Revisiting the extracellular lifestyle.
In microbiology textbooks infectious agents are traditionally classified as intracellular or extracellular pathogens depending on whether they multiply inside or outside host cells. In recent literature an increasing number of extracellular pathogens are described in close apposition with the host cell surface embedded in plasma membrane folds, making it difficult to classify them as strictly extracellular pathogens. This review further explores this emerging new lifestyle category tentatively named 'epicellular' in reference to earlier work describing the location of the parasite Cryptosporidium parvum. The lifestyles of three diverse such pathogens were examined: the parasite Cryptosporidium parvum, the Gram-negative bacterium Neisseria meningitidis and the human T cell leukaemia virus type 1 (HTLV-1). The specific cellular location, the mechanisms of adhesion, the induction of plasma membrane folds and the subsequent functional consequences will be compared. Although current knowledge suggests different underlying mechanisms, a concept that emerges is that the particular location of these pathogens has important functional consequences for the pathogens in terms of nutrient acquisition, immune escape, resistance to mechanical stress and transmission. Re-examining the lifestyle of other classically extracellular pathogens might thus shed a new light on the way pathogens interact with cells and point to new areas of investigation.


Prevalence of HIV and Treponema pallidum infections in pregnant women in Campinas and their association with socio-demographic factors.
The anonymous seroprevalence of HIV and syphilis was studied by collecting umbilical cord blood samples from 5,815 women who gave birth in Campinas' hospitals throughout a six-month period. ELISA and Western blot were used for HIV, and VDRL and TPHA for Treponema pallidum screening. While maintaining the anonymity of the women, information was recorded on the hospital of origin, divided into university (public) and private hospitals, as well as on the form of payment (social security, private insurance or direct payment), age, marital status, education, employment and place of residence. Seroprevalence was 0.42 percent for HIV and 1.16 percent for syphilis. There was a significant correlation between a positive reaction to the two infections (p = 0.02). After univariate and logistic regression analysis, only university hospitals were shown to be associated with seropositivity for HIV, whereas the same variable and an older age were associated with syphilis. All positive reactions were found either in public hospitals or among social security patients treated at private institutions. The conclusion was that HIV infection is becoming almost as prevalent as syphilis among this population, and affects primarily the lower socio-economic strata. This suggests that routine, voluntary HIV serology should be considered and discussed with patients during prenatal or delivery care whenever a population shows a seroprevalence close to or greater than 1 percent.


Effect of stage-based education provided by dedicated dietitians on hyperphosphataemic haemodialysis patients: results from the Nutrition Education for Management of Osteodystrophy randomised controlled trial.
The Nutrition Education for Management of Osteodystrophy trial showed that stage-based nutrition education by dedicated dietitians surpasses existing practices in Lebanon with respect to lowering serum phosphorus among general haemodialysis patients. The present study explores the effect of nutrition education specifically on hyperphosphataemic patients from this trial. Hyperphosphataemic haemodialysis patients were allocated to a dedicated dietitian (DD), a trained hospital dietitian (THD) and existing practice (EP) protocols. From time-point (t)-0 until t-1 (6 months), the DD group (n = 47) received 15 min of biweekly nutrition education by dedicated dietitians trained on renal nutrition; the THD group (n = 89) received the usual care from trained hospital dietitians; and the EP group (n = 42) received the usual care from untrained hospital dietitians. Patients were followed-up from t-1 until t-2 (6 months). Analyses used two-way repeated measures analysis of variance and Cohen's effect sizes (d). At t-1, phosphataemia significantly decreased in all groups (DD:-0.27 mmol L-1 ; EP:-0.15 mmol L-1 ; THD:-0.12 mmol L-1 ; P < 0.05); the DD protocol had the greatest effect relative to EP (d = -0.35) and THD (d = -0.50). Only the DD group showed more readiness to adhere to a low phosphorus diet at t-1; although, at t-2, this regressed to baseline levels. The malnutrition inflammation score remained stable only in the DD group, whereas the EP and THD groups exhibited a significant increase (DD: 6.74, 6.97 and 7.91; EP: 5.82, 8.69 and 8.13; THD: 5.33, 7.92 and 9.42, at t-0, t-1 and t-2, respectively). The results of the present study suggest that the DD protocol decreases serum phosphorus compared to EP and THD, at the same time as maintaining the nutritional status of hyperphosphataemic haemodialysis patients. Assessing the cost-effectiveness of the DD protocol is recommended.


Properties of the exact universal functional in multicomponent density functional theory.
Multicomponent density functional theory has been developed to treat systems with more than one type of quantum particle, such as electrons and nuclei, in an external potential. The existence of the exact universal multicomponent density functional in terms of the one-particle densities for each type of quantum particle has been proven. In the present paper, a number of important mathematical properties of the exact universal multicomponent density functional are derived. The expression relating the electron-proton pair density to the one-particle densities leads to an inequality for the potential energy component of the electron-proton correlation functional under well-defined conditions. General inequalities for the kinetic energy correlation functionals and the total electron-proton correlation functional are also derived. The coordinate scaling analysis leads to mathematical inequalities describing the effect of scaled densities on the kinetic, potential, and total energy functionals. The adiabatic connection formula defines the exact electron-proton functional in terms of an adiabatic scaling parameter that smoothly connects the noninteracting system with the fully interacting system. The virial expression provides the relation between the exact kinetic and potential energy functionals for the ground state densities of multicomponent systems. These mathematical relationships provide insight into the fundamental properties of the exact universal multicomponent density functional and serve as a guide for the development of approximate electron-proton density functionals.


Small Sample Kernel Association Tests for Human Genetic and Microbiome Association Studies.
Kernel machine based association tests (KAT) have been increasingly used in testing the association between an outcome and a set of biological measurements due to its power to combine multiple weak signals of complex relationship with the outcome through the specification of a relevant kernel. Human genetic and microbiome association studies are two important applications of KAT. However, the classic KAT framework relies on large sample theory, and conservativeness has been observed for small sample studies, especially for microbiome association studies. The common approach for addressing the small sample problem relies on computationally intensive resampling methods. Here, we derive an exact test for KAT with continuous traits, which resolve the small sample conservatism of KAT without the need for resampling. The exact test has significantly improved power to detect association for microbiome studies. For binary traits, we propose a similar approximate test, and we show that the approximate test is very powerful for a wide range of kernels including common variant- and microbiome-based kernels, and the approximate test controls the type I error well for these kernels. In contrast, the sequence kernel association tests have slightly inflated genomic inflation factors after small sample adjustment. Extensive simulations and application to a real microbiome association study are used to demonstrate the utility of our method.


Transcranial Direct Current Stimulation to Facilitate Lower Limb Recovery Following Stroke: Current Evidence and Future Directions.
Stroke remains a global leading cause of disability. Novel treatment approaches are required to alleviate impairment and promote greater functional recovery. One potential candidate is transcranial direct current stimulation (tDCS), which is thought to non-invasively promote neuroplasticity within the human cortex by transiently altering the resting membrane potential of cortical neurons. To date, much work involving tDCS has focused on upper limb recovery following stroke. However, lower limb rehabilitation is important for regaining mobility, balance, and independence and could equally benefit from tDCS. The purpose of this review is to discuss tDCS as a technique to modulate brain activity and promote recovery of lower limb function following stroke. Preliminary evidence from both healthy adults and stroke survivors indicates that tDCS is a promising intervention to support recovery of lower limb function. Studies provide some indication of both behavioral and physiological changes in brain activity following tDCS. However, much work still remains to be performed to demonstrate the clinical potential of this neuromodulatory intervention. Future studies should consider treatment targets based on individual lesion characteristics, stage of recovery (acute vs. chronic), and residual white matter integrity while accounting for known determinants and biomarkers of tDCS response.


Circadian programs in cyanobacteria: adaptiveness and mechanism.
At least one group of prokaryotes is known to have circadian regulation of cellular activities--the cyanobacteria. Their "biological clock" orchestrates cellular events to occur in an optimal temporal program, and it can keep track of circadian time even when the cells are dividing more rapidly than once per day. Growth competition experiments demonstrate that the fitness of cyanobacteria is enhanced when the circadian period matches the period of the environmental cycle. Three genes have been identified that specifically affect circadian phenotypes. These genes, kaiA, kaiB, and kaiC, are adjacent to each other on the chromosome, thus forming a clock gene cluster. The clock gene products appear to interact with each other and form an autoregulatory feedback loop.


Stakeholders' opinions on a future in-vehicle alcohol detection system for prevention of drunk driving.
There is a common understanding that driving under the influence of alcohol is associated with higher risk of being involved in crashes with injuries and possible fatalities as the outcome. Various countermeasures have therefore from time to time been taken by the authorities to prevent drunk driving. One of them has been the alcohol interlock. Up to now, interlocks have mainly been used by previously convicted drunk drivers and in the commercial road transport sector, but not in private cars. New technology has today reached a level where broader implementation might be possible. To our knowledge, however, little is known about different stakeholders' opinions of a broader implementation of such systems. In order to increase that knowledge, we conducted a focus group study to collect in-depth thoughts from different stakeholders on this topic. Eight focus groups representing a broad societal span were recruited and conducted for the purpose. The results show that most stakeholders thought that an integrated system for alcohol detection in vehicles might be beneficial in lowering the number of drunk driving crashes. They said that the system would probably mainly prevent driving by people who unintentionally and unknowingly drive under the influence of alcohol. The groups did, however, not regard the system as a final solution to the drunk driving problem, and believed that certain groups, such as criminals and alcoholics, would most likely find a way around the system. Concerns were raised about the risk of increased sleepy driving and driving just under the legal blood alcohol concentration (BAC) limit. The results also indicate that stakeholders preferred a system that provides information on the BAC up to the legal limit, but not for levels above the limit; for those, the system should simply prevent the car from starting. Acceptance of the system depended on the reliability of the system, on its ability to perform fast sampling, and on the analytical process, as well as the system's more or less inconspicuous placement and user-friendliness. The stakeholders thought that drivers would probably not voluntarily demand the system. So if broad implementation was desired, it would have to be made compulsory by legislation. As an incentive to increase demand, lower taxes and insurance premiums were suggested.


