Diabetes mellitus is an independent risk for gastroesophageal reflux disease among urban African Americans.

An association between gastroesophageal reflux disease (GERD) and diabetes mellitus (DM) has been reported. Studies have not been population-based and have failed to include a representative sample of African American subjects. The aim of the study was to determine if DM is independently associated with GERD among urban African Americans. Single-center, population-based survey utilizing a complex, stratified sampling design. To obtain a simple random sample of the entire African American community, targeted survey zones and hand-delivered invitations were identified. Participating subjects had to be self-described African American, age ≥18. Surveys were completed at a computer terminal assisted by a research coordinator. Four hundred nineteen subjects (weighted sample size of 21 264 [20 888-23 930]). GERD prevalence was 23.7% (95% confidence interval [CI] 23.2-23.9). GERD prevalence was 41.5 % in those with DM versus 20.6 % for those without (P < 0.001). Those with GERD had DM longer but had lower glycohemoglobin levels. The prevalence of ≥2 DM comorbidities was higher in those with GERD (odds ratio [OR] = 2.06; 95% CI 1.71-2.48). In the final model, age >40, DM, increasing body mass index, harmful drinking, and increasing smoking dependence were independently associated with GERD. For DM, there was significant effect modification by gender. In males, the risk was (OR = 4.63; 95% CI 3.96-5.40), while in females, the risk was markedly attenuated (OR = 1.79; 95% CI 1.61-2.00). Among urban African Americans, there is an independent association between DM and GERD that appears to be stronger in men. More information is needed to understand this association.

Population-based assessment of heartburn in urban Black Americans.

Prevalence data for heartburn in the urban Black American community is lacking. In order to estimate prevalence for this community, we analyzed data from an ongoing cohort study in progress at our hospital. Comprehensive interviews allowed for exploration of factors associated with heartburn. Complex, stratified sampling design was the method used. Survey invitations are hand-delivered to random blocks in a single zip code tabulation area. One member per eligible household is invited to complete a computer-based survey. Heartburn was defined as ≥ 3 days/week of symptoms as defined by the Montreal Definition and Classification of gastroesophageal reflux disease. Scaling and weighting factors were utilized to estimate population level prevalence. Multivariate logistic regression was used to identify independent predictor variables for heartburn. Enrolled 379 participants corresponding to a weighted sample size of 22,409 (20,888-23,930) citizens. Demographic characteristics of the sample closely matched those of the entire targeted population. Overall, the weighted prevalence of heartburn ≥ 3 times per week was 17.6% (16.4-18.8%). Variables independently associated with heartburn were body mass index, daily caloric and fat intake, diabetes mellitus (odds ratio = 2.95; 2.59-3.36), cigarette smoking, and alcohol consumption (odds ratio = 2.55; 2.25-2.89). Factors inversely associated included illicit drug use and increased physical activity. Waist : hip ratio showed no relationship. The prevalence of heartburn ≥ 3 times per week is high in the Black American community. Adverse lifestyle behaviors showed particularly important associations. Our study needs to be replicated in other communities with similar demographics.

Role of probiotic in preventing acute diarrhoea in children: a community-based, randomized, double-blind placebo-controlled field trial in an urban slum.

Acute diarrhoea remains a major public health challenge in developing countries. We examined the role of a probiotic in the prevention of acute diarrhoea to discover if there was an effect directed towards a specific aetiology. A double-blind, randomized, controlled field trial involving 3758 children aged 1-5 years was conducted in an urban slum community in Kolkata, India. Participants were given either a probiotic drink containing Lactobacillus casei strain Shirota or a nutrient drink daily for 12 weeks. They were followed up for another 12 weeks. The primary outcome of this study was the occurrence of first episodes of diarrhoea. We assessed this during 12 weeks of intake of study agent and also for 12 weeks of follow-up. There were 608 subjects with diarrhoea in the probiotic group and 674 subjects in the nutrient group during the study period of 24 weeks. The level of protective efficacy for the probiotic was 14% (95% confidence interval 4-23, P<0·01 in adjusted model). The reduced occurrence of acute diarrhoea in the probiotic group compared to nutrient group was not associated with any specific aetiology. No adverse event was observed in children of either probiotic or nutrient groups. The study suggests that daily intake of a probiotic drink can play a role in prevention of acute diarrhoea in young children in a community setting of a developing country.

Toxigenic Vibrio cholerae from environmental sources associated with the cholera outbreak after 'AILA' cyclone in West Bengal, India.

