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Phosphorylated biomarkers are crucial for our understanding of drug mechanism of action and dose selection during clinical trials, particularly for drugs that target protein kinases, such as DNA-damage-response (DDR) inhibitors. However, tissue fixation conditions needed to preserve DDR-specific phospho-biomarkers have not been previously investigated. Using xenograft tissues and tightly controlled formalin fixation conditions, we assessed how preanalytical factors affect phosphorylated DDR biomarkers pRAD50(Ser635), ɣH2AX(Ser139), pKAP1(Ser824), and non-phosphorylated biomarkers cMYC and ATM. Cold ischemia times ranged from 15 min to 6 hr, and the fixation duration ranged from 24 hr to 4 weeks. Epitopes pRAD50 and pKAP1 appeared the most labile assessed with staining loss after just 15 min of cold ischemia time, while ATM was more robust showing consistent expression up to 1 hr of cold ischemia. Notably, ɣH2AX expression was lost with formalin fixation over 48 hr. The use of core needle biopsies where possible and novel fixation methods such as the 2-step temperature-controlled formalin approach may improve phosphorylated biomarker preservation; however, practical challenges may affect wider clinical application. The most essential tissue-processing step when downstream analysis includes DDR phosphorylated biomarkers is immediate tissue submersion in formalin, without delay, upon excision from the patient, followed by room temperature fixation for 24 hr.

Background: Behaviour change interventions influence behaviour through causal processes called “mechanisms of action” (MoAs). Reports of such interventions and their evaluations often use inconsistent or ambiguous terminology. This includes the reporting of MoAs, creating challenges for searching, evidence synthesis and theory development. An ontology can help address these challenges by serving as a classification system that labels and defines classes for MoAs and their relationships. Aim: To develop an ontology of MoAs of behaviour change interventions.Methods: To develop the MoA Ontology, we (1) defined the ontology’s scope; (2) identified, labelled and defined the ontology’s classes; (3) refined the ontology by annotating (i.e., coding) MoAs in intervention reports; (4) refined the ontology via stakeholder review of the ontology’s comprehensiveness and clarity; (5) tested whether researchers could reliably apply the ontology to annotate MoAs in intervention reports; (6) refined the relationships between classes; (7) reviewed the alignment of the MoA Ontology with relevant ontologies, (8) reviewed the ontology’s alignment with the Theories and Techniques Project; and (9) published the ontology and created a machine-readable version.Results: An MoA was defined as “a process that is causally active in the relationship between a behaviour change intervention scenario and its outcome behaviour”. We created an initial MoA Ontology with 261 classes through Steps 2-5. Inter-rater reliability for annotating study reports using these classes was α=0.68 (“acceptable”) for researchers familiar with the ontology and α=0.47 for researchers unfamiliar with it. As a result of additional revisions (Steps 6-8), 21 further classes were added to the ontology resulting in 282 classes organised into seven hierarchical levels.Conclusions and implications: The MoA Ontology extensively captures MoAs of behaviour change interventions. The ontology can serve as a controlled vocabulary for MoAs to consistently describe and synthesise evidence about MoAs across diverse sources.

Since the final decades of the nineteenth century geographers have maintained a commitment to employing language that is accessible and understood by as many as possible. This disciplinary goal has motivated the making and use of maps, globes and other images such as photographs. Whilst confirming this argument, this article shows that geographers have also enjoyed wordplay and that they have persistently had recourse to verbal invention in communicating geographical ideas to a diversifying demographic of audiences across a widening geographical and disciplinary spectrum over the period 1887 to 1947. In tracing the historical and geographical variations of the term ‘geosophy’, the article shows how this word circulated perceptions of time and space and an informed philosophy of geographical ideals and materials. The recurring refrain of geosophy elucidates understandings of verbal and visual invention and emulation. It concludes that the plurality of definitions of the portmanteau word geosophy are commensurate with the perception of the plurality of worlds and the relativity of worlds of knowledge.

