Evaluating Transcription Factor Activity Changes by Scoring Unexplained Target Genes in Expression Data.
Several methods predict activity changes of transcription factors (TFs) from a given regulatory network and measured expression data. But available gene regulatory networks are incomplete and contain many condition-dependent regulations that are not relevant for the specific expression measurement. It is not known which combination of active TFs is needed to cause a change in the expression of a target gene. A method to systematically evaluate the inferred activity changes is missing. We present such an evaluation strategy that indicates for how many target genes the observed expression changes can be explained by a given set of active TFs. To overcome the problem that the exact combination of active TFs needed to activate a gene is typically not known, we assume a gene to be explained if there exists any combination for which the predicted active TFs can possibly explain the observed change of the gene. We introduce the i-score (inconsistency score), which quantifies how many genes could not be explained by the set of activity changes of TFs. We observe that, even for these minimal requirements, published methods yield many unexplained target genes, i.e. large i-scores. This holds for all methods and all expression datasets we evaluated. We provide new optimization methods to calculate the best possible (minimal) i-score given the network and measured expression data. The evaluation of this optimized i-score on a large data compendium yields many unexplained target genes for almost every case. This indicates that currently available regulatory networks are still far from being complete. Both the presented Act-SAT and Act-A* methods produce optimal sets of TF activity changes, which can be used to investigate the difficult interplay of expression and network data. A web server and a command line tool to calculate our i-score and to find the active TFs associated with the minimal i-score is available from https://services.bio.ifi.lmu.de/i-score.


CARD9 is a novel caspase recruitment domain-containing protein that interacts with BCL10/CLAP and activates NF-kappa B.
BCL10/CLAP is an activator of apoptosis and NF-kappaB signaling pathways and has been implicated in B cell lymphomas of mucosa-associated lymphoid tissue. Although its role in apoptosis remains to be determined, BCL10 likely activates NF-kappaB through the IKK complex in response to upstream stimuli. The N-terminal caspase recruitment domain (CARD) of BCL10 has been proposed to function as an activation domain that mediates homophilic interactions with an upstream CARD-containing NF-kappaB activator. To identify upstream signaling partners of BCL10, we performed a mammalian two-hybrid analysis and identified CARD9 as a novel CARD-containing protein that interacts selectively with the CARD activation domain of BCL10. When expressed in cells, CARD9 binds to BCL10 and activates NF-kappaB. Furthermore, endogenous CARD9 is found associated with BCL10 suggesting that both proteins form a pre-existing signaling complex within cells. CARD9 also self-associates and contains extensive coiled-coil motifs that may function as oligomerization domains. We propose here that CARD9 is an upstream activator of BCL10 and NF-kappaB signaling.


Electrical brain responses in language-impaired children reveal grammar-specific deficits.
Scientific and public fascination with human language have included intensive scrutiny of language disorders as a new window onto the biological foundations of language and its evolutionary origins. Specific language impairment (SLI), which affects over 7% of children, is one such disorder. SLI has received robust scientific attention, in part because of its recent linkage to a specific gene and loci on chromosomes and in part because of the prevailing question regarding the scope of its language impairment: Does the disorder impact the general ability to segment and process language or a specific ability to compute grammar? Here we provide novel electrophysiological data showing a domain-specific deficit within the grammar of language that has been hitherto undetectable through behavioural data alone. We presented participants with Grammatical(G)-SLI, age-matched controls, and younger child and adult controls, with questions containing syntactic violations and sentences containing semantic violations. Electrophysiological brain responses revealed a selective impairment to only neural circuitry that is specific to grammatical processing in G-SLI. Furthermore, the participants with G-SLI appeared to be partially compensating for their syntactic deficit by using neural circuitry associated with semantic processing and all non-grammar-specific and low-level auditory neural responses were normal. The findings indicate that grammatical neural circuitry underlying language is a developmentally unique system in the functional architecture of the brain, and this complex higher cognitive system can be selectively impaired. The findings advance fundamental understanding about how cognitive systems develop and all human language is represented and processed in the brain.


Association study of trauma load and SLC6A4 promoter polymorphism in posttraumatic stress disorder: evidence from survivors of the Rwandan genocide.
As exposure to different types of traumatic stressors increases, the occurrence of posttraumatic stress disorder (PTSD) increases. However, because some people exhibit either surprising resilience or high vulnerability, further influencing factors have been conjectured, such as gene-environment interactions. The SLC6A4 gene, which encodes serotonin transporter, has been identified as predisposing toward differential emotional processing between genotypes of its promoter polymorphism. We investigated 408 refugees from the Rwandan genocide and assessed lifetime exposure to traumatic events, PTSD (according to DSM-IV) status, and genotype of the SLC6A4 promoter polymorphism. The study was conducted from March 2006 to February 2007. The prevalence of PTSD approached 100% when traumatic exposure reached extreme levels. However, persons homozygous for the short allele of the SLC6A4 promoter polymorphism showed no dose-response relationship but were at high risk for developing PTSD after very few traumatic events. This genotype influence vanished with increasing exposure to traumatic stressors. We find evidence for a gene-environment interplay for PTSD and show that genetic influences lose importance when environmental factors cause an extremely high trauma burden to an individual. In the future, it may be important to determine whether the effectiveness of therapeutic interventions in PTSD is also modulated by the SLC6A4 genotype.


THERMAL DENATURATION OF COLLAGEN IN THE DISPERSED AND SOLID STATE.
Thermal denaturation temperature of newly reconstituted collagen fibrils from rat tail tendons is 52 degrees C compared with 42 degrees C for neutral solutions. This suggests that the increase in concentration of collagen within the fibril increases the stability of the individual molecules. The absence of firm intermolecular bonds in these fibrils rules out crosslinking as an explanation for increased stability. "Aging" at 37 degrees C up to 1 year raises the shrinkage temperature of reconstituted fibrous gels by 4 degrees to 6 degrees C and greatly increases resistance to dissolution at high temperature. The newly formed fibrils dissolve without shrinking, whereas older gels exhibit shrinkage before dissolution. Since nearly all extractable collagen is in the form of fibrillar aggregates in tissue, it is unlikely that thermal denaturation occurs at body temperature; therefore it could not be involved as a necessary stage in collagen resorption.


Dual role of TFIIH in DNA excision repair and in transcription by RNA polymerase II.
The RNA polymerase II general transcription factor TFIIH is composed of several polypeptides. The observation that the largest subunit of TFIIH is the excision-repair protein XPB/ERCC3 (ref. 1), a helicase implicated in the human DNA-repair disorders xeroderma pigmentosum (XP) and Cockayne's syndrome, suggests a functional link between transcription and DNA repair. To understand the connection between these two cellular processes, we have extensively purified and functionally analysed TFIIH. We find that TFIIH has a dual role, being required for basal transcription of class II genes and for participation in DNA-excision repair. TFIIH is shown to complement three different cell extracts deficient in excision repair: XPB/ERCC3, XPC and XPD/ERCC2. The complementation of XPB and XPD is a consequence of ERCC3 and ERCC2 being integral subunits of TFIIH, whereas complementation of XPC is due to an association of this polypeptide with TFIIH. We found that the general transcription factor IIE negatively modulates the helicase activity of TFIIH through a direct interaction between TFIIE and the ERCC3 subunit of TFIIH.


Rapid genotype imputation from sequence without reference panels.
Inexpensive genotyping methods are essential for genetic studies requiring large sample sizes. In human studies, array-based microarrays and high-density haplotype reference panels allow efficient genotype imputation for this purpose. However, these resources are typically unavailable in non-human settings. Here we describe a method (STITCH) for imputation based only on sequencing read data, without requiring additional reference panels or array data. We demonstrate its applicability even in settings of extremely low sequencing coverage, by accurately imputing 5.7 million SNPs at a mean r(2) value of 0.98 in 2,073 outbred laboratory mice (0.15× sequencing coverage). In a sample of 11,670 Han Chinese (1.7× coverage), we achieve accuracy similar to that of alternative approaches that require a reference panel, demonstrating that our approach can work for genetically diverse populations. Our method enables straightforward progression from low-coverage sequence to imputed genotypes, overcoming barriers that at present restrict the application of genome-wide association study technology outside humans.


Visual and Semantic Knowledge Transfer for Large Scale Semi-Supervised Object Detection.
Deep CNN-based object detection systems have achieved remarkable success on several large-scale object detection benchmarks. However, training such detectors requires a large number of labeled bounding boxes, which are more difficult to obtain than image-level annotations. Previous work addresses this issue by transforming image-level classifiers into object detectors. This is done by modeling the differences between the two on categories with both image-level and bounding box annotations, and transferring this information to convert classifiers to detectors for categories without bounding box annotations. We improve this previous work by incorporating knowledge about object similarities from visual and semantic domains during the transfer process. The intuition behind our proposed method is that visually and semantically similar categories should exhibit more common transferable properties than dissimilar categories, e.g. a better detector would result by transforming the differences between a dog classifier and a dog detector onto the cat class, than would by transforming from the violin class. Experimental results on the challenging ILSVRC2013 detection dataset demonstrate that each of our proposed object similarity based knowledge transfer methods outperforms the baseline methods. We found strong evidence that visual similarity and semantic relatedness are complementary for the task, and when combined notably improve detection, achieving state-of-the-art detection performance in a semi-supervised setting.


[Molluscacidal activity of Paraiso (Melia azedarach L.) (Meliaceae) on Lymnaea cubensis, host snail of fascioliasis].
Fasciolosis is a source of serious economic loss in various regions of Brazil and when control measures of its host (molluscs) are not taken under favorable ecological conditions, isolated cases of human Fasciolosis may occur. Among the alternative measures for its control is the use of vegetable extracts and the purpose of this project is the assessment of the use of juice extracted from the fruit and seeds of the Paraiso plant (Melia azedarach L.) in the control of Lymnaea cubensis, the main vector of Fasciolosis in Cuba. Various concentrations of the juice extracted from the Paraiso fruit (Melia azedarach L.) were tested to determine the average and maximum lethal doses (DL50 and DL90' respectively) used in a computerized PROBIT-LOG program. Seven experimental series were undertaken, 72 molluscs being used in each of them. Three groups of ten molluscs were tested to determine the effect on cardiac frequency, two of them being treated with CL50 = 0.88627 and CL90 = 1.7641, respectively, the third being used as control. A considerable effect of both doses on the cardiac frequency of the mollusc studied was observed. The results were encouraging, for they demonstrated the potential use of this plant in the control of undesirable molluscs.


