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Toxic components of STE induced serious, adverse human oral health outcomes. In the present study, we observed that STE was involved in oral toxicity by reducing the viability of human squamous epithelial cells, SCC-25, along with the simultaneous induction of both apoptosis and autophagic signaling. STE was also found to induce significant amount ROS generation in SCC-25 cells. The dietary flavonoid morin, found abundantly in a variety of herbs, fruits and wine, has been reported to attenuate ROS-induced pathogenesis including autophagy. In this study we designed three different treatment regimes of morin treatment, such as pre, co, and post - treatment of STE challenged SCC-25 cells. In all cases morin provided cytoprotection to STE challenged SCC-25 cells by augmenting STE induced ROS-dependent cytotoxic autophagy. Hence, morin is a potential option for antioxidant therapy in treatment of STE induced toxicity.

Colchicine is a well-known and potent microtubule targeting agent, but the therapeutic value of colchicine against cancer is limited by its toxicity against normal cells. But, there is no report of its cytotoxic potential against lung cancer cell, at clinically permissible or lower concentrations, minimally toxic to non-cancerous cells. Hence, in the present study, we investigated the possible mechanism by which the efficacy of colchicine against lung cancer cells at less toxic dose could be enhanced. Colchicine at clinically admissible concentration of 2.5 nM had no cytotoxic effect and caused no G2/M arrest in A549 cells. However, at this concentration, colchicine strongly hindered the reformation of cold depolymerised interphase and spindle microtubule. Colchicine induced senescence and reactive oxygen species mediated autophagy in A549 cells at this concentration. Autophagy inhibitor 3-methyladenine (3-MA) sensitised the cytotoxicity of colchicine in A549 cells by switching senescence to apoptotic death, and this combination had reduced cytotoxicity to normal lung fibroblast cells (WI38). Together, these findings indicated the possible use of colchicine at clinically relevant dose along with autophagy inhibitor in cancer therapy.

Microtubule-Targeting agents (MTA) are indispensable for cancer therapeutics. We here report thymoquinone (TQ) as a new MTA that already has been appreciated for its anticancer effects. TQ induced G2/M cell cycle arrest in human non-small lung epithelial cells (A549) and majority of arrested cells were in mitosis. TQ depolymerized the microtubule (MT) network and disrupted mitotic spindle organization of A549 cells. MT depolymerization by TQ was followed by apoptosis and subsequent loss in cell viability (IC50 value of ∼10 μM). Interestingly, TQ didn't affect the MT network of normal HUVEC cells at and below the IC50 concentration for A549 cells. TQ also inhibited tubulin polymerization in cell-free system with an IC50 of 27 μM and bound to tubulin heterodimers at a single site with a dissociation constant of 1.19 μM at 25 °C. Binding of TQ to tubulin quenched the tryptophan fluorescence of protein in a time-dependent manner. The TQ-tubulin binding kinetics was biphasic in nature and equilibrated in 30 min. TQ competed with colchicine for tubulin binding with a Ki of 1.9 μM as determined by modified Dixon plot analysis, this suggests TQ may bind tubulin at or near the colchicine binding site and in silico modeling study supported that. Our results establish a novel antimitotic mechanism of TQ by its direct binding to tubulin-MT network in A549 cells.

BACKGROUND High rates of typhoid fever and the emergence of multi-drug resistant strains create a need for prevention efforts including vaccines. Socio-behavioral research can provide important data for participation in future trials and public health vaccination campaigns. DESIGN A 3b phase clinical trial in Kolkata India including pre- and post-vaccination socio-behavioral surveys. RESULTS 47.9% of respondents were male. Ward 29 respondents included 32.4% Hindu and Ward 30 respondents were 99.0% Hindu. Lower rates of participation were found among Muslim respondents and those with post high school education. Lack of information and negative information affected participation. Joint decision-making within households increased participation rates. METHODS seven hundred households were randomly selected 503 respondents (71.85%) completed both the pre- and post-closed-ended surveys. Data analysis included descriptive statistics, Pearson's chi-square tests, independent t-tests, and stepwise logistic regression analysis. Four open-ended questions were included in the survey. These qualitative data were coded and reviewed for common themes and patterns. CONCLUSIONS Individuals' decisions to participate or not participate in a vaccine trial entail a balance between individual beliefs, household dynamics and socio-political influences. Efforts prior to vaccination trials need to develop strategies which address potential underlying mediators for belief systems as well as structural factors which may reinforce individuals' beliefs and perceptions about vaccination trials.

A live oral cholera vaccine developed from a non-toxigenic Vibrio cholerae O1 El Tor strain VA1.3 was tested in a double-blind randomized placebo controlled study for safety and immunogenicity in 304 men aged between 16 and 50 years from Kolkata, India. A dose of 5 x 10(9)CFU (n=186) or a placebo (n=116) containing the diluent buffer was administered. The vaccine did not elicit adverse events except in two vaccine recipients with mild diarrhoea and vomiting. None excreted the vaccine strain. Vibriocidal antibody response developed in 105/186 (57%) and 5/116 (4%) in vaccine and placebo recipients, respectively. In a subgroup, anti-CT antibody rose (or =2-folds) in 23/30 (77%) and 6/19 (32%) in vaccine and placebo recipients, respectively. These studies demonstrate that VA1.3 at a dose of 5 x 10(9) is safe and immunogenic in adults from a cholera endemic region.

The incidence of Vibrio cholerae non-O1, non-O139 strains from hospitalized patients with acute diarrhea constituted 27.4% ( n = 54) of the total 197 V. cholerae strains isolated from patients in Kolkata, India, in 2003. Of 197 strains, 135 were identified as O1 serotype Ogawa and 2 were identified as O139. In the same time period, six O1 background rough strains that possessed all known virulence factors were identified. Serotype analysis of the non-O1, non-O139 strains placed 42 strains into 19 serogroups, while 12 remained O nontypeable (ONT); the existing serotyping scheme involved antisera to 206 serogroups. Detection of a good number of ONT strains suggested that additional serogroups have arisen that need to be added to the current serotyping scheme. The non-O1, non-O139 strains were nontoxigenic except for an O36 strain (SC124), which regulated expression of cholera toxin as O1 classical strains did. Additionally, strain SC124 carried alleles of tcpA and toxT that were different from those of the O1 counterpart, and these were also found in five clonally related strains belonging to different serogroups. Strains carrying tcpA exhibited higher colonization in an animal model compared to those lacking tcpA . PCR-based analyses revealed remarkable variations in the distribution of other virulence factors, including hlyA , rtxA , Vibrio seventh pandemic island I (VSP-I), VSP-II, and type III secretion system (TTSS). Most strains contained hlyA (87%) and rtxA (81.5%) and secreted cytotoxic factors when grown in vitro. Approximately one-third of the strains (31.5%) contained the TTSS gene cluster, and most of these strains were more motile and hemolytic against rabbit erythrocytes. Partial nucleotide sequence analysis of the TTSS-containing strains revealed silent nucleotide mutations within vcsN2 (type III secretion cytoplasmic ATPase), indicating functional conservation of the TTSS apparatus.

