Antagonistic activities of cotranscriptional regulators within an early developmental window set FLC expression level.
Quantitative variation in expression of the Arabidopsis floral repressor FLC influences whether plants overwinter before flowering, or have a rapid cycling habit enabling multiple generations a year. Genetic analysis has identified activators and repressors of FLC expression but how they interact to set expression level is poorly understood. Here, we show that antagonistic functions of the FLC activator FRIGIDA (FRI) and the repressor FCA, at a specific stage of embryo development, determine FLC expression and flowering. FRI antagonizes an FCA-induced proximal polyadenylation to increase FLC expression and delay flowering. Sector analysis shows that FRI activity during the early heart stage of embryo development maximally delays flowering. Opposing functions of cotranscriptional regulators during an early embryonic developmental window thus set FLC expression levels and determine flowering time.


The in vivo effects of the Pro12Ala PPARgamma2 polymorphism on adipose tissue NEFA metabolism: the first use of the Oxford Biobank.
To investigate the phenotypic effects of common polymorphisms on adipose tissue metabolism and cardiovascular risk factors, we set out to establish a biobank with the unique feature of allowing a prospective recruit-by-genotype approach. The first use of this biobank investigates the effects of the peroxisome proliferator-activated receptor (PPAR) Pro12Ala polymorphism on integrative tissue-specific physiology. We hypothesised that Ala12 allele carriers demonstrate greater adipose tissue metabolic flexibility and insulin sensitivity. From a comprehensive population register, subjects were recruited into a biobank, which was genotyped for the Pro12Ala polymorphism. Twelve healthy male Ala12 carriers and 12 matched Pro12 homozygotes underwent detailed physiological phenotyping using stable isotope techniques, and measurements of blood flow and arteriovenous differences in adipose tissue and muscle in response to a mixed meal containing [1,1,1-(13)C]tripalmitin. Of 6,148 invited subjects, 1,072 were suitable for inclusion in the biobank. Among Pro12 homozygotes, insulin sensitivity correlated with HDL-cholesterol concentrations, and inversely correlated with blood pressure, apolipoprotein B, triglyceride and total cholesterol concentrations. Ala12 carriers showed no such correlations. In the meal study, Ala12 carriers had lower plasma NEFA concentrations, higher adipose tissue and muscle blood flow, and greater insulin-mediated postprandial hormone-sensitive lipase suppression along with greater insulin sensitivity than Pro12 homozygotes. This study shows that a recruit-by-genotype approach is feasible and describes the biobank's first application, providing tissue-specific physiological findings consistent with the epidemiological observation that the PPAR Ala12 allele protects against the development of type 2 diabetes.


Exosomal CCL2 from Tubular Epithelial Cells Is Critical for Albumin-Induced Tubulointerstitial Inflammation.
Albuminuria is a key instigator of tubulointerstitial inflammation associated with CKD, but the mechanism through which filtered albumin propagates renal injury remains unclear. In this study, we explored the role in this process of exosome mRNA released from tubular epithelial cells (TECs). Compared with control mice, acute and chronic kidney injury models had more exosomes containing inflammatory cytokine mRNA, particularly the chemokine CCL2, in kidneys and urine. In vitro stimulation of TECs with BSA recapitulated this finding. Notably, the internalization of purified TEC exosomes by cultured macrophages increased if TECs were exposed to BSA. Macrophage internalization of exosomes from BSA-treated TECs led to an enhanced inflammatory response and macrophage migration, but CCL2 silencing in TECs prevented these effects. Using a GFP-CCL2 fusion mRNA construct, we observed direct transfer of CCL2 mRNA from TEC exosomes to macrophages. Mice subjected to tail vein injection of purified BSA-treated TEC exosomes developed tubular injury with renal inflammatory cell infiltration. However, injection of exosomes from BSA-treated CCL2-deficient TECs induced less severe kidney inflammation. Finally, in patients with IgA nephropathy, the increase of proteinuria correlated with augmented urinary excretion of exosomes with exaggerated expression of CCL2 mRNA. Moreover, the level of CCL2 mRNA in urinary exosomes correlated closely with levels of renal interstitial macrophage infiltration in these patients. Our studies demonstrate that the increasing release of exosomes that transfer CCL2 mRNA from TECs to macrophages constitutes a critical mechanism of albumin-induced tubulointerstitial inflammation.


Insights into the mechanisms of catalysis and heterotropic regulation of Escherichia coli aspartate transcarbamoylase based upon a structure of the enzyme complexed with the bisubstrate analogue N-phosphonacetyl-L-aspartate at 2.1 A.
A high-resolution structure of Escherichia coli aspartate transcarbamoylase has been determined to 2.1 A; resolution in the presence of the bisubstrate analog N-phosphonacetyl-L-aspartate (PALA). The structure was refined to a free R-factor of 23.4% and a working R-factor of 20.3%. The PALA molecule is completely saturated with interactions to side chain and backbone groups in the active site, including two interactions that are contributed from the 80s loop of the adjacent catalytic chain. The charge neutralization of the bound PALA molecule (and presumably the substrates as well) induced by the electrostatic field of the highly positively charged active site is an important factor in the high binding affinity of PALA and must be important for catalysis. The higher-resolution structure reported here departs in a number of ways from the previously determined structure at lower resolution. These modifications include alterations in the backbone conformation of the C-terminal of the catalytic chains, the N- and C-termini of the regulatory chains, and two loops of the regulatory chain. The high-resolution of this structure has allowed a more detailed description of the binding of PALA to the active site of the enzyme and has allowed a detailed model of the tetrahedral intermediate to be constructed. This model becomes the basis of a description of the catalytic mechanism of the transcarbamoylase reaction. The R-structural state of the enzyme-PALA complex is an excellent representation of the form of the enzyme that occurs at the moment in the catalytic cycle when the tetrahedral intermediate is formed. Finally, improved electron density in the N-terminal region of the regulatory chain (residues 1 to 7) has allowed tracing of the entire regulatory chain. The N-terminal segments of the R1 and R6 chains are located in close proximity to each other and to the regulatory site. This portion of the molecule may be involved in the observed asymmetry between the regulatory binding sites as well as in the heterotropic response of the enzyme.


Bombus impatiens (Hymenoptera: Apidae): an alternative to Apis mellifera (Hymenoptera: Apidae) for lowbush blueberry pollination.
The pollination effectiveness of the commercially reared bumble bee Bombus impatiens Cresson, was compared in field studies to the honey bee, Apis mellifera L., for lowbush blueberry, Vaccinium angustifolium Ait. A preliminary study indicated that B. impatiens had potential as an alternative pollinator. In a 3-yr study, percentage fruit set, percentage harvested berries, berry weight, and seeds per berry were compared in blueberry fields stocked at 7.5 A. mellifera hives per hectare to 5, 7.5, or 10 B. impatiens colonies per hectare. Percentage of harvested berries (yield) was significantly higher in fields stocked with B. impatiens at 10 colonies per hectare. No other parameters measuring pollinator effectiveness were significantly different at 5, 7.5, or 10 colonies per hectare. Flower handling time was significantly faster for B. impatiens and it more frequently collected blueberry pollen. All parameters of pollinator effectiveness were similar for B. impatiens, A. mellifera, and native wild bees in a follow-up study. Overall, B. impatiens was a suitable alternative to A. mellifera.


Trypanosoma cruzi infection in offspring born to chagasic C3H/He mice.
This study reports the effects of Trypanosoma cruzi infection induced in C3H/He male and female mice born to chagasic mice. An experimental model was established infecting female C3H/He mice with a low virulent T. cruzi clone. In this model, mating, fertilization, pregnancy evolution and delivery was carried out successfully. The offspring was infected at four, six and eighth weeks of age. The results showed that the offspring born to chagasic mothers present decreased resistance to acquired T. cruzi infection. This decreased resistance was expressed by higher levels of parasitaemia and higher mortality rates in offspring born to chagasic mothers than in controls. Age and sex were shown to be important factors of this phenomenon. The results suggest that maternal immune system products can modulate the immune response of the offspring.


Heterocyclic aromatic amine intake increases colorectal adenoma risk: findings from a prospective European cohort study.
Heterocyclic aromatic amines (HCAs), which arise from cooking meat and fish at high temperatures, may increase the risk of colorectal adenomas. Conversely, flavonoids might counteract the negative effects of HCAs. The association between dietary HCA intake and colorectal adenoma incidence was investigated in a prospective cohort study. At recruitment (1994-1998), detailed information on diet, anthropometric measures, lifestyle, and medication use was assessed in 25,540 participants of the European Prospective Investigation into Cancer and Nutrition-Heidelberg cohort study. Dietary HCA intake was estimated by using information from food-frequency questionnaires on meat consumption, applied cooking methods, and preferred degree of browning. Until June 2007, 516 verified incident colorectal adenomas were identified. Participants with negative colonoscopy (n = 3966) were also included in the analytic cohort. Multivariate Cox proportional hazards regression was used to examine the association between colorectal adenoma risk and intake of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP), 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx), and 2-amino-3,4,8-dimethylimidazo[4,5-f]quinoxaline (DiMeIQx). In multivariate analyses, the intake of PhIP as the most abundant dietary HCA was associated with an increased risk of colorectal adenoma (relative risk: 1.47; 95% CI: 1.13, 1.93; quartile 4 compared with quartile 1; P for trend = 0.002), but no statistically significant associations were observed for MeIQx and DiMeIQx intakes. In addition, adenoma risk also increased with the consumption of strongly or extremely browned meat (P for trend = 0.04). The association of PhIP intake with adenoma risk was most pronounced for small adenomas (P for trend = 0.01) and adenomas localized in the distal colon (P for trend = 0.002). The results of this first European cohort study support data from case-control studies of a positive association between HCA intake and colorectal adenoma risk.


