An Evaluation of the Precision of Measurement of Ryff's Psychological Well-Being Scales in a Population Sample.
The aim of this study is to assess the effective measurement range of Ryff's Psychological Well-being scales (PWB). It applies normal ogive item response theory (IRT) methodology using factor analysis procedures for ordinal data based on a limited information estimation approach. The data come from a sample of 1,179 women participating in a midlife follow-up of a national birth cohort study in the UK. The PWB scales incorporate six dimensions: autonomy, positive relations with others, environmental mastery, personal growth, purpose in life and self-acceptance. Scale information functions were calculated to derive standard errors of measurement for estimated scores on each dimension. Construct variance was distinguished from method variance by inclusion of method factors from item wording (positive versus negative). Our IRT analysis revealed that the PWB measures well-being most accurately in the middle range of the score distribution, i.e. for women with average well-being. Score precision diminished at higher levels of well-being, and low well-being was measured more reliably than high well-being. A second-order well-being factor loaded by four of the dimensions achieved higher measurement precision and greater score accuracy across a wider range than any individual dimension. Future development of well-being scales should be designed to include items that are able to discriminate at high levels of well-being.


Applications of cellular fatty acid analysis.
More than ever, new technology is having an impact on the tools of clinical microbiologists. The analysis of cellular fatty acids by gas-liquid chromatography (GLC) has become markedly more practical with the advent of the fused-silica capillary column, computer-controlled chromatography and data analysis, simplified sample preparation, and a commercially available GLC system dedicated to microbiological applications. Experience with applications in diagnostic microbiology ranges from substantial success in work with mycobacteria, legionellae, and nonfermentative gram-negative bacilli to minimal involvement with fungi and other nonbacterial agents. GLC is a good alternative to other means for the identification of mycobacteria or legionellae because it is rapid, specific, and independent of other specialized testing, e.g., DNA hybridization. Nonfermenters show features in their cellular fatty acid content that are useful in identifying species and, in some cases, subspecies. Less frequently encountered nonfermenters, including those belonging to unclassified groups, can ideally be characterized by GLC. Information is just beginning to materialize on the usefulness of cellular fatty acids for the identification of gram-positive bacteria and anaerobes, despite the traditional role of GLC in detecting metabolic products as an aid to identification of anaerobes. When species identification of coagulase-negative staphylococci is called for, GLC may offer an alternative to biochemical testing. Methods for direct analysis of clinical material have been developed, but in practical and economic terms they are not yet ready for use in the clinical laboratory. Direct analysis holds promise for detecting markers of infection due to an uncultivable agent or in clinical specimens that presently require cultures and prolonged incubation to yield an etiologic agent.


A critical review on lithium-air battery electrolytes.
Metal-air batteries, utilizing the reduction of ambient oxygen, have the highest energy density because most of the cell volume is occupied by the anode while the cathode active material is not stored in the battery. Lithium metal is a tempting anode material for any battery because of its outstanding specific capacity (3842 mA h g(-1) for Li vs. 815 mA h g(-1) for Zn). Combining the high energy density of Li with ambient oxygen seems to be a promising option. Specifically, in all classes of electrolytes, the transformation from Li-O2 to Li-air is still a major challenge as the presence of moisture and CO2 reduces significantly the cell performance due to their strong reaction with Li metal. Thus, the quest for electrolyte systems capable of providing a solution to the imposed challenges due to the use of metallic Li, exposure to the environment and handling the formation of reactive discharged product is still on. This extended Review provides an expanded insight into electrolytes being suggested and researched and also a future vision on challenges and their possible solutions.


Epidemiology, control and prevention of tobacco induced oral mucosal lesions in India.
Oral cancer is one of the leading causes of human morbidity and mortality especially in developing countries like India. Tobacco consumption in smokeless and smoking form along with alcohol is considered as the primary risk factors. Tobacco is a major health challenge with various tobacco products available for use which are known to have deleterious effects on the oral mucosa. The oral lesions caused by tobacco are inclusive of those that are less likely to progress to cancer; lesions with increased tendency to develop into cancer and cancerous lesions. Prevention and control of tobacco induced oral mucosal lesions is the prime requisite currently and mainly involves measures undertaken at primary, secondary and tertiary levels. Primary prevention plays a pivotal role in tobacco induced lesions and steps can be taken at policy level, community as well as individual level. This review paper focuses on the epidemiological data of tobacco induced oral mucosal lesions in India available in the literature with an overview on various strategies for their prevention and control.


Progression through mitosis promotes PARP inhibitor-induced cytotoxicity in homologous recombination-deficient cancer cells.
Mutations in homologous recombination (HR) genes BRCA1 and BRCA2 predispose to tumorigenesis. HR-deficient cancers are hypersensitive to Poly (ADP ribose)-polymerase (PARP) inhibitors, but can acquire resistance and relapse. Mechanistic understanding how PARP inhibition induces cytotoxicity in HR-deficient cancer cells is incomplete. Here we find PARP inhibition to compromise replication fork stability in HR-deficient cancer cells, leading to mitotic DNA damage and consequent chromatin bridges and lagging chromosomes in anaphase, frequently leading to cytokinesis failure, multinucleation and cell death. PARP-inhibitor-induced multinucleated cells fail clonogenic outgrowth, and high percentages of multinucleated cells are found in vivo in remnants of PARP inhibitor-treated Brca2-/-;p53-/- and Brca1-/-;p53-/- mammary mouse tumours, suggesting that mitotic progression promotes PARP-inhibitor-induced cell death. Indeed, enforced mitotic bypass through EMI1 depletion abrogates PARP-inhibitor-induced cytotoxicity. These findings provide insight into the cytotoxic effects of PARP inhibition, and point at combination therapies to potentiate PARP inhibitor treatment of HR-deficient tumours.


Genetic diversity and relationship of chicory (Cichorium intybus L.) using sequence-related amplified polymorphism markers.
Chicory is a crop with economically important roles and is cultivated worldwide. The genetic diversity and relationship of 80 accessions of chicories and endives were evaluated by sequence-related amplified polymorphism (SRAP) markers to provide a theoretical basis for future breeding programs in China. The polymorphic rate was 96.83%, and the average polymorphic information content was 0.323, suggesting the rich genetic diversity of chicory. The genetic diversity degree of chicory was higher (GS = 0.677) than that of endive (GS = 0.701). The accessions with the highest genetic diversity (effective number of alleles, NE = 1.609; Nei's genetic diversity, H = 0.372; Shannon information index, I = 0.556) were from Italy. The richest genetic diversity was revealed in a chicory line (NE = 1.478, H = 0.289, I = 0.443) among the 3 types (line, wild, and cultivar). The chicory genetic structure of 8 geographical groups showed that the genetic differentiation coefficient (GST) was 14.20% and the number of immigrants per generation (Nm) was 3.020. A GST of 6.80% and an Nm of 6.853 were obtained from different types. This observation suggests that these chicory lines, especially those from the Mediterranean region, have potential for providing rich genetic resources for further breeding programs, that the chicory genetic structure among different countries obviously differs with a certain amount of gene flow, and that SRAP markers could be applied to analyze genetic relationships and classifications of Cichorium intybus and C. endivia.


Prophylaxis against pulmonary embolism in patients undergoing total hip arthroplasty.
The optimal method of prophylaxis for the prevention of pulmonary embolism in patients undergoing total hip arthroplasty remains controversial. Guidelines appear to be contradictory. The purpose of the present study was to examine whether a best prophylactic agent exists for the prevention of postoperative pulmonary embolism and whether the type of anesthesia affects the rates of pulmonary embolism. From 2001 to 2008, a total joint registry from a nationwide health maintenance organization was evaluated to determine the rates of pulmonary embolism, fatal pulmonary embolism, and death among 17,595 patients without a history of venous thromboembolism who were managed with unilateral total hip arthroplasty. All patients were followed for ninety days postoperatively. Data were abstracted electronically and were validated through chart reviews. Multivariate logistic regression models were used to assess associations between the types of prophylaxis and anesthesia that were used and pulmonary embolism while adjusting for other risk factors. Patients received either mechanical prophylaxis alone (N = 1533) or chemical prophylaxis (aspirin [N = 934], Coumadin [warfarin] [N = 6063], or low-molecular-weight heparin [N = 7202]) with or without mechanical prophylaxis. The rate of pulmonary embolism was 0.41% (95% confidence interval [CI], 0.32% to 0.51%) overall, 0.37% (95% CI, 0.05% to 0.70%) for mechanical prophylaxis, 0.43% (95% CI, 0.01% to 0.85%) for aspirin, 0.43% (95% CI, 0.26% to 0.59%) for Coumadin, 0.40% (95% CI, 0.26% to 0.55%) for low-molecular-weight heparin, 0.43% (95% CI, 0.28% to 0.58%) for general anesthesia, and 0.40% (95% CI, 0.28% to 0.52%) for non-general anesthesia. The mortality rate was 0.51% (95% CI, 0.40% to 1.01%) overall, 0.67% (95% CI, 0.23% to 1.34%) for mechanical prophylaxis, 0.64% (95% CI, 0.13% to 1.28%) for aspirin, 0.51% (95% CI, 0.33% to 1.02%) for Coumadin, 0.42% (95% CI, 0.27% to 0.83%) for low-molecular-weight heparin, 0.51% (95% CI, 0.35% to 0.67%) for general anesthesia, and 0.50% (95% CI, 0.36% to 0.64%) for non-general anesthesia. Regression models did not show any association between the type of prophylaxis used or the choice of anesthesia and increased odds of pulmonary embolism when adjusting for age, sex, and American Society of Anesthesiologists score. No clinical differences were detected among the types of prophylaxis against venous thromboembolism or the types of anesthesia with respect to pulmonary embolism, fatal pulmonary embolism, or death on the basis of prospective collection of data by a contemporary total joint registry.


