Regulation of mouse follicle development by follicle-stimulating hormone in a three-dimensional in vitro culture system is dependent on follicle stage and dose.
The developmental requirements of ovarian follicles are dependent on the maturation stage of the follicle; in particular, elegant studies with genetic models have indicated that FSH is required for antral, but not preantral, follicle growth and maturation. To elucidate further the role of FSH and other regulatory molecules in preantral follicle development, in vitro culture systems are needed. We employed a biomaterials-based approach to follicle culture, in which follicles were encapsulated within matrices that were tailored to the specific developmental needs of the follicle. This three-dimensional system was used to examine the impact of increasing doses of FSH on follicle development for two-layered secondary (100-130 microm; two layers of granulosa cells surrounding the oocyte) and multilayered secondary (150-180 microm, several layers of granulosa cells surrounding the oocyte) follicles isolated from mice. Two-layered secondary follicles were FSH responsive when cultured in alginate-collagen I matrices, exhibiting FSH dose-dependent increases in follicle growth, lactate production, and steroid secretion. Multilayered secondary follicles were FSH dependent, with follicle survival, growth, steroid secretion, metabolism, and oocyte maturation all regulated by FSH. However, doses greater than 25 mIU/ml of FSH negatively impacted multilayered secondary follicle development (reduced follicle survival). The present results indicate that the hormonal and environmental needs of the follicular complex change during the maturation process. The culture system can be adapted to each stage of development, which will be especially critical for translation to human follicles that have a longer developmental period.


Narrow Spectrum Kinase Inhibitors Demonstrate Promise for the Treatment of Dry Eye Disease and Other Ocular Inflammatory Disorders.
The purpose of this study is to determine the potential of narrow spectrum kinase inhibitors (NSKIs) to treat inflammatory eye disorders. Human conjunctival epithelial (HCE) cells were retrieved from subjects via impression cytology. Real-time quantitative PCR (qPCR) was performed on HCE cells to determine gene expression of NSKI kinase targets and proinflammatory cytokines in dry eye disease (DED) patients versus healthy controls. qPCR also assessed p38α expression in hyperosmolar-treated Chang conjunctival epithelial cells. Interaction of NSKI TOP1362 with the kinases was evaluated in ATP-dependent Z-LYTE and competition binding assays. Anti-inflammatory activity was assessed in human peripheral blood mononuclear cells and primary macrophages. In an endotoxin-induced uveitis (EIU) study, lipopolysaccharide (LPS) was administered intravitreally to Lewis rats. TOP1362, dexamethasone, or vehicle was administered topically, and inflammatory cytokine levels were measured 6 hours after LPS injection. HCE cells from DED patients showed significantly increased expression of p38α, spleen tyrosine kinase (Syk), Src, lymphocyte-specific protein tyrosine kinase (Lck), interleukin one beta (IL-1β), interleukin eight (IL-8), monocyte chemotactic protein-1 (MCP-1), and matrix metalloproteinase-9 (MMP-9). TOP1362 strongly inhibited the kinase targets p38α, Syk, Src, and Lck, blocked the rise in p38α expression in hyperosmolar Chang cells, and potently reduced inflammatory cytokine release in cellular models of innate and adaptive immunities. In the EIU model, TOP1362 dose-dependently attenuated the LPS-induced rise in inflammatory cell infiltration and ocular cytokine levels with efficacy comparable to that of dexamethasone. TOP1362 is a potent inhibitor of kinases upregulated in DED and markedly attenuates proinflammatory cytokine release in vitro and in vivo, highlighting the therapeutic potential of NSKIs for treating ocular inflammation, such as that observed in DED.


Multi-nutrient vs. nitrogen-only effects on carbon sequestration in grassland soils.
Human activities have greatly increased the availability of biologically active forms of nutrients [e.g., nitrogen (N), phosphorous (P), potassium (K), magnesium (Mg)] in many soil ecosystems worldwide. Multi-nutrient fertilization strongly increases plant productivity but may also alter the storage of carbon (C) in soil, which represents the largest terrestrial pool of organic C. Despite this issue is important from a global change perspective, key questions remain on how the single addition of N or the combination of N with other nutrients might affect C sequestration in human-managed soils. Here, we use a 19-year old nutrient addition experiment on a permanent grassland to test for nutrient-induced effects on soil C sequestration. We show that combined NPKMg additions to permanent grassland have 'constrained' soil C sequestration to levels similar to unfertilized plots whereas the single addition of N significantly enhanced soil C stocks (N-only fertilized soils store, on average, 11 t C ha(-1) more than unfertilized soils). These results were consistent across grazing and liming treatments suggesting that whilst multi-nutrient additions increase plant productivity, soil C sequestration is increased by N-only additions. The positive N-only effect on soil C content was not related to changes in plant species diversity or to the functional composition of the plant community. N-only fertilized grasslands show, however, increases in total root mass and the accumulation of organic matter detritus in topsoils. Finally, soils receiving any N addition (N only or N in combination with other nutrients) were associated with high N losses. Overall, our results demonstrate that nutrient fertilization remains an important global change driver of ecosystem functioning, which can strongly affect the long-term sustainability of grassland soil ecosystems (e.g., soils ability to deliver multiple ecosystem services).


Laser-driven resonance of dye-doped oil-coated microbubbles: A theoretical and numerical study.
Microbubbles are used to enhance the contrast in ultrasound imaging. When coated with an optically absorbing material, these bubbles can also provide contrast in photoacoustic imaging. This multimodal aspect is of pronounced interest to the field of medical imaging. The aim of this paper is to provide a theoretical framework to describe the physical phenomena underlying the photoacoustic response. This article presents a model for a spherical gas microbubble suspended in an aqueous environment and coated with an oil layer containing an optically absorbing dye. The model includes heat transfer between the gas core and the surrounding liquids. This framework is suitable for the investigation of both continuous wave and pulsed laser excitation. This work utilizes a combination of finite difference simulations and numerical integration to determine the dependancy on the physical properties, including composition and thickness of the oil layer on the microbubble response. A normalization scheme for a linearized version of the model was derived to facilitate comparison with experimental measurements. The results show that viscosity and thickness of the oil layer determine whether or not microbubble resonance can be excited. This work also examines the use of non-sinusoidal excitation to promote harmonic imaging techniques to further improve the imaging sensitivity.


State-of-the-art assessment allows for improved vestibular evoked myogenic potential test-retest reliability.
The goal of the present study was to evaluate the test-retest reliability values of myogenic responses using the latest guidelines for vestibular assessment. Twenty-two otologically and neurologically normal adults were assessed twice, on two different days. The analyses were carried out using interclass correlations. The results showed that the latest recommendations for vestibular assessment lead to test-retest reliability values that are as high, or greater, than those reported in previous studies. The results suggest that state-of-the-art testing, using the latest recommendations as well as electromyography control, improves reliability values of myogenic responses, more specifically for the cervical vestibular evoked myogenic potentials. The impact of small differences in experimental procedures on the reliability values of myogenic responses is also addressed.


HiChIP: a high-throughput pipeline for integrative analysis of ChIP-Seq data.
Chromatin immunoprecipitation (ChIP) followed by next-generation sequencing (ChIP-Seq) has been widely used to identify genomic loci of transcription factor (TF) binding and histone modifications. ChIP-Seq data analysis involves multiple steps from read mapping and peak calling to data integration and interpretation. It remains challenging and time-consuming to process large amounts of ChIP-Seq data derived from different antibodies or experimental designs using the same approach. To address this challenge, there is a need for a comprehensive analysis pipeline with flexible settings to accelerate the utilization of this powerful technology in epigenetics research. We have developed a highly integrative pipeline, termed HiChIP for systematic analysis of ChIP-Seq data. HiChIP incorporates several open source software packages selected based on internal assessments and published comparisons. It also includes a set of tools developed in-house. This workflow enables the analysis of both paired-end and single-end ChIP-Seq reads, with or without replicates for the characterization and annotation of both punctate and diffuse binding sites. The main functionality of HiChIP includes: (a) read quality checking; (b) read mapping and filtering; (c) peak calling and peak consistency analysis; and (d) result visualization. In addition, this pipeline contains modules for generating binding profiles over selected genomic features, de novo motif finding from transcription factor (TF) binding sites and functional annotation of peak associated genes. HiChIP is a comprehensive analysis pipeline that can be configured to analyze ChIP-Seq data derived from varying antibodies and experiment designs. Using public ChIP-Seq data we demonstrate that HiChIP is a fast and reliable pipeline for processing large amounts of ChIP-Seq data.


[A case of hypertrophic cranial pachymeningitis developed skull lesion].
We report a case of hypertrophic cranial pachymeningitis (HCP) developed skull lesion. A 70-year-old male presented with the symptom of left hemiconvulsion. MRI revealed that the enhanced intraosseous mass infiltrated into the the dura and brain parenchyma under the parasagittal region of the right parietal bone. Histological examination revealed chronic inflammation with lymphoplasmacytic infiltrate and fibrosis of both intraosseous mass and dural invasive lesion. Steroid therapy resulted in improvement of clinical symptoms and enhanced lesion of MRI. Three years later, the patient presented with generalized convulsion and weakness of right upper and lower limbs. MRI revealed dural thickening with gadolinium enhancement in the bilateral parasagittal region and falx. Angiography showed occlusion of the superior sagittal sinus. The cause of relapsing symptoms in this patient may have been related to the occlusion of the superior sagittal sinus, due to HCP. We considered that the incipient intraosseous mass resulted from a response of the marrow by destructive progression of chronic inflammation passed through the fracture crack or the cavity of arachnoid granulation.


The hypothalamic neuropeptide FF network is impaired in hypertensive patients.
The human hypothalamus contains the neuropeptide FF (NPFF) neurochemical network. Animal experiments demonstrated that NPFF is implicated in the central cardiovascular regulation. We therefore studied expression of this peptide in the hypothalamus of individuals who suffered from essential hypertension (n = 8) and died suddenly due to acute myocardial infarction (AMI), and compared to that of healthy individuals (controls) (n = 6) who died abruptly due to mechanical trauma of the chest. The frozen right part of the hypothalamus was cut coronally into serial sections of 20 μm thickness, and each tenth section was stained immunohistochemically using antibody against NPFF. The central section through each hypothalamic nucleus was characterized by the highest intensity of NPFF immunostaining and thus was chosen for quantitative densitometry. In hypertensive patients, the area occupied by NPFF immunostained neuronal elements in the central sections through the suprachiasmatic nucleus (SCh), paraventricular hypothalamic nucleus (Pa), bed nucleus of the stria terminalis (BST), perinuclear zone (PNZ) of the supraoptic nucleus (SON), dorso- (DMH), ventromedial (VMH) nuclei, and perifornical nucleus (PeF) was dramatically decreased compared to controls, ranging about six times less in the VMH to 15 times less in the central part of the BST (BSTC). The NPFF innervation of both nonstained neuronal profiles and microvasculature was extremely poor in hypertensive patients compared to control. The decreased NPFF expression in the hypothalamus of hypertensive patients might be a cause of impairment of its interaction with other neurochemical systems, and thereby might be involved in the pathogenesis of the disease.


