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Background: Environmental enteric dysfunction (EED) is an asymptomatic intestinal disorder associated with growth impairment, delayed neurocognitive development, and impaired oral vaccine responses., Objectives: We set out to develop and validate a histopathologic scoring system on duodenal biopsies from a cohort study of children with growth failure in Bangladesh, Pakistan, and Zambia ("EED") with reference to biopsies from United States children with no clinically reported histologic pathology (referred to hereafter as "normal") or celiac disease., Methods: Five gastrointestinal pathologists evaluated 745 hematoxylin and eosin slide images from 291 children with EED (mean age: 1.6 y) and 66 United States children (mean age: 6.8 y). Histomorphologic features (i.e., villus/crypt architecture, goblet cells, epithelial and lamina propria acute/chronic inflammation, Brunner's glands, Paneth cells, epithelial detachment, enterocyte injury, and foveolar metaplasia) were used to score each histopathologic slide. Generalized estimating equations were used to determine differences between EED, normal, and celiac disease, and receiver operating characteristic curves were used to assess predictive value., Results: Biopsies from the duodenal bulb showed higher intramucosal Brunner's gland scores and lower intraepithelial lymphocyte scores than from the second or third parts of the duodenum (D2/3), so only D2/3 were included in the final analysis. Although 7 parameters differed significantly between EED and normal biopsies in regression models, only 5 (blunted villus architecture, increased intraepithelial lymphocytosis, goblet cell depletion, Paneth cell depletion, and reduced intramucosal Brunner's glands) were required to create a total score percentage (TSP-5) that correctly identified EED against normal biopsies (AUC: 0.992; 95% CI: 0.983, 0.998). Geographic comparisons showed more severe goblet cell depletion in Bangladesh and more marked intraepithelial lymphocytosis in Pakistan., Conclusions: This scoring system involving 5 histologic parameters demonstrates very high discrimination between EED and normal biopsies, indicating that this scoring system can be applied with confidence to studies of intestinal biopsies in EED., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Background: Environmental enteric dysfunction (EED), a chronic inflammatory condition of the small intestine, is an important driver of childhood malnutrition globally. Quantifying intestinal morphology in EED allows for exploration of its association with functional and disease outcomes., Objectives: We sought to define morphometric characteristics of childhood EED and determine whether morphology features were associated with disease pathophysiology., Methods: Morphometric measurements and histology were assessed on duodenal biopsy slides for this cross-sectional study from children with EED in Bangladesh, Pakistan, and Zambia (n = 69), and those with no pathologic abnormality (NPA; n = 8) or celiac disease (n = 18) in North America. Immunohistochemistry was also conducted on 46, 8, and 18 biopsy slides, respectively. Linear mixed-effects regression models were used to reveal morphometric differences between EED compared with NPA or celiac disease and identify associations between morphometry and histology or immunohistochemistry among children with EED., Results: In duodenal biopsies, median EED villus height (248 μm), crypt depth (299 μm), and villus:crypt (V:C) ratio (0.9) values ranged between those of NPA (396 μm villus height; 246 μm crypt depth; 1.6 V:C ratio) and celiac disease (208 μm villus height; 365 μm crypt depth; 0.5 V:C ratio). Among EED biopsy slides, morphometric assessments were not associated with histologic parameters or immunohistochemical markers, other than pathologist-determined subjective semiquantitative villus architecture., Conclusions: Morphometric analysis of duodenal biopsy slides across geographies identified morphologic features of EED, specifically short villi, elongated crypts, and a smaller V:C ratio relative to NPA slides, although not as severe as in celiac slides. Morphometry did not explain other EED features, suggesting that EED histopathologic processes may be operating independently of morphology. Although acknowledging the challenges with obtaining relevant tissue, these data form the basis for further assessments of the role of morphometry in EED., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

De Quervain's disease (DQD) is commonly reported in mothers during pregnancy up to delayed postpartum period. A cross-sectional study was conducted to assess infant caregivers who visited the paediatric outpatient department or vaccination centre in two hospitals of Lahore, during the months of May and June, 2021. A total of 190 subjects were interviewed directly and assessed by applying Finkelstein's test on both hands. Data was collected using Numeric Pain Rating Scale (NPRS) and Patient Rated Wrist Evaluation (PRWE) from positive subjects. They were asked to report their pain and difficulty level of the affected hand with worsened symptoms. The results exhibited 26.8% prevalence of DQD in a sample size of 190. Infant's age, lifting frequency and hand dominance were proved significant risk factors. However, caregiver's age, history of wrist pain, infant weight and relationship with infant were proved insignificant. Mean PRWE pain and functional scores were 23.14±7.72 and 18.53±6.09, respectively.

Environmental enteric dysfunction (EED) is a subclinical syndrome of altered small intestinal function postulated to be an important contributor to childhood undernutrition. The role of small intestinal bacterial communities in the pathophysiology of EED is poorly defined due to a paucity of studies where there has been a direct collection of small intestinal samples from undernourished children. Sixty-three members of a Pakistani cohort identified as being acutely malnourished between 3 and 6 months of age and whose wasting (weight-for-length Z -score [WLZ]) failed to improve after a 2-month nutritional intervention underwent esophagogastroduodenoscopy (EGD). Paired duodenal luminal aspirates and duodenal mucosal biopsies were obtained from 43 children. Duodenal microbiota composition was characterized by sequencing bacterial 16S rRNA gene amplicons. Levels of bacterial taxa (amplicon sequence variants [ASVs]) were referenced to anthropometric indices, histopathologic severity in biopsies, expression of selected genes in the duodenal mucosa, and fecal levels of an immunoinflammatory biomarker (lipocalin-2). A "core" group of eight bacterial ASVs was present in the duodenal samples of 69% of participants. Streptococcus anginosus was the most prevalent, followed by Streptococcus sp., Gemella haemolysans , Streptococcus australis , Granulicatella elegans , Granulicatella adiacens, and Abiotrophia defectiva . At the time of EGD, none of the core taxa were significantly correlated with WLZ. Statistically significant correlations were documented between the abundances of Granulicatella elegans and Granulicatella adiacens and the expression of duodenal mucosal genes involved in immune responses (dual oxidase maturation factor 2, serum amyloid A, and granzyme H). These results suggest that a potential role for members of the oral microbiota in pathogenesis, notably Streptococcus , Gemella, and Granulicatella species, warrants further investigation.IMPORTANCEUndernutrition among women and children is a pressing global health problem. Environmental enteric dysfunction (EED) is a disease of the small intestine (SI) associated with impaired gut mucosal barrier function and reduced capacity for nutrient absorption. The cause of EED is ill-defined. One emerging hypothesis is that alterations in the SI microbiota contribute to EED. We performed a culture-independent analysis of the SI microbiota of a cohort of Pakistani children with undernutrition who had failed a standard nutritional intervention, underwent upper gastrointestinal tract endoscopy, and had histologic evidence of EED in their duodenal mucosal biopsies. The results revealed a shared group of bacterial taxa in their duodenums whose absolute abundances were correlated with levels of the expression of genes in the duodenal mucosa that are involved in inflammatory responses. A number of these bacterial taxa are more typically found in the oral microbiota, a finding that has potential physiologic and therapeutic implications., Competing Interests: The authors declare no conflict of interest.

