Evaluation of a large healthy lifestyle program: informing program implementation and scale-up in the prevention of obesity.
The Healthy Lifestyle Program for women (HeLP-her) is a low-intensity, self-management program which has demonstrated efficacy in preventing excess weight gain in women. However, little is known about the implementation, reach, and sustainability of low-intensity prevention programs in rural settings, where risk for obesity in women is higher than urban settings. We aimed to evaluate a low-intensity healthy lifestyle program delivered to women in a rural setting to inform development of effective community prevention programs. A mixed method hybrid implementation and evaluation study, guided by the RE-AIM framework (addressing the Reach, Effectiveness, Adoption, Implementation, and Maintenance), was undertaken. Data collection tools included anthropometric measures, program checklists, questionnaires, and semi-structured interviews with participants and local stakeholders. The RE-AIM self-audit tool was applied to assess evaluation rigor. Six hundred and forty-nine women from 41 relatively socio-economic disadvantaged communities in Australia participated: mean age 39.6 years (±SD 6.7) and body mass index of 28.8 kg/m2 (±SD 6.9). A between-group weight difference of -0.92 kg (95% CI -1.67 to -0.16) showed program effectiveness. Reach was broad across 41 towns with 62% of participants reporting influencing some of the health behaviors of their families. Strong implementation fidelity was achieved with good retention rates at 1 year (76%) and high participant satisfaction (82% of participants willing to recommend this program). Over 300 multi-level community partnerships were established supporting high adoption. Stakeholders reported potential capacity to implement and sustain the prevention program in resource poor rural settings, due to the low-intensity design and minimal resources required. Our comprehensive RE-AIM evaluation demonstrates that an evidence-based obesity prevention program can be successfully implemented in real-world settings. The program achieved broad reach, effectiveness, and satisfaction at the community and stakeholder level, revealing potential for program sustainability. The evaluation addressed implementation knowledge gaps to support future obesity prevention program scale-up. Australian and New Zealand Clinical Trial Registry ACTRN 12612000115831 [ http://www.anzctr.org.au/ ].


Bis(phenolate)amine-supported lanthanide borohydride complexes for styrene and trans-1,4-isoprene (co-)polymerisations.
New bis(phenolate)amine-supported neodymium borohydride complexes and their previously reported samarium analogues were tested as catalysts for the polymerisation of styrene and isoprene. Reaction of Na2O2N(L) (L = py, OMe, NMe2) with Nd(BH4)3(THF)3 afforded the borohydride complexes Nd(O2N(L))(BH4)(THF) (L = py (1-Nd), OMe (2-Nd), NMe2 (3-Nd)). Complex 1-Nd has shown a propensity to form phenolate-O-bridged dimer [Nd(μ-O2N(py))(BH4)]2 (1′-Nd) as previously observed with the samarium analogues Sm(O2N(L))(BH4)(THF) (L = py or Pr). X-ray structures of 1′-Nd and 2-Nd were determined and are presented. The neodymium borohydride complexes 1-Nd to 3-Nd and their samarium analogues Sm(O2N(L))(BH4)(THF)x (L = py (1-Sm), OMe (2-Sm), NMe2 (3-Sm), Pr (4-Sm)) were tested as catalysts for the polymerisation of isoprene and styrene in the presence of n-butylethylmagnesium (Mg((n)Bu)(Et)). All complexes were found to be active for the polymerisation of isoprene in these conditions, leading to polyisoprene up to 95.1% trans-1,4 stereoregular. They were also found to be active for the polymerisation of styrene leading to atactic polystyrene in all cases. Interestingly, samarium-based complexes were found to be more active than the neodymium ones toward this latter monomer, in sharp contrast to what is usually observed with rare earth borohydride complexes. The structure of both trans-polyisoprenes and polystyrenes obtained were studied in detail by MALDI-ToF analysis in order to better understand the polymerisation mechanisms. The coordinative chain transfer polymerisation (CCTP) of both monomers was further conducted using Mg((n)Bu)(Et) as transfer agent. Finally, the statistical copolymerisation of isoprene and styrene was examined using these catalytic systems, leading to the formation of poly[(trans-1,4-isoprene)-co-styrene] with up to 39% of styrene moieties inserted in a highly trans-1,4-stereoregular polyisoprene.


Peripheral endocannabinoid system dysregulation in first-episode psychosis.
Several hypotheses involving alterations of the immune system have been proposed among etiological explanations for psychotic disorders. The endocannabinoid system (ECS) has a homeostatic role as an endogenous neuroprotective and anti-inflammatory system. Alterations of this system have been associated with psychosis. Cannabis use is a robust risk factor for these disorders that could alter the ECS signalling. In this study, 95 patients with a first episode of psychosis (FEP) and 90 healthy controls were recruited. Protein expression of cannabinoid receptor 2 (CB2), the protein levels of the main endocannabinoid synthesizing enzymes N-acyl phosphatidylethanolamine phospholipase (NAPE) and diacylglycerol lipase (DAGL), and of degradation enzymes fatty acid amide hydrolase (FAAH) and monoacylglycerol lipase (MAGL) were determined by western blot analysis in peripheral blood mononuclear cells (PBMCs). Patients with a FEP showed a decreased expression of CB2 and of both endocannabinoids synthesizing enzymes (NAPE and DAGL) in comparison to healthy controls. After controlling for age, gender, body mass index, and cannabis use, NAPE and DAGL expression remained significantly decreased, whereas FAAH and MAGL expression were increased. On the other hand, FEP subjects with history of severe cannabis use showed a larger ECS dysregulation compared with healthy controls. These results indicate an ECS dysregulation in PBMC of FEP patients. The alteration of the ECS presented at the initial phases of psychosis could be contributing to the pathophysiology of the disease and constitutes a possible biomarker of psychotic disorders and an interesting pharmacological target to take into account for therapeutic purposes.


Myxofibrosarcoma presenting in the skin: clinicopathological features and differential diagnosis with cutaneous myxoid neoplasms.
Myxofibrosarcoma (myxoid malignant fibrous histiocytoma) is one of the most common fibroblastic sarcomas in the older patient, where it can sometimes present with anatomically deceptive boundaries. Myxofibrosarcoma is now fully characterized as a distinctive and definable pathologic entity. Clinically there is a tendency for predominantly subcutaneous, multinodular, diffusely infiltrative growth, which may extend to the overlying dermis and present as a cutaneous lesion. Histologically myxofibrosarcoma comprises a spectrum ranging from hypocellular low-grade myxoid to high-grade pleomorphic sarcoma. We report herein 6 cases of myxofibrosarcoma each with dermatological presentation as a cutaneous nodule. The dermal component in each of the lesions was low- to intermediate-grade and predominantly myxoid resulting in confusion with benign myxoid neoplasms in small biopsy specimens. The purpose of this series is to focus the attention of workers in dermatology on a subject rarely discussed in dermatopathology literature: the cutaneous presentation of myxofibrosarcoma and the potential for clinical and histologic misinterpretation, as benign dermal lesions.


Web-based smoking-cessation programs: results of a randomized trial.
Initial trials of web-based smoking-cessation programs have generally been promising. The active components of these programs, however, are not well understood. This study aimed to (1) identify active psychosocial and communication components of a web-based smoking-cessation intervention and (2) examine the impact of increasing the tailoring depth on smoking cessation. Randomized fractional factorial design. Two HMOs: Group Health in Washington State and Henry Ford Health System in Michigan. 1866 smokers. A web-based smoking-cessation program plus nicotine patch. Five components of the intervention were randomized using a fractional factorial design: high- versus low-depth tailored success story, outcome expectation, and efficacy expectation messages; high- versus low-personalized source; and multiple versus single exposure to the intervention components. Primary outcome was 7 day point-prevalence abstinence at the 6-month follow-up. Abstinence was most influenced by high-depth tailored success stories and a high-personalized message source. The cumulative assignment of the three tailoring depth factors also resulted in increasing the rates of 6-month cessation, demonstrating an effect of tailoring depth. The study identified relevant components of smoking-cessation interventions that should be generalizable to other cessation interventions. The study also demonstrated the importance of higher-depth tailoring in smoking-cessation programs. Finally, the use of a novel fractional factorial design allowed efficient examination of the study aims. The rapidly changing interfaces, software, and capabilities of eHealth are likely to require such dynamic experimental approaches to intervention discovery.


A unified understanding of the direct coordination of NO to first-transition-row metal centers in metal-ligand complexes.
The binding of nitric oxide (NO) to heme-proteins is an important biochemical process involved in a variety of physiological functions. Here, using hybrid density-functional calculations, we systematically investigate the adsorption of NO to first-transition-row metal centers in metal-ligand complexes. Through the comparative study for different transition metal (TM) centers, we provide a unified understanding of the microscopic interactions of NO with the TM centers and related chemical trends. We found that as the atomic number of the TM center increases, the binding strength of NO is largely reduced from 207 kJ mol-1 to near zero due to the low d-orbital energies for late TM centers. The intermolecular spin coupling between the localized spins at the TM center and the NO molecule is generally antiferromagnetic, except for the case of Sc. The spin-spin coupling is determined in such a way to avoid the energy penalty associated with the electron occupation in the antibonding states of the NO-bound complex. The adsorption strength of NO is generally larger than of CO because the unpaired electron of NO occupies the associated bonding state.


Clinical features of pedophilia and implications for treatment.
The authors discuss the diagnostic criteria for pedophilia and review the literature on its clinical features, including data on prevalence, gender, age of onset, number of victims, frequency and type of acts, violence, impulsivity, and insight. Findings concerning the characteristics of victims (e.g., sex, age, relationship to the pedophile) and research on pedophilic subtypes-exclusive versus nonexclusive; incestuous versus nonincestuous; heterosexual, homosexual, or bisexual-are reviewed. Studies have shown that pedophiles may share many psychiatric features beyond deviant sexual desire, including high rates of comorbid axis I disorders (affective disorders, substance use disorders, impulse control disorders, other paraphilias) as well as severe axis II psychopathology (especially antisocial and Cluster C personality disorders). The authors present several possible etiological models for pedophilia and conclude that further research is needed concerning the etiological role of a childhood history of sexual abuse as well as the underlying neurobiology of deviant sexual arousal and decreased erotic differentiation. Finally, findings concerning pharmacological and cognitive-behavioral treatments for pedophilia are briefly reviewed. Recidivism, drop-out, and noncompliance are significant problems in the treatment of pedophilia. The authors review predictors of treatment outcome and conclude that pedophilia is extremely difficult to treat and that effective treatment needs to be intensive, long-term, and comprehensive, possibly with lifetime follow-up.


