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To determine whether the mechanotransduction and pharmacomanipulation of A-kinase anchoring protein 13 (AKAP13) altered Hippo signaling pathway transcription and growth factors in granulosa cells. Primary ovarian insufficiency is the depletion or dysfunction of primordial ovarian follicles. In vitro activation of ovarian tissue in patients with primary ovarian insufficiency alters the Hippo and phosphatase and tensin homolog/phosphatidylinositol 3-kinase/protein kinase B/forkhead box O3 pathways. A-kinase anchoring protein 13 is found in granulosa cells and may regulate the Hippo pathway via F-actin polymerization resulting in altered nuclear yes-associated protein (YAP)/transcriptional coactivator with PDZ-binding motif coactivators and Tea domain family (TEAD) transcription factors.Laboratory studies.Translational science laboratory.None.COV434 cells, derived from a primary human granulosa tumor cell line, were studied under different cell density and well stiffness conditions. Cells were transfected with a TEAD-luciferase (TEAD-luc) reporter as well as expression constructs for AKAP13 or AKAP13 mutants and then treated with AKAP13 activators, inhibitors, and follicle-stimulating hormone.TEAD gene activation or inhibition was measured by TEAD-luciferase assays. The messenger ribonucleic acid levels of Hippo pathway signaling molecules, including connective tissue growth factor (CTGF), baculoviral inhibitors of apoptosis repeat-containing 5, Ankyrin repeat domain-containing protein 1, YAP1, and TEAD1, were measured by quantitative real-time polymerase chain reaction. Protein expressions for AKAP13, CTGF, YAP1, and TEAD1 were measured using Western blot.Increased TEAD-luciferase activity and expression of markers for cellular growth were associated with decreased cell density, increased well stiffness, and AKAP13 activator (A02) treatment. Additionally, decreased TEAD-luc activity and expression of markers for cellular growth were associated with AKAP13 inhibitor (A13) treatment, including a reduced expression of the BIRC5 and ANKRD1 (YAP-responsive genes) transcript levels and CTGF protein levels. There were no changes in TEAD-luc with follicle-stimulating hormone treatment, supporting Hippo pathway involvement in the gonadotropin-independent portion of folliculogenesis.These findings suggest that AKAP13 mediates Hippo-regulated changes in granulosa cell growth via mechanotransduction and pharmacomanipulation. The AKAP13 regulation of the Hippo pathway may represent a potential target for regulation of follicle activation.

This meta-analysis was conducted to 1) assess the quantity and dose of perioperatively dispensed opioids for benign hysterectomy by procedure route and 2) identify the predictors of persistent opioid use after the procedure DATA SOURCES: PubMed, Web of Science and Embase were systematically searched up to 25 March 2022.Studies reporting data on opioid dispensing among patients undergoing benign hysterectomy were considered eligible. The primary outcome was the dosage of opioids dispensed perioperatively (from 30 preoperative days to 21 postoperative days). The secondary outcome was the predictors of persistent opioid use after benign hysterectomy (from 3 months to 3 years postoperatively). Total opioid dispensing was measured in morphine milligram equivalents (MME) units.The random-effects model was used to pool the mean differences or odds ratios (ORs) and the corresponding 95% confidence intervals (CIs).Eight studies presenting data on 377,569 women undergoing benign hysterectomy were included. Of these women, 83% (95% CI: 81, 84) were dispensed opioids during the perioperative period. The average amount of perioperatively dispensed opioids was 143.5 MME (95% CI: 40, 247). Women undergoing vaginal hysterectomy were dispensed a significantly lower amount of opioids than those undergoing laparoscopic or abdominal hysterectomies. The overall rate of persistent opioid use after benign hysterectomy was 5% (95% CI: 2, 8). Younger patient age (OR 1.38 [95% CI: 1.17, 1.63]), smoking history (OR 1.87 [95% CI: 1.67, 2.10]), alcohol use (OR 3.16 [95% CI: 2.34, 4.27]), back pain (OR 1.50 [95% CI: 1.10, 2.05]) and fibromyalgia (OR 1.60 [95% CI: 1.39, 1.83]) were significantly associated with a higher risk of persistent opioid use after benign hysterectomy. However, there was no significant impact of hysterectomy route and operative complexity on persistent opioid use postoperatively.Perioperative opioid dispensing is significantly dependent on the route of hysterectomy, with the lowest dispensed MME of opioids for vaginal hysterectomy and the highest for abdominal hysterectomy. Nevertheless, hysterectomy route did not significantly predict persistent opioid use postoperatively, whereas younger age, smoking, alcohol use, back pain and fibromyalgia were significantly associated with persistent opioid use.