Background: The reduction in severe and moderate acute malnutrition (SAM and MAM) rates in Pakistan has been sub-optimal compared to other low-and middle-income countries (LMICs). Specially-formulated products have been designed globally to manage SAM and MAM, such as ready-to-use therapeutic food (RUTF) and ready-to-use supplementary food (RUSF), with variable efficacies. RUTF is primarily produced and patented in industrialized countries, raising supply challenges in resource-constrained regions with a high burden of acute malnutrition. RUSF minimizes costs by using locally-available ingredients while providing similar nutritional value. In this study, we compared the efficacy, side effects, and compliance of two months of supplementation with either RUTF or RUSF., Methods: Children aged nine months in the rural district of Matiari, Pakistan, with a weight-for-height z-score (WHZ) <-2 received either RUTF (500 kcal sachet) for two months in 2015 or RUSF (520 kcal sachet) for two months in 2018., Results: The RUSF group had a higher height gain and mid-upper arm circumferences (MUAC) score. Higher compliance was noted with lower side effects in the RUSF group. A higher compliance rate did correlate with the growth parameters in respective groups., Conclusion: Our study found that both RUTF and RUSF partially improve the anthropometric status of acutely malnourished children, with neither being superior to the other., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Sarfraz et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Background: Meningococcaemia is a serious bacterial disease caused by Neisseria meningitidis resulting in septicaemia and meningitis in previously well individuals and is associated with serious consequences including mortality. As the data from our region is scarce, the study was conducted with an aim to highlight the disease manifestations and outcomes so that appropriate interventions are devised., Method: A descriptive study was conducted in Paediatrics A Unit from 1st March 2020 to 30th September 2021 after approval of institutional review board. Children admitted with features of meningococcaemia in the form of fever and typical petechial purpuric rash were included in the study., Results: A total of 46 patients with meningococcaemia were included in the study. Of these, 24 (52.2%) were male. Majority of the patients 22 (47.8%) were in the age group of 1 year to five years, Major complications were meningitis in 39 (84.8%), septic shock in 26 (56.5%) and purpura fulminans in 12 (26.1%) patients. Mortality was documented in 6 (13%) patients. Outcome was associated with age. (p= 0.039). There was a significant difference in outcome in patients with prolonged PT/APTT (p=0.031), purpura fulminans (p=0.000) and septic shock (p=0.021)., Conclusion: Meningococcaemia is prevalent in our region in paediatric population. The disease has a fulminant course with a myriad of complications and potentially fatal outcomes especially in children under one year of age.

Background: The underperformance of oral vaccines in children of low- and middle-income countries is partly attributable to underlying environmental enteric dysfunction (EED)., Methodology: We conducted a longitudinal, community-based study to evaluate the association of oral rotavirus vaccine (Rotarix®) seroconversion with growth anthropometrics, EED biomarkers and intestinal enteropathogens in Pakistani infants. Children were enrolled between three to six months of their age based on their nutritional status. We measured serum anti-rotavirus immunoglobulin A (IgA) at enrollment and nine months of age with EED biomarkers and intestinal enteropathogens., Results: A total of 391 infants received two doses of rotavirus (RV) vaccine. 331/391 provided paired blood samples. Of these 331 children, 45% seroconverted at 9 months of age, 35% did not seroconvert and 20% were seropositive at baseline. Non-seroconverted children were more likely to be stunted, wasted and underweight at enrollment. In univariate analysis, insulin-like growth factor (IGF) concentration at 6 months were higher in seroconverters, median (25th, 75th percentile): 26.3 (16.5, 43.5) ng/ml vs. 22.5 (13.6, 36.3) ng/ml for non-seroconverters, p-value = 0.024. At nine months, fecal myeloperoxidase (MPO) concentrations were significantly lower in seroconverters, 3050(1250, 7587) ng/ml vs. 4623.3 (2189, 11650) ng/ml in non-seroconverted children, p-value = 0.017. In multivariable logistic regression analysis, alpha-1 acid glycoprotein (AGP) and IGF-1 concentrations were positively associated with seroconversion at six months. The presence of sapovirus and rotavirus in fecal samples at the time of rotavirus administration, was associated with non-seroconversion and seroconversion, respectively., Conclusion: We detected high baseline RV seropositivity and impaired RV vaccine immunogenicity in this high-risk group of children. Healthy growth, serum IGF-1 and AGP, and fecal shedding of rotavirus were positively associated with RV IgA seroconversion following immunization, whereas the presence of sapovirus was more common in non-seroconverters., Trial Registration: Clinical Trials ID: NCT03588013., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The relationship between environmental factors and child health is not well understood in rural Pakistan. This study characterized the environmental factors related to the morbidity of acute respiratory infections (ARIs), diarrhea, and growth using geographical information systems (GIS) technology. Anthropometric, address and disease prevalence data were collected through the SEEM (Study of Environmental Enteropathy and Malnutrition) study in Matiari, Pakistan. Publicly available map data were used to compile coordinates of healthcare facilities. A Pearson correlation coefficient ( r ) was used to calculate the correlation between distance from healthcare facilities and participant growth and morbidity. Other continuous variables influencing these outcomes were analyzed using a random forest regression model. In this study of 416 children, we found that participants living closer to secondary hospitals had a lower prevalence of ARI ( r = 0.154, p < 0.010) and diarrhea (r = 0.228, p < 0.001) as well as participants living closer to Maternal Health Centers (MHCs): ARI ( r = 0.185, p < 0.002) and diarrhea ( r = 0.223, p < 0.001) compared to those living near primary facilities. Our random forest model showed that distance has high variable importance in the context of disease prevalence. Our results indicated that participants closer to more basic healthcare facilities reported a higher prevalence of both diarrhea and ARI than those near more urban facilities, highlighting potential public policy gaps in ameliorating rural health.

Environmental enteric dysfunction (EED) is a subclinical condition of intestinal inflammation, barrier dysfunction and malabsorption associated with growth faltering in children living in poverty. This study explores association of altered duodenal permeability (lactulose, rhamnose and their ratio) with higher burden of enteropathogen in the duodenal aspirate, altered histopathological findings and higher morbidity (diarrhea) that is collectively associated with linear growth faltering in children living in EED endemic setting. In a longitudinal birth cohort, 51 controls (WHZ > 0, HAZ > -1.0) and 63 cases (WHZ< -2.0, refractory to nutritional intervention) were recruited. Anthropometry and morbidity were recorded on monthly bases up to 24 months of age. Dual sugar assay of urine collected after oral administration of lactulose and rhamnose was assessed in 96 children from both the groups. Duodenal histopathology (n = 63) and enteropathogen analysis of aspirate via Taqman array card (n = 60) was assessed in only cases. Giardia was the most frequent pathogen and was associated with raised L:R ratio (p = 0.068). Gastric microscopy was more sensitive than duodenal aspirate in H. pylori detection. Microscopically confirmed H. pylori negatively correlated with HAZ at 24 months (r = -0.313, p = 0.013). Regarding histopathological parameters, goblet cell reduction significantly correlated with decline in dual sugar excretion (p< 0.05). Between cases and controls, there were no significant differences in the median (25th, 75th percentile) of urinary concentrations (μg/ml) of lactulose [27.0 (11.50, 59.50) for cases vs. 38.0 (12.0, 61.0) for controls], rhamnose [66.0 (28.0, 178.0) vs. 86.5 (29.5, 190.5)] and L:R ratio [0.47 (0.24, 0.90) vs. 0.51 (0.31, 0.71)] respectively. In multivariable regression model, 31% of variability in HAZ at 24 months of age among cases and controls was explained by final model including dual sugars. In conclusion, enteropathogen burden is associated with altered histopathological features and intestinal permeability. In cases and controls living in settings of endemic enteropathy, intestinal permeability test may predict linear growth. However, for adoption as a screening tool for EED, further validation is required due to its complex intestinal pathophysiology., Competing Interests: The authors have declared that no competing interests exist.

