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Culture-independent diagnostics have revealed a larger burden of Shigella among children in low-resource settings than previously recognized. We further characterized the epidemiology of Shigella in the first two years of life in a multisite birth cohort. We tested 41,405 diarrheal and monthly non-diarrheal stools from 1,715 children for Shigella by quantitative PCR. To assess risk factors, clinical factors related to age and culture positivity, and associations with inflammatory biomarkers, we used log-binomial regression with generalized estimating equations. The prevalence of Shigella varied from 4.9%-17.8% in non-diarrheal stools across sites, and the incidence of Shigella-attributable diarrhea was 31.8 cases (95% CI: 29.6, 34.2) per 100 child-years. The sensitivity of culture compared to qPCR was 6.6% and increased to 27.8% in Shigella-attributable dysentery. Shigella diarrhea episodes were more likely to be severe and less likely to be culture positive in younger children. Older age (RR: 1.75, 95% CI: 1.70, 1.81 per 6-month increase in age), unimproved sanitation (RR: 1.15, 95% CI: 1.03, 1.29), low maternal education (<10 years, RR: 1.14, 95% CI: 1.03, 1.26), initiating complementary foods before 3 months (RR: 1.10, 95% CI: 1.01, 1.20), and malnutrition (RR: 0.91, 95% CI: 0.88, 0.95 per unit increase in weight-for-age z-score) were risk factors for Shigella. There was a linear dose-response between Shigella quantity and myeloperoxidase concentrations. The burden of Shigella varied widely across sites, but uniformly increased through the second year of life and was associated with intestinal inflammation. Culture missed most clinically relevant cases of severe diarrhea and dysentery.

A gut microbiome-targeted diet can potentially mitigate chronic diseases, including malnutrition. In a prospective 12-week intervention trial, we evaluated the effects of six different plant-based fermented pickles (~ 50 g/day) on clinical, inflammatory, and gut microbiota parameters in a cohort of women ( n  = 230) in a rural setting with a high prevalence of undernutrition. Blood was collected at two, whereas stool was collected at three timepoints. Among fecal biomarkers, myeloperoxidase (MPO), Lipocalin-2 (LCN2), and 16S rRNA gene sequencing were measured at baseline, 8 th, and 12 th weeks. Overall, the compliance rate was > 70%. Among radish group, WBCs ( p  = 0.002, two-tailed paired T-test) decreased, whereas neutrophils and platelets decreased among both radish ( p  = 0.016, p  = 0.017, two-tailed paired T-test) and carrot ( p  = 0.005, p  = 0.006, two-tailed paired T-test) groups after intervention. Similarly, in lemon-chilli group, platelets decreased while mean corpuscular hemoglobin (MCH) increased ( p  < 0.001, p  = 0.022, two-tailed paired T-test). In onion and lemon-chilli groups, α- ( р  =0.001 and p  = 0.0005, Kruskal-Wallis Test, respectively) and β-diversity indices ( p  = 9e-04 and p  = 0.022, pairwise PERMANOVA, respectively) were significantly increased, post-intervention. Linear discriminant analysis (LDA) of lemon-chilli group identified 25 bacterial taxa markers in 8 th and 12 th week, which included Eggerthellaceae and Oscillospiraceae , Erysipelatoclostridiaceae and Subdoligranulum . Correlation analysis revealed six taxa negatively associated with inflammatory markers such as C-reactive protein (CRP), LCN2, and platelets. Our study provides preliminary evidence that the consumption of traditional fermented pickles leads to beneficial changes in women’s hematological and gut microbiota profiles.

Background Evidence from low- and middle-income countries (LMIC) suggests that home environment is associated with early childhood development outcomes. However, studies from LMIC that have examined how the home environment during middle childhood is associated with intellectual abilities are scarce. The objective of the study was to explore the association between different aspects of the home environment at 7–8 years and fluid reasoning abilities in a rural, high-risk cohort in Pakistan. Methods We employed a cross-sectional research design to examine 1172 children between 7 and 8 years and their families, utilizing the Home Observation for Measurement of Environment for Middle Childhood (HOME-MC) to evaluate various aspects of the home environment and the Fluid Reasoning Index (FRI) of the Wechsler Intelligence for Children (WISC)-5th edition to assess the fluid reasoning abilities of the children. Multivariable regression analyses were used to examine the association between different components of HOME-MC (scored as indices) and FRI scores. Findings Learning materials and opportunities (β = 1.74, 95% CI = 1.15, 2.33) and Responsivity (β = 1.73, 95% CI = 1.07, 2.38) indices had the strongest association with FRI score followed by Family companionship index (β = 1.27, 95% CI = 0.63, 1.90). The eight different indices of the HOME-DC explained 22% of the total variation in FRI scores. Conclusion We conclude that concurrent learning opportunities, parental responsivity and family companionship at home are associated with fluid reasoning abilities during middle childhood which is comparable to what has been found in early childhood years.