Time-kill kinetics of cadazolid and comparator antibacterial agents against different ribotypes of Clostridium difficile.
Clostridium difficile infection (CDI) is an increasing cause of nosocomial diarrhoea worldwide, which has been partly attributed to the emergence of hypervirulent strains including C. difficile BI/NAP1/ribotype 027 and BK/NAP7/ribotype 078. Cadazolid is a new antibiotic currently in late-stage clinical studies for the treatment of CDI. The present study evaluated the in vitro bactericidal effect of cadazolid and comparator antibiotics against four C. difficile strains. The data demonstrate the potent and bactericidal activity of cadazolid against different ribotypes of C. difficile. MICs for test antibiotics were determined in brain- heart infusion-supplemented broth (BHIS) containing 5 g l-1 yeast extract and 0.025 % (w/v) l-cysteine. Time-kill kinetics to investigate the rate of killing of each antibiotic at sub- and supra-MIC concentrations were performed at concentrations of 0.5, 1, 2, 4, 8 or 16× the MIC of cadazolid, vancomycin and fidaxomicin at intervals over a 48 h period.Results/key findings. Cadazolid-mediated killing of C. difficile was faster and occurred at lower concentrations than observed for vancomycin, while potency and killing was largely comparable to those observed for fidaxomicin. Notably, cadazolid also displayed a potent bactericidal effect against fluoroquinolone-resistant hypervirulent ribotype 027 and 078 strains. C. difficile spore formation was largely inhibited by all three antibiotics at concentrations >1× MIC; however, none were able to eliminate spores effectively, which were present at the start of the experiment. The data presented here demonstrate the potent in vitro bactericidal activity of cadazolid against different ribotypes of C. difficile, although on a limited set of strains.


Spinal ependymomas. Part 1: Intramedullary ependymomas.
OBJECT Ependymomas represent the most common intramedullary tumor in adults. Despite their usually well-defined dissection plane, surgical morbidity has been documented to be considerably higher compared with other intramedullary entities. This study presents an analysis of risk factors for surgical morbidity and data on long-term results for intramedullary ependymomas. METHODS Among 1447 patients with tumors of the spinal canal treated between 1980 and 2014, 309 patients presented with intramedullary tumors. One hundred patients with intramedullary ependymomas underwent 102 operations. Mean age was 44 ± 15 years (range 8-74 years). Patients were followed by outpatient visits and questionnaires, with a mean follow-up of 77 ± 91 months. Short-term results were determined for individual symptoms and the McCormick Scale, whereas tumor recurrence rates were calculated with Kaplan-Meier statistics. RESULTS Compared with cervical ependymomas, those of the thoracic spine were associated with more severe motor deficits and gait problems at presentation. A total of 86.3% of patients with intramedullary ependymomas underwent gross-total resection (GTR). A low preoperative McCormick grade and first surgery were the strongest predictors for a GTR. Postoperatively, 67.6% of patients demonstrated a worse neurological state at discharge from the hospital. This deterioration was transient for 40.1% of the patients and permanent for 27.5%. In the long term, the McCormick grade remained unchanged from the preoperative grade in 74.5% of patients, while it was improved in 5.9% of patients and increased after surgery in 19.6% of patients. According to a multivariate analysis, the risk of permanent morbidity increased with a thoracic level of the ependymoma, advanced age, a long clinical history, presence of a tumor hemorrhage, and surgery on a recurrent tumor. In the long term, tumor recurrence rates correlated significantly with the amount of resection (4.2% and 18.5% in 20 years after GTR and partial resections, respectively). Postoperative neuropathic pain syndromes affected 37.0% of patients, whereas 4% demonstrated a postoperative myelopathy related to cord tethering at the level of surgery. CONCLUSIONS Intramedullary ependymomas are tumors best treated surgically. A complete resection indicates cure for the overwhelming majority of these patients. Surgery should be performed early by neurosurgeons who deal with these lesions on a regular basis to achieve high GTR rates. Permanent surgical morbidity varies most according to tumor location and patient age.


IL-10 provides cardioprotection in diabetic myocardial infarction via upregulation of Heme clearance pathways.
Diabetes is a risk factor for myocardial infarction, and outcomes after myocardial infarction are worse among diabetics compared with nondiabetics. Diabetes is associated with impaired Heme clearance. Here, we determined whether heme toxicity and impaired heme clearance contribute to diabetic myocardial infarction injury and assessed IL-10 as a therapeutic agent for diabetic myocardial infarction. Plasma-free hemoglobin was significantly elevated in diabetic mice compared with nondiabetic mice after myocardial infarction. Infarct size had strong correlation to the level of plasma-free hemoglobin. Hemoglobin and reactive iron deposition within the infarct zone were also demonstrated in diabetic MI. IL-10 significantly reduced infarct size and improved cardiac function in diabetic mice. Moreover, IL-10 improved capillary density, reduced apoptosis, and decreased inflammation in the border zone of the infarcted hearts, findings that were partially inhibited by Tin protoporphyrin (a heme oxygenase-1 inhibitor). IL-10 upregulated CD163, the hemoglobin:haptoglobin scavenger receptor, and heme oxygenase-1 in THP-1-derived and primary human CD14+ macrophages. IL-10 significantly protected against ischemic injury when HL-1 cardiomyocytes were cotreated with hemoglobin. Together, our findings indicate that IL-10 is cardioprotective in diabetic myocardial infarction via upregulation of heme clearance pathways. These findings implicate heme clearance as a potentially novel therapeutic direction for diabetic myocardial infarction.


The illusion of righteousness: corporate social responsibility practices of the alcohol industry.
Corporate social responsibility (CSR) has become an integral element of how the alcohol industry promotes itself. The existing analyses of CSR in the alcohol industry point to the misleading nature of these CSR practices. Yet, research has been relatively sparse on how the alcohol industry advances CSR in an attempt to facilitate underlying business interests, and in what ways the ongoing display of industry CSR impacts public health. This paper aims to investigate the alcohol industry's recent CSR engagements and explain how CSR forms part of the industry's wider political and corporate strategies. Our study used qualitative methods to collect and analyse data. We searched for materials pertaining to CSR activities from websites of three transnational alcohol corporations, social media platforms, media reports and other sources. Relevant documents were thematically analysed with an iterative approach. Our analysis identified three CSR tactics employed by the alcohol companies which are closely tied in with the industry's underlying corporate intents. First, the alcohol manufacturers employ CSR as a means to frame issues, define problems and guide policy debates. In doing this, the alcohol companies are able to deflect and shift the blame from those who manufacture and promote alcoholic products to those who consume them. Second, the alcohol corporations promote CSR initiatives on voluntary regulation in order to delay and offset alcohol control legislation. Third, the alcohol corporations undertake philanthropic sponsorships as a means of indirect brand marketing as well as gaining preferential access to emerging alcohol markets. The increasing penetration and involvement of the alcohol industry into CSR highlights the urgent needs for public health counter actions. Implementation of any alcohol control measures should include banning or restricting the publicity efforts of the industry's CSR and informing the public of the alcohol industry's notion of social responsibility. More significantly, an internationally binding instrument should be called for to enable countries to differentiate between genuine concerns and spurious altruism, and in doing so, resist the industry's attempt to erode alcohol control.


Reduced contact-inhibition and substratum adhesion in epithelial cells expressing GlcNAc-transferase V.
Malignant transformation of fibroblast and epithelial cells is accompanied by increased beta 1-6 N-acetylglucosaminyltransferase V (GlcNAc-TV) activity, a Golgi N-linked oligosaccharide processing enzyme. Herein, we report that expression of GlcNAc-TV in Mv1Lu cells, an immortalized lung epithelial cell line results in loss of contact-inhibition of cell growth, an effect that was blocked by swainsonine, an inhibitor of Golgi processing enzyme alpha-mannosidase II. In serum-deprived and high density monolayer cultures, the GlcNAc-TV transfectants formed foci, maintained microfilaments characteristic of proliferating cells, and also experienced accelerated cell death by apoptosis. Injection of the GlcNAc-TV transfectants into nude mice produced a 50% incidence of benign tumors, and progressively growing tumors in 2:12 mice with a latency of 6 mo, while no growth was observed in mice injected with control cells. In short term adhesion assays, the GlcNAc-TV expressing cells were less adhesive on surfaces coated with fibronectin and collagen type IV, but no changes were observed in levels of cell surface alpha 5 beta 1 or alpha v beta 3 integrins. The larger apparent molecular weights of the LAMP-2 glycoprotein and integrin glycoproteins alpha 5, alpha v and beta 1 in the transfected cells indicates that their oligosaccharide chains are substrates for GlcNAc-TV. The results suggest that beta 1-6GlcNAc branching of N-linked oligosaccharides contributes directly to relaxed growth controls and reduce substratum adhesion in premalignant epithelial cells.


Loss of zolpidem efficacy in the hippocampus of mice with the GABAA receptor gamma2 F77I point mutation.
Zolpidem is a hypnotic benzodiazepine site agonist with some gamma-aminobutyric acid (GABA)(A) receptor subtype selectivity. Here, we have tested the effects of zolpidem on the hippocampus of gamma2 subunit (gamma2F77I) point mutant mice. Analysis of forebrain GABA(A) receptor expression with immunocytochemistry, quantitative [(3)H]muscimol and [(35)S] t-butylbicyclophosphorothionate (TBPS) autoradiography, membrane binding with [(3)H]flunitrazepam and [(3)H]muscimol, and comparison of miniature inhibitory postsynaptic current (mIPSC) parameters did not reveal any differences between homozygous gamma2I77/I77 and gamma2F77/F77 mice. However, quantitative immunoblot analysis of gamma2I77/I77 hippocampi showed some increased levels of gamma2, alpha1, alpha4 and delta subunits, suggesting that differences between strains may exist in unassembled subunit levels, but not in assembled receptors. Zolpidem (1 microm) enhanced the decay of mIPSCs in CA1 pyramidal cells of control (C57BL/6J, gamma2F77/F77) mice by approximately 60%, and peak amplitude by approximately 20% at 33-34 degrees C in vitro. The actions of zolpidem (100 nm or 1 microm) were substantially reduced in gamma2I77/I77 mice, although residual effects included a 9% increase in decay and 5% decrease in peak amplitude. Similar results were observed in CA1 stratum oriens/alveus interneurons. At network level, the effect of zolpidem (10 microm) on carbachol-induced oscillations in the CA3 area of gamma2I77/I77 mice was significantly different compared with controls. Thus, the gamma2F77I point mutation virtually abolished the actions of zolpidem on GABA(A) receptors in the hippocampus. However, some residual effects of zolpidem may involve receptors that do not contain the gamma2 subunit.


