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Abstract The Flood Studies Report (FSR) summer and winter design profiles are a key component of rainfall design guidance in the United Kingdom (UK). We have examined the rainfall profiles of over 70,000 extreme rainfall events with the original FSR profile methodology. This analysis reveals that rainfall profiles change with rainstorm duration but not season, contradicting one of the key assumptions in current UK rainfall design guidance. By using a method that does not artificially generate symmetrical and centred profiles we show that profile shapes are highly variable and strongly related to event duration and magnitude. Short events tend towards front‐loaded profiles, while heavy long‐duration events tend towards centred profile shapes. Finally, manual, automatic and mixed methods of deriving new design profiles for use in the UK were trialled, with consistent results. These could be used to derive new design profiles to supersede the FSR profiles. Notably, peak intensities in observed profiles and trialled summary profiles often exceeded those found in both FSR profiles. We conclude that current design profile guidance for the UK fails to account for the observed variability in event profile shapes and peak intensities and may lead to significant under‐ or over‐design of flood risk management solutions.

Background Tongue-tie can be diagnosed in 3–11% of babies, with some studies reporting almost universal breastfeeding difficulties, and others reporting very few feeding difficulties that relate to the tongue-tie itself, instead noting that incorrect positioning and attachment are the primary reasons behind the observed breastfeeding difficulties and not the tongue-tie itself. The only existing trials of frenotomy are small and underpowered and/or include only very short-term or subjective outcomes. Objective To investigate whether frenotomy is clinically and cost-effective to promote continuation of breastfeeding at 3 months in infants with breastfeeding difficulties diagnosed with tongue-tie. Design A multicentre, unblinded, randomised, parallel group controlled trial. Setting Twelve infant feeding services in the UK. Participants Infants aged up to 10 weeks referred to an infant feeding service (by a parent, midwife or other breastfeeding support service) with breastfeeding difficulties and judged to have tongue-tie. Interventions Infants were randomly allocated to frenotomy with standard breastfeeding support or standard breastfeeding support without frenotomy. Main outcome measures Primary outcome was any breastmilk feeding at 3 months according to maternal self-report. Secondary outcomes included mother’s pain, exclusive breastmilk feeding, exclusive direct breastfeeding, frenotomy, adverse events, maternal anxiety and depression, maternal and infant NHS health-care resource use, cost-effectiveness, and any breastmilk feeding at 6 months of age. Results Between March 2019 and November 2020, 169 infants were randomised, 80 to the frenotomy with breastfeeding support arm and 89 to the breastfeeding support arm from a planned sample size of 870 infants. The trial was stopped in the context of the COVID-19 pandemic due to withdrawal of breastfeeding support services, slow recruitment and crossover between arms. In the frenotomy with breastfeeding support arm 74/80 infants (93%) received their allocated intervention, compared to 23/89 (26%) in the breastfeeding support arm. Primary outcome data were available for 163/169 infants (96%). There was no evidence of a difference between the arms in the rate of breastmilk feeding at 3 months, which was high in both groups (67/76, 88% vs. 75/87, 86%; adjusted risk ratio 1.02, 95% confidence interval 0.90 to 1.16). Adverse events were reported for three infants after surgery [bleeding (n = 1), salivary duct damage (n = 1), accidental cut to the tongue and salivary duct damage (n = 1)]. Cost-effectiveness could not be determined with the information available. Limitations The statistical power of the analysis was extremely limited due to not achieving the target sample size and the high proportion of infants in the breastfeeding support arm who underwent frenotomy. Conclusions This trial does not provide sufficient information to assess whether frenotomy in addition to breastfeeding support improves breastfeeding rates in infants diagnosed with tongue-tie. Future work There is a clear lack of equipoise in the UK concerning the use of frenotomy, however, the effectiveness and cost-effectiveness of the procedure still need to be established. Other study designs will need to be considered to