Tuesday, September 9, 2008

The Great Scientist Dr.Subhas Mukhopadhyay

With tender footsteps Galileo enters the Court room. In the room there were religious priests on the Judge’s chair. At outside, people were assembling in numbers. Here, starts the judgment. The head priest comes forth. Pointing finger toward the man in limelight asks “the center of this universe is the earth. Every planet and star, even sun revolves around it. Do you deny this gospel of the religious scriptures?” Galileo lifts his head and stared at the head priest with a blank look. The head priest asks once again “Do You Admit?” Galileo remained still. All the priests break into a resounding protest “punish him!”, “punish him”.
A curious murmur of a clueless assembly can be heard from the outside. Galileo draws his eyebrows closer, stiffening his jaws bows his head down. As if his weak body can no more keep his head high. Only an indistinct word comes out of his mouth ‘No!’. A celebration of victory irrupts among the priests. An epitome of scientific research, captive in iron fetters, leaves the room amidst the silent assemblers keeping his head down.

The curtain falls on the stage of Academy of Fine Arts. The audience of the filled auditorium could hardly escape the resonance of timelessness of that classical moment. The actors were appreciated for their lively performance by the clapping audience.

Not far in the same city of Kolkata another drama was taking its shape. 18th November 1978. An ‘expert committee’ was appointed by the Government of West Bengal under the medical association to decide over the fate of a convict named Dr. Subhas Mukhopahyay. His charges are, one, he claims to be the architect of a human test tube baby named Durga.(3rd October, 1978). Secondly, he denounced the report to the media before being cleared off by the Government bureaucrats. Thirdly, he made this impossible possible with few general apparatus and a refrigerator in his small southern avenue flat while others cannot even think of it, although, having all the expensive resources in their hand.(in this research Dr. Mukhopadhyay was assisted by Prof. Sunit Mukherjee and Dr. S.K Bhattacharya.). Fourth and most important allegation, he never let his head down by the Government Bureaucrats and his straightforwardness always attracted jealousy out of his peers. The committee was presided over by a radiophysics and it was comprised of a gynecologist, a physiologist and a neurophysiologist. Interestingly, none of them were having any knowledge about modern reproductive technology. The question came from one of the expert “Where did you keep these embryos?” Dr. Mukhopahdhyay said “in sealed ampules.” Then he asked again “How did you seal an ampule?” Surprised Dr. Mukhopadhyay replied “as usual by heating”. From here started a questioning and counter-questioning session, which needs not to be mentioned, was utterly meaningless. “Oh! Embryos do not die of heat while sealing?” there were people who never saw embryos in the entire span of their lifetime!

So, when the justice was predetermined it is needs no mention that everything in that day was stuck in an uncontrolled debate woven carefully by a diplomatic net.
The Committee put forward its final verdict, “Everything that Dr. Mukhopadhyay claims is bogus.”

Only before 67days, means on 25thJuly 1978, 11.45(PM) world’s first human test tube baby Louise Joy Brown was born at Oldham General Hospital in England.The Architects were Robert Edward and Patrick Steptoe. In their procedure they collected Ovum By using Lproscope. At first they observed the evolution and development of the Ovum for a long span of time and then collected it through a small incision. Ovum thus collected is then fertilized by sperm on a small disc. When it forms into an embryo scientists placed it into the womb. But Dr. Mukhopadhyay without using laparoscope collected ovum by undergoing a small operation in the vagina. He increased the number of ovum collected by using a hormone and developed embryo. Lastly, he placed it in the womb. Consequently, the probability of pregnancy increases to higher order.

Thanks to his peers and Government bureaucrats he ultimately handed with a punishment. He had been transferred to ophthalmic department, which sealed his prospect to work on hormones.
In 19th June, 1981 He put a tragic end to his life.