Specific dysphoric symptoms are predicted by early maladaptive schemas.
Early maladaptive schemas (EMSs) are cognitive patterns resulting from unmet core emotional needs in childhood that have been linked to the development of psychopathology. As depression is a multifaceted phenomenon, we hypothesized that specific dysphoric symptoms would be predicted by different EMSs. Four hundred and fifty-six participants completed a measure of EMSs (Young Schema Questionnaire) and reported on the severity of the symptoms of criterion A for major depression in DSM-IV during the occurrence of a dysphoric episode in the previous 12 months. A series of stepwise multiple regression analyses were performed to investigate the predictive power of the EMSs for the severity of each specific depressive symptom. When controlling for gender and current levels of depression, specific symptoms were predicted by different EMSs: sadness by Negativity/Pessimism; anhedonia by Failure; self-harm by Emotional Deprivation and Vulnerability to Harm or Illness; worthlessness by Failure and Negativity/Pessimism; psychomotor retardation/restlessness by Vulnerability to Harm or Illness and Entitlement/Grandiosity; and poor concentration by Insufficient Self-Control/Self-Discipline. The more physical symptoms of fatigue, insomnia/hypersomnia, and appetite loss/appetite gain were not predicted by any of the EMSs. Although the cross-sectional design of the study does not allow for conclusions about the direction of effects, results suggest that depression is not a unitary phenomenon and provide a possible explanation for previous inconsistent findings.


Wideband range-Doppler processing and beamforming using electro-optic arrays and spectral hole burning materials.
Ubiquitous radar systems look everywhere at all times and require both parallel radar processors and parallel beamformers. Current systems operate with subgigahertz bandwidths and produce a handful of angle-of-arrival (AOA) beams. We present an electro-optic radar processor that combines the multigigahertz wideband capabilities of a spectral hole burning correlator with wideband Doppler processing and the thousands of parallel channels available from an electro-optical beamformer. Preliminary experiments demonstrate 150 MHz bandwidth range correlations across 20 AOA beams.


Current therapeutic strategy in osteoarticular brucellosis.
Brucellosis is a common zoonotic disease with high morbidity. In the majority of human cases, the causative agent is B. melitensis. Infection is transmitted to humans by direct/indirect contact with the contaminated animal products (e.g., consumption of unpasteurized milk), infectious aerosols and aborted fetus. Brucellosis often affects middle-aged adults and young people. Patients with brucellosis tend to have non-specific symptoms, including fever, chills, night sweats, joint pain and myalgia. Brucellosis affects various organs and tissues. The osteoarticular system is one of the most commonly described affected systems in humans. In several clinical studies, the prevalence of Osteoarticular Brucellosis (OB) is reported as 2-77%. Most important osteoarticular clinical forms osteomyelitis, spondylitis, sacroiliitis, arthritis and bursitis. Spondylitis and spondylodiscitis are the most frequent complications. Spondylodiscitis often affects the lumbar (especially at the L4- L5 levels) and low thoracic vertebrae than the cervical spine. Back pain and sciatica radiculopathy are the most common complaints about patients. Sacroiliitis is associated with severe pain, especially back pain in affected individuals. Spinal destructive brucellar lesions are also reported in adults in previous studies. Brucellosis is diagnosed with clinical inflammatory signs (eg. tenderness, pain) of the affected joints together with positive serological tests and positive blood/synovial fluids cultures. Serological test measures the total amount of IgM/IgG antibodies. Standard agglutination test (SAT) titer ≥1:160 is in favor of brucellosis diagnosis. Enzyme-Linked Immunosorbent Assay (ELISA) and Polymerase chain reaction (PCR) are other types of diagnostic tests. Radiological assessments, such as joint sonography, computed tomography, magnetic resonance imaging, are the most helpful radiological methods to diagnose spinal brucellosis. The agents commonly used in the treatment of brucella spondylitis are doxycycline, streptomycin, gentamicin, ciprofloxacin, trimethoprim/sulfamethoxazole and rifampicin. The recommended regimens for treatment of brucella involve two or three antibiotics combinations. No standard treatment, physicians prescribe drugs based on conditions of the disease. Patients need a long-term (usually at three months) antibiotic therapy for mainly aiming to prevent relapses. Surgery may be required for patients with spinal abscess. This review focused on physicians' awareness for osteoarticular involvement, clinical presentation, diagnosis and current treatment of OB.


An evaluation of a chemical cautery agent and an anti-inflammatory ointment for the treatment of recurrent aphthous stomatitis: a pilot study.
Recurrent aphthous stomatitis is a very common condition, currently treated with anti-inflammatory agents, which palliate the symptoms. The purpose of this clinical trial was to compare a medication commonly used to treat recurrent aphthous stomatitis, Kenalog-in-Orabase, and a newer agent, Debacterol. Sixty patients diagnosed with recurrent aphthous stomatitis were enrolled in the study. Twenty patients were assigned to each of the two treatment groups, and 20 age- and sex-matched patients were assigned to the control group, which received no treatment. After the diagnosis was made, clinical examinations and ulcer measurements were performed, and a subjective evaluation of symptoms (100-mm visual analog scale) was completed by each subject. The subjects did not use any other medications. Both agents were applied topically (the frequency varied depending on the group of subjects) at specified intervals. Ulcer measurements and subjective evaluations were made at days 0, 3, 6, and 10 for all subjects. In both treatment groups, by day 10, 100% of the ulcers had clinically healed and were no longer causing pain. Patients in the Debacterol group reported a significantly greater decrease in pain at 3 days (> 70%) than did subjects in the other groups (< 20%), although the size of the ulcer did not differ significantly in any of the groups. After day 6, 80% of the ulcers in the Debacterol group had clinically disappeared and no longer caused symptoms, as compared to about 30% in the other groups. Patients subjectively reported significantly greater relief from symptoms with Debacterol than with Kenalog-in-Orabase or no treatment. The relief of symptoms associated with recurrent aphthous stomatitis may or may not correspond to clinical improvement, and these two topical medications may affect signs and symptoms of the lesions differently.


Seroepidemiological study of Toxoplasma gondii in equids in different European countries.
Toxoplasmosis, caused by the obligate intracellular protozoan Toxoplasma gondii, is a worldwide parasitic zoonosis. A cross-sectional study was carried out to determine the exposure to T. gondii in equids in Europe. Serum samples from 1399 equids (1085 horses, 238 donkeys, and 76 mules/hinnies) bred in four European countries (Italy, Spain, United Kingdom [UK], and Ireland) were collected during the period of 2013-2021. The overall seroprevalence of T. gondii was 18.9% (95% Confidence Interval [CI]: 16.9-21.0) by using the modified agglutination test (MAT) at a cut-off of 1:25. Seropositivity by country was 27.1% in Italy, 16.6% in Spain, 12.0% in UK and 7.0% in Ireland. Anti-T. gondii antibodies were detected in 12.8% of the horses, 43.7% of the donkeys, and in 28.9% of the mules/hinnies. A Generalized Estimating Equation (GEE) analysis was carried out to study the associations between seropositivity and explanatory variables related to individuals, herds, and management measures on these herds, selected based on the bivariate analysis. The risk for being seropositive for T. gondii was 5.3 and 2.7 times higher in donkeys and mules/hinnies than in horses, respectively. In addition, significantly higher seropositivity was observed in horses from herds that used disinfectants less than once a week (13.9%; p = 0.038, odds ratio [OR] = 1.6; 95% CI: 1.03-2.62) compared with those from herds that performed weekly disinfection of the facilities (9.4%). This is the first large-scale seroepidemiological study on T. gondii comprising horses, donkeys, and mules/hinnies in Europe and the first report of T. gondii exposure in horses from Ireland and UK. We found a widespread distribution of T. gondii among equid populations in different European countries. The seroprevalence found in these species, especially in donkeys and mules/hinnies, highlights the potential risk of human infection through the consumption of their raw/undercooked milk or meat.


Measurement of fast electron spin relaxation times with atomic resolution.
Single spins in solid-state systems are often considered prime candidates for the storage of quantum information, and their interaction with the environment the main limiting factor for the realization of such schemes. The lifetime of an excited spin state is a sensitive measure of this interaction, but extending the spatial resolution of spin relaxation measurements to the atomic scale has been a challenge. We show how a scanning tunneling microscope can measure electron spin relaxation times of individual atoms adsorbed on a surface using an all-electronic pump-probe measurement scheme. The spin relaxation times of individual Fe-Cu dimers were found to vary between 50 and 250 nanoseconds. Our method can in principle be generalized to monitor the temporal evolution of other dynamical systems.


Incidence of asymptomatic extracranial arterial disease.
Investigations of the incidence and the extent of the asymptomatic early stages of extracranial arterial disease (EAD) have been restricted for methodical reasons. Direct Continuous Wave-Doppler examination has given highly accurate results in the location and correct estimation of the degree of EAD both for the carotid (97%) and the vertebral arteries (90%), as shown from a detailed comparison with carotid (n = 604) and vertebral (n = 426) angiograms. Compared with this degree of reliability, the validity of normal auscultation for the diagnosis of EAD is shown to be poor: if bruits are taken as the only signs of associated EAD in patients with systemic atherosclerosis, only 27.6% in a group of 123 patients would have been correctly diagnosed. This parallels the number of false-positives (22.6%) in patients with normal results. The frequency and degree of EAD was studied by the use of direct Doppler examination in 2009 neurologically asymptomatic patients admitted either with severe vascular (n = 375) or coronary atherosclerosis (n = 262) or with high-risk factors (n = 1370). The frequency was significantly higher (32.8%) in patients with peripheral vascular disease than in those with coronary artery disease (6.8%) and in risk-factor patients (5.9%). The combination and degree of vessel involvement are presented in detail and their possible prognostic significance discussed.