Cyberbullying victimization and its association with health across the life course: A Canadian population study.
To examine the prevalence of cyberbullying victimization (CV), its associations with self-reported health and substance use and the extent to which age moderates these associations. We used the 2014 Canadian General Social Survey on Victimization (N = 31 907, mean age = 45.83, SD = 18.67) and binary logistic regression models to estimate the strength of association between CV and health-related outcomes. The five-year prevalence of CV was 5.1%. Adolescents reported the highest prevalence of CV (12.2%), compared to all other adult age groups (1.7%-10.4%). After controlling for socio-demographic covariates, individuals exposed to CV had increased odds of reporting poor mental health (OR = 4.259, 95% CI = 2.853-6.356), everyday limitations due to mental health problems (OR = 3.263, 95% CI = 2.271-4.688), binge drinking (OR = 2.897, 95% CI = 1.765-4.754), and drug use (OR = 3.348, 95% CI = 2.333-4.804), compared to those not exposed to CV. The associations between CV and self-reported mental health and substance use were strongest for adolescents and attenuated across the adult age groups. Adolescence may represent a developmental period of heightened susceptibility to CV. Developing and evaluating targeted preventive interventions for this age group is warranted.


Identifying the Tuskegee Syphilis Study: implications of results from recall and recognition questions.
This analysis assessed whether Blacks, Whites and Puerto-Rican (PR) Hispanics differed in their ability to identify the Tuskegee Syphilis Study (TSS) via open-ended questions following lead-in recognition and recall questions. The Tuskegee Legacy Project (TLP) Questionnaire was administered via a Random-Digit Dial (RDD) telephone survey to a stratified random sample of Black, White and PR Hispanic adults in three U.S. cities. The TLP Questionnaire was administered to 1,162 adults (356 African-Americans, 313 PR Hispanics, and 493 non-Hispanic Whites) in San Juan, PR, Baltimore, MD and New York City, NY. Recall question data revealed: 1) that 89% or more of Blacks, Whites, and PR Hispanics were not able to name or definitely identify the Tuskegee Syphilis Study by giving study attributes; and, 2) that Blacks were the most likely to provide an open-ended answer that identified the Tuskegee Syphilis Study as compared to Whites and PR Hispanics (11.5% vs 6.3% vs 2.9%, respectively) (p <or= 0.002). Even when probed by a recognition question, only a minority of each racial/ethnic group (37.1%, 26.9%, and 8.6%, for Blacks, Whites and PR Hispanics, respectively) was able to clearly identify the TSS (p < 0.001). The two major implications of these findings for health disparity researchers are 1) that it is unlikely that detailed knowledge of the Tuskegee Syphilis Study has any current widespread influence on the willingness of minorities to participate in biomedical research, and 2) that caution should be applied before assuming that what community leaders 'know and are aware of' is equally 'well known' within their community constituencies.


Intrathecal, polyspecific antiviral immune response in oligoclonal band negative multiple sclerosis.
Oligoclonal bands (OCB) are detected in the cerebrospinal fluid (CSF) in more than 95% of patients with multiple sclerosis (MS) in the Western hemisphere. Here we evaluated the intrathecal, polyspecific antiviral immune response as a potential diagnostic CSF marker for OCB-negative MS patients. We tested 46 OCB-negative German patients with paraclinically well defined, definite MS. Sixteen OCB-negative patients with a clear diagnosis of other autoimmune CNS disorders and 37 neurological patients without evidence for autoimmune CNS inflammation served as control groups. Antibodies against measles, rubella, varicella zoster and herpes simplex virus in paired serum and CSF samples were determined by ELISA, and virus-specific immunoglobulin G antibody indices were calculated. An intrathecal antibody synthesis against at least one neurotropic virus was detected in 8 of 26 (31%) patients with relapsing-remitting MS, 8 of 12 (67%) with secondary progressive MS and 5 of 8 (63%) with primary progressive MS, in 3 of 16 (19%) CNS autoimmune and 3 of 37 (8%) non-autoimmune control patients. Antibody synthesis against two or more viruses was found in 11 of 46 (24%) MS patients but in neither of the two control groups. On average, MS patients with a positive antiviral immune response were older and had a longer disease duration than those without. Determination of the intrathecal, polyspecific antiviral immune response may allow to establish a CSF-supported diagnosis of MS in OCB-negative patients when two or more of the four virus antibody indices are elevated.


Self-reporting hybridisation assay for miRNA analysis.
Hybridisation assays, which are commonly used to analyse oligonucleotides such as siRNAs and miRNAs, often employ detection probes with fluorescent tags. The signal emitted by a fluorescent tag covers a broad range of wavelengths and this limits the multiplexing potential due to overlapping signals. A novel method of indirect oligonucleotide analysis has been developed which combines a hybridisation assay with cleavable small molecule mass tags using HPLC-ESI MS detection. A self-reporting detection probe has been designed which incorporates a DNA/RNA chimeric oligonucleotide sequence in the reporter region, which generates small nucleotide products upon RNase cleavage of the ribose-phosphate backbone. These small nucleotides can then serve as mass tags for the indirect detection of oligonucleotide analytes. The narrow mass range covered by a small molecule mass tag combined with the wide range of possible mass tags provides a high degree of multiplexing potential. This approach has been demonstrated for the analysis of a synthetic miRNA.


Effect of medium-chain glycerides on the intestinal absorption of phenol red: studies on mechanisms of the promoting effect.
To elucidate the mechanisms of the promoting action of medium-chain glyceride (MCG) on the intestinal absorption of phenol red (PR), the effect of different MCG dose levels on PR absorption, the absorption kinetics of MCG itself, and the mucosal reduced glutathione (GSH) levels were investigated to determine whether or not they are related to the promoting action of MCG. The absorption of PR was found to be enhanced with an increase in MCG dose (as an emulsion in the range 0.5-4.0%). Urinary excretion of PR was reduced by absorbed MCG components, which also resulted in an increase in the AUC. Moment analysis of plasma PR concentrations after intraintestinal administration revealed a shorter mean absorption time (MAT) for the emulsion formulation than for the control. The mucosal GSH level was lowered by the administration of the MCG emulsion (2-10%) into the intestinal lumen. Pretreatment with a 4% MCG emulsion decreased GSH levels to one-half of the control. In these cases, the level of mucosal GSH seemed to remain constant to the end of the experiment after it had decreased during the initial period. On the other hand, results from the in situ loop or recirculation study showed that enhancement of PR absorption appeared in the earlier period and then gradually decreased. The time course of the PR absorption rate, estimated from plasma concentration data, was similar to that of the MCG absorption rate. These results suggest that the promotion of intestinal absorption is related to the absorption kinetics of MCG itself, but not directly to the level of mucosal GSH.


Characterization of Penicillium Isolates Associated with Blue Mold on Apple in Uruguay.
Blue mold caused by Penicillium spp. is the most important postharvest disease of apple in Uruguay. Fourteen isolates of Penicillium were recovered from rotten apple and pear fruit with blue mold symptoms, and from water from flotation tanks in commercial apple juice facilities. Phenotypic identification to species level was performed, and the isolates were tested for sensitivity to commonly used postharvest fungicides. Genetic characterization of the isolates was performed with restriction fragment length polymorphism of the region including the internal transcribed spacer (ITS) ITS1 and ITS2 and the 5.8SrRNA gene (ITS1-5.8SrRNA gene-ITS2) ribosomal DNA region and with random amplified polymorphic DNA (RAPD) primers. Both techniques were able to differentiate these isolates at the species level. RAPD analysis proved to be an objective, rapid, and reliable tool to identify Penicillium spp. involved in blue mold of apple. In all, 11 isolates were identified as Penicillium expansum and 3 as P. solitum. This is the first report of P. solitum as an apple pathogen in Uruguay.


Integrin binding and mechanical tension induce movement of mRNA and ribosomes to focal adhesions.
The extracellular matrix (ECM) activates signalling pathways that control cell behaviour by binding to cell-surface integrin receptors and inducing the formation of focal adhesion complexes (FACs). In addition to clustered integrins, FACs contain proteins that mechanically couple the integrins to the cytoskeleton and to immobilized signal-transducing molecules. Cell adhesion to the ECM also induces a rapid increase in the translation of preexisting messenger RNAs. Gene expression can be controlled locally by targeting mRNAs to specialized cytoskeletal domains. Here we investigate whether cell binding to the ECM promotes formation of a cytoskeletal microcompartment specialized for translational control at the site of integrin binding. High-resolution in situ hybridization revealed that mRNA and ribosomes rapidly and specifically localized to FACs that form when cells bind to ECM-coated microbeads. Relocation of these protein synthesis components to the FAC depended on the ability of integrins to mechanically couple the ECM to the contractile cytoskeleton and on associated tension-moulding of the actin lattice. Our results suggest a new type of gene regulation by integrins and by mechanical stress which may involve translation of mRNAs into proteins near the sites of signal reception.


Effect of a gluten-free diet on gastrointestinal symptoms in celiac disease.
Atypical presentations of celiac disease appear to be at least as common as is the classic presentation of steatorrhea, diarrhea, and weight loss. We examined the effect of a gluten-free diet on gastrointestinal symptoms in a cohort of US patients with celiac disease. A follow-up survey was conducted in 215 patients who were evaluated at the University of Iowa from 1990 through 1997 as having biopsy-confirmed celiac disease. The systematic survey asked detailed questions regarding gastrointestinal symptoms before and after the institution of a gluten-free diet in the patients, all of whom had been given the same dietary advice. The group consisted of 160 female and 55 male patients. Although diarrhea was the most frequent symptom in untreated celiac disease, steatorrhea occurred in only one-fifth of patients. Other complaints were common, and most responded to gluten exclusion. The benefit of gluten exclusion was equally apparent in men and women. Diarrhea responded in most patients, usually within days, and the mean time to resolution was 4 wk. Many patients had alternating diarrhea and constipation, both of which were responsive to the gluten-free diet. Most patients had abdominal pain and bloating, which resolved with the diet. Celiac disease causes a wide range of gastrointestinal symptoms. Clinicians must have a high level of suspicion to detect the atypical forms of celiac disease. With a gluten-free diet, patients have substantial and rapid improvement of symptoms, including symptoms other than the typical ones of diarrhea, steatorrhea, and weight loss.


