Thermal stress and toxicity.
Elevating ambient temperature above thermoneutrality exacerbates toxicity of most air pollutants, insecticides, and other toxic chemicals. On the other hand, safety and toxicity testing of toxicants and drugs is usually performed in mice and rats maintained at sub-thermoneutral temperatures of ~22∘C. When exposed to chemical toxicants under these relatively cool conditions, rodents typically undergo a regulated hypothermic response, characterized by preference for cooler ambient temperatures and controlled reduction in core temperature. Reducing core temperature delays the clearance of most toxicants from the body; however, a mild hypothermia also improves recovery and survival from the toxicant. Raising ambient temperature to thermoneutrality and above increases the rate of clearance of the toxicant but also exacerbates toxicity. Furthermore, heat stress combined with work or exercise is likely to worsen toxicity. Body temperature of large mammals, including humans, does not decrease as much in response to exposure to a toxicant. However, heat stress can nonetheless worsen toxic outcome in humans through a variety of mechanisms. For example, heat-induced sweating and elevation in skin blood flow accelerates uptake of some insecticides. Epidemiological studies suggest that thermal stress may exacerbate the toxicity of airborne pollutants such as ozone and particulate matter. Overall, translating results of studies in rodents to that of humans is a formidable task attributed in part to the interspecies differences in thermoregulatory response to the toxicants and to thermal stress.


Influence of nonfatal hospitalization for heart failure on subsequent mortality in patients with chronic heart failure.
Patients with chronic heart failure (HF) are at increased risk of both fatal and nonfatal major adverse cardiovascular events. We used data from the Candesartan in Heart failure: Assessment of Reduction in Mortality and morbidity (CHARM) trials to assess the influence of nonfatal hospitalizations for HF on subsequent mortality rates in a broad spectrum of HF patients. In the present study, 7599 patients with New York Heart Association class II to IV HF and reduced or preserved left ventricular ejection fraction were randomized to placebo or candesartan. We assessed the risk of death after discharge from a first hospitalization for HF using time-updated Cox proportional-hazards models on 7572 patients for whom discharge data were available. Of 7572 patients, 1455 (19%) had at least 1 HF hospitalization, and 586 of 1819 deaths occurred after discharge from an HF hospitalization. The mortality rate was increased after HF hospitalizations, even after adjustment for baseline predictors of death (hazard ratio, 3.15; 95% confidence interval, 2.83 to 3.50). Longer duration of HF hospitalization enhanced the risk of dying, as did repeat HF hospitalizations. Moreover, risk of death was highest within a month of discharge and then declined progressively over time, particularly for death resulting from HF progression and for sudden cardiac death. We observed a similar pattern of risk associated with all-cause hospitalization, although the magnitude was less than that with HF hospitalization. In patients with chronic HF, the risk of death is greatest in the early period after discharge after a hospitalization for HF and is directly related to the duration and frequency of HF hospitalizations. These findings suggest a role for increased surveillance in the early postdischarge period of greatest vulnerability after an HF admission.


Healthcare benefits linked with Below Poverty Line registration in India: Observations from Maharashtra Anaemia Study (MAS).
A 2015 Lancet paper by Patel et al. on healthcare access in India comprehensively discussed national health programmes where some benefits are linked with the country's Below Poverty Line (BPL) registration scheme. BPL registration aims to support poor families by providing free/subsidised healthcare. Technical issues in obtaining BPL registration by poor families have been previously reported in the Indian literature; however there are no data on family assets of BPL registrants. Here, we provide evidence of family-level assets among BPL registration holders (and non-BPL households) using original research data from the Maharashtra Anaemia Study (MAS). Social and health data from 287 pregnant women and 891 adolescent girls (representing 1178 family households) across 34 villages in Maharashtra state, India, were analysed. Several assets were shown to be similarly distributed between BPL and non-BPL households; a large proportion of families who would probably be eligible were not registered, whereas BPL-registered families often had significant assets that should not make them eligible. This is likely to be the first published evidence where asset distribution such as agricultural land, housing structures and livestock are compared between BPL and non-BPL households in a rural population. These findings may help planning BPL administration to allocate health benefits equitably, which is an integral part of national health programmes.


Perinatal hypophosphatasia: radiology, pathology and molecular biology studies in a family harboring a splicing mutation (648+1A) and a novel missense mutation (N400S) in the tissue-nonspecific alkaline phosphatase (TNSALP) gene.
We report on a postmortem diagnosis of perinatal lethal hypophosphatasia, an inborn error of metabolism characterized by a liver/bone/kidney alkaline phosphatase (ALP)-related defective bone mineralization due to mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene. Radiological and pathological studies identified a perinatal lethal hypophosphatasia showing a generalized bone mineralization defect including asymmetry of the cervical vertebral arches in a 22 +4 weeks' gestation fetus. Both parents revealed low serum ALP activities supporting the diagnosis. Sequencing analysis of the TNSALP gene showed two heterozygous mutations, 648+1A, a mutation affecting the donor splice site in exon 6, and N400S, a novel missense mutation in exon 11, located near the active site and very close to histidins 364 and 437, two crucial residues of the active site. Sequencing of exons 6 and 11 in the parents showed that 648+1A was from maternal origin and N400S from paternal origin. DNA-based prenatal testing in the subsequent pregnancy following a chorionic villous sampling performed at 10 weeks of gestation showed no mutation and a healthy infant was born at term.


Sex-specific and blood meal-induced proteins of Anopheles gambiae midguts: analysis by two-dimensional gel electrophoresis.
Anopheles gambiae is the main vector of Plasmodium falciparum in Africa. The mosquito midgut constitutes a barrier that the parasite must cross if it is to develop and be transmitted. Despite the central role of the mosquito midgut in the host/parasite interaction, little is known about its protein composition. Characterisation of An. gambiae midgut proteins may identify the proteins that render An. gambiae receptive to the malaria parasite. We carried out two-dimensional gel electrophoresis of An. gambiae midgut proteins and compared protein profiles for midguts from males, sugar-fed females and females fed on human blood. Very few differences were detected between male and female mosquitoes for the approximately 375 silver-stained proteins. Male midguts contained ten proteins not detected in sugar-fed or blood-fed females, which are therefore probably involved in male-specific functions; conversely, female midguts contained twenty-three proteins absent from male midguts. Eight of these proteins were specific to sugar-fed females, and another ten, to blood-fed females. Mass spectrometry analysis of the proteins found only in blood-fed female midguts, together with data from the recent sequencing of the An. gambiae genome, should make it possible to determine the role of these proteins in blood digestion or parasite receptivity.


RBCK1, an E3 ubiquitin ligase, interacts with and ubiquinates the human pregnane X receptor.
The pregnane X receptor (PXR, NR1I2) plays a pivotal role in the disposition and detoxification of numerous foreign and endogenous chemicals by increasing transcription of numerous target genes, including phase I and II drug-metabolizing enzymes and transporters. In the present study, yeast two-hybrid screening identified an E3 ubiquitin ligase, RBCK1 (Ring-B-box-coiled-coil protein interacting with protein kinase C-1), as a human pregnane X receptor (hPXR)-interacting protein. Coimmunoprecipitation studies confirmed the interaction between RBCK1 and hPXR when both were ectopically expressed in AD-293 cells. Domain mapping studies showed that the interaction between RBCK1 and hPXR involves all RBCK1 domains. We further demonstrate that RBCK1 ubiquitinates hPXR, and this may target hPXR for degradation by the ubiquitin-proteasome pathway. Simultaneous ectopic overexpression of RBCK1 and PXR decreased PXR levels in AD-293 cells, and this decrease was inhibited by the proteasomal inhibitor MG-132 (carbobenzoxy-Leu-Leu-leucinal). Furthermore, overexpression of RBCK1 decreased endogenous levels of PXR in HepG2 cells. Of importance, ectopic overexpression and silencing of endogenous RBCK1 in primary human hepatocytes resulted in a decrease and increase, respectively, in endogenous PXR protein levels and in the induction of PXR target genes by rifampicin. These results suggest that RBCK1 is important for the ubiquitination of PXR and may play a role in its proteasomal degradation.


Regenerative and non-regenerative calcium transients in hamster eggs triggered by inositol 1,4,5-trisphosphate.
1. Inositol 1,4,5-trisphosphate (InsP3) injected into unfertilized golden hamster eggs elicits a hyperpolarizing response (HR) that is due to stimulation of calcium-activated potassium channels in the egg plasma membrane. 2. A single injection of InsP3 gave a single HR above a threshold value of 0.3 nM. At 5 nM and above, InsP3 induced HRs with no detectable latency. At concentrations between these two values a latency was observed. The amplitude of the HR was independent of InsP3 concentration. 3. A second HR could be elicited by injection of InsP3, but five times more InsP3 was required to trigger a second HR, and 10-100 times more to give an HR of similar magnitude to the first, and there was no latency. 4. The increase in [Ca2+]i in response to an initial injection of 1 nM InsP3 could be resolved into two distinct components: a slow, early rise immediately after InsP3 injection (phase I) followed by a larger and more rapid increase (phase II). The initiation of an HR coincided with the second component of the [Ca2+]i increase. 5. Further injection of InsP3 resulted only in slow, smaller increases in [Ca2+]i that resembled phase I and often did not cause an HR. Phase II appeared to be absent. However, 100-fold greater InsP3 concentrations gave slow, larger Ca2+ transients (and HRs) with no detectable latency. 6. If large amounts of InsP3 were allowed to leak into the eggs constantly from a pipette, repetitive calcium transients were seen. Unlike the sustained repetitive responses seen at fertilization, they were often smaller than the initial transient and less well sustained. However, a subsequent transient could still be elicited on injection of very large concentrations of InsP3. 7. InsP3 can induce regenerative, all-or-none [Ca2+]i increase (CICR) in hamster eggs, often with a long latency, as well as non-regenerative increases. InsP3 injections desensitize CICR and cannot mimic all the features of Ca2+ signalling at fertilization in the hamster egg, in particular, the sensitization of CICR caused by the sperm.


