University Place

Science and Art of Prions and Other Pathogens

– Welcome, everyone, to the special Thursday night edition of Wednesday Nite at the Lab, I’m Tom Zinnen, I work here at the UW-Madison Biotechnology Center. I also work for UW-Extension, Cooperative Extension. And on behalf of those folks and other co-organizers, Wisconsin Public Television, Wisconsin Alumni Association, and UW-Madison Science Alliance, thanks again for coming. Tonight, it’s a special Thursday night edition. Usually I say we do Wednesday Nite at the Lab every Wednesday night, 50 times a year. This is a special occasion because we’ve received a special gift from an alum, Pamela Caughey, who is an artist. She has given us an extraordinary work of art called Ubiquitous, Migration of Pathogens. It’s inspired in part by some of the work that her husband, Byron Caughey, has done. And they’re both here to talk with us about their respective works. Pamela was born in Milwaukee, Wisconsin and went to high school at Grafton. Then she came here to UW-Madison and got an undergraduate degree in biochemistry and then received an MFA at the University of Montana. She is now a full-time exhibiting artist and she teaches workshops around the world. She’ll be speaking second tonight.

First, the lead-off batter is Byron Caughey. He was born in Peterborough, New Hampshire and went to West Town high school in Pennsylvania. Then he went to his undergraduate degree in Colorado State University at Fort Collins and studied chemistry. He came here to UW-Madison to get his PhD in biochemistry, did a postdoc at Duke University, and then took a permanent position at the NIH Rocky Mountain labs in Hamilton, Montana. I’m delighted to be able to welcome you all here. Very much appreciate the gift that you’ve given us. It’s on permanent exhibit now here in the atrium of the Biotech center. First up is Byron who’s gonna be talking with us about his research in prions. Would you please join me in welcoming Byron Caughey to Wednesday Nite at the Lab. [audience applauds]

– Can you hear me all right? Well thanks, Tom, for the opportunity to talk about my work. I’m really just the warm-up act for Pam, who’s going to give you her artistic impressions of this new world of pathogens. But I’m going to focus on an aspect of, to use a broad use of the word, microbiology, and that is, how corrupted proteins can act as pathogens. Normally you think of pathogens as being viruses, bacteria, fungi, protozoa and so forth and so on, and all those pathogens have proteins for sure. But they also carry their own genes around with them, and other molecules as well, from host to host, to enable them to replicate. But what’s more recently appreciated, if appreciated is the right word here, is that proteins that are made by our own bodies can also be corrupted into forms that have a self-replicating activity that, at least, can allow, even in the case of Alzheimer’s, Parkinson’s and related diseases, in the spreading of this corrupted misfolded and pathogenic form of our own body’s proteins. And sometimes it even can lead to them being transmissible between individuals as sort of infectious agents. That’s sort of the bad news.

Part of the good news I’m gonna tell you about is that we and some other labs have exploited this self-propagating or self-replicating activity that these corrupted proteins have in order to improve the diagnosis of these diseases, which can be very hard. All of these many neurodegenerative diseases can be very hard to clearly diagnose, especially early in the clinical phase of disease. So it’s important to have accurate diagnostic tests. And we also are targeting the self-replicating activity, at least one way or another, in certain treatment strategies. This is Pam’s rendition of a lump of this sort of pathogenic, infectious form of protein that we often call amyloid plaques. But just to back up and give you a little perspective into– Let’s see what– Oh. A little context as to where these plaques occur. Let me start by showing you a picture of the brain, obviously, which of course is incredibly complex organ that contains actually 100 billion cells or so. Very complicated. And each one of these cells contains one or more billion proteins.

In fact, of those billion or so proteins in each cell, there might be 1,000s or 10s of 1,000s of different proteins in each cell. So it’s a very complicated business here. And of course given that fact, relative to the size of a cell, this would be a neuron or a nerve cell and signals coursing through its veins, so to speak, proteins are very tiny compared to the size of a cell. As you may know, proteins are linear chains of amino acids that provide us with enzymes that are the chemical factories of our bodies that carry out chemical reactions and basic metabolism. They also provide structural and mechanical elements of organisms, including our cells. They regulate, mediate most physiological processes. And prion protein, or we’ll call it PrP, is expressed normally in all of us and all mammals. This is just a picture, cartoon, of this string of amino acids that make it up. But, as you may well know, for proteins to function properly, they have to fold up properly into very well-defined three-dimensional structures to be active, but along the way, or even after this folding process has occurred, problems can occur that allow the exposure of sticky surfaces on these protein molecules that cause them to clump together. They can then reorganize themselves into highly ordered complexes or aggregates of these proteins.

And oftentimes they can, under certain conditions anyway, can form these ribbon-like structures that can clump, can grow on the ends and clump together into large clusters of what we would call fibrils or filaments, amyloid filaments, in the body. Now, normally we have pretty effective protein quality control mechanisms that keep this sort of– these misfolded forms of these proteins, at bay. But sometimes things can ravel out of control, allowing these misfolded protein aggregates to grow and grow and grow and accumulate, especially in the brains to the point where they cause deadly diseases. Now, I don’t know if you can see it right here, but these are slices from mice brain slices, taken from mice that were infected with a tiny pinprick’s worth of a misfolded protein called a scrapie prion about 40 days prior to this first picture taken. And then right at the beginning, they’re so little there you couldn’t even see it. But we’re able to stain this abnormal protein in the brain, starting at about 40 days after that initial infection. Then it, I think it says 80 days, you start to see a little more, whatever that says, 180 days, you see more and more and more until it finally starts to fill up the brain to the point where it really causes so many problems. It’s directly toxic to nerve cells but also it’s just preventing proper interactions between the cells of the brain, and the animal is then euthanized. So an individual lump of this material here that can accumulate in the brain is called an amyloid plaque, as you can show here in a much higher power image. A lot of different what I showed you here is, are aggregates composed of this prion protein or PrP when it’s corrupted into its abnormal and infectious form.

