Expert consensus for performing right heart catheterisation for suspected pulmonary arterial hypertension in systemic sclerosis: a Delphi consensus study with cluster analysis.
To establish an expert consensus on which criteria are the most appropriate in clinical practice to refer patients with systemic sclerosis (SSc) for right heart catheterisation (RHC) when pulmonary hypertension (PH) is suspected. A three stage internet based Delphi consensus exercise involving worldwide PH experts was designed. In the first stage, a comprehensive list of domains and items combining evidence based indications and expert opinions were obtained. In the second and third stages, experts were asked to rate each item selected in the list. After each of stages 2 and 3, the number of items and criteria were reduced according to a cluster analysis. A literature search and the opinions of 47 experts participating in Delphi stage 1 provided a list of seven domains containing 142 criteria. After stages 2 and 3, these domains and tools were reduced to three domains containing eight tools: clinical (progressive dyspnoea over the past 3 months, unexplained dyspnoea, worsening of WHO dyspnoea functional class, any finding on physical examination suggestive of elevated right heart pressures and any sign of right heart failure), echocardiography (systolic pulmonary artery pressure >45 mm Hg and right ventricle dilation) and pulmonary function tests (diffusion lung capacity for carbon monoxide <50% without pulmonary fibrosis). Among experts in pulmonary arterial hypertension-SSc, a core set of criteria for clinical practice to refer SSc patients for RHC has been defined by Delphi consensus methods. Although these indications are recommended by this expert group to be used as an interim tool, it will be necessary to formally validate the present tools in further studies.


Prevalence and determinants of obesity and dietary habits among adults in rural area, Chile.
This study was undertaken to examine the prevalence of obesity and its determinants among adults in a rural area of Chile. A community-based cross-sectional study was conducted in April-June 2004 in San Carlos (8th region). Height, weight, and waist and hip circumferences were measured for 603 adults (female 447, male 156) aged 20-64 years, and data on socio-economic factors, dietary intake, and dietary habits were obtained by questionnaire interviews. The prevalence of obese individuals was 45.2% among females and 30.1% among males, whereas that of overweight individuals was higher among males than females. Obesity was associated with socio-economic factors for females but not for males. With regard to diet, different patterns between females and males were observed in terms of frequency of food intake, as well as in dietary habits. Our findings of a high prevalence of obese/overweight individuals, together with the characteristics of their diets including changes in the cooking process, suggest that nutrition transition is underway in rural regions as well. Appropriate interventions, therefore, should be introduced to control obesity among women and to enhance health awareness among men throughout the country.


Steroidogenic refractoriness of bovine adrenocortical cells to dibutyryl cyclic AMP.
The long-term effects of dibutyryl cyclic AMP [(Bu)2-cAMP] on steroidogenesis in bovine adrenocortical cells maintained in primary culture were investigated. During the first 36 h, total steroid production by cells incubated with (Bu)2-cAMP increased progressively. Thereafter, however, there was a marked fall in steroid output. During the first 36 h adrenocortical cells incubated in the presence of (Bu)2-cAMP produced substantially more C19-steroids and 17 alpha-hydroxylated C21-steroids than did cells incubated in the absence of (Bu)2-cAMP. By 48 h, however, such steroid secretion by cells incubated in the continued presence of (Bu)2-cAMP declined toward control levels. By contrast, the secretion of corticosterone and 11-deoxycorticosterone was consistently less by cells maintained in the presence of (Bu)2-cAMP than by cells maintained in its absence. These results suggest that refractoriness results, at least in part, from events which occur distal to the formation of cAMP. The action of ACTH and (Bu)2cAMP to promote the secretion of 17 alpha-hydroxylated C21-steroids and C19-steroids, on the other hand, appears to reflect an increase in the rate of cholesterol side-chain cleavage, as well as an increase in 17 alpha-hydroxylase and possibly also 17, 20-lyase activities.


Incendiary Leptin.
Leptin is a hormone released by adipose tissue that plays a key role in the control of energy homeostasis through its binding to leptin receptors (LepR), mainly expressed in the hypothalamus. Most scientific evidence points to leptin's satiating effect being due to its dual capacity to promote the expression of anorexigenic neuropeptides and to reduce orexigenic expression in the hypothalamus. However, it has also been demonstrated that leptin can stimulate (i) thermogenesis in brown adipose tissue (BAT) and (ii) the browning of white adipose tissue (WAT). Since the demonstration of the importance of BAT in humans 10 years ago, its study has aroused great interest, mainly in the improvement of obesity-associated metabolic disorders through the induction of thermogenesis. Consequently, several strategies targeting BAT activation (mainly in rodent models) have demonstrated great potential to improve hyperlipidemias, hepatic steatosis, insulin resistance and weight gain, leading to an overall healthier metabolic profile. Here, we review the potential therapeutic ability of leptin to correct obesity and other metabolic disorders, not only through its satiating effect, but by also utilizing its thermogenic properties.


Relationship of serum leptin to total and truncal body fat.
1. In this study we investigated the relationship between serum leptin levels and body fat distribution in a random sample of women of widely ranging age and body mass index. Anthropometry and dual energy X-ray absorptiometry were used to measure body fat and its distribution. 2. Leptin levels (log transformed) were not significantly correlated with age, but were significantly positively correlated (P < 0.001) with most anthropometric measures except waist-to-hip circumference ratio. The strongest correlations were with total grams of body fat and percentage body fat (r = 0.68 and 0.76 respectively, P < 0.001). When corrected for percentage body fat the partial correlation coefficients for all other measures became non-significant. The correlation with truncal body fat fell significantly from 0.66 to -0.05 after correction, but the partial correlation with total body fat remained significant (P < 0.005) when grams of truncal fat were controlled for (r = 0.21). 3. These results indicate that the relationship of serum leptin to percentage body fat is the strongest, and that truncal body fat, although the most metabolically active, does not appear to have an independent association with serum leptin.


A Comprehensive Conceptual and Computational Dynamics Framework for Autonomous Regeneration Systems.
Many biological organisms regenerate structure and function after damage. Despite the long history of research on molecular mechanisms, many questions remain about algorithms by which cells can cooperate towards the same invariant morphogenetic outcomes. Therefore, conceptual frameworks are needed not only for motivating hypotheses for advancing the understanding of regeneration processes in living organisms, but also for regenerative medicine and synthetic biology. Inspired by planarian regeneration, this study offers a novel generic conceptual framework that hypothesizes mechanisms and algorithms by which cell collectives may internally represent an anatomical target morphology towards which they build after damage. Further, the framework contributes a novel nature-inspired computing method for self-repair in engineering and robotics. Our framework, based on past in vivo and in silico studies on planaria, hypothesizes efficient novel mechanisms and algorithms to achieve complete and accurate regeneration of a simple in silico flatwormlike organism from any damage, much like the body-wide immortality of planaria, with minimal information and algorithmic complexity. This framework that extends our previous circular tissue repair model integrates two levels of organization: tissue and organism. In Level 1, three individual in silico tissues (head, body, and tail-each with a large number of tissue cells and a single stem cell at the centre) repair themselves through efficient local communications. Here, the contribution extends our circular tissue model to other shapes and invests them with tissue-wide immortality through an information field holding the minimum body plan. In Level 2, individual tissues combine to form a simple organism. Specifically, the three stem cells form a network that coordinates organism-wide regeneration with the help of Level 1. Here we contribute novel concepts for collective decision-making by stem cells for stem cell regeneration and large-scale recovery. Both levels (tissue cells and stem cells) represent networks that perform simple neural computations and form a feedback control system. With simple and limited cellular computations, our framework minimises computation and algorithmic complexity to achieve complete recovery. We report results from computer simulations of the framework to demonstrate its robustness in recovering the organism after any injury. This comprehensive hypothetical framework that significantly extends the existing biological regeneration models offers a new way to conceptualise the information-processing aspects of regeneration, which may also help design living and non-living self-repairing agents.


Extended screened exchange functional derived from transcorrelated density functional theory.
We propose a new formulation of the correlation energy functional derived from the transcorrelated method in use in density functional theory (TC-DFT). An effective Hamiltonian, HTC, is introduced by a similarity transformation of a many-body Hamiltonian, H, with respect to a complex function F: HTC=1FHF. It is proved that an expectation value of HTC for a normalized single Slater determinant, Dn, corresponds to the total energy: E[n] = ⟨Ψn|H|Ψn⟩/⟨Ψn|Ψn⟩ = ⟨Dn|HTC|Dn⟩ under the two assumptions: (1) The electron density nr associated with a trial wave function Ψn = DnF is v-representable and (2) Ψn and Dn give rise to the same electron density nr. This formulation, therefore, provides an alternative expression of the total energy that is useful for the development of novel correlation energy functionals. By substituting a specific function for F, we successfully derived a model correlation energy functional, which resembles the functional form of the screened exchange method. The proposed functional, named the extended screened exchange (ESX) functional, is described within two-body integrals and is parametrized for a numerically exact correlation energy of the homogeneous electron gas. The ESX functional does not contain any ingredients of (semi-)local functionals and thus is totally free from self-interactions. The computational cost for solving the self-consistent-field equation is comparable to that of the Hartree-Fock method. We apply the ESX functional to electronic structure calculations for a solid silicon, H- ion, and small atoms. The results demonstrate that the TC-DFT formulation is promising for the systematic improvement of the correlation energy functional.


Human and preclinical studies of the host-gut microbiome co-metabolite hippurate as a marker and mediator of metabolic health.
Gut microbial products are involved in regulation of host metabolism. In human and experimental studies, we explored the potential role of hippurate, a hepatic phase 2 conjugation product of microbial benzoate, as a marker and mediator of metabolic health. In 271 middle-aged non-diabetic Danish individuals, who were stratified on habitual dietary intake, we applied 1H-nuclear magnetic resonance (NMR) spectroscopy of urine samples and shotgun-sequencing-based metagenomics of the gut microbiome to explore links between the urine level of hippurate, measures of the gut microbiome, dietary fat and markers of metabolic health. In mechanistic experiments with chronic subcutaneous infusion of hippurate to high-fat-diet-fed obese mice, we tested for causality between hippurate and metabolic phenotypes. In the human study, we showed that urine hippurate positively associates with microbial gene richness and functional modules for microbial benzoate biosynthetic pathways, one of which is less prevalent in the Bacteroides 2 enterotype compared with Ruminococcaceae or Prevotella enterotypes. Through dietary stratification, we identify a subset of study participants consuming a diet rich in saturated fat in which urine hippurate concentration, independently of gene richness, accounts for links with metabolic health. In the high-fat-fed mice experiments, we demonstrate causality through chronic infusion of hippurate (20 nmol/day) resulting in improved glucose tolerance and enhanced insulin secretion. Our human and experimental studies show that a high urine hippurate concentration is a general marker of metabolic health, and in the context of obesity induced by high-fat diets, hippurate contributes to metabolic improvements, highlighting its potential as a mediator of metabolic health.


