Interviews of September/October 2017

14.10.2017:

Prof. N. Raghuram: Dean, Dept. of Biotechnology, Guru Gobindh Singh Indraprastha university at Delhi (Dwaraka in Delhi??). Introduction starts with narration about nitrogen and facts like how human activities produce twice as much reactive nitrogen as nature. Prof. Raghuram has been working with Indian nitrogen group on ways to enhance efficiency of nitrogen usage and reduce pollution. Obtained M.Sc in life sciences from Utkal university in 1984. Obtained M.Phil from Pune univ. in 1987. PhD from Jawaharlal Nehru University in 1994. Co-founder of the Indian Nitrogen Group in 2006, facilitates inter-disciplinary, international consultation on research work and policy aspects related to reactive nitrogen and other nutrients in agriculture, industry and environment. As a modern biologist, his research interests include functional biology, biotechnology of nitrogen metabolism in plants and cyanobacteria. Broader interests include health, education, science dissemination and policy. Has received over 1150 citations from over 50 countries for his research publications and articles.

Dr. TVV starts by mentioning a recent report published by Prof. Raghuram that has been making waves. Close to about Rs. 50000 crores out of the huge amount spent by India for nitrogen fertilizers is wasted due to inefficient usage. Why do we need to worry about nitrogen when it is so abundant in the atmosphere? Because nitrogen in air is N triple-bond N which is inert and difficult to break. Most organisms cannot use atmospheric nitrogen. The first chemical process (Aber Bausch?) developed to break N2 involved heating up to 500 degrees Centigrade and high pressure. Similar conditions used in most fertilizer manufacturing processes. Nutrients like nitrogen, phosphorus, potassium need to come from soil for agriculture. Repeated cropping (??) leads to depletion of these soil nutrients. Intercropping with pulses or soil bacteria needed to replenish nitrogen in soil. Dr. TVV gives the analogy of iron-deficient person needing to take special forms of iron, not iron mineral/metal. Even leguminous plants depend on nitrogen-fixing bacteria to get nitrogen they need. These bacteria use "nitrogenase"; these are nitrifying bacteria which convert atmospheric bacteria or certain other unusable nitrogen compounds to plant-usable nitrogen forms. Leguminous plants host some of these bacteria in root nodules - symbiotic nitrogen fixation. Traditionally, farmers have known through observation that intercropping cereals with pulses improves production.

If intercropping with pulses helps replenish soil nitrogen, and if forests abound with trees and plants without needing fertilizer, why do agricultural fields need artificial fertilizer? In an unmanaged environment like forest, a natural system of recycling exists - litter gets recycled and replenishes the soil. But in man-made, artificial ecosystems like agricultural fields, repeated cropping extracts nutrients but doesn't allow for replenishment as the entire crop is removed from the field (leaves not left to decompose?).

Precision agriculture, where tubes are used to supply small, sufficient amounts of water to crops at various times in a day aren't feasible in India due to high cost of investment. Different aspects to the problem. Improve ways you manage crops - using formulations that release nitrogen slowly lead to more efficient nitrogen usage. These formulations are a bit costlier than conventional fertilizers but this is a policy arena that the goverment is working on - improving these alternatives and making them more affordable. Neem-coated urea produces higher yields. Plants utilize ammonia and nitrates more easily than urea. Neem oil coating slows down conversion of urea to other forms, makes fertilizer available for plants for a longer time and allows the fertilizer to be applied at larger intervals. In conventional fertilizers, around 70% of nitrogen goes unutilized/wasted accordingly to global studies. According to Indian Nitrogen group's study, upto 50% of nitrogen fertilizer is wasted in India ! This amounts to a staggering amount of Rs. 50000 crore per annum.

Other aspect is the limited ability of plants to use nitrogen. Biotechnologists and agricultural bitechnologists working on ways to make crops use nitrogen more efficiently. Interestingly, what we think is efficient usage will vary from crop t crop. For cereal crops, we do not want nitrogen to be diverted to generate more leaves than garins. For leafy vegetables, we need more leaves to be produced. For plants grown for flower, we want the nitrogen to be used better to produce flowers.

