ICMR BULLETIN VOL.19-No.-7-JULY-1989.pdf
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ISSN 0377-4910
NITY HEALTH cell
ICMR
BULLETIN
Bga
VoL19 No.7
July, 1989
ELISA FOR THE DETECTION OF CELL MEMBRANE
ANTIGENS OF SPERMATOZOA
It has been suggested that one of the causes of female
infertility may be an antisperm antibody* and that con
ception may not take place if the iso-antibodies are pres
ent in high concentrations2. It has also been suggested
that sperm agglutinating activity may develop as a result
of obstruction in efferent sperm transportation pathways.
Spermatozoa from men with sperm agglutinating activity
in the serum often show pronounced agglutination and
exhibit reduced cervical mucus penetration. Studies have
demonstrated that sperm agglutinating activity detected
in both male and female is caused by sperm agglutinating
antibodies3. There is therefore a need to develop simple,
quick and sensitive methods for the detection of antiWperm antibodies to help in the management of infertility.
The presence of antibodies to spermatozoa in unex
plained infertility3 and following vasectomy4 can be
detected by microsperm immobilization1, passive haem
agglutination2, microsperm agglutination3 and micro
cytotoxicity tests5. The assays for detecting anti H-Y
antibody include microcytotoxicity6, hybrid antibody
method7 and haemagglutination test8. These conven
tional methods are time consuming, have an element of
subjectivity and require fresh semen samples with good
count and motility. An enzyme linked immunosorbent
assay (ELISA) which is considered to be a quick and
sensitive method was developed in the laboratory of the
Institute for Research in Reproduction (IRR), Bombay,
to detect murine anti H-Y antibody and for screening
mouse monoclonal antibodies directed to human sperma
tozoa. Phylogenetic conservation of H-Y antigen across
the species has been reported9. Cross reactivity of this
antigen using human and mouse spermatozoa was shown
by IRR scientists by microcytotoxicity assay and
ELISA10. It is for this reason that human spermatozoa
were used as target cells to detect the activity of murine
H-Y antibody in ELISA.
Preparation of H-Y Antibody
Anti H-Y antibody was raised in syngeneic Swiss
mice by immunizing young females (8-10 wk old) intra
peritoneally with male splenocytes (1X107) in PBS with
complete Freunds adjuvant—CFA (1:10) four times, at
weekly intervals11. The ascites fluid was tapped and the
activity of H-Y antibodies studied by cytotoxicity assay
and ELISA and specificity by absorption with male and
female spleen cells10.
Preparation of Monoclonal Antibodies Directed to
Human Spermatozoa
Washed human spermatozoa obtained from normal
healthy volunteers were used for immunization. BalbC
mice were immunized with 1X106 washed spermatozoa in
Division of Publication & Information, ICMR, New Delhi - 110 029
0.1 ml of PBS mixed with equal volume of CFA at fort
nightly intervals. After four immunization the splenocytes of mouse which had a titer >1:3200 was used for
fusion. Fusion was carried out using mouse myeloma
cells SP2/0 using polyethylene glycol 1540 (J.T. Baker
Chemical Co., Philipsburg, NJ). Supernatants were
screened for the presence of antibody using ELISA12*13.
Preparation of Mouse Epididymal Sperm Samples
Epididymis from 3-4 mice were teased in PBS, pH
7.2 to release spermatozoa. The tissues were removed and
sperm suspension was centrifuged at R.T. at 400 g for 20'.
The cells were washed thrice and the cell count was
adjusted to lX107/ml. Fifty microliters of this sperm sus
pension was added per well.
Preparation of Human Spermatozoa Samples
Semen samples with good count (>68X106/ml) and
motility (>50%) and negligible number of contaminating
cells and without agglutination were used for these stu
dies. Freshly collected semen samples were liquified at
37° C for 30 min and centrifuged at 400 g. The cells were
washed thrice with PBS, pH 7.2 and used for coating
plastic plates. Cell membrane antigens of spermatozoa
were solubilized with 3 M KC114. The protein concentra
tion of solubilized antigen was determined by the method
of Lowry and coworkers15 and used for ELISA.
Plastic Plates
Polyvinyl (Dynatech) and polystyrene plates from
different sources (Greiner, Linbro, Falcon, Flow) were
tested for their suitability for the assay.
Influence of Assay Conditions
Optimal conditions for the assay were determined by
pretreatment of plates with PHA, fixing cellular antigens
with glutaraldehyde and by the addition of BSA and
Tween 20 to washing buffer (PBS, pH 7.2).
