Asymmetry of the budding yeast Tem1 GTPase at spindle poles is required for spindle positioning but not for mitotic exit.
The asymmetrically dividing yeast S. cerevisiae assembles a bipolar spindle well after establishing the future site of cell division (i.e., the bud neck) and the division axis (i.e., the mother-bud axis). A surveillance mechanism called spindle position checkpoint (SPOC) delays mitotic exit and cytokinesis until the spindle is properly positioned relative to the mother-bud axis, thereby ensuring the correct ploidy of the progeny. SPOC relies on the heterodimeric GTPase-activating protein Bub2/Bfa1 that inhibits the small GTPase Tem1, in turn essential for activating the mitotic exit network (MEN) kinase cascade and cytokinesis. The Bub2/Bfa1 GAP and the Tem1 GTPase form a complex at spindle poles that undergoes a remarkable asymmetry during mitosis when the spindle is properly positioned, with the complex accumulating on the bud-directed old spindle pole. In contrast, the complex remains symmetrically localized on both poles of misaligned spindles. The mechanism driving asymmetry of Bub2/Bfa1/Tem1 in mitosis is unclear. Furthermore, whether asymmetry is involved in timely mitotic exit is controversial. We investigated the mechanism by which the GAP Bub2/Bfa1 controls GTP hydrolysis on Tem1 and generated a series of mutants leading to constitutive Tem1 activation. These mutants are SPOC-defective and invariably lead to symmetrical localization of Bub2/Bfa1/Tem1 at spindle poles, indicating that GTP hydrolysis is essential for asymmetry. Constitutive tethering of Bub2 or Bfa1 to both spindle poles impairs SPOC response but does not impair mitotic exit. Rather, it facilitates mitotic exit of MEN mutants, likely by increasing the residence time of Tem1 at spindle poles where it gets active. Surprisingly, all mutant or chimeric proteins leading to symmetrical localization of Bub2/Bfa1/Tem1 lead to increased symmetry at spindle poles of the Kar9 protein that mediates spindle positioning and cause spindle misalignment. Thus, asymmetry of the Bub2/Bfa1/Tem1 complex is crucial to control Kar9 distribution and spindle positioning during mitosis.


ShopSmart 4 Health: results of a randomized controlled trial of a behavioral intervention promoting fruit and vegetable consumption among socioeconomically disadvantaged women.
Behavioral interventions show potential for promoting increased fruit and vegetable consumption in the general population. However, little is known about their effectiveness or cost-effectiveness among socioeconomically disadvantaged groups, who are less likely to consume adequate fruit and vegetables. This study investigated the effects and costs of a behavior change intervention for increasing fruit and vegetable purchasing and consumption among socioeconomically disadvantaged women. ShopSmart 4 Health was a randomized controlled trial involving a 3-mo retrospective baseline data collection phase [time (T) 0], a 6-mo intervention (T1-T2), and a 6-mo no-intervention follow-up (T3). Socioeconomically disadvantaged women who were primary household shoppers in Melbourne, Australia, were randomly assigned to either a behavior change intervention arm (n = 124) or a control arm (n = 124). Supermarket transaction (sales) data and surveys measured the main outcomes: fruit and vegetable purchases and self-reported fruit and vegetable consumption. An analysis of supermarket transaction data showed no significant intervention effects on vegetable or fruit purchasing at T2 or T3. Participants in the behavior change intervention arm reported consumption of significantly more vegetables during the intervention (T2) than did controls, with smaller intervention effects sustained at 6 mo postintervention (T3). Relative to controls, vegetable consumption increased by ∼0.5 serving · participant(-1) · d(-1) from baseline to T2 and remained 0.28 servings/d higher than baseline at T3 among those who received the intervention. There was no intervention effect on reported fruit consumption. The behavior change intervention cost A$3.10 (in Australian dollars) · increased serving of vegetables(-1) · d(-1)CONCLUSIONS: This behavioral intervention increased vegetable consumption among socioeconomically disadvantaged women. However, the lack of observed effects on fruit consumption and on both fruit and vegetable purchasing at intervention stores suggests that further investigation of effective nutrition promotion approaches for this key target group is required. The ShopSmart 4 Health trial was registered at www.isrctn.com as ISRCTN48771770.


Construction of the basic skeleton of ophiobolin A and variecolin.
Ophiobolin and variecolin type sesterterpenoids belong to cyclooctane-containing natural products. Both sesterterpenoids have challenging structures and appealing biological activities. We envisioned that a key tandem ring closing metathesis of dienynes could provide the basic skeleton of ophiobolin A and variecolin. We report herein the detailed reactivities of the dienynes which furnished the 5-8-5 and 5-8-6-5 rings efficiently.


Genetic relationships among mastitis and alternative somatic cell count traits in the first 3 lactations of Swedish Holsteins.
The objectives of this study were to estimate heritabilities of, and genetic correlations among, clinical mastitis (CM), subclinical mastitis (SCM), and alternative somatic cell count (SCC) traits in the first 3 lactations of Swedish Holstein cows, and to estimate genetic correlations for the alternative traits across lactations. Data from cows having their first calving between 2002 and 2009 were used. The alternative SCC traits were based on information on CM and monthly test-day (TD) records of SCC traits of 178,613, 116,079, and 64,474 lactations in first, second, or third parity, respectively. Sires had an average of 230, 165, or 124 daughters in the data (parities 1, 2, or 3, respectively). Subclinical mastitis was defined as the number of periods with an SCC >150,000 cell/mL and without a treatment for CM. Average TD SCC between 5 and 150 d was used as a reference trait. The alternative SCC traits analyzed were 1) presence of at least 1 TD SCC between 41,000 and 80,000 cell/mL (TD41-80), 2) at least 1 TD SCC >500,000 cells/mL, 3) standard deviation of log SCC over the lactation, 4) number of infection peaks, and 5) average days diseased per peak. The same variables in different parities were treated as distinct traits. The statistical model considered the effects of herd-year, year, month, age at calving, animal, and residual. Heritability estimates were 0.07 to 0.08 for CM, 0.12 to 0.17 for SCM, and 0.14 for SCC150. For the alternative traits, heritability estimates were 0.12 to 0.17 for standard deviation of log SCC, TD SCC >500,000 cells/mL, and average days diseased per peak, and 0.06 to 0.10 for TD41-80 and number of infection peaks. Genetic correlations between CM with SCM were 0.62 to 0.74, and correlations for these traits with the alternative SCC traits were positive and very high (0.67 to 0.82 for CM, and 0.94 to 0.99 for SCM). Trait TD41-80 was the only alternative trait that showed negative, favorable, genetic correlations with CM (-0.22 to -0.50) and SCM (-0.48 to -0.85) because it is associated with healthy cows. Genetic correlations among the alternative traits in all 3 parities were high (0.93 to 0.99, 0.92 to 0.98, and 0.78 to 0.99, respectively). The only exception was TD41-80, which showed moderate to strong negative correlations with the rest of the traits. Genetic correlations of the same trait across parities were in general positive and very high (0.83 to 0.99). In conclusion, these alternative SCC traits could be used in practical breeding programs aiming to improve udder health in dairy cattle.


Three-Dimensional Simulations of Tearing and Intermittency in Coronal Jets.
Observations of coronal jets increasingly suggest that local fragmentation and intermittency play an important role in the dynamics of these events. In this work we investigate this fragmentation in high-resolution simulations of jets in the closed-field corona. We study two realizations of the embedded-bipole model, whereby impulsive helical outflows are driven by reconnection between twisted and untwisted field across the domed fan plane of a magnetic null. We find that the reconnection region fragments following the onset of a tearing-like instability, producing multiple magnetic null points and flux-rope structures within the current layer. The flux ropes formed within the weak-field region in the center of the current layer are associated with "blobs" of density enhancement that become filamentary threads as the flux ropes are ejected from the layer, whereupon new flux ropes form behind them. This repeated formation and ejection of flux ropes provides a natural explanation for the intermittent outflows, bright blobs of emission, and filamentary structure observed in some jets. Additional observational signatures of this process are discussed. Essentially all jet models invoke reconnection between regions of locally closed and locally open field as the jet-generation mechanism. Therefore, we suggest that this repeated tearing process should occur at the separatrix surface between the two flux systems in all jets. A schematic picture of tearing-mediated jet reconnection in three dimensions is outlined.


SupPoRtive Exercise Programmes for Accelerating REcovery after major ABdominal Cancer surgery trial (PREPARE-ABC): Study protocol for a multicentre randomized controlled trial.
Exercise programmes can increase cardiopulmonary reserve and functional capacity prior to surgery and can improve clinical, functional and survival outcomes after a colorectal cancer diagnosis. However, the impact of pre- and postoperative exercise on postoperative recovery outcomes and longer-term health-related quality of life are unknown; thus, there is a need for high-quality randomized controlled trials. SupPoRtive Exercise Programmes for Accelerating REcovery after major Abdominal Cancer surgery (PREPARE-ABC) is a three-arm multicentre randomized controlled trial with internal pilot. The primary objective is to assess the effects of pre- and postoperative exercise on surgical outcomes and longer-term health-related quality of life in cancer patients undergoing colorectal resection. The aim of PREPARE-ABC is to randomize 1146 patients at the individual level (1:1:1) to hospital-supervised exercise, home-supported exercise or treatment as usual. The primary outcomes are short-term (30-day) morbidity, assessed using the Clavien-Dindo classification, and longer-term health-related quality of life, assessed using the Medical Outcomes Study Health Questionnaire (36-item Short-Form Survey [SF-36]). Secondary outcomes include cardiopulmonary fitness, physical activity behaviour change, psychological health status and cost-effectiveness. A process evaluation of intervention delivery and usual care also will be undertaken. This is the first UK-based definitive randomized controlled trial to investigate the effects of pre- and postoperative exercise on short-term postoperative health outcomes and longer-term health-related quality of life in colorectal cancer patients. The trial will yield robust clinical and cost-effectiveness data to underpin clinical guidance on how exercise programmes should be implemented in the routine management of patients undergoing major colorectal cancer surgery.