India has reasons to be concerned about climate change. Over 650 million people depend on climate-sensitive sectors, such as rain-fed agriculture and forestry, for livelihood and over 973 million people are exposed to vector borne malarial parasites. Projection of climatic factors indicates a wider exposure to malaria for the Indian population in the future. If precautionary measures are not taken and development processes are not managed properly some developmental activities, such as hydro-electric dams and irrigation canal systems, may also exacerbate breeding grounds for malaria. This article integrates climate change and developmental variables in articulating a framework for integrated impact assessment and adaptation responses, with malaria incidence in India as a case study. The climate change variables include temperature, rainfall, humidity, extreme events, and other secondary variables. Development variables are income levels, institutional mechanisms to implement preventive measures, infrastructure development that could promote malarial breeding grounds, and other policies. The case study indicates that sustainable development variables may sometimes reduce the adverse impacts on the system due to climate change alone, while it may sometimes also exacerbate these impacts if the development variables are not managed well and therefore they produce a negative impact on the system. The study concludes that well crafted and well managed developmental policies could result in enhanced resilience of communities and systems, and lower health impacts due to climate change.

SUMMARYIn this study, we have analysed the epidemiological significance of the concurrent infections caused by Vibrio cholerae and intestinal parasites among different age groups of hospitalized diarrhoeal patients in Kolkata. A total of 3556 stool samples collected during 1996–2004 were screened for vibrios and parasites. The seasonality of V. cholerae and parasitic infections were studied in detail. The detection rates for Ascaris lumbricoides and Giardia lamblia infection were more than for the hookworm, Trichuris trichiura and Entamoeba histolytica. V. cholerae O1 was identified as the predominant serogroup among diarrhoeal patients. The highest rates for V. cholerae infection were in the 2–5 years age group and combined infection of V. cholerae and parasites was recorded among children aged between 2 and 10 years.

Picobirnaviruses (PBVs) with bisegmented small RNA genome profile (1.75 and 1.55 kbp for segment 1 and 2, respectively) were detected from 1999 to 2003 in faecal specimens of acute watery diarrhoea cases, largely children ( n = 20) and an adult in Kolkata, India. Varying degrees of dehydration necessitated their visit to hospital for further treatment and management of acute watery diarrhoea. PBV was associated with rotavirus ( n = 3) or astrovirus ( n = 3) and with both in one case. No co-infection with norovirus, sapovirus or adenovirus was detected in the picobirnavirus positive cases. No co-infection with parasites ( Cryptosporidium spp., Giardia spp., Entamoeba spp., helminths) or bacteria ( Vibrio spp., Shigella spp., Escherichia coli ) was detected among the picobirnavirus positive cases. There was a single instance of co-infection with Salmonella spp. ( n = 1). PBVs not associated with serious diarrhoea illness and showing large genome profile (2.3–2.6 and 1.5–1.9 kbp for segment 1 and 2, respectively) have earlier been reported in adult individuals and recently among children from a slum community in Kolkata, India. The short genome profile PBVs associated with acute watery diarrhoea may be another emerging diarrhoeagenic virus in Kolkata, India. Molecular characterization using reported primers PicoB25–PicoB43 for Genogroup I and PicoB23–PicoB24 for Genogroup II in RT-PCR showed the presence of Genogroup I PBVs ( n = 6) and Genogroup II PBVs ( n = 5), while some could not be amplified ( n = 3) with these primers. Sequence analysis of Genogroup I amplicons indicated remarkable sequence heterogeneity. After more than a decade, four PBV positives of Genogroup II were detected during this study. Phylogenetic analysis showed varying degree of genetic diversity amongst PBV strains from Kolkata and other countries.

The study is aimed to determine the seasonal distribution and clinical characteristics of astroviruses associated with acute watery diarrhoea among children in Kolkata and characterize them at the molecular level. Method of study Faecal specimens of acute watery diarrhoea cases (n = 857) and non-diarrhoeic samples (n = 211) from the hospitals and a nearby field community were screened with IDEIA Astrovirus detection kit; astrovirus co-infections with rotavirus and/or picobirnavirus were detected by RNA-PAGE and silver staining. Further RT-PCR was carried out using specific primers, viz. Mon340 (+) and Mon348 (−) targeting a highly conserved domain of ORF1a (289 bp) of human astroviruses. Results Astrovirus infection was detected in 50 cases (50/857); astroviruses were detected mostly in children aged 6–12 months (50%); all non-diarrhoeic samples (n = 211) were negative for astrovirus. In 52% of astrovirus positive cases, the virus was detected as the sole agent; mixed infections were also detected with other diarrhoeic pathogens such as rotavirus (32%), picobirnavirus (2%), rotavirus and picobirnavirus (2%), picobirnavirus and Enterotoxigenic E. coli (ETEC) (2%), rotavirus and ETEC (2%), rotavirus and Enteroaggregative E. coli (EAEC) (2%), Enteropathogenic E. coli (EPEC) (2%), Shigella flexneri type 3a (2%) and Ascaris (2%). RT-PCR and sequencing of amplicons of astroviruses from Kolkata, with specific primers targeted to the conserved domain of ORF1a (289 bp) of the astrovirus genome, showed maximum homology to the astrovirus strain (“5–158”) from Seoul (98%). Results and Conclusions Clinical characteristics of the diarrhoeic children in Kolkata indicated that astrovirus infections were detected throughout the year and were associated with varying degree of dehydration and acute watery diarrhoea. In-depth molecular epidemiological surveillance of astroviruses in Kolkata is essential for better understanding of their overall genetic nature.

Background Rotavirus genotypes, G1–G4 and G9 are associated with childhood diarrhoea throughout the world. In our previous study, we detected G1, G2, G4 and three G12 strains from Kolkata, India. Objectives To study the prevalence of G- and P-genotypes of rotaviruses associated with dehydrating diarrhoea in children admitted to two leading hospitals in eastern India. Study design An active surveillance was conducted for elucidation of rotavirus infection in two leading hospitals in Kolkata, West Bengal and Berhampur (GM), Orissa, India, separated by 603 km from January 2003 to April 2005. The rotaviruses were detected by RNA electrophoresis in polyacrylamide gels. G- and P-typing of the positive samples were accomplished by amplifying VP7 and VP4 genes by RT-PCR and genotyped by seminested multiplex PCR methods. Sequencing, sequence analysis and phylogenetic analysis of VP7 genes of G12 strains were carried out to understand the variations between the strains isolated from different parts of the world. Results The genotypic distribution varied remarkably from our earlier study period (1998–2001) with G1 (53.8%) being the most predominant strain followed by G2 (22.5%), G12 (17.1%), G9 (2.1%) and not a single G3 or G4 isolate was detected separately. 35.2% samples exhibited mixed P-types followed by P[4] (31.7%), P[8] (21.8%) and P[6] (9.8%). The phylogenetic analysis of G12 strains revealed that the G12 strains detected from different parts of the world clustered into three different lineages. Though VP7 sequences of G12 strains isolated from Kolkata and Berhampur are conserved, their P-types were different. Conclusion During this study period we reported emergence of G12 strains as an important pathogen among children in eastern India, thus necessitating its inclusion in future polyvalent vaccine to control rotavirus diarrhoea.