The United Nation's Sustainable Development Goals encompass lifelong learning from birth to youth to adulthood (Goal 4) and economic opportunities for young people (Goal 8). The targets include improving access to quality early childhood care and education (ECCE) as well as learning and training opportunities for adolescents and youth. Cross-generational models for young children and youth may offer opportunities to address the interconnections between goals and targets for the next generation. We investigated whether an ECCE programme for young children (3.5-6.5 years) delivered by female youth (18-24 years) in rural Pakistan would be effective on children's school readiness. In partnership with the National Commission for Human Development in Pakistan, we implemented the 'Youth Leaders for Early Childhood Assuring Children are Prepared for School' (LEAPS) programme to train female youth to deliver ECCE. The effectiveness of the LEAPS programme on children's school readiness was evaluated in a cluster-randomised controlled trial. We randomly allocated five clusters (villages) to receive the intervention (n = 170 children) and five clusters to control (n = 170 children). Children's school readiness was assessed after nine months of intervention exposure using the International Development and Early Learning Assessment tool. Analyses was by intention-to-treat. The trial is registered with ClinicalTrials.gov, number NCT02645162. At endline, the intervention group had significantly higher school readiness scores (n = 166, mean percentage score 59.4, 95% CI 52.7 to 66.2) compared with the control group (n = 168, mean percentage score 45.5, 95% CI 38.8 to 52.3). The effect size (Cohen's d) was 0.3. Trained female youth delivered an ECCE programme that was effective in benefitting young children's school readiness. The cross-generational model is a promising approach to support early child development; however, further evaluation of the model is needed to assess the specific benefits to youth including their skills and economic development.

Background The first 1,000 days of life, spanning pregnancy to two years, are critical for development. We investigated the association of maternal, socioeconomic, environmental, nutritional, and childhood characteristics with cognitive, motor, and language development at two years of age. Methods The AMANHI-ACT study enrolled pregnant women between 8 and 19 weeks of gestation from Bangladesh, Pakistan, and Tanzania. Data was collected on sociodemographic factors, maternal BMI and co-morbidities during pregnancy, and weight and gestational age at birth. Child characteristics included vaccination history, infections, and physical growth at predefined intervals up to two years. Child development at two years was assessed using the Bayley Scales of Infant and Toddler Development Third Edition (Bayley-III). The home environment was assessed using the Home Observation for Measurement of the Environment Infant-Toddler (HOME-IT) tool. We performed linear regression and reported site-adjusted regression coefficients with 95% confidence intervals. Results We assessed 3,062 children at two years of age to identify factors associated with their cognitive, language, and motor development scores. Cognitive scores were significantly lower among children whose mothers had no formal education (Adjusted coefficient = -3.8; 95% CI: -5.8, -1.8) and those who had not received any vaccinations (Adjusted coefficient = -6.0; 95% CI: -9.3, -2.6). Language development was similarly affected: children with mothers who had no education (Adjusted coefficient = -4.7; 95% CI: -6.8, -2.5) or only primary-level education (Adjusted coefficient = -4.5; 95% CI: -6.5, -2.5) showed markedly lower scores. Language scores were also reduced in children whose fathers had no education (Adjusted coefficient = -2.4; 95% CI: -4.2, -0.5) or only primary education (Adjusted coefficient = -2.4; 95% CI: -4.1, -0.6). For motor development, children who were stunted (Adjusted coefficient = -2.2; 95% CI: -2.9, -1.4), wasted (Adjusted coefficient = -2.8; 95% CI: -4.3, -1.4), or unvaccinated (Adjusted coefficient = -4.7; 95% CI: -8.0, -1.4) at two years exhibited significantly lower scores. Conclusion Preterm births, vaccination history, parental education, and physical growth significantly affected the first 1000 days of child development, thus identifying areas for targeted intervention to optimize long-term health outcomes in LMICs.