Expression of the oncofetal protein IGF2BP3 in endometrial clear cell carcinoma: assessment of frequency and significance.
Insulin-like growth factor-II messenger RNA-binding protein 3 (IGF2BP3 or IMP3) is a biomarker whose expression has been found to be a negative prognostic factor in several neoplasms including ovarian clear cell carcinoma (CCC). In this study, we analyzed the frequency and clinicopathologic significance of IMP3 expression, as assessed by immunohistochemistry and as scored using a modified H-score system, in a cohort of 50 endometrial CCCs. Cases with scores of 0 to 100, 101 to 200, and 201 to 300 were classified as negative/mildly positive (n = 17), moderately positive (n = 20), and strongly positive (n = 13), respectively. A distinctive pattern of increased staining at the myoinvasive front (relative to the main tumor) was evident in 46% of the cases with evaluable foci of myometrial invasion. Moderate/strong IMP3 staining was associated with a tumor architectural pattern that has been reported to be of poor prognostic significance: at least 10% of the tumor composed of solid architecture or individual infiltrating tumor cells (P = .01). Increasing levels of IMP3 expression showed a trend toward decreasing relapse-free survival (RFS; median survival, 75.6, 81.3, and 48.4 months for the negative/mildly, moderately, and strongly positive groups, respectively [P = .09]). However, IMP3 expression was not significantly associated with reduced overall survival or RFS in a multivariate analytic model. The finding in a subset of our cases of increased IMP3 expression at the tumoral myoinvasive front is consistent with a role for IMP3 in invasiveness, as is the trend toward reduced RFS in cases expressing IMP3 at high levels. These preliminary findings suggest that IMP3 expression may be involved in the pathogenesis of CCC and is worthy of further exploration.


How state taxes and policies targeting soda consumption modify the association between school vending machines and student dietary behaviors: a cross-sectional analysis.
Sodas are widely sold in vending machines and other school venues in the United States, particularly in high school. Research suggests that policy changes have reduced soda access, but the impact of reduced access on consumption is unclear. This study was designed to identify student, environmental, or policy characteristics that modify the associations between school vending machines and student dietary behaviors. Data on school vending machine access and student diet were obtained as part of the National Youth Physical Activity and Nutrition Study (NYPANS) and linked to state-level data on soda taxes, restaurant taxes, and state laws governing the sale of soda in schools. Regression models were used to: 1) estimate associations between vending machine access and soda consumption, fast food consumption, and lunch source, and 2) determine if associations were modified by state soda taxes, restaurant taxes, laws banning in-school soda sales, or student characteristics (race/ethnicity, sex, home food access, weight loss behaviors.). Contrary to the hypothesis, students tended to consume 0.53 fewer servings of soda/week (95% CI: -1.17, 0.11) and consume fast food on 0.24 fewer days/week (95% CI: -0.44, -0.05) if they had in-school access to vending machines. They were also less likely to consume soda daily (23.9% vs. 27.9%, average difference = -4.02, 95% CI: -7.28, -0.76). However, these inverse associations were observed primarily among states with lower soda and restaurant tax rates (relative to general food tax rates) and states that did not ban in-school soda sales. Associations did not vary by any student characteristics except for weight loss behaviors. Isolated changes to the school food environment may have unintended consequences unless policymakers incorporate other initiatives designed to discourage overall soda consumption.


Digital ulcers predict a worse disease course in patients with systemic sclerosis.
Systemic sclerosis (SSc) is a systemic autoimmune disease with high morbidity and significant mortality. There is a great need of predictors that would allow risk stratification of patients with SSc and ultimately initiation of treatment early enough to ensure optimal clinical results. In this study, we evaluated whether a history of digital ulcers (HDU) at presentation may be a predictor of vascular outcomes and of overall clinical worsening and death in patients with SSc. Patients from the EULAR Scleroderma Trials and Research (EUSTAR) database, satisfying at inclusion the 1980 American College of Rheumatology classification criteria for SSc, who had a follow-up of at least 3 years since baseline or who have died, were included in the analysis. HDU at presentation as a predictor of disease worsening or death was evaluated by Cox proportional hazards regression analysis. 3196 patients matched the inclusion criteria (male sex 13.2%, 33.4% diffuse subset). At presentation, 1092/3196 patients had an HDU (34.1%). In multivariable analysis adjusting for age, gender and all parameters considered potentially significant, HDU was predictive for the presence of active digital ulcers (DUs) at prospective visits (HR (95% CI)): 2.41 (1.91 to 3.03), p<0.001, for an elevated systolic pulmonary arterial pressure on heart ultrasound (US-PAPs):1.36 (1.03 to 1.80), p=0.032, for any cardiovascular event (new DUs, elevated US-PAPs or LV failure): 3.56 (2.26 to 5.62), p<0.001, and for death (1.53 (1.16 to 2.02), p=0.003). In patients with SSc, HDU at presentation predicts the occurrence of DUs at follow-up and is associated with cardiovascular worsening and decreased survival.


Microstructure of ionic liquid (EAN)-rich and oil-rich microemulsions studied by SANS.
In a previous study we investigated the phase behavior of microemulsions consisting of the ionic liquid ethylammonium nitrate (EAN), an n-alkane and a nonionic alkyl polyglycolether (CiEj). We found the same general trends as for the aqueous counterparts, i.e. a transition from an oil-in-EAN microemulsion via a bicontinuous microemulsion to an EAN-in-oil microemulsion with increasing temperature. However, unlike what happens in the corresponding aqueous systems, in EAN-in-oil microemulsions only a very small amount of EAN was detected by NMR-measurements. This is why we investigated the phase behavior and microstructure of EAN-rich n-dodecane-in-EAN microemulsions and oil-rich EAN-in-n-octane microemulsions. We found that the ionic liquid emulsification failure boundary has an extraordinarily small slope, which suggests that the amphiphilic film loses its ability to solubilize EAN with an increase in temperature by only a few degrees. The analysis of the small angle neutron scattering (SANS) curves unambiguously shows that this behavior is due to the fact that the EAN molecules form a substructure with a characteristic length scale of Λ ≈ 8 Å inside the EAN-in-oil droplets. In more detail, the analysis of the SANS data with the GIFT method revealed a transition from spherical to cylindrical structures approaching the respective critical endpoint temperatures. By using the respective form factors and combining them with a Gaussian spatial intensity distribution to account for the EAN sub-structure we were able to describe the scattering curves nearly quantitatively.


A Multidisciplinary Biospecimen Bank of Renal Cell Carcinomas Compatible with Discovery Platforms at Mayo Clinic, Scottsdale, Arizona.
To address the need to study frozen clinical specimens using next-generation RNA, DNA, chromatin immunoprecipitation (ChIP) sequencing and protein analyses, we developed a biobank work flow to prospectively collect biospecimens from patients with renal cell carcinoma (RCC). We describe our standard operating procedures and work flow to annotate pathologic results and clinical outcomes. We report quality control outcomes and nucleic acid yields of our RCC submissions (N=16) to The Cancer Genome Atlas (TCGA) project, as well as newer discovery platforms, by describing mass spectrometry analysis of albumin oxidation in plasma and 6 ChIP sequencing libraries generated from nephrectomy specimens after histone H3 lysine 36 trimethylation (H3K36me3) immunoprecipitation. From June 1, 2010, through January 1, 2013, we enrolled 328 patients with RCC. Our mean (SD) TCGA RNA integrity numbers (RINs) were 8.1 (0.8) for papillary RCC, with a 12.5% overall rate of sample disqualification for RIN <7. Banked plasma had significantly less albumin oxidation (by mass spectrometry analysis) than plasma kept at 25 °C (P<.001). For ChIP sequencing, the FastQC score for average read quality was at least 30 for 91% to 95% of paired-end reads. In parallel, we analyzed frozen tissue by RNA sequencing; after genome alignment, only 0.2% to 0.4% of total reads failed the default quality check steps of Bowtie2, which was comparable to the disqualification ratio (0.1%) of the 786-O RCC cell line that was prepared under optimal RNA isolation conditions. The overall correlation coefficients for gene expression between Mayo Clinic vs TCGA tissues ranged from 0.75 to 0.82. These data support the generation of high-quality nucleic acids for genomic analyses from banked RCC. Importantly, the protocol does not interfere with routine clinical care. Collections over defined time points during disease treatment further enhance collaborative efforts to integrate genomic information with outcomes.


DNA methylation dynamics during intestinal stem cell differentiation reveals enhancers driving gene expression in the villus.
DNA methylation is of pivotal importance during development. Previous genome-wide studies identified numerous differentially methylated regions upon differentiation of stem cells, many of them associated with transcriptional start sites. We present the first genome-wide, single-base-resolution view into DNA methylation dynamics during differentiation of a mammalian epithelial stem cell: the mouse small intestinal Lgr5+ stem cell. Very little change was observed at transcriptional start sites and our data suggest that differentiation-related genes are already primed for expression in the stem cell. Genome-wide, only 50 differentially methylated regions were identified. Almost all of these loci represent enhancers driving gene expression in the differentiated part of the small intestine. Finally, we show that binding of the transcription factor Tcf4 correlates with hypo-methylation and demonstrate that Tcf4 is one of the factors contributing to formation of differentially methylated regions. Our results reveal limited DNA methylation dynamics during small intestine stem cell differentiation and an impact of transcription factor binding on shaping the DNA methylation landscape during differentiation of stem cells in vivo.


Signal integration at the level of ion channel and exocytotic function in pancreatic β-cells.
Whole body energy balance is ensured by the exquisite control of insulin secretion, the dysregulation of which has serious consequences. Although a great deal has been learned about the control of insulin secretion from pancreatic β-cells in the past 30 years, there remains much to be understood about the molecular mechanisms and interactions that underlie the precise control of this process. Numerous molecular interactions at the plasma membrane mediate the excitatory and amplifying events involved in insulin secretion; this includes interactions between ion channels, signal transduction machinery, and exocytotic proteins. The present Perspectives article considers evidence that key membrane and membrane-associated proteins essential to insulin secretion are regulated in concert as a functional unit, ensuring an integrated excitatory and exocytotic response to the signals that control insulin secretion.