A 9-year follow-up of a girl with pyridoxine (vitamin B6)-dependent seizures: magnetic resonance spectroscopy findings.
Being an extremely rare condition makes the diagnosis of pyridoxine-dependent seizures (PDS) difficult. Early diagnosis of PDS is very important to prevent unwanted clinical outcomes. Elevated levels of glutamate and decreased levels of y-aminobutyric acid (GABA) in the frontal and parietal cortices are detected in this disorder. Here we present an 18 year old girl with PDS, who was reported 9 years ago with Magnetic Resonance Spectroscopy (MRS) findings. Present and past MRS findings showed a decrease in N-acetyl-aspartate-to creatine ratio on MRS. In this case it is surprising that neuronal damage has been preceded despite the administration of accurate treatment. That can be because of delay in treatment and/or under treatment.


Optic nerve head and retinal nerve fiber layer analysis: a report by the American Academy of Ophthalmology.
To evaluate the current published literature on the use of optic nerve head (ONH) and retinal nerve fiber layer (RNFL) measurement devices in diagnosing open-angle glaucoma and detecting progression. A search of peer-reviewed literature was conducted on February 15, 2006 in PubMed and the Cochrane Library for the period January 2003 to February 2006. The search was limited to studies of adults in English-language journals and yielded 442 citations. The panel reviewed the abstracts of these articles and selected 159 articles of possible clinical relevance for review. Of these 159 full-text articles, 82 were determined to be relevant for the first author and methodologist to review and rate according to the quality of evidence. There were no studies classified as having the highest level of evidence (level I). The ONH and RNFL imaging instruments reviewed in this assessment were determined to be highly effective in distinguishing eyes with glaucomatous visual field (VF) loss from normal eyes without VF loss, based on level II evidence. In addition, some studies demonstrated that parameters from ONH or RNFL imaging predicted the development of VF defects among glaucoma suspects. Studies on detecting glaucoma progression showed that although there was often agreement on progression between the structural and functional (VF) tests, a significant proportion of glaucoma patients progressed by either the structural or the functional test alone. The ONH and RNFL imaging devices provide quantitative information for the clinician. Based on studies that have compared the various available technologies directly, there is no single imaging device that outperforms the others in distinguishing patients with glaucoma from controls. Ongoing advances in imaging and related software, as well as the impracticalities associated with obtaining and assessing optic nerve stereophotographs, have made imaging increasingly important in many practice settings. The information obtained from imaging devices is useful in clinical practice when analyzed in conjunction with other relevant parameters that define glaucoma diagnosis and progression.


Lymphatic Pump Treatment Mobilizes Bioactive Lymph That Suppresses Macrophage Activity In Vitro.
By promoting the recirculation of tissue fluid, the lymphatic system preserves tissue health, aids in the absorption of gastrointestinal lipids, and supports immune surveillance. Failure of the lymphatic system has been implicated in the pathogenesis of several infectious and inflammatory diseases. Thus, interventions that enhance lymphatic circulation, such as osteopathic lymphatic pump treatment (LPT), should aid in the management of these diseases. To determine whether thoracic duct lymph (TDL) mobilized during LPT would alter the function of macrophages in vitro. The thoracic ducts of 6 mongrel dogs were cannulated, and TDL samples were collected before (baseline), during, and 10 minutes after LPT. Thoracic duct lymph flow was measured, and TDL samples were analyzed for protein concentration. To measure the effect of TDL on macrophage activity, RAW 264.7 macrophages were cultured for 1 hour to acclimate. After 1 hour, cell-free TDL collected at baseline, during LPT, and after TDL was added at 5% total volume per well and co-cultured with or without 500 ng per well of lipopolysaccharide (LPS) for 24 hours. As a control for the addition of 5% TDL, macrophages were cultured with phosphate-buffered saline (PBS) at 5% total volume per well and co-cultured with or without 500 ng per well of LPS for 24 hours. After culture, cell-free supernatants were assayed for nitrite (NO2-), tumor necrosis factor α (TNF-α) and interleukin 10 (IL-10). Macrophage viability was measured using flow cytometry. Lymphatic pump treatment significantly increased TDL flow and the flux of protein in TDL (P<.001). After culture, macrophage viability was approximately 90%. During activation with LPS, baseline TDL, TDL during LPT, and TDL after LPT significantly decreased the production of NO2-, TNF-α, and IL-10 by macrophages (P<.05). However, no significant differences were found in viability or the production of NO2-, TNF-α, or IL-10 between macrophages cultured with LPS plus TDL taken before, during, and after LPT (P>.05). The redistribution of protective lymph during LPT may provide scientific rationale for the clinical use of LPT to reduce inflammation and manage edema.


Hyperspectral confocal microscope.
We have developed a new, high performance, hyperspectral microscope for biological and other applications. For each voxel within a three-dimensional specimen, the microscope simultaneously records the emission spectrum from 500 nm to 800 nm, with better than 3 nm spectral resolution. The microscope features a fully confocal design to ensure high spatial resolution and high quality optical sectioning. Optical throughput and detection efficiency are maximized through the use of a custom prism spectrometer and a backside thinned electron multiplying charge coupled device (EMCCD) array. A custom readout mode and synchronization scheme enable 512-point spectra to be recorded at a rate of 8300 spectra per second. In addition, the EMCCD readout mode eliminates curvature and keystone artifacts that often plague spectral imaging systems. The architecture of the new microscope is described in detail, and hyperspectral images from several specimens are presented.


Perceived Stress and Coffee and Energy Drink Consumption Predict Poor Sleep Quality in Podiatric Medical Students A Cross-sectional Study.
A cross-sectional survey administered to first- and second-year podiatric medical students aimed to investigate the effect of coffee intake, energy drink consumption, and perceived stress on sleep quality in medical students during their preclinical studies. Ninety-eight of 183 students contacted (53.6%) completed a questionnaire comprising standard instruments measuring sleep quality (Pittsburgh Sleep Quality Index), daytime sleepiness (Epworth Sleepiness scale), and perceived stress (ten-item Perceived Stress Scale). Furthermore, we investigated coffee and energy drink consumption. Logistic regression was conducted to identify factors associated with poor sleep quality and the relation between sleep quality and academic performance (grade point average). High prevalences of poor sleep quality, excessive daytime sleepiness, and perceived stress were reported. In addition, higher odds of developing poor sleep quality were associated with coffee and energy drink intake, perceived stress, and excessive daytime sleepiness. The total Pittsburgh Sleep Quality Index score was inversely correlated with grade point average. First- and second-year podiatric medical students have poor sleep quality. Further research is needed to identify effective strategies to reduce stress and decrease coffee and energy drink intake to minimize their negative effect on sleep quality and academic performance in podiatric medical students.


Comparison of imatinib 400 mg and 800 mg daily in the front-line treatment of high-risk, Philadelphia-positive chronic myeloid leukemia: a European LeukemiaNet Study.
Imatinib mesylate (IM), 400 mg daily, is the standard treatment of Philadelphia-positive (Ph(+)) chronic myeloid leukemia (CML). Preclinical data and results of single-arm studies raised the suggestion that better results could be achieved with a higher dose. To investigate whether the systematic use of a higher dose of IM could lead to better results, 216 patients with Ph(+) CML at high risk (HR) according to the Sokal index were randomly assigned to receive IM 800 mg or 400 mg daily, as front-line therapy, for at least 1 year. The CCgR rate at 1 year was 64% and 58% for the high-dose arm and for the standard-dose arm, respectively (P = .435). No differences were detectable in the CgR at 3 and 6 months, in the molecular response rate at any time, as well as in the rate of other events. Twenty-four (94%) of 25 patients who could tolerate the full 800-mg dose achieved a CCgR, and only 4 (23%) of 17 patients who could tolerate less than 350 mg achieved a CCgR. This study does not support the extensive use of high-dose IM (800 mg daily) front-line in all CML HR patients. This trial was registered at www.clinicaltrials.gov as #NCT00514488.


How are molecular crowding and the spatial organization of a biopolymer interrelated.
In a crowded cellular interior, dissolved biomolecules or crowders exert excluded volume effects on other biomolecules, which in turn control various processes including protein aggregation and chromosome organization. As a result of these effects, a long chain molecule can be phase-separated into a condensed state, redistributing the surrounding crowders. Using computer simulations and a theoretical approach, we study the interrelationship between molecular crowding and chain organization. In a parameter space of biological relevance, the distributions of monomers and crowders follow a simple relationship: the sum of their volume fractions rescaled by their size remains constant. Beyond a physical picture of molecular crowding it offers, this finding explains a few key features of what has been known about chromosome organization in an E. coli cell.


Probing morphological changes in polymersomes with magnetic birefringence.
Magnetic birefringence was used for in situ monitoring of the morphological changes in diamagnetic polymersomes during shape-transformation by dialysis. The birefringence was found to be very sensitive to the polymersome morphology, as determined by electron microscopy. The deflation of polymersomes into disks was observed, followed by a bending and partial inflation into stomatocytes.


Solar-panel and parasol strategies shape the proteorhodopsin distribution pattern in marine Flavobacteriia.
Proteorhodopsin (PR) is a light-driven proton pump that is found in diverse bacteria and archaea species, and is widespread in marine microbial ecosystems. To date, many studies have suggested the advantage of PR for microorganisms in sunlit environments. The ecophysiological significance of PR is still not fully understood however, including the drivers of PR gene gain, retention, and loss in different marine microbial species. To explore this question we sequenced 21 marine Flavobacteriia genomes of polyphyletic origin, which encompassed both PR-possessing as well as PR-lacking strains. Here, we show that the possession or alternatively the lack of PR genes reflects one of two fundamental adaptive strategies in marine bacteria. Specifically, while PR-possessing bacteria utilize light energy ("solar-panel strategy"), PR-lacking bacteria exclusively possess UV-screening pigment synthesis genes to avoid UV damage and would adapt to microaerobic environment ("parasol strategy"), which also helps explain why PR-possessing bacteria have smaller genomes than those of PR-lacking bacteria. Collectively, our results highlight the different strategies of dealing with light, DNA repair, and oxygen availability that relate to the presence or absence of PR phototrophy.


