysts had ever seen in zoo or naturally observed chimpanzees or in laboratory subhuman primates. In fact all the experimental animals develop a disease which has never been produced artifactually which has never had a similar pathological or clinical entity develop spontaneously of unknown or known cause in laboratory animals of the species used. So there is really no control necessary. The next slide the pathology of these animals is identical to that in man with the complex perkinje cell and molecular loss in the cerebellum I'm not going to expand the pathological discussions. The chimpanzee interestingly enough develops a drooping lip months before our clinical neurologist and veterinarians are aware that the disease is there. Our keepers by personality change and facial expression are aware of the disease long before hard neurological signs appear. Months before. This is true of course with Kuru in New Guinea and where even child patients recognize their fatal disorder before any physician or their parents are aware of it. Next slide. The analysis of cinema at a stage too early for our neurological astute colleagues to detect the disease does reveal restricted forelimb movement patterns as opposed to the normal walking pattern which coincides with the drooping lip and the keepers note of personality changes. Next slide. This added matter is of incredible interest if one realizes that during the two and four years of silent incubation in cats and in many species of old world monkeys when these animals would be said to be normal and when their neuropathologically studied brain by conventional means would be said to be normal. We now know that there are vast disturbances from implanted electrode work in thalmic, hypothalmic cortical relations disturbing paradoxical sleep and other sleep rhythm patterns which Juve is so interested in and this is happening with all the viruses. In addition we are finding if we use perfusion and early sacrifice neuronal nuclear amyototic division by nucleate and trinucleate neurons, fusion of neuron to neuron and neuron to glial cell as we now know these agents cause in vitro as well in tissue cultures. And we have a whole gamut of exciting pathology going on in the normal animal years before disease only beginning to ask the question does this ever go on in man. Since it's associated with only what we would consider allowable personality disturbances and sleep disturbances and it requires perfusion studies to study it is a difficult question to decide upon and one many of us are not anxious to have investigated that thoroughly. Now the Kuru patient who first produced chimpanzee disease took 21 months incubation when that brain material was placed in other chimpanzees it dropped to 11 months by 13 months in fact many this has been done over many times and again we get a drop in incubation from the usual one and a half to two years to less than a year to 13 or 14 months on first passage. This immediate drop on second passage to a shorter incubation period has been done over again over 60 times. We know it occurs and with other species the same one passage adoption adaptation looks like a selection process which we are not familiar with in virology. We are more used to a step by step slow adaptation to a new host. It doesn't proceed further on further passage. By the oral route there was no infection in this animal 12 years later is still well having had millions of lethal doses of the agent by the use of skin liver a clean liver kidney of the serial suspension without brain we still got the disease and using peripheral non-intra cerebral inoculation we got the disease very early in the work. I'm not going into all the passage studies. Next slide. This simply shows the first demonstration that we could shift from the chimpanzee to South American monkeys, new world monkeys with the sacrifice of a short incubation period of one year and making our waiting time into two years again. This has not reduced on further passage. Next slide. The new world monkeys, a dozen species of them, owl, spider, squirrel are all now used in work with the human diseases and it has permitted the curtailment of work with the diminishing rare species of chimpanzee which incident in our breeding colonies we breed more rapidly than we use. We had 30 newborn chimpanzees last year and only use five or six. We're doing more. Eventually we may be able to return them to Africa where they're being killed more frequently and indiscriminately. The passage in chimpanzee is not continuing. We're using smaller animals and other hosts now that we can. Next slide please. I'm not going into the experiments that that involved. This is a biopsy of a European patient with Kreuz-Valiacov disease to reemphasize the punched out spongiform change in the cortex. Next slide. To show you that we also found chimpanzee passage in 13 months of this disease possible and then in 12 and 40 months in another chimpanzee but almost three years and almost four years when we went to capuchin monkeys and marmosets. In other words these smaller monkeys require much more patience in the experimenter. Again when we found we could go to small animals and finally cats. We see chimpanzee work but one line has been continued. Next slide. Now to depart I'm going to not use the small remaining time to elaborate extensively on all the animal work that has been done and continues but to tell you a little about the worldwide form of Kuru. One sixth of the cases are familial. These patients had the disease. They weren't autopsyed but it's a clinically recognizable syndrome. So here are two aunts and mother of the propositist plus his grandmother all dying of Kreuz-Valiacov disease. Next slide please. Another family from Germany with three generations of many proved autopsyed cases from which the virus has been isolated. In fact these genealogies which are every bit as good for a dominant inheritance when they are looked at in a whole as for Huntington's Korea for instance. Number over 60 now in the number of families we know of the disease and from 11 of the first 17 we have had brain biopsy material on. We have already isolated the virus and the remaining six are not negative. They have not been inoculated long enough. Next slide. So here we have a disease which would undoubtedly be acceptable as a gene control to retopharmidial disease where a degenerative pathology is virus determined and with no genetic control that we know of in the fully susceptible hosts. Here is a huge amyloid plaque staining TAS positive, metachromatic, agentophilic. The plaque that all of us have in our brains as we age some people unfortunately have a great deal of it before 60 years of age and we call it Alzheimer's disease. This plaque is present in 20% of the Kreuzfeld patients. It's present in 60% of the Kuru patients, even children. But since it is not present in the remaining 40% of the Kuru patients, brothers, aunts, uncles, and parents of the other victims who have it, we are sure it's a host phenomenon that has nothing to do directly with pathogenesis. On the other hand, in Alzheimer's disease it and the tangles are what we most focus on and it is very remarkable that a fair number of Kreuzfeld patients have the disease. Many lead to pathological disputes as whether they are Alzheimer's or Kreuzfeld until we finally isolate the virus. Next slide. Simply demonstrating the spherical symmetry with cross nickel prisms of the amyloid which by EM is amyloid and by long chain and short chain analysis is amyloid. Next slide. To bring you up to date on what is just in press a world epidemiology in an American of Kreuzfeld, we now know that anywhere we look hard we find.5 cases per million per annum death. This makes it a very rare cause of death indeed. But when one realizes that this means 200 deaths a year for the United States and we have not had 10 deaths a year for rabies with a 50 state surveillance in the last 20 years, it's fully 50 times more frequent than rabies probably 100. This is a minimum. We are actually finding one per million per annum anywhere in large cities we look intensively. The European studies now impress with Katala and Paul Brown working on them in Paris for all of France, parts of Germany, most of Italy and Spain are showing.5 per million per annum on first survey for the last 10 years with 1 to 2 per million per annum in some foci and large intense foci in Hungary, Slovakia, Palma in Italy and in Israel. Next slide. Kreuzfeld is a worldwide disease. The countries of Africa where it's been found are few and far between. Hungary has not been intensive. We don't know why the, we haven't been to Argentina, the other countries in South America, everyone we visited, we found it and in high incidence. It far outrivals, outranks rabies as a world cause of death from infectious brain disease by probably a factor of 100 on all continents. Next slide. The scrappy virus is probably also ubiquitous where sheep are kept, but most countries with it do not recognize it. We have, and British workers have carried it for decades through mice and to our horror after long incubation period of 73 months the cinemologous monkey developed a spongiform encephalopathy identical with Kreuzfeld's disease. Their scrappy sheep brain inoculation as did after almost three years squirrel monkeys. On passage the incubation period has dropped in the squirrel monkey, but not in the cinemologous. Here it has been passed back into sheep and goat with disease proving that it is still the scrappy virus biologically, although since no Kreuzfeld or Kuru virus go this way. We are very worried however since European strains of scrappy taken from the Compton laboratories once transmitted to monkeys lose their ability to go back to sheep and goat. Conceivably scrappy is the source of some of the human Kreuzfeld. Next slide. It incidentally can be found in the butcher shops throughout the world. There's a high selection for the butcher of all scrappy infected sheep and even laboratory infected sheep throughout Europe having passed decades ended up in the butcher shop. Scrappy is a worldwide disease. Next slide. Now in a rush travelogue I promised you I will try to tell you where this story began. It began not in this enormous second largest island in the world but in one small highland area here in New Guinea. Next slide. The Kuru region is at 2000 to 3000 meters with human population scattered in the eastern highlands of a very mountainous area where all the grass covered areas represent the effects of millennia more than centuries of human cultivation by slash burn agriculture. Next slide. The villages are small few humans see or saw in the stone age period when we first described Kuru more than a few hundred individuals between their birth and their death in adult life. The population unit is under 100 in most places up to 200 in some and never exceeding that. Next slide please. This is a village in which we're returning the area has rugged and the first year of patrolling throughout the area required of moderate amount of mountaineering. Next slide and bridge building. Next slide. But the next slide I'd go through these rapidly and the mesolithic level culture of the population involved with Kuru are people who quite significantly never washed in a lifetime covering their body with the rendered grease from their pigs which run wild and mate with the wild pigs or of their relatives. Next slide. The kunai fields are high grass which long carrier line is carrying our supplies through. Next slide. And in that area we would meet one of the 11 language groups which were defined during the Kuru research project since only two of them had had European contact at the beginning of our Kuru epidemiological work. This is an Anga Kuka group which is just on the border. Next slide. The villages contain groups of women who were slow to leave as most women on our arrival. Preferred not to walk but when tested could walk but with minimal ataxia. They told us through the one and two level child translation we had into a language we could finally use that they were dying of Kuru and they were invariably right. The disease rarely lasted more than a year and was characterized by midline cerebellary ataxia with preservation of cortical functions and no sensory disturbance. Next slide. The patients at this stage are unable to focus their eyes or hold their neck properly so she looks demented but with grunting signals can evidence very good contact with reality and good intellect still. However she is so ataxic that she can no longer stand without support. Next slide. And in every village of 100 people we found people dying with no exception. Next slide. We found over 300 deaths the first year and in a given 100 population village three women using sticks to keep ambulating to their gardens to bring food back to their houses were already evidence that they had Kuru. They agreed that they did have no foray woman required to stick to maintain her balance unless she had Kuru or some other injury. Next slide. The smallest patients were children of four or five years of age. Next slide. And the area we found was of enormous linguistic diversity. The Nugini people, the Melanesian speak over 1000 languages not dialects like French, Spanish and Italian or German and Dutch. But real languages at a dialectical language it's much more diverse than that. In the eastern highlands alone there is this enormous level of languages and at least five language families. Next slide. As we worked out the boundaries of Kuru, next slide, we also gave names to most of the people which as is always the case with indigenous populations they are the fallacious names usually an insult like the bastards over the hill used by their enemy neighbors. The people themselves have no names for themselves. So our fallacious names have stuck and the new generation children don't know that we are the source of them. And this is the boundary of Kuru against the language groups that had it showing that here culture and language have determined the boundary completely. And here culture and language have had nothing to do with the boundary of the disease. Next slide. When we counted up cases at the end of a year and this prevalence data can be read as yearly mortality the disease usually lasts about a year. So this was 1.6% of the population dying per annum. This went on for 50 years previously and has gone on through most of the first 10 years of surveillance. But there's been a change. Next slide. Please. This