Gender-specific effect of smoking on upper tract urothelial carcinoma outcomes.
To evaluate the gender-specific differential effects of smoking habits and cumulative smoking exposure on outcomes in patients with upper tract urothelial carcinoma (UTUC) treated with radical nephroureterectomy (RNU). A total of 864 consecutive patients, comprising 553 (64%) men and 311 (36%) women, from five international institutions underwent RNU without neoadjuvant chemotherapy. Smoking history included smoking status (current, former or never), quantity of cigarettes per day (CPD), smoking duration in years and years since smoking cessation. Cumulative smoking exposure was categorized as light short-term (≤19 CPD and ≤19.9 years), moderate (all combinations except light short-term and heavy long-term), and heavy long-term (≥20 CPD and ≥20 years). Uni- and multivariable competing risk regression models were used to assess the associations with outcomes. Overall, 244 (28.2%), 297 (34.4%) and 323 (37.4%) patients were never, former and current smokers, respectively. There were no differences in smoking status, quantity and duration between the genders. In female ever smokers, 30 (9.6%), 121 (38.9%) and 67 (21.5%) were light short-term, moderate and heavy long-term smokers, respectively. Compared with men, female current smokers were more likely to experience disease recurrence in univariable analysis (P = 0.013). In heavy long-term smokers, female gender was significantly associated with disease recurrence (hazard ratio [HR] 1.7; P = 0.03) and cancer-specific mortality (HR 2.0; P = 0.009) in multivariable analysis that adjusted for standard clinico-pathological features. In female patients only, smoking quantity, duration and cumulative exposure were associated with disease recurrence and cancer-specific mortality on multivariable analyses (P ≤ 0.025). The impact of smoking on UTUC outcomes after RNU is gender-specific. Females who are current and heavy long-term smokers experience worse outcomes than their male counterparts. Further research is needed to elucidate the molecular mechanisms underlying the gender-specific differential effect of smoking on UTUC outcomes.


Prevalence and burden of fecal incontinence: a population-based study in women.
The epidemiology of fecal incontinence (FI) is incompletely understood. We report the prevalence, clinical spectrum, health care-seeking behavior, and quality of life (QOL) in community women with FI. A questionnaire was mailed to an age-stratified random sample of 5300 Olmsted County, Minnesota, women identified by the Rochester Epidemiology Project. Symptom severity was assessed by a validated scale, and impact on QOL was evaluated for subjects who had any FI during the past year. The prevalence of FI was calculated with direct age adjustment to the 2000 US white female population. Altogether, 2800 of 5300 women (53%) responded to the survey. The overall age-adjusted prevalence of FI in the past year was 12.1 per 100 (95% confidence interval, 11.0-13.1). The prevalence increased with age from 7 (third decade) to 22 (sixth decade) per 100 and was steady thereafter. Symptoms were mild (45%), moderate (50%), or severe (5%), and symptom severity was related to the impact of FI on QOL and physician-consulting behavior. Moderate to severe impact on >/=1 domain of QOL was reported by 6% with mild, 35% with moderate, and 82% with severe symptoms, whereas 5% with mild, 10% with moderate, and 48% with severe FI had consulted a physician for FI in the past year. More than 1 of 10 adult women in the population have FI; almost 1 of 15 have moderate to severe FI. FI significantly impacts QOL and prompts health care utilization predominantly in women with moderate to severe symptoms.


Remodeling the cardiac transcriptional landscape with diet.
The perception that soy food products and dietary supplements will have beneficial effects on cardiovascular health has led to a massive consumer market. However, we have previously noted that diet profoundly affects disease progression in a genetic model of hypertrophic cardiomyopathy (HCM). In this model, a soy-based diet negatively impacts cardiac function in male mice. Given the frequent connection between functional changes and transcriptional changes, we investigated the effect of diet (soy- vs. milk-based) on cardiac gene expression and how it is affected by the additional factors of sex and disease. We found that gene expression in the heart is altered more by diet than by sex or an inherited disease. We also found that the healthy male heart may be sensitized to dietary perturbations of gene expression in that it displays a gene expression profile more similar to diseased male and female hearts than to healthy female hearts. These observations may in part account for documented divergence in HCM phenotypes between males and females and between diets.


A Challenge-Based Approach to Body Weight-Supported Treadmill Training Poststroke: Protocol for a Randomized Controlled Trial.
Body weight support treadmill training protocols in conjunction with other modalities are commonly used to improve poststroke balance and walking function. However, typical body weight support paradigms tend to use consistently stable balance conditions, often with handrail support and or manual assistance. In this paper, we describe our study protocol, which involved 2 unique body weight support treadmill training paradigms of similar training intensity that integrated dynamic balance challenges to help improve ambulatory function post stroke. The first paradigm emphasized walking without any handrails or manual assistance, that is, hands-free walking, and served as the control group, whereas the second paradigm incorporated practicing 9 essential challenging mobility skills, akin to environmental barriers encountered during community ambulation along with hands-free walking (ie hands-free + challenge walking). We recruited individuals with chronic poststroke hemiparesis and randomized them to either group. Participants trained for 6 weeks on a self-driven, robotic treadmill interface that provided body weight support and a safe gait-training environment. We assessed participants at pre-, mid- and post 6 weeks of intervention-training, with a 6-month follow-up. We hypothesized greater walking improvements in the hands-free + challenge walking group following training because of increased practice opportunity of essential mobility skills along with hands-free walking. We assessed 77 individuals with chronic hemiparesis, and enrolled and randomized 30 individuals poststroke for our study (hands-free group=19 and hands-free + challenge walking group=20) from June 2012 to January 2015. Data collection along with 6-month follow-up continued until January 2016. Our primary outcome measure is change in comfortable walking speed from pre to post intervention for each group. We will also assess feasibility, adherence, postintervention efficacy, and changes in various exploratory secondary outcome measures. Additionally, we will also assess participant responses to a study survey, conducted at the end of training week, to gauge each group's training experiences. Our treadmill training paradigms, and study protocol represent advances in standardized approaches to selecting body weight support levels without the necessity for using handrails or manual assistance, while progressively providing dynamic challenges for improving poststroke ambulatory function during rehabilitation. ClinicalTrials.gov NCT02787759; https://clinicaltrials.gov/ct2/show/NCT02787759 (Archived by Webcite at http://www.webcitation.org/6yJZCrIea).


Effect of Statin Therapy on Outcomes of Patients With Acute Ischemic Stroke and Atrial Fibrillation.
Background There is insufficient evidence on the effect of statins, particularly high-intensity statins, in patients with acute ischemic stroke and atrial fibrillation. We investigated the impact of statins on the outcomes in these patients, including those who might be vulnerable to statin therapy and those without clinical atherosclerotic cardiovascular diseases. Methods and Results A total of 2153 patients with acute ischemic stroke and atrial fibrillation were enrolled in the present nationwide, multicenter, cohort study. The primary composite end point was the occurrence of net adverse clinical and cerebral events (NACCE; death from any cause, stroke, acute coronary syndrome, or major bleeding) over a 3-year period based on statin intensity. NACCE rates were lower in patients receiving low- to moderate-intensity (adjusted hazard ratio 0.64; 95% CI: 0.52-0.78) and high-intensity statins (hazard ratio 0.51; 95% CI 0.40-0.66) than in those not receiving statin therapy. High-intensity statins were associated with a lower risk for NACCE than low- to moderate-intensity statins (hazard ratio 0.76; 95% CI 0.59-0.96). Subgroup analyses showed that the differences in hazard ratio for 3-year NACCE favored statin use across all subgroups, including older patients, those with low cholesterol levels, patients receiving anticoagulants, and patients without clinical atherosclerotic cardiovascular diseases. Magnified benefits of high-intensity statins compared with low- to moderate-intensity statins were observed in patients who underwent revascularization therapy and those under 75 years of age. Conclusions Statins, particularly high-intensity statins, could reduce the risk for NACCE in patients with acute ischemic stroke and atrial fibrillation; this needs to be further explored in randomized controlled trials.


Alpha6-containing nicotinic acetylcholine receptors dominate the nicotine control of dopamine neurotransmission in nucleus accumbens.
Modulation of striatal dopamine (DA) neurotransmission plays a fundamental role in the reinforcing and ultimately addictive effects of nicotine. Nicotine, by desensitizing beta2 subunit-containing (beta2*) nicotinic acetylcholine receptors (nAChRs) on striatal DA axons, significantly enhances how DA is released by reward-related burst activity compared to nonreward-related tonic activity. This action provides a synaptic mechanism for nicotine to facilitate the DA-dependent reinforcement. The subfamily of beta2*-nAChRs responsible for these potent synaptic effects could offer a molecular target for therapeutic strategies in nicotine addiction. We explored the role of alpha6beta2*-nAChRs in the nucleus accumbens (NAc) and caudate-putamen (CPu) by observing action potential-dependent DA release from synapses in real-time using fast-scan cyclic voltammetry at carbon-fiber microelectrodes in mouse striatal slices. The alpha6-specific antagonist alpha-conotoxin-MII suppressed DA release evoked by single and low-frequency action potentials and concurrently enhanced release by high-frequency bursts in a manner similar to the beta2*-selective antagonist dihydro-beta-erythroidine (DHbetaE) in NAc, but less so in CPu. The greater role for alpha6*-nAChRs in NAc was not due to any confounding regional difference in ACh tone since elevated ACh levels (after the acetylcholinesterase inhibitor ambenonium) had similar outcomes in NAc and CPu. Rather, there appear to be underlying differences in nAChR subtype function in NAc and CPu. In summary, we reveal that alpha6beta2*-nAChRs dominate the effects of nicotine on DA release in NAc, whereas in CPu their role is minor alongside other beta2*-nAChRs (eg alpha4*), These data offer new insights to suggest striatal alpha6*-nAChRs as a molecular target for a therapeutic strategy for nicotine addiction.


