The story of an IISc scientist who brought molecular techniques into research on the deadly disease in India
A young T Ramakrishnan, who lived with his family in Ernakulam in Kerala, helplessly watched his mother being sent to Andhra Pradesh’s Madanapalle sanatorium, one of the few such medical facilities in the country created to treat people suffering from long-term illnesses. His mother was battling the highly infectious disease, tuberculosis (TB).
This was in the early 1930s when TB had no cure. Though TB has been around for over thousands of years, it was only in 1882 that Robert Koch, a German physician and microbiologist, identified the bacterium responsible for the disease in humans – Mycobacterium tuberculosis.
With abysmal treatment options, TB was one of the main causes of death worldwide, especially from the end of the 19th century to the beginning of the 20th century. The only option at that time was to admit TB patients to a sanatorium, where they would receive ample rest, nutritious food and calcium supplements. But Ramakrishnan’s mother’s immune system was not strong enough to fend off the disease — she never returned home.
“This loss may have drawn him to the field of TB research,” M Venugopalan, Ramakrishnan’s brother tells me as we sit at IISc’s Archives and Publications Cell office.
Ramakrishnan would go on to spearhead TB research in the country. He introduced molecular biology at the Institute at a time when researchers were reluctant to embrace this field at IISc.
“This loss may have drawn him to the field of TB research,” M Venugopalan, Ramakrishnan’s brother tells me as we sit at IISc’s Archives and Publications Cell office
Born in 1922, Ramakrishnan excelled in academics: he was a triple gold medallist, standing first in the entire Madras Presidency. But growing up he had serious health concerns ̶ he would get bouts of migraine that continued into adulthood. This worried Ramakrishnan’s father, K Karunakaran Nayar, the Principal at Maharaja’s College in Eranakulam. On his insistence, Ramakrishnan stayed home for two years after his graduation, says Venugopalan.
But soon things changed for the better. One day, Ramakrishnan’s stepmother happened to notice something in an old newspaper that was used to wrap groceries. It was an advertisement from IISc, inviting applications for master’s programme. She encouraged Ramakrishnan to apply.
This was a turning point in Ramakrishnan’s life, Venugopalan adds with a smile. After Ramakrishnan graduated in biochemistry from IISc, he was sent by the Government of India under the International Science Exchange Programme to the University of British Columbia (UBC), Canada, to pursue his PhD. He studied metabolism in microbes, paving the way for his future research on TB.
In 1957, when Ramakrishnan returned to his alma mater as a lecturer in the Pharmacology lab [now called the Department of Microbiology and Cell Biology], the situation of TB in India was grave. To control TB, in 1951, the government had launched a campaign of mass vaccinating people with Bacillus Calmette-Guerin (BCG). This vaccine was a breakthrough that came out in 1908, when researchers, Albert Calmette and Camille Guérin from the Pasteur Institute in Paris, found that weakened bacteria could provide protection against TB. And in 1921, after relentless years of research and the First World War, they tested this on a young boy in Paris. The vaccine contained a weakened Mycobacterium bovis, which causes TB in cows. It works by provoking the immune system to attack the bacteria, preventing a possible TB infection in the future.
Though it continues to be used to this day, its impact on preventing transmission is limited. Ramakrishnan pointed this out in his 1999 review, while also stating that several countries have been successful in controlling infectious diseases, including TB, by improving nutrition and adopting public health measures such as better sanitation, safe drinking water, and so on.
Besides control, they were other means of fighting TB. In 1943, researchers made a discovery that would change the way that TB is treated. From soil samples collected at the Rutgers Agriculture School, USA, researchers, Albert Schatz and Selman A Waksman, isolated the bacterium, Streptomyces griseus, which produces the antibiotic streptomycin. This antibiotic proved effective against many bacterial diseases including TB. However, not long after, some strains of M. tuberculosis began showing resistance to this antibiotic, making it a huge threat.
In the years that followed, many scientists across the world began zooming into genes of bacteria to understand how they cause disease. This was around the time that the structure of DNA had been revealed by James Watson and Francis Crick. Biologists were beginning to understand the link between DNA, RNA and proteins — how the information in DNA makes proteins. This was the beginning of the era of molecular biology.
But scientists at IISc still used the classical and tedious approaches of biochemistry: they would isolate a protein, purify it and analyse the enzyme kinetics.
“People didn’t appreciate that molecular biology could provide any insights or valuable information. We can say that he [Ramakrishnan] embraced novel approaches,” explains MS Shaila, Ramakrishnan’s PhD student and now, Emeritus Professor at MCB.
This change didn’t happen overnight. In 1961, four years after he joined IISc, Ramakrishnan met Jacques Monod, a pioneer in the field of molecular biology, at the Society of Biological Chemists in Delhi. Monod encouraged him to carry out molecular biology studies on M. tuberculosis, according to Shaila. Since molecular biology hadn’t yet found its footing in India, in 1962, Ramakrishnan spent two years at EA Adelberg’s lab in Yale University to train himself in this field.
Back in his lab at IISc, Ramakrishnan now combined techniques from both molecular biology and biochemistry to understand the differences in metabolism between harmful and harmless strains of Mycobacterium. He showed that the harmful strains showed an increase in levels of isocitrate lyase enzyme with age, while the harmless strain showed no such increase. Mycobacterium depends on this enzyme for its survival. Humans lack this enzyme, making it an ideal drug target. Researchers are now trying to design drugs that can inhibit this enzyme.
Ramakrishnan was also interested in understanding how antibiotics work against the TB-causing bacterium, and how they developed resistance to the antibiotic. In a study published in 1973, Shaila and he showed that streptomycin acts against TB by blocking protein synthesis in Mycobacterium.