SchNet - A deep learning architecture for molecules and materials.
Deep learning has led to a paradigm shift in artificial intelligence, including web, text, and image search, speech recognition, as well as bioinformatics, with growing impact in chemical physics. Machine learning, in general, and deep learning, in particular, are ideally suitable for representing quantum-mechanical interactions, enabling us to model nonlinear potential-energy surfaces or enhancing the exploration of chemical compound space. Here we present the deep learning architecture SchNet that is specifically designed to model atomistic systems by making use of continuous-filter convolutional layers. We demonstrate the capabilities of SchNet by accurately predicting a range of properties across chemical space for molecules and materials, where our model learns chemically plausible embeddings of atom types across the periodic table. Finally, we employ SchNet to predict potential-energy surfaces and energy-conserving force fields for molecular dynamics simulations of small molecules and perform an exemplary study on the quantum-mechanical properties of C20-fullerene that would have been infeasible with regular ab initio molecular dynamics.


Collagen-induced arthritis in common marmosets: a new nonhuman primate model for chronic arthritis.
There is an ever-increasing need for animal models to evaluate efficacy and safety of new therapeutics in the field of rheumatoid arthritis (RA). Particularly for the early preclinical evaluation of human-specific biologicals targeting the progressive phase of the disease, there is a need for relevant animal models. In response to this requirement we set out to develop a model of collagen-induced arthritis (CIA) in a small-sized nonhuman primate species (300 to 400 g at adult age); that is, the common marmoset (Callithrix jacchus). Twenty-two animals divided into three experiments were immunized with collagen type II (CII) of either bovine or chicken origin with different immunization strategies. The animals were analyzed for clinical manifestation of arthritis, hematology and clinical chemistry, immunological responses against CII and histopathological features of the arthritis. Clinically manifest arthritis was observed in almost 100% (21 out of 22) of the animals. Fifty percent of the animals developed semi-acute CIA while the other 50% displayed a more chronic disease. Both cellular (CD3/CD4 and CD3/CD8) and humoral responses (IgM and IgG) against CII were involved in the development of the disease. Besides mild histopathological changes in bone and cartilage, severe inflammation in extraarticular tissues like periosteum and subcutaneous tissues was observed. This new model in marmosets more closely resembles chronic RA with respect to the chronic disease course and pathomorphological presentation than the more acute monophasic and destructive CIA model in macaques. This model can therefore fill a niche in preclinical testing of new human specific therapeutics.


Secretion of d-alanine by Escherichia coli.
Escherichia coli has an l-alanine export system that protects the cells from toxic accumulation of intracellular l-alanine in the presence of l-alanyl-l-alanine (l-Ala-l-Ala). When a DadA-deficient strain was incubated with 6.0 mM l-Ala-l-Ala, we detected l-alanine and d-alanine using high-performance liquid chromatography (HPLC) analysis at a level of 7.0 mM and 3.0 mM, respectively, after 48 h incubation. Treatment of the culture supernatant with d-amino acid oxidase resulted in the disappearance of a signal corresponding to d-alanine. Additionally, the culture supernatant enabled a d-alanine auxotroph to grow without d-alanine supplementation, confirming that the signal detected by HPLC was authentic d-alanine. Upon introduction of an expression vector harbouring the alanine racemase genes, alr or dadX, the extracellular level of d-alanine increased to 11.5 mM and 8.5 mM, respectively, under similar conditions, suggesting that increased metabolic flow from l-alanine to d-alanine enhanced d-alanine secretion. When high-density DadA-deficient cells preloaded with l-Ala-l-Ala were treated with 20 µM carbonyl cyanide m-chlorophenyl hydrazone (CCCP), secretion of both l-alanine and d-alanine was enhanced ~twofold compared with that in cells without CCCP treatment. In contrast, the ATPase inhibitor dicyclohexylcarbodiimide did not exert such an effect on the l-alanine and d-alanine secretion. Furthermore, inverted membrane vesicles prepared from DadA-deficient cells lacking the l-alanine exporter AlaE accumulated [3H]D-alanine in an energy-dependent manner. This energy-dependent accumulation of [3H]D-alanine was strongly inhibited by CCCP. These results indicate that E. coli has a transport system(s) that exports d-alanine and that this function is most likely modulated by proton electrochemical potential.


The protective role of Nrf2 in streptozotocin-induced diabetic nephropathy.
Diabetic nephropathy is one of the major causes of renal failure, which is accompanied by the production of reactive oxygen species (ROS). Nrf2 is the primary transcription factor that controls the antioxidant response essential for maintaining cellular redox homeostasis. Here, we report our findings demonstrating a protective role of Nrf2 against diabetic nephropathy. We explore the protective role of Nrf2 against diabetic nephropathy using human kidney biopsy tissues from diabetic nephropathy patients, a streptozotocin-induced diabetic nephropathy model in Nrf2(-/-) mice, and cultured human mesangial cells. The glomeruli of human diabetic nephropathy patients were under oxidative stress and had elevated Nrf2 levels. In the animal study, Nrf2 was demonstrated to be crucial in ameliorating streptozotocin-induced renal damage. This is evident by Nrf2(-/-) mice having higher ROS production and suffering from greater oxidative DNA damage and renal injury compared with Nrf2(+/+) mice. Mechanistic studies in both in vivo and in vitro systems showed that the Nrf2-mediated protection against diabetic nephropathy is, at least, partially through inhibition of transforming growth factor-beta1 (TGF-beta1) and reduction of extracellular matrix production. In human renal mesangial cells, high glucose induced ROS production and activated expression of Nrf2 and its downstream genes. Furthermore, activation or overexpression of Nrf2 inhibited the promoter activity of TGF-beta1 in a dose-dependent manner, whereas knockdown of Nrf2 by siRNA enhanced TGF-beta1 transcription and fibronectin production. This work clearly indicates a protective role of Nrf2 in diabetic nephropathy, suggesting that dietary or therapeutic activation of Nrf2 could be used as a strategy to prevent or slow down the progression of diabetic nephropathy.


Lumican is required for neutrophil extravasation following corneal injury and wound healing.
An important aspect of wound healing is the recruitment of neutrophils to the site of infection or tissue injury. Lumican, an extracellular matrix component belonging to the small leucine rich proteoglycan (SLRP) family, is one of the major keratan sulfate proteoglycans (KSPGs) within the corneal stroma. Increasing evidence indicates that lumican can serve as a regulatory molecule for several cellular processes, including cell proliferation and migration. In the present study, we addressed the role of lumican in the process of extravasation of polymorphonuclear leukocytes (PMNs) during the early inflammatory phase present in the healing of the corneal epithelium following debridement. We used Lum(-/-) mice and a novel transgenic mouse, Lum(-/-),Kera-Lum, which expresses lumican only in the corneal stroma, to assess the role of lumican in PMN extravasation into injured corneas. Our results showed that PMNs did not readily invade injured corneas of Lum(-/-) mice and this defect was rescued by the expression of lumican in the corneas of Lum(-/-),Kera-Lum mice. The presence of lumican in situ facilitates PMN infiltration into the peritoneal cavity in casein-induced inflammation. Our findings are consistent with the notion that in addition to regulating the collagen fibril architecture, lumican acts to aid neutrophil recruitment and invasion following corneal damage and inflammation.


Hypolipidemic influence of dietary fenugreek (Trigonella foenum-graecum) seeds and garlic (Allium sativum) in experimental myocardial infarction.
The cardioprotective influence of dietary fibre-rich fenugreek seeds and the well-established hypolipidemic spice garlic was evaluated both individually and in combination in isoproterenol induced myocardial infarcted rats. It was particularly examined whether pretreatment with dietary fenugreek, garlic or fenugreek + garlic would be beneficial under hypercholesterolemic conditions by their influence on the tissue lipid profile. Four groups each of male Wistar rats were maintained on either a basal diet or a high cholesterol diet for 8 weeks. Dietary interventions with fenugreek, garlic and the combination of fenugreek and garlic were made by including 10% fenugreek seed powder, 2% freeze-dried garlic powder, and 10% fenugreek seed powder + 2% garlic powder. At the end of the diet regimen, myocardial infarction was induced with isoproterenol (i.p. 80 mg kg(-1)) twice at intervals of 12 h. The disturbed activities of cardiac marker enzymes in serum and the heart confirmed isoproterenol induced myocardial infarction. Dietary fenugreek, garlic or fenugreek + garlic was found to ameliorate the pathological changes in heart tissue and lipid abnormalities in serum and the heart, the beneficial effect being higher with the combination of fenugreek and garlic, invariably amounting to an additive effect. The results also indicated that the hypercholesterolemic situation aggravated the myocardial damage during isoproterenol-induced myocardial infarction. This dietary intervention study suggested that the combination of fenugreek seeds and garlic offers a higher beneficial influence in exerting the cardioprotective effect.


Clinical research on the therapeutic effect of the electro-acupuncture treatment in patients with depression.
Electroacupuncture (EA) stimulation has been found to influence the brain (norepinephrine metabolism in experimental animals). Preliminary clinical research has shown that EA treatment is as effective as amitriptyline for patients with depression. In this study, two consecutive clinical studies on the treatment of depression with EA are conducted. The first study was double blind placebo controlled, in which 29 depressed inpatients were recruited. Patients were randomly divided into three groups: EA + placebo; amitriptyline; and EA + amitriptyline. They received EA and/or amitriptyline treatment for 6 weeks. The Hamilton Rating Scale for Depression, Clinical Global Impression and ASBERG scales for the side effect of antidepressants were used to evaluate the therapeutic efficacy and side effects. Based on the results and research protocol of the first study, a multi-centered collaborative study was conducted, in which 241 inpatients with depression were recruited. Patients were randomly divided into two treatment groups: the EA + placebo and the amitriptyline groups. The results from both studies showed that the therapeutic efficacy of EA was equal to that of amitriptyline for depressive disorders (P > 0.05). Electro-acupuncture had a better therapeutic effect for anxiety somatization and cognitive process disturbance of depressed patients than amitriptyline (P < 0.05). Moreover, the side effects of EA were much less than that of amitriptyline (P < 0.001). The article suggested that EA treatment was an effective therapeutic method for depressive disorders. Particularly, it was a treatment of choice for depressed patients who were unable to comply with the classic tricyclic antidepressants because of their anticholinergic side effects. The possible mechanism of EA treatment is discussed.


