Seven Questions....with Yours Truly

Last week, my SciBling Jason Goldman interviewed me for his blog. The questions were not so much about blogging, journalism, Open Access and PLoS (except a little bit at the end) but more about science - how I got into it, what are my grad school experiences, what I think about doing research on animals, and such stuff. Jason posted the interview here, on his blog, on Friday, and he also let me repost it here on my blog as well, under the fold:

Here at Thoughtful Animal headquarters, we're starting a new series of seven-question interviews with people who are doing or have done animal research of all kinds - biomedical, behavioral, cognitive, and so forth. Interested in how animal research is conducted, or why animal research is important? Think you might want to do some animal research of your own someday? This is the interview series for you.

I've asked friend, scibling, and trusted advisor Bora Zivkovic (twitter, blog) if he would be our inaugural interviewee. In addition to his extensive blogging here at Scienceblogs, covering topics ranging from biology and circadian rhythms to science communication, journalism, media, and blogging, he is Community Manager at PLoS ONE. He is also the organizer of the Science Online conferences, and series editor of Open Laboratory series, which features the best science writing from blogs.

In traditional Bora style, the interview is long, but I found it so interesting and some of the topics so important, that I've not edited it down.

BORA Triangle Chem Lab 06.JPG

Figure 1: Bora Zivkovic, in his basic research days.

To start out, could you tell my readers something about yourself? Could you tell us a little about your academic and/or scientific career trajectories?

I grew up in Belgrade, Yugoslavia (now Serbia) which, at the time was a gray, dirty, polluted city (it is MUCH nicer now) of two million people and where the only species around were easily identifiable: pigeons, house sparrows, crows, rats, stray dogs, stray cats, house flies, mosquitoes and cockroaches. Yet, I always loved animals. I went to the zoo whenever I could, to every circus that visited, and rode horses. I spent summers in the country, but nobody in my family had any naturalist expertise, and I had no books or field-guides, so I never learned how to identify various critters out in the field - it was a bird or a lizard or an insect, and that's where it had to stop, no details.

I soon took to biology, reading everything I could get my hands on - a rare book, an occasional magazine article. That was decades before the Web! Understanding that there was not much opportunity to do basic biology as a profession there at the time, I instead went to vet school. And continued riding and training horses, with an eye at becoming an equine vet.

The Balkan wars of the 1990s brought me to the USA. I spent the first two years working at a horse farm here in North Carolina where I had an opportunity to talk to a number of vets and to observe the difference between the two countries in this profession. At the same time, I finally had the opportunity to find and read hundreds of books I always wanted to read, from Stephen Jay Gould to Richard Dawkins and everything in-between.

My old love for biology was rekindled, and, realizing that in the USA one can do basic science while being a vet is not as glorious as back at home, I decided to change my focus. Being told at NCSU vet school that I would have to start from the beginning (I was almost finishing up back in Belgrade) just reinforced that decision. I did very well on my GREs and TOEFL, but I applied only to the three local schools (NCSU, UNC and Duke) as with a brand new child it would have been difficult to move out of the area.

A year later I found myself as a Masters student at NCSU in the Department of Zoology (now Biology). That year, UNC department of biology (being much smaller) did not take in any new graduate students. Duke accepted less than 10% of applicants and my transcripts baffled them so they decided not to take a risk on me.

I got into NCSU by taking two graduate-level courses I paid for out of my own pocket - Behavioral Endocrinology with Dr. John Vandenbergh and Biological Clocks with Dr. Herbert Underwood. I aced those two courses, they had the opportunity to see who I was, and that made my acceptance into grad school easy. I was completely and utterly fascinated by the Clocks course - a field I have not heard of until I saw the title in the course catalog when searching for classes to take - so at the end of the semester I asked Dr. Underwood if he would take me as his graduate student.

You can read more about my equestrian past here and a more detailed pre-history of my biological/scientific trajectory here.

You mentioned some important differences in veterinary medicine, as a profession, between here and at home (in Belgrade) that led you to opt for basic science. Could you expand on that a little bit?

I always personally preferred basic science to applied. But in Yugoslavia back in the 1980s, there was not much one could do about basic science - there was a little bit done at the University, a little bit in Institutes, but there was preciously little funding, infrastructure or support for this. And not much freedom to choose the projects. Without test-tubes and reagents, people were sitting around the laboratories drinking coffee and gossiping (it is getting much better today, I hear). On the other hand, being a vet was fun for someone who loved animals. Especially if I was going to become one of the few sport equine specialists in the country, I would be making a nice living, have huge respect from the people, and generally have an interesting, intellectually and physically challenging, and good life.

In the USA, veterinary medicine is hugely competitive. With a diploma, one does not have much choice as far as where in the country to live and work. It takes some serious money to start a practice. And there is a litigation culture that forces one to practice defensive medicine instead of the art of treating animals. On the other hand, even in the middle of economic crisis and overproduction of PhDs, the USA is still by far the best place on Earth to do basic science - there is a degree of intellectual freedom, a level of infrastructure available, and in general a meritocracy-based system well above that of any other country, especially in comparison to a small, ex-Socialist one.