In another study, Ramakrishnan along with another PhD student, CV Sunder Raj, discovered a virus called I3 mycobacteriophage from a soil sample at IISc. The virus was shown to attack the bacterium, Mycobacterium smegmatis, which is related to Mycobacterium tuberculosis. These viruses integrate their genome into the bacterial genome. He published this discovery in the journal Nature. “Phages are genetic tools to map the chromosomes of the bacteria. He isolated and characterised this phage”, explains Shaila.
Ramakrishnan was also interested in understanding how antibiotics work against the TB-causing bacterium, and how they developed resistance to the antibiotic
Ramakrishnan wrote an account of his work titled The Agony and Ecstasy of One of Midnight’s Scientists, in which, he says that experiments in modern biology require fairly expensive chemicals and instruments, which were not easy to come by in those days, as they were not supported by Indian agencies. Thanks to international grants he received from Rockefeller Foundation, Wellcome Trusts and Nuffield foundation, he could carry out sophisticated experiments. “Through one such grant from the Wellcome Trust”, says Shaila, “he could import a whole lot of radiochemicals”. She adds that though he kept them in his lab, students from other labs were also given access to them.
Ramakrishnan also established IISc’s first facility to study viruses with the help of WE Levinson from the University of California, San Francisco, USA, who spent a year at the Institute. Levinson studied Rous sarcoma virus, known to cause cancer. They together showed that the anti-TB drug, Isoniazid and their metal derivatives inhibit the growth of this virus. They found similar results with other viruses as well.
From then on, he set out to address other diseases that India was grappling with at that time. He studied the virus Rinderpest that caused major outbreaks, killing thousands of cattle, goats and pigs. The virus is now eradicated. He also investigated rotavirus that causes extensive diarrhoea in children. Collaborating with the Vani Vilas hospital, Bangalore, Ramakrishnan and team screened about 500 children in Bangalore for the presence of rotavirus and to determine its type as part of a WHO project. Their results helped WHO compile a list of the types of rotavirus, a crucial first step in the fight against them.
Ramakrishnan also was instrumental in setting up the plant tissue culture laboratory at IISc with the help of CS Vaidyanathan from the Department of Biochemistry. Ramakrishnan then hired Lakshmi Sita, who went on to work on developing disease-resistant sandalwood trees and other transgenic crops in her lab.
“He [Ramakrishnan] had the vision to do something which was taken up by others in the country, maybe five or 10 years later”, says Shaila.
Ramakrishnan retired in 1982 as the Chair of his Department. He was then made senior scientist in a DST project, where he collaborated with Shaila to study rinderpest. In his INSA account, he says that he used this opportunity to tie up the loose ends of TB projects he was involved in.
Remembering his association with Ramakrishnan, P Ajithkumar, Professor at MCB, who interacted with him as a student and later as a faculty, says, “Even after his retirement, he used to stop by my lab and talk to me about the latest developments in tuberculosis research”.
In recognition of Ramakrishnan’s contributions to TB research, Barry R Bloom, a noted global health scientist, lauded Ramakrishnan’s work in a book he edited titled Tuberculosis, Pathogenesis, Protection and Control.
“He [Ramakrishnan] had the vision to do something which was taken up by others in the country, maybe five or 10 years later”, says Shaila
Ramakrishnan had other interests too. Venugopal remembers him as a voracious reader. He had huge stacks of books, which he catalogued. Venugopalan calls him a living encyclopaedia as he would often quote people like Shakespeare and Bernard Shaw. “He also loved music, gardening, travelling and photography,” says Venugopalan, as he shows me Ramakrishnan’s photo album, in which photos are meticulously organised, with neat labels for each photograph describing the occasion and the people captured in them.
For Venugopal, what stood out about Ramakrishnan was that he remained a student throughout his life. When Ramakrishnan felt the need to have a strong grounding in mathematics, he enrolled to attend lectures of VG Tikekar from the Mathematics Department, along with his students. “He loved meddling with gadgets”, recalls Venugopalan. “Often, I used to see his bed strewn around with different tools. Occasionally, he used to blow up fuses!”
Ramakrishnan was also conscious of his civic responsibilities. Venugopalan says Ramakrishnan was appalled at the poor civic, law and order situation due to the apathy of the concerned authorities in Sharda Colony in Basaveshwara Nagar, where his family settled. “Women feared going out due to antisocial elements. The concerned authorities turned a deaf ear despite repeated complaints. So he formed a resident welfare association, Citizens for Civic amenities, Basaveshwara Nagar, despite facing several bureaucratic hurdles.” This association, Venugopalan says, “filed a Public Interest Litigation against Bangalore Development Authority and Corporation, followed by a Contempt of Court Petition in the High Court of Karnataka for their callous negligence in discharging their duties.”
According to Venugopalan, the association won both the cases and things dramatically began to improve. Sharda Colony is now one of the best residential localities in Bangalore, Venugopalan claims.
Even before he retired from IISc, Ramakrishnan was diagnosed with Parkinson’s disease. But he didn’t let this come in the way of his work and led an active life: he drove his Ambassador car until the age of 79, went for regular walks. His wife, Parvathy Devi was a strong pillar of support, Venugopal says. After losing his wife to breast cancer, in a few months, Ramakrishnan’s Parkinson’s symptoms grew worse: food began entering his windpipe, leading to pneumonia.
“He was looking forward to being a part of IISc’s centenary celebration later that year. But he succumbed to pneumonia and Parkinson’s at the Fortis Hospital, Bannerghatta Road, just before completing 86 years in February 2008,” recalls Venugopalan.