Fertility NEWS LETTER

Ideal fertility : ICSI / IVF & Genetic Center India

Vol IV Issue 12, December 2007

In This issue

  1. Aging of cells, role of Telomeres
  2. Calibrating the CO2 incubator
  3. Approach to genetic problems in Rural setup.by Dr.Shantanu Abhyankar
  4. Training in IVF and Embryology

In Previous Issue

  1. What is an Embryo –Current Perspectives
  2. Collection of oocytes in In-vitro Fertilization

Dear Colleges
Hello

At the outset I wish you all the best for forthcoming New year

In this issue we got a write-up from our friend from Wai Satara. I am very happy to put his topic in this news letter but unfortunately I have to do some editing because of constrain of space.

Aging of cells make us mortal and abnormality in aging makes the neoplastic changes in cells. Telomeres are one of most important component of a chromosome so that it protects it form decaying. It is same as our shoe laces are protected from flaying by two plastic covering. As that covering goes off ,the life of shoe lace shortens and one day it breaks off. Controlled manipulations of telomeres may help us one day to control cell aging and treat neoplasia.

Other topic is Calibration of CO2 incubator. One of the most difficult work for embryologist .I tried to make few things clear ,it may be a incomplete effort.

With best wishes ones again

Sincerely yours
Dr. D’Pankar Banerji

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1. Aging of Cells, role of Telomeres

Ordinarily a cell is restricted to about 50 –70 mitotic cell divisons.After reaching this number ,the cell typically becomes senescent and can not continue to divide ( Hayflick Limit).

What Are Telomeres?

Telomeres cap the ends of chromosomal DNA and consist of repeated sequences of the thymine, adenine, and guanine. They are longest at conception and become shorter each time a cell divides.DNA polynerase cannot replicate the tips of chromosomes. This shortening eventually becomes critical and leads to cellular senescence and cell death as important sequences of DNA become exposed or fuse end-to-end. Cell function such as protein production, repair, and signaling, decline as.

Telomere Shortening

In general, aging cells become progressively less able to form and maintain tissue. This dysfunction plays a key role in a variety of presently incurable such as macular degeneration, arteriosclerosis, arteriosclerosis, osteoporosis, skin atrophy, progeria and others. It can also be seen in symptoms we often label as, such as wrinkled and deteriorating skin and the decline of vital organs.

Tumor cells overcomes this shortening by activation agene that encodes telomerase , a reverse transcriptase that replaces the telomeric segments that are normally lost during cell division.

Controlled activation of telomerase may one be of help to control cell aging and preventing the growth of cancer.

2. Calibrating the CO2 Incubator

CO2 incubator is one of the most important equipment in the embryology lab. It is like a surrogate fallopian tube or uterus which gives shelter to early embryos. For that the environment of human fallopian tube has to be mimicked to a maximum extent.

Embryo in the fallopian tube is not exposed to ambient air, hence under less oxygen and more CO2 ,but it is not intravascular too.

Embryos are kept in bicarbonate buffered media and fresh media is towards basic pH. We create an environment by counterbalancing the bicarbonate by increasing the atmospheric CO2 partial pressure so that there is an interaction between bicarbonate buffer and atmospheric CO2 ,and the pH of media is kept constantly towards 7.2-7.4.( CO2 of atmosphere dissolves into the media and creates an acid and balances with bicarbonate)

How to maintain the pH of media in incubator?

  1. Set CO2 concentration as mentioned by the manufacturer of the media and do the culture, in a notion that pH of the media will be adequate .
  2. Measure the pH of media inside the incubator and set the CO2 accordingly .so that pH remains between 7.25-7.35 . It may differ from the recommendation from manufacturer side .

For the first condition :

It is very important to verify that what CO2 level is displayed on the door of incubator is actually the concentration of CO2 inside. There are two methods to measure the CO2 level inside the closed incubator.

  1. By Pyrite equipment : It is an instrument that contains KOH ,and the CO2 from the outlet of incubator is pumped in 18 times ,then absorbed CO2 increases the fluid level in the scale fixed that instrument. The error rate is high (1-1.5 %) and frothing of solution hampers the exact reading. But instrument is cheaper.
  2. By infrared sensor : It is costly, but it is more precise, but has to be certified, that it itself is in good condition, periodically.

For the second condition :

A good pH meter is needed. The pH - meter should be cleaned before being calibrated with pH 7 and 10 standards at the same temp as the media. Thereafter quickly measure the media, same temp as the standards, the pH will vary with temp. The best option is to do blood-gas analysis of hydrogen ions, more stable and more accurate.