Objective. To assess the effect of vitamin D supplementation on neuropathy specific quality of life (NeuroQoL) in patients with painful diabetic neuropathy. Methods. This prospective, open label study was conducted between June 2012 and April 2013. Patients with symptomatic diabetic neuropathy were given a single dose of 600,000 IU intramuscular vitamin D, and NeuroQol was assessed at baseline and at five follow-up visits every 4 weeks. Results. Of 143 participants, 41.3% were vitamin D deficient (vitamin D < 20 ng/ml). Treatment with vitamin D resulted in a significant increase in 25(OH)D (P<0.0001) and a significant improvement in the NeuroQoL subscale score for emotional distress (P=0.04), with no significant change in the other NeuroQoL domains of painful symptoms and paresthesia, loss of temperature and touch sensation, unsteadiness, limitation in daily activities, and interpersonal problems. There was a significant reduction in patient perception about foot problems on QoL of “quite a lot” (P<0.05) and “very much” (P<0.0001) with a significant reduction in the baseline response of having a “poor” QoL from 5.2% to 0.7% (P<0.0001) and an increase in the response of an “excellent QoL” from 1.5% to 7.4% (P<0.0001). Conclusion. Vitamin D is effective in improving quality of life in patients with painful diabetic neuropathy.

Background Suboptimal feeding practices have a negative impact on children’s health and growth in the first 2 years of life and increase their risk of undernutrition, morbidity, and mortality. The aim of the study was to assess the factors that influence infant and young child feeding practices among urban mothers in a hospital setting at Karachi, Pakistan. Methods A longitudinal multi-center cohort study was conducted in four countries, MULTICENTER BODY COMPOSITION REFERENCE STUDY (MBCRS) to produce normal body composition reference data in healthy infants from 3 months to 24 months of age. Repeated anthropometric (weight, length and head circumference) and body composition measurements using “deuterium dilution method” along with 24-h dietary recall questionnaires were performed on 250 healthy term infants at 3, 6, 9, 12, 18, and 24 months of age. The 24-h dietary recall data from this study was used to assess the breastfeeding and complementary feeding practices in children aged 6–24 months. Results A total of 250 healthy infants were enrolled in the study. A majority of newborns (75.4%) were exclusively breastfed till 3 months of age; however, by 6 months of age, only 30.2% of infants were exclusively breastfed. Only 44.1% of children aged 6–24 months achieved minimum dietary diversity (MDD), 84.7% achieved minimum meal frequency (MMF), and 44.1% achieved a minimum acceptable diet (MAD). 71.4% achieved MDD and MAD and 100% achieved MMF at 24 months. The bivariate analysis found that breastfed children (OR 3.93, 95% CI 2.72–5.68), with employed mothers (OR 1.55, 95% CI 1.06–2.27) who had graduated from secondary school (OR 1.45, 95% CI 1.08–1.94) were more likely to meet minimum dietary diversity. The multivariable analysis showed that only the child’s age was significantly associated with MDD ( p value< 0.0001), with the likelihood of meeting MDD increasing as the children aged; 9 months (OR 18.96, 95% CI 6.63–54.19), 12 months (OR 40.25, 95% CI 14.14–114.58), 18 months (OR 90.02, 95% CI 30.84–262.77) and 24 months (OR 82.14, 95% CI 27.23–247.83). Conclusion Our study revealed that Infant and young child feeding practices are significantly associated with maternal education, employment, and the child’s age. Therefore, it is essential that investments be made towards protective breastfeeding and complementary feeding policies and legislations, emphasis on female education and ensuring the availability of affordable nutritious and diverse foods.

Pakistan is one of the few countries where poliovirus transmission still persists, despite intensive efforts to eradicate the disease. Adequate vaccination coverage is essential to achieve polio eradication, but misconceptions about polio vaccines have hindered vaccination efforts. To address this issue, we conducted a mixed-methods study to explore knowledge and perceptions regarding polio disease and immunization in high-risk areas of Pakistan. We collected quantitative data from 3780, 1258, and 2100 households in Karachi, Bajaur, and Pishin, respectively, and supplemented this with qualitative data from focus group discussions and in-depth interviews. Our findings reveal a high level of awareness about polio and its immunization; however, misperceptions about the polio vaccine persist, leading to refusal for both polio vaccines and routine immunizations. Our study provides up-to-date data on knowledge and perceptions of polio and its immunization and identifies critical gaps. These findings can inform the development of future strategies and innovative approaches to improve the success of the polio program in Pakistan.