Extended input space support vector machine.
In some applications, the probability of error of a given classifier is too high for its practical application, but we are allowed to gather more independent test samples from the same class to reduce the probability of error of the final decision. From the point of view of hypothesis testing, the solution is given by the Neyman-Pearson lemma. However, there is no equivalent result to the Neyman-Pearson lemma when the likelihoods are unknown, and we are given a training dataset. In this brief, we explore two alternatives. First, we combine the soft (probabilistic) outputs of a given classifier to produce a consensus labeling for K test samples. In the second approach, we build a new classifier that directly computes the label for K test samples. For this second approach, we need to define an extended input space training set and incorporate the known symmetries in the classifier. This latter approach gives more accurate results, as it only requires an accurate classification boundary, while the former needs an accurate posterior probability estimate for the whole input space. We illustrate our results with well-known databases.


Activation of vasculogenic progenitor cells by ent-16α,17-dihydroxy-kauran-19-oic acid.
Vasculogenic progenitor cells (VPCs) circulate in the blood and have the ability to differentiate into endothelial cells that make up the lining of blood vessels. Therefore, VPC transplantation is a new strategy for the treatment of ischemic diseases. Because priming/preconditioning of VPCs before transplantation enhances their regenerative potential, the present study investigated whether ent-16α,17-dihydroxy-kauran-19-oic acid (DHK) isolated from Siegesbeckia pubescens could stimulate/activate VPCs in vitro. Therefore, the effect of DHK (1-100 µM concentration) on the proliferation, migration, and tube forming of VPCs was examined in various systems, and related signaling pathways were identified. DHK treatment significantly increased the proliferation, migration, and tube formation of VPCs in a dose-dependent manner. Phosphorylation of extracellular signal-regulated kinase (ERK)1/2 and Akt was significantly increased by DHK, but chemical inhibitors against ERK1/2 (U0126) and Akt (LY294002) significantly attenuated DHK-enhanced proliferation, migration, and tube formation of VPCs. Collectively, these results indicated that DHK shows promise as a novel VPC primer/activator.


Use of rifaximin in gastrointestinal and liver diseases.
Rifaximin is a broad spectrum oral antibiotic with antimicrobial activity against Gram-positive and Gram-negative aerobic and anaerobic bacteria. It is poorly absorbed and thus has a highly favorable safety profile. Rifaximin has been shown to be effective in the treatment of traveler's diarrhea, functional bloating and irritable bowel syndrome, small bowel bacterial overgrowth and in the prevention of recurrent overt hepatic encephalopathy. In addition, there is emerging evidence for a possible beneficial effect of rifaximin in the treatment of uncomplicated diverticular disease and in the prevention of recurrent diverticulitis. The use of rifaximin is associated with a low incidence of development, or persistence of spontaneous bacterial mutants. Moreover, the development of important drug resistance among extra-intestinal flora during rifaximin therapy is unlikely because of minimal systemic absorption and limited cross-resistance of rifaximin with other antimicrobials. This review addresses the current and emerging role of rifaximin in the treatment of gastrointestinal and liver disorders.


The adverse effect of negative comments about weight and shape from family and siblings on women at high risk for eating disorders.
Our purpose with this work was to examine the relationship between negative comments about weight, shape, and eating and social adjustment, social support, self-esteem, and perceived childhood abuse and neglect. A retrospective study was conducted with 455 college women with high weight and shape concerns, who participated in an Internet-based eating disorder prevention program. Baseline assessments included: perceived family negative comments about weight, shape, and eating; social adjustment; social support; self-esteem; and childhood abuse and neglect. Participants identified 1 of 7 figures representing their maximum body size before age 18 and parental maximum body size. More than 80% of the sample reported some parental or sibling negative comments about their weight and shape or eating. Parental and sibling negative comments were positively associated with maximum childhood body size, larger reported paternal body size, and minority status. On subscales of emotional abuse and neglect, most participants scored above the median, and nearly one third scored above the 90th percentile. In a multivariate analysis, greater parental negative comments were directly related to higher reported emotional abuse and neglect. Maximum body size was also related to emotional neglect. Parental negative comments were associated with lower reported social support by family and lower self-esteem. In college women with high weight and shape concerns, retrospective reports of negative comments about weight, shape, and eating were associated with higher scores on subscales of emotional abuse and neglect. This study provides additional evidence that family criticism results in long-lasting, negative effects.


Survivin Is a transcriptional target of STAT3 critical to estradiol neuroprotection in global ischemia.
Transient global ischemia causes selective, delayed death of hippocampal CA1 pyramidal neurons in humans and animals. It is well established that estrogens ameliorate neuronal death in animal models of focal and global ischemia. However, the role of signal transducer and activator of transcription-3 (STAT3) and its target genes in estradiol neuroprotection in global ischemia remains unclear. Here we show that a single intracerebral injection of 17β-estradiol to ovariectomized female rats immediately after ischemia rescues CA1 neurons destined to die. Ischemia promotes activation of STAT3 signaling, association of STAT3 with the promoters of target genes, and STAT3-dependent mRNA and protein expression of prosurvival proteins in the selectively vulnerable CA1. In animals subjected to ischemia, acute postischemic estradiol further enhances activation and nuclear translocation of STAT3 and STAT3-dependent transcription of target genes. Importantly, we show that STAT3 is critical to estradiol neuroprotection, as evidenced by the ability of STAT3 inhibitor peptide and STAT3 shRNA delivered directly into the CA1 of living animals to abolish neuroprotection. In addition, we identify survivin, a member of the inhibitor-of-apoptosis family of proteins and known gene target of STAT3, as essential to estradiol neuroprotection, as evidenced by the ability of shRNA to survivin to reverse neuroprotection. These findings indicate that ischemia and estradiol act synergistically to promote activation of STAT3 and STAT3-dependent transcription of survivin in insulted CA1 neurons and identify STAT3 and survivin as potentially important therapeutic targets in an in vivo model of global ischemia.


Representing and decomposing genomic structural variants as balanced integer flows on sequence graphs.
The study of genomic variation has provided key insights into the functional role of mutations. Predominantly, studies have focused on single nucleotide variants (SNV), which are relatively easy to detect and can be described with rich mathematical models. However, it has been observed that genomes are highly plastic, and that whole regions can be moved, removed or duplicated in bulk. These structural variants (SV) have been shown to have significant impact on phenotype, but their study has been held back by the combinatorial complexity of the underlying models. We describe here a general model of structural variation that encompasses both balanced rearrangements and arbitrary copy-number variants (CNV). In this model, we show that the space of possible evolutionary histories that explain the structural differences between any two genomes can be sampled ergodically.


Voluntary HIV counselling and testing among men in rural western Uganda: implications for HIV prevention.
Voluntary HIV counselling and testing (VCT) is one of the key strategies in the prevention and control of HIV/AIDS in Uganda. However, the utilization of VCT services particularly among men is low in Kasese district. We therefore conducted a study to determine the prevalence and factors associated with VCT use among men in Bukonzo West health sub-district, Kasese district. A population-based cross-sectional study employing both quantitative and qualitative techniques of data collection was conducted between January and April 2005. Using cluster sampling, 780 men aged 18 years and above, residing in Bukonzo West health sub-district, were sampled from 38 randomly selected clusters. Data was collected on VCT use and independent variables. Focus group discussions (4) and key informant interviews (10) were also conducted. Binary logistic regression was performed to determine the predictors of VCT use among men. Overall VCT use among men was 23.3% (95% CI 17.2-29.4). Forty six percent (95% CI 40.8-51.2) had pre-test counselling and 25.9% (95%CI 19.9-31.9) had HIV testing. Of those who tested, 96% returned for post-test counselling and received HIV results. VCT use was higher among men aged 35 years and below (OR = 2.69, 95%CI 1.77-4.07), the non-subsistence farmers (OR = 2.37, 95%CI 2.37), the couple testing (OR = 2.37, 95%CI 1.02-8.83) and men with intention to disclose HIV test results to sexual partners (OR = 1.64, 95%CI 1.04-2.60). The major barriers to VCT use among men were poor utilization of VCT services due to poor access, stigma and confidentiality of services. VCT use among men in Bukonzo West, Kasese district was low. In order to increase VCT use among men, the VCT programme needs to address HIV stigma and improve access and confidentiality of VCT services. Among the more promising interventions are the use of routine counselling and testing for HIV of patients seeking health care in health units, home based VCT programmes, and mainstreaming of HIV counselling and testing services in community development programmes.


Reward and punishment act as distinct factors in guiding behavior.
Behavior rests on the experience of reinforcement and punishment. It has been unclear whether reinforcement and punishment act as oppositely valenced components of a single behavioral factor, or whether these two kinds of outcomes play fundamentally distinct behavioral roles. To this end, we varied the magnitude of a reward or a penalty experienced following a choice using monetary tokens. The outcome of each trial was independent of the outcome of the previous trial, which enabled us to isolate and study the effect on behavior of each outcome magnitude in single trials. We found that a reward led to a repetition of the previous choice, whereas a penalty led to an avoidance of the previous choice. Surprisingly, the effects of the reward magnitude and the penalty magnitude revealed a pronounced asymmetry. The choice repetition effect of a reward scaled with the magnitude of the reward. In a marked contrast, the avoidance effect of a penalty was flat, not influenced by the magnitude of the penalty. These effects were mechanistically described using a reinforcement learning model after the model was updated to account for the penalty-based asymmetry. The asymmetry in the effects of the reward magnitude and the punishment magnitude was so striking that it is difficult to conceive that one factor is just a weighted or transformed form of the other factor. Instead, the data suggest that rewards and penalties are fundamentally distinct factors in governing behavior.


