Eduardo CesarFor the pioneer in renal physiology research in Brazil, Gerhard Malnic, 76, one of the pleasures of science was always the possibility of using his own hands, whether it was to manipulate and dose the fluid extracted from the minute tubules of the kidneys of animals in the laboratory and of human beings, or to invent equipment – micropipettes and electrode carriers, hand-crafted for techniques that he himself developed and that are today cited in reference books. “Today it’s rarer, but in my time it was something fantastic,” says the researcher, in his calm voice and slight German accent. Head Professor in the Department of Physiology and Biophysics at the Institute of Biomedical Sciences (ICB), at the University of São Paulo (USP), Malnic was born in Milan, the son of Austrians, and moved to Brazil when he was 4 years old; he took Brazilian nationality when he was 23. Based in Sao Paulo, where his father, a chemist, worked as a representative of a German dye company and later for the Matarazzo empire, Malnic studied at the Visconde de Porto Seguro school. He wanted to be a chemist, but his father re recommended the medical school at USP, which he entered in 1952.
He became interested in physiology in the second year of college and, under the influence of Professor Alberto Carvalho da Silva, he specialized in renal physiology, an area at that time little explored in Brazil. He did post-doctoral studies at Tulane University, in New Orleans, between 1961 and 1962 and at Cornell University Medical College, New York, between 1962 and 1964, in the laboratory of Gerhard Giebisch, with whom he is still collaborating today. Back at USP, he set up the laboratory of micropuncture and renal microperfusion, a method of collecting fluid from renal tubules (where water, minerals and vitamins are returned to the blood, leaving the urine) using micropipettes, he studied the transport mechanisms of potassium in the kidney, as well as the role of sodium, hormonal factors, changes in the acid-base balance and nephrotoxic drugs in the reabsorption of bicarbonate. He has written and published more than 120 papers in international journals, as well as a book on renal physiology. He has already tutored eight Masters and two dozen PhDs and has become one of the main points of reference in his field in Brazil.
In parallel with his laboratory work, he has presided over scientific bodies, like the Brazilian Society of Biophysics (1983-1985), Brazilian Society of Physiology (1985-1988), the Federation of Societies for Experimental Biology (FESBE) and the Institute for Advanced Studies (IEA ) at USP. Now retired, he has witnessed the departure from the scene of some of his disciples, but he continues working actively in his laboratory at the ICB. “We’re already on our third thematic project. We received a lot of money and, in part, this merit is mine, because I’m the coordinator. So that’s why I think that they’ll not kick me out, despite my age,” he says, jokingly. “Right now I only have one PhD student, but I’m trying to get some more. Young people prefer to stick with the youngest ones, too,” he says. Married to teacher and translator, Margot Petry Malnic, the father of two daughters, biochemist Betina and singer and song-arranger, Beatriz, and the grandfather of three girls, Gerhard Malnic gave Pesquisa Fapesp the following interview:
You were born in Italy but you’re Austrian. How did you end up in Brazil?
I was born in Italy, but by accident. My father was born in 1901 in Austria, in what is now Croatia. My surname, Malnic, is Slavic. At home, we spoke German. My father’s family was Slovenian and my mother was Austrian, so I’m descended more from German than Slovenians. My father studied chemistry, graduated around 1925, at a very difficult time for Austria, which was practically finished after the First World War. He worked in several countries after the war, including Germany, Poland and Italy, where my sister and I were born. However, I came here when I was four.
Why did your family come to Brazil?
My father was a chemist, a dye-maker. He worked in Italy, in the textile industry. In Germany, dyes were chemically based. He was hired by a German chemical company to come to Brazil. One of the reasons was that he had had experience in Latin countries. So, he came to Brazil as a representative of this firm locally. We went first to Rio and he soon saw that what was really interesting in terms of industry was São Paulo. We moved here and I went to the Visconde de Porto Seguro School when I was six or seven.
Which was a school for the German community…
It has been for the German community since the 19th century. This school was nationalized, you might say, because of the war. I learned German there, too. At home, I learned a little Latin and Greek, because my father thought it was important. He had learned this at the Gymnasium, in Vienna, so he thought that I had to learn a little. However, the fact is that, when he arrived here he worked for some time at this firm, which soon after was confiscated by the government when Brazil entered the Second World War. That’s when he started working for Matarazzo, in the Belenzinho district, in a textile plant. At Visconde de Porto Seguro, I was a good student. The war didn’t affect me. My father was a person who had resources. He soon got back on his feet and then he bought a farm in the north of Paraná and worked for other chemical companies. I always liked chemistry because my father was a chemist; he even set up a small laboratory at home. However, his suggestion to me was as follows: “Look, here in Brazil the best university is that of medicine; it’s best for you to study medicine.” I followed his advice.