Some people decide that they want to work with animals, and then find a particular research program they like. Others are interested in a particular empirical question, and discover that animal research is one of the best ways to approach that question. Why did you decide to become involved in animal research?

It wasn't a conscious decision at all. It is just the way the world worked back then! Let me explain...

I came from a vet school background. My strong background in physiology, as well as experience with working with animals, were strong points for my acceptance into the program, especially as nobody could figure out what the transcripts from a completely different educational system meant.

When I joined the Department, back in 1994, molecular biology was something that was done elsewhere - in departments of genetics or biochemistry or microbiology. Everybody in our department worked with animals. That was an "of course" kind of thing - only one or two faculty at the time routinely ran gels. The internal division of the department was between people who worked with animals outside (ecologists) and those who worked with the animals in the lab (physiologists). A few of people did both: an ecologist who occasionally brought animals into the lab to do behavioral experiments needed to understand what they were doing out in the wild, and a physiologist who kept some of his animals in natural or semi-natural enclosures to test some of his hypotheses for which a purely lab test could be misleading. People studying fish often did both lab and field work as well.

Of course, many labs, while doing purely animal work, also did quite a lot of tissue culture, cell culture and hormone assay research as well. Animals were both the subject and the object of study, not just sources of DNA samples. People in the department directly worked with their own animals and took care of them themselves. As many different species were studied, those PIs (and their students and technicians) were also the world experts on husbandry of their animals and would not risk having their animals stressed out by some faceless techs in some faraway animal facility. Only some of the mice and rats, rarely used in our department anyway, could be entrusted to outsiders as there were well defined methods for their husbandry. Every other species (especially things like venomous snakes!) was best kept by the lab itself in order to make sure they get the best and most appropriate care.

My interest was in evolution, but my background and strength were in physiology, so doing physiology in grad school (while perhaps thinking about the data in evolutionary context) was a logical plan for me. Circadian field is exciting, and Japanese quail (the species of bird used in the Underwood lab) was a rarely used and intriguing animal model for study of some particular aspects that are (or were at the time) difficult or impossible to do in mammals. So, on Day One, I started learning how to take care of the quail and how to work with them. It never occurred to me at the time that there was a way to study biology without working with animals - what's the point?

You also need to go back in history. In 1994, the field knew of period mutants in a couple of protists (Euglena, Pseudomonas) with no molecular characterization. We knew about the tau-mutant in hamsters, but had no idea what was the gene in question until much later. The only two genes involved in circadian rhythms that were known at the time were period in Drosophila melanogaster and frequency in Neurospora crassa. It was the pre-history of molecular chronobiology, not much was possible to do with techniques available at the time, and most circadian labs (including ours) were not equipped to study genes in any way: no know-how, no experience, no equipment, and frankly, no interest.

It was in 1995, a year into my research, that the second fruitfly clock gene, Timeless was discovered, as was the first mammalian clock gene Clock. These were subsequently followed by discoveries of many additional genes in a number of species, including cryptochrome, bmal and doubletime in animals, wc in fungi, toc in plants and kai in cyanobacteria. Many of those early studies were crude and, so excited about the gene discoveries, ignored the available understanding of the way clocks work, oversimplifying the role of these genes to the point of being just pure wrong. The excitement of these discoveries brought people from other fields into chronobiology and their first few papers tended to contain beginners' mistakes in experimental design and data interpretation due to their ignorance of the field. Too much excitement was there about the new shiny objects, the genes discovered left and right at high pace, and not sufficient understanding how they fit into the system as a whole. It became a bandwagon, and I was not interested in jumping on one. I was much more interested in being well known and respected as "the quail guy" who does solid work at several different levels of organization (of which molecular level could be one), incorporates ecological and evolutionary thinking into the physiological work, and does not make rookie mistakes, than in being one of the thousands studying boring and almost irrelevant minutiae of how period interacts with timeless. My bias, my personality, but also the Zeitgeist of the time at which I entered the field. The world has changed since then....

What have been some of the most interesting or challenging projects you've worked on, in the course of your animal research?

USA was new to me. Graduate school was new to me. Basic science was new to me. I had no idea what to expect when I joined the Underwood lab in 1994. And some things I learned only years later....

For example, I had no idea that there was a longish list of people who lasted less than a year in the Underwood lab. Dr. Underwood was selecting for self-starters. At the beginning, he gives you a copy of the grant (so you get the idea what is your and other's work all about), a small stack of previous papers from the lab (to learn the background), and an experiment to do. Apparently, many people did not understand they were supposed to actually do the work. Dr. Underwood would probably gently ask them a few times per year if there were any data to show yet. If there were none by the end of the year, that was the end of their presence in the lab.