Functionalized Graphene Oxide Shields Tooth Dentin from Decalcification.
This in vitro study assessed the efficacy of functionalized graphene oxide (f-GO) nanocomposites on the decalcification of dentin, because dental caries of the root surface is becoming one of the new problems in aged society. Hydroxyapatite plates (HAP) and dentin slices were coated with f-GO nanocomposites by comparing them to silver diamine fluoride as a positive control, then treated with decalcification solutions such as ethylenediaminetetraacetic acid and citrate at 37°C for 24 h. Scanning electron microscopy (SEM) revealed significant protection of the surface morphology of HAP and dentin. On the other hand, a cariogenic Streptococcus mutans growth was inhibited by f-GO nanocomposites. In addition, cytotoxicity of them to epithelial cells was much less than that of povidone-iodine, which is commonly used for oral disinfectant. We synthesized 5 different f-GO nanocomposites such as GO-silver (Ag), GO-Ag-calcium fluoride (CaF2), GO-CaF2, GO-zinc, and GO-tricalcium phosphate (Ca3(PO4)2). They were standardized by evaluating under SEM, transmission electron microscopy (TEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS), thermogravimetry analysis (TGA), and Raman spectra after being synthesized in an aseptic technique. The abilities of GO-Ag, GO-Ag-CaF2, and GO-CaF2 nanocomposites were most preventive for decalcification. In addition, GO-Ag and GO-Ag-CaF2 almost completely inhibited S. mutans growth. However, they did not exhibit cytotoxicity to epithelial cells except at the highest concentration (0.1 w/v%) of GO-Ag and GO-Ag-CaF2. Furthermore, these f-GO nanocomposites exhibited less or no discoloration of dentin, although commonly used silver diamine fluoride causes discoloration of dentin to black. Thus, these f-GO nanocomposites are useful to protect dental caries on the tooth root that becomes a social problem in aged society.


A comparison of three methods of decellularization of pig corneas to reduce immunogenicity.
To investigate whether decellularization using different techniques can reduce immunogenicity of the cornea, and to explore the decellularized cornea as a scaffold for cultured corneal endothelial cells (CECs). Transplantation of decellularized porcine corneas increases graft transparency and survival for longer periods compared with fresh grafts. Six-month-old wild-type pig corneas were cut into 100-200 µm thickness, and then decellularized by three different methods: 1) 0.1% sodium dodecyl sulfate (SDS); 2) hypoxic nitrogen (N2); and 3) hypertonic NaCl. Thickness and transparency were assessed visually. Fresh and decellularized corneas were stained with hematoxylin/eosin (H&E), and for the presence of galactose-α1,3-galactose (Gal) and N-glycolylneuraminic acid (NeuGc, a nonGal antigen). Also, a human IgM/IgG binding assay was performed. Cultured porcine CECs were seeded on the surface of the decellularized cornea and examined after H&E staining. All three methods of decellularization reduced the number of keratocytes in the stromal tissue by >80% while the collagen structure remained preserved. No remaining nuclei stained positive for Gal or NeuGc, and expression of these oligosaccharides on collagen was also greatly decreased compared to expression on fresh corneas. Human IgM/IgG binding to decellularized corneal tissue was considerably reduced compared to fresh corneal tissue. The cultured CECs formed a confluent monolayer on the surface of decellularized tissue. Though incomplete, the significant reduction in the cellular component of the decellularized cornea should be associated with a significantly reduced in vivo immune response compared to fresh corneas.


Tools in a clinical information system supporting clinical trials at a Swiss University Hospital.
Issues concerning inadequate source data of clinical trials rank second in the most common findings by regulatory authorities. The increasing use of electronic clinical information systems by healthcare providers offers an opportunity to facilitate and improve the conduct of clinical trials and the source documentation. We report on a number of tools implemented into the clinical information system of a university hospital to support clinical research. In 2011/2012, a set of tools was developed in the clinical information system of the University Hospital Zurich to support clinical research, including (1) a trial registry for documenting metadata on the clinical trials conducted at the hospital, (2) a patient-trial-assignment-tool to tag patients in the electronic medical charts as participants of specific trials, (3) medical record templates for the documentation of study visits and trial-related procedures, (4) online queries on trials and trial participants, (5) access to the electronic medical records for clinical monitors, (6) an alerting tool to notify of hospital admissions of trial participants, (7) queries to identify potentially eligible patients in the planning phase as trial feasibility checks and during the trial as recruitment support, and (8) order sets to facilitate the complete and accurate performance of study visit procedures. The number of approximately 100 new registrations per year in the voluntary trial registry in the clinical information system now matches the numbers of the existing mandatory trial registry of the hospital. Likewise, the yearly numbers of patients tagged as trial participants as well as the use of the standardized trial record templates increased to 2408 documented trial enrolments and 190 reports generated/month in the year 2013. Accounts for 32 clinical monitors have been established in the first 2 years monitoring a total of 49 trials in 16 clinical departments. A total of 15 months after adding the optional feature of hospital admission alerts of trial participants, 107 running trials have activated this option, including 48 out of 97 studies (49.5%) registered in the year 2013, generating approximately 85 alerts per month. The popularity of the presented tools in the clinical information system illustrates their potential to facilitate the conduct of clinical trials. The tools also allow for enhanced transparency on trials conducted at the hospital. Future studies on monitoring and inspection findings will have to evaluate their impact on quality and safety.


A comparison of rapid enzyme immunoassay tests for the detection of Chlamydia trachomatis cervical infections.
Two rapid enzyme immunoassay test kits were compared with culture for the detection of Chlamydia trachomatis endocervical infections. Endocervical samples for C trachomatis culture and the two enzyme immunoassay tests were evaluated from 502 county health department and Planned Parenthood patients. The prevalence of infection in this population was 12%. Sensitivity and specificity of the Abbott TestPack Chlamydia were 51.7% and 99.5%, respectively, and of the Kodak Surecell Chlamydia were 76.7% and 98.6%, respectively. The positive and negative predictive values for TestPack were 93.9% and 93.8%, and for Surecell were 88.5% and 96.9%, respectively. Additionally, an in vitro investigation was used to evaluate whether typical office staff (physicians, nurses, medical technicians, receptionists, and radiology technicians) were able to perform competently the tests in each kit. Office personnel tested 12 dilutions of a C trachomatis stock sample or negative control sample as unknowns for each kit in the in vitro investigation. There were no differences among office staff in performance when compared for each test kit. Selective use of these enzyme immunoassay tests for high-risk patients in a family practice population that has a high prevalence of patients with C trachomatis infection may be helpful when rapid test results are required and cultures are not feasible. After appropriate training, most physician office personnel were equally able to perform the enzyme immunoassay tests evaluated.


Relationship of the Brief UCSD Performance-based Skills Assessment (UPSA-B) to multiple indicators of functioning in people with schizophrenia and bipolar disorder.
This study assessed the relationship between multiple indicators of 'real-world' functioning and scores on a brief performance-based measure of functional capacity known as the Brief University of California San Diego (UCSD) Performance-based Skills Assessment (UPSA-B) in a sample of 205 patients with either serious bipolar disorder (n = 89) or schizophrenia (n = 116). Participants were administered the UPSA-B and assessed on the following functional domains: (i) independent living status (e.g., residing independently as head of household, living in residential care facility); (ii) informant reports of functioning (e.g., work skills, daily living skills); (iii) educational attainment and estimated premorbid IQ as measured by years of education and Wide Range Achievement Test reading scores, respectively; and (iv) employment. Better scores on the UPSA-B were associated with greater residential independence after controlling for age, diagnosis, and symptoms of psychopathology. Among both bipolar disorder and schizophrenia patients, higher UPSA-B scores were significantly related to better informant reports of functioning in daily living skills and work skills domains. Greater estimated premorbid IQ was associated with higher scores on the UPSA-B for both schizophrenia and bipolar disorder participants. Participants who were employed scored higher on the UPSA-B when controlling for age and diagnosis, but not when controlling for symptoms of psychopathology. These data suggest the UPSA-B may be useful for assessing capacity for functioning in a number of domains in both people diagnosed with schizophrenia and bipolar disorder.


Sterilization by gamma radiation of antibiotic impregnated polymethylmethacrylate and plaster of Paris beads. A pilot study.
Ethylene oxide is currently recommended for sterilization of antibiotic impregnated beads; however this method carries health risks to personnel and is becoming less available. To perform a pilot study of the effect of radiation for sterilization of polymethylmethacrylate (PMMA) and plaster of Paris (POP) beads impregnated with amikacin, enrofloxacin, and ceftiofur. Radiation would effectively sterilize the beads without affecting the efficacy of the antibiotic. Beads of PMMA and POP were prepared in a clean but non-sterile manner with one of the three antibiotics (amikacin, enrofloxacin, ceftiofur) or no antibiotic. Beads were then exposed to radiation for a total dose of 0 kiloGray (kGy), 10 kGy and 25 kGy. Beads were incubated on Mueller-Hinton agar plates seeded with Escherichia coli, Staphylococcus aureus or Pseudomonas aeruginosa for 24 hours or cultured in brain-heart infusion broth for 48 hours. Zones of inhibition were measured on the agar plates and statistics were performed on the diameters of the zones of inhibition using an analysis of variance. There were no differences in the diameters of inhibition for all levels of radiation for all PMMA beads. The same was true with POP beads with the exception of enrofloxacin which had a significantly decreased zone of inhibition with increased levels of radiation, though the clinical significance of this finding was not assessed. Only beads without antibiotics and not exposed to radiation had bacterial growth. Radiation may be an effective method of sterilization for antibiotic impregnated beads.


Urbanization of black South African women may increase risk of low bone mass due to low vitamin D status, low calcium intake, and high bone turnover.
Globally, rural to urban migration is accompanied by changes in dietary patterns and lifestyle that have serious health implications, including development of low bone mass. We hypothesized that serum 25 (OH) vitamin D3 (25[OH]D3) levels will be lower, bone turnover higher, and nutrition inadequate in urban postmenopausal black women, increasing risk for low bone mass. We aimed to assess the prevalence of risk factors for low bone mass in 1261 black women from rural and urban areas in the North West Province of South Africa (Prospective Urban and Rural Epidemiology-South Africa project). Fasting blood samples were taken; and participants were interviewed to complete questionnaires on self-reported diseases, fractures, and dietary intakes. Bone health markers were assessed in a subgroup of 658 women older than 45 years. Specific lifestyle risk factors identified were inactivity, smoking, injectable progestin contraception use, and high alcohol consumption. Dietary risk factors identified were low calcium and high animal protein, phosphorous, and sodium intakes. The 25(OH)D3 and C-terminal telopeptide (CTX) levels were significantly higher in the rural vs the urban women older than 50 years. Parathyroid hormone (PTH) levels increased with age in both groups. The 25(OH)D levels were inversely correlated with CTX and PTH in rural women. In urban women, PTH and CTX were correlated while dietary calcium was inversely correlated with CTX and PTH with 25(OH)D3. The combination of low dietary calcium (<230 mg/d), marginally insufficient 25(OH)D3 status, and raised PTH may result in increased bone resorption. Further research is required to assess bone health and fracture risk in black African women.