Identification of tumor-intrinsic drivers of immune exclusion in acral melanoma.
Acral melanoma (AM) has distinct characteristics as compared with cutaneous melanoma and exhibits poor response to immune checkpoint inhibitors (ICIs). Tumor-intrinsic mechanisms of immune exclusion have been identified in many cancers but less studied in AM. We characterized clinically annotated tumors from patients diagnosed with AM at our institution in correlation with ICI response using whole transcriptome RNAseq, whole exome sequencing, CD8 immunohistochemistry, and multispectral immunofluorescence imaging. A defined interferon-γ-associated T cell-inflamed gene signature was used to categorize tumors into non-T cell-inflamed and T cell-inflamed phenotypes. In combination with AM tumors from two published studies, we systematically assessed the immune landscape of AM and detected differential gene expression and pathway activation in a non-T cell-inflamed tumor microenvironment (TME). Two single-cell(sc) RNAseq AM cohorts and 11 bulk RNAseq cohorts of various tumor types were used for independent validation on pathways associated with lack of ICI response. In total, 892 specimens were included in this study. 72.5% of AM tumors showed low expression of the T cell-inflamed gene signature, with 23.9% of total tumors categorized as the non-T cell-inflamed phenotype. Patients of low CD3+CD8+PD1+ intratumoral T cell density showed poor prognosis. We identified 11 oncogenic pathways significantly upregulated in non-T cell-inflamed relative to T cell-inflamed TME shared across all three acral cohorts (MYC, HGF, MITF, VEGF, EGFR, SP1, ERBB2, TFEB, SREBF1, SOX2, and CCND1). scRNAseq analysis revealed that tumor cell-expressing pathway scores were significantly higher in low versus high T cell-infiltrated AM tumors. We further demonstrated that the 11 pathways were enriched in ICI non-responders compared with responders across cancers, including AM, cutaneous melanoma, triple-negative breast cancer, and non-small cell lung cancer. Pathway activation was associated with low expression of interferon stimulated genes, suggesting suppression of antigen presentation. Across the 11 pathways, fatty acid synthase and CXCL8 were unifying downstream target molecules suggesting potential nodes for therapeutic intervention. A unique set of pathways is associated with immune exclusion and ICI resistance in AM. These data may inform immunotherapy combinations for immediate clinical translation.


Combat readiness for the modern military surgeon: data from a decade of combat operations.
Hundreds of general surgeons from the army, navy, and air force have been deployed during the past 10 years to support combat forces, but little data exist on their preparedness to handle the challenging injuries that they are currently encountering. Our objective was to assess operative and operational experience in theater with the goal of improving combat readiness among surgeons. A detailed survey was sent to 246 active duty surgeons from the army, navy, and air force who have been deployed at least once in the past 10 years, requesting information on cases performed, perceptions of efficacy of predeployment training, knowledge deficits, and postdeployment emotional challenges. Survey data were kept confidential and analyzed using standard statistical methods. Of 246 individuals, 137 (56%) responded and 93 (68%) have been deployed two or more times. More than 18,500 operative procedures were reported, with abdominal and soft tissue cases predominating. Many surgeons identified knowledge or practice gaps in predeployment vascular (46%), neurosurgical (29.9%), and orthopedic (28.5%) training. The personal burden of deployment manifested itself with both family (approximately 10% deployment-related divorce rate) and personal (37 surgeons [27%] with two or more symptoms of posttraumatic stress syndrome) stressors. These data support modifications of predeployment combat surgical training to include increased exposure to open vascular procedures and curriculum traditionally outside general surgery (neurosurgery and orthopedics). The acute care surgical model may be ideal for the military surgeon preparing for deployment. Further research should be directed toward identifying factors contributing to psychological stress among military medics.


Joint Attention and Sensory-Regulatory Features at 13 and 22 Months as Predictors of Preschool Language and Social-Communication Outcomes.
Purpose The purpose of this study was to understand how joint attention and sensory-regulatory features are related in early childhood and predict language and social-communication outcomes in preschool in order to build mechanistic theories that can inform early intervention directed at improving these outcomes. Method Cross-lagged panel analysis models were used to examine the association between joint attention and sensory-regulatory features at 13 and 22 months of age in children (n = 87) who were identified via community screening at 12 months as having a higher likelihood than the general population for being diagnosed with autism spectrum disorder. Results Significant concurrent correlations and predictive correlations were found between these constructs at 13 and 22 months. Joint attention skills at 13 months predicted both joint attention and sensory-regulatory features at 22 months. Distal language and social-communication outcomes at preschool age (n = 48) were best predicted by sensory-regulatory features at 22 months. Conclusions Both joint attention and sensory regulation are important factors in the first and second years of life for impacting later preschool language and social-communication outcomes in this sample. These findings may have implications for future early childhood intervention research for children at a higher likelihood for autism spectrum disorder.


[Depakin 300 and Depakin-chrono in the therapy of epilepsy].
The results of administration of Depakin 300 and Depakin-chrono during 4-year period were analysed in 137 patients aged 14-36 years with idiopathic generalized, cryptogenic, symptomatic partial and nonclassified epilepsy. Depakin was prescribed as basic medicine in idiopathic generalized epilepsy, while in partial and nonclassified epilepsy this drug was more frequently administered in combination with some other anticonvulsants. Early administration of Depakin was highly effective: a remission was observed in 91.2% of the patients during a year. During 3 years a remission was observed in 75.4%, frequency of the fits decreased more than by 50%--in 14.1% of the cases, no response was found in 10.5% of the patients (in cases of gross structural changes of the brain, inefficiency of the preceding therapy during 5 years). Frequency of side-effects was 1.7-12.3%. Indications to treatment are discussed: in addition to generally accepted recommendations (absence, myoclonic primary generalized, tonic-clonic forms of epilepsy, of reading, photosensitive epilepsy), application of Depakin is also recommended in nonclassified epilepsy with generalized epileptic EEG-patterns as well as a drug of choice in all cases of nonclassified fits. Depakin is also quite efficient as an additional drug in cases of partial epilepsy resistant to treatment.


The development of novel organically modified sol-gel media for use with ATR/FTIR sensing.
The ability to prepare and develop novel pre-concentration media by the sol-gel process, and their integration with mid-infrared transparent waveguides has been demonstrated. This research approach resulted in a mid-infrared sensing methodology in which the properties (porosity, functionality, polarity, etc.) of the recognition layer could be tailored by variation of the sol-gel precursors and processing conditions. Cross-linker type and concentration notably influenced p-xylene absorption and diffusion rate. Unreacted silanol groups appeared to be the dominant factor in the hydrophobicity of sol-gel layers. Variation of sol-gel precursors and thermal treatment altered both film cross-link density and polarity, as demonstrated by variation in the rate of analyte diffusion and equilibrium analyte concentration. The use of a novel 1 : 1 PTMOS : DPDMS material as pre-concentration medium in this analytical sensing approach was validated through the determination of p-nitrochlorobenzene in an aqueous environment. The response demonstrated linearity between 0-30 mg L(-1) with a correlation coefficient of 0.989 and a limit of detection of 0.7 mg L(-1). Sensing times for p-nitrochlorobenzene were also reduced from several hours to 24 minutes, without loss of measurement accuracy or sensitivity, by a 10 degrees C increase in the sensing temperature and the use of a predictive Fickian model previously developed by this research group.


Gender differences in detecting unanticipated stimuli: an ERP study.
The aim of the present study was to investigate the gender differences during an emotional anticipation task. Sixteen females and sixteen males participated in the experiment. Event-related potentials (ERPs) were measured in a modified cue-target paradigm and were recorded following stimuli that differed in two dimensions: (1) predictable vs. unpredictable pictures and (2) negative vs. neutral pictures. Cue-induced ERP results demonstrated that females had enhanced positive component (P2) compared to males. Moreover, results showed that during the unpredictable condition, females displayed larger P2 amplitudes in negative and neutral anticipation than males. This study demonstrates that females have greater sensitivity to the unanticipated stimuli, which may contribute to evolution.


Impact of Crossing Strategy on Intermediate-term Outcomes After Chronic Total Occlusion Percutaneous Coronary Intervention.
There is ongoing controversy about the optimal crossing strategy selection for chronic total occlusion (CTO) percutaneous coronary intervention (PCI), especially regarding the relative merits of antegrade dissection/re-entry and the retrograde approach. We retrospectively examined the clinical outcomes of 173 consecutive patients who underwent successful CTO PCI at our institution between January 2012 and March 2015. The mean age was 65 ± 8 years, and 98% of the patients were men with a high prevalence of diabetes (60%), previous coronary artery bypass grafting (CABG) (31%), and previous PCI (54%). The successful CTO crossing strategy was antegrade wire escalation in 79 patients (45.5%), antegrade dissection/re-entry in 58 patients (33.5%), retrograde wire escalation in 11 patients (6.4%), and retrograde dissection and re-entry in 25 patients (14.5%). The retrograde approach was more commonly used in lesions with interventional collaterals (P < 0.0001), moderate/severe calcification (P = 0.02), blunt stump (P = 0.01), and a higher Japan Chronic Total Occlusion score (P = 0.0002). Use of dissection and re-entry (both antegrade and retrograde) was associated with bifurcation and the distal cap (P = 0.004), longer CTO occlusion length (P < 0.0001), and longer stent length (P < 0.0001). Median follow-up was 11 months. The 12-month incidence of death, myocardial infarction, and the composite of acute coronary syndrome/target lesion revascularization/target vessel revascularization was 2.5%, 4.9%, and 24.4%, respectively, and was similar with intimal and subintimal crossing strategies. Antegrade dissection/re-entry and retrograde approaches are frequently used during CTO PCI and were associated with similarly favorable intermediate-term outcomes as antegrade wire escalation.


MLST typing of Treponema pallidum subsp. pallidum in the Czech Republic during 2004-2017: Clinical isolates belonged to 25 allelic profiles and harbored 8 novel allelic variants.
A recently introduced Multilocus Sequence Typing scheme for Treponema pallidum subsp. pallidum was applied to clinical samples collected from 2004 to 2017 from the two largest cities (Prague and Brno) in the Czech Republic. Altogether, a total of 675 samples were tested in this study and 281 of them were found PCR-positive for treponemal DNA and typeable. Most of the typed samples (n = 281) were swabs from primary or secondary syphilis lesions (n = 231), and only a minority were whole blood or tissue samples (n = 50). Swab samples from patients with rapid plasma regain (RPR) values of 1-1024 were more frequently PCR-positive (84.6%) compared to samples from patients with non-reactive RPR test (46.5%; p-value = 0.0001). Out of 281 typeable samples, 136 were fully-typed at all TP0136, TP0548, and TP0705 loci. Among the fully and partially typed samples, 25 different allelic profiles were identified. Altogether, eight novel allelic variants were found among fully (n = 5) and partially (n = 3) typed samples. The distribution of TPA allelic profiles identified in the Czech Republic from 2004 to 2017 revealed a dynamic character with allelic profiles disappearing and emerging over time. While the number of samples with the A2058G mutation was seen to increase (86.7% in 2016/2017), the number of samples harboring the A2059G mutation was found to have decreased over time (3.3% in 2016/2017). In addition, we found several allelic profile associations with macrolide resistance or susceptibility, the gender of patients, as well as patient residence.