address this objective. Trial registration This trial is registered as ISRCTN 10268851. Funding This project was funded by the National Institute for Health and Care Research (NIHR) Health Technology Assessment Programme (project number 16/143/01) and will be published in full in Health Technology Assessment; Vol. 27, No. 11. See the NIHR Journals Library website for further project information. The funder had no role in study design or data collection, analysis and interpretation. The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health and Social Care. Plain language summary Many mothers and babies experience difficulties in establishing breastfeeding. In some babies it is thought that their difficulties may be linked to a condition called tongue-tie, in which a piece of skin tightly joins the middle part of the underside of the tongue to the base of the baby’s mouth. This can be treated by an operation to divide the tight part/skin in the middle of the underneath of the tongue. We planned to carry out a trial of 870 babies to find out whether an operation together with breastfeeding support helps more mothers and babies with tongue-tie to continue breastfeeding until the baby is 3 months old compared to breastfeeding support on its own and whether the costs were different between the two groups of mothers and babies. We were only able to recruit 169 babies as the trial was stopped because of slow recruitment, changes to services in the COVID-19 pandemic and a high proportion of the babies in the breastfeeding support group going on to have an operation. There were no differences in the rate of breastfeeding at 3 months between the babies in the group who had an operation straightaway and those in the group that had breastfeeding support alone, or had an operation later. More than four in every five babies in both groups were still breastmilk feeding at 3 months. Three babies who had an operation, around 1 in 50 babies, had a complication of the operation (bleeding, scarring or a cut to the tube that makes saliva). Because of the small size of the study, we cannot say whether an operation to divide a tongue-tie along with breastfeeding support helps babies with tongue-tie and breastfeeding difficulties or has different costs. We will need to try different types of research to answer the question. Scientific summary Background Breastfeeding difficulties have been associated with many factors, from a societal to an individual level. Tongue-tie can be diagnosed in 3–11% of babies, with the variation in reported prevalence thought to relate to the use of different diagnostic or severity criteria. Up to half of babies with tongue-tie are reported to have breastfeeding difficulties, but the reported proportion is highly variable. Some studies report almost universal difficulties, and others report very few feeding difficulties that relate to the tongue-tie itself, instead noting that incorrect positioning and attachment are the primary reasons behind the observed breastfeeding difficulties and not the tongue-tie itself. In a UK survey, it was noted that management of tongue-tie in infants with breastfeeding difficulties was therefore highly variable across the country. This is coupled with highly variable provision of breastfeeding support, which can range from minimal to expert and intensive, and using a variety of different models including peer supporter, midwife and health visitor. A Cochrane review identified five prior randomised controlled trials (RCTs) of frenotomy including a total of only 302 infants. The trials are small and underpowered and/or include only very short-term or subjective outcomes, suggesting further robust evidence is needed. Hence there is considerable controversy regarding, not only the diagnosis and clinical significance, but also the management of tongue-tie. Current National Institute for Health and Care Excellence (NICE) guidance allows for the procedure, based on lack of safety concerns, but notes very limited evidence of efficacy. There is therefore a clear need for an assessment of the clinical- and cost-effectiveness of frenotomy for babies diagnosed with tongue-tie in the form of an adequately powered, pragmatic RCT, taking into account the diagnostic controversy and variation in practice. Objective To investigate whether frenotomy is clinically- and cost-effective to promote continuation of breastfeeding at 3 months in infants with breastfeeding difficulties diagnosed with tongue-tie. Methods Study design The FROSTTIE trial was a multicentre, RCT conducted in 12 infant feeding services in England. Participants Inclusion criteria Any infant aged