An analysis of neurosurgical practice patterns and outcomes for serious to critical traumatic brain injuries in a mature trauma state.
We sought to characterize trends in neurosurgical practice patterns and outcomes for serious to critical traumatic brain injuries from 2003 to 2013 in the mature trauma state of Pennsylvania. All 2003 to 2013 admissions to Pennsylvania's 30 accredited Level I to II trauma centers with serious to critical traumatic brain injuries (head Abbreviated Injury Scale [AIS] score ≥ 3, Glasgow Coma Scale [GCS] score < 13) were extracted from the state registry. Adjusted temporal trend tests controlling for demographic and injury severity covariates assessed the impact of admission year on intervention rates (craniotomy, craniectomy, and intracranial pressure monitor/ventriculostomy [ICP]) and outcome measures for the total population as well as serious (head AIS score ≥ 3; GCS score, 9-12) and critical (head AIS score ≥ 3, GCS score ≤ 8) subgroups. A total of 22,229 patients met inclusion criteria. Admission year was significantly associated with an adjusted increase in craniectomy (adjusted odds ratio [AOR], 1.12 [1.09-1.14]; p < 0.001) and ICP rates (AOR, 1.03 [1.02-1.04]; p < 0.001) and a decrease in craniotomy rate (AOR, 0.96 [0.95-0.97]; p < 0.001). No significant trends in adjusted mortality were found for the total study population (AOR, 1.01 [1.00-1.02]; p = 0.150); however, a significant reduction was found for the serious subgroup (AOR, 0.95 [0.92-0.98]; p = 0.002), and a significant increase was found for the critical subgroup (AOR, 1.02 [1.01-1.03]; p = 0.004). Total study population trends showed a reduction in rates of craniotomy and increase in craniectomy and ICP rates without any change in outcome. Despite significant adaptations in neurosurgical practice patterns from 2003 to 2013, only patients with serious head injuries are experiencing improved survival. Prognostic and epidemiologic study, level III; therapeutic study, level IV.


Non-invasive single-cell biomechanical analysis using live-imaging datasets.
The physiological state of a cell is governed by a multitude of processes and can be described by a combination of mechanical, spatial and temporal properties. Quantifying cell dynamics at multiple scales is essential for comprehensive studies of cellular function, and remains a challenge for traditional end-point assays. We introduce an efficient, non-invasive computational tool that takes time-lapse images as input to automatically detect, segment and analyze unlabeled live cells; the program then outputs kinematic cellular shape and migration parameters, while simultaneously measuring cellular stiffness and viscosity. We demonstrate the capabilities of the program by testing it on human mesenchymal stem cells (huMSCs) induced to differentiate towards the osteoblastic (huOB) lineage, and T-lymphocyte cells (T cells) of naïve and stimulated phenotypes. The program detected relative cellular stiffness differences in huMSCs and huOBs that were comparable to those obtained with studies that utilize atomic force microscopy; it further distinguished naïve from stimulated T cells, based on characteristics necessary to invoke an immune response. In summary, we introduce an integrated tool to decipher spatiotemporal and intracellular dynamics of cells, providing a new and alternative approach for cell characterization.


[Chemical libraries dedicated to protein-protein interactions].
The identification of complete networks of protein-protein interactions (PPI) within a cell has contributed to major breakthroughs in understanding biological pathways, host-pathogen interactions and cancer development. As a consequence, PPI have emerged as a new class of promising therapeutic targets. However, they are still considered as a challenging class of targets for drug discovery programs. Recent successes have allowed the characterization of structural and physicochemical properties of protein-protein interfaces leading to a better understanding of how they can be disrupted with small molecule compounds. In addition, characterization of the profiles of PPI inhibitors has allowed the development of PPI-focused libraries. In this review, we present the current efforts at developing chemical libraries dedicated to these innovative targets.


Statistical analysis of the Indus script using n-grams.
The Indus script is one of the major undeciphered scripts of the ancient world. The small size of the corpus, the absence of bilingual texts, and the lack of definite knowledge of the underlying language has frustrated efforts at decipherment since the discovery of the remains of the Indus civilization. Building on previous statistical approaches, we apply the tools of statistical language processing, specifically n-gram Markov chains, to analyze the syntax of the Indus script. We find that unigrams follow a Zipf-Mandelbrot distribution. Text beginner and ender distributions are unequal, providing internal evidence for syntax. We see clear evidence of strong bigram correlations and extract significant pairs and triplets using a log-likelihood measure of association. Highly frequent pairs and triplets are not always highly significant. The model performance is evaluated using information-theoretic measures and cross-validation. The model can restore doubtfully read texts with an accuracy of about 75%. We find that a quadrigram Markov chain saturates information theoretic measures against a held-out corpus. Our work forms the basis for the development of a stochastic grammar which may be used to explore the syntax of the Indus script in greater detail.


Host immune responses to the intracellular bacteria Brucella: does the bacteria instruct the host to facilitate chronic infection?
Brucella spp. are intracellular gram-negative bacteria that include a number of virulent species that cause chronic infections in a variety of mammalian hosts. Human infections are proportional to the level of disease in domestic animals because humans are infected zoonotically after contact with infected animals or their products. The chronicity of infection results from the ability of some brucellae to survive reactive oxygen intermediate and nitric oxide killing in host phagocytes, following which they activate bacterial genes in response to the acidic phagosome environment, prevent phagolysosomal fusion by remodeling the intracellular compartment, and subsequently replicate intracellularly. The crucial component of immunity that results in survival of the host and thus maintenance of this chronic infective state is interferon-gamma (IFN-gamma). Production of IFN-gamma results from the ability of brucella components, including lipid A, to interact with Toll-like receptors for the production of IL-12 and TNF-alpha, although the regulatory cytokine IL-10 is also produced and decreases control of the infection. Although CD4 and CD8 T cells are clearly involved in the production of IFN-gamma, and CD8 T cells may be cytotoxic, a role for NK cells and cytotoxicity in protective immunity to brucellosis has not been substantiated experimentally. Moreover, antibodies have been shown to have a limited role in passive transfer studies.


Oxidative stress in hypothyroid patients and the role of antioxidant supplementation.
The available data concerning oxidant stress and antioxidant capacity in hypothyroidism are scanty and inconclusive. While some authors suggest that tissues may be protected from oxidant damage because of a hypometabolic state in hypothyroidism, others report increased oxidative stress in hypothyroidism. Selenium acts as a cofactor for the thyroid hormone (TH) deiodinases that activate and then deactivate various THs and their metabolites. Selenium may inhibit thyroid autoimmunity. The study was designed, first, to study the impact of oxidative stress in patients of primary hypothyroidism due to autoimmune thyroiditis, by estimation of serum malondialdehyde (MDA) as a biomarker of oxidative stress. Second, to study the change in MDA level pre- and post-L-thyroxine treatment. Finally, to look into the possible role of selenium supplementation on oxidative stress in autoimmune hypothyroidism. Patients attending endocrine outpatient department (OPD) services of IPGMER and SSKM hospital were considered for the study. Sixty treatment-naive adult patients (age > 18 years) with hypothyroidism were included in the study. The patients were divided into two groups, each comprised thirty patients. One group was treated with L-thyroxine and placebo (Group A). The other group received L-thyroxine replacement along with selenium (100 mcg twice a day) as antioxidant supplementation (Group B). The patients were blinded about selenium and placebo. The study duration for both groups was 6 months. The starting dose of L-thyroxine was 1.6 mcg/kg body weight free thyroxine (FT4), and thyroid-stimulating hormone (TSH) was repeated after 12 weeks. L-thyroxine dose adjustments were done if needed. MDA was assessed at the beginning and at the end of the study, i.e., after 6 months of treatment. The control cohort was composed of thirty healthy adults. Only overt hypothyroidism (OH) cases were included in the study. Normality of data was determined using Anderson-Darling test, Shapiro-Wilk test, and QQ plot. P values were calculated using ANOVA and post hoc Bonferroni tests for normally distributed data. Correlation analysis was carried out using Pearson correlation test. P < 0.05 considered to be statistically significant. After treatment in Group A patients, FT4 showed a significant increment while TSH value decreased. MDA level reduced after treatment, (P < 0.001). After treatment in Group B patients, FT4 showed increment while TSH value decreased (P < 0.05). After treatment, there was a drop in estimated MDA level (P < 0.001). MDA level shows a significant drop in both groups after treatment. In Group B, there is more decline in the MDA percentage but did not reach statistical significance. By performing repeated measure MANOVA, no significant difference was found in the MDA levels between the two groups. MDA reduction when expressed as percentage showed reduction of 39.5% in patients of Group A. Similarly, Group B patients showed a percentage reduction of 45.4%. Oxidative stress compounds hypothyroidism. Hypothyroidism is a state of increased oxidative stress. In this study, biomarker, MDA level is high in treatment-naive primary hypothyroid patients. After treatment with L-thyroxine, the stress marker is reduced to a significant extent. MDA can be used as a useful biomarker to measure and monitor oxidative stress. The role of the addition of antioxidant in the form of selenium remained inconclusive.


