ICMR BULLETIN VOL. 26-No.-7-JULY-1996
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- ICMR BULLETIN VOL. 26-No.-7-JULY-1996
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ISSN 0377-4910
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BULLETIN
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Vol. 26, No. 7
July, 1996
HIGHLIGHTS OF RESEARCH ON DIARRHOEAL DISEASES
UNDER THE INDIAN COUNCIL OF MEDICAL RESEARCH
The problem of cholera has been with us for centuries.
As a matter of fact, the Indian sub-continent came to be
known as the “home of cholera”. A great deal of work has
been done in India and abroad on various aspects of the
problem of cholera including its control, but even today
there exist many gaps in our knowledge and understanding.
The Indian Council of Medical Research established the
Cholera Research Centre in February, 1962 to conduct indepth studies on cholera and to suggest remedial measures.
The Centre initiated a number of studies in collaboration
with WHO, Geneva including two cholera vaccine field
trials, cholera carrier, serological and chemoprophylaxis
studies and a number of hospital-based clinical trials. The
Centre was recognized as the WHO International Reference
Centre for Vibrio Phage Typing in 1968, and designated as
the WHO Collaborating Centre for Reference and Research
on Vibrios in 1978, for providing referral services to the
member countries. In view of its expanded scope and activi
ties covering a large number of microorganisms in relation
to enteric diseases, the Centre was renamed as the National
Institute of Cholera and Enteric Diseases (NICED) in 1979
It was designated as the WHO Collaborating Centre for
Research and Training in Diarrhoeal Diseases in 1980. In
the recent past, the efforts of the National Institute of Chol
era and Enteric Diseases have been supplemented by the
scientists working at the ICMR Centre for Advanced Re
search in Enteric Diseases, at the Christian Medical College
(CMC) Hospital, Vellore and at the ICMR Advanced Centre
for Enteric Diseases Research and Training at the All India
Insitute of Medical Sciences, New Delhi.
Studies on Cholera
The global cholera scenario experienced an unprec
edented event during 1992-93, when a new toxigenic
non-Ol strain1"6, designated as Vibrio choleras 0139 Ben
gal7, appeared in the Indian subcontinent and later was
reported from many other countries of the region as well as
from developed countries like the UK and USA. Scientists
of the NICED have the credit of bringing this to the notice
ofglobal scientific community for the first time. The NICED
with its expertise in various disciplines immediately em
barked upon research related to V.cholerae 0139 in the
areas of microbiology, clinical medicine and epidemiology.
These studies revealed that from the beginning of November
1992, there was an unusual increase in the relative rate of
isolation of V.cholerae non-01 as compared to V.cholerae
01 from patients of cholera-like illness admitted to the
Infectious Diseases Hospital, Calcutta1. By December 1992,
V.cholerae non-01. became the predominant organism
(>95%) isolated from patients with cholera-like diarrhoea
examined in this hospital13. The unusual trait of these iso
lates is their ability to produce large amounts ofcholera toxin
(CT)1. Simultaneously, similar strains were identified from
different parts of the country3. This was the first report of
a major epidemic of cholera caused by CT producing
V.cholerae non-01. Clinical studies clearly showed that the
Division of Publication & Information, ICMR, New Delhi - 110 029
disease produced by V.chblerae non-01 (0139) was clini
cally indistinguishable from the disease caused by V.cholerae
01 and that the management is essentially the same4,8. It was
earlier demonstrated by a well controlled clinical trial that
norfloxacin, a fluoroquinolone, is an effective alternative to
tetracycline for the treatment of 01 cholera9 and after the
appearance of 0139 cholera, these workers also demon
strated that multiple dose of the same drug is perhaps
superior to doxycycline10. In 1975, scientists from the
NICED, Calcutta, had showed that single dose of
doxycycline (300 mg) is as effective as multiple doses of
tetracycline for the treatment of cholera11. WHO also advo
cates doxycycline for adult cholera. A highly specific mono
clonal antiserum for detection of 0139 strains was devel
oped at NICED which has been supplied to WHO (SEARO),
New Delhi for distribution to various national and interna
tional laboratories.
Studies carried out at NICED showed that cholera
carriere actually act as reservoirs for ElTor cholera vibrios
in the community during the interepidemic period even when
cholera cases are absent12J3. It was further observed that
carriers are responsible for the transmission of V.cholerae
infection to other family members by contaminating unpro
tected stored water and cooked fobd with their infected
fingers14. Transmission studies with V.cholerae 0139 also
showed that at the community level there exists a laige
(14.6%) number of asymptomatic infected persons which
indicates an epidemiological similarity to that of ElTor
cholera15.
In order to find interventions for blocking the transmis
sion of infection at the community level, studies conducted
at NICED showed a lower incidence of cholera carriers
amongst contacts in cholera affected houses provided with
narrow-necked pitchers (sorai)™ and this led to the concept
ofpreventing contamination of stored water by storing water
in narrow necked storage containers which has been widely
accepted. Studies also demonstrated the effectiveness of
simple measures like hand washing with soap and water16,
coupled with health education, in reducing the incidence of
diarrhoeal diseases particularly dysentery cases in Calcutta
slums. The WHO has recommended these strategies for
prevention of diarrhoea.
Development of oral rehydration therapy (ORT) is con
sidered a significant discovery in medicine of this century.