[Nitrogen balance assessment in burn patients].
The burn injury probably represents the largest stimulus for muscle protein catabolism. This state is characterized by an accelerated catabolism of the lean or skeletal mass that results in a clinical negative balance of nitrogen and muscle wasting. The determination of an appropriate value for protein intake is essential, since it is positively related to the nitrogen balance (NB) and accordingly several authors argue that a positive NB is the key parameter associated with nutritional improvement of a burn patient. Evaluation of the degree of protein catabolism by assessment of the Nitrogen Balance; Defining of nutritional support (protein needs) to implement in patients with burned surface area (BSA) = 10%. We prospectively evaluated the clinical files and scrutinized the clinical variables of interest. The NB was estimated according to three formulae. Each gram of nitrogen calculated by the NB was then converted into grams of protein, subtracted or added to protein intake (or administered enteric or parenterically) and divided by kg of reference Weight (kg Rweight), in an attempt to estimate the daily protein needs. The cohort consisted of 10 patients, 6 females, with average age of 58(23) years old, a mean of BSA of 21.4(8.4)%, ranging from a minimum of 10.0% and máximum of 35.0%. On average, patients were 58 (23) years old. The average number of days of hospitalization in the burn unit was 64.8(36.5) days. We observed significant differences between the 3 methods used for calculating the NB (p = 0.004), on average the NB was positive. When the formula A was used the average value of NB was higher. Regarding the attempt to estimate the needs of g prot/kg Rweight/day most of the values did not exceed, on average, 2.6 g Prot/kg Rweight/day and no significant differences between patients with a BSA% of 10-20% and with BSA% > 20% were found. Despite being able to estimate the protein catabolism through these formulas and verifying that most values were above zero, wide individual fluctuations were visible over time. Based on the sample reference that recommends a value of 1.5-2 g Prot/kg Rweight/day, we can conclude it to be underestimated, when comparing with the mean value of 2.6 g Prot/kg Rweight/day we established.


Lightning Rods, Earthquakes, and Regional Identities: Towards a Multi-Scale Framework of Assessing Fracking Risk Perception.
Hydraulic fracturing has provided a persistent, polarizing, and highly politicized source of controversy internationally and in numerous national contexts for just under a decade. This research uses hydraulic fracturing (i.e., fracking) operations in New Zealand as a vignette through which to understand the underlying causes of controversy and the appropriateness of attempts to address them. A multi-method approach using interviews (n = 25), diagrammatic analysis, and newsprint media was applied to evidence two major findings. First, previous attempts to explain fracking controversy based on social constructivist theory lack a multi-scalar approach to the assessment of factors that influence risk perceptions. It is found that risk perception surrounding fracking in New Zealand reflects intra-scalar interactions between factors originating at the international, national, regional, and local scale. Second, there is a concerning absence of critique pertaining to the concept of "social license to operate" (SLO), which has been advocated both internationally and nationally as an appropriate form of stakeholder engagement. This article contributes to the SLO outcomes literature by establishing a need to consider multi-scalar influences on risk perception when explaining diverse SLO outcomes in communities where fracking operations are prospective or already taking place.


[Perceptions and use of Universal Health Coverage (Plan Sésame) among the elderly in Dakar (Senegal), impacts on health expenditure related to diabetes and hypertension].
In 2006, the Senegalese government set up a health coverage programme for people aged 60 and over - the Plan Sésame - to provide free medical care in all the country's public health facilities. This scheme has been integrated into the Universal Health Coverage (CMU) promoted from 2013. The objective of the study was to describe and analyse the knowledge and representations of professionals and users about health coverage and the Plan Sésame, the use of the scheme by the elderly, to evaluate the amount of medical expenses incurred during a routine medical consultation for the monitoring of their illness (hypertension and diabetes), and to calculate the out-of-pocket expenses related to the consultation. Study conducted between July 2020 and October 2021 in two public health facilities in Dakar. Mixed approach: 1/ qualitative study by semi-directive interviews, informal interviews, observations and field diary with 35 people selected according to a reasoned choice procedure with the aim of diversifying gender, age, social status, therapeutic itineraries for 23 people (including 12 women, ages between 60 and 85 years), and professional activities for 12 health actors; 2/ quantitative cross-sectional study by questionnaire of 225 people (including 141 women) aged 60 and over; we calculated the total cost of the consultation and associated prescriptions (complementary examinations and medicines) as well as the remaining medical expenses (out-of-pocket) and the cost of transporting patients. This is a descriptive exploratory study of a non-representative sample of the elderly population in Senegal. The health professionals interviewed supported the principle of health coverage, but most of them had limited and sometimes imprecise knowledge of the existing schemes and the methods of access or the services covered. Their point of view about the consequences of the Plan Sésame on their practice reveals some contradictions: some complain about the increase in workload, the criticism is extended to all the free schemes which would have a negative impact on daily practice because of the increase in the number of consultations which would be linked to abuse by patients.The interviews highlight the heterogeneity of the knowledge of elderly people about the health coverage intended for them, even though the Plan Sésame has been in place for over ten years. The interviews clearly show that the use of the health coverage system by the elderly depends closely on the information they have and their ability to use it, both for women and men. There is a close link between the level of social integration of people and their use of health coverage: the most socially integrated people are those who know how to use CMU services best. The use of health coverage by the elderly appears to vary according to the individual.Although Plan Sésame is defined as part of a national strategy, its implementation varies according to the health structures and the periods; in the two study sites, the range of services covered by Plan Sésame is very limited, so the coverage provided by Plan Sésame is only partial: between 30 and 50% of the medical costs; the remaining cost of a consultation for elderly patients with hypertension and/or diabetes varies between 24,000 and 28,000 CFA francs.These amounts must be put into perspective with the resources available to people. Statistical studies published in 2021 report that in Senegal the average daily expenditure is 1,390 CFA francs/person/day; and that almost 38% of the population lives on 913 CFA francs/person/ day, which is the poverty line calculated in 2019. Thus, the average out-of-pocket expenses for a follow-up consultation for hypertension, diabetes or a combination of the two diseases represent 15 to 30 days of daily expenditure. While the vast majority of elderly people in Senegal do not have a retirement pension, health expenses are therefore borne by their relatives. Within households, medical expenditure for the elderly competes with basic needs, particularly food, which usually take up more than half of household resources. This indispensable family support places the elderly in a situation of total dependence. In 2021, Plan Sésame does not yet allow for completely free care for the elderly. However, its application, even partial, has resulted in a real reduction in health care costs for the elderly. Its use remains limited due to inconsistent application by most health structures. Its impact is insufficient in view of the amounts that users have to pay in a context of social and economic vulnerability. These observations reinforce the need to work on reducing the price of medical services and strengthening the UHC, in order to improve the equity and performance of the system, and to make it fully functional in all health structures.


Plasma inflammatory biomarkers link to diffusion tensor imaging metrics in virally suppressed HIV-infected individuals.
Inflammation may contribute to brain white matter health in people living with HIV who report cognitive symptoms despite adherence to combination antiretroviral therapy and viral suppression. We explored relationships between diffusion tensor imaging (DTI) metrics of white matter, plasma biomarkers of immune activation, and cognitive function in the HIV-infected population. Retrospective study of older adults living with HIV who are combination antiretroviral therapy adherent, virally suppressed, and self-report cognitive symptoms. MRI, blood draws, and standardized neuropsychological test scores were collected from HIV-infected individuals. DTI metrics (fractional anisotropy, mean diffusivity, radial diffusivity, axial diffusivity) and plasma biomarkers (soluble CD163, soluble CD14, neopterin, IFN γ-induced protein 10, monocyte chemoattractant protein 1) were quantified. Statistical analysis explored associations between biomarker levels or neuropsychological test scores and DTI metrics using region of interest analyses and a voxelwise approach. A total of 43 participants with median (interquartile range) age of 64 (62-66 years), CD4 cell count of 600 (400-760 cell/μl) who were all virally suppressed (<100 copies/ml) were selected. Higher levels of monocyte chemoattractant protein 1 associated with lower fractional anisotropy and higher mean diffusivity (P < 0.05) across white matter tracts including corpus callosum, corona radiata, and superior longitudinal fasciculus. Higher neopterin associated with higher mean diffusivity in the genu of corpus callosum, and higher soluble CD14 associated with lower fractional anisotropy in the bilateral superior corona radiata (P < 0.05). Worse global performance and speed domain scores associated with higher mean diffusivity and lower fractional anisotropy, and worse executive domain scores associated with lower fractional anisotropy (P < 0.05). Elevated inflammatory plasma biomarkers link to white matter abnormalities among virally suppressed individuals. DTI abnormalities associate to cognitive performance. We conclude that inflammatory processes impact clinically relevant brain health indices despite viral suppression.


Human JIK, a novel member of the STE20 kinase family that inhibits JNK and is negatively regulated by epidermal growth factor.
Mammalian members related to Saccharomyces cerevisiae serine/threonine kinase STE20 can be divided into two subfamilies based on their structure and function. The PAK subfamily is characterized by an N-terminal p21-binding domain (also known as CRIB domain), a C-terminal kinase domain, and is regulated by the small GTP-binding proteins Rac1 and Cdc42Hs. The second group is represented by the GCK-like members, which contain an N-terminal catalytic domain and lack the p21-binding domain. Some of them have been demonstrated to induce c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) cascade, while others have been shown to be activated by a subset of stress conditions or apoptotic agents, although little is known about their specific function. Here, we have identified a novel human STE20-related serine/threonine kinase, belonging to the GCK-like subfamily. This kinase does not induce the JNK/SAPK pathway, but, instead, inhibits the basal activity of JNK/SAPK, and diminishes its activation in response to human epidermal growth factor (EGF). Therefore, we designated this molecule JIK for JNK/SAPK-inhibitory kinase. The inhibition of JNK/SAPK signaling pathway by JIK was found to occur between the EGF receptor and the small GTP-binding proteins Rac1 and Cdc42Hs. In contrast, JIK does not activate nor does it inhibit ERK2, ERK6, p38, or ERK5. Furthermore, JIK kinase activity is not modulated by any exogenous stimuli, but, interestingly, it is dramatically decreased upon EGF receptor activation. Thus, JIK might represent the first member of the STE20 kinase family whose activity can be negatively regulated by tyrosine kinase receptors, and whose downstream targets inhibit, rather than enhance, JNK/SAPK activation.