But there are a bunch of other proteins in humans alone that can misfold similarly oftentimes and cause terrible diseases. Let me start with prion protein or PrP itself, can give rise, it’s the misfolded protein that’s responsible for chronic wasting disease in the deer and elk here in Wisconsin and all over North America. But it also in humans causes Creutzfeldt-Jakob disease, which is a devastating, very rapid and deadly disease. There is a protein called beta-amyloid that’s partly responsible for Alzheimer’s disease along with misfolding of a protein called tau. But tau alone can cause a bunch of misfolding, and accumulation of that can cause a bunch of other brain diseases, dementias and so forth. Alpha-synuclein is yet another protein whose misfolding is responsible primarily for Parkinson’s disease, dementia with Lewy bodies, multiple system atrophy and some others, and so forth and so on, including Huntington, which is a protein that misfolds to give rise to Huntington’s disease, which is actually a genetic disease. A lot of the time, these proteins can misfold sort of spontaneously. You don’t have to have a mutant form of it to do the dirty work. But in the case of Huntington’s disease, it’s usually a mutation of Huntington that causes this. Of course collectively, you’ve heard of a lot of these diseases.

And they represent a huge disease and economic burden on society. Doctor Johnson was telling me earlier today that they estimate that Alzheimer’s disease, I think alone, will cost society about a trillion dollars a year by 2050, as we are all getting older and these diseases become more prevalent. So a huge problem, not to mention the problem for the patients themselves and their families. And most of these, as I’ve mentioned, have the fibrillar or filamentous amyloid deposits that can propagate within given hosts, a person, as if they’re replicating prions or infectious agents. Several of these important diseases, namely Alzheimer’s and the tauopathy, the other tauopathies listed here and synucleinopathies like Parkinson’s have been shown to be experimentally transmissible from patients who have these diseases into laboratory animals that have been engineered to be susceptible to these diseases. So it’s been shown in the lab that you can take a little bit of tissue from a patient, put it, say, a small amount of it into the brain of a susceptible rodent model, so to speak, and have that material expand, propagate and start to cause the pathologies that’s similar to the diseases that you see in humans. The real question before us now, we really don’t have clear answers in most cases to this question, is, okay, you can do this by forcing the issue in experimental animals. You take a lot of tissue from a human and you jam a lot of it into their brain and yes, bad things start to happen. But the real question that’s before us is whether these diseases are actually transmissible from one person to another in the real world. I don’t mean to suggest for a second here that anyone that I know of thinks that these diseases are in any way casually contagious from one person to another.

What we’re thinking about is the possibility primarily of the disease being transmitted through invasive or neural invasive medical procedures, for instance, like neurosurgeries or transplants or transfusions, that sort of thing, where you’re really forcing the issue in patients as well. That is due to medical procedures. So the key question is not just, are these fibrillar aggregates able, you know, are they able to propagate wildly in a test tube or even in a laboratory animal, but are they, if you put them into another host, are they infectious or noninfectious? In other words, can they really replicate, say, in a person who receives some of this, replicate enough to the point where they actually cause disease, that is, they’re pathogenic as opposed to nonpathogenic? So these are really important unanswered questions that we have to deal with as we go ahead. And again, just because you have one of these amyloid fibrillar forms or amyloid fibrils of a given protein, it may be highly infectious and pathogenic, like this stuff here, which is a pure P-based scrapie prion shown in electron micrograph here. In fact, this stuff is so nasty that, if you take, say, a milligram or, say, roughly a pinhead-sized little lump of this stuff and dilute it into liquid and were able to inject that material into hamsters, it could cause a million or a billion hamsters to die, just that one little bit itself. So incredibly infectious and pathogenic. So pathogenic, in other words, causing disease problems in the brain that it causes clinical disease and death in these animals. On the other end of the spectrum, we and many other labs have made perfectly good self-replicating synthetic amyloid fibrils of the same protein, PrP from hamsters, that actually is totally not infectious nor pathogenic, nor causing a clinical disease. You could put a whole pinhead’s worth of that protein into the brain of one animal and it would not cause a problem. So there’s an incredible spectrum of possibilities here just with one protein alone, that is hamster PrP.

Again, these are composed of the same protein sequence but we know now, using various biochemical measures and so forth, that they’re differently folded and assembled in subtly different ways that you can’t really see in these electron micrographs shown here. And even though all can grow continuously, endlessly, in test tube reactions, they range from being highly deadly to totally innocuous if you inject ’em into the brains of hamsters. Bearing that in mind, I just want to give you an example of what we think the worst-case scenario that we know of in nature in terms of one of these infectious pathogenic prions, and that is the example of chronic wasting disease in deer and elk, which of course you know it’s here in Wisconsin, it’s spreading readily, I think in at least 20 other states in the US and in Canada and Korea and now in northern Europe. Actually the whole transmission cycle in these deer or elk or moose or whatever is so efficient that it can contribute to local prevalences of 30%, to even 50%, even in free-ranging deer. Often you find it in game farms, but it can be in free-ranging deer and is to such high numbers now that it’s actually affecting, dropping, the overall population levels in deer populations, in North America at least. And nobody really knows what to do about it or how to stop it. It’s really a kind of a sketchy thing. But the process that’s so efficient is that somehow these animals can take up the prion, this infectious chronic wasting disease prion, into their bodies, it replicates, it moves from whatever peripheral site of exposure that they had into the central nervous system to the point where it kills the animals. And all the while that they’re infected, it’s shedding large amounts of this same stuff back into the environment where it can last for years and years. And then in a form that can be taken back up by other animals.