However, such oral rehydration therapy was not much prac
tised particularly for children. A study17 conducted at
NICED on 1974 showed that if repeated small amounts of
oral rehydration solution (ORS) were administered orally at
50
frequent intervals, nearly 91 per cent of diarrhoeal children
below 5 years of age with moderate degree of dehydration
could Be successfully rehydrated by ORS alone. This led to
the widespread acceptance of ORT, particularly for children.
Further studies18 demonstrated that neonates and young
infants could also be safely and effectively rehydrated with
WHO recommended formula of ORS, provided breast feed
ing is continued along with the ORS feeding. It was further
demonstrated19 that ORS of WHO formula was safe and
effective in the treatment of dehydration owing to acute
diarrhoea in malnourished (marasmic) children. Based on
these studies as well the 3-tier implementation strategy
developed by NICED in a community-based study where
mothers were found to play a pivotal role in preventing
dehydration with early institution of “home-available fluid’ ’
therapy, the Government ofIndia in a comprehensive manner
launched the National Diarrhoeal Diseases Control
Programme on a high priority basis with a taiget of reduction
of diarrhoea-related mortality by 50 per cent by 1990.
In view of some limitations of the conventional phage
typing scheme for ElTor vibrios, a much needed new phage
typing scheme for Vibrio cholerae biotype ElTor strains has
been developed at the NICED20. A total of 1000 V.cholerae
01 biotype ElTor strains were phage typed with a set of
10 phages. Over 99 per cent of strains were found to be
typable with this scheme and the strains could be clustered
into 27 types. This new phage typing scheme is expected
to be useful as an epidemiological marker for tracing the
source of infection20.
In a case-control study to understand the risk factors for
development oflife-threatening dehydration, in children upto
the age of 2 years suffering from acute watery diarrhoea, it
was shown21, by univariate analysis, that the presence of
vibrios in the stool, withdrawal of breast feeding during
diarrhoea, not giving fluids, including oral rehydration
solution (ORS) during diarrhoea, frequent purging (>8/day),
vomiting (>2/day) and malnutrition were identified as risk
factors. However, by multivariate analysis, after controlling
the confounders, withdrawal of breast feeding during diar
rhoea, and not giving ORS during diarrhoea were identified
as significant risk factors. It is felt that these findings will
be useful for- both the Global and National Diarrhoeal
Diseases Control Programmes for formulating strategies for
preventing death due to diarrhoeal dehydration.
A pathophysiological study ofcholera undertaken by the
Department of Gastrointestinal Sciences, CMC Hospital,
Vellore, revealed characteristic microvasculature lesions
in rectal and small intestinal mucosa of patients with
™
cholera22,23. The vascular lesion is apparently caused by lipid
A, the toxic moiety ofthe lipopolysaccharide (LPS) antigen.
Biophysical studies, including two-dimensional NMR spec
troscopic analysis of the interaction of LPS with polymyxin
B suggested that pentamidine might be a potential antagonist
of therapeutic value24. The role of short-chain fatty acids in
the absorption ofwater and sodium in the large intestine was
investigated. Since malabsorbed carbohydrates in vitro are
fermtented by the faecal flora of cholera patients, it appears
that cereal-based oral rehydration solutions provide a source
of short chain fatty acids in the large intestine25,26
The molecular mechanism of pathogenesis of cholera,
with emphasis on the bacterial components that mediate
adhesion of the microorganisms to the host intestinal epithe
lium, is an important area researched at the NICED. Since
lectins, a class of proteins capable of binding specifically to
cell surface sugars, are recognized as important candidates
for mediating host-cell interaction, purification and molecu
lar characterization of V. cholerae lectins was undertaken.
A cell-free haemagglutinin (HA) was isolated and purified
to homogeneity and characterized as a disc-like oligomer of
noncovalently associated polypeptides of Mr 62,000 Da27.
The HA recognizes specifically terminal Bl-galactosyl resi
dues of complex glycoproteins and glycolipids. However,
due to intrinsic low affinity for individual galactosyl resi
dues, the HA showed strong avidity only when the sugar
residues are presented in a clustered configuration to enable
multiple interactions with the protein28. A cell-associated
non-fimbrial HA has been isolated by detergent extraction
of intact cells and purified by affinity chromatography on
columns of chitin, a polymer of N-acetyl glucosamine. The
cell-associated HA has been characterized biochemically,
morphologically and antigenically29.
Studies on Shigella
Epidemics of bacillary dysentery due to multiple drug
resistant Shigella dysenteriae type 1 (Shiga bacillus) have
been reported from Central America, Mexico, Bangladesh,
Sri Lanka and Central Africa since 1969. When the disease
spread to West Bengal in 1984 affecting over 3,50,000
people with 3,500. deaths, the NICED Scientists identified
the causative agent as S.dysenteriae type 1, which was
multi-drug resistant30. The local clinicians were not sure
about the drug of choice to treat shigellosis and we recom
mended the use of nalidixic acid which was found to be
highly effective31,32. More recently, norfloxacin, a
fluoroquinolone33,34, which is a nalidixic acid derivative, was
found to be superior to nalidixic acid in the treatment of
Shigellosis caused by nalidixic acid resistant Shigella. A
single dose of ciprofloxacin35 (1 g) was found to be quite
effective for Shigellosis. A team from the NICED.investi
gated a similar epidemic of bacillary dysentery in the
Andaman & Nicobar Islands36 and identified the causative
agent to be multi-drug resistant S.dysenteriae type 1 and
also reported a number of extraintestinal complications
which included among others, leukemoid reaction in about
31 per cent of patients37.
Rapid emergence of multiple drug-resistant strains of
S.dysenteriae has led to a concerted global effort to develop
effective immunoprophylactic measures against the disease.
