Factors influencing RNA yield from placenta tissue.

High yield and integrity of placental RNA are crucial for placental transcriptomics studies. We assessed the effects of time to placental collection post-delivery; tissue storage, amount and method used for extraction; mode of delivery; and tissue type on total RNA yield. The optimal protocol for RNA extraction from placental tissue includes cryofreezing of the sample upon collection and RNA extraction from 50 mg of tissue using TRIzol reagent. Decidua yielded highest RNA quantity/mg of tissue, followed by villous tissue and the chorion. Comparisons with murine kidney and HEK293T show lower placental RNA yield, likely due to highly dense and heterogeneous tissue make-up and potential high placental nuclease activity.

Elevated Maternal Folate Status and Changes in Maternal Prolactin, Placental Lactogen and Placental Growth Hormone Following Folic Acid Food Fortification: Evidence from Two Prospective Pregnancy Cohorts.

Folic acid (FA) food fortification in Australia has resulted in a higher-than-expected intake of FA during pregnancy. High FA intake is associated with increased insulin resistance and gestational diabetes. We aimed to establish whether maternal one-carbon metabolism and hormones that regulate glucose homeostasis change in healthy pregnancies post-FA food fortification. Circulating folate, B12, homocysteine, prolactin (PRL), human placental lactogen (hPL) and placental growth hormone (GH2) were measured in early pregnancy maternal blood in women with uncomplicated pregnancies prior to (SCOPE: N = 604) and post (STOP: N = 711)-FA food fortification. FA food fortification resulted in 63% higher maternal folate. STOP women had lower hPL (33%) and GH2 (43%) after 10 weeks of gestation, but they had higher PRL (29%) and hPL (28%) after 16 weeks. FA supplementation during pregnancy increased maternal folate and reduced homocysteine but only in the SCOPE group, and it was associated with 54% higher PRL in SCOPE but 28% lower PRL in STOP. FA food fortification increased maternal folate status, but supplements no longer had an effect, thereby calling into question their utility. An altered secretion of hormones that regulate glucose homeostasis in pregnancy could place women post-fortification at an increased risk of insulin resistance and gestational diabetes, particularly for older women and those with obesity.

Sexual Assault Labels, Compassion for Others, Self-Compassion, and Victim Blaming.

Two studies examined whether label preferences (survivor, victim, neither/other/both) and past assault experience (assaulted or not assaulted) are related to compassion for others, self-compassion, rape myth acceptance, and cognitive distortions surrounding rape. Findings indicate that a preference for a victim label is related to more negative outcomes (e.g., propensity to victim-blame, less compassion for others) compared to those advocating for a survivor label or falling into a neither/other/both category. Furthermore, those who experienced sexual assault have significantly lower self-compassion compared to those who do not experience sexual assault. Implications for the impact of labels are discussed.

High Folate, Perturbed One-Carbon Metabolism and Gestational Diabetes Mellitus.

Folate is a dietary micronutrient essential to one-carbon metabolism. The World Health Organisation recommends folic acid (FA) supplementation pre-conception and in early pregnancy to reduce the risk of fetal neural tube defects (NTDs). Subsequently, many countries (~92) have mandatory FA fortification policies, as well as recommendations for periconceptional FA supplementation. Mandatory fortification initiatives have been largely successful in reducing the incidence of NTDs. However, humans have limited capacity to incorporate FA into the one-carbon metabolic pathway, resulting in the increasingly ubiquitous presence of circulating unmetabolised folic acid (uFA). Excess FA intake has emerged as a risk factor in gestational diabetes mellitus (GDM). Several other one-carbon metabolism components (vitamin B12, homocysteine and choline-derived betaine) are also closely entwined with GDM risk, suggesting a role for one-carbon metabolism in GDM pathogenesis. There is growing evidence from in vitro and animal studies suggesting a role for excess FA in dysregulation of one-carbon metabolism. Specifically, high levels of FA reduce methylenetetrahydrofolate reductase (MTHFR) activity, dysregulate the balance of thymidylate synthase (TS) and methionine synthase (MTR) activity, and elevate homocysteine. High homocysteine is associated with increased oxidative stress and trophoblast apoptosis and reduced human chorionic gonadotrophin (hCG) secretion and pancreatic ß-cell function. While the relationship between high FA, perturbed one-carbon metabolism and GDM pathogenesis is not yet fully understood, here we summarise the current state of knowledge. Given rising rates of GDM, now estimated to be 14% globally, and widespread FA food fortification, further research is urgently needed to elucidate the mechanisms which underpin GDM pathogenesis.