Effects of chlordiazepoxide on extinction and re-acquisition of operant behaviour in mice.
Relatively little is known about the role of the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) in extinction of appetitively motivated tasks. The benzodiazepine (BZ) chlordiazepoxide (CDP) was administered during extinction and re-acquisition of lever pressing by mice following food reinforced discrete-trial fixed-ratio 5 (FR-5) training. Typical FR behaviour was established during baseline training and persisted for several extinction sessions. There were 15 extinction sessions in all, followed by six re-acquisition sessions where food reinforcement was re-introduced. In a 2x2x2 between-group design, CDP (15 mg/kg) or vehicle injections were given prior to either the last two food reinforcement sessions and the first 10 extinction sessions, or the final five extinction sessions, or the six re-acquisition sessions. Initially CDP had no effect on the rate of extinction, but after several extinction sessions it significantly facilitated it. Surprisingly, if CDP was administered only after several sessions of extinction, it immediately produced facilitation. Thus the delayed effects of CDP are not due to drug accumulation. These data suggest that some neural change must occur before CDP can affect extinction processes. In re-acquisition sessions, CDP facilitated the reinstatement of food-reinforced lever pressing. Implications for neural and behavioural accounts of operant extinction are discussed.


Solid-state colloidal CuInS2 quantum dot solar cells enabled by bulk heterojunctions.
Colloidal copper indium sulfide (CIS) nanocrystals (NCs) are Pb- and Cd-free alternatives for use as absorbers in quantum dot solar cells. In a heterojunction with TiO2, non-annealed ligand-exchanged CIS NCs form solar cells yielding a meager power conversion efficiency (PCE) of 0.15%, with photocurrents plummeting far below predicted values from absorption. Decreasing the amount of zinc during post-treatment leads to improved mobility but marginal improvement in device performance (PCE = 0.30%). By incorporating CIS into a porous TiO2 network, we saw an overall drastic improvement in device performance, reaching a PCE of 1.16%, mainly from an increase in short circuit current density (Jsc) and fill factor (FF) and a 10-fold increase in internal quantum efficiency (IQE). We have determined that by moving from a bilayer to a bulk heterojunction architecture, we have reduced the trap-assisted recombination as seen in changes in the ideality factor, the intensity dependence of the photocurrent and transient photocurrent (TPC) and photovoltage (TPV) characteristics.


Laparotomy for blunt abdominal trauma in a civilian trauma service.
This report looks at the group of patients who required a laparotomy for blunt torso trauma at a busy metropolitan trauma service in South Africa. Methods. A prospective trauma registry is maintained by the surgical services of the Pietermaritzburg metropolitan complex. This registry is interrogated retrospectively. All patients who required admission for blunt torso trauma over the period September 2006 - September 2007 were included for review. Proformas documenting mechanism of injury, age, vital signs, blood gas, delay in presentation, length of hospital stay, intensive care unit stay and operative details were completed. Results. A total of 926 patients were treated for blunt trauma by the Pietermaritzburg metropolitan services during the period under consideration. A cohort of 65 (8%) required a laparotomy for blunt trauma during this period. There were 17 females in this group. The mechanisms of injury were motor vehicle accident (MVA) (27), pedestrian vehicle accident (PVA) (21), assault (5), fall from a height (3), bicycle accident (6), quad bike accident (1) and tractor-related accident (2). The following isolated injuries were discovered at laparotomy: liver (9), spleen (5), diaphragm (1), duodenum (2), small bowel (8), mesentery (8) bladder (10), gallbladder (1), stomach (2), colon/rectum (2) and retrohepatic vena cava (1). The following combined injuries were discovered: liver and diaphragm (2), spleen and pancreas (1), spleen and liver (2), spleen, aorta and diaphragm (1), spleen and bladder (1) and small bowel and bladder (2). Eighteen patients in the series (26%) required relaparotomy. In 10 patients temporary abdominal containment was needed. The mortality rate was 26% (18 patients). There were 6 deaths from massive bleeding, all within 6 hours of operation, and 3 deaths from renal failure; the remaining 9 patients died of multiple organ failure. There were 8 negative laparotomies (7%). In the negative laparotomy group false-positive computed tomography (CT) scan findings were a problem in 3 cases, in 1 case hypotension and a fractured pelvis on admission prompted laparotomy, and in the other cases clinical findings prompted laparotomy. All patients who underwent negative laparotomy survived. There were 10 pelvic fractures, 5 lower limb fractures, 2 spinal injuries, 4 femur fractures and 2 upper limb fractures. CT scans were done in 25 patients. In 20 patients the systolic blood pressure on presentation was <90 mmHg and in 41 the pulse rate was >110 beats/min. In 16 patients there was a base excess of <-4 on presentation. Conclusion. Laparotomy is needed in less than 10% of patients who sustain blunt abdominal trauma. Solid visceral injury requiring laparotomy presents with haemodynamic instability. Hollow visceral injury has a more insidious presentation and is associated with a delay in diagnosis. CT scan is the most widely used investigation in blunt abdominal trauma. It is both sensitive and specific for solid visceral injury, but its accuracy for the diagnosis of hollow visceral injury is less well defined. Clinical suspicion must be high, and hollow visceral injury needs to be actively excluded.


Factors controlling oxygen utilization.
We demonstrate, theoretically, that oxygen diffusion distance is related to the metabolic rate of tumors (QO2) as well as the oxygen tension. The difference in QO2 rate between tumors can vary by as much as 80-fold. Inhibition of oxygen utilization by glucose or chemical inhibitors can improve the diffusion distance. Combining respiratory inhibitors with increased availability of oxygen will further improve the oxygen diffusion distance for all tumors. A simple means for inhibiting oxygen consumption is the use of glucose (the Crabtree effect). The inhibition of tumor oxygen utilization by glucose occurs in R323OAc mammary carcinoma and 9L glioma cells. However, stimulation of oxygen consumption is observed with glucose in the Q7 hepatoma cell line. MIBG, a known inhibitor of oxygen utilization, blocks oxygen consumption in 9L, but is weakly inhibitory with the Q7. Q7 tumor cells demonstrate an anomalous behavior of glucose and MIBG on oxygen consumption. Our results clearly demonstrate the necessity for comparing effects of different agents on different tumor cells. Generalizations cannot be made with respect to the choice of inhibitor for in vivo use. Our work shows that oxygen consumption also can be inhibited with malonate and chlorosuccinate. These substrates may be effective in vivo, where glucose is low and glutamine is the major substrate. Our results indicate that information about individual tumor substrate-linked metabolic controls may be necessary before attempting to inhibit oxygen utilization in vivo for therapeutic benefit.


AMPA receptors regulate experience-dependent dendritic arbor growth in vivo.
The size and shape of neuronal dendritic arbors affect the number and type of synaptic inputs, as well as the complexity and function of brain circuits. In the intact brain, dendritic arbor growth and the development of excitatory glutamatergic synapse are concurrent. Consequently, it has been difficult to resolve whether synaptic inputs drive dendritic arbor development. Here, we test the role of AMPA receptor (AMPAR)-mediated glutamatergic transmission in dendrite growth by expressing peptides corresponding to the intracellular C-terminal domains of AMPAR subunits GluR1 (GluR1Ct) and GluR2 (GluR2Ct) in optic tectal neurons of the Xenopus retinotectal system. These peptides significantly reduce AMPAR synaptic transmission in transfected neurons while leaving visual system circuitry intact. Daily in vivo imaging over 5 days revealed that GluR1Ct or GluR2Ct expression dramatically impaired dendrite growth, resulting in less complex arbors than controls. Time-lapse images collected at 2-h intervals over 6 h show that both GluR1Ct and GluR2Ct decrease branch lifetimes. Ultrastructural analysis indicates that synapses formed onto neurons expressing the GluRCt are less mature than synapses onto control neurons. These data suggest that the failure to form complex arbors is due to reduced stabilization of new synapses and dendritic branches. Although visual stimulation increases dendritic arbor growth rates in control tectal neurons, a weak postsynaptic response to visual experience in GluRCt-expressing cells leads to retraction of branches. These results indicate that AMPAR-mediated transmission underlies experience-dependent dendritic arbor growth by stabilizing branches, and support a competition-based model for dendrite growth.


Homodimerization of amyloid precursor protein and its implication in the amyloidogenic pathway of Alzheimer's disease.
We reported previously that the carbohydrate domain of the amyloid precursor protein is involved in amyloid precursor protein (APP)-APP interactions. Functional in vitro studies suggested that this interaction occurs through the collagen binding site of APP. The physiological significance remained unknown, because it is not understood whether and how APP dimerization occurs in vivo. Here we report that cellular APP exists as homodimers matching best with a two-site model. Consistent with our published crystallographic data, we show that a deletion of the entire sequence after the kunitz protease inhibitor domain did not abolish APP homodimerization, suggesting that two domains are critically involved but that neither is essential for homodimerization. Finally, we generated stabilized dimers by expressing mutant APP with a single cysteine in the ectodomain juxtamembrane region. Mutation of Lys(624) to cysteine produced approximately 6-8-fold more A beta than cells expressing normal APP. Our results suggest that amyloid A beta production can in principle be positively regulated by dimerization in vivo. We suggest that dimerization could be a physiologically important mechanism for regulating the proposed signal activity of APP.


International children's accelerometry database (ICAD): design and methods.
Over the past decade, accelerometers have increased in popularity as an objective measure of physical activity in free-living individuals. Evidence suggests that objective measures, rather than subjective tools such as questionnaires, are more likely to detect associations between physical activity and health in children. To date, a number of studies of children and adolescents across diverse cultures around the globe have collected accelerometer measures of physical activity accompanied by a broad range of predictor variables and associated health outcomes. The International Children's Accelerometry Database (ICAD) project pooled and reduced raw accelerometer data using standardized methods to create comparable outcome variables across studies. Such data pooling has the potential to improve our knowledge regarding the strength of relationships between physical activity and health. This manuscript describes the contributing studies, outlines the standardized methods used to process the accelerometer data and provides the initial questions which will be addressed using this novel data repository. Between September 2008 and May 2010 46,131 raw Actigraph data files and accompanying anthropometric, demographic and health data collected on children (aged 3-18 years) were obtained from 20 studies worldwide and data was reduced using standardized analytical methods. When using ≥ 8, ≥ 10 and ≥ 12 hrs of wear per day as a criterion, 96%, 93.5% and 86.2% of the males, respectively, and 96.3%, 93.7% and 86% of the females, respectively, had at least one valid day of data. Pooling raw accelerometer data and accompanying phenotypic data from a number of studies has the potential to: a) increase statistical power due to a large sample size, b) create a more heterogeneous and potentially more representative sample, c) standardize and optimize the analytical methods used in the generation of outcome variables, and d) provide a means to study the causes of inter-study variability in physical activity. Methodological challenges include inflated variability in accelerometry measurements and the wide variation in tools and methods used to collect non-accelerometer data.