Background & Aims: Environmental enteric dysfunction (EED) limits the Sustainable Development Goals of improved childhood growth and survival. We applied mucosal genomics to advance our understanding of EED., Methods: The Study of Environmental Enteropathy and Malnutrition (SEEM) followed 416 children from birth to 24 months in a rural district in Pakistan. Biomarkers were measured at 9 months and tested for association with growth at 24 months. The duodenal methylome and transcriptome were determined in 52 undernourished SEEM participants and 42 North American controls and patients with celiac disease., Results: After accounting for growth at study entry, circulating insulin-like growth factor-1 (IGF-1) and ferritin predicted linear growth, whereas leptin correlated with future weight gain. The EED transcriptome exhibited suppression of antioxidant, detoxification, and lipid metabolism genes, and induction of anti-microbial response, interferon, and lymphocyte activation genes. Relative to celiac disease, suppression of antioxidant and detoxification genes and induction of antimicrobial response genes were EED-specific. At the epigenetic level, EED showed hyper-methylation of epithelial metabolism and barrier function genes, and hypo-methylation of immune response and cell proliferation genes. Duodenal coexpression modules showed association between lymphocyte proliferation and epithelial metabolic genes and histologic severity, fecal energy loss, and wasting (weight-for-length/height Z < -2.0). Leptin was associated with expression of epithelial carbohydrate metabolism and stem cell renewal genes. Immune response genes were attenuated by giardia colonization., Conclusions: Children with reduced circulating IGF-1 are more likely to experience stunting. Leptin and a gene signature for lymphocyte activation and dysregulated lipid metabolism are implicated in wasting, suggesting new approaches for EED refractory to nutritional intervention. ClinicalTrials.gov, Number: NCT03588013. (https://clinicaltrials.gov/ct2/show/NCT03588013)., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Background: Stunting affects up to one-third of the children in low-to-middle income countries (LMICs) and has been correlated with decline in cognitive capacity and vaccine immunogenicity. Early identification of infants at risk is critical for early intervention and prevention of morbidity. The aim of this study was to investigate patterns of growth in infants up through 48 months of age to assess whether the growth of infants with stunting eventually improved as well as the potential predictors of growth., Methods: Height-for-age z-scores (HAZ) of children from Matiari (rural site, Pakistan) at birth, 18 months, and 48 months were obtained. Results of serum-based biomarkers collected at 6 and 9 months were recorded. A descriptive analysis of the population was followed by assessment of growth predictors via traditional machine learning random forest models., Results: Of the 107 children who were followed up till 48 months of age, 51% were stunted (HAZ < - 2) at birth which increased to 54% by 48 months of age. Stunting status for the majority of children at 48 months was found to be the same as at 18 months. Most children with large gains started off stunted or severely stunted, while all of those with notably large losses were not stunted at birth. Random forest models identified HAZ at birth as the most important feature in predicting HAZ at 18 months. Of the biomarkers, AGP (Alpha- 1-acid Glycoprotein), CRP (C-Reactive Protein), and IL1 (interleukin-1) were identified as strong subsequent growth predictors across both the classification and regressor models., Conclusion: We demonstrated that children most children with stunting at birth remained stunted at 48 months of age. Value was added for predicting growth outcomes with the use of traditional machine learning random forest models. HAZ at birth was found to be a strong predictor of subsequent growth in infants up through 48 months of age. Biomarkers of systemic inflammation, AGP, CRP, IL1, were also strong predictors of growth outcomes. These findings provide support for continued focus on interventions prenatally, at birth, and early infancy in children at risk for stunting who live in resource-constrained regions of the world.

Objective: To determine the frequency of empty follicle syndrome in in-vitro fertilization-stimulated cycles and to evaluate the causes associated with this phenomenon., Methods: The retrospective study was conducted at the Baqai Institute of Reproduction and Developmental Sciences, Karachi, and comprised data related to period from January 2012 to December 2016. Other than demographic information, laboratory investigations, ultrasound findings and treatment protocols were noted and evaluated. Data was statistically analysed using SPSS 21., Results: Of the 1120 cycles studied, empty follicle syndrome was found in 72 (6.4%). The possible leading causes included advanced age in 64 (88.8%) patients, longer duration of infertility in 56 (77.7%), higher baseline follicle stimulating hormone in 56 (77.7%), and lower oestradiol level before human chorionic gonadotrophin injection in 58 (80%). Empty follicles were significantly correlated with age of the patient, years of infertility, serum follicle stimulating hormone level and stimulation length (p<0.05)., Conclusions: Empty follicle syndrome is a rare occurrence. Screening of patients for risk factors before considering IVF cycles is mandatory. The patients should be counseled for possible EFS even in first cycle.

Objective: To measure impact of social, spiritual, occupational, physical, financial and emotional wellness on intellectual wellness of medical teachers., Methods: The cross-sectional study was conducted at 8 medical universities in the Sindh province of Pakistan from January 2012 to December 2014, and comprised faculty members. Using a self-administered questionnaire, teachers' responses on physical, emotional, social, spiritual, intellectual, occupational and financial spokes of the wellness wheel that were graded on a 4-point Likert scale. Generalised linear models were executed to find the best predictive scores for intellectual wellness., Results: Out of 300 subjects approached, 261(87%) returned the questionnaire fully completed. Of them, 86(33%) were males and 175(67%) were females. Occupational wellness followed by spiritual, financial and emotional wellness depicted significantly high correlation with IW (p<0.05). Adjusted analysis revealed significant effects of spiritual, financial and occupational wellness (p<0.05). Findings of Models 1 and 2 suggested that holding the effect of other wellness subscales, intellectual wellness increased up to 0.485 score and 0.245 score due to occupational and spiritual wellness (p<0.05) respectively, while financial wellness was not significant (p>0.05). In the final model, occupational wellness effect increased to 0.504 score and spiritual wellness effect projected to 0.273 score for intellectual wellness (p<0.05)., Conclusions: Amongst all the spokes of the wellness wheel, occupational, financial, social, emotional and spiritual wellness improved mental capabilities of medical teachers to overcome the challenges for acquiring intellectual wellness. However, wellness at workplace played the major role in enhancing intellectual proficiencies.

Background: A major limitation to understanding the etiopathogenesis of environmental enteric dysfunction (EED) is the lack of a comprehensive, reproducible histologic framework for characterizing the small bowel lesions. We hypothesized that the development of such a system will identify unique histology features for EED, and that some features might correlate with clinical severity., Methods: Duodenal endoscopic biopsies from two cohorts where EED is prevalent (Pakistan, Zambia) and North American children with and without gluten sensitive enteropathy (GSE) were processed for routine hematoxylin & eosin (H&E) staining, and scanned to produce whole slide images (WSIs) which we shared among study pathologists via a secure web browser-based platform. A semi-quantitative scoring index composed of 11 parameters encompassing tissue injury and response patterns commonly observed in routine clinical practice was constructed by three gastrointestinal pathologists, with input from EED experts. The pathologists then read the WSIs using the EED histology index, and inter-observer reliability was assessed. The histology index was further used to identify within- and between-child variations as well as features common across and unique to each cohort, and those that correlated with host phenotype., Results: Eight of the 11 histologic scoring parameters showed useful degrees of variation. The overall concordance across all parameters was 96% weighted agreement, kappa 0.70, and Gwet's AC 0.93. Zambian and Pakistani tissues shared some histologic features with GSE, but most features were distinct, particularly abundance of intraepithelial lymphocytes in the Pakistani cohort, and marked villous destruction and loss of secretory cell lineages in the Zambian cohort., Conclusions: We propose the first EED histology index for interpreting duodenal biopsies. This index should be useful in future clinical and translational studies of this widespread, poorly understood, and highly consequential disorder, which might be caused by multiple contributing processes, in different regions of the world., Competing Interests: I have read the journal's policy and the authors of this manuscript have the following competing interests: Dr. Phillip Tarr is a consultant for Takeda Pharmaceuticals and the Bill & Melinda Gates Foundation. Dr. Phillip Tarr is also a consultant, member of the Scientific Advisory Board, holder of equity, and a co-inventor on a patent that might earn royalties for MediBeacon Inc. The remainder of the authors have declared that no competing interests exist.