Using family planning service statistics to inform model-based estimates of modern contraceptive prevalence.

The annual assessment of Family Planning (FP) indicators, such as the modern contraceptive prevalence rate (mCPR), is a key component of monitoring and evaluating goals of global FP programs and initiatives. To that end, the Family Planning Estimation Model (FPEM) was developed with the aim of producing survey-informed estimates and projections of mCPR and other key FP indictors over time. With large-scale surveys being carried out on average every 3-5 years, data gaps since the most recent survey often exceed one year. As a result, survey-based estimates for the current year from FPEM are often based on projections that carry a larger uncertainty than data informed estimates. In order to bridge recent data gaps we consider the use of a measure, termed Estimated Modern Use (EMU), which has been derived from routinely collected family planning service statistics. However, EMU data come with known limitations, namely measurement errors which result in biases and additional variation with respect to survey-based estimates of mCPR. Here we present a data model for the incorporation of EMU data into FPEM, which accounts for these limitations. Based on known biases, we assume that only changes in EMU can inform FPEM estimates, while also taking inherent variation into account. The addition of this EMU data model to FPEM allows us to provide a secondary data source for informing and reducing uncertainty in current estimates of mCPR. We present model validations using a survey-only model as a baseline comparison and we illustrate the impact of including the EMU data model in FPEM. Results show that the inclusion of EMU data can change point-estimates of mCPR by up to 6.7 percentage points compared to using surveys only. Observed reductions in uncertainty were modest, with the width of uncertainty intervals being reduced by up to 2.7 percentage points.

Sleep-related monitoring on awakening mediates the relationship between insomnia-related interpretive bias and insomnia symptoms using the insomnia ambiguity paradigm.

A number of studies have examined and confirmed the presence of a sleep-related interpretive bias amongst poor sleepers and individuals with insomnia using an insomnia ambiguity task. This study explored possible mechanisms underlying the relationship between interpretive bias and insomnia using the insomnia ambiguity task. More importantly, the possible mediating role of sleep-associated monitoring, sleep preoccupation, sleep anticipatory anxiety and generalized anxiety was also examined. A total of N = 176 participants were stratified into normal sleepers and those displaying insomnia symptoms. Participants completed an online version of the insomnia ambiguity task and questionnaire measures pertaining to sleep and anxiety. Data concerning task response time and time of testing were also collected. Individuals in the insomnia symptom group presented significantly higher sleep-related interpretive bias scores compared to normal sleepers. When sleepiness, sleep-associated monitoring, sleep preoccupation, sleep anticipatory anxiety and generalized anxiety were controlled for, only monitoring on awakening predicted sleep-related interpretive bias. Multiple mediation modelling demonstrated that sleep-associated monitoring on awakening mediated the relationship between interpretive bias and insomnia symptoms. The current outcomes are consistent with previous research, supporting the notion that insomnia is characterized by a disorder-consistent interpretive bias. Furthermore, monitoring for insomnia-consistent cues on awakening appears to mediate group differences in interpretive bias.

Microbiota composition in bilateral healthy breast tissue and breast tumors.

