The role of vitamin D on circulating memory T cells in children: The Generation R study.
Previous studies have demonstrated that vitamin D affects T-cell function and maturation via the vitamin D receptor. However, no studies in children have been performed on this topic. Because most of the T-cell memory is formed in the first 5 years of life, we aimed to determine the association between serum 25-hydroxyvitamin D (25(OH)D) levels and numbers of circulatory naive, central memory (Tcm), and effector memory (Tem) T lymphocytes in a large population of healthy children. Among 3189 children participating in a population-based prospective cohort, we measured 25(OH)D levels and performed detailed immunophenotyping of naive and memory T lymphocytes at a median age of 6.0 years (95% range 5.7-7.9). Detailed lymphocyte subsets were available in 986 children. Multivariable linear regression analyses were performed to determine the association between 25(OH)D and the maturation of T lymphocytes in children adjusted for cord blood 25(OH)D levels, herpes seropositivity, sociodemographic and lifestyle confounders. Furthermore, multivariable logistic regression analyses were performed to determine associations between 25(OH)D and childhood infections. Higher 25(OH)D levels were associated with higher numbers of Tem lymphocytes. Every 10 nmol/L higher 25(OH)D was associated with 2.20% (95% CI 0.54-3.89; P=.009) higher CD4TemRA, 1.50% (95% CI 0.38-2.62; P=.008) higher CD4TemRO, and 1.82% (95% CI 0.11-3.56; P=.037) higher CD8TemRA cell numbers. Generally, stronger associations were observed among boys. 25(OH)D levels were not significantly associated with naive, Tcm cell numbers, herpes seropositivity, or URTIs. Our results suggest that vitamin D enhances cellular immunity in young children.


Digital processing with a three-state molecular switch.
Certain molecular switches respond to input stimulations producing detectable outputs. The interplay of these signals can be exploited to reproduce basic logic operations at the molecular level. The transition from simple logic gates to complex digital circuits requires the design of chemical systems able to process multiple inputs and outputs. We have identified a three-state molecular switch that responds to one chemical and two optical inputs producing two optical outputs. We have encoded binary digits in its inputs and outputs applying positive logic conventions and demonstrated that this chemical system converts three-digit input strings into two-digit output strings. The logic function executed by the three-state molecular switch is equivalent to that of a combinational logic circuit integrating two AND, two NOT, and one OR gate. The three states of the molecular switch are a colorless spiropyran, a purple trans-merocyanine, and its yellow-green protonated form. We have elucidated their structures by X-ray crystallography, (1)H NMR spectroscopy, COSY and NOE experiments, as well as density functional calculations. The three input stimulations controlling the interconversion of the three states of the molecular switch are ultraviolet light, visible light, and H(+). The two outputs are the absorption bands in the visible region of the two colored states of the molecular switch. We have monitored the switching processes and quantified the associated thermodynamic and kinetic parameters with the aid of (1)H NMR and visible absorption spectroscopies.


Hot flushes among aging women: A 4-year follow-up study to a randomised controlled exercise trial.
The aim of this follow-up study was to explore the long-term effects of a 6-month trial of exercise training on hot flushes. The follow-up was 4 years after the exercise intervention ended. A cohort study after a randomised controlled trial. Ninety-five of the 159 randomised women (60%) participated in anthropometric measurements and performed a 2-km walk test. Participants completed a questionnaire and kept a one-week diary on physical activity, menopause symptoms and sleep quality. The frequency of 24-h hot flushes was multiplied by severity and the total sum for one week was defined as the Hot Flush Score (HFScore). Multilevel mixed regression models were analysed to compare the exercise and control groups. Hot Flush Score (HFScore) as assessed with the one-week symptom diary. The women in the exercise group had a higher probability of improved HFScore, i.e. a decrease in HFScore points, adjusted for hormone therapy (OR 0.95; 95% CI 0.90-1.00) than women in the control group at the 4-year follow-up. After additional adjustment for sleep quality, the result approached statistical significance at HFScore≥13 with women in the exercise group. Women who had the least amount of hot flushes, HFScore<13, benefited most from exercise during the 4-year follow-up when compared with women in the control group. Women in the exercise group had positive effects on their HFScore 4 years after a 6-month exercise intervention.


Tachistoscopic illumination and masking of real scenes.
Tachistoscopic presentation of scenes has been valuable for studying the emerging properties of visual scene representations. The spatial aspects of this work have generally been focused on the conceptual locations (e.g., next to the refrigerator) and directional locations of objects in 2-D arrays and/or images. Less is known about how the perceived egocentric distance of objects develops. Here we describe a novel system for presenting brief glimpses of a real-world environment, followed by a mask. The system includes projectors with mechanical shutters for projecting the fixation and masking images, a set of LED floodlights for illuminating the environment, and computer-controlled electronics to set the timing and initiate the process. Because a real environment is used, most visual distance and depth cues can be manipulated using traditional methods. The system is inexpensive, robust, and its components are readily available in the marketplace. This article describes the system and the timing characteristics of each component. We verified the system's ability to control exposure to time scales as low as a few milliseconds.


Harmful freshwater algal blooms, with an emphasis on cyanobacteria.
Suspended algae, or phytoplankton, are the prime source of organic matter supporting food webs in freshwater ecosystems. Phytoplankton productivity is reliant on adequate nutrient supplies; however, increasing rates of nutrient supply, much of it manmade, fuels accelerating primary production or eutrophication. An obvious and problematic symptom of eutrophication is rapid growth and accumulations of phytoplankton, leading to discoloration of affected waters. These events are termed blooms. Blooms are a prime agent of water quality deterioration, including foul odors and tastes, deoxygenation of bottom waters (hypoxia and anoxia), toxicity, fish kills, and food web alterations. Toxins produced by blooms can adversely affect animal (including human) health in waters used for recreational and drinking purposes. Numerous freshwater genera within the diverse phyla comprising the phytoplankton are capable of forming blooms; however, the blue-green algae (or cyanobacteria) are the most notorious bloom formers. This is especially true for harmful toxic, surface-dwelling, scum-forming genera (e.g., Anabaena, Aphanizomenon, Nodularia, Microcystis) and some subsurface bloom-formers (Cylindrospermopsis, Oscillatoria) that are adept at exploiting nutrient-enriched conditions. They thrive in highly productive waters by being able to rapidly migrate between radiance-rich surface waters and nutrient-rich bottom waters. Furthermore, many harmful species are tolerant of extreme environmental conditions, including very high light levels, high temperatures, various degrees of desiccation, and periodic nutrient deprivation. Some of the most noxious cyanobacterial bloom genera (e.g., Anabaena, Aphanizomenon, Cylindrospermopsis, Nodularia) are capable of fixing atmospheric nitrogen (N2), enabling them to periodically dominate under nitrogen-limited conditions. Cyanobacteria produce a range of organic compounds, including those that are toxic to higher-ranked consumers, from zooplankton to further up the food chain. Both N2- and non-N2-fixing genera participate in mutualistic and symbiotic associations with microorganisms, higher plants, and animals. These associations appear to be of great benefit to their survival and periodic dominance. In this review, we address the ecological impacts and environmental controls of harmful blooms, with an emphasis on the ecology, physiology, and management of cyanobacterial bloom taxa. Combinations of physical, chemical, and biotic features of natural waters function in a synergistic fashion to determine the sensitivity of water bodies. In waters susceptible to blooms, human activities in water- and airsheds have been linked to the extent and magnitudes of blooms. Control and management of cyanobacterial and other phytoplankton blooms invariably includes nutrient input constraints, most often focused on nitrogen (N) and/or phosphorus (P). The types and amount of nutrient input constraints depend on hydrologic, climatic, geographic, and geologic factors, which interact with anthropogenic and natural nutrient input regimes. While single nutrient input constraints may be effective in some water bodies, dual N and P input reductions are usually required for effective long-term control and management of harmful blooms. In some systems where hydrologic manipulations (i.e., plentiful water supplies) are possible, reducing the water residence time by enhanced flushing and artificial mixing (in conjunction with nutrient input constraints) can be particularly effective alternatives. Implications of various management strategies, based on combined ecophysiological and environmental considerations, are discussed.


Efficiency of hospital cholera treatment in Ecuador.
This study analyzed the efficiency of cholera treatment in three hospitals representative of the Ecuadorian public health system in order to provide hospital directors and administrators and health service policy-makers with information to plan responses to future epidemics and to reduce the costs of cholera treatment in general. For the study, total and excess cholera treatment costs were calculated using hospital files and statistics and an in-hospital surveillance system of the cholera cases. The type and quantity of each input used for each treatment were analyzed, as well as the number of days hospitalized, according to the severity of the illness. With this process, excess costs were determined in relation to a "treatment norm" that would have been appropriate for each patient. The researchers found that 45% of the cholera treatment costs were excessive. The most important contributor was excess recurrent costs (90%), including extended hospital stays, disproportionate use of intravenous rehydration solutions, and unnecessary laboratory tests. Excess capital costs, from land, buildings, and hospital equipment, represented 10% of the total excess treatment costs. No significant relationship was found between treatment costs and the severity of the illness, nor between costs and a patient's age. A patient's sex appeared to be an important variable, with the cost of treating women being notably higher than for men. An inverse relationship was found between treatment costs and the complexity of the hospital. The researchers concluded there was an inefficient use of resources in the treatment of cholera in the three hospitals where the research was performed.


ALDH2 Gene: Its Effects on the Neuropsychological Functions in Patients with Opioid Use Disorder Undergoing Methadone Maintenance Treatment.
Patients with opioid use disorder (OUD) have impaired attention, inhibition control, and memory function. The aldehyde dehydrogenase 2(ALDH2) gene has been associated with OUD and ALDH2 gene polymorphisms may affect aldehyde metabolism and cognitive function in other substance use disorder. Therefore, we aimed to investigate whether ALDH2 genotypes have significant effects on neuropsychological functions in OUD patients undergoing methadone maintenance therapy (MMT). OUD patients undergoing MMT were investigated and followed-up for 12 weeks. ALDH2 gene polymorphisms were genotyped. Connors' Continuous Performance Test (CPT) and the Wechsler Memory Scale-Revised (WMS-R) were administered at baseline and after 12 weeks of MMT. Multivariate linear regressions and generalized estimating equations (GEEs) were used to examine the correlation between the ALDH2 genotypes and performance on the CPTs and WMS-R. We enrolled 86 patients at baseline; 61 patients completed the end-of-study assessments. The GEE analysis showed that, after the 12 weeks of MMT, OUD patients with the ALDH2 *1/*2+*2/*2 (ALDH2 inactive) genotypes had significantly higher commission error T-scores (p= 0.03), significantly lower hit reaction time T-scores (p= 0.04), and significantly lower WMS-R visual memory index scores (p= 0.03) than did patients with the ALDH2 1*/*1 (ALDH2 active) genotype. OUD patients with the ALDH2 inactive genotypes performed worse in cognitive domains of attention, impulse control, and memory than did those with the ALDH2 active genotype. We conclude that the ALDH2 gene is important in OUD and is associated with neuropsychological performance after MMT.