Penetration injury to the heart. Nine consecutive cases at a community hospital.
During the past 25 years, great progress has been made in saving patients with serious cardiac injuries. This study is based on nine consecutive cases of penetrating wounds to the heart treated successfully by surgical intervention at a community hospital with no facilities for cardiopulmonary bypass. Early surgery is advocated for patients suspected of having penetrating injury of the heart.


Pleural effusion due to Parvimonas micra. A case report and a literature review of 30 cases.
The clinical and microbiological characteristics of infections caused by Parvimonas micra is described, including 30 cases in the literature and a new case handled at the present centre. Out of the 31 patients, 18 were male; mean age at diagnosis was 65.1 ± 13.0 years. Infection site was the vertebral spine in 14 patients and joints and heart valves in 5 each one; pain was present in all patients with articular localization and in almost all patients with vertebral involvement. The diagnosis was obtained from fluid aspirate or drainage in 13 cases and blood cultures in 11. In 8 cases, molecular techniques were also applied. The most frequently used antimicrobials were clindamycin, penicillin, amoxicillin and ceftriaxone. The outcome was positive with the medical treatment in 28 patients. P. micra infections are uncommon and requires a high index of suspicion.


Genome sequencing and comparative analysis of three Chlamydia pecorum strains associated with different pathogenic outcomes.
Chlamydia pecorum is the causative agent of a number of acute diseases, but most often causes persistent, subclinical infection in ruminants, swine and birds. In this study, the genome sequences of three C. pecorum strains isolated from the faeces of a sheep with inapparent enteric infection (strain W73), from the synovial fluid of a sheep with polyarthritis (strain P787) and from a cervical swab taken from a cow with metritis (strain PV3056/3) were determined using Illumina/Solexa and Roche 454 genome sequencing. Gene order and synteny was almost identical between C. pecorum strains and C. psittaci. Differences between C. pecorum and other chlamydiae occurred at a number of loci, including the plasticity zone, which contained a MAC/perforin domain protein, two copies of a >3400 amino acid putative cytotoxin gene and four (PV3056/3) or five (P787 and W73) genes encoding phospholipase D. Chlamydia pecorum contains an almost intact tryptophan biosynthesis operon encoding trpABCDFR and has the ability to sequester kynurenine from its host, however it lacks the genes folA, folKP and folB required for folate metabolism found in other chlamydiae. A total of 15 polymorphic membrane proteins were identified, belonging to six pmp families. Strains possess an intact type III secretion system composed of 18 structural genes and accessory proteins, however a number of putative inc effector proteins widely distributed in chlamydiae are absent from C. pecorum. Two genes encoding the hypothetical protein ORF663 and IncA contain variable numbers of repeat sequences that could be associated with persistence of infection. Genome sequencing of three C. pecorum strains, originating from animals with different disease manifestations, has identified differences in ORF663 and pseudogene content between strains and has identified genes and metabolic traits that may influence intracellular survival, pathogenicity and evasion of the host immune system.


Trust Building Recruitment Strategies for Researchers Conducting Studies in African American (AA) Churches: Lessons Learned.
An initial and vital important step in recruiting participants for church-based hospice and palliative care research is the establishment of trust and credibility within the church community. Mistrust of medical research is an extremely important barrier hindering recruitment in African American (AA) communities. A church-based EOL dementia education project is currently being conducted at four large urban AA churches. Church leaders voiced mistrust concerns of previous researchers who conducted investigations in their faith-based institutions. We explored strategies to ameliorate the mistrust concerns. Specific aim: To identify trust-rebuilding elements for researchers following others who violated trust of AA church leaders. Face-to-face, in-depth interviews were conducted from a convenient sample of four established AA church leaders. Interviews were held in the informants' churches to promote candor and comfort in revealing sensitive information about trust /mistrust. Content analysis framework was used to analyze the data. Elements identified from the analysis were then used to create themes. Multidimensional overarching themes emerged from the analysis included: Experience with researchers (positive and extremely negative), violation of trust and trust building strategies. Findings suggest that researchers who wish to conduct successful studies in the AA religious institutions must implement trust rebuilding strategies that include mutual respect, collaboration and partnership building. If general moral practices continue to be violated, threat to future hospice and palliative care research within the institutions may prevail. Thus, potential benefits are thwarted for the church members, AA community, and advancement of EOL care scholarship.


[Present-day aspects (diagnosis, clinical course and treatment) of acute progressive pulmonary tuberculosis].
A total of 103 patients with acute progressive pulmonary tuberculosis whose age ranged from 18 to 60 years were examined. Caseous pneumonic, infiltrative-caseous, disseminated, and rapidly progressive fibrocavernous tuberculosis was found in 45.6, 17.5, 16.5, and 20.4% of cases, respectively. The clinical picture was characterized by its acute onset with significant intoxication syndrome. Moreover, all the patients had respiratory and immunological failure, varying disseminated intravascular coagulation syndrome, non-specific bronchopulmonary infection; some presented with pulmonary hemorrhage, spontaneous pneumothorax and pleural empyema. The sputum smear test was positive in all the patients. If there was no evidence for drug resistance, patients had a 4-month regimen using isoniazid, rifampicin, pyrazinamide, ethembutol, and kanamycin. In the subsequent 10-12 months, isoniazid, rifampicin, and ethambutol were given. The patients with multidrug resistant tuberculosis were administered protionamide, ofloxacin, amikacin, supplemented by pyrazinamide and ethambutol. The combined chemotherapy could stop bacterial isolation in more than 80% of patients, make the process stable, and prepare them for planned surgical treatment. When complications occurred and the disease was in steady progress, salvage operations were made, which was the only possible way of preventing the progression of disease at times and of saving life in the patient.


Quantification of hydrogen production by intestinal bacteria that are specifically dysregulated in Parkinson's disease.
Oral administration of hydrogen water ameliorates Parkinson's disease (PD) in rats, mice, and humans. We previously reported that the number of putative hydrogen-producing bacteria in intestinal microbiota is low in PD compared to controls. We also reported that the amount of hydrogen produced by ingestion of lactulose is low in PD patients. The decreased hydrogen production by intestinal microbiota may be associated with the development and progression of PD. We measured the amount of hydrogen production using gas chromatography by seven bacterial strains, which represented seven major intestinal bacterial groups/genera/species. Blautia coccoides and Clostridium leptum produced the largest amount of hydrogen. Escherichia coli and Bacteroides fragilis constituted the second group that produced hydrogen 34- to 93-fold lower than B. coccoides. Bifidobacterium pseudocatenulatum and Atopobium parvulum constituted the third group that produced hydrogen 559- to 2164-fold lower than B. coccoides. Lactobacillus casei produced no detectable hydrogen. Assuming that taxonomically neighboring strains have similar hydrogen production, we simulated hydrogen production using intestinal microbiota that we previously reported, and found that PD patients produce a 2.2-fold lower amount of intestinal hydrogen compared to controls. The lower amount of intestinal hydrogen production in PD was also simulated in cohorts of two other countries. The number of hydrogen-producing intestinal bacteria may be associated with the development and progression of PD. Further studies are required to prove its beneficial effect.


In vivo modulation and quantification of microRNAs during axolotl tail regeneration.
The ability to regenerate diseased, injured, or missing complex tissue is widespread throughout lower vertebrates and invertebrates; however, our knowledge of the molecular mechanisms that regulate this amazing ability is still in its infancy. Many recent papers have shown important roles for microRNAs in regulating regeneration in a number of species. The ability to detect and quantify miRNA expression fluctuations at a single cell level in vivo in different cell types during processes like regeneration is very informative. In this chapter, we describe how to use a dual-fluorescent green fluorescent protein (GFP)-reporter/monomeric red fluorescent protein (mRFP)-sensor (DFRS) plasmid to quantitate the dynamics of specific miRNAs over time following miRNA mimic injection as well as during regeneration. In this bicistronic vector, the mRFP allows for verification of miRNA expression, while the GFP functions as an internal control to normalize miRNA expression and thus obtain quantitative results. In addition, we demonstrate how this technique revealed dynamic miR-23a expression and function during tail regeneration.


Regression to Normal Glucose Regulation in American Indians and Alaska Natives of a Diabetes Prevention Program.
This study evaluated whether regression from impaired glucose regulation (IGR) to normal glucose regulation (NGR) after 1 year of a lifestyle intervention reduces diabetes risk in American Indians and Alaska Natives (AI/ANs). In addition, we sought to identify predictors for regression to NGR and understand possible mechanisms for the association between NGR and future diabetes risk. Data from participants enrolled from 2006 to 2009 in the Special Diabetes Program for Indians Diabetes Prevention Program with IGR at baseline and an oral glucose tolerance test at year 1 were analyzed (N = 1,443). Cox regression models were used to estimate the subsequent diabetes risk (year 1 to year 3) by year 1 glucose status. Mediation analysis was used to estimate the proportions of the association between year 1 glycemic status and diabetes risk explained by specific factors. Those who reverted to NGR at year 1 (38%) had lower diabetes risk than those with sustained IGR (adjusted hazard ratio 0.28, 95% CI 0.12-0.67). The lower risk associated with regression to NGR was explained by both baseline risk factors and differences in weight loss. Metformin use, weight loss, and an increase in exercise were modifiable risk factors associated with higher odds of regression to NGR. Patients with prediabetes who reverted to NGR had a reduced risk of developing type 2 diabetes over the next 2 years. Both baseline and modifiable risk factors explained the risk reduction associated with NGR.