Objective: To estimate the phytochemical composition, antioxidant, antifungal, cytotoxic and phytotoxic activities of three important medicinal plants of Poonch valley, Azad Kashmir Pakistan., Methods: Phytochemicals were investigated for both qualitative (phenols, flavonoids, alkaloids and saponins) and quantitative (total phenolic and flavonoid contents) analysis of plant extracts prepared in ethanol, ethyl acetate and benzene. Moreover, antioxidant potential was investigated by screening botanical extracts against 2, 2-diphenyl-1-picrylhydrazyl scavenging assay, phosphomolybdenum and total reducing power assay. While, disc diffusion method was used for determination of antifungal potential of all extracts against four pathogenic strains. Furthermore, brine shrimp cytotoxicity assay was conducted for cytotoxic potential and radish seed germination assay was carried out for estimation of allelopathic potential of plant extracts., Results: Phytochemical evaluation showed the presence of phenolics, flavonoids, alkaloids and saponins in almost all plants. In addition to significant antioxidant potential, Rubia cordifolia (ethanolic extract) revealed highest amount of phenolics and flavonoids. The said plant also exhibited good cytotoxic activity. Among plant extracts highest antifungal activity and allelopathic potential was recorded by Oxalis latifolia (ethyl acetate extract). Ethanol and ethyl acetate were observed to be the best choice of solvents. The best solvents-based results were obtained by Ethanol followed by Ethyl acetate and Benzene., Conclusion: For further studies, Rubia cordifolia and Oxalis latifolia are the most promising medicinal plants of Poonch valley, Azad Kashmir Pakistan at molecular level in order to isolate the compounds concerning with pronounced activities.

Environmental Enteric Dysfunction (EED) is an acquired small intestinal inflammatory condition underlying high rates of stunting in children <5 years of age in low- and middle-income countries. Children with EED are known to have repeated exposures to enteropathogens and environmental toxins that leads to malabsorptive syndrome. We aimed to characterize association of linear growth faltering with enteropathogen burden and subsequent changes in EED biomarkers. In a longitudinal birth cohort (n = 272), monthly anthropometric measurements (Length for Age Z score- LAZ) of asymptomatic children were obtained up to 18 months. Biological samples were collected at 6 and 9 months for the assessment of biomarkers. A customized TaqMan array card was used to target 40 enteropathogens in fecal samples. Linear regression was applied to study the effect of specific enteropathogen infection on change in linear growth (ΔLAZ). Presence of any pathogen in fecal sample correlated with serum flagellin IgA (6 mo, r = 0.19, p = 0.002), fecal Reg 1b (6 mo, r = 0.16, p = 0.01; 9mo, r = 0.16, p = 0.008) and serum Reg 1b (6 mo, r = 0.26, p<0.0001; 9 mo, r = 0.15, p = 0.008). At 6 months, presence of Campylobacter [β (SE) 7751.2 (2608.5), p = 0.003] and ETEC LT [β (SE) 7089.2 (3015.04), p = 0.019] was associated with increase in MPO. Giardia was associated with increase in Reg1b [β (SE) 72.189 (26.394), p = 0.006] and anti-flic IgA[β (SE) 0.054 (0.021), p = 0.0091]. Multiple enteropathogen infections in early life negatively correlated with ΔLAZ, and simultaneous changes in gut inflammatory and permeability markers. A combination vaccine targeting enteropathogens in early life could help in the prevention of future stunting., Competing Interests: The authors have declared that no competing interests exist.

Background: Environmental Enteropathy (EE), characterized by alterations in intestinal structure, function, and immune activation, is believed to be an important contributor to childhood undernutrition and its associated morbidities, including stunting. Half of all global deaths in children < 5 years are attributable to under-nutrition, making the study of EE an area of critical priority., Methods: Community based intervention study, divided into two sub-studies, 1) Longitudinal analyses and 2) Biopsy studies for identification of EE features via omics analyses. Birth cohorts in Matiari, Pakistan established: moderately or severely malnourished (weight for height Z score (WHZ) < - 2) children, and well-nourished (WHZ > 0) children. Blood, urine, and fecal samples, for evaluation of potential biomarkers, will be collected at various time points from all participants (longitudinal analyses). Participants will receive appropriate educational and nutritional interventions; non-responders will undergo further evaluation to determine eligibility for further workup, including upper gastrointestinal endoscopy. Histopathological changes in duodenal biopsies will be compared with duodenal biopsies obtained from USA controls who have celiac disease, Crohn's disease, or who were found to have normal histopathology. RNA-Seq will be employed to characterize mucosal gene expression across groups. Duodenal biopsies, luminal aspirates from the duodenum, and fecal samples will be analyzed to define microbial community composition (omic analyses). The relationship between histopathology, mucosal gene expression, and community configuration will be assessed using a variety of bioinformatic tools to gain better understanding of disease pathogenesis and to identify mechanism-based biomarkers. Ethical review committees at all collaborating institutions have approved this study. All results will be made available to the scientific community., Discussion: Operational and ethical constraints for safely obtaining intestinal biopsies from children in resource-poor settings have led to a paucity of human tissue-based investigations to understand and reverse EE in vulnerable populations. Furthermore, EE biomarkers have rarely been correlated with gold standard histopathological confirmation. The Study of Environmental Enteropathy and Malnutrition (SEEM) is designed to better understand the pathophysiology, predictors, biomarkers, and potential management strategies of EE to inform strategies to eradicate this debilitating pathology and accelerate progress towards the 2030 Sustainable Development Goals., Trial Registration: Retrospectively registered; clinicaltrials.gov ID NCT03588013 .

Despite nutrition interventions, stunting thought to be secondary to underlying environmental enteropathy (EE) remains pervasive among infants residing in resource-poor countries and remains poorly characterized. From a birth cohort of 380 children, 65 malnourished infants received 12 weeks of nutritional supplementation with ready-to-use therapeutic food (RUTF). Eleven children with insufficient response to RUTF underwent upper endoscopy with duodenal biopsies, which were compared with U.S., age-matched specimens for healthy, celiac disease, non-celiac villous atrophy, non-celiac intraepithelial lymphocytosis, and graft-versus-host disease patients. Of the 11 children biopsied, EE was found in 10 (91%) with one subject with celiac disease. Morphometry demonstrated decreased villus-to-crypt (V:C) ratios in EE relative to healthy and non-celiac lymphocytosis patients. Environmental enteropathy villus volumes were significantly decreased relative to healthy controls. In EE, average CD3 + cells per 100 epithelial cells and per 1,000 µm 2 of lamina propria and the number of lamina propria CD20 + B-cell aggregates were increased relative to all other groups. Our results indicate that V:C ratios are reduced in EE but are less severe than in celiac disease. Environmental enteropathy intraepithelial and lamina propria T lymphocytosis is of greater magnitude than that in celiac disease. The increases in lamina propria B and T lymphocytes suggest that non-cytolytic lymphocytic activation may be a more prominent feature of EE relative to celiac disease. These results provide new insights into shared yet distinct histological and immunological features of EE and celiac disease in children.