Identification and cloning of GOLDEN2-LIKE1 (GLK1), a transcription factor associated with chloroplast development in Brassica napus L.
Photosynthesis is the process by which dry matter accumulates, which affects rapeseed yield. In this study, we identified GOLDEN2-LIKE1 (GLK1), located on chromosome A07 and 59.2 kb away from the single nucleotide polymorphism marker SNP16353A07, which encodes a transcription factor associated with the rate of photosynthesis in leaves. We then identified 96 GLK1 family members from 53 species using a hidden Markov model (HMM) search and found 24 of these genes, which were derived from 17 Brassicaceae species. Phylogenetic analysis showed that 24 Brassicaceae proteins were classified into three subgroups, named the Brassica family, Adenium arabicum, and Arabidopsis. Using homologous cloning methods, we identified four BnaGLK1 copies; however, the coding sequences were shorter than the putative sequences from the reference genome, probably due to splicing errors among the reference genome sequence or different gene copies being present in the different B. napus lines. In addition, we found that BnaGLK1 genes were expressed at higher levels in leaves with more chloroplasts than were present in other leaves. Overexpression of BnaGLK1a resulted in darker leaves and siliques than observed in the control, suggesting that BnaGLK1 might promote chloroplast development to affect the rate of photosynthesis in leaves. These results will help to elucidate the mechanism of chloroplast biogenesis by GLK1 in B. napus.


Phase I dose-escalation study of plitidepsin in combination with sorafenib or gemcitabine in patients with refractory solid tumors or lymphomas.
This phase I trial evaluated the combination of the marine-derived cyclodepsipeptide plitidepsin (trade name Aplidin) with sorafenib or gemcitabine in advanced cancer and lymphoma patients. The study included two treatment arms: a sorafenib/plitidepsin (S/P) and a gemcitabine/plitidepsin (G/P) arm. In the S/P arm, patients were treated orally with sorafenib continuous dosing at two dose levels (DL1: 200 mg twice daily and DL2: 400 mg twice daily) combined with plitidepsin (1.8 mg/m, day 1, day 8, day 15, and, q4wk, intravenously). In the G/P arm, patients with solid tumors or lymphoma were treated at four different DLs with a combination of gemcitabine (DL1: 750 mg/m, DL2-DL4: 1000 mg/m) and plitidepsin (DL1-DL2: 1.8 mg/m; DL3: 2.4 mg/m; DL4: 3 mg/m). Both agents were administered intravenously on day 1, day 8, day 15, and, q4wk. Forty-four patients were evaluable for safety and toxicity. The safety of the combination of plitidepsin with sorafenib or gemcitabine was manageable. Most adverse events (AEs) were mild; no grade 4 treatment-related AEs were reported in any of the groups (except for one grade 4 thrombocytopenia in the gemcitabine arm). The most frequently reported study drug-related (or of unknown relationship) AEs were palmar-plantar erythrodysesthesia, erythema, nausea, vomiting, and fatigue in the S/P arm and nausea, fatigue, and vomiting in the G/P arm. In the S/P arm, one dose-limiting toxicity occurred in two out of six patients treated at the maximum dose tested (DL2): palmar-plantar erythrodysesthesia and grade 2 aspartate aminotransferase/alanine aminotransferase increase that resulted in omission of days 8 and 15 plitidepsin infusions. In the G/P arm, one dose-limiting toxicity occurred in two out of six patients at DL4: grade 2 alanine aminotransferase increase resulted in omission of days 8 and 15 plitidepsin infusions and grade 4 thrombocytopenia. The recommended dose for the combination of plitidepsin with sorafenib was not defined because of a sponsor decision (no expansion cohort to confirm) and for plitidepsin with gemcitabine, it was 2.4 mg/m plitidepsin with 1000 mg/m gemcitabine. In the S/P group, objective disease responses were not observed; however, disease stabilization (≥3months) was observed in four patients. In the gemcitabine group, two lymphoma patients showed an objective response (partial response and complete response) and nine patients showed disease stabilization (≥3months). Combining plitidepsin with gemcitabine and sorafenib is feasible for advanced cancer patients; some objective responses were observed in heavily pretreated lymphoma patients.


Right ventricular function in preterm and term neonates: reference values for right ventricle areas and fractional area of change.
Right ventricular (RV) fractional area of change (FAC) is a quantitative two-dimensional echocardiographic measurement of RV function. RV FAC expresses the percentage change in the RV chamber area between end-diastole (RV end-diastolic area [RVEDA]) to end-systole (RV end-systolic area [RVESA]). The objectives of this study were to determine the maturational (age- and weight-related) changes in RV FAC and RV areas and to establish reference values in healthy preterm and term neonates. A prospective longitudinal study was conducted in 115 preterm infants (23-28 weeks' gestational age at birth, 500-1,500 g). RV FAC was measured at 24 hours of age, 72 hours of age, and 32 and 36 weeks' postmenstrual age (PMA). The maturational patterns of RVEDA, RVESA, and RV FAC were compared with those in 60 healthy full-term infants in a cross-sectional study (≥37 weeks, 3.5 ± 1 kg), who underwent echocardiography at birth (n = 25) and 1 month of age (n = 35). RVEDA and RVESA were traced in the RV-focused apical four-chamber view, and FAC was calculated using the formula 100 × [(RVEDA - RVESA)/RVEDA)]. Premature infants who developed chronic lung disease or had clinically and hemodynamically significant patent ductus arteriosus were excluded (n = 55) from the reference values. Intra- and interobserver reproducibility analysis was performed. RV FAC ranged from 26% at birth to 35% by 36 weeks' PMA in preterm infants (n = 60) and increased almost 2 times faster in the first month of age compared with healthy term infants (n = 60). Similarly, RVEDA and RVESA increased throughout maturation in both term and preterm infants. RV FAC and RV areas were correlated with weight (r = 0.81, P < .001) but were independent of gestational age at birth (r = 0.3, P = .45). RVEDA and RVESA were correlated with PMA in weeks (r = 0.81, P < .001). RV FAC trended lower in preterm infants with bronchopulmonary dysplasia (P = .04) but was not correlated with size of patent ductus arteriosus (P = .56). There was no difference in RV FAC based on gender or need for mechanical ventilation. This study establishes reference values of RV areas (RVEDA and RVESA) and RV FAC in healthy term and preterm infants and tracks their maturational changes during postnatal development. These measures increase from birth to 36 weeks' PMA, and this is reflective of the postnatal cardiac growth as a contributor to the maturation of cardiac function These measures are also linearly associated with increasing weight throughout maturation. This study suggests that two-dimensional RV FAC can be used as a complementary modality to assess global RV systolic function in neonates and facilitates its incorporation into clinical pediatric and neonatal guidelines.


Main routes of transmission of human immunodeficiency virus (HIV) infection in a family setting in Palermo, Italy.
A cross-sectional seroepidemiologic study was carried out on household contacts and sexual partners of human immunodeficiency virus (HIV) antibody-positive intravenous drug abusers in Palermo, Italy, in 1985 to evaluate factors that influenced HIV transmission. A total of 43 index cases, 36 spouses or heterosexual partners, 28 children, and 55 adult household members were enrolled. None of the household members without sexual contact, who had shared items and facilities and had interacted with the index cases, contracted HIV infection. However, six of 36 sexual partners had antibodies to HIV. It was observed that the risk of HIV infection was significantly associated with the frequency of sexual intercourse with the seropositive partner. Four children were also found to be infected: two had acquired immunodeficiency syndrome-related complex, and the other two were clinically and immunologically normal. Furthermore, one other child had evidence of passive transfer of maternal antibodies. The infection was confined to the younger children (ranging in age from eight months to three years). The data support a high rate of vertical transmission from mother to infant, an intermediate rate of transmission to sexual partners, and no transmission attributable to household contact.


Maternal aldehyde elimination during pregnancy preserves the fetal genome.
Maternal metabolism provides essential nutrients to enable embryonic development. However, both mother and embryo produce reactive metabolites that can damage DNA. Here we discover how the embryo is protected from these genotoxins. Pregnant mice lacking Aldh2, a key enzyme that detoxifies reactive aldehydes, cannot support the development of embryos lacking the Fanconi anemia DNA repair pathway gene Fanca. Remarkably, transferring Aldh2(-/-)Fanca(-/-) embryos into wild-type mothers suppresses developmental defects and rescues embryonic lethality. These rescued neonates have severely depleted hematopoietic stem and progenitor cells, indicating that despite intact maternal aldehyde catabolism, fetal Aldh2 is essential for hematopoiesis. Hence, maternal and fetal aldehyde detoxification protects the developing embryo from DNA damage. Failure of this genome preservation mechanism might explain why birth defects and bone marrow failure occur in Fanconi anemia, and may have implications for fetal well-being in the many women in Southeast Asia that are genetically deficient in ALDH2.


Revisiting interaction in knowledge translation.
Although the study of research utilization is not new, there has been increased emphasis on the topic over the recent past. Science push models that are researcher driven and controlled and demand pull models emphasizing users/decision-maker interests have largely been abandoned in favour of more interactive models that emphasize linkages between researchers and decisionmakers. However, despite these and other theoretical and empirical advances in the area of research utilization, there remains a fundamental gap between the generation of research findings and the application of those findings in practice. Using a case approach, the current study looks at the impact of one particular interaction approach to research translation used by a Canadian funding agency. Results suggest there may be certain conditions under which different levels of decisionmaker involvement in research will be more or less effective. Four attributes are illuminated by the current case study: stakeholder diversity, addressability/actionability of results, finality of study design and methodology, and politicization of results. Future research could test whether these or other variables can be used to specify some of the conditions under which different approaches to interaction in knowledge translation are likely to facilitate research utilization. This work suggests that the efficacy of interaction approaches to research translation may be more limited than current theory proposes and underscores the need for more completely specified models of research utilization that can help address the slow pace of change in this area.