Molecular cloning of two metallothionein-like protein genes with differential expression patterns from sweet potato (Ipomoea batatas) leaves.
Metallothionein (MT) is a group of proteins with low molecular masses and high cysteine contents, and is classified into different types, which in general contains two domains (domain 1 and domain 2) with typical amino acid sequences (Rauser 1999). In this report two full-length cDNAs (Y459 and G14) encoding MT-like proteins were isolated from leaves of sweet potato (Ipomoea batatas). Their open reading frames contained 249 and 195 nucleotides (82 and 64 amino acids) for Y459 and G14, respectively, and exhibited a relatively low amino acid sequence similarity (ca. 25.8%). Gene structure studies showed that Y459 had the conserved domain 1 region of type 2 MT; however, the domain 2 region was not conserved and contained additional amino acids between the CxC and CxC spacing. G14 had conserved domains 1 and 2 of type 4 MT except that the last CxC of domain 2 was changed to RxC. Semi-quantitative RT-PCR showed that Y459 was expressed in significant quantity in roots and stems, but was much less in green leaves. During natural and induced (with dark and ethephon, an ethylene-releasing compound, treatments) leaf senescence, Y459 gene expression was significantly enhanced. In contrast, relatively constant gene expression levels were found for G14 in all tissues or treatments analyzed. In conclusion, the two MT-like protein genes of sweet potato display differential gene structures and gene expression patterns, which may be associated with the diverse roles and functions they play in plant physiology in order to cope with particular developmental and environmental cues.


Investigation of spatial distribution of radiocesium in a paddy field as a potential sink.
Surface soils, under various land uses, were contaminated by radionuclides that were released by the Fukushima Daiichi Nuclear Power Plant accident. Because paddy fields are one of the main land uses in Japan, we investigated the spatial distribution of radiocesium and the influence of irrigation water in a paddy field during cultivation. Soil core samples collected at a paddy field in Fukushima showed that plowing had disturbed the original depth distribution of radiocesium. The horizontal distribution of radiocesium did not show any evidence for significant influence of radiocesium from irrigation water, and its accumulation within the paddy field, since the original amount of radiocesium was much larger than was added into the paddy field by irrigation water. However, it is possible that rainfall significantly increases the loading of radiocesium.


Diagnostic error and clinical reasoning.
There is a growing literature on diagnostic errors. The consensus of this literature is that most errors are cognitive and result from the application of one or more cognitive biases. Such biased reasoning is usually associated with 'System 1' (non-analytic, pattern recognition) thinking. We review this literature and bring in evidence from two other fields: research on clinical reasoning, and research in psychology on 'dual-process' models of thinking. We then synthesise the evidence from these fields exploring possible causes of error and potential solutions. We identify that, in fact, there is very little evidence to associate diagnostic errors with System 1 (non-analytical) reasoning. By contrast, studies of dual processing show that experts are as likely to commit errors when they are attempting to be systematic and analytical. We then examine the effectiveness of various approaches to reducing errors. We point out that educational strategies aimed at explaining cognitive biases are unlikely to succeed because of limited transfer. Conversely, there is an accumulation of evidence that interventions directed at specifically encouraging both analytical and non-analytical reasoning have been shown to result in small, but consistent, improvements in accuracy. Diagnostic errors are not simply a consequence of cognitive biases or over-reliance on one kind of thinking. They result from multiple causes and are associated with both analytical and non-analytical reasoning. Limited evidence suggests that strategies directed at encouraging both kinds of reasoning will lead to limited gains in accuracy.


Assessment of a fluorescence-enhanced optical imaging system using the Hotelling observer.
This study represents a first attempt to assess the detection capability of a fluorescence-enhanced optical imaging system as quantified by the Hotelling observer. The imaging system is simulated by the diffusion approximation of the time-dependent radiative transfer equation, which describes near infra-red (NIR) light propagation through clinically relevant tissue volumes. The random structures in the background are introduced using a lumpy-object model as a representation of anatomical structure as well as non-uniform distribution of disease markers. The systematic errors and noise associated with the actual experimental conditions are incorporated into the simulated boundary measurements to acquire imaging data sets. A large number of imaging data sets are considered in order to perform Hotelling observer studies. We find that the signal-to-noise ratio (SNR) of Hotelling observer (i) decreases as the strength of lumpy perturbations in the background increases, (ii) decreases as the target depth increases, and (iii) increases as excitation light leakage decreases, and reaches a maximum for filter optical density values of 5 or higher.


Congenital pulmonary vascular anomalies.
Congenital pulmonary vascular anomalies are typically found in infancy or early childhood however, some may remain silent and present in adult patients. Anomalies may be separated into anatomic categories based on involvement of the pulmonary arteries, pulmonary veins or both with or without involvement of the lung parenchyma. Association with congenital heart disease and other syndromes is very common. Computed tomography (CT) and magnetic resonance imaging (MRI) are both invaluable at assessment of these anomalies allowing for both diagnosis and detailed treatment planning. This article will focus primarily on the use of CT, as the high resolution evaluation of the lung parenchyma is also important in many of these conditions. In young patients especially, rapid heart rate and concerns of radiation exposure are important considerations when performing CT. This article will discuss scan techniques as well as clinical diagnostic considerations and basic endovascular treatment of congenital pulmonary vascular anomalies.


[Candida albicans isolated from buccal cavity of children with Down's syndrome: occurrence and growth inhibition by Streptomyces sp].
Comparison of the presence of Candida yeasts in the buccal cavity of children without and with Down's syndrome showed a statistically significant difference in the case of children that were affected by this chromosomopathy, rendering them more predisposed to buccal candidiasis, probably due to anatomicophysiologic alterations of the mouth resulting from trisomy of chromosome 21. Constant recurrence of buccal candidiasis in children with this chromosomal alteration lead to the search for a possible therapeutic alternative. Seeking to determine the antifungal activity of Streptomyces sp isolated from various Brazilian soils, 5 strains have been tested for Candida albicans isolated from the buccal cavity of children with Down's syndrome. It was observed that the isolate presented a diversity in the size of the halos (9-31 mm in diameter) of growth inhibition of the yeasts, suggesting a possible use as a therapeutic antifungal.


Young Women With Breast Cancer: Treatment, Care, and Nursing Implications.
Young women with breast cancer (YWBC) are more likely to have aggressive disease, carry mutations for hereditary cancer genes, and experience higher mortality. They also may face reduced fertility because of the toxicity of chemotherapy. This article aims to present a review of YWBC treatments, sequelae of treatment, and psychosocial challenges. The authors performed a review of guideline-supported treatment options, patient resources, and nursing implications. Because of high-risk cancers and a lack of specific treatment guidelines, healthcare providers may consider aggressive treatments for younger patients. However, studies indicate that the foundation for treatment decisions for YWBC are best based on disease stage and National Comprehensive Cancer Network guidelines.


The neutrophilic leukocyte in wound repair a study with antineutrophil serum.
The role of the neutrophilic leukocyte in wound healing was investigated by observing the progress of repair in the absence of these cells. Circulating neutrophils were eliminated in guinea pigs by the administration of antineutrophil serum (ANS) 24 hr before wounding and by daily injections throughout a 10 day period of healing. Control animals received normal rabbit serum at the same dose levels and times. The wounds consisted of six linear incisions in the dorsal skin of the animals.The contents of 24-hr neutropenic and control wounds were compared by quantitating the major cellular and extracellular wound components using a histometric technique. At 24 hr, there were no differences between control and neutropenic wounds in the per cent of total wound volume occupied by mononuclear leukocytes and fibrin. The neutropenic wounds had no neutrophils, had a significantly decreased volume of fluid space, and an increased volume of red cells, as compared with controls. The differences in numbers of erythrocytes and amount of fluid space in these two sets of wounds may be related to substances within neutrophils that promote lysis of erythrocytes or affect vascular permeability. In spite of the lack of neutrophils in the ANS-treated animals during the 10 days of healing, no differences were observed between the control and neutropenic wounds relative to the rate of wound debridement or the extent of repair. The wounds from the two groups of animals were identical in cellularity and degree of connective tissue formation. These observations support the notion that neither wound debridement nor the formation of granulation tissue are dependent upon the presence of neutrophils. A neutrophil response in early wounds is not an essential antecedent to the infiltration of monocytes, as suggested by previous investigations.


Accommodation of insertions in helices: the mutation in hemoglobin Catonsville (Pro 37 alpha-Glu-Thr 38 alpha) generates a 3(10)-->alpha bulge.
Hemoglobin Catonsville is a mutation of human hemoglobin (an alpha 2 beta 2 tetramer) in which a glutamate residue is inserted into the first turn of a highly conserved 3(10) helix (the C helix) of each alpha subunit. In theory, amino acid insertions (or deletions) in protein helices can be accommodated via two distinct mechanisms. One, termed the register shift mechanism, preserves the geometry of the helix while requiring all of the residues on one flank of the insertion site to rotate by 100 degrees in the case of an alpha helix or by 120 degrees in the case of a 3(10) helix. The other, termed the bulge (or indentation) mechanism, distorts the local geometry of the helix but does not alter the helix register. High-resolution X-ray diffraction analysis of deoxyhemoglobin Catonsville shows that the inserted residue is accommodated as a bulge, demonstrating that this is a viable mechanism. (In contrast, no such evidence is yet available for the register shift mechanism.) More specifically, the insertion converts one turn of the C helix from 3(10) geometry to alpha helix-like geometry, raising the possibility that a common mechanism for accommodating insertions and deletions within helices may involve localized interconversions between 3(10), alpha, and pi helical structures.