Testicular expression of small ubiquitin-related modifier-1 (SUMO-1) supports multiple roles in spermatogenesis: silencing of sex chromosomes in spermatocytes, spermatid microtubule nucleation, and nuclear reshaping.
SUMO-1 is a member of a ubiquitin-related family of proteins that mediates important post-translational effects affecting diverse physiological functions. Whereas SUMO-1 is detected in the testis, little is known about its reproductive role in males. Herein, cell-specific SUMO-1 was localized in freshly isolated, purified male germ cells and somatic cells of mouse and rat testes using Western analysis, high-resolution single-cell bioimaging, and in situ confocal microscopy of seminiferous tubules. During germ cell development, SUMO-1 was observed at low but detectable levels in the cytoplasm of spermatogonia and early spermatocytes. SUMO-1 appeared on gonosomal chromatin during zygotene when chromosome homologues pair and sex chromatin condensation is initiated. Striking SUMO-1 increases in the sex body of early-to-mid-pachytene spermatocytes correlated with timing of additional sex chromosome condensation. Before the completion of the first meiotic division, SUMO-1 disappeared from the sex body when X and Y chromosomal activity resumed. Together, these data indicate that sumoylation may be involved in non-homologous chromosomal synapsis, meiotic sex chromosome inactivation, and XY body formation. During spermiogenesis, SUMO-1 localized in chromocenters of certain round spermatids and perinuclear ring and centrosomes of elongating spermatids, data implicating SUMO-1 in the process of microtubule nucleation and nuclear reshaping. STAT-4, one potential target of sumoylation, was located along the spermatid nuclei, adjacent but not co-localized with SUMO-1. Androgen receptor-positive Leydig, Sertoli, and some peritubular myoepithelial cells express SUMO-1, findings suggesting a role in modulating steroid action. Testicular SUMO-1 expression supports its specific functions in inactivation of sex chromosomes during meiosis, spermatid microtubule nucleation, nuclear reshaping, and gene expression.


Homeodomain proteins Mox1 and Mox2 associate with Pax1 and Pax3 transcription factors.
Mox1 and Mox2 homeobox genes have been shown to be critical in axial skeleton and in limb muscle development respectively. Pax1 and Pax3 gene products are also implicated in these processes. Mox and Pax expression patterns are highly overlapping both spatially and temporally during embryonic development. We show here for the first time that Mox proteins physically interact with Pax1 and Pax3 using the yeast two-hybrid protein interaction assay as well as in vitro biochemical assays. There is a strong preference of Mox1 to associate with Pax1 rather than Pax3 and of Mox2 to associate with Pax3 rather than Pax1. The observed interactions are mediated through the homeodomain of Mox.


The importance of global studies of the genetics of type 2 diabetes.
Genome wide association analyses have revealed large numbers of common variants influencing predisposition to type 2 diabetes and related phenotypes. These studies have predominantly featured European populations, but are now being extended to samples from a wider range of ethnic groups. The transethnic analysis of association data is already providing insights into the genetic, molecular and biological causes of diabetes, and the relevance of such studies will increase as human discovery genetics increasingly moves towards sequencing-based approaches and a focus on low frequency and rare variants.


The adaptive function of masturbation in a promiscuous African ground squirrel.
Studies of animal mating systems increasingly emphasize female multiple mating and cryptic sexual selection, particularly sperm competition. Males under intense sperm competition may manipulate sperm quantity and quality through masturbation, which could waste sperm and decrease fertility. I examined the factors influencing masturbation by male Cape ground squirrels (Xerus inauris) in light of a number of functional hypotheses. Observational data on a marked population of squirrels were collected in east-central Namibia using scan and all-occurrences sampling. Masturbation was far more frequent on days of female oestrus and mostly occurred after copulation. Masturbation rates were higher in dominant males, which copulate more, than in subordinates and increased with number of mates a female accepts. These results suggest that masturbation in this species was not a response to sperm competition nor a sexual outlet by subordinates that did not copulate. Instead masturbation could function as a form of genital grooming. Female Cape ground squirrels mate with up to 10 males in a 3-hr oestrus, and by masturbating after copulation males could reduce the chance of infection. Sexually transmitted infections (STIs) can profoundly affect fertility, and their consequences for mating strategies need to be examined more fully.


A first vascularized skin equivalent as an alternative to animal experimentation.
Tissue-engineered skin equivalents mimic key aspects of the human skin, and can thus be employed as wound coverage for large skin defects or as in vitro test systems as an alternative to animal models. However, current skin equivalents lack a functional vasculature limiting clinical and research applications. This study demonstrates the generation of a vascularized skin equivalent with a perfused vascular network by combining a biological vascularized scaffold (BioVaSc) based on a decellularized segment of a porcine jejunum and a tailored bioreactor system. Briefly, the BioVaSc was seeded with human fibroblasts, keratinocytes, and human microvascular endothelial cells. After 14 days at the air-liquid interface, hematoxylin & eosin and immunohistological staining revealed a specific histological architecture representative of the human dermis and epidermis including a papillary-like architecture at the dermal-epidermal-junction. The formation of the skin barrier was measured non-destructively using impedance spectroscopy. Additionally, endothelial cells lined the walls of the formed vessels that could be perfused with a physiological volume flow. Due to the presence of a complex in-vivo-like vasculature, the here shown skin equivalent has the potential for skin grafting and represents a sophisticated in vitro model for dermatological research.


Modeling Hemolytic-Uremic Syndrome: In-Depth Characterization of Distinct Murine Models Reflecting Different Features of Human Disease.
Diarrhea-positive hemolytic-uremic syndrome (HUS) is a renal disorder that results from infections with Shiga-toxin (Stx)-producing Escherichia coli. The aim of this study was to establish well-defined refined murine models of HUS that can serve as preclinical tools to elucidate molecular mechanisms of disease development. C57BL/6J mice were subjected to different doses of Stx2 purified from an E. coli O157:H7 patient isolate. Animals received 300 ng/kg Stx2 and were sacrificed on day 3 to establish an acute model with fast disease progression. Alternatively, mice received 25 ng/kg Stx2 on days 0, 3, and 6, and were sacrificed on day 7 to establish a subacute model with moderate disease progression. Indicated by a rise in hematocrit, we observed dehydration despite volume substitution in both models, which was less pronounced in mice that underwent the 7-day regime. Compared with sham-treated animals, mice subjected to Stx2 developed profound weight loss, kidney dysfunction (elevation of plasma urea, creatinine, and neutrophil gelatinase-associated lipocalin), kidney injury (tubular injury and loss of endothelial cells), thrombotic microangiopathy (arteriolar microthrombi), and hemolysis (elevation of plasma bilirubin, lactate dehydrogenase, and free hemoglobin). The degree of complement activation (C3c deposition), immune cell invasion (macrophages and T lymphocytes), apoptosis, and proliferation were significantly increased in kidneys of mice subjected to the 7-day but not in kidneys of mice subjected to the 3-day regime. However, glomerular and kidney volume remained mainly unchanged, as assessed by 3D analysis of whole mount kidneys using CD31 staining with light sheet fluorescence microscopy. Gene expression analysis of kidneys revealed a total of only 91 overlapping genes altered in both Stx2 models. In conclusion, we have developed two refined mouse models with different disease progression, both leading to hemolysis, thrombotic microangiopathy, and acute kidney dysfunction and damage as key clinical features of human HUS. While intrarenal changes (apoptosis, proliferation, complement deposition, and immune cell invasion) mainly contribute to the pathophysiology of the subacute model, prerenal pathomechanisms (hypovolemia) play a predominant role in the acute model. Both models allow the further study of the pathomechanisms of most aspects of human HUS and the testing of distinct novel treatment strategies.


Phenotyping Mediterranean Durum Wheat Landraces for Resistance to Zymoseptoria tritici in Tunisia.
Durum wheat landraces have huge potential for the identification of genetic factors valuable for improving resistance to biotic stresses. Tunisia is known as a hot spot for Septoria tritici blotch disease (STB), caused by the fungus Zymoseptoria tritici (Z. tritici). In this context, a collection of 3166 Mediterranean durum wheat landraces were evaluated at the seedling and adult stages for STB resistance in the 2016-2017 cropping season under field conditions in Kodia (Tunisia). Unadapted/susceptible accessions were eliminated to reach the final set of 1059 accessions; this was termed the Med-collection, which comprised accessions from 13 countries and was also screened in the 2018-2019 cropping season. The Med-collection showed high frequency of resistance reactions, among which over 50% showed an immune reaction (HR) at both seedling and adult growth stages. Interestingly, 92% of HR and R accessions maintained their resistance levels across the two years, confirming the highly significant correlation found between seedling- and adult-stage reactions. Plant Height was found to have a negative significant effect on adult-stage resistance, suggesting that either this trait can influence disease severity, or that it can be due to environmental/epidemiological factors. Accessions from Italy showed the highest variability, while those from Portugal, Spain and Tunisia showed the highest levels of resistance at both growth stages, suggesting that the latter accessions may harbor novel QTLs effective for STB resistance.


A mesoderm-inducing factor produced by WEHI-3 murine myelomonocytic leukemia cells is activin A.
The first inductive interaction in amphibian development is mesoderm induction, during which a signal from the vegetal hemisphere of the blastula-staged embryo induces mesoderm from overlying equatorial cells. Recently, a number of 'mesoderm-inducing factors' (MIFs), which may be responsible for this interaction, have been discovered. Examples of these MIFs include members of the fibroblast growth factor family as well as members of the TGF-beta superfamily such as TGF-beta 2. In addition to these purified factors, several new sources of mesoderm-inducing activity have been described. One of the most potent of these is the murine myelomonocytic leukemia cell line WEHI-3. Even at high dilutions, conditioned medium from WEHI-3 cells induces isolated Xenopus animal pole regions to form a variety of mesodermal cell types. In this paper we show by several criteria, including N-terminal amino acid sequencing, Northern blotting and various functional assays, that the WEHI-MIF is activin A. Activins are known to modulate the release of follicle-stimulating hormone from cultured anterior pituitary cells and to cause the differentiation of two erythroleukemia cell lines. Our results, along with recent data from other laboratories, indicate that these molecules may also act in early development in the formation of the mesoderm.