Building a regionally connected reserve network in a changing and uncertain world.
Habitat connectivity is required at large spatial scales to facilitate movement of biota in response to climatic changes and to maintain viable populations of wide-ranging species. Nevertheless, it may require decades to acquire habitat linkages at such scales, and areas that could provide linkages are often developed before they can be reserved. Reserve scheduling methods usually consider only current threats, but threats change over time as development spreads and reaches presently secure areas. We investigated the importance of considering future threats when implementing projects to maintain habitat connectivity at a regional scale. To do so, we compared forward-looking scheduling strategies with strategies that consider only current threats. The strategies were applied to a Costa Rican case study, where many reserves face imminent isolation and other reserves will probably become isolated in the more distant future. We evaluated strategies in terms of two landscape-scale connectivity metrics, a pure connectivity metric and a metric of connected habitat diversity. Those strategies that considered only current threats were unreliable because they often failed to complete planned habitat linkage projects. The most reliable and effective strategies considered the future spread of development and its impact on the likelihood of completing planned habitat linkage projects. Our analyses highlight the critical need to consider future threats when building connected reserve networks over time.


How to detect effects: statistical power and evidence-based practice in occupational therapy research.
The findings from 30 research investigations examining the effectiveness of occupational therapy interventions were reviewed and analyzed. The statistical conclusion validity was determined by computing post hoc power coefficients for the statistical hypothesis tests included in the examined studies. Data analysis revealed the median power values to detect small, medium, and large effect sizes were .09, .33, and .66, respectively. These results suggest a high probability of Type II errors in the sample of occupational therapy intervention research examined. In practical terms, this means the intervention produced a potentially useful treatment effect, but the effect was not detected as significant. Examples are provided that illustrate how low statistical power contributes to increases in Type II errors and inhibits the development of consensus through replication in the research literature. The presence of low-power studies with high rates of false negative findings prevents the establishment of guidelines for evidence-based practice and impedes the scientific progress of rehabilitation professions such as occupational therapy.


Postural balance and fall risk in patients with systemic sclerosis: A cross-sectional study.
This study aims to assess postural balance, fall risk, and the relationship of these parameters with disease-related factors in patients with systemic sclerosis (SSc). Thirty patients with SSc (6 males, 24 females; mean age 51.1±10.6 years; range 35 to 65) and 30 healthy subjects (6 males, 24 females, mean age 52.4±8.7 years; range 35 to 65) matched for age, sex and body mass index were included in this cross-sectional study conducted between September 2018 and November 2019. Postural balance was measured with Biodex Balance SystemTM (Biodex-BS), Berg Balance Scale (BBS), and Timed Up and Go (TUG) test. Individuals' history of falls in the past year, functional capacity, lower limb muscles strength, pulmonary function, respiratory muscle strength, diffusion capacity, and dyspnea severity were evaluated. The SSc group had postural balance impairment and a higher fall frequency than the control group. The SSc group had significantly higher sway index on postural stability (0.6±0.5), lower directional control score (42.1±8.0), and longer test duration (51.8±11.8) on limit of stability of Biodex-BS, lower BBS score (51.5±4.9), and longer test duration on TUG test (8.3±2.7) than control group (all p<0.05). Also, SSc group exhibited significantly lower functional capacity, limb muscles strength, pulmonary function, respiratory muscles strength, diffusion capacity, and higher dyspnea severity than control group (all p<0.05). The postural balance and fall frequency of SSc patients were significantly associated with functional capacity, lower limb muscles strength, pulmonary function, respiratory muscle strength, diffusion capacity, and dyspnea severity. Our results suggest that postural balance impairment and fall risk should be assessed as they appear to be important problems in patients with SSc. Furthermore, assessment of functional capacity, lower limb muscles strength, and lung involvement may highlight those with postural balance impairment and higher fall risk.


Murine Electrophysiological Models of Cardiac Arrhythmogenesis.
Cardiac arrhythmias can follow disruption of the normal cellular electrophysiological processes underlying excitable activity and their tissue propagation as coherent wavefronts from the primary sinoatrial node pacemaker, through the atria, conducting structures and ventricular myocardium. These physiological events are driven by interacting, voltage-dependent, processes of activation, inactivation, and recovery in the ion channels present in cardiomyocyte membranes. Generation and conduction of these events are further modulated by intracellular Ca2+ homeostasis, and metabolic and structural change. This review describes experimental studies on murine models for known clinical arrhythmic conditions in which these mechanisms were modified by genetic, physiological, or pharmacological manipulation. These exemplars yielded molecular, physiological, and structural phenotypes often directly translatable to their corresponding clinical conditions, which could be investigated at the molecular, cellular, tissue, organ, and whole animal levels. Arrhythmogenesis could be explored during normal pacing activity, regular stimulation, following imposed extra-stimuli, or during progressively incremented steady pacing frequencies. Arrhythmic substrate was identified with temporal and spatial functional heterogeneities predisposing to reentrant excitation phenomena. These could arise from abnormalities in cardiac pacing function, tissue electrical connectivity, and cellular excitation and recovery. Triggering events during or following recovery from action potential excitation could thereby lead to sustained arrhythmia. These surface membrane processes were modified by alterations in cellular Ca2+ homeostasis and energetics, as well as cellular and tissue structural change. Study of murine systems thus offers major insights into both our understanding of normal cardiac activity and its propagation, and their relationship to mechanisms generating clinical arrhythmias.


Molecular systematics of the subaerial green algal order Trentepohliales: an assessment based on morphological and molecular data.
The order Trentepohliales is a widespread group of terrestrial green algae. As currently circumscribed, it includes six genera (Cephaleuros, Phycopeltis, Physolinum, Printzina, Stomatochroon and Trentepohlia), the phylogenetic affinities of which are poorly understood. Sequences for the small subunit rRNA gene (18S rDNA) were obtained for several representatives of the order and phylogenetic analyses based on these sequences were compared with the traditional system of classification based on morphological characters. The results indicate that the morphological criteria used for the circumscription of genera and species of the Trentepohliales do not match phylogenetic patterns. Whereas isolates of Cephaleuros form a well-supported monophyletic clade, Trentepohlia is polyphyletic and strains of Trentepohlia, Phycopeltis, Printzina and Physolinum are mixed together in several different lineages. A reassessment of the generic boundaries and a narrower circumscription of some genera (in particular Trentepohlia) will be necessary. Subcuticular habit, heteromorphic life history and occurrence of zoosporangia in clusters are shown to be phylogenetically significant features, whereas other characters commonly used for separation of species and genera (shape and size of the cells, prostrate/erect habit, substratum colonized) have no phylogenetic relevance.


Active stiffening of mitral valve leaflets in the beating heart.
The anterior leaflet of the mitral valve (MV), viewed traditionally as a passive membrane, is shown to be a highly active structure in the beating heart. Two types of leaflet contractile activity are demonstrated: 1) a brief twitch at the beginning of each beat (reflecting contraction of myocytes in the leaflet in communication with and excited by left atrial muscle) that is relaxed by midsystole and whose contractile activity is eliminated with beta-receptor blockade and 2) sustained tone during isovolumic relaxation, insensitive to beta-blockade, but doubled by stimulation of the neurally rich region of aortic-mitral continuity. These findings raise the possibility that these leaflets are neurally controlled tissues, with potentially adaptive capabilities to meet the changing physiological demands on the heart. They also provide a basis for a permanent paradigm shift from one viewing the leaflets as passive flaps to one viewing them as active tissues whose complex function and dysfunction must be taken into account when considering not only therapeutic approaches to MV disease, but even the definitions of MV disease itself.


Social disadvantage, genetic sensitivity, and children's telomere length.
Disadvantaged social environments are associated with adverse health outcomes. This has been attributed, in part, to chronic stress. Telomere length (TL) has been used as a biomarker of chronic stress: TL is shorter in adults in a variety of contexts, including disadvantaged social standing and depression. We use data from 40, 9-y-old boys participating in the Fragile Families and Child Wellbeing Study to extend this observation to African American children. We report that exposure to disadvantaged environments is associated with reduced TL by age 9 y. We document significant associations between low income, low maternal education, unstable family structure, and harsh parenting and TL. These effects were moderated by genetic variants in serotonergic and dopaminergic pathways. Consistent with the differential susceptibility hypothesis, subjects with the highest genetic sensitivity scores had the shortest TL when exposed to disadvantaged social environments and the longest TL when exposed to advantaged environments.


Traffic-related metrics and adverse birth outcomes: A systematic review and meta-analysis.
Given the inconsistency of epidemiologic evidence for associations between maternal exposures to traffic-related metrics and adverse birth outcomes, this manuscript aims to provide clarity on this topic. Pooled meta-estimates were calculated using random-effects analyses. Subgroup analyses were conducted by study area, study design, and Newcastle-Ottawa quality score (NOS). Funnel plots and Egger's test were conducted to evaluate the publication bias, and Fail-safe Numbers (Fail-safe N) were measured to evaluate the robustness of models. From the initial 740 studies (last search, July 11, 2019), 26 studies were included in our analysis. The pooled odds ratio for the change in small for gestational age associated with per 500 m decrease in the distance to roads was 1.016 (95% CI: 1.004, 1.029). Subgroup analyses revealed significant positive associations between term low birth weight and traffic density in higher-quality literatures with higher NOS [1.060 (95% CI: 1.002, 1.121)], cohort studies [1.020 (95% CI: 1.006, 1.033)], and studies in North America [1.018 (95% CI: 1.005, 1.131)]. The buffer of traffic density made no difference in the effect size. Traffic density seemed to be a better indicator of traffic pollution than the distance to roads.