Age-related sperm DNA methylation changes are transmitted to offspring and associated with abnormal behavior and dysregulated gene expression.
Advanced paternal age (APA) has been shown to be a significant risk factor in the offspring for neurodevelopmental psychiatric disorders, such as schizophrenia and autism spectrum disorders. During aging, de novo mutations accumulate in the male germline and are frequently transmitted to the offspring with deleterious effects. In addition, DNA methylation during spermatogenesis is an active process, which is susceptible to errors that can be propagated to subsequent generations. Here we test the hypothesis that the integrity of germline DNA methylation is compromised during the aging process. A genome-wide DNA methylation screen comparing sperm from young and old mice revealed a significant loss of methylation in the older mice in regions associated with transcriptional regulation. The offspring of older fathers had reduced exploratory and startle behaviors and exhibited similar brain DNA methylation abnormalities as observed in the paternal sperm. Offspring from old fathers also had transcriptional dysregulation of developmental genes implicated in autism and schizophrenia. Our findings demonstrate that DNA methylation abnormalities arising in the sperm of old fathers are a plausible mechanism to explain some of the risks that APA poses to resulting offspring.


Low-Contrast Detail Phantom: Proof of Concept.
To investigate the concept of filling the air gaps of the conventional contrast detail phantom (CDP) with various concentrations of contrast media, and to develop a variable level of attenuation-level differential phantoms that could be more appropriate for contrast measurements in some radiology cases. Images were acquired using the digital radiography system of the traditional CDP (Perspex/air hole phantom) and the novel form of CDP where the air holes were replaced with attenuating material. In this study, two different attenuating materials were introduced, water and a 30% concentration of iodine-based contrast medium. Image quality was assessed using automated processing to calculate the image quality factor (IQF)inv. Phantom studies indicate that lower contrast levels are obtained when CDP holes are filled with water and a 30% concentration of iodine contrast media than those observed for air/Perspex or traditional CDP. As an example, when a 5-mAs beam is used the IQFinv values are 5.32 in the case of air filling the holes; however, when these holes are filled with water under the same conditions, the value of the IQFinv drops to 2.55, and to 2.83 when 30% of contrast media is used. Other concentrations were also tested. These results indicate that it is possible to extend the contrast scale in these phantoms to include ranges that are more realistic for a patient's body than just air and tissue-equivalent material. These findings indicate that the proposed extension of the contrast scales allows smaller changes in contrast to be discerned. This is due to the small attenuation differences of the subject materials (e.g, 30% contrast liquid and wax) from the traditional form of CDP (material/air). This suggests that the low form of the CDP may have a useful role in assessing image quality in planar radiology as an evaluation tool to better represent low-subject contrast imaging requirements.


A new method for the in vivo volumetric measurement of the human hippocampus with high neuroanatomical accuracy.
Accurate and reproducible in vivo measurement of hippocampal volumes using magnetic resonance (MR) imaging is complicated by the morphological complexity of the structure. Additionally, separation of certain parts of the hippocampus from the adjacent brain structures on MR images is sometimes very difficult. These difficulties have led most investigators to either use arbitrary landmarks or to exclude certain parts of the structure from their measurements. Based on three-dimensional MR data, we have developed a reliable in vivo volumetric measurement of the human hippocampus. In contrast to most of the previously described volumetric MR-based methods, we aimed to sample the entire hippocampal formation using its true anatomical definition. This was accomplished by relying on the capacity of the BRAINS software to simultaneously visualize in multiple planes, to "telegraph" tracings or cursor position from one plane to another, and to simultaneously rely on multispectral data from three different image sets (T1, T2, and tissue classified). The methods for identifying boundaries and measuring the hippocampal volume are described. The method has excellent reliability, sensitivity, and specificity. The method may be of use in studies of structure-function relationships in neuropsychiatric disorders such as schizophrenia, temporal lobe epilepsy, and Alzheimer's disease. Future work will use these measurements as training data for a neural net-based technique to identify the anatomical boundaries automatically.


NaV Channels: Assaying Biosynthesis, Trafficking, Function.
Integral to the cell surface is channels, pumps, and exchanger proteins that facilitate the movement of ions across the membrane. Ion channels facilitate the passive movement of ions down an electrochemical gradient. Ion pumps actively use energy to actively translocate ions, often against concentration or voltage gradients, while ion exchangers utilize energy to couple the transport of different ion species such that one ion moves down its gradient and the released free energy is used to drive the movement of a different ion against its electrochemical gradient. Some ion pumps and exchangers may be electrogenic, i.e., the ion transport they support is not electrically neutral and generates a current. Functions of these pore-forming membrane proteins include the establishment of membrane potentials, gating of ions flows across the cell membrane to elicit action potentials and other electrical signals, as well as the regulation of cell volumes. The major forms of ion channels include voltage-, ligand-, and signal-gated channels. In this review, we describe mammalian voltage dependent Na (NaV) channels.


Genetic analysis of variation in human meiotic recombination.
The number of recombination events per meiosis varies extensively among individuals. This recombination phenotype differs between female and male, and also among individuals of each gender. In this study, we used high-density SNP genotypes of over 2,300 individuals and their offspring in two datasets to characterize recombination landscape and to map the genetic variants that contribute to variation in recombination phenotypes. We found six genetic loci that are associated with recombination phenotypes. Two of these (RNF212 and an inversion on chromosome 17q21.31) were previously reported in the Icelandic population, and this is the first replication in any other population. Of the four newly identified loci (KIAA1462, PDZK1, UGCG, NUB1), results from expression studies provide support for their roles in meiosis. Each of the variants that we identified explains only a small fraction of the individual variation in recombination. Notably, we found different sequence variants associated with female and male recombination phenotypes, suggesting that they are regulated by different genes. Characterization of genetic variants that influence natural variation in meiotic recombination will lead to a better understanding of normal meiotic events as well as of non-disjunction, the primary cause of pregnancy loss.


Exercise training in pediatric patients with end-stage renal disease.
The objective of this study was to determine the feasibility and efficacy of an exercise training program to improve exercise capacity and fatigue level in pediatric patients with end-stage renal disease (ESRD). Twenty children on dialysis intended to perform a 12-week graded community-based exercise program. Exercise capacity and fatigue level were studied; muscle force and health-related quality of life were secondary outcomes. All outcomes were measured at baseline (T = 0) and after intervention (T = 1). Fourteen of the 20 patients (70%) either did not start the program or did not complete the program. Of these patients, seven did not complete or even start the exercise program because of a combination of lack of time and motivational problems. Six patients were not able to continue the program or were unable to do the follow-up measurements because of medical problems. Exercise capacity and muscle strength was higher after the exercise program in the children who completed the training. In conclusion, exercise training is difficult to perform in children with ESRD and is not always feasible in real-life situations for many children with ESRD.


Comparison of reoperation rates following ankle arthrodesis and total ankle arthroplasty.
The role of ankle arthroplasty in the treatment of ankle arthritis is controversial. Ankle fusion is commonly performed, but there is ongoing concern about functional limitations and arthritis in the adjacent subtalar joint following ankle arthrodesis. The use of ankle arthroplasty as an alternative to ankle fusion is expanding, but reported results have been limited to those in case series. The purpose of this study was to compare the reoperation rates following ankle arthrodesis and ankle replacement on the basis of observational, population-based data from all inpatient admissions in California over a ten-year period. Our hypothesis was that patients treated with ankle replacement would have a lower risk of undergoing subtalar fusion but a higher overall risk of undergoing major revision surgery. We used California's hospital discharge database to identify patients who had undergone ankle replacement or ankle arthrodesis as inpatients in the years 1995 through 2004. Short-term outcomes, including rates of major revision surgery, pulmonary embolism, amputation, and infection, were examined. Long-term outcomes that were analyzed included the rates of major revision surgery and subtalar joint fusion. Logistic and proportional hazard regression models were used to estimate the impact of the choice of ankle replacement or ankle fusion on the rates of adverse outcomes, with adjustment for patient factors including age and comorbidity. A total of 4705 ankle fusions and 480 ankle replacements were performed during the ten-year study period. Patients who had undergone ankle replacement had an increased risk of device-related infection and of having a major revision procedure. The rates of major revision surgery after ankle replacement were 9% at one year and 23% at five years compared with 5% and 11% following ankle arthrodesis. Patients treated with ankle arthrodesis had a higher rate of subtalar fusion at five years postoperatively (2.8%) than did those treated with ankle replacement (0.7%). Regression analysis confirmed a significant increase in the risk of major revision surgery (hazard ratio, 1.93 [95% confidence interval, 1.50 to 2.49]; p < 0.001) but a decreased risk of subtalar fusion (hazard ratio, 0.28 [95% confidence interval, 0.09 to 0.87]; p = 0.03) in patients treated with ankle replacement compared with those treated with ankle fusion. This study confirms that, compared with ankle fusion, ankle replacement is associated with a higher risk of complications but also potential advantages in terms of a decreased risk of the patient requiring subtalar joint fusion. Additional controlled trials are needed to clarify the appropriate indications for ankle arthrodesis and ankle replacement.


Alignment of the stellar spin with the orbits of a three-planet system.
The Sun's equator and the planets' orbital planes are nearly aligned, which is presumably a consequence of their formation from a single spinning gaseous disk. For exoplanetary systems this well-aligned configuration is not guaranteed: dynamical interactions may tilt planetary orbits, or stars may be misaligned with the protoplanetary disk through chaotic accretion , magnetic interactions or torques from neighbouring stars. Indeed, isolated 'hot Jupiters' are often misaligned and even orbiting retrograde. Here we report an analysis of transits of planets over starspots on the Sun-like star Kepler-30 (ref. 8), and show that the orbits of its three planets are aligned with the stellar equator. Furthermore, the orbits are aligned with one another to within a few degrees. This configuration is similar to that of our Solar System, and contrasts with the isolated hot Jupiters. The orderly alignment seen in the Kepler-30 system suggests that high obliquities are confined to systems that experienced disruptive dynamical interactions. Should this be corroborated by observations of other coplanar multi-planet systems, then star-disk misalignments would be ruled out as the explanation for the high obliquities of hot Jupiters, and dynamical interactions would be implicated as the origin of hot Jupiters.