I was excited about diving straight into research. I read literature in the field going all the way back into the 19th century - probably every paper up to about 1995 when the molecular field exploded and produced far too many (initially mostly shoddy, until people learned) papers to keep up with. I also never paid much attention to the medical side of the field or pretty much any research on humans. But I read non-circadian papers on Japanese quail, and on bird behavior, and hormones, and evolutionary theory, and other tangential topics.

BORA Quail and I 4.jpg

Figure 2: Bora with two of his Japanese quail.

I took care of the quail, visited them every day, downloaded the data once a week and showed them to the boss. After a couple of months, I started putting in my two cents as to what the data meant. Another couple of months and I started suggesting we slightly change the experimental protocol in order to better capture what we were seeing in the data so far ("OK, do it that way from now on"). I started suggesting additional experiments ("OK, do it") and we started having interesting discussions at the white-board. Herb Underwood is a serious guy (and has a big beard that hides facial expressions), so I could not get any vibes from him if he was happy with my work or not for at least the first year or so. Thus, I worked harder, just in case... and I was taking the opportunity, being in the USA, to take double the load of classes than expected, including courses in dinosaur osteology, avian biotechnology techniques, and history of science. All that while my first kid was very little, second kid was just born, I was TA-ing various anatomy and physiology labs, and my wife was also in school - we traded the kids between my and her classes and I did much of my research at night and on weekends.

Another thing I did not know at the time was the ranking - where did my PI, my department and my university stand within a national or global picture. I learned some of that stuff a little later. In 1995, then departmental chair did the citation analysis of all the faculty - the result was Dr. Underwood up in the stratosphere, a couple of others kinda middlin' and everyone else essentially nowhere (I have to note here that the Department has incredibly improved in the intervening fifteen years, hiring brilliant young researchers for every open position - the difference between it today and 15 years ago is like night and day. The reasons are hard to figure out, but then new university President, Maryanne Fox, though far from being loved by everyone, changed the atmosphere at NCSU - suddenly everyone lost the inferiority complex of an ag/tech state school and started thinking of the school as a top research institution, thus boldly offering jobs to the top researchers instead of "whoever we can get". Also, foundation of the W.M. Keck Center for Behavioral Biology changed the tone of the Department in a positive way, encouraging top-level collaborative work by everyone.).

When I went to my first conference a couple of years later, to the meeting of the Society for Research on Biological Rhythms (SRBR) I was in awe of all the Big Names whose papers I have read and mainly spent time with other grad students. Of course, the first question is "Who's your PI?" "Herb Underwood" "Wow! Lucky you! Great advisor!". Apparently, and I had no idea until then, my boss had quite a serious reputation in the field.

When you do your work and publish your papers and do not really know how it stacks up against what other people do - understandable confusion at the beginning of one's graduate career - it is nice to get some feedback from others every now and again. Only much later did I realize that it was highly unusual to publish five papers without having to go through multiple rounds of review. In those five papers, we only had to change one word ("biweekly" to "twice a week") in one of the manuscripts. The first review I ever saw on our first manuscript said something like this "As expected from the Underwood lab, the work is excellent and writing clear. Discussion section is long, but should remain as is, as it is a good review of the topic for the newcomers to the field". And that was it, the entire review! I had no idea how rare this was until years later, when I started reading horror stories on science blogs!

In 1998 I defended my Masters Thesis (also the year when I became a US citizen and cast my first vote). My defense was 50 minutes long and some people in the audience thought it was a PhD defense - there was so much data. I got two long first-author papers out of my Thesis, plus a paper on the work we did in the lab together (not part of anyone's thesis) and a review. Many years later, when I started blogging, I revisited my old papers and tried to translate them from scientese into regular English. I have summarized the review here, my first first-author paper here and our collective paper here. My second first-author paper resulted in two blog posts - one straightforward summary here and a look at the same data from an ecological perspective here. That latter post resulted in more research being done years later - I recently blogged the results here.

At about that time, Chris Steele joined our lab as a second PhD student. Up till then, I was alone - it was just Dr. Underwood and myself. No technician, no undergrads, nothing - the last PhD students, a postdoc and a technician were done and left shortly after I arrived at the lab. At about the same time, we have moved our quail from a lodge outside of town to a brand new animal facility on campus. I had to dismantle our setup, removing hundreds of wires, and then reassemble and rewire it all in a new lab. Plus I built ten new boxes/cages with all the wiring and other associated stuff. I became quite a carpenter and electrician over a few months. With so much electricity in the new building, it took us about six months of troubleshooting until we managed to eliminate all the interference we were getting in our radiotelemetry system.

With Chris in the lab and the increased capacity, our work really took off. Chris and I became fast friends and spent a lot of time together on and off campus. We hired some excellent undergraduates to help with research (and some are now making their mark in science as graduate students in big research institutions - having a letter of recommendation from Dr. Underwood opened a lot of doors).