The RNA-binding protein KSRP promotes the biogenesis of a subset of microRNAs.
Consistent with the role of microRNAs (miRNAs) in down-regulating gene expression by reducing the translation and/or stability of target messenger RNAs, the levels of specific miRNAs are important for correct embryonic development and have been linked to several forms of cancer. However, the regulatory mechanisms by which primary miRNAs (pri-miRNAs) are processed first to precursor miRNAs (pre-miRNAs) and then to mature miRNAs by the multiprotein Drosha and Dicer complexes, respectively, remain largely unknown. The KH-type splicing regulatory protein (KSRP, also known as KHSRP) interacts with single-strand AU-rich-element-containing mRNAs and is a key mediator of mRNA decay. Here we show in mammalian cells that KSRP also serves as a component of both Drosha and Dicer complexes and regulates the biogenesis of a subset of miRNAs. KSRP binds with high affinity to the terminal loop of the target miRNA precursors and promotes their maturation. This mechanism is required for specific changes in target mRNA expression that affect specific biological programs, including proliferation, apoptosis and differentiation. These findings reveal an unexpected mechanism that links KSRP to the machinery regulating maturation of a cohort of miRNAs that, in addition to its role in promoting mRNA decay, independently serves to integrate specific regulatory programs of protein expression.


Distribution of pulmonary ventilation and perfusion measured simultaneously in awake goats.
Gravity has been regarded as the major determinant for local pulmonary perfusion and ventilation. Recent reports, describing major gravity independent heterogeneity in both variables, have questioned the importance of gravity. We asked to what extent ventilation and perfusion were related, and if they showed similar distributions along the vertical axis in the lung. We gave 99mTc-aerosols as tracers for ventilation and radioactive microspheres as blood flow tracers in five awake goats over 4 min. Ventilation and perfusion were determined in approximately 1.5 cm3 pieces of the lung. For both variables the vertical distribution could vary considerably from lung to lung, but within each lung the two distributions were similar. Both ventilation and perfusion were heterogeneously distributed (CV approximately 40% for both), they were highly correlated (r = 0.81) and the average 25-75-interpercentile interval for ventilation to perfusion ratio (0.84-1.13) was significantly less wide than for both ventilation (0.76-1.38) and perfusion (0.76-1.40). Some pieces were considerably overventilated while a few were correspondingly underventilated. This could indicate that perfusion is adjusted to ventilation in normoxic lungs with a low sensitivity to overventilation.


TMS over the supramarginal gyrus delays selection of appropriate grasp orientation during reaching and grasping tools for use.
Tool use, a ubiquitous part of human behaviour, requires manipulation control and knowledge of tool purpose. Neuroimaging and neuropsychological research posit that these two processes are supported by separate brain regions, ventral premotor and inferior parietal for manipulation control, and posterior middle temporal cortex for tool knowledge, lateralised to the left hemisphere. Action plans for tool use need to integrate these two separate processes, which is likely supported by the left supramarginal gyrus (SMG). However, whether this integration occurs during action execution is not known. To clarify the role of the SMG we conducted two experiments in which healthy participants reached to grasp everyday tools with the explicit instruction to use them directly following their grasp. To study the integration of manipulation control and tool knowledge within a narrow time window we mechanically perturbed the orientation of the tool to force participants to correct grasp orientation 'on-line' during the reaching movement. In experiment 1, twenty healthy participants reached with their left hand to grasp a tool. Double-pulse transcranial magnetic stimulation (TMS) was applied, in different blocks over left or right SMG at the onset of perturbation. Kinematic data revealed delayed and erroneous online correction after TMS over left and right SMG. In Experiment 2 twelve participants reached, in different blocks, with their left or right hand and TMS was applied over SMG ipsilateral to the reaching hand. A similar effect on correction was observed for ipsilateral stimulation when reaching with the left and right hands, and no effect of or interaction with hemisphere was observed. Our findings implicate a bilateral role of the SMG in correcting movements and selection of appropriate grasp orientation during reaching to grasp tools for use.


Functional Connectivity under Anticipation of Shock: Correlates of Trait Anxious Affect versus Induced Anxiety.
Sustained anxiety about potential future negative events is an important feature of anxiety disorders. In this study, we used a novel anticipation of shock paradigm to investigate individual differences in functional connectivity during prolonged threat of shock. We examined the correlates of between-participant differences in trait anxious affect and induced anxiety, where the latter reflects changes in self-reported anxiety resulting from the shock manipulation. Dissociable effects of trait anxious affect and induced anxiety were observed. Participants with high scores on a latent dimension of anxious affect showed less increase in ventromedial pFC-amygdala connectivity between periods of safety and shock anticipation. Meanwhile, lower levels of induced anxiety were linked to greater augmentation of dorsolateral pFC-anterior insula connectivity during shock anticipation. These findings suggest that ventromedial pFC-amygdala and dorsolateral pFC-insula networks might both contribute to regulation of sustained fear responses, with their recruitment varying independently across participants. The former might reflect an evolutionarily old mechanism for reducing fear or anxiety, whereas the latter might reflect a complementary mechanism by which cognitive control can be implemented to diminish fear responses generated due to anticipation of aversive stimuli or events. These two circuits might provide complementary, alternate targets for exploration in future pharmacological and cognitive intervention studies.


Paracrine immunotherapy with interleukin-2 and local chemotherapy is synergistic in the treatment of experimental brain tumors.
Potent immune responses against malignant brain tumors can be elicited by paracrine intracranial (i.c.) immunotherapy with interleukin (IL)-2. Additionally, i.c. delivery of carmustine via biodegradable polymers has been shown to significantly prolong survival in both animal models and clinical trials. In this study, we show that the combination of paracrine immunotherapy, with nonreplicating genetically engineered tumor cells that produce IL-2, and local delivery of chemotherapy by biodegradable polymers prolongs survival in a synergistic manner in mice challenged intracranially with a lethal murine brain tumor. Animals receiving IL-2-transduced cells and polymers containing 10% 1,3-bis(2-chloroethyl)-1-nitrosourea had significantly improved survival compared with animals receiving IL-2-transduced cells or 10% 1,3-bis(2-chloroethyl)-1-nitrosourea alone. Median survival for the control group was 19 days. Survival in animals receiving IL-2-transduced cells and 1% carboplatin-containing polymers was also significantly improved compared with either therapy alone. Histopathological examination on day 14 of animals receiving combination treatment showed rare degenerating tumor cells. In addition to tissue necrosis surrounding the polymer, a marked inflammatory reaction was observed. In long-term survivors (all animals receiving combination treatment), no tumor was observed and the inflammatory reaction was completely resolved. The brains of animals receiving combination therapy showed both tissue necrosis due to local chemotherapy and strong inflammation due to paracrine immunotherapy. The demonstration of synergy between paracrine IL-2 and local i.c. delivery of antineoplastic drugs is novel and may provide a combined treatment strategy for use against both primary and metastatic i.c. tumors.


Relationship between gastric emptying of solids and gall bladder emptying in normal subjects.
Very little is known about the normal temporal and quantitative relationships between gastric emptying and gall bladder emptying. Using a non-invasive double isotope technique these relationships were investigated in 22 normal healthy adults. 99Tcm EHIDA was used as the biliary tracer and 113Inm labelled bran as the gastric content tracer. Gastric emptying was monoexponential with a t1/2 of 45 +/- 3 minutes (mean +/- SEM). In 15 subjects the gall bladder emptied in relation to eating according to a double exponential function. In these subjects 15.0 +/- 1.6% of gall bladder contents emptied before gastric emptying began. They could be further divided into two clear cut types (p less than 0.001), according to the ejection fraction at 10 minutes and the t1/2 of the first exponential. Emptying of the gall bladder was faster and more of its contents were ejected in subjects with a type I response (n = 9) than in subjects with a type II response (n = 6). In the remaining seven subjects the gall bladder began to empty spontaneously, unrelated to eating. These observations suggest that gall bladder emptying: (a) may have a cephalic phase, (b) can be expressed as a double exponential function, (c) may occur unrelated to eating, (d) which occurs only in relation to eating would appear to be either fast (type I) or slow (type II).


3R measures in facilities for the production of genetically modified rodents.
Sociocultural changes in the human-animal relationship have led to increasing demands for animal welfare in biomedical research. The 3R concept is the basis for bringing this demand into practice: Replace animal experiments with alternatives where possible, Reduce the number of animals used to a scientifically justified minimum and Refine the procedure to minimize animal harm. The generation of gene-modified sentient animals such as mice and rats involves many steps that include various forms of manipulation. So far, no coherent analysis of the application of the 3Rs to gene manipulation has been performed. Here we provide guidelines from the Committee on Genetics and Breeding of Laboratory Animals of the German Society for Laboratory Animal Science to implement the 3Rs in every step during the generation of genetically modified animals. We provide recommendations for applying the 3Rs as well as success/intervention parameters for each step of the process, from experiment planning to choice of technology, harm-benefit analysis, husbandry conditions, management of genetically modified lines and actual procedures. We also discuss future challenges for animal welfare in the context of developing technologies. Taken together, we expect that our comprehensive analysis and our recommendations for the appropriate implementation of the 3Rs to technologies for genetic modifications of rodents will benefit scientists from a wide range of disciplines and will help to improve the welfare of a large number of laboratory animals worldwide.


Familial diffuse cortical dysplasia.
Two brothers are described with a syndrome characterized by mental retardation, atypical absence, atonic and generalized tonic-clonic seizures, and bilateral symmetrical slow spike and wave discharges on electroencephalograms. Magnetic resonance imaging revealed dysplastic cortex probably representing pachygyria primarily over the parietal regions bilaterally in both patients. The presence of an identical clinical, electroencephalographic, and cortical developmental abnormality on imaging studies in these brothers suggests that a genetic mechanism may play a role in some developmental disorders of the cerebral cortex.


