GATA1 downregulation in prefibrotic and fibrotic stages of primary myelofibrosis and in the myelofibrotic progression of other myeloproliferative neoplasms.
GATA binding protein 1 (GATA1) is a transcription factor essential for effective erythropoiesis and megakaryopoiesis. Two isoforms of GATA1 exist, derived from alternative splicing. "GATA1" is the full length and functionally active protein; "GATA1s" is the truncated isoform devoid of the activation domain, the function of which has not been fully elucidated. Reduced megakaryocytic expression of GATA1 has been linked to impaired hematopoiesis and bone marrow fibrosis in murine models and in vivo in patients affected by primary myelofibrosis (PMF). However, data is limited regarding GATA1 expression in other myeloproliferative neoplasms (MPN) such as pre-fibrotic PMF (pre-PMF), polycythemia vera (PV) and essential thrombocythemia (ET) and in their respective fibrotic progression. To assess whether an immunohistologic approach can be of help in separating different MPN, we have performed a comprehensive immunohistochemical evaluation of GATA1 expression in megakaryocytes within a cohort of BCR-ABL1 negative MPN. In order to highlight any potential differences between the two isoforms we tested two clones, one staining the sum of GATA1 and GATA1s ("clone 1"), the other staining GATA1 full length alone ("clone 2"). At the chronic phase, a significant reduction preferentially of GATA1 full length was seen in pre-fibrotic PMF, particularly compared to ET and PV; no significant differences were observed between PV and ET. The fibrotic progression of both PV and ET was associated with a significant reduction in GATA1, particularly affecting the GATA1 full length isoform. The fibrotic progression of pre-PMF to PMF was associated with a significant reduction of the overall GATA1 protein and a trend in reduction of GATA1s. Our findings support a role of GATA1 in the pathogenesis of BCR-ABL1 negative MPN, particularly in their fibrotic progression and suggest that the immunohistochemical evaluation of GATA1 may be of use in the differential diagnosis of these neoplasms.


Is the DHEAS/cortisol ratio a potential filter for non-operable constipated cases?
Constipation is a significant manifestation of a number of psychological disorders. Published papers recommend using self-assessment questionnaires for discriminating psychological from non-psychological constipated patients before operating on them but reports from major surveys revealed that general practitioners failed to diagnose 70% of depressed patients using self-assessment questionnaires. Lower circulating concentrations of progesterone, 17-hydroxyprogesterone, cortisol, testosterone, androstenedione, and dehydroepiandrostenedione sulfate (DHEAS) during the follicular phase in constipated young women compared with respective controls were found during the follicular phase of the menstrual cycles. During the luteal phase of the cycle, reductions were identified in estriol, cortisol and testosterone in the constipated group. Likewise, circulating concentrations of DHEAS were found to be lower in depressed patients than comparable healthy controls. DHEAS/cortisol ratios in morning serum and salivary samples were lower than those retrieved during other times of the day in depressed patients. The idea of recognizing major depression in constipated patients by measuring DHEAS/cortisol ratios in saliva and serum may be plausible but this possibility needs to be confirmed in well-designed studies.


Remote Postischemic Conditioning Promotes Stroke Recovery by Shifting Circulating Monocytes to CCR2+ Proinflammatory Subset.
Brain injury from stroke is typically considered an event exclusive to the CNS, but injury progression and repair processes are profoundly influenced by peripheral immunity. Stroke stimulates an acute inflammatory response that results in a massive infiltration of peripheral immune cells into the ischemic area. While these cells contribute to the development of brain injury, their recruitment has been considered as a key step for tissue repair. The paradoxical role of inflammatory monocytes in stroke raises the possibility that the manipulation of peripheral immune cells before infiltration into the brain could influence stroke outcome. One such manipulation is remote ischemic limb conditioning (RLC), which triggers an endogenous tolerance mechanism. We observed that mice subjected to poststroke RLC shifted circulating monocytes to a CCR2+ proinflammatory monocyte subset and had reduced acute brain injury, swelling, and improved motor/gait function in chronic stroke. The RLC benefits were observed regardless of injury severity, with a greater shift to a CCR2+ subset in severe stroke. Adoptive transfer of CCR2-deficient monocytes abolished RLC-mediated protection. The study demonstrates the importance of RLC-induced shift of monocytes to a CCR2+ proinflammatory subset in attenuating acute injury and promoting functional recovery in chronic stroke. The defined immune-mediated mechanism underlying RLC benefits allows for an evidence-based framework for the development of immune-based therapeutic strategies for stroke patients.SIGNIFICANCE STATEMENT Stroke is the leading cause of physical disability worldwide but has few treatment options for patients. Because remote ischemic limb conditioning (RLC) elicits endogenous tolerance in neither an organ- nor a tissue-specific manner, the immune system has been considered a mediator for an RLC-related benefit. Application of RLC after stroke increased a proinflammatory CCR2+ monocyte subset in the blood and the brain. RLC reduced acute stroke injury and promoted motor/gait function during the recovery phase. The RLC benefits were absent in mice that received CCR2-deficient monocytes. This preclinical study shows the importance of CCR2+ proinflammatory monocytes in RLC benefits in stroke and provides a therapeutic RLC platform as a novel immune strategy to improve outcomes in stroke patients.


Purification and characterization of a novel β-glucosidase from Aspergillus flavus and its application in saccharification of soybean meal.
Aspergillus flavus has been regarded as a potential candidate for its production of industrial enzymes, but the details of β-glucosidase from this strain is very limited. In herein, we first reported a novel β-glucosidase (AfBglA) with the molecular mass of 94.2 kDa from A. flavus. AfBglA was optimally active at pH 4.5 and 60 °C and is stable between pH 3.5 and 9.0 and at a temperature of up to 55 °C for 30 min remaining more than 90% of its initial activity. It showed an excellent tolerance to Trypsin, Pepsin, Compound Protease, and Flavourzyme and its activity was not inhibited by specific certain cations. AfBglA displayed broad substrate specificity, it acted on all tested pNP-glycosides and barley glucan, indicating this novel β-glucosidase exhibited a β-1, 3-1, 4-glucanase activity. Moreover, the AfBglA could effectively hydrolyze the soybean meal suspension into glucose and exhibit a strong tolerance to the inhibition of glucose at a concentration of 20.0 g/L during the saccharification. The maximum amount of the glucose obtained by AfBglA corresponded to 67.0 g/kg soybean meal. All of these properties mentioned above indicated that the AfBglA possibly attractive for food and feed industry and saccharification of cellulolytic materials.


Next-Generation Sequencing Approaches in Cancer: Where Have They Brought Us and Where Will They Take Us?
Next-generation sequencing (NGS) technologies and data have revolutionized cancer research and are increasingly being deployed to guide clinicians in treatment decision-making. NGS technologies have allowed us to take an "omics" approach to cancer in order to reveal genomic, transcriptomic, and epigenomic landscapes of individual malignancies. Integrative multi-platform analyses are increasingly used in large-scale projects that aim to fully characterize individual tumours as well as general cancer types and subtypes. In this review, we examine how NGS technologies in particular have contributed to "omics" approaches in cancer research, allowing for large-scale integrative analyses that consider hundreds of tumour samples. These types of studies have provided us with an unprecedented wealth of information, providing the background knowledge needed to make small-scale (including "N of 1") studies informative and relevant. We also take a look at emerging opportunities provided by NGS and state-of-the-art third-generation sequencing technologies, particularly in the context of translational research. Cancer research and care are currently poised to experience significant progress catalyzed by accessible sequencing technologies that will benefit both clinical- and research-based efforts.


Early changes in bone density, microarchitecture, bone resorption, and inflammation predict the clinical outcome 12 weeks after conservatively treated distal radius fractures: an exploratory study.
Fracture healing is an active process with early changes in bone and inflammation. We performed an exploratory study evaluating the association between early changes in densitometric, structural, biomechanical, and biochemical bone parameters during the first weeks of fracture healing and wrist-specific pain and disability at 12 weeks in postmenopausal women with a conservatively treated distal radius fracture. Eighteen patients (aged 64 ± 8 years) were evaluated at 1 to 2 and 3 to 4 weeks postfracture, using high-resolution peripheral quantitative computed tomography (HR-pQCT), micro-finite element analysis, serum procollagen type-I N-terminal propeptide (P1NP), carboxy-terminal telopeptide of type I collagen (ICTP), and high-sensitive C-reactive protein (hsCRP). After 12 weeks, patients rated their pain and disability using Patient Rated Wrist Evaluation (PRWE) questionnaire. Additionally, Quick Disability of the Arm Shoulder and Hand (QuickDASH) questionnaire and active wrist range of motion was evaluated. Linear regression models were used to study the relationship between changes in bone parameters and in hsCRP from visit 1 to 2 and PRWE score after 12 weeks. A lower PRWE outcome, indicating better outcome, was significantly related to an early increase in trabecular bone mineral density (BMD) (β -0.96 [95% CI -1.75 to -0.16], R(2) = 0.37), in torsional stiffness (-0.14 [-0.28 to -0.004], R(2) = 0.31), and to an early decrease in trabecular separation (209 [15 to 402], R(2) = 0.33) and in ICTP (12.1 [0.0 to 24.1], R(2) = 0.34). Similar results were found for QuickDASH. Higher total dorsal and palmar flexion range of motion was significantly related to early increase in hsCRP (9.62 [3.90 to 15.34], R(2) = 0.52). This exploratory study indicates that the assessment of early changes in trabecular BMD, trabecular separation, calculated torsional stiffness, bone resorption marker ICTP, and hsCRP after a distal radius fracture provides valuable information regarding the 12-week clinical outcome in terms of pain, disability, and range of motion and validates its use in studies on the process of early fracture healing. © 2014 American Society for Bone and Mineral Research.


Differences in replication of a DNA template containing an ethyl phosphotriester by T4 DNA polymerase and Escherichia coli DNA polymerase I.
A DNA template containing a single ethyl phosphotriester was replicated in vitro by the bacteriophage T4 DNA polymerase and by Escherichia coli DNA polymerase I (DNA pol I). Escherichia coli DNA pol I bypassed the lesion efficiently, but partial inhibition was observed for T4 DNA polymerase. The replication block produced by the ethyl phosphotriester was increased at low dNTP concentrations and for a mutant T4 DNA polymerase with an antimutator phenotype, increased proofreading activity, and reduced ability to bind DNA in the polymerase active center. These observations support a model in which an ethyl phosphotriester impedes primer elongation by T4 DNA polymerase by decreasing formation of the ternary DNA polymerase-DNA-dNTP complex. When primer elongation is not possible, proofreading becomes the favored reaction. Apparent futile cycles of nucleotide incorporation and proofreading, the idling reaction, were observed at the site of the lesion. The replication block was overcome by higher dNTP concentrations. Thus, ethyl phosphotriesters may be tolerated in vivo by the up-regulation of dNTP biosynthesis that occurs during the cellular checkpoint response to blocked DNA replication forks.