For about a year, after just getting a new grant funded, Chris and I did everything together, starting a gazillion new experiments, but not yet differentiating which experiment will be in whose thesis. At the end of that year, Dr. Underwood asked us to divvy it up. We looked at each other. Chris said "I'll work on males", I said "I'll work on females" and that was it. This actually makes much more sense than you think - males were used for the study of the clocks and melatonin in the brain, pineal and eyes, without any cycling sex steroid hormones introducing noise into the data. I was using females to see what were those steroid hormones doing to the clock, if there was a clock in the ovary itself, and how all that affects photoperiodism (use of changes in daylength to trigger seasonal events). So this division was logical. Chris' work resulted in three more papers, one of which I was a co-author on. I blogged about two of his papers here and here.

My Dissertation was supposed to contain a series of experiments looking at the effects of sex steroids on circadian function. I did that, but I also ended up doing four times as much - a follow-up series of experiments to my MS work on photoperiodism, a collection of experiments that crossed some t's and dotted some i's on previous research, and a series of experiments that came to me out of the blue and make quite an interesting story:

Obsessing over my own data for a few years, knowing the quail clock literature backwards and forwards, being very familiar with the literature in two other unconnected fields, and most importantly personally taking care of and observing my animals every day for many years, I made some observations and started putting together 2 + 2. I had a hunch, just a hunch, but it sounded plausible to me. No money to do it, and no preliminary data to write a grant proposal for it. Not to mention it would be a huge distraction from the work I was supposed to be doing. So I decided to come up with preliminary data by piggybacking on the work I was doing anyway, thus not doing anything extra that IACUC (Institutional Animal Care and Use Committee) would be unhappy about. I was breeding and raising quail anyway - I could count them and weigh them without getting a special approval for it. I could do my regular experiments but extract additional information out of the data. Instead of just watching them, I could write down every instance of a behavior and do stats on that. And it appears that my hunch was right. But I can't yet tell you anything about it because none of that is published and I am still hoping to publish one day.

After ten years of grad school I was getting exhausted and burned out. My father, my grandmother and my grandmother-in-law (with whom I was close) died one after another. I got depressed. I stopped doing experiments and was writing my Dissertation. Every week I showed my progress to the boss, each week having less and less to show. One week, as I made no progress on writing at all, I decided to finally show him my secret, backroom, hunch-based data - a good 45-minute PowerPoint chock full of data. His eyes got big, his jaw dropped, he made sure the door was closed and he ended saying that THIS was my Dissertation and everything else belonged to an Appendix. But even that could not save me from the depression. Living in the Bush nation after 9/11 was depressing. Starting to read political blogs was depressing. Being all macho and not admitting I had a problem was not helpful either.

And not being excited about any of the potential post-doc positions was not helping either. Everyone was telling me that I needed to do a post-doc in a molecular lab, to learn those techniques, in order to be able to "sell myself" later. But I did not care about it much. I was interested in animal behavior. Perhaps getting some of the new neuroscience techniques was going to be a better strategy. Eric Herzog was interviewing in our department and it would have been great if he moved here. It would be weird to do a post-doc in the same department, but just his presence would cheer me up (if you don't know Eric, nobody could NOT be cheered up by his presence), we could perhaps do an experiment together, perhaps he could help me motivate myself to get the dissertation done. But he took a job elsewhere and it takes some time to move and set up the lab and get started, and that would have been too late for me. Mike Nitabach (at Yale) did not move into chronobiology yet - his work is exciting and cutting-edge and if he was already in the field at the time, I would have tried to go there for a post-doc. Rae Silver came to visit one day and told us that, due to the funding situation, she has stopped her exciting bird work and focused only on mammals, i.e., the stuff I did not care about. I visited Russell Foster's lab at Imperial College in London, but he was ending the exciting work on mole-rats and moving the lab into some hypermolecular direction I did not find that interesting.

A cancer lab nearby was starting to delve into circadian rhythms and we wrote a grant proposal together, including some pilot studies on clock gene expression in quail. The grant was not funded, but I went to that lab anyway (we did not even have an electrophoresis apparatus) and did a series of experiments there to see how it feels to run gels. I learned I was really bad with the pipetter! I also learned that, despite their assurances, I did not trust my own data: too many steps, too much blind recipe-following from the kits, too much trust that all reagents are doing what they are supposed to - and the data were showing exactly what I was expecting (and not what a couple of other quail papers recently published were showing) which made me suspicious - it was too good to be true. The whole process made me unhappy.

Finally, the hyper-competitive atmosphere in the lab was a real turn-off. Everybody working in silence, 13 hours a day, six or seven days a week, getting pale like the Eloi from the Time Machine by H.G.Wells. One week I attended a lab meeting of the circadian sub-group of the enormous lab, on Friday afternoon, ending at 5pm. One of the post-docs showed some data, explained a band on a gel was an artifact, and suggested a way to do the experiment without getting that artifact. The PI told him to show him the clean data first thing Monday morning.