Self-esteem and peer-perceived social status in early adolescence and prediction of eating pathology in young adulthood.
Self-esteem is implied as a factor in the development of eating disorders. In adolescence peers have an increasing influence. Support for the role of self-esteem in eating disorders is ambiguous and little is known about the influence of social status as judged by others. The present study investigates whether self-esteem and peer status in early adolescence are associated with eating pathology in young adulthood. This study is part of TRAILS, a longitudinal cohort study on mental health and social development from preadolescence into adulthood. At age 11, participants completed the Self-Perception Profile for Children, assessing global self-esteem and self-perceptions regarding social acceptance, physical appearance, and academic competence. At age 13, peer status among classmates was assessed regarding likeability, physical attractiveness, academic performance, and popularity in a subsample of 1,007 participants. The Eating Disorder Diagnostic Scale was administered at age 22. The present study included peer-nominated participants with completed measures of self-perception at age 11 and eating pathology at age 22 (N = 732; 57.8% female). In a combined model, self-perceived physical attractiveness at age 11 and peer popularity at age 13 were inversely correlated with eating pathology at 22 years, while likeability by peers at age 13 was positively related to eating pathology. Both self-perceptions and peer status in early adolescence are significant predictors of eating pathology in young adults. Specific measures of self-esteem and peer-perceived status may be more relevant to the prediction of eating pathology than a global measure of self-esteem.


Palladium and platinum nanoparticles attenuate aging-like skin atrophy via antioxidant activity in mice.
Cu-Zn superoxide dismutase (Sod1) loss causes a redox imbalance as it leads to excess superoxide generation, which results in the appearance of various aging-related phenotypes, including skin atrophy. Noble metal nanoparticles, such as palladium (Pd) and platinum (Pt) nanoparticles, are considered to function as antioxidants due to their strong catalytic activity. In Japan, a mixture of Pd and Pt nanoparticles called PAPLAL has been used to treat chronic diseases over the past 60 years. In the present study, we investigated the protective effects of PAPLAL against aging-related skin pathologies in mice. Transdermal PAPLAL treatment reversed skin thinning associated with increased lipid peroxidation in Sod1-/- mice. Furthermore, PAPLAL normalized the gene expression levels of Col1a1, Mmp2, Has2, Tnf-α, Il-6, and p53 in the skin of the Sod1-/- mice. Pt nanoparticles exhibited marked SOD and catalase activity, while Pd nanoparticles only displayed weak SOD and catalase activity in vitro. Although the SOD and catalase activity of the Pt nanoparticles significantly declined after they had been oxidized in air, a mixture of Pd and Pt nanoparticles continued to exhibit SOD and catalase activity after oxidation. Importantly, a mixture of Pd and Pt nanoparticles with a molar ratio of 3 or 4 to 1 continued to exhibit SOD and catalase activity after oxidation, indicating that Pd nanoparticles prevent the oxidative deterioration of Pt nanoparticles. These findings indicate that PAPLAL stably suppresses intrinsic superoxide generation both in vivo and in vitro via SOD and catalase activity. PAPLAL is a potentially powerful tool for the treatment of aging-related skin diseases caused by oxidative damage.


Augmented uterine artery blood flow and oxygen delivery protect Andeans from altitude-associated reductions in fetal growth.
The effect of high altitude on reducing birth weight is markedly less in populations of high- (e.g., Andeans) relative to low-altitude origin (e.g., Europeans). Uterine artery (UA) blood flow is greater during pregnancy in Andeans than Europeans at high altitude; however, it is not clear whether such blood flow differences play a causal role in ancestry-associated variations in fetal growth. We tested the hypothesis that greater UA blood flow contributes to the protection of fetal growth afforded by Andean ancestry by comparing UA blood flow and fetal growth throughout pregnancy in 137 Andean or European residents of low (400 m; European n = 28, Andean n = 23) or high (3,100-4,100 m; European n = 51, Andean n = 35) altitude in Bolivia. Blood flow and fetal biometry were assessed by Doppler ultrasound, and maternal ancestry was confirmed, using a panel of 100 ancestry-informative genetic markers (AIMs). At low altitude, there were no ancestry-related differences in the pregnancy-associated rise in UA blood flow, fetal biometry, or birth weight. At high altitude, Andean infants weighed 253 g more than European infants after controlling for gestational age and other known influences. UA blood flow and O(2) delivery were twofold greater at 20 wk in Andean than European women at high altitude, and were paralleled by greater fetal size. Moreover, variation in the proportion of Indigenous American ancestry among individual women was positively associated with UA diameter, blood flow, O(2) delivery, and fetal head circumference. We concluded that greater UA blood flow protects against hypoxia-associated reductions in fetal growth, consistent with the hypothesis that genetic factors enabled Andeans to achieve a greater pregnancy-associated rise in UA blood flow and O(2) delivery than European women at high altitude.


An experimental method for the assessment of color simulation tools.
The Simulcheck method for evaluating the accuracy of color simulation tools in relation to dichromats is described and used to test three color simulation tools: Variantor, Coblis, and Vischeck. A total of 10 dichromats (five protanopes, five deuteranopes) and 10 normal trichromats participated in the current study. Simulcheck includes two psychophysical tasks: the Pseudoachromatic Stimuli Identification task and the Minimum Achromatic Contrast task. The Pseudoachromatic Stimuli Identification task allows determination of the two chromatic angles (h(uv) values) that generate a minimum response in the yellow–blue opponent mechanism and, consequently, pseudoachromatic stimuli (greens or reds). The Minimum Achromatic Contrast task requires the selection of the gray background that produces minimum contrast (near zero change in the achromatic mechanism) for each pseudoachromatic stimulus selected in the previous task (L(R) values). Results showed important differences in the colorimetric transformations performed by the three evaluated simulation tools and their accuracy levels. Vischeck simulation accurately implemented the algorithm of Brettel, Viénot, and Mollon (1997). Only Vischeck appeared accurate (similarity in huv and L(R) values between real and simulated dichromats) and, consequently, could render reliable color selections. It is concluded that Simulcheck is a consistent method because it provided an equivalent pattern of results for huv and L(R) values irrespective of the stimulus set used to evaluate a simulation tool. Simulcheck was also considered valid because real dichromats provided expected huv and LR values when performing the two psychophysical tasks included in this method.


Obesity Drives Delayed Infarct Expansion, Inflammation, and Distinct Gene Networks in a Mouse Stroke Model.
Obesity is associated with chronic peripheral inflammation, is a risk factor for stroke, and causes increased infarct sizes. To characterize how obesity increases infarct size, we fed a high-fat diet to wild-type C57BL/6J mice for either 6 weeks or 15 weeks and then induced distal middle cerebral artery strokes. We found that infarct expansion happened late after stroke. There were no differences in cortical neuroinflammation (astrogliosis, microgliosis, or pro-inflammatory cytokines) either prior to or 10 h after stroke, and also no differences in stroke size at 10 h. However, by 3 days after stroke, animals fed a high-fat diet had a dramatic increase in microgliosis and astrogliosis that was associated with larger strokes and worsened functional recovery. RNA sequencing revealed a dramatic increase in inflammatory genes in the high-fat diet-fed animals 3 days after stroke that were not present prior to stroke. Genetic pathways unique to diet-induced obesity were primarily related to adaptive immunity, extracellular matrix components, cell migration, and vasculogenesis. The late appearance of neuroinflammation and infarct expansion indicates that there may be a therapeutic window between 10 and 36 h after stroke where inflammation and obesity-specific transcriptional programs could be targeted to improve outcomes in people with obesity and stroke.


Patients' perceptions of interpersonal continuity of care.
The health system shift from doctor-patient continuity of care to team-based continuity may not match patients' preferences. This article reviews the existing medical literature regarding patients' perceptions of interpersonal continuity of care to determine which patients value interpersonal continuity and in what context. A search of the ISI Web of Knowledge database from 1970 to April 2005 and the MEDLINE database from 1966 through April 2005 was performed to find articles that elicited patients' preferences or value for continuity of care. Thirty six articles were used for this synthesis. Interpersonal continuity of care is important to a majority of patients, particularly those from vulnerable groups. Patients value the relationship with their physician, their physician's knowledge about them, and the ability to communicate their concerns. Over time, contact with a physician seems to lead to the development of trust and confidence. However, continuity of care is not valued by all patients or across all settings. Future research should continue to explore the importance of interpersonal continuity to patients. In particular, are there medical consequences when those whom desire continuity do not receive it? Clinicians should consider incorporating patients' preference for continuity into their office scheduling procedures.


A prospective study of folate intake and the risk of breast cancer.
Folate is involved in DNA synthesis and methylation and may reduce breast cancer risk, particularly among women with greater alcohol consumption. To assess the association between folate intake and risk of breast cancer and whether higher folate intake may reduce excess risk among women who consume alcohol. Prospective cohort study performed in 1980, with 16 years of follow-up. A total of 88818 women who completed the dietary questionnaire section of the Nurses' Health Study in 1980. Incidence of invasive breast cancer by levels of folate and alcohol intake. A total of 3483 cases of breast cancer were documented. Total folate intake was not associated with overall risk of breast cancer. However, among women who consumed at least 15 g/d of alcohol, the risk of breast cancer was highest among those with low folate intake. For total folate intake of at least 600 microg/d compared with 150 to 299 microg/d, the multivariate relative risk (RR) was 0.55 (95% confidence interval [CI], 0.39-0.76; P for trend = .001). This association was only slightly attenuated after additional adjustment for intake of beta carotene, lutein/zeaxanthin, preformed vitamin A, and total vitamins C and E. The risk of breast cancer associated with alcohol intake was strongest among women with total folate intake of less than 300 microg/d (for alcohol intake > or =15 g/d vs <15 g/d, multivariate RR, 1.32; 95% CI, 1.15-1.50). For women who consumed at least 300 microg/d of total folate, the multivariate RR for intake of at least 15 g/d of alcohol vs less than 15 g/d was 1.05 (95% CI, 0.92-1.20). Current use of multivitamin supplements, the major source of folate, was associated with lower breast cancer risk among women who consumed at least 15 g/d of alcohol (for current users of supplements vs never users, RR, 0.74; 95% CI, 0.59-0.93). Our findings suggest that the excess risk of breast cancer associated with alcohol consumption may be reduced by adequate folate intake.