Usefulness of the Calgary Syncope Symptom Score for the diagnosis of vasovagal syncope in the elderly.
The Calgary Syncope Symptom Score (CSSS) has been validated as a simple point score of historical features with high sensitivity and specificity for the diagnosis of vasovagal syncope (VVS) in younger populations without evidence of structural heart disease. Our purpose was to evaluate the performance of the CSSS in an elderly population with suspected VVS. Hundred and eighty patients of ≥60 years of age (mean 73.4 ± 7.8) with suspected clinical diagnosis of VVS were studied. The CSSS (VVS score ≥-2) was calculated in all patients prior to undergoing head-up tilt test (HUT). A standardized HUT protocol with active nitroglycerin phase was used to reproduce syncopal symptoms as gold standard for diagnosis of VVS. Hundred and forty patients had positive HUT response. Eighty-three patients (42.3%) had CSSS ≥-2 suggesting a diagnosis of VVS. The Calgary Syncope Symptom Score sensitivity was 0.51 [95% confidence interval (CI) 0.42-0.59] and specificity 0.73 (95% CI 0.52-0.85) with positive predictive value and negative predictive value of 0.87 (95% CI 0.77-0.93) and 0.30 (95% CI 0.21-0.40), respectively. One hundred (55.6%) patients had previous history of mild cardiovascular disease documented during assessment prior to HUT. In this population sensitivity and specificity was markedly reduced: 0.13 (95% CI 0.05-0.29) and 0.70 (95% CI 0.57-0.80), respectively. The CSSS has a lower sensitivity and specificity in an elderly population presenting with syncope compared to previously validated data in young adults, particularly in elderly patients with previous history of mild cardiovascular disease. A modified CSSS may be needed to improve specificity and sensitivity in this population.


Saphenous vein bypass to pedal arteries. An aggressive strategy for foot salvage.
Patients with forefoot ischemia and severe tibial artery disease present a major challenge to revascularization and foot preservation. Encouraged by the success of saphenous vein bypass to the more proximal arteries of the lower leg, we extended this technique to the pedal arteries. Between February 1986 and September 1988, we performed 26 bypasses in 24 men (mean age, 66 years) with critical foot ischemia. Sixty-three percent of the patients had diabetes mellitus, 50% had hypertension, and 71% were actively smoking. Angiography invariably revealed multiple tibial artery occlusions with reconstitution of the pedal arteries. The foot salvage rate was 83% (mean survival, 14 months), primary patency was 83% (mean survival, 9 months), and survival was 86% (mean, 12 months). There were 11 wound complications (42%); two resulted in disruptions of the distal anastomosis and eventual graft failure. Bypass to the pedal arteries yields a high rate of foot salvage and is comparable with more proximal bypass procedures; however, wound complications are common and require special technical considerations.


Effects of oral α-lipoic acid administration on body weight in overweight or obese subjects: a crossover randomized, double-blind, placebo-controlled trial.
Alpha-lipoic acid (ALA) has shown beneficial properties on diabetes and obesity. The aim of this study was to examine the effects of oral ALA on body weight in subjects with overweight or obese. Single-centre, randomized, double-blind, crossover controlled study. A total of 166 subjects of Chinese Han ethnicity with a BMI ≥25 kg/m2 were screened and 103 subjects fulfilled the study requirements, in terms of informed consent and participation to the study. The subjects were randomized (1:1) to receive either ALA (1200 mg/day) or placebo treatment in a crossover design for 8 weeks. The primary end-point was the change in body weight. The secondary end-points were the changes in waist circumference, BMI, lipid profile, plasma leptin levels and the adverse events that occurred following ALA treatment. The changes in the body weight and waist circumference noted in the ALA group were significantly different compared to the placebo group as demonstrated by mixed model statistical analysis (both P < 0·05). No real weight reduction was seen in the ALA group, and no significant differences were noted as regards cholesterol levels, triglyceride levels, high-density lipoprotein cholesterol levels and adverse events between the two groups. The administration of ALA was well tolerated, and no serious adverse events were noted. Oral administration of ALA (1200 mg/day) for 8 weeks induced mild weight loss accompanied by a reduction in waist circumference.


The effects of oral contraceptives on androgen levels and their relevance to premenstrual mood and sexual interest: a comparison of two triphasic formulations containing norgestimate and either 35 or 25 microg of ethinyl estradiol.
This study compared two oral contraceptives (OCs) with the same triphasic regimen of progestin (norgestimate 0.18, 0.215 and 0.25 mg) but differing doses of ethinyl estradiol (EE) - 25 and 35 microg EE - in their effects on androgens, mood and sexual interest in women starting on OCs. Total testosterone (T), free testosterone (FT), sex-hormone-binding globulin (SHBG) and dehydroepiandrosterone sulphate (DHEA-S), together with measures of mood [Beck Depression Inventory (BDI)], sexual interest [Dyadic and Solitary subscales of the Sexual Desire Inventory (SDI)] and self-reported side effects were assessed before starting on the OC and again after 3 months of use. Sixty women, all university students, were randomized to receive either the 25 microg EE (N/EE25) or the 35 microg EE (N/EE35) pill; 12 women discontinued, leaving 48 who completed the 3-month study. Their mean age was 19.7 years (18-30) and they were predominantly white and single. Both OCs produced reductions in mean T [N/EE35: from 1.33 to 0.60 nmol/L, p<.001; N/EE25: from 1.12 to 1.02 nmol/L; nonsignificant (NS)] and FT (N/EE35: from 41.3 to 4.4 pmol/L, p<.001; N/EE25: from 25.4 to 7.9 pmol/L, p<.01), but the reduction in both T and FT was significantly greater with the higher EE dose (N/EE35) (p=.05 and p=.03, respectively). DHEA-S was also reduced with both formulations (N/EE35: from 7.26 to 5.22 micromol/L); N/EE25: from 7.50 to 5.39 micromol/L), although the reduction was only significant in the N/EE35 group (p<.02). Considerable variability in changes in mood was evident with both OCs, with some women showing predominantly negative effects (10 in N/EE35, 5 in N/EE25); others, positive effects (9 in N/EE35, 17 in N/EE25) and some, no change (four in each group). Women using N/EE25 were significantly more likely to show improvement in premenstrual mood than those in the N/EE35 group (p<.02), although there was no correlation between changes in BDI and FT or DHEA-S. Sexual interest scores did not change significantly from baseline to posttreatment with either OC (N/EE35: dyadic, from 40.5 to 39.6, NS; solitary, from 5.9 to 6.4, NS; N/EE25: dyadic, from 36.7 to 37.0, NS; solitary, from 5.0 to 4.2, NS). The lower EE pill reduced FT less and was associated with greater improvement in premenstrual mood. A causal relation between these two effects is uncertain.


Phylogenetic and functional analyses of the cytochrome P450 family 4.
Cytochrome P450 family 4 (CYP4) proteins metabolize fatty acids, eicosanoids, and vitamin D and are important for chemical defense. The purpose of this study was to determine the evolutionary relationships between vertebrate CYP4 subfamilies and raise functional hypotheses regarding CYP4 subfamilies with little empirical data. 132 CYP4 sequences from 28 species were utilized for phylogenetic reconstructions by maximum likelihood and Bayesian inference. Monophyly was not found with the CYP4T and CYP4B subfamilies. CYP4V clustered with invertebrate subfamilies. Evolutionary rates of functional divergence were high in pairwise comparison with CYP4X yet, comparisons with mammalian CYP4F22 genes generally had no statistically significant divergence. Radical biochemical changes were detected in regions associated with substrate binding and the active site in comparisons among the CYP4A, CYP4X, and CYP4B subfamilies. Lastly, gene expression patterns, determined in silico with EST libraries from human, chicken, frog and fish, for CYP4V was markedly different between human and actinopterygian species. Further consideration should be given to the nomenclature of the CYP4T and CYP4B subfamily genes. Strong support was seen for the placement of CYP4A as a basal subfamily to CYP4X and CYP4Z. The B, B', J', K', K″ helices and a region at the end of C-terminus were suggested as conserved regions in CYP4 genes. The function of CYP4X was hypothesized to specialize in metabolism of long chain fatty acids. CYP4F22 genes may share a similar function to other CYP4F genes, although gene expression sites were different.


Role of Bi-Atrial Pacing In Slowing The Progression of Paroxysmal Atrial Fibrillation To Permanent Atrial Fibrillation.
Introduction: Bi-atrial lead placement combined with atrial overdrive pacing has demonstrated a reduction in percent time mode switched and mode switches per day. This retrospective analysis compared long term outcomes of patients with right atrial overdrive pacing alone (DAO) to patients having atrial overdrive with bi-atrial leads (BIA) in slowing the progression of paroxysmal atrial fibrillation (PAF) to permanent continuous atrial fibrillation (CAF). Methods: Thirty-three patients age 76.6 (+/-1.96) from our prior investigation were selected. The DAO control group (N=16) had received a standard right atrial pacing lead. The BIA group (N=17) had pacing leads placed in the right atrium and coronary sinus. Patients were followed for a mean 1217 days (+/-838). Days of CAF was classified as the date of final mode switch until analysis. Results: A total of 40,171 follow-up days were evaluated. The mean follow-up for both cohorts was 1217 days (+/-838). The DAO group consisted of 15,318 days (mean 957 +/-761) and the BIA group 24,853 days (mean 1461 +/-854). A lower total number of days were spent in CAF in the BIA group versus the DAO group, 1380 vs 2197 respectively. Corrected for follow-up duration, 5.55% days in CAF was seen in the BIA group vs. 14.34% in the DAO group which did not reach statistical significance. Conclusions: Although BIA overdrive pacing initially demonstrated reduced time in mode switch compared to DAO alone, this analysis did not detect a reduction in progression to CAF. More subjects or a longer follow up would be needed.