Nobody tells me when, where and how to do my work. I may choose to work nights, weekends and holidays because I do best work when there is nobody else around (I am hypersocial and hyperactive, so when people are around I like to talk instead of work), but that is only my choice. I have never experienced that attitude in the Underwood lab (or for that matter any lab in NCSU Zoology). Circadian research is very slow - one day, one data point - and it has its ups and downs. As long as I had some data coming in at some times (and doing several experiments in parallel helps - at least one may be working at any given time), avalanches of data punctuating dry periods of failed experiments, I was fine. Dr. Underwood never gave a deadline, certainly not such a short one, and certainly not one that forces me to work over the weekend against my will. Balancing work and family was always understood to be important for grad student's mental health (and thus productivity) in the Underwood lab, and made everyone happier. This experience in the cancer lab certainly did not do anything to brighten my mood and kick me out of my depression. I was looking forward to a post-doc as a stage of increased, not decreased autonomy in my research. Apparently, that is not how the world works, and I was just starting to learn that.

So I got out of science and became a science blogger instead. One wonders what could have been if I defended the Dissertation, survived a postdoc, and got a job in academia. Those are fun fantasies to have. But I do not regret the move.

The use of animals in research is (to put it lightly) fairly contentious. If you feel comfortable discussing it, we'd love to know your thoughts on the issue.

Not being involved in any research any more, it is pretty easy for me to discuss this topic, and I have done it in the past - see this blog post for an example of a discussion about Animal Rights and this old post about my views and experiences in animal research (both also link to some other posts of mine on the topic).

Molecular biology proposes, animal research disposes. Or: molecular biology results in hypotheses, animal research tests them. Biochemistry, molecular biology and cellular biology discover fascinating details about the workings of molecules found in a cell. But all of those mountains of data are just hypotheses, to be tested in whole organisms. Every molecular finding is a suggestion that a process MAY work a certain way in the organism. Only whole-organism research can tell if that process actually works that way in the real world.

Until tested in whole organisms, data from research in a dish or on gels are chemistry. Once confirmed in a whole organism, those data become biology. Why? Because biology is a science that studies organisms, their adaptations to the environment, and the evolution of those adaptations. Knowing the internal parts (including all the way down to the level of molecules) and how they work in a dish is helpful only inasmuch it helps to answer the organismal questions. The same logic applies to biological questions relevant to human medicine - it is a human who is sick - though I personally do not have much interest in medical questions. Whatever one's personal interests are in biology, organismal research is essential.

The way a bunch of molecules play with each other within a cell in a way that makes everything in that cell cycle with a period of around 24 hours is incredibly fascinating! But early molecular papers made big leaps from what a bunch of genes in a single cell did to the rhythmicity of the whole animal. The increasing understanding of complexity of the molecular mechanism, the better understanding of inputs that reset the cellular clock, the better understanding of outputs from the cellular clock that time all sorts of events, the realization that every single cell in the body has a clock, and the better understanding of the way clock cells "talk" to each other, have gradually much improved the literature on the molecular basis of circadian rhythms precisely because those findings force the thinking away from a single cell and the molecules in it, and up to higher-level processes, e.g., how clock cells work as ensembles in tissues, and how different tissues and organs affect each other's timing, and how the external environment affects the circadian system, and how did such a system evolve as an adaptation. And that brings us up to the level of the organism again, and the necessity of studying the system in the whole animal.

What is important to understand when studying timing in biology is that the identities (sequences) of molecular players are not so important. Cyanobacteria, plants, fungi, animals - they all use completely different molecules to drive cellular and organismal rhythmicity, implying independent origin and evolution, yet identical function. What is important is to study the rules by which they operate, often best understood with mathematical models,
e.g., Phase-Response Curves, Limit Cycles, etc. How does an environmental cue (e.g., light) reset the clock, how that affects the synchronization between clock cells in a tissue, how that affects synchronization between organs and organ systems to produce a coherent biochemical, physiological or behavioral function? Knowing the identity of the molecular players is very useful as a tool - if you know the sequence, you can attach a dye or something that glows in the dark to one of the clock gene promoters and thus measure and visualize rhythmicity in cell cultures, tissue cultures or whole organisms. Modifying the experimental environment then helps us to uncover the rules by which the elements of the complex circadian system operate.

In mammals, most of the circadian system is driven by a single pacemaker which is tiny - just a few thousand cells at the bottom of the brain (so-called Suprachiasmatic Nucleus). It is very difficult to manipulate or study various sub-populations of those cells in live, freely moving mammals, to see what rules they use to interact with each other in producing a coherent daily output. But in non-mammalian vertebrates, like birds, there are multiple pacemakers, and some of them are quite large and easily accessible, e.g., the pineal organ at the top of the brain, or the retinae of the eyes. Those organs are easy to manipulate - one can remove them one at a time and see how the rest of the system works when one or more pieces are missing, or one can cut nerves connecting them, or one can manipulate (pharmacologically) candidate signaling molecules (e.g., melatonin) between the clocks, or prevent the light from reaching one or more of the clocks by placing black opaque patches (if over the eyes, they look like pirate patches!) either continuously or putting them on and off every 12 hours, etc, and from such experiments figure out the rules. This is the kind of stuff we mainly did in our lab (plus some hormone assays, and later immune assays and tissue culture) - as you can guess, some of that stuff is quite invasive! Which brings me back to the question of animal research: it's complicated. I am certainly not advocating unregulated, painful animal experimentation just because a researcher has scientific curiosity which can only be satisfied that way. We have to decide what is and what isn't an acceptable way to treat animals in research, while knowing that agreeing on NOT doing some kinds of experiments may mean we voluntarily accept that we will NEVER know the answers to some scientific questions. It is a tough decision to make, every time.