Chinese hamster ovary cell mutants resistant to DNA polymerase inhibitors. I. Isolation and biochemical genetic characterization.
Isolation and characterization of Chinese hamster ovary cell mutants resistant to different DNA polymerase inhibitors (aphidicolin, ara-A and ara-C) have been described. A particular mutant (JK3-1-2A) characterized in detail was found to grow and synthesize DNA in medium containing an amount of aphidicolin tenfold greater than that which completely inhibited the growth and the DNA synthesis of the wild-type cells. An almost twofold increase in the specific activity of the DNA polymerase alpha was seen in this mutant. The mutant DNA polymerase showed altered aphidicolin inhibition kinetics of dCMP incorporation; the apparent Km for dCTP and the apparent Ki for aphidicolin were increased in the mutant. These alterations in the kinetic parameters were, however, abolished upon further purification of the enzyme. Ara-CTP was found to act as a competitive inhibitor of the dCMP incorporation by both the wild type and mutant enzymes. In contrast, the effect of aphidicolin on dCMP incorporation was either competitive (wild-type enzymes) or noncompetitive (mutant enzyme). The data presented showed that the sites of action for aphidicolin and ara-CTP were distinct; likewise the dCTP binding site appeared to be separate from other dNTP(s) binding sites. The drug resistance of the mutant was inherited as a dominant trait.


The impact of nurse working hours on patient safety culture: a cross-national survey including Japan, the United States and Chinese Taiwan using the Hospital Survey on Patient Safety Culture.
A positive patient safety culture (PSC) is one of the most critical components to improve healthcare quality and safety. The Hospital Survey on Patient Safety Culture (HSOPS), developed by the US Agency for Healthcare Research and Quality, has been used to assess PSC in 31 countries. However, little is known about the impact of nurse working hours on PSC. We hypothesized that long nurse working hours would deteriorate PSC, and that the deterioration patterns would vary between countries. Moreover, the common trends observed in Japan, the US and Chinese Taiwan may be useful to improve PSC in other countries. The purpose of this study was to clarify the impact of long nurse working hours on PSC in Japan, the US, and Chinese Taiwan using HSOPS. The HSOPS questionnaire measures 12 sub-dimensions of PSC, with higher scores indicating a more positive PSC. Odds ratios (ORs) were calculated using a generalized linear mixed model to evaluate the impact of working hours on PSC outcome measures (patient safety grade and number of events reported). Tukey's test and Cohen's d values were used to verify the relationships between nurse working hours and the 12 sub-dimensions of PSC. Nurses working ≥60 h/week in Japan and the US had a significantly lower OR for patient safety grade than those working <40 h/week. In the three countries, nurses working ≥40 h/week had a significantly higher OR for the number of events reported. The mean score on 'staffing' was significantly lower in the ≥60-h group than in the <40-h group in all the three countries. The mean score for 'teamwork within units' was significantly lower in the ≥60-h group than in the <40-h group in Japan and Chinese Taiwan. Patient safety grade deteriorated and the number of events reported increased with long working hours. Among the 12 sub-dimensions of PSC, long working hours had an impact on 'staffing' and 'teamwork within units' in Japan, the US and Chinese Taiwan.


Murine T Cell Maturation Entails Protection from MBL2, but Complement Proteins Do Not Drive Clearance of Cells That Fail Maturation in the Absence of NKAP.
Recent thymic emigrants that fail postpositive selection maturation are targeted by complement proteins. T cells likely acquire complement resistance during maturation in the thymus, a complement-privileged organ. To test this, thymocytes and fresh serum were separately obtained and incubated together in vitro to assess complement deposition. Complement binding decreased with development and maturation. Complement binding decreased from the double-positive thymocyte to the single-positive stage, and within single-positive thymocytes, complement binding gradually decreased with increasing intrathymic maturation. Binding of the central complement protein C3 to wild-type immature thymocytes required the lectin but not the classical pathway. Specifically, MBL2 but not MBL1 was required, demonstrating a unique function for MBL2. Previous studies demonstrated that the loss of NKAP, a transcriptional regulator of T cell maturation, caused peripheral T cell lymphopenia and enhanced complement susceptibility. To determine whether complement causes NKAP-deficient T cell disappearance, both the lectin and classical pathways were genetically ablated. This blocked C3 deposition on NKAP-deficient T cells but failed to restore normal cellularity, indicating that complement contributes to clearance but is not the primary cause of peripheral T cell lymphopenia. Rather, the accumulation of lipid peroxides in NKAP-deficient T cells was observed. Lipid peroxidation is a salient feature of ferroptosis, an iron-dependent nonapoptotic cell death. Thus, wild-type thymocytes naturally acquire the ability to protect themselves from complement targeting by MBL2 with maturation. However, NKAP-deficient immature peripheral T cells remain scarce in complement-deficient mice likely due to ferroptosis.


Geographic parthenogenesis and plant-enemy interactions in the common dandelion.
Many species with sexual and asexual variants show a pattern of geographic parthenogenesis where asexuals have broader and higher-latitude distribution than sexuals. Because sexual reproduction is often considered a costly evolutionary strategy that is advantageous in the face of selection by coevolving pests and pathogens, one possible explanation for geographic parthenogenesis is that populations at higher latitudes are exposed to fewer pests and pathogens. We tested this hypothesis in the common dandelion (Taraxacum officinale), a species with well-established geographic parthenogenesis, by screening prevalence and effects of several specialized pests and pathogens in natural dandelion populations. We did a population survey of 18 dandelion populations along a geographic transect that ranged from the area where sexual and asexual dandelions co-occur northward into the area where only asexuals occur. In addition we used four southern and four northern populations in a 8x8 cross-inoculation greenhouse experiment in which plants were exposed experimentally to each other's natural field soil microbial communities. The cross-inoculation experiment indicated a higher pathogenicity of soil microbial communities from the southern, mostly sexual, populations compared to soil microbial communities from the northern asexual populations. Northern dandelion populations also showed reduced infestation by a specialized seed-eating weevil. A similar trend of reduced rust fungus infection in northern populations was observed but this trend was not statistically significant. The prevalence of pests and pathogens decreased along the south-to-north axis of geographic parthenogenesis. This highlights the potential of biotic interactions in shaping patterns of geographic parthenogenesis.


Effectiveness of bladder tumor antigen quantitative test in the diagnosis of bladder carcinoma in a schistosoma endemic area.
Bladder carcinoma is the most common male cancer in our environment due to endemicity of schistosomiasis. Squamous-cell carcinoma is the most common histological type and patients present at an advanced stage. The objective of this study is to compare the sensitivity, specificity, and predictive values of the bladder tumor antigen quantitative test (BTA TRAK) and urine cytology in the diagnosis of bladder carcinoma in a schistosoma endemic area. This is a 12-month cross-sectional study of 88 patients, 52 of them with features of bladder carcinoma as study group, and 36 of them with hematuria from other urologic conditions, and benign urologic conditions and healthy volunteers as control group (CG). The mean ages of patients in the study and CGs were 47.17 ± 17.00 and 44.19 ± 18.89 years, respectively (P = 0.412). Bladder tumor antigen was assayed using enzyme-linked immunosorbent assay. Data were analyzed using SPSS version 20.0 for Windows. The sensitivity of urine cytology and BTA TRAK in the study was 29.1% and 98.8%, respectively. The specificity of urine cytology and BTA TRAK was 95.5% and 13.6%, respectively (P = 0.05). The positive predictive values of urine cytology and BTA TRAK in the study were 96.2% and 81.7%, respectively. The negative predictive values were 25.0% and 75.0% for urine cytology and BTA TRAK, respectively. BTA TRAK is more sensitive but poorly specific as compared to that of the urine cytology for bladder cell carcinoma detection in a schistosoma endemic area.


Epigenome-wide association study of serum urate reveals insights into urate co-regulation and the SLC2A9 locus.
Elevated serum urate levels, a complex trait and major risk factor for incident gout, are correlated with cardiometabolic traits via incompletely understood mechanisms. DNA methylation in whole blood captures genetic and environmental influences and is assessed in transethnic meta-analysis of epigenome-wide association studies (EWAS) of serum urate (discovery, n = 12,474, replication, n = 5522). The 100 replicated, epigenome-wide significant (p < 1.1E-7) CpGs explain 11.6% of the serum urate variance. At SLC2A9, the serum urate locus with the largest effect in genome-wide association studies (GWAS), five CpGs are associated with SLC2A9 gene expression. Four CpGs at SLC2A9 have significant causal effects on serum urate levels and/or gout, and two of these partly mediate the effects of urate-associated GWAS variants. In other genes, including SLC7A11 and PHGDH, 17 urate-associated CpGs are associated with conditions defining metabolic syndrome, suggesting that these CpGs may represent a blood DNA methylation signature of cardiometabolic risk factors. This study demonstrates that EWAS can provide new insights into GWAS loci and the correlation of serum urate with other complex traits.


Structural brain abnormalities in postural tachycardia syndrome: A VBM-DARTEL study.
Postural tachycardia syndrome (PoTS), a form of dysautonomia, is characterized by orthostatic intolerance, and is frequently accompanied by a range of symptoms including palpitations, lightheadedness, clouding of thought, blurred vision, fatigue, anxiety, and depression. Although the estimated prevalence of PoTS is approximately 5-10 times as common as the better-known condition orthostatic hypotension, the neural substrates of the syndrome are poorly characterized. In the present study, we used magnetic resonance imaging (MRI) with voxel-based morphometry (VBM) applying the diffeomorphic anatomical registration through exponentiated lie algebra (DARTEL) procedure to examine variation in regional brain structure associated with PoTS. We recruited 11 patients with established PoTS and 23 age-matched normal controls. Group comparison of gray matter volume revealed diminished gray matter volume within the left anterior insula, right middle frontal gyrus and right cingulate gyrus in the PoTS group. We also observed lower white matter volume beneath the precentral gyrus and paracentral lobule, right pre- and post-central gyrus, paracentral lobule and superior frontal gyrus in PoTS patients. Subsequent ROI analyses revealed significant negative correlations between left insula volume and trait anxiety and depression scores. Together, these findings of structural differences, particularly within insular and cingulate components of the salience network, suggest a link between dysregulated physiological reactions arising from compromised central autonomic control (and interoceptive representation) and increased vulnerability to psychiatric symptoms in PoTS patients.


Prediction of transmission distortion for wireless video communication: analysis.
Transmitting video over wireless is a challenging problem since video may be seriously distorted due to packet errors caused by wireless channels. The capability of predicting transmission distortion (i.e., video distortion caused by packet errors) can assist in designing video encoding and transmission schemes that achieve maximum video quality or minimum end-to-end video distortion. This paper is aimed at deriving formulas for predicting transmission distortion. The contribution of this paper is twofold. First, we identify the governing law that describes how the transmission distortion process evolves over time and analytically derive the transmission distortion formula as a closed-form function of video frame statistics, channel error statistics, and system parameters. Second, we identify, for the first time, two important properties of transmission distortion. The first property is that the clipping noise, which is produced by nonlinear clipping, causes decay of propagated error. The second property is that the correlation between motion-vector concealment error and propagated error is negative and has dominant impact on transmission distortion, compared with other correlations. Due to these two properties and elegant error/distortion decomposition, our formula provides not only more accurate prediction but also lower complexity than the existing methods.