Are monkeys sensitive to informativeness: An experimental study with baboons (Papio papio).
Informativeness (defined as reduction of uncertainty) is central in human communication. In the present study, we investigate baboons' sensitivity to informativeness by manipulating the informativity of a cue relative to a response display and by allowing participants to anticipate their answers or to wait for a revealed answer (with variable delays). Our hypotheses were that anticipations would increase with informativity, while response times to revealed trials would decrease with informativity. These predictions were verified in Experiment 1. In Experiments 2 and 3, we manipulated rewards (rewarding anticipation responses at 70% only) to see whether reward tracking alone could account for the results in Experiment 1. We observed that the link between anticipations and informativeness disappeared, but not the link between informativeness and decreased RTs for revealed trials. Additionally, in all three experiments, the number of correct answers in revealed trials with fast reaction times (< 250ms) increased with informativeness. We conclude that baboons are sensitive to informativeness as an ecologically sound means to tracking reward.


A musculoskeletal shoulder model based on pseudo-inverse and null-space optimization.
The goal of the present work was assess the feasibility of using a pseudo-inverse and null-space optimization approach in the modeling of the shoulder biomechanics. The method was applied to a simplified musculoskeletal shoulder model. The mechanical system consisted in the arm, and the external forces were the arm weight, 6 scapulo-humeral muscles and the reaction at the glenohumeral joint, which was considered as a spherical joint. The muscle wrapping was considered around the humeral head assumed spherical. The dynamical equations were solved in a Lagrangian approach. The mathematical redundancy of the mechanical system was solved in two steps: a pseudo-inverse optimization to minimize the square of the muscle stress and a null-space optimization to restrict the muscle force to physiological limits. Several movements were simulated. The mathematical and numerical aspects of the constrained redundancy problem were efficiently solved by the proposed method. The prediction of muscle moment arms was consistent with cadaveric measurements and the joint reaction force was consistent with in vivo measurements. This preliminary work demonstrated that the developed algorithm has a great potential for more complex musculoskeletal modeling of the shoulder joint. In particular it could be further applied to a non-spherical joint model, allowing for the natural translation of the humeral head in the glenoid fossa.


Disease-modifying drugs can reduce disability progression in relapsing multiple sclerosis.
An ever-expanding number of disease-modifying drugs for multiple sclerosis have become available in recent years, after demonstrating efficacy in clinical trials. In the real-world setting, however, disease-modifying drugs are prescribed in patient populations that differ from those included in pivotal studies, where extreme age patients are usually excluded or under-represented. In this multicentre, observational, retrospective Italian cohort study, we evaluated treatment exposure in three cohorts of patients with relapsing-remitting multiple sclerosis defined by age at onset: paediatric-onset (≤18 years), adult-onset (18-49 years) and late-onset multiple sclerosis (≥50 years). We included patients with a relapsing-remitting phenotype, ≥5 years follow-up, ≥3 Expanded Disability Status Scale (EDSS) evaluations and a first neurological evaluation within 3 years from the first demyelinating event. Multivariate Cox regression models (adjusted hazard ratio with 95% confidence intervals) were used to assess the risk of reaching a first 12-month confirmed disability worsening and the risk of reaching a sustained EDSS of 4.0. The effect of disease-modifying drugs was assessed as quartiles of time exposure. We found that disease-modifying drugs reduced the risk of 12-month confirmed disability worsening, with a progressive risk reduction in different quartiles of exposure in paediatric-onset and adult-onset patients [adjusted hazard ratios in non-exposed versus exposed >62% of the follow-up time: 8.0 (3.5-17.9) for paediatric-onset and 6.3 (4.9-8.0) for adult-onset, P < 0.0001] showing a trend in late-onset patients [adjusted hazard ratio = 1.9 (0.9-4.1), P = 0.07]. These results were confirmed for a sustained EDSS score of 4.0. We also found that relapses were a risk factor for 12-month confirmed disability worsening in all three cohorts, and female sex exerted a protective role in the late-onset cohort. This study provides evidence that sustained exposure to disease-modifying drugs decreases the risk of disability accumulation, seemingly in a dose-dependent manner. It confirms that the effectiveness of disease-modifying drugs is lower in late-onset patients, although still detectable.


Primary valve malfunction of the Krupin eye valve with disk.
To examine the management and possible causes of primary valve malfunction of the Krupin eye valve with disk. The authors reviewed the results of 113 patients undergoing implantation of the Krupin eye valve with disk and identified eight patients with primary valve malfunction requiring surgical revision. Valve revision involved manipulation (n = 1 case), explantation of the malfunctioning valve and implantation of a new valve (n = 2), and amputation of the valve (n = 5). Six of eight patients had final intraocular pressures of < 21 mmHg on one or no medications at a mean interval of 15.9 months (range 5-36) after surgical revision. Transient postoperative hypotony was noted in three patients and chronic hypotony with loss of light perception in one patient. One explanted valve was examined and found to have partially fused leaflets. Surgical revision in cases of primary valve malfunction of the Krupin eye valve with disk may be accomplished relatively safely with an acceptable level of postoperative complications. The etiology of primary valve malfunction may be related to the sterilization process and prolonged storage before implantation.


Deciphering nifedipine in vivo delivery from modified release dosage forms: Identification of food effect.
With the increased reliance on in vitro dissolution testing as an indicator of in vivo drug behavior and the trend towards the in silico modeling of dosage form performance, the need for bioperformance dissolution methodology development has been enhanced. Determination of the in vivo drug delivery profile is essential for the bioperformance dissolution test development and in vitro/in vivo correlation modeling, as well as the understanding of absorption mechanisms. The aim of this study was to compare different methods in terms of their usefulness and applicability in deciphering in vivo delivery of nifedipine administered in modified release dosage forms. A detailed survey of publications on nifedipine pharmacokinetics was done and used to identify the magnitude of food effect. In vitro dissolution testing was performed under various experimental conditions. Obtained results indicate the potential for using the developed in silico model coupled with discriminative in vitro dissolution data for identification of the in vivo drug product behavior.


Geographic variation in host-specificity and parasitoid pressure of an herbivore (geometridae) associated with the tropical genus piper (piperaceae).
The extraordinary diversity of tropical herbivores may be linked to hostplant specialization driven in part by variation in pressure from natural enemies. We quantified levels of host-specificity and parasitoid attack for the specialist herbivore, Eois (Geometridae). The goals of this research were to examine: 1) whether Eois are specialized on the genus Piper (Piperaceae) and if hostplant specialization varies geographically; 2) whether Eois are equally vulnerable to parasitoid attack across different geographic regions and by the same parasitoid families; and 3) whether parasitism levels vary with precipitation and elevation. Based on over 15,000 rearings, we found Eois caterpillars feeding exclusively on Piper. However, we did not detect geographic differences in host-specificity; each Eois species fed on an average of two Piper species. Parasitism levels of Eois varied significantly with climate and topography; Eois were most vulnerable to parasitoid attack in moist versus dry and wet forests and at low versus high elevations. The diversity of parasitoid families reared from Eois was greater in Ecuador and Costa Rica than in Panama, where parasitoids were primarily in the family Braconidae. The quantitative evidence for host-specificity provides support for the hypothesis that Eois are specialized on Piper. Our results also reveal that Eois are exposed to a mosaic of potential selective pressures due to variation in parasitoid attack over a large spatial scale.


Putting your course on the Web: lessons from a case study and systematic literature review.
Education via the Internet offers enormous potential, but many online courses are pedagogically or technically weak and many good projects are never mainstreamed. In drawing up our recommendations to address the issues around putting a course on the web, we drew on 3 main sources of data: an extensive in-depth course evaluation; a systematic review of the literature, and questions raised by participants on our training-the-trainers courses. For any web-based course to succeed, 10 overlapping and iterative areas of activity must be addressed. These are: the market for the course; course aims and intended learning outcomes; choice of software platform; staff training needs; writing high quality study materials; design features for active learning; technical and administrative challenges; evaluation and quality improvement; mainstreaming the course within the institution, and financial viability.


Accidental nuclear war--a post-cold war assessment.
In the 1980s, many medical organizations identified the prevention of nuclear war as one of the medical profession's most important goals. An assessment of the current danger is warranted given the radically changed context of the post-Cold War era. We reviewed the recent literature on the status of nuclear arsenals and the risk of nuclear war. We then estimated the likely medical effects of a scenario identified by leading experts as posing a serious danger: an accidental launch of nuclear weapons. We assessed possible measures to reduce the risk of such an event. U.S. and Russian nuclear-weapons systems remain on a high-level alert status. This fact, combined with the aging of Russian technical systems, has recently increased the risk of an accidental nuclear attack. As a conservative estimate, an accidental intermediate-sized launch of weapons from a single Russian submarine would result in the deaths of 6,838,000 persons from firestorms in eight U.S. cities. Millions of other people would probably be exposed to potentially lethal radiation from fallout. An agreement to remove all nuclear missiles from high-level alert status and eliminate the capability of a rapid launch would put an end to this threat. The risk of an accidental nuclear attack has increased in recent years, threatening a public health disaster of unprecedented scale. Physicians and medical organizations should work actively to help build support for the policy changes that would prevent such a disaster.