Antibiotics have become a mainstay of healthcare in the past century due to their activity against pathogens. This manuscript reviews the impact of antibiotic use on the intestinal microbiota in the context of mass drug administration (MDA). The importance of the gut microbiota to human metabolism and physiology is now well established, and antibiotic exposure may impact host health via collateral effects on the microbiota and its functions. To gain further insight into how gut microbiota respond to antibiotic perturbation and the implications for public health, factors that influence the impact of antibiotic exposure on the microbiota, potential health outcomes of antibiotic-induced microbiota alterations, and strategies that have the potential to ameliorate these wider antibiotic-associated microbiota perturbations are also reviewed. Graphical Abstract

Next-generation sequencing (NGS) workflows applied to bronchoalveolar lavage (BAL) fluid specimens could enhance the detection of respiratory pathogens, although optimal approaches are not defined. This study evaluated the performance of the Respiratory Pathogen ID/AMR (RPIP) kit (Illumina, Inc.) with automated Explify bioinformatic analysis (IDbyDNA, Inc.), a targeted NGS workflow enriching specific pathogen sequences and antimicrobial resistance (AMR) markers, and a complementary untargeted metagenomic workflow with in-house bioinformatic analysis. Compared to a composite clinical standard consisting of provider-ordered microbiology testing, chart review, and orthogonal testing, both workflows demonstrated similar performances. The overall agreement for the RPIP targeted workflow was 65.6% (95% confidence interval, 59.2 to 71.5%), with a positive percent agreement (PPA) of 45.9% (36.8 to 55.2%) and a negative percent agreement (NPA) of 85.7% (78.1 to 91.5%). The overall accuracy for the metagenomic workflow was 67.1% (60.9 to 72.9%), with a PPA of 56.6% (47.3 to 65.5%) and an NPA of 77.2% (68.9 to 84.1%). The approaches revealed pathogens undetected by provider-ordered testing (Ureaplasma parvum, Tropheryma whipplei, severe acute respiratory syndrome coronavirus 2 [SARS-CoV-2], rhinovirus, and cytomegalovirus [CMV]), although not all pathogens detected by provider-ordered testing were identified by the NGS workflows. The RPIP targeted workflow required more time and reagents for library preparation but streamlined bioinformatic analysis, whereas the metagenomic assay was less demanding technically but required complex bioinformatic analysis. The results from both workflows were interpreted utilizing standardized criteria, which is necessary to avoid reporting nonpathogenic organisms. The RPIP targeted workflow identified AMR markers associated with phenotypic resistance in some bacteria but incorrectly identified