Nonlinear endomicroscopy using a double-clad fiber coupler.
A double-clad fiber coupler is developed to be used in two-photon-excited fluorescence endomicroscopy to replace a dichroic mirror and separate the fluorescence signal from the excitation laser beam. With the double-clad fiber coupler, the endomicroscope becomes more compact, easier to be aligned, and more stable in alignment. The double-clad fiber coupler can transmit 62% of the excitation laser beam through the core. The fluorescence collection efficiency of the double-clad fiber coupler is 34%, which is, to the best of our knowledge, the highest fluorescence collection efficiency achieved by couplers used in two-photon-excited fluorescence endomicroscopes. As a result, the contrast of endomicroscopy imaging is enhanced.


Localization of breast cancer susceptibility loci by genome-wide SNP linkage disequilibrium mapping.
We studied the feasibility of a novel approach to localize breast cancer susceptibility genes, using a low-density genome-wide panel of single-nucleotide polymorphisms and taking advantage of large regions of linkage disequilibrium (LD) flanking Jewish disease genes in high-risk cases. With Affymetrix GeneChip arrays, we genotyped 8,576 polymorphisms in three sets of Ashkenazi Jewish breast cancer cases: a "validation" set of 27 breast cancer cases, all of whom carried the BRCA2*6174delT founder mutation; a "field" set of 19 breast cancer cases from male breast cancer kindreds, which simulated conditions for finding new genes; and a "test" set of 57 probands from breast cancer kindreds (4 or more cases/kindred), in which mutations in BRCA1 and BRCA2 had been excluded. To identify associations, we compared the frequency of genotypes and haplotypes in cases vs. controls by the Fisher's exact test and a maximum likelihood ratio test. In the "validation" set, we demonstrated the presence of a region of linkage disequilibrium on BRCA2*6174delT chromosomes that spanned over 5 million bases. In the "field" set, we showed that this large region of linkage disequilibrium flanking BRCA2 was detectable despite the presence of heterogeneity in the sample set. Finally, in the "test" set, at least three regions of interest emerged that could contain novel breast cancer genes, one of which had been identified previously by linkage analysis. While these results demonstrate the feasibility of genome-wide association strategies, further application of this approach will critically depend on optimizing the density and distribution of SNPs and the size and type of study design.


Ocean dynamics and acoustic fluctuations in the fram strait marginal ice zone.
Acoustic waves transmitted over a 100-kilometer path in the Fram Strait marginal ice zone undergo Doppler shifts and fluctuations around these shifts, the former due to quasi-steady motion of both acoustic source and receiver and the latter to unsteady motions of the water column and ice cover. Internal waves and differential Doppler shift usually account for such fluctuations in the deep temperate ocean but only partially explain the results obtained in the marginal ice zone. There the fluctuations are more energetic and may be caused alternatively or additionally by comparably energetic fluctuations in ice-edge eddies or other mesoscale motions.


Impact of increasing the relative availability of meat-free options on food selection: two natural field experiments and an online randomised trial.
Increasing the availability of lower energy-density foods is a promising intervention to encourage healthier food purchasing but few studies have examined the effect of increasing availability of meat-free meals to promote more sustainable purchasing. We report three studies, all examining the impact of altering the availability of meat-free meals on meal selection. Study 1 (a natural experiment in one university cafeteria) examined the impact of altering the ratio of meat-free meals (one meat-free and two meat, to two meat-free and one meat) on weekly sales of meals containing meat. Study 2 (a natural experiment in 18 worksite cafeterias) examined the impact on meat-free meal sales of a menu change designed to increase the availability of meat-free meals. Study 3 (an online study of 2205 UK-representative adults) compared meal selections when participants were randomised to ranges comprised of (a) one meat-free, three meat options; (b) two meat-free, two meat; or (c) three meat-free, one meat. Study 1 suggested a significant decrease in the proportion of sales of meat options when the availability of meat-free options increased (- 19.9 percentage points; 95%CIs:-25.2,-14.6), with no evidence of changes to meat-based meal sales in other university cafeterias during the same period. Findings from Study 2 were mixed: multilevel regressions found no evidence of an increase in meat-free meals following the menu change (2.3 percentage points; 95%CIs: - 1.3,5.9), while interrupted time-series analyses suggested sales did increase (2.3; 95%CIs: 0.4,4.2), but implementation of the planned change was limited. In Study 3 reducing meat-free options from 50 to 25% reduced participants' selection of meat-free options (odds ratio 0.35; 95%CIs: 0.26,0.46), while increasing meat-free options from 50 to 75% increased meat-free selections (odds ratio 2.43; 95%CIs: 1.94,3.04). There was no evidence effects were moderated by gender, socioeconomic status or usual meat consumption. Increasing the availability of meat-free options is effective at reducing meat selection and purchasing for different ratios of meat to meat-free options. The magnitude of the effect is uncertain, but with no evidence of differences in response by demographic groups when directly tested. Study 3: Open Science Framework; https://osf.io/ze9c6 ; 6/8/2020.


Comparative population dynamics of mosquito-borne flaviviruses.
Among the members of the genus Flavivirus are several important human pathogens, including the dengue (DEN) and Japanese encephalitis (JE) viruses. From the analysis of gene sequence data of samples of these virus populations it is possible to infer phylogenetic relationships, which in turn can yield important epidemiological information, including their demographic history in humans. In this study, we use a recently developed method, based on coalescent theory, to infer the population dynamics of a variety of mosquito-borne flaviviruses. Our study involves the testing of alternative hypotheses, the estimation of confidence intervals around demographic model parameter values, and the placing of the maximum likelihood (ML) demographic model into a "real time" epidemiological history. We reveal that all the Flavivirus populations studied are growing at an exponential rate, with the rates of population growth of dengue virus serotypes 2 and 3 increasing rapidly in the recent past, and that of Japanese encephalitis virus changing from constant population size to exponential growth within the last century. We therefore demonstrate that the use of these coalescent methods may be extremely valuable in monitoring responses to interventions such as vaccination or vector control.


LINKS: Scalable, alignment-free scaffolding of draft genomes with long reads.
Owing to the complexity of the assembly problem, we do not yet have complete genome sequences. The difficulty in assembling reads into finished genomes is exacerbated by sequence repeats and the inability of short reads to capture sufficient genomic information to resolve those problematic regions. In this regard, established and emerging long read technologies show great promise, but their current associated higher error rates typically require computational base correction and/or additional bioinformatics pre-processing before they can be of value. We present LINKS, the Long Interval Nucleotide K-mer Scaffolder algorithm, a method that makes use of the sequence properties of nanopore sequence data and other error-containing sequence data, to scaffold high-quality genome assemblies, without the need for read alignment or base correction. Here, we show how the contiguity of an ABySS Escherichia coli K-12 genome assembly can be increased greater than five-fold by the use of beta-released Oxford Nanopore Technologies Ltd. long reads and how LINKS leverages long-range information in Saccharomyces cerevisiae W303 nanopore reads to yield assemblies whose resulting contiguity and correctness are on par with or better than that of competing applications. We also present the re-scaffolding of the colossal white spruce (Picea glauca) draft assembly (PG29, 20 Gbp) and demonstrate how LINKS scales to larger genomes. This study highlights the present utility of nanopore reads for genome scaffolding in spite of their current limitations, which are expected to diminish as the nanopore sequencing technology advances. We expect LINKS to have broad utility in harnessing the potential of long reads in connecting high-quality sequences of small and large genome assembly drafts.


Linking ATM Promoter Methylation to Cell Cycle Protein Expression in Brain Tumor Patients: Cellular Molecular Triangle Correlation in ATM Territory.
Ataxia telangiectasia mutated (ATM) is a key gene in DNA double-strand break (DSB), and therefore, most of its disabling genetic alterations play an important initiative role in many types of cancer. However, the exact role of ATM gene and its epigenetic alterations, especially promoter methylation in different grades of brain tumors, remains elusive. The current study was conducted to query possible correlations among methylation statue of ATM gene, ATM/ retinoblastoma (RB) protein expression, D1853N ATM polymorphism, telomere length (TL), and clinicopathological characteristics of various types of brain tumors. Isolated DNA from 30 fresh tissues was extracted from different types of brain tumors and two brain tissues from deceased normal healthy individuals. DNAs were treated with bisulfate sodium using DNA modification kit (Qiagen). Methylation-specific polymerase chain reaction (MSP-PCR) was implicated to determine the methylation status of treated DNA templates confirmed by promoter sequencing. Besides, the ATM and RB protein levels were determined by immunofluorescence (IF) assay using monoclonal mouse antihuman against ATM, P53, and RB proteins. To achieve an interactive correlation, the methylation data were statistically analyzed by considering TL and D1853N ATM polymorphism. More than 73% of the brain tumors were methylated in ATM gene promoter. There was strong correlation between ATM promoter methylation and its protein expression (p < 0.001). As a triangle, meaningful correlation was also found between methylated ATM promoter and ATM protein expression with D1853N ATM polymorphism (p = 0.01). ATM protein expression was not in line with RB protein expression while it was found to be significantly correlated with ATM promoter methylation (p = 0.01). There was significant correlation between TL neither with ATM promoter methylation nor with ATM protein expression nor with D1853N polymorphism. However, TL has shown strong correlation with patient's age and tumor grade (p = 0.01). Given the important role of cell cycle checkpoint proteins as well as RB and ATM in TL and cancer evolution, further assessment is warranted to shed more light on the pathway linking the telomere instability to tumor progression. High ATM methylation rate in brain tumor patients could open a new avenue toward early screening and cancer therapy.