Perspectives on reasons of medication nonadherence in psychiatric patients.
This study was carried out to evaluate factors resulting in medication nonadherence within 6 months before admission to the psychiatric service of our hospital for bipolar disorder, schizophrenia/schizoaffective disorder, depression, and other psychiatric diseases. Two hundred and three patients admitted to the Psychiatry Service of the Medical Faculty were included in this study. Sociodemographic parameters and clinical findings within 6 months before admission and patients' views on reasons of medication nonadherence were examined. Patients were classified into four groups according to their diagnosis: bipolar disorder (n=68, 33.5%), schizophrenia/schizoaffective disorder (n=59, 29.1%), depression (n=39, 19.2%), and others (n=37, 18.2%). The ratio of medication nonadherence was higher in the bipolar disorder group when compared to the groups with schizophrenia/schizoaffective disorder, depression, and other disorders (12.1%, 18.2%, and 24.2% vs 45.5%); however, the ratio of medication nonadherence was similar in schizophrenia/schizoaffective disorder, depression, and the others group. In logistic regression analysis, irregular follow-up (odds ratio [OR]: 5.7; 95% confidence interval [CI]: 2.92-11.31) and diagnosis (OR: 1.5; 95% CI: 1.07-1.95) were determined to be important risk factors for medication nonadherence. The leading factors for medication nonadherence were: "not willing to use medication", "not accepting the disease", and "being disturbed by side effects" in the bipolar disorder group, "not accepting the disease" in the schizophrenia/schizoaffective disorder group, "feeling well" in the depression group, and "being disturbed by side effects" in the other diseases group. Medication nonadherence is an important problem in psychiatric patients and should be dealt with by taking into account the diagnosis, attendance to follow-up appointments, and the patient's attitude. Ensuring regular attendance to follow-up appointments, adjusting the management plan according to the diagnosis, and improving their thoughts about resistance to medication can be beneficial in terms of medication adherence.


Attitudes towards practice nurses--survey of a sample of general practitioners in England and Wales.
Practice nursing has expanded rapidly since the 1990 contract for general practitioners. In 1990, a national survey was undertaken of the attitudes of a random sample of general practitioners towards practice nurses. Responses to the postal questionnaire were received from 41.9% of the 4800 general practitioners sampled. Of the responding general practitioners, 90.0% were satisfied with the role of the practice nurse within their practice. To fulfil the requirements of the 1990 contract for general practitioners 50.7% had created a new nursing post, and 83.1% had expanded the role of nurses already employed; 89.7% wished to see further expansion of the practice nurse's role. However, lack of space was the factor most frequently reported as limiting the expansion of the practice nurse's role, mentioned by 76.0% of general practitioners. Only 43.7% of general practitioners recognized lack of opportunities for practice nurse training as a hindrance to role expansion. The key to managing the expansion of the role of the practice nurse lies in the provision of resources and in training. A pressing need exists for a national training scheme based in general practice.


An FBXW7-ZEB2 axis links EMT and tumour microenvironment to promote colorectal cancer stem cells and chemoresistance.
Colorectal cancer (CRC) patients develop recurrence after chemotherapy owing to the survival of stem cell-like cells referred to as cancer stem-like cells (CSCs). The origin of CSCs is linked to the epithelial-mesenchymal transition (EMT) process. Currently, it remains poorly understood how EMT programmes enable CSCs residing in the tumour microenvironment to escape the effects of chemotherapy. This study identifies a key molecular pathway that is responsible for the formation of drug-resistant CSC populations. Using a modified yeast-2-hybrid system and 2D gel-based proteomics methods, we show that the E3-ubiquitin ligase FBXW7 directly binds and degrades the EMT-inducing transcription factor ZEB2 in a phosphorylation-dependent manner. Loss of FBXW7 induces an EMT that can be effectively reversed by knockdown of ZEB2. The FBXW7-ZEB2 axis regulates such important cancer cell features, as stemness/dedifferentiation, chemoresistance and cell migration in vitro, ex vivo and in animal models of metastasis. High expression of ZEB2 in cancer tissues defines the reduced ZEB2 expression in the cancer-associated stroma in patients and in murine intestinal organoids, demonstrating a tumour-stromal crosstalk that modulates a niche and EMT activation. Our study thus uncovers a new molecular mechanism, by which the CRC cells display differences in resistance to chemotherapy and metastatic potential.


The molecular mechanism underlying the proliferating and preconditioning effect of vitamin C on adipose-derived stem cells.
Although adipose-derived stem cells (ASCs) show promise for cell therapy, there is a tremendous need for developing ASC activators. In the present study, we investigated whether or not vitamin C increases the survival, proliferation, and hair-regenerative potential of ASCs. In addition, we tried to find the molecular mechanisms underlying the vitamin C-mediated stimulation of ASCs. Sodium-dependent vitamin C transporter 2 (SVCT2) is expressed in ASCs, and mediates uptake of vitamin C into ASCs. Vitamin C increased the survival and proliferation of ASCs in a dose-dependent manner. Vitamin C increased ERK1/2 phosphorylation, and inhibition of the mitogen-activated protein kinase (MAPK) pathway attenuated the proliferation of ASCs. Microarray and quantitative polymerase chain reaction showed that vitamin C primarily upregulated expression of proliferation-related genes, including Fos, E2F2, Ier2, Mybl1, Cdc45, JunB, FosB, and Cdca5, whereas Fos knock-down using siRNA significantly decreased vitamin C-mediated ASC proliferation. In addition, vitamin C-treated ASCs accelerated the telogen-to-anagen transition in C3H/HeN mice, and conditioned medium from vitamin C-treated ASCs increased the hair length and the Ki67-positive matrix keratinocytes in hair organ culture. Vitamin C increased the mRNA expression of HGF, IGFBP6, VEGF, bFGF, and KGF, which may mediate hair growth promotion. In summary, vitamin C is transported via SVCT2, and increased ASC proliferation is mediated by the MAPK pathway. In addition, vitamin C preconditioning enhanced the hair growth promoting effect of ASCs. Because vitamin C is safe and effective, it could be used to increase the yield and regenerative potential of ASCs.


Hyperglycemia-impaired aortic vasorelaxation mediated through arginase elevation: Role of stress kinase pathways.
Diabetes-induced vascular endothelial dysfunction has been reported to involve hyperglycemia-induced increases in arginase activity. However, upstream mediators of this effect are not clear. Here, we have tested involvement of Rho kinase, ERK1/2 and p38 MAPK pathways in this process. Studies were performed with aortas isolated from wild type or hemizygous arginase 1 knockout (Arg1+/-) mice and bovine aortic endothelial cells exposed to high glucose (HG, 25 mmol/l) or normal glucose (NG, 5.5 mmol/l) conditions for different times. Effects of inhibitors of arginase, p38 MAPK, ERK1/2 or ROCK and ex vivo adenoviral delivery of active Arg1 and inactive (D128-Arg1) cDNA were also determined. Exposure in wild type aorta or endothelial cells to HG significantly increased arginase activity and Arg1 expression and impaired aortic relaxation. Transduction of wild type aorta with active Arg1 cDNA impaired vascular relaxation, whereas inactive Arg1 had no effect. The HG-induced vascular endothelial dysfunction was associated with increased phosphorylation (activation) of ERK1/2 and p38 MAPK. Pretreatment with inhibitors of ERK1/2, p38 MAPK, ROCK or arginase blocked HG-induced elevation of arginase activity and Arg1 expression and prevented the vascular dysfunction. Inhibition of ROCK blunted the HG-induced activation of ERK1/2 and p38 MAPK. In summary, activated ROCK and subsequent activation of ERK1/2 or p38 MAPK elevates arginase activity and Arg1 expression in hyperglycemic states. Targeting this pathway may provide an effective means for preventing diabetes/hyperglycemia-induced vascular endothelial dysfunction.


Semen quality of consecutive ejaculates from cancer patients for fertility preservation.
To study variations in semen parameters among cancer patients who visited a sperm banking clinic before undergoing cancer treatment. Retrospective, consecutive study. University-based hospital. Eighty-six patients, diagnosed with various cancers, undergoing multiple semen collections on 5 consecutive days, for fertility preservation, between 2004 and 2013. None. Within- and between-subject coefficients of variation were estimated using a random-effects analysis of variance to assess the consistency of semen parameters (volume, sperm concentration, motility, rapid motility, total motile sperm count, and computer-based sperm parameters), whereas intraclass correlation coefficients (ICCs) were calculated to assess the size of the between-component of variance relative to the total component of variance. When analyzing semen parameters over a maximum of 5 consecutive days, only the semen volume was significantly reduced in day-1 and -3 samples compared with the first sample. Almost all of the parameters showed high ICC values, suggesting that within-subject fluctuations were small relative to the between-subject variability. The highest ICC values were noted in volume (ICC 0.76; 95% confidence interval [CI] 0.52-0.89), followed by total motile count (ICC 0.71; 95% CI 0.30-0.89); the least consistent measure was wobble (ICC 0.14; 95% CI -0.13, 0.51). Repeated ejaculates from cancer patients did not show substantial variation in semen quality.


Are fluoride levels in drinking water associated with hypothyroidism prevalence in England? A large observational study of GP practice data and fluoride levels in drinking water.
While previous research has suggested that there is an association between fluoride ingestion and the incidence of hypothyroidism, few population level studies have been undertaken. In England, approximately 10% of the population live in areas with community fluoridation schemes and hypothyroidism prevalence can be assessed from general practice data. This observational study examines the association between levels of fluoride in water supplies with practice level hypothyroidism prevalence. We used a cross-sectional study design using secondary data to develop binary logistic regression models of predictive factors for hypothyroidism prevalence at practice level using 2012 data on fluoride levels in drinking water, 2012/2013 Quality and Outcomes Framework (QOF) diagnosed hypothyroidism prevalence data, 2013 General Practitioner registered patient numbers and 2012 practice level Index of Multiple Deprivation scores. We found that higher levels of fluoride in drinking water provide a useful contribution for predicting prevalence of hypothyroidism. We found that practices located in the West Midlands (a wholly fluoridated area) are nearly twice as likely to report high hypothyroidism prevalence in comparison to Greater Manchester (non-fluoridated area). In many areas of the world, hypothyroidism is a major health concern and in addition to other factors-such as iodine deficiency-fluoride exposure should be considered as a contributing factor. The findings of the study raise particular concerns about the validity of community fluoridation as a safe public health measure.