BACKGROUND Family history (FH) is a powerful tool in screening and testing for chronic disease, oncologic conditions, and other genetic diagnoses, and is dependent upon observing patterns of features inherited across generations. Despite acceptance of FH as a fundamental part of a genetic medicine evaluation, there are limited data in other medical fields about its utility and benefit to patients. To date, there are no published data pertaining to the clinical utility of FH to diagnose individuals with hypophosphatasia (HPP), a rare genetic low bone density condition. OBJECTIVE To compare FH recorded in the electronic medical record (EMR) of patients at risk of HPP before a genetic medicine encounter to that learned during that clinical evaluation. METHODS We evaluated two patient populations at-risk of a rare metabolic condition-- hypophosphatasia (HPP)—for the quality and quantity of FH data in their medical records. Population 1 was derived from patients seeking evaluation for a low bone density diagnosis at the Johns Hopkins Greenberg Center for Skeletal Dysplasias (GCSD); population 2 was identified through a targeted electronic medical record (EMR) query of low serum alkaline phosphatase (AP) measurements obtained through the Johns Hopkins Clinical Laboratory. RESULTS In population 1 (n = 38), 27 (71%) were confirmed to have HPP. Of these, 14 (52%) presented with FH information in the EMR from 3 or more family members, and in 4 (15%) FH was suggestive of HPP. In population 2 (n=348), a similar proportion of subjects had 3 or more family members mentioned in the EMR (183 or 53%), but only 3 subjects (1%) included sufficient detail to determine that the family history was suggestive of HPP. Notably, once all patients in population 1 completed a medical genetics evaluation and HPP was confirmed, 20 (74%) had probable affected family members identified through obtaining and analyzing a pedigree. After cascade testing was offered to these family members, 17 patients (71%) from population 1 had at least one family member with HPP confirmed at a molecular level. CONCLUSIONS Based on these results, we propose that a full genetic medicine evaluation of those subjects from population 2 at highest risk of HPP would confirm HPP in many of them and identify similarly affected family members. We propose that taking a FH refines the diagnostic precision for patients with low bone density and identifies affected family members, and that HPP is likely more prevalent than previously thought. CLINICALTRIAL n/a

Summary Background Child stunting reduces survival and impairs neurodevelopment. We tested the independent and combined effects of improved water, sanitation, and hygiene (WASH), and improved infant and young child feeding (IYCF) on stunting and anaemia in in Zimbabwe. Methods We did a cluster-randomised, community-based, 2 × 2 factorial trial in two rural districts in Zimbabwe. Clusters were defined as the catchment area of between one and four village health workers employed by the Zimbabwe Ministry of Health and Child Care. Women were eligible for inclusion if they permanently lived in clusters and were confirmed pregnant. Clusters were randomly assigned (1:1:1:1) to standard of care (52 clusters), IYCF (20 g of a small-quantity lipid-based nutrient supplement per day from age 6 to 18 months plus complementary feeding counselling; 53 clusters), WASH (construction of a ventilated improved pit latrine, provision of two handwashing stations, liquid soap, chlorine, and play space plus hygiene counselling; 53 clusters), or IYCF plus WASH (53 clusters). A constrained randomisation technique was used to achieve balance across the groups for 14 variables related to geography, demography, water access, and community-level sanitation coverage. Masking of participants and fieldworkers was not possible. The primary outcomes were infant length-for-age Z score and haemoglobin concentrations at 18 months of age among children born to mothers who were HIV negative during pregnancy. These outcomes were analysed in the intention-to-treat population. We estimated the effects of the interventions by comparing the two IYCF groups with the two non-IYCF groups and the two WASH groups with the two non-WASH groups, except for outcomes that had an important statistical interaction between the interventions. This trial is registered with ClinicalTrials.gov, number NCT01824940. Findings Between Nov 22, 2012, and March 27, 2015, 5280 pregnant women were enrolled from 211 clusters. 3686 children born to HIV-negative mothers were assessed at age 18 months (884 in the standard of care group from 52 clusters, 893 in the IYCF group from 53 clusters, 918 in the WASH group from 53 clusters, and 991 in the IYCF plus WASH group from 51 clusters). In the IYCF intervention groups, the mean length-for-age Z score was 0·16 (95% CI 0·08–0·23) higher and the mean haemoglobin concentration was 2·03 g/L (1·28–2·79) higher than those in the non-IYCF intervention groups. The IYCF intervention reduced the number of stunted children from 620 (35%) of 1792 to 514 (27%) of 1879, and the number of children with anaemia from 245 (13·9%) of 1759 to 193 (10·5%) of 1845. The WASH intervention had no effect on either primary outcome. Neither intervention reduced the prevalence of diarrhoea at 12 or 18 months. No trial-related serious adverse events, and only three trial-related adverse events, were reported. Interpretation Household-level elementary WASH interventions implemented in rural areas in low-income countries are unlikely to reduce stunting or anaemia and might not reduce diarrhoea. Implementation of these WASH interventions in combination with IYCF interventions is unlikely to reduce stunting or anaemia more than implementation of IYCF alone. Funding Bill & Melinda Gates Foundation, UK Department for International Development, Wellcome Trust, Swiss Development Cooperation, UNICEF, and US National Institutes of Health.