Clinical effectiveness and cost-effectiveness of tailored intensive liaison between primary and secondary care to identify individuals at risk of a first psychotic illness (the LEGs study): a cluster-randomised controlled trial.
General practitioners are usually the first health professionals to be contacted by people with early signs of psychosis. We aimed to assess whether increased liaison between primary and secondary care improves the clinical effectiveness and cost-effectiveness of detection of people with, or at high risk of developing, a first psychotic illness. Our Liaison and Education in General Practices (LEGs) study was a cluster-randomised controlled trial of primary care practices (clusters) in Cambridgeshire and Peterborough, UK. Consenting practices were randomly allocated (1:1) to a 2 year low-intensity intervention (a postal campaign, consisting of biannual guidelines to help identify and refer individuals with early signs of psychosis) or a high-intensity intervention, which additionally included a specialist mental health professional who liaised with every practice and a theory-based educational package. Practices were not masked to group allocation. Practices that did not consent to be randomly assigned comprised a practice-as-usual (PAU) group. The primary outcome was number of referrals of patients at high risk of developing psychosis to the early intervention service per practice site. New referrals were assessed clinically and stratified into those who met criteria for high risk or first-episode psychotic illness (FEP; together: psychosis true positives), and those who did not fulfil such criteria for psychosis (false positives). Referrals from PAU practices were also analysed. We assessed cost-effectiveness with decision analytic modelling in terms of the incremental cost per additional true positive identified. The trial is registered at the ISRCTN registry, number ISRCTN70185866. Between Dec 22, 2009, and Sept 7, 2010, 54 of 104 eligible practices provided consent and between Feb 16, 2010, and Feb 11, 2011, these practices were randomly allocated to interventions (28 to low intensity and 26 to high intensity); the remaining 50 practices comprised the PAU group. Two high-intensity practices were excluded from the analysis. In the 2 year intervention period, high-intensity practices referred more FEP cases than did low-intensity practices (mean 1.25 [SD 1.2] for high intensity vs 0.7 [0.9] for low intensity; incidence rate ratio [IRR] 1.9, 95% CI 1.05-3.4, p=0.04), although the difference was not statistically significant for individuals at high risk of psychosis (0.9 [1.0] vs 0.5 [1.0]; 2.2, 0.9-5.1, p=0.08). For high risk and FEP combined, high-intensity practices referred both more true-positive (2.2 [1.7] vs 1.1 [1.7]; 2.0, 1.1-3.6, p=0.02) and false-positive (2.3 [2.4] vs 0.9 [1.2]; 2.6, 1.3-5.0, p=0.005) cases. Referral patterns did not differ between low-intensity and PAU practices. Total cost per true-positive referral in the 2 year follow-up was £26,785 in high-intensity practices, £27,840 in low-intensity practices, and £30,007 in PAU practices. This intensive intervention to improve liaison between primary and secondary care for people with early signs of psychosis was clinically and cost effective. UK National Institute for Health Research.


Home-like versus conventional institutional settings for birth.
Home-like birth settings have been established in or near conventional labour wards for the care of pregnant women who prefer and require little or no medical intervention during labour and birth. Primary: to assess the effects of care in a home-like birth environment compared to care in a conventional labour ward. Secondary: to determine if the effects of birth settings are influenced by staffing or organizational models or geographical location of the birth centre. We searched the Cochrane Pregnancy and Childbirth Group trials register (18 May 2004) and handsearched eight journals and two published conference proceedings. All randomized or quasi-randomized controlled trials that compared the effects of a home-like institutional birth environment to conventional hospital care. Standard methods of the Cochrane Collaboration Pregnancy and Childbirth Group were used. Two review authors evaluated methodological quality. Double data entry was performed. Results are presented using relative risks and 95% confidence intervals. Six trials involving 8677 women were included. No trials of freestanding birth centres were found. Between 29% and 67% of women allocated to home-like settings were transferred to standard care before or during labour. Allocation to a home-like setting significantly increased the likelihood of: no intrapartum analgesia/anaesthesia (four trials; n = 6703; relative risk (RR) 1.19, 95% confidence interval (CI) 1.01 to 1.40), spontaneous vaginal birth (five trials; n = 8529; RR 1.03, 95% CI 1.01 to 1.06), vaginal/perineal tears (four trials; n = 8415; RR 1.08, 95% CI 1.03 to 1.13), preference for the same setting the next time (one trial; n = 1230; RR 1.81, 95% CI 1.65 to 1.98), satisfaction with intrapartum care (one trial; n = 2844; RR 1.14, 95% CI 1.07 to 1.21), and breastfeeding initiation (two trials; n = 1431; RR 1.05, 95% CI 1.02 to 1.09) and continuation to six to eight weeks (two trials; n = 1431; RR 1.06, 95% CI 1.02 to 1.10). Allocation to a home-like setting decreased the likelihood of episiotomy (five trials; n = 8529; RR 0.85, 95% CI 0.74 to 0.99). There was a trend towards higher perinatal mortality in the home-like setting (five trials; n = 8529; RR 1.83, 95% CI 0.99 to 3.38). No firm conclusions could be drawn regarding the effects of staffing or organizational models. When compared to conventional institutional settings, home-like settings for childbirth are associated with modest benefits, including reduced medical interventions and increased maternal satisfaction. Caregivers and clients should be vigilant for signs of complications.


The civic turn of immigrant integration policies in the Scandinavian welfare states.
This special issue addresses the question of how to understand the civic turn within immigrant integration in the West towards programs and instruments, public discourses and political intentions, which aim to condition, incentivize, and shape through socialization immigrants into 'citizens'. Empirically, it focuses on the less studied Scandinavian cases of Sweden, Norway, and Denmark. In this introduction, we situate the contributions to this special issue within the overall debate on civic integration and convergence. We introduce the three cases, critically discuss the (liberal) convergence thesis and its descriptive and explanatory claims, and explain why studying the Scandinavian welfare states can further our understanding of the nature of the civic turn and its driving forces. Before concluding, we discuss whether civic integration policies actually work.


Ethanol consumption impairs vestibulo-ocular reflex function measured by the video head impulse test and dynamic visual acuity.
Ethanol affects many parts of the nervous system, from the periphery to higher cognitive functions. Due to the established effects of ethanol on vestibular and oculomotor function, we wished to examine its effect on two new tests of the vestibulo-ocular reflex (VOR): the video head impulse test (vHIT) and dynamic visual acuity (DVA). We tested eight healthy subjects with no history of vestibular disease after consumption of standardized drinks of 40% ethanol. We used a repeated measures design to track vestibular function over multiple rounds of ethanol consumption up to a maximum breath alcohol concentration (BrAC) of 1.38 per mil. All tests were normal at baseline. VOR gain measured by vHIT decreased by 25% at the highest BrAC level tested in each subject. Catch-up saccades were negligible at baseline and increased in number and size with increasing ethanol consumption (from 0.13° to 1.43° cumulative amplitude per trial). DVA scores increased by 86% indicating a deterioration of acuity, while static visual acuity (SVA) remained unchanged. Ethanol consumption systematically impaired the VOR evoked by high-acceleration head impulses and led to a functional loss of visual acuity during head movement.


Spatial pattern analysis for water quality in free-surface constructed wetland.
Free-surface constructed wetlands are known as a low-energy green technique to highly decrease a wide range of pollutants in wastewater and stormwater before discharge into natural water. In this study, two spatial analyses, principal factor analysis and hierarchical cluster analysis (HACA), were employed to interpret the effect of wetland on the water quality variables (WQVs) and to classify the wetland into groups with similar characteristics. Eleven WQVs were collected at the 17 sampling stations twice a month for 13 months. All sampling stations were classified by HACA into three clusters, with high, moderate, and low pollution areas. To improve the water quality, the performance of Cluster-III (micropool) is more significant than Cluster-I and Cluster-II. Implications of this study include potential savings of time and cost for long-term data monitoring purposes in the free-constructed wetland.


Miocene burrows of extinct bear dogs: indication of early denning behavior of large Mammalian carnivores.
Partial skeletons of four species of extinct carnivores have been found in their dens excavated in the floodplain of an early Miocene ephemeral braided stream at Agate Fossil Beds National Monument, Nebraska. Bear dogs (Carnivora: Amphicyonidae) were the principal occupants; their den dimensions and body size are similar to those of living wolves and hyenas. Discovery of this predator community extends the record of denning behavior of large mammalian carnivores to the early Miocene, 20 million years ago.


Textures of the soils and rocks at Gusev Crater from Spirit's Microscopic Imager.
The Microscopic Imager on the Spirit rover analyzed the textures of the soil and rocks at Gusev crater on Mars at a resolution of 100 micrometers. Weakly bound agglomerates of dust are present in the soil near the Columbia Memorial Station. Some of the brushed or abraded rock surfaces show igneous textures and evidence for alteration rinds, coatings, and veins consistent with secondary mineralization. The rock textures are consistent with a volcanic origin and subsequent alteration and/or weathering by impact events, wind, and possibly water.


Cancer mortality, state mean elevations, and other selected predictors.
This ecological inquiry compares cancer mortality rates in the U.S. to the predictor of natural background radiation (via land elevation means) along with eight other predictors thought to be associated with cancer mortality. Age-adjusted cancer mortality in 2006 was compared to the predictors of mean land elevation, percent of smokers, educational attainment, percent of population without health insurance, income, obesity, health perception, physical activity, and diet. Among the six predictors considered appropriate for multiple linear regression, three were found to be statistically significant; from strongest to weakest, these three were: smoking, land elevation, and educational attainment. The predictors of smoking and educational attainment have long been considered associated with cancer mortality. The finding that the predictor of land elevation / natural background radiation is inversely related to cancer mortality is another piece of evidence supporting the theory of radiation hormesis. In this study, land elevation / natural background radiation ranked second in predictive strength regarding cancer mortality, behind smoking and ahead of educational attainment. Since this is an ecological inquiry, no causal inferences can be made.


Using a virtual reality system to study balance and walking in a virtual outdoor environment: a pilot study.
Falls and fall-related injuries are a major problem for elderly persons. Most falls occur during walking and turning, and the risk of falling increases when attention is diverted to something besides walking. It is often difficult to standardize methods for testing balance and fall tendency in a clinically relevant setting. We describe the development of a system using a virtual environment (VE) to assess how attention demanding and unexpected events influence a person's capacity to control balance and movement. The hardware in the system consists of a head-mounted display (HMD), a magnetic tracker system, and two SGI computers. The software consists of the image generation of the VE and the management and visualization of motion tracking data. In a preliminary pilot study eight subjects (age 23-80) participated. Each subject walked on a normal floor and was visually presented a familiar outdoor environment in the HMD. They were exposed to different unexpected events, such as a virtual snowfall and tilting of the VE. Disturbances of balance and walking patterns such as changes in speed, stride length and balance reactions like slipping were observed. Two subjects experienced symptoms of cyber sickness with a SSQ score above 25 points. Walking with sensors only did not affect walking time, but in VE the subjects generally walked more slowly. Virtual tilting of the environment had an impact on balance performance during walking. This effect was not observed while the test subjects were walking in a virtual snowfall. The model needs further development but may hold a potential for clinical use.