Specificity of resistance to dengue virus isolates is associated with genotypes of the mosquito antiviral gene Dicer-2.
In contrast to the prevailing view that invertebrate immunity relies on broad-spectrum recognition and effector mechanisms, intrinsic genetic compatibility between invertebrate hosts and their pathogens is often highly specific in nature. Solving this puzzle requires a better understanding of the molecular basis underlying observed patterns of invertebrate host-pathogen genetic specificity, broadly referred to as genotype-by-genotype interactions. Here, we identify an invertebrate immune gene in which natural polymorphism is associated with isolate-specific resistance to an RNA virus. Dicer-2 (dcr2) encodes a key protein upstream of the RNA interference (RNAi) pathway, a major antiviral component of innate immunity in invertebrates. We surveyed allelic polymorphism at the dcr2 locus in a wild-type outbred population and in three derived isofemale families of the mosquito Aedes aegypti that were experimentally exposed to several, genetically distinct isolates of dengue virus. We found that dcr2 genotype was associated with resistance to dengue virus in a virus isolate-specific manner. By contrast, no such association was found for genotypes at two control loci flanking dcr2, making it likely that dcr2 contains the yet-unidentified causal polymorphism(s). This result supports the idea that host-pathogen compatibility in this system depends, in part, on a genotype-by-genotype interaction between dcr2 and the viral genome, and points to the RNAi pathway as a potentially important determinant of intrinsic insect-virus genetic specificity.


Heterosubtypic cross-protection correlates with cross-reactive interferon-gamma-secreting lymphocytes in the ferret model of influenza.
An effective universal vaccine for influenza will likely need to induce virus-specific T-cells, which are the major mediator of heterosubtypic cross-protection between different subtypes of influenza A virus. In this study we characterise the cell-mediated immune response in ferrets during heterosubtypic protection induced by low-dose H1N1 virus infection against an H3N2 virus challenge, given 4 weeks later. Although the ferrets were not protected against the infection by H3N2 virus, the duration of virus shedding was shortened, and clinical disease was markedly reduced. No cross-reactive neutralizing antibodies were detected, but cross-reactive interferon-gamma-secreting T cells were detected in the circulation prior to H3N2 challenge. These T-cells peaked at 11 days post-H1N1 infection, and were strongly induced in blood and in lung following H3N2 infection. The rapid induction of interferon-gamma-secreting cells in ferrets previously infected with H1N1 virus, but not in naïve ferrets, suggests induction of memory T-cells. These results are in accord with the observations that pre-existing cross-reactive T-cells correlate with protection in humans and have implications for outbreak modelling and universal vaccine design.


Optically transparent cathode for Co(III/II) mediated dye-sensitized solar cells based on graphene oxide.
Thin semitransparent films were fabricated on F-doped SnO(2) (FTO) from single-layer graphene oxide (GO) either pure or in a composite with graphene nanoplatelets. Electrocatalytic activity of prepared films was tested for the Co(bpy)(3)(3+/2+) redox couple in acetonitrile electrolyte solution. Pristine GO showed almost no activity, resembling the properties of basal plane pyrolytic graphite. However, electrochemical performance of graphene oxide improved dramatically upon chemical reduction with hydrazine and/or heat treatment. All GO-containing films were firmly bonded to FTO, which contrasted with the poor adhesion of sole graphene nanoplatelets to this support. The activity loss during long-term aging was considerably improved, too. Enhanced stability of GO-containing films together with high electrocatalytic activity is beneficial for application in a new generation of dye-sensitized solar cells employing Co(bpy)(3)(3+/2+) as the redox shuttle.


Adding Fish Oil to Whey Protein, Leucine, and Carbohydrate Over a Six-Week Supplementation Period Attenuates Muscle Soreness Following Eccentric Exercise in Competitive Soccer Players.
Soccer players often experience eccentric exercise-induced muscle damage given the physical demands of soccer match-play. Since long chain n-3 polyunsaturated fatty acids (n-3PUFA) enhance muscle sensitivity to protein supplementation, dietary supplementation with a combination of fish oil-derived n-3PUFA, protein, and carbohydrate may promote exercise recovery. This study examined the influence of adding n-3PUFA to a whey protein, leucine, and carbohydrate containing beverage over a six-week supplementation period on physiological markers of recovery measured over three days following eccentric exercise. Competitive soccer players were assigned to one of three conditions (2 × 200 mL): a fish oil supplement beverage (FO; n = 10) that contained n-3PUFA (1100 mg DHA/EPA-approximately 550 mg DHA, 550 mg EPA), whey protein (15 g), leucine (1.8 g), and carbohydrate (20 g); a protein supplement beverage (PRO; n = 10) that contained whey protein (15 g), leucine (1.8 g), and carbohydrate (20 g); and a carbohydrate supplement beverage (CHO; n = 10) that contained carbohydrate (24 g). Eccentric exercise consisted of unilateral knee extension/flexion contractions on both legs separately. Maximal force production was impaired by 22% during the 72-hour recovery period following eccentric exercise (p < 0.05). Muscle soreness, expressed as area under the curve (AUC) during 72-hour recovery, was less in FO (1948 ± 1091 mm × 72 h) than PRO (4640 ± 2654 mm × 72 h, p < 0.05) and CHO (4495 ± 1853 mm × 72 h, p = 0.10). Blood concentrations of creatine kinase, expressed as AUC, were ~60% lower in FO compared to CHO (p < 0.05) and tended to be lower (~39%, p = 0.07) than PRO. No differences in muscle function, soccer performance, or blood c-reactive protein concentrations were observed between groups. In conclusion, the addition of n-3PUFA to a beverage containing whey protein, leucine, and carbohydrate ameliorates the increase in muscle soreness and blood concentrations of creatine kinase following eccentric exercise in competitive soccer players.


Direct Analysis of Oil Additives by High-Field Asymmetric Waveform Ion Mobility Spectrometry-Mass Spectrometry Combined with Electrospray Ionization and Desorption Electrospray Ionization.
The analysis of corrosion inhibitors in the presence and absence of an oil matrix is reported using electrospray ionization (ESI) and desorption electrospray ionization (DESI), hyphenated with miniaturized high-field asymmetric waveform ion mobility spectrometry (FAIMS) and mass spectrometry (MS). The target analytes were successfully ionized in solution by ESI and directly from steel surfaces using DESI ambient ionization at levels ≥0.0004% w/w (4 ppm) in oil. Differences in the mass spectral profiles observed for the additive/oil mixture are attributed to differences between the ESI and DESI ionization processes. The use of FAIMS improved selectivity for ESI generated analyte ions through reduction in the chemical noise resulting from the oil matrix. DESI enabled the direct, rapid, native state interrogation of oil samples on steel surfaces without sample pretreatment, and the hyphenation of DESI with the miniaturized FAIMS enhanced the relative analyte responses of the surface-active corrosion inhibitors.


From meta-omics to causality: experimental models for human microbiome research.
Large-scale 'meta-omic' projects are greatly advancing our knowledge of the human microbiome and its specific role in governing health and disease states. A myriad of ongoing studies aim at identifying links between microbial community disequilibria (dysbiosis) and human diseases. However, due to the inherent complexity and heterogeneity of the human microbiome, cross-sectional, case-control and longitudinal studies may not have enough statistical power to allow causation to be deduced from patterns of association between variables in high-resolution omic datasets. Therefore, to move beyond reliance on the empirical method, experiments are critical. For these, robust experimental models are required that allow the systematic manipulation of variables to test the multitude of hypotheses, which arise from high-throughput molecular studies. Particularly promising in this respect are microfluidics-based in vitro co-culture systems, which allow high-throughput first-pass experiments aimed at proving cause-and-effect relationships prior to testing of hypotheses in animal models. This review focuses on widely used in vivo, in vitro, ex vivo and in silico approaches to study host-microbial community interactions. Such systems, either used in isolation or in a combinatory experimental approach, will allow systematic investigations of the impact of microbes on the health and disease of the human host. All the currently available models present pros and cons, which are described and discussed. Moreover, suggestions are made on how to develop future experimental models that not only allow the study of host-microbiota interactions but are also amenable to high-throughput experimentation.


Iron status, blood lipids and endogenous antioxidants in response to dietary iron levels in male and female rats.
The effect of dietary iron levels on iron status, blood lipids and endogenous antioxidants was investigated in male and female rats. Diets low in iron (15 mg/kg Fe; LFe) or high in iron (400 mg/kg Fe; HFe) were given to groups of male (n = 6) and female (n = 6) weanling rats for six weeks. In a second experiment the same dietary iron levels were fed to groups (n = 12) of males and females for seven months, during which colon tumours were induced. Indices of iron status, blood lipid levels and antioxidant enzyme activities were measured in both experiments. In the first experiment, indices of iron status were significantly higher in HFe rats and in females compared with males. Cholesterol and triglycerides were significantly higher in HFe rats and cholesterol was significantly higher in males. Plasma albumin and bilirubin levels and plasma caeruloplasmin activity were significantly higher in female rats. The second experiment confirmed the higher indices of iron status in HFe rats and in female rats, and also showed that plasma cholesterol levels were significantly higher in HFe rats. There were no consistent, significant differences over both experiments in activities of the antioxidant enzymes measured. Results show that higher dietary iron levels are associated with higher cholesterol levels in male and female rats. However cholesterol was found to be higher in male rats while iron status was higher in female rats. This indicates that factors other than iron status are responsible for the differences in cholesterol in male and female rats.


Physician characteristics and prescription drug use during pregnancy: a population-based study.
We aimed to explore the relationship between physician characteristics and their prescribing behavior regarding category D and X drugs for pregnant women by using a population-based data set in Taiwan. The sampled population for the study included 14,430 women. These women received a total of 198,420 prescriptions during pregnancy. We performed multivariate logistic regression analysis by using generalized estimated equations to assess the odds ratio (OR) of the prescription for categories D and X drugs among doctors after adjusting for maternal age and chronic disease. Of the total 198,420 prescriptions, 4.2% were prescribed category D and X drugs. The covariate-adjusted odds for physicians aged between 40 and 49 years and 50 and 59 years for prescribing category D and X drugs to pregnant women were 1.22 (95% confidence interval [95% CI], 1.15-1.31) and 1.51 (95% CI, 1.40-1.64) times that of physicians aged between 30 and 39 years, respectively. Male physicians were less likely to prescribe category D and X drugs to pregnant women than female physicians (OR, 0.69; 95% CI, 0.63-0.75). In addition, physicians specializing in "other" specialties were more likely (OR, 1.46; 95% CI, 1.41-1.54) to prescribe category D and X drugs compared with those specializing in obstetrics/gynecology, whereas physicians practicing in central Taiwan were less likely (OR, 0.85; 95% CI, 0.80-0.89) than their counterparts in other regions of Taiwan to prescribe category D and X drugs. We conclude that physician characteristics, including sex, age, specialty, and practice location, were associated with the prescription of category D and X drugs for pregnant women.