What was the Medical School at USP like when you studied there?
That was from 1952. It wasn’ t very different from today. I came first in the entrance exams. I started studying physiology in the 2nd year of medicine. My professor was Alberto Carvalho da Silva, who would become the director of FAPESP. He was a fantastic person, who set up a good laboratory. He liked me as a student and extended an invitation to me at the end of my 2nd year: “Come and work in the laboratory.” I went and began working there. He worked with vitamins, but I thought that I shouldn’t work with them because there was already someone doing this. Something he wanted to do was to work with the renal part of vitamins, as they were eliminated. We sat down together at the laboratory bench to read a book by American physiologist, Homer Smith; that was in 1954. I started studying the renal excretion of vitamin B1, or thiamin, in dogs. What was new was both studying the renal excretion of vitamins, as well as working with dogs that were not anesthetized. Because there was an idea that anesthetized animals, principally in the case of vitamins, could suffer changes in functioning, we worked with a dog that was not anesthetized. We were careful not to cause pain…It was a bit complicated, because we had to introduce a catheter, facilitate the collection of the urine, take blood samples and, what’s more, the vitamin doses were very complicated. This work led to my first paper, with Dr. Alberto, in the American Journal of Physiology, one of the most prestigious physiology journals.
Were there groups working on this in Brazil?
There were people in the clinical area; Dr. José de Barros Magaldi, for example. He was the head clinician at the Medical School. There was also the group at the Escola Paulista de Medicina [Paulista Medical School], also in the clinical part. However, in physiology there were some people who worked as clinicians and had a laboratory, like Marcelo Marcondes, who is retired now, and Antonino Rocha, who came a bit later; he was a student of Marcondes and we worked together later. Antonino Rocha was killed by someone trying to steal his car; those kind of things happened at that time.
EDUARDO CESARThe fact that you chose renal physiology was influenced by Professor Alberto, is that right?
Yes. We went together to an international physiology congress in Buenos Aires and we talked to Professor Robert Franklin Pitts, who was one of the greatest renal physiologists of the time. He said: “I have two young people who are beginning to work with me and they speak German and are just beginning in a new area of physiology,” which was micropunction. In that book by Homer Smith he described all the first part of micropunction, how to puncture renal tubules and obtain samples. It’s a technique that began at the end of the 1920s and the beginning of the 1930s. They used this technique to get details of the functioning of the renal tubules and also the renal glomerules [functional units of the kidneys that filter the blood]. Professor Pitts said: “Come to my laboratory, which is in New York, at the Cornell University Medical College.” So I got a scholarship from the Rockefeller Foundation. At the time, FAPESP didn’t exist, but the Rockefeller Foundation helped Brazil a lot. It had helped Dr. Alberto himself. He went to New York, but first I had to do a year of basic studies, because there were no post-graduate studies here. I was always a person who accepted good suggestions. I thought the idea was interesting. I spent a year at Tulane University, in New Orleans, studying English, mathematics and biochemistry and I did some physiology courses; I learned a lot from them. My English was good; at home, I had had private lessons. I even knew a lot more math than other people did. So I did a physics-chemistry course, which the others didn’t do, which for me was excellent, because from early on I’d been interested in the mechanisms of ionic transport, for which mathematics was important. So, I managed to complete this course, which helped me a lot on the rest of the internship.
When was this?
This was in 1961, 1962. There were several Brazilians and other people from Latin America on this course. It was a Rockefeller foundation program for encouraging Latin American science. I stayed a year in New Orleans, and from there I went to Robert Franklin Pitts’ laboratory and met two young researchers, who were coincidentally Austrian. They didn’t know I was of Austrian descent: “He”s Brazilian; let’s see how he works.”
Who were they?
Gerhard Giebisch was one of them and the other was Erich Windhager, who went to do an internship in Demark. However, Giebisch was a renal physiologist who already had a lot of experience. He worked for some time with a researcher called Phillis Bott, who had worked with A. N. Richards, the pioneer who had developed micropunction. She was working at the Women’s Medical College of Philadelphia, a medical school for women; at the time that kind of thing existed – a school just for women. When I was about to return to Brazil, Giebisch called her, because she was about to retire: “Do you have any equipment there you don’t want?” She said she had and I brought it with me, when I came back here; there were various pieces of equipment with which I could start working. At the time, it wasn’t so complicated in terms of equipment, but I got a microscope from the School of Medicine, which Dr Alberto managed to get hold of for me, and a series of other things – micromanipulators and a chlorate doser from Phillis Bott. Nowadays, when you arrive in a laboratory like this in the United States there are 10 or 15 other post-doctoral researchers and you see your boss from a distance. At that time, there was Giebisch, a technician, and myself so we really did work together, which was very positive when starting out on something new. We developed a method for microdosing sodium and potassium and then we studied potassium excretion mechanisms. We managed to observe how potassium excretion occurred in various tubular segments, in the renal tubules. Formerly, you only got a comparison between blood and urine and we could puncture the various tubules, the proximal tubule, the distal tubule, separately. We managed to discover how potassium was excreted. What we did at the time was something pioneering; all the physiology books carry this, even today.