Combined effect of posttraumatic stress disorder and prescription opioid use on risk of cardiovascular disease.
Prescription opioid analgesic use (OAU) is associated with increased risk of cardiovascular disease (CVD). OAU is more common in patients with than without posttraumatic stress disorder (PTSD), and PTSD is associated with higher CVD risk. We determined whether PTSD and OAU have an additive or multiplicative association with incident CVD. Veterans Health Affairs patient medical record data from 2008 to 2015 was used to identify 2861 patients 30-70 years of age, free of cancer, CVD and OAU for 12 months before index date. We defined a four-level exposure variable: 1) no PTSD/no OAU, 2) OAU alone, 3) PTSD alone and 4) PTSD+OAU. Cox proportional hazard models estimated the association between the exposure variable and incident CVD. The mean age was 49.0 (±11.0), 85.7% were male and 58.3% were White, 34.4% had no PTSD/no OAU, 32.9% had PTSD alone, 10.6% had OAU alone, and 22.1% had PTSD+OAU. Compared with patients with no PTSD/no OAU, those with PTSD alone were not at increased risk of incident CVD (hazard ratio = 0.82; 95% confidence interval (CI): 0.63-1.17); however, OAU alone and PTSD+OAU were both significantly associated with incident CVD (hazard ratio = 1.99; 95% CI:1.36-2.92 and hazard ratio = 2.20; 95% CI: 1.61-3.02). There was no significant additive or multiplicative PTSD and OAU association with incident CVD. OAU is associated with nearly a two-fold increased risk of CVD in patients with and without PTSD. Despite no additive or multiplicative interaction effects, the high prevalence of OAU in PTSD may represent a novel contributor to the elevated CVD burden among patients with PTSD.


Purification of HCC-specific extracellular vesicles on nanosubstrates for early HCC detection by digital scoring.
We report a covalent chemistry-based hepatocellular carcinoma (HCC)-specific extracellular vesicle (EV) purification system for early detection of HCC by performing digital scoring on the purified EVs. Earlier detection of HCC creates more opportunities for curative therapeutic interventions. EVs are present in circulation at relatively early stages of disease, providing potential opportunities for HCC early detection. We develop an HCC EV purification system (i.e., EV Click Chips) by synergistically integrating covalent chemistry-mediated EV capture/release, multimarker antibody cocktails, nanostructured substrates, and microfluidic chaotic mixers. We then explore the translational potential of EV Click Chips using 158 plasma samples of HCC patients and control cohorts. The purified HCC EVs are subjected to reverse-transcription droplet digital PCR for quantification of 10 HCC-specific mRNA markers and computation of digital scoring. The HCC EV-derived molecular signatures exhibit great potential for noninvasive early detection of HCC from at-risk cirrhotic patients with an area under receiver operator characteristic curve of 0.93 (95% CI, 0.86 to 1.00; sensitivity = 94.4%, specificity = 88.5%).


Bias in the reporting of harms in clinical trials of second-generation antidepressants for depression and anxiety: A meta-analysis.
Previous research has shown that reporting bias has inflated the apparent efficacy of antidepressants. We investigated whether apparent safety was also affected. We included 133 trials, involving 31,296 patients, of second-generation antidepressants for the treatment of major depressive disorder (MDD) or anxiety disorders, obtained from Food and Drug Administration (FDA) reviews. We extracted data on overall discontinuation, discontinuation due to adverse events, and serious adverse events (SAEs). Meta-analysis was used to compare discontinuation rates between FDA reviews and matching journal articles, while SAEs were compared qualitatively. The odds ratio for overall discontinuation, comparing drug to placebo, was 1.0 for both sources, while that for discontinuation due to adverse events was 2.4 for both sources. Seventy-seven of 97 (79%) journal articles provided incomplete information on SAEs; sixty-one (63%) articles made no mention of SAEs at all. Of 21 articles which could be compared to the FDA, only 6 (29%) had full reporting without discrepancies. Nine (43%) articles reported a discrepant number of SAEs. Descriptions were absent or discrepant in 6 (29%) additional articles, even for important SAEs such as suicide attempts. In conclusion, reporting bias has not affected average discontinuation rates over trials. However, SAE reporting is not only very poor, with over half of articles failing to discuss SAEs altogether, but discrepancies between the FDA and articles were common and often led to a more favorable drug-placebo comparison. These findings suggest that journal articles are an unreliable source of data on SAEs in antidepressant trials.


Resistance to Schistosoma mansoni in humans: influence of the IgE/IgG4 balance and IgG2 in immunity to reinfection after chemotherapy.
The hypothesis of an association between human resistance to reinfection by the parasite Schistosoma mansoni and anti-larval immunoglobulin isotypes was tested by logistic regression in the presence of the explicative variables water contact, age, and sex. Of the seven isotypes tested (IgM, IgG1, IgG2, IgG3, IgG4, IgA, and IgE), only IgE, IgG4, and IgG2 showed an association (positive for IgE and negative for IgG2 and IgG4) with resistance to reinfection after chemotherapy. The opposite effects of IgE and IgG4 were undissociable in the analysis, indicating that these isotypes probably antagonize each other in protection. The negative association of IgG2 with resistance is consistent with the view that anti-carbohydrate antibodies may facilitate reinfection. Finally, epidemiologic and immunologic studies support the view that there is a progressive but slow development of acquired immunity in children and adolescents.


COURTSHIP SOUND PRODUCTION IN TWO SYMPATRIC SIBLING DROSOPHILA SPECIES.
Courting males of Drosophila persimilis produce a pulsed "vibration sound," with frequency of oscillation and pulse repetition rate markedly different from that of D. pseudoobscura. These differences probably contribute to reproductive isolation. Flight frequency is the same in both species. Brief "preliminary sounds" are common and the same in both species. Males with large parts of both wings removed produce nearly normal vibration sounds, and normal preliminary sounds.


Tomato annexins p34 and p35 bind to F-actin and display nucleotide phosphodiesterase activity inhibited by phospholipid binding.
Annexins are a family of proteins found in a range of eukaryotic cell types. They share a characteristic amino acid sequence and a Ca(2+)-dependent affinity for specific phospholipids. In plants, proteins with common properties and significant homology with annexins have been identified in a number of species and implicated in diverse cellular functions known to be modulated by Ca2+. This study describes several novel biochemical properties of the tomato annexins p34 and p35 that are relevant to our understanding of their functions in the plant. First, the annexins were found to bind to actin in a calcium- and pH-dependent interaction that was specific for F-actin and not G-actin. Second, an enzyme activity defined as a nucleotide phosphodiesterase activity was found associated with the purified annexin preparation. Selective immunoprecipitation of p34 and p35 strongly suggests that the enzyme activity is a property of the annexins and constitutes 60% of the total soluble activity found in root extracts capable of hydrolyzing free ATP. The substrate specificity of the enzyme within in vitro assays is broad. ATP is the preferred substrate, but nearly identical rates of hydrolysis of GTP and substantial hydrolysis of other nucleotide tri- and diphosphates are observed. The enzyme activity was found to be a property of both p34 and p35, although the specific activity was routinely higher for p34. Third, the enzyme activity of the annexins was not affected by F-actin binding but could be abolished by the specific Ca(2+)-dependent interaction of the annexins with phospholipids. Our results showed that p34 and p35 account for substantial enzyme activity in tomato root cells. This activity was exhibited when the proteins were either in soluble form or attached to actin filaments. Enzyme activity was not exhibited when the annexins were bound to phospholipids. These properties suggest a role for the proteins in mediating Ca(2+)-dependent events involving interactions of the cytoskeleton and cellular membranes.


Gene reactivation: a tool for the isolation of mammalian DNA methylation mutants.
We report the isolation and characterization of a mammalian strain (tsm) that has a temperature-sensitive mutation in DNA methylation. The isolation procedure was based on the observation that treatment of a CHO TK- MT- cell line with demethylating agents introduces up to 46% demethylation, resulting in phenotypic reversion and transcriptional activation of the thymidine kinase (TK) and metallothionein (MT) genes at frequencies ranging from 1% to 59%. Seven thousand individual colonies from an EMS-mutagenized CHO TK- MT- population were screened for spontaneous reversion to TK+ phenotype after treatment at 39 degrees C. Successful isolates were subsequently examined for MT+ reversion. A single clone (tsm) was obtained that showed temperature-dependent reactivation of both TK and MT genes at frequencies of 7.2 X 10(-4) and 6 X 10(-4), respectively. The tsm cells were viable at 39 degrees C and showed no increased mutation frequency. Reactivation correlated with transcriptional activation of the respective genes, whereas backreversion to the TK- phenotype was associated with transcriptional inactivation. TK- backrevertants were reactivable again with demethylating agents. Although demethylation in tsm cells was not detectable by HPLC, Southern blot analysis revealed that reactivants, irrespective of their mode of generation, showed specific demethylation of both TK and MT genes. Also, after about 150 cell generations after treatment, reactivants from both temperature-induced tsm and cells exposed to demethylating agents gained 60% and 23%, respectively, in 5-methylcytosine (5mC). It is proposed that the phenotype of tsm cells is due to a mutation involved in the regulation of DNA methylation. The further characterization of this and other mammalian mutants should help to clarify the physiological role of DNA methylation, as well as its regulation.


Effect of ingestion of fat on ileostomy effluent.
Infusion of fat into the ileum slows small bowel transit and increases absorption of a carbohydrate meal. Paired studies were undertaken to investigate the effect of adding fat to the diet on gastrointestinal transit and absorption in eight people with terminal ileostomies. Each subject ate a daily diet that contained either low or high levels of fat but equivalent amounts of carbohydrate, protein, and fibre. Solid and liquid markers were added to the food to measure transit times. The ileostomy outputs after the meals were analysed for protein, fat, carbohydrate, polyethylene glycol, and the delivery of solid markers. All subjects produced more ileal effluent while taking the high fat diet compared with the low fat diet. Fat and protein outputs were significantly higher on the high fat days, but a greater proportion of the fat was absorbed during the high fat diet compared with the low fat diet. The output of carbohydrate on the two diets was not significantly different. The incorporation of fat in the diet produced no significant differences in the transit times of the first marker, but a slight prolongation of the transit time of the second marker. Increasing the fat content of the diet did not slow small bowel transit or increase nutrient absorption. The increase in protein and water output could be explained by an increased secretion of pancreatic enzymes.