DNA sequence and structure requirements for cleavage of V(D)J recombination signal sequences.
Purified RAG1 and RAG2 proteins can cleave DNA at V(D)J recombination signals. In dissecting the DNA sequence and structural requirements for cleavage, we find that the heptamer and nonamer motifs of the recombination signal sequence can independently direct both steps of the cleavage reaction. Proper helical spacing between these two elements greatly enhances the efficiency of cleavage, whereas improper spacing can lead to interference between the two elements. The signal sequences are surprisingly tolerant of structural variation and function efficiently when nicks, gaps, and mismatched bases are introduced or even when the signal sequence is completely single stranded. Sequence alterations that facilitate unpairing of the bases at the signal/coding border activate the cleavage reaction, suggesting that DNA distortion is critical for V(D)J recombination.


Parents' perspectives on talking to preteenage children about sex.
Although parent-child communication about sex can significantly affect children's sexual behavior, many parents do not talk to their children about sex. Qualitative research can elucidate parents' attitudes toward and experiences with communicating with their children about sex. In 2007, 16 focus groups were conducted with 131 mothers and fathers of children aged 10-12 in three cities in different regions of the United States. Separate groups were conducted for mothers and fathers, and for black, white and Hispanic parents. Content analysis was used to identify core themes and patterns. Parents believed it is important to talk to their children about sex and believed that doing so can be effective, but many had not done so. Primary barriers were parents' perception that their children are too young and not knowing how to talk to their children about the subject. Parents found it easiest to talk to their children about sex if they had a good parent-child relationship, took advantage of opportunities to talk and began having the discussions when their children were very young. Some differences were noted by parents' race, ethnicity, gender and location. Interventions aimed at encouraging parents to talk to their children about sex should enhance parents' understanding of the stages of children's sexual development and focus on the parents of young children. In addition, interventions should support parents in a range of strategies that complement discussions about sex.


The evolution of genetic topologies.
This manuscript explores the simultaneous evolution of population genetic parameters and topological features within a population graph through a series of Monte Carlo simulations. I show that node centrality and graph breadth are significantly correlated to population genetic parameters Phi(ST) and M; (rho=-0.95;rho=-0.98, respectively), which are commonly used in quantifying among population genetic structure and isolation by distance. Next, the topological consequences of migration patterns are examined by contrasting N-island and stepping stone models of gene movement. Finally, I show how variation in migration rate influences the rate of formation of specific topological features with particular emphasis to the phase transition that occurs when populations begin to become fixed due to restricted movement of genes among populations. I close by discussing the utility of this method for the analysis of intraspecific genetic variation.


Characterization of the equine 2'-5' oligoadenylate synthetase 1 (OAS1) and ribonuclease L (RNASEL) innate immunity genes.
The mammalian OAS/RNASEL pathway plays an important role in antiviral host defense. A premature stop-codon within the murine Oas1b gene results in the increased susceptibility of mice to a number of flaviviruses, including West Nile virus (WNV). Mutations in either the OAS1 or RNASEL genes may also modulate the outcome of WNV-induced disease or other viral infections in horses. Polymorphisms in the human OAS gene cluster have been previously utilized for case-control analysis of virus-induced disease in humans. No polymorphisms have yet been identified in either the equine OAS1 or RNASEL genes for use in similar case-control studies. Genomic sequence for equine OAS1 was obtained from a contig assembly generated from a shotgun subclone library of CHORI-241 BAC 100I10. Specific amplification of regions of the OAS1 gene from 13 horses of various breeds identified 33 single nucleotide polymorphisms (SNP) and two microsatellites. RNASEL cDNA sequences were determined for 8 mammals and utilized in a phylogenetic analysis. The chromosomal location of the RNASEL gene was assigned by FISH to ECA5p17-p16 using two selected CHORI-241 BAC clones. The horse genomic RNASEL sequence was assembled. Specific amplification of regions of the RNASEL gene from 13 horses identified 31 SNPs. In this report, two dinucleotide microsatellites and 64 single nucleotide polymorphisms within the equine OAS1 and RNASEL genes were identified. These polymorphisms are the first to be reported for these genes and will facilitate future case-control studies of horse susceptibility to infectious diseases.


Clinical relevance of cluster analysis in phenotyping allergic rhinitis in a real-life study.
Disease stratification, using phenotypic characterization performed either by hypothesis- or data-driven methods, was developed to improve clinical decisions. However, cluster analysis has not been used for allergic rhinitis. To define clusters in allergic rhinitis and to compare them with ARIA (Allergic Rhinitis and Its Impact on Asthma), a hypothesis-driven approach. A French observational prospective multicenter study (EVEIL: Echelle visuelle analogique dans la rhinite allergique) was carried out on 990 patients consulting general practitioners for allergic rhinitis and treated as per clinical practice. In this study, changes in symptom scores, visual analogue scales and quality of life were measured at baseline and after 14 days of treatment. A post hoc analysis was performed to identify clusters of patients with allergic rhinitis – using Ward's hierarchical method – and to define their clinical relevance at baseline and after 14 days of treatment. The cluster approach was compared to the ARIA approach. Patients were clustered into 4 phenotypes which partly followed the ARIA classes. These phenotypes differed in their disease severity including symptoms and quality of life. Physicians in real-life practice prescribed medication regardless of the phenotype and severity, with the exception of patients with ocular symptoms. Prescribed treatments were comparable in hypothesis- and data-driven analyses. The prevalence of uncontrolled patients during treatment was similar in the 4 clusters, but was significantly different according to the ARIA classes. Cluster analysis using demographic and clinical parameters only does not appear to add relevant information for disease stratification in allergic rhinitis.


Intracellular coexpression of epidermal growth factor receptor, Her-2/neu, and p21ras in human breast cancers: evidence for the existence of distinctive patterns of genetic evolution that are common to tumors from different patients.
Multiparameter flow cytometry studies were performed on cells from the primary tumors of 94 patients with breast cancer. Correlated cellular measurements of cell DNA content, Her-2/neu, epidermal growth factor receptor (EGFR), and p21ras levels were performed on each of 5,000 to 100,000 cells from each tumor. When criteria for positivity were matched with those in common use for immunohistochemical studies, 28 of 94 (30%) breast cancers were classified as positive for Her-2/neu overexpression. When similar criteria were applied to the EGFR measurements, 23 of 94 (24%) cases were classified as positive for EGFR overexpression. Similarly, 23 of 94 (24%) cases were classified as positive for p21ras overexpression. By conventional flow cytometric criteria for DNA ploidy, 24 cases were diploid, 28 were tetraploid, and 42 were aneuploid. When the measurements were treated as separate sets of data, the only statistically significant correlations noted were the high frequency of diploid tumors, which did not overexpress any of the three oncogenes studied (P < 0.05), and an association between Her-2/neu overexpression and aneuploidy (P < 0.03). When the data were treated as correlated intracellular measurements, 90 of the 94 tumors studied contained a population of cells in which the intracellular levels of Her-2/neu expression were directly correlated with the levels of EGFR expression in the same cells. The ratio of Her-2/neu molecules to EGFR molecules in the same cells exceeded 1 in the majority of tetraploid and aneuploid cases and was close to or less than 1 in the majority of diploid cases. In nearly all tumors, p21ras overexpression was observed only in cells that overexpressed Her-2/neu, EGFR, or both, and p21ras levels per cell were more closely correlated with levels of EGFR per cell in the same cells than with Her-2/neu levels per cell. The data are consistent with a model in which heterodimerization of Her-2/neu and EGFR in individual cells is achieved by one of several genetic evolutionary pathways, all of which commonly lead to p21ras overexpression. The two major genetic evolutionary pathways identified in this study are an aneuploid, Her-2/neu overexpression-driven pathway seen in 59 of 94 tumors, and a diploid, EGFR overexpression-driven pathway seen in 19 of 94 tumors. All tumors with Her-2/neu:EGFR ratios greater than 2 contained an infiltrating ductal carcinoma component, whereas all infiltrating pure lobular carcinomas had Her-2/ neu:EGFR ratios that were less than 2. All of the genetic evolutionary pathways identified in this study were represented among the 11 tumors from patients who experienced early tumor recurrences.


Quantifying Graft Detachment after Descemet's Membrane Endothelial Keratoplasty with Deep Convolutional Neural Networks.
We developed a method to automatically locate and quantify graft detachment after Descemet's membrane endothelial keratoplasty (DMEK) in anterior segment optical coherence tomography (AS-OCT) scans. A total of 1280 AS-OCT B-scans were annotated by a DMEK expert. Using the annotations, a deep learning pipeline was developed to localize scleral spur, center the AS-OCT B-scans and segment the detached graft sections. Detachment segmentation model performance was evaluated per B-scan by comparing (1) length of detachment and (2) horizontal projection of the detached sections with the expert annotations. Horizontal projections were used to construct graft detachment maps. All final evaluations were done on a test set that was set apart during training of the models. A second DMEK expert annotated the test set to determine interrater performance. Mean scleral spur localization error was 0.155 mm, whereas the interrater difference was 0.090 mm. The estimated graft detachment lengths were in 69% of the cases within a 10-pixel (∼150 µm) difference from the ground truth (77% for the second DMEK expert). Dice scores for the horizontal projections of all B-scans with detachments were 0.896 and 0.880 for our model and the second DMEK expert, respectively. Our deep learning model can be used to automatically and instantly localize graft detachment in AS-OCT B-scans. Horizontal detachment projections can be determined with the same accuracy as a human DMEK expert, allowing for the construction of accurate graft detachment maps. Automated localization and quantification of graft detachment can support DMEK research and standardize clinical decision-making.