simply the Kuru area would be superimposed on here and this is decreasing population density into high altitude mountain ranges and virgin bush with no people. Showing that the people became less and less dense as we went in this direction. Next slide. But the running away from Kuru which they consciously attributed their location in the virgin forests too was with no success whatsoever since maximum incidents with 3% dying per annum occurred in the least dense most remote populations bordering on populations here that had no disease at all. But there's no intermarriage or contact culturally between these groups. There's no ecological shift. Next slide. We began to accumulate patients. They were willing to come in until we had from 40 to 100 at one time dying in the O'Kapa hospital which we built. Dr. Zegas, for next slide. A New Guinea government physician who discovered the disease and has worked on it with me since the beginning. Is Vincent Zegas here myself in an early day with a child patient at O'Kapa? Next slide. And we would have this boy himself knows he's dying of Kuru but hadn't convinced us yet his autopsy six months later certainly proved he was right. He is supporting the girl with the disease and these two have the disease and that young girl does too. And next slide. The moon fascist is a result of unsuccessful attempts at cortisone therapy. Next slide. Next slide. It simply caught a patient during the movement. I should be showing you cinema not still pictures. We don't have the apparatus. Next slide please. And a severe strabismus and extreme dysarthria. A final total neuromuscular incapacity with minimal loss of intellectual function characterized all patients. Children usually died in six months to nine months. Next slide. Fully a third of all the patients were children and most of the patients were female except among children where the male to female ratio became a little more reasonable. This 13 to 1 to 78 to 1 ratio of female to male in adult life was very difficult to imagine epidemiologically at first. Next slide. We eventually found that from genealogical studies that a one of three wives, the people are polygamous and this enormous death of women had reduced the sex ratio to three to one raised it in favor of males in the southern villages for polygamous society with a total sex ratio of three to one. This made it closer to five to one for marriageable age males. Polygamy on top of it made it very hard for many young men. The every case with no exception of the 4,000 we have studied has had similar intense family history. Next slide. I'm not going to dwell on the usual family history. At present we know from the dating of these histories that this patient here was the contaminating source for the whole crowd but this has taken years of epidemiological sleuthing. I'm not going into it. Next slide. And here is what's happened. An exotic disease that few of us have had a chance to see and now we tell you it is almost gone before you've got to see it. It has disappeared progressively and the male and female patients have left us with only a few deaths. This was plotted prematurely. The final number in 77 actually was under 20 for the total. Next slide. This year it will be under 20. Next slide. But the most dramatic thing is that these are taken at three-year intervals, leaving out the intervening year in case of epidemiological end of year prejudice. It doesn't make much difference. It's a matter of Christmas New Year, European physicians doing less patrolling at that time. It shows that the zero to nine-year-old patients rapidly decreased, decreased and then disappeared from the world. The early adolescent patients rapidly decreased but later, five years later, then disappeared. Finally the older adolescent patients decreased and then disappeared. And now the 20 to 25-year-olds are gone. Every year the youngest patient is somewhat older. The law that no patient who was born since Kuru cannibalism stopped in his village has ever died of the disease holds. It tells us a great deal more than that first remark might immediately reveal. It tells us that the suspected transplacental transmission, milk factor transmission, vertical transmission from mother to child born during the disease or born before the mother is ill is not occurring. We're it occurring, this incredibly uniform and if I projected to 78, still more uniform with this mostly gone and that gone, disappearance would be unaccountable. Next slide. We do not have any such knowledge for the familial cases of Kreuzfelljacob in Germany and the rest of the world. If one doubts our ability to assess age of these now disappeared types of patients, pre-pubertyl children of both sexes, here is a group on dimer caprol and calcium-versonate therapy when we were looking for a Wilson's disease like copper or molybdenum toxicity and failed to find it. We had many false alarms in the beginning and all of them died of Kuru. There are no such patients in the last 10 years in the world. Next slide. The Kuru epidemiology was done mostly by the Robert Browning technique of the Pied Piper of Hamlet with assistance of here who are all married polygamous men today. Next slide, they rapidly, a group of some 30 of them provided, next slide provided us with translation of all 11 languages into the one language we learned. The young men who were leaving the area had to be initiated into adult life at which stage of life, all still pre-pubertyl, they have totally left the women's society never in their life even as polygamous adults to enter the house of a woman in traditional culture. All copulation and procreation was a daylight affair in the gardens where the privacy was greater than that obtained in the most remote Swiss valley and the phenomenon of sleeping with a wife is unknown to Highland New Guineaers until missionaries introduced this strange custom to them and the phenomenon of crawling in darkness for copulation under a roof is a phenomenon unheard of and rather repelling to them a subject of jest not to be taken seriously. Anointed with pig grease and never washed this would be human grease in a cannibal ceremony next slide but only for the smaller children and never the males. At this age people have left the women's culture only the women opened the Kuru victims bodies and contaminated themselves and children with them. Next slide they celebrated their initiation into manhood. Those three young men incidentally are all college graduates in the United States today. Next slide they are I'm not exaggerating. Next slide but this is the age at which all warfare took place most New Guinea Eastern Highland groups left that for the adolescent sex segregated culture while the old men as in most cultures planned the wars but didn't take part in them. Next slide and this age produced the sex segregation which did produce this enormous difference in sex predominance of the disease. Next slide girls at a early pubertal age were married with pig omentum on their head. Next slide and usually the young wife of a polygamous older man older widows went to the adolescents boys as their first wife. Next slide and all cooking was done over steam cooking with no vessels in pits of heated rock a meter and a half under this pile of several hundred pounds of pig meat and vegetables. It's an autoclave essentially a volcano like peep is earth is heaped on top of it and there is very little enteric disease since it comes right out of steam cooking. So the temperature attained would have been insufficient to inactivate the Kuru virus and human tissue was similarly handled. Next slide on the other hand with men handling pig tissue this would have been women for human tissue and the children around would have been infected. We believe the all infection was through mucus membranes and skin not orally it takes multi million lethal doses to infect with scraping orally only a millionth as much through skin or mucus membranes and with current cranes felt it doesn't take orally it is not eating the dead even though Claude Levy Strauss would have it totem wise symbolically that way. Next slide simply showing the children's involvement with the mumu pit of steam cooking. Next slide and finally the disappearance from the world of childhood Kuru ten years later the disappearance from the world of adolescent Kuru now the disappearance of young adult Kuru and hopefully within 20 years the disappearance from the world of Kuru how we will get rid of Kreuzfeld Jakob disease is another matter. We can tell you that only a few percent of the cases in the world five or six percent are attributable to the atrogenic cause. In Suric the use of stereotactic electrodes contaminated from a Kreuzfeld patient has produced two deaths years later in young people who were cured of their epilepsy by the surgery only to die from Kreuzfeld Jakob disease. Our form of neocannibalism brain use of in brain surgery of durameter from older patients and the use of corneal transplants from victims of Kreuzfeld Jakob disease has in fact caused the disease. Most major American and European clinics if thoroughly studied and if they have good enough records can detect the case caused by their neurosurgery. We do have a series of several dozen atrogenic cases the complex medical legal aspects of the issue and the humane aspects of the panic produced by epidemiological questioning of those in danger has restricted reporting of many of them. On the other hand we do already have the published accounts of a half dozen. One of the easiest ways of course to our utter horror of finding the virus is visiting a neuropsychiatric clinic where it would accumulate and we are just now aware of this hazard and changing the sterilization methods necessary to prevent the atrogenic spread since conventional sterilization with alcohol, ethylene oxide, sterilizers, zephyrin and many of these usual hospital antiseptics don't work. Because it smelled like hospital should in two, 30 years ago with Clorox, permanganate and iodine around which none of us and as young physicians want to take home to our families any longer on our clothes did inactivate the agents phenol did but the modern most of the modern antiseptics don't. There has been warning in all the world literature especially in the neuro and to pathologists neurosurgeons have died of the disease pathologists and dentists have we do not know and have no way of knowing whether the number that has died is a significant increase above what would be expected because of course they are closer to the medical profession and this rarely made difficult diagnosis is really even now missed in major academic neurological clinics as we are discovering in the worldwide epidemiology. I leave you then with a failure to have told you about the plural, the phenomenon that the Kuru focus is not the only intense focus of neurological or other chronic disease in primitive populations which we have pursued and are we are pursuing others, amyotrophic lateral sclerosis foci of 1000 to 2000 fold the intensity of any other civilized part of the world in West New Guinea populations and keep an ince of Japan and Guamani and Micronesian populations. Obviously if we ever find the answer to amyotrophic lateral sclerosis in these small enclaves which can be visited in a week where this has become the first cause of death in adult life are the place to find it. Thank you. Thank you.
 Our German hosts and fellow colleagues and students, I'm very grateful to be with you.  I'm coming directly from a much warmer climate about which I hope to speak to you a bit at  the end of my lecture.  And I'm suffering under the influence of your beautiful weather here.  The study of an epidemic of new disease, new to Western medicine, and a chronic, progressive,  always fatal central nervous system disease in a remote population isolate in people still  in a stone age culture in the central highlands of New Guinea, who, hate their own dead, has  a right of respect and mourning to their nearest kinsmen, has led us to two major discoveries.  The first, already mentioned by the over-generous introduction, was the discovery that chronic,  progressive, even at times totally non-inflammatory, and even apparently heredophomial diseases  could be caused by slow-acting virus infections, even years and at times several decades after  original infection and a long, silent incubation period.  The second discovery was that of a new group of microorganisms, which cause human diseases  and which possess such unusual physical, chemical, and biochemical, and biological properties  that many of our colleagues prefer that we not call them viruses.  They are so very different from all other microorganisms that they clearly represent  a new type of replicating agent, which we have tentatively decided to call unconventional  viruses.  The disease in question that led us to these discoveries is called Khuru, a now disappearing  plague of the Foray people and of ten other cultural and linguistic groups in the central  highlands of eastern New Guinea who intermarry with them.  They all inhabit a high mountainous interior of the eastern highlands, living at from 2,000  to 3,000 meters above sea level.  By opening the bodies and the skulls of their dead kinsmen, they contaminated the skin and  eyes principally of the women who performed the morning ritual and of the young children  with them.  This led to a fatal brain disease appearing 5, 10, even 20 or 30 years later.  It became the most common death among them.  In fact, it accounted in the first 10 years of our total surveillance of all the population  involved for over 50% of all death and among adult women, 90% of all those who died in the  15 year period of surveillance died from Khuru.  We have now discovered that the disease first described in Germany a half century ago and  named Kreuzfeld-Yakob type of presenial dementia after the two German neurologists who first  described it is a worldwide disease of at least tenfold and probably a hundredfold the  incidents that we attributed to it only a few years ago.  It is caused by a virus indistinguishable from that of Khuru, but whereas we know the mode  of transmission of Khuru from the contamination of the skin and mucous membranes through the  ritual rite of cannibalism, we do not know the usual means of transmission of our civilized  form of Khuru throughout the rest of the world.  