AIMS: West Bengal experienced a devastating storm named 'AILA' in its coastal and southern districts. We attempted to understand the transmission dynamics emphasizing on potable water to detect the presence of toxigenic strains of Vibrio cholerae, followed by the natural devastation. METHODS AND RESULTS: A total of 33 water samples (from tap, tube well and ponds) were analysed. From them, 11 (33.3%) samples were found to be contaminated with V. cholerae, among which 5 (45%) isolates were V. cholerae O1 biotype Ogawa. Antibiogram profile reveals that most of the V. cholerae O1 isolates were highly sensitive against fluroquinolone group of antibiotics, but less sensitive against cefuroxime (50%), cefotaxime (40.9%), ceftriaxone (38.63%), trimethoprim (37.3%), streptomycin (29.2%) and furazolidon (4.54%). Three (36%) V. cholerae isolates were found to be ctxB positive (2 ctxB classical). CONCLUSIONS: Potable water plays a crucial role in cholera transmission. Natural disasters like the reported one aided with feacal-oral contamination enhance the possibilities of drinking water contamination. SIGNIFICANCE AND IMPACT OF THE STUDY: The application of the modified technique, making use of the enrichment subsequently followed by culture and PCR, will help us to detect the presence of toxigenic V. cholerae contamination in different aquatic environment. Moreover, natural extremes have a direct role in increase of salinity level, followed by higher predominance of V. cholerae along with their toxicity development in terms of genetic modification.

Prediction and Visualisation of Bony Impingement for Subject Specific Total Hip Arthroplasty.

Bony impingement (BI) may contribute to restricted hip joint motion, and recurrent dislocation after total hip arthroplasty (THA), and therefore, should be avoided where possible. However, BI risk assessment is generally performed intra-operatively by surgeons, which is partially subjective and qualitative. Therefore, the aim of the study was to develop a method for identifying subject-specific BI, and subsequently, visualising BI area on native bone anatomy to highlight the amount of bone should be resected. Activity definitions and subject-specific bone geometries, constructed from CT scans, with planned implants were used as inputs for the method. For each activity, a conical clearance angle (CCA) was checked between femur and pelvis through simulation. Simultaneously, BI boundary and area were automatically calculated using ray intersection and region growing algorithm respectively. The potential use of the developed method was explained through a case study using an anonymised pre-THA patient data. Two pure (flexion, and extension) and two combined hip joint motions (internal and external rotation at flexion and extension respectively) were considered as activities. BI area were represented in two ways: (a) CCA specific where BI area for each activity with different CCAs was highlighted, (b) activity specific where BI area for all activities with a particular CCA was presented. Result showed that BI area between the femoral and pelvic parts was clearly identified so that the pre-operative surgical plan could be adjusted to minimise impingement. Therefore, this method could potentially be used to examine the effect of different preoperative plans and hip motion on BI, and to guide bony resection during THA surgery.

Epidemiological, microbiological & electron microscopic study of a cholera outbreak in a Kolkata slum community.

BACKGROUND & OBJECTIVE: Epidemics of cholera caused by Vibrio cholerae O1 or O139 have been reported from different parts of India. Factors such as unsafe water supply, poor environmental sanitation, indiscriminate defaecation and lack of personal hygiene are mainly responsible for continued transmission of this disease. We report here epidemiological and microbiological findings of a localized outbreak of cholera, which occurred during March and April 2004 in the eastern part of Kolkata city. METHODS: The affected slum area has a population of 4409, predominantly muslims. Patients suffering from acute watery diarrhoea attended the health outposts organized by National Institute of Cholera and Enteric Diseases, Kolkata and International Vaccine Institute, South Korea as part of a routine surveillance programme at the locality as well as the emergency medical camp organized by the Kolkata Municipal Corporation. Stool and water samples were collected and tested for diarrhoeagenic pathogens in the laboratory. Bacteriophages specific for V. cholerae were isolates and studied electron microscopically for morphology. RESULTS: A total of 89 diarrhoea cases were reported giving an attack rate of 2 per cent. V. cholerae O1 biotype ElTor, serotype Ogawa was isolated as a sole pathogen from 15 (15.8%) of 89 stool samples screened. Water samples (2 from tube wells, 3 from municipal taps and 1 from well) showed presence of coliform bacilli with high MPN (Most Probable Number) count. Bacteriophages specific to V. cholerae were isolated from 2 of 6 water samples examined. A leakage was detected in the main pipeline supplying drinking water to that area. INTERPRETATION & CONCLUSION: The outbreak was caused by V. cholerae O1 (Ogawa) biotype ElTor. The presence of phages in the water samples was an additional indicator for V. cholerae contamination in this community. Occurrences of such outbreaks support vaccination against cholera as an alternative strategy.