Procedure for ELISA
In experiments where phytohaemagglutinin (PHA)
was used for pre-coating, 10 jug of PHA in 100 ul of PBS
was added per well and incubated for Ih. The solution
was taken out and wells were washed thrice. Washed
mouse epididymal spermatozoa or washed human ejacu
lated spermatozoa were used for coating the plates. Fifty
microliters of sperm suspension containing 5X105 sper
matozoa was added per well. The uniform layer of sperm
suspension was allowed to dry at room temperature.
These cells were fixed with 100 ul of 0.025 per cent gluta
raldehyde for 30. Plates were flicked to remove glutaral
dehyde and washed four times, with PBS containing 0.05
74
per cent Tween 20 (polyoxyethylene sorbitant). Hundred
microliters of appropriately diluted H-Y antibodies or
100 ul of hybridoma culture supernatant was added per
well and incubated for Ih, the supernatants were
removed and the wells washed thrice. Rabbit anti mouse
immunoglobulin raised, purified and conjugated to horse
radish peroxidase at the I RR, Bombay by the method of
Nakane and Kawaoi16 was used for this assay. The conju
gate was titrated and a dilution of 1:800 was found suita
ble for the assay. Fifty microliters of diluted conjugate
was added per well and incubated for Ih in dark at R.T.
and the wells were washed four times. The substrate was
prepared by mixing 4.86 ml of 0. IM citrate buffer,
5.14 ml of 0.2 M NazHPO, buffer, 10 ml of distilled water,
8 mg of orthophenylene diamine and 50 ul of 30 per cent
H2O2. Two hundred microliter of freshly prepared sub
strate was added per well. The reaction was stopped after
15' by the addition of 100 ul of 4 N HSO. and the optical
density was read at 492 nm in a titertek ELISA recorder.
is
The only difference when soluble antigen was used
that 10 ug/ well of antigen was added in carbonatebicarbonate buffer of pH 9.6. The protein concentration
can be reduced significantly when pure antigen is used for
coating. Control ascites fluid and myeloma cell super
natants were used as negative controls. Antiserum of
BalbC mice immunized with spermatozoa was used as a
positive control. When the absorbance of the test sample
was more than twice that of the negative control the test
was considered as positive.
Titration of mouse anti H-Y antibody was carried
out using both mouse epididymal spermatozoa and
washed human ejaculated spermatozoa. The antibody
was found to react with both mouse and human sperma
tozoa (Fig.1) confirming the high cross reactivity of this
antigen. The titre of the antibody in ELISA was 1:160
whereas it was 1:32 by microcytotoxicity assay.
6
The results of titration of H-Y antibody carried out
under different experimental conditions are presented in
Fig.2. Washed human spermatozoa were used as the anti
gen in these studies. Control ascitic fluid was used as a
negative control. It was seen that pretreatment of plates
with PHA had no added advantage. Fixation of cells
with glutaraldehyde and addition of Tween 20 and BSA
to the washing buffer gave better and consistent results.
Microtitre plates of different makes such as polysty
rene (PS; Greiner, Linbro, Falcon, Flow) polyvinyl chlo
ride. (PVC; Dynatech), were compared for their ability to
bind sperm and the results are presented in Fig.3. Mono
clonal antibody (Mab) to sperm antigen (F8) was used
as the antibody, while washed immobilized human sperm
HSP+glutaraldehyde+PBS
o—o
1.2
1.0
0-8
06
20
-40
80
160
Reciprocal Dilution
Fig.l Titration of H-Y antibody using mouse epididymal spermato
zoa and hunan ejaculated spermatozoa. Values represent differences
between absorbance values (492 nm) for H-Y antibody and control
ascites fluid. Bach value is mean of triplicate values.
was used as the antigen. The PVC plates gave high back
ground values as compared to PS plates. It was seen that
reactions in the PS plates yielded higher absorbance
values as compared to reactions in the PVC plates.
Amongst the PS plates Linbro plates gave higher absor
bance values. The other three brands of PS plates viz
Galcon, Flow and Greiner were found suitable for this
assay (Fig.3). The difference in these results was only
marginal indicating that all brands of PS plates could be
successfully employed for this assay.
J________i____
20
*40
. I________ Li
80
100
Reciprocal Dilution
Flg.2 Titration of H-Y antibody under different experimental condi
tions. Human spermatozoa (HSP) was used as the antigen. Values
represent differences between absorbance values (492 nm) for H-Y
antibody and control ascites fluid. Each value is mean of triplicate
values.
method of choice for screening antibodies directed to a
wide variety of antigens.