PURPOSE: Previous reports suggest that a complex microbiome exists within the female human breast that might contribute to breast cancer etiology. The purpose of this pilot study was to assess the variation in microbiota composition by breast side (left versus right) within individual women and compare the microbiota of normal and breast tumor tissue between women. We aimed to determine whether microbiota composition differs between these groups and whether certain bacterial taxa may be associated with breast tumors. METHODS: Bilateral normal breast tissue samples (n = 36) were collected from ten women who received routine mammoplasty procedures. Archived breast tumor samples (n = 10) were obtained from a biorepository. DNA was extracted, amplified, and sequenced. Microbiota data were analyzed using QIIME and RStudio. RESULTS: The most abundant phyla in both tumor and normal tissues were Bacteroidetes, Firmicutes, Proteobacteria, and Actinobacteria. There were statistically significant differences in the relative abundance of various bacterial taxa between groups. Alpha diversity (Simpson's index) was significantly higher in normal compared to tumor samples (0.968 vs. 0.957, p = 0.022). Based on unweighted UniFrac measures, breast tumor samples clustered distinctly from normal samples (R2 = 0.130; p = 0.01). Microbiota composition in normal samples clustered within women (R2 = 0.394; p = 0.01) and by breast side (left or right) within a woman (R2 = 0.189; p = 0.03). CONCLUSION: Significant differences in diversity between tumor and normal tissue and in composition between women and between breasts of the same woman were identified. These results warrant further research to investigate the relationship between microbiota and breast cancer.

Family Planning During and After the West African Ebola Crisis.

The West African Ebola outbreak of 2013-2016 had the potential to devastate family planning programs in affected countries, which had made great progress in years prior. We examine monthly provision of family planning service statistics from government sources for Liberia and Sierra Leone from 6 months before the first Ebola case to 24 months after the last Ebola case to measure the impact during and after the epidemic. By calculating the couple-years of protection from service statistics, we find that family planning distribution declined by 65 percent in Liberia and 23 percent in Sierra Leone at the peak of the epidemic. Two years after Ebola, Liberia's average monthly contraception distribution is 39 percent above precrisis levels, while distribution in Sierra Leone increased by 27 percent, findings echoed in data from the Demographic and Health Survey and Multiple Indicator Cluster Survey. Increased contraceptive use comes from implants in both countries, and injectables in Liberia. This study indicates that the family planning sector can recover, and continue to improve, following a significant disruption and is a lesson in resilience.

Effects of a Self-Compassion Break Induction on Self-Reported Stress, Self-Compassion, and Depressed Mood.

INTRODUCTION: The objective of the study was to determine whether practicing a self-compassion induction would reduce self-reported stress, depressed mood, and increase self-compassion in a randomized controlled study measuring variables of interest at two time points spanning three weeks. METHOD: Participants were 129 students (91 females and 38 males, Mage = 19.47, SD = 3.20) divided into three groups: Self-compassion Break, Time Management Control Group, and a No-Induction Control Group. Participants were part of the general student body and were not diagnosed or screened for clinical depression. Self-compassion, stress, and depressed mood were assessed at baseline and follow-up. Participants practiced their respective inductions over three weeks after being guided through their respective inductions at Time 1. RESULTS: There were no significant differences between groups in outcome variables. Results of this study may indicate the importance of weekly group check-in meetings to facilitate positive change as findings in this study did not mirror those of similar studies using frequent meetings.

Setting the stage for strengthened annual monitoring of family planning program performance at the state/national level in Myanmar.

Background: Although Myanmar has made good progress in family planning by increased contraceptive prevalence rate (CPR) from 41% in 2007 to 52.2% in 2016, it remains lower than the target of 60% by 2020. There are also huge disparities sub-nationally, ranging from 25% to 60%. While there is a strong need to monitor the progress of family planning program regularly at the national and sub-national level, Myanmar has limited surveys, data quality and methodological issues in its Health Management Information System (HMIS), and a scattered rollout of the Logistic Management Information System (LMIS). Methods: To identify viable options for annual monitoring, four data sources: modelled contraceptive prevalence rate for modern methods (mCPR) estimates from Track20's  Family Planning Estimation Tool (FPET); method-specific prevalence from the  2015-16 Myanmar Demographic and Health Survey (DHS); mCPR estimates and method prevalence from  HMIS and estimates of modern method use (EMU) based on commodity consumption data from LMIS, were compared for the years 2015-2017. Estimates of mCPR from HMIS were tested for accuracy based on whether they fell within the 95% confidence interval of mCPR estimates from the FPET for the corresponding years. EMU from LMIS was also tested for those years and states/regions where available. Results: For annual tracking of mCPR, direct estimates of HMIS were considered carefully, as they were much higher than those of the DHS survey and were not matched by FPET results, except in Chin and Kayin. To monitor the method mix, HMIS data can be used as these are similar pattern with DHS in both national and State/Regional level except Chin and Kayin. LMIS could be used in annual tracking when there are high reporting rates and valid information of consumption. Conclusions: Track20's FPET is the method of choice to get valid information for annual monitoring of family planning program.