Indolyl Septanoside Synthesis for In Vivo Screening of Bacterial Septanoside Hydrolases.
Building-up and breaking-down of carbohydrates are processes common to all forms of life. Glycoside hydrolases are a broad class of enzymes that play a central role in the cleavage of glycosidic bonds, which is fundamental to carbohydrate degradation. The large majority of substrates are five- and six-membered ring glycosides. Our interest in seven-membered ring septanose sugars has inspired the development of a way to search for septanoside hydrolase activity. Described here is a strategy for the discovery of septanoside hydrolases that uses synthetic indolyl septanosides as chromogenic substrates. Access to these tool compounds was enabled by a route where septanosyl halides act as glycosyl donors for the synthesis of the indolyl septanosides. The screening strategy leverages the known dimerization of 3-hydroxy-indoles to make colored dyes, as occurs when the β-galactosidase substrate X-Gal is hydrolyzed. Because screens in bacterial cells would enable searches in organisms that utilize heptoses or from metagenomics libraries, we also demonstrate that septanosides are capable of entering E. coli cells through the use of a BODIPY-labeled septanoside. The modularity of the indolyl septanoside synthesis should allow the screening of a variety of substrates that mimic natural structures via this general approach.


Fish in a ring: spatio-temporal pattern formation in one-dimensional animal groups.
In this work, we study the collective behaviour of fish shoals in annular domains. Shoal mates are modelled as self-propelled particles moving on a discrete lattice. Collective decision-making is determined by information exchange among neighbours. Neighbourhoods are specified using the perceptual limit and numerosity of fish. Fish self-propulsion and obedience to group decisions are described through random variables. Spatio-temporal schooling patterns are measured using coarse observables adapted from the literature on coupled oscillator networks and features of the time-varying network describing the fish-to-fish information exchange. Experiments on zebrafish schooling in an annular tank are used to validate the model. Effects of group size and obedience parameter on coarse observables and network features are explored to understand the implications of perceptual numerosity and spatial density on fish schooling. The proposed model is also compared with a more traditional metric model, in which the numerosity constraint is released and fish interactions depend only on physical configurations. Comparison shows that the topological regime on which the proposed model is constructed allows for interpreting characteristic behaviours observed in the experimental study that are not captured by the metric model.


Modification of glial response in hibernation: a patch-clamp study on glial cells acutely isolated from hibernating land snail.
Hibernation is a dormant state of some animal species that enables them to survive harsh environmental conditions during the winter seasons. In the hibernating state, preservation of neuronal rhythmic activity at a low level is necessary for maintenance of suspended forms of behavior. As glial cells support rhythmic activity of neurons, preservation of brain function in the hibernating state implies accompanying modification of glial activity. A supportive role of glia in regulating neuronal activity is reflected through the activity of inwardly rectifying K+ channels (Kir). Therefore, we examined electrophysiological response, particularly Kir current response, of glial cells in mixture with neurons acutely isolated from active and hibernating land snail Helix pomatia. Our data show that hibernated glia have significantly lower inward current density, specific membrane conductance, and conductance density compared with active glia. The observed reduction could be attributed to the Kir currents, since the Ba2+-sensitive Kir current density was significantly lower in hibernated glia. Accordingly, a significant positive shift of the current reversal potential indicated a more depolarized state of hibernated glia. Data obtained show that modification of glial current response could be regulated by serotonin (5-HT) through an increase of cGMP as a secondary messenger, since extracellular addition of 5-HT or intracellular administration of cGMP to active glia induced a significant reduction of inward current density and thus mimicked the reduced response of hibernated glia. Lower Kir current density of hibernated glia accompanied the lower electrical activity of hibernated neurons, as revealed by a decrease in neuronal fast inward Na+ current density. Our findings reveal that glial response is reduced in the hibernating state and suggest seasonal modulation of glial activity. Maintenance of low glial activity in hibernation could be important for preservation of brain rhythmic activity and survival of the animal.


HDAC2 provides a critical support to malignant progression of hepatocellular carcinoma through feedback control of mTORC1 and AKT.
Aberrant regulation of histone deacetylase 2 (HDAC2) contributes to malignant progression in various cancers, but the underlying mechanism leading to the activation of oncogenic HDAC2 remains unknown. In this study, we show that HDAC2 expression is upregulated in a large cohort of patients with human hepatocellular carcinoma, and that high expression of HDAC2 was significantly associated with poor prognosis of patients with hepatocellular carcinoma. We found that mTORC1/NF-κBp50 signaling is necessary for the growth factor-induced HDAC2 and is sustained in hepatocellular carcinoma, but not in normal hepatic cells. Growth factor-induced mTORC1 activates the nuclear translocation of NF-κBp50, where it binds to the intragenic sequences of the HDAC2 gene and promotes its transcription. Hepatocellular carcinoma tissues derived from chemical-induced mouse and rat liver cancer models validated that mTORC1 activation and NF-κBp50 nuclear translocation are essential for the transcriptional activation of oncogenic HDAC2 in hepatocellular carcinoma. In addition, we demonstrate that HDAC2 is required to maintain mTORC1 activity by stabilizing the mTOR/RAPTOR complex. Elevated expression of HDAC2 triggers a positive feedback loop that activates AKT phosphorylation via the transcriptional modulation of phosphoinositide signaling molecules. Bioinformatics analysis of HDAC2 signature and immunoblot analysis of mesenchymal genes also evidenced that HDAC2 plays a role in the malignant behavior of tumor cells by Snail induction and simultaneously E-cadherin suppression in hepatocellular carcinoma cells. These findings establish a molecular mechanism responsible for the activation of oncogenic HDAC2, which explains how growth factor-induced HDAC2 maintains mitogenic signaling and function during hepatocellular malignant progression and provide a novel strategy for therapeutic intervention in liver cancer. Cancer Res; 74(6); 1728-38. ©2014 AACR.


The 'in vivo lifestyle' of bile acid 7α-dehydroxylating bacteria: comparative genomics, metatranscriptomic, and bile acid metabolomics analysis of a defined microbial community in gnotobiotic mice.
The formation of secondary bile acids by gut microbes is a current topic of considerable biomedical interest. However, a detailed understanding of the biology of anaerobic bacteria in the genus Clostridium that are capable of generating secondary bile acids is lacking. We therefore sought to determine the transcriptional responses of two prominent secondary bile acid producing bacteria, Clostridium hylemonae and Clostridium hiranonis to bile salts (in vitro) and the cecal environment of gnotobiotic mice. The genomes of C. hylemonae DSM 15053 and C. hiranonis DSM 13275 were closed, and found to encode 3,647 genes (3,584 protein-coding) and 2,363 predicted genes (of which 2,239 are protein-coding), respectively, and 1,035 orthologs were shared between C. hylemonae and C. hiranonis. RNA-Seq analysis was performed in growth medium alone, and in the presence of cholic acid (CA) and deoxycholic acid (DCA). Growth with CA resulted in differential expression (>0.58 log2FC; FDR < 0.05) of 197 genes in C. hiranonis and 118 genes in C. hylemonae. The bile acid-inducible operons (bai) from each organism were highly upregulated in the presence of CA but not DCA. We then colonized germ-free mice with human gut bacterial isolates capable of metabolizing taurine-conjugated bile acids. This consortium included bile salt hydrolase-expressing Bacteroides uniformis ATCC 8492, Bacteroides vulgatus ATCC 8482, Parabacteroides distasonis DSM 20701, as well as taurine-respiring Bilophila wadsworthia DSM 11045, and deoxycholic/lithocholic acid generating Clostridium hylemonae DSM 15053 and Clostridium hiranonis DSM 13275. Butyrate and iso-bile acid-forming Blautia producta ATCC 27340 was also included. The Bacteroidetes made up 84.71% of 16S rDNA cecal reads, B. wadsworthia, constituted 14.7%, and the clostridia made up <.75% of 16S rDNA cecal reads. Bile acid metabolomics of the cecum, serum, and liver indicate that the synthetic community were capable of functional bile salt deconjugation, oxidation/isomerization, and 7α-dehydroxylation of bile acids. Cecal metatranscriptome analysis revealed expression of genes involved in metabolism of taurine-conjugated bile acids. The in vivo transcriptomes of C. hylemonae and C. hiranonis suggest fermentation of simple sugars and utilization of amino acids glycine and proline as electron acceptors. Genes predicted to be involved in trimethylamine (TMA) formation were also expressed.


Artocarpin induces cell apoptosis in human osteosarcoma cells through endoplasmic reticulum stress and reactive oxygen species.
Osteosarcoma is a malignant primary bone tumor that responds poorly to both chemotherapy and radiation therapy. However, because of side effects and drug resistance in chemotherapy and the insufficiency of an effective adjuvant therapy for osteosarcoma, it is necessary to research novel treatments. This study was the first to investigate the anticancer effects of the flavonoid derivative artocarpin in osteosarcoma. Artocarpin induced cell apoptosis in three human osteosarcoma cell lines-U2OS, MG63, and HOS. Artocarpin was also associated with increased intracellular reactive oxygen species (ROS). Mitochondrial dysfunction was followed by the release of cytochrome c from mitochondria and accompanied by decreased antiapoptotic Bcl-2 and Bcl-xL and increased proapoptotic protein Bak and Bax. Artocarpin triggered endoplasmic reticulum (ER) stress, as indicated by changes in cytosol calcium levels and increased glucose-regulated protein 78 and 94 expressions, and also increased calpains expression and activity. Animal studies revealed a dramatic 40% reduction in tumor volume after 18 days of treatment. This study demonstrated a novel anticancer activity of artocarpin against human osteosarcoma cells and in murine tumor models. In summary, artocarpin significantly induced cell apoptosis through ROS, ER stress, mitochondria, and the caspase pathway, and may thus be a novel anticancer treatment for osteosarcoma.


The impact of tax policies on living organ donations in the United States.
In an effort to increase living organ donation, fifteen states passed tax deductions and one a tax credit to help defray potential medical, lodging and wage loss costs between 2004 and 2008. To assess the impact of these policies on living donation rates, we used a differences-in-differences strategy that compares the pre- and postlegislation change in living donations in states that passed legislation against the same change in those states that did not. We found no statistically significant effect of these tax policies on donation rates. Furthermore, we found no evidence of any lagged effects, differential impacts by gender, race or donor relationship, or impacts on deceased donation. Possible hypotheses to explain our findings are: the cash value of the tax deduction may be too low to defray costs faced by donors, lack of public awareness about the existence of these policies, and that states that were proactive enough to pass tax policy laws may have already depleted donor pools with previous interventions.