B. F. Skinner and technology's nation: Technocracy, social engineering, and the good life in 20th-century America.
Psychologist B. F. Skinner developed and promoted a technology of behavior as the basis for widespread social reform over much of his career. In 1948, he published his behaviorally engineered vision of the good life in his utopian novel Walden Two (Skinner, 1948). Skinner's efforts were part of a much larger social engineering tradition that received one of its fullest expressions in the Technocracy Movement of the 1930s. Fifteen years before Skinner's Walden Two, at the height of the Technocracy Movement's public visibility in the United States, technocrat Harold Loeb (1933/1996) published his utopia, Life in a Technocracy: What It Might Be Like. In this article, I place the socially engineered visions of the good life promoted by the Technocracy Movement and by Skinner on an intellectual and ideological continuum to amplify and explore American attitudes toward psychology, technology, and social engineering during the middle decades of the 20th century. I argue that responses to both reveal the possibilities and limits of the social engineering enterprise, and suggest that historians of technology might consider how the history of psychology and other psy-disciplines can deepen conceptualizations of the relationships among the psychological, the social, and the technological in this period. (PsycINFO Database Record


Novel Interface for High-Throughput Analysis of Biotherapeutics by Electrospray Mass Spectrometry.
With the rapid rise of therapeutic antibodies and antibody-drug conjugates, significant investments have been made in developing workflows that utilize mass spectrometry to detect these intact molecules, the large fragments generated by their selective digestion, and the peptides generated by traditional proteomics workflows. The resultant data is used to gain insight into a wide range of parameters, including primary sequence, disulfide bonding, glycosylation patterns, biotransformation, and more. However, many of the technologies utilized to couple these workflows to mass spectrometers have significant limitations that force nonoptimal modifications to upstream sample preparation steps, limit the throughput of high-volume workflows, and prevent the harmonization of diverse experiments onto a single hardware platform. Here, we describe a new analytical platform that enables direct and high-throughput coupling to electrospray ionization mass spectrometry. The SampleStream platform is compatible with both native and denaturing electrospray, operates with a throughput of up to 15 s/sample, provides extensive concentration of dilute samples, and affords similar sensitivity to comparable liquid chromatographic methods.


Medullary raphe: a new site for vagally mediated stimulation of gastric motility in cats.
Thyrotropin-releasing hormone (TRH) is clearly implicated in the control of gastric function via interactions in the dorsal motor nucleus of the vagus (DMV) of the cat. The source of the TRH innervation of the DMV is important to determine because this region could be of importance in control of gastric function. TRH-immunoreactive (ir) neurons are located in the raphe obscurus (Ro), raphe pallidus (Rp), and raphe magnus (Rm). Retrograde tracer applied to the DMV resulted in the most numerous labeled neurons in the caudal Ro and Rp in the same region where TRH-ir neurons are located. To address the question whether DMV-projecting neurons in the raphe subnuclei play a role in control of gastric motility, the following experiments were performed in alpha-chloralose-anesthetized cats while recording pyloric motility and blood pressure. Microinjection of a cell body excitant L-glutamate (44-200 nl, 0.5 M) into the caudal Ro and Rp in 15 experiments produced significant increases in pyloric minute motility index (MMI) of 4.9 +/- 1.5 (from 1.6 +/- 0.7 preinjection to 6.5 +/- 1.8 postinjection, P less than 0.05). Mean blood pressure (MBP) decreased significantly in these animals by 12 +/- 7 mmHg (from 100 +/- 6 to 88 +/- 8 mmHg, P less than 0.05). Saline microinjection in the same sites in seven cases resulted in no significant change in pyloric MMI (-1.0 +/- 0.8) or MBP (-4 +/- 11 mmHg). In five of these experiments, a second microinjection of L-glutamate (132-240 nl) was performed into the caudal Ro and Rp after spinal cord transection. This resulted in a significant increase in pyloric MMI of 3.3 +/- 0.9 (from 1.0 +/- 0.5 preinjection to 4.3 +/- 1.1 postinjection, P less than 0.05) but no change in MBP (+1 +/- 1 mmHg). Bilateral vagotomy resulted in the abrupt cessation of the pyloric response to caudal Ro and Rp stimulation. Microinjection of L-glutamate into the rostral Rp and caudal Rm in nine experiments resulted in no significant changes in pyloric MMI (-0.4 +/- 0.8) or MBP (-10 +/- 11 mmHg). These data indicate that a population of neurons in the caudal raphe nuclei, which may contain TRH, project to the DMV. In addition, excitation of these neurons causes an increase in gastric motility that is not caused by inhibition of sympathetic outflow to the gut but rather by excitation of vagal neurons in the DMV.


Virion Display: A High-Throughput Method to Express Functional Membrane Proteins.
Transmembrane proteins are responsible for many critical cellular functions and represent one of the largest families of drug targets. However, these proteins, especially multipass transmembrane proteins, are difficult to study because they must be embedded in a lipid bilayer to maintain their native conformations. The development of the virion display (VirD) technology enables transmembrane proteins to be integrated into the viral envelope of herpes simplex virus 1 (HSV-1). Combining high-throughput cloning, expression, and purification techniques, VirD technology has been applied to the largest set of human transmembrane proteins, namely G-protein-coupled receptors, and has allowed the identification of interactions that are both specific and functional. This article describes the procedures to integrate an open reading frame for any transmembrane protein into the HSV-1 genome and produce recombinant HSV-1 virus to ultimately generate pure VirD virions for biological and pharmaceutical studies. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Gateway cloning of transmembrane proteins Support Protocol 1: Ethanol precipitation of bacterial artificial chromosomal DNA Support Protocol 2: Preparation of competent cells Basic Protocol 2: Production of recombinant HSV-1 virions.


METTL15 introduces N4-methylcytidine into human mitochondrial 12S rRNA and is required for mitoribosome biogenesis.
Post-transcriptional RNA modifications, the epitranscriptome, play important roles in modulating the functions of RNA species. Modifications of rRNA are key for ribosome production and function. Identification and characterization of enzymes involved in epitranscriptome shaping is instrumental for the elucidation of the functional roles of specific RNA modifications. Ten modified sites have been thus far identified in the mammalian mitochondrial rRNA. Enzymes responsible for two of these modifications have not been characterized. Here, we identify METTL15, show that it is the main N4-methylcytidine (m4C) methyltransferase in human cells and demonstrate that it is responsible for the methylation of position C839 in mitochondrial 12S rRNA. We show that the lack of METTL15 results in a reduction of the mitochondrial de novo protein synthesis and decreased steady-state levels of protein components of the oxidative phosphorylation system. Without functional METTL15, the assembly of the mitochondrial ribosome is decreased, with the late assembly components being unable to be incorporated efficiently into the small subunit. We speculate that m4C839 is involved in the stabilization of 12S rRNA folding, therefore facilitating the assembly of the mitochondrial small ribosomal subunits. Taken together our data show that METTL15 is a novel protein necessary for efficient translation in human mitochondria.


Seasonal variations of cough reflex sensitivity in elite athletes training in cold air environment.
Exercise-induced cough is common among athletes. Athletes training in cold air often report an increasingly troublesome cough during the winter season. Chronic airway irritation or inflammation may increase the sensory response of cough receptors. The aim of this study was to evaluate the seasonal variability of cough reflex sensitivity to capsaicin in elite athletes. Fifty-three elite winter athletes and 33 sedentary subjects completed a respiratory questionnaire and a capsaicin provocation test during the summer, fall, and winter. Allergy skin prick tests, spirometry, eucapnic voluntary hyperpnea test (EVH), methacholine inhalation test (MIT), and induced sputum analysis were also performed. In athletes, the prevalence of cough immediately after exercise was high, particularly during winter. Athletes often showed a late occurrence of cough between 2-8 h after exercise. The cough reflex sensitivity to capsaicin was unchanged through the seasons in both athletes and non-athlete subjects. No significant correlations were found in groups between cough reflex sensitivity to capsaicin and the number of years in sport training, the number of hours of training per week, EVH response (% fall in FEV1), airway responsiveness to methacholine (PC20), airway inflammation or atopy. The prevalence of cough immediately and a few hours after exercise is high in athletes and more frequently reported during winter. However, cough does not seem to be associated with cough reflex hypersensitivity to capsaicin, bronchoconstriction, or airway inflammation in the majority of athletes.


Obesity and cancer: pathophysiological and biological mechanisms.
Excess body weight (overweight and obesity) is characterized by chronic hyperinsulinaemia and insulin resistance, and is implicated both in cancer risk and cancer mortality. The list of cancers at increased risk of development in an "obesogenic" environment include common adult cancers such as endometrium, post-menopausal breast, colon and kidney, but also less common malignancies such as leukaemia, multiple myeloma, and non-Hodgkin's lymphoma. The pathophysiological and biological mechanisms underpinning these associations are only starting to be understood. Insulin resistance is at the heart of many, but there are several other candidate systems including insulin-like growth factors, sex steroids, adipokines, obesity-related inflammatory markers, the nuclear factor kappa beta (NF-kappa B) system and oxidative stresses. With such as diversity of obesity-related cancers, it is unlikely that there is a "one system fits all" mechanism. While public health strategies to curb the spread of the obesity epidemic appear ineffective, there is a need to better understand the processes linking obesity and cancer as a pre-requisite to the development of new approaches to the prevention and treatment of obesity-related cancers.


[Features of distribution of free and esterified cholesterol in the epidermis, biological membranes and plasma lipoproteins in psoriasis].
The parameters of cholesterol (CS) metabolism in the epidermis, blood plasma lipoproteins, and red cell membranes were assessed in patients with psoriasis. The levels of free and total CS in extracts from the surface of epidermis, in plasma lipoproteins, and in plasma triglycerides were measured, and the molar ratio CS/ phospholipids (CS/PL index) in red cell membranes assessed in 194 patients with psoriasis of different severity. The levels of free and total CS in the epidermis were found to increase as the psoriatic process grew in severity. These shifts manifested even in apparently intact skin sites, which indicated the primary nature of total-system disorders. As for the plasma lipids, statistically reliable changes were observed only for the relative level of free cholesterol of the plasma and, most of all, high-density lipoproteins. These values for the mean parameters in the five groups of examinees correlated (r from 0.71 to 0.92) with the levels of total and free CS in the epidermis. Contrary to it, the levels of total CS and triglycerides in the plasma had a tendency to decrease as the severity of the psoriatic process augmented. Such a trend was statistically reliable for the CS/PL index, which increased in all the patients except the most grave, this increase being the more, the less grave the disease was; this was manifested in an inverse correlation (r = 0.9, on average) with the epidermal CS. The detected uncommon relationship between CS values in the epidermis, plasma lipoproteins, and biological membranes indicates heretofore unknown features of metabolic disorders in psoriasis.


