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Fluorescence in situ hybridisation (FISH)
Characterization of chromosome anomalies, Y microdeletions and cystic fibrosis mutations in male infertility.A significant proportion (60%) of male infertility is due to an underlying genetic cause. The use of intracytoplasmic sperm injection (ICSI) raises a possibility of transgenerational transmission of genetic defects to the offspring, where the father has a genetic abnormality such as Y-chromosome microdeletions. For proper management of these couples, counselling after clinical investigation, semen analysis, karyotype and DNA analysis to rule out Y-chromosome microdeletions were carried out.
At least three distinct non-overlapping regions on the Y chromosome, named AZFa, AZFb and AZFc (for azoospermia factors a, b and c) are critical for germ-cell differentiation. A fourth locus AZFd was suggested afterwards which lies between loci AZFb and AZFc. Y-chromosome microdeletion detection was carried out on 100 males with a history of either azoospermia, severe oligozoospermia, oligozoospermia or oligoasthenoteratozoospermia (OAT). Multiplex PCR was set up using the Promega Kit Version 1.1 for 18 mutations in the azoospermia factor (AZF) a, b, c and d regions. The PCR products were analysed on 2.5% agarose gel by electrophoresis. Y-chromosome microdeletions were observed in 12 out of 100 patients (12%).
We are now using the modified version of Y-chromosome microdeletion detection kit—as Version 2.0 from Promega which includes 20 mutations.
Cytogenetic and molecular analysis of the fragile X syndrome
Fragile X syndrome is a genetic condition mainly leading to mental retardation in males. It is caused by a fragile site at the tip of the long arm of the X chromosome, which is demonstrable on karyotyping using special tissue culture media. However, cytogenetic methods are not very reliable for fragile X detection, compared to molecular techniques. Molecular studies have shown the presence of an increasing number of CGG (DNA pattern) trinucleotide repeats in every generation. A carrier mother may have 60 to 200 repeats and an affected son will have less than 200 repeats in the fragile X mental retardation (FMR1) gene. Southern blotting with radioactive detection and PCR are used for molecular diagnosis. There is hardly any data on fragile X patients from western India. The aim of this study is to try and develop a non-radioactive molecular method for detection of fragile X, using silver staining detection for PCR analysis and chemiluminescence detection for Southern hybridisation. Children from special schools will be studied to identify families at risk of transmitting this condition, and prenatal diagnosis will be offered to prevent the birth of other affected children.
FISH on follicular fluid to detect gonadal sex chromosome mosaicism in women undergoing infertility treatment
Gonadal chromosome mosaicism is difficult to diagnose, as a gonadal biopsy is required. This cannot be used as a routine procedure. In infertile females undergoing in vitro fertilisation (IVF), follicular fluid is obtained during oocyte pick-up. After collecting the oocytes for IVF, the surrounding follicular fluid is generally discarded. We studied the cumulus cells present in follicular fluid by the fluorescence in situ hybridisation (FISH) technique and found that this was a novel way of detecting aneuploidy in the ovarian tissue. We could detect Turner syndrome mosaicism in this tissue in a cytogenetically proven case of Turner syndrome mosaicism undergoing IVF. The percentage of chromosomal mosaicism is known to vary in different tissues. We could demonstrate that follicular fluid cells are an easily available alternate source of cells to detect chromosome mosaicism by FISH in patients undergoing IVF. The study is ongoing to collect more data on follicular fluid cells and compare it with karyotyping from blood and FISH on buccal smears.
In vitro maturation of germinal vesicle oocytes
In an IVF cycle, about 8.5% of retrieved oocytes are immature at the germinal vesicle stage. Complete maturation of oocytes is essential for the developmental competence of embryos. Human oocyte maturation is considered as the reinitiation and completion of the first meiotic division from the germinal vesicle stage (prophase I) to metaphase II and the accompanying cytoplasmic maturation, for fertilisation and early embryonic development. Immature human oocytes obtained from patients undergoing oocyte retrieval can be matured and fertilised in vitro.
In vitro maturation is slowly becoming established as an alternative treatment option to stimulated IVF for women requiring assisted conception. Germinal vesicle stage cumulus-oocyte complexes collected from patients undergoing oocyte retrieval are being cultured in maturation medium with supplements. Those that reach the M2 stage receive the intracytoplasmic sperm injection (ICSI) and cleavage-stage embryos are obtained. The overall maturation, fertilisation and cleavage rates after ICSI of 150 germinal vesicle stage cumulus-oocyte complexes will be determined.