Ever since the first report of Engvall and Perlmann17
and Van Weemen and Schuurs18, ELISA has been
adapted and extensively used for various purposes and
many assay methods have"been developed. Compared to
commonly used antisperm antibody assayb3’19 ELISA
technique has a number of advantages especially to
screen a large number of hybridoma supernatants.
Fig.4 shows the comparison of absorbance values
when the antigen is presented in different forms, namely
cellular and soluble. For this study 3 Mabs to human
sperm were used. The immobilized cells (cellular antigen)
gave higher readings with all the three Mabs tested. Mye
loma supernatant from SP2/0 cell was used as the
control.
ELISA does not need excellent donor semen in
terms of count and motility. It has been noted by scien
tists of I RR that coated microlitre plates could be stored
at 4°C for several months without any loss of quality.
The assays commonly used for screening hybridoma
supernatant are RIA and ELISA. It is known that some
of the monoclonals do not form suitable antigen anti
body complexes as they are directed to a single epitope.
Therefore, over the past few years ELISA has been the
Sonicated membrane antigens of spermatozoa were
used earlier at the IRR20 for ELISA. A relatively mild
method with 3M KC1 resulted in high recoveries of mem
brane associated HLA antigens and has been shown to be
-superior to sonication14. In the present studies 3M KC1
75
Reciprocal Dilution
Fig.4 Comparison of cellular and soluble antigens of human sperm
using 3 Mabs. Values represent differences between mean absorbance
values (492 nm) for Mab and myeloma supernatant. Each value is
mean of triplicate values.
Fig.3 Suitability of different types of plastic plates for ELISA.
Values represent differences between mean absorbance values
(492 nm) for Mab and myeloma supernatant. Each value is mean of
triplicate values.
was therefore used for solubilizing cell membrane anti
gens of spermatozoa. With soluble antigens the absor
bance observed was low as compared to the cellular
antigen and hence only cellular antigen was used subse
quently. It was observed that precoating of plates with
PHA had no added advantage and it was therefore not
used in subsequent assay. The results obtained at IRR
also indicate that low concentration of glutaraldehyde is
convenient and suitable for fixing spermatozoa since it
achieves adequate fixation and yields minimum back
ground absorbance. Addition of Tween 20 to washing
buffer gave reproducible results and increased the sensi
tivity of the assay and therefore this was used in subse
quent assays. ELISA has been shown to be as sensitive
and reproducible as any other binding assay18 and more
sensitive than sperm agglutinating assay to detect anti
bodies to spermatozoa21.
ELISA was used for the detection of antibodies to
H-Y antigen using mouse epididymal spermatozoa and
human sperm. The reliability of ELISA to detect H-Y
antigen was assessed by comparing it with microcytotoxi
city assay and it was found that the former was more
sensitive10.
These studies demonstrate that ELISA can be suc
cessfully employed for the detection of cell membrane
76
antigens of spermatozoa using monoclonal and alloantibodies. It has also been shown that results may be influ
enced by the conditions of the assay. The ELISA is found
to be more sensitive, and fast as compared to other con
ventional tests and is particularly suitable for screening
large number of hybridoma supernatants. It is suggested
that antibodies directed to spermatozoa and found posi
tive by ELISA should also be assessed by other conven
tional methods.
References:
1.
Isojima, S., Li. T.S. and Ashitaka, Y. Immunologic analysis o
sperm-immobilizing factor found in sera of women with unex
plained sterility. Am J Obstei Gynec 101: 677. 1968.
2.
Rao. S.S. and Rangnekar, K.N. The comparative evaluation of
the haemagglutination and spermagglutination techniques to
detect and assay auto-and iso-antibodies to human spermato
zoa. bu J Fertil 15: 127. 1970.
3.
Friberg, J. Clinical and immunological studies on sperm
agglutinating antibodies in serum and seminal fluid. Acta
Obstei Gynec Scand Suppl 36: 3, 1974.
4.
Hellema. H.W.J. and Rumke, P. Sperm autoantibodies as a
consequence of vasectomy. 1. Within 1 Year postoperation. Clin
Exp Immunol 31: 18, 1978.
5.
Husted, S. and Hjort, T. Sperm antibodies in serum and semi
nal plasma. Int J Fertil 20: 97, 1975.
6.