Disaster education and preparedness in the acute care setting: A cross sectional survey of operating theatre nurse's disaster knowledge and education.
Operating theatre services can be heavily relied upon during mass casualty disaster events, which require nurses to have adequate training and education of hospital disaster management plans to respond appropriately. The evidence-base of disaster preparedness in the acute setting is limited, particularly with regard to operating theatre nurses. Explore operating theatre nurse's disaster knowledge of their role in a mass casualty event, and identify the preferred mode of disaster education and training to improve disaster preparedness. A cross-sectional research design was employed with data collected using a survey tool. The research was undertaken on operating theatre nurses in a tertiary hospital in Victoria, Australia. The participants in this research included 53 operating theatre nurses, 51 Registered Nurses and 2 Enrolled Nurses. The survey was based on a disaster questionnaire for emergency department nurses from South Australia, exploring knowledge and preparedness for disaster response in the acute setting, and altered to be specific and relevant to the operating theatre environment and broadened to focus on the training needs of perioperative nurses. The survey of 53 operating theatre nurses identified that few had previous disaster experience (19.9%). The majority of respondents were aware of their disaster management policy (Code Brown policy) (94.1%), of reporting lines, and appropriate triage (80.4%). However, a significant number of nurses (50.9%) stated that in the event of a disaster they would "call work to see if [they needed] additional assistance" compared with 43.4% of respondents "wait to be contacted by a manager/floor coordinator" as was policy. Finally, disaster nursing general knowledge amongst staff was poor; a mean of 1.79 (SD = 1.20) correct answers out of a possible 7. This study highlights that disaster education and training methods for disasters be specific to the role required by nurses and all staff during a disaster activation; training drills are preferred although face-to-face education is practical.


Risk of hair loss with different antidepressants: a comparative retrospective cohort study.
The aim of this study was to quantify the risk of hair loss with different antidepressants. A retrospective cohort study design using a large health claims database in the USA from 2006 to 2014 was utilized. A cohort of new user and mutually exclusive users of fluoxetine, fluvoxamine, sertraline, citalopram, escitalopram, paroxetine, duloxetine, venlafaxine, desvenlafaxine, and bupropion were followed to the first diagnosis of alopecia. The cohort was comprised of 1 025 140 new users of fluoxetine, fluvoxamine, sertraline, citalopram, escitalopram, paroxetine, duloxetine, venlafaxine, desvenlafaxine, and bupropion, with sertraline the most commonly prescribed (N=190 227) and fluvoxamine (N=3010) the least prescribed. Compared with bupropion, all other antidepressants had a lower risk of hair loss, with fluoxetine and paroxetine having the lowest risk [hazard ratio (HR)=0.68, 95% confidence interval (CI): 0.63-0.74, HR=0.68, 95% CI: 0.62-0.74, respectively] and fluvoxamine having the highest risk (HR=0.93, 95% CI: 0.64-1.37). Compared with fluoxetine, bupropion had the highest risk of hair loss (HR=1.46, 95% CI: 1.35-1.58, number needed to harm=242 for 2 years) and paroxetine had the lowest risk (HR=0.99, 95% CI: 0.90-1.09). The results of this large population-based cohort study suggest an increase in the risk of hair loss with bupropion compared with selective serotonin reuptake inhibitors and selective norepinephrine reuptake inhibitors, whereas paroxetine had the lowest risk.


Solid-state hydrogen rich boron-nitrogen compounds for energy storage.
Boron compounds have a rich history in energy storage applications, ranging from high energy fuels for advanced aircraft to hydrogen storage materials for fuel cell applications. In this review we cover some of the aspects of energy storage materials comprised of electron-poor boron materials combined with electron-rich nitrogen elements with the goal of moderate temperature release of hydrogen. The parent compounds of ammonium borohydride, ammonia borane, and diammoniate of diborane provide approaches for storing high gravimetric and volumetric densities of hydrogen. Here we provide a review with a historical perspective and current developments in the area of solid state B and N containing compounds. This review highlights developments in synthesis of ammonia borane and its derivatives over the last 80 years. Thermodynamics and kinetics of hydrogen release in the solid state are discussed. By changing either substituents on the boron and nitrogen atoms or the physical environment by embedding in mesoporous scaffolds, the thermodynamics can be modified to reduce the exothermicity of hydrogen release and minimize formation of volatile impurities. Several mechanistic studies are reviewed identifying the key distinctions between homopolar and heteropolar H2 release. Strategies for economical and efficient regeneration of the hydrogen storage materials via chemical transformation are critically reviewed. The limited efficiency of these chemical regeneration has limited some of the potential applications.


Measles in Chimanimani Zimbabwe.
A mini outbreak of measles in Chimanimani District in Zimbabwe where the measles immunisation coverage is about 79% is reported. Although many of the cases occurred in susceptible non-immunised children of the vaPostori (Apostolic) religious sects, a large number was in teenagers and adults. The importance of health education in increasing community participation in immunisation is emphasised. There is need to chart out a strategy to deal with a projected pool of persons susceptible to measles as our coverage rates continue to increase.


Circulatory TGF-beta1 is significantly higher in early stage of pulmonary sarcoidosis.
Introduction: The pathogenesis of pulmonary fibrosis in sarcoidosis is not known. We hypothesized that higher levels of circulatory growth factors are present in early stages of pulmonary sarcoidosis and may be associated with pulmonary fibrosis. Methods: Age and sex-matched subjects with sarcoidosis stage 0-1 (n=18), stage 4-5 (n=13) and healthy controls (n=5) had their serum TGF-β1, FGF, and VEGF levels measured as well as their gene expressions determined in peripheral blood mononuclear cells. Results: TGF-β1 levels were significantly higher in patients with stage 0-1 sarcoidosis compared with normal healthy control patients (25,488 vs. 13800 pg/ml, P=0.05). Patients with sarcoidosis stage 4 had a 1.3-fold higher peripheral blood mononuclear cells (PBMC) gene expression of TGF-β1 compared with subjects at stage 0-1 (P= 0.041). The serum levels of FGF, and VEGF had a trend towards higher levels in sarcoidosis subjects compared to normal controls. Conclusion: These results suggest that cell growth factors levels are high in early stages of sarcoidosis. These findings should be validated in larger studies. (Sarcoidosis Vasc Diffuse Lung Dis 2018; 35: 213-217).


Towards a generic procedure for the detection of relevant contaminants from waste electric and electronic equipment (WEEE) in plastic food-contact materials: a review and selection of key parameters.
Recently, traces of brominated flame retardants (BFRs) have been detected in black plastic food-contact materials (FCMs), indicating the presence of recycled plastics, mainly coming from waste electric and electronic equipment (WEEE) as BFRs are one of the main additives in electric applications. In order to evaluate efficiently and preliminary in situ the presence of WEEE in plastic FCMs, a generic procedure for the evaluation of WEEE presence in plastic FCMs by using defined parameters having each an associated importance level has been proposed. This can be achieved by combining parameters like overall bromine (Br) and antimony (Sb) content; additive and reactive BFR, rare earth element (REE) and WEEE-relevant elemental content and additionally polymer purity. In most of the cases, the WEEE contamination could be confirmed by combining X-ray fluorescence (XRF) spectrometry and thermal desorption/pyrolysis gas chromatography-mass spectrometry (GC-MS) at first. The Sb and REE content did not give a full confirmation as to the source of contamination, however for Sb the opposite counts: Sb was joined with elevated Br signals. Therefore, Br at first followed by Sb were used as WEEE precursors as both elements are used as synergetic flame-retardant systems. WEEE-specific REEs could be used for small WEEE (sWEEE) confirmation; however, this parameter should be interpreted with care. The polymer purity by Fourier-transform infrared spectrometer (FTIR) and pyrolysis GC-MS in many cases could not confirm WEEE-specific contamination; however, it can be used for purity measurements and for the suspicion of the usage of recycled fractions (WEEE and non-WEEE) as a third-line confirmation. To the best of our knowledge, the addition of WEEE waste to plastic FCMs is illegal; however, due to lack on screening mechanisms, there is still the breakthrough of such articles onto the market, and, therefore, our generic procedure enables the quick and effective screening of suspicious samples.


Hepatic expression of the SPOT 14 (S14) paralog S14-related (Mid1 interacting protein) is regulated by dietary carbohydrate.
The Spot 14 (S14) gene is rapidly up-regulated by signals that induce lipogenesis such as enhanced glucose metabolism and thyroid hormone administration. Previous studies in S14 null mice show that S14 is required for normal lipogenesis in the lactating mammary gland, but not the liver. We speculated that the lack of a hepatic phenotype was due to the expression of a compensatory gene. We recently reported that this gene is likely an S14 paralog that we named S14-Related (S14-R). S14-R is expressed in the liver, but not in the mammary gland. If S14-R compensates for the absence of S14 in the liver, we hypothesized that, like S14, S14-R expression should be glucose responsive. Here, we report that hepatic S14-R mRNA levels increase with high-carbohydrate feeding in mice or within 2 h of treating cultured hepatocytes with elevated glucose. A potential carbohydrate response element (ChoRE) was identified at position -458 of the S14-R promoter. Deletion of or point mutations within the putative S14-R ChoRE led to 50-95% inhibition of the glucose response. Gel-shift analysis revealed that the glucose-activated transcription complex carbohydrate responsive element-binding protein/Max-like protein X (Mlx) binds to the S14-R ChoRE. Finally, S14-R glucose induction is completely blocked when a dominant-negative form of Mlx is overexpressed in primary hepatocytes. In conclusion, our results indicate that the S14-R gene is a glucose-responsive target of carbohydrate responsive element-binding protein/Mlx and suggest that the S14-R protein is a compensatory factor, at least partially responsible for the normal liver lipogenesis observed in the S14 null mouse.


Survey of Australian practitioners' provision of healthy lifestyle advice to clients who are obese.
Obesity is a global issue, with healthcare practitioners increasingly involved in clinical interactions with people who are overweight or obese. These interactions are opportunities to provide evidence-based healthy lifestyle advice, and impact on public health. This study used a cross-sectional survey of Australian healthcare practitioners to investigate what influenced the provision of healthy lifestyle advice to obese and overweight clients. A modified theory of planned behavior was used to explore knowledge translation processes. Knowledge translation was linked to three factors: (i) a healthcare practitioner's education and confidence in the currency of their knowledge; (ii) personal characteristics - whether they accepted that providing this advice was within their domain of practice; and (iii) the existence of organizational support structures, such as access to education, and best practice guidelines. To fulfill the potential role healthcare practitioners can play in the provision of evidence-based health promotion advice requires organizations to provide access to practice guidelines and to instill a belief in their workforce that this is a shared professional domain.