The humoral immune response of patients convalescing
from shigellosis, against bacterial components like lipopo
lysaccharide (LPS)38 and outer membrane proteins39,40 has
been analyzed by ELISA and immunoblotting. The role of
gut mucosal T and B lymphocytes as well as non-T and nonB cells in cellular immune response to shigellosis was ex
amined in guineapig model41. The subpopulations of lym
phocytes were identified by immunofluorescence using spe
cific monoclonal antibody markers. The efficacy of a 57 kDa
antigenic polypeptide of the outer membrane of
S.dysenteriae 1 in eliciting delayed type hypersensitivity
(DTH) in mice was evaluated42. Porin of S.dysenteriae type
1 was purified to homogeneity and characterized as a
noncovalently associated trimer of polypeptides with Mr 38
kDa43. The pore-forming activity of the protein was demon
strated in prbteoliposomes reconstituted from the protein and
phospholipids. Porin was found, by ELISA and
immundelectron microscopy, to be labeled by antibody,
indicating its surface exposure43. In another study, the
proliferation of murine splenocytes by Shigella LPS was
found to be inhibited by the monosaccharide, L-rhamnose,
indicating specific recognition ofthe oligosaccharide moiety,
presumably by a cell surface receptor44. This is contrary to
the accepted mechanism of endotoxin-induced proliferation
of B cells. Genetic studies on S.dysenteriae 1 showed that
plasmids and specially a 70 kb large plasmid contributed to
the growth characteristics as well as cell wall morphology
of the organisms45.
Studies on Rotavirus
Rotavirus (double stranded segmented RNA virus) is an
important etiological agent of acute diarrhoea in children
below 2 years of age. Rotavirus infection accounts for more
than 22 per cent of hospitalized children with diarrhoea in
Calcutta46,47. However, in Manipur, as high as 90 per cent
of hospitalized babies excrete rotaviruses in the winter
months46,48. A detailed epidemiological study conducted by
the NICED in Manipur over a period of five years revealed
51
a very unusual rotavirus strain associated with an epidemic
and having a characteristic of long electropherotype but
subgroup I in nature suggesting zoonotic transmission of
rotaviruses48. These strains have greater medical signifi
cance than similar strains occasionally isolated earlier, as
such strains had never been implicated in an epidemic before
this study. Further, molecular biological characterization by
cloning and sequencing of the same strain revealed that
genetically this strain was related to the human VP 7G
serotype 2 strain49 but antigenically reacted to bovine VP
7G serotypes 6 and 10 but not with any known major human
VP 7G serotypes 1 to 4. This study suggested that the strain
might have developed by in vivo reassortment between
human and bovine rotaviruses.
In a study carried out at the All India Institute ofMedical
Sciences, New Delhi, the infecting rotavirus strain 116E was
cultivated and the gene 4 and gene 9 characterized. The
strain showed highest homology (92.2%) with a bovine
serotype 10 strain. Neonatal infection induced a significant
rise in IgA titres in saliva and showed a protective efficacy
of 53 per cent in the first 2 years of life50*52.
Studies on Parasitic Diarrhoea
Some parasites, viz. Entamoeba histolytica, Giardia
lamblia and Cryptosporidium are important causes of diar
rhoea. Studies from the NICED identified Cryptosporidium
in 5.6 per cent of faecal samples from children with acute
diarrhoea admitted to local hospitals in Calcutta53,54. Basic
studies revealed the complex nature of E.histolytica and
Giardia antigens55,56, and monoclonal antibodies to 29 kDa
antigen was helpfill in differentiating pathogenic strains
from non-pathogenic strains of E.histolytica51. A 118 kDa
antigen was found tabe highly specific for Indian Giardia
isolates.
Studies on V. parahaemolyticus
Vibrio parahaemolyticus is an important causative
agent of diarrhoea, especially that associated with consump
tion of marine foods. Kanagawa-positive, clinical isolates of
V.parahaemolyticus were found to adhere to rabbit intestinal
epithelial cells with a significantly higher affinity than
Kanagawa-ndgative environmental isolates58. Adherence
was not correlated with cell surface hydrophobicity and
there was no evidence for a role of haemolysin in the
process58. Despite a similar composition of outer membrane
proteins (OMPs) of Kanagawa-positive and Kanagawa
negative strains of V.parahaemolyticus as indicated in SDSPAGE, OMPs of Kanagawa-positive, clinical isolates were
significantly less immunogenic in mice. In comparison to
52
environmental isolates, the OMPs of clinical isolates were
buried to a greater extent59. However, antisera to OMPs of
Kanagawa-positive strains could block adherence of bacte
ria to rabbit intestinal epithelial cells60.
Studies on Typhoid Fever
The widespread and indiscriminate use, often in
subtherapeutic doses of antityphoid drugs (eg,
chloramphenicol for the treatment of any fever or diarrhoea)
in many parts of the developing world probably led to the
emeigence of multidrug resistant Salmonella typhi from
different parts of the country including Calcutta. Scientists
from NICED identified the multidrug resistant S. typhi
strains from typhoid fever patients admitted to the local
hospitals61*63.' In a randomized clinical trial, it was shown
that multidrug resistant typhoid fever responded to
furazolidone in about 86.2 per cent of cases. Furazolidone
is a cheap drug particularly for the people of the developing
countries64.
