Percutaneous Mesh Expansion: A Regenerative Wound Closure Alternative.
Puncture wounds in the 1-mm range usually heal without scars. Stacking rows of these punctures offers a scarless method to generate tissue by mesh expansion. The authors developed a percutaneous mesh expansion procedure and present their experience for its wound closure application. Over a 6-year period, the authors applied percutaneous mesh expansion to 65 consecutive patients aged 58 to 101 years (mean, 72 years) with 67 full-thickness calvarial defects ranging in size from 2.5 × 3 cm to 7 × 8 cm (mean, 14 cm) that would have all required flaps for closure. Thirty-six were still anticoagulated, and 20 had prior scalp resections. After tumescent epinephrine anesthesia, the authors temporarily approximate the wound by placing it under strong tension. Using 1.1-mm cutting point needles that selectively sever tissues under tension, the authors inflict rows of staggered alternating punctures over a distance five times the defect width. This results in 20 percent expansion of the meshed area, generating the tissue necessary for defect coverage. When the tension is completely released, closure is performed with simple sutures or staples. The authors avoid overmeshing, especially close to the wound edges, and perform no undermining or additional incisions. At 6-week follow-up, all defects were healed with only a straight resection scar. However, of the 10 defects larger than 5 × 5 cm, five had wound healing delay and three required a small skin graft. No other complication was observed. Percutaneous mesh expansion is a minimally invasive procedure that harnesses the body's natural capabilities to regenerate across small gaps. It sums these regenerated gaps in a mesh pattern that expands tissues to close complex wounds without flaps or additional incisions. Therapeutic, IV.


The Perceived Stigma of Mental Health Services Among Rural Parents of Children With Psychosocial Concerns.
To examine parents' perceptions of stigma regarding mental health services for their child, consider stigma in the context of novel service delivery settings (e.g., telehealth, primary care, and schools), and evaluate stigma with other factors known to influence service access. 347 caregivers of children with psychosocial concerns completed surveys regarding their perceptions of stigma, service delivery settings, and barriers to care. Parents endorsed low levels of stigma around services. Greater perceived stigma was related to less willingness to seek services in a mental/behavioral health center or schools but not in other settings, even when other barriers were considered. Having a younger child and a history of prior services was associated with greater willingness to seek services. Stigma does appear to present as a barrier, but only for some parents. Providing mental health services to young children and their parents in some nontraditional settings may increase access.


Stalk cell formation in monolayers from isolated prestalk and prespore cells of Dictyostelium discoideum: evidence for two populations of prestalk cells.
Cells from the pseudoplasmodial stage of Dictyostelium discoideum differentiation were dispersed and separated on Percoll gradients into prestalk and prespore cells. The requirements for stalk cell formation in low-density monolayers from the two cell types were determined. The isolated prespore cells required both the Differentiation Inducing Factor (DIF) and cyclic AMP for stalk cell formation. In contrast, only part of the isolated prestalk cell population required both cyclic AMP and DIF, the remainder requiring DIF alone, suggesting the possibility that there were two populations of prestalk cells, one independent of cyclic AMP and one dependent on cyclic AMP for stalk cell formation. The finding that part of the prestalk cell population required only a brief incubation in the presence of DIF to induce stalk cell formation, whilst the remainder required a considerably longer incubation in the presence of both DIF and cyclic AMP was consistent with this idea. In addition, stalk cell formation from cyclic-AMP-dependent prestalk cells was relatively more sensitive to caffeine inhibition than stalk cell formation from cyclic-AMP-independent prestalk cells. The latter cells were enriched in the most anterior portion of the migrating pseudoplasmodium, indicating that there is spatial segregation of the two prestalk cell populations. The conversion of prespore cells to stalk cells took longer and was more sensitive to caffeine when compared to stalk cell formation from cyclic-AMP-dependent prestalk cells.


Adolescents exhibit reduced Pavlovian biases on instrumental learning.
Multiple learning systems allow individuals to flexibly respond to opportunities and challenges present in the environment. An evolutionarily conserved "Pavlovian" learning mechanism couples valence and action, promoting a tendency to approach cues associated with reward and to inhibit action in the face of anticipated punishment. Although this default response system may be adaptive, these hard-wired reactions can hinder the ability to learn flexible "instrumental" actions in pursuit of a goal. Such constraints on behavioral flexibility have been studied extensively in adults. However, the extent to which these valence-specific response tendencies bias instrumental learning across development remains poorly characterized. Here, we show that while Pavlovian response biases constrain flexible action learning in children and adults, these biases are attenuated in adolescents. This adolescent-specific reduction in Pavlovian bias may promote unbiased exploration of approach and avoidance responses, facilitating the discovery of rewarding behavior in the many novel contexts that adolescents encounter.


Loss of MCL-1 leads to impaired autophagy and rapid development of heart failure.
Myeloid cell leukemia-1 (MCL-1) is an anti-apoptotic BCL-2 protein that is up-regulated in several human cancers. MCL-1 is also highly expressed in myocardium, but its function in myocytes has not been investigated. We generated inducible, cardiomyocyte-specific Mcl-1 knockout mice and found that ablation of Mcl-1 in the adult heart led to rapid cardiomyopathy and death. Although MCL-1 is known to inhibit apoptosis, this process was not activated in MCL-1-deficient hearts. Ultrastructural analysis revealed disorganized sarcomeres and swollen mitochondria in myocytes. Mitochondria isolated from MCL-1-deficient hearts exhibited reduced respiration and limited Ca(2+)-mediated swelling, consistent with opening of the mitochondrial permeability transition pore (mPTP). Double-knockout mice lacking MCL-1 and cyclophilin D, an essential regulator of the mPTP, exhibited delayed progression to heart failure and extended survival. Autophagy is normally induced by myocardial stress, but induction of autophagy was impaired in MCL-1-deficient hearts. These data demonstrate that MCL-1 is essential for mitochondrial homeostasis and induction of autophagy in the heart. This study also raises concerns about potential cardiotoxicity for chemotherapeutics that target MCL-1.


A multigene phylogeny of the fly superfamily Asiloidea (Insecta): Taxon sampling and additional genes reveal the sister-group to all higher flies (Cyclorrhapha).
Asiloidea are a group of 9 lower brachyceran fly families, considered to be the closest relative to the large Metazoan radiation Eremoneura (Cyclorrhapha+Empidoidea). The evidence for asiloid monophyly is limited, and few characters define the relationships between the families of Asiloidea and Eremoneura. Additionally, enigmatic genera, Hilarimorpha and Apystomyia, retain morphological characters of both asiloids and higher flies. We use the nuclear protein-coding gene CAD and 28S rDNA to test the monophyly of Asiloidea and to resolve its relationship to Eremoneura. We explore the effects of taxon sampling on support values and topological stability, the resolving power of additional genes, and hypothesis testing using four-cluster likelihood mapping. We find that: (1) the 'asiloid' genus Apystomyia is sister to Cyclorrhapha, (2) the remaining asiloids are monophyletic at the exclusion of the family Bombyliidae, and (3) our best estimate of relationships places the asiloid flies excluding Bombyliidae as the sister-group to Eremoneura, though high support is lacking.


Pre-stimulus Brain Activity Is Associated With State-Anxiety Changes During Single-Session Transcranial Direct Current Stimulation.
Transcranial direct current stimulation is a promising neuromodulation method for treating depression. However, compared with pharmacological treatment, previous studies have reported that a relatively limited proportion of patients respond to tDCS treatment. In addition, the neurophysiological mechanisms underlying tDCS treatment remain unclear, making it difficult to identify response predictors for tDCS treatment based on neurophysiological function. Because treatment effects are achieved by repetitive application of tDCS, studying the immediate effects of tDCS in depressive patients could extend understanding of its treatment mechanisms. However, immediate changes in a single session of tDCS are not well documented. Thus, in the current study, we focused on the immediate impact of tDCS and its association with pre-stimulus brain activity. To address this question, we applied anodal tDCS to the left dorsolateral prefrontal cortex (DLPFC) or dorsomedial prefrontal cortex (DMPFC) in 14 patients with major depressive disorder (MDD) and 19 healthy controls (HCs), at an intensity of 1.0 mA for 20 min in a single session. To evaluate anxiety, the state trait anxiety inventory was completed before and after tDCS. We recorded resting electroencephalography before tDCS, and calculated electrical neuronal activity in the theta and alpha frequency bands using standardized low-resolution electromagnetic tomography. We found that, during application of left DLPFC tDCS to patients with MDD, the anxiety reduction effect of tDCS was related to higher baseline theta-band activity in the rostral anterior cingulate cortex (rACC) and no medication with benzodiazepine used as hypnotic. For DMPFC stimulation in MDD, the anxiety reduction effect was associated with lower baseline alpha-band activity in the left inferior parietal lobule. In contrast, in HCs, the anxiety reduction effect was associated with higher baseline alpha activity in the precuneus during DMPFC stimulation. The current results suggest that the association between pre-tDCS brain activity and the anxiety reduction effect of tDCS depends on psychopathology (depressed or non-depressed) as well as the site of stimulation (DMPFC or left DLPFC) and insomnia. Furthermore, the results suggest that tDCS response might be associated with baseline resting state electrophysiological neural activity.


Test positivity for Maedi-Visna virus and Mycobacterium avium ssp. paratuberculosis in Sarda ewes: Effects on milk composition and coagulation traits and heritability estimates for susceptibility.
Maedi-Visna virus (MVV) and Mycobacterium avium ssp. paratuberculosis (MAP) are two pathogens that cause chronic, production-limiting diseases in dairy sheep. Although they are present worldwide, there are no detailed reports on their actual effects on milk traits in the literature. This study was designed to investigate the effects of test positivity to MVV and MAP on ovine milk yield, composition and coagulation properties, and curd-firming over time (CFt) variables in clinically healthy animals at the field level. The additive genetic variation and heritabilities of MVV and MAP positivity were also estimated. Milk samples were collected from 1,079 Sarda sheep kept on 23 farms, and pedigree information was obtained from the flock book. Milk yield was also recorded on the sampling date. Positivity for MVV and MAP was determined from milk samples using indirect ELISA test kits. Milk composition traits were measured by spectroscopy, milk coagulation properties were measured with a Formagraph (Foss Italia, Padua, Italy), and CFt traits were calculated using the data from the Formagraph diagram. The effects of MVV and MAP positivity on milk traits were determined through a set of mixed linear models, which took into account various sources of variation, such as days in milk, parity, and flock effects, and included the effects (positive or negative) of the 2 pathogens. A Bayesian threshold sire model with sire relationship was used to estimate genetic variation and heritability. The overall animal prevalence of MVV-positive ewes was 43.6%; on only 1 farm of the 23 tested were all sampled ewes negative. An overall animal prevalence of 10.6% was recorded for MAP, with 4 farms at 0%. Positivity for MVV significantly affected the logarithmic score of the bacterial count, curd firmness after 30 min and 45 min, and the curd-firming instant rate constant. We found significant effects of MAP infection on milk composition, pH, and rennet coagulation time. The mean of the posterior distributions of heritability estimates on the liability scale was 0.15 for MAP and 0.07 for MVV. Our results demonstrate that only a few traits are negatively affected by MVV and MAP positivity, and that there is exploitable genetic variation in MVV and MAP susceptibility in dairy sheep.