Job crafting in changing organizations: Antecedents and implications for exhaustion and performance.
The present study addressed employee job crafting behaviors (i.e., seeking resources, seeking challenges, and reducing demands) in the context of organizational change. We examined predictors of job crafting both at the organizational level (i.e., perceived impact of the implemented changes on the working life of employees) and the individual level (i.e., employee willingness to follow the changes). Job crafting behaviors were expected to predict task performance and exhaustion. Two-wave longitudinal data from 580 police officers undergoing organizational changes were analyzed with structural equation modeling. Findings showed that the degree to which changes influence employees' daily work was linked to reducing demands and exhaustion, whereas employee willingness to change was linked to seeking resources and seeking challenges. Furthermore, while seeking resources and seeking challenges were associated with high task performance and low exhaustion respectively, reducing demands seemed to predict exhaustion positively. Our findings suggest that job crafting can act as a strategy of employees to respond to organizational change. While seeking resources and seeking challenges enhance employee adjustment and should be encouraged by managers, reducing demands seems to have unfavorable implications for employees.


Uterine torsion in pregnancy. Case Report.
This paper describes a case of uterine torsion not only around the long, but also the horizontal axis, diagnosed post-operatively. The patient was 28-years-old, II/I in the 34th week of her gravidity. She was admitted due to amniotic fluid leak. Due to unsuccessful delivery induction using prostaglandins, a caesarean section was indicated to end the gravidity. Uterine torsion 180° around the longitudinal axis and 90° around the transversal axis was diagnosed during the surgery. The uterine wound was treated in accordance with guidelines and the patient was dismissed in a good condition. Control hysteroscopy and laparoscopy done after a time interval showed problem-free healing of the wound in the posterior uterine wall.


Velocimetry microsensors driven by linearly polarized optical tweezers.
We report on a new type of single-point velocimetry microsensor that can be positioned in microfluidic devices by use of optical tweezers. The flag-shaped microsensor is readily made by a low-cost two-photon polymerization technique. At rest the linearly polarized optical tweezer traps the microsensor at the focal point, and the flag-plate gets aligned in the polarization direction. Under a fluid flow, the plate rotates to an equilibrium angle that is used to measure the fluid velocity with a micrometer-size spatial resolution. Experimental results are in good agreement with theoretical calculations of optical and hydrodynamic torques on such a flag-shaped microsensor.


Cultivation of an obligate Fe(II)-oxidizing lithoautotrophic bacterium using electrodes.
Fe(II)-oxidizing aerobic bacteria are poorly understood, due in part to the difficulties involved in laboratory cultivation. Specific challenges include (i) providing a steady supply of electrons as Fe(II) while (ii) managing rapid formation of insoluble Fe(III) oxide precipitates and (iii) maintaining oxygen concentrations in the micromolar range to minimize abiotic Fe(II) oxidation. Electrochemical approaches offer an opportunity to study bacteria that require problematic electron donors or acceptors in their respiration. In the case of Fe(II)-oxidizing bacteria, if the electron transport machinery is able to oxidize metals at the outer cell surface, electrodes poised at potentials near those of natural substrates could serve as electron donors, eliminating concentration issues, side reactions, and mineral end products associated with metal oxidation. To test this hypothesis, the marine isolate Mariprofundus ferrooxydans PV-1, a neutrophilic obligate Fe(II)-oxidizing autotroph, was cultured using a poised electrode as the sole energy source. When cells grown in Fe(II)-containing medium were transferred into a three-electrode electrochemical cell, a cathodic (negative) current representing electron uptake by bacteria was detected, and it increased over a period of weeks. Cultures scraped from a portion of the electrode and transferred into sterile reactors consumed electrons at a similar rate. After three transfers in the absence of Fe(II), electrode-grown biofilms were studied to determine the relationship between donor redox potential and respiration rate. Electron microscopy revealed that under these conditions, M. ferrooxydans PV-1 attaches to electrodes and does not produce characteristic iron oxide stalks but still appears to exhibit bifurcate cell division. IMPORTANCE Electrochemical cultivation, supporting growth of bacteria with a constant supply of electron donors or acceptors, is a promising tool for studying lithotrophic species in the laboratory. Major pitfalls present in standard cultivation methods used for metal-oxidizing microbes can be avoided by the use of an electrode as the sole electron donor. Electrochemical cultivation also offers a window into the poorly understood metabolism of microbes such as obligate Fe(II), Mn(II), or S(0) oxidizers by replacing the electron source with the controlled surface of an electrode. The elucidation of redox-dependent behavior of these microbes could enhance industrial applications tuned to oxidation of specific metals, provide insight into how bacteria evolved to compete with oxygen for reactive metal species, and model geochemical impacts of their metabolism in the environment.


Drugs for preventing red blood cell dehydration in people with sickle cell disease.
Sickle cell disease is an inherited disorder of haemoglobin, which results in abnormal red blood cells. These can deform and cause blockages in blood vessels, leading to acute crises such as pain, stroke and splenic sequestration, and chronic organ and tissue damage. Recently research has begun to focus on therapies which prevent the red blood cells deforming by reducing the loss of water and ions from the cells. However, little is known about the effectiveness and safety of such drugs. To assess the relative risks and benefits of drugs which aim to prevent sickle cell related crises by reducing red blood cell dehydration. We searched the Cochrane Cystic Fibrosis and Genetic Disorders Group specialist register which comprises references identified from comprehensive electronic database searches, handsearching relevant journals and handsearching abstract books of conference proceedings. Date of the most recent search of the Group's specialised register: December 2001. All those randomised or quasi-randomised controlled trials of drugs which aim to prevent sickle cell crises by reducing red cell dehydration, compared to placebo or an alternative treatment. Both reviewers independently selected trials for inclusion, assessed trial quality and extracted data from the included studies. Of the 27 trials identified, two met the inclusion criteria. The two trials tested the effectiveness of zinc sulphate and piracetam to prevent sickle cell related crises in a total of 246 patients. A reduction in pain crises was shown in the piracetam study over one year (weighted mean difference (WMD) -1.9 (95% CI -3.01, -0.79)), although blood counts were not significantly changed. The zinc trial showed a significant reduction in the total number of pain, haemolytic, aplastic and sequestration crises over one and a half years (WMD -2.83 (95% CI -3.51, -2.15)), but our analysis was limited by non-reporting of standard deviations for some data. Changes to red cell parameters and blood counts were inconsistent. No serious adverse events were noted in either trial. While the results of both zinc and piracetam for reducing sickle related crises are encouraging, larger, and/or longer term multicentre trials over a number of years are needed to evaluate the effectiveness of these therapies for patients with sickle cell disease.


Use of prior vaccinations for the development of new vaccines.
There is currently a need for vaccine development to improve the immunogenicity of protective epitopes, which themselves are often poorly immunogenic. Although the immunogenicity of these epitopes can be enhanced by linking them to highly immunogenic carriers, such carriers derived from current vaccines have not proven to be generally effective. One reason may be related to epitope-specific suppression, in which prior vaccination with a protein can inhibit the antibody response to new epitopes linked to the protein. To circumvent such inhibition, a peptide from tetanus toxoid was identified that, when linked to a B cell epitope and injected into tetanus toxoid-primed recipients, retained sequences for carrier but not suppressor function. The antibody response to the B cell epitope was enhanced. This may be a general method for taking advantage of previous vaccinations in the development of new vaccines.


Tumor necrosis factor system activity is associated with insulin resistance and dyslipidemia in myotonic dystrophy.
Myotonic dystrophy (MyD) is a multisystem autosomal dominant disorder associated with progressive muscle wasting and weakness. The striking metabolic abnormality in MyD is insulin resistance. The mechanism by which target tissues are insensitive to insulin action remains uncertain. In a recent study, plasma soluble tumor necrosis factor receptor (sTNFR)2 levels were found to be associated with muscle tissue mass and insulin resistance. Given these associations, we speculated that disorders of the muscle cell membrane could lead simultaneously to insulin insensitivity and sTNFR2 leakage in MyD. To test this hypothesis, we measured the levels of circulating sTNFR1 and sTNFR2 and insulin resistance in MyD patients. We studied 22 MyD patients and 24 age-, BMI-, and fat mass-matched control subjects. Both MyD men and women showed higher plasma insulin levels in the presence of comparable glucose concentrations than did control subjects. sTNFR2, but not sTNFR1, levels were approximately 1.5-fold higher in MyD patients. In parallel with these findings, the fasting insulin resistance index (FIRI) was also higher in MyD patients. In fact, in the whole population, fasting insulin and FIRI strongly correlated with sTNFR2 in both men (r = 0.77 and r = 0.81, P<0.0001, respectively) and women (r = 0.67 and r = 0.64, P = 0.001, respectively). sTNFR2 levels were also associated with the insulin sensitivity index (S(I)), calculated from an oral glucose tolerance test (OGTT) according to the method by Cederholm and Wibell (r = -0.43, P = 0.006). We constructed a multiple linear regression to predict FIRI, with BMI, waist-to-hip ratio, and sTNFR2 as independent variables. In this model, both BMI (P = 0.0014) and sTNFR2 (P = 0.0048) levels contributed independently to 46% of the variance of FIRI. In another model, in which FIRI was substituted for S(I) from the OGTT, both BMI (P = 0.0001) and sTNFR2 (P = 0.04) levels contributed independently to 48% of the variance of S(I) from the OGTT. Plasma cholesterol and triglyceride concentrations were significantly increased in MyD patients. sTNFR1 and sTNFR2 levels were found to be strongly associated with plasma cholesterol, LDL cholesterol, and triglycerides. sTNFR1 and sTNFR2 also correlated with serum creatine kinase activity in MyD patients (r = 0.57, P = 0.006; r = 0.75, P<0.0001, respectively). In conclusion, here we describe, for the first time to our knowledge, a relationship between insulin action and plasma sTNFR2 concentration in MyD patients. We have also found increased concentrations of plasma triglycerides and cholesterol levels in parallel with sTNFR1 and sTNFR2 concentrations in MyD patients. We speculate that the latter associations are dependent on, and secondary to, increased tumor necrosis factor (TNF)-alpha action. Whether TNF action is implicated in the pathogenesis of MyD or is a simple marker of disease activity awaits further studies.