Evidence suggests a consistent correlation between vision impairments, social isolation, and cognitive decline. The National Eye Institute reports that African Americans have an increased risk of developing certain vision impairments such as cataracts, glaucoma, and diabetic retinopathy. At the same time, older African Americans often receive care from family members and this family care may act as a buffer against social isolation and resulting cognitive decline. Using data from 737 African Americans that participated in waves 5, 6, and 7 of the National Health and Aging Trends Study (NHATS), we explored associations between vision impairment, social isolation, and cognitive functioning. Results showed that vision impairment at round 5 was related to increased social isolation, and higher social isolation at round 5 was related to decreased delayed word recall scores at the same wave. No significant longitudinal associations were found between these constructs. Findings suggest that concurrent associations exist between sensory impairments, social isolation, and cognitive functioning, but that these relationships are not robust across time. Despite support provided by unpaid family caregivers, African American older adults with vision impairment are at an increased risk for concurrent social and cognitive challenges. It may be that family support of those with sensory impairments helps so that these impairments aren’t related to social isolation or cognitive functioning across time. Researchers and clinicians could benefit older African Americans with sensory impairments by providing and encouraging support during early stages of vision loss.

Background: Propofol and dexmedetomidine are routinely used in intensive care units (ICUs). Both are lipophilic, potentially leading to accumulation in adipose tissue. Limited evidence exists on what body weight to utilize in patients with morbid obesity. Objective: The purpose of this research was to evaluate the safety and efficacy of weight-based sedation with propofol and dexmedetomidine in ICU patients with morbid obesity. Methods: Retrospective review of ICU patients admitted from January 2018 to January 2020 who were sedated for ≥48 hours was performed. The primary outcome was the percentage of time within target sedation during the first 48 hours, stratified by body mass index (BMI) 2. Additional outcomes included adverse events and the infusion rate to achieve target sedation. Data were evaluated using descriptive statistics. Results: A total of 80 patients were analyzed. Patients on propofol with a BMI 2 were in their target Richmond Agitation-Sedation Scale (RASS) 11.7% versus 16.1% with a BMI ≥40 kg/m2 ( P = .580). Patients with a BMI 2 on dexmedetomidine were in their target RASS 27.6% versus 10.7% with a BMI ≥40 kg/m2 ( P = .053). Conclusion and Relevance: Body mass index did not significantly alter propofol target sedation attainment. However, patients with morbid obesity on dexmedetomidine demonstrated significantly less time in target sedation. An actual body weight dosing strategy of these drugs did not achieve desired target sedation for those with a BMI ≥40 versus BMI 2. These findings support future research on the optimal sedation dosing strategy in this patient population.

Purpose: The primary objective of this systematic review was to determine if frenotomy for ankyloglossia improves breastfeeding or speech outcomes in infants and children ages birth to 12.Method: L...

An assessment of the visual output of the eugenicist geographer and landscape heritage activist Vaughan Cornish (1862-1948) constitutes a void in scholarly studies by geographers and histor...

Objective To investigate the association of maternal body mass index (BMI) and recurrent pregnancy loss (RPL). Design Systematic review and meta-analysis. Setting Not applicable. Patient(s) A total of 3,833 women with RPL and 4,083 women as controls. Intervention(s) Studies were identified through a search of PubMed, Embase, Scopus, and Cochrane. Main Outcome Measure(s) The primary outcome of interest was RPL using the mean differences in maternal BMI as the predictor variable. The results of the meta-analysis were reported as the mean difference with a 95% confidence interval. Result(s) In total, 892 studies were reviewed. Pooled data from 25 studies suggested that the maternal BMI of women with a history of recurrent pregnancy loss was significantly higher than the BMI of controls, mean difference 0.7 kg/m2 [95% confidence interval 0.2–1.3]. Conclusion(s) These findings supported an association between maternal BMI and RPL. Large prospective studies are needed to evaluate the influence of maternal BMI on pregnancy outcomes in women with RPL.