What was your return to Brazil like?
I came back to set up a laboratory. Despite the initial difficulties, I managed to do quite a lot of things. The first thing was to study the way of handling chlorate, which I only did because I had suitable apparatus. As I didn’t really know what I was going to do, I decided: “Well, let’s do something that I can do with what I’ve got: I’m going to do micropunction and something with chlorate.” I drew off a sample of tubular fluid and made a fairly complicated dosage. Few people did it, because it was too difficult to do. However, I was stubborn. And as a result I managed to push the work forward at the Medical School; I managed to have published an article on the effect of the diuretic furosemide in the transport of chlorates in Nature. I had gotten hold of the material and I invented the project. Afterwards, I constructed this sodium and potassium microdosing equipment here, which was possible because the physiology people had a type of mechanical laboratory; there were people there capable of building things. See this apparatus [shows an old piece of equipment on display in his laboratory]. It has a small flame in the middle and in this small flame, we put a platinum loop; on top of the platinum loop we placed a drop of water and in the water was the solution taken from the renal tubule. To do that here I had help from my first doctoral student, Professor Francisco Lacaz Vieira. He knew a lot about electricity and electronics. He made the equipment for me; he helped build the electronic part, which were two valves, one on each side, one for potassium and the other for sodium, and it had to have 2000 volts – high voltage, at that time. We did that and it worked. I was lucky to have Francisco Lacaz, who retired two years ago. His uncle, Joaquim Lacaz de Moraes had set up a wonderful laboratory. He was a neurophysiologist, but he was also one of those people who built things, electrical things; the whole Lacaz family was very good at electronics. So he set up a laboratory on top of the current school library. In this laboratory, we had a huge space that was not all used. It had a small electrical and electronic machine shop, Lacaz worked a little with his uncle, but when I came back, I was his tutor, because his uncle had retired. Then he began working with me, which was great because he helped me a lot. He’s a person of great ability, very intelligent; he was the head professor here for a long time. I suggested that he measure the pH in renal tubules, and to do this he managed to make an antimony microelectrode. We produced some very interesting things.
Was it necessary to make your own equipment.
Yes, if you wanted to produce anything. And we then produced the work of the mechanisms of hydrogen secretion and the production of acid by the kidneys. I think that of all my work this was the most cited; Lacaz’s and mine. It had over 300 citations. It was a study of the secretion of the hydrogen ion in renal tubules. It was something that was being done – others were doing it but were not using this same method, this same microelectrode. That was very positive. We managed to do this and other work. Later, Lacaz became independent. He went to the United States to spend one or two years there with a famous physiologist in membrane transport, A. K. Solomon.
Was there a mismatch between what you could do there and here?
There was, but at the time, we managed to do a lot in the lab. There was this small workshop that helped a lot. At the time, we had to develop the equipment we needed with our own hands; I always liked doing that. For a long time we had a good machine shop in the department. Unfortunately, our mechanic died a few years ago and he was not replaced, because nowadays physiology has changed; it’s no longer the type of hands-on physiology it used to be. It’s more the physiology of molecular biology, which has equipment that’s always used, like the spectrophotometer, centrifuges, things like that which you buy but don’t make. I really liked using my hands, which was fundamental for renal micropuncture, because we produced gate electrodes and micropipettes; we had to. You spent the whole day building something or another. Today, it’s not at all common.
What other work would you highlight?
There’s our work with potassium and sodium. Professor Giebisch, in the United States, contributed a lot, but I collaborated with him – even today we collaborate; we did a lot together, especially with sodium and potassium. Here, in addition to sodium and potassium, I also worked with hydrogen. We studied the acidification of renal tubules. This was also first done with microelectrodes, which are micro-instruments made in a micro-forge, where we produce a tip filled with antimony – antimony is a metal sensitive to pH. Then we moved from antimony to an ion exchange resin, which is a kind of slightly oily solution that we put into the electrode that contains a substance that is also sensitive to pH. We also worked with a potassium resin – we still do, even today. At the end of last year, I was at Yale University in New Haven with a student of mine, Lucilia Lessa from Fortaleza, and Giebisch always said, “You’re the only one in the world who knows how to do this technique, microperfusion, dosing potassium with microelectrodes”. I’ve done some 130 pieces of work on this, more or less. That’s not much for the United States, but for us here… And it’s good, because our methods are very complicated, difficult to do and as a result, they take a long time.