Systemic lupus erythematosus in three ethnic groups. IV. Factors associated with self-reported functional outcome in a large cohort study. LUMINA Study Group. Lupus in Minority Populations, Nature versus Nurture.
To identify features of systemic lupus erythematosus (SLE) associated with poor functional outcome as measured by the 36-item Medical Outcomes Study Short Form 36 Health Survey (SF-36). Two hundred twenty-four patients with early SLE (70 Hispanic, 83 African American, and 71 white) enrolled in a longitudinal study of outcomes were evaluated at study entry. The 8 composite scales and 2 summary measures (physical and mental) of the SF-36 were the dependent variables. Independent variables--1) sociodemographic, 2) clinical features, 3) immunologic, 4) global scores, and 5) behavioral/cultural--were examined for each of the scales and summary measures and for each ethnic group. Significant variables in these analyses were then used to construct models to determine their association with each of the scales and the 2 summary measures for the entire population and the 3 ethnic groups. Self-reported physical and mental functioning were most consistently associated with abnormal illness-related behaviors, helplessness, fatigue, and pain at study entry. Helplessness was more strongly associated with functioning in the Hispanics than in the African American or white patients. Pain was strongly associated with physical but not mental health. The models were quite robust, accounting for 41% to 68% of the variance for the two summary measures. Patients' attitudes toward their disease, fatigue, and pain have greater impact on self-perceived functional levels, as measured by the SF-36, than do more objective measures of disease activity and damage such as the presence of specific autoantibodies and/or the occurrence of specific organ involvement. Interventions designed to improve outcome may need to include ethnic-specific as well as general strategies.


A Functional Slow Recycling Pathway of Transferrin is Required for Growth of Chlamydia.
An inhibitor of host cell lysophospholipid acyltransferase, an enzyme involved in lipid metabolism blocked growth of the obligate intracellular pathogen Chlamydia through its action on the transport of transferrin (Tf) via the slow pathway of recycling. A detailed characterization of this inhibition revealed that Tf accumulated in vesicles positive for Rab11, with a concomitant reduction in the level of Tf found within the transport intermediate Rab4/11 hybrid vesicles. The net result was the failure to be recycled to the plasma membrane. In chlamydiae-infected cells, the Tf-containing Rab11-positive vesicles were typically found intimately associated with the inclusion, and treatment with the inhibitor caused their accumulation, suggesting that the timely progression and completion of Tf recycling was necessary for proper chlamydial growth. Growth inhibition by the compound could be negated by the simple removal of the Tf-containing fraction of the serum, a further indication that accumulation of Tf around the chlamydial inclusion was deleterious to the pathogen. Thus, it appears that manipulating the slow recycling pathway can have biological consequences for Chlamydia and implies the need to regulate carefully the interaction of the inclusion with this host trafficking pathway.


Climatic Impact of Volcanic Eruptions.
Volcanic eruptions have the potential to force global climate, provided they are explosive enough to emit at least 1-5 megaton of sulfur gases into the stratosphere. The sulfuric acid produced during oxidation of these gases will both absorb and reflect incoming solar radiation, thus warming the stratosphere and cooling the Earth's surface. Maximum global cooling on the order of 0.2-0.3°C, using instrumental temperature records, occurs in the first 2 years after the eruption, with lesser cooling possibly up to the 4th year. Equatorial eruptions are able to affect global climate, whereas mid- to high-latitude events will impact the hemisphere of origin. However, regional responses may differ, including the possibility of winter warming following certain eruptions. Also, El Niño warming may override the cooling induced by volcanic activity. Evaluation of different style eruptions as well as of multiple eruptions closely spaced in time beyond the instrumental record is attained through the analysis of ice-core, tree-ring, and geologic records. Using these data in conjunction with climate proxy data indicates that multiple eruptions may force climate on decadal time scales, as appears to have occurred during the Little Ice Age (i.e., roughly AD 1400s-1800s). The Toba mega-eruption of ~75,000 years ago may have injected extremely large amounts of material into the stratosphere that remained aloft for up to about 7 years. This scenario could lead to the initiation of feedback mechanisms within the climate system, such as cooling of sea-surface temperatures. These interacting mechanisms following a mega-eruption may cool climate on centennial time scales.


Efficacy of geoengineering to limit 21st century sea-level rise.
Geoengineering has been proposed as a feasible way of mitigating anthropogenic climate change, especially increasing global temperatures in the 21st century. The two main geoengineering options are limiting incoming solar radiation, or modifying the carbon cycle. Here we examine the impact of five geoengineering approaches on sea level; SO(2) aerosol injection into the stratosphere, mirrors in space, afforestation, biochar, and bioenergy with carbon sequestration. Sea level responds mainly at centennial time scales to temperature change, and has been largely driven by anthropogenic forcing since 1850. Making use a model of sea-level rise as a function of time-varying climate forcing factors (solar radiation, volcanism, and greenhouse gas emissions) we find that sea-level rise by 2100 will likely be 30 cm higher than 2000 levels despite all but the most aggressive geoengineering under all except the most stringent greenhouse gas emissions scenarios. The least risky and most desirable way of limiting sea-level rise is bioenergy with carbon sequestration. However aerosol injection or a space mirror system reducing insolation at an accelerating rate of 1 W m(-2) per decade from now to 2100 could limit or reduce sea levels. Aerosol injection delivering a constant 4 W m(-2) reduction in radiative forcing (similar to a 1991 Pinatubo eruption every 18 months) could delay sea-level rise by 40-80 years. Aerosol injection appears to fail cost-benefit analysis unless it can be maintained continuously, and damage caused by the climate response to the aerosols is less than about 0.6% Global World Product.


Health-related quality of life in patients with schizophrenia during treatment with long-acting, injectable risperidone.
We investigated the impact of treatment with long-acting, injectable risperidone versus placebo on health-related quality of life (HRQoL) in patients with schizophrenia. Results are discussed in the context of HRQoL in the general U.S. population. Patients with DSM-IV schizophrenia entered a randomized, double-blind, placebo-controlled trial. After screening, previous antipsychotics were discontinued, and oral risperidone was titrated up to a dose of 4 mg/day over 1 week. Patients were then randomly assigned to receive placebo [N = 92] or long-acting risperidone (25 [N = 93], 50 [N = 97], or 75 mg [N = 87] every 2 weeks) for 12 weeks. HRQoL was measured using the Medical Outcomes Study Short-Form 36-item questionnaire (SF-36). At week 12, patients receiving long-acting risperidone had improved significantly (p <.05) in 5 domains of the SF-36 (bodily pain, general health, social functioning, role-emotional, and mental health) compared with patients receiving placebo. The effect was greatest for the 25-mg group, with significant improvement versus placebo in 6 domains (p <.05). At baseline, all SF-36 domain scores except bodily pain were significantly lower (p <.05) than normal values in all groups. With placebo, scores in all 8 domains remained below normal values after 12 weeks, while patients receiving long-acting risperidone showed improvement in HRQoL toward normal levels, with clinically meaningful improvements in all mental-health domains. In the 25-mg group, scores in 7 domains were not statistically different from normal values after 12 weeks. Long-acting, injectable risperidone improved HRQoL toward normal levels. After 12 weeks, HRQoL of patients receiving 25 mg was not significantly different from normal.


Histamine exerts multiple effects on expression of genes associated with epidermal barrier function.
The role of epidermal barrier genes in the pathogenesis of atopic skin inflammation has recently been highlighted. Cytokines that are abundant in the skin during inflammation have been shown to exert various effects on the expression of barrier genes, although the role of histamine in this area of skin biology is not yet fully understood. To assess the effect of stimulation with histamine on keratinocytes by analysis of the pathways involved in epidermal barrier integrity. We performed a gene expression analysis of histamine-stimulated keratinocytes. Functional changes were tested using the dye penetration assay. Differential changes in filaggrin and the filaggrin-processing enzyme bleomycin hydrolase (BLMH) were validated at the protein level, and expression was also assessed in filaggrin knock-down keratinocytes. Histamine altered expression of multiple barrier genes. Expression of filaggrin was downregulated, as was that of other markers, thus suggesting the presence of delayed/aberrant keratinocyte differentiation. Expression of genes involved in cellular adhesiveness and genes of protease expression was dysregulated, but expression of protease inhibitors was increased. BLMH was upregulated in keratinocytes subjected to histamine and filaggrin knockdown. Histamine exerts a dual effect on epidermal barrier genes; it suppresses keratinocyte differentiation and dysregulates genes of cellular adhesiveness, although it induces genes contributing to stratum corneum function. Upregulation of BLMH and protease inhibitors could support maintenance of the permeability barrier by enhanced generation of moisturizing compounds and suppressed desquamation. In contrast, in the case of stratum corneum damage, histamine could enhance transcutaneous sensitization.


Evaluation of phacoemulsification-induced oxidative stress and damage of cultured human corneal endothelial cells in different solutions using redox fluorometry microscopy.
To describe the basic concept of redox fluorometry microscopy and investigate its efficacy in evaluating the state of cultured human corneal endothelial cells in different solutions when ultrasonic energy was applied in vitro. Human corneal endothelial cells from human donor tissue not suitable for transplantation were cultured. A phacoemulsification probe with a 30° round, 1.1-mm TurboSonics(®) ABS™ Tip (Alcon, Fort Worth, Texas) was introduced into culture dishes filled with balanced salt solution (BSS) and BSS plus (Alcon, Fort Worth, Texas). Cellular autofluorescence images were obtained using a Zeiss inverted microscope. The redox fluorometric ratio, which can be related to cellular metabolism, mitochondrial distribution patterns, which can shift in reaction to environmental changes, and cell size were analysed with a software program. Human corneal endothelial cells exposed to increasing phacoemulsification times and ultrasonic energy displayed dose-dependent decreases in redox ratios. At a lower ultrasonic power and time, BSS plus showed significantly less change in redox ratio than BSS and control (p < 0.05, Mann-Whitney test). As ultrasonic power and time increased, BSS plus had no more significance. Redox fluorometry, with further technological improvement, might be an interesting and potentially useful tool for evaluation of phacoemulsification-induced corneal endothelial damage and screening of protective agents in vitro.