Background Human group B rotavirus was first identified as causative agent of a large outbreak of severe gastroenteritis affecting more than 1 million people, predominantly adults in China in 1982–1983. In spite of serological evidences for the presence of group B rotavirus in many countries of the world, the virus has been detected only from China, India and Bangladesh, where most of the cases were from adults. Objectives To ascertain the role of group B rotavirus as an aetiological agent of diarrhoea among children in Kolkata, India. Study design An active surveillance was conducted for rotavirus infection in children in a leading referral paediatric hospital and a few samples were also collected from adults of another hospital in Kolkata, India over a period of 3 years (2002–2004). After primary screening of rotaviruses by RNA electrophoresis in polyacrylamide gel, 200 of 412 samples negative by PAGE were screened by reverse transcription polymerase chain reaction for group B rotaviruses. The group B rotavirus positives samples were also confirmed by dot-blot hybridization. Result During the study period, we detected 37 (18.5%) sporadic cases of human group B rotavirus infection in children below 3 years of age of which 15 (7.5%) showed mixed infection with group A rotaviruses by RT-PCR. In dot-blot hybridization studies the RNA of all rotavirus positive samples hybridized with the nonisotopic psoralen-biotin labeled total RNA probe generated from a human group B rotavirus CAL-1 strain confirming the samples as group B rotaviruses. Conclusion The shift in age preference of group B rotavirus infection from adult to children and mixed infection of group B and group A rotaviruses reveals the importance of group B rotavirus as an etiological agent of childhood diarrhoea. Therefore, future vaccination strategy should include both group A and B rotaviruses to control rotavirus diarrhoea.

HIV-1 subtype C is the major subtype in India as evidenced from the analysis of specific regions within envelope and gag gene. The matrix protein (p17) of HIV-1 which is involved in several functions like the viral RNA transport, nuclear localization, assembly of pre-integration complex into host nucleus has been used to study the strain diversity among female sexworkers in Calcutta. The gene encoding for the HIV-1 matrix protein, p17 was amplified by nested polymerase chain reaction (PCR) from blood samples of HIV-1 seropositive female sex workers (FSW) in Calcutta, India. Genes of twenty-two samples were sequenced and the phylogenetic analysis with different global strains showed that the majority (seventeen) was clustered with Indian type C. A few samples were found to be close to other C subtypes isolated from South Africa, China and Myanmar. The comparison of Calcutta samples with the samples from other regions of India along with other non-asian subtype C sequences clearly revealed a different cluster of Indian sequences. The two samples, cal 242 and cal 709 was found to be the most divergent type and showed close relatedness with African C subtypes.

This study was undertaken to measure the impact of periodic deworming with albendazole on growth status and incidence of diarrhoea in children aged 2-5 years in an urban setting in India and to assess the feasibility of local health workers implementing the procedures involved. This was a double-blind, placebo-controlled, randomized, community-based intervention trial with 702 children randomly allocated to receive either albendazole or placebo. The two study groups received two doses of albendazole (400 mg) or placebo six months apart. Mean weight increased significantly in the albendazole group compared to the control group at three months, six months and nine months following treatment (P

An investigation was conducted to determine the distribution, virulence gene profile and phenotypes of Shiga toxin-producing Escherichia coli (STEC) strains within a dairy farm in Kolkata, India by characterizing the STEC strains isolated from healthy dairy cow and calf stool samples, raw milk and farm floor swabs from July 2001 to March 2002. Primary screening by multiplex-PCR detected stx1 and stx2, the common virulence genes of STEC, in 18.9% of cow faeces, 32.4% of calf stool samples, 21.6% of farm floor swabs and 4.5% of raw milk samples and viable STEC were recovered from 4.5, 9.9, 8.1 and 1.8% of the corresponding PCR-positive samples. Strains harbouring stx1 (63.3%) and hlyA (53.3%) were frequently detected compared to eae (13.3%). Most of the strains harboured similar sets of reported virulence genes common among isolates from diarrhoea patients. Most of the strains also exhibited multidrug resistance, sorbitol fermentation and produced enterohaemolysin. The randomly amplified polymorphic DNA-PCR (RAPD-PCR) profile of the STEC strains isolated from the farm milieu revealed diverse banding patterns and clonal analysis demonstrated that the strains from different sources were not identical but showed some genetic relatedness. The study demonstrates the potential of dairy farm for housing virulent STEC.

Background :In recent years, hepatitis C virus (HCV) infection is gaining importance in Asian countries. Recent studies conducted in different parts of the world revealed that there is a genotypic correlation of disease severity and treatment outcome. Objectives : A detailed study was carried out to delineate the genotypic distribution of HCV among acute and chronic liver disease patients in Kolkata, a city in eastern India. Study design : Acute and chronic liver disease was diagnosed among patients attending hepatitis clinics in the city. Anti-HCV ELISA was performed on the blood samples of the cases and positive samples were tested for presence of HCV–RNA and genotyping of the samples were carried out by reverse transcription and polymerase chain reaction (RT-PCR) and sequencing. Results : Seroprevalence of HCV infection among acute (11.0%) and chronic (25.3%) hepatitis patients were high and among them 97 (75.8%) and 323 (86.1%) were HCV–RNA positive for acute and chronic hepatitis patients, respectively. Genotyping by PCR showed that the predominant genotype was 3b (42.3%) followed by 3a (28.9%) among acute hepatitis group whereas among chronic hepatitis group, the most prevalent genotypes were 3a (34.7%) and 3b (47.7%). Sequence analysis of the untypeable isolates revealed the presence of a rare subtype 6b. Conclusions : The study revealed very high prevalence of HCV among acute and chronic hepatitis patients with predominance of genotype 3. Subtype 6b was commonly found in Thailand but not in India. The detection of this rare strain of Thai origin reveals the spread of HCV infection from Thailand to other parts of Asia. This observation necessitates further intensive surveillance of HCV infection in India to unravel the distribution of genotypes in the country and to correlate disease severity and treatment outcome to the genotype prevalence. © 2004 Elsevier B.V. All rights reserved.