EDUARDO CESARHow many researchers have you turned out?
I’ve been responsible for tutoring 20 or so PhDs … Some of them continued using this technique. A person who contributed a lot was Professor Margarida de Mello Aires, who has the office next to mine. She did her PhD with me when I first came back, and about two or three years later, she came to work with me. She’s been working with micropuncture and microperfusion since that time, since 1966, more or less. In addition, there are many others. I did some work with Marcelo Marcondes and with Antonino Reyes, with people from the Medical School. Using micropuncture we studied the renal tubules of rats with kidney disease. Lately, I’ve been working a lot with the pH, acid-base balance; in what way the kidney acidifies the urine, which is one of the very important functions of the kidney. I went to the United States to work with transgenic mice in another area. Today, this is done a lot; it’s a very powerful technique. Here it’s more difficult; it’s difficult for us to produce these mice here. However, I went to the laboratory at Yale University, both in 2004 and at the end of last year, and we worked with mice that lack a transporter of ions, in this particular case, of potassium. Another student, Nancy Rebouças, developed the molecular biology part after an internship at Yale, partly at my suggestion. She’s now one of the most important people in Sao Paulo working on molecular biology. Another colleague, Antonio Carlos Cassola, is working on an electrophysiological technique, patch clamp, which also uses microelectrodes, which he learned in the same laboratory as Giebisch and one of his students, Y. Wang. Our whole group has developed from those beginnings. In addition, collaborating with us is Adriana Girardi, a former PhD student of Nancy Rebouças, currently at the Heart Institute, who is working on both the heart and kidneys; she’s another excellent molecular biologist.
What was it like moving from Avenida Dr. Arnaldo to the Cidade Universitária campus?
We moved here in 1972. We were given an area that was no larger than we had in the Medical School. Now, we’re reducing a little because, since I’m retiring and Margarida too, we have to cut down a little, until they decide to throw us out [laughs]. We’re already on our third project supported by FAPESP. We’ve had excellent help, a lot of money, and this is partly my merit because I’m the coordinator.
What’s the focus of your project?
It’s called “Molecular and functional study of ion transporters in membranes.” We’re investigating how renal tubules transport sodium, potassium and hydrogen. It’s a continuation of what we’ve being doing over the years. Nancy is working on the molecular biology part of the project. Adriana herself, from InCor, and the people from our laboratory have been collaborating in this area; she’s published works on molecular biology, especially with their help and a post-doctoral researcher from our lab, a former student of mine and of Nancy, Luciene Carraro-Lacroix, who is now doing post-doctoral studies in Canada.
At what stage is research in physiology in Brazil? How have we evolved?
At the beginning of last century, physiology depended a lot on individual efforts. In Rio, Professor Carlos Chagas began working at the Oswaldo Cruz Institute. At that time, especially in Rio, college physiology was more teaching physiology, and scientific physiology was done in institutions like Manguinhos. Here it was a little different. The Medical School was established in 1913 and it immediately began, at least in part, to do research and teach at the same time. In Rio, physiology research was done at the Carlos Chagas Institute of Biophysics, still today the most important research center in Rio, and here, most was done at the Medical School. We have several people working in the different areas of physiology, such as cardiovascular, neurophysiology and now endocrinology. The neurophysiology area is pretty good, especially after Professor Michael Covian, from Buenos Aires spent some time in Ribeirao Preto. Here we had Professor Cesar Timo-Iaria, who was a student of Covian’s, but both are now dead. Professor Cesar came from Ribeirão Preto here and set up a neurophysiology group that is still very good. Therefore, physiology grew progressively in several areas. However, there are other good physiology groups in Brazil, especially in Ribeirão Preto, Belo Horizonte and Porto Alegre. In Espirito Santo there’s a good cardiology group, too. There really has been fairly good growth. Post-graduate education has helped a lot; it has attracted a lot of people to the area and improved its quality a lot.
Professor Covian came from Argentina in the 1950s; that country was a reference point in physiology. Is it still?
At the time they already had a Nobel Prize winner, Bernardo Houssay. There were very good biochemistry groups, like the one of the also winner of a Nobel Prize, Luis Leloir. Today, it’s difficult to say whether it’s better or worse… I think it tends to be worse rather than better. With post-graduate studies physiology here has grown a lot.