Studies on Escherichia coli
Different varieties of Escherichia coli are important
causes of acute diarrhoea, particularly in children in the
developing countries. Heat-stable enterotoxin (STa) oiEcoli
triggers intestinal secretion by stimulating the guanylate
cyclase of the enterocytes. Recent studies from the NICED
•revealed that the pathophysiological effect of STa is initiated
by binding with a receptor and stimulation of particulate
guanylate cyclase results from a cascade mechanism asso
ciated with inositol triphosphate Ca++ protein kinase C
signalling system65*68
Investigation of Epidemics
Teams consisting of epidemiologist, microbiologist and
clinicians from the NICED, Calcutta, have investigated a
large number ofepidemics ofacute diarrhoeal diseases in the
whole of the countiy from time to time and identified the
cause and suggested appropriate remedial measures both
as short and long term, thereby extending help to the respec
tive state governments in tackling epidemics of diarrhoeal
diseases.
Very recently, three Institutes, viz., the Institute of
Microbial Technology Chandigarh, the National Institute of
Cholera and Enteric Diseases, Calcutta and the Indian In
stitute of Chemical Biology, Calcutta, have developed a
recombinant cholera vaccine which has been successfully
tested in animal model and awaiting human volunteer trial.
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S.K. Efficacy of norfloxacin for shigellosis : A double-blind
randomised'clinical trial. J Diarr Dis Res 10 : 146, 1992.
Bhattacharya, S.K., Bhattacharya, M.K., DutU, D., Dutta, P.,
Paul, M., Sen, D., Sarkar, S., Saha (Chowdhury) A. and Pal,
S.C. Single-dose ciprofloxacin for shigellosis in adults (Let
ter). J Infect 25 : 117, 1992.
Bhattacharya, S.K., Sinha,A.K., Sen, D., Sengupta, P.G., Lail,
R. and Pal, S.C. Extra-intestinal manifestations of Shigellosis
during an epidemic of bacillary dysentery in Port Blair, Andaman
and Nicobar Islands (India). J Assoc Physicians India, 36:319,
1988.
37.
Sinha, A.K., Bhattacharya, S.K., Sen, D., Sengupta, P.G. and
Pal, S.C. Leukemoid reaction in Shigella dysenteriae type 1
infection. Indian J Med Res 85: 500, 1987.
38.
Sinha, A.K., Bhattacharya, S.K., Saha, M.R., Bhattacharya,
M.K., Sen, D., Ghosh, A.C. and Pal, S.C. Antibody response
to lipopolysaccharide antigen in Shigella dysenteriae type 1
■infection. Indian J Med Res 88: 1, 1988.
39.
40.
54
Roy, S., Das, A.B. and Biswas, T. Antigenicity and antigenic
relatedness of the outer membrane proteins of Shigella spe
cies. Microbios 79: 55, 1994.
Biswas, T., Chakrabarti, M.K., Bhattacharya, M.K., Sen, D.
and Bhattacharya, S.K. Antigenicity and cross-reactivity of
sarkosyl-soluble envelope (inner membrane proteins) of Shi
gella flexneri 2a as recognised by human antibodies. Biomed
Lett 48: 229, 1993.
41.
Sinha, A.K., Chakraborti, M.K. and Chakraborti, S. Gut mucosal
lymphocyte sub-population in the host-defence of Shigella
infected guineapigs. Immunol Lett 32: 65, 1992.
42.
Sinha, A.K., Chakraborti, S. and Chakraborti, M.K. Delayed
hypersensitivity in relation to host cellular immune responses
in Shigella infected mice. Immunol Infect Dis 4: 149, 1994.
43.
Roy, S., Das, A.B., Ghosh, AN. and Biswas, T. Purification,
pore-forming ability and antigenic relatedness of the major
outer membrane protein of Shigella dysenteriae type 1. Infect
Immun 62: 4333, 1994.
44.
Biswas, T., Roy, S. and Banerjee, K.K. L-Rhamnose inhibits
proliferation of murine splenocytes by the lipopolysaccharide
and polysaccharide moiety of Shigella dysenteriae type 1
lipopolysaccharide. Immunology 84: 322, 1995.
45.
Biswas D., Ghosh, A.N. and Kumar, R. Regulation of cell wall
morphology and growth encoded by plasmids in Shigella
dysenteriae 1. World J Microbiol Biotech 11: 280, 1995.
46.
Ghosh, S.K. and Naik, T.N. Evidence for a new rotavirus
subgroup in India. Epidemiol Infect 102: 523, 1989.
47.
Saha, M.R., Sen, D., Datta, R, Dutta, D. and Pal, S.C. Role
of rotavirus as the cause of acute paediatric diarrhoea in
Calcutta. Trans Roy Soc Trop Med Hyg 78: 818, 1984.
48.
Ghosh, S.K. and Naik, T.N. Detection of a l^rge number of
Subgroup I human rotaviruses with a long RNA electropherotypes.
Arch Virol 105: 119, 1989.
49.
Krishnan, T., Burke, B., Shen, S.,Naik, T.N. andDesselberger,
U. Molecular epidemiology of human rotaviruses in Manipur:
Genome analysis of rotaviruses of long electropherotype and
subgroup L Arch Virol 134: 279, 1994.
50.
Bhan, M.K., Lew, J.F., Sazawal, S.r Das, B.K., Gentsch, J.R.
and Glass, R.I. Protection conferred by neonatal rotavirus
infection against subsequent rotavirus diarrhoea. J Infect Dis
166: 282, 1993.
51.
Das, B.K., Gentsch, J.R., Cicirello, H.G., Woods,P.A., Gupta,
A., Ramachandran, M., Kumar, R., Bhan, M.K. and Glass^R.!.