Background Globally, malnutrition accounts for at least half of all childhood deaths. Managing malnutrition in the community settings involves identifying malnutrition using a universally validated screening tool and implementing appropriate interventions according to the degree of malnutrition. The aim of this study was to estimate prevalence and associated factors that result in malnutrition among children under-five years of age in Thatta and Sujawal districts in Sindh province, Pakistan. Methods A cross-sectional survey was conducted between May and August 2014. A total of 3964 children under-five years were enrolled in the study. The WHO growth standards height-for-age Z-scores (HAZ), weight-for-height Z-scores (WHZ) and weight-for-age Z-scores (WAZ) were used to measure stunting, wasting and underweight. A structured questionnaire was used to collect data on socioeconomic conditions, family size, maternal education, parity and child morbidity. Multivariable logistic regression was used to determine the risk factors for malnutrition. Results The prevalence of stunting, wasting and underweight were 48.2% (95% CI: 47.1–50.3), 16.2% (95% CI: 15.5–17.9), and 39.5% (95% CI: 38.4–41.5), respectively. Stunting was slightly higher (51%) in boys than in girls (45%) ( p  < 0.001). The proportion of wasting ( p  = 0.039) and underweight ( p  = 0.206) was not significantly different between boys and girls. Fifty percent children in the poorest households were stunted as compared to 42% in the wealthiest households. Children in the poorest households were two times more likely to be wasted (20.6%) than children in the wealthiest households (10.3%) (OR 2.33, CI 1.69–3.21, p  < 0.001). A similar relationship was observed between household wealth and underweight in children (43.8% in poorest and 28.8% in wealthiest households (OR 2.18, CI 1.72–2.77, p  < 0.001). Household wealth was significantly associated with stunting, wasting and underweight. Diarrhea was associated with underweight. Factors such as mother’s education, parity and family size were not associated with malnutrition in our study area. Conclusions The findings of our study revealed that malnutrition was widespread among the children under-five years of age. The food/nutrient based interventions together with improved hygiene practices and household wealth should be targeted to improve malnutrition situation in the study area and in the country.

BackgroundVitamin D deficiency during pregnancy is a public health problem in Pakistan and is prevalent among most women of reproductive age in the country. Vitamin D supplementation during pregnancy is suggested to prevent adverse pregnancy outcomes and vitamin D deficiency in both the mother and her newborn.MethodsWe conducted a double-blinded, randomised controlled trial in Karachi, Pakistan to evaluate the effect of different doses of vitamin D supplementation during pregnancy on biochemical markers (serum 25(OH)D, calcium, phosphorus and alkaline phosphatase) in women and neonates, and on pregnancy and birth outcomes (gestational diabetes, pre-eclampsia, low birth weight, preterm births and stillbirths).ResultsPregnant women (N=350) in their first trimester were recruited and randomised to three treatment groups of vitamin D supplementation: 4000 IU/day (group A, n=120), 2000 IU/day (group B, n=115) or 400 IU/day (group C, n=115). Women and their newborn in group A had the lowest vitamin D deficiency at endline (endline: 75.9%; neonatal: 64.9%), followed by group B (endline: 84.9%; neonatal: 73.7%) and then the control group (endline: 90.2%; neonatal: 91.8%). Vitamin D deficiency was significantly lower in group A than in group C (p=0.006) among women at endline and lower in both groups A and B than in the control group (p=0.001) in neonates. Within groups, serum 25(OH)D was significantly higher between baseline and endline in group A and between maternal baseline and neonatal levels in groups A and B. Participant serum 25(OH)D levels at the end of the trial were positively correlated with those in intervention group A (4000 IU/day) (β=4.16, 95% CI 1.6 to 6.7, p=0.002), with food group consumption (β=0.95, 95% CI 0.01 to 1.89, p=0.047) and with baseline levels of serum 25(OH)D (β=0.43, 95% CI 0.29 to 0.58, p<0.0001).ConclusionThe evidence provided in our study indicates that vitamin D supplementation of 4000 IU/day was more effective in reducing vitamin D deficiency among pregnant women and in improving serum 25(OH)D levels in mothers and their neonates compared with 2000 IU/day and 400 IU/day.Trial registration number NCT02215213.