The intracellular plasma membrane-connected compartment in the assembly of HIV-1 in human macrophages

Background In HIV-infected macrophages, newly formed progeny virus particles accumulate in intracellular plasma membrane-connected compartments (IPMCs). Although the virus is usually seen in these compartments, it is unclear whether HIV assembly is specifically targeted to IPMCs or whether some viruses may also form at the cell surface but are not detected, as particles budding from the latter site will be released into the medium. Results To investigate the fidelity of HIV-1 targeting to IPMCs compared to the cell surface directly, we generated mutants defective in recruitment of the Endosomal Sorting Complexes Required for Transport (ESCRT) proteins required for virus scission. For mutants unable to bind the ESCRT-I component Tsg101, HIV release was inhibited and light and electron microscopy revealed that budding was arrested. When expressed in human monocyte-derived macrophages (MDM), these mutants formed budding-arrested, immature particles at their assembly sites, allowing us to capture virtually all of the virus budding events. A detailed morphological analysis of the distribution of the arrested viruses by immunofluorescence staining and confocal microscopy, and by electron microscopy, demonstrated that HIV assembly in MDMs is targeted primarily to IPMCs, with fewer than 5 % of budding events seen at the cell surface. Morphometric analysis of the relative membrane areas at the cell surface and IPMCs confirmed a large enrichment of virus assembly events in IPMCs. Serial block-face scanning electron microscopy of macrophages infected with a budding-defective HIV mutant revealed high-resolution 3D views of the complex organisation of IPMCs, with in excess of 15,000 associated HIV budding sites, and multiple connections between IPMCs and the cell surface

A qualitative study exploring midwives’ perceptions and knowledge of maternal obesity. Reflecting on their experiences of providing healthy eating and weight management advice to pregnant women.

Midwives are responsible for providing advice regarding the complex issues of healthy eating and weight management during pregnancy. This study utilised an inductive data-driven thematic approach in order to determine midwives’ perceptions, knowledge and experiences of providing healthy eating and weight management advice to pregnant women. Semi-structured interviews with 17 midwives were transcribed verbatim and data subjected to thematic analysis. The findings offer insight into the challenges facing midwives in their role trying to promote healthy eating and appropriate weight management to pregnant women. Three core themes were identified: 1) “If they eat healthily it will bring their weight down”: Midwives Misunderstood; 2) “I don’t think we are experienced enough”: Midwives Lack Resources and Expertise, and 3) “BMI of 32 wouldn’t bother me”: Midwives Normalised Obesity. The midwives recognised the importance of providing healthy eating advice to pregnant women and the health risks associated with poor diet and obesity. However, they reported the normalisation of obesity in pregnant women and suggested that this, together with their high workload and lack of expertise, explained the reasons why systematic advice was not in standard antenatal care. In addition, the current lack of UK clinical guidance, and thus possibly lack of clinical leadership, are also preventing delivery of tailored advice. Implementation literature on understanding the barriers to optimal healthcare delivery and informing clinical practice through research evidence needs to be further investigated in this field. This study has recommendations for policy makers, commissioners, service providers and midwives. Keywords: midwives, pregnancy, obesity, healthy eating, qualitative, self-efficac

3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

The processes of life take place in multiple dimensions, but imaging these processes in even three dimensions is challenging. Here, we describe a workflow for 3D correlative light and electron microscopy (CLEM) of cell monolayers using fluorescence microscopy to identify and follow biological events, combined with serial blockface scanning electron microscopy to analyse the underlying ultrastructure. The workflow encompasses all steps from cell culture to sample processing, imaging strategy, and 3D image processing and analysis. We demonstrate successful application of the workflow to three studies, each aiming to better understand complex and dynamic biological processes, including bacterial and viral infections of cultured cells and formation of entotic cell-in-cell structures commonly observed in tumours. Our workflow revealed new insight into the replicative niche of Mycobacterium tuberculosis in primary human lymphatic endothelial cells, HIV-1 in human monocytederived macrophages, and the composition of the entotic vacuole. The broad application of this 3D CLEM technique will make it a useful addition to the correlative imaging toolbox for biomedical research