Moral character predominates in person perception and evaluation.
What sorts of trait information do people most care about when forming impressions of others? Recent research in social cognition suggests that "warmth," broadly construed, should be of prime importance in impression formation. Yet, some prior research suggests that information about others' specifically moral traits--their moral "character"--may be a primary dimension. Although warmth and character have sometimes been conceived of as interchangeable, we argue that they are separable, and that across a wide variety of contexts, character is usually more important than warmth in impression formation. We first showed that moral character and social warmth traits are indeed separable (Studies 1 and 2). Further studies that used correlational and experimental methods showed that, as predicted, in most contexts, moral character information is more important in impression formation than is warmth information (Studies 2-6). Character information was also more important than warmth information with respect to judgments of traits' perceived fundamentalness to identity, their uniquely human quality, their context-independence, and their controllability (Study 2). Finally, Study 7 used an archival method to show that moral character information appears more prominently than warmth information in obituaries, and more strongly determines the impressions people form of the individuals described in those obituaries. We discuss implications for current theories of person perception and social cognition.


A nutrient-sensitive restriction point is active during retinal progenitor cell differentiation.
In many growing tissues, slowly dividing stem cells give rise to rapidly proliferating progenitors that eventually exit the cell cycle and differentiate. Growth rates are limited by nutrient availability, but it is unclear which steps of the proliferation-differentiation programme are particularly sensitive to fuel supplies. We examined how nutrient deprivation (ND) affects stem and progenitor cells in the ciliary marginal zone (CMZ) of the amphibian retina, a well-characterised neurogenic niche. We show that ND specifically blocks the proliferation and differentiation of progenitor cells through an mTOR-mediated mechanism. By contrast, the identity and proliferation of retinal stem cells are insensitive to ND and mTOR inhibition. Re-feeding starved retinas in vitro rescues both proliferation and differentiation, and activation of mTOR is sufficient to stimulate differentiation even in ND retinas. These results suggest that an mTOR-mediated restriction point operates in vivo to couple nutrient abundance to the proliferation and differentiation programme in retinal progenitor cells.


Spatiotemporal temperature distribution and cancer cell death in response to extracellular hyperthermia induced by gold nanorods.
Plasmonic nanoparticles have shown promise in hyperthermic cancer therapy, both in vitro and in vivo. Previous reports have described hyperthermic ablation using targeted and nontargeted nanoparticles internalized by cancer cells, but most reports do not describe a theoretical analysis for determining optimal parameters. The focus of the current research was first to evaluate the spatiotemporal temperature distribution and cell death induced by extracellular hyperthermia in which gold nanorods (GNRs) were maintained in the dispersion outside human prostate cancer cells. The nanorod dispersion was irradiated with near-infrared (NIR) laser, and the spatiotemporal distribution of temperature was determined experimentally. This information was employed to develop and validate theoretical models of spatiotemporal temperature profiles for gold nanorod dispersions undergoing laser irradiation and the impact of the resulting heat generation on the viability of human prostate cancer cells. A cell injury/death model was then coupled to the heat transfer model to predict spatial and temporal variations in cell death and injury. The model predictions agreed well with experimental measurements of both temperature and cell death profiles. Finally, the model was extended to examine the impact of selective binding of gold nanorods to cancer cells compared to nonmalignant cells, coupled with a small change in cell injury activation energy. The impact of these relatively minor changes results in a dramatic change in the overall cell death rate. Taken together, extracellular hyperthermia using gold nanorods is a promising strategy, and tailoring the cellular binding efficacy of nanorods can result in varying therapeutic efficacies using this approach.


Novel approach of MALDI drug imaging, immunohistochemistry, and digital image analysis for drug distribution studies in tissues.
Drug efficacy strongly depends on the presence of the drug substance at the target site. As vascularization is an important factor for the distribution of drugs in tissues, we analyzed drug distribution as a function of blood vessel localization in tumor tissue. To explore distribution of the anticancer drugs afatinib, erlotinib, and sorafenib, a combined approach of matrix-assisted laser desorption/ionization (MALDI) drug imaging and immunohistochemical vessel staining was applied and examined by digital image analysis. The following two xenograft models were investigated: (1) mice carrying squamous cell carcinoma (FaDu) xenografts (ntumor = 13) were treated with afatinib or erlotinib, and (2) sarcoma (A673) xenograft bearing mice (ntumor = 8) received sorafenib treatment. MALDI drug imaging revealed a heterogeneous distribution of all anticancer drugs. The tumor regions containing high drug levels were associated with a higher degree of vascularization than the regions without drug signals (p < 0.05). When correlating the impact of blood vessel size to drug abundance in the sarcoma model, a higher amount of small vessels was detected in the tumor regions with high drug levels compared to the tumor regions with low drug levels (p < 0.05). With the analysis of coregistered MALDI imaging and CD31 immunohistochemical data by digital image analysis, we demonstrate for the first time the potential of correlating MALDI drug imaging and immunohistochemistry. Here we describe a specific and precise approach for correlating histological features and pharmacokinetic properties of drugs at microscopic level, which will provide information for the improvement of drug design, administration formula or treatment schemes.


[Changing epidemiology of infections in the Netherlands in 2008/'09].
To describe the epidemiological characteristics of Clostridium difficile infection (CDI) in the Netherlands. Descriptive. After the first outbreaks in 2005 of Clostridium difficile infection (CDI) due to the hypervirulent PCR ribotype 027, a national reference laboratory was started in the Leiden University Medical Centre for typing and characterisation of C. difficile. Data were obtained from this national reference laboratory and from a continuous surveillance in 14 Dutch hospitals. The study period was January 2008-June 2009. In 2008, the incidence of CDI was 18 per 10.000 admissions (range: 14-23) in the 14 participating hospitals in the national surveillance study. In the study period, a total of 1867 cases of CDI were reported from 63 centres. The number of CDI outbreaks caused by type 027 decreased in the period January 2008 to June 2009, compared to the preceding years: type 027 was the cause of outbreaks in 4 Dutch hospitals in 2008-2009, whereas all 14 hospitals experienced an outbreak due to this type in the period 2005-2007. Although no systemic surveillance has been carried out in nursing homes, 24 Dutch nursing homes reported outbreaks of C. difficile in the period 2005-2009, in 12 of which type 027 was isolated. There was an increase of CDI detected in patients with diarrhoea outside health care facilities which appeared to coincide with the emergence of other types. In particular, type 078 increased since the end of 2006 and became the third most frequent type in the Netherlands. This type has also been recognised since 2007 as an important cause of CDI infection in animals, especially pigs. Recently there have also been reports that at least a third of the patients with CDI outside health care institutions do not have known risk factors for CDI such as an underlying disease, recent hospitalization or antibiotic use. CDI can also develop in the community The general practitioner should consider C. difficile in the differential diagnosis of elderly patients with diarrhoea.


A fuzzy expert system for diabetes decision support application.
An increasing number of decision support systems based on domain knowledge are adopted to diagnose medical conditions such as diabetes and heart disease. It is widely pointed that the classical ontologies cannot sufficiently handle imprecise and vague knowledge for some real world applications, but fuzzy ontology can effectively resolve data and knowledge problems with uncertainty. This paper presents a novel fuzzy expert system for diabetes decision support application. A five-layer fuzzy ontology, including a fuzzy knowledge layer, fuzzy group relation layer, fuzzy group domain layer, fuzzy personal relation layer, and fuzzy personal domain layer, is developed in the fuzzy expert system to describe knowledge with uncertainty. By applying the novel fuzzy ontology to the diabetes domain, the structure of the fuzzy diabetes ontology (FDO) is defined to model the diabetes knowledge. Additionally, a semantic decision support agent (SDSA), including a knowledge construction mechanism, fuzzy ontology generating mechanism, and semantic fuzzy decision making mechanism, is also developed. The knowledge construction mechanism constructs the fuzzy concepts and relations based on the structure of the FDO. The instances of the FDO are generated by the fuzzy ontology generating mechanism. Finally, based on the FDO and the fuzzy ontology, the semantic fuzzy decision making mechanism simulates the semantic description of medical staff for diabetes-related application. Importantly, the proposed fuzzy expert system can work effectively for diabetes decision support application.


H2G-Net: A multi-resolution refinement approach for segmentation of breast cancer region in gigapixel histopathological images.
Over the past decades, histopathological cancer diagnostics has become more complex, and the increasing number of biopsies is a challenge for most pathology laboratories. Thus, development of automatic methods for evaluation of histopathological cancer sections would be of value. In this study, we used 624 whole slide images (WSIs) of breast cancer from a Norwegian cohort. We propose a cascaded convolutional neural network design, called H2G-Net, for segmentation of breast cancer region from gigapixel histopathological images. The design involves a detection stage using a patch-wise method, and a refinement stage using a convolutional autoencoder. To validate the design, we conducted an ablation study to assess the impact of selected components in the pipeline on tumor segmentation. Guiding segmentation, using hierarchical sampling and deep heatmap refinement, proved to be beneficial when segmenting the histopathological images. We found a significant improvement when using a refinement network for post-processing the generated tumor segmentation heatmaps. The overall best design achieved a Dice similarity coefficient of 0.933±0.069 on an independent test set of 90 WSIs. The design outperformed single-resolution approaches, such as cluster-guided, patch-wise high-resolution classification using MobileNetV2 (0.872±0.092) and a low-resolution U-Net (0.874±0.128). In addition, the design performed consistently on WSIs across all histological grades and segmentation on a representative × 400 WSI took ~ 58 s, using only the central processing unit. The findings demonstrate the potential of utilizing a refinement network to improve patch-wise predictions. The solution is efficient and does not require overlapping patch inference or ensembling. Furthermore, we showed that deep neural networks can be trained using a random sampling scheme that balances on multiple different labels simultaneously, without the need of storing patches on disk. Future work should involve more efficient patch generation and sampling, as well as improved clustering.