Mapping of risk prone areas of kala-azar (Visceral leishmaniasis) in parts of Bihar State, India: an RS and GIS approach.

BACKGROUND & OBJECTIVES: The kala-azar fever (Visceral leishmaniasis) is continuing unabated in India for over a century, now being largely confined to the eastern part of India mainly in Bihar state and to some extent in its bordering states like West Bengal and Uttar Pradesh. Two study sites namely Patepur block in Vaishali district with high endemicity in northern part and Lohardagga block in Lohardagga district with absolute non-endemicity in southern part of Bihar were selected for the study with the following objectives : (i) to study the macro-ecosystem in relation to distribution of vector -Phlebotomus argentipes; (ii) to identify/map the risk prone areas or villages in a block for quick remedial measures; and (iii) to make use of satellite remote sensing and GIS to demonstrate the utility for rapid assessment of landuse/landcover and their relation with the incidence of kalaazar leading to the mapping of risk prone areas. METHODS: Indian Remote Sensing (IRS)-1D LISS III satellite data for the periods of March and November 2000 were analysed in Silicon graphic image processing system using ERDAS software. False color composites (FCC) were generated and landuse/landcover was assessed using Maximum likelihood supervised classification techniques based on ground truth training sets. During the study the GIS functions are used to quantify the remotely sensed landscape proportions of 5 km2 buffer surrounding each known group of villages of high occurrence of sandflies in endemic and nonendemic study sites. Instead of traditional ground based survey methods to vector surveillance, the present study used a combination of remote sensing (RS) and geographical information system (GIS) approach to develop landscape predictors of sandfly abundance-an indicator of human vector contact and as a measure of risk prone areas. RESULTS: Statistical analysis using the remotely sensed landscape variables showed that rural villages surrounded by higher proportion of transitional swamps with soft stemmed edible plants and banana, sugarcane plantations had higher sandfly abundance and would, therefore, be at higher risk prone areas for man-vector contact. INTERPRETATION & CONCLUSION: The present study clearly brought out the usefulness of satellite remote sensing technology in generating the crucial information on spatial distribution of landuse/landcover classes with special emphasis on indicator landcover classes thereby helping in prioritising the area to identify risk prone areas of kala-azar through GIS application tools.

Kinetics of conformational changes associated with potassium binding to and release from Na+/K(+)-ATPase.

The Na+/K(+)-ATPase functions in cells to couple energy from the hydrolysis of ATP to the transport Na+ out and K+ in. The fluorescent probe IAF (iodoacetamidofluorescein) covalently binds to this enzyme, reporting conformational changes without inhibiting enzyme activity. This paper describes experiments using dog kidney enzyme labeled with IAF to examine kinetics of conformational changes resulting from added Na+ and K+, measured in terms of steady-state and stopped-flow fluorescence changes. Kinetics of these fluorescence changes were examined as a function of temperature from two initial conditions: (a) enzyme in the high-fluorescence form (E(high)) was rapidly mixed with varying [K+]; and (b) enzyme in the low-fluorescence form (E(low)) was rapidly mixed with varying [ATP]. These experiments showed: (1) The rate constant for the fluorescence change from E(high) to E(low) was much larger than that for the opposite transition, E(low) to E(high); (2) the apparent free energy of activation (Ea(app)) for the two transitions were different (as estimated from Arrhenius plots); (3) under steady-state conditions, IAF fluorescence did not change when ATP was added to E(low)(K+) in the absence of Na+; (4) the apparent free energy of activation was independent of [K+] for the E(high) to E(low) transition (at 16.4 kcal/mol) but increased with [ATP] for the E(low) to E(high) transition; (5) Ea(app) for the E(low) to E(high) transition with 1 mM ATP was approximately the same as that in the absence of ATP (34 kcal/mol). These results can be interpreted as: (i) in the transition from E(low) to E(high), IAF reported a conformational change that occurred after K+ release to the intracellular side and which is involved in Na+ binding; (ii) Ea(app) increased with [ATP], while increasing the entropy of the transition state. Thus, ATP appeared to destabilize the enzyme during the transition from E(low) to E(high).

Role of biomodulators and involvement of protein tyrosine kinase on stem cell migration in normal and leukaemic mice.