As president of Fesbe, you were one of the leaders of the movement to remove the barriers on scientific experiments with animals. Was this something important?
Yes, because we have powerful opponents, the animal protection societies, and they caused us a lot of difficulties. In fact, in some places vivaria were invaded to release the animals. Finally came the new law from Sergio Arouca, who died before seeing the bill passed, and it was very positive. Now we need an ethics committee for animal experiments to be created in each unit, in each college. This has already existed for humans. Now they’re starting with committees for animals, which is very positive, because it’s clear that we have to treat our animals very well, give them suitable anesthesia and not let them suffer. This is very important. However, you can’t stop using animals in science; that’s also fundamental.
Up to a point there was a positive side to this pressure, in the sense of encouraging the search for alternative methods, wasn’t there?
Yes, that of treating the animals appropriately. We used to use dogs a lot. They caught dogs in the streets and they were then used in research. The Americans use dogs that are specially bred for this, more than we do. But here it’s difficult to do. My PhD thesis on vitamin B1 was done with a dog. Today we use rats more, because rats are bred specially for this, so it’s more pleasant and a lot more controllable. They don’t seem to be so much like a pet as a dog or cat. I remember that Dr. Alberto used a cat for his nutrition experiments, because cats are very good for studies when we need very homogeneous animals. The size of a cat`s skull varies very little, unlike a dog. The skull of cats is much the same among different individuals, and this greatly facilitates neurophysiology studies. Now, cats are animals that are highly protected by society, so today they are hardly used any more. And you can do a lot with a mouse as well. Mice are used a lot today in molecular biology. We did micropuncture with mice. It’s more difficult, it’s a much smaller animal, but it’s the most suitable for producing transgenic animals.
In an article you wrote for a book on higher education, you talk about merit within the university, how the university should change those who are not working and give access to younger researchers. Does this issue worry you?
Up to a point. At USP there are places like the Medical School that function better, but I remember that when I was starting out there were a lot of weak people; people who did not produce scientifically. Now, with post-graduate studies, this has diminished because to get into the Medical School or the ICB or some other good place people have to go do post-graduation and this generally improves the level of the people a lot. However, not everyone who does post-graduate studies remains very productive. There are people whose productivity falls after a few years. And the question is what to do with these people. They often go over to the administration side or teaching. Or, now, they also go into industry. An interesting development … It’s very difficult for a person to be simply thrown out. This leads to a lot of in-fighting. Theoretically, what really should happen is that career progression should depend on merit. This has perhaps improved somewhat, because people used to teach more than they did research. It’s important that people know how to teach well. However, the ideal thing is to both teach and do research, do both things.
In other countries there’s not the stability of tenure in the career of the teacher-researcher we have here.
Maybe that’s a good idea. But I don’t believe that happens because the class defense bodies are very strong. These bodies are powerful. We should have more advantages, including monetary ones, for those who produce more, as happens in the United States. There, the situation is different because it is entities like FAPESP that pay a good part of their salary. So the person who does not produce loses much of their salary and tends to change his or her life, in order to earn money doing something else. Here we don’t have this. It’s very comfortable. You earn the same salary, which is not the worst there is, until the day you die.
But the funding system has developed…
In almost all states we have foundations that support research, the FAPs. The existence of these FAPs allowed teachers who produce to earn more money for research. They’re going to produce even more, because of this financial support from the FAPs and even from the CNPq [National Council for Scientific and Technological Development]. This is something positive and ends up by choosing the best to continue in the universities. Even teachers with higher production can receive grants from the CNPq, and this is already an additional stimulus. The Medical School has something good. They help with the salary from the Medical School Foundation. The staff who are supported end up earning more – and rightly so. It’s a way of supporting those who work more in the laboratory, because in medicine this is essential. Those working full-time there wouldn’t stay full-time if all they received was the basic salary we earn.
Getting back to your career, what are your plans for the future?
I’ll keep on doing research while it’s still possible. I’m 76 years old and I don’t know how long I can continue. I’m in good health. I’ve already had back surgery, the backbone was pinching the nerves in the legs and I managed to get over that well. Obviously I cut down a little. We always end up with less drive than we had in our youth; that’s inevitable. Even so I’m managing to do quite a lot. The contact with young people, for example, is something that’s stimulating. At the moment, I only have one PhD student and an undergraduate scientific research student. Young people prefer to stay with the younger ones too. But we always end up getting someone else. From the viewpoint of research, I’m still working on the excretion of potassium and with transgenic mice. In fact, our ICB import sector has managed to import these mice. However, I’ve also collaborated with colleagues in studies on tubular acidification from the point of view of the transport mechanism of H ions.