Dairy consumption and cardiometabolic diseases: systematic review and updated meta-analyses of prospective cohort studies

Purpose of Review Dairy products contain both beneficial and harmful nutrients in relation to cardiometabolic diseases. Here, we provide the latest scientific evidence regarding the relationship between dairy products and cardiometabolic diseases by reviewing the literature and updating meta-analyses of observational studies. Recent Findings We updated our previous meta-analyses of cohort studies on type 2 diabetes, coronary heart disease (CHD), and stroke with nine studies and confirmed previous results. Total dairy and low-fat dairy (per 200 g/d) were inversely associated with a 3–4% lower risk of diabetes. Yogurt was non-linearly inversely associatedwith diabetes (RR = 0.86, 95%CI: 0.83–0.90 at 80 g/ d). Total dairy and milk were not associated with CHD (RR~1.0). An increment of 200 g of daily milk intake was associated with an 8% lower risk of stroke. Summary The latest scientific evidence confirmed neutral or beneficial associations between dairy products and risk of cardiometabolic diseases

Influence of pain location and hand dominance on scapular kinematics and EMG activities: an exploratory study

<p>Abstract</p> <p>Background</p> <p>Assessment of three-dimensional kinematics and electromyography (EMG) activities is common in patients with chronic neck pain. However, the effect of hand dominance and neck pain location on the measurement of movement and EMG characteristics is still unclear. Therefore, the purpose of this study was to investigate the effect of neck pain location and arm dominance on the scapular kinematics and muscle EMG activities in patients with chronic neck pain.</p> <p>Methods</p> <p>Thirty subjects (10 males, 20 females; mean age (sd): 38 (11.9) years) with chronic neck pain for more than 3 months were recruited. The scapular kinematics and EMG activity of the upper trapezius and sternocleidomastoid muscles were measured during the bilateral arm elevation task. The three-way repeated measures ANOVA was used to examine the effect of neck pain location and hand dominance on the measurement of kinematics and EMG muscle activities.</p> <p>Results</p> <p>The movement of scapular posterior tilt was significantly influenced by arm dominance (P = 0.001) and by the interaction of arm dominance and elevation angle (P = 0.002). The movement of scapular upward/downward rotation was affected by the interaction of arm dominance and elevation angle (P = 0.02). The location of pain did not show any significant influence on the scapular movement and muscle activities.</p> <p>Conclusions</p> <p>Hand dominance could have an influence on the scapular kinematics, which should be taken into consideration when describing and comparing neuromuscular characteristics in individuals with chronic neck pain.</p

The achievement of food and nutrition security in South Asia is deeply gendered

Women form an integral part of the agricultural sector, and in much of South Asia women make up a majority of the agricultural workforce, often compelled to work in order to meet their families’ basic needs. While their contributions are recognised as central to the food and nutrition security of households and communities, their work is not recognized or supported adequately by public policy and social institutions. Women continue to face inequality across key development indicators including health, education, and nutrition; discriminatory laws; and high levels of precarity in terms of income, employment conditions, safety and wellbeing. Social structures that promote gender inequality and inhibit the agency of women contribute to the South Asian enigma – the persistence of undernutrition despite economic growth – and must be addressed to achieve food and nutrition security

The effect of premature extraction of primary teeth on the subsequent need for orthodontic treatment.