Climate services must provide robust estimates of future changes to precipitation extremes to inform flood risk management and to assess the resilience of existing urban drainage systems in a changing climate. We use an ensemble of convection-permitting UK climate projections, UKCP Local to estimate return levels of 1–24 h precipitation extremes, combining state-of-the-art climate and spatial statistical modelling. We produce low, central and high precipitation return level uplifts at 5 km resolution across the UK. On average across the UK, the 30y return level of 1 and 24 h precipitation is projected to increase by 30–45% and 20–40% respectively by 2070 under RCP8.5, for central-to-high estimates. Uplifts are consistently higher over the northern UK. For 1 h extremes, uplifts here are higher than the official guidance for some parts of the UK, where guidelines pre-date UKCP Local. These spatially-varying uplifts are given for durations out to 24 h with uplifts lower for longer duration extremes. We provide physical explanations for these spatial patterns and the variation with duration. The new uplifts improve on previous official guidance that used a single value for all durations and across most UK nations, and was based on climate models with poor representation of sub-daily precipitation extremes. The results translate the latest UK climate change projections into tailored climate adaptation information useful and suitable for the water industry and flood risk management stakeholders. This is critical for ensuring we benefit from the latest advances in climate modelling and build a society that is resilient to a changing climate.

BackgroundSelf-administered paper or electronic surveys can create accessibility issues for people with language barriers and limited literacy, whereas face-to-face interviews can create privacy issues and give rise to reporting biases, particularly in the context of sensitive subject matters. An audio computer-assisted self-interview (ACASI) offers an alternative mode of survey administration, and its use has been tested against other survey modes to determine whether the presence of a background narration helps overcome literacy and privacy issues. There are still gaps with the ACASI survey administration because audio narration alone does not assist respondents with limited literacy in choosing response options. To overcome literacy issues, a few studies have used illustrated pictures for a limited number of response options. ObjectiveThis study aimed to illustrate all the questions and response options in an ACASI application. This research is part of a larger study comparing different modes of survey administration (ACASI, face-to-face interviews, and self-administered paper surveys) to collect data on hepatitis B knowledge, attitudes, and practices among the Myanmar-born community in Perth, Australia. This study describes the 2-phase process of developing a web-based ACASI application using illustrated pictures. MethodsThe first phase was the preparation of the ACASI elements, such as questionnaire, pictures, brief descriptions of response options, and audio files. Each element was pretested on 20 participants from the target population. The second phase involved synchronizing all the elements into the web-based ACASI application and adapting the application features, in particular, autoplay audio and illustrated pictures. The preprototype survey application was tested for user acceptance on 5 participants from the target population, resulting in minor adjustments to the display and arrangement of response options. ResultsAfter a 12-month development process, the prototype ACASI application with illustrated pictures was fully functional for electronic survey administration and secure data storage and export. ConclusionsPretesting each element separately was a useful approach because it saved time to reprogram the application at a later stage. Future studies should also consider the participatory development of pictures and visual design of user interfaces. This picture-assisted ACASI survey administration mode can be further developed and used to collect sensitive information from populations that are usually marginalized because of literacy and language barriers.

Abstract The risk of surface water flooding (SWF) in England is already high and its frequency and severity is projected to increase in the future. SWF generally occurs due to intense, highly localised rainfall, which is challenging to forecast with sufficient accuracy to take proactive action ahead of flood events. Being able to manage the risk effectively lies in improved rainfall and flood forecast products, better communication of uncertainty and building the capacity of local responders. This study utilises state‐of‐the‐art high‐resolution ensemble rainfall forecasts and hydraulic modelling tools alongside a novel post‐processing method to develop and trial new SWF forecast products within an incident workshop attended by forecast producers and regional forecast users. Twenty‐two of 24 workshop participants reported that the new information would be useful to their organisation but more product development and training in its interpretation is required. Specific recommendations to improve SWF forecast provision include increased support for local government through a single government organisation responsible for SWF, making more use of existing static SWF mapping in a real‐time context and employing the process of user‐based consultation, as outlined in this study, to guide the future development of future SWF forecast information and processes.

In this study the transfer characteristics of mercury (Hg) from a wide range of Chinese soils to corn grain (cultivar Zhengdan 958) were investigated. Prediction models were developed for determining the Hg bioconcentration factor (BCF) of Zhengdan 958 from soil, including the soil properties, such as pH, organic matter (OM) concentration, cation exchange capacity (CEC), total nitrogen concentration (TN), total phosphorus concentration (TP), total potassium concentration (TK), and total Hg concentration (THg), using multiple stepwise regression analysis. These prediction models were applied to other non-model corn cultivars using a cross-species extrapolation approach. The results indicated that the soil pH was the most important factor associated with the transfer of Hg from soil to corn grain. Hg bioaccumulation in corn grain increased with the decreasing pH. No significant differences were found between two prediction models derived from different rates of Hg applied to the soil as HgCl2. The prediction models established in this study can be applied to other non-model corn cultivars and are useful for predicting Hg bioconcentration in corn grain and assessing the ecological risk of Hg in different soils.