Prospective memory in clinical populations.
Prospective memory (PM) has emerged as a form of episodic memory that is frequently impaired in a variety of clinical populations. Neuropsychologists who routinely evaluate these populations are often unaware of the possibility of PM deficits or the impact these deficits may have on everyday functioning. The objective of this special issue is to provide an overview of the nature of prospective deficits in a range of clinical populations, to discuss neuropsychological assessment techniques, and to critically evaluate management strategies. We solicited papers from established researchers and issued a general call for papers for the special issue on PM in clinical populations. We received submissions from the nine authors that we solicited. These submissions range from developmental disorders, including autism, attention deficit hyperactivity disorder, and dyslexia; to disorders of adulthood, such as schizophrenia, HIV, brain injury, and multiple sclerosis; and finally disorders that tend to occur at older ages, such as Parkinson's disease and mild cognitive impairment. In addition, we have included four original research articles that provide novel data on other populations. These are children and adolescents with 22q11.2 deletion syndrome, first-degree relatives of people with schizophrenia, individuals with mild brain injury, and individuals with idiopathic REM sleep behavioral disorder. The issue highlights the need for clinical neuropsychologists to be aware of the possible existence of deficits in PM in a variety of clinical populations and the importance of both assessment and management strategies to reduce the impact on daily life.


Race/Ethnicity and Sexual Orientation Disparities in Mental Health, Sexism, and Social Support among Women Veterans.
To identify patterns of risk and resilience by the intersections of race/ethnicity and sexual orientation in mental health symptom severity, sexism, and social support among U.S. women veterans. A national sample of women veterans (n = 648, 38% sexual minority, 15% racial/ethnic minority) was recruited online in 2013 using social networking websites and listservs. Using cross-sectional survey data, we evaluated main and interactive associations between race/ethnicity and sexual orientation on depression, anxiety, posttraumatic stress, unhealthy alcohol use, sexism, and social support. Models were adjusted for other demographic characteristics. Across depression, anxiety, and sexism, White heterosexual women reported the least distress and racial/ethnic minority heterosexual women the most distress (race/ethnicity x sexual orientation interactions p < .05). Among White women, sexual minority women reported greater levels of depression, anxiety, and sexism than heterosexual women. The effects were the opposite among racial/ethnic minority women, where heterosexual women reported similar or worse depression, anxiety, and sexism than sexual minority women. There were no race/ethnicity or sexual orientation interaction effects on posttraumatic stress symptoms or unhealthy alcohol use and marginally significant effects on social support. Among women veterans, race/ethnicity and sexual orientation were associated with mental health and sexism, alone and in combination. Findings suggest that those who were both racial/ethnic and sexual minorities may develop resilience from their lived experience. On the other hand, women veterans with a minority race/ethnicity or a minority sexual orientation appeared more vulnerable to adverse outcomes and may need targeted care.


T helper type 1 and 17 cells determine efficacy of interferon-beta in multiple sclerosis and experimental encephalomyelitis.
Interferon-beta (IFN-beta) is the major treatment for multiple sclerosis. However, this treatment is not always effective. Here we have found congruence in outcome between responses to IFN-beta in experimental autoimmune encephalomyelitis (EAE) and relapsing-remitting multiple sclerosis (RRMS). IFN-beta was effective in reducing EAE symptoms induced by T helper type 1 (T(H)1) cells but exacerbated disease induced by T(H)17 cells. Effective treatment in T(H)1-induced EAE correlated with increased interleukin-10 (IL-10) production by splenocytes. In T(H)17-induced disease, the amount of IL-10 was unaltered by treatment, although, unexpectedly, IFN-beta treatment still reduced IL-17 production without benefit. Both inhibition of IL-17 and induction of IL-10 depended on IFN-gamma. In the absence of IFN-gamma signaling, IFN-beta therapy was ineffective in EAE. In RRMS patients, IFN-beta nonresponders had higher IL-17F concentrations in serum compared to responders. Nonresponders had worse disease with more steroid usage and more relapses than did responders. Hence, IFN-beta is proinflammatory in T(H)17-induced EAE. Moreover, a high IL-17F concentration in the serum of people with RRMS is associated with nonresponsiveness to therapy with IFN-beta.


Spinal subarachnoid cysts in 13 dogs.
Thirteen dogs, including 6 Rottweiler dogs, exhibiting clinical signs of spinal cord dysfunction and myelographically confirmed subarachnoid space enlargement were investigated. To characterize the lesions and to get a better understanding of their pathogenesis, different imaging techniques were used in association with explorative surgical procedures (12 dogs) and histopathologic techniques (5 dogs). All subjects underwent preoperative myelography, five of which were examined by computed tomography (CT) scanning and one by magnetic resonance imaging (MRI) as well as cerebrospinal fluid (CSF) flow measurement (velocimetry). Most animals were <12 months old (7/13 dogs) and Rottweilers were over-represented (6/13 dogs). The lesions were mainly located dorsally with respect to the spinal cord (10/13 dogs) and in the cranial cervical area (8/13 dogs). MRI suggested spinal cord deviation with signs of ventral leptomeningeal adhesion opposite the enlarged space. In one dog, velocimetry confirmed that the "cyst" was freely communicating with the surrounding CSF space. Surgical investigation confirmed leptomeninges-induced ventral adhesion in 4/5 dogs. Follow-up studies, carried out from 6 months to 2.5 years postoperatively, showed there was full recovery in 8/13 dogs. This study suggests that the compression of the spinal cord is possibly not caused by a cyst. Adhesion resulting from a combination of microtrauma and chronic inflammatory processes induces a secondary enlargement of the subarachnoid space and may be a significant causative factor in spinal cord compression and dysfunction. The over-representation of Rottweilers and the young age of the animals in the study suggest a possible genetic predisposition and an inherited etiology.


A phase I randomized study of subcutaneous adjuvant IL-2 in combination with an autologous tumor vaccine in patients with advanced renal cell carcinoma.
We performed a prospective, randomized study to determine whether subcutaneous administration of interleukin-2 (IL-2) in combination with an autologous renal cell vaccine is feasible and can potentiate antitumor immunity. Seventeen patients with metastatic renal cell carcinoma underwent surgical resection with preparation of an autologous tumor cell vaccine. Patients were vaccinated intradermally twice at weakly intervals with 10(7) irradiated tumor cells plus bacillus Calmette-Guérin, and once with 10(7) tumor cells alone. Patients were randomized to one of three groups: no adjuvant IL-2, low-dose IL-2 (1.2 x 10(6) IU/m2), or high-dose IL-2 (1.2 x 10(7) IU/m2). IL-2 was administered subcutaneously on the day of vaccination and the subsequent 4 days. Immune response was monitored by delayed-type hypersensitivity (DTH) response to tumor cells as compared with normal autologous renal cells. Sixteen of 17 patients received vaccine therapy. Four patients developed cellular immunity specific for autologous tumor cells as measured by DTH responses; two had received no IL-2 and two had received high-dose IL-2. There were two partial responses (PR) noted, both in patients who received high-dose IL-2. One responding patient was DTH(+) and one was negative. A third patient who was DTH(+) after vaccination with no IL-2 had a dramatic PR after receiving IL-2 subcutaneously in a subsequent protocol. Prospective testing of response to recall antigens indicated that only 5 of 12 tested patients were positive, including both clinical responders. These data suggest that subcutaneously administered adjuvant IL-2 does not dramatically augment the immunologic response to autologous renal cell vaccines as determined by the development of tumor-specific DTH response.


Traceable calibration, performance metrics, and uncertainty estimates of minirhizotron digital imagery for fine-root measurements.
Even though fine-root turnover is a highly studied topic, it is often poorly understood as a result of uncertainties inherent in its sampling, e.g., quantifying spatial and temporal variability. While many methods exist to quantify fine-root turnover, use of minirhizotrons has increased over the last two decades, making sensor errors another source of uncertainty. Currently, no standardized methodology exists to test and compare minirhizotron camera capability, imagery, and performance. This paper presents a reproducible, laboratory-based method by which minirhizotron cameras can be tested and validated in a traceable manner. The performance of camera characteristics was identified and test criteria were developed: we quantified the precision of camera location for successive images, estimated the trueness and precision of each camera's ability to quantify root diameter and root color, and also assessed the influence of heat dissipation introduced by the minirhizotron cameras and electrical components. We report detailed and defensible metrology analyses that examine the performance of two commercially available minirhizotron cameras. These cameras performed differently with regard to the various test criteria and uncertainty analyses. We recommend a defensible metrology approach to quantify the performance of minirhizotron camera characteristics and determine sensor-related measurement uncertainties prior to field use. This approach is also extensible to other digital imagery technologies. In turn, these approaches facilitate a greater understanding of measurement uncertainties (signal-to-noise ratio) inherent in the camera performance and allow such uncertainties to be quantified and mitigated so that estimates of fine-root turnover can be more confidently quantified.


Ketones and Human Performance.
Everyone is seeking nutritional strategies that might benefit performance. One approach receiving much attention is ketones, or ketosis. Ketones are very simple compounds made of hydrogen, carbon, and oxygen, and ketosis is a metabolic state whereby the body uses predominantly ketones. Ketosis can be achieved by fasting for longer than 72 hours or by following a very lowcarbohydrate, high-fat diet (ketogenic diet) for several days to weeks. Alternatively, ketone supplements purportedly induce ketosis rapidly and do not require strict adherence to any specific type of diet; however, much of the touted benefits are anecdotal. A potential role for ketosis as a performance enhancer was first introduced in 1983 with the idea that chronic ketosis without caloric restriction could preserve submaximal exercise capability by sparing glycogen or conserving the limited carbohydrate stores. Few human studies on the effects of a ketogenic diet on performance have yielded positive results, and most studies have yielded equivocal or null results, and a few negative results. Many questions about ketones relevant to Special Operations Forces (SOF) remain unanswered. At present, a ketogenic diet and/or a ketone supplement do not appear confer performance benefits for SOF. Instead, Operators should engage with their unit dietitian to develop individualized nutritional strategies based on unique mission requirements. The authors review the concept of a ketogenic diet, describe some potential benefits and risks of ketosis, review the performance literature and how to measure ketone status, and then summarize the landscape in 2017.


Suicide after bereavement: an overlooked problem.
To examine the effect of time on suicide after bereavement among widowed persons. The data were extracted from Swiss mortality statistics for the period 1987-2005. The time between bereavement and subsequent death, specifically by suicide, was determined by linkage of individual records of married persons. The suicide rates and the standardized mortality ratios in the first week/month/year of widowhood were calculated based on person-year calculations. The annualized suicide rates in widowed persons were highest in the first week after bereavement: 941 males and 207 females per 100,000. The corresponding standardized mortality ratios were approximately 34 and 19 respectively. In the first month(s) after bereavement, the rates and the ratios decreased, first rapidly, then gradually. Except in older widows, they did not reach the baseline levels during the first year after bereavement. The suicide risk of widowed persons is increased in the days, weeks and months after bereavement. Widowed persons are a clear-cut risk group under the aegis of undertakers, priests and general practitioners.