Photoactive assemblies of organic compounds and biomolecules: drug-protein supramolecular systems.
The properties of singlet and triplet excited states are strongly medium-dependent. Hence, these species constitute valuable tools as reporters to probe compartmentalised microenvironments, including drug@protein supramolecular systems. In the present review, the attention is focused on the photophysical properties of the probe drugs (rather than those of the protein chromophores) using transport proteins (serum albumins and α1-acid glycoproteins) as hosts. Specifically, fluorescence measurements allow investigation of the structural and dynamic properties of biomolecules or their complexes. Thus, the emission quantum yields and the decay kinetics of the drug singlet excited states provide key information to determine important parameters such as the stoichiometry of the complex, the binding constant, the relative degrees of occupancy of the different compartments, etc. Application of the FRET concept allows determination of donor-acceptor interchromophoric distances. In addition, anisotropy measurements can be related to the orientation of the drug within the binding sites, where the degrees of freedom for conformational relaxation are restricted. Transient absorption spectroscopy is also a potentially powerful tool to investigate the binding of drugs to proteins, where formation of encapsulated triplet excited states is favoured over other possible processes leading to ionic species (i.e. radical ions), and their photophysical properties are markedly sensitive to the microenvironment experienced within the protein binding sites. Even under aerobic conditions, the triplet lifetimes of protein-complexed drugs are remarkably long, which provides a broad dynamic range for identification of distinct triplet populations or for chiral discrimination. Specific applications of the laser flash photolysis technique include the determination of drug distribution among the bulk solution and the protein binding sites, competition of two types of proteins to bind a drug, occurrence of drug-drug interactions within protein binding sites, enzymatic-like activity of the protein or determination of enantiomeric compositions. The use of proteins as supramolecular hosts modifies the photoreactivity of encapsulated substrates by providing protection against oxygen or other external reagents, by imposing conformational restrictions in the binding pockets, or by influencing the stereochemical outcome. In this review, a selected group of examples is presented including decarboxylation, dehalogenation, nucleophilic addition, dimerisation, oxidation, Norrish type II reaction, photo-Fries rearrangement and 6π electrocyclisation.


Greedy learning of multiple objects in images using robust statistics and factorial learning.
We consider data that are images containing views of multiple objects. Our task is to learn about each of the objects present in the images. This task can be approached as a factorial learning problem, where each image must be explained by instantiating a model for each of the objects present with the correct instantiation parameters. A major problem with learning a factorial model is that as the number of objects increases, there is a combinatorial explosion of the number of configurations that need to be considered. We develop a method to extract object models sequentially from the data by making use of a robust statistical method, thus avoiding the combinatorial explosion, and present results showing successful extraction of objects from real images.


The interaction between signal regulatory protein alpha (SIRPα) and CD47: structure, function, and therapeutic target.
CD47 is a broadly expressed membrane protein that interacts with the myeloid inhibitory immunoreceptor SIRPα (also termed CD172a or SHPS-1). SIRPα is the prototypic member of the SIRP paired receptor family of closely related SIRP proteins. Engagement of SIRPα by CD47 provides a downregulatory signal that inhibits host cell phagocytosis, and CD47 therefore functions as a "don't-eat-me" signal. Here, we discuss recent structural analysis of CD47-SIRPα interactions and implications of this for the function and evolution of SIRPα and paired receptors in general. Furthermore, we review the proposed roles of CD47-SIRPα interactions in phagocytosis, (auto)immunity, and host defense, as well as its potential significance as a therapeutic target in cancer and inflammation and for improving graft survival in xenotransplantation.


Elite male faculty in the life sciences employ fewer women.
Women make up over one-half of all doctoral recipients in biology-related fields but are vastly underrepresented at the faculty level in the life sciences. To explore the current causes of women's underrepresentation in biology, we collected publicly accessible data from university directories and faculty websites about the composition of biology laboratories at leading academic institutions in the United States. We found that male faculty members tended to employ fewer female graduate students and postdoctoral researchers (postdocs) than female faculty members did. Furthermore, elite male faculty--those whose research was funded by the Howard Hughes Medical Institute, who had been elected to the National Academy of Sciences, or who had won a major career award--trained significantly fewer women than other male faculty members. In contrast, elite female faculty did not exhibit a gender bias in employment patterns. New assistant professors at the institutions that we surveyed were largely comprised of postdoctoral researchers from these prominent laboratories, and correspondingly, the laboratories that produced assistant professors had an overabundance of male postdocs. Thus, one cause of the leaky pipeline in biomedical research may be the exclusion of women, or their self-selected absence, from certain high-achieving laboratories.


The REM sleep-memory consolidation hypothesis.
It has been hypothesized that REM (rapid eye movement) sleep has an important role in memory consolidation. The evidence for this hypothesis is reviewed and found to be weak and contradictory. Animal studies correlating changes in REM sleep parameters with learning have produced inconsistent results and are confounded by stress effects. Humans with pharmacological and brain lesion-induced suppression of REM sleep do not show memory deficits, and other human sleep-learning studies have not produced consistent results. The time spent in REM sleep is not correlated with learning ability across humans, nor is there a positive relation between REM sleep time or intensity and encephalization across species. Although sleep is clearly important for optimum acquisition and performance of learned tasks, a major role in memory consolidation is unproven.


Magnetocardiographic study of ventricular repolarization in hypertensive patients with and without left ventricular hypertrophy.
Previous studies in magnetically shielded rooms have shown that magnetocardiographic (MCG) mapping can be useful to detect early signs of left ventricular hypertrophy (LVH). The aim of this study was to evaluate ventricular repolarization parameters in patients with essential hypertension, associated or not with LVH, by means of unshielded multichannel MCG mapping. 31 patients with pharmacologically treated essential hypertension (average BP systolic: 147.8+/-11.2, diastolic: 92.2+/-4.9) since 6.5+/-5.6 years, 13 without and 18 with evidence of LVH (4 by ECG, 11 by echocardiography, and 3 at both), were studied with a 36-channel MCG system (sensitivity of 20 fT/square root of Hz1/2) and with 12-lead ECG, in an unshielded hospital setting. To assess ventricular repolarization, HR-corrected, QTend, JTpeak, JTend, Tpeak-end intervals and QT dispersion (QTd) were measured from both MCG and ECG waveforms. The magnetic field gradient orientation (alpha angle) during the ST interval and at the Tpeak was also computed. 20 normal age-matched volunteers were used for comparison. As compared to normal volunteers, MCG JTend, QTend, Tpeak-end and QTd were significantly longer in hypertensive patients. The difference was not significant, if only patients with essential hypertension but no LVH were considered. The magnetic field alpha angle during the ST was significantly abnormal in patients with essential hypertension (p < 0.01). In patients with essential hypertension, MCG detects alterations of ventricular repolarization, not evidenced by 12-lead ECG.


Search for the supersymmetric partner of the top quark in dilepton events from pp collisions at square root of s=1.8 TeV.
We have searched for pair production of the supersymmetric partner of the top quark (stop) in 107 pb(-1) of pp collisions at square root of s=1.8 TeV collected by the Collider Detector at Fermilab (CDF). Each stop is assumed to decay into a lepton, bottom quark, and supersymmetric neutrino. Such a scenario would give rise to events with two leptons, two hadronic jets, and a substantial imbalance of transverse energy. No evidence of such a stop signal has been found. We exclude stop masses in the region (80</=m(t)</=135 GeV/c(2)) in the mass plane of stop versus sneutrino.


Selective recruitment of arrestin-3 to clathrin coated pits upon stimulation of G protein-coupled receptors.
Non-visual arrestins (arrestin-2 and arrestin-3) play critical roles in the desensitization and internalization of many G protein-coupled receptors. In vitro experiments have shown that both non-visual arrestins bind with high and approximately comparable affinities to activated, phosphorylated forms of receptors. They also exhibit high affinity binding, again of comparable magnitude, to clathrin. Further, agonist-promoted internalization of many receptors has been found to be stimulated by exogenous over-expression of either arrestin2 or arrestin3. The existence of multiple arrestins raises the question whether stimulated receptors are selective for a specific endogenous arrestin under more physiological conditions. Here we address this question in RBL-2H3 cells, a cell line that expresses comparable levels of endogenous arrestin-2 and arrestin-3. When (beta)(2)-adrenergic receptors are stably expressed in these cells the receptors internalize efficiently following agonist stimulation. However, by immunofluorescence microscopy we determine that only arrestin-3, but not arrestin-2, is rapidly recruited to clathrin coated pits upon receptor stimulation. Similarly, in RBL-2H3 cells that stably express physiological levels of m1AChR, the addition of carbachol selectively induces the localization of arrestin-3, but not arrestin-2, to coated pits. Thus, this work demonstrates coupling of G protein-coupled receptors to a specific non-visual arrestin in an in vivo setting.


Paranoia as a deficit in non-social belief updating.
Paranoia is the belief that harm is intended by others. It may arise from selective pressures to infer and avoid social threats, particularly in ambiguous or changing circumstances. We propose that uncertainty may be sufficient to elicit learning differences in paranoid individuals, without social threat. We used reversal learning behavior and computational modeling to estimate belief updating across individuals with and without mental illness, online participants, and rats chronically exposed to methamphetamine, an elicitor of paranoia in humans. Paranoia is associated with a stronger prior on volatility, accompanied by elevated sensitivity to perceived changes in the task environment. Methamphetamine exposure in rats recapitulates this impaired uncertainty-driven belief updating and rigid anticipation of a volatile environment. Our work provides evidence of fundamental, domain-general learning differences in paranoid individuals. This paradigm enables further assessment of the interplay between uncertainty and belief-updating across individuals and species.