A comprehensive study was performed to examine incidence, species distribution, drugs sensitivity, virulence genes and molecular fingerprints of Aeromonas species isolated from patients with acute diarrhoea over a period of 2 years in Kolkata, India. Following the Aerokey II scheme, more than 95 % of strains were identified to species level. Seven different species were encountered in this study, with Aeromonas caviae being dominant, followed by Aeromonas hydrophila and Aeromonas veronii biovar sobria. Thirty different serotypes were encountered, with O16, O83 and O85 being dominant, but no serotype was associated specifically with a single species. The majority of Aeromonas strains exhibited multidrug resistance. The alt and act genes, which encode heat-labile cytotonic and cytotoxic enterotoxins, were respectively found in 71.9 and 20.1 % of strains examined. Only 2.4 % of strains carried the heat-stable cytotonic enterotoxin (ast) gene. The hlyA gene was found in 28 % of Aeromonas strains. With few exceptions, genomic diversity of Aeromonas strains belonging to the same serotype was observed by random amplification of polymorphic DNA PCR and ribotyping. Different species of Aeromonas and different clones of Aeromonas species seem to be associated with hospitalized cases of diarrhoea in Kolkata, India.

Since the first report of HIV/AIDS in India in 1986, continuous serosurveillance has been undertaken in all Indian states. Recently, five cases of HIV-2 infection have been detected in Calcutta, situated in the eastern part of India. The full-length envelope gene (2.5 kb) of one of the strains was amplified, cloned, and sequenced. Phylogenetic analysis of the Calcutta HIV-2 envelope revealed a close relatedness to the HIV-2 Rod sequence isolated in offshore Senegal. This strain, however, showed a genetic diversity of 13.5% to other Indian HIV-2 isolates.

Virulence-associated genes and neutral DNA markers of Helicobacter pylori strains from the Santhal and Oroan ethnic minorities of West Bengal, India, were studied. These people have traditionally been quite separate from other Indians and differ culturally, genetically, and linguistically from mainstream Bengalis, whose H. pylori strains have been characterized previously. H. pylori was found in each of 49 study participants, although none had peptic ulcer disease, and was cultured from 31 of them. All strains carried the cag pathogenicity island and potentially toxigenic s1 alleles of vacuolating cytotoxin gene ( vacA ) and were resistant to at least 8 μg of metronidazole per ml. DNA sequence motifs in vacA mid-region m1 alleles, cagA , and an informative insertion or deletion motif next to cagA from these strains were similar to those of strains from ethnic Bengalis. Three mobile elements, IS 605 , IS 607 , and IS Hp608 , were present in 29, 19, and 10%, respectively, of Santhal and Oroan strains, which is similar to their prevalence in Bengali H. pylori. Thus, there is no evidence that the gene pools of H. pylori of these ethnic minorities differ from those of Bengalis from the same region. This relatedness of strains from persons of different ethnicities bears on our understanding of H. pylori transmission between communities and genome evolution.

Long electropherotype with Subgroup I specificity is a common feature of animal rotaviruses. In an epidemic of infantile gastroenteritis in Manipur, India, long but SG I strains predominated in the outbreak in the year 1987-88. One such strain isolated from that region, following the outbreak had G9P [19] specificity. As this is a rare combination, the gene sequences encoding VP4, VP6, VP7, NSP1, NSP2, NSP3, NSP4 and NSP5 of this strain were analyzed. All these genes except VP7 were closely related to porcine rotaviruses (95-99% identity at amino acid level) and clustered with the porcine strains in phylogenetic analysis. In addition, it had subgroup I nature and belonged to NSP4 genotype B which is characteristic of animal rotaviruses. This is the first report of a rotavirus with VP6 and NSP4, two crucial proteins thought to be involved in host range restriction and pathogenicity, were of porcine origin and caused diarrhoea in a human host. Among the genes of this strain sequenced so far, only VP7 had highest identity to human strains at amino acid level. This study suggests reassortment may be occurring between human and other animal strains and some of the reassortant viruses may be virulent to humans.

Diarrhoeal infection caused by Escherichia coli is common in India with occasional outbreaks. However, association of different pathotypes of diarrhoeagenic E. coli (DEC) with the disease and its phenotypic and genotypic characteristics are not fully demonstrated. In this study, E. coli strains from sporadic cases and outbreaks of diarrhoea during 2000-2001 were confirmed as DEC by polymerase chain reaction (PCR) targeting the specific virulence genes. DEC represented by enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC) and enteroaggregative E. coli (EAggEC) were mostly belonged to O serogroups 25, 86a, 114 and 146. The gene astA was frequently detected among ETEC and EAggEC than EPEC. After initial screening of 200 DEC strains with serology and antibiotic susceptibility test, 32 strains representing ETEC, EPEC, and EAggEC isolated from different areas of India were included in the pulsed-field gel electrophoresis (PFGE) analysis. Using the PFGE results, the hierarchical representation of different linkage levels between the DEC strains were determined by unweighed pair-group arithmetic mean (UPGAMA) method. Except for few strains, clonotyping by PFGE revealed no correlation between pathotypes and serogroups as well as the place of isolation of the DEC strains. The prevailing clonal diversity among the different categories of DEC strains suggests that the pathotypes of DEC belonged to diverse clones.

A combined reverse transcriptase-polymerase chain reaction (RT/PCR) was used to produce cDNA of the VP7 gene of rotavirus present in the stool samples. A total of 150 rotavirus positive stool samples were used in this study. Multiplex PCR, using the type specific primers, revealed the presence of G1 (49/150, 32.7%), G2 (27/150, 18%) and G4 (30/150, 20%) genotypes among the samples collected during 1999-2000 from children suffering from acute watery diarrhea. Eighteen samples (12%) were of mixed genotype and the remaining 16 samples (10.6%) could not be typed. Comparative analysis of the full length genes of the representative strains with corresponding genotypes incorporated in the human-rhesus rotavirus tetravalent vaccine (RRV-TV) formulation demonstrates variations of the circulating G1, G2 and G4 strains with the corresponding G genotypes present in the vaccine strain.

We compared putative molecular markers of virulence ( vacA , cagA , and iceA ) of Helicobacter pylori strains isolated from 52 adult duodenal ulcer patients from West Bengal, India, with those of H . pylori strains isolated from 48 adult healthy volunteers from the same region. On the basis of genotyping by PCR, we conclude that the H. pylori strains isolated from the two study groups were indistinguishable and that there are geographic variations in the association of certain putative H. pylori virulence genes with clinical status.