Probiotics supplementation during pregnancy or infancy for the prevention of atopic dermatitis: a meta-analysis.
The study of probiotics to prevent allergic conditions has yielded conflicting results in children. We undertook a meta-analysis of randomized controlled trials to investigate whether probiotic use during pregnancy and early life decreases the incidence of atopic dermatitis and immunoglobulin E (IgE)-associated atopic dermatitis in infants and young children. We performed a systematic literature search in Medline, Embase, and Cochrane Library, updated to October 2011. The intervention was diet supplementation with probiotics versus placebo. Primary outcomes were incidence of atopic dermatitis and IgE-associated atopic dermatitis. We calculated summary relative risks (RRs) and corresponding 95% confidence intervals (CIs), using both fixed- and random-effects models. We computed summary estimates across several strata, including study period, type of patient, dose, and duration of intervention, and we assessed the risk of bias within and across trials. We identified 18 publications based on 14 studies. Meta-analysis demonstrated that probiotic use decreased the incidence of atopic dermatitis (RR = 0.79 [95% CI = 0.71-0.88]). Studies were fairly homogeneous (I = 24.0%). The corresponding RR of IgE-associated atopic dermatitis was 0.80 (95% CI = 0.66-0.96). No appreciable difference emerged across strata, nor was there evidence of publication bias. This meta-analysis provided evidence in support of a moderate role of probiotics in the prevention of atopic dermatitis and IgE-associated atopic dermatitis in infants. The favorable effect was similar regardless of the time of probiotic use (pregnancy or early life) or the subject(s) receiving probiotics (mother, child, or both).


Advanced methylome analysis after bisulfite deep sequencing: an example in Arabidopsis.
Deep sequencing after bisulfite conversion (BS-Seq) is the method of choice to generate whole genome maps of cytosine methylation at single base-pair resolution. Its application to genomic DNA of Arabidopsis flower bud tissue resulted in the first complete methylome, determining a methylation rate of 6.7% in this tissue. BS-Seq reads were mapped onto an in silico converted reference genome, applying the so-called 3-letter genome method. Here, we present BiSS (Bisufite Sequencing Scorer), a new method applying Smith-Waterman alignment to map bisulfite-converted reads to a reference genome. In addition, we introduce a comprehensive adaptive error estimate that accounts for sequencing errors, erroneous bisulfite conversion and also wrongly mapped reads. The re-analysis of the Arabidopsis methylome data with BiSS mapped substantially more reads to the genome. As a result, it determines the methylation status of an extra 10% of cytosines and estimates the methylation rate to be 7.7%. We validated the results by individual traditional bisulfite sequencing for selected genomic regions. In addition to predicting the methylation status of each cytosine, BiSS also provides an estimate of the methylation degree at each genomic site. Thus, BiSS explores BS-Seq data more extensively and provides more information for downstream analysis.


Modulated-alignment dual-axis (MAD) confocal microscopy for deep optical sectioning in tissues.
A strategy is presented to enable optical-sectioning microscopy with improved contrast and imaging depth using low-power (0.5 - 1 mW) diode laser illumination. This technology combines the inherent strengths of focal-modulation microscopy and dual-axis confocal (DAC) microscopy for rejecting out-of-focus and multiply scattered background light in tissues. The DAC architecture is unique in that it utilizes an intersecting pair of illumination and collection beams to improve the spatial-filtering and optical-sectioning performance of confocal microscopy while focal modulation selectively 'labels' in-focus signals via amplitude modulation. Simulations indicate that modulating the spatial alignment of dual-axis beams at a frequency f generates signals from the focal volume of the microscope that are modulated at 2f with minimal modulation of background signals, thus providing nearly an order-of-magnitude improvement in optical-sectioning contrast compared to DAC microscopy alone. Experiments show that 2f lock-in detection enhances contrast and imaging depth within scattering phantoms and fresh tissues.


Biomarkers in assessing residential insecticide exposures during pregnancy and effects on fetal growth.
The Columbia Center for Children's Environmental Health is using a combination of environmental and biologic measures to evaluate the effects of prenatal insecticide exposures among urban minorities in New York City. Of the 571 women enrolled, 85% report using some form of pest control during pregnancy and 46% report using exterminators, can sprays, and/or pest bombs. Chlorpyrifos, diazinon, and propoxur were detected in 99.7-100% of 48-h personal air samples collected from the mothers during pregnancy (n = 394) and in 39-70% of blood samples collected from the mothers (n = 326) and/or newborns (n = 341) at delivery. Maternal and newborn blood levels are similar and highly correlated (r = 0.4-08, P < 0.001). Levels of insecticides in blood samples and/or personal air samples decreased significantly following the 2000-2001 U.S. Environmental Protection Agency's regulatory actions to phase out residential use of chlorpyrifos and diazinon. Among infants born prior to 1/1/01, birth weight decreased by 67.3 g (95% confidence interval (CI) -116.6 to -17.8, P = 0.008) and birth length decreased by 0.43 centimeters (95% CI, -0.73 to -0.14, P = 0.004) for each unit increase in log-transformed cord plasma chlorpyrifos levels. Combined measures of (ln)cord plasma chlorpyrifos and diazinon (adjusted for relative potency) were also inversely associated with birth weight and length (P </= 0.007). Birth weight averaged 215.1 g less (95% CI -384.7 to -45.5) among those with the highest exposures compared to those without detectable levels. No association was seen between birth weight and length and cord plasma chlorpyrifos or diazinon among newborns born after 1/1/01 (P > 0.8). Results support recent regulatory action to phase out residential uses of these insecticides.


A prospective longitudinal model predicting early adult alcohol problems: evidence for a robust externalizing pathway.
Risk factors for alcohol problems (AP) include biological and environmental factors that are relevant across development. The pathways through which these factors are related, and how they lead to AP, are optimally considered in the context of a comprehensive developmental model. Using data from a prospectively assessed, population-based UK cohort, we constructed a structural equation model that integrated risk factors reflecting individual, family and peer/community-level constructs across childhood, adolescence and young adulthood. These variables were used to predict AP at the age of 20 years. The final model explained over 30% of the variance in liability to age 20 years AP. Most prominent in the model was an externalizing pathway to AP, with conduct problems, sensation seeking, AP at age 17.5 years and illicit substance use acting as robust predictors. In conjunction with these individual-level risk factors, familial AP, peer relationships and low parental monitoring also predicted AP. Internalizing problems were less consistently associated with AP. Some risk factors previously identified were not associated with AP in the context of this comprehensive model. The etiology of young adult AP is complex, influenced by risk factors that manifest across development. The most prominent pathway to AP is via externalizing and related behaviors. These findings underscore the importance of jointly assessing both biologically influenced and environmental risk factors for AP in a developmental context.


Long-term forest soil warming alters microbial communities in temperate forest soils.
Soil microbes are major drivers of soil carbon cycling, yet we lack an understanding of how climate warming will affect microbial communities. Three ongoing field studies at the Harvard Forest Long-term Ecological Research (LTER) site (Petersham, MA) have warmed soils 5°C above ambient temperatures for 5, 8, and 20 years. We used this chronosequence to test the hypothesis that soil microbial communities have changed in response to chronic warming. Bacterial community composition was studied using Illumina sequencing of the 16S ribosomal RNA gene, and bacterial and fungal abundance were assessed using quantitative PCR. Only the 20-year warmed site exhibited significant change in bacterial community structure in the organic soil horizon, with no significant changes in the mineral soil. The dominant taxa, abundant at 0.1% or greater, represented 0.3% of the richness but nearly 50% of the observations (sequences). Individual members of the Actinobacteria, Alphaproteobacteria and Acidobacteria showed strong warming responses, with one Actinomycete decreasing from 4.5 to 1% relative abundance with warming. Ribosomal RNA copy number can obfuscate community profiles, but is also correlated with maximum growth rate or trophic strategy among bacteria. Ribosomal RNA copy number correction did not affect community profiles, but rRNA copy number was significantly decreased in warming plots compared to controls. Increased bacterial evenness, shifting beta diversity, decreased fungal abundance and increased abundance of bacteria with low rRNA operon copy number, including Alphaproteobacteria and Acidobacteria, together suggest that more or alternative niche space is being created over the course of long-term warming.


Empathy, effectiveness and donations to charity: social psychology's contribution.
Charity organizations often use mailed requests to solicit donations from the public. This is not an efficient way to raise large amounts of money. The challenge addressed in this study was to use social psychology's knowledge of helping processes to make mailed requests more effective. Two constructs were identified as possibly useful: empathy and perceived effectiveness of helping. These were manipulated in a field experiment in a 2 x 2 x 2 factorial design (two levels of empathy, two of need extent, and two of need persistence--the last two factors operationalized perceived effectiveness). Letter soliciting donations to a well-known charity were mailed to a random sample of 2648 people in Perth, Western Australia. Manipulations of the three variables were embedded in the letters. The two effectiveness manipulations produced significant main effects, whereas the empathy manipulation was ineffective. We argue that social psychology's knowledge of helping processes is too confined to narrow, theoretical, laboratory-based phenomena to be directly and immediately applicable to the practices of charities.


Range for normal body temperature in hemodialysis patients and its comparison with that of healthy individuals.
Patients with chronic kidney disease undergoing hemodialysis have an altered homeostasis leading to altered body temperatures. We aimed to determine the range for normal body temperature in hemodialysis patients and compared it to healthy individuals. Also, we determined how much axillary temperatures differed from oral temperatures in both groups and whether axillary temperature is affected by the presence of an arteriovenous fistula (AVF) in hemodialysis patients. Oral and axillary (left & right) temperatures were recorded using an ordinary mercury-in-glass thermometer in 400 subjects (200 hemodialysis patients, 200 healthy individuals) at the Sindh Institute of Urology and Transplantation from mid-May to mid-June 2006. Comparisons were made between the temperatures of both groups. Mean oral temperature in hemodialysis patients was higher than in healthy individuals [98.7 degrees F (37 degrees C) vs. 98.4 degrees F (36.8 degrees C); p < 0.001], as was the mean average axillary temperature [97.7 degrees F (36.5 degrees C) vs. 97.5 degrees F (36.3 degrees C); p = 0.02] and mean left axillary temperature [97.9 degrees F (36.6 degrees C) vs. 97.6 degrees F (36.4 degrees C); p < 0.001]. The fistula arm had higher axillary temperature in 77 (44%) hemodialysis patients. The difference between oral and axillary temperatures varied widely, making it impossible to obtain an accurate correction factor in both groups. Hemodialysis patients have higher normal body temperatures than healthy individuals. Axillary temperatures require cautious interpretation. In hemodialysis patients, the non-fistula arm should be preferred for recording axillary temperatures, as the presence of AVF may cause discrepancies in temperature measurements.