3. Approach To Genetic Problems In A Rural Setup

Dr. Shantanu Abhyankar M.D. (Ob & Gy) Modern Clinic,
Wai (Satara) Maharashtra, INDIA Mb 98220 10349Introduction

Introduction

Though India is being looked upon as a rapidly urbanizing society it is still a long way to go before the medical services available to the urbanites are as readily available to the rural masses. Rural practitioners find themselves in a very peculiar predicament. They are faced with a demand for the best out come with minimum expenditure. Financial considerations often overrule necessity. Medical insurance is non-existent and companies that do offer medical insurance, anyway exclude pregnancy and related disorders. Illiteracy, especially amongst the women, blind faith, social taboos, the low social status of women, ill conceived notions about pregnancy, birth defects and untoward out come all contribute to a rural practitioner’s woes.

In rural areas, Doctors are (still) next to god. They are expected to provide clear unambiguous directions as to what to do next and ‘yes’ or ‘no’ answers to all queries. All the talk about counseling in a non-judgmental and non-directional manner comes a cropper when faced with such a clientele.

This not to suggest that it is a lose-lose situation; it is quite the contrary. With a little bit of insight in the subject, a rural practitioner can offer his expertise and services to the optimum.

What Doctors can do?

Be net Savvy.
Doctors need to net savvy. This is the least that they are expected to do. With internet,access readily available, getting on the net is as easy as breathing. Sites such as.

http://www.nlm.nih.gov/medlineplus/geneticbirthdefects.html
http://www.cdc.gov
http://www.birthdefects.co.uk
http://www.marchofdimes.com/pnhec/4439_1206.asp

Provide a wealth of information
Promote Rubella vaccination
Promote preconception folic acid

Folic acid prophylaxis can be offered to prevent neural tube defects. A dose of 400 to 500 mcg daily is suggested for at least three months before conception

Picking up Thalassemia

Thalassemia is common in certain communities in India. Consanguinity will add to the problem .Ideally, a person needs to undergo Hb & CBC with blood indices, hemoglobin pattern analysis and HbA2 determination by HPCL method for a diagnosis as normal, trait or thalassemia major. An RBC count of over 4.5million/cmm, with a MCV of <72fl and MCH of <22 pg is indicative of some type of hemoglobinopathy.

Correct interpretation of TORCH tests

There are misconceptions galore as far as these tests are concerned. Certain guidelines need to borne in mind. Firstly, none of the TORCH agents is responsible for recurrent pregnancy loss. Secondly if ever we have to rationally diagnose and treat TORCH we should be running the test, on paired sera, drawn at least three weeks apart. Testing just once is useless, not informative enough and a waste of money. While a raised or rising IgM (regardless of the quantum) will suggest active infection, which beckons attention; a raised IgG suggests that the woman is presently immune to infection by that particular agent. A rising IgG, where levels rise by over 4 times in those paired tests, will indicate rekindling of disease and will need to be tackled. For all the agents maternal infection does not mean fetal affection. TORCH is not a routine screening test and in absence of any stigmata in the fetus, the results will throw up more questions than answers.

Get recognized as a genetic center.
Follow up problems to the hilt
Consanguinity

Conclusion

Though faced with many hurdles the rural practitioner can contribute to reducing the burden of birth defects in the society. With the advent of internet, the gap between a village and a city is narrowing. The rural practitioner thus should keep up with the times.

4. Training in IVF and Embryology

Module I : Ovulation induction and Intra Uterine Insemination ( One day )
Module II : Conventional IVF and fundamentals of Embryology( Two days )
Module III : Intra cytoplasmic sperm injection, Micro manipulation ( Two days)

Course fees :

Module I : Rs.2000.00 ( US$ 50 )
Module II : Rs.20,000.00 ( US$ 500 )
Module III : Rs. 50,000.00 ( US$ 1250 )

For Module I and II : Rs.20,000 ( US$ 500 )
For all the three modules/Module II and III : Rs.55,000.00 ( US$ 1375 )

Payments :
Draft : in the name of Dr. D’Pankar Banerji,payable at Jabalpur

Accommodation :
Participants can be provided accommodation in nearby hotels at an extra cost ,Range is Rs.700-3000 per day
Lunch will be served without an extra cost

Timing :
10.00 am to 5.00 pm

ONLY TWO PARTICIPANTS PER BATCH FOR MODULE II AND III

Dates :
Throughout the year.

Course directors :
Dr. D’Pankar Banerji,IVF specialist and Dr.Mrs.Rinku Banerji,Embryologist and molecular pathologist

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