Background: Vascular smooth muscle cells (VSMCs) migration is a critical process during human uterine spiral artery (SpA) remodeling and a successful pregnancy. Extravillous trophoblast cells (EVT) interact with VSMC and enhance their migration, however, the mechanisms by which EVT remodel SpA remain to be fully elucidated. We hypothesize that exosomes released from EVT promote VSMC migration.Methods: JEG-3 and HTR-8/SVneo cell lines were used as models for EVT. Cells were cultured at 37 0C and humidified under an atmosphere of 5% CO2-balanced N2 to obtain 8% O2. Cell-conditioned media were collected and exosomes (exo-JEG-3 and exo- HTR-8/SVneo) isolated by differential and buoyant density centrifugation. The effects of exo-EVT on VSMC migration were established using a real-time, live-cell imaging system (Incucyte™). Exosomal proteins where identified by mass spectrometry and submitted to bioinformatic pathway analysis (Ingenuity software) .Results: HTR-8/SVneo cells were significantly more (~30%) invasive than JEG-3 cells. HTR-8/SVneo cells released 2.6-fold more exosomes (6.39 x 108 ± 2.5 x108 particles/106 cells) compared to JEG-3 (2.86 x 108 ± 0.78 x108 particles/106 cells). VSMC migration was significantly increased in the presence of exo-JEG-3 and exo-HTR-8/SVneo compared to control (-exosomes) (21.83 ± 0.49 h and 15.57 ± 0.32, respectively, versus control 25.09 ± 0.58 h, p

Specialised cell types of trophoblast cells form the placenta in which each cell type has particular properties of proliferation and invasion. The placenta sustains the growth of the fetus throughout pregnancy and any aberrant trophoblast differentiation or invasion potentially affects the future health of the child and adult. Recently, the field of epigenetics has been applied to understand differentiation of trophoblast lineages and embryonic stem cells, from fertilisation of the oocyte onward. Each trophoblast cell-type has a distinctive epigenetic profile and we will concentrate on the epigenetic mechanism of DNA methyltransferases and TETs that regulate DNA methylation. Environmental factors affecting the mother potentially regulate the DNA methyltransferases in trophoblasts, and so do steroid hormones, cell cycle regulators, such as p53, and cytokines, especially interlukin-1β. There are interesting questions of why trophoblast genomes are globally hypomethylated yet specific genes can be suppressed by hypermethylation (in general, tumour suppressor genes, such as E-cadherin) and how invasive cell-types are liable to have condensed chromatin, as in metastatic cancer cells. Future work will attempt to understand the interactive nature of all epigenetic mechanisms together and their effect on the complex biological system of trophoblast differentiation and invasion in normal as well as pathological conditions.

Sympathetic activation in subjects with the metabolic syndrome (MS) plays a role in the pathogenesis of cardiovascular disease development. Diet-induced weight loss decreases sympathetic outflow. However the mechanisms that account for sympathetic inhibition are not known. We sought to provide a detailed description of the sympathetic response to diet by analyzing the firing behavior of single-unit sympathetic nerve fibres. Fourteen subjects (57±2 years, 9 men, 5 females) fulfilling ATP III criteria for the MS underwent a 3-month low calorie diet. Metabolic profile, hemodynamic parameters and multi-unit and single unit muscle sympathetic nerve activity (MSNA, microneurography) were assessed prior to and at the end of the diet. Patients’ weight dropped from 96±4 to 88±3 kg (P

The development and growth of information technology in the past decade has changed the face of communication at both a personal and an organisational level. As young people and practitioners become more familiar with its applications, it seems pertinent to think about how it can be applied more widely in relation to health and wellbeing in residential child care settings. An increasing number of local authorities are introducing audio computer assisted self-interviewing (A-CASI) as a method for improving communication with young people in their care. Self-completion methods in general are viewed as advantageous, in comparison to other approaches, in terms of being cheaper and quicker to administer and also in terms of minimizing the under-reporting of issues that could be sensitive (De Vaus, 1996). A-CASI approaches have been identified as being of particular benefit in conducting research with particular groups, such as children and young people and have also been associated with aiding literacy difficulties, with an enhanced sense of privacy and with increased disclosure of sensitive information (De Leeuw et al., 1997; Borgers et al., 2000; Borgers et al., 2004). In addition, its use is thought to decrease respondent error or fatigue and allows the relatively easy use of more complicated questionnaires, providing richer data than other self-completion approaches (Tourangeau and Smith, 1996).