Artemisinin (ART) is a well-known anti-malarial drug, and recently it is shown prospective to selectively kill cancer cells. But low potency makes it inappropriate for use as an anticancer drug. In this study, we modulated the ART-induced autophagy to increase Potency of ART as an anticancer agent. ART reduced the cell viability and colony forming ability of non-small lung carcinoma (A549) cells and it was non-toxic against normal lung (WI38) cells. ART induced autophagy at the early stage of treatment. Pre-treatment with chloroquine (CQ) and followed by ART treatment had synergistic combination index (CI) for cell death. Inhibition of autophagy by CQ pre-treatment led to accumulation of acidic vacuoles (AVOs) which acquainted with unprocessed damage mitochondria that subsequently promoted ROS generation, and resulted releases of Cyt C in cytosol that caused caspase-3 dependent apoptosis cell death in ART-treated A549 cells. Scavenging of ROS by antioxidant N-acetyl-cysteine (NAC) inhibited caspase-3 activity and rescued the cells from apoptosis. Similar effects were observed in other cancer cells SCC25 and MDA-MB-231. The appropriate manipulation of autophagy by using CQ provides a powerful strategy to increase the Potency of selective anticancer property of ART.

This chapter highlights the cholera situation in South Asia and the Bay of Bengal region, the original 'homeland' of cholera. A detailed discussion of cholera outbreaks in individual countries in South-East Asia follows. The countries of the World Health Organization (WHO) SEARO (South-East Asia Region) region are discussed first, followed by discussions about the other countries in South-East Asia that do not fall within the purview of the WHO SEARO classification of the member countries of the region. Therefore, the chapter attempts to provide a comprehensive yet precise outline of the major cholera outbreaks that have occurred in the region over the years.

In Ahmedabad, a major city in the state of Gujarat, India, an outbreak of acute secretory diarrhea caused by Vibrio cholerae O1 Ogawa El Tor, V. cholerae O139, and multiple serotypes of enterotoxigenic Escherichia coli (ETEC) occurred in January 2000. All of the representative V. cholerae O1 and O139 isolates examined harbored the ctxA gene (encoding the A subunit of cholera toxin) and the El Tor variant of the tcpA gene (encoding toxin-coregulated pilus). ETEC isolates of different serotypes were positive for the elt gene, encoding heat-labile enterotoxin. To further understand the molecular characteristics of the pathogens, representative isolates were examined by ribotyping and pulsed-field gel electrophoresis (PFGE). Ribotyping showed that the isolates of V. cholerae O1 Ogawa exhibited a pattern identical to that of the prevailing clone of O1 in areas where cholera is endemic in India, and all of the O139 isolates were identical to the BII clone of V. cholerae O139. PFGE of the representative O1 Ogawa isolates exhibited an identical pattern, comparable to the H pattern of the new clone of O1 reported in Calcutta, India. PFGE analysis of the V. cholerae O139 isolates showed identical patterns, but these differed from the PFGE patterns of O139 isolates reported during 1992 to 1997 in Calcutta. ETEC isolates showed genetic heterogeneity among isolates belonging to the same serotype, although the identical PFGE pattern was also observed among ETEC isolates of different serotypes. Antibiograms of the isolates were unusual, because all of the O139 isolates were resistant to nalidixic acid. Likewise, all of the E. coli isolates showed resistance to ciprofloxacin, norfloxacin, and nalidixic acid. This is a unique outbreak, and we believe that it is the first in which V. cholerae and ETEC were concomitantly involved.

The gene coding for outer capsid protein, VP7 of rotaviruses detected among the children suffering from watery diarrhea in the eastern part of India was studied. The full length gene, coding for VP7 was synthesized from a local strain, WD33, by combined reverse transcriptase–polymerase chain reaction. A total of 42 fecal RNA out of 93 rotavirus-positive samples hybridized with this DNA, indicating the prevalence of this strain in the community. Comparison of the nucleotide and deduced amino acid sequence with the VP7 gene of other serotypes revealed that the local strain,WD33, resembled closely with G1 serotype, the homology being 94 and 97%, respectively. Multiplex PCR using the specific oligonucleotide primers also amplified the G1 specific DNA fragment. The overall change of 16 amino acids was noticed when compared with G1 specific prototype Wa strain of which nine amino acids are within the neutralization domain. However, the phylogenetic tree constructed by comparing the G1 strains from different countries demonstrated that this strain WD33 clustered as a distinct sub-lineage of lineage IV and not with Wa strain, that clustered with lineage III.

In order to assess the extent of genomic diversity among Vibrio cholerae O139 strains, restriction fragment length polymorphisms in two genetic loci, rrn and ctx, were studied. Analysis of 144 strains isolated from different regions of Bangladesh and India between 1992 and 1998 revealed the presence of at least six distinct ribotypes (B-I through B-VI) of which three were new ribotypes, and one of these was represented by a nontoxigenic O139 strain. Strains of ribotypes B-I through B-V shared 11 different CTX genotypes (A through K). Antimicrobial resistance patterns of the strains varied independently of their ribotypes and CTX genotypes. Results of this study suggest that V. cholerae O139 is undergoing rapid genetic changes leading to the origination of new variants, and temporal changes in antimicrobial resistance patterns may be contributing to the selection of different variants.

The scenario of cholera that existed previously changed in 1992 and 1993 with the emergence of toxigenic Vibrio cholerae O139 in India. The genesis of the new serogroup formed the impetus to search for O139 phages in and around the country. A total of five newly isolated phages lytic to V. cholerae O139 strains were used for the development of this phage typing scheme. These phages differed from each other and also differed from the existing O1 phages in their lytic patterns, morphologies, restriction endonuclease digestion profiles, and immunological criteria. With this scheme, 500 V. cholerae O139 strains were evaluated for their phage types, and almost all strains were found to be typeable. The strains clustered into 10 different phage types, of which type 1 (38.2%) was the dominant type, followed by type 2 (22.4%) and type 3 (18%). Additionally, a comparative study of phage types in 1993 and 1994 versus those from 1996 to 1998 for O139 strains showed a higher percentage of phage type 1 (40.5%), followed by type 3 (18.8%) during the period between 1993 and 1994, whereas phage type 2 (32.1%) was the next major type during the period from 1996 to 1998. This scheme comprising five newly isolated phages would be another useful tool in the study of the epidemiology of cholera caused by V. cholerae O139.

Since 1996 infections caused by Vibrio parahaemolyticus have increased globally. Outbreak and/or sporadic cases of diarrhea caused by V. parahaemolyticus have been reported from India Indonesia Japan Taiwan the USA Laos Korea Chile Bangladesh Russia Thailand Spain France and Vietnam. This increase in incidence appears to be related to the emergence of a new clone belonging to the O3:K6 serovar which has pandemic potential. Due to its spread in many countries with identical phenotypic and genotypic features the recently emerged V. parahaemolyticus has now been termed a pandemic strain which can be identified by group-specific GSPCR based on the sequence variation in the toxRS gene. In addition the pandemic strains have a novel open reading frame orf8 which corresponds to a filamentous phage f237. On September 14 2003 an outbreak of diarrhea occurred near Dumdum in Kolkata India. About 200 people had diarrhea after consumption of rice with meat served at the blood donation camp. Twenty-one patients were admitted to the Infectious Diseases Hospital (IDH) Kolkata from this outbreak-affected area. Stool specimens were collected from all the patients and screened for common enteric pathogens within two hours of collection using standard methods. In addition 3--5 Escherichia coli colonies grown in MacConkey agar from each stool specimen were screened for diarrheagenic E. coli (DEC) by PCR.4 In this assay all the tested E. coli isolates were negative for common DEC such as enterotoxigenic E. coli enteropathogenic E. coli and enteroaggregative E. coli. (excerpt)