Goldberg, E.H., Boyse, E.A., Bennett, D., Scheid, M. and Cars
well. E.A. Serological demonstration of H-Y (male) antigen on
mouse sperm. Nature 232: 478, 1971.
7.
Hammcriing, U., Aoki, T.. De Harven, E.. Boyse, E.A. and
Old, LJ. Use of hybrid antibody with anti-yG and anti-ferritin
specificities in locating cell surface antigens by electron micros
copy. J Exp Med 128: 461, 1968.
8.
Furusawa, M.. Kotani, M. and Takeuchi, H. Male specific hae
magglutination in mice. Nature 200: 182, 1963.
9.
Wachtel, S.S., Koo, G.C. and Boyse, E.A. Evolutionary conser
vation of H-Y (male) antigen. Nature 254: 270, 1975.
10.
Iyer, S.V., Nandedkar, T.D. and Hegde, U.C. Production of
H-Y antibody in the ascites fluid of mouse and localization of
the antigen on cells and tissues. Gamete Res 22: 37, 1989.
11.
Tung, A.S. Methods in Enzymology, Vol.93, Eds. H.V.
Vunakis and J.L. Langone. Academic Press, New York. p.12,
1983.
12.
Hegde. U.C., Khole, V.V. and Iyer, S.V. Production and char
acterization of monoclonal antibodies to human sperm specific
antigens. Indian J Exptl Biol 26: 917 1988.
13.
•
•
Kohler. G. and Milstein. C. Continuous cultures of fused cells
secreting antibody of predefined specificity. Nature 256: 495,
1975.
14.
Reisfeld, R.A., Pellegrino. M.A. and Kahan, B.D. Salt extrac
tion of soluble HLA antigens. Science 172: 1134, 1971.
15.
Lowry, O.H., Rosebrough, NJ., Lewis. F.A. and Randall, RJ.
Protein measurement with the folin phenol reagent. J Biol
Chern 193: 265. 1951.
16.
Nakane, P.K. and Kawaoi, A. Peroxidase labelled antibody. A
new method of conjugation. J Histochem Cytochem 22: 1084
1974.
17.
Engvall, E. and Perlmann, P. Enzyme-linked immunosorbent
assay (ELISA). Quantitative assay of immunoglobulin G.
Immunochemistry 8: 871, 1971.
18.
Van Weeman, B.K. and Schuurs A.H.W.M. Immunoassay
using antigen enzyme conjugates FEBS Lett 15: 232. 1971.
. 19.
Kilbrick, S., Belding, D.L. and Merril, B. Methods for the
detection of antibodies against spermatozoa. Fertil Steril 3:
430, 1952.
20.
Rajgopal. R. and Rao. S.S. An enzyme-linked immunosorbent
assay for the detection of antisperm antibodies. Indian J Med
Res 70: 440. 1979.
21.
Wolf. H. and Schill. W.B. A modified enzyme linked immuno
sorbent assay (ELISA) for the detection of antisperm
antibodies. Andrologia 17: 426. 1985.
This write-up was contributed by S. Premchandran. S.V. Iyer. V.V.
Khole and U.C. Hegde. Institute for Research in Reproduction,
Bombay.
ABSTRACTS
Some Projects Completed Recently
Regulation of gene expression during cardiac hyper
trophy.
The present investigation was carried out to examine
the molecular signals triggered during development of
cardiac hypertrophy and the stress mediated alterations
brought about in the gene expression under this
condition.
68,000 and 43,000 which got phosphorylated, were
induced by cytosol obtained from hypertrophic hearts.
The peptide mapping analysis confirmed that these
nuclear envelope proteins were the same which were pres
ent in cytosol and got transferred to the nuclear envelope
after phosphorylation.
The DEAE—cellulose column fractions having
maximum effect on transport of RNA in vitro and on the
enzyme activities were analysed by polyacrylamide gel
electrophoresis (PAGE). The 7th and 8th fractions, hav
ing maximum transport activity as well as maximum
alkaline phosphatase activity and Caz+ dependent
ATPase enzymes showed that 73,000, 68,000 and 43,000
molecular weight proteins were present in greater
amounts as compared to other proteins.