Abstract Background There are concerns that pre-operative psychological distress might be associated with reduced patient satisfaction after total hip replacement (THR). Methods We investigated this in a multi-centre prospective study between January 1999 and January 2002. We dichotomised the patients into the mentally distressed (MHS ≤ 56) and the not mentally distressed (MHS > 56) groups based on their pre-operative Mental Health Score (MHS) of SF36. Results 448 patients (340 not distressed and 108 distressed) completed the patient satisfaction survey. Patient satisfaction rate at five year was 96.66% (415/448). There was no difference in patient satisfaction or willingness to have the surgery between the two groups. None of pre-operative variables predicted five year patient satisfaction in logistic regression. Conclusions Patient satisfaction after surgery may not be adversely affected by pre-operative psychological distress.

Abstract Background NET1, a RhoA guanine exchange factor, is up-regulated in gastric cancer (GC) tissue and drives the invasive phenotype of this disease. In this study, we aimed to determine the role of NET1 in GC by monitoring the proliferation, motility and invasion of GC cells in which NET1 has been stably knocked down. Additionally, we aimed to determine NET1-dependent transcriptomic events that occur in GC. Methods An in vitro model of stable knockdown of NET1 was achieved in AGS human gastric adenocarcinoma cells via lentiviral mediated transduction of short-hairpin (sh) RNA targeting NET1. Knockdown was assessed using quantitative PCR. Cell proliferation was assessed using an MTS assay and cell migration was assessed using a wound healing scratch assay. Cell invasion was assessed using a transwell matrigel invasion assay. Gene expression profiles were examined using affymetrix oligonucleotide U133A expression arrays. A student's t test was used to determine changes of statistical significance. Results GC cells were transduced with NET1 shRNA resulting in a 97% reduction in NET1 mRNA (p < 0.0001). NET1 knockdown significantly reduced the invasion and migration of GC cells by 94% (p < 0.05) and 24% (p < 0.001) respectively, while cell proliferation was not significantly altered following NET1 knockdown. Microarray analysis was performed on non-target and knockdown cell lines, treated with and without 10 μM lysophosphatidic acid (LPA) allowing us to identify NET1-dependent, LPA-dependent and NET1-mediated LPA-induced gene transcription. Differential gene expression was confirmed by quantitative PCR. Shortlisted NET1-dependent genes included STAT1, TSPAN1, TGFBi and CCL5 all of which were downregulatd upon NET1 downregulation. Shortlisted LPA-dependent genes included EGFR and PPARD where EGFR was upregulated and PPARD was downregulated upon LPA stimulation. Shortlisted NET1 and LPA dependent genes included IGFR1 and PIP5K3. These LPA induced genes were downregulated in NET1 knockdown cells. Conclusions NET1 plays an important role in GC cell migration and invasion, key aspects of GC progression. Furthermore, the gene expression profile further elucidates the molecular mechanisms underpinning NET1-mediated aggressive GC cell behaviour.

Abstract Background The Wnt/β-catenin pathway is a major signaling cascade in bone biology, playing a key role in bone development and remodeling. The objectives of this study were firstly, to determine the effects of dexamethasone exposure on Wnt/β-catenin signaling at an intracellular and transcriptional level, and secondly, to assess the phenotypic effects of silencing the Wnt antagonist, Dickkopf-1 (Dkk1) in the setting of dexamethasone exposure. Methods Primary human osteoblasts were exposed in vitro to 10-8 M dexamethasone over a 72 h time course. The phenotypic marker of osteoblast differentiation was analyzed was alkaline phosphatase activity. Intracellular β-catenin trafficking was assessed using immunoflourescence staining and TCF/LEF mediated transcription was analyzed using a Wnt luciferase reporter assay. Dkk1 expression was silenced using small interfering RNA (siRNA). Results Primary human osteoblasts exposed to dexamethasone displayed a significant reductions in alkaline phosphatase activity over a 72 h time course. Immunoflourescence analaysis of β-catenin localization demonstrated a significant reduction in intracytosolic and intranuclear β-catenin in response to dexamethasone exposure. These changes were associated with a reduction of TCF/LEF mediated transcription. Silencing Dkk1 expression in primary human osteoblasts exposed to dexamethasone resulted in an increase in alkaline phosphatase activity when compared to scrambled control. Conclusions Wnt/β-catenin signaling plays a key role in regulating glucocorticoid-induced osteoporosis in vitro. Silencing Dkk1 expression rescues dexamethasone-induced suppression of primary human osteoblast differentiation. Targeting of the Wnt/β-catenin signaling pathway offers an exciting opportunity to develop novel anabolic bone agents to treat osteoporosis and disorders of bone mass.