So this is a very efficient but complicated process. As I mentioned, you can see here’s a map of incidence throughout North America, with a nice big cluster here in both free-ranging animals shown in gray here, but also yellow and red showing game farms, where it had either been depopulated or still there. So the big question that we have in our heads and we don’t really know the answers to much at all yet is how transmissible chronic wasting disease might be to people or animals, livestock or other species, that are exposed to these infectious agents. But it’s really important question going forward. Then we know that a number of these other aggregated proteins can spread in a prion-like manner within the body of people to cause these various diseases over here, at least some of them. But we still really don’t know the question and it’s really important to get to the answer of this question, whether they’re transmissible between individuals in real life, under any real-life scenarios that we might encounter. And this is a little bit difficult question to ask compared to, say, a discussion about other types of pathogens because compared to conventional pathogens, these prion-like agents or prions themselves elicit little classical immune response or antibody response in the host because they’re actually made of host proteins. So they’re just differently shaped. That can sometimes lead to an antibody response, but it’s very little compared to what you usually get with other types of pathogens. And again, antibodies are often used to detect infections by other pathogens, whether they’re viruses or bacteria or whatever.

And they also carry no genes of their own, these protein prions. And genes can often be used to highly sensitively detect infections with other types of pathogens. So yeah, they’re very hard to detect. And the other problem is that they may not be decontaminated by typical cleaning procedures that are used in clinical or agricultural settings. So this is a difficult question to answer. I would say right at the outset, there are no signs of big problems. You think if, you know, Alzheimer’s or one of these incredible, Parkinson’s, incredibly common diseases were actually at a large scale being transmitted between people, we’d see, it’d be loud and clear. But even if it’s happening on the margins, you know, only occasionally, when you have the disease as devastating and common as those, some of these diseases here, you really need to know about it. And you want to avoid it. So despite there being no big problems, there are hints of transmission of some of these important diseases that are emerging in the literature.

And just to give one example of multiple ones that talk about this amyloid-beta pathology, so amyloid-beta is this protein that accumulates into these pathological clumps in the brains of Alzheimer’s patients, for instance. So this is a slice of Alzheimer’s brain and here’s the plaque. And there are a couple or several papers now, recent papers, providing epidemiological suggestions of transmission of this A-beta pathology under some circumstances between people. And one of those scenarios, or one of those observations, was that all young-onset cerebral amyloid angiopathy patients, it’s a difficult, a little bit different type of disease of people that involves this amyloid-beta pathology around blood vessels, were found to have undergone neurosurgeries when young about 30 years earlier, raising these authors here to suggest that maybe this was actually due to contamination of the neurosurgical instruments that were used on them with this beta-amyloid material that was then able to propagate in their brains and cause them this horrible disease. Also they’ve seen this amyloid-beta pathology in young patients who received injections of growth hormone that had been contaminated, or had been isolated from people who had died, from cadaveric pituitary gland. But it was then later shown to be contaminated with A-amyloid-beta and also a tau protein that’s involved in Alzheimer’s pathology, so again suggesting that maybe that had something to do with them getting this pathology. And then also, the same sort of thing’s been observed with patients who have received dura mater grafts, that’s the tough sort of membrane that is sometimes transplanted from one person, from a cadaver, to a person getting neurosurgery as part of the surgical process, and it’s supposed that maybe those grafts were contaminated with this amyloid-beta pathology. So really there’s multiple studies now showing this sort of thing. I think at this point they provide a correlation between the presence of amyloid-beta pathology and these sort of procedures, but don’t yet, at least as far as I know, necessarily establish a cause and effect relationship there. So what we’re left is wondering, are these just rare circumstances where this sort of thing can happen, or are these tip-of-the-iceberg suggestions of much broader risk of this kind of transmission process? But, you know, when wondering about this, it is worth noting that each of these same roots of transmission have been documented for, in the case of human Creutzfeldt-Jakob disease, again where full-blown disease was really known to be transmitted by these kinds of mechanisms.

So it’s not unprecedented, precedented. So it leaves us with a big, gaping question of, which of these, if any, might be transmissible between people, if nothing else, by rather invasive neurosurgical or other types of medical procedures? Again, not casually by contact in the street or whatever. So there are warning signs, but a lot more information is needed to get to the bottom of it. Now I’d like to turn to this issue of using this self-multiplying activity that these protein aggregates have to help in the diagnosis of these diseases. The big problem in dealing with all of these different neurodegenerative proteins, this folding diseases, is that early diagnosis, at least clear early diagnosis, can be very difficult. And the final diagnosis, in fact, often depends on identification of these misfolded proteins when you examine the brain after the patient has died, which of course is too little, too late by a long shot. So our goal, and the goal of a number of other laboratories around the world, is to improve this early diagnosis of all of these terrible diseases by exploiting the amplification power that’s inherent in their pre-amyloid-seeded growth mechanism to develop ultrasensitive tests that we can apply to diagnostic specimens that we can get from patients while they’re still alive, and yet get it easily. So what are we trying to do here? Well, basically we’re exploiting this growth cycle that I’ve been talking about. You have these clusters of, these abnormal clusters of a given protein that we would call a prion seed here that are capable of grabbing on to the normal form of the protein, given protein, that’s usually floating around by itself and doing its job. It can grab onto it, change its shape, and in doing so, grow through successive iterations ’til you get a longer prion seed or prion fibril and you started with it, if you fragmented, you’d get more prions than you had to start with.