An insulting silence carried on with every passing day. Many of you know that according to the Scientific documents “Harsha”(16th August 1986) become the first human test tube baby of India. The invention went to Dr. T.C Anand Kumar, Director of ICMR(Indian Council of Medical Research). In 1997 he came to Kolkata for participating in Science Congress. It was when all the research documents of Dr. Mukhopadhyay were handed over to him. After meticulously scruitinising and having discussion with Durga’s Parents he became certain that Dr. Mukhopadhyay was the architect of a human test tube baby. This eminent scientist once mentioned in a journal on ‘A critique of Mukerji’s technique’:

“The brief description given by Mukerji in his letter dated 19 October 1978 to the Director of Health Services, Government of West Bengal, the reports he gave over the television interviews and reported in the lay press describe how Mukerji carried out the procedure of in vitro fertilization.

Collection and evaluation of spermatozoa

Durga’s father was found to have a low sperm count, according to Mukerji who obviously was aware of the diagnostics value of semenograms as evidence by the papers he represented much before the WHO laboratory manual for the examination of human semen came out in 1980. Mukerji also knew that such a condition can he effectively treated with gonadotropins. Gonadotropins therapy is now routinely used to treat men with low sperm counts.

Ovarian stimulation

In his report dated 19-10-1978 to the DHS, Mukerji stated that he had treated Mrs Agarwal with hMG 76 ampoules given twice a day an don alternate days and starting from day 3 to day 9 of the cycle. She was given 6000 I.U. of hCG on day 11 of the cycle and she was subjected to oocyte aspiration some 48h later. Mukerji was able to aspirate 5 folicle by this method. In todays context this ovarian protocol will not sound outrageous because controlled ovarian hyper stimulation is the standard procedure for all women subjected to IVF. However, until 1980 december ‘conventional wisdom’ in Britain, Australia and USA dictated that stimulated cycles were unsuitable for oocyte collection and therefore oocyte aspiration was restricted to natural cycles. It was only in 1981 that other scientists resorted to ovarian stimulation. The Australians who were the next to announce the birth of IVS babies, began to use clomiphencitrate for ovarian stimulation in 1981. the Norfolk group in the USA, who were the third to report the occurrence of an IVF baby, began to use with hMC and hCG in their IVF programme with success in 1982. It is noteworthy that Mukerji was far ahead of his time in successfully using an ovarian stimulation protocol before anyone else in the world had thought of doing so.

Ovum pick-up for IVF was a problem during the early days. The British team had used a laparascope to harvest oocytes. The advent of ultrasonography later on opened out a new avenue to aspirate oocytes transvasicaly under ultrasound guidance by making apercutaneous insertion of a long needle traversing the abdominal wall, the urinary bladder and finally the ovarian follicle.

The advent of the transvaginal probe has made it possible to collect oocytes per vaginum and this is the standard procedure used now.

Mukerji’s originality was that he was able to access the ovaries by a very simple operation on the wall of the vagina. Stimulated ovaries enlarge and drop down towards the Pouch of Douglas. A small incision on the posterior wall of the vagina would allow the ovaries to fall through this opening. This reminds one of how a biologist gains rapid access to the ovaries of rats by a very simple lateral, abdominal incision. The entire procedure was accomplished within a couple of minutes. Mukerji’s research experience with rats and humans had helped him take this very simple and original approach of gaining access to ovaries by posterior colpotomy.

One wonders why this very simple approach did not gain popularity. After all today oocytes are aspirated per vaginum under ultrasound guidance.

In vitro culture techniques

The freshly aspirated oocytes were incubated for 4 hours before inseminating them with the husband’s semen that was processed in protein-supplemented Tyrodes solution. This is exactly what is done even to this day in almost all IVF programmes to accomplish in vitro oocyte maturation; processing semen is essential for ‘sperm activation’. The oocytes were exposed to processed semen for a period of 24 hours and later incubated for another 72 hours in a mixture of cervical-uterine fluids. The use of such fluid is not described elsewhere. However, the use of a synthetic fluid, similar to that found in the human fallopian tube, has been described to be useful for in vitro embryo culture procedures.

The methods of in vitro fertilization and embryo growth are described in detail in Mukerji’s letter to the DHS dated 19 October 1978 as well as in a publication in an obscure journal. Mukerji’s stated ‘…It also appears that for cryogenic preservation of embryos with a relatively larger number of blastomeres (more than 8 cells) may be preferable’.