Characterization of rotavirus strains from newborns in New
Delhi, India. J Clin Microbiol 32: 1820, 1994.
52.
Gentsch, J.R., Das, B.K., Jiang, B., Bhan, M.K., and Glass,
R.L Similarity of the VP4 protein of human rotavirus strain
116E to that of the bovine B223 strain. Virology 194: 424,
1993.
53.
Pal, S.K, Bhattacharya, S., Das, P., Choudhuri, R, Dutta, R,
De, S.P., Sen, D., Saha, M.R., Nair, G.B. and Pal, S.C.
Occurrence and significance of Cryptosporidium infection in
Calcutta. Trans Roy Soc Trop Med Hyg 83: 520, 1989.
54.
Das, P., Sengupta, K., Pal, S., Das, D. and Pal, S.C. Biochem
ical and immunological studies on soluble antigens of Enta
moeba histolytica. Parasitol Res 79: 365, 1993.
55.
Das, R, Sengupta, K., Dutta, P., Bhattacharya, M.K., Pal, S.C.
and Bhattacharya, S.K. Significance of Cryptosporidium as an
etiologic agent of acute diarrhoea in Calcutta - A hospital
based study. J Trop Med Hyg 96: 124, 1993.
gh
56.
57.
58.
59.
60.
Chaudhuri, P.P., Pal, S.» Pal, S.C. and Das, P. Studies on
Giardia lamblia trophozoite antigens using Sephacryl S-300
column chromatography, polyacrylamide gel electrophoresis
and enzyme-linked immunosorbent assay. In: Advances in
Giardia Research. Ed. P.M. Wallis and B.R. Hammond,
University of Calgary Press, Calgary, p 191, 1988.
of multi-drug resistant Salmonella typhi in Calcutta. Indian
J Med Res 95: 179, 1992.
63.
Bhattacharya, S.K., Bhattacharya, M.K., Dutta, P., Saha, M.R.,
Dutta, D., Rasaily, R., Saha, A., and Pal, S.C. Multi-resistant
typhoid fever (Letter). Natl Med J India 5 : 41, 1992.
64.
Dutta, P., Rasaily, R., Saha, M.R^ Mitra, U., Manna, B.,
Chakraborty, S. and Mukherjee, A. Randomized clinical trial
of furazolidone for typhoid fever in children. ScandJ Gastroenterol
28: 168, 1993.
65.
Chakrabarti, M.K., Sinha, A.K. and Biswas, T. Adherence of
Vibrioparahaemolyticus to rabbit intestinal epithelial cells in
vitro. FEMS Microbiol Lett 84: 113, 1991.
Banik, N. and Ganguly, U. Stimulation of phosphoinositides
breakdown by heat stable E.coli enterotoxin in rat intestinal
epithelial cells. FEBS Lett 236: 489, 1988.
66.
Biswas, T. and Chakrabarti, M.K. Antigenicity and antigenic
cross-reactivity of outer membrane proteins of Vibrjo
parahaemolyticus. Int J Med Microbiol 281: 475, 1994.
Banik, N.D. and Ganguly, U. Diacylglyceral breakdown in
plasma membrane of rat intestinal epithelial cells. Effect of
E.coli heat stable toxin. FEBS Lett 250: 201, 1989.
67.
Chakrabarti, M.K., Bhattacharya, J., Sinha, A.K. and De, S.P.
Role of outer membrane proteins on the adherence of Vibrio
parahaemolyticus to rabbit intestinal epithelial cells in vitro.
Int J Med Microbiol 282: 436, 1995.
Ghosh Chaudhuri, A , Sen, P.C. and Ganguly, U. Evidence for
protein kinase C stimulation in rat enterocytes pretreated with
heat stable enterotoxin of Escherichia coli. FEMS Microbiol
Lett 110: 185, 1993.
68.
Ghosh Chaudhuri, A. and Ganguly, U. Evidence for stimula
tion of the inositol triphosphate - Ca2+ signalling system in
rat enterocytes by heat stable enterotoxin of Escherichia coli.
Biochem Biophys Acta 1267: 131, 1995.
Sengupta, K., Das, P., Johnson, T.M., Chaudhuri, P.P., Das,
D. and Nair, G.B. Production and characterisation of mono
clonal antibodies against a highly immunogenic fraction of
Entamoeba histolytica (NIH:200) and their application in the
I detection of current amoebic infection. J Euk Microbiol 40:
>722,. 1993.
61.
Bhattacharya, S.K., Bhattacharya, M.K., Rasaily, R. and Deb,
B.C. Treatment of multi-resistant typhoid fever - A perspec
tive. ICMR Bull 23: 67, 1993.
62.
Saha, M.R., Dutta, P., Bhattacharya, S.K., Rasaily, R., Mitra,
U., Dutta, D., Bhattacharya, M.K., and Pal, S.C. Occurrence
The write-up has been contributed by Dr. S.K. Bhattacharya,
Dr. S.P. De, and Dr. Kalyan Banerjee, National Institute of
Cholera and Enteric Diseases, Calcutta.
ABSTRACTS
Some Research Projects Completed Recently
Clinical application of tumour growth fraction associ
ated markers in the radiotherapy of cancer of the uterine
cervix
The study was undertaken on 312 cervical cancer
patients (stage IIB or III) to define the role of tumour growth
fraction and its relationship to the clinical course of the
f disease, as also to know whether cell kinetics had an asso
ciation with response of the tumour to radiotherapy. Tissue
biopsies and smears of exfoliated cells were taken from all
patients before treatment. All pre-treatment biopsies were
analysed for labelling index using bromo deoxy uridine
(BrdUrd) labelling of DNA and Ki 67 antigen expression.