Background: Environmental Enteric Dysfunction (EED) in children from low-income countries has been linked to linear growth declines. There is a critical need to identify sensitive and early EED biomarkers., Objective: Determine whether levels of antibodies against bacterial components flagellin (flic) and lipopolysaccharide (LPS) predict poor growth., Design/methods: In a prospective birth cohort of 380 children in rural Pakistan blood and stool samples were obtained at ages 6 and 9 months. Linear mixed effects models were used to examine longitudinal associations between quartiles of anti-flic and anti-LPS antibodies and changes in LAZ, WAZ and WLZ scores. Spearman's correlations were measured between anti-flic and anti-LPS immunoglobulins with measures of systemic/enteric inflammation and intestinal regeneration., Results: Anti-LPS IgA correlated significantly with CRP, AGP and Reg1 serum at 6mo and with MPO at 9mo. In multivariate analysis at 6mo of age, higher anti-LPS IgA levels predicted greater declines in LAZ scores over subsequent 18mo (comparing highest to lowest quartile, β (SE) change in LAZ score/year = -0.313 (0.125), p-value = 0.013). Anti-flic Ig A in the two highest quartiles measured at 9mo of age had declines in LAZ of -0.269 (0.126), p = 0.033; and -0.306 (0.129), p = 0.018 respectively, during the subsequent 18mo of life, compared to those in the lowest quartile of anti-flic IgA., Conclusions and Relevance: Elevated anti-flic IgA and anti-LPS IgA antibodies at 6 and 9mo, predict declines in linear growth. Systemic and enteric inflammation correlated with anti-LPS IgA provides mechanistic considerations for potential future interventions.

Environmental Enteric Dysfunction (EED), a syndrome characterized by chronic gut inflammation, contributes towards stunting and poor response to enteric vaccines in children in developing countries. In this study, we evaluated major putative biomarkers of EED using growth faltering as its clinical proxy. Newborns (n = 380) were enrolled and followed till 18 months with monthly anthropometry. Biomarkers associated with gut and systemic inflammation were assessed at 6 and 9 months. Linear mixed effects model was used to determine the associations of these biomarkers with growth faltering between birth and 18 months. Fecal myeloperoxidase (neutrophil activation marker) at 6 months [β = -0.207, p = 0.005], and serum GLP 2 (enterocyte proliferation marker) at 6 and 9 months [6M: β = -0.271, p = 0.035; 9M: β = -0.267, p = 0.045] were associated with decreasing LAZ score. Ferritin at 6 and 9 months was associated with decreasing LAZ score [6M: β = -0.882, p < 0.0001; 9M: β = -0.714, p < 0.0001] and so was CRP [β = -0.451, p = 0.039] and AGP [β = -0.443, p = 0.012] at 9 months. Both gut specific and systemic biomarkers correlated negatively with IGF-1, but only weakly correlated, if at all with each other. We therefore conclude that EED may be contributing directly towards growth faltering, and this pathway is not entirely through the pathway of systemic inflammation.

A cross section, questionnaire based study was carried out from January 2012 till December 2014 to compare intellectual wellness (IW) awareness on the basis of gender in teachers of basic sciences at medical universities of Karachi, Pakistan. Data was collected from 3 public and 5 private medical universities of Karachi, Pakistan. Questionnaire was tailored from "Wellness Wheel" and responses were aggregated for eight questions in the dimension of IW to obtain aggregate IW score. Reliability of the questionnaire was measured using Cronbach's alpha. The average intellectual score was 24.99 ± 3.93 with a minimum score of 8 and maximum 32.The frequency of keeping informed about research updates was significantly higher in males (p = 0.043) that emphasized significantly better IW awareness of male medical teachers involved in teaching of medical sciences in both public and private medical universities of Pakistan.

Objectives: Currently available vacuum devices used to assist women undergoing complicated labour are unsuitable for use in low-resource settings. The objective of this study was to evaluate the safety and feasibility of a new low-cost vacuum device, named Koohi Goth Vacuum Delivery System (KGVDS), designed for use in low-resource settings., Methods: A hospital-based, multicentre, prospective cohort study with no control group was conducted in Karachi, Pakistan. After training, KGVDS devices were made available for use by labour room staff at their discretion when instrumental delivery was indicated. Women to whom KGVDS was applied were followed from the start of labour until discharge. Feasibility was assessed in terms of successful expulsion of the foetal head following application of KGVDS and ease of use ratings. Safety was assessed by observing maternal and newborn post-delivery outcomes prior to discharge., Results: Koohi Goth Vacuum Delivery System was applied to 137 women requiring instrumental delivery, of whom 111 (81%; 95% CI = 74-88%) successfully expelled the foetal head assisted by KGVDS and 103 (75%) stated that they would agree to use KGVDS again. There were no serious maternal or neonatal injuries or infections related to KGVDS use. The mean score for 'ease of use' given by doctors and midwives using the device was 8 of 10., Conclusions: Koohi Goth Vacuum Delivery System was feasible and safe to use for assisting complicated deliveries in low-resource hospitals in this initial evaluation. Our results indicate that this new device may have the potential to improve birth outcomes in settings where most mortality occurs and that further evaluations should be conducted., (© 2014 John Wiley & Sons Ltd.)

Approximately 10-15% of the married couples remain childless. Conventional in vitro fertilisation (IVF) has been the treatment option for most of these cases. However, it is expensive and only available to a small fraction of the infertile population. The Sindh Institute of Reproductive Medicine (SIRM), Karachi, Pakistan, has introduced a new method of IVF for the first time in Pakistan. Intravaginal culture (IVC) is a simple, reliable and cost-effective alternative to IVF. The procedure can be performed in an office set-up with minor capital equipment. Here, we report a case of a subfertile woman treated by IVC, which resulted in successful fertilisation of the eggs (fertilisation rate 71.42%) and embryo development, with an intrauterine pregnancy. Since this is the first successful case of IVC in Pakistan, comparative success rate and take-home baby rate have yet to be established.

Objective: To assess and compare knowledge, attitudes and practices of male and female medical students of Karachi in the context of physical, emotional and spiritual wellness., Methods: The cross-sectional questionnaire-based survey was conducted from January to December 2010, comprising randomly selected 800 male/female medical students in eight medical colleges of Karachi. The responses--'never,' 'sometimes,' 'mostly' and 'always' (numbered 0-4 )--were analysed in terms of frequency, proportion and percentages by Predictive Analysis Software (PASW) version 18. Chi-square test was applied for comparison of categorical aspects of wellness, and results were declared significant at p < 0.05., Results: Of the 800 questionnaires, 736 (92%) fully-completed questionnaires comprised the study universe. According to the survey, 503 (68.37%), 504 (68.45%) and 658 (89.35%) medical students were aware of the concept of physical, emotional and spiritual wellness respectively. Males had greater involvement in physical activity (p < 0.001), while females selected healthy diet (p < 0.03). The dimensions of physical and emotional wellness on an aggregate did not differ significantly in both genders, but knowledge and practices of spiritual wellness in females gave them direction to face episodes of anxiety for which males had to take help from mentors and psychologists (p < 0.034, p < 0.0001)., Conclusion: Spirituality, the key success factor in better wellness management, was found to be greater among female medical students.