Reversal of chloroquine resistance in Plasmodium falciparum by verapamil.
The parasite Plasmodium falciparum, like neoplastic cells, develops resistance to multiple structurally unrelated drugs. If the mechanisms by which P. falciparum and neoplastic cells become resistant are similar, then it may be possible to reverse the resistance in the two types of cells by the same pharmacological agents. Verapamil, a calcium channel blocker, completely reversed chloroquine resistance in two chloroquine-resistant P. falciparum clones from Southeast Asia and Brazil. Verapamil reversed chloroquine resistance at the same concentration (1 X 10(-6)M) as that at which it reversed resistance in multidrug-resistant cultured neoplastic cells. This same concentration of verapamil had no effect on chloroquine-sensitive parasites. Hence, chloroquine resistance in P. falciparum may fit the criteria for the multidrug-resistant phenotype.


Beneficial effect of the soluble guanylyl cyclase stimulator BAY 41-2272 on impaired penile erection in db/db-/- type II diabetic and obese mice.
Type 2 diabetes mellitus (DM2) and obesity are major risk factors for erectile dysfunction (ED). In diabetes, increased oxidative stress leads to decreased nitric oxide (NO) bioavailability, and diabetic patients appear to be less responsive to conventional therapy with phosphodiesterase type 5 inhibitors. We investigated whether the soluble guanylyl cyclase stimulator BAY 41-2272 (5-cyclopropyl-2-[1-(2-fluoro-benzyl)-1H-pyrazolo[3,4-b]pyridine-3-yl]pyrimidin-4ylamine) is effective in improving impaired corpus cavernosum (CC) relaxation in obese DM2 mice by reducing oxidative stress. Adult db/db(-/-) mice or their lean db(/+) littermates were used to assess vascular function, cGMP levels, antioxidant status, NADPH oxidase expression, and superoxide formation in the absence or presence of BAY 41-2272. Results showed that BAY 41-2272 (10(-8) to 10(-5) M) potently relaxed CC from db(/+) or db/db(-/-) mice in a similar manner. BAY 41-2272 significantly enhanced both endothelium-dependent and nitrergic relaxation induced by electrical field stimulation (EFS), and improved the impaired relaxation to acetylcholine and EFS in the diabetic animals in a concentration-dependent manner (10(-8) to 10(-7) M). BAY 41-2272 increased cGMP levels and potentiated relaxation responses to exogenous NO in CC. Total antioxidant status was reduced in plasma and urine whereas expression of vascular NADPH oxidase subunits (gp91phox, p22phox, and p47phox) was increased in the CC of db/db(-/-) mice, suggesting a state of oxidative stress. These effects were prevented by BAY 41-2272 in a concentration-dependent manner. These results suggest that BAY 41-2272 improves CC relaxation in db/db(-/-) mice by increasing cGMP and augmenting antioxidant status, making this drug is a potential novel candidate to treat ED.


Electrochemical behavior of quinapril and its determination in pharmaceutical formulations by square-wave voltammetry at a mercury electrode.
The electrochemical behavior of the antihypertensive drug quinapril was investigated at a hanging mercury drop electrode using different voltammetric techniques such as cyclic voltammetry, square-wave voltammetry and chronoamperometry. A simple and sensitive square-wave voltammetric method for the electrochemical analysis of quinapril in its pharmaceutical formulations was developed and validated. The experimental and instrumental parameters affecting the peak current of quinapril were investigated. Various buffers such as Britton Robinson, borate and phosphate buffers at different pH values (3.0-11.0) were examined as supporting electrolyte. The optimum conditions were obtained using Britton Robinson buffer at pH 10.0 and frequency: 50 Hz, scan increment: 4 mV and pulse amplitude: 25 mV. A well-defined peak current was observed at the hanging mercury drop electrode at -1100 mV vs. Ag/AgCl reference electrode. This proposed method was validated by evaluating linearity, sensitivity, repeatability, accuracy, precision, selectivity, recovery, robustness and ruggedness. The linear calibration range was 0.50-8.68 microg mL-' (r = 0.9992). The detection and quantification limits of this method were 0.22 and 0.50 Ctg mL(-1) and intra-day and inter-day precision were between 0.81-4.32% (n = 7), respectively. The developed method was applied successfully for the determination of quinapril in its tablet dosage forms. The average amount of quinapril in tablets was found as 20.26 +/- 0.12 with RSD of 1.60% for 20 mg tabletsand 40.55 +/- 0.23 with RSD of 1.52% for 40 mg tablets.


Use of performance metrics for the measurement of universal coverage for maternal care in Mexico.
This study provides evidence for those working in the maternal health metrics and health system performance fields, as well as those interested in achieving universal and effective health care coverage. Based on the perspective of continuity of health care and applying quasi-experimental methods to analyse the cross-sectional 2009 National Demographic Dynamics Survey (n = 14 414 women), we estimated the middle-term effects of Mexico's new public health insurance scheme, Seguro Popular de Salud (SPS) (vs women without health insurance) on seven indicators related to maternal health care (according to official guidelines): (a) access to skilled antenatal care (ANC); (b) timely ANC; (c) frequent ANC; (d) adequate content of ANC; (e) institutional delivery; (f) postnatal consultation and (g) access to standardized comprehensive antenatal and postnatal care (or the intersection of the seven process indicators). Our results show that 94% of all pregnancies were attended by trained health personnel. However, comprehensive access to ANC declines steeply in both groups as we move along the maternal healthcare continuum. The percentage of institutional deliveries providing timely, frequent and adequate content of ANC reached 70% among SPS women (vs 64.7% in the uninsured), and only 57.4% of SPS-affiliated women received standardized comprehensive care (vs 53.7% in the uninsured group). In Mexico, access to comprehensive antenatal and postnatal care as defined by Mexican guidelines (in accordance to WHO recommendations) is far from optimal. Even though a positive influence of SPS on maternal care was documented, important challenges still remain. Our results identified key bottlenecks of the maternal healthcare continuum that should be addressed by policy makers through a combination of supply side interventions and interventions directed to social determinants of access to health care.


Structure-based design of nonnatural ligands for the HLA-B27 protein.
X-ray studies as well as structure-activity relationships indicate that the central part of class I MHC-binding nonapeptides represents the main interaction site for a T cell receptor. In order to rationally manipulate T cell epitopes, several nonpeptidic spacer have been designed from the X-ray structure of a MHC-peptide complex and substituted for the T cell receptor-binding part of several antigenic peptides. The binding of the modified epitopes to the HLA-B*2705 protein was studied by an in vitro stabilisation assay and the thermal stability of all complexes examined by circular dichroism spectroscopy. Depending on their chemical nature and length, the introduced spacers may be classified into two categories. Monofunctional spacers (11-amino undecanoate, (R)-3-hydroxybutyrate trimer) simply link two anchoring peptide positions (P3 and P9) but loosely contact the MHC binding groove, and thus decrease more or less the affinity of the altered epitopes to HLA-B*2705. Bifunctional spacers ((R)-3-hydroxybutyrate and beta-homoalanine combinations) not only bridges the two distant anchoring amino acids but also strongly interact with the binding cleft and lead to an increase in binding to the MHC protein. The presented modified ligands constitute interesting tools for perturbing the T cell response to the parent antigenic peptide.


Immune complexes in cystic fibrosis.
Circulating immune complexes were detected in serum and sputum of patients with cystic fibrosis (C.F.). There were extensive deposits of immunoglobulins and complement immune complexes in several of the C.F. organs, especially the respiratory and gastrointestinal tracts, but not in the kidneys. Significant concentrations of IgG and of complement complexes could be eluted from the lungs of the C.F. patients but not from those of controls. Studies involving immunoabsorption, autoradiography, and molecular sieving through Sephadex G-200 columns identified both bovine serum albumin and staphylococcal alpha-haemolysin as two of the antigens present in the immune complexes. The sedimentation constant of the immune complexes was about 8S to 11S. The clinical significance of these immune complexes and the wide variety of antibodies detected in C.F. patients are discussed.


Survival and growth of Escherichia coli O157:H7 on salad vegetables.
The influence of modified-atmosphere packaging, storage temperature, and time on survival and growth of Escherichia coli O157:H7 inoculated onto shredded lettuce, sliced cucumber, and shredded carrot was determined. Growth of psychotrophic and mesophilic microorganisms and changes in pH and sensory qualities of vegetables, as judged by subjective evaluation, were also monitored. Packaging under an atmosphere containing 3% oxygen and 97% nitrogen had no apparent effect on populations of E. coli O157:H7, psychotrophs, or mesophiles. Populations of viable E. coli O157:H7 declined on vegetables stored at 5 degrees C and increased on vegetables stored at 12 and 21 degrees C for up to 14 days. The most rapid increases in populations of E. coli O157:H7 occurred on lettuce and cucumbers stored at 21 degrees C. These results suggest that an unknown factor(s) associated with carrots may inhibit the growth of E. coli O157:H7. The reduction in pH of vegetables was correlated with initial increases in populations of E. coli O157:H7 and naturally occurring microfloras. Eventual decreases in E. coli O157:H7 in some samples, e.g., those stored at 21 degrees C, are attributed to the toxic effect of accumulated acids. Changes in visual appearance of vegetables were not influenced substantially by growth of E. coli O157:H7. The ability of E. coli O157:H7 to growth on raw salad vegetables subjected to processing and storage conditions simulating those routinely used in commercial practice has been demonstrated.


The fragile spirituality of parents whose children died in the pediatric intensive care unit.
Spiritual care is recognized as a relevant dimension of health care. In the context of pediatric palliative end-of-life care, spirituality entails more than adhering to a spiritual worldview or religion. Interviews with parents whose critically ill child died in the pediatric intensive care unit revealed features of a spirituality that is fragmentary and full of contradictions. This type of spirituality, which we refer to as fragile, speaks of parents' connectedness with the deceased child and the hope of some kind of reuniting after one's own death. Acknowledging that fragments of spirituality can be part of parents' experiences in their child's end-of-life stage can be a meaningful contribution to compassionate care.


Isoflavone supplementation in postmenopausal women does not affect leukocyte LDL receptor and scavenger receptor CD36 expression: A double-blind, randomized, placebo-controlled trial.
Isoflavones are discussed to improve serum lipoproteins and body composition and to reduce cardiovascular disease risk in postmenopausal women (PMW). LDL receptors (LDLR) and scavenger receptor CD36 (CD36) play a pivotal role in the regulation of plasma LDL-cholesterol concentrations (LDL-chol). We investigated the impact of isoflavones on the receptor expression of both receptors in leukocytes of PMW. A randomized, double-blind, placebo-controlled trial in parallel design was conducted to assess the effects of an isoflavone-enriched soy extract (117.4 mg/day isoflavone aglycone equivalents) for 12 weeks on serum LDL-chol, LDLR, and CD36 expression on leukocytes in 170 healthy PMW. Baseline and after 12 weeks, blood lipid concentrations, anthropometric data and body composition were determined. Receptor expression on leukocytes was measured by means of flow cytometry. After the intervention, no significant differences were found for LDLR and CD36 expression on leukocytes. A significant increase of serum LDL-chol was shown for the isoflavone group (p = 0.03) after 12 weeks. Body fat content and VAT were not affected. Isoflavone supplementation for 12 weeks did not change LDLR and CD36 expression on leukocytes of PMW and did not affect body fat content and visceral adipose tissue (VAT), but slightly increased serum LDL-chol.