The Krüppel-like factor 9 cistrome in mouse hippocampal neurons reveals predominant transcriptional repression via proximal promoter binding.
Krüppel-like factor 9 (Klf9) is a zinc finger transcription factor that functions in neural cell differentiation, but little is known about its genomic targets or mechanism of action in neurons. We used the mouse hippocampus-derived neuronal cell line HT22 to identify genes regulated by Klf9, and we validated our findings in mouse hippocampus. We engineered HT22 cells to express a Klf9 transgene under control of the tetracycline repressor, and used RNA sequencing to identify genes modulated by Klf9. We found 217 genes repressed and 21 induced by Klf9. We also engineered HT22 cells to co-express biotin ligase and a Klf9 fusion protein containing an N-terminal biotin ligase recognition peptide. Using chromatin-streptavidin precipitation (ChSP) sequencing we identified 3,514 genomic regions where Klf9 associated. Seventy-five percent of these were within 1 kb of transcription start sites, and Klf9 associated in chromatin with 60% of the repressed genes. We analyzed the promoters of several repressed genes containing Klf9 ChSP peaks using transient transfection reporter assays and found that Klf9 repressed promoter activity, which was abolished after mutation of Sp/Klf-like motifs. Knockdown or knockout of Klf9 in HT22 cells caused dysregulation of Klf9 target genes. Chromatin immunoprecipitation assays showed that Klf9 associated in chromatin from mouse hippocampus with genes identified by ChSP sequencing on HT22 cells, and expression of Klf9 target genes was dysregulated in the hippocampus of neonatal Klf9-null mice. Gene ontology analysis revealed that Klf9 genomic targets include genes involved in cystokeletal remodeling, Wnt signaling and inflammation. We have identified genomic targets of Klf9 in hippocampal neurons and created a foundation for future studies on how it functions in chromatin, and regulates neuronal morphology and survival across the lifespan.


Hyperimmune bovine colostrum as a low-cost, large-scale source of antibodies with broad neutralizing activity for HIV-1 envelope with potential use in microbicides.
Bovine colostrum (first milk) contains very high concentrations of IgG, and on average 1 kg (500 g/liter) of IgG can be harvested from each immunized cow immediately after calving. We used a modified vaccination strategy together with established production systems from the dairy food industry for the large-scale manufacture of broadly neutralizing HIV-1 IgG. This approach provides a low-cost mucosal HIV preventive agent potentially suitable for a topical microbicide. Four cows were vaccinated pre- and/or postconception with recombinant HIV-1 gp140 envelope (Env) oligomers of clade B or A, B, and C. Colostrum and purified colostrum IgG were assessed for cross-clade binding and neutralization against a panel of 27 Env-pseudotyped reporter viruses. Vaccination elicited high anti-gp140 IgG titers in serum and colostrum with reciprocal endpoint titers of up to 1 × 10(5). While nonimmune colostrum showed some intrinsic neutralizing activity, colostrum from 2 cows receiving a longer-duration vaccination regimen demonstrated broad HIV-1-neutralizing activity. Colostrum-purified polyclonal IgG retained gp140 reactivity and neutralization activity and blocked the binding of the b12 monoclonal antibody to gp140, showing specificity for the CD4 binding site. Colostrum-derived anti-HIV antibodies offer a cost-effective option for preparing the substantial quantities of broadly neutralizing antibodies that would be needed in a low-cost topical combination HIV-1 microbicide.


Frequency of citrus fruit intake is associated with the incidence of cardiovascular disease: the Jichi Medical School cohort study.
It has been reported that fruit intake protects against cardiovascular disease (CVD). However, most of the relevant studies were conducted in Western countries, and only a few investigated Japanese populations. The present cohort study assessed the effect of citrus fruit intake on the incidence of CVD and its subtypes in a Japanese population. A baseline examination consisting of physical and blood examinations and a self-administered questionnaire was conducted during the period from April 1992 through July 1995. Dietary habits were assessed using a food frequency questionnaire that was divided into 5 categories. Citrus fruit was examined separately due to its frequent consumption by the general Japanese population. Using the Cox proportional hazards model, data from 10,623 participants (4147 men, 6476 women) who had no history of CVD or carcinoma were analyzed to assess the association between frequency of citrus fruit intake and CVD incidence. Frequent intake of citrus fruit was associated with a lower incidence of CVD: the hazard ratio for almost daily intake versus infrequent intake of citrus fruit was 0.57 (95% confidence interval: 0.33-1.01, P for trend = 0.04) in men and 0.51 (0.29-0.88, P for trend = 0.02) in women. Frequent intake of citrus fruit was also associated with lower incidences of both all stroke and cerebral infarction, but not hemorrhagic stroke or myocardial infarction. Frequent intake of citrus fruit may reduce the incidence of CVD, especially cerebral infarction, in men and women.


Evaluation of a hybrid artificial liver module based on a spheroid culture system of embryonic stem cell-derived hepatic cells.
Hybrid artificial liver (HAL) is an extracorporeal circulation system comprised of a bioreactor containing immobilized functional liver cells. It is expected to not only serve as a temporary liver function support system, but also to accelerate liver regeneration in recovery from hepatic failure. One of the most difficult problems in developing a hybrid artificial liver is obtaining an adequate cell source. In this study, we attempt to differentiate embryonic stem (ES) cells by hepatic lineage using a polyurethane foam (PUF)/spheroid culture in which the cultured cells spontaneously form spherical multicellular aggregates (spheroids) in the pores of the PUF. We also demonstrate the feasibility of the PUF-HAL system by comparing ES cells to primary hepatocytes in in vitro and ex vivo experiments. Mouse ES cells formed multicellular spheroids in the pores of PUF. ES cells expressed liver-specific functions (ammonia removal and albumin secretion) after treatment with the differentiation-promoting agent, sodium butyrate (SB). We designed a PUF-HAL module comprised of a cylindrical PUF block with many medium-flow capillaries for hepatic differentiation of ES cells. The PUF-HAL module cells expressed ammonia removal and albumin secretion functions after 2 weeks of SB culture. Because of high proliferative activity of ES cells and high cell density, the maximum expression level of albumin secretion function per unit volume of module was comparable to that seen in primary mouse hepatocyte culture. In the animal experiments with rats, the PUF-HAL differentiating ES cells appeared to partially contribute to recovery from liver failure. This outcome indicates that the PUF module containing differentiating ES cells may be a useful biocomponent of a hybrid artificial liver support system.


Online psychoeducation and digital assessments as a first step of treatment for borderline personality disorder: A protocol for a pilot randomized controlled trial.
Treatment trials for borderline personality disorder (BPD) have consistently demonstrated that approaches that are diagnostically tailored are superior to those which are not. Currently, gold standard treatments for BPD are highly intensive, lengthy, and specialized, leading to a critical gap between the supply and demand of effective, evidence-based treatment for patients who receive a diagnosis of BPD. Psychoeducation, which is a common component of most treatments known to be effective, is a low-cost, low-burden intervention proven to relieve symptoms. The present study builds on psychoeducation research, assessing online video prescriptions as a means of disseminating information patients need to know about their diagnosis and care. This article presents the study protocol for a safety, feasibility, and preliminary efficacy trial of psychoeducational video prescriptions and online assessment with feedback for newly diagnosed individuals with BPD. We aim to recruit 100 adults recently diagnosed with BPD to be randomly assigned to receive videos about BPD or videos about non-BPD mental health topics that are matched in length in the first step of the study. All participants will complete daily surveys about their emotions, interpersonal interactions, and behaviors, as well as self-report assessments and cognitive tests at 4 different time points. Half of the participants in the intervention group will receive feedback on their symptom ratings and cognitive test performance to assess whether there is incremental value in tailoring this online set of interventions with individualized feedback unique to each participant. This study aims to assess the effects of BPD-focused psychoeducational videos with and without personalized feedback, on BPD and depressive symptom severity as well as core mechanisms of the disorder such as loneliness, rejection sensitivity, cognitive control difficulties, and self-clarity. Results will inform efforts to progress to a larger, more definitive trial. Clinical trials registration: The protocol is registered with ClinicalTrials.gov NCT05358925.


Investigating digit ratio (2D:4D) in a highly male-dominated occupation: the case of firefighters.
Second-to-fourth digit ratio (2D:4D), a widely studied putative marker for masculinization through prenatal androgen exposure, is lower (more masculinized) in athletes than in general population controls, and athletes with lower 2D:4D have higher sporting success. Occupations differ markedly in perceived masculinity and actual maleness (sex ratios), but these givens have not yet been picked up and utilized in 2D:4D research. Accordingly, this study extended existing accounts on 2D:4D in athletes to a novel approach: 2D:4D and possible relationships to a variety of candidate variables (demographic, fertility-related, psychological, and other) were investigated in firefighters, a highly male-dominated occupation. Contrary to expectation, 2D:4D in firefighters (N = 134) was not lower than in local male population controls. Lower 2D:4D corresponded to lower service ranks. Replicating previous findings either unequivocally or partly, lower 2D:4D was associated with larger family size, later sibling position, left-handedness, and higher scores in the disinhibition component of sensation seeking. Not replicating prior evidence, 2D:4D was unrelated to body-mass index, offspring sex ratio, and sporting performance level. Novel findings included low 2D:4D in those with low relationship satisfaction and in cigarette smokers, especially among heavy smokers. Absolute finger length, a positive correlate of pubertal-adolescent androgen levels, was also considered. This marker showed negative associations with relationship consensus and satisfaction and positive ones with perceived quality of relationship alternatives and the experience seeking component of sensation seeking. The merits of this additional marker, relative to 2D:4D, for supplementing studies of possible sex-hormonal effects on personality and directions for future inquiry along these lines are discussed.