Apoptotic neutrophils containing Staphylococcus epidermidis stimulate macrophages to release the proinflammatory cytokines tumor necrosis factor-alpha and interleukin-6.
Staphylococcus epidermidis infections are usually nosocomial and involve colonization of biomaterials. The immune defense system cannot efficiently control the bacteria during these infections, which often results in protracted chronic inflammation, in which a key event is disturbed removal of neutrophils by tissue macrophages. While ingesting uninfected apoptotic neutrophils, macrophages release anti-inflammatory cytokines that lead to resolution of inflammation. In clinical studies, we have previously found elevated levels of the proinflammatory cytokines tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 in synovial fluid from prostheses infected with coagulase negative staphylococci. We show that macrophages phagocytosing apoptotic neutrophils containing S. epidermidis released TNF-alpha and interleukin-6, whereas macrophages phagocytosing spontaneously apoptotic neutrophils did not. This difference was not due to dissimilar phagocytic capacities, because macrophages ingested both types of neutrophils to the same extent. The activation was induced mainly by the apoptotic neutrophils themselves, not by the few remaining extracellular bacteria. Macrophages were not activated by apoptotic neutrophils that contained paraformaldehyde-killed S. epidermidis. Proinflammatory reactions induced by clearance of apoptotic neutrophils containing S. epidermidis might represent an important mechanism to combat the infective agent. This activation of macrophages may contribute to the development of chronic inflammation instead of inflammation resolution.


A UVC device for intra-luminal disinfection of catheters: in vitro tests on soft polymer tubes contaminated with Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and Candida albicans.
Bacterial colonization of central venous catheters (CVCs) causes severe complications in patients. As a result, developing methods to remove and prevent bacterial and fungal colonization of CVCs is imperative. Recently, we have demonstrated that disinfection by radiation of polymer tubes with UVC light is possible. In this paper we present dose-response results using a newly developed UVC disinfection device, which can be connected to a Luer catheter hub. The device was tested on soft polymer tubes contaminated with a pallet of microorganisms, including Candida albicans, Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa (ca 10(3) CFU mL(-1)). The tubes were equipped with a modified catheter hub and interfaced to the disinfection device via a middle piece separating the disinfection device from the hub. The contamination lasted for 3 h prior to treatment to simulate an aseptic breach. Our results show UVC killing in a dose and time dependent manner, with no viable counts after 2 min of radiation for bacteria. Killing of C. albicans was obtained at >20 min in an UVC absorbing suspension. We believe our results to be transferable directly to the clinic, and we are currently working on a setup for clinical trial.


EUS anatomy of the liver segments.
Clinical applications of EUS for the liver have been recently increasing. They include the screening and diagnosis of liver parenchymal disease and malignant tumors as well as EUS-guided interventions such as hepaticogastrostomy, tumor ablation therapy, and portal pressure gradient measurement. Although the segmental localization of the targeted tumor, bile duct, and vessel in the liver is important to complete these procedures, little information is available regarding hepatic segmental anatomy on EUS. The liver can be visualized with EUS by transgastric and transduodenal scanning, and the EUS determination of segmental location can be achieved using various anatomical landmarks. Identification of the right posterior segments is, however, technically difficult because they are located far from the stomach and duodenum. In the present review, we describe the normal anatomy of liver segments using linear EUS.


The X-Ray Emission Effectiveness of Plasma Mirrors: Reexamining Power-Law Scaling for Relativistic High-Order Harmonic Generation.
Ultrashort pulsed lasers provide uniquely detailed access to the ultrafast dynamics of physical, chemical, and biological systems, but only a handful of wavelengths are directly produced by solid-state lasers, necessitating efficient high-power frequency conversion. Relativistic plasma mirrors generate broadband power-law spectra, that may span the gap between petawatt-class infrared laser facilities and x-ray free-electron lasers; despite substantial theoretical work the ultimate efficiency of this relativistic high-order-harmonic generation remains unclear. We show that the coherent radiation emitted by plasma mirrors follows a power-law distribution of energy over frequency with an exponent that, even in the ultrarelativistic limit, strongly depends on the ratio of laser intensity to plasma density and exceeds the frequently quoted value of -8/3 over a wide range of parameters. The coherent synchrotron emission model, when adequately corrected for the finite width of emitting electron bunches, is not just valid for p-polarized light and thin foil targets, but generally describes relativistic harmonic generation, including at normal incidence and with finite-gradient plasmas. Our numerical results support the ω-4/3 scaling of the synchrotron emission model as a limiting efficiency of the process under most conditions. The highest frequencies that can be generated with this scaling are usually restricted by the width of the emitting electron bunch rather than the Lorentz factor of the fastest electrons. The theoretical scaling relations developed here suggest, for example, that with a 20-PW 800-nm driving laser, 1 TW/harmonic can be produced for 1-keV photons.


Inhibition of premature labor: a multicenter comparison of ritodrine and ethanol.
A randomized controlled study was carried out at three medical centers to compare the efficacy and side effects of ethanol and ritodrine in the treatment of threatened premature labor. One hundred and thirty-five patients judged to be between the twentieth and thirty-sixth week of gestation and presenting with clinical symptoms of premature labor were included. Sixty-seven patients were treated with intravenous infusion of 10 per cent ethanol. Sixty-eight patients were treated with intravenous infusion of ritodrine for 12 hours followed by oral ritodrine. If labor recurred prematurely, up to two additional courses of ethanol or ritodrine were given. Delivery was postponed for more than 72 hours in 49 of 67 patients (73 per cent) with ethanol and in 61 of 68 patients (90 per cent) with ritodrine; this difference was significant. Patients in the ethanol group gained a mean of 27.6 days while patients in the ritodrine group gained a mean of 44.0 days. Fifty-four per cent of the ethanol group and 72 per cent of the ritodrine group carried their infants to 36 weeks of gestation. Five infants in the ethanol group and one infant in the ritodrine group died from respiratory distress syndrome. The most frequent side effect of ethanol were nausea and vomiting. The most frequent side effects of ritodrine were tachycardia and blood pressure changes which were easily controlled by lowering the infusion rate. Ethanol and ritodrine were both found to be effective inhibitors of premature labor with ritodrine giving the most favorable results.


Comment on "Deep-sea temperature and ice volume changes across the Pliocene-Pleistocene climate transitions".
Sosdian and Rosenthal (Reports, 17 July 2009, p. 306) used magnesium/calcium ratios in benthic foraminifera from the North Atlantic to reconstruct past bottom-water temperatures. They suggested that both ice volume change and ice-sheet dynamics played important roles during the late Pliocene and mid-Pleistocene climate transitions. We present evidence that their record of deep ocean temperature is not reliable, thus raising doubts about their conclusions.


Early Detection of Human Epileptic Seizures Based on Intracortical Microelectrode Array Signals.
We examine, for the first time, the use of intracortical microelectrode array (MEA) signals for early detection of human epileptic seizures. 4×4 mm2 96-channel-MEA recordings were obtained during neuro-monitoring preceding resective surgery in five participants. The participant-specific seizure-detection framework consisted of: first, feature extraction from local field potentials (LFPs) and multiunit activity (MUA); second, nonlinear cost-sensitive support vector machine (SVM) classification of ictal and interictal states based on LFP, MUA, and combined LFP-MUA (a SVM was trained for each participant separately); and third, Kalman filter postprocessing of SVM scoring functions. Performance was assessed on data including 17 seizures and 39.0 h interictal and preictal recordings. The use of combined LFP-MUA features resulted in 100% sensitivity with short detection latency (average: 2.7 s; median: 2.5 s) and five false alarms (0.13/h). The average detection performance based on the area under the receiver operating characteristic corresponded to 0.97. Importantly, technically false alarms were related to epileptiform activity, subclinical seizures, and recording artifacts. Extreme gradient boosting classifiers ranked features based on LFP spectral coherence or MUA count among the top features for seizures characterized by spike-wave complexes, whereas features related to LFP power spectra were ranked higher for seizures characterized by sustained gamma LFP oscillations. The combination of intracortical LFP and MUA signals may allow reliable detection of human epileptic seizures by improving latency and false alarm rate. Intracortical MEAs provide promising signals for closed-loop seizure-control systems based on seizure early-detection in people with pharmacologically resistant epilepsies.


Who needs bereavement support? A population based survey of bereavement risk and support need.
This study identifies and describes the profiles of bereavement risk and support needs of a community sample in Australia and tests the fit of the data with the three-tiered public health model for bereavement support. Family members who were bereaved 6-24 months prior to the survey and who were clients of four funeral providers participated (May-July 2013). A postal survey was used to collect information about bereaved people's experience of caring and perceived satisfaction with any bereavement support provided. The questionnaire included a validated risk assessment screening measure for Prolonged Grief Disorder (PG-13). A total of 678 bereaved people responded. The model predicted that 60% of the sample would be low risk, 30% moderate risk, and 10% high risk. Actual figures were very close at 58.4%, 35.2% and 6.4% respectively. The analysis of the demographic characteristics, experience and impact of caring and bereavement, and satisfaction with support received from a variety of sources revealed differential experiences and needs that align with the expectation of low, moderate, and high bereavement support need, as articulated in the public health model. This is the first empirical test of the public health model of bereavement support. As there is a lack of clear evidence to guide development and allocation of bereavement support programs, the findings have the potential to inform the ability of services, community organizations and informal networks to prioritize care according to each level of bereavement need. This is essential to achieve cost-effective and equitable resource allocation.


Universal use of inactivated polio vaccine.
To present an update on the status of poliomyelitis worldwide, number of cases per year, regions most affected by the disease, vaccines currently available, their risks and benefits, monovalent vaccine use, risks of disseminating a mutant virus in the community, progress that has been made in terms of worldwide eradication and the World Health Organization's (WHO) proposals in this transition period between global eradication and the post-eradication period. Data for the period from 1955 to 2005 were searched in MEDLINE, LILACS, The Web, Doctor's Guide, WHO website and the Pan American Health Organization (PAHO) website and text book. In 1988, the WHO established the goal of eradicating the disease and interrupting transmission of the wild virus globally. Since then, there has been a dramatic decline of the disease, although in 2005 there were still some countries considered endemic and others where polio returned on account of imported viruses. The vaccines used worldwide are the classical tOPV and IPV, and in this eradication process, the use of mOPV vaccines has been encouraged in places where only one type of poliovirus circulates. In addition to spreading the virus in the community, the OPV vaccines may, however, cause paralyses by reversal of the neurovirulence process. For a world free of poliomyelitis disease, it would be necessary to interrupt circulation of the virus, which will only be possible if the OPV virus were to be discontinued, in accordance with the WHO proposals for this transition period and the post-eradication period.