Molecular dynamics simulations of the helical antimicrobial peptide ovispirin-1 in a zwitterionic dodecylphosphocholine micelle: insights into host-cell toxicity.
We have carried out a 40-ns all-atom molecular dynamics simulation of the helical antimicrobial peptide ovispirin-1 (OVIS) in a zwitterionic diphosphocholine (DPC) micelle. The DPC micelle serves as an economical and effective model for a cellular membrane owing to the presence of a choline headgroup, which resembles those of membrane phospholipids. OVIS, which was initially placed along a micelle diameter, diffuses out to the water-DPC interface, and the simulation stabilizes to an interface-bound steady state in 40 ns. The helical content of the peptide marginally increases in the process. The final conformation, orientation, and the structure of OVIS are in excellent agreement with the experimentally observed properties of the peptide in the presence of lipid bilayers composed of 75% zwitterionic lipids. The amphipathic peptide binds to the micelle with its hydrophobic face buried in the micellar core and the polar side chains protruding into the aqueous phase. There is overwhelming evidence that points to the significant and indispensable participation of hydrophobic residues in binding to the zwitterionic interface. The simulation starts with a conformation that is unbiased toward the final experimentally known binding state of the peptide. The ability of the model to reproduce experimental binding states despite this starting conformation is encouraging.


The design of mineralised hard tissues for their mechanical functions.
Most hard tissues have as their primary purpose to be stiff. Outside the arthropods, mineralisation of a soft organic matrix is the almost universal method of producing high stiffness. However, stiffening brings with it the undesirable mechanical result of brittleness (lack of toughness). The mineralisation of some tissues, such as bone and dentine, can be modified rather easily, in evolutionary terms, to produce the optimum mix of stiffness with bending strength (which, except at the highest mineralisations, go together) on one hand and toughness on the other hand. However, in most other tissues, such as mollusc shell, echinoderm skeleton, brachiopod shell, barnacle shell and enamel, mineralisation is almost all-or-none, and no subtle gradations seem possible. In such cases, other features, such as architecture, must be modified to produce a useful skeleton. Not only the mechanical properties of the skeletal tissue, but its cost, mass and time taken for production will, biologists tend to assume, be balanced by natural selection to produce a satisfactory result. However, such complexity makes it difficult to be sure that we understand the extent to which mineralised skeletal materials are the best possible solution to the problems facing the animals and that we are not just telling 'Just-So' stories. Furthermore, there are some skeletal materials that do not seem to make much sense at the moment, although no doubt all will become clear eventually.


Frameshift signal transplantation and the unambiguous analysis of mutations in the yeast retrotransposon Ty1 Gag-Pol overlap region.
The yeast retrotransposon Ty1 encodes a 7-nucleotide RNA sequence that directs a programmed, +1 ribosomal frameshifting event required for Gag-Pol translation and retrotransposition. We report mutations that block frameshifting, which can be suppressed in cis by "transplanting" the frameshift signal to a position upstream of its native location. These "frameshift transplant" mutants transpose with only a modest decrease in efficiency, suggesting that the location of the frameshift signal in a functional Ty1 element may vary. The genomic architecture of Ty1 is such that Gag, Ty1 PR (PR), and the Gag-derived p4 peptide share a common sequence. The functional independence of the movement of the frameshift signal to a new location within the Ty1 element is used to unambiguously attribute the effect of mutations deleterious to transposition in this region of overlapping coding sequences to effects on the Ty1 (PR). This work defines the amino terminus of the Ty1 PR and introduces a new technique for studying viral genome organization.


Choosing for you: Diminished self-other discrepancies in financial decisions under risk in the elderly.
Many older adults hold powerful positions in governments and corporate boards throughout the world. Accordingly, older adults often have to make important financial decisions on behalf of others under risk. Although it is common to observe younger adults taking more risks when making financial decisions for others, it is unclear if older adults exhibit the same self-other discrepancies. Here, we conducted 2 studies (88 and 124 participants, respectively) to examine self-other discrepancies in financial decision making under risk in older adults. We focused on 3 aspects of financial decision making: loss aversion (a tendency to weight potential losses more strongly than potential gains), risk-aversion asymmetry (a tendency to be risk-averse for potential gains and risk-seeking for potential losses), and risk preferences separately in gain and loss domains. Using computational modeling and behavioral economics tasks, we found weaker self-other discrepancies in older adults (compared with younger adults) across all 3 aspects. We also replicated the age differences in self-other discrepancies in loss aversion across 2 largely nonoverlapping cohorts. Thus, it appears that when making financial decisions on behalf of others, older adults, relative to younger adults, have a stronger disposition to regard others' financial outcomes as important as their own. (PsycINFO Database Record


[Silent pulmonary aspiration of barium after esophagogastroduodenography and subtotal gastrectomy].
We describe a 66-year-old patient who underwent subtotal gastrectomy and omentectomy for gastric adenocarcinoma six days after esophago-gastro-duodenography with barium contrast medium. In the postoperative period a series of chest films showed bronchogram due to aspiration of the contrast medium from the stomach, and finally multiple cottony infiltrates bilaterally. The patient had no respiratory symptoms and 6 days after admission to the intensive care recovery unit, he was moved to the general surgery ward for observation. Follow-up later passed to the outpatient with satisfactory evolution. We conclude that pulmonary aspiration occurred after extubation in the operating theater, since during the operation the patient remained stable hemodynamically and respiratorily. This case corroborates the theory of the existence of silent aspiration without pulmonary complications and the need to adjust clinical treatment, which can be conservative when, once the process is diagnosed, no symptoms present.


Genetic modifiers of menopausal hormone replacement therapy and breast cancer risk: a genome-wide interaction study.
Women using menopausal hormone therapy (MHT) are at increased risk of developing breast cancer (BC). To detect genetic modifiers of the association between current use of MHT and BC risk, we conducted a meta-analysis of four genome-wide case-only studies followed by replication in 11 case-control studies. We used a case-only design to assess interactions between single-nucleotide polymorphisms (SNPs) and current MHT use on risk of overall and lobular BC. The discovery stage included 2920 cases (541 lobular) from four genome-wide association studies. The top 1391 SNPs showing P values for interaction (Pint) <3.0 × 10(-3) were selected for replication using pooled case-control data from 11 studies of the Breast Cancer Association Consortium, including 7689 cases (676 lobular) and 9266 controls. Fixed-effects meta-analysis was used to derive combined Pint. No SNP reached genome-wide significance in either the discovery or combined stage. We observed effect modification of current MHT use on overall BC risk by two SNPs on chr13 near POMP (combined Pint≤8.9 × 10(-6)), two SNPs in SLC25A21 (combined Pint≤4.8 × 10(-5)), and three SNPs in PLCG2 (combined Pint≤4.5 × 10(-5)). The association between lobular BC risk was potentially modified by one SNP in TMEFF2 (combined Pint≤2.7 × 10(-5)), one SNP in CD80 (combined Pint≤8.2 × 10(-6)), three SNPs on chr17 near TMEM132E (combined Pint≤2.2×10(-6)), and two SNPs on chr18 near SLC25A52 (combined Pint≤4.6 × 10(-5)). In conclusion, polymorphisms in genes related to solute transportation in mitochondria, transmembrane signaling, and immune cell activation are potentially modifying BC risk associated with current use of MHT. These findings warrant replication in independent studies.


Oncogenic B-RAF(V600E) signaling induces the T-Box3 transcriptional repressor to repress E-cadherin and enhance melanoma cell invasion.
Approximately 50% of melanomas require oncogenic B-RAF(V600E) signaling for proliferation, survival, and metastasis, and the use of highly selective B-RAF inhibitors has yielded remarkable, although short-term, clinical responses. Reactivation of signaling downstream of B-RAF is frequently associated with acquired resistance to B-RAF inhibitors, and the identification of B-RAF targets may therefore provide new strategies for managing melanoma. In this report, we applied whole-genome expression analyses to reveal that oncogenic B-RAF(V600E) regulates genes associated with epithelial-mesenchymal transition in normal cutaneous human melanocytes. Most prominent was the B-RAF-mediated transcriptional repression of E-cadherin, a keratinocyte-melanoma adhesion molecule whose loss is intimately associated with melanoma invasion and metastasis. Here we identify a link between oncogenic B-RAF, the transcriptional repressor Tbx3, and E-cadherin. We show that B-RAF(V600E) induces the expression of Tbx3, which potently represses E-cadherin expression in melanocytes and melanoma cells. Tbx3 expression is normally restricted to developmental embryonic tissues and promoting cell motility, but it is also aberrantly increased in various cancers and has been linked to tumor cell invasion and metastasis. We propose that this B-RAF/Tbx3/E-cadherin pathway has a critical role in promoting the metastasis of B-RAF-mutant melanomas.


Equivalent induction of telomerase-specific cytotoxic T lymphocytes from tumor-bearing patients and healthy individuals.
Although high frequencies of T lymphocytes specific for certain tumor-associated antigens have been detected in some cancer patients, increasing evidence suggests that these T cells may be functionally defective in vivo and fail to induce meaningful clinical responses. One strategy to overcome this limitation is to target novel antigens that are ignored during the natural antitumor immune response but are nevertheless capable of triggering effector T-cell responses against tumors after optimal presentation by antigen-presenting cells. Here, we show that the telomerase catalytic subunit (hTERT)-a nearly universal tumor antigen identified by epitope deduction rather than from patient immune responses-is immunologically ignored by patients despite progressive tumor burden. Nevertheless, HLA-A2-restricted CTLs against hTERT are equivalently induced ex vivo from patients and healthy individuals and efficiently kill human tumor cell lines and primary tumors. Thus, telomerase-specific T cells from cancer patients are spared functional inactivation because of immunological ignorance. These findings support clinical efforts to target the hTERT as a tumor antigen with broad therapeutic potential.


Alcohol screening and brief interventions for offenders in the probation setting (SIPS Trial): a pragmatic multicentre cluster randomized controlled trial.
To evaluate the effectiveness of different brief intervention strategies at reducing hazardous or harmful drinking in the probation setting. Offender managers were randomized to three interventions, each of which built on the previous one: feedback on screening outcome and a client information leaflet control group, 5 min of structured brief advice and 20 min of brief lifestyle counselling. A pragmatic multicentre factorial cluster randomized controlled trial. The primary outcome was self-reported hazardous or harmful drinking status measured by Alcohol Use Disorders Identification Test (AUDIT) at 6 months (negative status was a score of <8). Secondary outcomes were AUDIT status at 12 months, experience of alcohol-related problems, health utility, service utilization, readiness to change and reduction in conviction rates. Follow-up rates were 68% at 6 months and 60% at 12 months. At both time points, there was no significant advantage of more intensive interventions compared with the control group in terms of AUDIT status. Those in the brief advice and brief lifestyle counselling intervention groups were statistically significantly less likely to reoffend (36 and 38%, respectively) than those in the client information leaflet group (50%) in the year following intervention. Brief advice or brief lifestyle counselling provided no additional benefit in reducing hazardous or harmful drinking compared with feedback on screening outcome and a client information leaflet. The impact of more intensive brief intervention on reoffending warrants further research.