A Dietary Supplement Containing Fucoidan Preserves Endothelial Glycocalyx through ERK/MAPK Signaling and Protects against Damage Induced by CKD Serum.
(1) Damage to the endothelial glycocalyx (eGC), a protective layer lining the endothelial luminal surface, is associated with chronic kidney disease (CKD), which leads to a worsening of cardiovascular outcomes in these patients. Currently, there are no targeted therapeutic approaches. Whether the dietary supplement EndocalyxTM (ECX) protects against endothelial damage caused by uremic toxins is unknown. (2) We addressed this question by performing atomic force microscopy measurements on living endothelial cells. We examined the effect of ECX on eGC thickness at baseline and with pooled serum from hemodialysis patients. ECX was also successfully administered in vivo in mice, in which eGC was assessed using perfused boundary region measurements by intravital microscopy of cremasteric vessels. (3) Both ECX and fucoidan significantly improved baseline eGC thickness. Our data indicate that these effects are dependent on ERK/MAPK and PI3K signaling. After incubation with eGC damaging serum from dialysis patients, ECX increased eGC height. Intravital microscopy in mice revealed a relevant increase in baseline eGC dimensions after feeding with ECX. (4) We identified a dietary supplement containing glycocalyx substrates and fucoidan as potential mediators of eGC preservation in vitro and in vivo. Our findings suggest that fucoidan may be an essential component responsible for protecting the eGC in acute settings. Moreover, ECX might contribute to both protection and rebuilding of the eGC in the context of CKD.


High subretinal fluid procoagulant activity in rhegmatogenous retinal detachment.
An increased mRNA expression of genes related to blood coagulation has been demonstrated in an experimental retinal detachment model but has not yet been confirmed in human clinical specimens. Tissue factor (TF), the initiating factor of blood coagulation, may be a determinant of the extent of tissue injury after rhegmatogenous retinal detachment (RRD). This study was conducted to determine whether subretinal fluid and vitreous fluid collected from patients with RRD have a procoagulant effect. Calibrated thrombin generation (CAT) was used to investigate the thrombogenic properties of 28 subretinal fluids collected during scleral buckling surgery for RRD. Further, the thrombogenic properties of vitreous fluids from RRD (n = 12), macular pucker (n = 5), macular hole (n = 6), and proliferative diabetic retinopathy (n = 5) were compared with the properties of eye bank eyes (n = 11), which served as control specimens. The procoagulant activity of TF was determined with Western blot analysis. The addition of subretinal fluid from all RRD patients (28/28, 100%) induced thrombin generation in normal and severely factor (F)XII-deficient plasma. Contrary to the subretinal fluid, the addition of vitreous fluids from various ocular disorders evoked very little thrombin generation in normal and severely FXII-deficient plasma (4/12, 33% RRD; 1/5, 20% macular pucker; 0/6, 0% macular hole; 0/5, 0% proliferative diabetic retinopathy; and 2/11, 18% eye bank eyes). The procoagulant activity in subretinal fluid was almost completely neutralized by antibodies against human TF. The presence of TF in subretinal fluid was confirmed by Western blot. Subretinal fluid of patients with RRD induces high procoagulant activity, determined by measuring the level of tissue factor.


Development of a biologically relevant dermal sampler.
There are currently no appropriate methods for measuring dermal exposure to volatile agents. To address this we have produced a prototype Institute of Occupational Medicine (IOM) dermal sampler consisting of an adsorbent sandwiched between a permeable membrane and an impervious backing. The concentration of solvent on the membrane surface may be estimated from the mass collected on the adsorbent and the known permeation rate through the membrane. We have developed the prototype IOM dermal sampler for measurement of toluene exposures. Evaluation of the prototype sampler was undertaken in two stages: laboratory performance in controlled exposure situations and two short-field evaluations, which included simultaneous measurement of inhalation exposure. In all cases we compared the prototype IOM dermal sampler with activated charcoal cloth (ACC). Laboratory trials were split into spray, pour and immersion tests. The data from these suggest that the sampler responds to concentration rather than the mass on the surface of the sampler. The field study showed that the prototype sampler was suitable for measuring dermal exposure. However, the mean permeation rate of the best membrane was 78 000 mug cm(-2) h(-1), which is higher than the permeation rate through skin. This high permeation rate created difficulties throughout the study, particularly as it allowed the adsorbent to become rapidly saturated. The prototype IOM dermal sampler is the first practical dermal exposure sampler to mimic uptake through the skin. The sampler gave reproducible results in the laboratory and field trials. Future work is required to identify a less permeable membrane, which has characteristics closer to that of human skin. Additionally, a higher capacity adsorbent would be desirable. We have demonstrated a major difference when calculating the total contribution to body burden via the dermal exposure pathway using the prototype IOM dermal sampler and ACC patches, 1.5% of the total body burden compared with 95%. The prototype IOM dermal sampler provides a more biologically relevant exposure metric than the alternatives.


STAT3 interrupts ATR-Chk1 signaling to allow oncovirus-mediated cell proliferation.
DNA damage response (DDR) is a signaling network that senses DNA damage and activates response pathways to coordinate cell-cycle progression and DNA repair. Thus, DDR is critical for maintenance of genome stability, and presents a powerful defense against tumorigenesis. Therefore, to drive cell-proliferation and transformation, viral and cellular oncogenes need to circumvent DDR-induced cell-cycle checkpoints. Unlike in hereditary cancers, mechanisms that attenuate DDR and disrupt cell-cycle checkpoints in sporadic cancers are not well understood. Using Epstein-Barr virus (EBV) as a source of oncogenes, we have previously shown that EBV-driven cell proliferation requires the cellular transcription factor STAT3. EBV infection is rapidly followed by activation and increased expression of STAT3, which mediates relaxation of the intra-S phase cell-cycle checkpoint; this facilitates viral oncogene-driven cell proliferation. We now show that replication stress-associated DNA damage, which results from EBV infection, is detected by DDR. However, signaling downstream of ATR is impaired by STAT3, leading to relaxation of the intra-S phase checkpoint. We find that STAT3 interrupts ATR-to-Chk1 signaling by promoting loss of Claspin, a protein that assists ATR to phosphorylate Chk1. This loss of Claspin which ultimately facilitates cell proliferation is mediated by caspase 7, a protein that typically promotes cell death. Our findings demonstrate how STAT3, which is constitutively active in many human cancers, suppresses DDR, fundamental to tumorigenesis. This newly recognized role for STAT3 in attenuation of DDR, discovered in the context of EBV infection, is of broad interest as the biology of cell proliferation is central to both health and disease.


Y chromosome haplotype distribution of brown bears (Ursus arctos) in Northern Europe provides insight into population history and recovery.
High-resolution, male-inherited Y-chromosomal markers are a useful tool for population genetic analyses of wildlife species, but to date have only been applied in this context to relatively few species besides humans. Using nine Y-chromosomal STRs and three Y-chromosomal single nucleotide polymorphism markers (Y-SNPs), we studied whether male gene flow was important for the recent recovery of the brown bear (Ursus arctos) in Northern Europe, where the species declined dramatically in numbers and geographical distribution during the last centuries but is expanding now. We found 36 haplotypes in 443 male extant brown bears from Sweden, Norway, Finland and northwestern Russia. In 14 individuals from southern Norway from 1780 to 1920, we found two Y chromosome haplotypes present in the extant population as well as four Y chromosome haplotypes not present among the modern samples. Our results suggested major differences in genetic connectivity, diversity and structure between the eastern and the western populations in Northern Europe. In the west, our results indicated that the recovered population originated from only four male lineages, displaying pronounced spatial structuring suggestive of large-scale population size increase under limited male gene flow within the western subpopulation. In the east, we found a contrasting pattern, with high haplotype diversity and admixture. This first population genetic analysis of male brown bears shows conclusively that male gene flow was not the main force of population recovery.


Electron density-based transformation of trimethoprim during biological wastewater treatment.
This research investigated the biological transformation of trimethoprim (TMP). Partial TMP removal was observed in the presence of ammonia and toluene, and increasing the solids retention time from 20 days to 60 days improved TMP removal in both the nitrifying and heterotrophic bioreactors. Two TMP-related metabolites were identified, the first (5-(3,4,5-trimethoxybenzyl) pyrimidine-2,4-diamine, 5-hydroxyl) showing that a hydroxylation reaction took place, and the second (5-(1-carboxyl, 1-methoxy, 5-methoxy 1-,4-pentene) pyrimidine-2,4-diamine, 5-hydroxyl) showing that the trimethoxybenzyl ring was cleaved. This research is the first that we are aware of to report these two TMP-related byproducts. TMP metabolites show that initiating reactions take place where the electron density is highest, and that these initiating reactions shift the electron density of TMP, likely affecting the course of transformation.


Physical Performance Assessments of Strength and Power in Women Collegiate Athletes.
Limited research exists on physical performance assessments for women collegiate athletes. The current cross-sectional study compared field-based tests of muscular strength and power and investigated their relationship. Sports included field hockey, volleyball, soccer, and softball. Tests of one repetition maximum (1-RM) back squat, 1-RM bench press, vertical jump, and standing long jump were administered. A one-way analysis of variance (ANOVA) assessed differences across sports. Bivariate Pearson correlation coefficients examined relationships among tests. It was hypothesized sports with a higher anaerobic nature (volleyball, softball) would outperform those with higher aerobic nature (field hockey, soccer). Softball had the highest 1-RM bench press and 1-RM back squat (p < 0.001) compared to field hockey, volleyball, and soccer. Further, softball had the highest vertical jump (p < 0.001) compared to field hockey and soccer, but did not differ from volleyball. There were no differences across sports for standing long jump. Correlations (p < 0.001) existed between 1-RM back squat and 1-RM bench press (n = 663, r = 0.56), and vertical jump and standing long jump (n = 160, r = 0.64). Results demonstrate strength and power differences among collegiate women's sports. Softball consistently outperformed others in bench press, back squat, and vertical jump, which may be due to the demand of power embedded in the nature of the sport. These data provide descriptive measures of physical performance assessments and may assist practitioners with goal setting and program design.


Cellular Therapies for Muscular Dystrophies: Frustrations and Clinical Successes.
Cell-based therapy for muscular dystrophies was initiated in humans after promising results obtained in murine models. Early trials failed to show substantial clinical benefit, sending researchers back to the bench, which led to the discovery of many hurdles as well as many new venues to optimize this therapeutic strategy. In this review we summarize progress in preclinical cell therapy approaches, with a special emphasis on human cells potentially attractive for human clinical trials. Future perspectives for cell therapy in skeletal muscle are discussed, including the perspective of combined therapeutic approaches.