Nearly 2/3rd of India's nitrogen fertilizers spent on rice and wheat. Because of higher profit margins in cash crop agriculture, farmers in that sector are able to invest in more efficient fertilizers. But in food grain segment, inefficient use is a major problem - reducing usage of fertilizers difficult due to lower margins and concerns around food security. We need to be identify visually and get a scientific grip on the phenotype and genotype of varieties that use nitrogen efficiently, understand the biological properties of these plants, their nitrogen use efficieny. "Making a horse drink more water and grow better as they say"! Improving nitro.

A wider question has arisen globally - nitrates are pollutants if they mix with ground water, ponds, lakes etc. Health hazards due to nitrates in water, and ecological problems like eutrophication. In the last one decade, there has been a tendency to put all blame on fertilizers for nitrate pollution.

Rain water - nitrogen content is about 9 times more than pre-industrial times. A problem like global warming? Is this all due to fertilizers. Indian Nitrogen group studying how much of reactive nitrogen comes from various sources like automobile exhaust, animals, fertilizers etc. Isolated evidences to show that in some forest areas that are not fertilized, there is higher growth where such rain occurs. Levels of nitrate in ground water in many parts of India is double the WHO-prescribed safe limit. But international community blames only fertilizers. Sewage is the second largest growing contributor to reactive nitrogen - spews out 15% or more of reactive nitrogen, compounding annually. If the nitrogen in sewage can be recycled, nitrogen pollution will be reduced. NO2 comes sources including agriculture, but NOx comes only from burning of fossil fuels, especially diesel combustion in automobiles etc. These have to be addressed; difficult to reduce NO2 from agriculture immediately but the other sources can be addressed easily. Synergy between various programmes (Swacch Bharat, rural toilet construction, Namaami Gange) and nitrogen recycling programmes is possible and is important. World is watching India's emission of pollutants due to India's size of economy and population. India now has the capability to assess scientifically where it stands on these pollution and consumption parameters. India has soft power in various sectors, both within and from Indians working abroad and this should be used to get international attention and cooperation on identifying feasible ways to reduce such pollution.

21.10.2017:

Prof. S.V.Eswaran: Regional Centre for Biotechnology, Faridabad, an institution under DBT, engaged in frontier areas research.

Introductory narration says cataract is the cause of 60% of blindness worldwide, that cataract can be delayed and reversed, so surgery is not always necessary as is commonly thought. Has researched lenosterol synthase that delays and reverses catact in dogs, rabbits. Scientists asking whether this can be extended to huma beings. Served as Dean and Head of Chemistry department at St. Stephen's college, Univ of Delhi and taught for 44 years. Was the principal of Deshbandhu college (Univ. of Delhi) for 2 years. PhD from Delhi University and post-doctorate from Max Planck Institute for Biochemistry at Munich. Studied the transparent protein of the human eye lens and is looking for collaborative work on protein complexes. Is famous for discovering a new molecule named Viswamine (Viswamayene ??), named after Prof. M.A. Viswamitra of IISc, a famous x-ray crystallographer. Received awards including Dr. APJ Abdul Kalam lifetime achievement award (2016) and Indian National Science Academy's teacher award. Published over 70 papers and has been granted many patents.

"Viswamayene" has a high molecular weight, 534, composed of around 100 atoms. [Sparrow chirping sound in the background!]. Was researching on green reagents (as they are known now) called aromatic azides. Azide group contains upto 3 nitrogen atoms covalently bound to an aromatic ring. These are called green reagents because on exposure to light or heat, they release nitrogen which is harmless. When heating one such chemical to 130 degrees, a hitherto unknown molecule was formed. Took many years to name it. It is an unusual compound. Formed from a "long lived" singlet nitrine species which did not convert to the more stable triplet. Normally more unstable species are expected to be very transient and change to a more stable form. The life time of once such "long lived" species discovered by him was measured (at Ohio Univ, with whom he published a paper jointly) to be about 700 picoseconds!! Asks "Can you believe it, 700 picoseconds to be called a long-lived species?!". But it was called long-lived because it did not flip to any of the common stable forms. Instead forms a stable intermediate called carbene - formation of carbene from nitrine is rare, though not unknown. Chemical reactions go through sequence of high and low energy states/pathways like travelling through a rock garden having elevated places and valleys. Invariably the final state would be a low energy state. His papers over the past few years have involved a significant amount of computational and theoretical physical chemistry, and the exact amount of energy in each state has been measured.