Novel Insights into the Adipokinome of Obese and Obese/Diabetic Mouse Models.
The group of adipokines comprises hundreds of biological active proteins and peptides released from adipose tissue. Alterations of those complex protein signatures are suggested to play a crucial role in the pathophysiology of multifactorial, metabolic diseases. We hypothesized that also the pathophysiology of type-2-diabetes is linked to the dysregulation of the adipocyte secretome. To test this, we investigated mouse models with monogenic defects in leptin signaling which are susceptible to adipositas (C57BL/6 Cg-Lepob (obob)) or adipositas with diabetes (C57BL/KS Cg-Leprdb (dbdb)) according to their genetic background. At the age of 17 weeks, visceral fat was obtained and primary murine adipocytes were isolated to harvest secretomes. Quantitative proteome analyses (LC-ESI-MS/MS) identified more than 800 potential secreted proteins. The secretome patterns revealed significant differences connected to the pathophysiology of obese mice. Pathway analyses indicated that these differences focus on exosome modelling, but failed to provide more precise specifications. To investigate the relationship of secretome data to insulin sensitivity, we examined the content of diabetogenic lipids, i.e., diacylglycerols (DAGs), identified as key players in lipid-induced insulin resistance. In contrast to obob mice, fat tissue of dbdb mice showed elevated DAG content, especially of DAG species with saturated fatty acid C16:0 and C18:0, while unsaturated fatty acid C16:1 were only changed in obob. Furthermore, DAG signatures of the models specifically correlate to secreted regulated adipokines indicating specific pathways. In conclusion, our data further support the concept that the fat tissue is an endocrine organ that releases bioactive factors corresponding to adipose tissue health status.


An update on LAMA/LABA combinations for COPD.
The management of symptoms of chronic obstructive pulmonary disease (COPD) typically involves the use of inhaled long-acting bronchodilators. Previously, we have reviewed the long-acting muscarinic antagonists (LAMAs) ▼aclidinium,1 ▼glycopyrronium,2 tiotropium3 and ▼umeclidinium,4 and the long-acting beta2 agonists (LABAs) indacaterol,5 ▼olodaterol,6 and ▼vilanterol (in combination with fluticasone) in the management of COPD.7 Four fixed-dose combinations of a LAMA with a LABA are now available, all indicated as maintenance bronchodilator treatment to relieve symptoms in adult patients with COPD. Here, we provide a brief overview of the four combinations and consider the place of a LAMA/LABA in people with COPD and any factors that might help to choose between them.


Spontaneous regression of hepatocellular carcinoma: a systematic review.
To estimate the actual frequency of spontaneous regression of hepatocellular carcinoma. A systematic review of the literature published during 1978-2007 has been carried out to identify randomized clinical trials of hepatocellular carcinoma that included a control arm receiving either placebo or best supportive care, and in which patients were followed prospectively for tumor response using predefined criteria. Data extraction was conducted independently by two investigators. A meta-analysis to provide a global estimation of regressions in the control arms was performed using an empiric Bayesian random-effects model. We identified 16 cases of regression (including minor and partial responses) in 10 phase III clinical trials. The rate of spontaneous objective partial regression among patients with hepatocellular carcinoma was 0.406% [95% confidence interval: 0.067-1.043%]. Although very infrequent, spontaneous regression is not an extraordinary event among patients with hepatocellular carcinoma. Therefore, individual responses to any given therapy should be assessed with caution and this fact may be considered at the time of calculating sample size of pilot clinical trials of new agents.


Finding fault: Criticism as a care management strategy and its impact on outcomes for dementia caregivers.
Despite a large literature on the stress process, little attention has focused on how caregivers for persons living with dementia (PLWDs) provide care and how this may impact care outcomes. Criticism is a management strategy caregivers may use to respond to behavioral symptoms. We consider whether criticism is associated with caregivers' mental health and service utilization. Data are drawn from the Advancing Caregiver Training intervention study including 256 informal caregivers living with a PLWD. In multiple linear regressions controlling for caregivers' demographics and PLWDs' clinical factors, we consider criticism (criticism subscale of the Dementia Management Strategies Scale) as a predictor of caregiver burden, depressive symptoms, desire to institutionalize the PLWD, level of frustration with care, and the number of home-based, social, and health services utilized. On average, 15% of the sample sometimes reported using criticism as a management strategy to manage the challenges of care. Greater use of criticism was associated with significantly more caregiver burden (β = 0.26, P < 0.001) and frustration with caregiving (β = 0.66, P < 0.001), but not depressive symptoms or a desire to institutionalize the PLWD. Criticism was also associated with significantly greater utilization of home-based (β = 0.14, P < 0.05) and social services (β = 0.15, P < 0.05), but not health care services. Criticism appears to be used by more burdened and frustrated caregivers. The association of criticism with social and home-based services potentially reflects a need for greater support among this group of caregivers. Behavioral interventions that can help caregivers manage behavioral symptoms with positive, empirically validated strategies may be helpful.


Depressive Symptoms in Inner-City Children With Asthma.
BACKGROUND: Asthma is a sometimes severe respiratory illness with an increased prevalence, especially in low-income, minority, and inner-city populations, over the past 2 decades. Prior reports have suggested a link between depression and asthma deaths. However, no studies have examined the relationship between objective measures of asthma severity and clinician-rated depressive symptom severity. METHOD: In this pilot study, 46 children receiving treatment at an inner-city asthma clinic were assessed with the Children's Depression Rating Scale, Revised (CDRS-R). The current percentage of forced expiratory volume in 1 second (FEV(1)%) predicted and the inhaled steroid dose were recorded, as were oral steroid use, emergency room visits, and hospitalizations in the preceding year. RESULTS: Depressive symptoms were common in this sample, with 30% (N = 14) of the participants having CDRS-R scores consistent with likely, very likely, or almost certain major depressive disorder. When mean CDRS-R scores were compared between the sample divided by these asthma severity measures, only hospitalizations in the past year was associated with higher depressive symptom scores (p =.03). CONCLUSION: These findings suggest that in the patient sample studied, depressive symptoms appear to be common. However, depressive symptom severity is related only to hospitalization, not other measures of asthma severity. Larger studies are needed to confirm these findings and determine if other variables such as family history of depression or subjective assessment of asthma severity explain the high prevalence of depressive symptoms in these patients.


Pancreaticoduodenectomy for pancreatic adenocarcinoma: postoperative adjuvant chemoradiation improves survival. A prospective, single-institution experience.
This study was designed to evaluate prospectively survival after pancreaticoduodenectomy for pancreatic adenocarcinoma, comparing two different postoperative adjuvant chemoradiation protocol to those of no adjuvant therapy. Based on limited data from the Gastrointestinal Tumor Study Group, adjuvant chemoradiation therapy has been recommended after pancreaticoduodenectomy for adenocarcinoma of the head, neck, or uncinate process of the pancrease. However, many patients continue to receive no such therapy. From October 1991 through September 1995, all patients with resected, pathologically confirmed adenocarcinoma of the head, neck, or uncinate process of the pancreas were reviewed by a multidisciplinary group (surgery, radiation oncology, medical oncology, and pathology) and were offered three options for postoperative treatment after pancreaticoduodenectomy: 1) standard therapy: external beam radiation therapy to the pancreatic bed (4000-4500 cGy) given with two 3-day fluorouracil (5-FU) courses and followed by weekly bolus 5-FU (500 mg/m2 per day) for 4 months; 2) intensive therapy: external beam radiation therapy to the pancreatic bed (5040-5760 cGy) with prophylactic hepatic irradiation (2340-2700 cGy) given with and followed by infusional 5-FU (200 mg/m2 per day) plus leucovorin (5 mg/m2 per day) for 5 of 7 days for 4 months; or 3) no therapy: no postoperative radiation therapy or chemotherapy. Pancreaticoduodenectomy was performed in 174 patients, with 1 in-hospital death (0.6%). Ninety-nine patients elected standard therapy, 21 elected intensive therapy, and 53 patients declined therapy. The three groups were comparable with respect to race, gender, intraoperative blood loss, tumor differentiation, lymph node status, tumor diameter, and resection margin status. Univariate analyses indicated that tumor diameter < 3 cm, intraoperative blood loss < 700 mL, absence of intraoperative blood transfusions, and use of adjuvant chemoradiation therapy were associated with significantly longer survival (p < 0.05). By Cox proportional hazards survival analysis, the most powerful predictors of outcome were tumor diameter, intraoperative blood loss, status of resection margins, and use of postoperative adjuvant therapy. The use of postoperative adjuvant chemoradiation therapy was a predictor of improved survival (median survival, 19.5 months compared to 13.5 months without therapy; p = 0.003). The intensive therapy group had no survival advantage when compared to that of the standard therapy group (median survival, 17.5 months vs. 21 months, p = not significant). Adjuvant chemoradiation therapy significantly improves survival after pancreaticoduodenectomy for adenocarcinoma of the head, neck, or uncinate process of the pancreas. Based on these survival data, standard adjuvant chemoradiation therapy appears to be indicated for patients treated by pancreaticoduodenectomy for adenocarcinoma of the head, neck, or uncinate process of the pancreas. Intensive therapy conferred no survival advantage over standard therapy in this analysis.


Flexible band versus rigid ring annuloplasty for tricuspid regurgitation: a systematic review and meta-analysis.
Up to 20% of patients have pre-discharge residual moderate to severe tricuspid regurgitation (TR) after tricuspid repair. Reoperations for recurrent TR carry high mortality rates, which emphasizes the importance of identifying the optimal technique for the surgical management of TR. The present study is a systematic review and meta-analysis that aims to compare short and long term survival and freedom from TR of flexible band ring versus rigid ring for annuloplasty of TR. We conducted a systematic review and meta-analysis of comparative studies to evaluate these procedures. A systematic search of the literature was performed from six electronic databases. Pooled meta-analysis was conducted using odds ratio (OR) and weighted mean difference (WMD). The rates of in-hospital mortality were not different between the two groups, with cumulative rates of 6.9% for flexible band and 7.3% for rigid ring (OR: 0.92; 95% CI: 0.49-1.71). Rates of stroke were also similar with 1.7% of flexible band and 1.3% of rigid rings suffering a perioperative stroke (OR: 1.29; 95% CI: 0.74-2.23). Rigid ring had significantly better freedom from grade ≥2 TR at 5 years (OR: 0.44; 95% CI: 0.20-0.99) and overall (P=0.005). There was no significant difference in overall rates of reoperation (P=0.232) and survival (P=0.086) between flexible band and rigid ring. Both rigid ring and flexible band offer acceptable outcomes for the treatment of TR. Compared to flexible band, rates of TR are stable after rigid ring annuloplasty and long term freedom from TR are superior for rigid ring devices. Large prospective randomized trials are required in order to validate these findings and assess for improvements in patient survival.