AIM: To investigate if premature extraction of primary teeth was associated with orthodontic need in the permanent dentition. STUDY DESIGN: This was a case-control study based on retrospective dental records. METHODS: As part of NHS (UK) Dental Epidemiology Programme a sample of 366, 12-year-old children from Bradford and Airedale were examined. The survey collected data on patient demographics, dental health status including orthodontic need. Data linkage was undertaken for those children participating in the NHS Dental Epidemiology Programme who had previously accessed the local Salaried Dental Service (SDS). For these children, retrospective dental information was collected about premature extraction of primary teeth. RESULTS: From the 366 children who were surveyed, 116 children had received treatment at the local SDS in the past. Significantly more children from ethnic minorities, low socioeconomic backgrounds and high caries rate (p < 0.001) were seen in the SDS. For the 107 children who attended SDS, an increased total number of primary teeth extractions was positively associated with orthodontic need (odds ratio:1.18, CI -1.01 to 1.37). STATISCTICS: Multilevel modelling was undertaken to identify variables associated with orthodontic need. CONCLUSIONS: In the study group, orthodontic need was significantly associated with the number of primary teeth extracted

Adherence to antidepressant therapy for major depressive patients in a psychiatric hospital in Thailand

Poor adherence to antidepressant therapy is an important barrier to the effective management of major depressive disorder. This study aims to quantify the adherence rate to antidepressant treatment and to determine the pattern of prescriptions of depressed patients in a psychiatric institute in Thailand.This retrospective study used electronic pharmacy data of outpatients aged 15 or older, with a new diagnosis of major depression who received at least one prescription of antidepressants between August 2005 and September 2008. The medication possession ratio (MPR) was used to measure adherence over a 6 month period.1,058 were eligible for study inclusion. The overall adherence (MPR > 80%) in those attending this facility at least twice was 41% but if we assume that all patients who attended only once were non-adherent, adherence may be as low as 23%. Fluoxetine was the most commonly prescribed drug followed by TCAs. A large proportion of cases received more than one drug during one visit or was switched from one drug to another (39%).Adherence to antidepressant therapy for treatment of major depression in Thailand is rather low compared to results of adherence from elsewhere

An iterative strategy combining biophysical criteria and duration hidden Markov models for structural predictions of Chlamydia trachomatis σ66 promoters

<p>Abstract</p> <p>Background</p> <p>Promoter identification is a first step in the quest to explain gene regulation in bacteria. It has been demonstrated that the initiation of bacterial transcription depends upon the stability and topology of DNA in the promoter region as well as the binding affinity between the RNA polymerase σ-factor and promoter. However, promoter prediction algorithms to date have not explicitly used an ensemble of these factors as predictors. In addition, most promoter models have been trained on data from <it>Escherichia coli</it>. Although it has been shown that transcriptional mechanisms are similar among various bacteria, it is quite possible that the differences between <it>Escherichia coli </it>and <it>Chlamydia trachomatis </it>are large enough to recommend an organism-specific modeling effort.</p> <p>Results</p> <p>Here we present an iterative stochastic model building procedure that combines such biophysical metrics as DNA stability, curvature, twist and stress-induced DNA duplex destabilization along with duration hidden Markov model parameters to model <it>Chlamydia trachomatis </it>σ⁶⁶promoters from 29 experimentally verified sequences. Initially, iterative duration hidden Markov modeling of the training set sequences provides a scoring algorithm for <it>Chlamydia trachomatis </it>RNA polymerase σ⁶⁶/DNA binding. Subsequently, an iterative application of Stepwise Binary Logistic Regression selects multiple promoter predictors and deletes/replaces training set sequences to determine an optimal training set. The resulting model predicts the final training set with a high degree of accuracy and provides insights into the structure of the promoter region. Model based genome-wide predictions are provided so that optimal promoter candidates can be experimentally evaluated, and refined models developed. Co-predictions with three other algorithms are also supplied to enhance reliability.</p> <p>Conclusion</p> <p>This strategy and resulting model support the conjecture that DNA biophysical properties, along with RNA polymerase σ-factor/DNA binding collaboratively, contribute to a sequence's ability to promote transcription. This work provides a baseline model that can evolve as new <it>Chlamydia trachomatis </it>σ⁶⁶promoters are identified with assistance from the provided genome-wide predictions. The proposed methodology is ideal for organisms with few identified promoters and relatively small genomes.</p