Development of a provisional core set of response measures for clinical trials of systemic sclerosis.
To develop a provisional core set of response measures for clinical trials of systemic sclerosis (SSc). The Scleroderma Clinical Trials Consortium (SCTC) conducted a structured, 3-round Delphi exercise to reach consensus on a core set of measures for clinical trials of SSc. Round 1 asked the SCTC investigators to list items in 11 pre-defined domains (skin, musculoskeletal, cardiac, pulmonary, cardio-pulmonary, gastrointestinal, renal, Raynaud phenomenon and digital ulcers, health-related quality of life and function, global health, and biomarkers) for SSc clinical trials. Round 2 asked respondents to rate the importance of the chosen items and was followed by a meeting, during which the Steering Committee discussed the feasibility, reliability, redundancy and validity of the items. Round 3 sought to obtain broader consensus on the core set measures. Members also voted on items that had data on feasibility but lacked data on reliability and validity, but may still be useful research outcome measures for future trials. A total of 50 SCTC investigators participated in round 1, providing 212 unique items for the 11 domains. In all, 46 (92%) participants responded in round 2 and rated 177 items. The ratings of 177 items were reviewed by the Steering Committee and 31 items from the 11 domains were judged to be appropriate for inclusion in a 1-year multi-centre clinical trial. In total, 40 SCTC investigators completed round 3 and ranked 30 of 31 items as acceptable for inclusion in the core set. The Steering Committee also proposed 14 items for a research agenda. Using a Delphi exercise, we have developed a provisional core set of measures for assessment of disease activity and severity in clinical trials of SSc.


Importance of the long-chain fatty acid beta-hydroxylating cytochrome P450 enzyme YbdT for lipopeptide biosynthesis in Bacillus subtilis strain OKB105.
Bacillus species produce extracellular, surface-active lipopeptides such as surfactin that have wide applications in industry and medicine. The steps involved in the synthesis of 3-hydroxyacyl-coenzyme A (CoA) substrates needed for surfactin biosynthesis are not understood. Cell-free extracts of Bacillus subtilis strain OKB105 synthesized lipopeptide biosurfactants in presence of l-amino acids, myristic acid, coenzyme A, ATP, and H(2)O(2), which suggested that 3-hydroxylation occurs prior to CoA ligation of the long chain fatty acids (LCFAs). We hypothesized that YbdT, a cytochrome P450 enzyme known to beta-hydroxylate LCFAs, functions to form 3-hydroxy fatty acids for lipopeptide biosynthesis. An in-frame mutation of ybdT was constructed and the resulting mutant strain (NHY1) produced predominantly non-hydroxylated lipopeptide with diminished biosurfactant and beta-hemolytic activities. Mass spectrometry showed that 95.6% of the fatty acids in the NHY1 biosurfactant were non-hydroxylated compared to only ∼61% in the OKB105 biosurfactant. Cell-free extracts of the NHY1 synthesized surfactin containing 3-hydroxymyristic acid from 3-hydroxymyristoyl-CoA at a specific activity similar to that of the wild type (17 ± 2 versus 17.4 ± 6 ng biosurfactant min(-1)·ng·protein(-1), respectively). These results showed that the mutation did not affect any function needed to synthesize surfactin once the 3-hydroxyacyl-CoA substrate was formed and that YbdT functions to supply 3-hydroxy fatty acid for surfactin biosynthesis. The fact that YbdT is a peroxidase could explain why biosurfactant production is rarely observed in anaerobically grown Bacillus species. Manipulation of LCFA specificity of YbdT could provide a new route to produce biosurfactants with activities tailored to specific functions.


Comparative study of the protein profiles of Sunki mandarin and Rangpur lime plants in response to water deficit.
Rootstocks play a major role in the tolerance of citrus plants to water deficit by controlling and adjusting the water supply to meet the transpiration demand of the shoots. Alterations in protein abundance in citrus roots are crucial for plant adaptation to water deficit. We performed two-dimensional electrophoresis (2-DE) separation followed by LC/MS/MS to assess the proteome responses of the roots of two citrus rootstocks, Rangpur lime (Citrus limonia Osbeck) and 'Sunki Maravilha' (Citrus sunki) mandarin, which show contrasting tolerances to water deficits at the physiological and molecular levels. Changes in the abundance of 36 and 38 proteins in Rangpur lime and 'Sunki Maravilha' mandarin, respectively, were observed via LC/MS/MS in response to water deficit. Multivariate principal component analysis (PCA) of the data revealed major changes in the protein profile of 'Sunki Maravilha' in response to water deficit. Additionally, proteomics and systems biology analyses allowed for the general elucidation of the major mechanisms associated with the differential responses to water deficit of both varieties. The defense mechanisms of Rangpur lime included changes in the metabolism of carbohydrates and amino acids as well as in the activation of reactive oxygen species (ROS) detoxification and in the levels of proteins involved in water stress defense. In contrast, the adaptation of 'Sunki Maravilha' to stress was aided by the activation of DNA repair and processing proteins. Our study reveals that the levels of a number of proteins involved in various cellular pathways are affected during water deficit in the roots of citrus plants. The results show that acclimatization to water deficit involves specific responses in Rangpur lime and 'Sunki Maravilha' mandarin. This study provides insights into the effects of drought on the abundance of proteins in the roots of two varieties of citrus rootstocks. In addition, this work allows for a better understanding of the molecular basis of the response to water deficit in citrus. Further analysis is needed to elucidate the behaviors of the key target proteins involved in this response.


Long-term multi-species Lactobacillus and Bifidobacterium dietary supplement enhances memory and changes regional brain metabolites in middle-aged rats.
Ageing is associated with changes in the gut microbiome that may contribute to age-related changes in cognition. Previous work has shown that dietary supplements with multi-species live microorganisms can influence brain function, including induction of hippocampal synaptic plasticity and production of brain derived neurotrophic factor, in both young and aged rodents. However, the effect of such dietary supplements on memory processes has been less well documented, particularly in the context of aging. The main aim of the present study was to examine the impact of a long-term dietary supplement with a multi-species live Lactobacillus and Bifidobacteria mixture (Lactobacillus acidophilus CUL60, L. acidophilus CUL21, Bifidobacterium bifidum CUL20 and B. lactis CUL34) on tests of memory and behavioural flexibility in 15-17-month-old male rats. Following behavioural testing, the hippocampus and prefrontal cortex was extracted and analysed ex vivo using 1H nuclear magnetic resonance (1H NMR) spectroscopy to examine brain metabolites. The results showed a small beneficial effect of the dietary supplement on watermaze spatial navigation and robust improvements in long-term object recognition memory and short-term memory for object-in-place associations. Short-term object novelty and object temporal order memory was not influenced by the dietary supplement in aging rats. 1H NMR analysis revealed diet-related regional-specific changes in brain metabolites; which indicated changes in several pathways contributing to modulation of neural signaling. These data suggest that chronic dietary supplement with multi-species live microorganisms can alter brain metabolites in aging rats and have beneficial effects on memory.


The long-term safety of danazol in women with hereditary angioedema.
Although the short-term safety (less than or equal to 6 months) of danazol has been established in a variety of settings, no information exists as to its long-term safety. We therefore investigated the long-term safety of danazol by performing a retrospective chart review of 60 female patients with hereditary angioedema treated with danazol for a continuous period of 6 months or longer. The mean age of the patients was 35.2 years and the mean duration of therapy was 59.7 months. Virtually all patients experienced one or more adverse reactions. Menstrual abnormalities (79%), weight gain (60%), muscle cramps/myalgias (40%), and transaminase elevations (40%) were the most common adverse reactions. The drug was discontinued due to adverse reactions in 8 patients. No patient has died or suffered any apparent long-term sequelae that were directly attributable to the drug. We conclude that, despite a relatively high incidence of adverse reactions, danazol has proven to be remarkably safe over the long-term in this group of patients.


Staphylococcal Enterotoxin B-specific monoclonal antibody 20B1 successfully treats diverse Staphylococcus aureus infections.
Methicillin-resistant Staphylococcus aureus (MRSA) has become a major health threat in the United States. Staphylococcal enterotoxin B (SEB) is a potent superantigen that contributes to its virulence. High mortality and frequent failure of therapy despite available antibiotics have stimulated research efforts to develop adjunctive therapies. Treatment benefits of SEB-specific monoclonal antibody (mAb) 20B1 were investigated in mice in sepsis, superficial skin, and deep-tissue infection models. Mice challenged with a SEB-producing MRSA strain developed fatal sepsis, extensive tissue skin infection, and abscess-forming deep-seeded thigh muscle infection. Animals preimmunized against SEB or treated passively with mAb 20B1 exhibited enhanced survival in the sepsis model, whereas decrease of bacterial burden was observed in the superficial skin and deep-tissue models. mAb 20B1 bound to SEB in the infected tissue and decreased abscess formation and proinflammatory cytokine levels, lymphocyte proliferation, and neutrophil recruitment. mAb 20B1, an SEB-neutralizing mAb, is effective against MRSA infection. mAb 20B1 protects against lethal sepsis and reduces skin tissue invasion and deep-abscess formation. The mAb penetrates well into the abscess and binds to SEB. It affects the outcome of S. aureus infection by modulating the host's proinflammatory immune response.


A role for calcium-calmodulin in regulating nitric oxide production during skeletal muscle satellite cell activation.
When skeletal muscle is stretched or injured, myogenic satellite cells are activated to enter the cell cycle. This process depends on nitric oxide (NO) production by NO synthase (NOS), matrix metalloproteinase activation, release of hepatocyte growth factor (HGF) from the extracellular matrix, and presentation of HGF to the c-met receptor as demonstrated by a primary culture and in vivo assays. We now add evidence that calcium-calmodulin is involved in the satellite cell activation cascade in vitro. Conditioned medium from cultures that were treated with a calcium ionophore (A23187, ionomycin) for 2 h activated cultured satellite cells and contained active HGF, similar to the effect of mechanical stretch or NO donor treatments. The response was abolished by addition of calmodulin inhibitors (calmidazolium, W-13, W-12) or a NOS inhibitor N(G)-nitro-l-arginine methyl ester hydrochloride but not by its less inactive enantiomer N(G)-nitro-d-arginine methyl ester hydrochloride. Satellite cells were also shown to express functional calmodulin protein having a calcium-binding activity at 12 h postplating, which is the time at which the calcium ionophore was added in this study and the stretch treatment was applied in our previous experiments. Therefore, results from these experiments provide an additional insight that calcium-calmodulin mediates HGF release from the matrix and that this step in the activation pathway is upstream from NO synthesis.