Hunger artists: yeast adapted to carbon limitation show trade-offs under carbon sufficiency.
As organisms adaptively evolve to a new environment, selection results in the improvement of certain traits, bringing about an increase in fitness. Trade-offs may result from this process if function in other traits is reduced in alternative environments either by the adaptive mutations themselves or by the accumulation of neutral mutations elsewhere in the genome. Though the cost of adaptation has long been a fundamental premise in evolutionary biology, the existence of and molecular basis for trade-offs in alternative environments are not well-established. Here, we show that yeast evolved under aerobic glucose limitation show surprisingly few trade-offs when cultured in other carbon-limited environments, under either aerobic or anaerobic conditions. However, while adaptive clones consistently outperform their common ancestor under carbon limiting conditions, in some cases they perform less well than their ancestor in aerobic, carbon-rich environments, indicating that trade-offs can appear when resources are non-limiting. To more deeply understand how adaptation to one condition affects performance in others, we determined steady-state transcript abundance of adaptive clones grown under diverse conditions and performed whole-genome sequencing to identify mutations that distinguish them from one another and from their common ancestor. We identified mutations in genes involved in glucose sensing, signaling, and transport, which, when considered in the context of the expression data, help explain their adaptation to carbon poor environments. However, different sets of mutations in each independently evolved clone indicate that multiple mutational paths lead to the adaptive phenotype. We conclude that yeasts that evolve high fitness under one resource-limiting condition also become more fit under other resource-limiting conditions, but may pay a fitness cost when those same resources are abundant.


Strong and weak male mutation bias at different sites in the primate genomes: insights from the human-chimpanzee comparison.
Male mutation bias is a higher mutation rate in males than in females thought to result from the greater number of germ line cell divisions in males. If errors in DNA replication cause most mutations, then the magnitude of male mutation bias, measured as the male-to-female mutation rate ratio (alpha), should reflect the relative excess of male versus female germ line cell divisions. Evolutionary rates averaged among all sites in a sequence and compared between mammalian sex chromosomes were shown to be indeed higher in males than in females. However, it is presently unknown whether individual classes of substitutions exhibit such bias. To address this issue, we investigated male mutation bias separately at non-CpG and CpG sites using human-chimpanzee whole-genome alignments. We observed strong male mutation bias at non-CpG sites: alpha in the X-autosome comparison was approximately 6-7, which was similar to the male-to-female ratio in the number of germ line cell divisions. In contrast, mutations at CpG sites exhibited weak male mutation bias: alpha in the X-autosome comparison was only approximately 2-3. This is consistent with the methylation-induced and replication-independent mechanism of CpG transitions, which constitute the majority of mutations at CpG sites. Interestingly, our study also indicated weak male mutation bias for transversions at CpG sites, implying a spontaneous mechanism largely not associated with replication. Male mutation bias was equally strong at CpG and non-CpG sites located within unmethylated "CpG islands," suggesting the replication-dependent origin of these mutations. Thus, we found that the strength of male mutation bias is nonuniform in the primate genomes. Importantly, we discovered that male mutation bias depends on the proportion of CpG sites in the loci compared. This might explain the differences in the magnitude of primate male mutation bias observed among studies.


Pyloric stenosis in the Oxford Record Linkage Study area.
The files of the Oxford Record Linkage Study were employed to identify 220 infants presenting with infantile hypertrophic pyloric stenosis (IHPS) in the 6-year period 1966 to 1971. Information on these infants was obtained from birth certificates and maternity notes. The overall incidence was 2.5 per 1000 livebirths. There was a distinct seasonal variation, with highest incidence to infants born in the third quarter of the year as well as variation in incidence with area: the cities had much lower rates of IHPS than the adjacent rural or small urban areas. It was shown that the rates in the south and east of the area studied were far greater than in the north and west. In the present study there was no excess of primiparae, the peak maternal age group was 20 to 24; there was a slight excess of parents of social classes I and II; and a significant association with mothers who were Rhesus negative. The rate of IHPS among sibs was 85 per 1000. Though there was the usual correlation with the male sex (M:F ratio = 5.5:1), there was no variation with birthweight and only among the females was an association found with prolonged gestation. There appeared to be an inverse relation between gestation and age on admission to hospital.


A constitutive model for a maturing fibrin network.
Blood clot formation is crucial to maintain normal physiological conditions but at the same time involved in many diseases. The mechanical properties of the blood clot are important for its functioning but complicated due to the many processes involved. The main structural component of the blood clot is fibrin, a fibrous network that forms within the blood clot, thereby increasing its mechanical rigidity. A constitutive model for the maturing fibrin network is developed that captures the evolving mechanical properties. The model describes the fibrin network as a network of fibers that become thicker in time. Model parameters are related to the structural properties of the network, being the fiber length, bending stiffness, and mass-length ratio. Results are compared with rheometry experiments in which the network maturation is followed in time for various loading frequencies and fibrinogen concentrations. Three parameters are used to capture the mechanical behavior including the mass-length ratio. This parameter agrees with values determined using turbidimetry experiments and is subsequently used to derive the number of protofibrils and fiber radius. The strength of the model is that it describes the mechanical properties of the maturing fibrin network based on it structural quantities. At the same time the model is relatively simple, which makes it suitable for advanced numerical simulations of blood clot formation during flow in blood vessels.


Practice guideline for the management of infants and children 0 to 36 months of age with fever without source. Agency for Health Care Policy and Research.
To develop guidelines for the care of infants and children from birth to 36 months of age with fever without source. An expert panel of senior academic faculty with expertise in pediatrics and infectious diseases or emergency medicine. A comprehensive literature search was used to identify all publications pertinent to the management of the febrile child. When appropriate, meta-analysis was used to combine the results of multiple studies. One or more specific management strategies were proposed for each of the decision nodes in draft management algorithms. The draft algorithms, selected publications, and the meta-analyses were provided to the panel, which determined the final guidelines using the modified Delphi technique. All toxic-appearing infants and children and all febrile infants less than 28 days of age should be hospitalized for parenteral antibiotic therapy. Febrile infants 28 to 90 days of age defined at low risk by specific clinical and laboratory criteria may be managed as outpatients if close follow-up is assured. Older children with fever less than 39.0 C without source need no laboratory tests or antibiotics. Children 3 to 36 months of age with fever of 39.0 C or more and whose WBC count is 15,000/mm3 or more should have a blood culture and be treated with antibiotics pending culture results. Urine cultures should be obtained from all boys 6 months of age or less and all girls 2 years of age or less who are treated with antibiotics. These guidelines do not eliminate all risk or strictly confine antibiotic treatment to children likely to have occult bacteremia. Physicians may individualize therapy based on clinical circumstances or adopt a variation of these guidelines based on a different interpretation of the evidence.


Tuberculosis infection and disease in children living in households of Filipino patients with tuberculosis: a preliminary report.
DOTS Clinic with a DOTS-Plus pilot project for the management of multidrug-resistant tuberculosis (MDR-TB) in a high burden country. To determine the prevalence of tuberculosis (TB) infection and disease among pediatric household contacts of patients with pulmonary TB (PTB). Cross-sectional study. One hundred and fifty-three children aged 0-15 years in the households of 62 bacteriologically confirmed PTB patients, including 44 with MDR-TB, were studied. BCG scars were noted, and tuberculin skin test (TST), screening chest radiography, and sputum or gastric aspirate smear and culture for Mycobacterium tuberculosis in those with radiographic findings suggestive of PTB were done. For children in this study, the prevalences of latent TB infection (LTBI), radiographically diagnosed pulmonary TB, and bacillary pulmonary TB were 69.2%, 3.3%, and 0.65%, respectively. Only age > or = 5 years was found to be a significant predictor of LTBI (OR 3.17, 95%CI 1.43-7.01). Contact investigation for active case-finding and early treatment of TB in children from households of patients with active PTB is essential for TB control. Further study on a more precise definition of TB infection and strategies for control in this population will be pursued.


Mechanisms of action of antithymocyte globulin: old dogs with new tricks!
Understanding the mechanisms underlying immunological tolerance is the key to successful transplantation. Recent findings demonstrate that polyclonal antibodies such as the antithymocyte globulins preparations can provide wide spectrum immunomodulation, suggesting that their use in the immunosuppression therapeutic armamentarium may help in reducing the incidence of organ rejection and improving patients' outcome after hematopoietic stem cell transplantation.


Relationship between cardiac output and oxygen consumption during upright cycle exercise in healthy humans.
The relationship between cardiac output (CardOut) and oxygen consumption (VO2) during exercise has generally been assumed to be linear. To test this assumption, we studied 72 healthy subjects using a graded, 2-min cycle-ergometry exercise test to maximum while measuring gas exchange continuously and CardOut at the end of each stage, the latter using an open-circuit gas technique. Data for VO2 and CardOut at each stage were fit to a quadratic expression y = a + (b.VO2) + (c.VO2(2)), and statistical significance of the quadratic c term was determined in each subject. Subjects were then divided into two groups: those with statistically significant negative quadratic term ("negative curvature group," n = 25) and those with either nonsignificant quadratic term or c significantly > 0 ("non-negative curvature group," n = 47, 2 with c significantly > 0). We found the negative curvature group had significantly higher maximal VO2/kg (median 37.9 vs. 32.4 ml x min(-1) x kg(-1); P = 0.03) higher resting stroke volume (SV; median 77 vs. 60 ml; P = 0.04), lower resting heart rate (HR; median 72 vs. 82 beats/min, P = 0.04), and higher tissue oxygen extraction at maximal exercise (17.1 +/- 2.2 vs 15.5 +/- 2.1 ml/100 ml; P < 0.01), with tendencies for higher maximal CardOut and SV. We also found the HR vs. VO2 relationship to be negatively curved, with negative curvature in HR associated with the negative curvature in CardOut (P < 0.05), suggesting the curvature in the CardOut vs. VO2 relationship was secondary to curvature in HR vs. VO2. We conclude that the CardOut vs. VO2 relationship is not always linear, and negative curvature in the relationship is associated with higher fitness levels in normal, non-elite-athletic subjects.