We report the prevalence of the O139 serogroup in Calcutta, India, after its reemergence in August 1996 and the spread of the reemerged clone to other parts of the country by using previously established molecular markers. Phenotypically, the reemerged Vibrio cholerae O139 displayed a difference compared to those that appeared in late 1992 and 1993 in that the current O139 strains are sensitive to co-trimoxazole. Ribotyping with the enzyme Bgl I produced two rRNA restriction patterns in the O139 strains isolated after August 1996, and these patterns were identical to those exhibited by strains of O139 isolated in 1992. Three clones of V. cholerae O139 are currently prevailing in the country, with strains exhibiting three bands after Hin dIII digestion and hybridization with a ctxA probe being dominant. The reemergence of V. cholerae O139 in Calcutta after a 32-month quiescent period reestablishes the O139 serogroup as an entity which is likely to play a crucial role in the temporal antigenic variations among the serogroups of V. cholerae causing cholera.

There was an inexplicable upsurge in the incidence of non-O1, non-O139 Vibrio cholerae among hospitalized patients admitted to the Infectious Diseases Hospital, Calcutta, India, between February and March 1996. Of the 18 strains of V. cholerae isolated during this period, 15 belonged to the non-O1, non-O139 serogroups (4 belonged to O144, 3 belonged to O11, 1 each belonged to O6, O8, O12, O19, O39, and O58, and 2 strains could not be typed), 2 belonged to the O139 serogroup, and 1 belonged to the O1 serogroup. Cell-free culture supernatants of 13 representative non-O1, non-O139 V. cholerae strains evoked a distinct cytotoxic effect on CHO and HeLa cells, and the strains examined produced the nonmembrane-damaging cytotoxin. By several PCR assays, it was determined that none of the non-O1, non-O139 strains were positive for the ctxA , zot , ace , and tcpA genes and for the genes representing the heat-labile toxin, heat-stable toxin, and verotoxin of Escherichia coli and the various variants of these genes. Studies on the clonality of non-O1, non-O139 V. cholerae strains by restriction fragment length polymorphism (RFLP) analysis of rRNA genes and of other genes ( hlyA , hlyU , hlx , toxR , and attRS1 ) and by pulsed-field gel electrophoresis (PFGE) collectively indicate that the upsurge which occurred in February and March 1996 was caused by strains belonging to different clones. Overall, there was an excellent correlation between the results of ribotyping, RFLP analysis of various genes, and PFGE, with strains belonging to a particular serogroup showing nearly identical restriction patterns and PFGE profiles. It is clear from this study that some serogroups of V. cholerae can cause diarrhea by a mechanism quite different from that of toxigenic V. cholerae O1 and O139, and we have proposed the nomenclature of enteropathogenic V. cholerae to include these serogroups.

After a lapse of 33 months, Vibrio cholerae O139, the new serogroup associated with cholera, has re-emerged in Calcutta, India and has become the dominant serogroup causing cholera from September 1996. In neighbouring Bangladesh, V. cholerae O1 biotype El Tor continues to be the dominant cause of cholera with the O139 serogroup accounting for only a small proportion of cases. Comparison of the phenotypic traits of representative O139 strains from Calcutta and Dhaka isolated between December 1996 and April 1997 showed similar phenotypic traits with the exception that Dhaka O139 strains were susceptible to streptomycin whilst Calcutta O139 strains were resistant. The Dhaka and Calcutta O139 strains displayed identical ribotypes but showed remarkable differences in the structure and organization of the CTX genetic element. In the Dhaka O139 strains, two copies of the CTX element were arranged in tandem and this resembled the pattern displayed by the 1992 epidemic strains of O139. The Calcutta O139 strains, in contrast, carried three copies of the CTX genetic element arranged in tandem with the loss of a conserved BglII restriction site in the RS1 element and the appearance of a new HindIII site in the same region. While there may be other factors, it appears that the reorganization of the CTX genetic element in the Calcutta O139 strains may have contributed to the resurgence of this serogroup in Calcutta.

The Ganges delta has been well known for its high endemicity for cholera for a long time. Several pandemics of cholera originated from this region. A year-long hospital-based surveillance for cholera was undertaken in two major government hospitals in Howrah the twin city to Kolkata (capital of West Bengal India) situated on the banks of the river Ganges. Bi-weekly stool samples were collected from all diarrhoea cases admitted to these two hospitals and were screened for Vibrio cholerae using a standard microbiological technique. The antibiotic susceptibility of the isolates was tested by the Kirby-Bauer disc diffusion technique using commercially available discs. (excerpt)

Thiadiazoles are one of the most widely utilized agents in medicinal chemistry, having a wide range of pharmacologic activity. Microtubules (MTs) have always remained a sought-after target in rapidly proliferating cancer cells. We screened for the growth inhibitory effect of synthetic 5-(3-indolyl)-2-substituted-1,3,4-thiadiazoles on cancer cells and identified NMK-TD-100, as the most potent agent. Cell viability experiments using human cervical carcinoma cell line (HeLa cells) indicated that the IC50 value was 1.42±0.11 µM for NMK-TD-100 for 48 h treatment. In further study, we examined the mode of interaction of NMK-TD-100 with tubulin and unraveled the cellular mechanism responsible for its anti-tumor activity. NMK-TD-100 induced arrest in mitotic phase of cell cycle, caused decline in mitochondrial membrane potential and induced apoptosis in HeLa cells. Immunofluorescence studies using an anti-α-tubulin antibody showed a significant depolymerization of the interphase microtubule network and spindle microtubule in HeLa cells in a concentration-dependent manner. However, the cytotoxicity of NMK-TD-100 towards human peripheral blood mononuclear cells (PBMC) was lower compared to that in cancer cells. Polymerization of tissue purified tubulin into microtubules was inhibited by NMK-TD-100 with an IC50 value of 17.5±0.35 µM. The binding of NMK-TD-100 with tubulin was studied using NMK-TD-100 fluorescence enhancement and intrinsic tryptophan fluorescence of tubulin. The stoichiometry of NMK-TD-100 binding to tubulin is 1:1 (molar ratio) with a dissociation constant of ~1 µM. Fluorescence spectroscopic and molecular modeling data showed that NMK-TD-100 binds to tubulin at a site which is very near to the colchicine binding site. The binding of NMK-TD-100 to tubulin was estimated to be ~10 times faster than that of colchicine. The results indicated that NMK-TD-100 exerted anti-proliferative activity by disrupting microtubule functions through tubulin binding and provided insights into its potential of being a chemotherapeutic agent.