The Emergency Department Return Visit Quality Program was launched in Ontario, Canada, to promote a culture of quality. It mandates the province's largest-volume emergency departments (EDs) to audit charts of patients who had a return visit leading to hospital admission, including some of their 72-hour all-cause return visits with admission and all of their 7-day ones with sentinel diagnoses (ie, acute myocardial infarction, subarachnoid hemorrhage, and pediatric sepsis), and submit their findings to a governmental agency. This provides an opportunity to identify possible adverse events and quality issues, which hospitals can then address through quality improvement initiatives. A group of emergency physicians with quality improvement expertise analyzed the submitted audits and accompanying narrative templates, using a general inductive approach to develop a novel classification of recurrent quality themes. Since the Return Visit Quality Program launched in 2016, 125,698 return visits with admission have been identified, representing 0.93% of the 86 participating EDs' 13,559,664 visits. Overall, participating hospitals have conducted 12,852 detailed chart audits, uncovering 3,010 (23.4%) adverse events/quality issues and undertaking hundreds of quality improvement provincewide projects as a result. The inductive analysis revealed 11 recurrent themes, classified into 3 groupings: patient characteristics (ie, patient risk profile and elder care), ED team actions or processes (ie, physician cognitive lapses, documentation, handover/communication between providers, radiology, vital signs, and high-risk medications or medication interactions), and health care system issues (ie, discharge planning/community follow-up, left against medical advice/left without being seen, and imaging/testing availability). The Return Visit Quality Program is the largest mandatory audit program for EDs and provides a novel approach to identify local adverse events/quality issues to target for improved patient safety and quality of care. It provides a blueprint for health system leaders to enable clinicians to develop an approach to organizational quality, as well as for teams to construct an audit system that yields defined issues amenable to improvement.

Posttranslational modifications (PTMs) on histone tails spatiotemporally dictate mammalian neural stem cell (NSC) fate. Bone morphogenetic protein 4 (BMP4), a member of the transforming growth factor β (TGF-β) superfamily, suppresses NSC proliferation and fosters differentiation into astroglial cells. Whether PTMs mediate these effects of BMP4 is unknown. Here we demonstrate that BMP4 signaling causes a net reduction in cellular histone H3 lysine 4 trimethylation (H3K4me3), an active histone mark at promoters of genes associated with human NSC proliferation. We also show that H3K4me3 reduction by BMP4 is mediated by decreased expression of SETD1A and WDR82, two methyltransferase components of SETD1A-COMPASS. Down-regulation of these components decreases expression of key genes expressed in hNSCs, while ectopic expression via transfection dedifferentiates human astrocytes (HAs). These observations suggest that BMP4 influences NSC fate by regulating PTMs and altering chromatin structure.SIGNIFICANCE STATEMENTBMP4 is critical in determining hNSC fate. Whether histone posttranslational modifications (PTM) mediate the effects of BMP4 is unknown. Here we report that H3K4me3, brought about by its methyltransferases SETD1A and WDR82, at promoters of stem cell genes OCT4 and NESTIN is involved in human neural stem cell (hNSC) maintenance. BMP4 promotes hNSC astroglial differentiation in part through reduction of SETD1A and WDR82 and thus decreased frequency of H3K4me3 at the promoters of these genes. These results provide evidence that BMP4 promotes hNSC differentiation through a potential epigenetic mechanism and extend our understanding of the role of histone PTM in central nervous system development.

This paper offers an account of the emergence and development of the magic lantern practices of the Royal Geographical Society (RGS) in London. It engages with the incremental, and contested, introduction of the lantern and lantern slides into the meetings of the RGS. The paper first assesses approaches to the magic lantern adopted by historical geographers. It then explores the close connections between the uptake of the technology at the RGS and contemporary geographical societies in Britain and France, and in metropolitan scientific societies and entertainment organisations located close to the RGS. It elucidates the society's engagement with the lantern and the diverse perceptions of the medium within that institution. It traces the lantern's gradual introduction across three RGS meetings between 1886 and 1888, during the society's wider reformation and its promotion of geographical education and science. This recasts Halford Mackinder's ‘On the Scope and Methods of Geography’ as a lantern-slide lecture. Addressing how, when and why this medium became a part of the RGS's knowledge-making practices, and examining the debates around the use of this technology, the paper relates this process to a broader context of visual practices, the professionalization of academic geography and understandings of nineteenth-century geographical modernity.