Calcium-induced asymmetrical beating of triton-demembranated sea urchin sperm flagella.
Asymmetrical bending waves can be obtained by reactivating demembranated sea urchin spermatozoa at high Ca2+ concentrations. Moving-film flash photography shows that asymmetrical flagellar bending waves are associated with premature termination of the growth of the bends in one direction (the reverse bends) while the bends in the opposite direction (the principal bends) grow for one full beat cycle, and with unequal rates of growth of principal and reverse bends. The relative proportions of these two components of asymmetry are highly variable. The increased angle in the principal bend is compensated by a decreased angle in the reverse bend, so that there is no change in mean bend angle; the wavelength and beat frequency are also independent of the degree of asymmetry. This new information is still insufficient to identify a particular mechanism for Ca2+-induced asymmetry. When a developing bend stops growing before initiation of growth of a new bend in the same direction, a modification of the sliding between tubules in the distal portion of the flagellum is required. This modification can be described as a superposition of synchronous sliding on the metachronous sliding associated with propagating bending waves. Synchronous sliding is particularly evident in highly asymmetrical flagella, but is probably not the cause of asymmetry. The control of metachronous sliding appears to be unaffected by the superposition of synchronous sliding.


Radioactive holmium phosphate microspheres for cancer treatment.
The aim of this study was the development of radioactive holmium phosphate microspheres (HoPO4-MS) with a high holmium content and that are stable in human serum for selective internal radiation therapy (SIRT) of liver cancer. To this end, holmium acetylacetonate microspheres (HoAcAc-MS) were prepared (34.2 ± 1.0 µm in diameter, holmium content of 46.2 ± 0.8 and density of 1.7 g/cm3) via an emulsification and solvent evaporation method. The concentration of HoAcAc in the organic solvent, the temperature of emulsification and the stirring speed were varied for the preparation of the HoAcAc-MS to obtain microspheres with different diameters ranging from 11 to 35 µm. Subsequently, the AcAc ligands of the HoAcAc-MS were replaced by phosphate ions by simply incubating neutron irradiated HoAcAc-MS in a phosphate buffer solution (0.116 M, pH 4.2) to yield radioactive HoPO4-MS. The obtained microspheres were analyzed using different techniques such as SEM-EDS, ICP-OES and HPLC. The prepared HoPO4-MS (29.5 ± 1.2 µm in diameter and a density of 3.1 g/cm3) present an even higher holmium content (52 wt%) than the HoAcAc-MS precursor (46 wt%). Finally, the stability of the HoPO4-MS was tested by incubation in human serum at 37 °C which showed no visible changes of the microspheres morphology and only 0.1% of holmium release was observed during the 2 weeks period of incubation. In conclusion, this study shows that stable radioactive HoPO4-MS can be prepared with suitable properties to be used for cancer therapy.


Structural and functional characterization of second-coordination sphere mutants of soybean lipoxygenase-1.
Lipoxygenases are an important class of non-heme iron enzymes that catalyze the hydroperoxidation of unsaturated fatty acids. The details of the enzymatic mechanism of lipoxygenases are still not well understood. This study utilizes a combination of kinetic and structural probes to relate the lipoxygenase mechanism of action with structural modifications of the iron's second coordination sphere. The second coordination sphere consists of Gln(495) and Gln(697), which form a hydrogen bond network between the substrate cavity and the first coordination sphere (Asn(694)). In this investigation, we compared the kinetic and structural properties of four mutants (Q495E, Q495A, Q697N, and Q697E) with those of wild-type soybean lipoxygenase-1 and determined that changes in the second coordination sphere affected the enzymatic activity by hydrogen bond rearrangement and substrate positioning through interaction with Gln(495). The nature of the C-H bond cleavage event remained unchanged, which demonstrates that the mutations have not affected the mechanism of hydrogen atom tunneling. The unusual and dramatic inverse solvent isotope effect (SIE) observed for the Q697E mutant indicated that an Fe(III)-OH(-) is the active site base. A new transition state model for hydrogen atom abstraction is proposed.


No Effect of Commercial Cognitive Training on Brain Activity, Choice Behavior, or Cognitive Performance.
Increased preference for immediate over delayed rewards and for risky over certain rewards has been associated with unhealthy behavioral choices. Motivated by evidence that enhanced cognitive control can shift choice behavior away from immediate and risky rewards, we tested whether training executive cognitive function could influence choice behavior and brain responses. In this randomized controlled trial, 128 young adults (71 male, 57 female) participated in 10 weeks of training with either a commercial web-based cognitive training program or web-based video games that do not specifically target executive function or adapt the level of difficulty throughout training. Pretraining and post-training, participants completed cognitive assessments and functional magnetic resonance imaging during performance of the following validated decision-making tasks: delay discounting (choices between smaller rewards now vs larger rewards in the future) and risk sensitivity (choices between larger riskier rewards vs smaller certain rewards). Contrary to our hypothesis, we found no evidence that cognitive training influences neural activity during decision-making; nor did we find effects of cognitive training on measures of delay discounting or risk sensitivity. Participants in the commercial training condition improved with practice on the specific tasks they performed during training, but participants in both conditions showed similar improvement on standardized cognitive measures over time. Moreover, the degree of improvement was comparable to that observed in individuals who were reassessed without any training whatsoever. Commercial adaptive cognitive training appears to have no benefits in healthy young adults above those of standard video games for measures of brain activity, choice behavior, or cognitive performance.SIGNIFICANCE STATEMENT Engagement of neural regions and circuits important in executive cognitive function can bias behavioral choices away from immediate rewards. Activity in these regions may be enhanced through adaptive cognitive training. Commercial brain training programs claim to improve a broad range of mental processes; however, evidence for transfer beyond trained tasks is mixed. We undertook the first randomized controlled trial of the effects of commercial adaptive cognitive training (Lumosity) on neural activity and decision-making in young adults (N = 128) compared with an active control (playing on-line video games). We found no evidence for relative benefits of cognitive training with respect to changes in decision-making behavior or brain response, or for cognitive task performance beyond those specifically trained.


Safety and efficacy of the subclavian approach for transcatheter aortic valve implantation with the CoreValve revalving system.
Transcatheter aortic valve implantation (TAVI) is a new option for patients with severe aortic stenosis at high surgical risk. The standard retrograde approach through the femoral artery is contraindicated in case of unfavorable iliofemoral anatomy or extensive disease. In these patients, a trans-subclavian approach may be feasible. Between June 2007 and July 2009, TAVI with the CoreValve bioprosthesis (Medtronic, Minneapolis, Minn) was performed in 514 consecutive patients at 13 Italian hospitals, using the subclavian approach in 54 cases. The median logistic EuroSCORE was significantly higher in the subclavian (19.4; interquartile range, 12.5 to 29.8) versus femoral group (25.3; interquartile range, 15.1 to 36.6) (P=0.03), as well as the rate of comorbidities. Procedural success was obtained in 100% versus 98.4% of the subclavian versus femoral groups, respectively (P=0.62), with intraprocedural mortality of 0% versus 0.9% (P=1.00). The most common in-hospital complications were a new left bundle-branch block (22.4%) and the need for pacemaker (16.3%). No specific complications for the subclavian access (vessel rupture, vertebral or internal mammary ischemia) were reported. The learning curve for the subclavian approach led to a wider use of local anesthesia. Thirty-day mortality was 0% versus 6.1% in the subclavian versus femoral groups, respectively (P=0.13). Six-month mortality rate was 9.4% versus 15.8% (P=0.44), whereas valve-related adverse events were 13.6% versus 13.9% (P=0.79). TAVI through the subclavian approach appeared feasible and safe, with excellent procedural success and low in-hospital complication rates. This new technique allows patients with contraindications to the femoral approach to be treated with TAVI.


High-Content Analysis of CRISPR-Cas9 Gene-Edited Human Embryonic Stem Cells.
CRISPR-Cas9 gene editing of human cells and tissues holds much promise to advance medicine and biology, but standard editing methods require weeks to months of reagent preparation and selection where much or all of the initial edited samples are destroyed during analysis. ArrayEdit, a simple approach utilizing surface-modified multiwell plates containing one-pot transcribed single-guide RNAs, separates thousands of edited cell populations for automated, live, high-content imaging and analysis. The approach lowers the time and cost of gene editing and produces edited human embryonic stem cells at high efficiencies. Edited genes can be expressed in both pluripotent stem cells and differentiated cells. This preclinical platform adds important capabilities to observe editing and selection in situ within complex structures generated by human cells, ultimately enabling optical and other molecular perturbations in the editing workflow that could refine the specificity and versatility of gene editing.


Validation of a Korean version of the insomnia severity index.
The purposes of this study were to standardize and validate a Korean version of the Insomnia Severity Index (ISI-K), and to evaluate its clinical usefulness. We translated the ISI into Korean and then translated it back into English to check its accuracy. The 614 patients with sleep disorders who were enrolled in this study comprised 169 with primary insomnia, 133 with comorbid insomnia, and 312 with obstructive sleep apnea. All subjects underwent one night of polysomnography (PSG) and completed the Korean versions of both the Pittsburgh Sleep Quality Index (PSQI-K) and the Epworth Sleepiness Scale, as well as the ISI-K. The ISI-K was compared to these sleep scales and various PSG sleep parameters. The internal consistency the ISI-K total score was confirmed by a Cronbach's alpha of 0.92, and the item-to-total-score correlations (item-total correlations) ranged from 0.65 to 0.84, suggesting adequate reliability. The correlation between the ISI-K total score and PSQI-K was 0.84, which suggested adequate convergent validity. Low-to-moderate correlations were obtained between the ISI-K total score and PSG-defined sleep parameters: 0.22 for sleep onset latency, 0.38 for wake after sleep onset, and 0.46 for sleep efficiency. A cutoff score of 15.5 on the ISI-K was optimal for discriminating patients with insomnia. The test-retest scores over a 4-week interval with 34 subjects yielded a correlation coefficient of 0.86, suggesting excellent temporal stability. The findings of this study show that the ISI-K is a reliable and valid instrument for assessing the severity of insomnia in a Korean population.