Molecular origin of high free energy barriers for alkali metal ion transfer through ionic liquid-graphene electrode interfaces.
In this work we study mechanisms of solvent-mediated ion interactions with charged surfaces in ionic liquids by molecular dynamics simulations, in an attempt to reveal the main trends that determine ion-electrode interactions in ionic liquids. We compare the interfacial behaviour of Li(+) and K(+) at a charged graphene sheet in a room temperature ionic liquid, 1-butyl-3-methylimidazolium tetrafluoroborate, and its mixtures with lithium and potassium tetrafluoroborate salts. Our results show that there are dense interfacial solvation structures in these electrolytes that lead to the formation of high free energy barriers for these alkali metal cations between the bulk and direct contact with the negatively charged surface. We show that the stronger solvation of Li(+) in the ionic liquid leads to the formation of significantly higher interfacial free energy barriers for Li(+) than for K(+). The high free energy barriers observed in our simulations can explain the generally high interfacial resistance in electrochemical storage devices that use ionic liquid-based electrolytes. Overcoming these barriers is the rate-limiting step in the interfacial transport of alkali metal ions and, hence, appears to be a major drawback for a generalised application of ionic liquids in electrochemistry. Some plausible strategies for future theoretical and experimental work for tuning them are suggested.


Decision tree algorithms predict the diagnosis and outcome of dengue fever in the early phase of illness.
Dengue is re-emerging throughout the tropical world, causing frequent recurrent epidemics. The initial clinical manifestation of dengue often is confused with other febrile states confounding both clinical management and disease surveillance. Evidence-based triage strategies that identify individuals likely to be in the early stages of dengue illness can direct patient stratification for clinical investigations, management, and virological surveillance. Here we report the identification of algorithms that differentiate dengue from other febrile illnesses in the primary care setting and predict severe disease in adults. A total of 1,200 patients presenting in the first 72 hours of acute febrile illness were recruited and followed up for up to a 4-week period prospectively; 1,012 of these were recruited from Singapore and 188 from Vietnam. Of these, 364 were dengue RT-PCR positive; 173 had dengue fever, 171 had dengue hemorrhagic fever, and 20 had dengue shock syndrome as final diagnosis. Using a C4.5 decision tree classifier for analysis of all clinical, haematological, and virological data, we obtained a diagnostic algorithm that differentiates dengue from non-dengue febrile illness with an accuracy of 84.7%. The algorithm can be used differently in different disease prevalence to yield clinically useful positive and negative predictive values. Furthermore, an algorithm using platelet count, crossover threshold value of a real-time RT-PCR for dengue viral RNA, and presence of pre-existing anti-dengue IgG antibodies in sequential order identified cases with sensitivity and specificity of 78.2% and 80.2%, respectively, that eventually developed thrombocytopenia of 50,000 platelet/mm(3) or less, a level previously shown to be associated with haemorrhage and shock in adults with dengue fever. This study shows a proof-of-concept that decision algorithms using simple clinical and haematological parameters can predict diagnosis and prognosis of dengue disease, a finding that could prove useful in disease management and surveillance.


High-fertility phenotypes: two outbred mouse models exhibit substantially different molecular and physiological strategies warranting improved fertility.
Animal models are valuable tools in fertility research. Worldwide, there are more than 400 transgenic or knockout mouse models available showing a reproductive phenotype; almost all of them exhibit an infertile or at least subfertile phenotype. By contrast, animal models revealing an improved fertility phenotype are barely described. This article summarizes data on two outbred mouse models exhibiting a 'high-fertility' phenotype. These mouse lines were generated via selection over a time period of more than 40 years and 161 generations. During this selection period, the number of offspring per litter and the total birth weight of the entire litter nearly doubled. Concomitantly with the increased fertility phenotype, several endocrine parameters (e.g. serum testosterone concentrations in male animals), physiological parameters (e.g. body weight, accelerated puberty, and life expectancy), and behavioral parameters (e.g. behavior in an open field and endurance fitness on a treadmill) were altered. We demonstrate that the two independently bred high-fertility mouse lines warranted their improved fertility phenotype using different molecular and physiological strategies. The fertility lines display female- as well as male-specific characteristics. These genetically heterogeneous mouse models provide new insights into molecular and cellular mechanisms that enhance fertility. In view of decreasing fertility in men, these models will therefore be a precious information source for human reproductive medicine. Translated abstract A German translation of abstract is freely available at http://www.reproduction-online.org/content/147/4/427/suppl/DC1.


Methods for high-throughput MethylCap-Seq data analysis.
Advances in whole genome profiling have revolutionized the cancer research field, but at the same time have raised new bioinformatics challenges. For next generation sequencing (NGS), these include data storage, computational costs, sequence processing and alignment, delineating appropriate statistical measures, and data visualization. Currently there is a lack of workflows for efficient analysis of large, MethylCap-seq datasets containing multiple sample groups. The NGS application MethylCap-seq involves the in vitro capture of methylated DNA and subsequent analysis of enriched fragments by massively parallel sequencing. The workflow we describe performs MethylCap-seq experimental Quality Control (QC), sequence file processing and alignment, differential methylation analysis of multiple biological groups, hierarchical clustering, assessment of genome-wide methylation patterns, and preparation of files for data visualization. Here, we present a scalable, flexible workflow for MethylCap-seq QC, secondary data analysis, tertiary analysis of multiple experimental groups, and data visualization. We demonstrate the experimental QC procedure with results from a large ovarian cancer study dataset and propose parameters which can identify problematic experiments. Promoter methylation profiling and hierarchical clustering analyses are demonstrated for four groups of acute myeloid leukemia (AML) patients. We propose a Global Methylation Indicator (GMI) function to assess genome-wide changes in methylation patterns between experimental groups. We also show how the workflow facilitates data visualization in a web browser with the application Anno-J. This workflow and its suite of features will assist biologists in conducting methylation profiling projects and facilitate meaningful biological interpretation.


Tuberculosis Treatment Outcome and Drug Resistance in Lambaréné, Gabon: A Prospective Cohort Study.
Despite overall global progress in tuberculosis (TB) control, TB remains one of the deadliest communicable diseases. This study prospectively assessed TB epidemiology in Lambaréné, Gabon, a Central African country ranking 10th in terms of TB incidence rate in the 2014 World Health Organization TB report. In Lambaréné, between 2012 and 2014, 201 adult and pediatric TB patients were enrolled and followed up; 66% had bacteriologically confirmed TB and 95% had pulmonary TB. The human immunodeficiency virus (HIV) coinfection rate was 42% in adults and 16% in children. Mycobacterium tuberculosis and Mycobacterium africanum were identified in 82% and 16% of 108 culture-confirmed TB cases, respectively. Isoniazid (INH) and streptomycin yielded the highest resistance rates (13% and 12%, respectively). The multidrug resistant TB (MDR-TB) rate was 4/91 (4%) and 4/13 (31%) in new and retreatment TB cases, respectively. Treatment success was achieved in 53% of patients. In TB/HIV coinfected patients, mortality rate was 25%. In this setting, TB epidemiology is characterized by a high rate of TB/HIV coinfection and low treatment success rates. MDR-TB is a major public health concern; the need to step-up in-country diagnostic capacity for culture and drug susceptibility testing as well as access to second-line TB drugs urgently requires action.


Fabrication of high aspect ratio metal nanotips by nanosecond pulse laser melting.
The authors have developed an approach to fabricate sharp and high aspect ratio metal tips using nanosecond pulse laser melting. A quartz wafer covered with a thin chromium (Cr) film was placed on top of a second wafer with a sub-micrometer gap between them and the Cr film facing the second wafer. Then an excimer laser pulse (308 nm wavelength, 20 ns pulse duration) was shone from the back of the quartz wafer and melted the Cr film momentarily (several hundred nanoseconds). It is found that the molten Cr films can self-form discrete metal pillars connecting the two wafers. After separating the two wafers, nanotips were formed at the broken pillar necks. The sharpest tip achieved has an apex diameter 10 nm and height 180 nm. The self-formation of Cr pillars between the two wafers was attributed to the attractive electrostatic force caused by the work function difference of two wafers that were in close proximity. This technique could be extended to other metals, and a periodic uniform tip array could be obtained by pre-patterning the metal into identical isolated mesas and precisely controlling the gap between the two wafers.


Association of Longitudinal Changes of Physical Activity on Smoking Cessation Among Young Daily Smokers.
To our knowledge, no longitudinal epidemiological study among daily smokers has examined the effects of physical activity change/ trajectory on smoking cessation. The purpose of this study was to examine the longitudinal effects of changes in physical activity on smoking cessation among a national sample of young (16-24 y) daily smokers. Data from the 2003-2005 National Youth Smoking Cessation Survey were used (N = 1178). Using hierarchical agglomerative cluster analysis, 5 distinct self-reported physical activity trajectories over 3 time periods (baseline, 12-month, and 24-month follow-up) were observed, including stable low physical activity, decreasing physical activity, curvilinear physical activity, stable high physical activity, and increasing physical activity. Nicotine dependence (Heaviness of Smoking Index) and demographic parameters were assessed via survey. With stable low physical activity (16.2% quit smoking) serving as the referent group, those in the stable high physical activity (24.8% quit smoking) group had 1.8 greater odds of not smoking at the 24-month follow-up period (odds ratio = 1.81; 95% confidence interval, 1.12-2.91) after adjusting for nicotine dependence, age, gender, race-ethnicity, and education. Maintenance of regular physical activity among young daily smokers may help to facilitate smoking cessation.