Asthma control, adiposity, and adipokines among inner-city adolescents.
There is an association between adiposity and asthma prevalence, but the relationship to asthma control is unclear. We sought to understand the relationships among adiposity, sex, and asthma control in inner-city adolescents with asthma. We prospectively followed 368 adolescents with moderate-to-severe asthma (ages 12-20 years) living in 10 urban areas for 1 year. Asthma symptoms and exacerbations were recorded, and pulmonary function and exhaled nitric oxide levels were measured every 6 weeks. Adiposity measures (body mass index [BMI] and dual-energy X-ray absorptiometric scans) were made, and blood was collected for measurement of allergy markers, adiponectin, leptin, TNF-alpha, IL-6, and C-reactive protein levels. More than 60% of female subjects and 50% of male subjects were above the 85th percentile of BMI for age. Higher BMI was associated with more symptom days (R = 0.18, P = .02) and exacerbations (R = 0.18, P = .06) among female subjects only. Adiponectin was inversely related to asthma symptoms (R = -0.18, P < .05) and exacerbations (R = -0.20, P < .05) and positively with FEV(1)/forced vital capacity ratio (R = 0.15, P < .05) in male subjects only independent of body size. There was no relationship between adiposity or adipokines and total IgE levels, blood eosinophil counts, and exhaled nitric oxide levels. Dual-energy X-ray absorptiometry provided little additional value in relating adiposity to asthma outcome in this population of adolescents. Adiposity is associated with poorer asthma control in female subjects. Adiponectin is associated with improved asthma control in male subjects.


Public Daycare Noncompliance with Prescribed Lunch Menus and Dietary Guidelines.
In Brazil, menus for public child daycare centers (PDC) must be planned by a nutritionist in order to meet the infants' nutritional needs and to conform to dietary recommendations. Failure to follow them may jeopardize the infants' health and growth. Therefore, the objectives of this study were to investigate the compliance of actually served lunch menus with the prescribed menus, according to age (7- to 12-month-olds and 13- to 24-month-olds) and whether prescribed and served menus followed the recommended dietary guidelines. Five PDCs were randomly selected for observation, out of 14 existing in the city of Concordia, Brazil. Data collection was carried out during 6 consecutive weeks in September (fall/winter menus) and October (spring/summer menus) in order to cover the menus representative of the entire year. Of 60 lunches recorded, only 20% of them matched the prescribed ones entirely; none of the lunches served to infants aged 7-12 months did so. Fourteen food items were prescribed 157 times throughout the year. Omission (number of times on menu but not served) was the most frequent form of noncompliance in the younger group: 62.4% (98/157). Foods more frequently omitted by the cooks were beef, pork, chicken, and lentils. Compliance with dietary guidelines was higher in the prescribed menus and in those actually served to the 13- to 24-month-old age group. Infants in the 7- to 12-month-old group may be more vulnerable to nutritional inadequacies because menus served to them were less compliant with the prescribed menus. Dietetics professionals should improve the variety of foods on the menus as well as supervise their execution.


Stigma and beliefs of efficacy towards traditional Chinese medicine and Western psychiatric treatment among Chinese-Americans.
In this study we examined community attitudes of efficacy and shame to investigate the factors that may underlie mental health service underutilization among Chinese Americans. We administered an experimental vignette to assess beliefs of efficacy and shame toward using traditional Chinese medicine (TCM) as opposed to Western health services in a sample of 90 Chinese Americans obtained through a national telephone survey. Contrary to predictions, beliefs of efficacy for treating mental disorders were found to be greater for Western psychiatric services when compared with TCM. As predicted however, Chinese Americans perceived greater community attitudes of shame when accessing Western psychiatric services as opposed to TCM to treat a mental disorder. Furthermore, these differences in community attitudes of shame only occurred among respondents who received a psychiatric condition vignette, but not with respondents who were administered a vignette of a physical disorder. These findings suggest that perceived shame may play an important role in help seeking and that integration of TCM with Western psychiatric care may have implications for reducing shame for Chinese Americans.


Preoperative serum retinol-binding protein 4 is associated with the prognosis of patients with hepatocellular carcinoma after curative resection.
Metabolic syndrome and insulin resistance have been linked to increased risk of occurrence and mortality of hepatocellular carcinoma (HCC). Recently, retinol-binding protein 4 (RBP4) was clarified as a specific serological marker of insulin resistance. The aim of this study was to determine whether serum RBP4 could be used as a potential marker for predicting prognosis in patients with HCC after curative resection. Western immunoblotting and Enzyme-linked immunosorbent assay were used to measure the RBP4 expression in cell lines, supernatant, and serum. Serum RBP4 levels were compared with clinicopathological features and outcomes of patients with HCC. Furthermore, we investigated the impact of serum RBP4 level, serum C-peptide level, and HOMA-IR on overall survival (OS) and disease-free survival (DFS) of patients with HCC in the training cohort (156 patients with HCC), and then were validated in the validation cohort (105 patients with HCC). RBP4 protein overexpressed in HCC cell lines compared with normal liver cell line (P < 0.001) and correlated with metastatic potential. Serum RBP4 levels were associated with OS [hazard ratio (HR) = 2.208, P < 0.001] and DFS (HR = 1.878, P = 0.029) of patients with HCC. By multivariate analysis, the serum RBP4 level was identified as an independent factor for OS (HR = 2.170, P = 0.004) and DFS (HR = 1.769, P = 0.037) of patients with HCC. The prognostic value of serum RBP4 level was confirmed in the validation cohort. The serum RBP4 level is potential to be a useful prognostic factor for HCC after curative resection.


A multicountry health partnership programme to establish sustainable trauma training in east, central, and southern African countries using a cascading trauma management course model.
Injury accounts for 267 000 deaths annually in the nine College of Surgeons of East, Central, and Southern Africa (COSECSA-ASESA) countries, and the introduction of a sustainable standardised trauma training programme across all cadres is essential. We have delivered a primary trauma care (PTC) programme that encompasses both a "provider" and "training the trainers" course using a "cascading training model" across nine COSECSA countries. The first "primary course" in each country is delivered by a team of UK instructors, followed by "cascading courses" to more rural regions led by newly qualified local instructors, with mentorship provided by UK instructors. This study examines the programme's effectiveness in terms of knowledge, clinical confidence, and cost-effectiveness. We collected pre-training and post-training data from 1030 candidates (119 clinical officers, 540 doctors, 260 nurses, and 111 medical students) trained over 28 courses (nine primary and 19 cascading courses) between Dec 5, 2012, and Dec 19, 2013. Knowledge was assessed with a validated PTC multiple choice questionnaire and clinical confidence ratings of eight trauma scenarios, measured against covariants of sex, age, clinical experience, job roles, country, and health institution's workload. Post-training, a significant improvement was noted across all cadres in knowledge (19% [95% CI 18·0-19·5]; p<0·05) and clinical confidence (22% [20·3-22·3]; p<0·05). Non-doctors showed a greater improvement in knowledge (22% vs 16%; p<0·05) and confidence (24% vs 20%; p<0·05) than doctors. Candidates attending cascading courses also showed larger improvements in knowledge (21% vs 15%; p<0·002) and clinical confidence (23% vs 19%; p<0·002) than their primary course counterparts. Multivariate regression analysis showed that attending cascading courses (Coef=4·83, p<0·05), being a nurse (Coef=3·89, p=0·007) or a clinical officer (Coef=4·11, p=0·015), and attending a course in Kenya (Coef=9·55, p<0·002) or Tanzania (Coef=9·40, p<0·002) were strong predictors to improvement in multiple choice questionnaire performance. However, improvement in clinical confidence was affected by the job-role of the clinical officer (Coef=6·49, p=0·002) and attending a course in Kenya (Coef=16·12, p<0·02) or Tanzania (Coef=7·01, p<0·05). Cascading courses were on average £2000 less expensive than primary care courses. To the best of our knowledge, this is the largest series in the literature on multicountry trauma management training in sub-Saharan Africa. Our study supports the concept of cascading courses as an educationally and cost-effective method in delivering vital trauma training in low-resource settings led by local clinicians. Health Partnership Scheme through the UK Department for International Development (DFID).


Once-Daily Topical Dapsone Gel, 7.5%: Effective for Acne Vulgaris Regardless of Baseline Lesion Count, With Superior Efficacy in Females
Introduction: Acne vulgaris is more common in females than males and is challenging to treat. A post hoc analysis of 2 clinical trials evaluated the effect of dapsone gel, 7.5% based on sex and baseline acne lesion count. Methods: Two identically designed, randomized, double-blind, vehicle-controlled, multicenter, 12-week, phase 3 trials enrolled patients aged ≥12 years with facial acne and 20 to 50 inflammatory and 30 to 100 comedonal lesions. Patients applied dapsone gel, 7.5% or vehicle topically once daily for 12 weeks. Baseline to week 12 reductions were evaluated for total, inflammatory, and comedonal lesions in the pooled dapsone population by sex and total baseline acne lesion count (low: 50–74, medium: 75–99, and high: ≥100). Results: The analysis included 2160 patients (56% female, 44% male). Males and females had similar average baseline total, inflammatory, and comedonal lesion counts. Low, medium, and high subgroups experienced efficacy with dapsone gel, 7.5%. Females in each subgroup experienced superior efficacy to males. In females, total lesion counts in the low, medium, and high subgroups decreased by 56.07%, 50.22%, and 47.63%, respectively, compared with 47.95%, 42.30%, and 34.68% in males (P<0.001 for each male-female comparison). Females' respective inflammatory lesion count percentage reductions were 60.96%, 57.91%, and 55.83%, versus 52.75% (P<0.001), 46.85% (P<0.001), and 44.70% (P=0.008) in males. Females' respective comedonal lesion count percentage reductions were 52.96%, 45.40%, and 44.22%, versus 44.67% (P<0.001), 39.38% (P=0.030), and 29.89% (P=0.001) in males. The TEAE rate was low for the overall population (18.3%) and similar for females (19.0%) and males (17.4%). Males and females had similarly favorable dermal tolerability. Conclusion: Once-daily dapsone gel, 7.5% was efficacious for acne regardless of baseline total lesion count, with superior efficacy in females and similar tolerability in males and females. Registration identifier: Clinicaltrials.gov: NCT01974141 and NCT01974323