We now know that these two unconventional viruses of man are similar to two such viruses  of animals, scrapey, tauberkrankheit of Deutsch or Tremblant en français and a disease called  transmissible mink encephalopathy of mink.  We call the group after the nature of its neuropathological lesion the subacute spongiform  virus encephalopathy.  Same resistance to an activation by ultraviolet radiation of 2,540 angstroms to 254 nanometers,  the wavelength which is the peak absorption wavelength for RNA and DNA has led to the  prediction that Khuru and CJD or Kreuzfeldt-Yakob disease virus and scrapey viruses possess  no nucleic acid.  This heresy has been further promulgated on the discovery that these unconventional viruses  also exhibit an enormous resistance to ionizing radiation using either gamma rays from a cobalt  60 source or neutron beam.  We're doing this work with Latigé at the Fondation Curie in Paris as well as at Oak  Ridge and we have repeated the work of our British colleagues, Hague and Tigger Alpers,  and confirm this resistance which is in the order of 100 to 1,000 times that known for  any other microbial system.  In fact the equivalent target size for a sphere with such unusual resistance to ionizing radiation  as specific, a non-specific, though the damage may be, would be calculated to be under 150,000  Dalton's, a molecular weight which is only one-tenth the size of the smallest genome  previously known to virology.  Finally, intensive study of these viruses for over a decade has failed to reveal any  antigenicity and in both the natural diseases and the experimentally induced diseases in  laboratory animals, there is no evidence that the victims develop any immunity or that the  immune system plays any role whatsoever in the disorder.  The humoral and the delayed hypersensitivity types of immune response, B cell and T cell  function remain intact and unstimulated.  There are no immune complexes formed and no immune complex deposits found on basement  glomerular membranes or in the chloride plexus.  These thus are the first mammalium microbial pathogens which apparently contain no non-host  protein in their structure.  Instead, they seem to be minute messenger RNA sized pieces of DNA tightly bound to host  membrane systems.  And indeed, the demonstration that they are in fact DNA is very tenuous based on only  one unconfirmed very recent experiment now in press.  The only structure we are left possible for them at present remains a tightly membrane  bound particle of size no larger than 150,000 Dalton's.  Being probably for none of its own enzymes or any core or coat proteins.  A virus without core or coat leaves those of us in structural virology most bewildered.  It is from recent work in plant virology that we found find sucre and the hope that we may  remain true to our basic tenets of our current DNA RNA religion and not have to retreat into  the previous speculations of replicating membranes devoid of nucleotide structures or replicating  basic proteins.  But I must warn you that the very same workers who with no difficulty whatsoever quickly  established the full sequence of FIX174 the smallest virus of bacteria yet sequenced have  been six years on this problem from Caltech and North Carolina out and in Germany and  in England and have failed yet to establish unequivocally that a nucleic acid is involved  in this whole system.  That we are dealing with viruses and viruses that cause human disease is very clear indeed.  They grow in vitro in tissue cultures of infected cells obtained by primary culture of the spleen,  liver, lymph nodes or brain of infected animals or primary human or animals with the natural  diseases.  Other cell lines such as L cells have been infected in vitro with the need of causing  fusion to get the infection started and the perhaps hazardous procedures of transforming  both human and animal cell lines with the SV40 genome so that they bear the tumor antigen  and still carry replicating simultaneously the infected genome of both the Croix-Velliacob  and the Coru agent and different cell lines and now the Scrapey have all been successfully  performed.  We are dealing with virus like agents.  We are dealing with a new group of microbes which might better be named by another term  and as usual the more fortunate plant virologists have preceded us and found that they must  also revise their basic microbial conceptions in a group of agents which are the smallest  of the plant viruses.  They prefer to call following dener the man who has defined them most thoroughly, viroids.  I will talk about them later and with that introduction I will start on a series of slides.  If I get through them on time I will then try to take you on a historical travelogue  to where this current day molecular biology began.  So if I pay somewhat short shrift to the rather lavish and hardly credible claims I have made  by beguipologies from my molecular biological colleagues to make it a little easier to follow  the terminology of a discipline of neurology and neuropathology notorious for the use of  impossible eponyms and impossible and almost meaningless descriptions based on neuropathological  lesion.  I will leave you rest assured that we have introduced only one new name into medicine  the name Kuru which is a nice non-anglo-saxon four letter word which has even crept into  the Webster Collegiate Dictionary whereas Lewis has yet failed.  I am unable to use any other term for Kreuzfeld-Yakob disease than the long established eponym.  The disease does run by 17 additional all longer synonyms in the neurological literature.  Scrape is a spontaneous disease of sheep and goats throughout the world and minkensophilopathy  we now have good reason both virologically and epidemiologically to believe is simply  the scrapy infection having spread to mink ranches naturally.  In fact it is no one that where it was first seen the carcasses of scrapy infected sheep  have been fed.  The next slide please.  This slide simply shows the natural diseases on the left the experimental diseases in experimentally  infected animals after over a year of incubation on the right.  It shows what is called status spongiosis or spongiform change by the neuropathologist  which simply means a sponge like punched out hole appearance of the gray matter it is not  referring to white matter spongiosis and although it was previously thought to be edema and  interstellular fluid accumulation it is exactly not that.  It is a swelling of neuronal and astroglyos cells based on a coalescence of minute vacuoles  which can be seen in their origin by electron microscopy long before they are visible to  light microscopists.  This is scrapy in sheep and scrapy in mice, curu in a child and curu in a chimpanzee inoculated  with the child's brain tissue.  Kreuzfeld Jacob from a brain biopsy in Oxford and the same process in the brain two years  later of a chimpanzee inoculated with it.  Next slide please.  The same distribution, scrapy, natural and experimental, curu natural and experimental,  Kreuzfeld natural and experimental demonstrating with a specific stain that shows astroglyos  cells the fibrous astroglyosis which is a dominant part of the pathological picture  and about as severe in this disease and this group of diseases as in any disease we know.  On the other hand these are the only infections of the brain and I make no exceptions in infections.  I mainly range through the whole range of such things as the protozoa through the viruses  in which there is no perivascular cuffing, no leukocytic invasion of the brain parenchyma  and no sign of a hypersensitivity or a inflammatory response.  Next slide.  For those clinically minded, next slide please.  The phenomenon that these diseases proceed from beginning to fatal outcome with no change  in spinal fluid protein and no pleocytosis at any stage of the disease is a further  unique finding in fatal CNS infections.  Now I'm not going to dwell on the hundreds of studies attempting to purify scrapy first  using cesium chloride then using calcium and potassium tartrate, metrismid as well as sucrose  and sucrose saline gradients have been used and the workers who have done this work are  not workers who have difficulty using zonal ultrace centrifuges and purifying reverse transcriptases,  viral subunits, P30 proteins, polyacrylamide gel, electrophoresing them but these same  workers have been unable to purify by these techniques or any other electrophoretic focusing  and the like the scrapy agent because no matter what small portion of maximum infectivity  at a density of 1.2 to 1.25 they take, it spreads again and electron microscopic control  of the re-zonal banding of it simply shows that we are trying to purify soap bubbles.  The material obtained with enormously high infectivity is a three-layered bioelectron  microscope plasma membrane light structure which fasciculates and the problem is that  of concentrating and purifying membranes where the reactive receptor or the reactive macromolecule  on the membrane is not known and although we assume it is a small nucleic acid all attempts  with hot formaldehyde and with detergents to release this presumed genome have failed  to obtain any infectious nucleic acid.  Next slide.  The most disturbing early data which came first from Compton working on scrapy then we  reconfirmed it in using zonal banded and fluorocarbon cleaned brain suspended scrapy virus.  We obtained linear inactivation curves at ergs 10 to the per square millimeter times  10 to the fourth.  A energy input which if you are not familiar with radiation work would not lead you to  realize that the most radio resistant forms of life we know to 254 nanometer radiation  would intersect here at six.  They would be the U1 variant of tobacco mosaic and bacillus radiodurans.  This was interpreted first as absolute proof that the agents could not contain nucleic  acid.  In the suspensions if the argument is that they are dirty not sufficiently cleaned one  puts a large variety of mammalian plant viruses and bacteriophage.  These so called dirty suspensions have no effect on protection at this radiation.  Next slide.  I am not going to dwell on the long discussion this might entail but here is the same data  you just saw but it simply shows that the plant extracts of the plant viroids these  newly described smallest known viruses are even more radio resistant and the purified  RNA genome of 115,000 molecular weight of potato spindle tuber virus is of the same  order of magnitude of radio resistance whereas all conventional viruses fall here.  Next slide please.  This alone eliminates all other plant and animal viruses from consideration.  Studies of very unsatisfactory zonal banding into 55 fractions in a sucrose saline gradient  that the best anyone has obtained is a log scale so it doesn't plot out as well as the  earlier ones I showed you.  I have failed to reveal any relation with marker enzyme systems in enzyme activity for  three lysosomal membrane bound enzymes and one mitochondrial enzyme.  Next slide please.  I am now going to rapidly summarize probably as fast as you can read.  Much of what I have said simply to drive home the fact that these surely are an unconventional  type of viral agent.  On the physical chemical side I fail to mention what alone would leave any earlier microbiologist  totally skeptical of the integrity of the workers.  That a virus or any microbe can be stored in 4% formaldehyde or 10 or 20% formaldehyde  at room temperature for years without losing tighter namely embalming fluid and that which  we use in pathological laboratories to store specimens is blatantly ridiculous.  It happens to be the fact.  We have isolated Creutz-Feliaco virus from material that has been stored years at room  temperature in pathological laboratories in formaldehyde.  Scrapey and minkencephalopathy have been taken off paraffin embedded blocks which were for  two years in formaldehyde before they were sectioned and left several years as blocks  and obtained as viable microbes.  Beta-propreolactone doesn't touch them.  Ethylene, diamine, tetraacetic acid doesn't touch them.  Proteases and the nucleases on even the deprotonized fluorocarbon treated zonal band infractions  are not effective nor can we use hot formaldehyde to inactivate them and searching for double  or single stranded RNA.  Heat resistance is not incompatible with the biological experience but it is incompatible  with most virological experience.  Up until 80 degrees we have full stability.  At 85 degrees we begin to inactivate and as with the hepatitis virus, boiling is not  sufficient for hospital sterilization.  Autoclaving however most certainly is.  Next slide please.  To go on further I have already summarized the UV resistance for you.  I've mentioned the enormous ionizing radiation resistance.  We use ultrasonic energy in such energy input that we would be inactivating rapidly polio  or flu or most conventional viruses and all we do is gain in yield increasing our infectivity  tighter.  When we do an action spectrum at 2,370 angstroms the nanometer wavelength of 237 which is much  less sensitive for inactivation work for most microbes, in fact all of them, we find six  fold the sensitivity which is still not very high of that where it should be in a DNA RNA  system.  No one has visualized the agents and not one but literally dozens of the major structural  electron microscopists working on the subunit structure of viruses have been involved for  decades in the attempt and their starting material are either tissue slices containing  an infectivity tighter of 10 to the ninth per gram or zonal banding material even more  rich in infectious particles.  That they contain no host proteins I mentioned earlier.  We doubt this but it looks as though it may be the case and attempts to immunize animals  with material that have infectivity titers as high as 10 to the 11th per gram have produced  no neutralizing, no radioimmune assay obtainable and low cytotoxic antibody that we can with  any of the existing systems that work with retroviruses and all other groups of viruses  that we can identify.  Next slide please.  The pathological lesion is a coalescence of vacuoles which have been studied and studied  and studied their membrane is not very nice to look at it doesn't look there's no visible  very virus like structure on it.  Next slide.  It expands and blow next slide please it blows up the cell and finally we end up with neuronal  destruction.  