Influence of Pulsed Electric Fields and Mitochondria-Cytoskeleton Interactions on Cell Respiration.
Pulsed electric fields with microsecond pulse width (μsPEFs) are used clinically; namely, irreversible electroporation/Nanoknife is used for soft tissue tumor ablation. The μsPEF pulse parameters used in irreversible electroporation (0.5-1 kV/cm, 80-100 pulses, ∼100 μs each, 1 Hz frequency) may cause an internal field to develop within the cell because of the disruption of the outer cell membrane and subsequent penetration of the electric field. An internal field may disrupt voltage-sensitive mitochondria, although the research literature has been relatively unclear regarding whether such disruptions occur with μsPEFs. This investigation reports the influence of clinically used μsPEF parameters on mitochondrial respiration in live cells. Using a high-throughput Agilent Seahorse machine, it was observed that μsPEF exposure comprising 80 pulses with amplitudes of 600 or 700 V/cm did not alter mitochondrial respiration in 4T1 cells measured after overnight postexposure recovery. To record alterations in mitochondrial function immediately after μsPEF exposure, high-resolution respirometry was used to measure the electron transport chain state via responses to glutamate-malate and ADP and mitochondrial membrane potential via response to carbonyl cyanide-p-trifluoromethoxyphenylhydrazone. In addition to measuring immediate mitochondrial responses to μsPEF exposure, measurements were also made on cells permeabilized using digitonin and those with compromised cytoskeleton due to actin depolymerization via treatment with the drug latrunculin B. The former treatment was used as a control to tease out the effects of plasma membrane permeabilization, whereas the latter was used to investigate indirect effects on the mitochondria that may occur if μsPEFs impact the cytoskeleton on which the mitochondria are anchored. Based on the results, it was concluded that within the pulse parameters tested, μsPEFs alone do not hinder mitochondrial physiology but can be used to impact the mitochondria upon compromising the actin. Mitochondrial susceptibility to μsPEF after actin depolymerization provides, to our knowledge, a novel avenue for cancer therapeutics.


pH-jump studies at subzero temperatures on an intermediate in the reaction of xanthine oxidase with xanthine.
Xanthine oxidase is stable and active in aqueous dimethyl sulphoxide solutions of up to at least 57% (w/w). Simple techniques are described for mixing the enzyme in this solvent at--82 degrees C, with its substrate, xanthine. When working at high pH values under such conditions, no reaction occurred, as judged by the absence of e.p.r. signals. On warming to--60 degrees C, for 10 min, however, the Very Rapid molybdenum(V) e.p.r. signal was obtained. This signal did not change on decreasing the pH, while maintaining the sample in liquid nitrate reductase, caused its molybdenum(V) e.p.r. signal to change from the high-pH to the low-pH form. These findings are not compatible with the conclusions of Edmondson, Ballou, Van Heuvelen, Palmer & Massey [J. Biol. Chem. (1973) 248, 6135-6144], that the Very Rapid signal is in prototropic equilibrium with the Rapid signal, and should be important in understanding the mechanism of action of the enzyme. They emphasize the unique nature of the intermediate represented by the Very Rapid e.p.r. signal. The possible value of the pK for loss of an exchangeable proton from the Rapid signal is discussed.


Transmission of Onchocerca volvulus continues in Nyagak-Bondo focus of northwestern Uganda after 18 years of a single dose of annual treatment with ivermectin.
The objective of the study was to determine whether annual ivermectin treatment in the Nyagak-Bondo onchocerciasis focus could safely be withdrawn. Baseline skin snip microfilariae (mf) and nodule prevalence data from six communities were compared with data collected in the 2011 follow-up in seven communities. Follow-up mf data in 607 adults and 145 children were compared with baseline (300 adults and 58 children). Flies collected in 2011 were dissected, and poolscreen analysis was applied to ascertain transmission. Nodule prevalence in adults dropped from 81.7% to 11.0% (P < 0.0001), and mf prevalence dropped from 97.0% to 23.2% (P < 0.0001). In children, mf prevalence decreased from 79.3% to 14.1% (P < 0.0001). Parous and infection rates of 401 flies that were dissected were 52.9% and 1.5%, respectively, whereas the infective rate on flies examination by polymerase chain reaction (PCR) was 1.92% and annual transmission potential was 26.9. Stopping ivermectin treatment may result in onchocerciasis recrudescence.


Bio-functional constituents from the stems of Liriodendron tulipifera.
Four known compounds have been isolated from the stems of Liriodendron tulipifera, and the structures of these pure constituents were determined using spectroscopic analysis. Isolated compounds were screened for free radical scavenging ability, metal chelating power assay and ferric reducing antioxidant power assay (FRAP). The anti-tyrosinase effects of L. tulipifera compounds were calculated the inhibition of hydroxylation of L-tyrosine to L-dopa according to an in vitro mushroom tyrosinase assay. The study also examined the bio-effects of the four compounds on the human melanoma A375.S2, and showed that liriodenine (1) and (-)-norglaucine (4) significantly inhibited the proliferation of melanoma cells in the cell viability assay. Wound healing results indicated that liriodenine (1), (-)-glaucine (3) and (-)-norglaucine (4) exerted anti-migration potential. Interestingly, (-)-glaucine (3), neither liriodenine (1) nor (-)-norglaucine (4) showed promising anti-migration potential without inducing significant cytotoxicity. Furthermore, a dramatically increased level of intracellular reactive oxygen species (ROS) was detected from (-)-glaucine (3). The cell cycle assessment demonstrated a moderate G2/M accumulation by (-)-glaucine (3). The above results revealed the anti-cancer effects of L. tulipifera compounds, especially on the anti-migration ability indicating the promising chemopreventive agents to human skin melanoma cells.


Renal changes and acute kidney injury in covid-19: a systematic review.
We aimed to present a review of renal changes in patients with COVID-19. We performed a systematic review of the literature to identify original articles regarding clinical, laboratory, and anatomopathological kidney changes in patients infected with SARS-CoV-2 published until May 7, 2020. The search was carried out across PubMed, Scopus, and Embase using the keywords "COVID-19", "coronavirus", "SARS-CoV-2", "kidney injury" and "kidney disease". Fifteen studies presented clinical and laboratory renal changes in patients with COVID-19, and three addressed anatomopathological changes. Acute kidney injury (AKI) was a relevant finding in patients with COVID-19. There were also significant changes in laboratory tests that indicated kidney injury, such as increased serum creatinine and blood urea nitrogen (BUN), proteinuria, and hematuria. The presence of laboratory abnormalities and AKI were significant in severely ill patients. There was a considerable prevalence of AKI among groups of patients who died of COVID-19. Histopathological analysis of the kidney tissue of patients infected with SARS-CoV-2 suggested that the virus may directly affect the kidneys. Although COVID-19 affects mainly the lungs, it can also impact the kidneys. Increased serum creatinine and BUN, hematuria, proteinuria, and AKI were frequent findings in patients with severe COVID-19 and were related to an increased mortality rate. Further studies focusing on renal changes and their implications for the clinical condition of patients infected with the novel coronavirus are needed.


Muscular and centre of pressure response to sudden release of load in symmetric and asymmetric stoop lifting tasks.
Sudden changes in load during asymmetric lifting may be associated with a particularly high risk of loss of balance and spinal injury. Centre of pressure (COP) motions and electromyographic responses of trunk and lower limb muscles were studied in 10 normal male volunteers during sudden release of 20, 40, 60 and 80N stoop lifting loads in symmetric and asymmetric postures. Similar overall COP responses and muscular response strategies to sudden release of load were seen in both postures, although the asymmetric posture showed a larger medio-lateral COP displacements and greater co-contraction asymmetries. While sudden release of load in asymmetric stoop lifting does not seem to involve a greater risk of fall than symmetric lifting, the muscular response results in more complex and asymmetric loading of the trunk, indicating greater localised segmental loading and therefore increased risk of tissue injury.


Stimulated bacterial growth under elevated p CO₂: results from an off-shore mesocosm study.
Marine bacteria are the main consumers of freshly produced organic matter. Many enzymatic processes involved in the bacterial digestion of organic compounds were shown to be pH sensitive in previous studies. Due to the continuous rise in atmospheric CO2 concentration, seawater pH is presently decreasing at a rate unprecedented during the last 300 million years but the consequences for microbial physiology, organic matter cycling and marine biogeochemistry are still unresolved. We studied the effects of elevated seawater pCO2 on a natural plankton community during a large-scale mesocosm study in a Norwegian fjord. Nine Kiel Off-Shore Mesocosms for Future Ocean Simulations (KOSMOS) were adjusted to different pCO2 levels ranging initially from ca. 280 to 3000 µatm and sampled every second day for 34 days. The first phytoplankton bloom developed around day 5. On day 14, inorganic nutrients were added to the enclosed, nutrient-poor waters to stimulate a second phytoplankton bloom, which occurred around day 20. Our results indicate that marine bacteria benefit directly and indirectly from decreasing seawater pH. During the first phytoplankton bloom, 5-10% more transparent exopolymer particles were formed in the high pCO2 mesocosms. Simultaneously, the efficiency of the protein-degrading enzyme leucine aminopeptidase increased with decreasing pH resulting in up to three times higher values in the highest pCO2/lowest pH mesocosm compared to the controls. In general, total and cell-specific aminopeptidase activities were elevated under low pH conditions. The combination of enhanced enzymatic hydrolysis of organic matter and increased availability of gel particles as substrate supported up to 28% higher bacterial abundance in the high pCO2 treatments. We conclude that ocean acidification has the potential to stimulate the bacterial community and facilitate the microbial recycling of freshly produced organic matter, thus strengthening the role of the microbial loop in the surface ocean.


The North American opioid epidemic: current challenges and a call for treatment as prevention.
There is a need for creative, public health-oriented solutions to the increasingly intractable problems associated with the North American opioid epidemic. This epidemic is a fundamentally continental problem, as routes of migration, drug demand, and drug exchange link the USA with Mexico and Canada. The challenges faced throughout North America include entrenched prescribing practices of opioid medications, high costs and low availability of medication-assisted treatment (MAT), and policy approaches that present substantial barriers to care.We advocate for the scale up of a low-threshold treatment model for MAT that incorporates the best practices in addiction treatment. Such a model would remove barriers to care through widespread treatment availability and affordability and also a policy of decriminalization. Given that MAT reduces the frequency of drug injecting among opioid injectors, this treatment model should also be guided by an understanding of the socially communicable nature of injection drug use, such that increasing MAT availability may also prevent the spread of injecting practices to individuals at risk of transitions from non-injection to injection drug use. To that end, the "Treatment as Prevention" model employed to respond to the individual- and population-level risks for HIV/AIDS prevention could be adapted to efforts to halt the North American opioid epidemic.