In addition, immunocytochemical localization was carried
out for argyrophillic nucleolar organiser region (AgNOR),
expression of epidermal growth factor (EGF) and its recep
tor (EGF-R), transforming growth factors alpha and beta
(TGFa and TGFp), HPV transforming protein E6,
tumour suppressor protein p53 and the anti-apoptotic prod
uct bcl-2. Ofthe 312 patients, 152 (106 disease free - Group
1, and 46 with residual/recurrent disease - Group 2, after
treatment) were evaluated through their course of radio
therapy and upto a follow up of 12 months and correlation
was made between pre-treatment status of tumour growth
fraction associated markers and clinical outcome after radio
therapy. To study the transition of parameters analyzed
during radiotherapy, cervical smears were obtained before
the therapy, and after 1000, 2000, 3000 and 4000 cGY.
Pre-treatment analysis of AgNORs significantly corre
lated to disease status after treatment. Lower AgNOR counts
were significantly associated with Group 2 patients suggest
ing that increased proliferative activity may be a positive
prognostic indicator. Observations made during various
phases of radiotherapy revealed consistent higher AgNOR
counts in Group 1 patients. However, the other proliferative
associated markers did not show such an association. EGF
and EGF-R showed significant pre-treatment correlations
with the final disease outcome. Both these markers were
expressed more by patients belonging to Group 2, whereas
thp TGFa was expressed more by patients belonging to
Group 1. Group 1 patients mostly showed mild to moderate
expression for TGFp while most Group 2 patients were
negative for this growth factor. This conspicuous difference
55
was also evident during the various phases of radiotherapy.
Pre-treatment expression of bcl-2 was different in the two
groups of patients. Most patients in Group 1 showed mild
expression while in Group 2, the majority showed a mod
erate to intense expression. Similarly, during the various
phases of radiotherapy, Group 2 patients showed consis
tently higher levels of bcl-2.
M. Krishnan Nair
Regional Cancer Centre
Thiruvananthapuram.
Certain basic immune functions in intracranial tumour
patients
In order to elucidate the role of neuralmodulation of
immune response in patients of intracranial tumours,
immune parameters were studied before and 30 days after
treatment in these patients. Normal individuals matched for
age, sex and socio-economic status served as controls.
Patients with benign tumours were treated by surgery
whereas those with malignant tumours were treated by
surgery, radiotherapy and chemotherapy.
It was found that patients with benign and malignant
tumours had higher total leukocyte count before treatment
which decreased significantly after treatment. Lymphocyte
percentage showed a reduction but absolute monocyte count
and monocyte percentage showed an increase in both benign
and malignant tumour patients. There was an elevation in
neutrophil functions as evidenced by increased phagocytic
index, avidity index, NBT (Nitroblue tetrazolium) reduction
and SIC (serum soluble immune complex) index in brain
tumour patients. As compared to controls total T cell and
T4 counts were reduced in both benign and malignant tumour
patients before and after treatment whereas there was an
increase in T8 counts. Serum IgA and IgG levels showed a
decrease and IgM level an increase. Preoperatively altered
immune parameters like lymphocyte percentage, monocyte
percentage, absolute T lympocyte count, T8 count and IgA
levels returned to base-line levels after treatment in benign
tumour patients.
The study confirmed the altered innate, cell mediated
and humoral immunity in brain tumour patients before and
after treatmenet. Most ofthe immune changes observed were
related to cell mediated immunity and were immunosuppres
sive. Though the pituitary region tumours are benign, pa
tients with these tumours showed either marked increase or
decrease in the levels of immune parameters studied than
observed in those with tumours in other areas. The
immunomodulatory effects of malignant tumours were more
marked than those of benign tumours.
A. Namasivayam
Department of Physiology,
Dr. A.L.M. Postgraduate Institute-of
Basic Medical Sciences,
Madras.
85TH ANNIVERSARY CELEBRATIONS OF ICMR
Diarrhoeal diseases was the theme selected for the
dissemination of scientific information to various groups
of individuals in the month of July 1996.
The National Institute of Cholera & Enteric Diseases,
Calcutta has brought out a number of publications on
diarrhoeal diseases which include a booklet on
“Management and Prevention of Diarrhoea : Role of
Mothers” ( English and Hindi); posters on (i) ORS- A
Simple Effective Weapon to fight Diarrhoea (English and
Hindi), (ii) Diarrhoeal Diseases - A Major Global Health
Problem and Management of Diarrhoea Cases (English
and Hindi), (iii) Awareness about Diarrhoea; and a folder
on “Prevention of Diarrhoea”, and a video film on
“Diarrhoeal Diseases”.
Various topics were highlighted by other institutes/
centres during June 1996.The Regional Medical Research
Centre, Bhubaneswar, organized a health awareness
56
campaign at the Headquarters of the Orissa State Branch
of the Indian Red Cross Society (IRCS), Bhubaneswar
on June 6, 1996. The topic of this programme was noncommunicable diseases. Shri I.C.Das, General Secretary
of IRCS, Bhubaneswar, inaugurated this funcion. Dr. K.
Satyanarayana, Director, RMRC, Bhubaneswar, outlined
the role of ICMR and its institutes/centres in tackling
various health problems in the country.