The polyol process: a unique method for easy access to metal nanoparticles with tailored sizes, shapes and compositions.
After about three decades of development, the polyol process is now widely recognized and practised as a unique soft chemical method for the preparation of a large variety of nanoparticles which can be used in important technological fields. It offers many advantages: low cost, ease of use and, very importantly, already proven scalability for industrial applications. Among the different classes of inorganic nanoparticles which can be prepared in liquid polyols, metals were the first reported. This review aims to give a comprehensive account of the strategies used to prepare monometallic nanoparticles and multimetallic materials with tailored size and shape. As regards monometallic materials, while the preparation of noble as well as ferromagnetic metals is now clearly established, the scope of the polyol process has been extended to the preparation of more electropositive metals, such as post-transition metals and semi-metals. The potential of this method is also clearly displayed for the preparation of alloys, intermetallics and core-shell nanostructures with a very large diversity of compositions and architectures.


Magnitude Squared Coherence Method based on Weighted Canonical Correlation Analysis for EEG Synchronization Analysis in Amnesic Mild Cognitive Impairment of Diabetes Mellitus.
Type 2 diabetes mellitus (T2DM) increases the risk of amnestic mild cognitive impairment (aMCI) and Alzheimer's disease (AD). aMCI is the transitory stage from normal cognition to AD, which seriously impacts the quality of human life, especially for old people. Electroencephalography (EEG) coherence can assess the functional connectivity between different brain regions, which is an effective way to research the pathogenesis of aMCI and distinguish aMCI patients from normal cognitive subjects. In this paper, we propose a new EEG coherence approach named magnitude squared coherence based on weighted canonical correlation analysis (WCCA-MSC) which improves the accuracy in coherence estimation by the means of weighting. In comparison to the magnitude squared coherence based on canonical correlation analysis (CCA-MSC) and magnitude squared coherence based on reduced-rank canonical correlation analysis (RCCA-MSC), the proposed WCCA-MSC behaves better in the influence of noise and relative amplitude of frequency components and provides more accuracy at non-coherent frequencies. The application of the new method to EEG signals showed increased coherence in Delta and Theta frequency bands decreased coherence in an Alpha frequency band in aMCI patients. Considering its property of better performance, the proposed coherence method is of great advantage in analyzing the pathogenesis of aMCI with T2DM.


Fas ligand-mediated bystander lysis of syngeneic cells in response to an allogeneic stimulus.
Using a mouse allospecific lymph node (LN) CTL (H-2b) response to transplanted cells (H-2k), effector cells were shown to lyse C3H/HeJ or L929-Fas (H-2k) target cells in either a perforin-dependent or a Fas ligand (FasL)-dependent manner. C3H/HeJ.lpr targets or L929 targets lacking Fas were not sensitive to these effectors generated in perforin-deficient (Po) mice. By contrast, C3H/HeJ.lpr target cells and L929 were as responsive as C3H/HeJ and L929-Fas targets to the effectors generated in perforin wild-type (P+/+) mice. To measure potential bystander lysis in allogeneic responses, these same effector LN from either C57BL/6 P+/+ or P(o) mice were examined for lysis of C3H/HeJ (or C3H/HeJ.lpr) or L929-Fas (or L929) cocultured with 51Cr-labeled C57BL/6 (H-2b) target cells. Bystander lysis of C57BL/6 targets was observed only in those allogeneic responses where FasL was the defined mechanism of lysis. By contrast, perforin-mediated lysis was restricted by allo-recognition. Syngeneic (H-2b) targets that were not sensitive to Fas-mediated lysis were not lysed in a bystander fashion. Depletion of subsets of LN Po effectors (b anti-k) demonstrated that CD4+ T cells were primarily responsible for FasL-mediated direct and bystander lysis. FasL-mediated bystander lysis was also observed in H-2k anti-b reactions, except when the effectors were FasL mutant (gld). These data suggest that CTL responses to foreign Ag may evoke a certain amount of collateral damage to local host Fas-sensitive cells.


Odor localization requires visual feedback during free flight in Drosophila melanogaster.
Adult fruit flies follow attractive odors associated with food and oviposition sites through widely varied visual landscapes. To examine the interaction between olfactory and visual cues during search behavior, we recorded three-dimensional flight trajectories as individuals explored controlled sensory landscapes. When presented with the source of an attractive odor invisibly embedded in the floor of a 1 m arena, flies spend most of their time hovering back and forth over the source when flying within a randomly textured visual background but fail to localize the source when searching within a uniform white surround. To test whether flies are associating unique features of the visual background with the strength of odor cues, we flew them within arenas containing evenly spaced vertical stripes. Flies readily localized the odor when flying within visual landscapes lacking azimuthal landmarks provided that vertical edges were present. Flies failed to localize odor when flying within a background pattern consisting of horizontal stripes. These results suggest that, whereas flies do not require spatially unique visual patterns to localize an odor source, they do require visual feedback generated by vertical edges. Quantitative shifts in several components of flight behavior accompanied successful odor localization. Flies decrease flight altitude, turn more often and approach visually textured walls of the arena near an odor source. A simple model based on the statistics of flight behavior supports the hypothesis that a subtle influence on these behaviors is sufficient to lead a fly to its food.


Packing morphology of wavy nanofiber arrays.
Existing theories for quantifying the morphology of nanofibers (NFs) in aligned arrays either neglect or assume a simple functional form for the curvature of the NFs, commonly known as the NF waviness. However, since such assumptions cannot adequately describe the waviness of real NFs, errors that can exceed 10% in the predicted inter-NF separation can result. Here we use a theoretical framework capable of simulating >10(5) NFs with stochastic three-dimensional morphologies to quantify NF waviness on an easily accessible measure of the morphology, the inter-NF spacing, for a range of NF volume fractions. The presented scaling of inter-NF spacing with waviness is then used to study the morphology evolution of aligned carbon nanotube (A-CNT) arrays during packing, showing that the effective two-dimensional coordination number of the A-CNTs increases much faster than previously reported during close packing, and that hexagonal close packing can successfully describe the packing morphology of the A-CNTs at volume fractions greater than 40 vol%.


Transport of 3-bromopyruvate across the human erythrocyte membrane.
3-Bromopyruvic acid (3-BP) is a promising anticancer compound because it is a strong inhibitor of glycolytic enzymes, especially glyceraldehyde 3-phosphate dehydrogenase. The Warburg effect means that malignant cells are much more dependent on glycolysis than normal cells. Potential complications of anticancer therapy with 3-BP are side effects due to its interaction with normal cells, especially erythrocytes. Transport into cells is critical for 3-BP to have intracellular effects. The aim of our study was the kinetic characterization of 3-BP transport into human erythrocytes. 3-BP uptake by erythrocytes was linear within the first 3 min and pH-dependent. The transport rate decreased with increasing pH in the range of 6.0-8.0. The Km and Vm values for 3-BP transport were 0.89 mM and 0.94 mmol/(l cells x min), respectively. The transport was inhibited competitively by pyruvate and significantly inhibited by DIDS, SITS, and 1-cyano-4-hydroxycinnamic acid. Flavonoids also inhibited 3-BP transport: the most potent inhibition was found for luteolin and quercetin.


Working Memory Load-related Theta Power Decreases in Dorsolateral Prefrontal Cortex Predict Individual Differences in Performance.
Holding information in working memory (WM) is an active and effortful process that is accompanied by sustained load-dependent changes in oscillatory brain activity. These proportional power increases are often reported in EEG studies recording theta over frontal midline sites. Intracranial recordings, however, yield mixed results, depending on the brain area being recorded from. We recorded intracranial EEG with depth electrodes in 13 patients with epilepsy who were performing a Sternberg WM task. Here, we investigated patterns of theta power changes as a function of memory load during maintenance in three areas critical for WM: dorsolateral prefrontal cortex (DLPFC), dorsal ACC (dACC), and hippocampus. Theta frequency power in both hippocampus and dACC increased during maintenance. In contrast, theta frequency power in the DLPFC decreased during maintenance, and this decrease was proportional to memory load. Only the power decreases in DLPFC, but not the power increases in hippocampus and dACC, were predictive of behavior in a given trial. The extent of the load-related theta power decreases in the DLPFC in a given participant predicted a participant's RTs, revealing that DLPFC theta explains individual differences in WM ability between participants. Together, these data reveal a pattern of theta power decreases in the DLPFC that is predictive of behavior and that is opposite of that in other brain areas. This result suggests that theta band power changes serve different cognitive functions in different brain areas and specifically that theta power decreases in DLPFC have an important role in maintenance of information.


Turning Attention Inside Out: How Working Memory Serves Behavior.
Flexible behavior requires guidance not only by sensations that are available immediately but also by relevant mental contents carried forward through working memory. Therefore, selective-attention functions that modulate the contents of working memory to guide behavior (inside-out) are just as important as those operating on sensory signals to generate internal contents (outside-in). We review the burgeoning literature on selective attention in the inside-out direction and underscore its functional, flexible, and future-focused nature. We discuss in turn the purpose (why), targets (what), sources (when), and mechanisms (how) of selective attention inside working memory, using visual working memory as a model. We show how the study of internal selective attention brings new insights concerning the core cognitive processes of attention and working memory and how considering selective attention and working memory together paves the way for a rich and integrated understanding of how mind serves behavior.