Abstract Introduction We have previously reported that Myeov (MYEloma OVerexpressed gene) expression is enhanced in colorectal cancer (CRC) and that it promotes CRC cell proliferation and invasion. The role of Myeov in CRC migration is unclear. ProstaglandinE2 (PGE 2) is a known factor in promoting CRC carcinogenesis. The role of PGE 2 in modulating Myeov expression has also not been defined. Aim To assess the role of Myeov expression in CRC cell migration and to evaluate the role of PGE 2 in Myeov bioactivity. Methods siRNA mediated Myeov knockdown was achieved in T84 CRC cells. Knockdown was assessed using quantitative real time PCR. The effect of knockdown on CRC cell migration was assessed using a scratch wound healing assay. Separately, T84 cells were treated with PGE 2 (0.00025 μ M, 0.1 μ M and 1 μ M) from 30 min to 3 hours and the effect on Myeov gene expression was assessed using real time PCR. Results Myeov knockdown resulted in a significant reduction in CRC cell migration, observable as early as 12 hours (P < 0.05) with a 39% reduction compared to control at 36 hours (p < 0.01). Myeov expression was enhanced after treatment with PGE 2, with the greatest effect seen at 60 mins for all 3 PGE 2 doses. This response was dose dependent with a 290%, 550% & 1,000% increase in Myeov expression for 0.00025 μ M, 0.1 μ M and 1 μ M PGE 2 respectively. Conclusion In addition to promoting CRC proliferation and invasion, our findings indicate that Myeov stimulates CRC cell migration, and its expression may be PGE 2 dependant.

Abstract Background Anaesthetic pre-registration house officer posts have been available since 1997. With the change to postgraduate medical training introduced in 2005, these posts have become vital building blocks for Foundation Programmes. Discussion We debate the skills that new Foundation Programme doctors in such posts should be taught, particularly whether administration of an anaesthetic holds an important place. The opinion of college tutors prior to the institution of the foundation programme is included. These were obtained from a postal questionnaire. Summary We maintain that teaching how to administer an anaesthetic remains an important learning objective and something that should be actively pursued.

Abstract Efforts aimed at deciphering the molecular basis of complex disease are underpinned by the availability of high throughput strategies for the identification of biomolecules that drive the disease process. The completion of the human genome-sequencing project, coupled to major technological developments, has afforded investigators myriad opportunities for multidimensional analysis of biological systems. Nowhere has this research explosion been more evident than in the field of transcriptomics. Affordable access and availability to the technology that supports such investigations has led to a significant increase in the amount of data generated. As most biological distinctions are now observed at a genomic level, a large amount of expression information is now openly available via public databases. Furthermore, numerous computational based methods have been developed to harness the power of these data. In this review we provide a brief overview of in silico methodologies for the analysis of differential gene expression such as Serial Analysis of Gene Expression and Digital Differential Display. The performance of these strategies, at both an operational and result/output level is assessed and compared. The key considerations that must be made when completing an in silico expression analysis are also presented as a roadmap to facilitate biologists. Furthermore, to highlight the importance of these in silico methodologies in contemporary biomedical research, examples of current studies using these approaches are discussed. The overriding goal of this review is to present the scientific community with a critical overview of these strategies, so that they can be effectively added to the tool box of biomedical researchers focused on identifying the molecular mechanisms of disease.

