A Practical Guide to Rodent Islet Isolation and Assessment

Pancreatic islets of Langerhans secrete hormones that are vital to the regulation of blood glucose and are, therefore, a key focus of diabetes research. Purifying viable and functional islets from the pancreas for study is an intricate process. This review highlights the key elements involved with mouse and rat islet isolation, including choices of collagenase, the collagenase digestion process, purification of islets using a density gradient, and islet culture conditions. In addition, this paper reviews commonly used techniques for assessing islet viability and function, including visual assessment, fluorescent markers of cell death, glucose-stimulated insulin secretion, and intracellular calcium measurements. A detailed protocol is also included that describes a common method for rodent islet isolation that our laboratory uses to obtain viable and functional mouse islets for in vitro study of islet function, beta-cell physiology, and in vivo rodent islet transplantation. The purpose of this review is to serve as a resource and foundation for successfully procuring and purifying high-quality islets for research purposes

Treatment of Infections in Young Infants in Low- and Middle-Income Countries:A Systematic Review and Meta-analysis of Frontline Health Worker Diagnosis and Antibiotic Access

BACKGROUND: Inadequate illness recognition and access to antibiotics contribute to high case fatality from infections in young infants (<2 months) in low- and middle-income countries (LMICs). We aimed to address three questions regarding access to treatment for young infant infections in LMICs: (1) Can frontline health workers accurately diagnose possible bacterial infection (pBI)?; (2) How available and affordable are antibiotics?; (3) How often are antibiotics procured without a prescription? METHODS AND FINDINGS: We searched PubMed, Embase, WHO/Health Action International (HAI), databases, service provision assessments (SPAs), Demographic and Health Surveys, Multiple Indicator Cluster Surveys, and grey literature with no date restriction until May 2014. Data were identified from 37 published studies, 46 HAI national surveys, and eight SPAs. For study question 1, meta-analysis showed that clinical sign-based algorithms predicted bacterial infection in young infants with high sensitivity (87%, 95% CI 82%-91%) and lower specificity (62%, 95% CI 48%-75%) (six studies, n = 14,254). Frontline health workers diagnosed pBI in young infants with an average sensitivity of 82% (95% CI 76%-88%) and specificity of 69% (95% CI 54%-83%) (eight studies, n = 11,857) compared to physicians. For question 2, first-line injectable agents (ampicillin, gentamicin, and penicillin) had low variable availability in first-level health facilities in Africa and South Asia. Oral amoxicillin and cotrimoxazole were widely available at low cost in most regions. For question 3, no studies on young infants were identified, however 25% of pediatric antibiotic purchases in LMICs were obtained without a prescription (11 studies, 95% CI 18%-34%), with lower rates among infants <1 year. Study limitations included potential selection bias and lack of neonatal-specific data. CONCLUSIONS: Trained frontline health workers may screen for pBI in young infants with relatively high sensitivity and lower specificity. Availability of first-line injectable antibiotics appears low in many health facilities in Africa and Asia. Improved data and advocacy are needed to increase the availability and appropriate utilization of antibiotics for young infant infections in LMICs. REVIEW REGISTRATION: PROSPERO International prospective register of systematic reviews (CRD42013004586). Please see later in the article for the Editors' Summary

Short EpiData course: do participants use the data entry tool two years post-training?

SETTING: Training courses on data entry are few and far between compared to analysis. To address this gap, we conducted a short 2-day course on quality-assured data capture using EpiData for public health professionals in Bangalore and Puducherry, India, in 2013. OBJECTIVE: To determine the proportion of participants who used EpiData and taught the software to others during the 2-years after training and explore the reasons for doing/not doing so. DESIGN: Cross-sectional study with self-administered, semi-structured questionnaire developed using web-based Google Forms. We performed a manual thematic analysis to identify the major reasons for using/not using EpiData. RESULTS: Of 46 participants, 38 (83%) responded. Of 31 participants involved in research, 17 (55%) had used EpiData, of whom 6 (35%) had performed double entry and validation. Of the 14 who did not use EpiData, 11 had used MS Excel or SPSS/Epi Info for data entry. Of the 38 respondents, 29 (76%) had taught EpiData to other colleagues and students. Reasons for using EpiData included its user-friendliness, its being open access and the ease in preventing data entry errors. Reasons for not performing double entry included lack of time and manpower. CONCLUSION: The short course on EpiData was effective in knowledge transfer and provides a scalable model for incorporation into the teaching curricula of medical schools and research institutions