Decomposing morphologically complex words in a nonlinear morphology.
Most Hebrew words are composed of 2 intertwined morphemes: a triconsonantal root and a phonological word pattern. Previous research with conjugated verb forms has shown consistent priming from the verbal patterns, suggesting that verbal forms are automatically parsed by native speakers into their morphemic constituents. The authors investigated the decomposition process, focusing on the structural properties of verbal forms that are perceived and extracted during word recognition. The manipulations consisted of using verbal forms derived from "weak" roots that have one consonant missing in some of the forms. The results demonstrated that if 1 consonant is missing, the parsing system collapses, and there is no evidence for morphological priming. In contrast, when a random consonant is inserted into the weak form, the verbal-pattern priming re-emerges. This outcome suggests that the constraint imposed on the decomposition process is primarily structural and abstract. Moreover, the all-or-none pattern of results is characteristic of rule-based behavior and not of simple correlational systems.


Complex assembly, crystallization and preliminary X-ray crystallographic analysis of the human Rod-Zwilch-ZW10 (RZZ) complex.
The spindle-assembly checkpoint (SAC) monitors kinetochore-microtubule attachment during mitosis. In metazoans, the three-subunit Rod-Zwilch-ZW10 (RZZ) complex is a crucial SAC component that interacts with additional SAC-activating and SAC-silencing components, including the Mad1-Mad2 complex and cytoplasmic dynein. The RZZ complex contains two copies of each subunit and has a predicted molecular mass of ∼800 kDa. Given the low abundance of the RZZ complex in natural sources, its recombinant reconstitution was attempted by co-expression of its subunits in insect cells. The RZZ complex was purified to homogeneity and subjected to systematic crystallization attempts. Initial crystals containing the entire RZZ complex were obtained using the sitting-drop method and were subjected to optimization to improve the diffraction resolution limit. The crystals belonged to space group P3₁ (No. 144) or P3₂ (No. 145), with unit-cell parameters a = b = 215.45, c = 458.7 Å, α = β = 90.0, γ = 120.0°.


Absence of S6K1 protects against age- and diet-induced obesity while enhancing insulin sensitivity.
Elucidating the signalling mechanisms by which obesity leads to impaired insulin action is critical in the development of therapeutic strategies for the treatment of diabetes. Recently, mice deficient for S6 Kinase 1 (S6K1), an effector of the mammalian target of rapamycin (mTOR) that acts to integrate nutrient and insulin signals, were shown to be hypoinsulinaemic, glucose intolerant and have reduced beta-cell mass. However, S6K1-deficient mice maintain normal glucose levels during fasting, suggesting hypersensitivity to insulin, raising the question of their metabolic fate as a function of age and diet. Here, we report that S6K1-deficient mice are protected against obesity owing to enhanced beta-oxidation. However on a high fat diet, levels of glucose and free fatty acids still rise in S6K1-deficient mice, resulting in insulin receptor desensitization. Nevertheless, S6K1-deficient mice remain sensitive to insulin owing to the apparent loss of a negative feedback loop from S6K1 to insulin receptor substrate 1 (IRS1), which blunts S307 and S636/S639 phosphorylation; sites involved in insulin resistance. Moreover, wild-type mice on a high fat diet as well as K/K A(y) and ob/ob (also known as Lep/Lep) mice-two genetic models of obesity-have markedly elevated S6K1 activity and, unlike S6K1-deficient mice, increased phosphorylation of IRS1 S307 and S636/S639. Thus under conditions of nutrient satiation S6K1 negatively regulates insulin signalling.


Impact of Transcendental Meditation on cardiovascular function at rest and during acute stress in adolescents with high normal blood pressure.
This study examined the impact of the Transcendental Meditation (TM) program on cardiovascular (CV) reactivity in adolescents with high normal BP. Thirty-five adolescents [34 African Americans (AAs), 1 Caucasian American (CA); ages 15-18 years] with resting systolic blood pressure (SBP) between the 85th and 95th percentile for their age and gender on three consecutive occasions, were randomly assigned to either TM (n=17) or health education control (CTL, n=18) groups. The TM group engaged in 15-min meditation twice each day for 2 months including sessions during school lunch break. Primary CV outcome measures were changes in blood pressure (BP), heart rate (HR), and cardiac output (CO) at rest and in response to two laboratory stressors, a simulated car driving stressor and an interpersonal social stressor interview. The TM group exhibited greater decreases in resting SBP (P<.03) from pre- to postintervention, compared to the CTL group. The TM group exhibited greater decreases from pre- to postintervention in SBP, HR, and CO reactivity (P's<.03) to the simulated car driving stressor, and in SBP reactivity (P<.03) to the social stressor interview. The TM program appears to have a beneficial impact upon CV functioning at rest and during acute laboratory stress in adolescents at-risk for hypertension.


Estimated Prevalence of Autism and Other Developmental Disabilities Following Questionnaire Changes in the 2014 National Health Interview Survey.
The developmental disabilities questions in the 2014 National Health Interview Survey (NHIS) were changed from previous years, including question reordering and a new approach to asking about autism spectrum disorder (ASD). This report examines survey-based estimates of the lifetime prevalence of ASD, intellectual disability (ID), and any other developmental delay (other DD) following the inclusion of a standalone ASD question, the inclusion of specific diagnoses in the ASD question, and the ASD question preceding the other DD question, and compares them with estimates from previous years. In NHIS, one child is randomly selected from each family to be the subject of detailed questions on health conditions, functional limitations, and health care utilization. Parents are asked if a doctor or health professional had ever told them that their child had each of a series of developmental disabilities. Prevalence estimates of ASD, ID, and other DD for children aged 3–17 years were calculated using data collected in 2011–2014. The estimated prevalence of ASD based on 2014 data was 2.24%, a significant increase from the estimated annualized prevalence of 1.25% based on 2011–2013 data. In contrast, the prevalence of other DD declined significantly from 4.84% based on 2011–2013 data to 3.57% based on 2014 data. The prevalence of ID did not significantly change from 2011–2013 (1.27%) to 2014 (1.10%). The prevalence of having any of the three conditions was constant across survey years. The revised question ordering and new approach to asking about developmental disabilities in the 2014 NHIS likely affected the prevalence estimates of these conditions. In previous years, it is likely that some parents of children diagnosed with ASD reported this developmental disability as other DD instead of, or in addition to, ASD. Following these changes, the 2014 ASD estimate was more similar to ASD prevalence estimates from other sources.


Mucositis-related morbidity and resource utilization in head and neck cancer patients receiving radiation therapy with or without chemotherapy.
The objective of this study was to estimate health care-resource utilization in head and neck cancer (HNC) patients. This was a prospective, longitudinal, multicenter, noninterventional study of mucositis in patients receiving radiation with or without chemotherapy for HNC. Mouth and throat soreness and functional impairment were measured using the Oral Mucositis Weekly Questionnaire-HNC. Resource utilization data were obtained from patient interviews and recorded from the patient's medical chart. Seventy-five patients were enrolled from six centers. Fifty (67%) patients received concurrent chemoradiation therapy; 34 (45%) received intensity-modulated radiation therapy. Over the course of treatment, 57 (76%) patients reported severe mouth and throat soreness. Pain and functional impairment because of mouth and throat soreness increased during the course of therapy despite the use of opioid analgesics in 64 (85%) of the patients. Complications of radiation therapy resulted in increased patient visits to physicians, nurses, and nutritionists. Thirty-eight (51%) patients had a feeding tube placed. Twenty-eight patients (37%) were hospitalized, five of whom were hospitalized twice; of the 33 admissions, 10 (30%) were designated as secondary to mucositis by their treating physician. Mean length of hospitalization was 4.9 days (range: 1-16). This study demonstrates that mucositis-related pain and functional impairment is associated with increased use of costly health resources. Effective treatments to reduce the pain and functional impairment of oral mucositis are needed in this patient population.


The relationship between weight status and emotional and behavioral problems in Spanish preschool children.
To examine cross-sectional and longitudinal associations between behavioral problems and weight status, considering body mass index (BMI) z-scores and overweight status, in a community sample of preschoolers. The Strengths and Difficulties Questionnaire and the Diagnostic Interview for Children and Adolescents were administered to 611 parents. Adjusted general linear models and binary logistic regressions were used. Children who were overweight and had a higher BMI were at increased risk of peer problems and attention-deficit/hyperactivity disorder (ADHD) symptoms. Prospective analyses showed that a higher BMI at the age of 3 years was predictive of peer problems at ages 4 and 5 years and hyperactivity and ADHD symptoms at the age of 4 years. This is the first study using a diagnostic-based instrument that shows a relationship between weight status and ADHD symptoms in preschoolers. Overweight children might benefit from screening for behavioral disorders and peer relationship problems.


A VLP-based vaccine targeting domain III of the West Nile virus E protein protects from lethal infection in mice.
Since its first appearance in the USA in 1999, West Nile virus (WNV) has spread in the Western hemisphere and continues to represent an important public health concern. In the absence of effective treatment, there is a medical need for the development of a safe and efficient vaccine. Live attenuated WNV vaccines have shown promise in preclinical and clinical studies but might carry inherent risks due to the possibility of reversion to more virulent forms. Subunit vaccines based on the large envelope (E) glycoprotein of WNV have therefore been explored as an alternative approach. Although these vaccines were shown to protect from disease in animal models, multiple injections and/or strong adjuvants were required to reach efficacy, underscoring the need for more immunogenic, yet safe DIII-based vaccines. We produced a conjugate vaccine against WNV consisting of recombinantly expressed domain III (DIII) of the E glycoprotein chemically cross-linked to virus-like particles derived from the recently discovered bacteriophage AP205. In contrast to isolated DIII protein, which required three administrations to induce detectable antibody titers in mice, high titers of DIII-specific antibodies were induced after a single injection of the conjugate vaccine. These antibodies were able to neutralize the virus in vitro and provided partial protection from a challenge with a lethal dose of WNV. Three injections of the vaccine induced high titers of virus-neutralizing antibodies, and completely protected mice from WNV infection. The immunogenicity of DIII can be strongly enhanced by conjugation to virus-like particles of the bacteriophage AP205. The superior immunogenicity of the conjugate vaccine with respect to other DIII-based subunit vaccines, its anticipated favourable safety profile and low production costs highlight its potential as an efficacious and cost-effective prophylaxis against WNV.