‘Few pre-ovulatory human oocytes collected from a married by surgery were fertilized with spermatozoa from the husbands and cleaved in vitro and subsequently frozen slowly to about 196oC after stepwise treatment with dimethyl sulfoxide. One such frozen embryo was subsequently thawed slowly and when transferred into the uterus of the woman apparently resulted in the production of a clinically normal female baby after normal period of gestation’.

Here is clear published evidence of how exactly Mukerji carried out his version of in vitro fertilization and embryo transfer.

Cryopreservation of embryos from mice, rabbits, sheep goats and was reported between 1971 and 1979. The first report on the successful cryopreservation of four to eight cells human embryos appeared as late as 1981 and Trounson and Mohr reported the first successful clinical outcome of the transfer of thawed human embryos in 1983. A WHO report states ‘embryo cryopreservation has now become a routine adjunct to IVF procedures, and various methods of freezing are employed. The method that has yielded the best results in terms of simplicity, efficiency and reproducibility is one that involves freezing of one to three-day-old embryos (one to eight cells) in a controlled biological chamber that cools the embryos to sub-zero temperatures in the presence of a cryoprotectant 1,2 propanediol. Other cryoprotectants that are used are dimethyl sulfoxide (the same cryoprotectant was used by Mukerji) and glycerol.

It may be noted that Subhas Mukerji reported the successful cryopreservation of a eight cell embryo, storing it for 53 days, thawing and replacing it into the mother’s womb, resulting in a successful and live birth as early as 1978- a full five years before anyone else had done so. This small publication of Mukerji in 1978 clearly shows that Mukerji was on the right line of thinking much before anyone else had demonstrated the successful outcome of a pregnancy following the transfer of a 8-cell frozen-thawed embryo into human subjects transferring 8-cell cryopreserved embryos.” (Current Science, Vol .72. No. 7, 10th april1997)

In Dr. T.C. Anand Kumar’s initiative Dr. Mukhopahdhyay is mentioned as the architect of first human test tube baby in a document related to the subject of artificial intercourse in ICMR India’s first test tube baby “Durga” whose parental name is Kanupriya Agarwal works in a Multinational Company as a Marketing Executive in Delhi. 
On her 25th birthday she first time exposed her identity in a ceremony organize in the memory of Dr. Mukhopahdhyay. She spoke about her creator in front of media and proved once again that her creator’s claim was not bogus.

But the sky is still overcast. There are brave scientists fighting the cause of Dr. Mukhopahdhyay and projecting his genius to the people. The worthy names to be mentioned in this regard are like Dr. Sumit Mukherjee and Dr. T. C. Anand Kumar. In the ‘Dictionary of Medical Biography’ (Volume 4: M-R) edited by W. F. Bynum , Helen Bynum,enlists names of 1100 Medical Scientists from100 countries around the world for the path breaking contribution to the medical science. Only three names found there place in that dictionary from the city of Kolkata. The names are; Sir Ronald Ross, Shri.U.N.Brahmachari and Dr.Subhas Mukhopadhyay. What is more ridiculous is that after his death, one by one three scientists Howard Jones, Gleichar and Tronson (Australia) claimed three of their separate researches as their own which was, in fact, done by Dr. Mukhopadhyay well before them. One of these scientists even found his research published in famous Journal “Nature”. Sometimes we cannot help asking ourselves “How long should we wait for receiving pats from other countries for recognizing our work and validate it by a letter of appreciation.” Should we judge a scientist by the colour of his skin and not by their work?

Before reading this piece in public I altered its contents several times. May be, sometimes, I thought it was my over ambition to put a sea in a bowl. It is because I am still not in a position, in respect to wisdom as well as age, to understand and explain Dr. Mukhopadhyay’s work. At the beginning of my writing I couldn’t control myself being attacking to see such injustice with a man of Dr. Mukhopadhyay’s stature. But Prof. Sunit Mukherjee repetitively reminded me that I am a student of science. If I say something about Dr. Mukhopadhyay I should only emphasize on his work because, the identity of a truly devoted scientist is his work. May be, he never wanted to associate his pains with his inventions.