The in vitro transcription experiments revealed that
nuclei obtained from the hypertrophic heart showed
maximum 3H-ATP incorporation into RNA as com
pared to the sham nuclei. Further, the maximum
enhancement of transcription in vitro in isolated sham
nuclei by 40 per cent hypertrophic cytosol suggested that
the factors responsible for such stimulation of RNA syn
thesis may be induced during advanced stages of hyper
trophy. Ammonium sulphate fractionation and DEAEcellulose chromatographic separation of cytosolic com
ponents obtained from hypertrophic hearts helped in
identification of the fractions containing factors(s) having
the maximum stimulatory effect on in vitro transcription
in isolated nuclei obtained from sham operated hearts. A
73,000 molecular weight protein was the most enriched
protein having maximum stimulatory effect.
The phosphorylation studies revealed that the
nuclear envelope proteins of molecular weight 73,000,
The phosphorylation studies of nucleoplasmic and
cytosolic proteins revealed that the 73,000 molecular
Cardiac hypertrophy was induced in experimental
lyslem by means of an operational procedure. The cyto
solic proteins involved in the transport of RNA and stim
ulation of mitochondrial protein synthesis was purified
90 fold (molecular weight approximately 86,000).
77
weight protein was labelled at higher level in the cytosol
obtained from hypertrophic heart and translocated to
nucleoplasm when sham nuclei was incubated with
labelled cytosol obtained from hypertrophic heart.
C. Rajamanickam
Department of Biochemistry
School of Biological Sciences
Madurai Kamaraj University
Madurai.
Publications:
1.
Kamala, J., Mariappan, M., and Rajamanickam, C. Changes
associated with rat heart chromatin during cardiac hyper
trophy. J Biochem Intern 13: 271, 1986.
2.
Kamala, J., Mariappan, M. and Rajamanickam, C. Mitochon
dria functions during experimentally induced cardiac hyper
trophy. J Biosci 10: 293, 1986.
Studies on perspectives of circulating immune complexes.
Serum samples from patients with bronchial asthma,
myocardial infarction, systemic lupus erythematosus and
acute glomerulonephritis were analysed for the presence of
circulating immune complexes (CICs), to gain an insight
into the clinical significance, origin, nature and biological
functions of these moities.
CICs could be detected in 30 per cent of the 165
bronchial asthma patients. Of the 165 samples from
patients phenotyped for C3 19 FS, 143 SS and 3 rare
variants could be detected. Similarly, of the 327 samples
from unrelated normal adults, in addition to common C3
phenotypes, 16 rare variants could be identified. Fre
quency of C3f gene was found to be 4.0.
CICs could be detected in 74 per cent of the 125
patients with myocardial infarction. When the patients
were categorized in four groups representing the initial,
acute, post, and recovery phases of infarction, the CICs
could be detected in 70, 80, 56 and 90 per cent of the
patients respectively. Levels of C3 persisted in the normal
range with higher marginal values in most acute phase
patients.
Lipoproteins had a modulatory role on complement
system, which is an integral part of the immune com
plexes. High and low density lipoprotein (HDL and
LDL) activated the alternate pathway of the complement
system in patients with myocardial infarction and thus
could play a role in the pathogenesis of myocardial
infarction/or evolution of infarct size. However, for the
same lipoprotein concentration, degrees of activation by
LDL was much higher than the activation caused by
HDL. Very low density lipoprotein (VLDL) did not have
any effect on C3 activation. CICs isolated from patients
of myocardial infarction triggered spontaneous aggrega
tion of normal human platelets.
Fourteen out of 17 patients of actue glomerulonephri
tis showed the presence of CICs. A higher level of CICs
was found in sera of patients with systemic lupus erythematosus (558 ug of AHG equivalent/ml) and with acut@
glomerulonephritis (787 ug of AHG equivalent/ml).
Glomerulonephritis patients showed a reduced number
of C3b (CRI) receptor in the erythrocytes as compared to
normal subjects. This is of high significance as CRI recep
tor is known to be involved in the clearance of immune
complexes from the circulation and hence its estimation
could be of diagnostic and prognostic importance.
L.M. Srivastava
Department of Biochemistry
All India Institute of Medical
Sciences
New Delhi.
Publications:
1.
Srivastava, N.. Gupta, S.P. and Srivastava, L.M. Association
between C3 complement types and bronchial asthma. Hum
Hered 35: 263. 1985.
2.
Das, N., Boro, N.S., Panchamoorthy, G. and Srivastava, L.M.
Circulating immune complexes in myocardial infarction. Indian
J Med Res 86: 74. 1987.