Tyrosine kinase has an important role with regard to self-renewal and as a comitogen in the movement of stem cells out of the haemopoietic stem cell pool into the progeny pool. The present investigation has an objective to evaluate the protein tyrosine kinase (PTK) activity of bone marrow derived pluripotent cells before and after application of biological response modifiers (BRMs) in normal and leukaemic mice. The PTK activity of the cytosolic fraction of bone marrow cells has been determined by the assay kit based on per-oxidase labeled substrate analog and biotin-streptavidin expression. A consequent cell population kinetic study has also been conducted. Results showed a higher activity in the cells of leukaemic mice, which under the influence of interleukin-2 (IL-2) and the non-specific BRM sheep erythrocytes (SRBC) undergo further activation. Interferon-gamma (IFN-gamma) when administered alone showed a suppressive effect and the combination of the three manifested a resultant suppression. Corresponding migration (cell population kinetics) of the bone marrow cells (BMC) also correlated well with the PTK activity of the cells concerned. The observations indicated that the pluripotent BMCs are under regulated control of the PTK activity, which can be manipulated by selective BRMs. The data also suggested the therapeutic benefit of IFN-gamma along with chemotherapeutics against leukaemia and that of IL-2 and SRBC during bone marrow failure.

Facilitation of functional compartmentalization of bone marrow cells in leukemic mice by biological response modifiers: an immunotherapeutic approach.

Biological Response Modifiers (BRMs) including interleukin-2 (IL-2), interferon-gamma (IFN-gamma) and sheep erythrocytes (SRBC) protected N,N'-ethylnitrosourea (ENU) induced leukaemic mice. Two cell types from the bone marrow were isolated in density specific gradient representing two distinct compartments, the low density cells being more CD34 positive than the high density group. Investigations with the functional efficacy of such compartments revealed significant improvement of cytotoxic efficacy and phagocytic burst at the high density compartment (HDC) level. The high density compartment was found to be more responsive towards the BRMs compared to the cells of the low density compartment (LDC). It was suggested that use of BRMs in vivo can stimulate a potent functional progenitor compartmentalization in normal as well as leukaemic mice. These observations are expected to help a logistic approach towards combined BRM therapy at the clinical level.

Genus-specific leptospiral antigen and its possible use in laboratory diagnosis.

Sheep red cells, sensitized with a genus-specific leptospiral substance extracted from a water leptospira, were preserved by freeze drying. The serological reactivity of the preparation was tested against a total of 41 sera from proven cases of human leptospirosis and 27 from proven cases of animal leptospirosis. The sensitized, freeze-dried red cells appeared to be a sensitive reagent for the detection of leptospiral antibodies by haemagglutination. The use of this reagent as a screening test for leptospiral diagnosis is discussed. In the reported haemagglutination test on human and rabbit antisera the findings suggested that only IgM (19S) antibodies took part in the reaction.

Transient kinetics of substrate binding to Na+/K(+)-ATPase measured by fluorescence quenching.

This paper examines the transient kinetics of substrate binding to the Na+/K(+)-ATPase labelled with iodoacetamidofluorescein (IAF) using fluorescence quenching by trinitrophenyl-ATP (TNP-ATP). Earlier work (E.H. Hellen, P.R. Pratap, 1996, Fluorescence quenching of IAF-Na+/K(+)-ATPase via energy transfer to TNP-labelled nucleotide, Proceedings of the VIIIth International Conference on the Na+/K(+)-ATPase, in press) has shown that TNP-nucleotide binds to specific sites (from which unlabelled nucleotide can displace it) and nonspecific sites (from which unlabelled nucleotide cannot displace it). Under stopped-flow conditions, quenching of IAF-enzyme fluorescence was well described by a stretched exponential (F(t) = F infinity + delta F exp[-Bt alpha]). Physically, this function may be interpreted in terms of its inverse Laplace transform phi (k), which describes a distribution of rate-constants; alpha reflects the width of this distribution. As TNP-ATP concentration increased, alpha decreased, reflecting TNP-ATP binding to sites with higher energy barriers. alpha decreased by about the same amount with increasing [TNP-ATP] in the presence of saturating ATP, indicating that the distribution of rate-constants is largely associated with the nonspecific binding sites. However, alpha was significantly less than 1 for ATP-induced fluorescence recovery in the presence of TNP-ATP, indicating that rate-constants associated with specific binding site are also distributed. The distribution of rate-constants for binding to the specific site indicates a distribution in the energy of the transition state for substrate binding. These results suggest that the specific binding site (in either the empty or the full state) may exist in a series of conformations separated by small energy barriers. However, the energy barriers for binding associated with these conformations are significantly distributed.

Endometroid adenocarcinoma of the cervix in a 9-year-old girl.

We present the first reported case of endometroid adenocarcinoma of the uterine cervix in a young girl. The differential diagnosis of a vaginal mass in this age group is usually rhabdomyosarcoma, although other, rarer tumours also occur.

A prospective study of phage types & biotypes of Salmonella enterica serotype Typhi isolated from hospitalized children in Kolkata, India.