It’s as simple as that. And so once this happens, if you introduce this seed or seeds into someone or an animal, this can go round and round and round this cycle and just amplify by many orders of magnitude to the point where it causes disease. So how have we exploited this in a practical sense? Well, we figured out, and this is work primarily of postdocs, former postdocs of mine and a current postdoc of mine. What we do is, we take a plastic plate that has 95 little wells in it and we mix together a test sample, some sort of fluid or tissue specimen from a patient, together with a vast excess, and this test sample may or may not have these seeds. But we incubate in a vast excess of the normal protein molecules together with a dye, a fluorescent dye, that’s sensitive to the presence of the amyloid form, that’s these long aggregates and structures here, and then fluoresces dramatically in the presence of amyloids. So we put this plate into a machine that shakes it and incubates it and reads fluorescence over the course of time. And if you have a lot of these prion seeds right here in your sample, you can get very rapid, massive increases in fluorescence in your test reactions. In fact, these are extraordinarily sensitive because we can, what we’ve done in this experiment is put into these wells a little bit of scrapie brain, like, very tiny, tiny volume of a homogenate of a brain from a scrapie-infected hamster. But it’s only out to the point where you have diluted that brain to a billion-fold that you start to lose detection of the prions in that sample. So extraordinary amplification here.

Whereas you get no signal if you put a massive amount or any amount of normal brain tissue in that sample, as you can see here. So we’ve designed these tests to be extremely sensitive. We get billions or trillion-folds amplification of these seeds here, right there in the test tube, to the point where we couldn’t see it to start with but we can sure see it now with this fluorescence now. And in fact, in the case of human variant CJ brain and sporadic CJD, for that matter, two different types of human prion disease, we can dilute their brain tissue sample 10 to the 14th-fold. I mean, it’s just an enormous, it’s– What is that? 500 billion-fold or something. I mean, it’s a massive number, whatever that 10 to the 14th is. And still pick up the presence of the prions. They can be quantitative, so you can use these assays to measure not only if the prions are there but how much. That’s really important in many applications. And we design them to be really disease-specific in awful lot of ways.

So we’ve applied this strategy to, not only to prion diseases, the ones that are based on this pure P-protein, but also these other types of protein, or some of these other protein misfolding diseases. Let me just give you a quick synopsis in terms of each of these types of diseases. We call these assays RT-QuIC assays. We don’t need get into why. But for these pure P-based prion diseases, we’ve got them now. We and other labs have developed them for virtually all known prions of mammals, including humans. And most importantly for humans, we now can use these for the first time to get a 100% accurate diagnosis while the patient is still alive when it comes to sporadic Creutzfeldt-Jakob disease, which is the most common form of human prion disease, and it’s a terrible, terrible disease, if you can use samples of their spinal fluid, and if need be, nasal brushings. And we’ve also shown recently and published in the case of the skin, that skin and eye-based fluids or tissue specimens should also be useful, some of this being unpublished. But this just shows the kind of fluorescence reaction we get from nasal brushings that we’ve taken from CJD patients, cerebral spinal fluid or spinal fluid samples shown here. And this is the non-prion disease control samples down here.

So we get very good performance out of these prion assays. We’ve also developed ’em for these various diseases, including Alzheimer’s and PSP, CBD and Pick’s disease that are called tauopathies because they involve the misfolding and accumulation of a protein called Tau. So we have distinct assays. We haven’t published all of these that have been optimized for these different types of tauopathies. Again, these are ultrasensitive. Can detect down to billion-fold dilutions of brain specimens. And we see incredible selectivity for the given type of tauopathy for which the given assay was optimized. And this sort of selectivity in their growth properties or replication properties might explain why we have different types of tauopathies made from the same, very similar protein sequences, such as these shown here. So we’re very encouraged about this for the tauopathies. And then we also have, we and two other labs have developed very nice assays for synucleinopathies.

Those are including Parkinson’s disease and dementia with Lewy bodies that involve the misfolding of this protein called alpha-synuclein using very small volumes of spinal fluid taken from these patients early on in their clinical course, before they’ve even been treated with L-dopa to try to figure out if they really have Parkinson’s disease or not. As a rule, we pick up the vast majority of patients, we get positive signals above this dotted line for almost all Parkinson’s and Lewy body dementia patients, as you can see here, which is very helpful, we think. So overall, these sorts of assays, we think, have the great potential to provide more accurate and early diagnoses of these diseases. And drug companies are contacting us all over the place because they’re excited about the prospect that these tests can improve the patient selection or cohort selection for their therapeutic trials as they try to develop drugs for these diseases, because it can be a really sketchy proposition, especially early on in disease when they’d like to begin their clinical trials. You need to know the people you’re treating have the disease that you’re trying to treat, after all. So they’re really excited about that and the possibility of being able to monitor the amounts of the pathological protein aggregates in these patients over time, with a marrow treatment. So overall, just to sort of summarize, we have fantasized a little bit, we think that we should be able to develop a large panel of these RT-QuIC-like amplification assays to enable the early and accurate diagnoses of a whole bunch of, most if not all, of these diseases, which would be very helpful, using a variety of tissue specimens that should be fairly readily available. Again, to improve prospects for treatments and therapeutic trials, because of course the earlier you can clearly diagnose a disease in a person, the easier it is to treat it. Finally, I’d just like to briefly mention our efforts– Well, strategies, first of all, that various outfits are taking to treat these kind of protein misfolding diseases. Companies and institutions of all sorts, researchers are trying to develop drugs that block this misfolding process that then happens here.

They’re trying to promote the immune system’s attack on prions or prion-like aggregates that may be accumulating in the body through vaccination approaches or just straight-up direct passive injections of antibodies into people. A lot of companies are working on this. There are also efforts to improve the protein quality-control mechanisms that seem to break down as we age. And if you could try to jumpstart those again so they can clear these back out of, especially the brain, because the brain doesn’t turn over much at all, then it might be a good strategy to try to do that. And a strategy that we’ve taken that we’re kind of excited about is simply one to reduce the supply of the normal protein here that needs to be there in order for this pathological dastardly aggregate to grow. There’s a couple approaches you can take. One approach that we’ve taken is to knock out the messenger RNA that encodes this protein. In order to make the normal protein in the first place, the gene in your chromosome has to be transcribed. The code has to be read and converted into a messenger RNA molecule that then provides the code for translation of the normal protein. So if you knock this messenger RNA out, then you knock out the ability of the body to make as much of the normal protein.