[Association between mannose-binding lectin 2 gene and protein kinase C-beta 1 gene polymorphisms and type 2 diabetic macrovascular complications in northern Chinese Han population].
To assess the association between single nucleotide polymorphisms (SNPs) of mannose-binding lectin 2 gene (MBL2) (rs1800450, rs1800451 and rs11003125) and protein kinase C-beta 1 gene (PRKC beta 1) (rs3700106, rs2575390) with diabetic macroangiopathy in northern Chinese Han population. The samples have included 318 type 2 diabetes mellitus (T2DM) patients and 448 normoglycemic controls. The five SNPs were determined by a Multiplex SnaPshot method. Biochemical indices such as fasting plasma-glucose, triglyceride and total cholesterol were also measured. Linkage disequilibrium and haplotype analysis were carried out for all samples using Haploview 4.2. Additive model was applied to assess the effect of interaction between SNPs and environment factors on macrovascular complications. Genotypic frequencies of rs11003125 have differed significantly between the controls and patients with coronary heart disease and peripheral vascular disease (P=0.024 and 0.004, respectively). The allele frequency of rs11003125 was also statistically significant between the two groups (P=0.014 and 0.001, respectively). Compared with patients without macrovascular complications, the allele frequency of rs11003125 was significantly different in patients with peripheral vascular disease (P=0.031). No significant differences were found between the distribution of the genotype frequency and allele frequencies of other variants. Haplotype analysis indicated that, compared with controls and patients without macrovascular complications, individuals with G allele of rs1800450 and C allele of rs11003125 had a higher risk for macrovascular complications. The rs11003125 polymorphism located in the promoter region of MBL2 gene is associated with macrovascular complications of T2DM in northern Chinese Han population. G allele of rs1800450 and C allele of rs11003125 may be risk factors for macrovascular complications. There were additive interactive effects for rs11003125 polymorphism (GC+CC) and hypertension, diabetic nephropathy, diabetic neuropathy and diabetic retinopathy on macrovascular complications.


Preliminary studies of magnetic field facilitation of electric conduction in electrically switched "on" dye films that may be room-temperature superconductors.
Some organic dyes of a double diazo structure, pressurized into thin films, show trapping of magnetic flux at room temperature, suggestive of the behavior of type 2 superconductors. Some also show switching from low to high electrical conductivity at a voltage threshold, like amorphous inorganic semiconductors. An applied magnetic field facilitates switching, as well as current flow after switching, in a dye that traps magnetic flux but not in a dye of almost similar structure that does not trap magnetic flux. For best results, the magnetic field should be applied before switching and in a direction perpendicular to current flow. The magnetic field effects observed in the dye films at room temperature may be analogous to "anomolous" magnetic phenomena observed in metallic superconductors and/or in superconducting Josephson junctions at low temperatures. The implied possibility of superconductive mechanisms of biomagnetic effects, and apparent relation to the electrical switching seen in wet melanin, give the findings biological relevance.


Quantitative disparities in outpatient antibiotic exposure in a Hungarian county.
Although antibiotic utilization data expressed in defined daily doses (DDDs)/1000 inhabitants/day are often available for a given period and area, the actual antibiotic exposure of the population is rarely explored. We aimed to identify the real antibiotic exposure of the inhabitants of one Hungarian county. The patient-level dispensing data for 2005 for Csongrád County were retrieved from the database of the Hungarian National Health Fund Administration. The number of antibiotic users was quantified, and differences in antibiotic use (quantity and frequency) were explored. Disparities were revealed by Lorenz curves. A new form of Lorenz curve was also introduced. The DDD values for 2005 were used. In 2005, a total of 486 115 antibiotic prescriptions were redeemed, and 3 329 385 DDDs were dispensed to 213 748 different patients; 50.3% of the inhabitants of Csongrád County (total population 424 615) took antibiotics. The average consumption of 1-, 2-, 3-, 4- and 5-time users was 7.5, 14.6, 21.0, 26.9 and 32.2 DDDs per user, respectively. Lorenz curves (including the new form of Lorenz curve) demonstrated the existence of disparities in antibiotic use. One percent of the users with the heaviest consumption were responsible for 6.9% of the total use and redeemed antibiotic prescriptions nine times or more during 2005. Disparities in antibiotic use were detected: half of the inhabitants were exposed to antibiotics, and among antibiotic users, the quantity of antibiotics that they redeemed displayed great variance, mainly due to differences in the prescribing frequency. Special attention should be paid to those with frequent antibiotic use (five times or more annually).


[Feminism and qualitative nursing research].
The purpose of this article was to describe feminism and to propose the integration of a feminist method into qualitative nursing methodology in order to expand the body of nursing knowledge. The world view of feminism including philosophy, epistemology and methodology was outlined, and a feminist grounded theory and feminist ethnography were suggested as a way of strengthening nursing research methodology using literature review. Four different philosophical perspectives of feminism, that is, liberal feminism, radical feminism, Marxist feminism, and social feminism were described. Also epistemological perspectives including feminist empiricism, feminist standpoint, and postmodern feminism, were explained and were related to the methodology and methods of feminism. To enhance the strengths of nursing research within the feminist perspectives, feminist grounded theory and feminist ethnography were exemplified in the paradigm of qualitative nursing research. This paper suggested that incorporation of feminist approaches within nursing is a valuable attempt to expand the body of nursing knowledge and to enhance the quality of nursing care services by rectifying male-oriented knowledge and by empowering women in the care of other people as well as themselves.


Hounsfield units on abdominal computed tomography: a new tool for predicting osteoporosis.
Osteoporosis can cause bone fractures and disability, but early diagnosis faces challenges. Our proposed diagnostic indicators offer a new approach for early detection, which benefits early identification. To determine the most appropriate threshold for predicting osteoporosis in patients with each section of vertebral body. A retrospective analysis of 210 patients, including 646 vertebrae, who had both abdominal computed tomography (CT) and dual-energy X-ray absorptiometry (DXA) within six months. The correlation between DXA T-score and CT Hounsfield units (HU) values was tested by Pearson. The area under the curve (AUC) was calculated using the threshold obtained from the regression equation. The thresholds matching the T-score of -2.5 were 85, 95, 85, and 90 HU for the upper axial plane of the vertebral body (Lau), the middle axial plane of the vertebral body (Lam), the lower axial plane of the vertebral body (Lad), and the mid-sagittal plane of the vertebral body (Lsm), respectively. Defining osteoporosis using CT as Lau ≤ 85, Lam ≤ 95, Lad ≤ 85, or Lsm ≤ 90 HU had a specificity of 88.1% (116/134) and sensitivity of 90.8% (69/76) for distinguishing DXA osteoporosis of the lumbar spine in 210 patients. T-score ≤-2.5 defined as Lau ≤85 or Lam ≤95 or Lad ≤85 or Lsm ≤90 HU had a specificity of 85.9% (275/320) and sensitivity of 82.8% (270/326) for DXA T-score ≤-2.5 in 646 lumbar vertebrae. CT HU values obtained based on different sections of the vertebral body in abdominal CT can be used as a supplementary measure to assess osteoporosis.


The metabolism of phenylmercury by the rat.
The metabolism of [U-(14)C]phenylmercury acetate was studied in the rat. After a single subcutaneous dose a small proportion is excreted unchanged in urine, and a larger amount in bile with some resorption from the gut. The greater part of the dose is broken down in the tissues to yield inorganic mercury which is excreted mainly in faeces, and conjugates of phenol and quinol are excreted in urine. In experiments in vitro phenylmercury is broken down by liver homogenates to release inorganic mercury and benzene; this reaction is effected by the soluble, but not the microsomal, fraction and does not require NADPH or NADH. No elemental mercury is formed under these conditions. It is probable that this reaction occurs in vivo and the benzene produced is rapidly converted into phenol and quinol by microsomal enzymes.


Age-related changes in glutamate release in the CA3 and dentate gyrus of the rat hippocampus.
The present studies employed a novel microelectrode array recording technology to study glutamate release and uptake in the dentate gyrus, CA3 and CA1 hippocampal subregions in anesthetized young, late-middle aged and aged male Fischer 344 rats. The mossy fiber terminals in CA3 showed a significantly decreased amount of KCl-evoked glutamate release in aged rats compared to both young and late-middle-aged rats. Significantly more KCl-evoked glutamate release was seen from perforant path terminals in the DG of late-middle-aged rats compared young and aged rats. The DG of aged rats developed an increased glutamate uptake rate compared to the DG of young animals, indicating a possible age-related change in glutamate regulation to deal with increased glutamate release that occurred in late-middle age. No age-related changes in resting levels of glutamate were observed in the DG, CA3 and CA1. Taken together, these data support dynamic changes to glutamate regulation during aging in subregions of the mammalian hippocampus that are critical for learning and memory.


A subjective assessment of a P300 BCI system for lower-limb rehabilitation purposes.
Recent research has shown that a P300 system can be used while walking without requiring any specific gait-related artifact removal techniques. Also, standard EEG-based Brain-Computer Interfaces (BCI) have not been really assessed for lower limb rehabilitation/prosthesis. Therefore, this paper gives a first baseline estimation (for future BCI comparisons) of the subjective and objective performances of a four-state P300 BCI plus a non-control state for lower-limb rehabilitation purposes. To assess usability and workload, the System Usability Scale and the NASA Task Load Index questionnaires were administered to five healthy subjects after performing a real-time treadmill speed control. Results show that the P300 BCI approach could suit fitness and rehabilitation applications, whereas prosthesis control, which suffers from a low reactivity, appears too sensitive for risky and crowded areas.


[The historical perspective of the sentinel lymph node concept].
The sentinel lymph node procedure is currently standard care in the treatment of breast cancer. The introduction of this procedure in 1992 would not have been possible without the pioneering discoveries regarding lymph nodes and cancer. The Italian surgeon Gaspar Asellius (1581-1626) visualized the lymphatic vessels of a dog after it had been fed and shortly before dissection. At the end of the 17th century, the French anatomists Lauth and Sappey visualized the lymphatics by injecting deceased criminals with mercury. In 1858, the German pathologist Virchow (1821-1902) launched the theory that lymph nodes act as defensive barriers. He also made the first microscopical illustration ofa sentinel lymph node. Gould et al. and Cabaãs independently launched the precursors of the current modern sentinel lymph node concept in 1959 and 1977 respectively. Gould et al. were the first people to use the term "sentinel node'. Cabañas used lymphangiography for the visualisation of the sentinel lymph node. Controversies about the barrier function of the lymph nodes, the fear of skip metastasis and the difficulties of performing the Cabañas procedure, prevented a breakthrough of this concept. In 1992 Morton et al. rediscovered the valuable sentinel node biopsy concept and introduced blue dye for the investigation of patients with melanoma. The combination of lymphoscintigraphy, intra-operative gamma probe guidance and patent blue further increased the reliability of the sentinel lymph node biopsy procedure. Unnecessary lymph gland dissection procedures with considerable morbidity can be prevented by this procedure.