A good deal of implanted electrode physiology done at Percy Camard and Marseille with animals  inoculated having previously had electrodes implanted has yielded evidence that for literally  months and sometimes years before neuron death, neuron involvement starts and by controlled  perfused pathological study this can be verified.  The first proof that we had an infectious agent here came when chimpanzees succumbed  to the clinical disease Kuru.  A disease which produced a clinical picture which no behavioral analysis analysts had  ever seen in zoo or naturally observed chimpanzees or in laboratory subhuman primates.  In fact all the experimental animals develop a disease which has never been produced artifactually  which has never had a similar pathological or clinical entity develop spontaneously of  unknown or known cause in laboratory animals of the species used.  So there is really no control necessary.  The next slide the pathology of these animals is identical to that in man with the complex  perkinje cell and molecular loss in the cerebellum I'm not going to expand the pathological discussions.  The chimpanzee interestingly enough develops a drooping lip months before our clinical  neurologist and veterinarians are aware that the disease is there.  Our keepers by personality change and facial expression are aware of the disease long before  hard neurological signs appear.  Months before.  This is true of course with Kuru in New Guinea and where even child patients recognize their  fatal disorder before any physician or their parents are aware of it.  Next slide.  The analysis of cinema at a stage too early for our neurological astute colleagues to  detect the disease does reveal restricted forelimb movement patterns as opposed to the normal  walking pattern which coincides with the drooping lip and the keepers note of personality changes.  Next slide.  This added matter is of incredible interest if one realizes that during the two and four  years of silent incubation in cats and in many species of old world monkeys when these  animals would be said to be normal and when their neuropathologically studied brain by  conventional means would be said to be normal.  We now know that there are vast disturbances from implanted electrode work in thalmic,  hypothalmic cortical relations disturbing paradoxical sleep and other sleep rhythm patterns which  Juve is so interested in and this is happening with all the viruses.  In addition we are finding if we use perfusion and early sacrifice neuronal nuclear amyototic  division by nucleate and trinucleate neurons, fusion of neuron to neuron and neuron to glial  cell as we now know these agents cause in vitro as well in tissue cultures.  And we have a whole gamut of exciting pathology going on in the normal animal years before  disease only beginning to ask the question does this ever go on in man.  Since it's associated with only what we would consider allowable personality disturbances  and sleep disturbances and it requires perfusion studies to study it is a difficult question  to decide upon and one many of us are not anxious to have investigated that thoroughly.  Now the Kuru patient who first produced chimpanzee disease took 21 months incubation when that  brain material was placed in other chimpanzees it dropped to 11 months by 13 months in fact  many this has been done over many times and again we get a drop in incubation from the  usual one and a half to two years to less than a year to 13 or 14 months on first passage.  This immediate drop on second passage to a shorter incubation period has been done over  again over 60 times.  We know it occurs and with other species the same one passage adoption adaptation looks  like a selection process which we are not familiar with in virology.  We are more used to a step by step slow adaptation to a new host.  It doesn't proceed further on further passage.  By the oral route there was no infection in this animal 12 years later is still well having  had millions of lethal doses of the agent by the use of skin liver a clean liver kidney  of the serial suspension without brain we still got the disease and using peripheral  non-intra cerebral inoculation we got the disease very early in the work.  I'm not going into all the passage studies.  Next slide.  This simply shows the first demonstration that we could shift from the chimpanzee to  South American monkeys, new world monkeys with the sacrifice of a short incubation period  of one year and making our waiting time into two years again.  This has not reduced on further passage.  Next slide.  The new world monkeys, a dozen species of them, owl, spider, squirrel are all now used  in work with the human diseases and it has permitted the curtailment of work with the  diminishing rare species of chimpanzee which incident in our breeding colonies we breed  more rapidly than we use.  We had 30 newborn chimpanzees last year and only use five or six.  We're doing more.  Eventually we may be able to return them to Africa where they're being killed more frequently  and indiscriminately.  The passage in chimpanzee is not continuing.  We're using smaller animals and other hosts now that we can.  Next slide please.  I'm not going into the experiments that that involved.  This is a biopsy of a European patient with Kreuz-Valiacov disease to reemphasize the  punched out spongiform change in the cortex.  Next slide.  To show you that we also found chimpanzee passage in 13 months of this disease possible  and then in 12 and 40 months in another chimpanzee but almost three years and almost four years  when we went to capuchin monkeys and marmosets.  In other words these smaller monkeys require much more patience in the experimenter.  Again when we found we could go to small animals and finally cats.  We see chimpanzee work but one line has been continued.  Next slide.  Now to depart I'm going to not use the small remaining time to elaborate extensively on  all the animal work that has been done and continues but to tell you a little about the  worldwide form of Kuru.  One sixth of the cases are familial.  These patients had the disease.  They weren't autopsyed but it's a clinically recognizable syndrome.  So here are two aunts and mother of the propositist plus his grandmother all dying of Kreuz-Valiacov  disease.  Next slide please.  Another family from Germany with three generations of many proved autopsyed cases from which  the virus has been isolated.  In fact these genealogies which are every bit as good for a dominant inheritance when  they are looked at in a whole as for Huntington's Korea for instance.  Number over 60 now in the number of families we know of the disease and from 11 of the  first 17 we have had brain biopsy material on.  We have already isolated the virus and the remaining six are not negative.  They have not been inoculated long enough.  Next slide.  So here we have a disease which would undoubtedly be acceptable as a gene control to retopharmidial  disease where a degenerative pathology is virus determined and with no genetic control  that we know of in the fully susceptible hosts.  Here is a huge amyloid plaque staining TAS positive, metachromatic, agentophilic.  The plaque that all of us have in our brains as we age some people unfortunately have a  great deal of it before 60 years of age and we call it Alzheimer's disease.  This plaque is present in 20% of the Kreuzfeld patients.  It's present in 60% of the Kuru patients, even children.  But since it is not present in the remaining 40% of the Kuru patients, brothers, aunts,  uncles, and parents of the other victims who have it, we are sure it's a host phenomenon  that has nothing to do directly with pathogenesis.  On the other hand, in Alzheimer's disease it and the tangles are what we most focus  on and it is very remarkable that a fair number of Kreuzfeld patients have the disease.  Many lead to pathological disputes as whether they are Alzheimer's or Kreuzfeld until we  finally isolate the virus.  Next slide.  Simply demonstrating the spherical symmetry with cross nickel prisms of the amyloid which  by EM is amyloid and by long chain and short chain analysis is amyloid.  Next slide.  To bring you up to date on what is just in press a world epidemiology in an American  of Kreuzfeld, we now know that anywhere we look hard we find.5 cases per million per  annum death.  This makes it a very rare cause of death indeed.  But when one realizes that this means 200 deaths a year for the United States and we  have not had 10 deaths a year for rabies with a 50 state surveillance in the last 20 years,  it's fully 50 times more frequent than rabies probably 100.  This is a minimum.  We are actually finding one per million per annum anywhere in large cities we look intensively.  The European studies now impress with Katala and Paul Brown working on them in Paris for  all of France, parts of Germany, most of Italy and Spain are showing.5 per million per  annum on first survey for the last 10 years with 1 to 2 per million per annum in some  foci and large intense foci in Hungary, Slovakia, Palma in Italy and in Israel.  Next slide.  Kreuzfeld is a worldwide disease.  The countries of Africa where it's been found are few and far between.  Hungary has not been intensive.  We don't know why the, we haven't been to Argentina, the other countries in South America, everyone  we visited, we found it and in high incidence.  It far outrivals, outranks rabies as a world cause of death from infectious brain disease  by probably a factor of 100 on all continents.  Next slide.  The scrappy virus is probably also ubiquitous where sheep are kept, but most countries with  it do not recognize it.  We have, and British workers have carried it for decades through mice and to our horror  after long incubation period of 73 months the cinemologous monkey developed a spongiform  encephalopathy identical with Kreuzfeld's disease.  Their scrappy sheep brain inoculation as did after almost three years squirrel monkeys.  On passage the incubation period has dropped in the squirrel monkey, but not in the cinemologous.  Here it has been passed back into sheep and goat with disease proving that it is still  the scrappy virus biologically, although since no Kreuzfeld or Kuru virus go this way.  We are very worried however since European strains of scrappy taken from the Compton laboratories  once transmitted to monkeys lose their ability to go back to sheep and goat.  Conceivably scrappy is the source of some of the human Kreuzfeld.  Next slide.  It incidentally can be found in the butcher shops throughout the world.  There's a high selection for the butcher of all scrappy infected sheep and even laboratory  infected sheep throughout Europe having passed decades ended up in the butcher shop.  Scrappy is a worldwide disease.  Next slide.  Now in a rush travelogue I promised you I will try to tell you where this story began.  It began not in this enormous second largest island in the world but in one small highland  area here in New Guinea.  Next slide. The Kuru region is at 2000 to 3000 meters with human population scattered  in the eastern highlands of a very mountainous area where all the grass covered areas represent  the effects of millennia more than centuries of human cultivation by slash burn agriculture.  Next slide.  The villages are small few humans see or saw in the stone age period when we first described  Kuru more than a few hundred individuals between their birth and their death in adult life.  The population unit is under 100 in most places up to 200 in some and never exceeding that.  Next slide please.  This is a village in which we're returning the area has rugged and the first year of  patrolling throughout the area required of moderate amount of mountaineering.  Next slide and bridge building.  Next slide.  But the next slide I'd go through these rapidly and the mesolithic level culture of the population  involved with Kuru are people who quite significantly never washed in a lifetime covering their body  with the rendered grease from their pigs which run wild and mate with the wild pigs or  of their relatives.  Next slide.  The kunai fields are high grass which long carrier line is carrying our supplies through.  Next slide.  And in that area we would meet one of the 11 language groups which were defined during  the Kuru research project since only two of them had had European contact at the beginning  of our Kuru epidemiological work.  This is an Anga Kuka group which is just on the border.  Next slide.  The villages contain groups of women who were slow to leave as most women on our arrival.  Preferred not to walk but when tested could walk but with minimal ataxia.  They told us through the one and two level child translation we had into a language we  could finally use that they were dying of Kuru and they were invariably right.  The disease rarely lasted more than a year and was characterized by midline cerebellary  ataxia with preservation of cortical functions and no sensory disturbance.  Next slide.  The patients at this stage are unable to focus their eyes or hold their neck properly so  she looks demented but with grunting signals can evidence very good contact with reality  and good intellect still.  However she is so ataxic that she can no longer stand without support.  Next slide.  And in every village of 100 people we found people dying with no exception.  Next slide.  We found over 300 deaths the first year and in a given 100 population village three women  using sticks to keep ambulating to their gardens to bring food back to their houses were already  evidence that they had Kuru.  They agreed that they did have no foray woman required to stick to maintain her balance  unless she had Kuru or some other injury.  Next slide.  The smallest patients were children of four or five years of age.  Next slide.  And the area we found was of enormous linguistic diversity.  The Nugini people, the Melanesian speak over 1000 languages not dialects like French, Spanish  and Italian or German and Dutch.  But real languages at a dialectical language it's much more diverse than that.  In the eastern highlands alone there is this enormous level of languages and at least five  language families.  Next slide.  As we worked out the boundaries of Kuru, next slide, we also gave names to most of the  people which as is always the case with indigenous populations they are the fallacious names  usually an insult like the bastards over the hill used by their enemy neighbors.  The people themselves have no names for themselves.  So our fallacious names have stuck and the new generation children don't know that we  are the source of them.  