A substantial amount of evidence suggests that androgen signaling through classical androgen receptors is critical for both normal and pathologic ovarian physiology. Specifically, we and others have shown that, in mouse granulosa cells, androgen actions through both extranuclear and nuclear androgen receptor signaling are critical for normal follicle development and ovulation. Here, we show that androgens through the PI3K/Akt pathway rapidly (within minutes) phosphorylate and inhibit activity of the Polycomb group protein enhancer of zeste homolog 2 (Ezh2). Over the course of 24 to 48 hours, androgens then induce expression of the microRNA miR-101, which targets Ezh2 messenger RNA (mRNA), leading to a nearly complete loss of Ezh2 protein expression. This long-term androgen-induced loss of Ezh2 actions ultimately results in sustained reduction of the H3K27me3-repressive mark in the promoter region of the Runt-related transcription factor-1 (Runx1) gene, a luteinizing hormone (LH)–induced transcription factor essential for ovulation, leading to increased Runx1 mRNA expression. Accordingly, blocking androgen-induced inhibition of Ezh2 in vivo adversely affects LH-induced Runx1 mRNA expression and subsequent ovulation. Importantly, although estrogen treatment of granulosa cells similarly causes rapid activation of the PI3K/Akt pathway and short-term phosphorylation of Ezh2, it does not induce miR-101 expression and thereby does not reduce overall Ezh2 expression, demonstrating the androgen specificity of long-term Ezh2 suppression. Thus, this study provides insight regarding how androgen-induced extranuclear kinase signaling and intranuclear transcription through Ezh2 modifications may influence the expression pattern of genes, ultimately affecting various downstream physiological processes.

Anti-Müllerian hormone (AMH) is a member of the transforming growth factor-β superfamily and plays a crucial role in testicular and ovarian functions. In clinical practice, AMH is used as a diagnostic and/or prognostic marker in women in association with ovulation induction and in various pathophysiological conditions. Despite widespread clinical use of AMH, our mechanistic understanding of AMH actions in regulating follicular development is limited. Using a mouse model, we in this study report that in vivo AMH treatment while stalls follicular development and inhibits ovulation, also prevents follicular atresia. We further show that these AMH actions are mediated through induction of two miRNAs, miR-181a and miR-181b, which regulate various aspects of FSH signaling and follicular growth, ultimately affecting downstream gene expression and folliculogenesis. We also report that in this mouse model AMH pre-treatment prior to superovulation improves oocyte yield. These studies, therefore, offer new mechanistic insight into AMH actions in folliculogenesis and point toward potential utilization of AMH as a therapeutic agent.

Obesity is considered detrimental to women's reproductive health. Although most of the attention has been focused on the effects of obesity on hypothalamic function, studies suggest a multifactorial impact. In fact, obesity is associated with reduced fecundity even in women with regular cycles, indicating that there may be local ovarian effects modulating fertility. Here we describe a novel mechanism for leptin actions directly in the ovary that may account for some of the negative effects of obesity on ovarian function. We find that normal cycling, obese, hyperleptinemic mice fed with a high-fat diet are subfertile and ovulate fewer oocytes compared with animals fed with a normal diet. Importantly, we show that leptin induces expression of the neuropeptide cocaine- and amphetamine-regulated transcript (CART) in the granulosa cells (GCs) of ovarian follicles both in vitro and in vivo. CART then negatively affects intracellular cAMP levels, MAPK signaling, and aromatase mRNA expression, which leads to lower estradiol synthesis in GCs and altered ovarian folliculogenesis. Finally, in human samples from patients undergoing in vitro fertilization, we show a significant positive correlation between patient body mass index, CART mRNA expression in GCs, and CART peptide levels in follicular fluid. These observations suggest that, under obese conditions, CART acts as a local mediator of leptin in the ovary to cause ovarian dysfunction and reduced fertility.

In 2014 we began the first systematic study of water quality, specifically fecal contamination of drinking water in the Khumbu Valley, Sagarmatha National Park (SNP, Mt. Everest region), Nepal. Our goal was to identify coliform bacteria and E. coli in drinking water and groundwater-fed springs to generate a data set that will function as a base for potable water supplies and further monitoring. Sampling occurred in May (pre-monsoon summer) and early November (post-monsoon early winter) 2014. Sample sites were selected based on proximity to villages and primary use as a drinking water source. Overall, the data presented a predictable correlation between fecal contamination and both elevation and increasing population/tourist traffic. Drinking water within the study area met current World Health Organization drinking water standards for the physical properties of temperature (2.8°C - 13°C), pH (5.27 - 7.24), conductivity (14.5 - 133 mS) and TDS (7.24 - 65.5 ppm). Samples from the more populated, lower altitude areas had higher levels of E. coli. Samples collected and analyzed in May (pre-monsoon summer) had a higher level of E. coli and coliform bacteria than samples collected in November (post-monsoon early winter) suggesting a seasonal dependence overlaid on the population signature. Surface water typically had higher E. coli values than groundwater-fed springs. Temperature, total dissolved solids and conductivity generally decreased with increasing elevation, whereas pH increased with increasing elevation. There appears to be significant presence of fecal contamination of water sources due to a combination of tourism, elevation and seasons.