Using Evidence to Drive Impact: Developing the FP Goals Impact Matrix.

When designing a family planning (FP) strategy, decision-makers can choose from a wide range of interventions designed to expand access to and develop demand for FP. However, not all interventions will have the same impact on increasing modern contraceptive prevalence (mCP). Understanding the existing evidence is critical to planning successful and cost-effective programs. The Impact Matrix is the first comprehensive summary of the impact of a full range of FP interventions on increasing mCP using a single comparable metric. It was developed through an extensive literature review with input from the wider FP community, and includes 138 impact factors highlighting the range of effectiveness observed across categories and subcategories of FP interventions. The Impact Matrix is central to the FP Goals model, used to project scenarios of mCP growth that help decision-makers set realistic goals and prioritize investments. Development of the Impact Matrix, evidence gaps identified, and the contribution to FP Goals are discussed.

Reconsidering Forced Labels: Outcomes of Sexual Assault Survivors Versus Victims (and Those Who Choose Neither).

The goal of the current study was to assess the effects of self-labeling on attitudes related to victim- and self-blaming and self-compassion outcomes in 85 participants (75 women, 10 men) who have experienced sexual assault. Participants classified themselves as either a survivor, victim, or neither survivor nor victim of sexual assault. Regardless of self-classification, groups did not differ in victim-blaming (rape myth acceptance), self-blaming, or levels of self-compassion. Implications for language and forced labels of those who have experienced sexual assault are discussed.

Can Family Planning Service Statistics Be Used to Track Population-Level Outcomes?

The need for annual family planning program tracking data under the Family Planning 2020 (FP2020) initiative has contributed to renewed interest in family planning service statistics as a potential data source for annual estimates of the modern contraceptive prevalence rate (mCPR). We sought to assess (1) how well a set of commonly recorded data elements in routine service statistics systems could, with some fairly simple adjustments, track key population-level outcome indicators, and (2) whether some data elements performed better than others. We used data from 22 countries in Africa and Asia to analyze 3 data elements collected from service statistics: (1) number of contraceptive commodities distributed to clients, (2) number of family planning service visits, and (3) number of current contraceptive users. Data quality was assessed via analysis of mean square errors, using the United Nations Population Division World Contraceptive Use annual mCPR estimates as the "gold standard." We also examined the magnitude of several components of measurement error: (1) variance, (2) level bias, and (3) slope (or trend) bias. Our results indicate modest levels of tracking error for data on commodities to clients (7%) and service visits (10%), and somewhat higher error rates for data on current users (19%). Variance and slope bias were relatively small for all data elements. Level bias was by far the largest contributor to tracking error. Paired comparisons of data elements in countries that collected at least 2 of the 3 data elements indicated a modest advantage of data on commodities to clients. None of the data elements considered was sufficiently accurate to be used to produce reliable stand-alone annual estimates of mCPR. However, the relatively low levels of variance and slope bias indicate that trends calculated from these 3 data elements can be productively used in conjunction with the Family Planning Estimation Tool (FPET) currently used to produce annual mCPR tracking estimates for FP2020.

Modern contraceptive use, unmet need, and demand satisfied among women of reproductive age who are married or in a union in the focus countries of the Family Planning 2020 initiative: a systematic analysis using the Family Planning Estimation Tool.