Clinical prediction rules in practice: review of clinical guidelines and survey of GPs.
The publication of clinical prediction rules (CPRs) studies has risen significantly. It is unclear if this reflects increasing usage of these tools in clinical practice or how this may vary across clinical areas. To review clinical guidelines in selected areas and survey GPs in order to explore CPR usefulness in the opinion of experts and use at the point of care. A review of clinical guidelines and survey of UK GPs. Clinical guidelines in eight clinical domains with published CPRs were reviewed for recommendations to use CPRs including primary prevention of cardiovascular disease, transient ischaemic attack (TIA) and stroke, diabetes mellitus, fracture risk assessment in osteoporosis, lower limb fractures, breast cancer, depression, and acute infections in childhood. An online survey of 401 UK GPs was also conducted. Guideline review: Of 7637 records screened by title and/or abstract, 243 clinical guidelines met inclusion criteria. CPRs were most commonly recommended in guidelines regarding primary prevention of cardiovascular disease (67%) and depression (67%). There was little consensus across various clinical guidelines as to which CPR to use preferentially. Of 401 responders to the GP survey, most were aware of and applied named CPRs in the clinical areas of cardiovascular disease and depression. The commonest reasons for using CPRs were to guide management and conform to local policy requirements. GPs use CPRs to guide management but also to comply with local policy requirements. Future research could focus on which clinical areas clinicians would most benefit from CPRs and promoting the use of robust, externally validated CPRs.


Agonist-induced redistribution of bradykinin B2 receptor in caveolae.
Redistribution of receptors within the plasma membrane as well as between the plasma membrane and various cell compartments presents an important way of regulating the cellular responsiveness to their cognate agonists. We have applied immunocytochemical methods to localize the bradykinin B2 receptor and to examine its agonist induced redistribution in A431 cells. In situ labeling with antibodies to ectodomain-2 of the receptor which do not interfere with bradykinin binding of the receptor showed a random distribution of the B2 receptor on the plasma membrane. Stimulation of cells with 20 nM bradykinin markedly reduced the accessibility of the antibody to its corresponding epitope in non-permeabilized cells. Immuno-electron microscopy revealed the presence of receptors in membrane-near vesicles that are surrounded by an electron-transparent halo. Fluorescence microscopic double labeling co-localized the B2 receptor protein with caveolin-1 by a convergent pattern of punctate staining. At the ultrastructural level the B2 receptor protein was found in vesicles that bear the immunolabel of caveolin-1 and display the morphological characteristics of caveolae. We conclude that stimulation of B2 receptors results in their redistribution and sequestration in caveolae, an event that is likely to be implicated in receptor signaling and/or desensitization. The localization of B2 receptors in endosome-like structures after prolonged exposure to bradykinin might indicate that the internalization through caveolae may communicate with other endocytotic pathways of A431 cells.


Clinical accuracy and initial experience with augmented reality-assisted pedicle screw placement: the first 205 screws.
Augmented reality (AR) is a novel technology which, when applied to spine surgery, offers the potential for efficient, safe, and accurate placement of spinal instrumentation. The authors report the accuracy of the first 205 pedicle screws consecutively placed at their institution by using AR assistance with a unique head-mounted display (HMD) navigation system. A retrospective review was performed of the first 28 consecutive patients who underwent AR-assisted pedicle screw placement in the thoracic, lumbar, and/or sacral spine at the authors' institution. Clinical accuracy for each pedicle screw was graded using the Gertzbein-Robbins scale by an independent neuroradiologist working in a blinded fashion. Twenty-eight consecutive patients underwent thoracic, lumbar, or sacral pedicle screw placement with AR assistance. The median age at the time of surgery was 62.5 (IQR 13.8) years and the median body mass index was 31 (IQR 8.6) kg/m2. Indications for surgery included degenerative disease (n = 12, 43%); deformity correction (n = 12, 43%); tumor (n = 3, 11%); and trauma (n = 1, 4%). The majority of patients (n = 26, 93%) presented with low-back pain, 19 (68%) patients presented with radicular leg pain, and 10 (36%) patients had documented lower extremity weakness. A total of 205 screws were consecutively placed, with 112 (55%) placed in the lumbar spine, 67 (33%) in the thoracic spine, and 26 (13%) at S1. Screw placement accuracy was 98.5% for thoracic screws, 97.8% for lumbar/S1 screws, and 98.0% overall. AR depicted through a unique HMD is a novel and clinically accurate technology for the navigated insertion of pedicle screws. The authors describe the first 205 AR-assisted thoracic, lumbar, and sacral pedicle screws consecutively placed at their institution with an accuracy of 98.0% as determined by a Gertzbein-Robbins grade of A or B.


Photolabile protecting groups in metal-organic frameworks: preventing interpenetration and masking functional groups.
Photolabile groups can be incorporated into metal-organic frameworks (MOFs) and then quantitatively cleaved following MOF formation. Here, a 2-nitrobenzyl ether prevents lattice interpenetration (catenation) in a cubic MOF derived from zinc(II). Subsequent photolysis unmasks a hydroxyl group, and produces an open MOF that cannot be synthesized directly.


Consumption of molecular hydrogen prevents the stress-induced impairments in hippocampus-dependent learning tasks during chronic physical restraint in mice.
We have reported that hydrogen (H(2)) acts as an efficient antioxidant by gaseous rapid diffusion. When water saturated with hydrogen (hydrogen water) was placed into the stomach of a rat, hydrogen was detected at several microM level in blood. Because hydrogen gas is unsuitable for continuous consumption, we investigated using mice whether drinking hydrogen water ad libitum, instead of inhaling hydrogen gas, prevents cognitive impairment by reducing oxidative stress. Chronic physical restraint stress to mice enhanced levels of oxidative stress markers, malondialdehyde and 4-hydroxy-2-nonenal, in the brain, and impaired learning and memory, as judged by three different methods: passive avoidance learning, object recognition task, and the Morris water maze. Consumption of hydrogen water ad libitum throughout the whole period suppressed the increase in the oxidative stress markers and prevented cognitive impairment, as judged by all three methods, whereas hydrogen water did not improve cognitive ability when no stress was provided. Neural proliferation in the dentate gyrus of the hippocampus was suppressed by restraint stress, as observed by 5-bromo-2'-deoxyuridine incorporation and Ki-67 immunostaining, proliferation markers. The consumption of hydrogen water ameliorated the reduced proliferation although the mechanistic link between the hydrogen-dependent changes in neurogenesis and cognitive impairments remains unclear. Thus, continuous consumption of hydrogen water reduces oxidative stress in the brain, and prevents the stress-induced decline in learning and memory caused by chronic physical restraint. Hydrogen water may be applicable for preventive use in cognitive or other neuronal disorders.


Psychometric properties and performance of the 17-item Benefit Finding Scale (BFS) in an outpatient population of men with prostate cancer.
To analyse the psychometric properties and performance of the 17-item Benefit Finding Scale (BFS) in an Australian outpatient sample of men with prostate cancer. The instrument's psychometric properties and performance were rated against established criteria for reliability (internal consistency), construct validity (instrument dimensionality) and variability (floor and ceiling effects). Internal consistency reliability was satisfactory as evidenced by a Cronbach's alpha of 0.95 Dimensionality analysis confirmed a unidimensional structure indicating construct validity. A greater than 15% floor effect suggested limited data variability. The 17-item BFS seems to have satisfactory psychometric properties for use in an outpatient sample of men with prostate cancer, with some questions regarding detection of variability. The high internal consistency reliability points to the instruments ability to reliably capture the benefit finding construct in this population. The evidence for instrument dimensionality indicates a unidimensional scale, and thus a calculation of a single total score can be recommended. The >15% floor effect suggests that there may be issues with the instrument's ability to detect variance, and thus some questions remain regarding the instrument's ability to discern change in health status over time. Nevertheless, the findings of this study together with previous evidence indicate that the 17-item BFS can be recommended as the tool of choice when exploring benefit finding in adult cancer populations.


Identification and characterization of the pseudopterosin diterpene cyclase, elisabethatriene synthase, from the marine gorgonian, Pseudopterogorgia elisabethae.
The pseudopterosins are diterpene glycosides isolated from the marine gorgonian, Pseudopterogorgia elisabethae, which exhibit anti-inflammatory and analgesic activity greater than the industry standard, indomethacin. Previously, we isolated the pseudopterosin diterpene cyclase product, elisabethatriene, using a radioactivity-guided isolation. Identification of this metabolite, and the conversion of labeled geranylgeranyl diphosphate to elisabethatriene, provided us with an assay to guide the isolation of the enzyme responsible for this cyclization. The soluble protein preparation from P. elisabethae has been partially purified (approximately 15,000-fold) using a combination of low-resolution anion-exchange, low-resolution hydrophobic interaction, high-resolution hydroxyapatite, and high-resolution anion-exchange chromatography. The diterpene cyclase was identified by comparing the molecular weight from gel permeation chromatography (approximately 47,000Da) with those of protein bands from purified fractions using SDS-PAGE gel electrophoresis. Kinetic analysis and evaluation of amino acid inhibition studies indicated that the enzyme displays similar characteristics to other terpenoid cyclases isolated from terrestrial sources. This report represents the first purification and characterization of a terpene biosynthetic enzyme from a marine invertebrate.


A well-resolved phylogeny of the trees of Puerto Rico based on DNA barcode sequence data.
The use of phylogenetic information in community ecology and conservation has grown in recent years. Two key issues for community phylogenetics studies, however, are (i) low terminal phylogenetic resolution and (ii) arbitrarily defined species pools. We used three DNA barcodes (plastid DNA regions rbcL, matK, and trnH-psbA) to infer a phylogeny for 527 native and naturalized trees of Puerto Rico, representing the vast majority of the entire tree flora of the island (89%). We used a maximum likelihood (ML) approach with and without a constraint tree that enforced monophyly of recognized plant orders. Based on 50% consensus trees, the ML analyses improved phylogenetic resolution relative to a comparable phylogeny generated with Phylomatic (proportion of internal nodes resolved: constrained ML = 74%, unconstrained ML = 68%, Phylomatic = 52%). We quantified the phylogenetic composition of 15 protected forests in Puerto Rico using the constrained ML and Phylomatic phylogenies. We found some evidence that tree communities in areas of high water stress were relatively phylogenetically clustered. Reducing the scale at which the species pool was defined (from island to soil types) changed some of our results depending on which phylogeny (ML vs. Phylomatic) was used. Overall, the increased terminal resolution provided by the ML phylogeny revealed additional patterns that were not observed with a less-resolved phylogeny. With the DNA barcode phylogeny presented here (based on an island-wide species pool), we show that a more fully resolved phylogeny increases power to detect nonrandom patterns of community composition in several Puerto Rican tree communities. Especially if combined with additional information on species functional traits and geographic distributions, this phylogeny will (i) facilitate stronger inferences about the role of historical processes in governing the assembly and composition of Puerto Rican forests, (ii) provide insight into Caribbean biogeography, and (iii) aid in incorporating evolutionary history into conservation planning.