Utilization of Tissue Ploidy Level Variation in de Novo Transcriptome Assembly of Pinus sylvestris.
Compared to angiosperms, gymnosperms lag behind in the availability of assembled and annotated genomes. Most genomic analyses in gymnosperms, especially conifer tree species, rely on the use of de novo assembled transcriptomes. However, the level of allelic redundancy and transcript fragmentation in these assembled transcriptomes, and their effect on downstream applications have not been fully investigated. Here, we assessed three assembly strategies for short-reads data, including the utility of haploid megagametophyte tissue during de novo assembly as single-allele guides, for six individuals and five different tissues in Pinus sylvestris We then contrasted haploid and diploid tissue genotype calls obtained from the assembled transcriptomes to evaluate the extent of paralog mapping. The use of the haploid tissue during assembly increased its completeness without reducing the number of assembled transcripts. Our results suggest that current strategies that rely on available genomic resources as guidance to minimize allelic redundancy are less effective than the application of strategies that cluster redundant assembled transcripts. The strategy yielding the lowest levels of allelic redundancy among the assembled transcriptomes assessed here was the generation of SuperTranscripts with Lace followed by CD-HIT clustering. However, we still observed some levels of heterozygosity (multiple gene fragments per transcript reflecting allelic redundancy) in this assembled transcriptome on the haploid tissue, indicating that further filtering is required before using these assemblies for downstream applications. We discuss the influence of allelic redundancy when these reference transcriptomes are used to select regions for probe design of exome capture baits and for estimation of population genetic diversity.


Expression of Drosophila epidermal growth factor receptor homologue in mitotic cell populations.
Expression of the Drosophila homologue (DER) of the human epidermal growth factor (EGF) receptor has been studied during development by RNA blot hybridizations and in situ hybridizations. One of the hypothetical functions of the protein encoded by this gene is mitotic signal transduction and, therefore, we have searched for evidence of its expression in mitotic cell populations. Increased DER transcript levels were detected in virtually all cells in cellular blastoderm embryos, indicative of the onset of transcription of the gene at this stage of development. These transcripts persist at least until the formation of the ventral furrow at the beginning of gastrulation. Expression of DER has been investigated in populations of nondividing cells, imaginal cells and nervous tissue in third instar larvae. By RNA blot hybridizations, we have shown that imaginal discs contain the bulk of the DER transcripts present in the whole third instar larvae. This result has been confirmed by in situ hybridizations. All imaginal discs, representing dividing cells, contain DER transcripts while salivary glands, representing nondividing cells, do not. Within the central nervous system, hybridization of the DER probe is confined to small clusters of cells in the brain cortex. During oogenesis, cell division programs are highly ordered temporally and spatially. Follicle cells express high levels of DER RNA during their mitotic phase of growth but lose these transcripts as they cease division. Nurse cells also express DER transcripts at lower levels even though they are not dividing. However, nurse cell transcripts represent stored maternal RNA species destined for use during preblastoderm embryonic development. These results indicate that the DER gene is expressed in mitotic cell populations during several stages of development and is not expressed in certain populations of nonmitotic cells.


In conditions of limited chromophore supply rods entrap 11-cis-retinal leading to loss of cone function and cell death.
RPE65 is a retinoid isomerase required for the production of 11-cis-retinal, the chromophore of both cone and rod visual pigments. We recently established an R91W knock-in mouse strain as homologous animal model for patients afflicted by this mutation in RPE65. These mice have impaired vision and can only synthesize minute amounts of 11-cis-retinal. Here, we investigated the consequences of this chromophore insufficiency on cone function and pathophysiology. We found that the R91W mutation caused cone opsin mislocalization and progressive geographic cone atrophy. Remnant visual function was mostly mediated by rods. Ablation of rod opsin corrected the localization of cone opsin and improved cone retinal function. Thus, our analyses indicate that under conditions of limited chromophore supply rods and cones compete for 11-cis-retinal that derives from regeneration pathway(s) which are reliant on RPE65. Due to their higher number and the instability of cone opsin, rods are privileged under this condition while cones suffer chromophore deficiency and degenerate. These findings reinforce the notion that in patients any effective gene therapy with RPE65 needs to target the cone-rich macula directly to locally restore the cones' chromophore supply outside the reach of rods.


Data-Oriented Constitutive Modeling of Plasticity in Metals.
Constitutive models for plastic deformation of metals are typically based on flow rules determining the transition from elastic to plastic response of a material as function of the applied mechanical load. These flow rules are commonly formulated as a yield function, based on the equivalent stress and the yield strength of the material, and its derivatives. In this work, a novel mathematical formulation is developed that allows the efficient use of machine learning algorithms describing the elastic-plastic deformation of a solid under arbitrary mechanical loads and that can replace the standard yield functions with more flexible algorithms. By exploiting basic physical principles of elastic-plastic deformation, the dimensionality of the problem is reduced without loss of generality. The data-oriented approach inherently offers a great flexibility to handle different kinds of material anisotropy without the need for explicitly calculating a large number of model parameters. The applicability of this formulation in finite element analysis is demonstrated, and the results are compared to formulations based on Hill-like anisotropic plasticity as reference model. In future applications, the machine learning algorithm can be trained by hybrid experimental and numerical data, as for example obtained from fundamental micromechanical simulations based on crystal plasticity models. In this way, data-oriented constitutive modeling will also provide a new way to homogenize numerical results in a scale-bridging approach.


Breakthrough mucormycosis after voriconazole use in a case of invasive fungal rhinosinusitis due to Curvularia lunata.
Invasive fungal rhinosinusitis (FRS) is a potentially fatal illness requiring early diagnosis and aggressive treatment with surgery and antifungals. We report a case of chronic FRS in a recently diagnosed diabetic individual due to Curvularia lunata. Imaging revealed extension into the right orbit and right basifrontal lobe. This was further complicated by development of nosocomial mucormycosis which was attributed to voriconazole therapy. The patient responded well to debridement and amphotericin B based therapy. To our knowledge, there are no reported cases of invasive FRS due to Curvularia lunata. Also, breakthrough mucormycosis on voriconazole therapy is rarely seen in non-malignancy, non-transplant settings. The possibility of rare fungal infections (community and nosocomial) should be entertained in developing settings where fungal spores are ubiquitous.


Turbine sound may influence the metamorphosis behaviour of estuarine crab megalopae.
It is now widely accepted that a shift towards renewable energy production is needed in order to avoid further anthropogenically induced climate change. The ocean provides a largely untapped source of renewable energy. As a result, harvesting electrical power from the wind and tides has sparked immense government and commercial interest but with relatively little detailed understanding of the potential environmental impacts. This study investigated how the sound emitted from an underwater tidal turbine and an offshore wind turbine would influence the settlement and metamorphosis of the pelagic larvae of estuarine brachyuran crabs which are ubiquitous in most coastal habitats. In a laboratory experiment the median time to metamorphosis (TTM) for the megalopae of the crabs Austrohelice crassa and Hemigrapsus crenulatus was significantly increased by at least 18 h when exposed to either tidal turbine or sea-based wind turbine sound, compared to silent control treatments. Contrastingly, when either species were subjected to natural habitat sound, observed median TTM decreased by approximately 21-31% compared to silent control treatments, 38-47% compared to tidal turbine sound treatments, and 46-60% compared to wind turbine sound treatments. A lack of difference in median TTM in A. crassa between two different source levels of tidal turbine sound suggests the frequency composition of turbine sound is more relevant in explaining such responses rather than sound intensity. These results show that estuarine mudflat sound mediates natural metamorphosis behaviour in two common species of estuarine crabs, and that exposure to continuous turbine sound interferes with this natural process. These results raise concerns about the potential ecological impacts of sound generated by renewable energy generation systems placed in the nearshore environment.


The IHF regulon of exponentially growing Pseudomonas putida cells.
Integration host factor (IHF) sites are largely absent from intergenic regions of ORFs encoding central metabolic functions in Pseudomonas putida mt-2. To gain an insight into this unequal distribution of otherwise abundant IHF-binding sequences, the transcriptome of IHF-plus and IHF-minus cells growing exponentially on glucose as sole carbon source was examined. In parallel, the cognate metabolic fluxes of the wild-type P. putida strain and its ihfA derivative were determined by culturing cells to a steady-state physiological regime with (13)C-labelled glucose. While expression of many transcripts was altered by the lack of IHF, flux balance analysis revealed that the ihfA mutation did not influence central carbon metabolism. Identification of multiple IHF sites adjacent to genes responsive to the factor allowed a refinement of the consensus and the mapping of the preferred binding positions for activation or repression of associated promoters. That few (if any) of the genes affected by IHF involved core pathways suggested that the central carbon metabolism tolerates the loss of the factor. Instead, IHF controlled various cell surface-related functions and downregulated genes encoding ribosomal proteins, the alpha subunit of RNA polymerase and components of the ATP synthase. These results were confirmed with lacZ fusions to a suite of promoters detected in the transcriptome as affected by IHF. Taken together, the data suggest that IHF plays a role in the physiological shift that sets P. putida for entering stationary phase.


Mesenchymal stem cells: characteristics and clinical applications.
Mesenchymal stem cells (MSCs) are bone marrow populating cells, different from hematopoietic stem cells, which possess an extensive proliferative potential and ability to differentiate into various cell types, including: osteocytes, adipocytes, chondrocytes, myocytes, cardiomyocytes and neurons. MSCs play a key role in the maintenance of bone marrow homeostasis and regulate the maturation of both hematopoietic and non-hematopoietic cells. The cells are characterized by the expression of numerous surface antigens, but none of them appears to be exclusively expressed on MSCs. Apart from bone marrow, MSCs are located in other tissues, like: adipose tissue, peripheral blood, cord blood, liver and fetal tissues. MSCs have been shown to be powerful tools in gene therapies, and can be effectively transduced with viral vectors containing a therapeutic gene, as well as with cDNA for specific proteins, expression of which is desired in a patient. Due to such characteristics, the number of clinical trials based on the use of MSCs increase. These cells have been successfully employed in graft versus host disease (GvHD) treatment, heart regeneration after infarct, cartilage and bone repair, skin wounds healing, neuronal regeneration and many others. Of special importance is their use in the treatment of osteogenesis imperfecta (OI), which appeared to be the only reasonable therapeutic strategy. MSCs seem to represent a future powerful tool in regenerative medicine, therefore they are particularly important in medical research.