Reactive Reinforcement Learning in Asynchronous Environments.
The relationship between a reinforcement learning (RL) agent and an asynchronous environment is often ignored. Frequently used models of the interaction between an agent and its environment, such as Markov Decision Processes (MDP) or Semi-Markov Decision Processes (SMDP), do not capture the fact that, in an asynchronous environment, the state of the environment may change during computation performed by the agent. In an asynchronous environment, minimizing reaction time-the time it takes for an agent to react to an observation-also minimizes the time in which the state of the environment may change following observation. In many environments, the reaction time of an agent directly impacts task performance by permitting the environment to transition into either an undesirable terminal state or a state where performing the chosen action is inappropriate. We propose a class of reactive reinforcement learning algorithms that address this problem of asynchronous environments by immediately acting after observing new state information. We compare a reactive SARSA learning algorithm with the conventional SARSA learning algorithm on two asynchronous robotic tasks (emergency stopping and impact prevention), and show that the reactive RL algorithm reduces the reaction time of the agent by approximately the duration of the algorithm's learning update. This new class of reactive algorithms may facilitate safer control and faster decision making without any change to standard learning guarantees.


Knock-down of the MEP pathway isogene 1-deoxy-D-xylulose 5-phosphate synthase 2 inhibits formation of arbuscular mycorrhiza-induced apocarotenoids, and abolishes normal expression of mycorrhiza-specific plant marker genes.
The first step of the plastidial methylerythritol phosphate (MEP) pathway is catalyzed by two isoforms of 1-deoxy-D-xylulose 5-phosphate synthase (DXS1 and DXS2). In Medicago truncatula, MtDXS1 and MtDXS2 genes exhibit completely different expression patterns. Most prominently, colonization by arbuscular mycorrhizal (AM) fungi induces the accumulation of certain apocarotenoids (cyclohexenone and mycorradicin derivatives) correlated with the expression of MtDXS2 but not of MtDXS1. To prove a distinct function of DXS2, a selective RNAi approach on MtDXS2 expression was performed in transgenic hairy roots of M. truncatula. Repression of MtDXS2 consistently led to reduced transcript levels in mycorrhizal roots, and to a concomitant reduction of AM-induced apocarotenoid accumulation. The transcript levels of MtDXS1 remained unaltered in RNAi plants, and no phenotypical changes in non-AM plants were observed. Late stages of the AM symbiosis were adversely affected, but only upon strong repression with residual MtDXS2-1 transcript levels remaining below approximately 10%. This condition resulted in a strong decrease in the transcript levels of MtPT4, an AM-specific plant phosphate transporter gene, and in a multitude of other AM-induced plant marker genes, as shown by transcriptome analysis. This was accompanied by an increased proportion of degenerating and dead arbuscules at the expense of mature ones. The data reveal a requirement for DXS2-dependent MEP pathway-based isoprenoid products to sustain mycorrhizal functionality at later stages of the symbiosis. They further validate the concept of a distinct role for DXS2 in secondary metabolism, and offer a novel tool to selectively manipulate the levels of secondary isoprenoids by targeting their precursor supply.


BRCA1/2 germline mutations and their clinical importance in Turkish breast cancer patients.
BRCA1/BRCA2 genes were screened in 117 patients with breast cancer by sequencing. Fourteen percent of patients tested positive for BRCA1/BRCA2 mutations. Four frame shift mutations, four pathogenic missense mutations, and 25 different sequence variations were detected. BRCA mutation positivity was significantly associated with Ki67 (p = .001). BRCA protein expressions were decreased in the patients harboring important mutations and polymorphisms (BRCA1;P508 stop, V1740G, Q1182R, Q1756P and BRCA2;V2466A) related with disease. Our findings contribute significantly to the types of germline BRCA1/BRCA2 mutations and their biological effects in Turkish women. These data could help guide the management of BRCA1/BRCA2 mutation-carrying patients when considering breast-conserving therapy.


Elimination of a group II intron from a plastid gene causes a mutant phenotype.
Group II introns are found in bacteria and cell organelles (plastids, mitochondria) and are thought to represent the evolutionary ancestors of spliceosomal introns. It is generally believed that group II introns are selfish genetic elements that do not have any function. Here, we have scrutinized this assumption by analyzing two group II introns that interrupt a plastid gene (ycf3) involved in photosystem assembly. Using stable transformation of the plastid genome, we have generated mutant plants that lack either intron 1 or intron 2 or both. Interestingly, the deletion of intron 1 caused a strong mutant phenotype. We show that the mutants are deficient in photosystem I and that this deficiency is directly related to impaired ycf3 function. We further show that, upon deletion of intron 1, the splicing of intron 2 is strongly inhibited. Our data demonstrate that (i) the loss of a group II intron is not necessarily phenotypically neutral and (ii) the splicing of one intron can depend on the presence of another.


In vitro biological evaluation of eight different essential oils against Trypanosoma cruzi, with emphasis on Cinnamomum verum essential oil.
Essential oils (EOs) are complex mixtures of secondary metabolites from various plants. It has been shown that several EOs, or their constituents, have inhibitory activity against trypanosomatid protozoa. Thus, we analyzed the biological activity of different EOs on Trypanosoma cruzi, as well as their cytotoxicity on Vero cells. The following EOs were evaluated on T. cruzi epimastigote forms: Cinnamomum verum, Citrus limon, Cymbopogon nardus, Corymbia citriodora, Eucalyptus globulus, Eugenia uniflora, Myrocarpus frondosus, and Rosmarinus officinalis. Inhibitory activity against T. cruzi (IC50/24 h) and cytotoxicity against Vero cells (CC50/24 h) were evaluated by the MTT assay. The EO of C. verum was selected for further evaluation against trypomastigotes and intracellular amastigotes, as well as on parasite metacyclogenesis. Constituents of C. verum EO were identified by GC-MS. One-way ANOVA statistical analysis was performed with GraphPad version 5.01. Cinnamomum verum EO was the most effective against T. cruzi epimastigotes (IC50/24 h = 24.13 μg/ml), followed by Myrocarpus frondosus (IC50/24 h = 60.87 μg/ml) and Eugenia uniflora (IC50/24 h = 70 μg/ml). The EOs of C. citriodora, E. globulus, and R. officinalis showed no activity at concentrations up to 300 μg/ml. Incubation of T. cruzi metacyclic trypomastigotes and intracellular amastigotes with C. verum EO resulted in IC50/24 h values of 5.05 μg/ml and 20 μg/ml, respectively. Therefore, trypomastigotes are more susceptible than epimastigotes, with selectivity index (SI) about 4.7-fold higher (9.78 and 2.05, respectively). Analysis of C. verum EO by GC-MS showed mainly (E)-cinnamaldehyde (81.52%) and eugenol (16.68%). C. verum essential oil is effective against T. cruzi (epimastigotes, trypomastigotes and amastigotes) and interferes with the parasite differentiation process in vitro. Thus, it represents a strong candidate for further studies to improve its activity on pathogenic trypanosomatids.


Synergistic antibacterial effects of localized heat and oxidative stress caused by hydroxyl radicals mediated by graphene/iron oxide-based nanocomposites.
This work develops a composite system of reduced graphene oxide (rGO)-iron oxide nanoparticles (rGO-IONP) that can synergistically induce physical and chemical damage to methicillin-resistant Staphylococcus aureus (MRSA) that are present in subcutaneous abscesses. rGO-IONP was synthesized by the chemical deposition of Fe(2+)/Fe(3+) ions on nanosheets of rGO in aqueous ammonia. The antibacterial efficacy of the as-prepared rGO-IONP was evaluated in a mouse model with MRSA-infected subcutaneous abscesses. Upon exposure to a near-infrared laser in vitro, rGO-IONP synergistically generated localized heat and large amounts of hydroxyl radicals, which inactivated MRSA. The in vivo results reveal that combined treatment with localized heat and oxidative stress that is caused by hydroxyl radicals accelerated the healing of wounds associated with MRSA-infected abscesses. The above results demonstrate that an rGO-IONP nanocomposite system that can effectively inactivate multiple-drug-resistant bacteria in subcutaneous infections was successfully developed. The emergence of methicillin-resistant S. aureus (MRSA) has posed a significant problem in the clinical setting. Thus, it is imperative to develop new treatment strategies against this. In this study, the authors described the use of reduced graphene oxide (rGO)-iron oxide nanoparticles (rGO-IONP) to induce heat and chemical damage to MRSA. This approach may provide a platform the design of other treatment modalities against multiple-drug-resistant bacteria.


Biophysical dose measurement using electron paramagnetic resonance in rodent teeth.
Electron paramagnetic resonance (EPR) dosimetry of human tooth enamel has been widely used in measuring radiation doses in various scenarios. However, there are situations that do not involve a human victim (e.g. tests for suspected environmental overexposures, measurements of doses to experimental animals in radiation biology research, or chronology of archaeological deposits). For such cases we have developed an EPR dosimetry technique making use of enamel of teeth extracted from mice. Tooth enamel from both previously irradiated and unirradiated mice was extracted and cleaned by processing in supersaturated KOH aqueous solution. Teeth from mice with no previous irradiation history exhibited a linear EPR response to the dose in the range from 0.8 to 5.5 Gy. The EPR dose reconstruction for a preliminarily irradiated batch resulted in the radiation dose of (1.4+/-0.2) Gy, which was in a good agreement with the estimated exposure of the teeth. The sensitivity of the EPR response of mouse enamel to gamma radiation was found to be half of that of human tooth enamel. The dosimetric EPR signal of mouse enamel is stable up at least to 42 days after exposure to radiation. Dose reconstruction was only possible with the enamel extracted from molars and premolars and could not be performed with incisors. Electron micrographs showed structural variations in the incisor enamel, possibly explaining the large interfering signal in the non-molar teeth.