Percutaneous techniques for the diagnosis and treatment of localized Langerhans-cell histiocytosis (eosinophilic granuloma of bone).
We retrospectively studied the outcome of percutaneous needle biopsy and intralesional injection of a corticosteroid (methylprednisolone) in thirty-nine patients who had localized Langerhans-cell histiocytosis (eosinophilic granuloma of bone). All thirty-nine patients had a solitary symptomatic lesion at presentation; a second lesion developed in two patients, two and four months after the first lesion was diagnosed. Therefore, there were forty-one lesions in thirty-nine patients. Fine-needle aspiration with or without core-needle biopsy was performed for all forty-one lesions, and the diagnosis of Langerhans-cell histiocytosis was established for thirty-seven (90 per cent). A corticosteroid was injected into thirty-five lesions. Twenty-nine received the injection at the time of the fine-needle aspiration on the basis of the cytological findings in the aspirate. Six patients who had a solitary lesion had a two-stage procedure because the injection was delayed until the diagnosis was confirmed with histological evaluation of specimens obtained by core-needle biopsy. Thirty-four (97 per cent) of the thirty-five lesions healed. The clinical symptoms associated with thirty-one lesions resolved within two weeks after a single injection of the corticosteroid. There were no complications associated with either the biopsy or the injection. At a median of ninety months (range, twenty-four to 199 months), no patient had recurrence of symptoms or of radiographic evidence of the lesion. All patients who had been managed with an intralesional injection of the corticosteroid had full range of motion of the affected extremity and had resumed unlimited activities. Although the mechanism of action of intralesional injection of a corticosteroid has not been defined, use of percutaneous needle biopsy to diagnose localized Langerhans-cell histiocytosis and treatment with intralesional administration of methylprednisolone relieved pain expeditiously, enabled the patient to avoid an operative procedure, and resulted in osseous healing. The specific role of corticosteroid therapy remains to be determined by prospective, randomized studies.


blaI and blaR1 regulate beta-lactamase and PBP 2a production in methicillin-resistant Staphylococcus aureus.
For Staphylococcus aureus, it is hypothesized that two genes located upstream of the beta-lactamase gene, blaZ, are required for the inducible expression of beta-lactamase. blaR1 is predicted to encode a signal-transducing membrane protein, and blaI is predicted to encode a repressor protein. These same two genes may also regulate the production of penicillin-binding protein 2a (PBP 2a), a protein essential for expression of methicillin resistance. To confirm that these two genes encode products that can control both beta-lactamase and PBP 2a production, blaI, blaR1, and blaZ with a 150-nucleotide deletion at the 3' end were subcloned from a 30-kb staphylococcal beta-lactamase plasmid and three beta-lactamase-negative strains of methicillin-resistant S. aureus were transformed with the recombinant plasmid containing that insert. The production of PBP 2a and a nonfunctional beta-lactamase was detected by fluorography and by immunoblots with polyclonal antisera directed against each of the proteins. Whereas the parent strains did not produce beta-lactamase and constitutively produced PBP 2a, PBP 2a and a truncated beta-lactamase were now inducible in the transformants. Therefore, two plasmid-derived genes regulate the production of both PBP 2a and beta-lactamase.


A USA registry of gastrointestinal stromal tumor patients: changes in practice over time and differences between community and academic practices.
The objective of the study was to describe patterns of care of patients with gastrointestinal stromal tumors (GISTs) in the United States in the tyrosine kinase inhibitor (TKI) era. From November 2004 through March 2009, data were collected regarding demographics, diagnostic history, treatment, relapse, and survival of 882 patients with GIST from 122 community and academic medical practices. The most common first-line treatment for the 719 patients presenting with localized GIST was surgery (87%). Use of adjuvant imatinib increased after June 2007; 47% of patients enrolled in the registry considered by the investigator to be at high risk for recurrence received adjuvant imatinib after June 2007 versus 18% before. Overall, 56% of patients received imatinib and 11% received sunitinib. The utilization of targeted therapy increased over time (45% and 0.4% of patients received imatinib and sunitinib, respectively, in 2006 versus 56% and 11%, respectively, in 2009). These are the first GIST registry data from the TKI era. The use of targeted therapy for GIST has increased in accordance with updated treatment guidelines. Diagnosis of GIST has evolved with increased use of KIT testing. The duration of targeted therapy in the adjuvant therapy setting is similar in community and academic practices.


Mitochondrial pathogenic mutations are population-specific.
Surveying deleterious variation in human populations is crucial for our understanding, diagnosis and potential treatment of human genetic pathologies. A number of recent genome-wide analyses focused on the prevalence of segregating deleterious alleles in the nuclear genome. However, such studies have not been conducted for the mitochondrial genome. We present a systematic survey of polymorphisms in the human mitochondrial genome, including those predicted to be deleterious and those that correspond to known pathogenic mutations. Analyzing 4458 completely sequenced mitochondrial genomes we characterize the genetic diversity of different types of single nucleotide polymorphisms (SNPs) in African (L haplotypes) and non-African (M and N haplotypes) populations. We find that the overall level of polymorphism is higher in the mitochondrial compared to the nuclear genome, although the mitochondrial genome appears to be under stronger selection as indicated by proportionally fewer nonsynonymous than synonymous substitutions. The African mitochondrial genomes show higher heterozygosity, a greater number of polymorphic sites and higher frequencies of polymorphisms for synonymous, benign and damaging polymorphism than non-African genomes. However, African genomes carry significantly fewer SNPs that have been previously characterized as pathogenic compared to non-African genomes. Finding SNPs classified as pathogenic to be the only category of polymorphisms that are more abundant in non-African genomes is best explained by a systematic ascertainment bias that favours the discovery of pathogenic polymorphisms segregating in non-African populations. This further suggests that, contrary to the common disease-common variant hypothesis, pathogenic mutations are largely population-specific and different SNPs may be associated with the same disease in different populations. Therefore, to obtain a comprehensive picture of the deleterious variability in the human population, as well as to improve the diagnostics of individuals carrying African mitochondrial haplotypes, it is necessary to survey different populations independently. This article was reviewed by Dr Mikhail Gelfand, Dr Vasily Ramensky (nominated by Dr Eugene Koonin) and Dr David Rand (nominated by Dr Laurence Hurst).


Effects of Long Chain Omega-3 Polyunsaturated Fatty Acids on Brain Function in Mildly Hypertensive Older Adults.
Purported benefits of long chain omega-3 polyunsaturated fatty acid (LCn-3PUFA) for brain function may be attributable, at least in part, to improved cerebral perfusion. A pilot randomised controlled trial was undertaken to investigate effects of taking a DHA-rich fish oil supplement for 20 weeks on cerebrovascular function, mood and cognitive performance. Borderline hypertensives aged 40⁻85 years with low habitual LCn-3PUFA intake took four capsules/day of EPAX (1600 mg DHA + 400 mg EPA) or placebo (corn oil). Cerebrovascular function was assessed at baseline and after 20 weeks in 38 completers (19 on each supplement) using transcranial Doppler ultrasound of blood flow in the middle cerebral artery at rest and whilst performing a battery of cognitive tasks (neurovascular coupling). The primary outcome, cerebrovascular responsiveness (CVR) to hypercapnia, increased 26% (p = 0.024) in women; there was no change in men. In contrast, neurovascular coupling increased significantly (p = 0.01 for the overall response) in men only; the latter correlated with an increase of EPA in erythrocytes (r = 0.616, p = 0.002). There was no associated improvement of mood or cognition in either men or women. These preliminary observations indicate that LCn-3PUFA supplementation has the potential to enhance blood flow in the brain in response to both hypercapnic and cognitive stimuli. Future studies should examine differential effects of EPA and DHA and take account of the gender differences in responsiveness to supplementation.


Selective and directional actuation of elastomer films using chained magnetic nanoparticles.
We report selective and directional actuation of elastomer films utilizing magnetic anisotropy introduced by chains of Fe3O4 magnetic nanoparticles (MNPs). Under uniform magnetic fields or field gradients, dipolar interactions between the MNPs favor magnetization along the chain direction and cause selective lifting. This mechanism is described using a simple model.


Contraction kinetics of red muscle in scup: mechanism for variation in relaxation rate along the length of the fish.
We studied possible mechanisms for the twofold difference in red muscle relaxation times between the posterior (207.2 ms) and anterior (98.4 ms) musculature of scup Stenotomus chrysops, which has been shown to have a large effect on power generation during swimming. This difference was not due to contamination of the anterior bundles with faster fiber types, as histological examination showed that all bundles contained more than 98.9% red fibers. Further, maximum velocities of shortening (Vmax) at 20 degrees C were nearly identical, 5.37 MLs-1 (where ML is muscle length) for the anterior musculature and 5.47 MLs-1 for the posterior musculature, suggesting that the difference in relaxation times was not due to a difference in the crossbridge detachment rates associated with different myosin isoforms. The possibility of differences in the Ca2+ pumping rate influencing relaxation rate was explored using cyclopiazonic acid (CPA), a sarcoplasmic reticulum (SR) Ca(2+)-ATPase inhibitor. The concentration of CPA could be adjusted to slow the relaxation rate of an anterior muscle to that of a posterior muscle. However, SDS gels showed no difference in the intensity of SR Ca(2+)-ATPase protein bands between muscle positions. These results suggest that differences in the Ca2+ pumping could account for the observed difference in relaxation rate, but do not support the simplest hypothesis that the difference in relaxation rates is due to differences in numbers of Ca2+ pumps. Other possible mechanisms for this difference are explored.


Predicting classifier performance with limited training data: applications to computer-aided diagnosis in breast and prostate cancer.
Clinical trials increasingly employ medical imaging data in conjunction with supervised classifiers, where the latter require large amounts of training data to accurately model the system. Yet, a classifier selected at the start of the trial based on smaller and more accessible datasets may yield inaccurate and unstable classification performance. In this paper, we aim to address two common concerns in classifier selection for clinical trials: (1) predicting expected classifier performance for large datasets based on error rates calculated from smaller datasets and (2) the selection of appropriate classifiers based on expected performance for larger datasets. We present a framework for comparative evaluation of classifiers using only limited amounts of training data by using random repeated sampling (RRS) in conjunction with a cross-validation sampling strategy. Extrapolated error rates are subsequently validated via comparison with leave-one-out cross-validation performed on a larger dataset. The ability to predict error rates as dataset size increases is demonstrated on both synthetic data as well as three different computational imaging tasks: detecting cancerous image regions in prostate histopathology, differentiating high and low grade cancer in breast histopathology, and detecting cancerous metavoxels in prostate magnetic resonance spectroscopy. For each task, the relationships between 3 distinct classifiers (k-nearest neighbor, naive Bayes, Support Vector Machine) are explored. Further quantitative evaluation in terms of interquartile range (IQR) suggests that our approach consistently yields error rates with lower variability (mean IQRs of 0.0070, 0.0127, and 0.0140) than a traditional RRS approach (mean IQRs of 0.0297, 0.0779, and 0.305) that does not employ cross-validation sampling for all three datasets.