Human optic nerve fiber count and optic disc size.
In the optic nerve head, the optic nerve fibers are represented by the neuroretinal rim. The rim area showing a high interindividual variability is positively correlated with the optic disc size. This study was performed to address the question of whether, in addition to having a larger neuroretinal rim, eyes with large optic discs also have a higher count of optic nerve fibers compared to eyes with small optic nerve heads. Histologic semithin sections of 72 optic nerves of 56 cornea donors were histomorphometrically evaluated using a computerized image analyzer. The optic nerve fiber count increased significantly (P = 0.01) with enlarging optic disc size. The nerve fiber count was positively correlated with the retrobulbar optic nerve cross section area. It decreased with advancing age, with a mean annual loss of about 4,000 fibers. The nerve fiber density per disc area decreased with increasing optic disc area. Mean and median of the minimal nerve fiber diameter was larger in older subjects. The results may indicate that the optic nerve fiber count, and the anatomic reserve capacity in progressive optic neuropathies, are higher in eyes with large optic discs than in eyes with small optic nerve heads. The optic nerve fiber population decreased with advancing age. This is important for progression, pseudoprogression, and prognosis of optic neuropathies. Optic nerve fiber crowding is more marked in eyes with small optic discs than in eyes with large optic nerve heads. The age-related loss of predominantly small optic nerve fibers can potentiate the optic nerve atrophy in glaucoma and Alzheimer's disease, with both damaging preferentially large axons.


Glycyrrhizin Suppresses RANKL-Induced Osteoclastogenesis and Oxidative Stress Through Inhibiting NF-κB and MAPK and Activating AMPK/Nrf2.
The treatment for osteoporosis involves inhibiting bone resorption and osteoclastogenesis. Glycyrrhizin (GLY) is a triterpenoid saponin glycoside known to be as the most medically efficacious component of the licorice plant. It has strong anti-inflammatory, antioxidant, and antitumor properties. We investigated the effect of GLY on osteoclastogenesis, bone resorption, and intracellular oxidative stress and its molecular mechanisms. In vitro osteoclastogenesis assays were performed using bone marrow monocytes with and without glycyrrhizin. We also evaluated the effects of glycyrrhizin on the secretion of TNF-α, IL-1β, and IL-6 in LPS-stimulated RAW 264.7 cells using ELISA. The effects of glycyrrhizin on the expression of osteoclast-related genes, such as Nfatc1, c-fos, Trap, and cathepsin K (CK), were investigated by RT-PCR. Intracellular reactive oxygen species (ROS) were detected in receptor activator of nuclear factor kappa-Β ligand (RANKL)-stimulated osteoclasts in the presence and absence of glycyrrhizin. During the inhibition of osteoclastogenesis by glycyrrhizin, phosphorylation of AMPK, Nrf2, NF-κB, and MAPK was analyzed using western blotting. Our results showed that glycyrrhizin significantly inhibited RANKL-induced osteoclastogenesis, downregulated the expression of NFATc1, c-fos, TRAP, CK, DC-STAMP, and OSCAR, and inhibited p65, p38, and JNK. Glycyrrhizin was found to significantly decrease the secretion of inflammatory cytokines (TNF-α, IL-1β, and IL-6). Additionally, glycyrrhizin reduced the formation of ROS in osteoclasts by inducing AMPK phosphorylation and nuclear transfer of NRF2, resulting in an upregulation of antioxidant enzymes, such as HO-1, NQO-1, and GCLC. In summary, we found that glycyrrhizin inhibited RANKL-induced osteoclastogenesis. It was also indicated that glycyrrhizin could reduce oxidative stress by inhibiting the MAPK and NF-κB pathways and activating the AMPK/NRF2 signaling. Therefore, glycyrrhizin may be used as an effective therapeutic agent against osteoporosis and bone resorption.


Quantitative evaluation of scattering in optical coherence tomography skin images using the extended Huygens-Fresnel theorem.
An optical properties extraction algorithm is developed based on enhanced Huygens-Fresnel light propagation theorem, to extract the scattering coefficient of a specific region in an optical coherence tomography (OCT) image. The aim is to quantitatively analyze the OCT images. The algorithm is evaluated using a set of phantoms with different concentrations of scatterers, designed based on Mie theory. The algorithm is then used to analyze basal cell carcinoma and healthy eyelid tissues, demonstrating distinguishable differences in the scattering coefficient between these tissues. In this study, we have taken advantage of the simplification introduced by the utilization of a dynamic focus OCT system. This eliminates the need to deconvolve the reflectivity profile with the confocal gate profile, as the sensitivity of the OCT system is constant throughout the axial range.


A randomized, double-blind study of the safety, transmissibility and phenotypic and genotypic stability of cold-adapted influenza virus vaccine.
Live attenuated influenza vaccine (LAIV; FluMist) is a trivalent vaccine containing cold-adapted influenza vaccine viruses that infect and replicate in cells lining the nasopharynx to induce immunity. Recovery of viruses (shedding) is measured by culture of nasal specimens. Shedding of vaccine viruses is not equated with transmission because transmission requires more virus than is detected in many nasal swabs. Previous studies with LAIV did not detect transmission to close contacts. The primary objective of this study was to estimate the probability of transmission to placebo contacts in a day care setting. One hundred ninety-seven healthy children aged 9 to 36 months attending day care were randomized to receive vaccine or placebo. Postvaccination viral shedding, safety, genotype and phenotype of shed viruses and probability of transmission were assessed. Eighty percent of 98 vaccine recipients shed at least one vaccine strain. No clinically significant differences in solicited adverse events attributable to vaccine occurred; safety profiles were similar in both groups. Vaccine virus isolates retained their phenotypic characteristics (cold adaptation and temperature sensitivity) and did not revert at nucleotides known to confer an attenuating phenotype. There was one confirmed transmission of a vaccine strain to a single placebo recipient. According to the Reed-Frost model, the calculated probability of transmission to a child after contact with a single vaccinated child was 0.58% (95% confidence interval, 0-1.7%). There was no increased reactogenicity or other safety concerns in the recipient child. Young children in a day care setting had a high rate of shedding and a low rate of transmission. No clinically significant illness occurred among children who received vaccine or placebo or in the child to whom the vaccine virus was transmitted.


Smoking habits in bipolar and schizophrenic outpatients in southern Israel.
Although rates of cigarette smoking have been found to be higher in schizophrenic and depressed patients than in the general population, data regarding rates in bipolar patients are limited. This study further examines the relationship between bipolar disorder and smoking and compares the rate of smoking in bipolar disorder patients with rates in schizophrenic patients and in the general population. Seventy bipolar patients and 64 schizophrenic patients (diagnosed using DSM-IV criteria) treated at the largest specialized public bipolar and schizophrenia clinics in southern Israel were interviewed regarding their smoking habits. The interview included a questionnaire relating to personal information, past and present smoking, and drug abuse and the Fagerstrom scale for nicotine dependence. Data from these patients were also compared with data from the general Israeli population. Data indicate that the rate of smoking does not appear to differ between bipolar (43.0%) and schizophrenic (45.0%) patients, whereas the rate for both patient groups is higher than that for the general Israeli population (27.5%). Smoking intensity was not found to be different between the 2 groups of patients. Smoking in patients with schizophrenia was suggested to be related to nicotine cholinergic dysfunction, but this suggestion cannot explain the equally high rates of smoking in bipolar patients. Schizophrenia, bipolar disorder, and smoking may all be related to dopamine transmission, and, therefore, dopaminergic interactions may provide a better explanation for the results.


Pre-transfusion management of children with severe malarial anaemia: a randomised controlled trial of intravascular volume expansion.
Symptomatic severe malarial anaemia (SMA) has a high fatality rate of 30-40%; most deaths occur in children awaiting blood transfusion. Blood transfusion services in most of Africa are not capable of delivering adequate supplies of safe blood in a timely manner to critically ill children with SMA. Contrary to widely held belief, hypovolaemia, rather than heart failure, has emerged as a common complication in such children. We examined the safety of pre-transfusion management (PTM) by volume expansion, aimed at stabilizing children and obviating the urgency for blood transfusion. Kenyan children with severe falciparum anaemia (haemoglobin <5 g/dl) and respiratory distress were randomly assigned to 20 ml/kg of 4.5% albumin or 0.9% saline or maintenance only (control) while awaiting blood transfusion. PTM was apparently safe since it did not lead to the development of pulmonary oedema or other adverse events. There was no significant difference in the primary outcome [mean percentage reduction in base excess between admission and 8 h (95% confidence interval)] between the control group 42% (19-66%) albumin group 44% (32-57%) and saline group 36% (16-57%); adjusted analysis of variance F=0.31, P=0.7. However, the number of children requiring emergency interventions was significantly greater in the control group, four of 18 (22%) than the saline group 0 of 20 (P=0.03). We have established the safety of this PTM in children with SMA whilst awaiting blood transfusion at a hospital with an adequate blood-banking program. The impact on mortality should be assessed where blood transfusion services are unable to supply emergency transfusions.


Gene pathways and cell cycle-related genes in cultured avian primordial germ cells.
Primordial germ cells (PGC) from early embryos are applicable to various kinds of research, including the production of transgenic animals. Primordial germ cells eventually migrate and differentiate into germ cells in the gonads, where they settle and rapidly proliferate. However, the proliferation rate of PGC is low in early embryos, and there are many significant pathways that mediate PGC activity. Therefore, in vitro culture of PGC from early embryos with efficient growth factors has been necessary. Recently, we cultured chicken PGC from embryonic d 2.5 with basic fibroblast growth factor and characterized the PGC through analysis of cell morphology, survival, proliferation, and apoptosis. However, large-scale analyses of genes expressed in cultured PGC and the genes involved in associated pathways are limited. The objective of the present investigation was to identify the signaling and metabolic pathways of expressed genes by microarray comparison between PGC and their somatic counterpart, chicken embryonic fibroblasts (CEF). We identified 795 genes that were expressed more predominantly in PGC and 824 genes that were expressed more predominantly in CEF. Among the predominant genes in PGC, 201 were differentially identified in 106 pathways. Among the predominant genes in CEF, 242 were differentially identified in 99 pathways. To further validate the genes involved in at least one candidate pathway, those involved in the cell cycle (12 predominant genes in PGC and 8 predominant genes in CEF) were examined by real-time PCR. To the best of our knowledge, this study is the first to investigate signaling and metabolic pathways in cultured PGC.