And so simply, we have, I mean, much more simply than it actually is, we use a kind of molecule called an antisense oligonucleotide or an ASO that can affect messenger RNAs for individual proteins. I mean, you can target the messenger RNAs for a specific protein, not all the other ones that are in the body, the one that you think is causing problems. And this sort of acts like a molecular Velcro that’s sequence-specific. The small oligonucleotide that can latch onto the messenger RNA that provides the code for the protein you’re trying to knock down and either block the translation or cause the messenger RNA to be destroyed, reducing the production of this protein. Also there are ways to use these ASO molecules to modulate the translation of this protein to give you an altered sequence of the protein rather than knocking it out. Just to finally– There are now, one thing that encourages us is that there are now ASO treatments in development or even in the clinic available for several neurological diseases. What happens with these compounds is that you can now deliver them to the patient by injecting them into the base of the spine, into the intrathecal space, once every few months or so. And it’s like getting a spinal tap or lumbar puncture. And they have ASOs now for this horrible disease called spinal muscular atrophy. This particular ASO has now been approved by the FDA for years.

There’s one against Huntington’s disease that knocks down, it’s not in the clinic yet but it’s been shown to knock down the Huntington protein that causes this disease, and at this point in time is known to be well tolerated in humans and actually reduce the levels in the spinal fluid. There are ASOs against another protein that causes ALS, Lou Gehrig’s disease, and also the tau of Alzheimer’s disease, that are under development. And in terms of prion disease, we’ve been collaborating with scientists at the Broad Institute and Ionis Pharmaceuticals to try out anti-PrP ASOs. And in our studies we found that simple injections, or even sometimes a single injection of an ASO into the brain of the mouse can double their survival times, which is saying a lot because these are long, drawn-out diseases. So there is hope. I’m not, I just wanna end by thanking you for your attention and thanking all the people, and many, many people that are critically important in the work that we’ve been doing towards the ends that I’ve described to you here. It involves a massive effort by many clinical centers around the world to obtain the necessary diagnostic specimens and clinical expertise and biochemical expertise and everything else under the sun to get this sort of thing to work. So, thank you. [audience applauds]

– Okay, there we go. All right, well thank you all for being here. I really appreciate it, especially my artist friends who are here. [chuckles] I’m not used to speaking as a scientist, so it’s really great to have a few [chuckles] of my like-minded people here. Not that you’re all not like-minded, but anyways. So I am here to, number one, thank both Daniel Einstein and Thomas Zinnen and all the people that worked with them to make this possible. It’s quite a large installation, and walls needed to be reconfigured and reinforced to hold the heavy glass panels. So far, nothing’s broken, so [chuckles] keep our fingers crossed. The purpose of this, for me to be here, number one, I feel really, really just thankful that this will be on permanent display here at Biotech and Genetics. I’m completely honored, and it’s been wonderful just walking around campus today. I haven’t been here in over 30 years. To really walk the campus, it’s been really an honor.

So, “Ubiquitous: Migration of Pathogens” and how it came to be. That’s kind of a long story. But this, before we get into that, this is an overview of just a little section of campus that I became very familiar with. You’ll notice that the lower arrow is pointing to Biochemistry and is pointing to the backside of Biochemistry. That happens to be where I had a conversation with Byron. I won’t go into what we talked about, because he asked me not to tell you. [audience chuckles] But suffice it to say [laughs] that it was one of those getting-to-know-you conversations and it probably, the two of us probably found our way up to this top arrow at some point. This is on Linden Drive. If anybody is here, they know what that building is. I just thought I would throw that up there because we all know what they make.

It’s Babcock Hall. Thank you, food scientists. We’re really, really thankful for what you do here. I might have been in the wrong field, because I think I would have done really well in that particular field. Anyways, we all have our favorite flavors. A brief history about me and how I got from being in the School of Biochemistry to becoming an artist. Now you have this Ubiquitous thing, and I have to say it started here. So clearly, that installation would not be there if I didn’t come here first. I attended the UW-Madison campus from 1980 to -83, a long, long time ago. And I don’t know, second year or so, I met Byron. He was a graduate student, I was an undergraduate. You know, I was merely going along with my coursework and got to know him. I just kinda started to see that my brain didn’t quite work like his. He’s logical, he’s analytical, he’s practical. And I started to realize, “You know, I’m really not sure I’m in the right field.” [audience laughs] Anyways, I stuck with it, got my degree. We eventually got married. And then we found ourselves moving to London for a year, which I’ll talk about later. And then after that, we went to Durham, North Carolina where Byron did his first postdoc. And then we ended up in Montana, where we live now.

We’ve lived there for 31 years. Between 1986 and 2006, Byron encouraged me to pursue what I loved, and that happened to be art. He said, “You know, you don’t have to go work in a lab. You can pursue art.” We had a newborn, our oldest son, Kalen. During that time, the next 20 years, art really became my science. And I do think that science is an art and art is a science. I think having been exposed to both, I feel like they kind of are really combined. The definition of science being that, something that can be studied. Well, we can study a lot of different things. So I merely went along. I started to pursue watercolor, did that for 20 years. And I got, you know, I made some progress. But then, you know, something just in my work felt like there’s something missing. So I decided that, after our boys grew up, we had two boys and they were going off on their merry ways, so I thought, “Well, maybe it’s time for me to go back to school.” So what I did was, I enrolled at the University of Montana School of Art. At first the idea was, I’m just gonna take some art courses and try and fill in these gaps that were in my self-taught background as an artist. I then ended up taking 30 studio credits because I didn’t have an undergrad in art. Then I thought, “Well, I think I’ll apply for the grad program, for the MFA program.” And they accepted me and I got in.