In search of optimal fear inducing stimuli: Differential behavioral responses to computer animated images in zebrafish.
Zebrafish has been gaining popularity in behavioral genetics and behavioral neuroscience as this species offers an excellent compromise between system complexity and practical simplicity for mechanistic analyses of brain and behavior function. Recently, a number of studies started to investigate methods with which fear responses may be induced reliably in zebrafish. The ultimate goal of these studies has been to develop zebrafish models of pathological processes and to investigate the mechanisms of fear and to eventually translate the findings to the human clinic. Previously, animated image of a sympatric predator of zebrafish was shown to induce fear responses. Here we expand on this recently gained knowledge and investigate whether other moving images may induce more robust fear responses. The images investigated include the original sympatric predator, the Indian leaf fish, another sympatric predator, the needle fish, a bird silhouette moved on the side or above the tank, an expanding dot mimicking rapid approach of an object shown on the side and from above the tank, as well as non-fear inducing images including a single and a group of zebrafish. Our results indicate that although the sympatric predators do induce some fear responses, the other images, particularly the expanding dot but also the bird silhouette shown from above are more effective. The results also reveal a stimulus dependent motor pattern response repertoire of zebrafish demonstrating that perhaps univariate quantification methods may not be appropriate for uncovering the complexity of fear or anxiety related phenotypical changes in this species.


Concordance of Demographic Characteristics, Sexual Behaviors, and Relationship Attributes Among Sex Dyads of Black and White Men Who Have Sex with Men.
Differences in individual behaviors have failed to explain racial disparities between Black and White men who have sex with men (MSM). However, reporting of behaviors and partner characteristics are assumed to be non-differentially reported by race. From 314 participants, this study used the two-sided data-where sexual partners provide information on each other and their relationship-of 127 dyads of Black and White MSM from Atlanta, GA, to assess the reliability of partner-reported demographic characteristics and the concordance of sexual behaviors and partnership attributes by race. We compared proportions of concordance by race using a modified kappa (K m) to assess chance-corrected agreement. The median difference in age between self- and partner-reports was 0 (0-1) years. Compared to self-reports, 97 % of the partners of Black participants and 96 % of the partners of White participants correctly classified their race. We observed poor agreement on pre-sexual discussion (K m = 0.18) and being in an ongoing relationship (K m = 0.13), with no differences by race (p = 0.11). Although not statistically significant, Black MSM dyads had lower levels of concordance for unprotected anal intercourse in the previous 12 months (68 %) compared to White dyads (90 %), with fair agreement among Black dyads (K m = 0.26). Measures of partner-reported age and race are likely accurate; however, certain self-reported sexual behaviors and partnership attributes may be unreliable and differentially reported by race. Our findings highlight the need to assess the validity of measures used to estimate HIV transmission and inform racial disparities research.


Effects of in utero arsenic exposure on child immunity and morbidity in rural Bangladesh.
Chronic exposure to arsenic, a potent carcinogen and toxicant, via drinking water is a worldwide public health problem. Because little is known about early-life effects of arsenic on immunity, we evaluated the impact of in utero exposure on infant immune parameters and morbidity in a pilot study. Pregnant women were enrolled at 6-10 weeks of gestation in Matlab, a rural area of Bangladesh, extensively affected by arsenic contamination of tubewell water. Women (n=140) delivering at local clinics were included in the study. Anthropometry and morbidity data of the pregnant women and their children, as well as infant thymic size by sonography were collected. Maternal urine and breast milk were collected for immune marker and arsenic assessment. Maternal urinary arsenic during pregnancy showed significant negative correlation with interleukin-7 (IL-7) and lactoferrin (Ltf) in breast milk and child thymic index (TI). Urinary arsenic was also positively associated with fever and diarrhea during pregnancy and acute respiratory infections (ARI) in the infants. The effect of arsenic exposure on ARI was only evident in male children. The findings suggest that in utero arsenic exposure impaired child thymic development and enhanced morbidity, probably via immunosuppression. The effect seemed to be partially gender dependent. Arsenic exposure also affected breast milk content of trophic factors and maternal morbidity.


NATpare: a pipeline for high-throughput prediction and functional analysis of nat-siRNAs.
Natural antisense transcript-derived small interfering RNAs (nat-siRNAs) are a class of functional small RNA (sRNA) that have been found in both plant and animals kingdoms. In plants, these sRNAs have been shown to suppress the translation of messenger RNAs (mRNAs) by directing the RNA-induced silencing complex (RISC) to their sequence-specific mRNA target(s). Current computational tools for classification of nat-siRNAs are limited in number and can be computationally infeasible to use. In addition, current methods do not provide any indication of the function of the predicted nat-siRNAs. Here, we present a new software pipeline, called NATpare, for prediction and functional analysis of nat-siRNAs using sRNA and degradome sequencing data. Based on our benchmarking in multiple plant species, NATpare substantially reduces the time required to perform prediction with minimal resource requirements allowing for comprehensive analysis of nat-siRNAs in larger and more complex organisms for the first time. We then exemplify the use of NATpare by identifying tissue and stress specific nat-siRNAs in multiple Arabidopsis thaliana datasets.


The beta agonist lung injury trial prevention. A randomized controlled trial.
Experimental studies suggest that pretreatment with β-agonists might prevent acute lung injury (ALI). To determine if in adult patients undergoing elective esophagectomy, perioperative treatment with inhaled β-agonists effects the development of early ALI. We conducted a randomized placebo-controlled trial in 12 UK centers (2008-2011). Adult patients undergoing elective esophagectomy were allocated to prerandomized, sequentially numbered treatment packs containing inhaled salmeterol (100 μg twice daily) or a matching placebo. Patients, clinicians, and researchers were masked to treatment allocation. The primary outcome was development of ALI within 72 hours of surgery. Secondary outcomes were ALI within 28 days, organ failure, adverse events, survival, and health-related quality of life. An exploratory substudy measured biomarkers of alveolar-capillary inflammation and injury. A total of 179 patients were randomized to salmeterol and 183 to placebo. Baseline characteristics were similar. Treatment with salmeterol did not prevent early lung injury (32 [19.2%] of 168 vs. 27 [16.0%] of 170; odds ratio [OR], 1.25; 95% confidence interval [CI], 0.71-2.22). There was no difference in organ failure, survival, or health-related quality of life. Adverse events were less frequent in the salmeterol group (55 vs. 70; OR, 0.63; 95% CI, 0.39-0.99), predominantly because of a lower number of pneumonia (7 vs. 17; OR, 0.39; 95% CI, 0.16-0.96). Salmeterol reduced some biomarkers of alveolar inflammation and epithelial injury. Perioperative treatment with inhaled salmeterol was well tolerated but did not prevent ALI. Clinical trial registered with International Standard Randomized Controlled Trial Register (ISRCTN47481946) and European Union database of randomized Controlled Trials (EudraCT 2007-004096-19).


Cannabis use as a risk factor for causing motor vehicle crashes: a prospective study.
We conducted a responsibility analysis to determine whether drivers injured in motor vehicle collisions who test positive for Δ-9-tetrahydrocannabinol (THC) or other drugs are more likely to have contributed to the crash than those who test negative. Prospective case-control study. Trauma centres in British Columbia, Canada. Injured drivers who required blood tests for clinical purposes following a motor vehicle collision. Excess whole blood remaining after clinical use was obtained and broad-spectrum toxicology testing performed. The analysis quantified alcohol and THC and gave semiquantitative levels of other impairing drugs and medications. Police crash reports were analysed to determine which drivers contributed to the crash (responsible) and which were 'innocently involved' (non-responsible). We used unconditional logistic regression to determine the likelihood (odds ratio: OR) of crash responsibility in drivers with 0 < THC < 2 ng/ml, 2 ng/ml ≤ THC < 5 ng/ml and THC ≥ 5 ng/ml (all versus THC = 0 ng/ml). Risk estimates were adjusted for age, sex and presence of other impairing substances. We obtained toxicology results on 3005 injured drivers and police reports on 2318. Alcohol was detected in 14.4% of drivers, THC in 8.3%, other drugs in 8.9% and sedating medications in 19.8%. There was no increased risk of crash responsibility in drivers with THC < 2 ng/ml or 2 ≤ THC < 5 ng/ml. In drivers with THC ≥ 5 ng/ml, the adjusted OR was 1.74 [95% confidence interval (CI) = 0.59-6.36; P = 0.35]. There was significantly increased risk of crash responsibility in drivers with blood alcohol concentration (BAC) ≥ 0.08% (OR = 6.00;95% CI = 3.87-9.75; P < 0.01), other recreational drugs detected (OR = 1.82;95% CI = 1.21-2.80; P < 0.01) or sedating medications detected (OR = 1.45; 95%CI = 1.11-1.91; P < 0.01). In this sample of non-fatally injured motor vehicle drivers in British Columbia, Canada, there was no evidence of increased crash risk in drivers with Δ-9-tetrahydrocannabinol < 5 ng/ml and a statistically non-significant increased risk of crash responsibility (odds ratio = 1.74) in drivers with Δ-9-tetrahydrocannabinol ≥ 5 ng/ml.


Rehabilitation after successful finger replantation.
The aim of the present study was to assess results of rehabilitation of patients after finger replantation. The study examined 160 fingers amputated and replanted at various levels between 2000 and 2013 at the clinic. Mean patient age was 29.4 years. Mean follow-up time was 23 months. Rehabilitation of fingers began between postoperative fourth and eighth week and continued until the 24th week. Range of motion of affected hand, return to daily activities, aesthetic appearance, and patient satisfaction were assessed according to Tamai criteria. Functional results according to Tamai criteria were perfect in 36 patients, good in 54 patients, average in 27 patients, and poor in 18 patients. Post-operative rehabilitation of replanted fingers should begin as soon as possible. During the rehabilitation period, physiotherapist, surgeon, and patient must work in close cooperation. Functional results of patients who adjust to the rehabilitation program, home practice, and splint usage are better.


Exclusion from Social Relations in Later Life: Micro- and Macro-Level Patterns and Correlations in a European Perspective.
Older adults face particular risks of exclusion from social relationships (ESR) and are especially vulnerable to its consequences. However, research so far has been limited to specific dimensions, countries, and time points. In this paper, we examine the prevalence and micro- and macro-level predictors of ESR among older adults (60+) using two waves of data obtained four years apart across 14 European countries in the Survey of Health, Ageing and Retirement in Europe (SHARE). We consider four ESR indicators (household composition, social networks, social opportunities, and loneliness) and link them to micro-level (age, gender, socioeconomic factors, health, and family responsibilities) and national macro-level factors (social expenditures, unmet health needs, individualism, social trust, and institutional trust). Findings reveal a northwest to southeast gradient, with the lowest rates of ESR in the stronger welfare states of Northwest Europe. The high rates of ESR in the southeast are especially pronounced among women. Predictably, higher age and fewer personal resources (socioeconomic factors and health) increase the risk of all ESR dimensions for both genders. Macro-level factors show significant associations with ESR beyond the effect of micro-level factors, suggesting that national policies and cultural and structural characteristics may play a role in fostering sociability and connectivity and, thus, reduce the risk of ESR in later life.