A case of a slipped medial rectus muscle after strabismus surgery.
A slipped muscle after strabismus surgery is a well known complication, but only a few cases have been reported in the medical literature. Here, we report a case of a slipped medial rectus muscle that occurred after strabismus surgery for esotropia. The patient was a 10 year old male with an acquired esotropia of 20 PD at distance. For treatment, 3.5 mm bilateral medial rectus recessions was performed. There were no complications during the procedure. On postoperative day 8, the eye position was orthotropic in primary position, but a right exotropia was observed in left gaze. The boy was frequently observed rubbing his right eye after surgery. The right exotropia increased to 30 PD 2 months after surgery. Limited adduction of the right eye, a large widening of the lid fissure on adduction, and mild exophthalmos were observed. Based on the above findings, a slipped medial rectus was suspected, and a second surgery, advancement of the right medial rectus muscle was performed 2.5 months after the initial procedure. After this second procedure, the eyes became orthotropic, and the adduction disorder, large widening the lid fissure, and mild exophthalmos resolved. The slipped muscle may have been caused by insufficient suture and/or excessive rubbing of the eye by the patient. When divergent strabismus is observed after recession of the medial rectus muscle, a slipped muscle should be considered as a differential diagnosis.


The metabolism of sodium cortisone 21-( 35 S)sulphate in the rat.
1. The metabolism of sodium cortisone 21-[(35)S]sulphate was investigated in rats. 2. Quantitative and qualitative experiments showed that substantial amounts of (35)SO(4) (2-) appeared in the urine of free-ranging rats receiving the ester. 3. Whole-body radioautograms indicated considerable biliary elimination of (35)S and also pointed to the liver as the site of metabolism. 4. When female rats with bile-duct cannulae received sodium cortisone 21-[(35)S]sulphate approx. 70% of the dose appeared in the bile as a doubly conjugated steroid (metabolite I). This metabolite was identified as 3alpha-(beta-d-glucopyranuronosido)- 17alpha-hydroxy-21-[(35)S]sulpho-oxy-5alpha-pregnane-11,20-dione. 5. When metabolite I was administered to a rat with a bile-duct cannula 90% of the dose appeared in the bile unchanged. After the administration (intraperitoneally or orally) of metabolite I to free-ranging rats considerable amounts of (35)SO(4) (2-) appeared in the urine. 6. The route by which (35)SO(4) (2-) might be produced from cortisone [(35)S]sulphate in free-ranging animals is discussed.


Induction heating and operator exposure to electromagnetic fields.
Alternating magnetic fields are used in industry for induction heating of metals and semiconductors. Relatively high power, typically of a few to a few hundred of kW is used, and a frequency of operation ranges from 60 Hz to a few tens of MHz. A survey of the magnetic field strengths to which the operators are exposed has shown that these exposures are, in many instances, high compared with recommended exposure limits.


Impact of a timed-release FSH treatment from 2 to 6 months of age in bulls II: Endocrinology, puberty attainment, and mature sperm production in Holstein bulls.
The use of genomic testing in the cattle industries has renewed an interest in hastening bull puberty. In prepubertal males, FSH facilitates Sertoli cell proliferation and testis maturation. The aim of this study was to determine the effect of prepubertal administration of a timed-release FSH (delivered in a hyaluronan solution) on hormone secretion, puberty attainment, and mature sperm production in Holstein bulls in an AI center. Bulls (n = 29) were randomly assigned to one of two treatment groups based on birth date and pedigree. Beginning at 62 days of age (Day 62), bulls were injected im every 3.5 days with either 30 mg FSH (Folltropin-V; NIH-FSH-P1 units) in a 2% hyaluronan solution (FSH-HA, n = 17) or saline (control, n = 12) until Day 170.5. Blood samples to assess FSH, activin A, and testosterone were collected prior to each treatment. Scrotal circumference (SC) and BW were measured monthly. Puberty assessment (ability to ejaculate 5 × 107 sperm, 10% motile) was initiated at Day 244. Average mature daily sperm production (3× wk collection, combined 2 ejaculates) was assessed from Day 571-627. In blood collected every 3.5 days, FSH concentrations within FSH-HA bulls were increased (P < 0.05) over initial Day 62 concentration from Day 93.5-170.5. Concentrations of FSH did not differ between treatments from Day 62-93.5, but were greater (P < 0.05) in FSH-HA than control bulls from Day 97-170.5. Concentrations of activin A assessed for Day 62, 86.5, 107.5, 139, and 170.5 were greater (P < 0.05) in FSH-HA than control bulls on Day 86.5 and 107.5. Treatments did not differ (P > 0.1) in testosterone, BW, or SC. FSH-HA bulls attained puberty at a younger age than control bulls (278 ± 7.7 vs. 303 ± 9.1 days of age, P < 0.05), but mature daily sperm production was not different when measured from Day 571-627 (average 5.84 ± 0.11 billion cells/day, P = 0.5). In summary, FSH administration every 3.5 days from Day 62-170.5 resulted in an increase in FSH concentration beginning at 97 days of age and a hastened age of puberty. We propose this exogenous FSH delivered in hyaluronan initiates a positive feedback loop that includes an increase in activin A production observed on Day 86.5 and 107.5. However, differences in mature sperm production were not realized in this experiment.


Predictive mechanisms linking brain opioids to chronic pain vulnerability and resilience.
Chronic pain is a major global healthcare problem that is currently inadequately treated. In addition, the current use of opioids for treatment has reached far beyond the paucity of evidence for long-term benefits relative to risks. Benefit-risk models for opioid and non-opioid treatments would benefit from a rational, mechanism-based understanding of neuroplastic and neurochemical contributions to chronic pain. Here, we evaluate the findings and limitations of representative research investigating brain neuroplasticity and neurochemistry in chronic pain. In sum, the mechanisms of pain-related neuroplasticity in the brain remain poorly understood because neuroimaging studies have been largely descriptive. We argue that definition is needed of optimal (pain-resilient) and suboptimal (pain-vulnerable) functioning of the endogenous opioid system in order to identify neurochemical contributions to aberrant neuroplasticity in chronic pain. We outline the potential benefits of computational approaches that utilize evolutionary and statistical optimality principles, illustrating this approach with mechanistic hypotheses on opioid function. In particular, we discuss the role of predictive mechanisms in perceptual and associative plasticity and evidence for their modulation by endogenous opioids. Future research should attempt to utilize formal computational models to provide evidence for the clinical validity of this approach, thereby providing a rational basis for future treatment and, ideally, prevention. This article is part of a themed section on Emerging Areas of Opioid Pharmacology. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.14/issuetoc.


Metabolites from the Fungal Endophyte Aspergillus austroafricanus in Axenic Culture and in Fungal-Bacterial Mixed Cultures.
The endophytic fungus Aspergillus austroafricanus isolated from leaves of the aquatic plant Eichhornia crassipes was fermented axenically on solid rice medium as well as in mixed cultures with Bacillus subtilis or with Streptomyces lividans. Chromatographic analysis of EtOAc extract of axenic cultures afforded two new metabolites, namely, the xanthone dimer austradixanthone (1) and the sesquiterpene (+)-austrosene (2), along with five known compounds (3-7). Austradixanthone (1) represents the first highly oxygenated heterodimeric xanthone derivative. When A. austroafricanus was grown in mixed cultures with B. subtilis or with S. lividans, several diphenyl ethers (8-11) including the new austramide (8) were induced up to 29-fold. The structures of new compounds were unambiguously elucidated using 1D- and 2D-NMR spectroscopy, HRESIMS, and chemical derivatization. Compound 7 exhibited weak cytotoxicity against the murine lymphoma L5178Y cell line (EC50 is 12.6 μM). In addition, compounds 9 and 10, which were enhanced in mixed fungal/bacterial cultures, proved to be active against Staphylococcus aureus (ATCC 700699) with minimal inhibitory concentrations (MICs) of 25 μM each (6.6 μg/mL), whereas compound 11 revealed moderate antibacterial activity against B. subtilis 168 trpC2 with an MIC value of 34.8 μM (8 μg/mL).


Medication status and polycystic ovary syndrome in women with bipolar disorder: a preliminary report.
In patients with epilepsy, polycystic ovary (PCO) syndrome has been reported to be associated with the use of the anticonvulsant divalproex sodium. Whether PCO syndrome is associated with divalproex use in patients with bipolar disorder has not previously been explored. Twenty-two female outpatients with a DSM-IV diagnosis of bipolar disorder who were between the ages of 18 and 45 years (inclusive) and who were taking lithium and/or divalproex (10, divalproex monotherapy; 10, lithium monotherapy; 2, divalproex/lithium combination therapy) were evaluated. Patients completed questionnaires about their medical, psychiatric, and reproductive health histories, and body mass indices were calculated. In the early follicular phase of their menstrual cycle, women were examined for hirsutism, given a pelvic ultrasound, and/or assessed for changes in laboratory values such as serum levels of testosterone, free testosterone, estradiol, estrone, dehydroepiandrosterone, dehydroepiandrosterone sulfate, luteinizing hormone, follicle-stimulating hormone, and 17-OH progesterone. All 10 patients on lithium monotherapy, 6 of 10 patients on divalproex monotherapy, and both of the patients on divalproex/lithium combination therapy reported some type of menstrual dysfunction, which, in 4 cases, had preceded the diagnosis of bipolar disorder. Hirsutism was not common in any group, but obesity was prominent in all groups. Ovarian ultrasound revealed an increased number of ovarian follicles in 1 patient taking lithium and in none of the patients taking divalproex. Hormonal screening did not indicate PCO-like changes in any patient. In this pilot study of bipolar patients, PCO-like changes were not seen in women receiving divalproex or lithium. However, independent of therapeutic agent used, the bipolar women in this study reported high rates of menstrual disturbances, suggesting that the hypothalamic-pituitary-gonadal axis may be compromised in some women with bipolar disorder.


Systematic evaluation of multiple NGS platforms for structural variants detection.
Structural variations (SV) are critical genome changes affecting human diseases. Although many hybridization-based methods exist, evaluating SVs through next-generation sequencing (NGS) data is still necessary for broader research exploration. Here, we comprehensively compared the performance of 16 SV callers and multiple NGS platforms using NA12878 whole genome sequencing (WGS) datasets. The results indicated that several SV callers performed well relatively, such as Manta, GRIDSS, LUMPY, TARDIS, FermiKit, and Wham. Meanwhile, all NGS platforms have a similar performance using a single software. Additionally, we found that the source of undetected SVs was mostly from long reads datasets, therefore, the more appropriate strategy for accurate SV detection will be an integration of long and shorter reads in the future. At present, in the period of NGS as a mainstream method in bioinformatics, our study would provide helpful and comprehensive guidelines for specific categories of SV research.