First hints of this molecule came in 1986-87. But the structure of the molecule was solved only because of help from Prof. M.A.Viswamitra of IISc, a student of Nobel laureate Dorothy Crofy Hofkin. Prof. Dorothy had said that Prof. Viswamitra is a man with "green  fingers" because he could crystallize anything. Prof. Viswamitra was able to crystallize Prof. Eswaran's molcule into a single crystal and identify its structure. Dr. Eswaran named it Viswamayene for 2 reason - since it has many double bonds like aromatic compounds whose names often end with "ene", and also he wanted the name to have something related to India. "Vishwamayi" in Sanskrit meaning present everywhere - atoms and molecules are present everywhere.

How cashew nut shell finds new applications. Prof. Eswaran has worked on cashew nut shell liquid (a.k.a.CNSL). It is a source of low cost phenolic compounds. Cheap phenylene available from this liquid. This liquid is very corrosive and may cause blindness if it comes in contact with eye. Prepared a polymer from the shell extract. Also prepared a chemical that reacts strongly when exposed to light. From a soluble substance becomes a polymeric resin that could be used in printed circuit boards. Photolithography, a.k.a. UV lithography. Microelectronics industry benefits from this chemical.

Cataractogenesis - evolution/formation of cataract. Dr. Balasubramanian has done pioneering work on this. Protein in eye lens is 100% transparent, but under certain conditions like unwanted oxidation (like production of hydrogen peroxide, "believe me") causes cataract. Published a paper in 2015 in the journal, Nature, based on his research on cataract.

An interesting paper from China - Chinese scientists showed that in a family, first 3 generations were safe w.r.t. eye diseases, but in the 4th generation, 3 people were blind. Traced it to gene mutation that causes early-onset cataract. It is a gene that tends to be conserved very much across generations and is found even among dogs. Lanosterol when applied to eyes of dogs and rabbits was shown by those scientists to reverse cataract in those animals. Now, Prof. Eswaran is trying to take this work forward and .

"Grasshoppers don't eat spinach" - because it knows that if it each spinach, it will get multiple heads (like Ravana, says Dr. TVV). Insect-moulting hormones occurs in huge concentration in spinach, and grasshopper known that these chemicals will cause harm to its biology. Spinach does not contain huge amount of iron - a misconception.

"Should we fear chemistry?". Chemists have been at the receiving end; chemistry has a bad press. All known drugs, esp. in allopathic are synthesized artificially using chemical reactions. Already, 2 Nobel prizes have been awarded for the discovery of carbon-based nano molecules which are believed to have immense medicinal value though they are yet to come into practical use! Very small amounts (about 0.0002 mg dissolved in water) of one such substance has been found to cure Alzheimers and Parkinsons disease - used with approval in Ukraine.

In India though, chemistry is viewed with doubt or fear by public and press. This needs to change. Dr. TVV: Chemical research is interesting - not only leads to discovery of interesting new molecules but also finds practical applications like reversing cataract.

28.10.2017:

Dr. Anirban Basu: Senior scientist, National Brain Research Centre, Manesar, Haryana. Working hard to find a cure to Japanese encephalitis. PhD in immunology from CSIR's Indian Institute of Chemical Biology, Kolkata. Post-doctorate in neuro-immunology from the Neural and Behavioral Science Department of Pennsylvania State University College of Medicine. Joined NBRC's faculty in 2004. Research work focuses on curing diseases of nervous system, currently researching on diseases caused by neurotrophic viruses. His work on minocycline came to limelight in 2008 - it is an antibiotic that is used to treat pimples, and Dr. AB's work showed it could be used to treat Japanese encephalitis. Use of minocycline for treating Japanese encephalitis is in phase 3 trial now, at St. George Medical University at Lucknow. Recently ICMR declared that minocycline could be used for observational study on patients with acute encephalitis symptoms.