Effects of intima stiffness and plaque morphology on peak cap stress.
Rupture of the cap of a vulnerable plaque present in a coronary vessel may cause myocardial infarction and death. Cap rupture occurs when the peak cap stress exceeds the cap strength. The mechanical stress within a cap depends on the plaque morphology and the material characteristics of the plaque components. A parametric study was conducted to assess the effect of intima stiffness and plaque morphology on peak cap stress. Models with idealized geometries based on histology images of human coronary arteries were generated by varying geometric plaque features. The constructed multi-layer models contained adventitia, media, intima, and necrotic core sections. For adventitia and media layers, anisotropic hyperelastic material models were used. For necrotic core and intima sections, isotropic hyperelastic material models were employed. Three different intima stiffness values were used to cover the wide range reported in literature. According to the intima stiffness, the models were classified as stiff, intermediate and soft intima models. Finite element method was used to compute peak cap stress. The intima stiffness was an essential determinant of cap stresses. The computed peak cap stresses for the soft intima models were much lower than for stiff and intermediate intima models. Intima stiffness also affected the influence of morphological parameters on cap stresses. For the stiff and intermediate intima models, the cap thickness and necrotic core thickness were the most important determinants of cap stresses. The peak cap stress increased three-fold when the cap thickness was reduced from 0.25 mm to 0.05 mm for both stiff and intermediate intima models. Doubling the thickness of the necrotic core elevated the peak cap stress by 60% for the stiff intima models and by 90% for the intermediate intima models. Two-fold increase in the intima thickness behind the necrotic core reduced the peak cap stress by approximately 25% for both intima models. For the soft intima models, cap thickness was less critical and changed the peak cap stress by 55%. However, the necrotic core thickness was more influential and changed the peak cap stress by 100%. The necrotic core angle emerged as a critical determinant of cap stresses where a larger angle lowered the cap stresses. Contrary to the stiff and intermediate intima models, a thicker intima behind the necrotic core increased the peak cap stress by approximately 25% for the soft intima models. Adventitia thickness and local media regression had limited effects for all three intima models. For the stiff and intermediate intima models, the cap thickness was the most important morphological risk factor. However for soft intima models, the necrotic core thickness and necrotic core angle had a bigger impact on the peak cap stress. We therefore need to enhance our knowledge of intima material properties if we want to derive critical morphological plaque features for risk evaluation.


Why is evaluation of the cost effectiveness of audit so difficult? The example of thrombolysis for suspected acute myocardial infarction.
Cost effectiveness analysis is an established technique for evaluation of delivery of health care, but its use to evaluate clinical audit is rarely reported. Thrombolysis for suspected acute myocardial infarction is a commonly used therapy of established effectiveness and an appropriate subject for audit in many healthcare settings. To measure the cost effectiveness of audit of thrombolysis in some district general hospitals. Cost of audit per extra patient treated with thrombolysis (incremental cost effectiveness ratio). Prospective agreement with physicians to undertake repeated audits of a specific aspect of the management of patients with acute myocardial infarction. Baseline measurement of the proportion of these patients given thrombolysis in each hospital were made, as were three subsequent retrospective audits, giving time series of measurements. Costs were estimated from records of staff time and other resources used in each hospital; effectiveness was estimated by fitting the results to a model which assumed a uniform rate of increase over time in the proportion of eligible patients given thrombolysis which might be accelerated by regular audit. Upper and lower limits for main outcome measure were derived from sensitivity analysis of costs and logistic regression of time series data. Five district general hospitals in North West Thames Regional Health Authority including one control hospital were used, starting in April 1991 when widespread medical audit was first introduced. Between the first and last audits, the proportion of patients with suspected acute myocardial infarctions receiving thrombolysis rose in three of the hospitals undertaking audit by 20% to 37% and fell by 6% in the fourth (although this hospital started with a rate in excess of 90%). The corresponding change in the control hospital was an increase of 15%. The differences between each of the auditing hospitals and control hospital were not significant, except in one case, where 51 extra treatments per year were attributable to audit (95% confidence intervals (95% CIs) 0.5 to 61 cases per year). Estimated overall costs in each hospital ranged from 3700 Pounds to 5200 Pounds for data collection, a series of four audit meetings, and subsequent actions. The central estimate of cost effectiveness in the three responsive hospitals ranged from 101 Pounds to 392 Pounds per extra case given thrombolysis, with very wide 95% CIs. In the fourth hospital audit had zero effectiveness as defined in this study. Methodological difficulties were encountered which need to be considered in future economic evaluations of clinical audit and related activities. These were: (a) adequate control for other factors influencing clinical behaviour; (b) uncertainties about the sustainability of changes in behaviour associated with audit; and (c) the relative infrequency in a single hospital of specific clinical events leading to small numbers for analysis. These difficulties constitute major challenges for the economic evaluation of clinical audit. They are most likely to be overcome in a large study which compares clinical audit with other interventions aiming for the same quality improvement, such as patient specific reminders or educational programmes.


Is there proofreading during polypeptide synthesis?
The stoichiometric efficiency with which ternary complexes containing Phe-tRNAphe and Leu-tRNAleu support polypeptide synthesis has been compared in a poly(U)-directed, steady-state translation system. When unfractionated tRNA is used to support synthesis, the number of discharged ternary complexes per peptide bond formed is an average of 48 times greater for leucine than for phenylalanine. When three purified leucine isoacceptor species are tested, they each show a characteristic ratio of ternary complexes discharged per missense insertion, normalized to that for phenylalanine: these are 103, 76, and 45 for Leu- tRNA2leu , Leu- tRNA3leu , and Leu- tRNA4leu , respectively. The data are consistent with the functioning of a proofreading mechanism during translation.


PERB11 (MIC): a polymorphic MHC gene is expressed in skin and single nucleotide polymorphisms are associated with psoriasis.
The susceptibility genes for psoriasis remain to be identified. At least one of these must be in the major histocompatibility complex (MHC) to explain associations with alleles at human leucocyte antigen (HLA)-A, -B, -C, -DR, -DQ and C4. In fact, most of these alleles are components of just two ancestral haplotypes (AHs) designated 13.1 and 57.1. Although relevant MHC gene(s) could be within a region of at least 4 Mb, most studies have favoured the area near HLA-B and -C. This region contains a large number of non-HLA genes, many of which are duplicated and polymorphic. Members of one such gene family, PERB11.1 and PERB11.2, are expressed in the skin and are encoded in the region between tumour necrosis factor and HLA-B. To investigate the relationship of PERB11.1 alleles to psoriasis, sequence based typing was performed on 97 patients classified according to age of onset and family history. The frequency of the PERB11.1*06 allele is 44% in type I psoriasis but only 7% in controls (Pc = 0.003 by Fisher's exact test, two-tailed). The major determinant of this association is a single nucleotide polymorphism (SNP) within intron 4. In normal and affected skin, expression of PERB11 is mainly in the basal layer of the epidermis including ducts and follicles. PERB11 is also present in the upper keratin layers but there is relative deficiency in the intermediate layers. These findings suggest a possible role for PERB11 and other MHC genes in the pathogenesis of psoriasis.


Decreased risk of acute myocardial infarction in stroke patients receiving acupuncture treatment: a nationwide matched retrospective cohort study.
Whether acupuncture protects stroke patients from acute myocardial infarction (AMI) has not been studied previously. The purpose of this study was to investigate the risk of AMI among stroke patients receiving acupuncture treatment. Taiwan's National Health Insurance Research Database was used to conduct a retrospective cohort study of 23475 stroke patients aged 40-79 years receiving acupuncture treatment and 46950 propensity score-matched stroke patients not receiving acupuncture treatment who served as controls from 2000 to 2004. Both stroke cohorts were followed until the end of 2009 and were adjusted for immortal time to measure the incidence and adjusted hazard ratios (HRs) with 95 % confidence intervals (CIs) for new-onset AMI in multivariate Cox proportional hazard models. Stroke patients who received acupuncture treatment (9.2 per 1000 person-years) exhibited a lower incidence of AMI compared with those who did not receive acupuncture treatment (10.8 per 1000 person-years), with an HR of 0.86 (95 % CI, 0.80-0.93) after adjusting for age, sex, low income, coexisting medical conditions and medications. The relationship between acupuncture treatment and AMI risk was investigated in female stroke patients (HR, 0.85; 95 % CI, 0.76-0.95), male stroke patients (HR, 0.87; 95 % CI, 0.80-0.95), patients from 50 to 59 years of age (HR, 0.75; 95 % CI, 0.63-0.90), patients from 60 to 69 years of age (HR, 0.85; 95 % CI, 0.75-0.95), patients suffering from ischemic stroke (HR, 0.87; 95 % CI, 0.79-0.95), and patients suffering from hemorrhagic stroke (HR, 0.62; 95 % CI, 0.44-0.88). We raised the possibility that acupuncture may be effective in lowering the risk of AMI in stroke patients aged 50-69 in this study, which was limited by a lack of information regarding stroke severity and acupuncture points. Our results suggest that prospective randomized trials are needed to establish the efficacy of acupuncture in preventing AMI.


Measuring impact in the Millennium Development Goal era and beyond: a new approach to large-scale effectiveness evaluations.
Evaluation of large-scale programmes and initiatives aimed at improvement of health in countries of low and middle income needs a new approach. Traditional designs, which compare areas with and without a given programme, are no longer relevant at a time when many programmes are being scaled up in virtually every district in the world. We propose an evolution in evaluation design, a national platform approach that: uses the district as the unit of design and analysis; is based on continuous monitoring of different levels of indicators; gathers additional data before, during, and after the period to be assessed by multiple methods; uses several analytical techniques to deal with various data gaps and biases; and includes interim and summative evaluation analyses. This new approach will promote country ownership, transparency, and donor coordination while providing a rigorous comparison of the cost-effectiveness of different scale-up approaches.