Random-effects, fixed-effects and the within-between specification for clustered data in observational health studies: a simulation study.
When unaccounted-for group-level characteristics affect an outcome variable, traditional linear regression is inefficient and can be biased. The random- and fixed-effects estimators (RE and FE, respectively) are two competing methods that address these problems. While each estimator controls for otherwise unaccounted-for effects, the two estimators require different assumptions. Health researchers tend to favor RE estimation, while researchers from some other disciplines tend to favor FE estimation. In addition to RE and FE, an alternative method called within-between (WB) was suggested by Mundlak in 1978, although is utilized infrequently. We conduct a simulation study to compare RE, FE, and WB estimation across 16,200 scenarios. The scenarios vary in the number of groups, the size of the groups, within-group variation, goodness-of-fit of the model, and the degree to which the model is correctly specified. Estimator preference is determined by lowest mean squared error of the estimated marginal effect and root mean squared error of fitted values. Although there are scenarios when each estimator is most appropriate, the cases in which traditional RE estimation is preferred are less common. In finite samples, the WB approach outperforms both traditional estimators. The Hausman test guides the practitioner to the estimator with the smallest absolute error only 61% of the time, and in many sample sizes simply applying the WB approach produces smaller absolute errors than following the suggestion of the test. Specification and estimation should be carefully considered and ultimately guided by the objective of the analysis and characteristics of the data. The WB approach has been underutilized, particularly for inference on marginal effects in small samples. Blindly applying any estimator can lead to bias, inefficiency, and flawed inference.


Anti-CTLA-4 therapy broadens the melanoma-reactive CD8+ T cell response.
Anti-CTLA-4 treatment improves the survival of patients with advanced-stage melanoma. However, although the anti-CTLA-4 antibody ipilimumab is now an approved treatment for patients with metastatic disease, it remains unknown by which mechanism it boosts tumor-specific T cell activity. In particular, it is unclear whether treatment amplifies previously induced T cell responses or whether it induces new tumor-specific T cell reactivities. Using a combination ultraviolet (UV)-induced peptide exchange and peptide-major histocompatibility complex (pMHC) combinatorial coding, we monitored immune reactivity against a panel of 145 melanoma-associated epitopes in a cohort of patients receiving anti-CTLA-4 treatment. Comparison of pre- and posttreatment T cell reactivities in peripheral blood mononuclear cell samples of 40 melanoma patients demonstrated that anti-CTLA-4 treatment induces a significant increase in the number of detectable melanoma-specific CD8 T cell responses (P = 0.0009). In striking contrast, the magnitude of both virus-specific and melanoma-specific T cell responses that were already detected before start of therapy remained unaltered by treatment (P = 0.74). The observation that anti-CTLA-4 treatment induces a significant number of newly detected T cell responses-but only infrequently boosts preexisting immune responses-provides strong evidence for anti-CTLA-4 therapy-enhanced T cell priming as a component of the clinical mode of action.


Site-specific Patient-reported Outcome Measures for Hand Conditions: Systematic Review of Development and Psychometric Properties.
There are a number of site-specific patient-reported outcome measures (PROMs) for hand conditions used in clinical practice and research for assessing the efficacy of surgical and nonsurgical interventions. The most commonly used hand-relevant PROMs are as follows: Disabilities of the Arm, Shoulder and Hand (DASH), QuickDASH (qDASH), Michigan Hand Questionnaire (MHQ), Patient Evaluation Measure (PEM), Upper Extremity Functional Index (UEFI), and Duruoz Hand Index (DHI). There has been no systematic evaluation of the published psychometric properties of these PROMs. A PRISMA-compliant systematic review of the development and validation studies of these hand PROMs was prospectively registered in PROSPERO and conducted to assess their psychometric properties. A search strategy was applied to Medline, Embase, PsycINFO, and CINAHL. Abstract screening was performed in duplicate. Assessment of psychometric properties was performed. The search retrieved 943 articles, of which 54 articles met predefined inclusion criteria. There were 19 studies evaluating DASH, 8 studies evaluating qDASH, 13 studies evaluating MHQ, 5 studies evaluating UEFI, 4 studies evaluating PEM, and 5 studies evaluating DHI. Assessment of content validity, internal consistency, construct validity, reproducibility, responsiveness, floor/ceiling effect, and interpretability for each PROM is described. The psychometric properties of the most commonly used PROMs in hand research are not adequately described in the published literature. DASH, qDASH, and MHQ have the best-published psychometric properties, though they have either some poor psychometric performance or incompletely studied psychometric properties. There are more limited published data describing the psychometric properties of the UEFI, PEM, and DHI.


gamma-Acetylenic GABA in tardive dyskinesia.
Brain gamma-aminobutyric acid (GABA) has been proposed to play a role in the modulation of extrapyramidal motor function. The effects of increasing brain GABA with gamma-acetylenic GABA (GAG), a drug that inhibits GABA transaminase, were evaluated in ten patients with stable tardive dyskinesia during a blind placebo-controlled trial. Drug effects during active treatment and two placebo periods were evaluated by scoring randomly sequenced videotapes of tardive dyskinesia and parkinsonian symptoms recorded weekly during a standardized examination. Tardive dyskinesia was significantly reduced, and preexisting parkinsonism increased slightly. The largest decrease in tardive dyskinesia symptoms occurred in patients receiving higher neuroleptic doses, suggesting an interaction between GABA and dopamine. Prolactin values increased but growth hormone values were unchanged. Psychiatric symptoms were also unchanged during GAG treatment.


Genomic Spectrum and Phenotypic Heterogeneity of Human IL-21 Receptor Deficiency.
Biallelic inactivating mutations in IL21R causes a combined immunodeficiency that is often complicated by cryptosporidium infections. While eight IL-21R-deficient patients have been reported previously, the natural course, immune characteristics of disease, and response to hematopoietic stem cell transplantation (HSCT) remain to be comprehensively examined. In our study, we have collected clinical histories of 13 patients with IL-21R deficiency from eight families across seven centers worldwide, including five novel patients identified by exome or NGS panel sequencing. Eight unique mutations in IL21R were identified in these patients, including two novel mutations. Median age at disease onset was 2.5 years (0.5-7 years). The main clinical manifestations were recurrent bacterial (84.6%), fungal (46.2%), and viral (38.5%) infections; cryptosporidiosis-associated cholangitis (46.2%); and asthma (23.1%). Inflammatory skin diseases (15.3%) and recurrent anaphylaxis (7.9%) constitute novel phenotypes of this combined immunodeficiency. Most patients exhibited hypogammaglobulinemia and reduced proportions of memory B cells, circulating T follicular helper cells, MAIT cells and terminally differentiated NK cells. However, IgE levels were elevated in 50% of IL-21R-deficient patients. Overall survival following HSCT (6 patients, mean follow-up 1.8 year) was 33.3%, with pre-existing organ damage constituting a negative prognostic factor. Mortality of non-transplanted patients (n = 7) was 57.1%. Our detailed analysis of the largest cohort of IL-21R-deficient patients to date provides in-depth clinical, immunological and immunophenotypic features of these patients, thereby establishing critical non-redundant functions of IL-21/IL-21R signaling in lymphocyte differentiation, humoral immunity and host defense against infection, and mechanisms of disease pathogenesis due to IL-21R deficiency. Outcome following HSCT depends on prior chronic infections and organ damage, which should thus be considered as early as possible following molecular diagnosis.


Public health measures to control tuberculosis in low-income countries: ethics and human rights considerations.
In low-income countries, tuberculosis (TB) control measures should be guided by ethical concerns and human rights obligations. Control programs should consider the principles of necessity, reasonableness and effectiveness of means, proportionality, distributive justice, and transparency. Certain measures-detention, infection control, and treatment to prevent transmission-raise particular concerns. While isolation is appropriate under certain circumstances, quarantine is never an acceptable control measure for TB, and any detention must be limited by necessity and conducted humanely. States have a duty to implement hospital infection control to the extent of their available resources and to provide treatment to health care workers (HCWs) infected on the job. HCWs, in turn, have an obligation to provide care unless conditions are unreasonably and unforeseeably unsafe. Finally, states have an obligation to provide adequate access to treatment, as a means of preventing transmission, as broadly as possible and in a non-discriminatory fashion. Along with treatment, states should provide support to increase treatment adherence and retention with respect for patient privacy and autonomy. Compulsory treatment is almost never acceptable. Governments should take care to respect human rights and ethical obligations as they execute TB control programs.


Laser instrument for detecting water ripple slopes.
An optical instrument for use in the study of wind induced ripple on the water surface of a wind and wave tank is described. The instrument measures the tip angle of the projected surface normal vector in the down-wind and the cross-wind vertical planes independent of water height, thereby allowing studies of wind induced ripple on lower frequency, mechanical waves. Underwater hardware is reduced to one mirror. Each projected angle appears on a separate output channel as an analog voltage. An in situ calibration technique used prior to recording calibrates the over-all system. Sample output records are shown, and system performance is discussed.


A high-resolution reference genetic map positioning 8.8 K genes for the conifer white spruce: structural genomics implications and correspondence with physical distance.
Over the last decade, extensive genetic and genomic resources have been developed for the conifer white spruce (Picea glauca, Pinaceae), which has one of the largest plant genomes (20 Gbp). Draft genome sequences of white spruce and other conifers have recently been produced, but dense genetic maps are needed to comprehend genome macrostructure, delineate regions involved in quantitative traits, complement functional genomic investigations, and assist the assembly of fragmented genomic sequences. A greatly expanded P. glauca composite linkage map was generated from a set of 1976 full-sib progeny, with the positioning of 8793 expressed genes. Regions with significant low or high gene density were identified. Gene family members tended to be mapped on the same chromosomes, with tandemly arrayed genes significantly biased towards specific functional classes. The map was integrated with transcriptome data surveyed across eight tissues. In total, 69 clusters of co-expressed and co-localising genes were identified. A high level of synteny was found with pine genetic maps, which should facilitate the transfer of structural information in the Pinaceae. Although the current white spruce genome sequence remains highly fragmented, dozens of scaffolds encompassing more than one mapped gene were identified. From these, the relationship between genetic and physical distances was examined and the genome-wide recombination rate was found to be much smaller than most estimates reported for angiosperm genomes. This gene linkage map shall assist the large-scale assembly of the next-generation white spruce genome sequence and provide a reference resource for the conifer genomics community.