Individual pathogens, pathogen burden and markers of subclinical atherosclerosis: the Multi-Ethnic Study of Atherosclerosis.
We examined the cross-sectional relationships of subclinical atherosclerosis - expressed by carotid intimal-medial thickness and coronary calcification - with antibodies to Chlamydia pneumoniae, Helicobacter pylori, cytomegalovirus, herpes simplex virus, hepatitis A virus, and pathogen burden (number of positive pathogens). A random sample of 1056 individuals chosen from 5030 Multi-Ethnic Study of Atherosclerosis cohort participants were included. After multiple adjustment, no associations were found between atherosclerosis measures and either individual pathogens or pathogen burden. Interactions with inflammatory and endothelial function markers, demographic factors, BMI, high-density lipoprotein, diabetes, and smoking were also explored. The only interaction that was large, qualitative, statistically significant (P < 0.05) and in the expected direction was that between hepatitis A virus and soluble intercellular adhesion molecule-1 with regard to Agatston calcium score: the difference between hepatitis A virus-positive and hepatitis A virus-negative participants was -186 units in participants with soluble intercellular adhesion molecule-1 below the median, and +162 units in those with soluble intercellular adhesion molecule-1 equal or above the median. However, given the number of interactions that were explored, these results must be interpreted cautiously. Findings from the present analyses do not provide support for an infectious etiology for subclinical atherosclerosis. However, the study's limitations, which include its cross-sectional design and insufficient statistical power, suggest that inferences from its findings should be made cautiously.


SU-E-I-89: Applying Image Gently and Image Wisely in Nuclear Medicine.
To demonstrate how principles and recommendations of the Image Gently® and Image Wisely™ campaigns can be clinically implemented. Implementation of Image Gently® and Image Wisely™ guidelines was a quality and safety improvement project for our Nuclear Medicine department in 2011. Pediatric administered radiopharmaceutical doses were compared to recommended doses in the North American Consensus Guidelines for AdministeredRadiopharmaceutical Activities in Children and Adolescents and the European Association of Nuclear Medicine Paediatric Dose Card. Adult administered radiopharmaceutical doses were compared to administered doses in NCRP Report No. 160 and IAEA Report No. 40. Evaluation of site administered radiopharmaceutical doses showed that nearly all doses were comparable to those recommended by national and international advisory bodies. Administered doses for the pediatric renogram and the adult gastric emptying were adjusted to match the recommendations and resultant image quality evaluated. Radiation dose from nuclear medicine procedures should not be overlooked in the focus on CT dose reduction. An estimated 19.7 million nuclear medicine procedures are done annually in the U.S., with doses comparable to that from CT scans. Nuclear medicine departments should evaluate their pediatric administeredradiopharmaceutical doses against the North American Consensus Guidelines for Administered Radiopharmaceutical Activities in Children and Adolescents and evaluate their adult doses against national and international standards. Administered doses that are not comparable to the recommended values should be adjusted accordingly. Additional steps to reduce patient radiation dose include decision support to reduce inappropriate ordering, technique optimization for the CT scan portion of SPECT/CT and PET/CT, use of vendor's dose reduction camera and software technology, use of shorter lived radiopharmaceuticals, and 'right sizing' patient doses by weight.


Release of severe acute respiratory syndrome coronavirus nuclear import block enhances host transcription in human lung cells.
The severe acute respiratory syndrome coronavirus accessory protein ORF6 antagonizes interferon signaling by blocking karyopherin-mediated nuclear import processes. Viral nuclear import antagonists, expressed by several highly pathogenic RNA viruses, likely mediate pleiotropic effects on host gene expression, presumably interfering with transcription factors, cytokines, hormones, and/or signaling cascades that occur in response to infection. By bioinformatic and systems biology approaches, we evaluated the impact of nuclear import antagonism on host expression networks by using human lung epithelial cells infected with either wild-type virus or a mutant that does not express ORF6 protein. Microarray analysis revealed significant changes in differential gene expression, with approximately twice as many upregulated genes in the mutant virus samples by 48 h postinfection, despite identical viral titers. Our data demonstrated that ORF6 protein expression attenuates the activity of numerous karyopherin-dependent host transcription factors (VDR, CREB1, SMAD4, p53, EpasI, and Oct3/4) that are critical for establishing antiviral responses and regulating key host responses during virus infection. Results were confirmed by proteomic and chromatin immunoprecipitation assay analyses and in parallel microarray studies using infected primary human airway epithelial cell cultures. The data strongly support the hypothesis that viral antagonists of nuclear import actively manipulate host responses in specific hierarchical patterns, contributing to the viral pathogenic potential in vivo. Importantly, these studies and modeling approaches not only provide templates for evaluating virus antagonism of nuclear import processes but also can reveal candidate cellular genes and pathways that may significantly influence disease outcomes following severe acute respiratory syndrome coronavirus infection in vivo.


The role of antioxidant vitamins in hypertensive disorders of pregnancy.
Preeclampsia (PE) is an important and a leading cause of both maternal morbidity and adverse perinatal outcomes. Despite progress in perinatal medicine for patients with an established diagnosis of PE, a therapeutic approach other than termination of pregnancy was unsuccessful. Women predisposed to PE begin pregnancy with a certain degree of endothelial dysfunction, a lesion that precedes shallow placentation. The proposed sequence of events comprises endothelial dysfunction, defective trophoblast invasion, and consequential impaired placental perfusion, immune maladaptation and inflammation. The possible link between these could be oxidative stress by excessive production of reactive oxygen species coupled with inadequate or overwhelmed antioxidant defense mechanisms. These defense mechanisms, involving antioxidant vitamins and enzyme systems, may restrain the extent of damage caused by oxidative stress. Markers of oxidative stress in women with established PE were confirmed. Accordingly, these findings support an expected beneficial effect of antioxidant therapy in the prevention of PE and other pregnancy-related disorders. Numerous studies have been carried out in order to investigate this possible and simple prophylactic and/or therapeutic approach in prevention of oxidative stress and eventual reduction of PE and its perinatal complications. In this review the role of vitamin antioxidants in prevention and treatment of PE is discussed. Despite the logic behind using antioxidant vitamins, the data, thus far, are at best conflicting.


Biomechanical characterization of human amniotic membrane preparations for ocular surface reconstruction.
To investigate the tensile and elastic properties of both commercially available and experimental human amniotic membrane preparations. Nine preparations of human amniotic membrane were studied. The four dry preparations were untreated (nonirradiated, n = 20), and gamma (n = 25), low-dose (AmbioDry, Okto Ophtho Inc., Costa Mesa, Calif., USA, n = 20) and high-dose (n = 20) electron beam sterilized. The same dry membranes were moistened with balanced salt solution (n = 20, 34, 20 and 20, respectively). The ninth group consisted of thawed medium-frozen amniotic membrane (AmnioGraft, Bio-Tissue Inc., Miami, Fla., USA, n = 20). The membranes were cut into thin strips, loaded on a gram range load sensor, and stretched incrementally to the point of rupture. The modulus of elasticity, displacement until rupture and maximum tolerated stress were recorded and compared. The dry preparations exhibited higher moduli of elasticity when compared with the moist samples, with the low-dose electron beam-irradiated samples having the greatest mean modulus of elasticity overall and maintaining a high modulus of elasticity as a moist sample (p < 0.05). Moist nonirradiated preparations and thawed medium-frozen preparations stretched the farthest before rupture and experienced the greatest mean stresses at the point of rupture. While 3 of 4 membranes had greater stretch when moistened as compared to their dry counterparts, there was no difference in the membrane stiffness between dry and moistened low-dose electron beam-irradiated samples (p > 0.8). Low-dose electron beam-irradiated amnion appeared to maintain desirable elastic characteristics in transition from a dry to rehydrated state and may thus provide an easy-to-manipulate transplant tissue for ocular surface reconstruction. Moist nonirradiated and thawed medium-frozen tissues, however, may provide surgical advantages as they required greater forces to rupture.


Fractal structure of eigenmodes of unstable-cavity lasers.
We show that the eigenmodes of unstable-cavity lasers have fractal structure, in contrast with the well-known stable-cavity eigenmodes. As with all fractals, the dynamic range over which self-similarity holds is limited; in this case the range is set by diffraction, i.e., by the Fresnel number of the resonator. We determine the fractal dimension of the mode profiles and show that it is related to the aperture shape.


Beckman Access versus the Bayer ACS:180 and the Abbott AxSYM cardiac Troponin-I real-time immunoassays: an observational prospective study.
BACKGROUND: Reliability of cardiac troponin-I assays under real-time conditions has not been previously well studied. Most large published cTnI trials have utilized protocols which required the freezing of serum (or plasma) for delayed batch cTnI analysis. We sought to correlate the presence of the acute ischemic coronary syndrome (AICS) to troponin-I values obtained in real-time by three random-mode analyzer immunoassay systems: the Beckman ACCESS (BA), the Bayer ACS:180 (CC) and the Abbott AxSYM (AX). METHODS: This was an observational prospective study at a university tertiary referral center. Serum from a convenience sampling of telemetry patients was analyzed in real-time for troponin-I by either the BA-CC (Arm-1) or BA-AX (Arm-2) assay pairs. Presence of the AICS was determined retrospectively and then correlated with troponin-I results. RESULTS: 100 patients were enrolled in Arm-1 (38 with AICS) and 94 in Arm-2 (48 with AICS). The BA system produced 51% false positives in Arm-1, 44% in Arm-2, with negative predictive values of 92% and 100% respectively. In Arm-1, the BA and the CC assays had sensitivities of 97% and 63% and specificities of 18% and 87%. In Arm-2, the BA and the AX assays had sensitivities of 100% and 83% and specificities of 11% and 78%. CONCLUSIONS: In real-time analysis, the performance of the AxSYM and ACS:180 assay systems produced more accurate troponin-I results than the ACCESS system.