Influence of autonomic neuropathy of different severities on the hypercapnic drive to breathing in diabetic patients.
To investigate the effects of the autonomic nervous system on control of breathing, the neuromuscular (mouth occlusion pressure at 0.1 s after onset of inspiration [P0.1]) and ventilatory (minute ventilation [VE]) response to progressive hyperoxic hypercapnia was assessed in diabetic patients with autonomic dysfunction of different severity. Eighteen diabetics with autonomic neuropathy, nine with parasympathetic damage (DANp), and nine with parasympathetic and sympathetic damage (DANp+s), as indicated by marked postural hypotension, low increment of diastolic BP during sustained handgrip, and lowest resting catecholamine plasma levels, were studied together with a group of 10 diabetic patients without autonomic neuropathy (D) and a group of 10 normal subjects (C). All subjects had pulmonary function tests, including maximal voluntary ventilation and diffusion of carbon monoxide, measurements of respiratory muscle strength as maximal inspiratory mouth pressure (MIP) and maximal expiratory mouth pressure (MEP), and a CO2 rebreathing test (Read's method). Although in the normal range, lung volumes and FEV1 and forced expiratory flows were lower in the DANp and DANp+s groups than in the D and C groups, MIP and MEP were similar among C and diabetic groups, as well as resting P0.1, VE, tidal volume (VT), and respiratory rate (RR). The slope of the linear relationship between P0.1 and end-tidal PCO2 (PETCO2) was higher in DANp+s (0.63+/-0.07 cm H2O/mm Hg) than in C (0.45+/-0.06 cm H2O/mm Hg; p<0.05) and three times greater in DANp+s than in D (0.26+/-0.03 cm H2O/mm Hg; p<0.001) and DANp (0.24+/-0.03 cm H2O/mm Hg; p<0.001), who in turn showed a lower deltaP0.1/deltaPETCO2 than C. The VE increase with increasing PETCO2 was greater in DANp+s (3.70+/-0.85 L/min/mm Hg) than in DANp (2.13+/-0.20 L/min/mm Hg; p<0.05) and D (2.37+/-0.40 L/min/mm Hg; p=0.07), but not significantly higher from that of C (3.17+/-0.36 L/min/mm Hg). No differences were found for deltaVT/deltaPETCO2 among the groups, whereas the deltaRR/deltaPETCO2 relationship was steeper in DANp+s than in DANp (p<0.05) and D (p=0.055). These data reflect a depressed CO2 response both in D and DANp. The presumable decrease of the sympathetic nerve traffic in DANp+s appears to reverse this abnormality. DANp+s, however, exhibit an enhanced CO2 neuromuscular response even in respect to C, suggesting that the sympathetic nervous system might modulate the output of the respiratory centers to hypercapnic stimulus.


Benefit and cost curves for typical pollination mutualisms.
Mutualisms provide benefits to interacting species, but they also involve costs. If costs come to exceed benefits as population density or the frequency of encounters between species increases, the interaction will no longer be mutualistic. Thus curves that represent benefits and costs as functions of interaction frequency are important tools for predicting when a mutualism will tip over into antagonism. Currently, most of what we know about benefit and cost curves in pollination mutualisms comes from highly specialized pollinating seed-consumer mutualisms, such as the yucca moth-yucca interaction. There, benefits to female reproduction saturate as the number of visits to a flower increases (because the amount of pollen needed to fertilize all the flower's ovules is finite), but costs continue to increase (because pollinator offspring consume developing seeds), leading to a peak in seed production at an intermediate number of visits. But for most plant-pollinator mutualisms, costs to the plant are more subtle than consumption of seeds, and how such costs scale with interaction frequency remains largely unknown. Here, we present reasonable benefit and cost curves that are appropriate for typical pollinator-plant interactions, and we show how they can result in a wide diversity of relationships between net benefit (benefit minus cost) and interaction frequency. We then use maximum-likelihood methods to fit net-benefit curves to measures of female reproductive success for three typical pollination mutualisms from two continents, and for each system we chose the most parsimonious model using information-criterion statistics. We discuss the implications of the shape of the net-benefit curve for the ecology and evolution of plant-pollinator mutualisms, as well as the challenges that lie ahead for disentangling the underlying benefit and cost curves for typical pollination mutualisms.


A 5-year evaluation of the emergency contraception enhanced community pharmacy service provided in Wales.
Access to emergency contraception (EC) has been a core component of attempts to address high teenage pregnancy rates in Wales. A national service was commissioned in 2011, allowing supply of EC free of charge from community pharmacies (CPs). This study investigated 5 years of the EC service, to describe its use and investigate changes in the pattern of use over time. Secondary analyses of data from all National Health Service funded CP EC consultations in Wales between 1 August 2012 and 31 July 2017 (n=181 359). Data comprised standardised clinical and demographic information, in the form of predefined service user responses, submitted for reimbursement by CPs. Overall service provision remained relatively consistent over the study period, with women aged between 13 and 59 years accessing the service. An association was observed between the time since unprotected sexual intercourse and the day on which the service was accessed (Χ2(18)=16 292.327, p<0.001). Almost half (47.9%) of requests were because no contraception had been used, with a strong and positive association for teenagers and women aged 40+ years. A statistically significant and increasing percentage of consultations were accompanied by further sexual health advice (r=0.7, p<0.01). Access to EC through CPs is contributing to reducing teenage conceptions and termination rates. However, action is needed to increase contraception use in all age groups. Reduced availability of CP services on Sundays is a barrier to timely EC access. Findings support an expanded role for community pharmacists in provision of regular contraception.


A Sociotechnical Approach to Evaluating the Impact of ICT on Clinical Care Environments.
Process-supporting information technology holds the potential to increase efficiency, reduce errors, and alter professional roles and responsibilities in a manner which allows improvement in the delivery of patient care. However, clashes between the model of health care work inscribed in these tools with the actual nature of work has resulted in staff resistance and decreased organisational uptake of ICT, as well as the facilitation of unexpected and negative effects in efficiency and patient safety. Sociotechnical theory provides a paradigm against which workflow and transfusion of ICT in healthcare could be better explored and understood. This paper will conceptualise a formative, multi-method longitudinal evaluation process to explore the impact of ICT with an appreciation of the relationship between the social and technical systems within a clinical department. Departmental culture, including clinical work processes and communication patterns will be thoroughly explored before system implementation using both quantitative and qualitative research methods. Findings will be compared with post implementation data, which will incorporate measurement of safety and workflow efficiency indicators. Sociotechnical theory provides a paradigm against which workflow and transfusion of ICT in healthcare could be better explored and understood. However, sociotechnical and multimethod approaches to evaluation do not exist without criticism. Inherent in the protocol are limitations of sociotechnical theory and criticism of the multimethod approach; testing of the methodology in real clinical settings will serve to verify efficacy and refine the process.


Functional Exercise Training and Undulating Periodization Enhances the Effect of Whole-Body Electromyostimulation Training on Running Performance.
The popularity of whole-body electromyostimulation is growing during the last years, but there is a shortage of studies that evaluate its effects on physical fitness and sport performance. In this study, we compared the effects of a periodized and functional whole-body-electromyostimulation training on maximum oxygen uptake (VO2max), ventilatory thresholds (VT1 and VT2), running economy (RE), and lower-body muscle strength in runners, vs. a traditional whole-body-electromyostimulation training. A total of 12 male recreational runners, who had been running 2-3 times per week (90-180 min/week) for at least the previous year and had no previous experience on WB-EMS training, were enrolled in the current study. They were randomly assigned to a periodized and functional whole-body-electromyostimulation training group (PFG) (n = 6; 27.0 ± 7.5 years; 70.1 ± 11.1 kg; 1.75 ± 0.05 m) whose training program involved several specific exercises for runners, or a traditional whole-body-electromyostimulation training group (TG) (n = 6; 25.8 ± 7.4 years; 73.8 ± 9.8 kg; 1.73 ± 0.07 m), whose sessions were characterized by circuit training with 10 dynamic and general exercises without external load. The training programs consisted of one whole-body electromyostimulation session and one 20-min running session per week, during 6 weeks. The PFG followed an undulating periodization model and a selection of functional exercises, whereas the TG followed a traditional session structure used in previous studies. Both groups were instructed to stop their habitual running training program. VO2max, VT1, VT2, RE, and lower body muscle strength (vertical jump) were measured before and after the intervention. The PFG obtained significantly higher improvements when compared with the TG in terms of VO2max (2.75 ± 0.89 vs. 1.03 ± 1.01 ml/kg/min, P = 0.011), VT2 (2.95 ± 1.45 vs. 0.35 ± 0.85 ml/kg/min, P = 0.005), VO2max percentage at VT2 (5.13 ± 2.41 vs. 0.63 ± 1.61%), RE at VT1 (-7.70 ± 2.86 vs. -3.50 ± 2.16 ml/kg/km, P = 0.048), RE at 90% of VT2 (-15.38 ± 4.73 vs. -3.38 ± 4.11 ml/kg/km, P = 0.005), and vertical jump in Abalakov modality (2.95 ± 0.94 vs. 0.52 ± 1.49 cm, P = 0.008). Therefore, we conclude that running performance improvements were better after a 6-week program following an undulating periodization and consisting on functional exercises when compared with a 6-week traditional WB-EMS program.


Understanding Selenium and Glutathione as Antiviral Factors in COVID-19: Does the Viral Mpro Protease Target Host Selenoproteins and Glutathione Synthesis?
Glutathione peroxidases (GPX), a family of antioxidant selenoenzymes, functionally link selenium and glutathione, which both show correlations with clinical outcomes in COVID-19. Thus, it is highly significant that cytosolic GPX1 has been shown to interact with an inactive C145A mutant of Mpro, the main cysteine protease of SARS-CoV-2, but not with catalytically active wild-type Mpro. This seemingly anomalous result is what might be expected if GPX1 is a substrate for the active protease, leading to its fragmentation. We show that the GPX1 active site sequence is substantially similar to a known Mpro cleavage site, and is identified as a potential cysteine protease site by the Procleave algorithm. Proteolytic knockdown of GPX1 is highly consistent with previously documented effects of recombinant SARS-CoV Mpro in transfected cells, including increased reactive oxygen species and NF-κB activation. Because NF-κB in turn activates many pro-inflammatory cytokines, this mechanism could contribute to increased inflammation and cytokine storms observed in COVID-19. Using web-based protease cleavage site prediction tools, we show that Mpro may be targeting not only GPX1, but several other selenoproteins including SELENOF and thioredoxin reductase 1, as well as glutamate-cysteine ligase, the rate-limiting enzyme for glutathione synthesis. This hypothesized proteolytic knockdown of components of both the thioredoxin and glutaredoxin systems is consistent with a viral strategy to inhibit DNA synthesis, to increase the pool of ribonucleotides for RNA synthesis, thereby enhancing virion production. The resulting "collateral damage" of increased oxidative stress and inflammation would be exacerbated by dietary deficiencies of selenium and glutathione precursors.