That was really fun for me. That was kind of– I wanted to talk about the inspiration for Ubiquitous. After I got my MFA, I taught through the university for a little while. I participated in several exhibits. But I think what really happened that really got this going for me was that I started to work with Elizabeth Fischer, Olivia Steele-Mortimer and Anita Mora, and they provided me with 100s of their electron micrographs, something I’d been wanting, as Byron could tell you, they’d been something that for the longest time, I kept saying to him, “I really wanna see these electron micrographs” because I knew how amazing they would be. And they, these women shared them with me over 100s of different both transmission electron micrographs as well as scanning electron micrographs. And this lab is a level IV facility. This is a picture of the Rocky Mountain Lab in Hamilton, Montana, where we live, where Bryon works. Given that they study over 30 of the world’s deadliest pathogens, being an artist and being able to look at these was pretty amazing. For example, these are images of scanning electron micrographs that I was given, and salmonella, and then in the middle are the prion fibrils that Byron was just talking about.

Ebola on the right. Scanning electron micrographs are kind of like looking down. It’s like a topographical view. And then we have a transmission electron micrographs which are kind of like MRI, where you’re doing a cross-section. And that’s why these look so different. Pneumonia on the left, tick borne relapsing fever, poliovirus. After looking at these images, at the same time, my neighbor was a silversmith. I had always wanted to learn how to work with metals. So he taught me how to etch copper and brass. And then with this imagery in my head that I’d been looking at all these electron micrographs, I thought, “Well, why don’t I just etch some copper and brass?” So I did that and I submitted this particular piece to the Tacoma Art Museum, and it got into the Northwest Biennial. That was 2012. Little bit later, did one in brass. Some of the pathogens pictured here, amyloid, chlamydia, HeLa cells with chlamydia, Ebola, amyloid Ebola. As you can tell, I was kinda like a real fan of Ebola. I mean, it sounds really, really horrible to say that. But boy, it’s good looking. [chuckles] I was just really attracted to the aesthetics of Ebola. Anyway, so what happened in 2012 was, RML scientists and local artists got together and decided to do an exhibit together, probably the first time ever at the Ravalli County Museum in Hamilton. I had this piece there, I had my drawings there. And Byron is also an artist, which you may not know.

But these were some of his sculptures that he showed there. He worked in glass, he worked in wood. And, you know, he works with a lot of, like, found materials as well. So some of the wood that was in our property. These were just an example. I also was inspired by what was happening in the news. There was rarely a day that went by, you know, 2012 or so, that you wouldn’t see something in the newspaper about the next outbreak, pathogens, diseases. And I just found this to be, like, kinda remarkable, like, why is this happening? This one, Delays in Global Disease Outbreak Responses, H1N1, Zika. And then this particular article about an outbreak in Nigeria due to children not being vaccinated for polio, you name it, it’s kind of in the news. And then this particular image, which is kind of really sad to me because I love cows. I grew up in Wisconsin. And here are these dead cows. They’re being incinerated because they had mad cow disease. The reason this is important to me is because here’s an image of when we lived in London. We lived between King’s Cross and Russell Square. That little arrow is circling one of our favorite places to eat. And that happened to be a place that’s, it was a Greek restaurant, it served doner kebabs. Well, that’s ground up brain and spinal cord tissue. So, yum. [audience chuckles] So we just ate that because, you know, it’s cheap and we were poor and I didn’t have to cook, [chuckles] most importantly.

So then, by 1984, we left and we moved to Montana. And then the first evidence of mad cow disease broke out. Then other incidences where we had pathogens in our family would be, Byron was riding his bicycle in France and he was drinking out of the fresh streams of water. He contracted Giardia. And then he went to Nigeria and did the same thing and he lost about 20 pounds. Our son Evan had done a two-year international baccalaureate in India and he contracted Giardia as well. And he’s never been the same since. I can tell you that much. He’s really tried to figure out what it is, but it’s pretty tough. Basically Red Cross also does not want our blood any more because we lived in London during this time. We go to the Red Cross and wanna give them our blood and they’re, like, “No, you know, thanks but no thanks.” The other thing that really inspired me would be, we’d be on the plane. You pull out those magazines that are in the pouch in front of you. I would flip to the back where all the maps were because, I’m not a map person but I do love those flight patterns. I found them to be really fascinating. It just spoke to the fact that, you know, we’re so global now, there’s no place we can’t go. We can go anywhere. And I noticed several images. Here’s another one. Well, these images really stuck in my head.

And here’s another one that I saw online. I mean, to me that’s really beautiful. It’s just something, an image that once you see something like that, it’s inspiring. So, Ubiquitous: Migration of Pathogens began with the electron micrographs, like I mentioned, the flight patterns, and then what was going on in the news. And I think as an artist, you get sort of these things that happen where you’ve got information coming in from different places and you start to put it together and you wanna sort of express yourself artistically. So I had this idea to make a proposition to the Missoula Art Museum. I submitted 20 slides, realizing that usually when you submit to a museum, you’re not gonna get in, that’s kind of a given. It’s very competitive. You know, they get, like, 600 entries a year and you’re, “Well, what the heck. I’ll try it anyways.” I had about 19 slides of work that existed. But then, just to sweeten the pot a little bit, I threw in a slide that was fictional, one that I fabricated. And it’s, like, “You know, why not make it super complex? Let’s not just give ’em something that’s not very interesting.Let’s throw in some nice etched glass and some type of map behind it that suggested that pathogens might migrate through society.”