Expression and activation of caspase-6 in human fetal and adult tissues.
Caspase-6 is an effector caspase that has not been investigated thoroughly despite the fact that Caspase-6 is strongly activated in Alzheimer disease brains. To understand the full physiological impact of Caspase-6 in humans, we investigated Caspase-6 expression. We performed western blot analyses to detect the pro-Caspase-6 and its active p20 subunit in fetal and adult lung, kidney, brain, spleen, muscle, stomach, colon, heart, liver, skin, and adrenals tissues. The levels were semi-quantitated by densitometry. The results show a ubiquitous expression of Caspase-6 in most fetal tissues with the lowest levels in the brain and the highest levels in the gastrointestinal system. Caspase-6 active p20 subunits were only detected in fetal stomach. Immunohistochemical analysis of a human fetal embryo showed active Caspase-6 positive apoptotic cells in the dorsal root ganglion, liver, lung, kidney, ovary, skeletal muscle and the intestine. In the adult tissues, the levels of Caspase-6 were lower than in fetal tissues but remained high in the colon, stomach, lung, kidney and liver. Immunohistological analyses revealed that active Caspase-6 was abundant in goblet cells and epithelial cells sloughing off the intestinal lining of the adult colon. These results suggest that Caspase-6 is likely important in most tissues during early development but is less involved in adult tissues. The low levels of Caspase-6 in fetal and adult brain indicate that increased expression as observed in Alzheimer Disease is a pathological condition. Lastly, the high levels of Caspase-6 in the gastrointestinal system indicate a potential specific function of Caspase-6 in these tissues.


Increasing intention to cook from basic ingredients: A randomised controlled study.
The promotion of home cooking is a strategy used to improve diet quality and health. However, modern home cooking typically includes the use of processed food which can lead to negative outcomes including weight gain. In addition, interventions to improve cooking skills do not always explain how theory informed their design and implementation. The Behaviour Change Technique (BCT) taxonomy successfully employed in other areas has identified essential elements for interventions. This study investigated the effectiveness of different instructional modes for learning to cook a meal, designed using an accumulating number of BCTs, on participant's perceived difficulty, enjoyment, confidence and intention to cook from basic ingredients. 141 mothers aged between 20 and 39 years from the island of Ireland were randomised to one of four conditions based on BCTs (1) recipe card only [control condition]; (2) recipe card plus video modelling; (3) recipe card plus video prompting; (4) recipe card plus video elements. Participants rated their enjoyment, perceived difficulty, confidence and intention to cook again pre, mid and post experiment. Repeated one-way factorial ANOVAs, correlations and a hierarchical regression model were conducted. Despite no significant differences between the different conditions, there was a significant increase in enjoyment (P < 0.001), confidence (P < 0.001) and intention to cook from basics again (P < 0.001) and a decrease in perceived difficulty (P = 0.001) after the experiment in all conditions. Intention to cook from basics pre-experiment, and confidence and enjoyment (both pre and post experiment) significantly contributed to the final regression model explaining 42% of the variance in intention to cook from basics again. Cooking interventions should focus on practical cooking and increasing participants' enjoyment and confidence during cooking to increase intention to cook from basic ingredients at home.


Muscle-Bone Interactions in Chinese Men and Women Aged 18-35 Years.
To characterize bone mineral density (BMD), bone strength, muscle and fat mass, and muscle strength and power in Chinese women (n = 25) and men (n = 28) classified as in the bone accrual phase (18-25 years) or in the peak bone mass phase (26-35 years). Calcium intakes, physical activity levels, and serum vitamin D were measured. Dual-energy X-ray absorptiometry (DXA) assessed body composition, lumbar spine, and hip areal BMD (aBMD) variables and peripheral quantitative computed tomography (pQCT) assessed cortical and trabecular volumetric BMD (vBMD) and bone strength. Muscle strength and power were assessed by grip strength, leg press, and vertical jump tests. Calcium, serum vitamin D, and physical activity levels were similar across age and sex groups. Significant sex differences (p < 0.05) were found for most body composition variables, hip aBMD, tibia variables, and muscle strength and power. Adjusting for height and weight eliminated most of the significant sex differences. Women showed stronger positive correlations between body composition and bone variables (r = 0.44 to 0.78) than men. Also, correlations between muscle strength/power were stronger in women vs. men (r = 0.43 to 0.82). Bone traits were better related to body composition and muscle function in Chinese women compared to Chinese men aged 18 to 35 years, and peak bone mass seems to be achieved by 25 years of age in both Chinese men and women since there were no differences between the two age groups.


Molecular Diagnosis of Sexually Transmitted Chlamydia trachomatis in the United States.
Chlamydia, with its Chlamydia trachomatis etiology, is the most common bacterial sexually transmitted infection in the United States and is often transmitted via asymptomatic individuals. This review summarizes traditional and molecular-based diagnostic modalities specific to C. trachomatis. Several commercially available, FDA-approved molecular methods to diagnose urogenital C. trachomatis infection include nucleic acid hybridization, signal amplification, polymerase chain reaction, strand displacement amplification, and transcription-mediated amplification. Molecular-based methods are rapid and reliable genital specimen screening measures, especially when applied to areas of high disease prevalence. However, clinical and analytical sensitivity for some commercial systems decreases dramatically when testing urine samples. In vitro experiments and clinical data suggest that transcription-mediated amplification has greater analytical sensitivity than the other molecular-based methods currently available. This difference may be further exhibited in testing of extragenital specimens from at-risk patient demographics. The development of future molecular testing could address conundrums associated with confirmatory testing, medicolegal testing, and test of cure.


Identifying fecal sources in a selected catchment reach using multiple source-tracking tools.
Given known limitations of current microbial source-tracking (MST) tools, emphasis on small, simple study areas may enhance interpretations of fecal contamination sources in streams. In this study, three MST tools-Escherichia coli repetitive element polymerase chain reaction (rep-PCR), coliphage typing, and Bacteroidales 16S rDNA host-associated markers-were evaluated in a selected reach of Plum Creek in south-central Nebraska. Water-quality samples were collected from six sites. One reach was selected for MST evaluation based on observed patterns of E. coli contamination. Despite high E. coli concentrations, coliphages were detected only once among water samples, precluding their use as a MST tool in this setting. Rep-PCR classification of E. coli isolates from both water and sediment samples supported the hypothesis that cattle and wildlife were dominant sources of fecal contamination, with minor contributions by horses and humans. Conversely, neither ruminant nor human sources were detected by Bacteroidales markers in most water samples. In bed sediment, ruminant- and human-associated Bacteroidales markers were detected throughout the interval from 0 to 0.3 m, with detections independent of E. coli concentrations in the sediment. Although results by E. coli-based and Bacteroidales-based MST methods led to similar interpretations, detection of Bacteroidales markers in sediment more commonly than in water indicates that different tools to track fecal contamination (in this case, tools based on Bacteroidales DNA and E. coli isolates) may have varying relevance to the more specific goal of tracking the sources of E. coli in watersheds. This is the first report of simultaneous, toolbox approach application of a library-based and marker-based MST analyses to flowing surface water.


Estrogen-like effects of tamoxifen on human endometrial carcinoma transplanted into nude mice.
The effect of tamoxifen (TAM) on human endometrial carcinoma was investigated in nude mice bearing an estrogen receptor-positive or estrogen receptor-negative tumor. The receptor-negative tumor grew rapidly, and the rates of tumor growth of 17 beta-estradiol or TAM-treated animals were identical to the rate of controls. The estradiol receptor and progesterone receptor (PR) concentrations in the tumor cytosol remained undetectable under all experimental conditions. In contrast, the rate of growth of steroid receptor positive tumor was significantly accelerated in the presence of TAM compared to controls (p less than 0.02). The increased tumor growth rate was, however, significantly lower (p less than 0.01) than that observed in animals receiving 17 beta-estradiol. The PR concentration in these tumors was elevated in response to TAM treatment. That the TAM-induced PR was indeed functional was evident from (a) increased activities of the progestin-sensitive enzyme, 17 beta-estradiol hydroxysteroid dehydrogenase and (b) histological appearance of subnuclear vacuolization in these tumors after progestin administration. These studies indicate that continuous, short-term administration of TAM to nude mice results in an estrogen-like effect on endometrial carcinoma. Based on the finding that TAM induces functional PR, we predict that steroid receptor-positive endometrial carcinoma may show a greater response rate to combined, long-term treatment with TAM and progestin.


Rab35 controls cilium length, function and membrane composition.
Rab and Arl guanine nucleotide-binding (G) proteins regulate trafficking pathways essential for the formation, function and composition of primary cilia, which are sensory devices associated with Sonic hedgehog (Shh) signalling and ciliopathies. Here, using mammalian cells and zebrafish, we uncover ciliary functions for Rab35, a multitasking G protein with endocytic recycling, actin remodelling and cytokinesis roles. Rab35 loss via siRNAs, morpholinos or knockout reduces cilium length in mammalian cells and the zebrafish left-right organiser (Kupffer's vesicle) and causes motile cilia-associated left-right asymmetry defects. Consistent with these observations, GFP-Rab35 localises to cilia, as do GEF (DENND1B) and GAP (TBC1D10A) Rab35 regulators, which also regulate ciliary length and Rab35 ciliary localisation. Mammalian Rab35 also controls the ciliary membrane levels of Shh signalling regulators, promoting ciliary targeting of Smoothened, limiting ciliary accumulation of Arl13b and the inositol polyphosphate 5-phosphatase (INPP5E). Rab35 additionally regulates ciliary PI(4,5)P2 levels and interacts with Arl13b. Together, our findings demonstrate roles for Rab35 in regulating cilium length, function and membrane composition and implicate Rab35 in pathways controlling the ciliary levels of Shh signal regulators.