Inhibition of receptor/G protein coupling by suramin analogues.
Suramin analogues act as direct antagonists of heterotrimeric G proteins because they block the rate-limiting step of G protein activation (i.e., the dissociation of GDP prebound to the G protein alpha subunit). We have used the human brain A1 adenosine receptor and the rat striatal D2 dopamine receptor, two prototypical Gi/G(o)-coupled receptors, as a model system to test whether the following analogues suppress the receptor-dependent activation of G proteins: 8-(3-nitrobenzamido)-1,3,5-naphthalenetrisulfonic acid (NF007), 8-(3-(3-nitrobenzamido)-benzamido)-1,3,5-naphthalenetrisulfonic acid (NF018); 8,8'-(carbonylbis(imino-3,1-phenylene))bis-(1,3,5-naphthalenetr isulfonic acid) (NF023); 8,8'-(carbonylbis(imino-3,1-phenylene)carbonylimino-(3,1- phenylene)) bis(1,3,5-naphthalenetrisulfonic acid) (NF037); and suramin. Suramin and its analogues inhibit the formation of the agonist-specific ternary complex (agonist/receptor/G protein). This inhibition is (i) quasicompetitive with respect to agonist binding in that it can be overcome by increasing receptor occupancy but (ii) does not result from an interaction of the analogues with the ligand binding pocket of the receptors because the binding of antagonists or of agonists in the absence of functional receptor/G protein interaction is not affected. In addition to suppressing the spontaneous release of GDP from defined G protein alpha subunits, suramin and its analogues reduce receptor-catalyzed guanine nucleotide exchange. The site, to which suramin analogues bind, overlaps with the docking site for the receptor on the G protein alpha subunit. The structure-activity relationships for inhibition of agonist binding to the A1 adenosine receptor (suramin > NF037 > NF023) and of agonist binding to the inhibition D2 dopamine receptor (suramin = NF037 > NF023 > NF018) differ. Thus, NF037 discriminates between the ternary complexes formed by the agonist-liganded D2 dopamine receptors and those formed by the A1 adenosine receptor with > 10-fold selectivity. Therefore, our results also show that inhibitors can be identified that selectively uncouple specific receptor/G protein tandems.


Apoptotic process in the monkey small intestinal epithelium: I. Association with glutathione level and its efflux.
The apoptotic process in the normal gastrointestinal mucosa is of interest due to its possible role in physiological cell renewal. The aim of this study was to identify the apoptotic process in the monkey small intestine and the association of glutathione level and its efflux in this process. Monkey small intestinal epithelial cells were separated in to different fractions consisting of villus, middle and crypt cells. Apoptosis was identified by DNA ladder pattern and Hoechst staining. The level of glutathione, its efflux and the enzymes involved in its metabolism were quantitated in these fractions. Apoptotic cells were identified predominantly in the villus tip cell fractions by both DNA ladder pattern and Hoechst dye staining. Glutathione level was 7 fold higher in the crypt cells as compared to villus tip cells the middle cells showing a gradual decrease. A similar pattern was seen in mitochondrial content of glutathione. As the cells mature from crypt to villus, there is increased efflux of GSH, which may be responsible for the decreased level of GSH in apoptotic villus cells. In the monkey small intestine, apoptotic cells are seen in the villus tip fractions and the glutathione level and its efflux may play a role in this process.


Single-cell gene expression analysis reveals genetic associations masked in whole-tissue experiments.
Gene expression in multiple individual cells from a tissue or culture sample varies according to cell-cycle, genetic, epigenetic and stochastic differences between the cells. However, single-cell differences have been largely neglected in the analysis of the functional consequences of genetic variation. Here we measure the expression of 92 genes affected by Wnt signaling in 1,440 single cells from 15 individuals to associate single-nucleotide polymorphisms (SNPs) with gene-expression phenotypes, while accounting for stochastic and cell-cycle differences between cells. We provide evidence that many heritable variations in gene function--such as burst size, burst frequency, cell cycle-specific expression and expression correlation/noise between cells--are masked when expression is averaged over many cells. Our results demonstrate how single-cell analyses provide insights into the mechanistic and network effects of genetic variability, with improved statistical power to model these effects on gene expression.


Antigenic composition of single nano-sized extracellular blood vesicles.
Extracellular vesicles (EVs) are important in normal physiology and are altered in various pathologies. EVs produced by different cells are antigenically different. Since the majority of EVs are too small for routine flow cytometry, EV composition is studied predominantly in bulk, thus not addressing their antigenic heterogeneity. Here, we describe a nanoparticle-based technique for analyzing antigens on single nano-sized EVs. The technique consists of immuno-capturing of EVs with 15-nm magnetic nanoparticles, staining captured EVs with antibodies against their antigens, and separating them from unbound EVs and free antibodies in a magnetic field, followed by flow analysis. This technique allows us to characterize EVs populations according to their antigenic distribution, including minor EV fractions. We demonstrated that the individual blood EVs carry different sets of antigens, none being ubiquitous, and quantified their distribution. The physiological significance of antigenically different EVs and their correlation with different pathologies can now be directly addressed. From the clinical editor: This study reports a nanoparticle-based technique for analyzing antigens on single nano-sized extracellular vehicles (EV). The technique consists of immuno-capturing of EVs with 15-nm magnetic nanoparticles, followed by staining the captured EVs with antibodies and separating them via a magnetic field, followed by flow analysis. This technique enables studies of antigenic properties of individual EVs that conventionally can only be studied in bulk.


Facilitation of motor excitability during listening to spoken sentences is not modulated by noise or semantic coherence.
Comprehending speech can be particularly challenging in a noisy environment and in the absence of semantic context. It has been proposed that the articulatory motor system would be recruited especially in difficult listening conditions. However, it remains unknown how signal-to-noise ratio (SNR) and semantic context affect the recruitment of the articulatory motor system when listening to continuous speech. The aim of the present study was to address the hypothesis that involvement of the articulatory motor cortex increases when the intelligibility and clarity of the spoken sentences decreases, because of noise and the lack of semantic context. We applied Transcranial Magnetic Stimulation (TMS) to the lip and hand representations in the primary motor cortex and measured motor evoked potentials from the lip and hand muscles, respectively, to evaluate motor excitability when young adults listened to sentences. In Experiment 1, we found that the excitability of the lip motor cortex was facilitated during listening to both semantically anomalous and coherent sentences in noise relative to non-speech baselines, but neither SNR nor semantic context modulated the facilitation. In Experiment 2, we replicated these findings and found no difference in the excitability of the lip motor cortex between sentences in noise and clear sentences without noise. Thus, our results show that the articulatory motor cortex is involved in speech processing even in optimal and ecologically valid listening conditions and that its involvement is not modulated by the intelligibility and clarity of speech.


Novel targets for mitochondrial medicine.
Mitochondria-classically viewed as the powerhouses of the cell-have taken center stage in disease pathogenesis and resolution. Mitochondrial dysfunction, which originates from primary defects within the organelle or is induced by environmental stresses, plays a critical role in human disease. Despite their central role in human health and disease, there are no approved drugs that directly target mitochondria. We present possible new druggable targets in mitochondrial biology, including protein modification, calcium ion (Ca(2+)) transport, and dynamics, as we move into a new era of mitochondrial medicine.


Presence of qacEΔ1 and cepA genes and susceptibility to a hospital biocide in clinical isolates of Klebsiella pneumoniae in Iran.
The aim of this study was to evaluate the susceptibility of Klebsiella pneumoniae clinical isolates to antibiotics and to a quaternary ammonium compound (QAC) disinfectant as the concentrations used clinically and to determine the presence of the qacEΔ1 and cepA genes for the first time in Iran. In total, 85 K. pneumoniae isolates were randomly collected from hospitalized patients at the general hospitals in Lorestan, Iran. Antibiotic and antiseptic susceptibility testing was performed according to Clinical and Laboratory Standards Institute recommendations. K. pneumonia isolates were screened by PCR amplification of qacEΔ1 and cepA genes using specific primers and sequence analysis of the amplified regions were also performed. From 85 isolates of K. pneumoniae, 34 (40%) isolates were multidrug resistance (MDR). The evaluation of the susceptibility to the QAC disinfectant revealed that 51 (60%) isolates had reduced susceptibility to QAC disinfectant. The qacEΔ1 gene was detected in 26 isolates (30.6%). While cepA gene was found in 19 isolates (22.3%) of K. pneumonia. Seventy-three percent (19/26) qacEΔ1-positive isolates were detected in the biocide-resistant isolates. Whereas, 63.1% (12/19) cepA-positive isolates were found in the biocide-resistant isolates. Out of qacEΔ1 and cepA-positive isolates, 65.4% (17/26) and 42.1% (8/19) were among MDR isolates, respectively. No significant association of biocide resistance with the presence of qacEΔ1 and cepA genes was observed (P>0.05). The results of present study shows that there was a close link between qacEΔ1 gene and antibiotic resistance, but no significant association of biocide resistance with the presence of qacEΔ1 and cepA genes was observed in K. pneumoniae in Iran.


Cytokinin-dependent regulatory module underlies the maintenance of zinc nutrition in rice.
Zinc (Zn) deficiency is a critical problem in human nutrition. Rice is the main source of calories for nearly half the world's population but has the shortcoming, from a nutritional perspective, of being low in Zn and other essential nutrients. Here we performed analyses with cytokinin-related mutants and transgenic lines to provide unequivocal evidence that cytokinins have a key role in controlling Zn status in plants. Transporters responsible for Zn uptake and chelators for the internal transport of Zn were strictly controlled by cytokinins. Moreover, cytokinin metabolism was regulated in a highly dynamic way in response to Zn status, which allows rice to adapt to heterogeneous Zn availability. Subsequently, fine-tuning of cytokinin metabolism by root-specific expression of a cytokinin degradation enzyme was able to improve both Zn nutrient and yield traits. Importantly, X-ray fluorescence imaging revealed that the increased Zn was broadly distributed from the aleurone layer to the inner endosperm. These findings show that metabolic control of cytokinin could provide the key to breeding Zn-enriched rice.