Chromosomal-level assembly of yellow catfish genome using third-generation DNA sequencing and Hi-C analysis.
The yellow catfish, Pelteobagrus fulvidraco, belonging to the Siluriformes order, is an economically important freshwater aquaculture fish species in Asia, especially in Southern China. The aquaculture industry has recently been facing tremendous challenges in germplasm degeneration and poor disease resistance. As the yellow catfish exhibits notable sex dimorphism in growth, with adult males about two- to three-fold bigger than females, the way in which the aquaculture industry takes advantage of such sex dimorphism is another challenge. To address these issues, a high-quality reference genome of the yellow catfish would be a very useful resource. To construct a high-quality reference genome for the yellow catfish, we generated 51.2 Gb short reads and 38.9 Gb long reads using Illumina and Pacific Biosciences (PacBio) sequencing platforms, respectively. The sequencing data were assembled into a 732.8 Mb genome assembly with a contig N50 length of 1.1 Mb. Additionally, we applied Hi-C technology to identify contacts among contigs, which were then used to assemble contigs into scaffolds, resulting in a genome assembly with 26 chromosomes and a scaffold N50 length of 25.8 Mb. Using 24,552 protein-coding genes annotated in the yellow catfish genome, the phylogenetic relationships of the yellow catfish with other teleosts showed that yellow catfish separated from the common ancestor of channel catfish ∼81.9 million years ago. We identified 1,717 gene families to be expanded in the yellow catfish, and those gene families are mainly enriched in the immune system, signal transduction, glycosphingolipid biosynthesis, and fatty acid biosynthesis. Taking advantage of Illumina, PacBio, and Hi-C technologies, we constructed the first high-quality chromosome-level genome assembly for the yellow catfish P. fulvidraco. The genomic resources generated in this work not only offer a valuable reference genome for functional genomics studies of yellow catfish to decipher the economic traits and sex determination but also provide important chromosome information for genome comparisons in the wider evolutionary research community.


Enhancing survival outcomes in the management of patients with higher-risk myelodysplastic syndromes.
The myelodysplastic syndromes (MDS) are a collection of clonal myeloid neoplasms characterized by bone marrow failure and cytopenias. Patients classified with higher-risk disease include those with intermediate-2 or high-risk disease as classified by the International Prognostic Scoring System (IPSS). These patients represent 29% of the MDS population and were originally reported to have a median survival without therapy of only 0.4 for high-risk patients and 1.1 years for intermediate-2 risk patients. The most significant treatment goals in these patients involve prolonging the time to acute myeloid leukemia progression and extending overall survival. Quality of life, symptom control, and transfusion independence are also important. The National Comprehensive Cancer Network guidelines divide treatment options for this population into high- and low-intensity therapies. High-intensity therapies include cytarabine-based remission induction chemotherapy and hematopoietic stem cell transplantation. Transplantation is the only treatment option with the ability to cure; however, many MDS patients may not qualify due to age or comorbidities. Low-intensity therapies include the DNA methyltransferase inhibitors azacitidine and decitabine. Recently, azacitidine demonstrated the ability to extend survival by as much as 74% despite a modest complete response rate. This review examines the classification and diagnosis of higher-risk MDS patients, the management goals for these patients, clinical experience involved with treatments, guidelines, and recommendations for therapeutic options.


Plants know where it hurts: root and shoot jasmonic acid induction elicit differential responses in Brassica oleracea.
Plants respond to herbivore attack by rapidly inducing defenses that are mainly regulated by jasmonic acid (JA). Due to the systemic nature of induced defenses, attack by root herbivores can also result in a shoot response and vice versa, causing interactions between above- and belowground herbivores. However, little is known about the molecular mechanisms underlying these interactions. We investigated whether plants respond differently when roots or shoots are induced. We mimicked herbivore attack by applying JA to the roots or shoots of Brassica oleracea and analyzed molecular and chemical responses in both organs. In shoots, an immediate and massive change in primary and secondary metabolism was observed. In roots, the JA-induced response was less extensive and qualitatively different from that in the shoots. Strikingly, in both roots and shoots we also observed differential responses in primary metabolism, development as well as defense specific traits depending on whether the JA induction had been below- or aboveground. We conclude that the JA response is not only tissue-specific but also dependent on the organ that was induced. Already very early in the JA signaling pathway the differential response was observed. This indicates that both organs have a different JA signaling cascade, and that the signal eliciting systemic responses contains information about the site of induction, thus providing plants with a mechanism to tailor their responses specifically to the organ that is damaged.


Identification of Patient Characteristics Influencing Setting of Care Decisions for Patients With Acute Bacterial Skin and Skin Structure Infections: Results of a Discrete Choice Experiment.
Rates of acute bacterial skin and skin structure infections (ABSSSI) have sharply increased since 2000. Treatment may be administered in the inpatient or outpatient setting; clinical decision-making regarding hospitalization is inconsistent, often leading to hospitalization of some patients with ABSSSI who qualify for outpatient parenteral antimicrobial therapy, which leads to increased overall care costs. New antibiotics such as oritavancin are hypothesized to be a cost-effective option improving accessibility to ambulatory treatment of ABSSSI. The goal of this study was to understand the patient attributes that affect clinical decision-making regarding the setting of care for ABSSSI treatment. An observational, cross-sectional study was conducted that surveyed clinicians of various specialties from the United States and the United Kingdom. The survey collected quantitative responses and used a series of choice-based experimental designs to evaluate patient attributes influencing clinical treatment decisions. Infection severity, severe comorbidities, and age ≥ 75 years were observed to have the greatest impact on treatment location decisions (odds ratio [OR], 0.000-0.004 [95% CI, 0.000-0.011], vs mild ABSSSI; OR, 0.246-0.484 [95% CI, 0.154-0.788], vs no active comorbidities; OR, 0.136-0.523 [95% CI, 0.070-0.888], vs ≤ 18 years, respectively). The majority of respondents indicated they would consider oritavancin to avoid postdischarge outpatient parenteral antimicrobial therapy or oral therapy, regardless of the pathogen (63.5%-83.5%). Key factors influencing ABSSSI treatment setting were severity of infection, severity of comorbidities, and age. Clinicians surveyed identified patient profiles in which single-dose oritavancin might enable wholly outpatient or shortened inpatient management. Additional studies to elucidate the ABSSSI care pathways that include oritavancin and other novel antibiotics are needed.


The effects of Anethum graveolens essence on scopolamine-induced memory impairment in mice.
Since Anethum graveolens (Dill) has phytoestrogenic compounds and it is proven that estrogens exert beneficial effects on cognition; the aim of this study was to understand if this plant can improve memory performance. Male Balb/c mice weighing 25-30 g were used in this study and memory was assessed by the novel object recognition task. In this method, the difference in the exploration time between a familiar object and a novel object is taken as an index of memory performance (recognition index, RI). Scopolamine significantly reduced memory index (RI = -15.5% ± 3.0). Dill essence (100 mg/kg, ip) prevented the harmful effects of scopolamine on memory (RI = 40% ± 5.5), thus RI did not differ with control animals (RI = 50% ± 5.8). In addition, 17-β estradiol also prevented memory impairment in animals (0.2 mg/kg, ip; RI = 35.8% ± 6.5). Nevertheless, the beneficial effects of dill essence were antagonized by prior injection of tamoxifen (1 mg/kg, ip; RI = -30% ± 7.8). Although phytoesrogens are not steroids, the beneficial effect of dill on memory, at least in part, may have been achieved by estrogenic receptors present in the brain. Thus dill essence could be promising in improving memory and cognition, mainly in postmenopausal women.


Aquaporin-4 in hepatic encephalopathy.
Brain edema is a critical component of hepatic encephalopathy (HE) associated with acute liver failure and such edema appears to be principally due to astrocyte swelling (cytotoxic edema). Ammonia is believed to represent a major factor responsible for astrocyte swelling, although the mechanisms by which ammonia causes such swelling are not completely understood. Recent studies have implicated potential role of oxidative stress, and the mitochondrial permeability transition (mPT). While it is not known how oxidative stress and the mPT cause astrocyte swelling, it is reasonable to suggest that these events may affect one or more plasma membrane proteins involved in water and ion homeostasis in astrocytes. One such protein strongly implicated in brain edema in other neurological conditions is the water channel protein aquaporin-4 (AQP-4), which is abundantly expressed in astrocytes. This article summarizes the potential role of AQP-4 in brain edema in in vivo models of HE, as well as in ammonia-induced cell swelling in cultured astrocytes. The involvement of AQP-4 in the effects of manganese, another toxin implicated in HE, will also be discussed.


Women with eating problems: a community survey.
A survey was conducted of the eating habits and attitudes of a sample of young adult women. Each woman was asked whether she considered herself to have an eating problem. The sample comprised 369 attenders at a family planning clinic. Self-report assessment procedures were used, including the Eating Attitudes Test (EAT) and the General Health Questionnaire (GHQ). Eating problems were reported by 20.6 per cent of the sample. Factors which strongly discriminated between those with an eating problem and the remainder were binge eating and a high score on the EAT. Other important factors were laxative use and high levels of psychological morbidity. Self-induced vomiting was only weakly associated with reporting an eating problem, and past and present weight were unrelated to such reports.


Reduced negative surface charge on arterial endothelium explains accelerated atherosclerosis in type 2 diabetic patients.
Reduced endothelial surface charge markedly increases the rate of LDL uptake into blood vessels. Previous work in the streptozotocin diabetic rat reported reduced endothelial surface charge. We compared endothelial surface charge density in internal mammary artery rings from patients with type 2 diabetes (n = 12) and from non diabetic patients undergoing coronary artery bypass grafting, and observed a substantial (52%) reduction in the former. This was associated with higher plasma sialic acid levels suggesting loss of sialic acid residues from the glycocalyx as a possible mechanism.