Fine-root respiration in a loblolly pine and sweetgum forest growing in elevated CO2.
• The loss of carbon below-ground through respiration of fine roots may be modified by global change. Here we tested the hypothesis that a reduction in N concentration of tree fine-roots grown in an elevated atmospheric CO2 concentration would reduce maintenance respiration and that more energy would be used for root growth and N uptake. We partitioned total fine-root respiration (RT ) between maintenance (RM ), growth (RG ), and N uptake respiration (RN ) for loblolly pine (Pinus taeda) and sweetgum (Liquidambar styraciflua) forests exposed to elevated CO2 . • A substantial increase in fine-root production contributed to a 151% increase in RG for loblolly pine in elevated CO2 . Root specific RM for pine was 24% lower under elevated CO2 but when extrapolated to the entire forest, no treatment effect could be detected. • R G (< 10%) and RN (< 3%) were small components of RM in both forests. Maintenance respiration was the vast majority of RT , and contributed 92% and 86% of these totals at the pine and sweetgum forests, respectively. • The hypothesis was rejected because the majority of fine-root respiration was used for maintenance and was not reduced by changes in root N concentration in elevated CO2 . Because of its large contribution to RT and total soil CO2 efflux, changes in RM caused by warming may greatly alter carbon losses from forests to the atmosphere.


Stability and mismatch discrimination of locked nucleic acid-DNA duplexes.
Locked nucleic acids (LNA; symbols of bases, +A, +C, +G, and +T) are introduced into chemically synthesized oligonucleotides to increase duplex stability and specificity. To understand these effects, we have determined thermodynamic parameters of consecutive LNA nucleotides. We present guidelines for the design of LNA oligonucleotides and introduce free online software that predicts the stability of any LNA duplex oligomer. Thermodynamic analysis shows that the single strand-duplex transition is characterized by a favorable enthalpic change and by an unfavorable loss of entropy. A single LNA modification confines the local conformation of nucleotides, causing a smaller, less unfavorable entropic loss when the single strand is restricted to the rigid duplex structure. Additional LNAs adjacent to the initial modification appear to enhance stacking and H-bonding interactions because they increase the enthalpic contributions to duplex stabilization. New nearest-neighbor parameters correctly forecast the positive and negative effects of LNAs on mismatch discrimination. Specificity is enhanced in a majority of sequences and is dependent on mismatch type and adjacent base pairs; the largest discriminatory boost occurs for the central +C·C mismatch within the +T+C+C sequence and the +A·G mismatch within the +T+A+G sequence. LNAs do not affect specificity in some sequences and even impair it for many +G·T and +C·A mismatches. The level of mismatch discrimination decreases the most for the central +G·T mismatch within the +G+G+C sequence and the +C·A mismatch within the +G+C+G sequence. We hypothesize that these discrimination changes are not unique features of LNAs but originate from the shift of the duplex conformation from B-form to A-form.


Systemic and immunotoxicity of silver nanoparticles in an intravenous 28 days repeated dose toxicity study in rats.
Because of its antibacterial activity nanosilver is one of the most commonly used nanomaterials. It is increasingly used in a variety of both medical and consumer products resulting in an increase in human exposure. However, the knowledge on the systemic toxicity of nanosilver is relatively limited. To determine the potential systemic toxicity of silver nanoparticles (Ag-NP) with different sizes (20 nm and 100 nm) a 28-days repeated dose toxicity study was performed in rats using intravenous administration. The toxic effect of the 20 nm Ag-NP was performed using the bench mark dose (BMD) approach. Treatment with a maximum dose of 6 mg/kg body weight was well tolerated by the animals. However, both for 20 nm and 100 nm Ag-NP growth retardation was observed during the treatment. A severe increase in spleen size and weight was present which was due to an increased cell number. Both T and B cell populations showed an increase in absolute cell number, whereas the relative cell numbers remained constant. At histopathological evaluation brown and black pigment indicating Ag-NP accumulation was noted in spleen, liver, and lymph nodes. Ag-NP was also detected incidentally in other organs. Clinical chemistry indicated liver damage (increased alkaline phosphatase, alanine transaminase, and aspartate transaminase) that could not be confirmed by histopathology. Hematology showed a decrease in several red blood cell parameters. The most striking toxic effect was the almost complete suppression of the natural killer (NK) cell activity in the spleen at high doses. Other immune parameters affected were: decreased interferon-γ and interleukin (IL)-10 production by concanavalin-A stimulated spleen cells, increased IL-1β and decreased IL-6, IL-10 and TNF-α production by lipopolysaccharide stimulated spleen cells, increase in serum IgM and IgE, and increase in blood neutrophilic granulocytes. For the spleen weight a critical effect dose of 0.37 mg/kg body weight (b.w.) could be established. The lowest critical effect dose (CED) for a 5% change compared to control animals was observed for thymus weight (CED05 0.01 mg/kg b.w.) and for functional immune parameters, i.e. decrease in NK cell activity (CED05 0.06 mg/kg b.w.) and LPS stimulation of spleen cells (CED05 0.04 mg/kg b.w.). These results show that for nanosilver the most sensitive parameters for potential adverse responses were effects on the immune system.


Little evidence for association between the TGFBR1*6A variant and colorectal cancer: a family-based association study on non-syndromic family members from Australia and Spain.
Genome-wide linkage studies have identified the 9q22 chromosomal region as linked with colorectal cancer (CRC) predisposition. A candidate gene in this region is transforming growth factor β receptor 1 (TGFBR1). Investigation of TGFBR1 has focused on the common genetic variant rs11466445, a short exonic deletion of nine base pairs which results in truncation of a stretch of nine alanine residues to six alanine residues in the gene product. While the six alanine (*6A) allele has been reported to be associated with increased risk of CRC in some population based study groups this association remains the subject of robust debate. To date, reports have been limited to population-based case-control association studies, or case-control studies of CRC families selecting one affected individual per family. No study has yet taken advantage of all the genetic information provided by multiplex CRC families. We have tested for an association between rs11466445 and risk of CRC using several family-based statistical tests in a new study group comprising members of non-syndromic high risk CRC families sourced from three familial cancer centres, two in Australia and one in Spain. We report a finding of a nominally significant result using the pedigree-based association test approach (PBAT; p = 0.028), while other family-based tests were non-significant, but with a p-value <; 0.10 in each instance. These other tests included the Generalised Disequilibrium Test (GDT; p = 0.085), parent of origin GDT Generalised Disequilibrium Test (GDT-PO; p = 0.081) and empirical Family-Based Association Test (FBAT; p = 0.096, additive model). Related-person case-control testing using the "More Powerful" Quasi-Likelihood Score Test did not provide any evidence for association (MQLS; p = 0.41). After conservatively taking into account considerations for multiple hypothesis testing, we find little evidence for an association between the TGFBR1*6A allele and CRC risk in these families. The weak support for an increase in risk in CRC predisposed families is in agreement with recent meta-analyses of case-control studies, which estimate only a modest increase in sporadic CRC risk among 6*A allele carriers.


Development and Evaluation of Self-Management and Task-Oriented Approach to Rehabilitation Training (START) in the Home: Case Report.
The incidence of stroke and subsequent level of disability will increase, as age is the greatest risk factor for stroke and the world's population is aging. Hospital admissions are too brief for patients to regain necessary function. Research to examine therapy delivered within the home environment has the potential to expedite relearning of function and reduce health care expenditures. This case report describes the use of the knowledge-to-action cycle (KTA) to develop and evaluate an evidence-based approach for rehabilitation in the home that incorporates self-management and task-oriented functional training (TOFT) for people with stroke. The KTA cycle was used to guide adaptation of evidence from self-management and TOFT into an approach titled START (Self-Management and Task-Oriented Approach to Rehabilitation Training). Three stakeholder symposiums identified barriers and supports to implementation. Clinical practice leaders were engaged as partners in the development of the intervention. An online learning management system housed the resources to support therapist training. Therapist focus groups were conducted and stroke outcomes were used to assess patient response. Eight therapists completed 4 workshops and applied the home intervention in 12 people with stroke. A mentoring process for therapists included feedback from peers and experts after viewing treatment videos. Therapist response was determined from the focus groups; patient response was measured by standardized assessments. The therapists noted that the intervention was easier to implement with patients who were motivated and had minimal cognitive impairment. The KTA cycle provided a structure for the development of this evidence-based rehabilitation intervention, which was feasible to implement in the home. Further evaluation needs to be undertaken to assess the effectiveness of START.


Examiner training: A study of examiners making sense of norm-referenced feedback.
Purpose: Examiner training has an inconsistent impact on subsequent performance. To understand this variation, we explored how examiners think about changing the way they assess. Method: We provided comparative data to 17 experienced examiners about their assessments, captured their sense-making processes using a modified think-aloud protocol, and identified patterns by inductive thematic analysis. Results: We observed five sense-making processes: (1) testing personal relevance (2) interpretation (3) attribution (4) considering the need for change, and (5) considering the nature of change. Three observed meta-themes describe the manner of examiners' thinking: Guarded curiosity - where examiners expressed curiosity over how their judgments compared with others', but they also expressed guardedness about the relevance of the comparisons; Dysfunctional assimilation - where examiners' interpretation and attribution exhibited cognitive anchoring, personalization, and affective bias; Moderated conservatism - where examiners expressed openness to change, but also loyalty to their judgment-framing values and aphorisms. Conclusions: Our examiners engaged in complex processes as they considered changing their assessments. The 'stabilising' mechanisms some used resembled learners assimilating educational feedback. If these are typical examiner responses, they may well explain the variable impact of examiner training, and have significant implications for the pursuit of meaningful and defensible judgment-based assessment.


Unravelling the Bacterial Vaginosis-Associated Biofilm: A Multiplex Gardnerella vaginalis and Atopobium vaginae Fluorescence In Situ Hybridization Assay Using Peptide Nucleic Acid Probes.
Bacterial vaginosis (BV), a condition defined by increased vaginal discharge without significant inflammation, is characterized by a change in the bacterial composition of the vagina. Lactobacillus spp., associated with a healthy vaginal microbiome, are outnumbered by BV-associated organisms. These bacteria could form a polymicrobial biofilm which allows them to persist in spite of antibiotic treatment. In this study, we examined the presence of Gardnerella vaginalis and Atopobium vaginae in vaginal biofilms using Peptide Nucleic Acid (PNA) probes targeting these bacteria. For this purpose, we developed three new PNA probes for A. vaginae. The most specific A. vaginae probe, AtoITM1, was selected and then used in an assay with two existing probes, Gard162 and BacUni-1, to evaluate multiplex FISH on clinical samples. Using quantitative polymerase chain reaction (qPCR) as the gold standard, we demonstrated a sensitivity of 66.7% (95% confidence interval: 54.5% - 77.1%) and a specificity of 89.4% (95% confidence interval: 76.1% - 96%) of the new AtoITM1 probe. FISH enabled us to show the presence of a polymicrobial biofilm in bacterial vaginosis, in which Atopobium vaginae is part of a Gardnerella vaginalis-dominated biofilm. We showed that the presence of this biofilm is associated with high bacterial loads of A. vaginae and G. vaginalis.