ICMR NEWS
Meetings of the following Scientific Advisory Com
mittees/Project Advisory Committee/Project Review
Committee of the Council were held during the month:
Meetings of the Scientific Advisory Committees (SACs):
SAC of the National Institute of
Nutrition, Hyderabad
July 3. 1989
(at Hyderabad)
SAC of the Laboratory Animal
Information Service Centre,
Hyderabad
July 4, 1989
(at Hyderabad)
78
SAC of the Food and Drug Toxico
logy Research Centre, Hyderabad
July 4. 1989
(at Hyderabad)
SAC of the Institute of Cytology and
Preventive Oncology. New Delhi
July 17. 1989
(at New Delhi)
SAC of the Centre for Advanced
Research on Endomyocardial
Fibrosis, Trivandrum
July 20-21. 1989
(at Trivandrum)
SAC of the Institute of Immuno
haematology, Bombay
July 21, 1989
(at Bombay)
SAC of the Institute of Pathology,
New Delhi
July 26, 1989
(at New Delhi)
meeting on Review of Ongoing Studies on Non-A NonB Hepatitis, at New Delhi (July 10-13, 1989).
SAC of the Institute for Research
in Medical Statistics (Delhi
and Madras Chapters)
July 31, 1989
(at Madras)
Dr. B.N. Saxena, Sr. Deputy Director-General,
ICMR, participated in the meeting of the Indo-US Joint
Working Group on the DBT-USAID Collaborative
Project on Contraceptive Development and Research in
Immunology, at Washingaton D.C. (July 11-14, 1989).
Meetings of the Project Review Committee (PRC)/Pro
ject Advisory Committee (PAC):
PRC on Biochemistry and Cellular
and Molecular Biology
July 11, 1989
(at New Delhi)
PAC of the Assessment of the
Anti-tobacco Community
Education Programme
July 24-25, 1989
(at New Delhi)
Participation of ICMR Scientists in Scientific Events:
Dr. K. Banerjee, Director, National Institute of
Virology, Pune, participated in the WHO Intercountry
Dr. G.V. Satyavati, Sr. Deputy Director-General,
ICMR, participated in the IV World Conference on
Clinical Pharmacology and Therapeutics, at Mannheim/
Heidelberg, W. Germany (July 23-28, 1989).
Dr. Kamala Krishnaswamy, Deputy Director,
National Institute of Nutrition, Hyderabad, participated
in the Symposium on Drugs and Nutrition, held during
XIX International Congress of Paediatrics, at Paris (July
24-28, 1989).
ICMR AIDED SYMPOSIA/SEMINARS/WORKSHOPS/COURSES/CONFERENCES
Symposium/Seminar/ Workshop/
Course/Conference
Date & Place
Contact Address
International Workshop on Women and
Development: A Critical Appraisal of House
hold Level Research Methodologies.
July 2-11. 1989:
Baroda
Dr. Amita Verma. Child Development Depart
ment. Faculty of Home Science. M.S. University
of Baroda. Baroda.
1 National Workshop on Enzyme Histo
chemistry and immunohistochemistry.
July 5-22. 1989;
Madras
Dr. M. Madhavan. Professor & Head. Department
of Pathology. Dr. ALM Postgraduate Institute of
Basic Medical Sciences. Madras.
Continuing Medical Education on Nuclear
Medicine in Today’s Practice.
July 15-16, 1989:
Hyderabad
Dr. Kakarla Subbarao. Director. The Nizam’s
Institute of Medical Sciences. Hyderabad.
Symposium on Haemolytic Anaemias.
July 30, 1989:
Hyderabad
Dr. G. Madhusudan Reddy. Medical Officer. Insti
tute of Genetics and Hospital for Genetic Diseases.
Hyderabad.
National Conference on Recent Trends in
Physical Chemistry.
August 2-3, 1989;
Calcutta
Dr. R.V. Gopala Rao. Department of Chemistry.
Jadavpur University, Calcutta.
Continuing Medical Education Programme
on Rheumatology Update.
August 12-13, 1989;
Hyderabad
Dr. Kakarla Subbarao. Director. The Nizam’s
Institute of Medical Sciences. Hyderabad.
VII IMA/CGP National Conference on
General Practice.
August 18-20. 1989;
Varanasi
Dr. S.P. Gupta. Organising Secretary 7th IMA/
CGP National Conference. IMA House. 7/31.
Chetganj, Varanasi.
National Seminar on Strategies for Optimal
Management of Cancer in Developing
Countries.
August 24-27. 1989:
Srinagar
Dr. Shad S. Akhter, Department of Medical Onco
logy. Sher-I-Kashmir Institute of Medical Sciences,
Srinagar.
National Seminar on Newer Challenges in
Medicine.