To identify the most common isolates from the corneal ulcers with antimicrobial pattern of bacterial isolates. All patients with suspected corneal ulceration presenting to BP Koirala Institute of Health Sciences from Jan 2004 - Dec 2008 were evaluated. Corneal scraping was performed and processed for direct microscopy, bacteriological and fungal culture. Bacterial isolates were subjected to antimicrobial susceptibility testing. Of 351 specimens examined, growth of etiological agents were obtained in 278 (79.20%). Of these, 113 (40.65%) had pure fungal growth, 108 (38.85%) had pure bacterial growth and 57 (20.50%) had mixed fungal and bacterial infection. The commonest fungal pathogen was Aspergillus spp 50 (33.33%) followed by Fusarium spp 19 (12.66%). Staphylococcal aureus 57 (44.53%) was isolated as commonest bacterial agent. Coagulase Negative Staphylococci 20 (15.6%) was second in the list. Pseudomonas spp 12 (9.40%) was the most common gram negative bacilli isolated. Most of the bacterial isolates were sensitive to commonly used antibiotics. Corneal Ulcer is a common problem in eastern Nepal. Knowledge of both fungal and bacterial agents associated with this condition is of value for the prevention and management of corneal ulcers and their complications.

Cefoperazone is a â-lactam antimicrobial and Sulbactam is an irreversible â-lactamase inhibitor. The objective of this study was to know the susceptibility pattern of gram negative bacilli (GNB) towards cefoperazone-sulbactum. All GNB isolated from different clinical samples during the period of May, 2010 to Aug, 2010 were tested for susceptibility to cefoperazone-sulbactum, meropenem, ceftazidime, cefotaxime, ceftriaxone, chloromphenicol, cotrimoxazole, ampicillin, amikacin, nalidixic acid, ciprofloxacin, carbenicillin and piperacillin using standard Kirby-Bauer disc diffusion antimicrobial susceptibility testing method. The susceptibilities were recorded according to CLSI guidelines. A total of 406 GNB were isolated (urine: 66.7%, pus: 19.2%, and blood: 7.9%). Escherichia coli (54.4%) was most frequently isolated organisms followed by Acinetobacter species (17.7%), Klebsiella pneumoniae (9.1%) and Pseudomonas species (6.1%). Overall, 11.8% of isolates showed resistance to cefoperazone-sulbactam. Frequencies of isolates showing resistance to meropenem and amikacin were 14.7% and 26.25% respectively. Only 3.9% of Escherichia coli isolates showed resistance to cefoperazone-sulbactam. For other organisms, their lowest frequency ranging from 0-20%, exhibited resistance to meropenem. In Pseudomonas spp, in-vitro activity of amikacin was also better as only 11.1% isolates showed resistance to it. This study demonstrated the in-vitro synergistic effect of cefoperazonerazone-sulbactam and meropenem having good activity against GNB compared to the activity of other commonly tested antimicrobials. Cefoperazone-sulbactam can be recommended for the clinical practice against GNB exhibiting resistant to other antimicrobials as it is cheaper alternative to meropenem. Our results also focused on the continuous surveillance of the trends and features of resistance of common antimicrobials.

A modest assessment of noise was made in Calcutta Metro, India's first ever underground tube rail system, to examine if the range of noise levels present could endanger the hearing sensitivity of workers for the Metro. Sound measuring instruments of a sound level meter, an octave band analyzer, and a sound level calibrator were used for measuring the sound pressure levels in platforms of three stations: Esplanade, Kalighat and Tollygunge. The results indicated that the averaged A-weighted SPLs in these stations were in the range of 84-87 dBA. In the coaches of the moving train the Leq values ranged 92-99 dBA and LNP 105-117 dBA, all exceeding the safe limit of day time noise exposure of 55 dBA and 85 dBA of ACGIH. The SPLs at 4,000 Hz in the coaches were also in excess of safe exposure limit of 79 dB. The findings thus posed a potential threat to the workers.

SUMMARYThe distribution and virulence ofVibrio choleraeserogroups other than O1 and O139 in India before, during and after the advent of O139 serogroup was investigated. A total of 68 strains belonging to 31 different ‘O’ serogroups were identified during the study period. With the exception of O53, there was no spatial or temporal clustering of any particular non-O1 non-O139 serogroup at any given place. Two of the 68 strains examined produced cholera toxin (CT) which could only be partially absorbed with anti-CT immunoglobulin G. Tissue culture assay revealed that some of the non-O1 non-O139 strains produced factors which evoked either a cell rounding or cell elongation response depending upon the medium used. This study indicates that serogroups other than O1 and O139 should also be continuously monitored.

In a case-control study to understand the risk factors for development of life-threatening dehydration, a total of 379 children comprising 243 cases (moderate or severe dehydration) and 136 controls (non or mild dehydration) up to 2 years of age suffering from acute watery diarrhoea were studied. By univariate analysis, the presence of vibrios in 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 undernutrition were identified as risk factors. However, by multivariate analysis after controlling for confounders, withdrawal of breast feeding during diarrhoea (odds ratio (OR) = 6.8, p0.00001) and not giving ORS during diarrhoea (OR = 2.1, p0.006) were identified as significant risk factors. The confounding variables which also contributed significantly to increasing the risk were age (or = 12 months; OR = 2.7, p = 0.001), frequent purging (8/day; OR = 4.1, p0.00001), vomiting (2/day; OR = 2.4, p = 0.001) and severe undernutrition (%medianor = 60 weight-for-age of Indian Academy of Paediatrics classification; OR = 3.1, p = 0.001). We feel that these findings will be useful for Global and National Diarrhoeal Diseases Control Programmes for formulating intervention strategies for preventing death due to diarrhoeal dehydration.

Three rare human G12 strains were detected from diarrheic clinical samples of children (

Carcinoma of the gall bladder is the most common biliary tract tumor with higher incidence in females and increasing age. The risk is significantly higher in cholelithiasis. Signet ring cell carcinoma is a rare form of mucinous adenocarcinoma and has a worse prognosis. Early diagnosis is rare. We report a case of signet ring cell carcinoma of the gall bladder in a 70 years old female patient. The gross finding was yellowish white mass measuring 4 x 3.5 cm on cut surface of the gall bladder along with thickened wall. Histopathological examination of the specimen shows the sheets of signet ring cells with lateral spread through the lamina propria and large amount of extracellular mucin. There was metastasis in the retroperitoneum and mesenteric lymph node. The tumor was stage IV (according to TNM staging). Patient died 20th post operative day. Since very few cases have been reported, information regarding the behavior and prognosis of gall bladder carcinoma is limited. However it has been seen that stage III and IV carcinoma usually have worse prognosis.