And this is the boundary of Kuru against the language groups that had it showing that here  culture and language have determined the boundary completely.  And here culture and language have had nothing to do with the boundary of the disease.  Next slide.  When we counted up cases at the end of a year and this prevalence data can be read as yearly  mortality the disease usually lasts about a year.  So this was 1.6% of the population dying per annum.  This went on for 50 years previously and has gone on through most of the first 10 years  of surveillance.  But there's been a change.  Next slide.  Please.  This simply the Kuru area would be superimposed on here and this is decreasing population  density into high altitude mountain ranges and virgin bush with no people.  Showing that the people became less and less dense as we went in this direction.  Next slide.  But the running away from Kuru which they consciously attributed their location in the  virgin forests too was with no success whatsoever since maximum incidents with 3% dying per  annum occurred in the least dense most remote populations bordering on populations here  that had no disease at all.  But there's no intermarriage or contact culturally between these groups.  There's no ecological shift.  Next slide.  We began to accumulate patients.  They were willing to come in until we had from 40 to 100 at one time dying in the O'Kapa  hospital which we built.  Dr. Zegas, for next slide.  A New Guinea government physician who discovered the disease and has worked on it with me since  the beginning.  Is Vincent Zegas here myself in an early day with a child patient at O'Kapa?  Next slide.  And we would have this boy himself knows he's dying of Kuru but hadn't convinced us  yet his autopsy six months later certainly proved he was right.  He is supporting the girl with the disease and these two have the disease and that young  girl does too.  And next slide.  The moon fascist is a result of unsuccessful attempts at cortisone therapy.  Next slide.  Next slide.  It simply caught a patient during the movement.  I should be showing you cinema not still pictures.  We don't have the apparatus.  Next slide please.  And a severe strabismus and extreme dysarthria.  A final total neuromuscular incapacity with minimal loss of intellectual function characterized  all patients.  Children usually died in six months to nine months.  Next slide.  Fully a third of all the patients were children and most of the patients were female except  among children where the male to female ratio became a little more reasonable.  This 13 to 1 to 78 to 1 ratio of female to male in adult life was very difficult to imagine  epidemiologically at first.  Next slide.  We eventually found that from genealogical studies that a one of three wives, the people  are polygamous and this enormous death of women had reduced the sex ratio to three to  one raised it in favor of males in the southern villages for polygamous society with a total  sex ratio of three to one.  This made it closer to five to one for marriageable age males.  Polygamy on top of it made it very hard for many young men.  The every case with no exception of the 4,000 we have studied has had similar intense family  history.  Next slide.  I'm not going to dwell on the usual family history.  At present we know from the dating of these histories that this patient here was the contaminating  source for the whole crowd but this has taken years of epidemiological sleuthing.  I'm not going into it.  Next slide.  And here is what's happened.  An exotic disease that few of us have had a chance to see and now we tell you it is almost  gone before you've got to see it.  It has disappeared progressively and the male and female patients have left us with only  a few deaths.  This was plotted prematurely.  The final number in 77 actually was under 20 for the total.  Next slide.  This year it will be under 20.  Next slide.  But the most dramatic thing is that these are taken at three-year intervals, leaving  out the intervening year in case of epidemiological end of year prejudice.  It doesn't make much difference.  It's a matter of Christmas New Year, European physicians doing less patrolling at that time.  It shows that the zero to nine-year-old patients rapidly decreased, decreased and then disappeared  from the world.  The early adolescent patients rapidly decreased but later, five years later, then disappeared.  Finally the older adolescent patients decreased and then disappeared.  And now the 20 to 25-year-olds are gone.  Every year the youngest patient is somewhat older.  The law that no patient who was born since Kuru cannibalism stopped in his village has  ever died of the disease holds.  It tells us a great deal more than that first remark might immediately reveal.  It tells us that the suspected transplacental transmission, milk factor transmission, vertical  transmission from mother to child born during the disease or born before the mother is ill  is not occurring.  We're it occurring, this incredibly uniform and if I projected to 78, still more uniform  with this mostly gone and that gone, disappearance would be unaccountable.  Next slide.  We do not have any such knowledge for the familial cases of Kreuzfelljacob in Germany  and the rest of the world.  If one doubts our ability to assess age of these now disappeared types of patients, pre-pubertyl  children of both sexes, here is a group on dimer caprol and calcium-versonate therapy  when we were looking for a Wilson's disease like copper or molybdenum toxicity and failed  to find it.  We had many false alarms in the beginning and all of them died of Kuru.  There are no such patients in the last 10 years in the world.  Next slide.  The Kuru epidemiology was done mostly by the Robert Browning technique of the Pied Piper  of Hamlet with assistance of here who are all married polygamous men today.  Next slide, they rapidly, a group of some 30 of them provided, next slide provided us  with translation of all 11 languages into the one language we learned.  The young men who were leaving the area had to be initiated into adult life at which stage  of life, all still pre-pubertyl, they have totally left the women's society never in  their life even as polygamous adults to enter the house of a woman in traditional culture.  All copulation and procreation was a daylight affair in the gardens where the privacy was  greater than that obtained in the most remote Swiss valley and the phenomenon of sleeping  with a wife is unknown to Highland New Guineaers until missionaries introduced this strange  custom to them and the phenomenon of crawling in darkness for copulation under a roof is  a phenomenon unheard of and rather repelling to them a subject of jest not to be taken  seriously.  Anointed with pig grease and never washed this would be human grease in a cannibal  ceremony next slide but only for the smaller children and never the males.  At this age people have left the women's culture only the women opened the Kuru victims bodies  and contaminated themselves and children with them.  Next slide they celebrated their initiation into manhood.  Those three young men incidentally are all college graduates in the United States today.  Next slide they are I'm not exaggerating.  Next slide but this is the age at which all warfare took place most New Guinea Eastern  Highland groups left that for the adolescent sex segregated culture while the old men as  in most cultures planned the wars but didn't take part in them.  Next slide and this age produced the sex segregation which did produce this enormous difference  in sex predominance of the disease.  Next slide girls at a early pubertal age were married with pig omentum on their head.  Next slide and usually the young wife of a polygamous older man older widows went to  the adolescents boys as their first wife.  Next slide and all cooking was done over steam cooking with no vessels in pits of heated  rock a meter and a half under this pile of several hundred pounds of pig meat and vegetables.  It's an autoclave essentially a volcano like peep is earth is heaped on top of it and there  is very little enteric disease since it comes right out of steam cooking.  So the temperature attained would have been insufficient to inactivate the Kuru virus  and human tissue was similarly handled.  Next slide on the other hand with men handling pig tissue this would have been women for  human tissue and the children around would have been infected.  We believe the all infection was through mucus membranes and skin not orally it takes multi  million lethal doses to infect with scraping orally only a millionth as much through skin  or mucus membranes and with current cranes felt it doesn't take orally it is not eating  the dead even though Claude Levy Strauss would have it totem wise symbolically that way.  Next slide simply showing the children's involvement with the mumu pit of steam cooking.  Next slide and finally the disappearance from the world of childhood Kuru ten years later  the disappearance from the world of adolescent Kuru now the disappearance of young adult  Kuru and hopefully within 20 years the disappearance from the world of Kuru how we will get rid  of Kreuzfeld Jakob disease is another matter.  We can tell you that only a few percent of the cases in the world five or six percent  are attributable to the atrogenic cause.  In Suric the use of stereotactic electrodes contaminated from a Kreuzfeld patient has  produced two deaths years later in young people who were cured of their epilepsy by the surgery  only to die from Kreuzfeld Jakob disease.  Our form of neocannibalism brain use of in brain surgery of durameter from older patients  and the use of corneal transplants from victims of Kreuzfeld Jakob disease has in fact caused  the disease.  Most major American and European clinics if thoroughly studied and if they have good enough  records can detect the case caused by their neurosurgery.  We do have a series of several dozen atrogenic cases the complex medical legal aspects of  the issue and the humane aspects of the panic produced by epidemiological questioning of  those in danger has restricted reporting of many of them.  On the other hand we do already have the published accounts of a half dozen.  One of the easiest ways of course to our utter horror of finding the virus is visiting a  neuropsychiatric clinic where it would accumulate and we are just now aware of this hazard and  changing the sterilization methods necessary to prevent the atrogenic spread since conventional  sterilization with alcohol, ethylene oxide, sterilizers, zephyrin and many of these usual  hospital antiseptics don't work.  Because it smelled like hospital should in two, 30 years ago with Clorox, permanganate  and iodine around which none of us and as young physicians want to take home to our  families any longer on our clothes did inactivate the agents phenol did but the modern most  of the modern antiseptics don't.  There has been warning in all the world literature especially in the neuro and to pathologists  neurosurgeons have died of the disease pathologists and dentists have we do not know and have  no way of knowing whether the number that has died is a significant increase above what  would be expected because of course they are closer to the medical profession and this  rarely made difficult diagnosis is really even now missed in major academic neurological  clinics as we are discovering in the worldwide epidemiology.  I leave you then with a failure to have told you about the plural, the phenomenon that the  Kuru focus is not the only intense focus of neurological or other chronic disease in  primitive populations which we have pursued and are we are pursuing others, amyotrophic  lateral sclerosis foci of 1000 to 2000 fold the intensity of any other civilized part  of the world in West New Guinea populations and keep an ince of Japan and Guamani and  Micronesian populations.  Obviously if we ever find the answer to amyotrophic lateral sclerosis in these small enclaves  which can be visited in a week where this has become the first cause of death in adult  life are the place to find it.  Thank you.  Thank you.
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Our German hosts and fellow colleagues and students, I'm very grateful to be with you. I'm coming directly from a much warmer climate about which I hope to speak to you a bit at the end of my lecture. And I'm suffering under the influence of your beautiful weather here. The study of an epidemic of new disease, new to Western medicine, and a chronic, progressive, always fatal central nervous system disease in a remote population isolate in people still in a stone age culture in the central highlands of New Guinea, who, hate their own dead, has a right of respect and mourning to their nearest kinsmen, has led us to two major discoveries. The first, already mentioned by the over-generous introduction, was the discovery that chronic, progressive, even at times totally non-inflammatory, and even apparently heredophomial diseases could be caused by slow-acting virus infections, even years and at times several decades after original infection and a long, silent incubation period. The second discovery was that of a new group of microorganisms, which cause human diseases and which possess such unusual physical, chemical, and biochemical, and biological properties that many of our colleagues prefer that we not call them viruses. They are so very different from all other microorganisms that they clearly represent a new type of replicating agent, which we have tentatively decided to call unconventional viruses. The disease in question that led us to these discoveries is called Khuru, a now disappearing plague of the Foray people and of ten other cultural and linguistic groups in the central highlands of eastern New Guinea who intermarry with them. They all inhabit a high mountainous interior of the eastern highlands, living at from 2,000 to 3,000 meters above sea level. By opening the bodies and the skulls of their dead kinsmen, they contaminated the skin and eyes principally of the women who performed the morning ritual and of the young children with them. This led to a fatal brain disease appearing 5, 10, even 20 or 30 years later. It became the most common death among them. In fact, it accounted in the first 10 years of our total surveillance of all the population involved for over 50% of all death and among adult