If you have read The Immortal Life of Henrietta Lacks by Rebecca Skloot (and you should read it - it is an amazingly good book), you have learned about the evolution of ethics in research on humans over the past several decades. Society changes. Science changes. Our ethical norms change. Taking a piece of tissue from a patient back then meant nothing more: taking a piece of tissue, essentially garbage for the patient, potentially useful material for medical research that may save lives in the future. But today, that piece of tissue can be ground, the DNA extracted and the genome sequenced! Whoa - that piece of tissue is not just a piece of tissue any more: it now contains much more extractable information about the patient it came from, perhaps information that the patient does not want to share. So you gotta ask for permission first, and learn to take a No for an answer! And realize that some solvable questions in science will, thus, never be solved, and that is OK.

It is likewise in animal research. Societal sensibilities toward animal research change over time. Nobody can today do the kinds of experiments that used to be done back in the 19th century. And that is good. But if those experiments were not done then, we would not understand biology well enough to devise techniques that make such experiments not needed any more today. As in everything in history, it is a mistake to judge people in the past by our current cultural, moral or ethical standards. They did what was ethical at the time. The same stuff is not considered ethical any more, so we don't do it. But luckily, our predecessors could do it, and did it, and on the shoulders of those giants we built the edifice of biology which incorporates knowledge that makes those most ruthless examples of animal experimentation obsolete. They were necessary to get the field going, but they are not necessary any more, even if that means there will be some things we will never get to know.

In our lab, we used to spend about six months discussing every experiment that we may want to do before deciding if it was worth doing and if we were certain that we have perfected the experimental design in a way that will maximize the usefulness of the experiment and minimize the number of animals used, minimize the pain, and maximize the benefit we gain from doing the experiment. Those experiments already got NIH funding, which means they were already approved by the IACUC (Institutional Animal Care and Use Commitee) as NIH requires the IACUC approval to be sent attached to the grant proposal. If we decided to make modifications to the procedure, we would have to get the IACUC approval again, which often took a long time. But the rule was never to touch an animal until there is an IACUC green light. If they say No, that is the final word on it. We may cuss and curse, but we turn our attention to other experiments instead and try again.

In the USA, regulation of animal research at the federal (and state) level is older (and one can say more mature and better developed) than regulation of research with human subjects. Regulation of research on humans got enshrined into law due to some horrendously unethical studies that came to light, including the Tuskegee syphilis experiment. The existence and activity of IRBs (Internal Review Boards) is essential in preventing such inhumane research from happening again. But there is a flip-side to this as well. IRBs have become bloated bureaucracies that are delving in areas that are not strictly ethics. Apart from costing a lot of money, they often delay or eliminate useful studies, steer students away from research with human subjects and, most dangerously, force researchers to modify protocols in ways that make studies more dangerous and less ethical than initially proposed. This trend - IRBs making decision in areas, and at levels, well beyond federal and state rules, with deleterious effects, even has a term: "Ethics Creep". It is extremely instructive, eye-opening and gasp-inducing to read the few studies of this phenomenon published to date, e.g., Ethics Creep: Governing Social Science Research in the Name of Ethics, by Kevin D. Haggerty, Qualitative Sociology, Vol. 27, No. 4, Winter 2004, Ethics Creep, Soft Law and Positivism: The Problems of Regulatory Innovation in the Governance of Human Subjects Research, by Igor Gontcharov, Subjects Research (October 2, 2009), The Dysregulation of Human Subjects Research, by Fost and Levine, JAMA.2007; 298: 2196-2198., and Mission Creep in the IRB World, by Gunsalus et al. Science 9 June 2006: 1441 (followed by a couple of Letters to the Editor in the September 8, 2006 issue). It has even touched journalism, especially when done in Universities, by j-school students - see Journalism and Institutional Review Boards, by Leon Dash, Qualitative Inquiry, Vol. 13, No. 6, 871-874 (2007) and Student fights research board, by By MARY JO FELDSTEIN, Digital Missourian, May 13, 2001 for an example.

I am not aware of any studies on the occurrence of Ethics Creep in regard to animal research. Yet everyone in the field is well aware it is happening - and happening very fast! Kinds of experiments I did in my first year of grad school, in 1994, I could not get approved at the end, around 2003. My advisor, after I left the lab, submitted a small proposal for a substantially less invasive experiment and got a resounding No from the IACUC. This being a clear signal that no research can be done in the lab any more, he packed up the entire lab and sent all of the equipment to a young researcher in Pennsylvania, and kept teaching for the remaining couple of years until retirement.