BACKGROUND: The London Summit on Family Planning in 2012 inspired the Family Planning 2020 (FP2020) initiative and the 120×20 goal of having an additional 120 million women and adolescent girls become users of modern contraceptives in 69 of the world's poorest countries by the year 2020. Working towards achieving 120 × 20 is crucial for ultimately achieving the Sustainable Development Goals of universal access and satisfying demand for reproductive health. Thus, a performance assessment is required to determine countries' progress. METHODS: An updated version of the Family Planning Estimation Tool (FPET) was used to construct estimates and projections of the modern contraceptive prevalence rate (mCPR), unmet need for, and demand satisfied with modern methods of contraception among women of reproductive age who are married or in a union in the focus countries of the FP2020 initiative. We assessed current levels of family planning indicators and changes between 2012 and 2017. A counterfactual analysis was used to assess if recent levels of mCPR exceeded pre-FP2020 expectations. FINDINGS: In 2017, the mCPR among women of reproductive age who are married or in a union in the FP2020 focus countries was 45·7% (95% uncertainty interval [UI] 42·4-49·1), unmet need for modern methods was 21·6% (19·7-23·9), and the demand satisfied with modern methods was 67·9% (64·4-71·1). Between 2012 and 2017 the number of women of reproductive age who are married or in a union who use modern methods increased by 28·8 million (95% UI 5·8-52·5). At the regional level, Asia has seen the mCPR among women of reproductive age who are married or in a union grow from 51·0% (95% UI 48·5-53·4) to 51·8% (47·3-56·5) between 2012 and 2017, which is slow growth, particularly when compared with a change from 23·9% (22·9-25·0) to 28·5% (26·8-30·2) across Africa. At the country level, based on a counterfactual analysis, we found that 61% of the countries that have made a commitment to FP2020 exceeded pre-FP2020 expectations for modern contraceptive use. Country success stories include rapid increases in Kenya, Mozambique, Malawi, Lesotho, Sierra Leone, Liberia, and Chad relative to what was expected in 2012. INTERPRETATION: Whereas the estimate of additional users up to 2017 for women of reproductive age who are married or in a union would suggest that the 120 × 20 goal for all women is overly ambitious, the aggregate outcomes mask the diversity in progress at the country level. We identified countries with accelerated progress, that provide inspiration and guidance on how to increase the use of family planning and inform future efforts, especially in countries where progress has been poor. FUNDING: The Bill & Melinda Gates Foundation, through grant support to the University of Massachusetts Amherst and Avenir Health.

Only a small proportion of patients with first episode psychosis come via prodromal services: a retrospective survey of a large UK mental health programme.

BACKGROUND: Little is known about patients with a first episode of psychosis (FEP) who had first presented to prodromal services with an "at risk mental state" (ARMS) before making the transition to psychosis. We set out to identify the proportion of patients with a FEP who had first presented to prodromal services in the ARMS state, and to compare these FEP patients with FEP patients who did not have prior contact with prodromal services. METHODS: In this study information on 338 patients aged ≤37 years who presented to mental health services between 2010 and 2012 with a FEP was examined. The data on pathways to care, clinical and socio-demographic characteristics were extracted from the Biomedical Research Council Case Register for the South London and Maudsley NHS Trust. RESULTS: Over 2 years, 14 (4.1% of n = 338) young adults presented with FEP and had been seen previously by the prodromal services. These ARMS patients were more likely to enter their pathway to psychiatric care via referral from General Practice, be born in the UK and to have had an insidious mode of illness onset than FEP patients without prior contact with the prodromal services. CONCLUSIONS: In the current pathways to care configuration, prodromal services are likely to prevent only a few at-risk individuals from transitioning to psychosis even if effective preventative treatments become available.

Structure and multistate function of the transmembrane electron transporter CcdA.

The mechanism by which transmembrane reductases use a single pair of cysteine residues to relay electrons between protein substrates across biological membranes is a long-standing mystery in thiol-redox biochemistry. Here we show the NMR structure of a reduced-state mimic of archaeal CcdA, a protein that transfers electrons across the inner membrane, by using a redox-active NMR sample. The two cysteine positions in CcdA are separated by 20 Å. Whereas one is accessible to the cytoplasm, the other resides in the protein core, thus implying that conformational exchange is required for periplasmic accessibility. In vivo mixed disulfide-trapping experiments validated the functional positioning of the cysteines, and in vitro accessibility results confirmed conformational exchange. Our NMR and functional data together show the existence of multiple conformational states and suggest a four-state model for relaying electrons from cytosolic to periplasmic redox substrates.