Abstract Background Osteoporosis, a disease of decreased bone mineral density represents a significant and growing burden in the western world. Aging population structure and therapeutic use of glucocorticoids have contributed in no small way to the increase in the incidence of this disease. Despite substantial investigative efforts over the last number of years the exact molecular mechanism underpinning the initiation and progression of osteoporosis remain to be elucidated. This has meant that no significant advances in therapeutic strategies have emerged, with joint replacement surgery being the mainstay of treatment. Methods In this study we have used an integrated genomics profiling and computational biology based strategy to identify the key osteoblast genes and gene clusters whose expression is altered in response to dexamethasone exposure. Primary human osteoblasts were exposed to dexamethasone in vitro and microarray based transcriptome profiling completed. Results These studies identified approximately 500 osteoblast genes whose expression was altered. Functional characterization of the transcriptome identified developmental networks as being reactivated with 106 development associated genes found to be differentially regulated. Pathway reconstruction revealed coordinate alteration of members of the WNT signaling pathway, including frizzled-2, frizzled-7, DKK1 and WNT5B, whose differential expression in this setting was confirmed by real time PCR. Conclusion The WNT pathway is a key regulator of skeletogenesis as well as differentiation of bone cells. Reactivation of this pathway may lead to altered osteoblast activity resulting in decreased bone mineral density, the pathological hallmark of osteoporosis. The data herein lend weight to the hypothesis that alterations in developmental pathways drive the initiation and progression of osteoporosis.

Exact analytical expressions for the spatial impulse response are available for certain transducer geometries. These exact expressions for the spatial impulse response, which are only available for lossless media, analytically evaluate the Rayleigh integral to describe the effect of diffraction in the time domain. To extend the concept of the spatial impulse response by including the effect of power law attenuation in a lossy medium, time-domain Green's functions for the Power Law Wave Equation, which are expressed in terms of stable probability density functions, are computed numerically and superposed. Numerical validations demonstrate that the lossy spatial impulse for a circular piston converges to the analytical lossless spatial impulse response as the value of the attenuation constant grows small. The lossy spatial impulse response is then evaluated in different spatial locations for four specific values of the power law exponent using several different values for the attenuation constant. As the attenuation constant or the distance from the source increases, the amplitude decreases while an increase in temporal broadening is observed. The sharp edges that appear in the time-limited lossless impulse response are replaced by increasingly smooth curves in the lossy impulse response, which decays slowly as a function of time.

The Mars Desert Research Station (MDRS) is a Mars-simulation campus set in a Martian planetary analogue in southern Utah. Despite a long history of astrobiology research, collections-based taxonomic inventories of the macro-level biodiversity around the station are relatively new. This study serves to add to the initial vascular plant list published for the station in 2016, where 39 species were recorded for MDRS. Here we report 40 new species, two new taxa recorded only to genus and two species re-identified from our 2016 fieldwork, bringing the total number of taxa in the "Martian" flora to 79 species and two taxa recorded to genus., (Paul C. Sokoloff, David A. Murray, Samantha R.M. McBeth, Michael G. Irvine, Shannon M. Rupert.)

Currently, in a number of public and semi-public forums in Barbados, the idea of 'sexual rights' is being discussed and debated. However, different meanings are attached to 'rights'. This paper examines how these meanings demonstrate that different interpretations of sexuality, society, and morality are circulating through Barbados today. It also addresses whether or not sexual rights discourses are the best way to advocate for social justice or bring about changes to socio-sexual attitudes in the Caribbean. It is argued that framing justice and equality through rights talk may have deleterious effects for its advocates, as there is no 'clear' or transparent universality as to what rights means. It is suggested that it may be more efficacious for groups who are stigmatized based on sexual orientation to develop vernacular strategies with values and/or logics stressing elements of justice, equality, dignity and respect for personhood, which include but also move beyond sexual orientation as a principal identification.

Respiratory infections, including influenza in humans, are often accompanied by a hepatitis that is usually mild and self-limiting. The mechanism of this kind of liver damage is not well understood. In the present study, we show that influenza-associated hepatitis occurs due to the formation of inflammatory foci that include apoptotic hepatocytes, antigen-specific CD8(+) T cells, and Kupffer cells. Serum aminotransaminase levels were elevated, and both the histological and serum enzyme markers of hepatitis were increased in secondary influenza infection, consistent with a primary role for antigen-specific T cells in the pathogenesis. No virus could be detected in the liver, making this a pure example of "collateral damage" of the liver. Notably, removal of the Kupffer cells prevented the hepatitis. Such hepatic collateral damage may be a general consequence of expanding CD8(+) T-cell populations during many extrahepatic viral infections, yielding important implications for liver pathobiology.