Background: Dental caries is highly prevalent and a significant public health problem among children throughout the world. Epidemiological data regarding prevalence of dental caries amongst Pakistani pre-school children is very limited. The objective of this study is to determine the frequency of dental caries among pre-school children of Saddar Town, Karachi, Pakistan and the factors related to caries., Methods: A cross-sectional study of 1000 preschool children was conducted in Saddar town, Karachi. Two-stage cluster sampling was used to select the sample. At first stage, eight clusters were selected randomly from total 11 clusters. In second stage, from the eight selected clusters, preschools were identified and children between 3- to 6-years age group were assessed for dental caries., Results: Caries prevalence was 51% with a mean dmft score being 2.08 (±2.97) of which decayed teeth constituted 1.95. The mean dmft of males was 2.3 (±3.08) and of females was 1.90 (±2.90). The mean dmft of 3, 4, 5 and 6-year olds was 1.65, 2.11, 2.16 and 3.11 respectively. A significant association was found between dental caries and following variables: age group of 4-years (p-value < 0.029, RR = 1.248, 95% Bias corrected CI 0.029-0.437) and 5-years (p-value < 0.009, RR = 1.545, 95% Bias corrected CI 0.047-0.739), presence of dental plaque (p-value < 0.003, RR = 0.744, 95% Bias corrected CI (-0.433)-(-0.169)), poor oral hygiene (p-value < 0.000, RR = 0.661, 95% Bias corrected CI (-0.532)-(-0.284)), as well as consumption of non-sweetened milk (p-value < 0.049, RR = 1.232, 95% Bias corrected CI 0.061-0.367)., Conclusion: Half of the preschoolers had dental caries coupled with a high prevalence of unmet dental treatment needs. Association between caries experience and age of child, consumption of non-sweetened milk, dental plaque and poor oral hygiene had been established.

Objective: To assess stress levels, job satisfaction and working hours of the residents in Medicine and Surgery and to explore a correlation among the three factors., Methods: The questionnaire-based. Cross-sectional study was conducted in 2011 at two tertiary level teaching hospitals in Karachi, Pakistan (Ziauddin University Hospital and Aga Khan University Hospital, Karachi, Pakistan). The study population comprised 176 residents, General Health Questionnaire-12 was used to assess the mental health of the residents and a standardised Job Satisfaction Survey tool was used to assess their work satisfaction., Results: A total of 176 residents participated in the study; 115 (65%) from Medicine, and 61 (35%) from surgery. Of the total residents, 99 (56.3%) were under stress, and there was no significant difference in stress between Medicine and Surgery residents. Besides, 133 (75.6%) residents reported to be satisfied with their jobs. There was a statistically significant difference in job satisfaction between Medicine and Surgery residents (p < 0.001). Mean number of working hours per week reported by Medicine and Surgery residents were 74.82 +/- 15.95 and 92.07 +/- 15.91 respectively (p < 0.001). A positive correlation of mean working hours with both stress (p = 0.009) and job satisfaction (p = 0.029) was found., Conclusions: Medicine and Surgery residents tend to differ on mean working hours and job satisfaction. The greater mean working hours of the residents could be a possible reason for increased stress and decreased job satisfaction among the residents.

Objective: To assess the survival of freezing cleaved human embryos through vitrification., Methods: The prospective study was conducted at the Karachi-based Sindh Institute of Reproductive Medicine between June 2008 and June 2009. The cryopreservation of embryos being a new technology in Pakistan, only 19 couples, picked through convenience sampling, comprised the study population. The couples were treated for infertility by in virto fertilisation (IVF) or intracytoplasmic sperm injection (ICSI); 125 surplus embryos were vitrified. Subsequently, 15 embryos were thawed, and transferred in a controlled cycle. SPSS version 11 was used for statistical analysis., Result: After the surplus embryos were vitrified and subsequently thawed and transferred, the survival of the embryos was assessed by the number of blastomeres that were intact. The overall embryo survival rate was (14/15) 93.33%., Conclusion: Vitrification is a simple procedure that requires less time and is likely to become safer and more cost-effective with time. Survival rate after thawing and preserving is high, but comparative success rates in terms of pregnancy and taking-home-baby rates are yet to be established in Pakistan.

Objective: To analyse the result of intra-cytoplasmic sperm injection (ICSI) by using surgically retrieved sperms in the azoospermic male., Methods: Eight (9 cycles) azoospermic men were given the intra-cytoplasmic sperum injection after sperms were collected through surgical retrieval either by percutaneous epididymal sperm aspiration (PESA) or testicular sperm extraction (TESE) at the Sindh Institute of Reproductive Medicine (SIRM), Karachi, which is an assisted reproductive technology centre. Fertilisation, cleavage and pregnancy rates were calculated in their spouses with surgical sperm collection (SSC) and intra-cytoplasmic sperm injection. The study related to the period between September 2007 and February 2009. Statistical analysis were done using SPSS version 11.0., Result: After the intra-cytoplasmic sperm injection procedure, fertilisation rate of 72.72% and cleavage rate of 78.12% were achieved in the eight patients. A total of 3 (33.33%) clinical pregnancies were obtained through the transfer of embryo from surgical sperm collection in the azoospermic men., Conclusion: Intra-cytoplasmic sperm injection with surgical sperm collection proved to be the only successful treatment for men with azoospermia. It gave 33.33% chances of fatherhood in men who were previously labelled infertile. Results were found to be promising and are expected to improve with time and experience.

Objective: To compare the role of new approaches and methods in the development of understanding and learning capabilities of undergraduate medical students in the module of respiratory physiology., Methods: A comparative cross sectional study was conducted in Bahria Univiersity Medical & Dental College using a sample size of 198 students from February till May 2010. Difficulties in understanding the concepts of respiratory physiology was analyzed in groups A (Batch 2008-2013) and assessed through test performance. In group B (Batch 2009-2014) "integrated learner-and teacher-centered" approach was implemented by introduction of problem based learning, case based sessions, students seminar, quizzes and structured assignments. Response of group B students was evaluated in terms of concepts and performance by identical feedback form and assessment test. The data was analyzed using SPSS version 15.0 and applying the Chi-square test., Results: The mean age of Karachi students of Groups A and B was 20 +/- 2 years. Statistically significant improvement in conceptual understanding of Group B (84%) was noticed compared to Group A (42%). Conceptual understanding of Group B (84%) was observed, as compared to Group A (42%), Conclusion: Supplementation of teaching methodologies with augmented interaction improved significant learning, performance and satisfaction among group B students.

Background: Both hepatitis B and C are common infections affecting masses and are leading causes of Chronic Liver Disease in Pakistan as well as worldwide. In majority of cases both viral diseases spread by factors that are preventable. The present study is conducted to determine the identifiable risk factors in patients admitted with Chronic Hepatitis B and C., Methods: An observational study was carried out for a period of 6 months. All age groups and both sexes were included. The patients were interviewed and the identifiable risk factors were looked for. The standard methods for detection of Hepatitis B and C were used., Results: One-hundred and ten patients were studied from January to July 2009. Sixty-five patients had Hepatitis C, 35 had Hepatitis B, and 10 had both Hepatitis B and C. Ninety-three patients had a history of injections and transfusions etc., and 38 had surgical scars. Tattoos were present in 42 patients and nose and/or ear piercing marks were present in 28 patients. The number of risk factors increased in co-infection., Conclusion: There is a role of unhygienic health delivery practices, lack of awareness and resources for standard screening protocol for spread of Hepatitis B and C.

Objective: To determine the frequency of high-risk subtypes of Human papillomavirus (HPV) 16, 18, 31 and 45 in squamous cell carcinoma of cervix (SCC) among women., Methods: A descriptive study was conducted in the department of Molecular Genetics Laboratory, Ziauddin University Hospital, North Nazimabad, Karachi, Pakistan. A total of fifty formalin-fixed paraffin embedded samples (blocks) of SCC of cervix were collected from two Pathology laboratories through convenience sampling. These blocks were analyzed for presence of HPV and its subtype. DNA was extracted by QIAamp DNA Kit and amplification was done by Polymerase chain reaction (PCR). General primers were used for HPV DNA and HPV genotyping for 16, 18, 31 and 45 was done by using a standard kit., Results: HPV was detected in nine (18%) out of fifty paraffin embedded tissues of squamous cell carcinoma of cervix. Out of the 9 cases that were positive, five (55.6%) were infected with HPV16, while in the rest of the positive samples, the genotype could not be identified in four (44.4%). HPV was not determined in majority (82%) of the samples., Conclusion: This study concluded that HPV might not be the major cause of SCC of cervix. There could be other causes among women leading to prevalentHPV types that the study did not look into due to limitations.