Positive consequences of surviving a stroke.
This exploratory study focused on gaining insight into the psychological experience of stroke survivors undergoing inpatient rehabilitation. Specifically it investigated whether stroke survivors undergoing inpatient rehabilitation were able to identify any positive consequence of their stroke. This phenomenon, also know as benefit finding or positive reframing, has recently received more attention in the coping literature examining reactions to severe illness. Using a case series methodology and qualitative analytic techniques, 16 stroke survivors were interviewed using standardized open-ended questions related to identification of positive consequences attributed to surviving a stroke. Sixty-three percent of the patients interviewed were able to identify positive consequences of their stroke. The following five themes regarding positive consequences of stroke emerged from this case series: increased social relationships, increased health awareness, change in religious life, personal growth, and altruism. It was concluded that some individuals who have survived an acute stroke are able to reframe their experience in a positive light. This study provides occupational therapists and others with further insight into the coping mechanisms of those individuals who have survived a stroke. The need for further research is stressed.


Evidence of Impaired Proprioception in Chronic, Idiopathic Neck Pain: Systematic Review and Meta-Analysis.
Despite common use of proprioceptive retraining interventions in people with chronic, idiopathic neck pain, evidence that proprioceptive dysfunction exists in this population is lacking. Determining whether proprioceptive dysfunction exists in people with chronic neck pain has clear implications for treatment prescription. The aim of this study was to synthesize and critically appraise all evidence evaluating proprioceptive dysfunction in people with chronic, idiopathic neck pain by completing a systematic review and meta-analysis. MEDLINE, CINAHL, PubMed, Allied and Complementary Medicine, EMBASE, Academic Search Premier, Scopus, Physiotherapy Evidence Database (PEDro), and Cochrane Collaboration databases were searched. All published studies that compared neck proprioception (joint position sense) between a chronic, idiopathic neck pain sample and asymptomatic controls were included. Two independent reviewers extracted relevant population and proprioception data and assessed methodological quality using a modified Strengthening the Reporting of Observational Studies in Epidemiology (STROBE) statement. Thirteen studies were included in the present review. Meta-analysis on 10 studies demonstrated that people with chronic neck pain perform significantly worse on head-to-neutral repositioning tests, with a moderate standardized mean difference of 0.44 (95% confidence interval=0.25, 0.63). Two studies evaluated head repositioning using trunk movement (no active head movement thus hypothesized to remove vestibular input) and showed conflicting results. Three studies evaluated complex or postural repositioning tests; postural repositioning was no different between groups, and complex movement tests were impaired only in participants with chronic neck pain if error was continuously evaluated throughout the movement. A paucity of studies evaluating complex or postural repositioning tests does not permit any solid conclusions about them. People with chronic, idiopathic neck pain are worse than asymptomatic controls at head-to-neutral repositioning tests.


Clinical Outcome of Total Knee Arthroplasty Performed Using Patient-Specific Cutting Guides.
BACKGROUND Patient-specific instruments have recently gained attention for ensuring the accuracy of osteotomy and correct alignment in total knee arthroplasty (TKA). We aimed to report our experience with TKA performed by using CT-based 3D model of cutting guides in our clinic. MATERIAL AND METHODS A total of 36 patients (mean age, 67.58±8.46 years; 24 females, 12 males) who underwent TKA with 3D patient-specific cutting guides in our clinic were included in the study. Differences between preoperatively planned bone resections and actual bone resections were calculated. Femoral and tibial component angles were measured on radiographs of the knee. The change in knee pain was evaluated by patients on a 10-point visual analog scale (VAS). Patients were followed up for 6 months postoperatively. RESULTS The actual bone resection was 0.5-1.5 mm higher than the planned resection for all sides, being statistically significant for posterior lateral and distal medial sides (1.1±1.3 mm and 1.5±0.9 mm, respectively; p<0.05). On postoperative radiographs, coronal tibial component angle was 88.8°±0.9° and coronal femoral component angle was 95.2°±1.6°, showing good postoperative alignment. The VAS pain score of patients significantly decreased from preoperative 3.9±0.8 to 1.1±0.9 at 6 months after the operation. One patient developed superficial tissue infection on postoperative follow-up, which was effectively treated. No other pathology was detected in the postoperative period. CONCLUSIONS Patient-specific cutting guides can provide intraoperative guidance for better placement of the implant in TKA and increase the accuracy of osteotomy and postoperative alignment.


Pulsed corona discharge oxidation of aqueous carbamazepine micropollutant.
The anti-epileptic drug carbamazepine (CBZ) receives growing attention due to slow biodegradation and inherent accumulation in the aquatic environment. The application of a gas-phase pulsed corona discharge (PCD) was investigated to remove CBZ from synthetic solutions and spiked wastewater effluent from a municipal wastewater treatment facility. The treated water was showered between high voltage (HV) wires and grounded plate electrodes, to which ultra-short HV pulses were applied. CBZ was readily oxidized and 1-(2-benzaldehyde)-4-hydroquinazoline-2-one (BQM) and 1-(2-benzaldehyde)-4-hydro-quinazoline-2,4-dione (BQD) were identified as the most abundant primary transformation products, which, contrary to CBZ ozonation data available in the literature, were further easily oxidized with PCD: BQM and BQD attributed to only a minor portion of the target compound oxidized. In concentrations commonly found in wastewater treatment plant effluents (around 5 µg L(-1)), up to 97% reduction in CBZ concentration was achieved at mere 0.3 kW h m(-3) energy consumption, and over 99.9% was removed at 1 kW h m(-3). The PCD application proved to be efficient in the removal of both the parent substance and its known transformation products, even with the competing reactions in the complex composition of wastewater.


Overt and covert dimensions of antisocial behavior in early childhood.
The present study was designed to assess both the prevalence and structure of antisocial behavior in a normative sample of preschoolers. Prevalence estimates suggested that 40% of preschoolers exhibit at least one antisocial behavior each day. Furthermore, 10% of preschoolers exhibit six or more antisocial behaviors each day. Consistent with research based on older children, factor analyses provided support for conceptualizing antisocial behavior in early childhood as consisting of both overt and covert dimensions. While both overt and covert behaviors had acceptable test-retest reliability, only overt behaviors had acceptable interrater reliability. Finally both overt and covert dimensions of antisocial behavior were uniquely related to general measures of conduct problems, hyperactivity, and adult and peer conflict in the classroom setting. Findings are discussed with regard to early assessment and the developmental course of antisocial behavior.


Adult Behavior in Male Mice Exposed to E-Cigarette Nicotine Vapors during Late Prenatal and Early Postnatal Life.
Timed-pregnant C57BL/6J mice were exposed to 2.4% nicotine in propylene glycol (PG) or 0% nicotine /PG once a day from gestational day 15 until delivery. After delivery, offspring and mothers were exposed to E-cigarette vapors for an additional 14 days from postnatal day 2 through 16. Following their last exposure serum cotinine levels were measured in female juvenile mice. Male mice underwent behavioral testing at 14 weeks of age to assess sensorimotor, affective, and cognitive functional domains. Adult male mice exposed to 2.4% nicotine/PG E-cigarette vapors had significantly more head dips in the zero maze test and higher levels of rearing activity in the open field test compared to 0% nicotine/PG exposed mice and untreated controls. In the water maze test after reversal training, the 2.4% nicotine/PG mice spent more than 25% of time in the new location whereas the other groups did not. Adult male mice exhibited increased levels of activity in the zero maze and open field tests when exposed to E-cigarette vapor containing nicotine during late prenatal and early postnatal life. These findings indicate that nicotine exposure from E-cigarettes may cause persistent behavioral changes when exposure occurs during a period of rapid brain growth.


Predictive validity in drug discovery: what it is, why it matters and how to improve it.
Successful drug discovery is like finding oases of safety and efficacy in chemical and biological deserts. Screens in disease models, and other decision tools used in drug research and development (R&D), point towards oases when they score therapeutic candidates in a way that correlates with clinical utility in humans. Otherwise, they probably lead in the wrong direction. This line of thought can be quantified by using decision theory, in which 'predictive validity' is the correlation coefficient between the output of a decision tool and clinical utility across therapeutic candidates. Analyses based on this approach reveal that the detectability of good candidates is extremely sensitive to predictive validity, because the deserts are big and oases small. Both history and decision theory suggest that predictive validity is under-managed in drug R&D, not least because it is so hard to measure before projects succeed or fail later in the process. This article explains the influence of predictive validity on R&D productivity and discusses methods to evaluate and improve it, with the aim of supporting the application of more effective decision tools and catalysing investment in their creation.


Assessment of FSHR variants and antimüllerian hormone in infertility patients with a reduced ovarian response to gonadotropin stimulation.
To study women with a poor response to ovarian hormone stimulation, known as low responders. Genetic defects in the FSH receptor gene (FSHR) were analyzed as well as antimüllerian hormone (AMH) for ovarian reserve. Retrospective cohort study. University hospital. Two hundred fifty-nine patients total: 74 low responders; 88 patients receiving assisted reproduction therapy (ART) with a normal ovarian response; and 97 women with a normal fertility status. DNA from patients was analyzed using real-time polymerase chain reaction. Serum concentrations of AMH were assessed using ELISA. The FSHR variants Asn680Ser (rs6166), Ala189Val (rs121909658), Ile160Thr (rs121909659), Thr449Ile (rs28928870) and the serum AMH concentrations were assessed. With the exception of the frequent Asn680Ser polymorphism, no homozygotic SNPs of FSHR were found. In the group of ART patients, Thr160/Ile160 variants were more frequent in comparison with women with normal fertility. The Ser680/Ser680 was more frequent in ART patients than in women with normal reproductive function. The rate of live births was markedly reduced, particularly in the low responder group. No difference was noted in the distribution of the Ala189Val and Thr449Ile variant. Low serum AMH values were observed in 75% of the low responder group. FSHR gene variations such as Asn680Ser, Ala189Val, Thr449Ile, and Ile160Thr did not seem to be a decisive factor of poor response to fertility treatment, whereas the low ovarian reserve determined by AMH is considered more crucial.