How memory generates heterogeneous dynamics in temporal networks.
Empirical temporal networks display strong heterogeneities in their dynamics, which profoundly affect processes taking place on these networks, such as rumor and epidemic spreading. Despite the recent wealth of data on temporal networks, little work has been devoted to the understanding of how such heterogeneities can emerge from microscopic mechanisms at the level of nodes and links. Here we show that long-term memory effects are present in the creation and disappearance of links in empirical networks. We thus consider a simple generative modeling framework for temporal networks able to incorporate these memory mechanisms. This allows us to study separately the role of each of these mechanisms in the emergence of heterogeneous network dynamics. In particular, we show analytically and numerically how heterogeneous distributions of contact durations, of intercontact durations, and of numbers of contacts per link emerge. We also study the individual effect of heterogeneities on dynamical processes, such as the paradigmatic susceptible-infected epidemic spreading model. Our results confirm in particular the crucial role of the distributions of intercontact durations and of the numbers of contacts per link.


Efficiency of the four proteasome subtypes to degrade ubiquitinated or oxidized proteins.
The proteasome is responsible for selective degradation of proteins. It exists in mammalian cells under four main subtypes, which differ by the combination of their catalytic subunits: the standard proteasome (β1-β2-β5), the immunoproteasome (β1i-β2i-β5i) and the two intermediate proteasomes (β1-β2-β5i and β1i-β2-β5i). The efficiency of the four proteasome subtypes to degrade ubiquitinated or oxidized proteins remains unclear. Using cells expressing exclusively one proteasome subtype, we observed that ubiquitinated p21 and c--myc were degraded at similar rates, indicating that the four 26S proteasomes degrade ubiquitinated proteins equally well. Under oxidative stress, we observed a partial dissociation of 26S into 20S proteasomes, which can degrade non-ubiquitinated oxidized proteins. Oxidized calmodulin and hemoglobin were best degraded in vitro by the three β5i-containing 20S proteasomes, while their native forms were not degraded. Circular dichroism analyses indicated that ubiquitin-independent recognition of oxidized proteins by 20S proteasomes was triggered by the disruption of their structure. Accordingly, β5i-containing 20S proteasomes degraded unoxidized naturally disordered protein tau, while 26S proteasomes did not. Our results suggest that the three β5i-containing 20S proteasomes, namely the immunoproteasome and the two intermediate proteasomes, might help cells to eliminate proteins containing disordered domains, including those induced by oxidative stress.


α-Lipoic acid protects diabetic apolipoprotein E-deficient mice from nephropathy.
Both hyperglycemia and hyperlipidemia increase oxidative stress and contribute to the development of diabetic nephropathy (DN). We investigated the effects of α-lipoic acid, a natural antioxidant and a cofactor in the multienzyme complexes, on the development of DN in diabetic apolipoprotein E-deficient mice. Twelve-week-old male apoE-/- mice on C57BL/6J genetic background were made diabetic with injections of streptozotocin (STZ). STZ-treated diabetic apoE-/- mice and non-diabetic control were fed with a synthetic high-fat (HF) diet with or without lipoic acid (LA) supplementation. Multiple parameters including plasma glucose, cholesterol, oxidative stress markers, cytokines, and kidney cortex gene expression, and glomerular morphology were evaluated. LA supplementation markedly protected the β cells, reduced cholesterol levels, and attenuated albuminuria and glomerular mesangial expansion in the diabetic mice. Renoprotection by LA was equally effective regardless of whether the dietary supplementation was started 4 weeks before, simultaneously with, or 4 weeks after the induction of diabetes by STZ. LA supplementation significantly improved DN and oxidative stress in the diabetic mice. Severity of albuminuria was positively correlated with level of thiobarbituric acid reactive substances (TBARs) in the kidney (r(2)=0.62, P<.05). Diabetes significantly changed the kidney expression of Rage, Sod2, Tgfb1 and Ctgf, Pdp2, nephrin, and Lias. LA supplementation corrected these changes except that it further suppressed the expression of the Lias gene coding for lipoic acid synthase. Our data indicate that LA supplementation effectively attenuates the development and progression of DN through its antioxidant effect as well as enhances glucose oxidation.


Hepatic binding and internalization of low density lipoprotein-gold conjugates in rats treated with 17 alpha-ethinylestradiol.
Receptor-mediated hepatic uptake of low density lipoproteins (LDL) conjugated to colloidal gold was studied by perfusion of livers from rats treated for 5 d with 17 alpha-ethinylestradiol. Estrogen treatment resulted in a marked decrease in serum lipid and lipoprotein concentrations. After 15 min of perfusion the conjugate was bound to the hepatic microvilli of both control and estrogen-treated rats; the estrogen-treated rats showed an 8- to 11-fold greater number of membrane-bound conjugates. The conjugates were bound to the membrane receptor by the LDL particle because the gold granules were regularly displaced from the membrane by 20 +/- 3.2 nm, the diameter of LDL. Internalization of the conjugate, evident by gold particles in multivesicular bodies, occurred at coated pits at the base of the microvillus where coated vesicles containing a single gold-LDL conjugate were released. After 1 h of perfusion, the livers from the estrogen-treated rats showed all phases of endocytosis and incorporation into multivesicular bodies of the conjugate. After 2 h of perfusion, there was congregation of gold-labeled lysosomes near the bile canaliculi. Gold-LDL conjugates were also observed to bind and be internalized by Kupffer cells and sinusoidal endothelium. These findings indicate that estrogen treatment induces hepatic receptors for LDL. The catabolic pathway of binding and endocytosis of the conjugate is similar to that seen in fibroblasts, although slower. Because gold-LDL conjugates were also present in the Kupffer and endothelial cells, the uptake of LDL by the liver involves the participation of more than a single cell type.


Microbial contamination of contact lens cases in the west of Scotland.
A cross-sectional study of 178 asymptomatic contact lens wearers attending 10 contact lens practices in the west of Scotland was conducted over a 4 month period. The aims of the study were to identify specific microbial contaminants in lens cases, to determine the rate of contamination of such containers and to assess the value of the steps involved in different lens care regimens in the prevention of case contamination. Microbial contamination affected 53% of lens cases. Cases used with conventional wear and disposable systems were contaminated at similar rates and, therefore, the advantage of regular lens replacement may have been lost if these lenses were stored in contaminated cases. Four percent of lens cases were contaminated with amoebal species and all of these showed concomitant bacterial colonisation. These findings imply that case hygiene is probably as important as lens hygiene if new or disinfected lenses are not to be immediately re-contaminated by storage in dirty cases. Unfortunately simple and effective methods of lens and case disinfection, which would be suitable for use in the average home environment, are not yet available. It follows that frequent and regular disposal of lens cases may prove to be a necessary measure to prevent the build-up of microbial colonisation in such containers.


The effect of insertional suspensory branch desmitis on racing performance in juvenile Thoroughbred racehorses.
Limited information exists regarding the prognosis for juvenile racehorses sustaining injury to the suspensory ligament branch insertion (JISBI). To investigate the effect of JISBI on racing performance; and to assess whether the severity of JISBI is prognostically important. Retrospective cohort study. Records were reviewed for 896 juvenile Thoroughbreds in training, to identify horses with JISBI limited to one branch (cases). The ability of cases to start a race was compared to their cohorts. Race records of maternal siblings were compared to cases. The prevalence of JISBI for this population was 9.5%. Cohorts were 3.2 times more likely to start as a 2-year-old and 3.6 times more likely to start as a 3-year-old compared to horses with JISBI. Of 58 cases that were compared to maternal siblings, the total earnings per start (EPS), 2- and 3-year-old EPS were all reduced (P<0.01). Cases raced a mean of 3.8 times compared to 7.4 races for controls over the study period and were 113.2 days older by the time of first start (P<0.01). 'Speed figures' and EPS were, however, similar between cases that had raced and controls. Cases with a moderate-severe lesion (≥ Grade 2) were at significantly greater risk of reinjury than mild cases (P<0.05). JISBI causes decreased racing ability as a 2-year-old. Mild cases perform similarly to controls by their 3-year-old season, but more severe cases demonstrate reduced ability as a 3-year-old, with an increased likelihood of reinjury. Although the wastage was higher in case horses with JISBI, individual cases that make it to a race have similar earnings per start as their controls.


The protective effects of oral low-dose quercetin on diabetic nephropathy in hypercholesterolemic mice.
Diabetic nephropathy (DN) is one of the most important causes of chronic renal disease, and the incidence of DN is increasing worldwide. Considering our previous report (Gomes et al., 2014) indicating that chronic treatment with oral low-dose quercetin (10 mg/Kg) demonstrated anti-oxidative, anti-apoptotic and renoprotective effects in the C57BL/6J model of DN, we investigated whether this flavonoid could also have beneficial effects in concurrent DN and spontaneous atherosclerosis using the apolipoprotein E-deficient mouse (apoE(-/-)). Streptozotocin was used to induce diabetes (100 mg/kg/day, 3 days) in male apoE(-/-) mice (8 week-old). After 6 weeks, the mice were randomly separated into DQ: diabetic apoE(-/-) mice treated with quercetin (10 mg/kg/day, 4 weeks, n = 8), DV: diabetic ApoE(-/-) mice treated with vehicle (n = 8) and ND: non-treated non-diabetic mice (n = 8). Quercetin treatment diminished polyuria (~30%; p < 0.05), glycemia (~25%, p < 0.05), normalized the hypertriglyceridemia. Moreover, this bioflavonoid diminished creatininemia (~30%, p < 0.01) and reduced proteinuria but not to normal levels. We also observed protective effects on the renal structural changes, including normalization of the index of glomerulosclerosis and kidney weight/body weight. Our data revealed that quercetin treatment significantly reduced DN in hypercholesterolemic mice by inducing biochemical changes (decrease in glucose and triglycerides serum levels) and reduction of glomerulosclerosis. Thus, this study highlights the relevance of quercetin as an alternative therapeutic option for DN, including in diabetes associated with dyslipidemia.