Helix stabilization precedes aqueous and bilayer-catalyzed fiber formation in islet amyloid polypeptide.

Islet amyloid polypeptide (IAPP) is an unstructured polypeptide hormone that is cosecreted with insulin. In patients with type 2 diabetes, IAPP undergoes a transition from its natively disordered state to a highly ordered, all-beta-strand amyloid fiber. Although predominantly disordered, IAPP transiently samples alpha-helical structure in solution. IAPP adopts a fully helical structure when bound to membrane surfaces in a process associated with catalysis of amyloid formation. Here, we use spectroscopic techniques to study the structure of full-length, monomeric IAPP under amyloidogenic conditions. We observe that the residues with helical propensity in solution (1-22) also form the membrane-associated helix. Additionally, reduction of the N-terminal disulfide bond (Cys2-Cys7) decreases the extent of helix formed throughout this region. Through manipulation of sample conditions to increase or decrease the amount of helix, we show that the degree of helix formed affects the rate of amyloid assembly. Formation of helical structure is directly correlated with enhanced amyloid formation both on the membrane surface and in solution. These observations support suggested mechanisms in which parallel helix associations bring together regions of the peptide that could nucleate beta-strand structure. Remarkably, stabilization of non-amyloid structure appears to be a key intermediate in assembly of IAPP amyloid.

Interaction of membrane-bound islet amyloid polypeptide with soluble and crystalline insulin.

Islet amyloid polypeptide (IAPP, also known as amylin) is the major protein component of pancreatic amyloid fibers in type II diabetes and is normally cosecreted with insulin from the beta-cells of the pancreas. IAPP forms amyloid fibrils rapidly at concentrations well below those found in vivo, yet progression of type II diabetes occurs over many years. Insulin, a known inhibitor of IAPP fibrillogenesis, exists as a dense crystalline or near-crystalline core in the secretory vesicle, while IAPP localizes to the region between the crystal and the secretory vesicle membrane. In vitro, IAPP fibrillogenesis is both accelerated by lipid membranes and inhibited by monomeric insulin. In this work, we investigate insulin-IAPP-lipid interactions in vitro under conditions chosen to approximate native secretory vesicle physiology and the amyloid disease state. The effect of insulin on IAPP fibrillogenesis is investigated using fluorescence spectrometry. Additionally, interactions of IAPP and lipids with crystalline insulin are studied using fluorescence microscopy. We find that, while soluble states of insulin and IAPP do not interact significantly, large assemblies of either insulin (crystals) or IAPP (fibers) can lead to stable IAPP-insulin interactions. The results raise the possibility of multiple physiological interactions between these two beta-cell hormones.

Direct detection of transient alpha-helical states in islet amyloid polypeptide.

The protein islet amyloid polypeptide (IAPP) is a glucose metabolism associated hormone cosecreted with insulin by the beta-cells of the pancreas. In humans with type 2 diabetes, IAPP deposits as amyloid fibers. The assembly intermediates of this process are associated with beta-cell death. Here, we examine the rat IAPP sequence variant under physiological solution conditions. Rat IAPP is mechanistically informative for fibrillogenesis, as it samples intermediate-like states but does not progress to form amyloid. A central challenge was the development of a bacterial expression system to generate isotopically labeled IAPP without terminal tags, but which does include a eukaryotic post-translational modification. While optical spectroscopy shows IAPP to be natively unfolded, NMR chemical shifts of backbone and beta-carbon resonances reveal the sampling of alpha-helical states across a continuous stretch comprising approximately 40% of the protein. In addition, the manifestation of nonrandom coil chemical shifts is confirmed by the relative insensitivity of the amide proton chemical shifts to alterations in temperature. Intriguingly, the residues displaying helical propensity are conserved with the human sequence, suggesting a functional role for this conformational bias. The inability of rat IAPP to self assemble can be ascribed, in part, to several slowly exchanging conformations evident as multiple chemical shift assignments in the immediate vicinity of three proline residues residing outside of this helical region.