OBJECTIVES: To determine the seroprevalence of HIV and identify associated risk factors among inmates at the Belize Central Prison, managed by the Kolbe Foundation, Belize. METHODS: A voluntary sample of 623 participants was obtained from the male inmate population incarcerated during the period from 15 January to 5 March 2005. HIV serostatus was determined on location using the Abbott Determine Assay for HIV-1/2 for screening, and the MedMira MiraWell Rapid HIV-1/2 Test for confirmatory testing. Remaining serum was tested by ELISA at the Central Medical Laboratory, Belize. Demographic and risk behavior data were collected using an interviewer administered pre-tested questionnaire. A multivariate logistic regression was used to adjust for potential confounders and to identify independent associations with HIV seropositivity. RESULTS:Of the 623 inmates in the sample, 25 tested positive for HIV-1/2 antibody for a seroprevalence of 4.0% (95% Confidence Interval 2.7, 6.0). After adjustment for confounding, HIV serostatus was positively associated with male-to-male sexual activity outside prison, age, and district of residence before current incarceration. CONCLUSIONS: The seroprevalence in the Central Prison was almost twice that estimated for the adult population of Belize in 2004 (2.4%). However, the social variables of importance to inmates appeared to reflect the epidemic in the general population, with the exception that male-to-male sex outside prison is likely more important to the male inmate population in Belize. The findings suggest that HIV is likely contracted by most inmates before their incarceration, largely due to same-sex activity.OBJETIVOS: Determinar la seroprevalencia al VIH e identificar los factores de riesgo asociados en hombres internados en la Prisión Central de Belice, administrada por la Fundación Kolbe. MÉTODOS: La muestra estuvo compuesta por 623 voluntarios hombres que se encontraban encarcelados entre el 15 de enero y el 5 de marzo de 2005. El estatus serológico con respecto al VIH se determinó en la prisión mediante la prueba de tamizaje Abbot Determine Assay y se confirmó con la prueba MedMira MiraWell Rapid, ambas para anticuerpos contra el VIH 1 y 2. El suero restante se analizó por ELISA en el Laboratorio Médico Central de Belice. Se recabaron los datos demográficos y sobre las conductas de riesgo mediante una encuesta preevaluada aplicada por un entrevistador. Se identificaron las asociaciones independientes con la seropositividad al VIH mediante análisis de regresión logística multifactorial ajustado por posibles factores de confusión. RESULTADOS: De los 623 prisioneros de la muestra, 25 resultaron positivos a anticuerpos contra el VIH-1/2, para una seroprevalencia de 4,0% (IC95%: 2,7% a 6,0%). Después de ajustar por los factores de confusión, la seropositividad se asoció con la actividad sexual con otros hombres fuera de la prisión, la edad y el distrito de residencia antes de su encarcelamiento actual. CONCLUSIONES: La seroprevalencia en la Prisión Central casi duplicó el estimado para la población adulta de Belice en 2004 (2,4%). Sin embargo, las variables sociales de importancia en los prisioneros parecieron reflejar la epidemia en la población general, aunque la relación sexual con hombres fuera de la prisión pareció tener mayor importancia en la población masculina encarcelada en Belice. Estos resultados indican que la mayoría de los prisioneros habría contraído la infección por el VIH antes de su encarcelamiento, en gran parte debido a prácticas homosexuales.