Still, my emotions paved its own way I entered in a different world without my knowledge. I have seen Giordano Bruno is burned alive. Leibniz work is imitated. The Hungarian scientist János Bolyai who left his family and friends to prove Euclidean geometry, his epitaph reads “this man wasted his life in worthless work”. Prof. Jagadish Chandra Bose was once maligned by holding responsible for stealing apparatus from laboratory. In the history of science the list for deprivation is long. But the people who never give up to unfavorable situation proved their character as true devotees to their work. But in future we may not be able to see such characters around us any more.

My friends, if my effort is successful to present even a microscopic portion of Dr. Mukhopadhyay’s work to you I will think that I have paid my respect to this great scientist of our country.

Dr.Subhas Mukherjee (1931- 1981)

1931 , 16th January – Born in Hazaribag , Bihar, India

1955 – Obtained M.B.B.S. from National Medical College, Calcutta. Stood First in Gynecology and obtained also Hemangini Scholarship. Obtained B.Sc.(Hons.)in Physiology from Calcutta University.

1958 – Ph.D. From Calcutta University in Reproductive Physiology under the stewardship of Prof. Sachchidananda Benerjee.

1960 – Marriage with Namita and left for U.K. under Colombo Plan to study Endocrine Physiology.

1967 – Ph.D. From Edinburgh in Reproductive Endocrinology, Develop method for estimation of hormone (LH).He worked at the Clinical Endocrinology Research Institute with Prof. John A. Loraine and obtained his Ph.D. In his research work a new method of assay of LH was found out. In those days no reliable method was available. The technique depends on ovarian cholesterol depletion of intact immature rats pretreated with PMSG & HCG. He also worked at the Royal Infirmary, Edinburgh.

1967- 1975 – Lecturer, Reader and Professor of Physiology , NRS Medical College, Calcutta. Fundamental research on the origin of HCG, which was postulated by Theodore Langhans as far back in 1870. He contradicted the view and established that it is of deciduas origin rather then trophoblastic in origin. Hence he had to ply with the embryos and early stage foetus, which helped him in creating Test Tube Babies. Fundamental work on the use of Testosterone in the management of female infertility. Outstanding hypotheses that stress is related to female infertility, which was presented at the International Conference on physiological sciences held in Paris in July 1977. Later workers are now confirming such relations.

1978, 17th August – Invited by the Rotary club of Belur to speak on Test Tube Baby.

1978, 3rd October – Birth of India’s first test tube baby was announced. His method was different from that followed by Steptoe & Edwards and it was also the world’s first test tube baby from frozen and thawed embryo. He did not undervalue the Indian Science and published his paper in Indian Journal of Cryogenics and presented his findings in Indian Congress , in January 1979.

He discussed his experiment in the “ Fifth International Congress on Hormonal Steroids 1978” held at New Delhi in November – December 1978. He also discussed the matter with Professor John Biggers of the Laboratory of Human Reproduction, Harvard Medical School, UAS and Prof. B.B.Sexena Professor of Bio-Chemistry and Endocrinology of the Cornell University Medical School in the USA and Professor Kenneth Ryan Professor of gynecology of Havard. He was invited by the Benaras Hindu University to speak on the subject of embryo transfer. He also talked on the subject at Gauhati Medical College at the conference of the Gauhati Obstetrical and Gynecological Society, where he was presented with a “Manpatra” or a scroll of honor.

Dr.Mukherjee’s research was highly acclaimed in National & International forums as evidenced by the views of the different scientists.

“ As a biologist and gynecologist together , development of adequate culture media, after analyzing the tubal fluid seem to be not a problem for him. Then he succeeded in freezing human blastocyst long before any one else in the world had done so. His successful invitro fertilization and embryo transfer of a frozen blastocyst on another cycle of women which leads to pregnancy and delivery of a healthy baby in 1978, in the same year when Louis Brown was born, Should finally enter into scientific literature.