September 9-10, 1989:
Jabalpur
Dr. B.N. Shrivastava, Professor and Head, Depart
ment of Medicine and Dean, Medical College,
Jabalpur.
Contact Address
Date & Place
Sy mposium/ Semina r/ Workshop/
Course/Conference
Seminar on New Trends in Methodology for
Social and Population Research.
October 1-3. 1989;
Tirupati
Prof. K. Mahadevan, Department of Population
Studies, S.V. University, Tirupati.
National Workshop on Burns Injuries and
their Sequele.
October 7-8, 1989;
Chandigarh
Dr. C.P. Sawhney, Professor & Head. Department
of Plastic Surgery, Postgraduate Institute of Medi
cal Education and Research, Chandigarh.
XVI Annual Conference of Indian Immuno
logy Society and Symposium on Tropical
Immunology.
October 12-14, 1989;
Sevagram
Dr. B.C. Harinath, Professor & Head. Department
of Biochemistry, Mahatma Gandhi Institute of
Medical Sciences. Sevagram. Ward ha.
XXII Annual Convention of the Indian
Dietetic Association.
October 19-20, 1989;
Hyderabad
Dr. Indira Ray. Reader. Home & Social Science
Department. Calcutta University. Calcutta.
Brachytherapy Workshop for Cancer Cervix.
November, 1989;
Betul (M.P.)
Dr. Anand Vijay Kumar. Secretary. Paramedical
Section. Christian Medical Association of India.
Smruti Theatre Compound. Nagpur.
All India Symposium on Biophysics of Cell
Structure and Function.
November 6-8. 1989:
Chandigarh
Dr. P.C. Mangai. Chairman. Department of Biophysics. Panjab University. Chandigarh.
Seminar on Zoonoses.
November 9-10. 1989;
Dibrugarh
Dr. L.P. Datta. Director. Regional Medical @
Research Centre (ICMR). P.B. No. 105, Dibru- ”
garh.
All India Symposium on Reproductive Bio
logy and General Endocrinology.
November 26-28. 1989:
Baroda
Prof. P.M. Ambedkar. Department of Zoology.
M.S. University of Baroda. Baroda.
Workshop on Recent Advances in Intensive
Care.
November 26-30. 1989;
New Delhi
Dr. H.K. Kaul. Professor & Head, Department of
Anaesthesiology. All India Institute of Medical
Sciences. New Delhi.
XXXIII All India Obstetric and Gynaecolo
gical Congress.
December 28-30. 1989;
Patna
Dr. J. Mishra, Department of Obstetrics and
Gynaecology. Hospital for Women. Patna Medical
College Hospital. Patna.
Seminar on Indian Speech. Language and
Hearing Tests-the proposed IS HA battery.
February 8-9. 1990;
New Delhi
Dr. S.K. Kacker. Professor & Head, Department
of Otorhinolaryngology. All India Institute of
Medical Sciences. New Delhi.
COUNCIL’S TRAINING PROGRAMMES
Leprosy
Endocrinology
At the Central Jalma Institute for Leprosy, Agra:
At the National Institute of Nutrition, Hyderabad:
Training Course on Leprosy for Medical Officers
(August/September, 1989).
Annual Training Course on Endocrinological Tech
niques and their Application (August/September
1989).
Virology
At the National Institute of Virology, Pune:
Short-term Course on Detection and Isolation of
Arboviruses Using Mosquito Inoculation and Mos
quito Cell Culture (October, 1989).
80
Nutrition
At the National Institute of Nutrition, Hyderabad:
...
Annual Certificate Course in Nutrition (December
1, 1989 to February 28, 1990).
Occupational Health
Immunohaematology for Technicians (August 131, 1989).
At the National Institute of Occupational Health,
Ahmedabad:
Training Course in Blood Banking Technology and
Immunohaematology for Medical Officers (August
1 to September 30, 1989).
Refresher Course for Industrial/ESIS Medical
Officers (October 9-14, 1989).
Training Course on Disaster Management of
Hazarduous Chemicals in Industry for Industrial
Physicians, Chemists, Safety Officers, Hygienists
(November 6-10, 1989).
Laboratory Animal Technology
At the Laboratory Animal Information Service Centre,
National Institute of Nutrition, Hyderabad:
Cytology
At the Institute of Cytology and Preventive Oncology
New Delhi:
Indo-US Workshop on Chemoprevention of the
Uterine Cervical Dysplasia with Nutrient Inter
vention (December, 1989).
aematology
Training Course for Laboratory Animal Super
visors (September 1 to November 30, 1989).