Variants at the APOA5 locus, association with carotid atherosclerosis, and modification by obesity: the Framingham Study.
Genetic variation at the apolipoprotein A5 gene (APOA5) is associated with increased triglyceride concentrations, a risk factor for atherosclerosis. Carotid intimal medial thickness (IMT) is a surrogate measure of atherosclerosis burden. We sought to determine the association of common APOA5 genetic variants with carotid IMT and stenosis. A total of 2,273 Framingham Offspring Study participants underwent carotid ultrasound and had data on at least one of the five APOA5 variants (-1131T>C, -3A>G, 56C>G, IVS3+476G >A, and 1259T>C). Although none of the individual variants was significantly associated with carotid measures, the haplotype defined by the presence of the rare allele of the 56C>G variant was associated with a higher common carotid artery (CCA) IMT compared with the wild-type haplotype (0.75 vs. 0.73 mm; P < 0.05). The rare allele of each of the -1131T >C, -3A>G, IVS3+476G>A, and 1259T>C variants and the haplotype defined by the presence of the rare alleles in these four variants were each significantly associated with CCA IMT in obese participants. These associations remained significant even after adjustment for triglycerides. APOA5 variants were associated with CCA IMT, particularly in obese participants. The mechanism of these associations and the effect modification by obesity are independent of fasting triglyceride levels.


Neural Correlates of Divided Attention in Natural Scenes.
Individuals are able to split attention between separate locations, but divided spatial attention incurs the additional requirement of monitoring multiple streams of information. Here, we investigated divided attention using photos of natural scenes, where the rapid categorization of familiar objects and prior knowledge about the likely positions of objects in the real world might affect the interplay between these spatial and nonspatial factors. Sixteen participants underwent fMRI during an object detection task. They were presented with scenes containing either a person or a car, located on the left or right side of the photo. Participants monitored either one or both object categories, in one or both visual hemifields. First, we investigated the interplay between spatial and nonspatial attention by comparing conditions of divided attention between categories and/or locations. We then assessed the contribution of top-down processes versus stimulus-driven signals by separately testing the effects of divided attention in target and nontarget trials. The results revealed activation of a bilateral frontoparietal network when dividing attention between the two object categories versus attending to a single category but no main effect of dividing attention between spatial locations. Within this network, the left dorsal premotor cortex and the left intraparietal sulcus were found to combine task- and stimulus-related signals. These regions showed maximal activation when participants monitored two categories at spatially separate locations and the scene included a nontarget object. We conclude that the dorsal frontoparietal cortex integrates top-down and bottom-up signals in the presence of distractors during divided attention in real-world scenes.


Dynamics of confined water and its interplay with alkali cations in sodium aluminosilicate hydrate gel: insights from reactive force field molecular dynamics.
This paper presents the dynamics of confined water and its interplay with alkali cations in disordered sodium aluminosilicate hydrate (N-A-S-H) gel using reactive force field molecular dynamics. N-A-S-H gel is the primary binding phase in geopolymers formed via alkaline activation of fly ash. Despite attractive mechanical properties, geopolymers suffer from durability issues, particularly the alkali leaching problem which has motivated this study. Here, the dynamics of confined water and the mobility of alkali cations in N-A-S-H is evaluated by obtaining the evolution of mean squared displacements and Van Hove correlation function. To evaluate the influence of the composition of N-A-S-H on the water dynamics and diffusion of alkali cations, atomistic structures of N-A-S-H with Si/Al ratio ranging from 1 to 3 are constructed. It is observed that the diffusion of confined water and sodium is significantly influenced by the Si/Al ratio. The confined water molecules in N-A-S-H exhibit a multistage dynamic behavior where they can be classified as mobile and immobile water molecules. While the mobility of water molecules gets progressively restricted with an increase in Si/Al ratio, the diffusion coefficient of sodium also decreases as the Si/Al ratio increases. The diffusion coefficient of water molecules in the N-A-S-H structure exhibits a lower value than those of the calcium-silicate-hydrate (C-S-H) structure. This is mainly due to the random disordered structure of N-A-S-H as compared to the layered C-S-H structure. To further evaluate the influence of water content in N-A-S-H, atomistic structures of N-A-S-H with water contents ranging from 5-20% are constructed. Qn distribution of the structures indicates significant depolymerization of N-A-S-H structure with increasing water content. Increased conversion of Si-O-Na network to Si-O-H and Na-OH components with an increase in water content helps explain the alkali-leaching issue in fly ash-based geopolymers observed macroscopically. Overall, the results in this study can be used as a starting point towards multiscale simulation-based design and development of durable geopolymers.


Promoter hypermethylation in circulating blood cells identifies prostate cancer progression.
Promoter hypermethylation of circulating cell DNA has been advocated as a diagnostic marker for prostate cancer, but its prognostic use is currently unclear. To assess this role, we compared hypermethylation of circulating cell DNA from prostate cancer patients with (Group 1, n = 20) and without (Group 2, n = 22) disease progression and age-matched controls (benign prostatic hyperplasia, Group 3, n = 22). We measured hypermethylation of 10 gene promoters in 2 sequential venous samples, obtained at diagnosis and during disease progression (median time, 15 months later). Matched time samples were obtained in the nonprogressing patients. We found that more hypermethylation was detected in the diagnostic sample from the patients with cancer than in controls for GSTP1, RASSF1 alpha, APC and RAR beta (p < 0.0001). Patients undergoing disease progression had a significant increase in methylation levels of these 4 genes when compared to the other patients (p < 0.001). Patients at risk of disease progression have higher detectable concentrations of circulating cell hypermethylation, than those without progression. The extent of this hypermethylation increases during disease progression and can be used to identify the extent and duration of treatment response in prostate cancer.


Incidence, Long-Term Visual Outcomes, and Mortality in Retinopathy of Prematurity in Korea: A Nationwide Population-Based Study.
To evaluate the incidence, visual prognosis, and mortality in retinopathy of prematurity (ROP) in Korea. We used the National Health Insurance and the Korean Disability Registry database, which covers the entire newborn population in 2006 to 2014 and includes information on all newborns diagnosed with ROP until 2016. Using these databases, we evaluated the incidence, rate of visual impairment (VI), and mortality in patients with ROP according to the birth weight categories and treatment modalities. The ROP incidence per 1000 newborns was 1.99, which broke down into 317.14 in the very low birth weight (VLBW) less than 1500 g population, 25.45 in the 1500 to 2499 g population, and 0.29 in the 2500 g or greater population. When assessed at age 10, the VI rate was 2.2 per 100 person-years, which was highest at 4.5 per 100 person-years in the VLBW population compared with the population in other birth weight categories. Among treated cases, the proportion of VI in patients undergoing laser photocoagulation or cryotherapy was 1.6% (42/2595), which was lower than the 2.9% (2/68) of patients treated with anti-vascular endothelial growth factor injection, and 32.2% (82/255) of patients undergoing vitrectomy or scleral buckling. The mortality rate was 4.8 per 1000 person-years, which was highest in the VLBW population, but similar across treatment modalities. The ROP incidence in Korea was approximately 1 in 500 among all newborns, and 1 in 3 in the VLBW population. As the first nationwide population-based study of long-term visual prognosis in ROP, we report the higher VI rate in ROP than previously determined in other studies. Differences in visual outcomes and comparable mortality risks between treatment modalities require further verification.


Combination of arginine deprivation with TRAIL treatment as a targeted-therapy for mesothelioma.
In the present study we present data to show that certain tumor cells including malignant pleural mesothelioma (MPM) cells do not express argininosuccinate synthetase (ASS), and thus are unable to synthesize arginine from citrulline. Exposure of these ASS-negative cells to the arginine degrading enzyme, arginine deiminase (ADI-PEG20), for 72 h results in significant increases in cleaved caspase-3. Importantly, this apoptotic signal is further strengthened by the addition of TNF-related apoptosis-inducing ligand (TRAIL). Using flow cytometry, we showed that the combination treatment (ADI-PEG20 at 50 ng/ml and TRAIL at 10 ng/ml) for 24 h resulted in profound cell death with 67% of cells positive for caspase-3 activity, while ADI-PEG20 alone or TRAIL alone resulted in only 10-15% cell death. This positive amplification loop is mediated through the cleavage of proapototic protein "BID". Our work represents a new strategy for treating patients with malignant pleural mesothelioma using targeted molecular therapeutics based on selected tumor markers, thus avoiding the use of potentially cytotoxic chemotherapy.


Synchronized action of synaptically coupled chaotic model neurons.
Experimental observations of the intracellular recorded electrical activity in individual neurons show that the temporal behavior is often chaotic. We discuss both our own observations on a cell from the stomatogastric central pattern generator of lobster and earlier observations in other cells. In this paper we work with models with chaotic neurons, building on models by Hindmarsh and Rose for bursting, spiking activity in neurons. The key feature of these simplified models of neurons is the presence of coupled slow and fast subsystems. We analyze the model neurons using the same tools employed in the analysis of our experimental data. We couple two model neurons both electrotonically and electrochemically in inhibitory and excitatory fashions. In each of these cases, we demonstrate that the model neurons can synchronize in phase and out of phase depending on the strength of the coupling. For normal synaptic coupling, we have a time delay between the action of one neuron and the response of the other. We also analyze how the synchronization depends on this delay. A rich spectrum of synchronized behaviors is possible for electrically coupled neurons and for inhibitory coupling between neurons. In synchronous neurons one typically sees chaotic motion of the coupled neurons. Excitatory coupling produces essentially periodic voltage trajectories, which are also synchronized. We display and discuss these synchronized behaviors using two "distance" measures of the synchronization.