Glioma stem cells (GSCs), with neural stem cell (NSC) like properties are key factors behind therapeutic resistance in pediatric high-grade gliomas (HGG), the second most formidable childhood cancers. Targeting GSCs is a promising pathway for enhancing therapeutic effect and improving patient outcomes. Histone post-translational modifications (PTMs) regulate gene expression patterns dictate stem cell fate and regulate drug resistance. Tri-methylation on histone 3 lysine 4 (H3K4me3) has been associated with human NSCs and GSCs, with its reduction results in cell differentiation and increases chemotherapeutic efficacy in pediatric HGGs. Human SETD1A facilitates H3K4me3 on active gene promoters. Levels of SETD1A and H3K4me3 are high in NSCs and malignant gliomas, highlighting potentially novel treatment targets. BMP4 increases cell sensitivity to cytotoxic therapies. However, BMP4 effects on H3K4me3 in human NSCs and pediatric HGGs are unknown. In this study, cellular phenotype, levels of H3K4me3 and SETD1A following BMP4 treatment in human NSC and pediatric HGGs are investigated. The results indicate BMP4 promotes differentiation and decreases H3K4me3 and SETD1A levels, as well as H3K4me3 occupancy on stemness and drug resistant genes’ promoters decreased in NSC and pediatric HGGs, respectively. BMP4 also increases chemosensitivity in pediatric HGGs. The results demonstrate that BMP4 differentiates human NSCs and increases chemotherapy sensitivity through regulation of SETD1A by altering H3K4me3 levels at key stemness and drug resistance genes’ promoter. Subsequently, targeting SETD1A concomitantly with conventional chemotherapy may provide novel therapeutic strategies to improve prognosis for children with HGGs.

Chemotherapy is an adjuvant treatment for glioblastomas, however, chemotherapy remains palliative because of the development of multidrug resistance (MDR). Following prolonged chemotherapy, MDR protein 1 (MDR1) and CD133 increase in recurrent glioblastomas. CD133 positive (CD133+) glioma cancer stem-like cells (GCSCs) markedly promote drug resistance and exhibit increased DNA damage repair capability; thus they have a key role in determining tumor chemosensitivity. Although CD133, DNA-dependent protein kinase (DNA-PK), and MDR1 are elevated in CD133+ GCSCs, the relationship among these molecules has not been elucidated. In this study, MDR glioblastoma cell lines were created in response to prolonged doxorubicin chemotherapy. CD133, DNA-PK and MDR1 were markedly elevated in these cells. CD133 and DNA-PK may increase MDR1 via the phosphatidylinositol-3-kinase (PI3K)-Akt signal pathway. PI3K downstream targets Akt and nuclear factor (NF)-κB, which interacts with the MDR1 promoter, were also elevated in these cells. Downregulation of CD133 and DNA-PK by small interfering RNA, or inhibition of PI3K or Akt, decreased Akt, NF-κB and MDR1 expression. The results indicate that CD133 and DNA-PK regulate MDR1 through the PI3K- or Akt-NF-κB signal pathway. Consequently, a novel chemotherapeutic regimen targeting CD133 and DNA-PK in combination with traditional protocols may increase chemotherapeutic efficacy and improve prognosis for individuals who present with glioblastoma.