Objective: To determine the effect of oral Misoprostol in labour induction with respect to ease of administration and induction-to-delivery time interval., Study Design: Observational study., Place and Duration of Study: Gynaecology/Obstetrics Department, Holy Family Hospital, Rawalpindi, from March to August 2006., Methodology: Women with live singleton pregnancy of > 37 weeks gestation with cephalic presentation, with an indication for induction of labour were inducted. Oral misoprostol 50 microg to 400 microg was given in divided doses at 4 hours interval upto a maximum of 4 doses, till labour was induced. Fetomaternal outcome and induction to delivering time interval in hours was noted., Results: In 6 months duration, 250 mothers were recruited for the study. The main indication for labour induction was post date pregnancy (52%) and oxytocin was given in 50% cases. The majority (96%) of mothers went into labour but 4% (9) had failed induction. The majority (73%, n=176/241) of mothers delivered vaginally, 99% being delivered in the first 24 hours. Mean induction-delivery interval was 11+/-2.7 hours. Sixty five (27%) had to undergo emergency lower segment caesarean section, the major indication being fetal distress (41.5%, n=27) and meconium staining of liquor (40%, n=26). Again, a majority (95%) of the babies were delivered with good Apgar score. However, 10.8% developed meconium aspiration syndrome. Early neonatal deaths occurred in 0.8% (2) cases. Maternal hyper stimulation was seen in 1 case (0.4%)., Conclusion: Oral misoprostol as an agent for labour induction in term pregnancy was easy to administer and the majority of women (99%) delivered in the first 24 hours.

Objective: To determine the frequency of hypothyroidism in patients suffering from homozygous beta-thalassaemia., Methods: This descriptive study included 70 diagnosed thalassaemia major patients aged 5-14 years. Demographic data as well as history of blood transfusion and chelation therapy was collected. Random blood samples were drawn and thyroid profile (serum thyroxine [T4], triiodothyronine [T3] and thyroid stimulating hormone concentrations [TSH]) was done by enzyme-linked immunosorbent assay (ELISA). Primary hypothyroidism was defined by a TSH level > 4 microIU/ml. Results were analysed by descriptive statistical methods., Results: Primary hypothyroidism was seen in 18 (25.7%) patients. Of these, 17 had normal T4 levels with elevated TSH levels consistent with a diagnosis of compensated primary hypothyroidism whereas only one patient showed a decreased T4 level with elevated TSH (uncompensated primary hypothyroidism). Mean age of hypothyroid patients was 9.2 +/- 2.6 years. Frequency of hypothyroidism was associated with increased serum ferritin levels., Conclusion: Primary hypothyroidism occurs in a significant proportion of thalassaemia major patients in the absence of obvious clinical signs of hypothyroidism. Regular follow-up for early detection and timely treatment of such complications could improve the quality of life of these patients.

Objective: To compare body mass index (BMI), waist circumference (WC), and body fat percentage (%BF), as index of overweight and obesity in young adults. We also intended to find an association between lifestyle behaviours and obesity., Methods: A cross-sectional study was conducted at Fatima Jinnah Dental College, Karachi, during 2007 to 2008, with 192 first year dental students, (18-21years) of high socioeconomic class. All were questioned regarding lifestyle behaviours. Overweight and obesity were estimated by measuring %BF, BMI, and WC. For %BF, skinfold thickness was measured using skinfold calipers. BMI > or = 23.0-24.9 kg/m2 was taken as overweight and > or = 25.0 kg/m2 as obese (Asians criteria proposed by Western Pacific Regional Office of World Health Organization). WC using Asian cutoff values for overweight and obesity were: males > or = 78 cm and > or = 90 cm; females > or = 72 cm and > or = 80 cm, respectively. Body fat percentage used to define overweight and obesity was: males 22.1-27.0 and > 27.1; females 27.1-32.0 and > 32.1, respectively. Pearson's correlation was done between the BMI, WC and %BF with statistical significance taken at P < 0.01., Results: BMI (Expressed as mean +/- SD) in males and females was 23.82 +/- 3.88 and 20.98 +/- 4.12 respectively. WC was 83.63 +/- 10.20 cm in males and 70.22 +/- 9.36 cm in females. %BF was 22.32 +/- 6.27 in males and 28.73 +/- 6.65 in females, with an overall 60.8% females and 44.4% males found to be overweight or obese. Obesity was underpredicted by BMI when compared to skinfold calipers method. The obese were seen to skip breakfast more often [odds ratio (OR): 2.39], take frequent snacks (OR: 1.58), watch television more (OR: 1.58), and were physically less active than their non-obese counterparts., Conclusion: Body fat percentage using skinfold caliper is a reliable index of obesity. Lack of sleep and skipping of breakfast, are prominent promoters of obesity, in addition to other lifestyle behaviours.

Objective: To determine the relationship between anthropometric variables with blood glucose and lipid concentrations in different stages of hypertension and evaluate the risk factors involved in progression of pre-hypertension stage in to clinical hypertension stage-1., Study Design: Case-control study., Place and Duration of Study: National Institute of Cardiovascular Disease and General Practitioner's Clinics, Karachi, from 2005 to 2007., Methodology: The study was conducted on 276 participants aged 25-65 years and classified into four categories of control, prehypertension (pre-HTN), hypertension (HTN) stage I and II, according to 7th JNC report. Anthropometric measurements and blood chemistry were obtained. The relationship between obesity indices (Body Mass Index [BMI], Waist Circumference [WC], Waist Hip Ratio [WHpR], Waist Height Ratio [WHtR], Basal Metabolic Rate [BMR] and Cardiovascular Disease [CVD] risk factors [blood pressure, serum lipids and glucose levels]) were investigated. The analysis of variance and test of Pearson's coefficient correlation were applied to assess relationship of all the variables., Results: The mean WHR had significant correlation with both systolic and diastolic BP in all groups of HTN, as well as with blood glucose level in control and pre-HTN groups. BMI was positively correlated with cholesterol (r=0.29) and triglyceride (r=0.25) in HTN stage-1. Impaired and high blood glucose levels were found in pre-HTN group; all lipid parameters except triglycerides were higher in HTN stage I., Conclusion: WHR was positively and independently related to occurrence of arterial hypertension. Elevated levels of WHR, BMI, fasting blood glucose, total cholesterol and triglycerides were responsible for progression of pre-HTN stage in to HTN stage-1.

Human Papilloma Virus (HPV) associated cervical cancer is amongst the ten most common cancers in women in the developing world. New vaccines markedly reduce the economic burden of diseases on a country's health system. Two types of HPV vaccines are available; prophylactic and therapeutic. Prophylactic vaccines target (Late protein) L1 and to a lesser extent L2, which stimulate neutralizing antibody response and are recommended to individuals prior to infection. Therapeutic vaccines are recommended to individuals who are already exposed to HPV infection and these induce a specific T-cell mediated immune response targeted to (Early protein) E6 and E7 oncogenes for the eradication of existing lesion. Several studies have been done which show that these vaccines are highly effective. This review highlights the pathogenesis of HPV infection, importance of HPV vaccines, recent advances and few clinical trials on prophylactic and therapeutic HPV vaccine, their economic impact and the need to implement the vaccination programmes in the developing world.