Fluorescence in situ hybridisation (FISH) in slowly cleaving and arrested embryos
Reproductive wastage is a natural phenomenon to eliminate defective embryos. This has been confirmed by the cytogenetic studies on spontaneous abortions, and the studies recently done on arrested embryos. The fluorescence in situ hybridisation (FISH) technique enables rapid detection of common aneuploidies. A contributory cause of IVF failure is a high number of zygotes with missing or extra chromosomes. Individual blastomeres from slowly cleaving and arrested embryos—which are of no use in IVF—are biopsied using a non-contact diode laser beam treated with hypotonic solution. It is then fixed on slides with an effort to remove the cytoplasm and expose the intact nucleus. The nuclei are located under phase contrast and photographed. FISH for common aneuploidies is performed on these single cells. The signals are analysed to detect aneuploid, mosaic and chaotic embryo
Brief write up
Director – Dept. of Assisted Reproduction & Genetics
Responsible for South-East Asia’s first ICSI baby Luv Singh, in 1994.
First PGD pregnancy in India.
First Laser Assisted Hatching & TESA pregnancies in India.
Over 7500 babies born.
Editor-in-Chief, Fertility & Sterility Indian Edition
Editorial Board Member, Fertility & Sterility, U.S.A.
1995: The All India Ratna Shiromani Award for pioneering work in the field of Obstetrics and Gynaecology.
1995: Outstanding Young Indian Award given by the Indian Junior Chamber.
2001: Listed by the National Institutes of Health Registry (USA) for research in Embryonic Stem Cells
2001: Felicitation by Organisation of Pharmaceutical Producers of India for outstanding research in Stem Cells.
2001: The M.D. Adatia Oration Award – Bombay Obstetrics and Gynecology Society Annual Conference.
2002: Kuvadia-Shah-Vora Oration Award on Stem Cell Therapy – Implications & Importance, by The Bombay Medical Association.
2006: Woman of the Year Award - Zonta International Organization.
2006: Zee Astitva Award for Science and Technology.
2007: Hindustan Times Woman of the Year Award
2009: F ICCI FLO Women Achievers Awards 2009 ""Excellence in the field of Medicine”
2012: Women in the Driving Seat Award by Lavasa Women’s Drive for Science and Medicine
2012: L'Oreal Paris Femina Women Awards 2012 for Science and Innovation.
2014: Selected as Woman of the Year 2014 by Limca Book of Records.
2014: ‘All India Survey’ by Indian Express lists Dr. Firuza Parikh in the top 10 Doctors of india.
2015: 100 most Powerful Women of Asia Award 2015.
Society Memberships / Fellowship:
1. The Indian Society of Assisted Reproductive Technology - President – February 2003 – 2006.
2. The American Fertility Society.
3. The Bombay Obstetrics and Gynaecology Society.
4. The Federation of Obstetrics and Gynaecological Societies of India.
5. The Indian Association of Fertility and Sterility.
6. The Indian Academy of Juvenile and Adolescent Gynaecology and Obstetrics.
7. The Association of Medical Women in India.
8. The Indian Association of Gynecological Endoscopists.
9. Indian Society of Human Genetics.
10. Member of the Drafting Committee to outline guidelines for Assisted
Reproductive Technology at the National level (ICMR Project).
1. President – Indian Society for Assisted Reproduction (2003-2006)
2. Member – Monitoring Committee: Developing cells and tissue engineering – Council of Scientific and Industrial Research (CSIR)
3. Recognized guide for MSc and PhD, Mumbai University
4. Editor-in-Chief, Fertility and Sterility Indian Edition
5. Editorial Board: Fertility and Sterility, USA
6. Recognized teacher and guide for DNB, National Board of Examinations (2014).
Her areas of special interests are reproductive endocrinology and recurrent implantation failure. Dr. Sawkar is in-charge of the Surrogacy Program and has contributed in a major way to the success from this program. She also has a strong interst in the immunology of pregnancy.
Dr. Sujatha Sawkar is a versatile consultant in the Department of Assisted Reproduction & Genetics at Jaslok Hospital & Research Centre. She holds a postgraduate degree in Obs & Gyn. From Karnatak Institute of Medical Sciences, Hubli. She has had specialized training in tubal recanalization surageries under the aegis of the FPAI. In addition she has undergone special training in laparoscopic surgery and ultrasonography.
Proficiency awards at school and college levels. Has presented many award wining papers
She is the senior- most member of Dr. Parikh's team. Young and dynamic, she is part of this unique organisation since the last 14 years. After completing her post-graduation in Obstetrics and Gynaecology, she underwent an intensive training in Ultrasound Imaging at Sir H.N.Hospital, Mumbai. Her keen interest in Endoscopy enabled her to obtain a Diploma in Pelvic Endoscopy from the Kiel and Geissen School of Endoscopic Surgery, Germany. She has an in-depth expertise in various aspects of infertility treatment and has given many lectures in conferences .Her clinical skills have contributed to many smiles on the path to parenthood and she says that her greatest reward is to see happiness on the face of childless couples.In her spare time, she enjoys reading academic journals, novels, meeting new people and has a passion for music. She is also an accomplished singer.
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