The effect of flow rate and catheter diameter on hemolysis during blood transfusions using canine packed red blood cells.
To evaluate the degree of hemolysis in canine packed RBCs at varying catheter sizes and flow rates as determined by RBC count, Hct, hemoglobin, creatine phosphokinase, and phosphorus. This study hypothesized that changes in flow rate and catheter diameter would change the degree of hemolysis. A fresh unit of canine RBCs. A fresh unit of purchased canine packed RBCs was run through an IV infusion pump through 5 different catheter sizes (16, 18, 20, 22, and 24 gauge) at 5 different rates (50, 250, 500, 750, and 999 mL/h). Each sample was submitted for a CBC and chemistry, and RBC count, hemoglobin, hematocrit, creatine phosphokinase, and phosphorus were specifically evaluated to assess for the degree of hemolysis. Compared to the control, flow rate did not significantly affect the degree of hemolysis. Smaller catheter sizes had a significantly increased hemolysis (P < .05) based on variables to evaluate hemolysis (RBC count, Hct, hemoglobin, creatine phosphokinase, and phosphorus). This study's hypothesis was upheld in association with the catheter diameter but was rejected with flow rate. Blood transfusions are a common practice in veterinary medicine as treatment of anemia and hemorrhage. Hemolysis was greater when small catheter sizes were used. Larger catheters are recommended when feasible when performing transfusions.


A long, naturally presented immunodominant epitope from NY-ESO-1 tumor antigen: implications for cancer vaccine design.
The tumor antigen NY-ESO-1 is a promising cancer vaccine target. We describe here a novel HLA-B7-restricted NY-ESO-1 epitope, encompassing amino acids 60-72 (APRGPHGGAASGL), which is naturally presented by melanoma cells. The tumor epitope bound to HLA-B7 by bulging outward from the peptide-binding cleft. This bulged epitope was not an impediment to T-cell recognition, however, because four of six HLA-B7(+) melanoma patients vaccinated with NY-ESO-1 ISCOMATRIX vaccine generated a potent T-cell response to this determinant. Moreover, the response to this epitope was immunodominant in three of these patients and, unlike the T-cell responses to bulged HLA class I viral epitopes, the responding T cells possessed a remarkably broad TCR repertoire. Interestingly, HLA-B7(+) melanoma patients who did not receive the NY-ESO-1 ISCOMATRIX vaccine rarely generated a spontaneous T-cell response to this cryptic epitope, suggesting a lack of priming of such T cells in the natural anti-NY-ESO-1 response, which may be corrected by vaccination. Together, our results reveal several surprising aspects of antitumor immunity and have implications for cancer vaccine design.


Risk factors for osteoporotic fractures and low bone density in pre and postmenopausal women.
To estimate the prevalence and analyze risk factors associated to osteoporosis and low-trauma fracture in women. Cross-sectional study including a total of 4,332 women older than 40 attending primary care services in the Greater São Paulo, Southeastern Brazil, between 2004 and 2007. Anthropometrical and gynecological data and information about lifestyle habits, previous fracture, medical history, food intake and physical activity were obtained through individual quantitative interviews. Low-trauma fracture was defined as that resulting from a fall from standing height or less in individuals 50 years or older. Multiple logistic regression models were designed having osteoporotic fracture and bone mineral density (BMD) as the dependent variables and all other parameters as the independent ones. The significance level was set at p<0.05. The prevalence of osteoporosis and osteoporotic fractures was 33% and 11.5%, respectively. The main risk factors associated with low bone mass were age (OR=1.07; 95% CI: 1.06;1.08), time since menopause (OR=2.16; 95% CI: 1.49;3.14), previous fracture (OR=2.62; 95% CI: 2.08;3.29) and current smoking (OR=1.45; 95% CI: 1.13;1.85). BMI (OR=0.88; 95% CI: 0.86;0.89), regular physical activity (OR=0.78; 95% CI: 0.65;0.94) and hormone replacement therapy (OR=0.43; 95% CI: 0.33;0.56) had a protective effect on bone mass. Risk factors significantly associated with osteoporotic fractures were age (OR=1.05; 95% CI: 1.04;1.06), time since menopause (OR=4.12; 95% CI: 1.79;9.48), familial history of hip fracture (OR=3.59; 95% CI: 2.88;4.47) and low BMD (OR=2.28; 95% CI: 1.85;2.82). Advanced age, menopause, low-trauma fracture and current smoking are major risk factors associated with low BMD and osteoporotic fracture. The clinical use of these parameters to identify women at higher risk for fractures might be a reasonable strategy to improve the management of osteoporosis.


Replication-competent noninduced proviruses in the latent reservoir increase barrier to HIV-1 cure.
Antiretroviral therapy fails to cure HIV-1 infection because latent proviruses persist in resting CD4(+) T cells. T cell activation reverses latency, but <1% of proviruses are induced to release infectious virus after maximum in vitro activation. The noninduced proviruses are generally considered defective but have not been characterized. Analysis of 213 noninduced proviral clones from treated patients showed 88.3% with identifiable defects but 11.7% with intact genomes and normal long terminal repeat (LTR) function. Using direct sequencing and genome synthesis, we reconstructed full-length intact noninduced proviral clones and demonstrated growth kinetics comparable to reconstructed induced proviruses from the same patients. Noninduced proviruses have unmethylated promoters and are integrated into active transcription units. Thus, it cannot be excluded that they may become activated in vivo. The identification of replication-competent noninduced proviruses indicates that the size of the latent reservoir-and, hence, the barrier to cure-may be up to 60-fold greater than previously estimated.


Asynchronous Video Interviewing as a New Technology in Personnel Selection: The Applicant's Point of View.
The present study aimed to integrate findings from technology acceptance research with research on applicant reactions to new technology for the emerging selection procedure of asynchronous video interviewing. One hundred six volunteers experienced asynchronous video interviewing and filled out several questionnaires including one on the applicants' personalities. In line with previous technology acceptance research, the data revealed that perceived usefulness and perceived ease of use predicted attitudes toward asynchronous video interviewing. Furthermore, openness revealed to moderate the relation between perceived usefulness and attitudes toward this particular selection technology. No significant effects emerged for computer self-efficacy, job interview self-efficacy, extraversion, neuroticism, and conscientiousness. Theoretical and practical implications are discussed.


Evolutionary relationships of completely sequenced Clostridia species and close relatives.
The class Clostridia in the phylum Firmicutes includes a very heterogeneous assemblage of bacteria. Their evolutionary relationships are not well established; revisions of their phylogenetic placements based on comparative studies of 16S rRNA gene sequences are in progress as genome sequence information accumulates. In this work, phylogenetic trees were reconstructed based on 21 concatenated ribosomal protein sequences using Bayesian and maximum-likelihood methods. Both trees consistently indicate that the Halanaerobiales is a deeply branching order among the class Clostridia. The rest of the Clostridia species are grouped into 10 monophyletic clusters, most of which are comprised of two or three orders and families according to the current Clostridial taxonomy. The maximum-likelihood tree placed Coprothermobacter proteolyticus and Thermodesulfobium narugense in the class Clostridia in accordance with the current taxonomy, in which these two bacteria are assigned to the family Thermodesulfobiaceae. However, the Bayesian tree placed these two bacteria at the boundary between the Firmicutes and Actinobacteria. A gene arrangement that is present uniquely in the Firmicutes species was identified. Both Coprothermobacter proteolyticus and Thermodesulfobium narugense do not have this arrangement characteristic of the Firmicutes. On the basis of the Bayesian tree and gene arrangement comparison, it is suggested that Coprothermobacter proteolyticus and Thermodesulfobium narugense should be placed outside the phylum Firmicutes.


"We are the heroes because we are ready to die for this country": Participants' decision-making and grounded ethics in an Ebola vaccine clinical trial.
The 2014-2016 Ebola epidemic presented a challenging setting in which to carry out clinical trials. This paper reports findings from social science research carried out in Kambia, Northern Sierra Leone during first year of an Ebola vaccine trial (August 2015-July 2016). The social science team collected data through ethnographic observation, 42 in depth interviews; 4 life narratives; 200 exit interviews; 31 key informant interviews; and 8 focus group discussions with trial participants and community members not enrolled in the trial. Whilst research often focuses on why people refuse vaccination, we instead explore participant motivations for volunteering for the study, in spite of prevailing anxieties, rumours and mistrust during and after the Ebola outbreak. In so doing the paper contributes to on-going debates about research ethics and community engagement in resource poor contexts, offering reflections from an emergency and post-epidemic setting. We analyse participants' perceptions of the risks and benefits of participations, highlighting the importance of a contextual approach. We focus on four types of motivation: altruism; curiosity and hope; health-seeking; and notions of exchange, and argue for the role of social science in developing grounded research ethics and community engagement strategies that can take into account context and local realities.


Estimating the prevalence and burden of major disorders of the brain in Nepal: methodology of a nationwide population-based study.
The major disorders of the brain (MDBs), in terms of their prevalence and the burdens of ill health, disability and financial cost that they impose on individuals and society, are headache, depression and anxiety. No population-based studies have been conducted in Nepal. Our purpose was to assess the prevalence and burden attributable to MDBs in Nepal in order to inform health policy. Here we report the methodology. The unusual sociocultural diversity and extreme geographical variation of the country required adaptation of standard methodology. We ran pre-pilot and pilot studies before embarking on the main study. The study design was cross-sectional. The population of interest were adults aged 18-65 years who were Nepali speaking and living in Nepal. We selected, employed and trained groups of interviewers to visit randomly selected households by cold-calling. Households were selected from 15 representative districts out of 75 in the country through multistage cluster sampling. One participant was selected randomly from each household. We used structured questionnaires (the HARDSHIP questionnaire, Hospital Anxiety and Depression Scale, and Eysenck Personality Questionnaire -Neuroticism), culturally adapted and translated into Nepali. We recorded blood pressure, weight, height and waist circumference, and altitude of each household. We implemented various quality-assurances measures. We completed the survey in one month, prior to onset of the monsoon. Among 2,210 selected households, all were contacted, 2,109 were eligible for the study and, from these, 2,100 adults participated. The participation rate was 99.6%. Standard methodology was successfully applied in Nepal, with some adaptations. The sociocultural and extraordinary geographic diversity were challenging, but did not require us to compromise the scientific quality of the study.


Single-molecule junctions with epitaxial graphene nanoelectrodes.
On the way to ultraflat single-molecule junctions with transparent electrodes, we present a fabrication scheme based on epitaxial graphene nanoelectrodes. As a suitable molecule, we identified a molecular wire with fullerene anchor groups. With these two components, stable electrical characteristics could be recorded. Electrical measurements show that single-molecule junctions with graphene and with gold electrodes display a striking agreement. This motivated a hypothesis that the differential conductance spectra are rather insensitive to the electrode material. It is further corroborated by the assignment of asymmetries and spectral features to internal molecular degrees of freedom. The demonstrated open-access graphene electrodes and the electrode-insensitive molecules provide a model system that will allow for a thorough investigation of an individual single-molecule contact with additional probes.