The effect of proton pump inhibitors on the human microbiota.
Proton pump inhibitors (PPIs) are commonly used to treat acid-related diseases, most notably gastroesophageal reflux disease. PPIs are designed to shut down the gastric proton pump (H+/K+-ATPase) of parietal cells, thereby raising the pH of the stomach. While effective, a number of side effects have been associated with PPI use. Naturally occurring bacteria, some of which are acid-producing and contain ATPase enzymes, have also been found within the stomach, upper gastrointestinal tract, and oral cavity. Likewise, a number of fungi are known to inhabit the human body; some of these fungi contain H+-ATPase enzymes. Recent literature has suggested that PPIs may be inadvertently affecting these bacteria and fungi in two different ways: 1) PPIs may directly target the proton pumps of the bacteria and fungi, and/or 2) PPIs may indirectly affect the microenvironment of the flora via changes in pH. These unintended interactions are exasperated by the systemic distribution of PPIs throughout the body and may potentially lead to some of the side effects observed with PPI use. Herein we summarize what is currently known about the interactions between the PPIs and the natural human microbiota.


Suppression of cell-mediated immunocompetence after subchronic exposure to diethylstilbestrol in female B6C3F1 mice.
The effect of the nonsteroidal estrogenic compound, diethylstilbestrol (DES) on cell-mediated immunocompetence in the mouse has been assessed. Adult female B6C3F1 mice were injected (s.c.) with 0.2, 1.0 and 4.0 mg/kg of DES daily for 14 days. All immunological tests and toxicological parameters were determined at least 24 hr after the last exposure. Thymic involution and hepatomegaly were noted at the lowest dose of DES. The most sensitive indicator of immunosuppression by DES was the delayed hypersensitivity response (DHR) to keyhole limpet hemocyanin in mice sensitized without adjuvant. The lowest dose of DES produced a 93% decrease, compared with a 39% decrease for the same DHR model in mice sensitized to keyhole limpet hemocyanin plus adjuvant and a 63% decrease for the DHR to sheep erythrocytes. A 35% decrease of the acute inflammatory response to carrageenin was produced by the lowest dose of DES. Day 2 proliferative responses to both concanavalin A and phytohemagglutinin (T-cell mitogens) were depressed by the lowest dose of DES, whereas significant suppression of the response to lipopolysaccharide (B-cell mitogen) was only noted at the highest dose of DES. The effects of DES on Day 3 proliferative responses were less dramatic. The mixed lymphocyte response was significantly suppressed by 1.0 and 4.0 mg/kg of DES on all days in culture. The reversibility of the DES effects was studied by resting the mice for 30 days between exposure and measuring a given parameter. All effects on organ weights and the depression of both the sheep erythrocytes DHR and the carrageenin inflammatory response were reversed.


Association between drug and vaccine use and acute immune thrombocytopenia in childhood: a case-control study in Italy.
Immune thrombocytopenic purpura (ITP) is an immunomediated disease characterized by a decrease in platelet count and, in its more severe forms, by bleeding symptoms. Many drugs have been implicated in the pathogenesis of drug-induced thrombocytopenia in adults; only limited data on drug-related ITP in children have been published. Our study was set up to evaluate the consistency of the association between drug and vaccine use and ITP in children. This study is part of an Italian multicentre study on adverse drug reactions in children, coordinated by the Italian National Institute of Health, which was started in November 1999 and is ongoing. The study was conducted by enrolling all children aged more than 1 month who were hospitalized through the paediatric emergency department for the following conditions: thrombocytopenia (platelet count <100 x 103/L); acute neurological disorders; non-infectious mucocutaneous diseases and vasculitis; and endoscopically confirmed gastroduodenal lesions and/or clinically defined haematemesis and melaena. Children with chronic pathologies or concomitant diagnoses of cancer or immunodeficiency were not included in our study. During hospital admission, a physician interviewed parents using a structured questionnaire. The main aim of the interview was to collect information on drug exposure in a time period of 3 weeks and vaccine exposure in a period of 6 weeks preceding hospitalization. Using a case-control study design, exposure of children with thrombocytopenia (cases) to drugs and vaccines was compared with similar exposure of children with gastroduodenal lesions and neurological disorders (controls); this allowed us to estimate the odds ratios (ORs) of the occurrence of thrombocytopenia associated with the use of drugs or vaccines. Up to December 2007, the study population included 387 cases of thrombocytopenia and 1924 controls. Despite the low platelet count, ITP was generally a mild disease, without serious bleeding in the majority of cases and associated with a short length of hospital stay. After adjusting for concurrent use of other drugs, use of the antibacterials was associated with a more than 2-fold increase in the risk of developing ITP (OR 2.4; 95% CI 1.8, 3.1). Mucolytics and NSAIDs were associated with an OR of 1.9; 95% CI 1.2, 2.9 and 1.5; 95% CI 1.0, 2.1 respectively, while paracetamol (acetaminophen) was associated with an OR of 1.5; 95% CI 1.2, 2.0. MMR vaccination was associated with an increased risk of developing ITP (OR 2.4; 95% CI 1.2, 4.7). The results of this study provide evidence for an association between ITP and exposure to selected antibacterials, NSAIDs, paracetamol, mucolytics and MMR vaccination.


In vivo analgesic, antipyretic, and anti-inflammatory potential in Swiss albino mice and in vitro thrombolytic activity of hydroalcoholic extract from Litsea glutinosa leaves.
The study was conducted to evaluate the in vitro thrombolytic activity, and in vivo analgesic, anti-inflammatory and antipyretic potentials of different hydrocarbon soluble extracts of Litsea glutinosa leaves for the first time widely used in the folkloric treatments in Bangladesh. This work aimed to create new insights on the fundamental mechanisms of the plant extracts involved in these activities. In thrombolytic activity assay, a significant clot disruption was observed at dose of 1 mg/mL for each of the extracts (volume 100 μL) when compared to the standard drug streptokinase. The n-hexane, ethyl acetate, chloroform, and crude methanolic extracts showed 32.23 ± 0.26, 37.67 ± 1.31, 43.13 ± 0.85, and 46.78 ± 0.9% clot lysis, respectively, whereas the positive control streptokinase showed 93.35 ± 0.35% disruption at the dose of 30,000 I.U. In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (15.54 ± 0.37 sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug ketorolac (16.38 ± 0.27 sec). In acetic acid induced writhing test, the crude methanolic extract showed significant (P <0.01 and P <0.001) analgesic potential at doses 250 and 500 mg/kg body weight (45.98 and 56.32% inhibition, respectively), where ketorolac showed 64.36% inhibition. In anti-inflammatory activity test, the crude methanolic extract showed significant (P <0.001) potential at doses 250 and 500 mg/kg body weight (1.51 ± 0.04 and 1.47 ± 0.03 mm paw edema, respectively), where ketorolac showed 1.64 ± 0.05 mm edema after 3 h of carrageenan injection. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (32.78 ± 0.46°C) at dose of 500 mg/kg-body weight, when the standard (at dose 150 mg/kg-body weight) exerted 33.32 ± 0.67°C temperature after 3 h of administration. Our results yield that the crude hydroalcoholic extract has better effects than the other in all trials. In the context, it can be said that the leaves of L. glutinosa possess remarkable pharmacological effects, and justify its traditional use as analgesic, antipyretic, anti-inflammatory, and thrombolytic agent.


Cardiac involvement in hemochromatosis.
Cardiac hemochromatosis or primary iron-overload cardiomyopathy is an important and potentially preventable cause of heart failure. This is initially characterized by diastolic dysfunction and arrhythmias and in later stages by dilated cardiomyopathy. Diagnosis of iron overload is established by elevated transferrin saturation (>55%) and elevated serum ferritin (>300 ng/mL). Genetic testing for mutations in the HFE (high iron) gene and other proteins, such as hemojuvelin, transferrin receptor, and ferroportin, should be performed if secondary causes of iron overload are ruled out. Patients should undergo comprehensive 2D and Doppler echocardiography to evaluate their systolic and diastolic function. Newer modalities like strain imaging and speckle-tracking echocardiography hold promise for earlier detection of cardiac involvement. Cardiac magnetic resonance imaging with measurement of T2* relaxation times can help quantify myocardial iron overload. In addition to its value in diagnosis of cardiac iron overload, response to iron reduction therapy can be assessed by serial imaging. Therapeutic phlebotomy and iron chelation are the cornerstones of therapy. The average survival is less than a year in untreated patients with severe cardiac impairment. However, if treated early and aggressively, the survival rate approaches that of the regular heart failure population.


The Ankrd2 protein, a link between the sarcomere and the nucleus in skeletal muscle.
Ankrd2 may be a link between the sarcomere and the nucleus; a similar role has recently been proposed for CARP that has a high level of structural and functional conservation with Ankrd2. Both Ankrd2 and CARP are involved in striated muscle hypertrophy. The mechanism by which muscle stretch is sensed and signals are transduced is still unknown; however, Ankrd2 and CARP could play similar roles in pathways leading to hypertrophy, the triggering mechanisms being heart pressure overload monitored by CARP and mechanical stretch in skeletal muscle monitored by Ankrd2. Recently Ankrd2 and CARP have been proposed as members of a family of muscle ankyrin repeat proteins (MARPs) that form a complex with titin, myopalladin and calpain protease p94, involved in signaling and regulation of gene expression in response to muscle stress. Here, we show that Ankrd2 is able to interact with the Z-disc protein telethonin as well as being able to interact with three transcription factors: YB-1, PML and p53. Ankrd2 binding to the ubiquitous transcription factor YB-1 can be demonstrated both in vitro and in vivo; this is not very surprising, since a similar interaction was previously described for CARP. However, the interactions with PML and p53 are unexpected new findings, with interesting implications in the Ankrd2 signaling cascade. Ankrd2 co-localizes with the transcriptional co-activator and co-repressor PML in nuclear bodies (NBs) in human myoblasts as detected by confocal immunofluorescence. Interestingly, we show that Ankrd2 not only binds the tumor suppressor protein p53 both in vitro and in vivo but also enhances the up-regulation of the p21(WAFI/CIPI) promoter by p53. Therefore, our findings strengthen the hypothesis that Ankrd2 may be involved in sensing stress signals and linking these to muscle gene regulation.