Multi-photon nanosurgery in live brain.
In the last few years two-photon microscopy has been used to perform in vivo high spatial resolution imaging of neurons, glial cells and vascular structures in the intact neocortex. Recently, in parallel to its applications in imaging, multi-photon absorption has been used as a tool for the selective disruption of neural processes and blood vessels in living animals. In this review we present some basic features of multi-photon nanosurgery and we illustrate the advantages offered by this novel methodology in neuroscience research. We show how the spatial localization of multi-photon excitation can be exploited to perform selective lesions on cortical neurons in living mice expressing fluorescent proteins. This methodology is applied to disrupt a single neuron without causing any visible collateral damage to the surrounding structures. The spatial precision of this method allows to dissect single processes as well as individual dendritic spines, preserving the structural integrity of the main neuronal arbor. The same approach can be used to breach the blood-brain barrier through a targeted photo-disruption of blood vessels walls. We show how the vascular system can be perturbed through laser ablation leading toward two different models of stroke: intravascular clot and extravasation. Following the temporal evolution of the injured system (either a neuron or a blood vessel) through time lapse in vivo imaging, the physiological response of the target structure and the rearrangement of the surrounding area can be characterized. Multi-photon nanosurgery in live brain represents a useful tool to produce different models of neurodegenerative disease.


Assessing a patient's individual risk of biopsy-detectable prostate cancer: Be aware of case mix heterogeneity and a priori likelihood.
The relation between prostate-specific antigen (PSA) and other relevant prebiopsy information is often combined in a risk calculator (RC). If the setting for RC use differs from that in which it was developed, there is a risk of making clinical decisions based on incorrect estimates of the absolute risk. The ERSPC-MRI RC predicts clinically significant prostate cancer (csPC; Gleason ≥ 3 + 4) on targeted and systematic biopsy using information on PSA, digital rectal examination, prostate volume, age, previous negative biopsy, and Prostate Imaging-Recording and Data System score. This calculator was developed on a clinical cohort of 961 men (2012-2017) with a csPC prevalence of 36%. Discrimination was good (area under the receiver operating characteristic curve 0.84). With the increasing use of multiparametric magnetic resonance imaging, we foresee that this RC will also be used for men with a lower a priori likelihood of PC. We investigated the effect of such a scenario on individual risk predictions. A small update of the intercept for the calculator can restore the accuracy to support decision-making with locally valid risk estimates. PATIENT SUMMARY: Decisions on who to refer for a prostate biopsy with its risk of sepsis and overdiagnosis require more than a prostate-specific antigen test. A prediction tool may take other relevant prebiopsy information into account, but may need to be updated to contemporary center-specific settings to provide accurate estimates of the risk of having prostate cancer.


How hyperthermophiles adapt to change their lives: DNA exchange in extreme conditions.
Transfer of DNA has been shown to be involved in genome evolution. In particular with respect to the adaptation of bacterial species to high temperatures, DNA transfer between the domains of bacteria and archaea seems to have played a major role. In addition, DNA exchange between similar species likely plays a role in repair of DNA via homologous recombination, a process that is crucial under DNA damaging conditions such as high temperatures. Several mechanisms for the transfer of DNA have been described in prokaryotes, emphasizing its general importance. However, until recently, not much was known about this process in prokaryotes growing in highly thermophilic environments. This review describes the different mechanisms of DNA transfer in hyperthermophiles, and how this may contribute to the survival and adaptation of hyperthermophilic archaea and bacteria to extreme environments.


An Investigation of Cognitive Processing of Fear Appeal Messages Promoting HPV Vaccination: Predictors and Outcomes of Magnitude and Valence of Cognitive Responses.
An improvement in HPV vaccination rates is one of the primary goals of public health organizations. Toward this end, fear appeal communication is commonly used in health interventions, warning individuals of threats of HPV infection and promoting vaccination. However, little is known about how threat-related emotions, such as fear and anxiety, influence the cognitive processing of vaccination information and how this processing is associated with vaccination intention. To address this void, this study tests a model drawing upon functional emotion theories and dual-process models of persuasion. Results from an experimental study showed that fear and anxiety, which arose from exposure to threat information, triggered motivation to process HPV protection-related information, which in turn, was positively associated with depth of HPV vaccination information processing. Subsequently, greater depth of processing led to a greater number of positive cognitive responses when participants were presented with information with a high (vs. low), level of response efficacy. Finally, greater positivity of cognitive responses predicted greater intention to obtain HPV vaccination. Collectively, our findings provide a theory-based explanation about how the sequential provision and processing of threat and efficacy information in fear appeals contribute to the promotion of HPV vaccination. Implications for designing fear appeal messages are discussed.


Histochemical analyses of tissue-engineered human menisci.
The field of tissue engineering remains one of the least explored areas of current meniscal research but holds great promise. In this investigation, meniscal fibrochondrocytes were isolated from fresh human meniscal tissue and seeded onto synthetic polyglycolic acid (PGA) scaffolds. Constructs were implanted into the dorsal subcutaneous space of athymic nude mice. Control scaffolds, devoid of meniscal cells, were simultaneously implanted in additional mice. Constructs were harvested over 12 weeks and treated with a variety of histochemical stains to analyze general specimen morphology, cellular viability and proliferation, and collagen secretion. Results indicate that meniscal fibrochondrocyte proliferation increased over the time of implantation with cellular consolidation occurring as the PGA scaffolding was progressively hydrolyzed. Collagen production also increased over time. There were favorable similarities between constructs and human meniscal controls in terms of cellular morphology, phenotypic expression, and collagen production. These initial findings demonstrate procedures supporting proliferation of meniscal fibrochondrocytes, expression of fibrochondral phenotype, and the formation of putative meniscal tissue.


Low life jacket use among adult recreational boaters: a qualitative study of risk perception and behavior factors.
Life jackets may prevent one in two drowning deaths, however, 85% of recreational boating-related drowning victims in the United States in 2012 did not wear a life jacket. This study explored behavioral factors and strategies to encourage consistent life jacket use among adult recreational boaters. We conducted a qualitative study among boat owners who boat regularly, and explored factors associated with life jacket use by adults and child or adolescent passengers. Sixteen boaters participated in four focus groups. Most boaters reported inconsistent use of life jackets, using them only when conditions were poor. Each described episodes of unpredictable boating risk which occurred despite favorable conditions. Most required younger child passengers to wear a life jacket, but reported resistance among older children. Barriers to consistent life jacket use included discomfort and the belief that life jacket use indicated inexperience or poor swimming ability. Participants stated that laws requiring life jacket use would change behavior especially for children. The only demonstrated behavior change among group members was associated with use of inflatable life jacket devices. Boating risk is inherently unpredictable; therefore interventions should focus on strategies for increasing consistent use of life jackets. Passage and enforcement of life jacket legislation for older children and adults is likely a promising approach for behavior change. Designing more comfortable, better-fitting, more appealing life jackets will be paramount to encouraging consistent use.


Farm income and production impacts of using GM crop technology 1996-2016.
This paper estimates the value of using genetically modified (GM) crop technology in agriculture at the farm level. It follows and updates earlier annual studies which examined impacts on yields, key variable costs of production, direct farm (gross) income and impacts on the production base of the four main crops of soybeans, corn, cotton and canola. The commercialisation of GM crops has occurred at a rapid rate since the mid 1990s, with important changes in both the overall level of adoption and impact occurring in 2016. This annual updated analysis shows that there continues to be very significant net economic benefits at the farm level amounting to $18.2 billion in 2016 and $186.1 billion for the period 1996-2016 (in nominal terms). These gains have been divided 48% to farmers in developed countries and 52% to farmers in developing countries. About 65% of the gains have derived from yield and production gains with the remaining 35% coming from cost savings. The technology has also made important contributions to increasing global production levels of the four main crops, having, for example, added 213 million tonnes and 405 million tonnes respectively, to the global production of soybeans and maize since the introduction of the technology in the mid 1990s.


Young domestic chicks spontaneously represent the absence of objects.
Absence is a notion that is usually captured by language-related concepts like zero or negation. Whether nonlinguistic creatures encode similar thoughts is an open question, as everyday behavior marked by absence (of food, of social partners) can be explained solely by expecting presence somewhere else. We investigated 8-day-old chicks' looking behavior in response to events violating expectations about the presence or absence of an object. We found different behavioral responses to violations of presence and absence, suggesting distinct underlying mechanisms. Importantly, chicks displayed an avian signature of novelty detection to violations of absence, namely a sex-dependent left-eye-bias. Follow-up experiments excluded accounts that would explain this bias by perceptual mismatch or by representing the object at different locations. These results suggest that the ability to spontaneously form representations about the absence of objects likely belongs to the initial cognitive repertoire of vertebrate species.


CD9-positive microvesicles mediate the transfer of molecules to Bovine Spermatozoa during epididymal maturation.
Acquisition of fertilization ability by spermatozoa during epididymal transit occurs in part by the transfer of molecules from membranous vesicles called epididymosomes. Epididymosomes are heterogeneous in terms of both size and molecular composition. Exosomes and other related small membranous vesicles (30-120 nm) containing tetraspanin proteins on their surface are found in many biological fluids. In this study, we demonstrate that these vesicles are present in bovine cauda epididymal fluid as a subpopulation of epididymosomes. They contain tetraspanin CD9 in addition to other proteins involved in sperm maturation such as P25b, GliPr1L1, and MIF. In order to study the mechanism of protein transfer to sperm, DilC12-labeled unfractionated epididymosomes or CD9-positive microvesicles were coincubated with epididymal spermatozoa, and their transfer was evaluated by flow cytometry. CD9-positive microvesicles from epididymal fluid specifically transferred molecules to spermatozoa, whereas those prepared from blood were unable to do so. The CD9-positive microvesicles transferred molecules to the same sperm regions (acrosome and midpiece) as epididymosomes, with the same kinetics; however, the molecules were preferentially transferred to live sperm and, in contrast to epididymosomes, Zn(2+) did not demonstrate potentiated transfer. Tetraspanin CD9 was associated with other proteins on the membrane surface of CD9-positive microvesicles according to coimmunoprecipitation experiments. CD26 cooperated with CD9 in the molecular transfer to sperm since the amount of molecules transferred was significantly reduced in the presence of specific antibodies. In conclusion, CD9-positive microvesicles are present in bovine cauda epididymal fluid and transfer molecules to live maturing sperm in a tissue-specific manner that involves CD9 and CD26.