Environmental endocrine disruptors are implicated as putative contributors to the burgeoning metabolic disease epidemic. Tolylfluanid (TF) is a commonly detected fungicide in Europe, and previous in vitro and ex vivo work has identified it as a potent endocrine disruptor with the capacity to promote adipocyte differentiation and induce adipocytic insulin resistance, effects likely resulting from activation of glucocorticoid receptor signaling. The present study extends these findings to an in vivo mouse model of dietary TF exposure. After 12 weeks of consumption of a normal chow diet supplemented with 100 parts per million TF, mice exhibited increased body weight gain and an increase in total fat mass, with a specific augmentation in visceral adipose depots. This increased adipose accumulation is proposed to occur through a reduction in lipolytic and fatty acid oxidation gene expression. Dietary TF exposure induced glucose intolerance, insulin resistance, and metabolic inflexibility, while also disrupting diurnal rhythms of energy expenditure and food consumption. Adipose tissue endocrine function was also impaired with a reduction in serum adiponectin levels. Moreover, adipocytes from TF-exposed mice exhibited reduced insulin sensitivity, an effect likely mediated through a specific down-regulation of insulin receptor substrate-1 expression, mirroring effects of ex vivo TF exposure. Finally, gene set enrichment analysis revealed an increase in adipose glucocorticoid receptor signaling with TF treatment. Taken together, these findings identify TF as a novel in vivo endocrine disruptor and obesogen in mice, with dietary exposure leading to alterations in energy homeostasis that recapitulate many features of the metabolic syndrome.

Les matières organiques manipulées au cours de la fabrication des mélanges argileux destinés à la fabrication de la poterie sont fréquemment prélevées dans des milieux associés aux activités quotidiennes des artisans, relatives aux pratiques agricoles ou alimentaires. À ce titre, les structures d’origine organique, végétales ou animales, observables dans les produits céramiques cuits constituent une source d’informations paléoéconomiques de premier ordre, en particulier lorsque les données archéobotaniques ou archéozoologiques font défaut, comme c’est le cas pour le site de Koumbi Saleh. Koumbi Saleh (Mauritanie ve-xviie siècles AD) est l’un des plus anciens et plus importants complexes urbains historiques mis au jour en Afrique de l’Ouest. Les fouilles réalisées sur ce site ont livré un matériel céramique caractérisé par une abondante porosité d’origine organique. Afin de préciser l’origine de ces structures, une approche micro-analytique couplant microscopie stéréoscopique et microscopie optique à transmission a été développée. Une analyse des bioclastes associés à ces porosités a également été engagée à titre exploratoire. Enfin, dans le but de renforcer le diagnostic, un référentiel expérimental a été constitué. Les résultats de ce travail de caractérisation suggèrent que les potiers implantés sur le site ont sélectionné des produits secondaires provenant probablement d’herbivores domestiques mais aussi de sous-produits céréaliers. Organic-derived materials used during clay preparation prior to pottery shaping are frequently collected by potters in areas related to their day-to-day activities, such as farming lands, herding areas or places devoted to food processing. Porosity of organic origin in ceramic pastes may thus provide invaluable paleoeconomic information, especially where archaebotanical or archaeozoological data are lacking, as is the case for Koumbi Saleh. Koumbi Saleh (Mauritania, 5e-17e centuries AD) is one of the most ancient and important historical urban sites excavated in Western Africa and has yielded ceramics wih an abundant porosity of organic origin. In order to characterize these organic structures and determine their nature, a microanalytic approach combining stereoscopic and optical transmission microscopy was used, together with a preliminary analysis of the bioclast found in pottery sampless. An experimental reference study was also performed, allowing to strengthen the diagnostic. The results of this work indicate that pottery producers at Koumbi Saleh preferentially selected secondary products related to animal as well as crop exploitation.

States' role in payment as well as coverage will be subject to debate as the administration and the Congress decide how to address the Affordable Care Act (ACA) and otherwise reshape the nation's health policies. Acting as stewards of health care for the entire state population and stimulated by concern about rising costs and federal support under the ACA, the elected and administrative leaders of some states have been using their political influence and authority to improve their state's overall systems of care regardless of who pays the bill. In early 2015 we conducted on-site interviews with key stakeholders in five states to explore their strategies for payment and delivery reform. We found that despite these states' similar goals, differences in their statutory authority and purchasing power, along with their leaders' willingness to use them, significantly influence a state's ability to achieve reform objectives. We caution federal and state policy makers to recognize the reality that state leaders' political desire to exercise stewardship may not be enough to achieve it.