Inguinal hernias are one of the commonest surgical problems in our set up. It is a protrusion of abdominal cavity contents through the inguinal canal. This is a retrospective study done in the Department of Surgery, Nepal Medical College Teaching Hospital (NMCTH) from January 2009 to December 2010. During this period, 120 patients with painless hernia were enrolled in the study. The age range was from 30 years to 76 years with mean age of presentation at 53 years. Out of 120 patients who underwent mesh repair only 74 patients (61.7%) had visible hernia on standing and 46 patients (38.3%) had palpable impulse on cough. 90 patients (75%) were males and indirect hernia (53.3%) was more common than direct hernia. Out of 120 cases, 30 cases underwent inguinal neurectomy which on follow up had no pain at all. In other 90 cases, neurectomy was not confirmed from the operative note. In these 90 cases, 12 patients had mild pain and 10 patients had moderate to severe pain on 6 months follow up. To conclude, surgical repair is recommended for painless inguinal hernia as most of the patients develop symptoms over time and to avoid undue complications.

Globally, pneumonia is the leading cause of death in young children and burden of disease is disproportionately high in South-East Asia Region of WHO. This review article presents the current status of pneumonia disease burden, risk factors and the ability of health infrastructure to deal with the situation. Literature survey was done for the last 20 years and data from country offices were also collected. The estimated incidence of pneumonia in under five children is 0.36 episodes per child, per year. Risk factors are malnutrition (40% in India), Indoor air pollution, non-breast feeding, chronic obstructive pulmonary disease, etc. Strengthening of health care delivery system for early detection and treatment and as well as minimization of preventable risk factors can avert a large proportion of death due to pneumonia.

Pneumococcal infections are important cause of morbidity and mortality. Knowledge of antimicrobial susceptibility patterns plays important role in the selection of appropriate therapy. Present study was undertaken to analyze the susceptibility patterns of pneumococcal isolates against commonly used antimicrobials with special reference to determination of minimum inhibitory concentration (MIC) of penicillin in a tertiary care hospital in eastern Nepal.Twenty-six strains of S. pneumoniae isolated from various clinical specimens submitted to microbiology laboratory were evaluated. All isolates were tested for antimicrobial susceptibility by disk diffusion method. MIC of penicillin was tested by broth dilution method.Of the total isolates 19 (73%) were from invasive infections. Seven isolates were resistant to cotrimoxazole. No resistance to penicillin was seen in disk diffusion testing. Less susceptibility to penicillin (MIC 0.1-1.0 mg/L) was observed in five (17%) isolates. High level resistance to penicillin was not detected. One isolate was multidrug resistant.S. pneumoniae isolates with intermediate resistance to penicillin prevail in Tertiary Care Hospital in eastern Nepal, causing invasive and noninvasive infections. As intermediate resistance is not detected in routine susceptibility testing, determination of MIC is important. It helps not only in the effective management of life threatening infections but is also essential in continuous monitoring and early detection of resistance. In addition, further study on pneumococcal infections, its antimicrobial resistance profile and correlation with clinical and epidemiological features including serotypes and group prevalence is recommended in future.

a number of studies have shown the association between vitamin A deficiency and the increased risk of diarrhoeal and other childhood morbidities and mortality. However, some studies have raised controversies regarding the reduction of the incidence of diarrhoea after vitamin A supplementation to children. This study was undertaken to evaluate the effectiveness of vitamin A supplementation to young rural children in reducing the incidence of diarrhoea.a double-blind randomized intervention trial was carried out amongst 404 rural children between 6-59 months of age to assess the impact of vitamin A supplementation on morbidity due to diarrhoea. Children aged 6-59 months were enrolled and allocated to receive either 200,000 or 50,000 IU of vitamin A and the same dose was repeated after six months. Morbidity due to diarrhoea was observed by twice-a-week household surveillance, during the subsequent one year of follow up. The incidence of diarrhoea was compared between the two supplemented groups. In addition, the overall incidence of diarrhoea n the two supplemented groups was also compared with the incidence observed during the year preceding supplementation.the incidence of diarrhoea was similar in the two supplemented groups (Incidence Rate Ratio = 1.05. 95% C. I. 0.79-1.40). However, the overall incidence of diarrhoea among all the children in the two supplemented groups (0.56 episodes/child/year) was significantly lower than the incidence before supplementation (1.15 episodes/child/year). The Incidence Rate Ratio was 0.49 with 95% C.I 0.40-0.59.the results of this study indicate that vitamin A supplementation in a dose of 200,000 IU, has no additional advantage over 50,000 IU, at least when the aim is to reduce the incidence of diarrhoea. For control of morbidity due to diarrhoea, vitamin A supplementation in a dose of 50,000 IU every six months appears to be adequate, cost effective and suitable for younger children.

Tuberculosis (TB) is a major cause of morbidity and mortality globally, with most cases occurring in developing countries. The importance of extra pulmonary tuberculosis (EPTB) among all forms of tuberculosis has not yet been ascertained in developing countries.To identify and perform phenotypic characterization of clinically important mycobacterium isolates from extra-pulmonary sites along with drug susceptibility testing.A total of 513 specimens from patients of (EPTB) with varied presentation were studied. Speciation and characterization of isolates were done on the basis of growth and biochemical characteristics. Drug susceptibility testing for mycobacterium isolates was done by proportion method.Fifty four patients clinically suspected to have extra-pulmonary tuberculosis were culture positive for mycobacteria. On characterization 48 of the 54 isolates were identified as M. tuberculosis, 4 as M. bovis and 2 were M. avium/intracelulare. Drug susceptibility testing was done for the both M. tuberculosis as well as M. bovis. In single drug resistance, Streptomycin was highest followed by Isoniazid, Ethambutol and Refampicin. Multi drug resistance (MDR) was found in 6 isolates and it was observed only in group I cases.EPTB accounts for 10-15 percent of all cases of tuberculosis. Mycobacterium was present in 10.5 percent samples. 48 isolates out of 54 samples were found to be M. tuberculosis. The maximum numbers of M. tuberculosis were isolated from lymph node aspiration.

A survey was conducted to study the genetic differentiation among 16 tribal groups of Orissa, Madhya Pradesh, and Maharashtra belonging to different ethnic and linguistic affiliations. Sixteen hundred and fifteen blood samples from both sexes were tested for 5 red cell enzyme systems: ACP, ESD, PGD, GLO, LDH, and Hb pattern. Three hundred and nineteen male individuals were tested for G-6-PD enzyme deficiency. The distribution of the enzyme markers and Hb show a range of variation which are more or less within the Indian range. Cases of homozygous HbSS were detected in all the tribes except 3 tribes in Orissa. Two cases of LDH Cal-1 homozygote were found in two Dravidian language speaking Orissa tribes. The chi 2-values for testing the homogeneity of gene frequencies indicate a non-significant heterogeneity for all alleles in the individual system. Within population diversity seems to be larger than between population diversity. The degree of over all genetic differentiation as measured by GST value is 0.0154 +/- 0.0071.