Effect of probiotic Lactobacillus rhamnosus GG intervention on global serum lipidomic profiles in healthy adults.
To investigate the effect of three weeks' intervention with a probiotic Lactobacillus rhamnosus GG (LGG) bacteria on global serum lipidomic profiles and evaluate whether the changes in inflammatory variables (CRP, TNF-alpha and IL-6) are reflected in the global lipidomic profiles of healthy adults. We performed UPLC/MS-based global lipidomic platform analysis of serum samples (n = 26) in a substudy of a randomised, double-blind, placebo-controlled 3-wk clinical intervention trial investigating the immunomodulatory effects of probiotics in healthy adults. A total of 407 lipids were identified, corresponding to 13 different lipid classes. Serum samples showed decreases in the levels of lysophosphatidylcholines (LysoGPCho), sphingomyelins (SM) and several glycerophosphatidylcholines (GPCho), while triacylglycerols (TAG) were mainly increased in the probiotic LGG group during the intervention. Among the inflammatory variables, IL-6 was moderately associated by changes in global lipidomic profiles, with the top-ranked lipid associated with IL-6 being the proinflammatory LysoGPCho (20:4). There was a weak association between the lipidomic profiles and the two other inflammatory markers, TNF-alpha and CRP. This was the first study to investigate the effects of probiotic intervention on global lipidomic profiles in humans. There are indications that probiotic LGG intervention may lead to changes in serum global lipid profiles, as reflected in decreased GPCho, LysoGPCho and SM as well as mainly increased TAG.


Reactions of the nitroso and hydroxylamine metabolites of sulfamethoxazole with reduced glutathione. Implications for idiosyncratic toxicity.
N4-oxidation of sulfonamides has been implicated in the pathogenesis of idiosyncratic reactions to these antimicrobials. In vitro toxicity assays employing mononuclear leukocytes as target cells have shown that the toxicity of sulfamethoxazole hydroxylamine (SMX-HA) is inhibited by exogenous glutathione, suggesting that conjugation with glutathione is an important detoxification pathway. However, in these experiments, significant depletion of cellular glutathione only occurred at concentrations of SMX-HA greater than or equal to 300 microM. At concentrations of SMX-HA which produce 50% toxicity in mononuclear leukocytes (approximately 100 microM), there was not a significant loss of glutathione. SMX-HA also caused a small but significant increase in oxidized glutathione concentrations. In cell-free experiments, reduced glutathione (GSH) prevented the autooxidation of SMX-HA to nitrososulfamethoxazole (nitroso-SMX). During this process, oxidized glutathione was formed. GSH rapidly reacted with nitroso-SMX to form a labile semimercaptal conjugate. Physiologically relevant concentrations of GSH (i.e. 1 mM) favored thiolytic cleavage of the semimercaptal to form SMX-HA. Isomerization of the semimercaptal to the more stable sulfinamide occurred at low GSH concentrations. Purified glutathione transferases had no effect on the reaction of SMX-HA with GSH. Therefore, glutathione is important in protecting cells from the toxicity of SMX-HA largely by preventing its further oxidation to nitroso-SMX. Stable glutathione conjugates are likely to be formed only in small quantities under physiological conditions. Conjugation with glutathione would not be expected to be a major pathway for clearance of the hydroxylamine and nitroso metabolites of sulfonamides.


The effect of different interdental cleaning devices on gingival bleeding.
To compare the effectiveness of an oral irrigator (OI) with a prototype jet tip or a standard jet tip to floss as adjunct to daily toothbrushing on gingival bleeding. In this single masked, 3-group parallel, 4-week home use experiment, 108 subjects were randomly assigned to one of three groups: 1) 01 with a prototype jet tip; 2) OI with a standard jet tip; 3) waxed dental floss. All groups used their assigned product once a day as adjunct to twice daily toothbrushing for two minutes with a standard ADA reference toothbrush. Professional instructions were given by a dental hygienist in 01 use or floss use according to written instructions. All subjects also received a toothbrush instruction leaflet (Bass technique). Subjects were assessed for both bleeding and plaque at baseline and after two weeks and four weeks and were instructed to brush their teeth approximately 2 to 3 hours prior to their assessment. With respect to mean bleeding scores the ANCOVA analysis with baseline as covariate and week 4 as dependent variable showed a significant difference between groups in favor of both the oral irrigator groups. For plaque, however, no significant difference among groups was observed. When combined with manual toothbrushing the daily use of an oral irrigator, either with prototype or standard jet tip, is significantly more effective in reducing gingival bleeding scores than is the use of dental floss, as determined within the limits of this 4-week study design.


A location-specific spreadsheet for estimating Zika risk and timing for Zika vector surveillance, using US military facilities as an example.
Local Zika virus transmission in the United States involving one or both of the known vector species, Aedes aegypti and Ae. albopictus, is of major concern. To assist efforts to anticipate the risks of transmission, we developed an Excel spreadsheet tool that uses vector and virus temperature thresholds, remotely sensed maximum temperature, and habitat suitability models to answer the questions: "is Zika transmission likely here?" and "when should we conduct vector surveillance?" An example spreadsheet, updated regularly and freely available, uses near real-time and forecast temperature data to generate guidance, based on a novel four level Zika risk code, for 733 US military facilities in the 50 states, the District of Columbia, and the territories of Guam and Puerto Rico.


Adaptation and validation of standardized aphasia tests in different languages: Lessons from the Boston Diagnostic Aphasia Examination - Short Form in Greek.
The aim of the current study was to adapt the Boston Diagnostic Aphasia Examination - Short Form (BDAE-SF) [1] to the Greek language and culture, determine the influence of demographic variables on performance and in particular the effects of age and education, develop normative data, and examine the discriminative validity of the test for acute stroke patients. A sample of 129 community healthy adults participated in the study (66 women), covering a broad range of ages and education levels so as to maximize representation of the Greek population and be able to examine the effects of age and education in language performance. Regression models showed that, overall, younger and more educated individuals presented higher performance on several subtests. Normative data for the Greek population are presented in percentile tables. Neurological patients' performance was compared to that of the neurologically intact population using Wilcoxon's rank sum test and for the most part was found to be significantly inferior, indicating good discriminant validity of the test. Qualitative errors of patients diagnosed with aphasia on the test are presented, and limitations and generalizable strengths of this adaptation are discussed.


Nodding syndrome: origins and natural history of a longstanding epileptic disorder in sub-Saharan Africa.
Repetitive involuntary head nodding was first reported in the 1960s in the Wapogoro tribe of Tanzania. We describe the natural history of head nodding in the Wapogoro tribe, with special reference to the earliest reported dates of onset. We analyzed clinical data from 150 historical patients seen between 1960 and 1971. Head nodding with or without grand mal convulsions was present in 33/150 (∼20%) cases, was mostly familial and equally distributed by gender. Age at onset of head nodding ranged from 2-22 years (mean: ∼10 years) in the period 1934-1962. Head nodding preceded onset of grand mal convulsions by up to 12 months, and motor and psychomotor deficits indicative of brain damage developed with time. Fourteen of the 33 cases died at 13-39 years of age (mean: ∼20 years) while nineteen aged 16-28 years (mean: ∼16 years) were still alive. Historical accounts of head nodding (amesinzia kichwa, Swahili) among the Wapogoro tribe fit the August 2012 World Health Organization (WHO) case definition of probable Nodding Syndrome. Reported to have existed in this population for at least 80 years, Nodding Syndrome is a progressive seizure disorder that leads to generalized convulsions (kifafa), brain damage and death.


Application of cost-effectiveness analysis to multiple products: a practical guide.
The appropriate interpretation of cost-effectiveness results and its use in treatment decision making have been debated vigorously. In this report we have summarized the decision rules described in the health economics literature to determine which intervention should be chosen from among multiple treatment options for a given disease, using a graphic framework. The implications of different means of expressing budget constraints on treatment choice are examined.


Diagnostic trends of disabled social security beneficiaries, 1986-93.
Growth in the number of applications and subsequent awards for Social Security disabled-worker benefits marked the period from 1986 through 1993. These increases resulted in the 37 percent rise in the number of disabled-worker beneficiaries, 6 out of 10 of whom had disabilities within three diagnostic groups: circulatory disorders; mental disorders (other than mental retardation); and musculoskeletal diseases. The percentage of disabled workers with a circulatory condition decreased from 21 to 14 percent, while the percentage with a mental disorder increased from 20 to 25 percent, and the percentage with musculoskeletal conditions increased from 18 to 21 percent. Musculoskeletal conditions (22 percent) were the leading diagnosis among disabled widows and widowers in 1993, while the disabled adult child population was dominated by the mental retardation diagnostic group (63 percent). Variations in diagnostic conditions of disabled workers by sex, age, and region were often substantial.


Are protected areas good for the human species? Effects of protected areas on rural depopulation in Spain.
Protected areas (PAs) seek to conserve valuable genes, species and ecosystems by applying a legal regime that restricts some socioeconomic activities and also offers opportunities for new ones. As a result, PAs have been claimed by some authors to boost socioeconomic conditions in rural areas mainly through tourism activities. However, others have claimed that PAs contribute to rural depopulation through the worsening of living conditions of local residents because of restrictions resulting from protection regulations. Here, we applied a multiple-paired Before-After-Control-Impact (BACI) research design on a census on protected rural municipalities (cases; N = 52) versus unprotected rural municipalities (controls; N = 55) in Spain to ascertain whether PAs had positive or negative effects on rural populations using three indicators on depopulation with official municipal data from 1996 until 2019: Compound annual growth rate (CAGR); Proportion of reproductive individuals (REP); and Proportion of reproductive females (WREP). We controlled for some confounders such as biophysical characteristics and regional regulations by carefully selecting our sample of municipalities spatially. Our results show that depopulation figures were worse in cases than in controls, with some exceptions whose characteristics should be further explored. Municipalities in Sites of Community Importance (SCIs) performed best against rural depopulation and generally better than their controls, whereas municipalities in Biosphere Reserves and Special Protection Areas (SPAs) showed mostly worse figures. Our findings suggest that, while necessary and important for biodiversity, multiple-use PAs generally entailed negative consequences for Spanish rural populations that need to be offset by State's intervention.