Nutritional supplementation for Alzheimer's disease?
Evidence for the benefit of nutrition in Alzheimer's disease continues to accumulate. Many studies with individual vitamins or supplements show marginal, if any, benefit. However, new findings with combinatorial formulations demonstrate improvement in cognitive performance and behavioral difficulties that accompany Alzheimer's disease. Herein, we review some of the most recent clinical advances and summarize supportive preclinical studies. We present novel positive effects on Alzheimer's disease derived from diet, trace elements, vitamins and supplements. We discuss the inherent difficulty in conducting nutritional studies because of the variance in participants' nutritional history, versus pharmacological interventions in which participants are naive to the intervention. We examine the evidence that epigenetics play a role in Alzheimer's disease and how nutritional intervention can modify the key epigenetic events to maintain or improve cognitive performance. Overall consideration of the most recent collective evidence suggests that the optimal approach for Alzheimer's disease would seem to combine early, multicomponent nutritional approaches (a Mediterranean-style diet, multivitamins and key combinatorial supplements), along with lifestyle modifications such as social activity and mental and physical exercise, with ultimate addition of pharmacological agents when warranted.


FAST: A Novel, Executive Function-Based Approach to Cognitive Enhancement.
The present study introduces a novel cognitive intervention aimed at improving fluid intelligence (Gf), based on a framework we refer to as FAST: Flexible, Adaptive, Synergistic Training. FAST leverages a combination of novel game-based executive function (EF) training-designed specifically to enhance the likelihood of transfer-and transcranial electrical stimulation (tES), with aims to synergistically activate and strengthen mechanisms of cognitive control critical to Gf. To test our intervention, we collected three Gf measures from 113 participants [the advanced short Bochumer Matrizen-Test (BOMAT), Raven's Advanced Progressive Matrices (APM), and matrices similar to Raven's generated by Sandia labs], prior to and following one of three interventions: (1) the FAST + tRNS intervention, a combination of 30 min of daily training with our novel training game, Robot Factory, and 20 min of concurrent transcranial random noise stimulation applied to bilateral dorsolateral prefrontal cortex (DLPFC); (2) an adaptively difficult Active Control intervention comprised of visuospatial tasks that specifically do not target Gf; or (3) a no-contact control condition. Analyses of changes in a Gf factor from pre- to post-test found numerical increases for the FAST + tRNS group compared to the two control conditions, with a 0.3 SD increase relative to Active Control (p = 0.07), and a 0.19 SD increase relative to a No-contact control condition (p = 0.26). This increase was found to be largely driven by significant differences in pre- and post-test Gf as measured on the BOMAT test. Progression through the FAST training game (Robot Factory) was significantly correlated with changes in Gf. This is in contrast with progress in the Active Control condition, as well as with changes in individual EFs during FAST training, which did not significantly correlate with changes in Gf. Taken together, this research represents a useful step forward in providing new insights into, and new methods for studying, the nature of Gf and its malleability. Though our results await replication and extension, they provide preliminary evidence that the crucial characteristic of Gf may, in fact, be the ability to combine EFs rapidly and adaptively according to changing demand, and that Gf may be susceptible to targeted training.


Quarter-salt formation defining the anomalous temperature dependence of the aqueous solubility of sodium monododecyl phosphate.
The salts of monoalkyl phosphates (MAPs) have been identified as a class of inherently mild surfactants for use in household and personal products. They represent an anionic species intermediate in terms of pKa between the sulfates and the carboxylates and are analogous to the carboxylates in that they form acid-salts (which are here termed quarter-salts)-hydrogen-bonded dimers consisting of an undissociated MAP acid and an MAP monosalt. These complexes precipitate from solutions of the monosalt over a range of lower MAP concentrations giving rise to an unusual solubility/temperature relationship. The solubility of monosodium monododecyl phosphate (NaC(12)MAP) increases with temperature up to 0.01 M at approximately 60 degrees C, which corresponds to the conventional Krafft point as shown by the appearance of micelles in solution. The solubility then increases further to approximately 0.04 M as the solubility temperature declines from 60 to 38 degrees C. The transition between these two trends is characterized by a rather sharp temperature maximum in the solubility curve. In a third stage, the solubility then rises rapidly with very small change of temperature. This unusual overall behavior is shown to correspond with three distinct solid-phase compositions for the precipitates at temperatures below the solubility curve. At the lowest concentrations and up through the Krafft Point, the solid phase has been identified as the stoichiometric quarter-salt. Over the declining temperature portion of the solubility curve, the supernatant solution coexists with a macroscopic mixture of separate quarter-salt and monosalt solids. In the high-concentration third region the solid phase is exclusively the MAP monosalt. The coprecipitation of quarter-salt and monosalt from the monosalt solution occurs reversibly in the declining portion of the solubility curve and is accompanied by an increase in pH. The four phase system (solution, vapor, and two pure solid phases) retains one degree of freedom according to the phase rule since the system is in effect three component in that region.


Targeting myomiRs by tocotrienol-rich fraction to promote myoblast differentiation.
Several muscle-specific microRNAs (myomiRs) are differentially expressed during cellular senescence. However, the role of dietary compounds on myomiRs remains elusive. This study aimed to elucidate the modulatory role of tocotrienol-rich fraction (TRF) on myomiRs and myogenic genes during differentiation of human myoblasts. Young and senescent human skeletal muscle myoblasts (HSMM) were treated with 50 μg/mL TRF for 24 h before and after inducing differentiation. The fusion index and myotube surface area were higher (p < 0.05) on days 3 and 5 than that on day 1 of differentiation. Ageing reduced the differentiation rate, as observed by a decrease in both fusion index and myotube surface area in senescent cells (p < 0.05). Treatment with TRF significantly increased differentiation at days 1, 3 and 5 of young and senescent myoblasts. In senescent myoblasts, TRF increased the expression of miR-206 and miR-486 and decreased PTEN and PAX7 expression. However, the expression of IGF1R was upregulated during early differentiation and decreased at late differentiation when treated with TRF. In young myoblasts, TRF promoted differentiation by modulating the expression of miR-206, which resulted in the reduction of PAX7 expression and upregulation of IGF1R. TRF can potentially promote myoblast differentiation by modulating the expression of myomiRs, which regulate the expression of myogenic genes.


A Decade of Global mRNA and miRNA Profiling of HPV-Positive Cell Lines and Clinical Specimens.
For more than a decade, global gene expression profiling has been extensively used to elucidate the biology of human papillomaviruses (HPV) and their role in cervical- and head-and-neck cancers. Since 2008, the expression profiling of miRNAs has been reported in multiple HPV studies. Two major strategies have been employed in the gene and miRNA profiling studies: In the first approach, HPV positive tumors were compared to normal tissues or to HPV negative tumors. The second strategy relied on analysis of cell cultures transfected with single HPV oncogenes or with HPV genomes compared to untransfected cells considered as models for the development of premalignant and malignant transformations.In this review, we summarize what we have learned from a decade of global expression profiling studies. We performed comprehensive analysis of the overlap of the lists of differentially expressed genes and microRNAs, in both tissue samples and cell culture based studies. The review focuses mainly on HPV16, however reports from other HPV species are used as references. We discuss the low degree of consensus among different studies and the limitation of differential expression analysis as well as the fragmented miRNA-mRNA target correlation evidence. Furthermore, we propose an approach for future research to include more comprehensive miRNA-mRNA target correlation analysis and to apply systems biology/gene networks methodology.


FGFR1 fusion kinase regulation of MYC expression drives development of stem cell leukemia/lymphoma syndrome.
Oncogenic transformation of hematopoietic stem cells by chimeric fusion kinases causing constitutive activation of FGFR1 leads to a stem cell leukemia/lymphoma (SCLL) syndrome, accompanied by widespread dysregulation of gene activity. We now show that FGFR1 activation is associated with upregulation of MYC and pharmacological suppression of FGFR1 activation leads to downregulation of MYC and suppression of MYC target genes. Luciferase reporter assays demonstrate that FGFR1 can directly regulate MYC expression and this effect is enhanced in the presence of chimeric FGFR1 kinases. In SCLL cells, a truncated form of FGFR1 is generated by granzyme B cleavage of the chimeric kinases, producing a nucleus-restricted derivative that can bind MYC regulatory regions. Mutation of the granzyme B cleavage site prevents relocation to the nucleus but does not suppress MYC activation, suggesting additional mechanisms of MYC activation in the presence of cytoplasm-restricted chimeric kinases. We show that one of these mechanisms involves activating cytoplasmic STAT5, which upregulates MYC independent of the truncated FGFR1 kinase. Targeting MYC function using shRNA knockdown and 10054-F8 in SCLL cells leads to inhibition of cell proliferation and synergizes with the BGJ398 FGFR1 inhibitor, suggesting a combination therapy that could be used in the treatment of SCLL.


Addressing the socioeconomic determinants of adolescent health: experiences from the WHO/HBSC Forum 2007.
Over the past 25 years, the WHO collaborative cross-national Health Behaviour in School-aged Children (HBSC) study has been accumulating evidence that provides insights into how to promote the health and well-being of young people. HBSC has increased understanding of the determinants of young people's health, particularly in relation to the social contexts in which they live, learn and play. The study now spans 43 countries and regions in Europe and North America. HBSC provides intelligence for the development and evaluation of public health policy and practice at national, sub-national and international levels. However, the mere existence of evidence does not automatically change policy nor necessarily improve the lives of young people. Effective mechanisms to ensure use of evidence in policy-making and practice are needed. The WHO/HBSC Forum series is a platform designed to facilitate the translation of evidence into action. Forum processes convene researchers, policy-makers and practitioners from across Europe to analyse data, review policies and interventions, and identify lessons learned to improve the health of adolescents through actions that address the social contexts that influence their health. Each Forum process consists of case studies produced by interdisciplinary teams in countries and regions, cross-country evidence reviews, a European consultation, an outcomes statement within a final publication, and a Web-based knowledge platform. In addition to emphasizing the translation of research into action, the Forum series focuses on increasing know-how to scale up intersectoral policies and interventions; reduce health inequities; and involve young people in the design, implementation and evaluation of policies and interventions. Interviews with selected participants in the 2007 Forum process revealed that national-level impacts of involvement were: brokering new or strengthening existing working relationships among members of case study drafting teams and national delegations to events; feeding into the formulation of national policy or practice design; and enabling the comparison of information systems, policies, interventions, and working methods with other countries and against the evidence base, thus providing encouragement for new and/or validation of existing activities. The WHO/HBSC Forum series is an evolving platform. The methodology for each Forum process incorporates lessons learnt through past Fora - within resource constraints - and is based on the principles of efficiency and effectiveness. Areas requiring further development, identified through the aforementioned interviews and based on the reflections of co-organizers, include identification of means to ensure systematic, appropriate and meaningful youth involvement; maximization of the usefulness of the European consultation; and definition of a budget line and framework for evaluation of the process' impact at country level.