A randomised clinical trial of activity-focussed physiotherapy on patients with distal radius fractures.
Physiotherapy is considered by some authorities to be an important aspect of management in patients following distal radius fractures. There is evidence of improvement in impairment with physiotherapy; however, there is no evidence to support early return of functional activity. Traditional physiotherapy management has focussed on improving impairment; however, there are no trials with emphasis on skill acquisition via motor re-learning principles. Forty-one participants with conservatively managed distal radius fractures were studied in a randomised, single-blinded, prospective trial. Two treatment options were compared: exercise and advice versus activity-focussed physiotherapy with measurement periods of 6 weeks after removal of cast and follow-up at 24 weeks. Participants allocated to the exercise and advice group consulted a physiotherapist an average of 0.9 (SD 0.4) times, while the participants allocated to the more intensive activity-focussed group consulted with physiotherapy a mean of 4.4 (SD 2.3) times. At both 6 and 24 weeks there were no significant differences between the groups for change in impairment (as measured by grip strength, range of motion of wrist flexion and extension and pain intensity), activity limitation and participation restriction, as measured by the Patient-Rated Wrist Evaluation (PRWE). Exercise and advice given by a physiotherapist were equally as effective as activity-focussed physiotherapy in recovery both at 6 and 24 weeks. The results suggest that after removal of cast from fracture of distal radius, patients may routinely require no more than a single session of advice and exercise provided by a physiotherapist.


Facile synthesis of radial-like macroporous superparamagnetic chitosan spheres with in-situ co-precipitation and gelation of ferro-gels.
Macroporous chitosan spheres encapsulating superparamagnetic iron oxide nanoparticles were synthesized by a facile and effective one-step fabrication process. Ferro-gels containing ferrous cations, ferric cations and chitosan were dropped into a sodium hydroxide solution through a syringe pump. In addition, a sodium hydroxide solution was employed for both gelation (chitosan) and co-precipitation (ferrous cations and ferric cations) of the ferro-gels. The results showed that the in-situ co-precipitation of ferro-ions gave rise to a radial morphology with non-spheroid macro pores (large cavities) inside the chitosan spheres. The particle size of iron oxide can be adjusted from 2.5 nm to 5.4 nm by tuning the concentration of the sodium hydroxide solution. Using Fourier Transform Infrared Spectroscopy and X-ray diffraction spectra, the synthesized nanoparticles were illustrated as Fe(3)O(4) nanoparticles. In addition, the prepared macroporous chitosan spheres presented a super-paramagnetic behaviour at room temperature with a saturation magnetization value as high as ca. 18 emu/g. The cytotoxicity was estimated using cell viability by incubating doses (0∼1000 µg/mL) of the macroporous chitosan spheres. The result showed good viability (above 80%) with alginate chitosan particles below 1000 µg/mL, indicating that macroporous chitosan spheres were potentially useful for biomedical applications in the future.


Ovarian cancer detection using optical coherence tomography and convolutional neural networks.
Ovarian cancer has the sixth-largest fatality rate in the United States among all cancers. A non-surgical assay capable of detecting ovarian cancer with acceptable sensitivity and specificity has yet to be developed. However, such a discovery would profoundly impact the pace of the treatment and improvement to patients' quality of life. Achieving such a solution requires high-quality imaging, image processing, and machine learning to support an acceptably robust automated diagnosis. In this work, we propose an automated framework that learns to identify ovarian cancer in transgenic mice from optical coherence tomography (OCT) recordings. Classification is accomplished using a neural network that perceives spatially ordered sequences of tomograms. We present three neural network-based approaches, namely a VGG-supported feed-forward network, a 3D convolutional neural network, and a convolutional LSTM (Long Short-Term Memory) network. Our experimental results show that our models achieve a favorable performance with no manual tuning or feature crafting, despite the challenging noise inherent in OCT images. Specifically, our best performing model, the convolutional LSTM-based neural network, achieves a mean AUC (± standard error) of 0.81 ± 0.037. To the best of the authors' knowledge, no application of machine learning to analyze depth-resolved OCT images of whole ovaries has been documented in the literature. A significant broader impact of this research is the potential transferability of the proposed diagnostic system from transgenic mice to human organs, which would enable medical intervention from early detection of an extremely deadly affliction.


[Selection and acceptability of food].
In this paper the following factors which in one way or the other may have influence on the selection and acceptability of foods are treated: 1. Fisiological and psycological aspects; a) genetic factors, b) neurophysiological factors, c) emotional factors, d) perceptive factors. 2. Physical and ecological aspects. 3. Social and cultural aspects, a) habits and traditions, b) religious believes, c) tabues, d) nutrition faddism, e) prejudice, aversions and perversions, f) social value of foods, g) industrialized foods. 4. Economic aspects. 5. Educational aspects.


Taking blood from children causes no more than minimal harm.
The ethical question of whether taking blood from normal children for research purposes is justified, is determined in part at least, by whether or not the children are harmed. To try to assess the risks, the effects of venepuncture on a group of healthy subjects were studied, by means of a parental questionnaire completed approximately eighteen months after the venepuncture had taken place. Ninety-two healthy children aged between 6 and 8 had a blood sample taken for non-therapeutic reasons as part of a research study. Questionnaire responses reveal few negative effects, and in some cases positive effects.


alpha-DNA. VI: Comparative study of alpha- and beta-anomeric oligodeoxyribonucleotides in hybridization to mRNA and in cell free translation inhibition.
alpha and beta-anomeric d(G2T12G2) oligodeoxyribonucleotides were compared for their hybridization to rA12: the observed melting temperatures are 27 degrees C for beta-oligodeoxyribonucleotide/RNA hybrid and 53 degrees C for alpha-oligodeoxyribonucleotide/RNA. alpha-oligonucleotides with the four bases, complementary to natural mRNAs, were synthesized for the first time, labeled at their 5'-end with [32P] and used as probes in Northern blot experiments. In spite of these higher affinities for their target RNA's, they were unable to block translation of natural or synthetic mRNA's in rabbit reticulocyte lysate. We have studied the RNase H activity on model rA12:alpha- or beta-d(G2T12G2) hybrids or on mRNA:alpha- or beta-oligonucleotides hybrids. Specific hybridization protects RNA strech when using alpha-oligonucleotides but not beta-oligonucleotides. Thus, our results show the inability of RNase H to degrade RNA in alpha-oligodeoxyribonucleotides:RNA duplexes.


Population ethics and the prospects for fertility policy as climate mitigation policy.
What are the prospects for using population policy as tool to reduce carbon emissions? In this paper, we review evidence from population science, in order to inform debates in population ethics that, so far, have largely taken place within the academic philosophy literature. In particular, we ask whether fertility policy is likely to have a large effect on carbon emissions, and therefore on temperature change. Our answer is no. Prospects for a policy of fertility-reduction-as-climate-mitigation are limited by population momentum, a demographic factor that limits possible variation in the size of the population, even if fertility rates change very quickly. In particular, a hypothetical policy that instantaneously changed fertility and mortality rates to replacement levels would nevertheless result in a population of over 9 billion people in 2060. We use a leading climate-economy model to project the consequence of such a hypothetical policy for climate change. As a standalone mitigation policy, such a hypothetical change in the size of the future population - much too large to be implementable by any foreseeable government program - would reduce peak temperature change only to 6.4°C, relative to 7.1°C under the most likely population path. Therefore, fertility reduction is unlikely to be an adequate core approach to climate mitigation.


Demystifying the association between economic development, transportation, tourism, renewable energy, and ecological footprint in Bay of Bengal Initiative for Multi-Sectoral Technical and Economic Cooperation region during globalization mode.
The Bay of Bengal Initiative for Multi-Sectoral Technical and Economic Cooperation (BIMSTEC) economies have yet to meaningfully contribute to accomplishing Sustainable Development Goals (SDG 7) affordable and clean energy, (SDG 8) decent work and economic growth, and (SDG 13) climate action. Dealing with this issue might require a shift or alteration of policy framework that is the major theme of this study. Consequently, this present research inspects the influence of economic growth, transportation, tourism sector development, and renewable energy on ecological footprint using panel time series from 1990 and 2019 for the BIMSTEC region. To evaluate this dynamic nexus between the mentioned environmental pollution drivers of ecological footprint, this study employed the augumented mean group (AMG) and common correlated effect mean group (CCEMG) regression estimators after detection of cross-sectional dependency. The empirical outcomes denote that economic growth and transportation sector of BIMSTEC countries increase the levels of ecological footprint. Conversely, tourism sector development, globalization, and renewable energy protect the ecological excellence in the region. Moreover, it is observed that a unidirectional causality exists from economic growth to ecological footprint, ecological footprint to transportation, tourism to ecological footprint, and globalization to ecological footprint, while bidirectional causality exists between renewable energy and ecological footprint. By observing the positive function of tourism, green energy, and globalization on sustainable environment progress, central authorities are capable to redesign policies concerning supportable efficient technologies and regulate globalization towards green programs and agenda to reduce global warming.


Infrared neural stimulation in the cochlea.
The application of photonics to manipulate and stimulate neurons and to study neural networks has gained momentum over the last decade. Two general methods have been used: the genetic expression of light or temperature sensitive ion channels in the plasma membrane of neurons (Optogenetics and Thermogenetics) and the direct stimulation of neurons using infrared radiation (Infrared Neural Stimulation, INS). Both approaches have their strengths and challenges, which are well understood with a profound understanding of the light tissue interaction(s). This paper compares the opportunities of the methods for the use in cochlear prostheses. Ample data are already available on the stimulation of the cochlea with INS. The data show that the stimulation is selective, feasible at rates that would be sufficient to encode acoustic information and may be beneficial over conventional pulsed electrical stimulation. A third approach, using lasers in stress confinement to generate pressure waves and to stimulate the functional cochlea mechanically will also be discussed.