women, 90% of all those who died in the 15 year period of surveillance died from Khuru. We have now discovered that the disease first described in Germany a half century ago and named Kreuzfeld-Yakob type of presenial dementia after the two German neurologists who first described it is a worldwide disease of at least tenfold and probably a hundredfold the incidents that we attributed to it only a few years ago. It is caused by a virus indistinguishable from that of Khuru, but whereas we know the mode of transmission of Khuru from the contamination of the skin and mucous membranes through the ritual rite of cannibalism, we do not know the usual means of transmission of our civilized form of Khuru throughout the rest of the world. We now know that these two unconventional viruses of man are similar to two such viruses of animals, scrapey, tauberkrankheit of Deutsch or Tremblant en français and a disease called transmissible mink encephalopathy of mink. We call the group after the nature of its neuropathological lesion the subacute spongiform virus encephalopathy. Same resistance to an activation by ultraviolet radiation of 2,540 angstroms to 254 nanometers, the wavelength which is the peak absorption wavelength for RNA and DNA has led to the prediction that Khuru and CJD or Kreuzfeldt-Yakob disease virus and scrapey viruses possess no nucleic acid. This heresy has been further promulgated on the discovery that these unconventional viruses also exhibit an enormous resistance to ionizing radiation using either gamma rays from a cobalt 60 source or neutron beam. We're doing this work with Latigé at the Fondation Curie in Paris as well as at Oak Ridge and we have repeated the work of our British colleagues, Hague and Tigger Alpers, and confirm this resistance which is in the order of 100 to 1,000 times that known for any other microbial system. In fact the equivalent target size for a sphere with such unusual resistance to ionizing radiation as specific, a non-specific, though the damage may be, would be calculated to be under 150,000 Dalton's, a molecular weight which is only one-tenth the size of the smallest genome previously known to virology. Finally, intensive study of these viruses for over a decade has failed to reveal any antigenicity and in both the natural diseases and the experimentally induced diseases in laboratory animals, there is no evidence that the victims develop any immunity or that the immune system plays any role whatsoever in the disorder. The humoral and the delayed hypersensitivity types of immune response, B cell and T cell function remain intact and unstimulated. There are no immune complexes formed and no immune complex deposits found on basement glomerular membranes or in the chloride plexus. These thus are the first mammalium microbial pathogens which apparently contain no non-host protein in their structure. Instead, they seem to be minute messenger RNA sized pieces of DNA tightly bound to host membrane systems. And indeed, the demonstration that they are in fact DNA is very tenuous based on only one unconfirmed very recent experiment now in press. The only structure we are left possible for them at present remains a tightly membrane bound particle of size no larger than 150,000 Dalton's. Being probably for none of its own enzymes or any core or coat proteins. A virus without core or coat leaves those of us in structural virology most bewildered. It is from recent work in plant virology that we found find sucre and the hope that we may remain true to our basic tenets of our current DNA RNA religion and not have to retreat into the previous speculations of replicating membranes devoid of nucleotide structures or replicating basic proteins. But I must warn you that the very same workers who with no difficulty whatsoever quickly established the full sequence of FIX174 the smallest virus of bacteria yet sequenced have been six years on this problem from Caltech and North Carolina out and in Germany and in England and have failed yet to establish unequivocally that a nucleic acid is involved in this whole system. That we are dealing with viruses and viruses that cause human disease is very clear indeed. They grow in vitro in tissue cultures of infected cells obtained by primary culture of the spleen, liver, lymph nodes or brain of infected animals or primary human or animals with the natural diseases. Other cell lines such as L cells have been infected in vitro with the need of causing fusion to get the infection started and the perhaps hazardous procedures of transforming both human and animal cell lines with the SV40 genome so that they bear the tumor antigen and still carry replicating simultaneously the infected genome of both the Croix-Velliacob and the Coru agent and different cell lines and now the Scrapey have all been successfully performed. We are dealing with virus like agents. We are dealing with a new group of microbes which might better be named by another term and as usual the more fortunate plant virologists have preceded us and found that they must also revise their basic microbial conceptions in a group of agents which are the smallest of the plant viruses. They prefer to call following dener the man who has defined them most thoroughly, viroids. I will talk about them later and with that introduction I will start on a series of slides. If I get through them on time I will then try to take you on a historical travelogue to where this current day molecular biology began. So if I pay somewhat short shrift to the rather lavish and hardly credible claims I have made by beguipologies from my molecular biological colleagues to make it a little easier to follow the terminology of a discipline of neurology and neuropathology notorious for the use of impossible eponyms and impossible and almost meaningless descriptions based on neuropathological lesion. I will leave you rest assured that we have introduced only one new name into medicine the name Kuru which is a nice non-anglo-saxon four letter word which has even crept into the Webster Collegiate Dictionary whereas Lewis has yet failed. I am unable to use any other term for Kreuzfeld-Yakob disease than the long established eponym. The disease does run by 17 additional all longer synonyms in the neurological literature. Scrape is a spontaneous disease of sheep and goats throughout the world and minkensophilopathy we now have good reason both virologically and epidemiologically to believe is simply the scrapy infection having spread to mink ranches naturally. In fact it is no one that where it was first seen the carcasses of scrapy infected sheep have been fed. The next slide please. This slide simply shows the natural diseases on the left the experimental diseases in experimentally infected animals after over a year of incubation on the right. It shows what is called status spongiosis or spongiform change by the neuropathologist which simply means a sponge like punched out hole appearance of the gray matter it is not referring to white matter spongiosis and although it was previously thought to be edema and interstellular fluid accumulation it is exactly not that. It is a swelling of neuronal and astroglyos cells based on a coalescence of minute vacuoles which can be seen in their origin by electron microscopy long before they are visible to light microscopists. This is scrapy in sheep and scrapy in mice, curu in a child and curu in a chimpanzee inoculated with the child's brain tissue. Kreuzfeld Jacob from a brain biopsy in Oxford and the same process in the brain two years later of a chimpanzee inoculated with it. Next slide please. The same distribution, scrapy, natural and experimental, curu natural and experimental, Kreuzfeld natural and experimental demonstrating with a specific stain that shows astroglyos cells the fibrous astroglyosis which is a dominant part of the pathological picture and about as severe in this disease and this group of diseases as in any disease we know. On the other hand these are the only infections of the brain and I make no exceptions in infections. I mainly range through the whole range of such things as the protozoa through the viruses in which there is no perivascular cuffing, no leukocytic invasion of the brain parenchyma and no sign of a hypersensitivity or a inflammatory response. Next slide. For those clinically minded, next slide please. The phenomenon that these diseases proceed from beginning to fatal outcome with no change in spinal fluid protein and no pleocytosis at any stage of the disease is a further unique finding in fatal CNS infections. Now I'm not going to dwell on the hundreds of studies attempting to purify scrapy first using cesium chloride then using calcium and potassium tartrate, metrismid as well as sucrose and sucrose saline gradients have been used and the workers who have done this work are not workers who have difficulty using zonal ultrace centrifuges and purifying reverse transcriptases, viral subunits, P30 proteins, polyacrylamide gel, electrophoresing them but these same workers have been unable to purify by these techniques or any other electrophoretic focusing and the like the scrapy agent because no matter what small portion of maximum infectivity at a density of 1.2 to 1.25 they take, it spreads again and electron microscopic control of the re-zonal banding of it simply shows that we are trying to purify soap bubbles. The material obtained with enormously high infectivity is a three-layered bioelectron microscope plasma membrane light structure which fasciculates and the problem is that of concentrating and purifying membranes where the reactive receptor or the reactive macromolecule on the membrane is not known and although we assume it is a small nucleic acid all attempts with hot formaldehyde and with detergents to release this presumed genome have failed to obtain any infectious nucleic acid. Next slide. The most disturbing early data which came first from Compton working on scrapy then we reconfirmed it in using zonal banded and fluorocarbon cleaned brain suspended scrapy virus. We obtained linear inactivation curves at ergs 10 to the per square millimeter times 10 to the fourth. A energy input which if you are not familiar with radiation work would not lead you to realize that the most radio resistant forms of life we know to 254 nanometer radiation would intersect here at six. They would be the U1 variant of tobacco mosaic and bacillus radiodurans. This was interpreted first as absolute proof that the agents could not contain nucleic acid. In the suspensions if the argument is that they are dirty not sufficiently cleaned one puts a large variety of mammalian plant viruses and bacteriophage. These so called dirty suspensions have no effect on protection at this radiation. Next slide. I am not going to dwell on the long discussion this might entail but here is the same data you just saw but it simply shows that the plant extracts of the plant viroids these newly described smallest known viruses are even more radio resistant and the purified RNA genome of 115,000 molecular weight of potato spindle tuber virus is of the same order of magnitude of radio resistance whereas all conventional viruses fall here. Next slide please. This alone eliminates all other plant and animal viruses from consideration. Studies of very unsatisfactory zonal banding into 55 fractions in a sucrose saline gradient that the best anyone has obtained is a log scale so it doesn't plot out as well as the earlier ones I showed you. I have failed to reveal any relation with marker enzyme systems in enzyme activity for three lysosomal membrane bound enzymes and one mitochondrial enzyme. Next slide please. I am now going to rapidly summarize probably as fast as you can read. Much of what I have said simply to drive home the fact that these surely are an unconventional type of viral agent. On the physical chemical side I fail to mention what alone would leave any earlier microbiologist totally skeptical of the integrity of the workers. That a virus or any microbe can be stored in 4% formaldehyde or 10 or 20% formaldehyde at room temperature for years without losing tighter namely embalming fluid and that which we use in pathological laboratories to store specimens is blatantly ridiculous. It happens to be the fact. We have isolated Creutz-Feliaco virus from material that has been stored years at room temperature in pathological laboratories in formaldehyde. Scrapey and minkencephalopathy have been taken off paraffin embedded blocks which were for two years in formaldehyde before they were sectioned and left several years as blocks and obtained as viable microbes. Beta-propreolactone doesn't touch them. Ethylene, diamine, tetraacetic acid doesn't touch them. Proteases and the nucleases on even the deprotonized fluorocarbon treated zonal band infractions are not effective nor can we use hot formaldehyde to inactivate them and searching for double or single stranded RNA. Heat resistance is not incompatible with the biological experience but it is incompatible with most virological experience. Up until 80 degrees we have full stability. At 85 degrees we begin to inactivate and as with the hepatitis virus, boiling is not sufficient for hospital sterilization. Autoclaving however most certainly is. Next slide please. To go on further I have already summarized the UV resistance for you. I've mentioned the enormous ionizing radiation resistance. We use ultrasonic energy in such energy input that we would be inactivating rapidly polio or flu or most conventional viruses and all we do is gain in yield increasing our infectivity tighter. When we do an action spectrum at 2,370 angstroms the nanometer wavelength of 237 which is much less sensitive for inactivation work for most microbes, in fact all of them, we find six fold the sensitivity which is still not very high of that where it should be in a DNA RNA system. No one has visualized the agents and not one but literally dozens of the major structural electron microscopists working on the subunit structure of viruses have been involved for decades in the attempt and their starting material are either tissue slices containing an infectivity tighter of 10 to the ninth per gram or zonal banding material even more rich in infectious particles. That they contain no host proteins I mentioned earlier. We doubt this but it looks as though it may be the case and attempts to immunize animals with material that have infectivity titers as high as 10 to the 11th per gram have produced no neutralizing, no radioimmune