Magnification endoscopy in esophageal squamous cell carcinoma: a review of the intrapapillary capillary loop classification.
Recent developments in image-enhancement technology have enabled clear visualization of the microvascular structure of the esophageal mucosa. In particular, intrapapillary capillary loops (IPCLs) are observed as brown loops on magnification endoscopy with narrow-band imaging (NBI). IPCLs demonstrate characteristic morphological changes according to the structural irregularity of esophageal epithelium and cancer infiltration, summarized in the IPCL classification. In this review, the process from the first endoscopic description of IPCLs to the eventual development of the IPCL classification is described and discussed, particularly focusing on early stage squamous cell carcinoma of the esophagus.


Is 24 h ambulatory oesophageal pH monitoring useful in a district general hospital?
53 of 61 patients successfully completed 24 h ambulatory monitoring of oesophageal pH. The indications were: symptoms suggestive of gastro-oesophageal reflux but with normal endoscopy (19 cases); atypical chest pain with normal endoscopy (21 cases); or respiratory symptoms possibly due to reflux (13 cases). A temporal association between abnormal reflux and the presenting symptoms was demonstrated in 25 patients (47%). 6 patients were shown to have respiratory symptoms after episodes of reflux which resolved on treatment of the reflux alone. Reflux occurring only in the erect posture was observed in some patients and may have been a manifestation of the irritable bowel syndrome. Reflux as a cause of symptoms was excluded in 14 patients. The procedure was well tolerated in most patients, simple to operate, and inexpensive.


Voltage-gated and inwardly rectifying potassium channels.
This lecture is dedicated to Max Delbrück and Seymour Benzer. Max Delbrück was our graduate advisor. He introduced us to a variety of biophysical problems, and taught us ways of thinking about these problems by example. Potassium channels was one of the topics included in his journal club in the early seventies; Max also carefully considered the feasibility of purifying potassium channels then. It was in Seymour Benzer's laboratory that we began to look for Drosophila mutants that affect synaptic transmission at the larval neuromuscular junction. Shaker was the first behavioural mutant we tested that gave a robust phenotype, a phenotype that could be mimicked by treating wild-type preparations with a potassium channel blocker. This mutant fly has led us to our subsequent molecular studies of potassium channels. Since we settled in the University of California, San Francisco, and began to study neural development as well as potassium channels, we have settled into the pattern of each attending meetings and presenting our studies on one of these two areas so as to avoid both being away from home and our children at the same time. In following this pattern, I will be presenting the studies of potassium channels as part of our long-term collaboration. In this talk I will first briefly take you through the path that led us to the molecular studies of potassium channels and then discuss the diversity and modulation of these potassium channels at the molecular and physiological level.


Caregivers' Attitude and Perspective on Coercion and Restraint Practices on Psychiatric Inpatients from South India.
Coercion and restraint practices in psychiatric care are common phenomena and often controversial and debatable ethical issue. Caregivers' attitude and perspective on coercion and restraint practices on psychiatric inpatients have received relatively less research attention till date. Caregivers' attitude and perspective on coercion and restraint practices on psychiatric inpatients. This is a hospital-based, a descriptive, cross-sectional study. A total of 200 (n = 200) consecutive patient and their caregivers were chosen between June 2013 and September 2014 through computer-generated random numbers sampling technique. We used a semi-structured interview questionnaire to capture caregivers' attitude and perspective on coercion and restraint practices. Sociodemographic and coercion variable were analyzed using descriptive statistics. McNemar test was used to assess discrete variables. The mean age was 43.8 (±14.9) years. About 67.5% of the caregivers were family members, 60.5% of them were male and 69.5% were from low-socioeconomic status. Caregivers used multiple methods were used to bring patients into the hospital. Threat (52.5%) was the most common method of coercion followed by persuasion (48.5%). Caregivers felt necessary and acceptable to use chemical restraint (82.5%), followed by physical restraint (71%) and electroconvulsive therapy (ECT) (56.5%) during acute and emergency psychiatric care to control imminent risk behavior of patients. Threat, persuasion and physical restraint were the common methods to bring patients to bring acutely disturbed patients to mental health care. Most patients caregivers felt the use of chemical restraint, physical restraint and ECT as necessary for acute and emergency care in patients with mental illness.


Estrogen metabolism and risk of breast cancer: a prospective study of the 2:16alpha-hydroxyestrone ratio in premenopausal and postmenopausal women.
Experimental and clinical evidence suggests that 16alpha-hydroxylated estrogen metabolites, biologically strong estrogens, are associated with breast cancer risk, while 2-hydroxylated metabolites, with lower estrogenic activity, are weakly related to this disease. This study analyzes the association of breast cancer risk with estrogen metabolism, expressed as the ratio of 2-hydroxyestrone to 16alpha-hydroxyestrone, in a prospective nested case-control study. Between 1987 and 1992, 10,786 women (ages 35-69 years) were recruited to a prospective study on breast cancer in Italy, the "Hormones and Diet in the Etiology of Breast Cancer" (ORDET) study. Women with a history of cancer and women on hormone therapy were excluded at baseline. At recruitment, overnight urine was collected from all participants and stored at -80 degrees C. After an average of 5.5 years of follow-up, 144 breast cancer cases and four matched controls for each case were identified among the participants of the cohort. Among premenopausal women, a higher ratio of 2-hydroxyestrone to 16alpha-hydroxyestrone at baseline was associated with a reduced risk of breast cancer: women in the highest quintile of the ratio had an adjusted odds ratio (OR) for breast cancer of 0.58 [95% confidence interval (CI) = 0.25-1.34]. The corresponding adjusted OR in postmenopausal women was 1.29 (95% CI = 0.53-3.10). Results of this prospective study support the hypothesis that the estrogen metabolism pathway favoring 2-hydroxylation over 16alpha-hydroxylation is associated with a reduced risk of invasive breast cancer risk in premenopausal women.


Neuronal Computation Underlying Inferential Reasoning in Humans and Mice.
Every day we make decisions critical for adaptation and survival. We repeat actions with known consequences. But we also draw on loosely related events to infer and imagine the outcome of entirely novel choices. These inferential decisions are thought to engage a number of brain regions; however, the underlying neuronal computation remains unknown. Here, we use a multi-day cross-species approach in humans and mice to report the functional anatomy and neuronal computation underlying inferential decisions. We show that during successful inference, the mammalian brain uses a hippocampal prospective code to forecast temporally structured learned associations. Moreover, during resting behavior, coactivation of hippocampal cells in sharp-wave/ripples represent inferred relationships that include reward, thereby "joining-the-dots" between events that have not been observed together but lead to profitable outcomes. Computing mnemonic links in this manner may provide an important mechanism to build a cognitive map that stretches beyond direct experience, thus supporting flexible behavior.


Glypican-3-Specific CAR T Cells Coexpressing IL15 and IL21 Have Superior Expansion and Antitumor Activity against Hepatocellular Carcinoma.
Hepatocellular carcinoma (HCC) is the fourth most common cause of cancer-related death in the world, and curative systemic therapies are lacking. Chimeric antigen receptor (CAR)-expressing T cells induce robust antitumor responses in patients with hematologic malignancies but have limited efficacy in patients with solid tumors, including HCC. IL15 and IL21 promote T-cell expansion, survival, and function and can improve the antitumor properties of T cells. We explored whether transgenic expression of IL15 and/or IL21 enhanced glypican-3-CAR (GPC3-CAR) T cells' antitumor properties against HCC. We previously optimized the costimulation in GPC3-CARs and selected a second-generation GPC3-CAR incorporating a 4-1BB costimulatory endodomain (GBBz) for development. Here, we generated constructs encoding IL15, IL21, or both with GBBz (15.GBBz, 21.GBBz, and 21.15.GBBz, respectively) and examined the ability of transduced T cells to kill, produce effector cytokines, and expand in an antigen-dependent manner. We performed gene-expression and phenotypic analyses of GPC3-CAR T cells and CRISPR-Cas9 knockout of the TCF7 gene. Finally, we measured GPC3-CAR T-cell antitumor activity in murine xenograft models of GPC3+ tumors. The increased proliferation of 21.15.GBBz T cells was at least in part dependent on the upregulation and maintenance of TCF-1 (encoded by TCF7) and associated with a higher percentage of stem cell memory and central memory populations after manufacturing. T cells expressing 21.15.GBBz had superior in vitro and in vivo expansion and persistence, and the most robust antitumor activity in vivo These results provided preclinical evidence to support the clinical evaluation of 21.15.GPC3-CAR T cells in patients with HCC.


Glucose metabolism in obesity and type 2 diabetes.
The aim of this review is to provide a comprehensive summary of the subject of glucose metabolism in normal and obese subjects, and in those with type 2 diabetes. The following topics are discussed: Glucose and insulin metabolism, including characterization of the role of various organs in maintaining glucose homeostasis in the basal state and after food. The action of insulin and the concept of insulin resistance, its main characteristics as revealed by studies of glucose metabolism in liver and skeletal muscle, the two organs primarily involved. Changes in insulin secretion in obese subjects and in diabetics are reviewed. Finally, the development of diabetes as a consequence of increasing insulin resistance in the obese diabetic subject is discussed. The principal features--insulin resistance, elevated free fatty acid levels and glucotoxicity--are emphasized. A possible explanation of how obesity leads to diabetes, emphasizing the role played by upper body fat distribution, is provided.