New implications for the origin of the IAB main group iron meteorites and the isotopic evolution of the noncarbonaceous (NC) reservoir.
The origin of the IAB main group (MG) iron meteorites is explored through consideration of 182W isotopic compositions, thermal modeling of 26Al decay, and mass independent (nucleosynthetic) Mo isotopic compositions of planetesimals formed in the noncarbonaceous (NC) protosolar isotopic reservoir. A refined 182W model age for the meteorites Campo del Cielo, Canyon Diablo, and Nantan suggests that the IAB-MG parent body underwent some form of metal-silicate segregation as early as 5.3 ± 0.4 Myr after calcium-aluminum rich inclusion (CAI) formation or as late as 13.8 ± 1.4 Myr after CAI formation. If melting of the IAB-MG occurred prior to 7 Myr after CAI formation, it was likely driven by 26Al decay for a parent body radius >40 km. Otherwise, additional heat from impact is required for melting metal this late in Solar System history. If melting was partially or wholly the result of internal heating, a thermal model of 26Al decay heat production constrains the accretion age of the IAB-MG parent body to ~1.7 ± 0.4 Myr after CAI formation. If melting was, instead, dominantly caused by impact heating, thermal modeling suggests the parent body accreted more than 2 Myr after CAI formation. Comparison of Mo mass independent isotopic compositions of the IAB-MG to other NC bodies with constrained accretion ages suggests that the Mo isotopic composition of the NC reservoir changed with time, and that the IAB-MG parent body accreted between 2 to 3 Myr after CAI formation, thus requiring an origin by impact. The relationship between nucleosynthetic Mo isotopic compositions and accretion ages of planetesimals from the NC reservoir suggests that isotopic heterogeneity developed from either addition of s-process material to, or removal of coupled r-/p-process material from the NC reservoir.


A systematic review of predictors, moderators, and mediators of augmentative and alternative communication (AAC) outcomes for children with autism spectrum disorder.
Documented variability in augmentative and alternative communication (AAC) intervention outcomes makes it difficult for clinicians to select systems most likely to be effective for individual children with autism spectrum disorder (ASD). The aim of this study was to identify child-related factors associated with AAC intervention outcomes through a systematic review of the research literature. A search was conducted of peer-reviewed research articles in which AAC intervention outcomes and associated factors were reported for children with ASD. The search yielded 965 titles and abstracts, of which seven articles relating to six studies met criteria for inclusion. In total, 18 factors were examined, of which nine were assessed as predictors (e.g., cognition, ASD severity, language use), three as moderators (e.g., joint attention, object exploration), and six as mediators (e.g., frequency of therapy, communication partner knowledge). Child characteristics associated with communication outcomes were pre-intervention cognition, severity of ASD, verbal imitation, vocabulary comprehension, object use, joint attention, language use, and two multi-dimensional measures of communication competence. This study suggests emerging evidence for predictors, but less is known about what factors moderate and mediate response to AAC interventions.


Relationships and betrayal among young women: theoretical perspectives on adolescent dating abuse.
Adolescent dating abuse is not specifically described by any current nursing theory, and this article presents discussion of some existing theories that could inform a nursing theory of adolescent dating abuse. To account for the effects of gender, this discussion is limited to young women. Adolescent dating abuse is an important and understudied international issue for nursing. Theoretical frameworks can support development of nursing scholarship for such issues. No single theory yet exists in nursing to explain the experiences and health ramifications of dating abuse among young women. A summary table of theories is provided. Literature was gathered via database search and bibliographic snowballing from reference lists of relevant articles. Included literature dates from 1982 through 2010. Theories of relationship formation and function are discussed, including attachment, investment, feminist and gender role conflict theories. Betrayal trauma theory is considered as a mechanism of injury following an abusive dating experience. Gender, relationship and adolescence combine in a complex developmental moment for young women. To improve nursing care for those at risk for or in the throes of abusive relationships, it is critical to develop specific nursing approaches to understanding these relationships. Existing theories related to relationship and traumatic experiences can be combined in the development of a nursing theory of adolescent dating abuse among young women.


The Effect of Casein Protein Prior to Sleep on Fat Metabolism in Obese Men.
We have previously shown that ingesting protein at night before sleep is either beneficial or non-detrimental to metabolism, health, and body composition in obese women. However, the overnight protein-induced lipolytic actions and mechanism for improved metabolism and body composition have not been fully established. Therefore, in a crossover design, twelve obese men (age, 27.0 ± 2.2 years) were randomly assigned to ingest (within 30 min of sleep) casein protein (CAS, 120 kcal) or a non-nutritive placebo (PLA) before going to sleep. Markers of fat metabolism (lipolysis, substrate utilization, growth hormone), insulin, glucose, resting energy expenditure (REE), and appetite (questionnaire and ghrelin) were measured. During sleep and the next morning, interstitial glycerol from the subcutaneous abdominal adipose tissue (SCAAT) was measured using microdialysis. There were no differences in SCAAT glycerol (overnight: CAS, 177.4 ± 26.7; PLA, 183.8 ± 20.2 μmol/L; morning: CAS, 171.6 ± 19.1; PLA, 161.5 ± 18.6 μmol/L), substrate utilization, REE, or any blood markers between CAS and PLA. Desire to eat was greater for CAS compared to baseline (p = 0.03), but not different from PLA (baseline: 39 ± 6, CAS: 62 ± 8, PLA: 55 ± 5 mm). CAS consumption before sleep did not affect fat or glucose metabolism, REE, or suppress appetite in hyperinsulemic obese men. CAS may be consumed before sleep without impeding overnight or morning fat metabolism in young, obese men.


Robust inference in discrete hazard models for randomized clinical trials.
Time-to-event data in which failures are only assessed at discrete time points are common in many clinical trials. Examples include oncology studies where events are observed through periodic screenings such as radiographic scans. When the survival endpoint is acknowledged to be discrete, common methods for the analysis of observed failure times include the discrete hazard models (e.g., the discrete-time proportional hazards and the continuation ratio model) and the proportional odds model. In this manuscript, we consider estimation of a marginal treatment effect in discrete hazard models where the constant treatment effect assumption is violated. We demonstrate that the estimator resulting from these discrete hazard models is consistent for a parameter that depends on the underlying censoring distribution. An estimator that removes the dependence on the censoring mechanism is proposed and its asymptotic distribution is derived. Basing inference on the proposed estimator allows for statistical inference that is scientifically meaningful and reproducible. Simulation is used to assess the performance of the presented methodology in finite samples.


Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms.
Candida albicans is implicated in many biomaterial-related infections. Typically, these infections are associated with biofilm formation. Cells in biofilms display phenotypic traits that are dramatically different from those of their free-floating planktonic counterparts and are notoriously resistant to antimicrobial agents. Consequently, biofilm-related infections are inherently difficult to treat and to fully eradicate with normal treatment regimens. Here, we report a rapid and highly reproducible microtiter-based colorimetric assay for the susceptibility testing of fungal biofilms, based on the measurement of metabolic activities of the sessile cells by using a formazan salt reduction assay. The assay was used for in vitro antifungal susceptibility testing of several C. albicans strains grown as biofilms against amphotericin B and fluconazole and the increased resistance of C. albicans biofilms against these antifungal agents was demonstrated. Because of its simplicity, compatibility with a widely available 96-well microplate platform, high throughput, and automation potential, we believe this assay represents a promising tool for the standardization of in vitro antifungal susceptibility testing of fungal biofilms.


A comparative study of salt tolerance parameters in 11 wild relatives of Arabidopsis thaliana.
Salinity is an abiotic stress that limits both yield and the expansion of agricultural crops to new areas. In the last 20 years our basic understanding of the mechanisms underlying plant tolerance and adaptation to saline environments has greatly improved owing to active development of advanced tools in molecular, genomics, and bioinformatics analyses. However, the full potential of investigative power has not been fully exploited, because the use of halophytes as model systems in plant salt tolerance research is largely neglected. The recent introduction of halophytic Arabidopsis-Relative Model Species (ARMS) has begun to compare and relate several unique genetic resources to the well-developed Arabidopsis model. In a search for candidates to begin to understand, through genetic analyses, the biological bases of salt tolerance, 11 wild relatives of Arabidopsis thaliana were compared: Barbarea verna, Capsella bursa-pastoris, Hirschfeldia incana, Lepidium densiflorum, Malcolmia triloba, Lepidium virginicum, Descurainia pinnata, Sisymbrium officinale, Thellungiella parvula, Thellungiella salsuginea (previously T. halophila), and Thlaspi arvense. Among these species, highly salt-tolerant (L. densiflorum and L. virginicum) and moderately salt-tolerant (M. triloba and H. incana) species were identified. Only T. parvula revealed a true halophytic habitus, comparable to the better studied Thellungiella salsuginea. Major differences in growth, water transport properties, and ion accumulation are observed and discussed to describe the distinctive traits and physiological responses that can now be studied genetically in salt stress research.


In my day II: Reflecting on the transformative potential of incorporating celebrations into the nursing curriculum.
Understanding one's history is a powerful way to build a sense of belonging, identity and connection. Similarly, history learning can be a powerful component in the core curriculum for undergraduate nursing. History learning develops thinking skills transferable to and necessary in nursing practice. Additionally, awareness about the profession's struggles, achievements and enduring concerns is raised and belief that an individual or group can have influence is affirmed. Perseverance, commitment and seeing the big picture gives a nurse's career meaning and purpose. All of these factors can produce a transformed perspective in today's learners, who are often present-focused, isolated and disconnected from the past and the profession. This paper reports an evaluation of a second interactive learning experience held at the University of the Sunshine Coast to celebrate International Nurses Day 2010. In a previous paper, we shared our initial insights after the success of the first event, and now build upon those insights by examining the transformative learning provoked by the experience, from the points of view of students and staff.


Bathing-suit ichthyosis. A peculiar phenotype of lamellar ichthyosis in South African blacks.
Autosomal recessive congenital ichthyosis is a heterogeneous group of disorders of cornification showing a wide spectrum of clinical phenotypes.Here, thirteen cases of a so far unreported distinct type of autosomal recessive congenital ichthyosis are described. All the skin lesions were present at birth and ten were then compatible with collodion baby. Shedding of the collodion led to an ichthyotic condition with a very characteristic distribution of the lesions which involved the trunk, the most proximal parts of the upper limbs, scalp and neck but spared the central face and extremities. The term "bathing-suit" ichthyosis is suggested for this unusual phenotype of lamellar ichthyosis.