assay obtainable and low cytotoxic antibody that we can with any of the existing systems that work with retroviruses and all other groups of viruses that we can identify. Next slide please. The pathological lesion is a coalescence of vacuoles which have been studied and studied and studied their membrane is not very nice to look at it doesn't look there's no visible very virus like structure on it. Next slide. It expands and blow next slide please it blows up the cell and finally we end up with neuronal destruction. A good deal of implanted electrode physiology done at Percy Camard and Marseille with animals inoculated having previously had electrodes implanted has yielded evidence that for literally months and sometimes years before neuron death, neuron involvement starts and by controlled perfused pathological study this can be verified. The first proof that we had an infectious agent here came when chimpanzees succumbed to the clinical disease Kuru. A disease which produced a clinical picture which no behavioral analysis analysts had ever seen in zoo or naturally observed chimpanzees or in laboratory subhuman primates. In fact all the experimental animals develop a disease which has never been produced artifactually which has never had a similar pathological or clinical entity develop spontaneously of unknown or known cause in laboratory animals of the species used. So there is really no control necessary. The next slide the pathology of these animals is identical to that in man with the complex perkinje cell and molecular loss in the cerebellum I'm not going to expand the pathological discussions. The chimpanzee interestingly enough develops a drooping lip months before our clinical neurologist and veterinarians are aware that the disease is there. Our keepers by personality change and facial expression are aware of the disease long before hard neurological signs appear. Months before. This is true of course with Kuru in New Guinea and where even child patients recognize their fatal disorder before any physician or their parents are aware of it. Next slide. The analysis of cinema at a stage too early for our neurological astute colleagues to detect the disease does reveal restricted forelimb movement patterns as opposed to the normal walking pattern which coincides with the drooping lip and the keepers note of personality changes. Next slide. This added matter is of incredible interest if one realizes that during the two and four years of silent incubation in cats and in many species of old world monkeys when these animals would be said to be normal and when their neuropathologically studied brain by conventional means would be said to be normal. We now know that there are vast disturbances from implanted electrode work in thalmic, hypothalmic cortical relations disturbing paradoxical sleep and other sleep rhythm patterns which Juve is so interested in and this is happening with all the viruses. In addition we are finding if we use perfusion and early sacrifice neuronal nuclear amyototic division by nucleate and trinucleate neurons, fusion of neuron to neuron and neuron to glial cell as we now know these agents cause in vitro as well in tissue cultures. And we have a whole gamut of exciting pathology going on in the normal animal years before disease only beginning to ask the question does this ever go on in man. Since it's associated with only what we would consider allowable personality disturbances and sleep disturbances and it requires perfusion studies to study it is a difficult question to decide upon and one many of us are not anxious to have investigated that thoroughly. Now the Kuru patient who first produced chimpanzee disease took 21 months incubation when that brain material was placed in other chimpanzees it dropped to 11 months by 13 months in fact many this has been done over many times and again we get a drop in incubation from the usual one and a half to two years to less than a year to 13 or 14 months on first passage. This immediate drop on second passage to a shorter incubation period has been done over again over 60 times. We know it occurs and with other species the same one passage adoption adaptation looks like a selection process which we are not familiar with in virology. We are more used to a step by step slow adaptation to a new host. It doesn't proceed further on further passage. By the oral route there was no infection in this animal 12 years later is still well having had millions of lethal doses of the agent by the use of skin liver a clean liver kidney of the serial suspension without brain we still got the disease and using peripheral non-intra cerebral inoculation we got the disease very early in the work. I'm not going into all the passage studies. Next slide. This simply shows the first demonstration that we could shift from the chimpanzee to South American monkeys, new world monkeys with the sacrifice of a short incubation period of one year and making our waiting time into two years again. This has not reduced on further passage. Next slide. The new world monkeys, a dozen species of them, owl, spider, squirrel are all now used in work with the human diseases and it has permitted the curtailment of work with the diminishing rare species of chimpanzee which incident in our breeding colonies we breed more rapidly than we use. We had 30 newborn chimpanzees last year and only use five or six. We're doing more. Eventually we may be able to return them to Africa where they're being killed more frequently and indiscriminately. The passage in chimpanzee is not continuing. We're using smaller animals and other hosts now that we can. Next slide please. I'm not going into the experiments that that involved. This is a biopsy of a European patient with Kreuz-Valiacov disease to reemphasize the punched out spongiform change in the cortex. Next slide. To show you that we also found chimpanzee passage in 13 months of this disease possible and then in 12 and 40 months in another chimpanzee but almost three years and almost four years when we went to capuchin monkeys and marmosets. In other words these smaller monkeys require much more patience in the experimenter. Again when we found we could go to small animals and finally cats. We see chimpanzee work but one line has been continued. Next slide. Now to depart I'm going to not use the small remaining time to elaborate extensively on all the animal work that has been done and continues but to tell you a little about the worldwide form of Kuru. One sixth of the cases are familial. These patients had the disease. They weren't autopsyed but it's a clinically recognizable syndrome. So here are two aunts and mother of the propositist plus his grandmother all dying of Kreuz-Valiacov disease. Next slide please. Another family from Germany with three generations of many proved autopsyed cases from which the virus has been isolated. In fact these genealogies which are every bit as good for a dominant inheritance when they are looked at in a whole as for Huntington's Korea for instance. Number over 60 now in the number of families we know of the disease and from 11 of the first 17 we have had brain biopsy material on. We have already isolated the virus and the remaining six are not negative. They have not been inoculated long enough. Next slide. So here we have a disease which would undoubtedly be acceptable as a gene control to retopharmidial disease where a degenerative pathology is virus determined and with no genetic control that we know of in the fully susceptible hosts. Here is a huge amyloid plaque staining TAS positive, metachromatic, agentophilic. The plaque that all of us have in our brains as we age some people unfortunately have a great deal of it before 60 years of age and we call it Alzheimer's disease. This plaque is present in 20% of the Kreuzfeld patients. It's present in 60% of the Kuru patients, even children. But since it is not present in the remaining 40% of the Kuru patients, brothers, aunts, uncles, and parents of the other victims who have it, we are sure it's a host phenomenon that has nothing to do directly with pathogenesis. On the other hand, in Alzheimer's disease it and the tangles are what we most focus on and it is very remarkable that a fair number of Kreuzfeld patients have the disease. Many lead to pathological disputes as whether they are Alzheimer's or Kreuzfeld until we finally isolate the virus. Next slide. Simply demonstrating the spherical symmetry with cross nickel prisms of the amyloid which by EM is amyloid and by long chain and short chain analysis is amyloid. Next slide. To bring you up to date on what is just in press a world epidemiology in an American of Kreuzfeld, we now know that anywhere we look hard we find.5 cases per million per annum death. This makes it a very rare cause of death indeed. But when one realizes that this means 200 deaths a year for the United States and we have not had 10 deaths a year for rabies with a 50 state surveillance in the last 20 years, it's fully 50 times more frequent than rabies probably 100. This is a minimum. We are actually finding one per million per annum anywhere in large cities we look intensively. The European studies now impress with Katala and Paul Brown working on them in Paris for all of France, parts of Germany, most of Italy and Spain are showing.5 per million per annum on first survey for the last 10 years with 1 to 2 per million per annum in some foci and large intense foci in Hungary, Slovakia, Palma in Italy and in Israel. Next slide. Kreuzfeld is a worldwide disease. The countries of Africa where it's been found are few and far between. Hungary has not been intensive. We don't know why the, we haven't been to Argentina, the other countries in South America, everyone we visited, we found it and in high incidence. It far outrivals, outranks rabies as a world cause of death from infectious brain disease by probably a factor of 100 on all continents. Next slide. The scrappy virus is probably also ubiquitous where sheep are kept, but most countries with it do not recognize it. We have, and British workers have carried it for decades through mice and to our horror after long incubation period of 73 months the cinemologous monkey developed a spongiform encephalopathy identical with Kreuzfeld's disease. Their scrappy sheep brain inoculation as did after almost three years squirrel monkeys. On passage the incubation period has dropped in the squirrel monkey, but not in the cinemologous. Here it has been passed back into sheep and goat with disease proving that it is still the scrappy virus biologically, although since no Kreuzfeld or Kuru virus go this way. We are very worried however since European strains of scrappy taken from the Compton laboratories once transmitted to monkeys lose their ability to go back to sheep and goat. Conceivably scrappy is the source of some of the human Kreuzfeld. Next slide. It incidentally can be found in the butcher shops throughout the world. There's a high selection for the butcher of all scrappy infected sheep and even laboratory infected sheep throughout Europe having passed decades ended up in the butcher shop. Scrappy is a worldwide disease. Next slide. Now in a rush travelogue I promised you I will try to tell you where this story began. It began not in this enormous second largest island in the world but in one small highland area here in New Guinea. Next slide. The Kuru region is at 2000 to 3000 meters with human population scattered in the eastern highlands of a very mountainous area where all the grass covered areas represent the effects of millennia more than centuries of human cultivation by slash burn agriculture. Next slide. The villages are small few humans see or saw in the stone age period when we first described Kuru more than a few hundred individuals between their birth and their death in adult life. The population unit is under 100 in most places up to 200 in some and never exceeding that. Next slide please. This is a village in which we're returning the area has rugged and the first year of patrolling throughout the area required of moderate amount of mountaineering. Next slide and bridge building. Next slide. But the next slide I'd go through these rapidly and the mesolithic level culture of the population involved with Kuru are people who quite significantly never washed in a lifetime covering their body with the rendered grease from their pigs which run wild and mate with the wild pigs or of their relatives. Next slide. The kunai fields are high grass which long carrier line is carrying our supplies through. Next slide. And in that area we would meet one of the 11 language groups which were defined during the Kuru research project since only two of them had had European contact at the beginning of our Kuru epidemiological work. This is an Anga Kuka group which is just on the border. Next slide. The villages contain groups of women who were slow to leave as most women on our arrival. Preferred not to walk but when tested could walk but with minimal ataxia. They told us through the one and two level child translation we had into a language we could finally use that they were dying of Kuru and they were invariably right. The disease rarely lasted more than a year and was characterized by midline cerebellary ataxia with preservation of cortical functions and no sensory disturbance. Next slide. The patients at this stage are unable to focus their eyes or hold their neck properly so she looks demented but with grunting signals can evidence very good contact with reality and good intellect still. However she is so ataxic that she can no longer stand without support. Next slide. And in every village of 100 people we found people dying with no exception. Next slide. We found over 300 deaths the first year and in a given 100 population village three women using sticks to keep ambulating to their gardens to bring food back to their houses were already evidence that they had Kuru. They agreed that they did have no foray woman required to stick to maintain her balance unless she had Kuru or some other injury. Next slide. The smallest patients were children of four or five years of age. Next slide. And the area we found was of enormous linguistic diversity. The Nugini people, the Melanesian speak over 1000 languages not dialects like French, Spanish and Italian or German and Dutch. But real languages at a dialectical language it's much more diverse than that. In the eastern highlands alone there is this enormous level of languages and at least five