3D extrusion printing of high drug loading immediate release paracetamol tablets.
The manufacture of immediate release high drug loading paracetamol oral tablets was achieved using an extrusion based 3D printer from a premixed water based paste formulation. The 3D printed tablets demonstrate that a very high drug (paracetamol) loading formulation (80% w/w) can be printed as an acceptable tablet using a method suitable for personalisation and distributed manufacture. Paracetamol is an example of a drug whose physical form can present challenges to traditional powder compression tableting. Printing avoids these issues and facilitates the relatively high drug loading. The 3D printed tablets were evaluated for physical and mechanical properties including weight variation, friability, breaking force, disintegration time, and dimensions and were within acceptable range as defined by the international standards stated in the United States Pharmacopoeia (USP). X-ray Powder Diffraction (XRPD) was used to identify the physical form of the active. Additionally, XRPD, Attenuated Total Reflectance Fourier Transform Infrared spectroscopy (ATR-FTIR) and differential scanning calorimetry (DSC) were used to assess possible drug-excipient interactions. The 3D printed tablets were evaluated for drug release using a USP dissolution testing type I apparatus. The tablets showed a profile characteristic of the immediate release profile as intended based upon the active/excipient ratio used with disintegration in less than 60 s and release of most of the drug within 5 min. The results demonstrate the capability of 3D extrusion based printing to produce acceptable high-drug loading tablets from approved materials that comply with current USP standards.


Long-term observation of morphologic and functional features of cat corneal endothelium after wounding.
(1) To test the hypothesis that corneas with enlarged endothelial cells (and thus less intercellular space) have decreased endothelial permeability to small polar solutes. (2) To measure corneal endothelial ouabain binding (Na+/K+ ATPase "pump site" density) and Descemet's membrane production after endothelial wounding. Bilateral specular microscopy and anterior segment fluorophotometry were performed at 2-month intervals for 1 year in ten cats after mechanically damaging the corneal endothelium in one eye of each. The measurements were repeated at 2 years in four cats and at 3 years in two cats. Eighteen months after wounding, endothelial ouabain binding was measured in both eyes of six cats. Transmission electron micrographs of Descemet's membrane were analyzed in both eyes of six cats at 18 months, two cats at 2 years, and two cats at 3 years after wounding. From 6 to 12 months after wounding, the endothelial permeability to carboxyfluorescein was significantly decreased (P < 0.05), and the mean endothelial cell size was significantly increased (P < 0.001) in the damaged eyes. The enlarged endothelial cells persisted in the few cats observed 2 and 3 years after wounding. There was no significant difference in endothelial ouabain binding between the damaged and control corneas in six cats tested 18 months after wounding. On subsequent histologic examination, a layer of abnormal Descemet's membrane was present in all ten wounded eyes, with additional normal Descemet's membrane posterior to it, between the abnormal layer and the endothelial cells. The results are consistent with the hypothesis that corneal endothelial permeability to small polar solutes varies directly with the amount of intercellular space available for diffusion across the monolayer. The results also confirm clinical reports of decreased endothelial permeability in corneas with enlarged endothelial cells. In histopathologic specimens, a layer of abnormal Descemet's membrane can be a historical marker for a period of endothelial damage and corneal decompensation.


A new potassium-based intermediate and its role in the desorption properties of the K-Mg-N-H system.
New insights into the reaction pathways of different potassium/magnesium amide-hydride based systems are discussed. In situ SR-PXD experiments were for the first time performed in order to reveal the evolution of the phases connected with the hydrogen releasing processes. Evidence of a new K-N-H intermediate is shown and discussed with particular focus on structural modification. Based on these results, a new reaction mechanism of amide-hydride anionic exchange is proposed.


Identification of sites responsible for the potentiating effect of niflumic acid on ClC-Ka kidney chloride channels.
ClC-K kidney Cl(-) channels are important for renal and inner ear transepithelial Cl(-) transport, and are potentially interesting pharmacological targets. They are modulated by niflumic acid (NFA), a non-steroidal anti-inflammatory drug, in a biphasic way: NFA activates ClC-Ka at low concentrations, but blocks the channel above approximately 1 mM. We attempted to identify the amino acids involved in the activation of ClC-Ka by NFA. We used site-directed mutagenesis and two-electrode voltage clamp analysis of wild-type and mutant channels expressed in Xenopus oocytes. Guided by the crystal structure of a bacterial CLC homolog, we screened 97 ClC-Ka mutations for alterations of NFA effects. Mutations of five residues significantly reduced the potentiating effect of NFA. Two of these (G167A and F213A) drastically altered general gating properties and are unlikely to be involved in NFA binding. The three remaining mutants (L155A, G345S and A349E) severely impaired or abolished NFA potentiation. The three key residues identified (L155, G345, A349) are localized in two different protein regions that, based on the crystal structure of bacterial CLC homologs, are expected to be exposed to the extracellular side of the channel, relatively close to each other, and are thus good candidates for being part of the potentiating NFA binding site. Alternatively, the protein region identified mediates conformational changes following NFA binding. Our results are an important step towards the development of ClC-Ka activators for treating Bartter syndrome types III and IV with residual channel activity.


Insulin-degrading enzyme prevents α-synuclein fibril formation in a nonproteolytical manner.
The insulin-degrading enzyme (IDE) degrades amyloidogenic proteins such as Amyloid β (Αβ) and Islet Amyloid Polypeptide (IAPP), i.e. peptides associated with Alzheimer's disease and type 2 diabetes, respectively. In addition to the protease activity normally associated with IDE function an additional activity involving the formation of stable, irreversible complexes with both Αβ and α-synuclein, an amyloidogenic protein involved in Parkinson's disease, was recently proposed. Here, we have investigated the functional consequences of IDE-α-synuclein interactions in vitro. We demonstrate that IDE in a nonproteolytic manner and at sub-stoichiometric ratios efficiently inhibits α-synuclein fibril formation by binding to α-synuclein oligomers making them inert to amyloid formation. Moreover, we show that, within a defined range of α-synuclein concentrations, interaction with α-synuclein oligomers increases IDE's proteolytic activity on a fluorogenic substrate. We propose that the outcomes of IDE-α-synuclein interactions, i.e. protection against α-synuclein amyloid formation and stimulated IDE protease activity, may be protective in vivo.


Ribosomal RNA (rRNA) sequences from 33 globally distributed mosquito species for improved metagenomics and species identification.
Total RNA sequencing (RNA-seq) is an important tool in the study of mosquitoes and the RNA viruses they vector as it allows assessment of both host and viral RNA in specimens. However, there are two main constraints. First, as with many other species, abundant mosquito ribosomal RNA (rRNA) serves as the predominant template from which sequences are generated, meaning that the desired host and viral templates are sequenced far less. Second, mosquito specimens captured in the field must be correctly identified, in some cases to the sub-species level. Here, we generate mosquito rRNA datasets which will substantially mitigate both of these problems. We describe a strategy to assemble novel rRNA sequences from mosquito specimens and produce an unprecedented dataset of 234 full-length 28S and 18S rRNA sequences of 33 medically important species from countries with known histories of mosquito-borne virus circulation (Cambodia, the Central African Republic, Madagascar, and French Guiana). These sequences will allow both physical and computational removal of rRNA from specimens during RNA-seq protocols. We also assess the utility of rRNA sequences for molecular taxonomy and compare phylogenies constructed using rRNA sequences versus those created using the gold standard for molecular species identification of specimens-the mitochondrial cytochrome c oxidase I (COI) gene. We find that rRNA- and COI-derived phylogenetic trees are incongruent and that 28S and concatenated 28S+18S rRNA phylogenies reflect evolutionary relationships that are more aligned with contemporary mosquito systematics. This significant expansion to the current rRNA reference library for mosquitoes will improve mosquito RNA-seq metagenomics by permitting the optimization of species-specific rRNA depletion protocols for a broader range of species and streamlining species identification by rRNA sequence and phylogenetics.


Substance Abuse and Addiction: Implications for Pain Management in Patients With Cancer .
Substance use disorders (SUDs) are chronic diseases that may complicate the nursing care of patients with cancer, affecting their ability to adhere to treatment protocols, responses to cancer pain, and use of opioids for analgesia. This article explores how the presence of an SUD may affect oncology nursing care and pain management. The PubMed and CINAHL® databases were searched for articles from 1980-2016 using the keywords cancer, cancer pain, addiction, substance abuse, and alcoholism. . SUD is a common comorbidity in patients with cancer that may play a role in disease etiology. Practice guidelines are suggested for the assessment and management of addiction to improve overall outcomes for patients. Effective treatments for SUDs and cancer pain exist, and recovery can lead to improvements in multiple aspects of patients' lives.


Previous sickness absence and current low perceived social support at work among employees in the general population: a historical cohort study.
Although sickness absence often is a process over time, most studies have treated the phenomenon as a discrete event and focused more on its causes than its consequences. We aimed to examine whether various patterns of previous long-term sickness absence were associated with current low perceived social support at work. This is a historical cohort study based on data from a population-based survey among Swedish employees (n=2581). The survey data were linked to official registries yielding data on sickness absence 1-7 years prior to the survey. The main finding was that previous sickness absence was associated with current low perceived social support at work. The highest odds for low social support were found among those who had a stable high level of sickness absence. The two indicators of perceived social support employed were somewhat differently associated with previous sickness absence: Recency of absence showed to be of importance for general support at the workplace and the relationship with colleagues and superiors. Experiencing that one's immediate superior rarely or never regards one's view was, on the other hand, mainly related to having had a high level of sickness absence, irrespective of recency. Our results indicate that recency and extent of previous sickness absence are related to perceived social support at work. Future research on the relationship between social support and sickness absence should use repeated measurements and acknowledge the possible bidirectional relationship.


Associations Between Cancer Screening Behavior and Complementary Medicine Use: Results of a National Cross-Sectional Survey of 9151 Australian Women.
Complementary medicine (CM) use has been found to influence the uptake of conventional cancer treatment. This study examines associations between CM use and cancer screening rates. Women aged 62 to 67 years from the Australian Longitudinal Study on Women's Health were surveyed regarding their use of cancer screening initiatives. Associations between cancer screening behavior and visits to CM practitioners were analyzed. Of the 9151 women, 9049 (98.9%) completed questions about cancer screening. A total of 65.1% of women had received a clinical skin examination, 54.3% colorectal cancer screening, 56.2% Pap test (within past 2 years), 83.3% mammogram (within past 2 years), 55.8% clinical breast examination, and 55.8% had conducted breast self-examination. Women who had consulted a massage therapist were more likely to undergo clinical skin examination ( P = .002), clinical breast examination ( P = .018), and mammogram ( P = .001). Women who had consulted a chiropractor were more likely to undergo a clinical skin examination ( P = .001), colorectal cancer screening ( P = .020), and mammogram ( P = .011). Women who had consulted an acupuncturist were more likely to undergo colorectal cancer screening ( P = .019), and those who consulted with an osteopath were more liable to have a Pap test ( P = .049). Women who visit CM practitioners are more likely to participate in cancer screening initiatives. Research is required to understand the current and potential role that CM practitioners (can) have as public health advocates, recommending preventative health measures such as cancer screening. Such an examination will help ensure optimal screening utilization and effective, timely care for all cancer patients.