An essential role of Sam50 in the protein sorting and assembly machinery of the mitochondrial outer membrane.
The preprotein translocase of the outer mitochondrial membrane (TOM complex) contains one essential subunit, the channel Tom40. The assembly pathway of the precursor of Tom40 involves the TOM complex and the sorting and assembly machinery (SAM complex) with the non-essential subunit Mas37. We have identified Sam50, the second essential protein of the mitochondrial outer membrane. Sam50 contains a beta-barrel domain conserved from bacteria to man and is a subunit of the SAM complex. Yeast mutants of Sam50 are defective in the assembly pathways of Tom40 and the abundant outer membrane protein porin, while the import of matrix proteins is not affected. Thus the protein sorting and assembly machinery of the mitochondrial outer membrane involves an essential, conserved protein.


Sequence analysis of the major outer membrane protein gene from Chlamydia trachomatis serovar L2.
The structural gene for the major outer membrane protein (MOMP) from Chlamydia trachomatis was cloned and sequenced. A lambda gt11 recombinant (lambda gt11/L2/33) that contains a portion of the MOMP coding sequence was used to probe a lambda 1059 library constructed from DNA obtained from C. trachomatis serovar L2. Selected lambda 1059 recombinants were mapped with endonuclease restriction enzymes. The MOMP gene was mapped to the 5' end of a BamHI fragment of approximately 9 kilobases. Contiguous endonuclease restriction fragments identified within this region permitted the selection of specific fragments for subcloning and DNA sequencing. The MOMP gene consisted of a 1,182-base-pair open reading frame that encoded 394 amino acids and ended with three stop codons. The known amino-terminal amino acid was preceded by 22 amino acids whose sequence was compatible with a leader or signal sequence. The primary structure of MOMP determined from the translated DNA sequence demonstrated nine cysteine residues and a remarkably homogeneous distribution of charged and hydrophobic residues.


Molecular testing in the diagnosis and management of chronic hepatitis B.
Hepatitis B virus (HBV) is an enveloped virus with a small (3.2-kb) partially double-stranded DNA genome that causes acute and chronic infections. The impact of these infections on public health worldwide is enormous, with an estimated prevalence of 2 billion acute infections and 360 million chronic infections globally. This review focuses on chronic hepatitis B and the molecular assays used in its diagnosis and management. Background information, including that about features of the hepatitis B virion, viral replication, and epidemiology of infection, that is important for understanding chronic hepatitis B and molecular diagnostic tests for HBV is provided. To facilitate an understanding of the utility of molecular testing for chronic hepatitis B, the four stages of chronic hepatitis B infection that are currently recognized, as well as an additional entity, occult hepatitis B, that can be diagnosed only by sensitive nucleic acid amplification methods, are reviewed in detail, including available therapeutic agents. The molecular diagnostic content focuses on tests for HBV DNA quantification, genotyping, and mutation detection (including precore/core promoter and antiviral resistance mutations). The discussion of these tests encompasses their current utility and performance characteristics, drawing from current clinical guidelines and other studies from the literature. In recognition of the continual evolution of this field, the final section describes emerging molecular markers with future diagnostic potential.


Recruitment of RED-SMU1 complex by Influenza A Virus RNA polymerase to control Viral mRNA splicing.
Influenza A viruses are major pathogens in humans and in animals, whose genome consists of eight single-stranded RNA segments of negative polarity. Viral mRNAs are synthesized by the viral RNA-dependent RNA polymerase in the nucleus of infected cells, in close association with the cellular transcriptional machinery. Two proteins essential for viral multiplication, the exportin NS2/NEP and the ion channel protein M2, are produced by splicing of the NS1 and M1 mRNAs, respectively. Here we identify two human spliceosomal factors, RED and SMU1, that control the expression of NS2/NEP and are required for efficient viral multiplication. We provide several lines of evidence that in infected cells, the hetero-trimeric viral polymerase recruits a complex formed by RED and SMU1 through interaction with its PB2 and PB1 subunits. We demonstrate that the splicing of the NS1 viral mRNA is specifically affected in cells depleted of RED or SMU1, leading to a decreased production of the spliced mRNA species NS2, and to a reduced NS2/NS1 protein ratio. In agreement with the exportin function of NS2, these defects impair the transport of newly synthesized viral ribonucleoproteins from the nucleus to the cytoplasm, and strongly reduce the production of infectious influenza virions. Overall, our results unravel a new mechanism of viral subversion of the cellular splicing machinery, by establishing that the human splicing factors RED and SMU1 act jointly as key regulators of influenza virus gene expression. In addition, our data point to a central role of the viral RNA polymerase in coupling transcription and alternative splicing of the viral mRNAs.


Nutritional assessment by ultrasound of the rectus femoris and preperitoneal adipose tissue as predictors of hospitalized patient complications.
Malnutrition and sarcopenia are associated with increased risk of adverse events and worse outcome in hospitalised patients. Ultrasonography is a useful tool in the assessment of body composition. Twenty-three patients admitted to a hospital ward were recruited. Ultrasonographic variables of muscle tissue and adipose tissue were collected, as well as their variation during admission. These were correlated with anthropometric, clinical and analytical data. We observed a correlation of ultrasound measurements with length of hospitalisation (maximum preperitoneal fat -0.585, rectus femoris thickness - RF -0.539, RF area -0.540), frailty (RF -0.599) and dependency (RF 0.628). RF contractile capacity correlated with reported weekly exercise (0.642). RF and preperitoneal fat thickness correlate with the number of days of admission and functional capacity, emerging as prognostic variables.


Thermodynamics of transport through the ammonium transporter Amt-1 investigated with free energy calculations.
Amt-1 from Archaeoglobus fulgidus (AfAmt-1) belongs to the Amt/Rh family of ammonium/ammonia transporting membrane proteins. The transport mode and the precise microscopic permeation mechanism utilized by these proteins are intensely debated. Open questions concern the identity of the transported substrate (ammonia and/or ammonium) and whether the transport is passive or active. To address these questions, we studied the overall thermodynamics of the different transport modes as a function of the environmental conditions. Then, we investigated the thermodynamics of the underlying microscopic transport mechanisms with free energy calculations within a continuum electrostatics model. The formalism developed for this purpose is of general utility in the calculation of binding free energies for ligands with multiple protonation forms or other binding forms. The results of our calculations are compared to the available experimental and theoretical data on Amt/Rh proteins and discussed in light of the current knowledge on the physiological conditions experienced by microorganisms and plants. We found that microscopic models of electroneutral and electrogenic transport modes are in principle thermodynamically viable. However, only the electrogenic variants have a net thermodynamic driving force under the physiological conditions experienced by microorganisms and plants. Thus, the transport mechanism of AfAmt-1 is most likely electrogenic.


Deep Evolutionary History of the Phox and Bem1 (PB1) Domain Across Eukaryotes.
Protein oligomerization is a fundamental process to build complex functional modules. Domains that facilitate the oligomerization process are diverse and widespread in nature across all kingdoms of life. One such domain is the Phox and Bem1 (PB1) domain, which is functionally well-studied in the animal kingdom. However, beyond animals, neither the origin nor the evolutionary patterns of PB1-containing proteins are understood. While PB1 domain proteins have been found in other kingdoms including plants, it is unclear how these relate to animal PB1 proteins. To address this question, we utilized large transcriptome datasets along with the proteomes of a broad range of species. We discovered eight PB1 domain-containing protein families in plants, along with four each in Protozoa and Fungi and three families in Chromista. Studying the deep evolutionary history of PB1 domains throughout eukaryotes revealed the presence of at least two, but likely three, ancestral PB1 copies in the Last Eukaryotic Common Ancestor (LECA). These three ancestral copies gave rise to multiple orthologues later in evolution. Analyzing the sequence and secondary structure properties of plant PB1 domains from all the eight families showed their common ubiquitin β-grasp fold, despite poor sequence identity. Tertiary structural models of these plant PB1 families, combined with Random Forest based classification, indicated family-specific differences attributed to the length of PB1 domain and the proportion of β-sheets. Thus, this study not only identifies novel PB1 families, but also provides an evolutionary basis to understand their diverse functional interactions.


Assessment of thermal effects of interstitial laser phototherapy on mammary tumors using proton resonance frequency method.
Laser immunotherapy (LIT) uses a synergistic approach to treat cancer systemically through local laser irradiation and immunological stimulation. Currently, LIT utilizes dye-assisted noninvasive laser irradiation to achieve selective photothermal interaction. However, LIT faces difficulties treating deeper tumors or tumors with heavily pigmented overlying skin. To circumvent these barriers, we use interstitial laser irradiation to induce the desired photothermal effects. The purpose of this study is to analyze the thermal effects of interstitial irradiation using proton resonance frequency (PRF). An 805-nm near-infrared laser with an interstitial cylindrical diffuser was used to treat rat mammary tumors. Different power settings (1.0, 1.25, and 1.5 W) were applied with an irradiation duration of 10 min. The temperature distributions of the treated tumors were measured by a 7 T magnetic resonance imager using PRF. We found that temperature distributions in tissue depended on both laser power and time settings, and that variance in tissue composition has a major influence in temperature elevation. The temperature elevations measured during interstitial laser irradiation by PRF and thermocouple were consistent, with some variations due to tissue composition and the positioning of the thermocouple's needle probes. Our results indicated that, for a tissue irradiation of 10 min, the elevation of rat tumor temperature ranged from 8 to 11°C for 1 W and 8 to 15°C for 1.5 W. This is the first time a 7 T magnetic resonance imager has been used to monitor interstitial laser irradiation via PRF. Our work provides a basic understanding of the photothermal interaction needed to control the thermal damage inside a tumor using interstitial laser treatment. Our work may lead to an optimal protocol for future cancer treatment using interstitial phototherapy in conjunction with immunotherapy.