And it was a large-scale piece and I submitted it and kinda forgot about it. And then I got a letter in the mail, saying, “You know, we really liked your exhibition proposal, especially that one slide that you submitted [audience chuckles] of the large glass panels.” It’s, like, “Oh!” The initial reaction is joy, for about, I don’t know. How long did that last? Maybe 20 minutes, and then it starts to dawn on you that you have to actually produce this now. And I did not know how to etch glass, I didn’t know, I did never work on anything that large scale. And so I had a dilemma on my hands. This is what happens. I often, like, put myself out there, [chuckles] thinking that, “It’s not gonna happen,” and it happens. So, have to kinda rise to the occasion. So what do you do? Well, I needed to really understand what I was gonna talk about. I had these ideas, the visual ideas in my head. But, you know, how was I going to actually express these ideas? So the concept of migration of pathogens became the biggest challenge since grad school. I needed information. I already had the inspiration, but I did read some books that were quite helpful to me, “The Coming Plague,” which is all about newly-emerging diseases, by Laurie Garrett, and then “Deadly Companions: How Microbes Shaped our History,” by Dorothy Crawford, just a lot of great information. I was taking notes, trying to ingest all this information. Well, to make a long story short, what this did for me was, in my mind, I was able to solidify what it was that I was, why is it that pathogens have now become so rampant and why are these diseases popping up everywhere? Well, you go way back in history, we used to be hunter gatherers.

At the time, we were in very small, isolated groups and the pathogens had a very hard time moving from one band of people to the other. You know, if they’d knock out a whole tribe, they had a very hard time moving on to the next tribe because geographically they were so far apart. And then we moved on to slash and burn agriculture, where trees were starting to be cut down, they wanted more space, they wanted to start to domesticate their animals, biodiversity decreased and people started to move around less. They started to sleep in their huts or their homes with their domesticated animals. And microbes basically had a field day. From that point on, you know, fast forward to today, now these pathogens, due to our great modes of transportation, they’re hitching a ride on new hosts, they’re defying the latest antibiotic, there’s this anti-vaccination movement, increase in population, climate change, if it exists, and then that causes them to have to change, adapt, mutate and evolve. It’s all about survival. So I started to think about pathogens actually from that point of view, like, okay, they’re in this to survive, just like we are. So this whole idea of, first of all, this exhibition did not have a title for the longest time. But I think after I read these books, the word ubiquitous kind of stuck to my mind.

So it became Ubiquitous. It had three main components. The large glass piece became called Ubiquitous itself. My idea would be to have three sheets of tempered glass, each four feet by seven feet. And then behind that would be something, I didn’t know what, some type of world map inspired by all those things I saw on the planes. The second part of it would be migration of man. So there’s migration of pathogens, there’s migration of man. Now, how I was gonna do that, I didn’t know either. [chuckles] And then the third thing I did know was that I wanted to do some drawings. That to me was the only part I really understood so far. I was gonna do drawings inspired by electron micrographs I had seen. I would draw them on ground glass and on sheets of mylar, creating several multiple layers. So that part was not difficult. Getting back to the etched glass, though, that was my first thing I had to conquer in my mind. So, etched the glass. The only thing I had etched at that point was a beer glass. And it didn’t go very well. [audience chuckles] Our sons have a company, I was trying to etch the logo on it and it, you know, part of the thing slipped. I thought, “If this were four foot by six foot sheet of glass, I’d be in big trouble.” So at this point I started to make some phone calls and I started to get a little desperate because I was explaining my project, that I wanted to etch pathogens on glass.

And I would get these kind of silent conversations. Nobody was really jumping up for joy that they wanted to take on this project. So after probably a couple months of making these phone calls, I did find one person in New Jersey who could do this for me, at least he said he could. So, okay, his name was Wayne. “Wayne, I’m gonna send you an image of what I’d like you to etch,” ’cause at that point I didn’t even have anything yet. And I, again, relied on these electron micrographs to inform the design. And I wanted to choose pathogens that were interesting. I mean, it had to look good. They had to be sort of different from one another, looking for ones that, you know, how would I compose the glass? So I’m pulling information from here and there and repositioning it. I wanted the glass to show pathogens that were really well-known to people, that they had unique morphology, aesthetically pleasing. So I did this, I sent it over to Wayne. And about a year later, they were delivered. [chuckles] This is, we lived on Roaring Lane Road, and that’s our house in the background. This gigantic freight truck comes, and there’s Byron, always ready and waiting to help me. [chuckles] And then it gets unloaded and it’s in our driveway now. You can see it’s three sheets of glass that are packed very carefully. And, you know, they’re unwrapping it. But we had a year before the exhibit was to open. So they just kinda sat in our front windows for a while. It just turned out that they slipped right into the front molding, and Byron kinda put some wood there to hold them in so they wouldn’t fall out.

And we looked at pathogens all winter. [Pam and audience chuckle] But what we got behind the glass? That still really, really perplexed me. But, you know, if the answer doesn’t come to you right away, you just kind of say, “Okay, move on to the next thing.” So. . . I did. I did not come up with a solution, though, for this until six months before the exhibition. I went on to “The Migration of Man” component of this show. I asked 30 people, young and old, if I could cast their feet in plaster. You’re probably wondering why I asked them to do that. Well, I had been an instructor teaching the fundamentals of sculpture, and we happened to be working on hand cast in alginate. So you mix up this alginate, which is kind of a squishy liquid. You put your hand in there, wait about two minutes until the mold gets a little bit harder, and then you kinda jiggle your hand out of there. And then you pour in the plaster and out comes a pretty cool replica of your hand. Well, I thought, “What about feet?” So I did ask about 30 people, young and old, of people I knew, I could cast their feet. I got a lot of strange looks. Most people said, “Yes,” but then some said, “I’ve got this foot fungus and I don’t really think I want you to cast my foot,” or, “I don’t like my feet, my feet are ugly.” “Oh, no, no, you can’t do my feet, no.” But then some said yes.