The Vector Control Research Centre (VCRC) Field
Station, Cherthala, Kerala, organized afield demonstration
on the use of medicinal plants at the Ayurveda Medicinal
Plants Herbarium at Thaliaparambu, Poochakal on June
26,1996 by an Ayurveda physician Dr. Sugadhan Vaidhyar
for the IX and X Standard students. Dr. K.H.
Krishnamoorthy, retd. Professor of Botany, JIPMER,
Pondicherry, delivered a guest lecture on “Medicinal
Plants & Ayurveda ” at the VCRC on July 1, 1996.
The Regional Medical Research Centre, Port Blair,
organized various activities over the last few months to
celebrate the 85th anniversary. The Centre organized
various programmes (lectures/seminars) to create aware
ness about leptospirosis among the people and medical
professionals of these Islands. Health camps were
organized among the Shompens, a Mongoloid tribe
of the islands of Great Nicobar, to create health
awareness. Health education material on nutrition, diarr
hoeal diseases and intestinal infections, vector-bome
diseases, leptospirosis, tuberculosis , AIDS, etc. was
displayed in the exhibition organized during the tourism
festival ‘ ‘Islands on the march” in the month of February
this year.
ICMR NEWS
Participation of ICMR Scientists in Scientific Events:
Dr. V.P. Sharma, Director, Malaria Research Centre
(MRC) , Delhi, participated in the III Neem Awareness
Workshop at Mbita, Kenya (June 22-28, 1996).
Medicine, Moscow, Russia in connection with the Modelling
of Transmission Dynamics of Short and Long term
incubating P. vivax Malaria in India (July 6-16,1996).
Dr. G.V. Satyavati, Director-General, ICMR,
participated in the XIX session of the Joint Co-ordinating
Board ofthe Special Programme for Research and Training
in Tropical Diseases at Geneva (June 24-27, 1996).
Dr. D.A. Gadkari, Dy. Director and Officer-in-Charge,
Drs. S. M. Mehendale and R.S. Paranjape, Assistant
Directors and Dr. S.P. Tripathy, Senior Research Officer,
National AIDS Research Institute , Pune, participated in
IX International conference on AIDS at Vancouver, Canada
(July 7-12; 1996).
Dr. K. Vijayaraghavan, Dy. Director, National Institute
of Nutrition (NIN) , Hyderabad, participated in the
Workshop on Consultative Meeting on the Regional
Networking Activities ofthe Regional Training Programme
on Food and Nutrition Planning (RTP-FNP) at Chiang
Mai, Thailand (June 30-July’5,1996).
Dr. I. Khatkhetay, Research. Officer, Institute for
Research in Reproduction, Mumbai; participated in the
International Congress on Clinical Chemistry at London
(July 7-12,1996).
Dr. Padam Singh, Director, Institute for Research in
Medical Statistics, New Delhi, participated in the III
Iranian Statistics Conference at Tehran, Iran (July 1-3,
1996).
Dr. Bela Shah, Dy. Director General, ICMR,
participated in the International conference on “Population
Ageing” and a workshop on “Research Priorities on
Health Ageing” at Brasilia, Brazil (July 1-4, 1996).
Dr. T. Adak, Assistant Director, MRC, Delhi, visited
Martsinox'sky Institute ofMedical Parasitology and Tropical
Dr. G.V. Satyavati, DG, ICMR, participated in the
X meeting of the Joint) Working Group of the Indo-US
Vaccine Action Programme at Washington, D.C. (July
8-15, 1996).
Dr. P. Bhaskaram, Dy. Director, NIN, Hyderabad,
participated in the Indo-UK workshop on Host Parasite
Relationship at Maryland (July 9,1996).
Dr. KamalaKrishnaswamy, Dy. Director (Sr. Grade),
NIN, Hyderabad, participated in the expert panel
meeting on Diet and Cancer Project at London (July
10-12,1996).
ICMR AIDED SYMPOSIA/SEMINARS/WORKSHOPS/COURSES/CONFERENCES
S>ui posium/Semi ni r/Workshop/
Cou rse/Conference
Date & Place
Contact Address
Intemationl Congress on Ageing
August 19-21, 1996;
(at Thiruvananthapuram)
Dr. PJC.B. Nair, Coordinator of the Congress,
Centre for Gerontological Studies, ThiruvananthApuram
Workshop on Design and Evaluation of Targeted Drug
Delivery System
September, 1996;
(at Manipal)
Dr. N. Udupa, Convenor of Workshop, College of
Pharmaceutical Sciences, Manipal.
57
COUNCIL’S TRAINING PROGRAMMES FOR 1996-97
Leprosy
Occupational Health
At the Central Jalma Institute for Leprosy, Agra:
At the National Institute of Occupational Health,
Ahmedabad:
Multidrug Therapy Orientation Course in Leprosy for
Medical Officers (September' 9-20, 1996).
Orientation Course on Occupational Health for Indus
trial Medical Officers (September 17-29, 1996).
Virology
At the National Institute of Virology, Pune:
•
Diploma in Medical Virology (June 1996-May 1997).
Medical Entomology
At the Vector Control Research Centre, Pondicherry:
M.Sc. in Medical Entomology (from August 1996; for
2 years).
Reproductive Biology
At the Institute for Research in Reproduction, Mumbai:
Training Course on Current Trends in Management of
Infertility and Reproductive Disorders (September
2-13, 1996)
Laboratory Animal Technology
At the National Centre for Laboratory Animal Science,
National Institute of Nutrition, Hyderabad:
Training Course for Laboratory Animal Technicians
(June 15-July 31, 1996).