When I just started in 1994, we did surgeries on a lab desk, the anesthetised bird secured to the desk with masking tape, we cleaned our tools with alcohol swabs, and had never had any infections or other problems - the birds' immune system is just too strong for such stuff to occur. But then, a few years later, IACUC forced us to switch from injectable anesthesia (ketamine/rompun mix) to inhalation anesthesia under completely sterile conditions. My wife, who is a nurse, laughed very, very hard when she saw pictures of me wearing a surgical mask, a hat, and sterile gloves. And I had the tiny (100g) bird all set up with an inhalation mask on a sterile field! Quail is poultry. That level of sterility and anesthesia is not used in human surgery! Actually, the "old" system, injectable anaesthesia, was faster and we had no mortality at all. But with inhalation surgery we suddenly started seeing birds die, as the bird remains anesthetised too long (while the ever-present IACUC vet is slowly checking every step and talking through it) and the dosage of anesthetic is tricky. Did they change their mind? Yeah, right! They are omniscient, and the actual well-being of animals is far less important than the high they get from exerting power over people who are better educated than they are.

BORA preparing for sterile poultry surgery.jpg

Figure 3: Bora, preparing for sterile poultry surgery.

But IACUC (at least the one at NCSU I know, though I heard many similar horror stories from other Universities) has nothing to do with reason, science, or investigators' experience. They often insist on practices that are actually worse for animals' well-being than what the investigators, who have spent decades studying the particular species and honing the best, least stressful husbandry methods, suggest. They are petty bureaucrats who don't know anything about biology and misguidedly apply human aesthetics to their decisions. Because modern Americans prefer everything spotlessly clean and sterile, the IACUC members insist the same for animals even when that is exactly the wrong thing to do. There was a researcher on fish who was told by the IACUC to make the water in the tanks crystal clear. He did not know whether to laugh or to cry - that species spent almost 500 million years in murky water, the water in the tanks was purposefully made murky, as in clear water the fish would probably die of stress - but No, the IACUC does not listen to reason, they insist it's their way or highway. If we really wanted to make our quail happy, we would have provided them with sand boxes to use as dust-baths - in the field, quail bathe in dust, which helps against parasites like mites, and appears to be enjoyable to the birds. Dust!? IACUC was not having any of that. A bird is a feathered rat, and should be kept in sterile conditions, they said. Later, they wanted us to predict exactly how many males and how many females we will have in the future, before any of the eggs were even laid, collected and incubated! How could we know what the fertility and hatchability was going to be, or sex ratio? They were not going to accept an expected range - they wanted the exact number of animals.

I was pretty vague above about the membership in the NCSU IACUC. After all, several of the faculty are nominally members of it, smart people, themselves animal researchers. The problems really started when a new person was appointed the chair of the IACUC. He quickly ratcheted up the "criteria" for approval of studies, pretty much making sure that few if any proposals ever pass. He directly closed one lab for not following one of his nonsensical rules. Many others who could, quit animal research and moved to in vitro or molecular research instead. Those who had to keep working on animals, switched to mice or rats and left the entire care to the Animal Facility staff. Some moved from vertebrates to invertebrates. Some moved to other schools. Some young ones rejected professorial position offers upon hearing about the state of IACUC. The last graduate course still using animals in a teaching lab had to quit doing so in the end. One of the main motivations for me to do a study in an invertebrate on the side - the crayfish - was to get some experience (and perhaps even a publication) in a kind of animal for which an IACUC approval is not needed (they only control vertebrate research). This one person, over a period of about ten years, effectively eliminated animal research from NCSU - the PETA's wettest dream. There was an action at one point to get rid of him, even appealing directly to the university President, but to no avail - there is no formal mechanism for firing the IACUC chair, so he continued in this role for quite some time (he is not the Chair any more, but he is still an NCSU employee, not sure if he's a member of IACUC right now). Obviously, the system is vulnerable to the sensibilities of a single person, which indicates that the system is not robust. I would love to test and document this. If I had time and energy, I'd use Freedom Of Information Act and subpoena the entire IACUC documentation from the very beginning till today and do an analysis. I would love to show, graphically, and publish that somewhere as I know the data extracted from the documents would show this, that his arrival at State resulted in almost instantaneous and enormous drop in number of species used, number of individual animals used, number of experiments performed, number of proposals approved, number of grants submitted and funded, number of labs and people (P.I.s, students) using animals, number of papers published, number of Theses and Dissertations defended that use animals in research, and total amount of money this all costed NCSU over the years. Perhaps one day I will do that. Or someone should (with proper controls - comparing the sama data from a few other schools, where there may also be a decrease in animal research as a result of a molecular bandwagon effect, but much smaller and slower, with no particular date being an inflection point when the downward trend started). It will also be interesting to see if the numbers started improving after the new Chair was placed in that spot around 2006 or so, which would again indicate too much sensitivity of the system to actions of a single person.