Flavopiridol, a novel cyclin-dependent kinase inhibitor, in metastatic renal cancer: a University of Chicago Phase II Consortium study.
Flavopiridol is the first cyclin-dependent kinase (cdk) inhibitor to enter clinical trials. Serum levels of flavopiridol obtained during phase I studies were sufficient to inhibit in vitro cancer cell growth. Because responses were observed in kidney cancer patients in the phase I trials, we performed a phase II trial of flavopiridol in this patient population. Thirty-five minimally pretreated patients were accrued using a standard two-step mechanism. Flavopiridol (50 mg/m(2)/d) was administered by continuous infusion for 72 hours every 2 weeks, and response was evaluated every 8 weeks. Peripheral blood mononuclear cells (PBMCs) were collected at baseline, at completion of drug infusion, and on day 7 of the first therapy cycle, and cell cycle parameters after phytohemagglutinin and interleukin-2 stimulation were assessed. There were two objective responses (response rate = 6%, 95% confidence interval, 1% to 20%). The most common toxicities were asthenia, occurring in 83% of patients (grade 3 or 4 in 9%), and diarrhea, occurring in 77% of patients (grade 3 or 4 in 20%). Also, nine patients (26%) experienced grade 3 or 4 vascular thrombotic events, including one myocardial infarction, two transient neurologic ischemic attacks, four deep venous thrombosis, and two pulmonary emboli. Cell cycle studies did not reveal any effect of flavopiridol on stimulated PBMCs. Flavopiridol, at the dose and schedule administered in this trial, is ineffective in metastatic renal cancer. In addition to the diarrhea observed in phase I studies, we also observed a higher incidence of asthenia and serious vascular thrombotic events than expected.


Comparative in vitro activities of ciprofloxacin, clinafloxacin, gatifloxacin, levofloxacin, moxifloxacin, and trovafloxacin against Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, and Enterobacter aerogenes clinical isolates with alterations in GyrA and ParC proteins.
The in vitro activities of ciprofloxacin, clinafloxacin, gatifloxacin, levofloxacin, moxifloxacin, and trovafloxacin were tested against 72 ciprofloxacin-resistant and 28 ciprofloxacin-susceptible isolates of Klebsiella pneumoniae, Klebsiella oxytoca, Enterobacter cloacae, and Enterobacter aerogenes. Irrespective of the alterations in GyrA and ParC proteins, clinafloxacin exhibited greater activity than all other fluoroquinolones tested against K. pneumoniae and E. aerogenes.


Human RAD 17 Polymorphism at Codon 546 Is Associated with the Risk of Colorectal Cancer.
Human RAD17 acts as an activator of checkpoint signals in response to DNA damage. Here we evaluated the association of hRAD17 Leu546Arg (rs1045051), a missense single nucleotide polymorphism, with the risk of colorectal cancer (CRC) in relation to smoking and alcohol consumption habits in 212 CRC patients and 1,142 cancer-free controls in a case-control study conducted in Japan. The results showed that the hRAD17 Leu/Arg genotype compared to the Leu/Leu genotypes was significantly associated with the protective effect on CRC risk with the adjusted odds ratio (OR) of 0.68 [95% confidence interval (CI): 0.49-0.95, p=0.024], and the males with the Arg/Arg genotype had a greater risk of CRC compared to those with the Leu/Leu and Leu/Arg genotypes (OR=1.87, 95%CI 1.03-3.40, p=0.04). In stratified studies, the protective effect of the Leu/Arg genotype on CRC risk was markedly higher in the light smokers (< 20 pack years) (OR=0.61, 95%CI 0.40-0.94, p=0.024) and the rectal cancer patients (OR=0.49, 95%CI 0.31-0.78, p=0.003). The risk of the Arg/Arg genotype was associated with heavy smoking (≥ 20 pack-years) (OR=2.24, 95%CI 1.09-4.61, p=0.03). These findings suggest that the genetic variant of hRAD17 Leu546Arg polymorphism has a significant effect on CRC susceptibility in Japanese.


Imaging the survival and utility of pre-differentiated allogeneic MSC in ischemic heart.
The aim of the study is to track the survival and utility of mesenchymal stem cells (MSCs) and pre-differentiated MSCs in allogeneic infarcted myocardium. MSCs labeled with green fluorescent protein and luciferase (GFP-Fluc) were characterized by flow cytometry and multi-differentiation. 5-Azacytidine (5-AZ) was employed to induced cardiac differentiation from MSCs. Cardiac markers and immune antigen expression were assessed. Then, pre-differentiated MSCs induced by 5-AZ were intramyocardially injected into allogeneic C57 mice of myocardial infarction, undifferentiated MSCs were transplanted as control. The survival of transplanted cells, immune response and cardiac function of recipients were assessed with bioluminescence imaging, immunohistochemistry and echocardiography, respectively. In vitro results showed that 5-AZ treatment induced cardiac differentiation from MSCs, which also increased their expression of MHC-Ia and MHC-II. After intramyocardial transplantation in allogeneic mice, 5-AZ treated MSCs would rapidly be recognized and excluded by recipients. Meanwhile, a severe infiltration of immune cells could be detected. Though beneficial effects on cardiac function by 5-AZ treated MSCs could be detected, it was short and disappeared within 1 month. In contrast, undifferentiated MSCs were immune-privileged and could survive in allogeneic myocardium for more than 1 month, resulting in a significant improvement on cardiac function.


What Is so Positive about Positive Animal Welfare?-A Critical Review of the Literature.
It is claimed that positive animal welfare (PAW) developed over the last decade in reaction to animal welfare focusing too much on avoiding negatives. However, it remains unclear what PAW adds to the animal welfare literature and to what extent its ideas are new. Through a critical review of the PAW literature, we aim to separate different aspects of PAW and situate it in relation to the traditional animal welfare literature. We find that the core PAW literature is small (n = 10 papers) but links to wider areas of current research interest. The PAW literature is defined by four features: (1) positive emotions which is arguably the most widely acknowledged; (2) positive affective engagement which serves to functionally link positive emotions to goal-directed behavior; (3) quality of life which serves to situate PAW within the context of finding the right balance of positives over negatives; (4) happiness which brings a full life perspective to PAW. While the two first points are already part of welfare research going back decades, the two latter points could be linked to more recent research agendas concerning aggregation and how specific events may affect the ability of animals to make the best of their lives.


Simplification of the pulmonary embolism severity index for prognostication in patients with acute symptomatic pulmonary embolism.
The Pulmonary Embolism Severity Index (PESI) estimates the risk of 30-day mortality in patients with acute pulmonary embolism (PE). We constructed a simplified version of the PESI. The study retrospectively developed a simplified PESI clinical prediction rule for estimating the risk of 30-day mortality in a derivation cohort of Spanish outpatients. Simplified and original PESI performances were compared in the derivation cohort. The simplified PESI underwent retrospective external validation in an independent multinational cohort (Registro Informatizado de la Enfermedad Tromboembólica [RIETE] cohort) of outpatients. In the derivation data set, univariate logistic regression of the original 11 PESI variables led to the removal of variables that did not reach statistical significance and subsequently produced the simplified PESI that contained the variables of age, cancer, chronic cardiopulmonary disease, heart rate, systolic blood pressure, and oxyhemoglobin saturation levels. The prognostic accuracy of the original and simplified PESI scores did not differ (area under the curve, 0.75 [95% confidence interval (CI), 0.69-0.80]). The 305 of 995 patients (30.7%) who were classified as low risk by the simplified PESI had a 30-day mortality of 1.0% (95% CI, 0.0%-2.1%) compared with 10.9% (8.5%-13.2%) in the high-risk group. In the RIETE validation cohort, 2569 of 7106 patients (36.2%) who were classified as low risk by the simplified PESI had a 30-day mortality of 1.1% (95% CI, 0.7%-1.5%) compared with 8.9% (8.1%-9.8%) in the high-risk group. The simplified PESI has similar prognostic accuracy and clinical utility and greater ease of use compared with the original PESI.


In vitro model to study chondrogenic differentiation in tendinopathy.
Treatment of midportion Achilles tendinopathy is hampered by limited knowledge of the pathophysiology. Chondrogenic differentiation of tendon cells might take place in midportion Achilles tendinopathy and could be used as a target for drug treatment. An in vitro model for chondrogenic differentiation would be useful to evaluate existing and future treatment opportunities. A controlled laboratory study. Perioperatively harvested tissue from human midportion Achilles tendinotic lesions and healthy Achilles tendons was analyzed by microscopy and real-time reverse transcription polymerase chain reaction. In vitro chondrogenic differentiation of tendon explants was induced using transforming-growth-factor beta. This model was modulated by removing the chondrogenic stimulus or adding triamcinolone or platelet-rich plasma. Midportion Achilles tendinotic lesions had increased glycosaminoglycan staining and more rounded cell nuclei. Chondrogenic markers (sex-determining region Y)-box9, aggrecan, collagen 2, and RUNT-related transcription factor 2 were upregulated, but collagen 10 was not. Nondegenerative tendon explants cultured on chondrogenic medium had higher expression of aggrecan, collagen 2, and collagen 10 but not (sex-determining region Y)-box9 and RUNT-related transcription factor 2. Removing the chondrogenic stimulus decreased expression of aggrecan, collagen 2, and collagen 10. Both triamcinolone and platelet-rich plasma influenced the chondrogenic gene expression pattern in the in vitro model. Chondrogenic differentiation is present in midportion Achilles tendinopathy. An in vitro model to study this chondrogenic differentiation was developed. This model can be used to investigate chondrogenic differentiation as a possible target for drug treatment, contributing to the development of more successful mechanism-based treatment opportunities.


New azasterols against Trypanosoma brucei: role of 24-sterol methyltransferase in inhibitor action.
A series of azasterol derivatives, designed as potential inhibitors of the Delta(24)-sterol methyltransferase enzyme (24-SMT), were synthesized and evaluated for their activities against parasitic protozoa. Values in the nanomolar range were obtained for 50% effective dose against the Trypanosoma brucei subsp. rhodesiense bloodstream form cultured in vitro. In order to investigate the mode of action, Trypanosoma brucei subsp. brucei 24-SMT was cloned and overexpressed and compounds were assayed for inhibitory activity. None of the inhibitors tested appeared to be active against the enzyme. Sterol composition analysis showed that only cholestane type sterols are present in membranes of bloodstream forms while ergosterol is a major component of procyclic sterol extracts. Interestingly, Northern blot analysis showed the presence of 24-SMT mRNA in both the procyclic and the bloodstream forms of the parasite, although levels of mRNA were threefold lower in the latter. Likewise, Western blot analysis and activity determinations evidenced the existence of active enzyme in both forms of the parasite. We conclude that the designed compounds act at sites other than 24-SMT in Trypanosoma brucei.