Multiple personality disorder: an overview.
Multiple personality disorder is understood today as chronic dissociative psychopathology that most often develops in response to severe abuse in childhood. The dissociative component is a manifestation of a defense mechanism out of control. The person with a biopsychological capacity to dissociate flees inward from overwhelming abuse or feared abuse. When continuing abuse perpetuates dissociations and they are chained by common affective themes, the foundations of multiple personality disorder are laid. Although the disorder has its roots in childhood, most patients are not diagnosed with this condition until 20 to 50 years of age. Many have received several prior, erroneous diagnoses of mental or physical disorders or both over a period of 7 or more years. Failure of diagnosis is an indication of the multiple factors that contribute to making this a covert disorder. Diagnosis and management begin at the same place: the establishment of trust and therapeutic alliance between patient and therapist.


NMDA receptor subtypes at autaptic synapses of cerebellar granule neurons.
We studied the action potential-evoked autaptic N-methyl-d-aspartate receptor-mediated excitatory postsynaptic currents (NMDA-EPSCs) using solitary cerebellar neurons cultured in microislands from wild-type (+/+), NR2A subunit knockout (NR2A-/-), and NR2C subunit knockout (NR2C-/-) mice. The peak amplitude of autaptic NMDA-EPSCs increased for all genotypes between days in vitro 8 (DIV8) and DIV13. Compared with +/+ cells at DIV13, NR2A-/- cells had smaller and NR2C-/- cells had larger NMDA-EPSCs. The decay time of these currents were all unexpectedly fast, except in NR2A-/- neurons, and showed small but significant shortening with development. Comparison of quantal parameters during development indicated an increase in quantal content in all genotypes. The synaptic portion of NMDA receptors measured using MK-801 blockade was roughly 50% in all genotypes at DIV8, and this percentage became slightly larger in NR2A-/- and NR2C-/- neurons at DIV12. The NR2B-selective antagonists Conantokin G and CP101,606 differed in their blocking actions with development, suggesting the presence of both heterodimeric NR1/NR2B and heterotrimeric NR1/NR2A/NR2B receptors. The most striking result we obtained was the significant increase of NMDA-EPSC peak amplitude and charge transfer in NR2C-/- mice. This was mainly the result of an increase in quantal size as estimated from miniature NMDA-EPSCs. The expression of NR2C subunit containing receptors was supported by the decreased Mg(2+) sensitivity of NMDA receptors at DIV13 in +/+ but not in NR2C-/- cells. Thus solitary cerebellar granule neurons provide a novel model to investigate the role of receptor subtypes in the developmental changes of synaptic NMDA receptors.


The developmental proteome of Drosophila melanogaster.
Drosophila melanogaster is a widely used genetic model organism in developmental biology. While this model organism has been intensively studied at the RNA level, a comprehensive proteomic study covering the complete life cycle is still missing. Here, we apply label-free quantitative proteomics to explore proteome remodeling across Drosophila's life cycle, resulting in 7952 proteins, and provide a high temporal-resolved embryogenesis proteome of 5458 proteins. Our proteome data enabled us to monitor isoform-specific expression of 34 genes during development, to identify the pseudogene Cyp9f3Ψ as a protein-coding gene, and to obtain evidence of 268 small proteins. Moreover, the comparison with available transcriptomic data uncovered examples of poor correlation between mRNA and protein, underscoring the importance of proteomics to study developmental progression. Data integration of our embryogenesis proteome with tissue-specific data revealed spatial and temporal information for further functional studies of yet uncharacterized proteins. Overall, our high resolution proteomes provide a powerful resource and can be explored in detail in our interactive web interface.


Systems initiatives reduce healthcare-associated infections: a study of 22,928 device days in a single trauma unit.
"Implementation research" promotes the systematic conversion of evidence-based principles into routine practice to improve the quality of care. We hypothesized a system-based initiative to reduce nosocomial infection would lower the incidence of ventilator-associated pneumonia (VAP), urinary tract infection (UTI), and bloodstream infection (BSI). From January 2006 to April 2008, 7,364 adult trauma patients were admitted, of which 1,953 (27%) were admitted to the trauma intensive care unit and comprised the study group. Tight glycemic control was maintained using a computer algorithm for continuous insulin administration based on every 2-hour blood glucose testing. Centers for Disease Control and Prevention definitions of nosocomial infections were used. Evidence-based infection reduction strategies included the following: a VAP bundle (spontaneous breathing, Richmond Agitation-Sedation Scale, oral hygiene, bed elevation, and deep vein thrombosis/stress ulcer prophylaxis), UTI (expert insertion team and Foley removal/change at 5 days), and BSI (maximum barrier precautions, chlorhexidine skin prep, line management protocol). An electronic dashboard identified the at-risk population, and designated auditors monitored the compliance. Infection rates (events per 1,000 device days) were measured over time and compared annually using Fisher's exact test. The study group had 22,928 device exposure days: 6,482 ventilator days, 9,037 urinary catheter days, and 7,399 central line days. Patient acuity, demographics, and number of device days did not vary significantly year-to-year. Annual infection rates declined between 2006 and 2008, and decreases in UTI and BSI rates were statistically significant (p < 0.05). These decreases pushed UTI and BSI rates below Centers for Disease Control and Prevention norms. Over 28 months, a systems approach to reducing nosocomial infection rates after trauma decreased nosocomial infections: UTI (76.3%), BSI (74.1%), and VAP (24.9%). Our experience suggests that infection reduction requires (1) an evidence-based plan; (2) MD and staff education/commitment; (3) electronic documentation; and (4) auditors to monitor and ensure compliance.


Complications of plantar fascia rupture associated with corticosteroid injection.
From 1992 to 1995, 765 patients with a clinical diagnosis of plantar fasciitis were evaluated by one of the authors. Fifty-one patients were diagnosed with plantar fascia rupture, and 44 of these ruptures were associated with corticosteroid injection. The authors injected 122 of the 765 patients, resulting in 12 of the 44 plantar fascia ruptures. Subjective and objective evaluations were conducted through chart and radiographic review. Thirty-nine of these patients were evaluated at an average 27-month follow-up. Thirty patients (68%) reported a sudden onset of tearing at the heel, and 14 (32%) had a gradual onset of symptoms. In most cases the original heel pain was relieved by rupture. However, these patients subsequently developed new problems including longitudinal arch strain, lateral and dorsal midfoot strain, lateral plantar nerve dysfunction, stress fracture, hammertoe deformity, swelling, and/or antalgia. All patients exhibited diminished tension of the plantar fascia upon examination by the stretch test. Comparison of calcaneal pitch angles in the affected and uninvolved foot showed a statistically significant difference of 3.7 degrees (P = 0.0001). Treatment included NSAIDs, rest or cross-training, stretching, orthotics, and boot-brace immobilization. At an average 27-month follow-up, 50% had good/excellent scores and 50% had fair/poor scores. Recovery time was varied. Ten feet were asymptomatic by 6 months post rupture, four feet by 12 months post rupture, and 26 feet remained symptomatic 1 year post rupture. Our findings demonstrate that plantar fascia rupture after corticosteroid injection may result in long-term sequelae that are difficult to resolve.


A dip-stick type biosensor using bioluminescent bacteria encapsulated in color-coded alginate microbeads for detection of water toxicity.
The use of genetically engineered bioluminescent bacteria, in which bioluminescence is induced by different modes of toxic action, represents an alternative to acute toxicity tests using living aquatic organisms (plants, vertebrates, or invertebrates) in an aqueous environment. A number of these bacterial strains have been developed, but there have been no attempts to develop a hand-held type of biosensor for monitoring or identification of toxicity. We report a facile dip-stick type biosensor using genetically engineered bioluminescent bacteria as a new platform for classification and identification of toxicity in water environments. This dip-stick type biosensor is composed of eight different optically color-coded functional alginate beads that each encapsulates a different bioluminescent bacterial strain and its corresponding fluorescent microbead. These color-coded microbeads exhibit easy identification of encapsulated microbeads, since each microbead has a different color code depending on the bioluminescent bacterial strain contained and improved cell-stability compared to liquid culture. This dip-stick type biosensor can discriminate different modes of toxic actions (i.e. DNA damage, oxidative damage, cell-membrane damage, or protein damage) of sample water tested by simply dipping the stick into the water samples. It was found that each color-coded microbead emitted distinct bioluminescence, and each dip-stick type biosensor showed different bioluminescence patterns within 2 hours, depending on the toxic chemicals contained in LB medium, tap water, or river water samples. This dip-stick type biosensor can, therefore, be widely and practically used in checking toxicity of water in the environment primarily in situ, possibly indicating the status of biodiversity.


Combining Explicit Quantum Solvent with a Polarizable Continuum Model.
A promising approach for accurately modeling both short-range and long-range solvation effects is to combine explicit quantum mechanical (QM) solvent with a classical polarizable continuum model (PCM), but the best PCM for these combined QM/classical calculations is relatively unexplored. We find that the choice of the solvation cavity is very important for obtaining physically correct results since unphysical double counting of solvation effects from both the QM solvent and the classical dielectric can occur with a poor choice of cavity. We investigate the dependence of electronic excitation energies on the definition of the PCM cavity and the self-consistent reaction field method, comparing results to large-scale explicit QM solvent calculations. For excitation energies, we identify the difference between the ground and excited state dipole moments as the key property determining the sensitivity to the PCM cavity. Using a linear response PCM approach combined with QM solvent, we show that excitation energies are best modeled by a solvent excluded surface or a scaled van der Waals surface. For the aqueous solutes studied here, we find that a scaled van der Waals surface defined by universal force field radii scaled by a factor of 1.5 gives reasonable excitation energies. When using an external iteration state-specific PCM approach, however, the excitation energies are most accurate with a larger PCM cavity, such as a solvent accessible surface.