Differences in Reactions to Media Reporting of a Celebrity Suicide Between Emotionally Distressed and Nondistressed Individuals in Taiwan.
Background: The differences in emotional reactions toward media reports of celebrity suicides between distressed and non-distressed individuals have rarely been explored. Aim: We aimed to explore differences in reaction patterns to the news coverage of a celebrity's suicide between distressed and nondistressed individuals in Taiwan. Method: We conducted an Internet survey to assess the impact of the news coverage of a celebrity's suicide. A series of χ2 tests, Spearman's rank correlations, and logistic regression analyses were performed to compare the differences in emotional involvement and psychological impact after exposure to suicide news between distressed and nondistressed individuals. Results: A total of 1,258 Internet users responded to the survey (236 men and 1,020 women). Compared with nondistressed individuals, distressed individuals (n = 537) were more likely to feel suicidal (11% vs. 2%, p < .001) after reading the news. They were less willing to seek help when in need (62% vs. 77%, p < .001). All the variables remained significant after adjustment for sociodemographic variables in regression analyses. Limitations: The Internet-based survey may suffer from selection bias. Conclusion: Extensive media reporting of suicide stories has a profound impact on readers, particularly on vulnerable groups. Responsible reporting of suicide incidents is crucial for suicide prevention.


Metachronous manifestations of Sweet's syndrome in a neutropenic patient with Fanconi anemia.
Sweet's syndrome (SS) is an acute, febrile neutrophilic dermatosis that frequently presents with leukocytosis and erythematous plaques. Lesions show neutrophilic infiltration of the dermis and rarely other organs. We report the case of an adolescent male with chronic pancytopenia secondary to Fanconi anemia (FA) who presented with acute respiratory distress. Despite an exhaustive and ultimately unrevealing work-up, the diagnosis of pulmonary SS was not made until he developed characteristic cutaneous lesions 4 months later. Comprehensive review of pathological specimens revealed metachronous SS manifestations with infiltrates in lung parenchyma, dermis, and subcutis in this neutropenic patient.


'Complex' but coping: experience of symptoms of tuberculosis and health care seeking behaviours--a qualitative interview study of urban risk groups, London, UK.
Tuberculosis awareness, grounded in social cognition models of health care seeking behaviour, relies on the ability of individuals to recognise symptoms, assess their risk and access health care (passive case finding). There is scant published research into the health actions of 'hard-to-reach' groups with tuberculosis, who represent approximately 17% of the London TB caseload. This study aimed to analyse patients' knowledge of tuberculosis, their experiences of symptoms and their health care seeking behaviours. Qualitative interviews were conducted with 17 participants, predominantly homeless and attending a major tuberculosis centre in London, UK. Most had complex medical and social needs including drug and alcohol use or immigration problems affecting entitlement to social welfare. Analytical frameworks aimed to reflect the role of broader social structures in shaping individual health actions. Although participants demonstrated some knowledge of tuberculosis their awareness of personal risk was low. Symptoms commonly associated with tuberculosis were either not recognised or were attributed to other causes for which participants would not ordinarily seek health care. Many accessed health care by chance and, for some, for health concerns other than tuberculosis. Health education, based on increasing awareness of symptoms, may play a limited role in tuberculosis care for populations with complex health and social needs. The findings support the intensification of outreach initiatives to identify groups at risk of tuberculosis and the development of structured care pathways which support people into prompt diagnosis and treatment.


A single amino acid change within the R2 domain of the VvMYB5b transcription factor modulates affinity for protein partners and target promoters selectivity.
Flavonoid pathway is spatially and temporally controlled during plant development and the transcriptional regulation of the structural genes is mostly orchestrated by a ternary protein complex that involves three classes of transcription factors (R2-R3-MYB, bHLH and WDR). In grapevine (Vitis vinifera L.), several MYB transcription factors have been identified but the interactions with their putative bHLH partners to regulate specific branches of the flavonoid pathway are still poorly understood. In this work, we describe the effects of a single amino acid substitution (R69L) located in the R2 domain of VvMYB5b and predicted to affect the formation of a salt bridge within the protein. The activity of the mutated protein (name VvMYB5b(L), the native protein being referred as VvMYB5b(R)) was assessed in different in vivo systems: yeast, grape cell suspensions, and tobacco. In the first two systems, VvMYB5b(L) exhibited a modified trans-activation capability. Moreover, using yeast two-hybrid assay, we demonstrated that modification of VvMYB5b transcriptional properties impaired its ability to correctly interact with VvMYC1, a grape bHLH protein. These results were further substantiated by overexpression of VvMYB5b(R) and VvMYB5b(L) genes in tobacco. Flowers from 35S::VvMYB5b(L) transgenic plants showed a distinct phenotype in comparison with 35S::VvMYB5b(R) and the control plants. Finally, significant differences in transcript abundance of flavonoid metabolism genes were observed along with variations in pigments accumulation. Taken together, our findings indicate that VvMYB5b(L) is still able to bind DNA but the structural consequences linked to the mutation affect the capacity of the protein to activate the transcription of some flavonoid genes by modifying the interaction with its co-partner(s). In addition, this study underlines the importance of an internal salt bridge for protein conformation and thus for the establishment of protein-protein interactions between MYB and bHLH transcription factors. Mechanisms underlying these interactions are discussed and a model is proposed to explain the transcriptional activity of VvMYB5(L) observed in the tobacco model.


Wolbachia induces male-specific mortality in the mosquito Culex pipiens (LIN strain).
Wolbachia are maternally inherited endosymbionts that infect a diverse range of invertebrates, including insects, arachnids, crustaceans and filarial nematodes. Wolbachia are responsible for causing diverse reproductive alterations in their invertebrate hosts that maximize their transmission to the next generation. Evolutionary theory suggests that due to maternal inheritance, Wolbachia should evolve toward mutualism in infected females, but strict maternal inheritance means there is no corresponding force to select for Wolbachia strains that are mutualistic in males. Using cohort life-table analysis, we demonstrate that in the mosquito Culex pipiens (LIN strain), Wolbachia-infected females show no fitness costs due to infection. However, Wolbachia induces up to a 30% reduction in male lifespan. These results indicate that the Wolbachia infection of the Culex pipiens LIN strain is virulent in a sex-specific manner. Under laboratory situations where mosquitoes generally mate at young ages, Wolbachia strains that reduce male survival could evolve by drift because increased mortality in older males is not a significant selective force.


An attenuated projector-backprojector for iterative SPECT reconstruction.
A new ray-driven projector-backprojector which can easily be adapted for hardware implementation is described and simulated in software. The projector-backprojector discretely models the attenuated Radon transform of a source distributed within an attenuating medium as line integrals of discrete pixels, obtained using the standard sampling technique of averaging the emission source or attenuation distribution over small square regions. Attenuation factors are calculated for each pixel during the projection and backprojection operations instead of using precalculated values. The calculation of the factors requires a specification of the attenuation distribution, estimated either from an assumed constant distribution and an approximate body outline or from transmission measurements. The distribution of attenuation coefficients is stored in memory for efficient access during the projection and backprojection operations. The reconstruction of the source distribution is obtained by using a conjugate gradient or SIRT type iterative algorithm which requires one projection and one backprojection operation for each iteration.


Are public-private partnerships a healthy option? A systematic literature review.
Governments around the world, but especially in Europe, have increasingly used private sector involvement in developing, financing and providing public health infrastructure and service delivery through public-private partnerships (PPPs). Reasons for this uptake are manifold ranging from rising expenditures for refurbishing, maintaining and operating public assets, and increasing constraints on government budgets stifle, seeking innovation through private sector acumen and aiming for better risk management. Although PPPs have attracted practitioner and academic interest over the last two decades, there has been no attempt to integrate the general and health management literature to provide a holistic view of PPPs in healthcare delivery. This study analyzes over 1400 publications from a wide range of disciplines over a 20-year time period. We find that despite the scale and significance of the phenomenon, there is relatively limited conceptualization and in-depth empirical investigation. Based on bibliographic and content analyses, we synthesize formerly dispersed research perspectives into a comprehensive multi-dimensional framework of public-private partnerships. In so doing, we provide new directions for further research and practice.


Response to a large rotavirus outbreak on South Tarawa, Kiribati, 2013.
In July 2013, during annual independence celebrations in Kiribati, staff at Tungaru Central Hospital on South Tarawa reported an increase in children presenting with severe diarrhoea. This report describes the outbreak investigation, findings and response. After notification of the outbreak, all health facilities on South Tarawa began reporting cases of acute diarrhoea and/or vomiting through the early warning syndromic surveillance system on a daily basis. Community awareness was raised and the public was encouraged to present to a health facility if ill with acute gastroenteritis. Specimens were collected and sent for laboratory testing. Between 10 and 24 July 2013, 1118 cases of gastroenteritis were reported; 103 were hospitalized and six died. The median age of cases was one year (range: 0-68 years); 93.4% were aged less than five years. Rotavirus was identified in 81% of specimens tested. The outbreak response included enhanced surveillance, community education, clinical training and changes to in-hospital patient management for infection control. This outbreak was the largest diarrhoea outbreak in Kiribati in five years. Factors that may have contributed to the magnitude and severity of the outbreak included high household density, inadequate sanitation infrastructure and a mass gathering--all increasing the chance of transmission--as well as limited clinical response capacity. The current outbreak highlights the importance of clinical management to minimize severe dehydration and death. Rotavirus vaccination should be considered as an adjunct to other comprehensive enteric disease control measures as recommended by the World Health Organization.


Kinetic serum creatinine assays I. The role of various factors in determining specificity.
We studied the specificity of kinetic methods of analysis, with emphasis on creatinine determination. The error contributed by an interferent that reacts with the analytical reagent and absorbs at the wavelength of analysis is shown to be related to the ratio of the equilibrium absorbances of the analyte and interferent, the ratio of the rate constants, the extent of the analytical reaction, and the duration of the measurement. To assess the potential diminution of the interference with kinetic methods, we determined the rate constants of several alpha-keto acids in the Jaffé reaction. Evaluation of the advantages of various measuring techniques with respect to specificity showed the main factors to be the extent of the analytical reaction and the relative value of the rate constants. Total specificity for creatine appears to be unattainable with currently used kinetic techniques.


Quantum correlations and secret bits.
It is shown that (i) all entangled states can be mapped by single-copy measurements into probability distributions containing secret correlations, and (ii) if a probability distribution obtained from a quantum state contains secret correlations, then this state has to be entangled. These results prove the existence of a two-way connection between secret and quantum correlations in the process of preparation. They also imply that either it is possible to map any bound entangled state into a distillable probability distribution or bipartite bound information exists.