Outcome Trajectories among Homeless Individuals with Mental Disorders in a Multisite Randomised Controlled Trial of Housing First.
Housing First (HF) has been shown to improve housing stability, on average, for formerly homeless adults with mental illness. However, little is known about patterns of change and characteristics that predict different outcome trajectories over time. This article reports on latent trajectories of housing stability among 2140 participants (84% followed 24 months) of a multisite randomised controlled trial of HF. Data were analyzed using generalised growth mixture modeling for the total cohort. Predictor variables were chosen based on the original program logic model and detailed reviews of other qualitative and quantitative findings. Treatment group assignment and level of need at baseline were included in the model. In total, 73% of HF participants and 43% of treatment-as-usual (TAU) participants were in stable housing after 24 months of follow-up. Six trajectories of housing stability were identified for each of the HF and TAU groups. Variables that distinguished different trajectories included gender, age, prior month income, Aboriginal status, total time homeless, previous hospitalizations, overall health, psychiatric symptoms, and comorbidity, while others such as education, diagnosis, and substance use problems did not. While the observed patterns and their predictors are of interest for further research and general service planning, no set of variables is yet known that can accurately predict the likelihood of particular individuals benefiting from HF programs at the outset.


Vinegar Production from Jabuticaba (Myrciaria jaboticaba) Fruit Using Immobilized Acetic Acid Bacteria.
Cell immobilization comprises the retention of metabolically active cells inside a polymeric matrix. In this study, the production of jabuticaba (Myrciaria jaboticaba) vinegar using immobilized Acetobacter aceti and Gluconobacter oxydans cells is proposed as a new method to prevent losses of jabuticaba fruit surplus. The pulp of jabuticaba was processed and Saccharomyces cerevisiae CCMA 0200 was used to ferment the must for jabuticaba wine production. Sugars, alcohols (ethanol and glycerol) and organic acids were assayed by high-performance liquid chromatography. Volatile compounds were determined by gas chromatography-flame ionization detector. The ethanol content of the produced jabuticaba wine was approx. 74.8 g/L (9.5% by volume) after 168 h of fermentation. Acetic acid fermentation for vinegar production was performed using a mixed culture of immobilized A. aceti CCT 0190 and G. oxydans CCMA 0350 cells. The acetic acid yield was 74.4% and productivity was 0.29 g/(L·h). The vinegar had particularly high concentrations of citric (6.67 g/L), malic (7.02 g/L) and succinic (5.60 g/L) acids. These organic acids give a suitable taste and flavour to the vinegar. Seventeen compounds (aldehydes, higher alcohols, terpene, acetate, diether, furans, acids, ketones and ethyl esters) were identified in the jabuticaba vinegar. In conclusion, vinegar was successfully produced from jabuticaba fruits using yeast and immobilized mixed cultures of A. aceti and G. oxydans. To the best of our knowledge, this is the first study to use mixed culture of immobilized cells for the production of jabuticaba vinegar.


Competing Factor Models of Child and Adolescent Psychopathology.
Co-occurring psychological disorders are highly prevalent among children and adolescents. To date, the most widely utilised factor model used to explain this co-occurrence is the two factor model of internalising and externalising (Achenbach 1966). Several competing models of general psychopathology have since been reported as alternatives, including a recent three factor model of Distress, Fear and Externalising Dimensions (Krueger 1999). Evidence for the three factor model suggests there are advantages to utilising a more complex model. Using the British Child and Adolescent Mental Health Survey 2004 data (B-CAMHS; N = 7997), confirmatory factor analysis was used to test competing factor structure models of child and adolescent psychopathology. The B-CAMHS was an epidemiological survey of children between the ages of 5 and 16 in Great Britain. Child psychological disorders were assessed using the Strength and Difficulties Questionnaire (Goodman 1997), and the Development and Wellbeing Assessment (Goodman et al. 2000). A range of covariates and risk variables including trauma, parent mental health and family functioning where subsequently utilised within a MIMIC model framework to predict each dimension of the 2 and three factor structure models. Two models demonstrated acceptable fit. The first complimented Achenbach's Internalising and Externalising structure. The three factor model was found to have highly comparable fit indices to the two factor model. The second order models did not accurately represent the data nor did an alternative three factor model of Internalising, Externalising and ADHD. The two factor and three factor MIMIC models observed unique profiles of risk for each dimension. The findings suggest that child and adolescent psychopathology may also be accurately conceptualised in terms of distress, fear and externalising dimensions. The MIMIC models demonstrated that the Distress and Fear dimensions have their own unique etiological profile of risk. This study directly informs future measurement models of child and adolescent psychopathology and demonstrates the effectiveness of a three factor model.


Mapping language networks and their association with verbal abilities in paediatric epilepsy using MEG and graph analysis.
Recent theoretical models of language have emphasised the importance of integration within distributed networks during language processing. This is particularly relevant to young patients with epilepsy, as the topology of the functional network and its dynamics may be altered by the disease, resulting in reorganisation of functional language networks. Thus, understanding connectivity within the language network in patients with epilepsy could provide valuable insights into healthy and pathological brain function, particularly when combined with clinical correlates. The objective of this study was to investigate interactions within the language network in a paediatric population of epilepsy patients using measures of MEG phase synchronisation and graph-theoretical analysis, and to examine their association with language abilities. Task dependent increases in connectivity were observed in fronto-temporal networks during verb generation across a group of 22 paediatric patients (9 males and 13 females; mean age 14 years). Differences in network connectivity were observed between patients with typical and atypical language representation and between patients with good and poor language abilities. In addition, node centrality in left frontal and temporal regions was significantly associated with language abilities, where patients with good language abilities had significantly higher node centrality within inferior frontal and superior temporal regions of the left hemisphere, compared to patients with poor language abilities. Our study is one of the first to apply task-based measures of MEG network synchronisation in paediatric epilepsy, and we propose that these measures of functional connectivity and node centrality could be used as tools to identify critical regions of the language network prior to epilepsy surgery.


Symptoms of depression as a risk factor for incident diabetes: findings from the National Health and Nutrition Examination Epidemiologic Follow-up Study, 1971-1992.
Symptoms of depression may predict incident diabetes independently or through established risk factors for diabetes. US men and women aged 25-74 years who were free of diabetes at baseline (n = 6,190) were followed from 1971 to 1992 (mean, 15.6 years; standard deviation, 6) for incident diabetes. Depressive symptoms were measured by using the General Well-Being Depression subscale and were categorized to compare persons with high (9%), intermediate (32%), and low (59%) numbers of symptoms. The incidence of diabetes was highest among participants reporting high numbers of depressive symptoms (7.3 per 1,000 person-years) and did not differ between persons reporting intermediate and low numbers of symptoms (3.4 and 3.6 per 1,000 person-years, respectively) (p < 0.01 for high vs. low). In the subset of participants with less than a high school education (a marker of low socioeconomic status), the risk of developing diabetes was three times higher (95% confidence interval: 2.0, 4.7) for persons reporting high versus low numbers of depressive symptoms. These results persisted following adjustment for established diabetes risk factors. Depressive symptoms had no impact on diabetes incidence among persons with at least a high school education. Results suggest an independent role for depressive symptoms in the development of diabetes in populations with low educational attainment.


β1-Integrin- and KV1.3 channel-dependent signaling stimulates glutamate release from Th17 cells.
Although the impact of Th17 cells on autoimmunity is undisputable, their pathogenic effector mechanism is still enigmatic. We discovered soluble N-ethylmaleimide-sensitive factor attachment receptor (SNARE) complex proteins in Th17 cells that enable a vesicular glutamate release pathway that induces local intracytoplasmic calcium release and subsequent damage in neurons. This pathway is glutamine dependent and triggered by binding of β1-integrin to vascular cell adhesion molecule 1 (VCAM-1) on neurons in the inflammatory context. Glutamate secretion could be blocked by inhibiting either glutaminase or KV1.3 channels, which are known to be linked to integrin expression and highly expressed on stimulated T cells. Although KV1.3 is not expressed in CNS tissue, intrathecal administration of a KV1.3 channel blocker or a glutaminase inhibitor ameliorated disability in experimental neuroinflammation. In humans, T cells from patients with multiple sclerosis secreted higher levels of glutamate, and cerebrospinal fluid glutamine levels were increased. Altogether, our findings demonstrate that β1-integrin- and KV1.3 channel-dependent signaling stimulates glutamate release from Th17 cells upon direct cell-cell contact between Th17 cells and neurons.


Automated white matter total lesion volume segmentation in diabetes.
WM lesion segmentation is often performed with the use of subjective rating scales because manual methods are laborious and tedious; however, automated methods are now available. We compared the performance of total lesion volume grading computed by use of an automated WM lesion segmentation algorithm with that of subjective rating scales and expert manual segmentation in a cohort of subjects with type 2 diabetes. Structural T1 and FLAIR MR imaging data from 50 subjects with diabetes (age, 67.7 ± 7.2 years) and 50 nondiabetic sibling pairs (age, 67.5 ± 9.4 years) were evaluated in an institutional review board-approved study. WM lesion segmentation maps and total lesion volume were generated for each subject by means of the Statistical Parametric Mapping (SPM8) Lesion Segmentation Toolbox. Subjective WM lesion grade was determined by means of a 0-9 rating scale by 2 readers. Ground-truth total lesion volume was determined by means of manual segmentation by experienced readers. Correlation analyses compared manual segmentation total lesion volume with automated and subjective evaluation methods. Correlation between average lesion segmentation and ground-truth total lesion volume was 0.84. Maximum correlation between the Lesion Segmentation Toolbox and ground-truth total lesion volume (ρ = 0.87) occurred at the segmentation threshold of k = 0.25, whereas maximum correlation between subjective lesion segmentation and the Lesion Segmentation Toolbox (ρ = 0.73) occurred at k = 0.15. The difference between the 2 correlation estimates with ground-truth was not statistically significant. The lower segmentation threshold (0.15 versus 0.25) suggests that subjective raters overestimate WM lesion burden. We validate the Lesion Segmentation Toolbox for determining total lesion volume in diabetes-enriched populations and compare it with a common subjective WM lesion rating scale. The Lesion Segmentation Toolbox is a readily available substitute for subjective WM lesion scoring in studies of diabetes and other populations with changes of leukoaraiosis.