I have a feeling that, because nobody is looking, Ethics Creep is happenning much faster and much more viciously in animal research than in the realm of research on humans, which is under constant public scrutiny. Also, it is hard to argue that clinical trials should be abolished, or psychological research. But explaining the importance of animal research to the uninitiated, especially if they have already been fed a diet of PETA propaganda, is an uphill battle. There is nobody arguing for abolishing all human research, but there are people who seriously argue (and get opportunity in the media for their stunts) for completely abolishing animal research.

What advice would you give to an aspiring scientist who would like to become involved in animal research?

Don't do it. You are a couple of decades too late to the scene. It's over (unless a bunch of heavy-duty, top scientific stars don't raise some serious noise, litigation if necessary, to bring the system back to reality).

One of the main reasons I am happy that I have left laboratory science is that I don't have to deal, in frustration, with an IACUC that prevents me from doing anything meaningful with animals - really the only bad side to an otherwise amazingly positive graduate school experience. If you gave me an appointment at a University right now, space, funds, everything I need, what would I do with that? It is an interesting thought experiment. Thinking back at all the quail experiments I was thinking about doing for the rest of my career, there are a number of them, especially the most invasive ones, that I'd be willing to let go of - we already know enough and I can live with us never knowing the answers to those particular questions. There are a number of other experiments, mostly not invasive at all (breeding experiments, behavioral experiments, field ecology, combined with some math modeling), that I would still love to do, or see done by others. But given space, time and money, I would almost certainly switch to an invertebrate model animal and, although this means learning about the model from scratch, would never look back.

Another solution, though probably just delaying the inevitable by a decade or two, is to set up a collaboration with a researcher in a developing country in which animal research regulation has not already reached the berserk levels of Ethics Creep. Let the collaborator do all the animal work, while you do the in vitro, in silico and molecular work and provide the funding for the whole endeavor.

Beyond the notion that having a solid background in science and research design allows you to better understand and communicate new research findings than your average science communicator, has your training as a research scientist been helpful as a science blogger or as Community Manager for PLoS ONE?

When PLoS opened up the position of the Online Community Manager, they were looking for a person who was both a scientist and also had experience online. People they interviewed tended to be strong on one, but weak on the other aspect, i.e., either scientists with little online experience, or bloggers (even programmers) without science education. At the time I got hired, I was really the only person who qualified on both counts - a scientist and a blogger. And not just a blogger, but a fairly well known blogger, well liked blogger, with a sizable reader community already in place. Plus I was already a proponent of Open Access and was regularly blogging about PLoS articles because I could read them online for free. That experience in managing an online community, behaving intelligently online, knowing how to engender trust, or diffuse online squabbles, is a much more important of the two skills I was hired for. My scientific background helps occasionally, when looking at media coverage, determining if a comment on a paper makes sense, or choosing upcoming articles to highlight. My connections in some areas of research (chronobiology, marine ecology, dinosaur palaeontology) brought us some submissions to PLoS journals, especially PLoS ONE, as well as some Academic Editors, which in turn opened up PLoS ONE to many more submissions from those fields.

As for science blogging and communicating, having at least some scientific background is practically a pre-requisite for being a science blogger. I am not that different from other scientists who blog - I tend to focus on areas of science I understand the best, as I can write about those from a position of expertise, as well as with confident clarity. As English is second language to me, it is understandable that my English is somewhat simpler than that of native speakers, which is a good starting point for good communication for lay audience. I always strived to communicate well, even while I was in grad school and did not plan for a career in communication. The papers from our lab, though written in scientese, are pretty clearly written, as much as it is possible when discussing arcane circadiana.

I think the question about this should be reversed - science blogging made me a better scientist. Reading masterful analyses of scientific papers by other science bloggers turned me into a better reader of the literature, and a more critical thinker. I so wish there were science blogs when I started grad school. I would have learned so much! It would have been easier for me, even very early on, to gauge how my research, my advisor, my department, and my school stack up in comparison with others elsewhere. I would have learned the differences between my lab and department and others. I would have appreciated even more the laid-back, relaxed atmosphere in our lab, in which the Boss comes to the lab and does stuff, and keeps his own intellectual curiosity alive all the way through retirement, and treats PhD students as equals. I would have heard much earlier on about the rat-race in more medical fields, and the post-doctoral horror stories from such labs. I would have learned, from blogs like DrugMonkey, how to prepare myself for a career in less laid-back schools, in cutthroat competition, how to write winning grant proposals and navigate the political and administrative aspects of academia that were easy at NCSU. And I could have started blogging myself, perhaps even try Open Notebook Science of sorts, piping automated radiotelemetry data straight to the Web! Who's gonna scoop me? Japanese Quail? Slow Circadian research? No competition to worry about, so why not do science in public?

Thanks for your time, Bora! I hope my readers find your thoughts and experiences and fascinating as I have.


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