From sulfur to homoglutathione: thiol metabolism in soybean.
Sulfur is an essential plant nutrient and is metabolized into the sulfur-containing amino acids (cysteine and methionine) and into molecules that protect plants against oxidative and environmental stresses. Although studies of thiol metabolism in the model plant Arabidopsis thaliana (thale cress) have expanded our understanding of these dynamic processes, our knowledge of how sulfur is assimilated and metabolized in crop plants, such as soybean (Glycine max), remains limited in comparison. Soybean is a major crop used worldwide for food and animal feed. Although soybeans are protein-rich, they do not contain high levels of the sulfur-containing amino acids, cysteine and methionine. Ultimately, unraveling the fundamental steps and regulation of thiol metabolism in soybean is important for optimizing crop yield and quality. Here we review the pathways from sulfur uptake to glutathione and homoglutathione synthesis in soybean, the potential biotechnology benefits of understanding and modifying these pathways, and how information from the soybean genome may guide the next steps in exploring this biochemical system.


Genome-wide DNA methylation of the liver reveals delayed effects of early-life exposure to 17-α-ethinylestradiol in the self-fertilizing mangrove rivulus.
Organisms exposed to endocrine disruptors in early life can show altered phenotype later in adulthood. Although the mechanisms underlying these long-term effects remain poorly understood, an increasing body of evidence points towards the potential role of epigenetic processes. In the present study, we exposed hatchlings of an isogenic lineage of the self-fertilizing fish mangrove rivulus for 28 days to 4 and 120 ng/L of 17-α-ethinylestradiol. After a recovery period of 140 days, reduced representation bisulphite sequencing (RRBS) was performed on the liver in order to assess the hepatic genome-wide methylation landscape. Across all treatment comparisons, a total of 146 differentially methylated fragments (DMFs) were reported, mostly for the group exposed to 4 ng/L, suggesting a non-monotonic effect of EE2 exposure. Gene ontology analysis revealed networks involved in lipid metabolism, cellular processes, connective tissue function, molecular transport and inflammation. The highest effect was reported for nipped-B-like protein B (NIPBL) promoter region after exposure to 4 ng/L EE2 (+ 21.9%), suggesting that NIPBL could be an important regulator for long-term effects of EE2. Our results also suggest a significant role of DNA methylation in intergenic regions and potentially in transposable elements. These results support the ability of early exposure to endocrine disruptors of inducing epigenetic alterations during adulthood, providing plausible mechanistic explanations for long-term phenotypic alteration. Additionally, this work demonstrates the usefulness of isogenic lineages of the self-fertilizing mangrove rivulus to better understand the biological significance of long-term alterations of DNA methylation by diminishing the confounding factor of genetic variability.


Health-related quality of life with hereditary angioedema following prophylaxis with subcutaneous C1-inhibitor with recombinant hyaluronidase.
To estimate health-related quality-of-life changes in patients with hereditary angioedema due to C1-inhibitor (C1-INH) deficiency who received subcutaneous C1-INH with recombinant hyaluronidase (rHuPH20) for attack prophylaxis in a randomized, double-blind, dose-ranging, cross-over study. Patients with type I/II hereditary angioedema received 1000 U of C1-INH with 24,000 U of rHuPH20 or 2000 U of C1-INH with 48,000 U of rHuPH20 every 3-4 days for 8 weeks and then crossed over for another 8-week period. The study was terminated early as a precaution related to non-neutralizing antibodies to rHuPH20. The Angioedema Quality of Life questionnaire (AE-QoL) was administered at weeks 1 and 5 of both periods, and at 1 week after the second treatment period. Changes in AE-QoL scores were calculated over both treatment periods and within each treatment period for patients with ≥4 weeks of treatment. Forty-one patients had evaluable AE-QoL data, and 22 patients completed treatment. At screening, 43% of the patients were receiving intravenous C1-INH. A significant average AE-QoL total score decline (improvement) of -8.1 (95% confidence interval, -13.7 to -2.5) was observed from baseline to the end of the study, and significant AE-QoL score declines were observed in the Functioning, Fear/Shame, and Nutrition domains. Patients on 2000 U reported higher mean AE-QoL score declines in Functioning and Nutrition domains relative to the 1000 U dose. Overall, 43.9% of all the patients, 45.5% of the study completers, and 46.7% of the nonprophylaxis users at baseline on high treatment doses achieved a reduction in the AE-QoL total score of six points. Despite early termination and prestudy prophylactic intravenous C1-INH use by 43% of the patients, improved AE-QoL scores were observed after ≤16 weeks of subcutaneous C1-INH-rHuPH20 prophylaxis.


Enhanced flux of substrates into polyamine biosynthesis but not ethylene in tomato fruit engineered with yeast S-adenosylmethionine decarboxylase gene.
S-adenosylmethionine (SAM), a major substrate in 1-C metabolism is a common precursor in the biosynthetic pathways of polyamines and ethylene, two important plant growth regulators, which exhibit opposing developmental effects, especially during fruit ripening. However, the flux of various substrates including SAM into the two competing pathways in plants has not yet been characterized. We used radiolabeled (14)C-Arg, (14)C-Orn, L-[U-(14)C]Met, (14)C-SAM and (14)C-Put to quantify flux through these pathways in tomato fruit and evaluate the effects of perturbing these pathways via transgenic expression of a yeast SAM decarboxylase (ySAMDC) gene using the fruit ripening-specific promoter E8. We show that polyamines in tomato fruit are synthesized both from Arg and Orn; however, the relative contribution of Orn pathway declines in the later stages of ripening. Expression of ySAMDC reversed the ripening associated decline in spermidine (Spd) and spermine (Spm) levels observed in the azygous control fruit. About 2- to 3-fold higher levels of labeled-Spd in transgenic fruit (556HO and 579HO lines) expressing ySAMDC confirmed the enzymatic function of the introduced gene. The incorporation of L-[U-(14)C]Met into Spd, Spm, ethylene and 1-aminocyclopropane-1-carboxylic acid (ACC) was used to determine Met-flux into these metabolites. The incorporation of (14)C-Met into Spd/Spm declined during ripening of the control azygous fruit but this was reversed in fruits expressing ySAMDC. However, incorporation of (14)C-Met into ethylene or ACC during ripening was not altered by the expression of ySAMDC in the fruit. Taken together these results show that: (1) There is an inverse relationship between the production of higher polyamines and ethylene during fruit ripening, (2) the inverse relationship between higher polyamines and ethylene is modulated by ySAMDC expression in that the decline in Spd/Spm during fruit ripening can be reversed without significantly altering ethylene biosynthesis, and (3) cellular flux of SAM in plants is homeostatically regulated based on its demand for competing pathways.


Is it my job? Alcohol brief interventions: knowledge and attitudes among future health-care professionals in Scotland.
To document knowledge and perceptions relating to the professional role in the area of alcohol misuse within a sample of first year (n = 278) and final year (n = 527) medical, nursing and allied health professional (NAHP) students in Scotland. A cross sectional survey design involving self-completed questionnaires administered in autumn 2008 (first year students) and spring 2009 (final year students) through course websites and lectures. Gaps in the knowledge relating to current UK health guidelines were identified but more so among NAHP students than medical students. Exploration of the perceived role in this area of practice identified three broad groups of students: those clear about their role (medical and nursing students), those advocating a role but not identified by fellow students (occupational therapy and pharmacy) and those uncertain of their role (radiographers, speech and language therapists/audiologists and physiotherapy). Higher education institutions should address the gaps in the knowledge around guidance for alcohol consumption. The effectiveness of brief interventions may depend on it. Additionally, through inter-professional teaching and in collaboration with the relevant professional bodies, more could be done to promote the contribution of practitioners other than those traditionally linked (i.e. medical and nursing) to this important clinical role.


Functional expression of the alpha 7 integrin receptor in differentiated smooth muscle cells.
Expression of the alpha7 integrin is developmentally regulated and is thought to be tissue-specific for both skeletal and cardiac muscles. We now report that alpha7 is also strongly and ubiquitously expressed by various types of smooth muscle, including vascular, gastrointestinal and genitourinary smooth muscles. In addition, alpha7 was surface-expressed by a number of smooth muscle cell lines that maintained their differentiated phenotype following adaptation to culture. Studies with the mouse 9E11G smooth muscle cell line showed that the alpha7 integrin mediated both adhesion and motility of these cells on laminin 1 substrates. Alpha7 expression appears to correlate with the smooth-muscle-differentiated phenotype. The multipotential P19 mouse embryonic stem cell line lacks alpha7 but uses the alpha6 integrin to adhere to laminin 1. Following retinoic acid-induced P19 differentiation predominantly to the smooth muscle cell lineage, high expression of alpha7 was detected along with partial dependence on alpha7 for binding to laminin. The expression of alpha7 paralleled the induction of smooth-muscle-specific alpha-actin, as revealed by dual-labeling flow cytometry. In contrast, alpha7, which initially was highly expressed on the surface of vascular smooth muscle cell explants, was rapidly downregulated in smooth muscle cell outgrowths as they dedifferentiated into their synthetic phenotype. The results indicate that the expression of alpha7 integrin in smooth muscle cells is associated with their differentiated phenotype and mediates their interaction with laminins.


Expression of a novel gene, MafB, in Dupuytren's disease.
Dupuytren's disease (DD) is characterized by fibroblastic proliferation of the palmar fascia, often leading to flexion contracture in the hand. Although there is a strong genetic component the genome-wide expression of novel genes is not known. The purpose of this study was to use DNA microarray technology to identify upregulated genes in DD. Human tissue samples were harvested from 3 patient sources: DD cord tissue (n = 20), normal-appearing adjacent control fascia (n = 15), and palmar fascia from patients having carpal tunnel release (n = 15). DNA microarray analysis was performed on amplified sample RNA. Novel genes were compared with known gene functions. A candidate gene of interest was studied further by using immunohistochemistry on DD tissue samples and controls. Several novel genes not described previously in the study of DD were upregulated significantly, including MafB, collagen type V, alpha-2 (COL5A2), collagen type VIII, alpha-1 (COL8A1), contactin I (CNTN1), and leucine-rich repeat containing 17 (LRRC17). These upregulated genes were compared with their known gene-expression profiles in other tissues and their purported functions. MafB was found to be of particular interest because of its prominent role in tissue development and cellular differentiation. MafB immunohistochemistry showed positive staining in 50% of the DD specimens but complete absence of MafB in all control tissues (adjacent control fascia, carpal tunnel fascia). Co-localization experiments with MafB and alpha-smooth muscle actin showed staining properties in similar regions but these 2 proteins were not confined solely to the same cells. Microarray analysis of DD tissue has identified significant upregulated gene expression of MafB. MafB protein also is found in Dupuytren's cords but not in control fascia. Co-localization data suggest that the association of MafB with DD is not related exclusively to myofibroblast proliferation. Because of its role in fibroblastic transformation in other models MafB and its relationship to the pathogenesis of DD deserves further study.