Socioeconomic constraints to effective management of Burkitt's lymphoma in south-eastern Nigeria.
This paper presents health outcomes and associated socioeconomic factors of 41 children admitted to a tertiary care institution in south-east Nigeria with Burkitt's lymphoma (BL) between 1987 and 2004. BL responds well to chemotherapy and does not pose a significant threat to health in industrialized nations. However, in resource-poor settings where it is endemic, socioeconomic factors significantly affect access to care for affected children, making this readily treatable condition a cause of considerable distress and early death in affected children. Half of the children reported in this paper presented with late stage disease. Although laboratory facilities were available, they were not accessible to all the children. Nearly a quarter of parents of these children could not afford the cost of confirmatory tests, and about a fifth (n = 8; 19.5%) of the children received no chemotherapy because of their parents' inability to pay. Only 21 of 41 children (51.2%) remained on treatment long enough (at least 12 weeks) to enable them to be confirmed either as short-term cure (n = 9; 64.3%), or as early relapse (n = 2; 4.9%). Owing to financial constraint, 13 of the parents (31.7%) withdrew their children against medical advice (n = 7; 17.1%) or left the hospital (n = 6; 14.6%). To address the challenge posed by these factors, we call for the establishment of a regional BL programme in Africa to help establish a critical mass of resources (human and material) to facilitate the development of an effective and accessible control programme in the region.


Search for W Boson Decays to Three Charged Pions.
For the first time, a search for the rare decay of the W boson to three charged pions has been performed. Proton-proton collision data recorded by the CMS experiment at a center-of-mass energy of 13 TeV, corresponding to an integrated luminosity of 77.3 fb^{-1}, have been analyzed. No significant excess is observed above the background expectation. An upper limit of 1.01×10^{-6} is set at 95% confidence level on the branching fraction of the W boson to three charged pions. This provides a strong motivation for theoretical calculations of this branching fraction.


Bruton's tyrosine kinase regulates apoptosis and JNK/SAPK kinase activity.
Mast cells derived from Bruton's tyrosine kinase (Btk)-defective xid or btk null mice showed greater expansion in culture containing interleukin-3 (IL-3) than those from wild-type (wt) mice. Although the proliferative response to IL-3 was not significantly different between the wt and xid mast cells, xid and btk null mast cells died by apoptosis more slowly than their wt counterparts upon IL-3 deprivation. Consistent with these findings, the apoptosis-linked c-Jun N-terminal kinase/stress-activated protein kinase (JNK) activity was compromised in these btk-mutated cells upon Fc(epsilon)RI crosslinking or upon stimulation with IL-3 or with stem cell factor. p38 activity was less severely, but significantly, affected by btk mutation, whereas extracellular signal-regulated kinases were not affected by the same mutation. Btk-mediated regulation of apoptosis and JNK activity was confirmed by reconstitution of btk null mutant mast cells with the wt btk cDNA. Furthermore, growth factor withdrawal induced the activation and sustained activity of JNK in wt mast cells, while JNK activity was consistently lower in btk-mutated mast cells. These results support the notion that Btk regulates apoptosis through the JNK activation.


Sanitation practices and perceptions in Kakuma refugee camp, Kenya: Comparing the status quo with a novel service-based approach.
Globally, an estimated 2.5 billion people lack access to improved sanitation. Unimproved sanitation increases the risk of morbidity and mortality, especially in protracted refugee situations where sanitation is based on pit latrine use. Once the pit is full, waste remains in the pit, necessitating the construction of a new latrine, straining available land and funding resources. A viable, sustainable solution is needed. This study used qualitative and quantitative methods to design, implement, and pilot a novel sanitation system in Kakuma refugee camp, Kenya. An initial round of 12 pre-implementation focus group discussions (FGDs) were conducted with Dinka and Somali residents to understand sanitation practices, perceptions, and needs. FGDs and a supplementary pre-implementation survey informed the development of an innovative sanitation management system that incorporated the provision of urine and liquid-diverting toilets, which separate urine and fecal waste, and a service-based sanitation system that included weekly waste collection. The new system was implemented on a pilot scale for 6 weeks. During the implementation, bi-weekly surveys were administered in each study household to monitor user perceptions and challenges. At the end of the pilot, the sanitation system was assessed using a second round of four post-implementation FGDs. Those who piloted the new sanitation system reported high levels of user satisfaction. Reported benefits included odor reduction, insect/pest reduction, the sitting design, the appropriateness for special populations, and waste collection. However, urine and liquid diversion presented a challenge for users who perform anal washing and for women who had experienced female genital mutilation. Refugee populations are often culturally and ethnically diverse. Using residents' input to inform the development of sanitation solutions can increase user acceptability and provide opportunities to improve sanitation system designs based on specific needs.


Present and potential therapy for allergic rhinitis: a review.
Allergic rhinitis can affect up to one-fifth of the population and the economic impact is increasing. H(1) receptor antagonists were the first major pharmacologic treatment, but the associated sedation limited their use. The 2 initial second generation less sedating antihistamines, astemizole and terfenadine, were found to prolong the cardiac QT(c) interval, especially when administered with other medications metabolised by the same cytochrome (CYP) P450 isoenzyme, CYP3A4. Other second generation antihistamines, fexofenadine, loratadine and cetirizine, do not cause clinically significant cardiac QT(c) interval prolongation. Two newer agents, ebastine and mizolastine, are also effective in the treatment of allergic rhinitis. Ebastine, however, prolongs the cardiac QT(c) interval in laboratory animals and humans, the clinical significance of which is unknown. Desloratadine and norastemizole, metabolites of loratadine and astemizole, respectively, are 2 other second generation antihistamines found to be effective treatments for seasonal allergic rhinitis. Unlike their parent compounds, they do not prolong the cardiac QT(c) interval. All clinically available intranasal corticosteroids are effective in the treatment of allergic rhinitis, but studies to evaluate possible long term systemic adverse effects are limited. Mometasone furoate and fluticasone propionate have lower oral bioavailability compared with other corticosteroids that are given intranasally. This may be important, since it is likely that some of the intranasal corticosteroid is ingested. Two 1-year growth studies in children indicated that intranasal beclomethasone dipropionate given twice daily reduces growth velocity, whereas intranasal mometasone furoate given once daily in the morning does not. Other studies are needed. Most but not all studies have shown that leukotriene antagonists are effective in the treatment of allergic rhinitis. H(1) receptor antagonists are not very effective in reducing nasal congestion, but leukotriene antagonists do attenuate this symptom. Furthermore, one study demonstrates an additive benefit in treating allergic rhinitis with the combination of a H(1) receptor and leukotriene antagonist. Clinical trials have demonstrated that anti-immunoglobulin (Ig) E is effective in the treatment of seasonal allergic rhinitis when free IgE is reduced to <25 microg/L. The reduction of total IgE is dose dependent and subcutaneous and intravenous administration are both effective. Immunotherapy is also an effective treatment for allergic rhinitis. CpG oligonucleotides is a novel adjuvant for allergen immunotherapy. This adjuvant used in a murine model shifts the immune response away from the allergic or TH2 phenotype. Studies in humans have not been performed.


Detailed Molecular and Immune Marker Profiling of Archival Prostate Cancer Samples Reveals an Inverse Association between TMPRSS2:ERG Fusion Status and Immune Cell Infiltration.
Prostate cancer is a significant global health issue, and limitations to current patient management pathways often result in overtreatment or undertreatment. New ways to stratify patients are urgently needed. We conducted a feasibility study of such novel assessments, looking for associations between genomic changes and lymphocyte infiltration. An innovative workflow using an in-house targeted sequencing panel, immune cell profiling using an image analysis pipeline, RNA sequencing, and exome sequencing in select cases was tested. Gene fusions were profiled by RNA sequencing in 27 of 27 cases, and a significantly higher tumor-infiltrating lymphocyte (TIL) count was noted in tumors without a TMPRSS2:ERG fusion compared with those with the fusion (P = 0.01). Although this finding was not replicated in a larger validation set (n = 436) of The Cancer Genome Atlas images, there was a trend in the same direction. Differential expression analysis of TIL-high and TIL-low tumors revealed the enrichment of both innate and adaptive immune response pathways. Mutations in mismatch repair genes (MLH1 and MSH6 mutations in 1 of 27 cases) were identified. We describe a potential immune escape mechanism in TMPRSS2:ERG fusion-positive tumors. Detailed profiling, as shown herein, can provide novel insights into tumor biology. Likely differences with findings with other cohorts are related to methods used to define region of interest, but this warrants further study in a larger cohort.


Salivary testosterone levels and autism-spectrum quotient in adults.
The purpose of this study is to examine the relationship between salivary testosterone levels and autistic traits in adults. A total of 92 male and female adults participated in the present study. Their salivary testosterone level (T) and score of Japanese version of Autism-spectrum Quotient (AQ) were assessed to examine the relationship between salivary testosterone level and autistic traits in adults. We observed a positive correlation between T and AQ in a group of both sexes. The correlation disappeared when we conducted correlation analysis by sex. However, although there was no sex difference in the score of the subscale of attention switching, attention switching was related to T. Although the relationship between T and AQ may mainly result from sex differences, the subscale of attention switching may be modulated by testosterone.