A real-world study of effectiveness of intravitreal bevacizumab and ranibizumab injection for treating retinal diseases in Thailand.
To evaluate the effectiveness of intravitreal bevacizumab (IVB) and intravitreal ranibizumab (IVR) in actual practice for treating patients with retinal diseases in Thailand. A prospective, multi-centre, observational study was conducted among eight hospitals in their ophthalmology outpatient departments. Participants consisted of patients who had previously not received any IVB or IVR treatment between 2013 and 2014. The primary outcome measurement was the change in best-corrected visual acuity (BCVA) at the end of the follow-up period compared to baseline. There were 1629 treatment-naïve patients for the pro re nata (PRN) treatment pattern and 226 treatment-naive patients for the three-injections (3Inj) treatment pattern. BCVA improvements were found in 35% of the PRN group and 47% of the 3Inj group; however, it was not clinically meaningful between the IVB and IVR groups (P-value = 0.568 for PRN, P-value = 0.103 for 3Inj). A multivariable logistic regression (using the propensity score) showed that positive factors associated with vision improvement for the PRN pattern were the number of drug injections, having retinal vein occlusion, and under 60 years of age, while good BCVA at baseline was a negative predictive factor. For the 3Inj pattern, under 60 years of age and baseline BCVA were statistically significant predictors. Nonetheless, diabetes mellitus (DM) without other comorbidities was a statistically significant predictor of low response to vision improvement compared to DM with other comorbidities. This study was the first observational, prospective study to evaluate the real-life effectiveness of IVB and IVR in Thailand. The majority of participants who used IVB or IVR showed improvements in BCVA after treatment. Further evaluation such as long-term follow-ups and subsequent comparison of effectiveness between IVB and IVR should be investigated due to the limited sample of IVR patients. Thai Clinical Trial Registry TCTR20141002001 . Registered 02 October 2014 (retrospectively registered).


First Report of Ramularia didyma Causing a Leaf Spot on Ranunculus (Ranunculus asiaticus) Hybrids in California.
Ranunculus or Persian buttercups (Ranunculus asiaticus) are colorful, cool-season perennials or annuals grown as landscape bedding plants and for field-grown bulb and cut flower production in mild winter climates. In March of 2008, tan-to-brown lesions were observed on the foliage of containerized ranunculus growing in a greenhouse at a production and retail nursery in coastal San Mateo County, CA. Approximately 15% of the approximately 150 cv. Bloomingdale mixed shade ranunculus plants had leaf spot symptoms. Symptomatic plants were generally clustered together on two benches, with a double-flowered purple cultivar with picotee markings having the highest disease incidence and severity. Angular to round, necrotic lesions ranged between 0.25 and 1.0 cm in diameter and were often surrounded by chlorotic tissue. Whitish sporulation was observed on abaxial and adaxial sides of the larger lesions. Conidiophores were hyaline, aseptate, measured 45 to 75 × 3 to 5 μm (56 × 3.7 μm average), and were produced in fascicles on the leaf surface. One-celled fusiform to cylindrical conidia measured 22.5 to 30.0 × 9.0 to 15 μm (25.4 × 11.9 μm average) and two-celled conidia measured 26.0 to 47.5 × 10 to 14 μm (35.3 × 12.1 μm average). Most conidia were hyaline, although a few were pale brown. Morphologically, the fungus matched the description of Ramularia didyma Unger (1). Small (3-mm2) pieces were taken from the margin of the lesion, disinfested with 0.6% sodium hypochlorite for 2 min, and placed at room temperature on carrot piece agar. Colonies were white and grew slowly to approximately 3.5 cm in 25 days. No conidia were produced in culture. To conduct pathogenicity tests, the foliage of nine mixed-color ranunculus plants grown from tubers for 7 weeks at 16°C in a growth chamber were sprayed with an aqueous suspension of mycelia. Inoculum was produced by grinding five, 3.5-cm colonies on carrot piece agar in 120 ml of water with a mortar and pestle. The foliage of an equal number of plants was sprayed with water. Plants were incubated in a dew chamber at 20°C for 48 h in the dark and then moved into a growth chamber with a 12-h photoperiod where relative humidity was maintained at ~95% by placing plants over a tray of water and covering each plant group with a plastic tent. Small, angular spots developed on approximately half of the inoculated plants 20 days later when the fungus was reisolated from the lesions. No symptoms were observed on the noninoculated control plants. Sporulation was observed on diseased lesions following misting of plants and incubation in sealed plastic bags for an additional week. Pathogenicity tests were repeated with mycelia with similar results. Sequence of the internal transcribed spacer regions of the rDNA was deposited into GenBank (Accession No. HQ442297). The sequence matched R. coleosporii and R. carthami with 96% identity. R. didyma has been reported to cause a leaf spot on Ranunculus spp. in the United States (Delaware, Iowa, Indiana, Maryland, New York, and Vermont). To our knowledge, this is the first confirmed report of R. didyma on Ranunculus asiaticus in California. Introduction of this pathogen into commercial production fields could cause significant economic loss. The closest large-scale production fields are located approximately 430 miles south of San Mateo County near Carlsbad, CA where more than 50 acres of ranunculus are grown for cut flower and tuber production. Reference: (1) J. B. Ellis and B. M. Everhart. J. Mycol. 1:79, 1885.


Cysteinyl leukotrienes in chronic hyperplastic rhinosinusitis.
Our study was designed to demonstrate that, similar to asthma, eosinophilic chronic rhinosinusitis is a disease characterized by activation and the expression of cysteinyl leukotrienes (LTs). Nasal polyp tissue was evaluated from 28 consecutive individuals undergoing elective polypectomy in whom neutrophils were not present on pathologic examination. Tissue was analyzed for pathosis with particular attention to the presence of eosinophils. Patients with moderate to high levels of mucosal eosinophils were classified as having eosinophilic rhinosinusitis (ECRS). Those with few or absent mucosal eosinophils were classified as having noneosinophilic rhinosinusitis (NECRS). Cysteinyl LTs were quantified by a sensitive competitive enzyme immunoassay, and the levels of cysteinyl LTs were compared in the groups. There were 20 patients with ECRS and 8 patients with NECRS. Cysteinyl LTs were identified in polyp tissue from 24 of 28 subjects and were >5 pg/gm tissue in 15 of 28. The average level of LTC(4) in patients with few mucosal eosinophils was 38.3 pg/g. The average amount in those with moderate to large amounts of mucosal eosinophils was 36.7 pg/g. There was no significant difference between the 2 groups. ECRS and NECRS can be considered diseases of excessive expression of cysteinyl LTs. LT expression occurs in patients who demonstrate few or no mucosal eosinophils, which indicates that cell lines other than eosinophils may be responsible for LT production in chronic rhinosinusitis.


Lower levels of proteinuria are associated with elevated mortality in incident dialysis patients.
Proteinuria is a potent predictor of adverse events in general, although a few large studies have reported a J-shaped association between proteinuria and mortality in individuals with glomerular filtration rate <30 ml/min/1.73m2. However, this association has not been specifically evaluated among incident dialysis patients. Among 1,380 Japanese patients who initiated dialysis, we quantified the association of pre-dialysis dipstick proteinuria (negative/trace, 1+, 2+, and ≥3+) with mortality using Cox models adjusting for potential confounders, such as age, gender, clinical history of hypertension, diabetes, and cardiovascular disease. Mean age of study participants was 67.4 (SD 13.0) years, and 67.6% were men. The most common dipstick proteinuria category was ≥3+ (55.4%), followed by 2+ (31.2%), 1+ (9.9%), and negative or trace (3.5%). Patients with lower proteinuria level were older than those with higher proteinuria. Lower proteinuria was significantly associated with a higher risk of all-cause mortality, even after accounting for potential confounders (p for trend <0.001). In those with negative/trace dipstick proteinuria compared to those with dipstick proteinuria ≥3+, the adjusted hazard ratio was 2.60 [95% CI: 1.62-4.17] in the fully adjusted model. Similar findings were observed when analyses were restricted to patients older than 70 years, and when cardiovascular mortality and non-cardiovascular mortality were analyzed separately. In incident dialysis patients, pre-dialysis proteinuria was inversely associated with mortality risk. Although future studies are needed to identify mechanisms, our findings suggest the need to carefully interpret proteinuria in patients with incident dialysis.


Temporary disruption of the plasma membrane is required for c-fos expression in response to mechanical stress.
Mechanically stressed cells display increased levels of fos message and protein. Although the intracellular signaling pathways responsible for FOS induction have been extensively characterized, we still do not understand the nature of the primary cell mechanotransduction event responsible for converting an externally acting mechanical stressor into an intracellular signal cascade. We now report that plasma membrane disruption (PMD) is quantitatively correlated on a cell-by-cell basis with fos protein levels expressed in mechanically injured monolayers. When the population of PMD-affected cells in injured monolayers was selectively prevented from responding to the injury, the fos response was completely ablated, demonstrating that PMD is a requisite event. This PMD-dependent expression of fos protein did not require cell exposure to cues inherent in release from cell-cell contact inhibition or presented by denuded substratum, because it also occurred in subconfluent monolayers. Fos expression also could not be explained by factors released through PMD, because cell injury conditioned medium failed to elicit fos expression. Translocation of the transcription factor NF-kappaB into the nucleus may also be regulated by PMD, based on a quantitative correlation similar to that found with fos. We propose that PMD, by allowing a flux of normally impermeant molecules across the plasma membrane, mediates a previously unrecognized form of cell mechanotransduction. PMD may thereby lead to cell growth or hypertrophy responses such as those that are present normally in mechanically stressed skeletal muscle and pathologically in the cardiovascular system.