Background: Due to lack of awareness among health professionals, lymphoma is often misdiagnosed. This study was done to evaluate the clinical features and histopathologic subtypes of lymphoma., Methods: Sixty diagnosed cases of lymphoma were selected (aged 12-65 years) from medical units of Civil Hospital Karachi, during 1993 to 1998. Clinical history, physical examination and basic laboratory investigations including imaging procedures were done in all the patients. The diagnosis of lymphoma was based on histology, following the International Working Formulation classification system. This included lymph node biopsy and in some cases, biopsy of the bone marrow. The Ann Arbor Staging Classification was used to classify the extent of disease., Results: Out of 60 cases of lymphoma, 81.6% (49 cases) were diagnosed as non-Hodgkin's lymphoma and 18.3% (11 cases) as Hodgkin's disease, with an overall male predominance. Both categories exhibited a bimodal age distribution. Lymphadenopathy was the commonest presenting features in both the types of lymphomas; however, patients with Hodgkin's disease had a prominence of 'B' symptoms, whereas abdominal signs and symptoms were more common in non-Hodgkin's lymphoma. On histopathology, majority of non-Hodgkin's lymphomas (91.8%) showed a diffuse pattern, while mixed cellularity was the commonest type seen in Hodgkin's disease (81.8%)., Conclusion: Non-Hodgkin's lymphoma was 4 times more common than Hodgkin's disease. The vast clinical spectrum of lymphoma sometimes delays its diagnosis, leading to its eventual presentation in late stages. A general awareness is hence required among the health professionals regarding its varied clinical presentations.

Objective: To determine the cardiovascular risk factors in users of second generation contraceptives by recording changes in body mass index, blood pressure and electrocardiogram., Design: Cross-sectional study., Place and Duration of Study: The National Institute of Fertility Research Centers at Jinnah Postgraduate Medical Center and PIB Maternity Home Karachi, from July 1997 to 1999., Materials and Methods: Sixty four women volunteered for this study (age range 20-35 years), belonging to low-income group with similar socio-cultural background. The Body Mass Index (BMI) was calculated by measuring height and weight of the subjects; systolic and diastolic blood pressure and ECG recording by standard method. The group means, standard deviations and coefficient correlation for interrelationship among variables in respective groups of subjects were calculated using relevant statistical method and software program., Results: There was no significant difference between BMI of two types of contraceptive users as compared to non users; but BMI was significantly correlated with both systolic and diastolic blood pressures in injectable users as compared to controls. ECG alterations frequently observed in contraceptive users (40%) as compared to controls were normal findings., Conclusion: It was observed that women aged < 30 years and using contraceptives for more than three years had a tendency to gain weight and developed a mild increase in systolic and diastolic blood pressures.

Search for a cure for type-1 diabetes mellitus has lead to many avenues of research, all having the same objective: to replace the lost beta cells and prevent their further destruction by the immune system. Transplantation of islets of Langerhans seems closer to achieving this goal with the recent introduction of new improved immunosuppressive protocols including monoclonal antibodies against the T-lymphocytes. But the need for acquiring beta cells in large numbers rather limits this approach. With the recent advancement in stem cell technology, it may be possible to gather enough stem cells for transplantation purposes. In this regard, embryonic stem cells have shown the greatest promise due to their capacity for unlimited proliferation and differentiation into any cell type. This review discusses the current direction of research regarding diabetes mellitus type-1, while explaining the progress being made in stem cell usage in finding a cure for the disease.

Objective: To note the association of Helicobacter pylori in patients having carcinoma of stomach., Methods: A descriptive study was carried out at the Department of Histopathology, Ziauddin Medical University, Karachi from April 1992 to May 1998. Histological evaluation of 50 cases of carcinoma of stomach was compared with 50 cases each of chronic gastritis and histological normal gastric mucosa. Only those cases of carcinoma of stomach were included that contained sufficient non-neoplastic mucosa in addition to tumour tissue. Three glass slides with serial sections of each case of carcinoma of stomach, chronic gastritis and normal gastric mucosa were freshly cut and stained with H&E, PAS and Giemsa stains. All slides were examined by light microscopy., Results: Helicobacter pylori were identified in 35 cases (70%) of carcinoma of stomach, in 42 cases (84%) of chronic gastritis, and in 12 cases (24%) of normal gastric mucosa. The presence of H. pylori in cases of carcinoma of stomach and chronic gastritis was highly significant (P < 0.001) as compared to normal gastric mucosa. Chronic gastritis was observed in the non-neoplastic mucosa in 48 cases (96%) with carcinoma of stomach. Of 50 cases with carcinoma of the stomach, intestinal type of carcinoma was found in 30 cases (70%), and diffuse type in 15 cases (30%). No significant difference was noted in the prevalence of H. pylori between intestinal type (69%) and diffuse type (71%) gastric carcinoma. Significant Helicobacter pylori associated chronic gastritis was observed in intestinal type (94%) and diffuse type (100%) of gastric carcinoma. The prevalence of H. pylori was insignificant in the presence or absence of mucosal atrophy and intestinal metaplasia in both types of gastric carcinoma., Conclusion: A significant number of H. pylori were found in patients of carcinoma of stomach. Both intestinal and diffuse types of gastric carcinoma showed strong association with H. pylori. Chronic gastritis appears to be the background lesion while atrophy and intestinal metaplasia indicate long term infection.

Of the 43 Aeromonas spp. isolated from various clinical samples 94% isolates were Beta-lactamase producers. The isolates were tested for sensitivity by disc diffusion method which is commonly used in Pakistan. MIC was determined by using Epsilometer test (E-test) method. More than 80% isolates were sensitive to cephalosporins and quinolones. However, resistance to commonly used antibiotics was very high, 94% isolates were resistant to ampicillin which corresponds to the betalactamase production. More than 60% of the isolates were resistant to cotrimoxazole and 40% to chloramphenicol, hence quinolones and cephalosporins appear to be the drugs of choice for treating serious Aeromonas infections. The MIC range of Aeromonas was best for cefotaxime < 0.06 - 1.0 ug/ml. MIC 90 for cefotaxime was 0.50 ug/ml, for imipenem 0.25 ug/ml and for ciprofloxacin 2.0 ug/ml.

Aim and Background: Two forms of hypochromic microcytic anaemia i.e. iron deficiency and beta-thalassaemia trait are common in our society. This study reports the prevalence of iron deficiency anaemia and beta-thalassaemia trait and predictive value of MCV/RBC count ratio to discriminate between two., Methods: Venous blood was taken from 299 students of Karachi Medical & Dental College and Ziauddin Medical University in Na2 EDTA and analyzed by semi-automated Sysmex K-1000 haematology analyzer. MCV/RBC count ratio was used to discriminate between iron deficiency and beta-thalassaemia trait and > 14% was marked as iron deficiency. Hb electrophoresis was used as gold standard test for confirmation. Serum iron and TIBC was performed to confirm iron deficiency anaemia., Results: Iron deficiency was found in 9% while beta-thalassaemia was seen in 3% students. MCV/RBC count ratio showed a positive predictive value of 91%., Conclusions: In areas where iron deficiency anaemia and beta-thalassaemia trait are common, MCV/RBC count ratio can be used to screen out beta-thalassaemia trait.

Most laboratories in Pakistan use expensive imported clinical chemistry reagent kits resulting in high cost/test to the patients. To reduce these costs, reagents were prepared from basic chemicals, substrates and enzymes imported from Sigma Chemical Company U.K. This reduced the cost/test by up to 500% in some reagents. The quality of these reagents was tested by Wellcome External, Q.C. Locally prepared reagents were comparable to or better than commercial reagents systems in terms of accuracy and precision. This paper describes the preparations according to I.F.C.C., costs and quality control of some of the reagents i.e., glucose, calcium, bilirubin, albumin, total protein, urea, ALT, AST and LDH and their comparisons with equivalent commercial kits.