Nature-based solutions for hydro-meteorological hazards: Revised concepts, classification schemes and databases.
Hydro-meteorological hazards (HMHs) have had a strong impact on human societies and ecosystems. Their impact is projected to be exacerbated by future climate scenarios. HMHs cataloguing is an effective tool to evaluate their associated risks and plan appropriate remediation strategies. However, factors linked to HMHs origin and triggers remain uncertain, which pose a challenge for their cataloguing. Focusing on key HMHs (floods, storm surges, landslides, droughts, and heatwaves), the goal of this review paper is to analyse and present a classification scheme, key features, and elements for designing nature-based solutions (NBS) and mitigating the adverse impacts of HMHs in Europe. For this purpose, we systematically examined the literature on NBS classification and assessed the gaps that hinder the widespread uptake of NBS. Furthermore, we critically evaluated the existing literature to give a better understanding of the HMHs drivers and their interrelationship (causing multi-hazards). Further conceptualisation of classification scheme and categories of NBS shows that relatively few studies have been carried out on utilising the broader concepts of NBS in tackling HMHs and that the classification and effectiveness of each NBS are dependent on the location, architecture, typology, green species and environmental conditions, as well as interrelated non-linear systems. NBS are often more cost-effective than hard engineering approaches used within the existing systems, especially when taking into consideration their potential co-benefits. We also evaluated the sources of available data for HMHs and NBS, highlighted gaps in data, and presented strategies to overcome the current shortcomings for the development of the NBS for HMHs. We highlighted specific gaps and barriers that need to be filled since the uptake and upscaling studies of NBS in HMHs reduction is rare. The fundamental concepts and the key technical features of past studies reviewed here could help practitioners to design and implement NBS in a real-world situation.


The Effect of Preceding Race Efforts on Pacing and Short-Track Speed Skating Performance.
To examine whether preceding high-intensity race efforts in a competitive weekend affect pacing behavior and performance in elite short-track speed skaters. Finishing and intermediate lap times were gathered from 500-, 1000-, and 1500-m short-track speed skating world cups during the seasons 2011-2016. The effect of preceding races on pacing behavior and performance was explored using 2 studies. Study I: The effect of competing in extra races due to the repechage (Rep) system, leading to an increased number of high-intensity race efforts prior to the subsequent main tournament race, was explored (500-m, n = 32; 1000-m, n = 34; and 1500-m, n = 47). Study II: The performance of skaters over the tournament days was evaluated (500-m, n = 129; 1000-m, n = 54; and 1500-m, n = 114). For both analytic approaches, a 2-way repeated-measures analysis of variance was used to assess differences in pacing and performance within skaters over the races. An additional number of preceding high-intensity race efforts due to the Rep system reduced the qualification percentage in the first main tournament race for the next stage of competition in all events (500-m, direct qualification = 57.3%, Rep = 25.0%; 1000-m, direct = 44.2%, Rep = 28.3%; and 1500-m, direct = 27.1%, Rep = 18.2%) and led to a decreased pace in the initial 2 laps of the 500-m event. By contrast, tournament day (Saturday vs Sunday) only affected the pacing behavior of female skaters during the 1500-m event. High-intensity race efforts earlier in the day affected pacing and performance of elite skaters, whereas the effect of high-intensity race efforts from the previous day seemed to be only marginal.


Genome Wide Methylome Alterations in Lung Cancer.
Aberrant cytosine 5-methylation underlies many deregulated elements of cancer. Among paired non-small cell lung cancers (NSCLC), we sought to profile DNA 5-methyl-cytosine features which may underlie genome-wide deregulation. In one of the more dense interrogations of the methylome, we sampled 1.2 million CpG sites from twenty-four NSCLC tumor (T)-non-tumor (NT) pairs using a methylation-sensitive restriction enzyme- based HELP-microarray assay. We found 225,350 differentially methylated (DM) sites in adenocarcinomas versus adjacent non-tumor tissue that vary in frequency across genomic compartment, particularly notable in gene bodies (GB; p<2.2E-16). Further, when DM was coupled to differential transcriptome (DE) in the same samples, 37,056 differential loci in adenocarcinoma emerged. Approximately 90% of the DM-DE relationships were non-canonical; for example, promoter DM associated with DE in the same direction. Of the canonical changes noted, promoter (PR) DM loci with reciprocal changes in expression in adenocarcinomas included HBEGF, AGER, PTPRM, DPT, CST1, MELK; DM GB loci with concordant changes in expression included FOXM1, FERMT1, SLC7A5, and FAP genes. IPA analyses showed adenocarcinoma-specific promoter DMxDE overlay identified familiar lung cancer nodes [tP53, Akt] as well as less familiar nodes [HBEGF, NQO1, GRK5, VWF, HPGD, CDH5, CTNNAL1, PTPN13, DACH1, SMAD6, LAMA3, AR]. The unique findings from this study include the discovery of numerous candidate The unique findings from this study include the discovery of numerous candidate methylation sites in both PR and GB regions not previously identified in NSCLC, and many non-canonical relationships to gene expression. These DNA methylation features could potentially be developed as risk or diagnostic biomarkers, or as candidate targets for newer methylation locus-targeted preventive or therapeutic agents.


Structure and enzymatic properties of genetically truncated forms of the water-insoluble glucan-synthesizing glucosyltransferase from Streptococcus sobrinus.
Glucosyltransferase-I (GTF-I: 175 kDa) of a cariogenic bacterium, Streptococcus sobrinus 6715, mediates the conversion of water-soluble dextran (alpha-1,6-glucan) into a water-insoluble form by making numerous alpha-1,3-glucan branches along the dextran chains with sucrose as the glucosyl donor. The structures and catalytic properties were compared for two GTF-I fragments, GTF-I' (138 kDa) and GS (110 kDa). Both lack the N-terminal 84 residues of GTF-I. While GTF-I' still contains four of the six C-terminal repeats characteristic of streptococcal glucosyltransferases, GS lacks all of them. Electron microscopy of negatively stained samples indicated a double-domain structure for GTF-I', consisting of a spherical head with a smaller spherical tail, which was occasionally seen as a long extension. GS was seen just as the head portion of GTF-I'. In the absence of dextran, both fragments simply hydrolyzed sucrose with similar K(m) and k(cat) values at low concentrations (<5 mM). At higher sucrose concentrations (>10 mM), however, GTF-I' exhibited glucosyl transfer activity to form insoluble alpha-1, 3-glucans. So did GS, but less efficiently. Dextran increased the rate and efficiency of the glucosyl transfer by GTF-I'. On removal of the C-terminal repeats of GTF-I' by mild trypsin treatment, this dextran-stimulated transfer was completely lost and the dextran-independent transfer became less efficient. These results indicate that the N-terminal two-thirds of the GTF-I sequence are organized as a structurally and functionally independent domain to catalyze not only sucrose hydrolysis but also glucosyl transfer to form alpha-1,3-glucan chains, although not efficiently; the C-terminal repeat increases the efficiency of the intrinsic glucosyl transfer by the N-terminal domain as well as rendering the whole molecule primer-dependent for far more efficient insoluble glucan synthesis.


Validity of the clinical dementia rating scale for the detection and staging of dementia in Brazilian patients.
The aim of this study was to determine the diagnostic value and agreement analyses between Clinical Dementia Rating (CDR) and dementia diagnostic criteria (gold standard), Blessed Dementia Rating scale (BDRS), and Diagnostic and Statistical Manual of Mental Disorders, 3rd Edition, Revised (DSM III-R) criteria for severity. In a sample of 343 Southern Brazilian participants, CDR was consecutively assessed in 295 dementia patients (Alzheimer disease, vascular dementia, and questionable) and 48 healthy elderly. The National Institute of Neurological and Communicative Diseases and Stroke/Alzheimer's Disease and Related Disorders Association (NINCDS-ADRDA) criteria for probable Alzheimer disease and the National Institute of Neurological Disorders and Stroke and Association Internationale pour la Recherché et l'Enseignement en Neurosciences (NINDS-AIREN) for probable vascular dementia were the gold standard. A battery of cognitive tests and the Mini Mental State Examination (as a screening test at study entry) were also applied. Sensitivity and specificity were obtained through contingency tables. Validity and reliability were measured through kappa coefficient, Kendall b, and percent agreement. CDR agreement among raters was demonstrated by percent agreement. Agreement to gold standard was good (kappa=0.75), as well as to the Blessed scale (kappa=0.73), and excellent to the DSM III-R (kappa=0.78). CDR detection of dementia among healthy elderly or questionable dementia was 86% and 80% sensitive, respectively, and 100% specific for both settings. In conclusion, agreement of CDR global score with the gold standard was good, and diagnostic values were high.


Simultaneous quantitation of the highly lipophilic atovaquone and hydrophilic strong basic proguanil and its metabolites using a new mixed-mode SPE approach and steep-gradient LC.
A bioanalytical method is described for the simultaneous quantitative analysis of the highly lipophilic atovaquone and the strong basic proguanil with metabolites in plasma. The drugs are extracted from protein precipitated plasma samples on a novel mixed-mode solid-phase extraction (SPE) column containing carboxypropyl and octyl silica as functional groups. The analytes are further separated and quantitated using a steep-gradient liquid chromatographic method on a Zorbax SB-CN column with UV detection at 245 nm. Two different internal standards (IS) are used in the method to compensate for both types of analytes. A structurally similar IS to atovaquone is added with acetonitrile to precipitate proteins from plasma. A structurally similar IS to proguanil and its metabolites is added with phosphate buffer before samples are loaded onto the SPE columns. A single elution step is sufficient to elute all analytes. The method is validated according to published guidelines and shows excellent performance. The within-day precisions, expressed as relative standard deviation, are lower than 5% for all analytes at three tested concentrations within the calibration range. The between-day precisions are lower than 13% for all analytes at the same tested concentrations. The limit of quantitation is 25 nM for the basic substances and 50 nM for atovaquone. Several considerations regarding development and optimization of a method for determination of analytes with such a difference in physiochemical properties are discussed.


Imageability ratings across languages.
Imageability is a psycholinguistic variable that indicates how well a word gives rise to a mental image or sensory experience. Imageability ratings are used extensively in psycholinguistic, neuropsychological, and aphasiological studies. However, little formal knowledge exists about whether and how these ratings are associated between and within languages. Fifteen imageability databases were cross-correlated using nonparametric statistics. Some of these corresponded to unpublished data collected within a European research network-the Collaboration of Aphasia Trialists (COST IS1208). All but four of the correlations were significant. The average strength of the correlations (rho = .68) and the variance explained (R 2 = 46%) were moderate. This implies that factors other than imageability may explain 54% of the results. Imageability ratings often correlate across languages. Different possibly interacting factors may explain the moderate strength and variance explained in the correlations: (1) linguistic and cultural factors; (2) intrinsic differences between the databases; (3) range effects; (4) small numbers of words in each database, equivalent words, and participants; and (5) mean age of the participants. The results suggest that imageability ratings may be used cross-linguistically. However, further understanding of the factors explaining the variance in the correlations will be needed before research and practical recommendations can be made.