language families. Next slide. As we worked out the boundaries of Kuru, next slide, we also gave names to most of the people which as is always the case with indigenous populations they are the fallacious names usually an insult like the bastards over the hill used by their enemy neighbors. The people themselves have no names for themselves. So our fallacious names have stuck and the new generation children don't know that we are the source of them. And this is the boundary of Kuru against the language groups that had it showing that here culture and language have determined the boundary completely. And here culture and language have had nothing to do with the boundary of the disease. Next slide. When we counted up cases at the end of a year and this prevalence data can be read as yearly mortality the disease usually lasts about a year. So this was 1.6% of the population dying per annum. This went on for 50 years previously and has gone on through most of the first 10 years of surveillance. But there's been a change. Next slide. Please. This simply the Kuru area would be superimposed on here and this is decreasing population density into high altitude mountain ranges and virgin bush with no people. Showing that the people became less and less dense as we went in this direction. Next slide. But the running away from Kuru which they consciously attributed their location in the virgin forests too was with no success whatsoever since maximum incidents with 3% dying per annum occurred in the least dense most remote populations bordering on populations here that had no disease at all. But there's no intermarriage or contact culturally between these groups. There's no ecological shift. Next slide. We began to accumulate patients. They were willing to come in until we had from 40 to 100 at one time dying in the O'Kapa hospital which we built. Dr. Zegas, for next slide. A New Guinea government physician who discovered the disease and has worked on it with me since the beginning. Is Vincent Zegas here myself in an early day with a child patient at O'Kapa? Next slide. And we would have this boy himself knows he's dying of Kuru but hadn't convinced us yet his autopsy six months later certainly proved he was right. He is supporting the girl with the disease and these two have the disease and that young girl does too. And next slide. The moon fascist is a result of unsuccessful attempts at cortisone therapy. Next slide. Next slide. It simply caught a patient during the movement. I should be showing you cinema not still pictures. We don't have the apparatus. Next slide please. And a severe strabismus and extreme dysarthria. A final total neuromuscular incapacity with minimal loss of intellectual function characterized all patients. Children usually died in six months to nine months. Next slide. Fully a third of all the patients were children and most of the patients were female except among children where the male to female ratio became a little more reasonable. This 13 to 1 to 78 to 1 ratio of female to male in adult life was very difficult to imagine epidemiologically at first. Next slide. We eventually found that from genealogical studies that a one of three wives, the people are polygamous and this enormous death of women had reduced the sex ratio to three to one raised it in favor of males in the southern villages for polygamous society with a total sex ratio of three to one. This made it closer to five to one for marriageable age males. Polygamy on top of it made it very hard for many young men. The every case with no exception of the 4,000 we have studied has had similar intense family history. Next slide. I'm not going to dwell on the usual family history. At present we know from the dating of these histories that this patient here was the contaminating source for the whole crowd but this has taken years of epidemiological sleuthing. I'm not going into it. Next slide. And here is what's happened. An exotic disease that few of us have had a chance to see and now we tell you it is almost gone before you've got to see it. It has disappeared progressively and the male and female patients have left us with only a few deaths. This was plotted prematurely. The final number in 77 actually was under 20 for the total. Next slide. This year it will be under 20. Next slide. But the most dramatic thing is that these are taken at three-year intervals, leaving out the intervening year in case of epidemiological end of year prejudice. It doesn't make much difference. It's a matter of Christmas New Year, European physicians doing less patrolling at that time. It shows that the zero to nine-year-old patients rapidly decreased, decreased and then disappeared from the world. The early adolescent patients rapidly decreased but later, five years later, then disappeared. Finally the older adolescent patients decreased and then disappeared. And now the 20 to 25-year-olds are gone. Every year the youngest patient is somewhat older. The law that no patient who was born since Kuru cannibalism stopped in his village has ever died of the disease holds. It tells us a great deal more than that first remark might immediately reveal. It tells us that the suspected transplacental transmission, milk factor transmission, vertical transmission from mother to child born during the disease or born before the mother is ill is not occurring. We're it occurring, this incredibly uniform and if I projected to 78, still more uniform with this mostly gone and that gone, disappearance would be unaccountable. Next slide. We do not have any such knowledge for the familial cases of Kreuzfelljacob in Germany and the rest of the world. If one doubts our ability to assess age of these now disappeared types of patients, pre-pubertyl children of both sexes, here is a group on dimer caprol and calcium-versonate therapy when we were looking for a Wilson's disease like copper or molybdenum toxicity and failed to find it. We had many false alarms in the beginning and all of them died of Kuru. There are no such patients in the last 10 years in the world. Next slide. The Kuru epidemiology was done mostly by the Robert Browning technique of the Pied Piper of Hamlet with assistance of here who are all married polygamous men today. Next slide, they rapidly, a group of some 30 of them provided, next slide provided us with translation of all 11 languages into the one language we learned. The young men who were leaving the area had to be initiated into adult life at which stage of life, all still pre-pubertyl, they have totally left the women's society never in their life even as polygamous adults to enter the house of a woman in traditional culture. All copulation and procreation was a daylight affair in the gardens where the privacy was greater than that obtained in the most remote Swiss valley and the phenomenon of sleeping with a wife is unknown to Highland New Guineaers until missionaries introduced this strange custom to them and the phenomenon of crawling in darkness for copulation under a roof is a phenomenon unheard of and rather repelling to them a subject of jest not to be taken seriously. Anointed with pig grease and never washed this would be human grease in a cannibal ceremony next slide but only for the smaller children and never the males. At this age people have left the women's culture only the women opened the Kuru victims bodies and contaminated themselves and children with them. Next slide they celebrated their initiation into manhood. Those three young men incidentally are all college graduates in the United States today. Next slide they are I'm not exaggerating. Next slide but this is the age at which all warfare took place most New Guinea Eastern Highland groups left that for the adolescent sex segregated culture while the old men as in most cultures planned the wars but didn't take part in them. Next slide and this age produced the sex segregation which did produce this enormous difference in sex predominance of the disease. Next slide girls at a early pubertal age were married with pig omentum on their head. Next slide and usually the young wife of a polygamous older man older widows went to the adolescents boys as their first wife. Next slide and all cooking was done over steam cooking with no vessels in pits of heated rock a meter and a half under this pile of several hundred pounds of pig meat and vegetables. It's an autoclave essentially a volcano like peep is earth is heaped on top of it and there is very little enteric disease since it comes right out of steam cooking. So the temperature attained would have been insufficient to inactivate the Kuru virus and human tissue was similarly handled. Next slide on the other hand with men handling pig tissue this would have been women for human tissue and the children around would have been infected. We believe the all infection was through mucus membranes and skin not orally it takes multi million lethal doses to infect with scraping orally only a millionth as much through skin or mucus membranes and with current cranes felt it doesn't take orally it is not eating the dead even though Claude Levy Strauss would have it totem wise symbolically that way. Next slide simply showing the children's involvement with the mumu pit of steam cooking. Next slide and finally the disappearance from the world of childhood Kuru ten years later the disappearance from the world of adolescent Kuru now the disappearance of young adult Kuru and hopefully within 20 years the disappearance from the world of Kuru how we will get rid of Kreuzfeld Jakob disease is another matter. We can tell you that only a few percent of the cases in the world five or six percent are attributable to the atrogenic cause. In Suric the use of stereotactic electrodes contaminated from a Kreuzfeld patient has produced two deaths years later in young people who were cured of their epilepsy by the surgery only to die from Kreuzfeld Jakob disease. Our form of neocannibalism brain use of in brain surgery of durameter from older patients and the use of corneal transplants from victims of Kreuzfeld Jakob disease has in fact caused the disease. Most major American and European clinics if thoroughly studied and if they have good enough records can detect the case caused by their neurosurgery. We do have a series of several dozen atrogenic cases the complex medical legal aspects of the issue and the humane aspects of the panic produced by epidemiological questioning of those in danger has restricted reporting of many of them. On the other hand we do already have the published accounts of a half dozen. One of the easiest ways of course to our utter horror of finding the virus is visiting a neuropsychiatric clinic where it would accumulate and we are just now aware of this hazard and changing the sterilization methods necessary to prevent the atrogenic spread since conventional sterilization with alcohol, ethylene oxide, sterilizers, zephyrin and many of these usual hospital antiseptics don't work. Because it smelled like hospital should in two, 30 years ago with Clorox, permanganate and iodine around which none of us and as young physicians want to take home to our families any longer on our clothes did inactivate the agents phenol did but the modern most of the modern antiseptics don't. There has been warning in all the world literature especially in the neuro and to pathologists neurosurgeons have died of the disease pathologists and dentists have we do not know and have no way of knowing whether the number that has died is a significant increase above what would be expected because of course they are closer to the medical profession and this rarely made difficult diagnosis is really even now missed in major academic neurological clinics as we are discovering in the worldwide epidemiology. I leave you then with a failure to have told you about the plural, the phenomenon that the Kuru focus is not the only intense focus of neurological or other chronic disease in primitive populations which we have pursued and are we are pursuing others, amyotrophic lateral sclerosis foci of 1000 to 2000 fold the intensity of any other civilized part of the world in West New Guinea populations and keep an ince of Japan and Guamani and Micronesian populations. Obviously if we ever find the answer to amyotrophic lateral sclerosis in these small enclaves which can be visited in a week where this has become the first cause of death in adult life are the place to find it. Thank you. Thank you.
at times several decades after original infection and a long, silent incubation period. The second discovery was that of a new group of and biochemical, and biological properties that central highlands of eastern New Guinea who their dead kinsmen, they contaminated the skin them. This led to a fatal brain disease appearing surveillance of all the population involved for caused by a virus indistinguishable from that of means of transmission of our civilized form of mink encephalopathy of mink. We call the group hypersensitivity types of immune response, B cell non-host protein in their structure. Instead, replicating membranes devoid of nucleotide to establish unequivocally that a nucleic acid is like agents. We are dealing with a new group of disease of sheep and goats throughout the world spongiosis and although it was previously thought process in the brain two years later of a in this disease and this group of diseases as in the brain parenchyma and no sign of a tartrate, metrismid as well as sucrose and conventional viruses fall here. Next slide laboratories in formaldehyde. Scrapey and embedded blocks which were for two years in and as with the hepatitis virus, boiling is not material even more rich in infectious particles. first proof that we had an infectious agent here came when chimpanzees succumbed to the clinical spontaneously of unknown or known cause in conventional means would be said to be normal. We doses of the agent by the use of skin liver a a short incubation period of one year and making These patients had the disease. They weren't children. But since it is not present in the the virus. Next slide. Simply demonstrating the world cause of death from infectious brain rugged and the first year of patrolling slow to leave as most women on our arrival. bush with no people. Showing that the people neuromuscular incapacity with minimal loss of intellectual function characterized all patients. Children usually died in six months to nine patients were children and most of the patients contaminated themselves and children with them.