So, why? Migration, to me, our feet are kinda like the ultimate mode of transportation for us. Yes, we’re assisted by wheels and rudders and propellers, but without feet, it’s kind of hard for us to get around. That was where that kinda came from. And via our feet, we move through society, pathogens can hitch a free ride, they can survive in new environments, which lead to more virulent forms, and I chose white plaster. I didn’t paint it, in other words. I didn’t paint it different colors because regardless of race, religion, age, gender, size, we’re all susceptible to at least some pathogens. In this picture of some willing volunteers, a very good friend of ours and her grandchildren, granddaughters, are very happy to do this project with me. I went from all these, visited a lot of people, but basically you put your foot into this alginate liquid and you have somebody stabilize your foot while it’s in there. That was really hard when I had an infant. The mom made her infant, made sure that her infant was sleeping and that I made the water extra warm to make it speed up the process, and pulled out her little foot, and the mold came out so well that I was able to duplicate it.

That was really cool. But anyways, you put the foot in there and then you pour the liquid stone into the mold and then you peel away the alginate. And what you get is hyper realism. This is what we got. So I had these feet. And obviously they just stuck their toes in there. But now I had all these toes. I didn’t really know what to do with it. I didn’t know how I was going to present them. So I was in my studio one day and I had a frame leaning up against the wall and it had glass in it. It had a black frame. And I just laid it on the floor. I had about 10 feet. I put them on top of the glass, I turned out the lights and I started to see these shadows. I thought, “Okay, that’s really inspiring to me. I think I wanna do it that way.” So the reason I did it this way was because the shadows kind of mimic pathogens. They kind of look like these bizarre shapes. And I just found that to be very fascinating. You can see how realistic they are.

People had, like, a bunion or a corn, or if they had a Band-Aid on their foot, you know, it all came out to be just like that. So the third component, then, were all these drawings that I did. I probably did about 30 drawings and more in three different sizes. Again, some were just like a– What I did was, I took glass and I ground it with carborundum, which is a gritty power, with water, and you just do these circular motions and you make it kind of foggy. Then I started to draw on it. So it kinda felt like drawing on a lithograph plate. I drew on the bumpy side. So when you look at it from the front, you got the nice glass, it’s nice and smooth. But on the back side is the drawing. So here’s some others, chlamydia, Yersinia pestis was one where I took some artistic license. I morphed the bacillus into these black rats that are the carrier. That was one that I kinda played with and kinda liked. And then pneumonia. . . HIV, salmonella, herpes. Some had more of a watery background. Just kinda depended on what I wanted to do. Here is parainfluenza, Francisella and Ebola. So again, back to this last component that I was just so perplexed by.

I literally tried– I had all these ideas. I mean, I was buying carbon rods and I was thinking, “Okay, I could drill hole into the wood and I could have a carbon rod arching over,” I’m thinking about these flight patterns. But then I’m thinking, “What if those carbon rods come out and they start to just jump all over the place?” So I started to really think about it and I kept staring at this world map and thinking, “That’s been done before. I can’t use the world map. That’s just, you know, it’s too boring.” So I started to look at it from the point of view of the pathogen and I started to slice this drawing into many pieces. And then I just recombined it. What happened was, now you could actually see Russia from Canada. I purposely left some continents intact, but now the UK is closer to Mexico and, well, I guess you can kinda see some of the different countries. I tried to keep it so that it was recognizable as our world, but wanted to show that, you know, really from the pathogens’ point of view, it doesn’t matter really how we recombine the Earth because the points of departure now have merged with the points of arrival.

So our mode of transportation has now made travel the key factor in the migration of pathogens. That is what is out there now behind the glass. So here I am in Byron’s studio, using Byron’s tools and making sure that I put them back. [audience laughs] Doesn’t always happen. That’s our dining room table where I started to assemble things. Some pieces needed to have a little connection. Here’s how it first appeared at the Missoula Art Museum in 2014. It was, I don’t know, few months after Ebola broke out. So the coincidence of that was, like, “Well, it’s not my fault.” [audience chuckles] There’s some people who approached this exhibit kind of with a little bit apprehension.

I don’t know, I think that’s just what happens. But Byron made the bases. Here’s an up-close view showing you, again, the effect of the light passing through this etched glass was quite important. I love how they’ve done it here, with the lovely lights that come way out in front of the glass because in the evening, you turn them on and you get the same effect of these shadows that, what do they do? They kinda multiply and they multiply the impact of this feeling of ubiquitous pathogens. You might have only had ’em one time, now you’ve got ’em, like, multiple times, multiple angles. Here’s another– it traveled on to the Nicolaysen Museum in Casper, Wyoming. Every time it was a new location, you have different ways to exhibit it. But there’s a migration piece in the middle. They always cordoned it off so people wouldn’t trip over it. That was a problem.

They’re really worried ’cause it is tempered glass. And if you fell on top of the feet, [chuckles] would not be a good situation. There are the drawings on the left. And then I wanted to show you just up close what those drawings look like and how they’re framed, just because the drawings were actually set back, recessed, to give this feeling to the viewer like, “You can go ahead and look at Ebola.It’s not gonna hurt you. It’s behind a vault and it’s behind glass.” And it just, I don’t know, it’s one of those things where, it was also like a repetition of that rectangle. The rectangle of each of the sheets of glass, the rectangular format of migration. So there’s this consistency of design running through the exhibit. And here it is being shown at Johnson & Johnson. That was the place that it was before it came here to University of Wisconsin. They actually held on to the exhibit for a year, ’cause I didn’t have any place to really put it and they were willing to hold on to it for me. But they had the drawings and they had Ubiquitous, but they didn’t have the feet because they didn’t want people to trip over them. Here’s my website if anybody wants to read about that or anything else that I do. And so here are some installation photos. I really can’t express enough gratitude for all that went into this. I was called early on by Daniel Einstein and said, “You know.” I didn’t have anything in front of me. They had all the work in crates. And he’s, like, “Well, how tall are the glass panels? Because we have to reinforce the wall behind it and make sure that those pegs just match up with the wood behind it,” and I’m thinking, “Well, it’s like seven feet plus the height of the bases.”

So I think they nailed it. So anyways, that’s how it looks right now. Does anybody have any questions? [audience applauds]