Dr. TVV starts interview with the recent encephalitis outbreak tragedy in Gorakhpur. Dr. Anirban says that tropical countries are more susceptible to diseases like encephalitis. Problem with encephalitis, as opposed tro inflammation of other organs like kidney, is that unlike cells of other organs, broken nerves cannot regenerate neurons. Dr. TVV gives analogy of a house that cannot be reconstructed again if foundation is weak.

Why is it called "Japanese" encephalitis? It is no longer prevalent in Japan, but the 1st major outbreak of this disease was reported from Japan in 1871 and hence the name. The recent outbreak in Gorakhpur is not called a Japanese encephalitis but acute encephalitis syndrome, AES, as encephalitis may be caused by other parasites like bacteria. Japanese encephalitis cases have dropped significantly in India in the past few years due to successful vaccination programmes. Says his colleague, Dr. Rashmi Kumar, led the minocycline clinical trials in 2012-2013 for AES. It was shown in animal trials to be a potential cure for encephalitis almost 10 years ago. Dr. Rashmi Kumar published a paper in 2016 based on results from the clinical trial. ICMR has approved it for observational study, not yet a fully-approved protocol drug. If observational study is shown to have the same efficacy as in clinical trials, the drug will be approved.

Japanese encephalitis survivors may show neurlogic, cognitive and psychiatric syndromes. Long-term cognitive impairment even 15 years from the time of infection. Need for social awareness on importance of vaccination. Children more susceptible to encephalitis due to their immune system still developing, but adults may also be affected. Around 137 children died due to JE in Orissa's Malkangiri district in the last major outbreak. Interestingly inn Assam, adult population was more affected, unlike in other states where children were more affected.

Also working on Chandipura virus and West Nile virus, former found only in India. Chandipura virus particularly seen in states like AP, Gujarat. Its affects only paediatric population, and unlike JE virus, its has 100% mortality, so no affected child survives Chandipura virus. Recently, some people have reported dengue virus, Zika virus attacking the brain; this has not been studied well so far and is being researched upon by his institute.

Dr. TVV: NBRC is a unique place that focuses on brain research. Where are we in terms of brain research? Dr. AB: People working here are from multidisciplinary background. Some are focusing on diseases like Huntington's disease, Alzheimer's disease, Parkinsons disease. Some are working on studying brain tumors. Some are working on psychosomatic disorders. Some working on stem cells for treating brain diseases. A few years ago, US started a major initiative for neuroscience research. Neuroscience research is in relatively young worldwide. Planning to start programmes on neuroscience and brain disorders in India. India has a burden of various types of brain diseases - Alzheimer's etc. as well as brain development disorders causes by malnutrition.

Why is brain research difficult? Organs like heart have been studied in modern scientific sense for 100-150 years. Blood transfusion studies started 300 years ago. But brain research has made headway only recently. Because of factors like technology. In the recent years, new technologies have come up that are benefiting brain research. Imaging technology is a sophisticated tool. NBRC has magentic encephalogram that helps pinpoint location where epilepsy originates and thereby helps identify problematic tissue that could be removed to cure the condition. NBRC collaborates with AIIMS, THSTL, RCB. NBRC is important as studying brain disorders requires collaboration between neurologists, psychologists, neurosurgeons, psychiatrists, biologists, and NBRC provides an ideal environment for such collaboration.

Wants to see minocycline go to therapautic use, becomes available off the shelf to clinicians and help save lives, serve the nation. Future questions that NBRC wants to address: Brain inflammation involved in several brain diseases like Alzheimers; inflammation is a critical pathology; wants to study inflammation in general, how it progresses and how to stop it. In diseases like Alzheimers, certain immune cells in brain start releasing proteins out of control and kill normal neurons. Dr. TVV: This is an area that has more questions and answers!

Message to young people: Encourage taking science as a profession. Innovation needed in all aspects of life - agriculture (for food security), health, to address environmental issues.