OBJECTIVE The increasing burdens of obesity and diabetes are two of the most prominent threats to the health of populations of developed and developing countries alike. The Central America Diabetes Initiative (CAMDI) is the first study to examine the prevalence of diabetes in Central America. RESEARCH DESIGN AND METHODS The CAMDI survey was a cross-sectional survey based on a probabilistic sample of the noninstitutionalized population of five Central American populations conducted between 2003 and 2006. The total sample population was 10,822, of whom 7,234 (67%) underwent anthropometry measurement and a fasting blood glucose or 2-h oral glucose tolerance test. RESULTS The total prevalence of diabetes was 8.5%, but was higher in Belize (12.9%) and lower in Honduras (5.4%). Of the screened population, 18.6% had impaired glucose tolerance/impaired fasting glucose. CONCLUSIONS As this population ages, the prevalence of diabetes is likely to continue to rise in a dramatic and devastating manner. Preventive strategies must be quickly introduced. Apparent changes in access to cheap, energy-dense food, urbanization, and adoption of sedentary lifestyles in the countries of Central America have raised concerns about the rapid emergence of obesity and diabetes in the region. Diabetes and related chronic conditions in Central America have been largely neglected by epidemiologic and surveillance programs in recent decades, however, because other issues, such as under-nutrition, infectious diseases, and armed conflict, were regarded as much more pressing health threats. The Central America Diabetes Initiative (CAMDI) is the first population-based multinational study to examine the prevalence of diabetes and risk factors in Central America. We report here the main findings from the multinational analyses of this survey.

To understand the mechanisms underlying the differential targeting of T-cell responses during HIV-1 disease progression.We performed a cross-sectional analysis of HIV specific CD8 T-cell responses in peripheral blood mononuclear cells (PBMC) obtained from 21 subjects with well characterized acute or early infection and 88 subjects with chronic HIV-1 infection. We also performed a longitudinal analysis of T-cell responses in five early infected subjects one of whom was studied extensively over a 4-year-period.PBMC were stimulated with pools of peptides encompassing all of the HIV-1 proteins in an interferon-gamma ELISpot assay. A mean entropy score was calculated for each peptide in the HIV-1 genome.The early infected group preferentially targeted variable peptides with higher entropy while responses towards more conserved peptides with lower entropy predominated in the group with chronic infection. In five early infected subjects followed longitudinally, responses to variable proteins declined while those to conserved proteins increased over time. In the subject who was followed for 4 years, epitopes in Vif and Nef were targeted early and escape occurred in three of these four epitopes. During the chronic phase of his infection, the early responses waned with an associated increase in breadth of T-cell responses mainly to Gag and Pol epitopes.Taken together, these data demonstrate that HIV-specific CD8 T cells are directed preferentially to the variable peptides in early infection but diminish in frequency during chronic disease, in large part due to cytotoxic T lymphocyte escape.

Chronic malnutrition, termed stunting, is defined as suboptimal linear growth, affects one third of children in developing countries, and leads to increased mortality and poor developmental outcomes. The causes of childhood stunting are unknown, and strategies to improve growth and related outcomes in children have only had modest impacts. Recent studies have shown that the ecosystem of microbes in the human gut, termed the microbiota, can induce changes in weight. However, the specific changes in the gut microbiota that contribute to growth remain unknown, and no studies have investigated the gut microbiota as a determinant of chronic malnutrition. We performed secondary analyses of data from two well-characterized twin cohorts of children from Malawi and Bangladesh to identify bacterial genera associated with linear growth. In a case-control analysis, we used the graphical lasso to estimate covariance network models of gut microbial interactions from relative genus abundances and used network analysis methods to select genera associated with stunting severity. In longitudinal analyses, we determined associations between these selected microbes and linear growth using between-within twin regression models to adjust for confounding and introduce temporality. Reduced microbiota diversity and increased covariance network density were associated with stunting severity, while increased relative abundance of Acidaminococcus sp. was associated with future linear growth deficits. We show that length growth in children is associated with community-wide changes in the gut microbiota and with the abundance of the bacterial genus, Acidaminococcus. Larger cohorts are needed to confirm these findings and to clarify the mechanisms involved.