Background: Transgenic TCR mice are often used experimentally as a source of T cells of a defined specificity. One of the most widely used transgenic TCR models is the OT-1 transgenic mouse in which the CD8+ T cells express a TCR specific for the SIINFEKL peptide of ovalbumin presented on kb. Although OT-1 CD8+ can be used in a variety of different experimental settings, we principally employ adoptive transfer and peptide-driven expansion of OT-1 cells in order to explore the distribution and fate of these antigen-specific OT-1 T cells. We set out to develop a quantitative PCR assay for OT-1 cells in order to assess the distribution of OT-1 CD8+ T cells in tissues that are either intrinsically difficult to dissociate for flow cytometric analysis or rendered incompatible with flow cytometric analysis through freezing or fixation., Results: We show excellent correlation between flow cytometric assessment of OT-1 cells and OT-1 signal by qPCR assays in cell dilutions as well as in in vivo adoptive transfer experiments. We also demonstrate that qPCR can be performed from archival formalin-fixed paraffin-embedded tissue sections. In addition, the non-quantitative PCR using the OT-1-specific primers without the real-time probe is a valuable tool for OT-1 genotyping, obviating the need for peripheral blood collection and subsequent flow cytometric analysis., Conclusion: An OT-1 specific qPCR assay has been developed to quantify adoptively transferred OT-1 cells. OT-1 qPCR to determine cell signal is a valuable adjunct to the standard flow cytometric analysis of OT-1 cell number, particularly in experimental settings where tissue disaggregation is not desirable or in tissues which are not readily disassociated.

Lipopolysaccharide (LPS) from Porphyromonas gingivalis prevented apoptosis of HL60-derived neutrophils, which could not be restored upon the addition of interleukin-10. Signaling of P. gingivalis LPS through Toll-like receptor 2 (TLR2), not TLR4, may account for the inhibiting effect of P. gingivalis LPS on apoptosis and provide a mechanism for the development of destructive periodontal inflammation.

Measurement characteristics of two constant velocity dynamometers (Cybex II, Lumex, Inc., NY) were evaluated at 11 selected speeds (0 to 5 rad X s-1) under various conditions of inert gravitational loading. A linear accelerometer was affixed to the lever arm. Speed settings were standardized percentages of each machine's maximum tachometer feedback voltage (0 to 100% Vfb max). Regression of Cybex transducer-observed voltages (V0) on true, i.e., expected, torques (Me) revealed that, for any static loading condition or angle, the measurement system was essentially linear and valid, although greater variance was evident for smaller torques. However, different calibrations, i.e., regression lines, were required for different speed settings. The error in predicting Me from V0 increased substantially with increased % Vfb max. Methodologies which assume constant velocity dynamic loading induce questionable, often invalid, results and inferences. Dynamic V0 represents both gravitational and inertial components. Considerable fluctuations in lever arm instantaneous velocity were ascribed to the combined influence of load and % Vfb max upon the servomotor. After computing inertia-corrected Me, calibration variability and prediction error were markedly reduced. Unexplained variability may be due to poor proximity of the Cybex transducer to the force application point. We concluded that: Cybex II should be calibrated only statically; use of an optimal filter will avoid false estimates of joint torque; inertial corrections must be applied in order to minimize potentially serious errors due to system angular acceleration; and reported muscle function relationships might be usefully re-assessed in view of these findings.

The purpose of this investigation was to implement an optimal filtering strategy for processing in vivo dynamometric data. The validity of employing commonly accepted analog smoothing methods was also appraised. An inert gravitational model was used to assess the filtering requirements of two Cybex II constant velocity dynamometers at 10 pre-set speeds with three selected loads. Speed settings were recorded as percentages of the servomechanism's maximum tachometer feedback voltage (10 to 100% Vfb max). Spectral analyses of unsmoothed torque and associated angular displacement curves, followed by optimized low-pass digital filtering, revealed the presence of two superimposed contaminating influences: a damped oscillation, representing successive sudden braking and releasing of the servomechanism control system; a relatively stationary oscillatory series, which was attributed to the Cybex motor. The optimal cutoff frequency for any data set was principally a positive function of % Vfb max. This association was represented for each machine by a different, but reliable, third order least-squares polynomial, which could be used to accurately predict the correct smoothing required for any speed setting. Unacceptable errors may be induced, especially when measuring peak torques, if data are inappropriately filtered. Over-smoothing disguises inertial artefacts. The use of Cybex recorder damping settings should be discouraged. Optimal filtering is a minimal requirement of valid data processing.