Tensile properties of resin-infiltrated demineralized human dentin.
The ability of adhesive resins to restore the physical properties of demineralized dentin has not been well-documented. The unfilled resins that are used for adhesion have relatively low moduli of elasticity and limited ability to increase dentin stiffness, although they may increase the ultimate tensile strength of dentin. This study tested the hypothesis that resin infiltration of demineralized dentin can restore its tensile properties to those of mineralized dentin. Small (ca. 0.5 mm thick x 0.5 mm wide) specimens of demineralized human dentin were infiltrated with one of five different dentin bonding resins over many hours, to determine how these resins altered the tensile properties of dentin. Tensile stress and strain were measured in these and control (mineralized and demineralized) specimens until their ultimate failure. The results indicate that some adhesive resins, after infiltrating demineralized dentin, can restore and even exceed the ultimate tensile strength of mineralized dentin. These resins increased the modulus of elasticity of resin-infiltrated dentin to values equal to or greater than those of the resins but far below those of mineralized dentin. Although the conditions in this experiment were far removed from the manufacturer's recommendations or clinical practice, the results support the potential of resin infiltration for reinforcing dentin.


Evaluation of BG-Sentinel Trap as a Management Tool to Reduce Aedes albopictus Nuisance in an Urban Environment in Italy.
Since its introduction and establishment in Italy during the early 1990s, the Asian tiger mosquito, Aedes albopictus, has spread over large parts of Italy and other Mediterranean countries. Aedes albopictus is both a nuisance and a competent vector for various arthropod-borne pathogens. Although efficient traps for Ae. albopictus exist and are used for population monitoring, their use as a control tool has not yet been studied. We evaluated Biogents BG-Sentinel mosquito traps, used with the BG Lure, as control tools in northern Italy. The trial was performed as a controlled experiment in which 3 intervention sites, equipped with 7 or 8 BG-Sentinel traps each, were matched with 3 comparable control sites. Trap density ranged from 1 trap per 150 m² to 1 per 350 m². Mosquito populations were monitored at both the intervention and control sites with weekly human landing collections (HLC) and ovitraps. Between 64% and 87% fewer Ae. albopictus individuals were collected by HLC at the intervention sites with the BG-Sentinel mosquito traps, as compared to the untreated control sites. These results indicate that the sustained use and proper placement of efficient mosquito traps can significantly reduce Ae. albopictus biting pressure.


'Getting back to normal' or 'a new type of normal'? A qualitative study of patients' responses to the existential threat of cancer.
Existential concerns about cancer have been studied extensively in palliative care but less so in curative settings. The present report aims to describe ways in which patients viewed the continuity or discontinuity of their identity in the face of the mortal threat of cancer. Twenty-eight patients with breast, prostate or lung cancer attending pre-treatment, treatment or follow-up appointments were interviewed about their emotional experiences following diagnosis. Qualitative analysis followed an inductive, constant comparative approach. Patients spoke of 'getting back to normal', but presented two distinct accounts of 'normality'. Some, particularly those only recently diagnosed, maintained continuity to past identity by upholding previous routines, emphasising resilience and minimising the impact of cancer. Others talked of a new 'normality' discontinuous with their past. Most accounts, however, evidenced elements of continuity and discontinuity, often in ostensibly contradictory ways. We suggest that holding contradictory perspectives simultaneously characterises an intermediate stage of adjustment for some patients: between reliance on continuity with the past in the aftermath of diagnosis and, later, a sense of being a new person, changed by cancer. Practitioners should appreciate that patients' wishes for 'normality' can signify very different responses to cancer, and that holding such contradictory orientations is functional, not aberrant.


Hepatoprotective and antioxidant activities of grapeseeds against ethanol-induced oxidative stress in rats.
The present study was carried out to evaluate the hepatoprotective effect and antioxidant role of grape (Vitis vinifera L.) seeds (GS) against ethanol-induced oxidative stress. The hepatoprotective and antioxidant roles of the GS supplementation feed against ethanol-induced oxidative stress were evaluated by measuring liver damage serum marker enzymes, aspartate aminotransferase, alanine aminotransferase, γ-glutamyl transpeptidase and lactate dehydrogenase, antioxidant defence system such as GSH, glutathione reductase, superoxide dismutase, glutathione S-transferase and glutathione peroxidase and malondialdehyde (MDA) content in various tissues of rats. Rats were divided into four experimental groups: I (control), II (20 % ethanol), III (15 % GS) and IV (20 % ethanol+15 % GS). According to the results, the level of serum marker enzymes was significantly increased in group II as compared to that of group I, but decreased in group IV as compared to that of group II. Also, administration of GS-supplemented food restored the ethanol-induced MDA, which was increased near the control level. The results indicated that GS could be as important as diet-derived antioxidants in preventing oxidative damage in the tissues by reducing the lipid oxidation or inhibiting the production of ethanol-induced free radicals in rats.


HBV, HCV and HIV seroprevalence among blood donors in Istanbul, Turkey: how effective are the changes in the national blood transfusion policies?
The national blood transfusion policies have been changed significantly in recent years in Turkey. The purpose of this study was to determine the prevalence of HBV, HCV, and HIV in blood donors at the Red Crescent Center in Istanbul and to evaluate the effect of changes in the national blood transfusion policies on the prevalence of these infections. The screening results of 72695 blood donations at the Red Crescent Center in Istanbul between January and December 2007 were evaluated retrospectively. HBsAg, anti-HCV, and anti-HIV-1/2 were screened by microparticle enzyme immunoassay (MEIA) method. Samples found to be positive for anti-HIV 1/2 and anti-HCV were confirmed by Inno-Lia HCV Ab III and Inno-Lia HIV I/II Score, respectively. The seropositivity rates for HBsAg, anti-HCV, and anti-HIV-1/2 were determined as 1.76%, 0.07%, and 0.008%, respectively. Compared to the previously published data from Red Crescent Centers in Turkey, it was found that HBV and HCV seroprevalances decreased and HIV seroprevalance increased in recent years. In conclusion, we believe that the drop in HBV and HCV prevalence rates are likely multifactorial and may have resulted from more diligent donor questioning upon screening, a higher level of public awareness on viral hepatitis as well as the expansion of HBV vaccination coverage in Turkey. Another factor to contribute to the decreased prevalence of HCV stems from the use of more sensitive confirmation testing on all reactive results, thereby eliminating a fair amount of false positive cases. Despite similar transmission routes, the increase in HIV prevalence in contrast to HBV and HCV may be linked to the increase in AIDS cases in Turkey in recent years.


A FRET analysis to unravel the role of cholesterol in Rac1 and PI 3-kinase activation in the InlB/Met signalling pathway.
The signalling pathway for the hepatocyte growth factor receptor, Met/HGF-R, is hijacked by the bacterial surface protein InlB to induce Listeria monocytogenes entry into non-phagocytic cells. We previously showed that Listeria invades host cells by interacting with specialized microdomains of the host plasma membrane called lipid rafts. In this study, we analysed in living cells signalling events that are crucial for Listeria entry using a fluorescence resonance energy transfer-based microscopic method. Phosphoinositide (PI) 3-kinase activity and Rac1 signalling induced by Listeria interacting with epithelial cells were monitored as well as signalling induced by soluble InlB and the Met natural ligand HGF. We found that InlB and HGF induced similar kinetics of PI 3-kinase and Rac1 activation. PI 3-kinase activation was upstream and independent of Rac1 activation. Cholesterol-depletion experiments were performed to address the role of lipid rafts in Met signalling. The amount of 3'-phosphoinositides produced by PI 3-kinase was not affected by cholesterol depletion, while their membrane dynamic was cholesterol-dependent. Rac1 activation, downstream from PI 3-kinase, was cholesterol-dependent suggesting that the spatial distribution of 3'-phosphoinositides within membrane microdomains is critical for Rac1 activation and consequently for F-actin assembly at bacterial entry site.


A note on the analysis of two-stage task results: How changes in task structure affect what model-free and model-based strategies predict about the effects of reward and transition on the stay probability.
Many studies that aim to detect model-free and model-based influences on behavior employ two-stage behavioral tasks of the type pioneered by Daw and colleagues in 2011. Such studies commonly modify existing two-stage decision paradigms in order to better address a given hypothesis, which is an important means of scientific progress. It is, however, critical to fully appreciate the impact of any modified or novel experimental design features on the expected results. Here, we use two concrete examples to demonstrate that relatively small changes in the two-stage task design can substantially change the pattern of actions taken by model-free and model-based agents as a function of the reward outcomes and transitions on previous trials. In the first, we show that, under specific conditions, purely model-free agents will produce the reward by transition interactions typically thought to characterize model-based behavior on a two-stage task. The second example shows that model-based agents' behavior is driven by a main effect of transition-type in addition to the canonical reward by transition interaction whenever the reward probabilities of the final states do not sum to one. Together, these examples emphasize the task-dependence of model-free and model-based behavior and highlight the benefits of using computer simulations to determine what pattern of results to expect from both model-free and model-based agents performing a given two-stage decision task in order to design choice paradigms and analysis strategies best suited to the current question.


Effects of acrivastine, loratadine and cetirizine on histamine-induced wheal and flare responses.
It is accepted that studies evaluating histamine-induced wheal and flare reactions in the skin represent a simple and reliable method for demonstrating pharmacodynamic activity and pharmacokinetics of the H1-receptor antagonists. In this study, the effects of single oral doses of acrivastine (8 mg), loratadine (10 mg) and cetirizine (10 mg) on the histamine-induced wheal and flare reactions were compared in 60 healthy volunteers. The wheal and flare responses were produced by prick test using 1% histamine solution. Measurements were performed before the ingestion of antihistamines (baseline values) and afterwards at 15, 30, 90, 240, 360 min and 24 h. The values obtained for each antihistamine were compared with each other and with baseline values. Cetirizine was found to be superior to acrivastine and loratadine for the suppression of wheal and flare responses at 240, 360 min and 24 h (P < 0.05) and acrivastine was superior to the other two antihistamines for the suppression of flare response at 30 min (P < 0.05). Our results indicate that a single dose of cetirizine provides a more effective and long acting suppression on wheal and flare reactions in urticaria when compared to acrivastine and loratadine.