Biophysical characterisation of fibulin-5 proteins associated with disease.
FBLN5 encodes fibulin-5, an extracellular matrix calcium-binding glycoprotein that is essential for elastic fibre formation. FBLN5 mutations are associated with two distinct human diseases, age-related macular degeneration (AMD) and cutis laxa (CL), but the biochemical basis for the pathogenic effects of these mutations is poorly understood. Two missense mutations found in AMD patients (I169T and G267S) and two missense mutations found in CL patients (G202R and S227P) were analysed in a native-like context in recombinant fibulin-5 fragments. Limited proteolysis, NMR spectroscopy and chromophoric calcium chelation experiments showed that the G267S and S227P substitutions cause long-range structural effects consistent with protein misfolding. Cellular studies using fibroblast cells further demonstrated that these recombinant forms of mutant fibulin-5 were not present in the extracellular medium, consistent with retention. In contrast, no significant effects of I169T and G202R substitutions on protein fold and secretion were identified. These data establish protein misfolding as a causative basis for the effects of G267S and S227P substitutions in AMD and CL, respectively, and raise the possibility that the I169T and G202R substitutions may be polymorphisms or may increase susceptibility to disease.


Search for polyoma-, herpes-, and bornaviruses in squirrels of the family Sciuridae.
Squirrels (family Sciuridae) are globally distributed members of the order Rodentia with wildlife occurrence in indigenous and non-indigenous regions (as invasive species) and frequent presence in zoological gardens and other holdings. Multiple species introductions, strong inter-species competition as well as the recent discovery of a novel zoonotic bornavirus resulted in increased research interest on squirrel pathogens. Therefore we aimed to test a variety of squirrel species for representatives of three virus families. Several species of the squirrel subfamilies Sciurinae, Callosciurinae and Xerinae were tested for the presence of polyomaviruses (PyVs; family Polyomaviridae) and herpesviruses (HVs; family Herpesviridae), using generic nested polymerase chain reaction (PCR) with specificity for the PyV VP1 gene and the HV DNA polymerase (DPOL) gene, respectively. Selected animals were tested for the presence of bornaviruses (family Bornaviridae), using both a broad-range orthobornavirus- and a variegated squirrel bornavirus 1 (VSBV-1)-specific reverse transcription-quantitative PCR (RT-qPCR). In addition to previously detected bornavirus RNA-positive squirrels no more animals tested positive in this study, but four novel PyVs, four novel betaherpesviruses (BHVs) and six novel gammaherpesviruses (GHVs) were identified. For three PyVs, complete genomes could be amplified with long-distance PCR (LD-PCR). Splice sites of the PyV genomes were predicted in silico for large T antigen, small T antigen, and VP2 coding sequences, and experimentally confirmed in Vero and NIH/3T3 cells. Attempts to extend the HV DPOL sequences in upstream direction resulted in contiguous sequences of around 3.3 kilobase pairs for one BHV and two GHVs. Phylogenetic analysis allocated the novel squirrel PyVs to the genera Alpha- and Betapolyomavirus, the BHVs to the genus Muromegalovirus, and the GHVs to the genera Rhadinovirus and Macavirus. This is the first report on molecular identification and sequence characterization of PyVs and HVs and the detection of bornavirus coinfections with PyVs or HVs in two squirrel species. Multiple detection of PyVs and HVs in certain squirrel species exclusively indicate their potential host association to a single squirrel species. The novel PyVs and HVs might serve for a better understanding of virus evolution in invading host species in the future.


A pilot study using magnetic resonance imaging to determine the pattern of muscle group recruitment by rowers with different levels of experience.
To determine whether it was possible using magnetic resonance imaging (MRI) to define the pattern of muscle recruitment in a specific sport (rowing) and to see whether there were differences in this pattern between athletes of different experience. It has been shown that during vigorous exercise the water content of muscle increases transiently. This can be observed using MRI, where the prolonged T2 relaxation time of muscle can be demonstrated. In this study we have exploited the increase in signal seen in exercised muscle on short TI inversion recovery (STIR) sequences, to show how rowers of different experience use different muscle groups. We have shown that trained athletes recruit selected muscle groups to carry out a given task, which they carry out more efficiently than untrained or less experienced athletes. We have provided the basis of potential research to refine training methods, in order to develop specific muscle groups in athletes, in the hope of achieving a higher level of performance at an earlier stage in their training. We have also defined a technique that may be of clinical value in cases of muscle dysfunction.


Antibacterial activity of propolis extracts from the south of Portugal.
To examine the antibacterial activity of diverse extracts of propolis harvested at winter and spring from several locations of Algarve, Portugal, against Gram-negative and Gram-positive bacteria was the main goal of the present work. For such, the antibacterial activity was determined by agar diffusion. The results showed that all tested bacterial strains showed susceptibility to diluted propolis extracts and in a dose-dependent manner. Two propolis samples collected at springtime showed higher antibacterial activity, in comparison with samples harvested at wintertime. Ethanolic and methanolic extracts have a very similar activity (P<0.05). Helicobacter pylori strains J99 and 26695 were the most susceptible strains to the tested extracts (33.67±2.52 mm and 35.67±0.58mm, respectively). This study constitutes the first approach of the biological activities of Portuguese propolis from the Algarve region and evidences its potential use to combat bacterial infections, in particular against the gastric pathogen H. pylori.


Toxicity associated with adjuvant postoperative therapy for adenocarcinoma of the rectum.
The Gastrointestinal Tumor Study Group's (GITSG) adjuvant rectal carcinoma study compared four postoperative treatment regimens: (1) control (no adjuvant therapy); (2) chemotherapy alone consisting of pulses of 5-fluorouracil and methyl CCNU for 18 months; (3) pelvic and perineal radiotherapy using parallel opposed fields with 4000 rad in 4.5 to 5 weeks or 4800 rad in 5 to 5.5 weeks; and (4) a combination of both modalities. The results of this study are published elsewhere and show a significantly reduced recurrence rate and prolonged disease-free survival time for the combined modality arm compared with the no therapy arm. Severe toxicity in the combined therapy arm was significantly worse (P less than 0.001) than in either single modality arm. Most of the differences in toxicity experienced between the three regimens involved diarrhea, thrombocytopenia, and leukopenia. Analysis of all parameters of radiotherapy quality assurance data was not significantly associated with toxicity. Radiation enteritis was noted in 5 patients of 96 (5.2%) in the two arms containing irradiation. All five required laparotomy. The two enteritis fatalities occurred late at 605 and 1000 days after start of combined modality treatment, respectively. One other patient on the chemotherapy arm died of acute nonlymphocytic leukemia. The authors conclude that combined radiotherapy and chemotherapy, although significantly more effective in reducing recurrence than no therapy, is significantly more toxic than single-modality therapy in many parameters, although most of the toxicity is transient and therefore not limiting. Late complications, which are less reversible and therefore much more important than early reactions, and radiation enteritis in this study were relatively uncommon. This schedule of combined modality therapy is not only effective but appears to have tolerable toxicity, because of the relative lack of late effects.


Neuronal population oscillations of rat hippocampus during epileptic seizures.
Neuronal population oscillations in the hippocampus have an important effect in the information processing in the brain and the generation of epileptic seizures. In this paper, we investigate the neuronal population oscillations in the hippocampus of epileptic rats in vivo using an empirical mode decomposition (EMD) method. A neuronal population oscillation can be decomposed into several relaxation oscillations, which possess a recovery and release phase, with the different frequencies that ranges from 0 to 600 Hz. The natures of relaxation oscillations at the pre-ictal, seizure onset and ictal states are distinctly different. The analysis of relaxation oscillations show that the gamma wave is a lead relaxation oscillation at the pre-ictal stage, then it moves to beta oscillation or theta oscillation while the ictal stage starts; the fast relaxation oscillations are associated with the slow relaxation oscillations in the CA1 or CA3, in particular, the fast relaxation oscillations are associated on the recovery phase of the slow relaxation oscillations during the pre-ictal interval, however move to the release phase of the slow relaxation oscillations during the ictal interval. Comparison of the relaxation oscillations in CA1 and CA3 shows that the neurons in the CA1 are more active during the epileptic seizures than during the pre-ictal stage. These findings demonstrate that this method is very helpful to decompose neuronal population for understanding the underlying mechanism of epileptic seizures.


Generation of HIV-1-specific T cells by electroporation of T-cell receptor RNA.
HIV-1-specific cytotoxic T lymphocytes, which recognize conserved epitopes of the virus, are correlated with prolonged survival of infected individuals. Unfortunately, most HIV-1-infected patients are unable to generate such an immune response. Antigen-specific cytotoxic T lymphocytes can be generated by T-cell receptor transfer. This is commonly done by retroviral transduction, which is complicated and poses the threat of stable genetic alteration of autologous cells. We reprogrammed primary CD8+ T cells by electroporation of RNA, which encoded an HIV-1-pol- and an HIV-1-gag-specific T-cell receptor recognizing the human leukocyte antigen-A2 restricted epitopes ILKEPVHGV and SLYNTVATL, respectively. These reprogrammed cells specifically produced the proinflammatory cytokines interleukin-2, tumor necrosis factor-alpha, and interferon-gamma after stimulation with target cells that presented the corresponding peptides, and were able to lyse these targets efficiently and specifically. The lytic avidities of the HIV-1-pol- and HIV-1-gag-TCR-RNA-electroporated CD8+ T cells were within the same range than those of the parental cytotoxic T lymphocytes. Most importantly, HIV-1-gag-reprogrammed T cells recognized target cells that presented endogenously processed antigen, which resulted in cytokine production and lysis. It is shown here for the first time that functional transfer of virus-specific T-cell receptors by RNA electroporation is feasible, and represents an innovative, safe, and easy method to generate virus-specific T cells, avoiding the risks of retroviral transduction.