Clinical Pharmacology
At the Seth G.S. Medical College, Bombay:
At the Institute of Immunohaematology, Bombay:
Training Course in Blood Banking Technology and
Training Course in Clinical Pharmacology (Novem
ber December, 1989).
SOME AVAILABLE ICMR PUBLICATIONS
Price
Rs.
Nutrient Composition of Indian Foods (1989). by B.S. Narasinga Rao. K.C. Pant and Y.G. Deosthale
15.00
Growth & Physical Development of Indian Infants and Children (1972). Reprinted 1989
10.00
Studies on Weaning & Supplementary Foods (1974). Reprinted 1984
6.00
Studies on Pre-School Children (1974). Reprinted 1984
6.00
A Manual of Nutrition (1973). Second Edition 1974. Reprinted 1984
2.50
Low Cost Nutritious Supplements (1972). Second Edition 1975. Reprinted 1977
1.50
Menus for Low Cost Balanced Diets and School Lunch Programmes Suitable for South India
(1959). Third Edition 1977, Reprinted 1984
2.00
Menus for Low Cost Balanced Diets and School Lunch Programmes Suitable for North India
(1959), Third Edition 1977, Reprinted 1984
2.00
Some Common Indian Recipes and their Nutritive Value (1964), Fourth Edition 1977.
Reprinted 1984. by Swaran Pasricha and L.M. Rebello
6.00
Nutrition for Mother & Child (1962). Third Edition 1978. Reprinted 1983, by P.S. Venkatachalam
and L.M. Rebello
5.00
Japanese Encephalitis in India (Revised Edition 1980)
5.00
Some Therapeutic Diets (1969), Fourth Edition 1981. Reprinted 1983. Revised 1988,
by Swaran Pasricha
2.00
Recommended Dietary Intakes for Indians (1981), Reprinted 1984
4.60
Lathyrism: A Preventable Paralysis (Revised Edition 1983)
... HEALTH CEL
COMMULI 11.
326. V Main, 1 Block
Koram-ngala
BangelotC~660034
India
1.50
91
A Manual of Laboratory Techniques (1983). Edited by N. Raghuramulu, K. Madhavan Nair
and S. Kalyanasundaram
Fruits (1983). by Indira Gopalan and M. Mohan Ram
30.00
Count What You Eat (1989). by Swaran Pasricha
The Anophelines of India (Revised Edition 1984). by T. Ramachandra Rao
4.50
150.00
Depressive Disease (1986). by A. Venkoba Rao
Medicinal Plants of India. Vol.2 (1987)
58.00
136.00
2.50
These publications are available on prepayment of cost by cheque or postal order (bank and postal charges will be extra) drawn in favour of
the Director-General. Indian Council of Medical Research. New Delhi. Money Orders are not acceptable. All correspondence in this regard
should be addressed to the Chief. Division of Publication and information. Indian Council of Medical Research. Post Box No.4508. Ansari
Nagar. New Delhi-110029.
TOPICS FOR POSTGRADUATE MEDICAL THESES
In an attempt to improve the quality of research leading to theses submission by Postgrad
uate medical students, the ICMR proposes to compile and disseminate a list of researchable
areas of national and scientific relevance as possible topics for theses. The list will also include
suggestions regarding availability of necessary facilities and expertise for further guidance to
the postgraduate students. In the first instance, the theses topics will cover 3 disciplines viz
Microbiology, Gynaecology & Obstetrics and Community Medicine. It is hoped that these
efforts will help in attracting suitable talent and in sensitization of medical students to biomedi
cal research of high quality. Further, it is also hoped that the effort will also result in creation
of a more conducive en ironment for fostering biomedical research. The Topics identified will
be announced in the forthcoming issues of The ICMR Bulletin.
Indian Council of Medical Research
Awards and Prizes 1989
The last date for submission of nominations/applications for the ICMR Awards and Prizes has been
extended up to September 15, 1989.
EDITORIAL BOARD
Chairman
Members
Dr. A.S. Paintai
Director-General
Dr. Usha K. Luthra
Dr. Badri N. Saxena
Dr. S.P. Tripathy
Chief, Publications
Dr. G.V. Satyavati
Editor
Dr. N. Medappa
Printed and Published by Shri G.P. Kulshreshth for the Indian Council of Medical Research, New Delhi
at the ICMR Offset Press, New Delhi
R.N. 21813/71
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