The concept of transfer of one or two blstocyst after deep friezing & then synchronizing hormonal cycle remain so far the untold method to increase the pregnancy rate up to a high level then physiologically possible. We have to admire that the early idea of Subhas Mukherjee will be a millstone in the future of routine procedure in invitro fibrillation.”

- Prof. Kurt Semm
Kiel University, West Germany
Feb. 8, 1984

“Purpose to conclude by speculating on some of the possible social and ethical implications that bioengineering cryogenics working together can present in gating the better of human infertility which, I fear, may grow into a sizable problem in the not too distant future. The genious of Dr. Subhas Mukherjee lay in that he dared and seized upon a problem which belongs to the future and that a not-to-near future. No woner that his peers, who perceived a major threat to their complacency, set upon and destroyed the man”

- Prof. Ashok Mitra
I.C.S. June 19, 1985

“ It is indeed the dedication, hard work & perseverance of late Dr. Subhas Mukherjee which allowed him to obtain the first test tube baby in India. Like anything new in life people always criticize people always doubt. But truth always remains.

The work invitro fertilization has now just started flourishing in India, but we lost lot of time in our bitter internal conflicts & rivalries. It is flourishing very well outside. But that is what I call as a bane of our won Indian culture & mentally in scientific fields. We sometime enjoy turning down ourselves more then that is necessary. This is not to mean that one should not interact, criticize, debate on each other. We talk about developing a new scientific culture but unless & until we learn a bitter lesson, we won’t understand how much valuable time we lost in this important discovery made by Late Dr.Mukherjee. I think we will still continue to long behind in many fields including our own which relates to human development.

The work of Late Dr. Subhas Mukherjee was related to promotion of fertility which is equally necessary when we talk about family planning.”

- Dr. Badri N. Saxena
Sr. Deputy Director General , Indian Council of Medical Research
New Delhi , Jan. 16, 1987

“ The above mentioned thoughts aptly describe the trials and tribulations of a young, creative scientist, Late Dr. Mukherjee who left a legacy of rich thought , sincerely of purpose and devotion to work against all odds. No one else but who has undergone similar experiences can deeply understand how Dr. Mukherjee might have felt when new concepts attached views which were of non-belief and sometimes even ridicule”

- Dr. Anil R. Sheth
Institute for Research in Reproduction (ICMR)
Bombay, Jan 16 1988

1978, December – Govt. of West Bengal set up an Enquiry Committee which did not acknowledge his claim. As no physiological or biochemical techniques including DNA fingerprinting can distinguished between in-vivo and in-vitro fertilized normal baby, Dr.Subhas Mukherjee categorically stated to all concerned people that he cannot provide a sure proof for his work on IVF and ET. But he also asserted that if there was a genetic marker, though a surrogate, then proof will be obviously established. Even after these long years, as far is known, no country in the world has accepted a legal connotation of a test tube baby on the municipal birth certificate. Some professional bodies also heckled him in some meetings. He was denied passport to go to Japan where he was invited to speak on IVF & ET. He was later transferred to the Eye dept. of Medical College, Calcutta in June 1981.

1980- He had an heart attack and was transferred to RG Kar Medical College, Calcutta.

1981 19th JUNE - He had a tragic death.

My special thanks to ---

1. Prof. Sunit Mukherjee
[Formerly professor and Head Food Technology and Biochemical Engineering dept.(J.U)]
Secretary—Dr.Subhas Mukherjee Memorial Reproductive Biology Research Center
2. Architect Of India’s First Test Tube Baby- Dr.Subhas Mukherjee
Edited by- Prof. Sunit Mukherjee & Dr. S.C.Lodh
3. Kuntal Bhattacharya ( Human Resource Executive )
4. Mr. Surajit Goon And Mr. T. R. Panigrahi ( Lecturer B.C.D.A Collage of Pharmacy & Technology )
5. Times News Network 15 October 2003
More Information Log On to - www.drsubhasmukherjee.com