Endocrinology
Training Course for Laboratory Animal Supervisors
(September 2-November 29, 1996.
At the National Institute of Nutrition, Hyderabad:
Annual Training Course On Endocrinological Tech
niques and their Applications (August 1-September
15, 1996).
Haematology
At the Institute of Immunohaematology, Mumbai:
Training Course in Transfusion Medicine for Blood
Bank Medical Officers (August 5-October 4, 1996).
Nutrition
At the National Institute of Nutrition, Hyderabad:
M.Sc. in Applied Nutrition (June 1, 1996-February.
28, 1997).
Training Course in Blood Group Serology and
Blood Bank Methodology for Technicians
(August 5-September 4, 1996).
Annual Training Course in Nutrition (December 1,
1996-February 28, 1997).
* Training Course in Advanced Haematology and
Immunohaematology (September 17-October4,1996).
*
ICMR PUBLICATIONS
Price (Rs.)
Nutritive Value of Indian Foods (1985), by C. Gopalan, B.V. Ramasastri and
S.C. Balasubramaniam, Revised and Updated (1989), by B.S. Narasinga Rao, K.C. Pant
and Y.G. Deosthale (Reprinted 1995)
23.00
Growth & Physical Development of Indian Infants and Children (1972, Reprinted 1989)
10.00
58
Studies on Weaning & Supplementary Foods (1974, Reprinted 1996)
15.00
Studies on Pre-School Children (1974, Reprinted 1984)
6.00
A Manual of Nutrition (Second Edition 1974, Reprinted 1995)
6.00
Low Cost Nutritious Supplements (Second Edition 1975, Reprinted 1994)
4.00
Menus for Low Cost Balanced Diets and School Lunch Programmes Suitable for
North India (Second Edition 1977, Reprinted 1994)
4.50
Some Common Indian Recipes and their Nutritive Value (Fourth Edition 1977,
Reprinted 1995) by Swaran Pasricha & L.M. Rebello
10.00
Nutrition for Mother & Child (Third Edition 1978, Reprinted 1994) by P.S.
Venkatachalam & L.M. Rebello
9.00
Japanese Encephalitis in India (Revised Edition 1980)
5.00
Some Therapeutic Diets (Fourth Edition 1988, Reprinted 1995) by Swaran Pasricha
4.50
Nutrient Requirements & Recommended Dietary Allowances for Indians (1990, Reprinted 1995) 16.00
Count What You Eat (1989, Reprinted 1994) by Swaran Pasricha
9.00
Diet & Diabetes (Second Edition 1993) by T.C. Raghuram, Swaran Pasricha .& R.D. Sharma
18.00
Dietary Tips for the Elderly (1992) by Swaran Pasricha & B.V.S. Thimmayamma
3.50
Diet and Heart Disease (1994, Reprinted 1996) by Ghafoorunissa and Kamala Krishnaswamy
26.00
•Depressive Disease (1986) by A. Venkoba Rao
58.00
♦•Medicinal Plants of India Vol.2 (1987)
136.00
* 10 per cent discount allowed to individuals.
•• 25 per cent discount allowed to individuals.
These publications are available on prepayment of cost by cheque, bank draft or postal order (bank and postal charge
will be extra) in favour of the Director-General, Indian Council of Medical Research, New Delhi. Money orders are no
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.4911, Ansari Nagar, New Delhi-110029 (India).
ERRATA
ICMR Bulletin, Vol.26, No.3&4, March-April, 1996 “Highlights of Tribal Health Research under
Indian Council of Medical Research”
On page 25, column 2, para 3, last sentence — B-thalassaemia mutation should be read as Hb Hofu
[B 126 (HJ Val->Glu]-B°-thalassaemia [Codons 8/9 (+G)]
In para 4, lines 2,5 and 7 — a thalassaemia and a Thai type II should be read as a and B-thalassaemia,
a Thai type II and a thalassaemia, respectively.
ICMR Bulletin, Vol.26, No.6, June 1996 “Control of Iodine Deficiency through Safe Use of Iodized
Salt”
On page 42, column 1, para 2, line 6 “is dependent on the soil content of iodine.” should be read as “is
dependent on the iodine content of soil.”
59
EMERITUS MEDICAL SCIENTISTS
INDIAN COUNCIL OF MEDICAL RESEARCH
Nominations/applications are invited from distinguished retiring scientists engaged in research in the
field of biomedicine for consideration for appointment as Emeritus Medical Scientists under the Council. Full
particulars can be obtained from the office of the Director-General, (Personnel Section), Indian Council of
Medical Research, Post Box No. 4911, New Delhi-110 029. Scientists who have retired or are going to retire
upto 31st March, 1997 may apply for the appointment as Emeritus Medical Scientist under the Council.
Last date for receipt of applications is 30th September, 1996.
INDIAN COUNCIL OF MEDICAL RESEARCH
Grant-in-aid for organising Seminars/Symposia/Workshops
The Council provides partial financial assistance for organising Seminars/Symposia/Workshops.
Applications for grant of financial assistance (complete in all respects in the prescribed proforma), will be
considered only if furnished at least four months before the date of commencement! of the Seminar/
Symposium/Workshop, etc.
EDITORIAL BOARD
Dr. G.V. Satyavati
...
Chairperson
Dr. Badri N. Saxena
...
Member
Dr. N. Medappa
...
Editor
Printed and Published by Shri J.N. Mathur for the Indian Council of Medical Research, New Delhi
at the ICMR Offset Press, New Delhi-110029
R.N. 21813/71
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