BACKGROUND & OBJECTIVES: Kolkata and its suburbs in eastern India faced an epidemic of typhoid fever in 1990. A prospective, hospital and laboratory based study over a period of 12 yr (1990-2001), on the phage typing and biotyping pattern of Salmonella enterica serotype Typhi was carried out, to see if there has been a change. METHODS: A total of 338 S. enterica serotype Typhi isolates from 1491 blood samples were phage typed and biotyped. The mean age of isolation was calculated. RESULTS: The age distribution of subjects (neonates to 12 yr) has been analysed. Of the 338 (22.7%) isolates obtained, eight different S. enterica serotype Typhi phage types were detected. Biotype I (95.8%) was more prevalent as compared to biotype II (4.1%). Phage type E1 was the commonest phage type in Kolkata and its suburbs. INTERPRETATION & CONCLUSION: The mean age at isolation was found to be 6.7 +/- 3.3 yr. Biotype I was predominant and it was of interest that all strains of phage type E1 belonged to biotype I.

Field trial of an ecological approach for the control of Phlebotomus argentipes using mud & lime plaster.

A pilot study for the control of Ph. argentipes, a known vector of kala-azar in India, was carried out using an ecological approach. Of the 15 houses selected for the study 10, including the cattle sheds and latrines, were plastered with a mixture of mud and lime, up to a height of 1.22 m taking care to seal all cracks and crevices. The remaining five houses were left unplastered and were considered as control areas. The pre-treatment and post-treatment resting densities of the sandfly were monitored both in treated and untreated houses. A sudden drop in the sandfly density was noticed in the treated houses, whereas there was no significant reduction in the check houses, suggesting an effective control.

The influence of maternal antibody and age of calves on effective vaccination against Leptospira interrogans serovar hardjo.

Twelve seronegative cows were vaccinated with an experimental bivalent Leptospira interrogans serovars hardjo and pomona vaccine late in their first pregnancy. Calves born of these dams were divided into 4 equal groups that received this vaccine at 4, 6, 10 and 18 weeks of age, respectively. Before vaccination the group geometric mean titres of maternally-derived circulating antibodies ranged from 2 to 25 for the microscopic agglutination (MA) test and 3 to 35 for enzyme-linked immunosorbent assay (ELISA) using a serovar hardjo outer envelope antigen. Post-vaccination peak titres were 645 to 1612 for MA and 562 to 1037 for ELISA, respectively. Calves vaccinated at the youngest age, had the highest pre-vaccination circulating maternal antibody titres, but showed the smallest rise in post-vaccination antibody titres. Circulating maternal antibody was detected in calves up to 13 weeks of age. All immunised calves were protected against a virulent challenge with serovar hardjo type Hardjobovis, regardless of their age or maternally-derived antibody titres. These findings indicate that calves as young as 4 weeks old, vaccinated in the presence of maternally-derived antibody, can be fully protected against homologous virulent challenge.

An entomological field evaluation of larval biology of sandfly in Kala-azar endemic focus of Bihar--exploration of larval control tool.

Knowing the exact breeding places inside the habitat is very important to plan the larval control strategy. Information regarding larval biology in relation to different seasons will be more useful to organize insecticide spray schedule at a particular month of maximum immature density to bring down the adult sandfly density. In the present study, maximum number of soil samples were found positive in the month of January and minimum in the month of September. Maximum positive soil samples were collected from cattle sheds, minimum in mixed dwellings and in case of human dwellings all soil samples were negative. Comparison of two methods for the isolation of immature stages showed that direct microscopic observation is superior to sugar flotation technique.

Vectorial efficacy of Phlebotomus argentipes in Kala-azar endemic foci of Bihar (India) under natural and artificial conditions.

Ability of Phlebotomus argentipes to acquire Leishmania donovani the causative agent of Indian Kala-azar was evaluated in the laboratory. Flies were fed artificially on infected blood suspensions, using a chick-skin-membrane feeding apparatus, and naturally on Leishmania donovani infected mice. In addition flies collected from different endemic areas were dissected and examined for natural infection. Flies fed on infected mice showed significantly higher feeding rate (14.4%, p < 0.01) compared to that of other experiments (9%, 8.75%) but the percentage of infection was very low (2.43%). No Chi-square comparison was made between infection rate and feeding rate because of low value in infection rate (less than 5). Flies dissected for natural infection showed only 0.1% infection. Not much difference was observed in the intensity of Leishmania donovani infection in the mid gut of sandflies examined from any of these experiments. These observations have confirmed that Phlebotomus argentipes has ability to acquire infection and it provides the final piece of evidence that Phlebotomus argentipes is the vector of Leishmania donovani in Bihar State.