Identification of FOXH1 mutations in patients with sporadic conotruncal heart defect.
Conotruncal heart defects (CTD) are an important subtype of congenital heart disease that occur due to abnormality in the development of the cardiac outflow tract (OFT). FOXH1 is a transcription factor that participates in the morphogenesis of the right ventricle and OFT. In this study, we confirmed the expression of FOXH1 in mouse and human embryos during OFT development. We also scanned the coding exons and splicing regions of the FOXH1 gene in 605 patients with sporadic CTD and 300 unaffected controls, from which we identified seven heterozygous FOXH1 gene mutations. According to bioinformatics analysis results, they were predicted potentially deleterious at conserved amino acid sites. Western blot was used to show that all the variants decreased the expression of FOXH1 protein, while dual-luciferase reporter assay showed that six of them, with an exception of p.P35R, had enhanced abilities to modulate the expression of MEF2C, which interacts with NKX2.5 and is involved in cardiac growth. The electrophoretic mobility shift assays result showed that two mutations altered DNA-binding abilities of mutant FOXH1 proteins. Phenotype heterogeneity was found in patients with the same mutation. These results indicate that FOXH1 mutations lead to disease-causing functional changes that contribute to the occurrence of CTD.


How to: EUCAST recommendations on the screening procedure E.Def 10.1 for the detection of azole resistance in Aspergillus fumigatus isolates using four-well azole-containing agar plates.
The emergence of azole-resistant Aspergillus fumigatus isolates is a matter of significant concern in Europe, with countries reporting resistance rates (which can be as high as 30%) in hospitalized patients. Consequently, the treatment guidelines in The Netherlands, the country with the highest documented prevalence of azole-resistant A. fumigatus, has just been revised to now recommend initial therapy with combination therapy until the susceptibility pattern is known. Therefore, susceptibility testing of clinically relevant isolates has been strongly recommended in the ESCMID-EFISG aspergillosis guidelines. Furthermore, mixed azole-susceptible and azole-resistant (isogenic as well as non-isogenic) infections have been reported to occur, which implies that colonies of clinical cultures may harbour various phenotypes of azole susceptibility. The EUCAST-AFST (European Committee on Antimicrobial Susceptibility Testing Subcommittee on Antifungal Susceptibility Testing) has released a new screening method document (E.Def 10.1) for the detection of azole-resistant A. fumigatus isolates and updated the QC tables for antifungal susceptibility testing with associated QC endpoints. This review described in detail how to perform the screening test. This "How to document" is based on the EUCAST azole agar screening method document E.Def 10.1 and the QC tables for antifungal susceptibility testing document, v 2.0 (available at http://www.eucast.org/ast_of_fungi/qcafsttables/) CONTENTS: The method is based on the inoculation of azole-containing and azole-free agars and visual determination of fungal growth after one and two days of incubation. It can easily be implemented in routine laboratories of clinical microbiology and has been validated for simultaneous testing of up to five A. fumigatus colonies using itraconazole and voriconazole (mandatory), and posaconazole (optional). This easy-to-use screening procedure for the detection of azole resistance in clinical A. fumigatus isolates will allow rapid testing in the daily routine of the microbiology laboratory and thus facilitate earlier appropriate therapy.


SLP-2 interacts with Parkin in mitochondria and prevents mitochondrial dysfunction in Parkin-deficient human iPSC-derived neurons and Drosophila.
Mutations in the Parkin gene (PARK2) have been linked to a recessive form of Parkinson's disease (PD) characterized by the loss of dopaminergic neurons in the substantia nigra. Deficiencies of mitochondrial respiratory chain complex I activity have been observed in the substantia nigra of PD patients, and loss of Parkin results in the reduction of complex I activity shown in various cell and animal models. Using co-immunoprecipitation and proximity ligation assays on endogenous proteins, we demonstrate that Parkin interacts with mitochondrial Stomatin-like protein 2 (SLP-2), which also binds the mitochondrial lipid cardiolipin and functions in the assembly of respiratory chain proteins. SH-SY5Y cells with a stable knockdown of Parkin or SLP-2, as well as induced pluripotent stem cell-derived neurons from Parkin mutation carriers, showed decreased complex I activity and altered mitochondrial network morphology. Importantly, induced expression of SLP-2 corrected for these mitochondrial alterations caused by reduced Parkin function in these cells. In-vivo Drosophila studies showed a genetic interaction of Parkin and SLP-2, and further, tissue-specific or global overexpression of SLP-2 transgenes rescued parkin mutant phenotypes, in particular loss of dopaminergic neurons, mitochondrial network structure, reduced ATP production, and flight and motor dysfunction. The physical and genetic interaction between Parkin and SLP-2 and the compensatory potential of SLP-2 suggest a functional epistatic relationship to Parkin and a protective role of SLP-2 in neurons. This finding places further emphasis on the significance of Parkin for the maintenance of mitochondrial function in neurons and provides a novel target for therapeutic strategies.


Development of an Evidence-Based Practical Diagnostic Checklist and Corresponding Clinical Exam for Low Back Pain.
The purpose of this study was to use scientific evidence to develop a practical diagnostic checklist and corresponding clinical exam for patients presenting with low back pain (LBP). An iterative process was conducted to develop a diagnostic checklist and clinical exam for LBP using evidence-based diagnostic criteria. The checklist and exam were informed by a systematic review focused on summarizing current research evidence for office-based clinical evaluation of common conditions causing LBP. Diagnostic categories contained within the checklist and exam include nociceptive pain, neuropathic pain, and sensitization. Nociceptive pain subcategories include discogenic, myofascial, sacroiliac, and zygapophyseal (facet) joint pain. Neuropathic pain categories include neurogenic claudication, radicular pain, radiculopathy, and peripheral entrapment (piriformis and thoracolumbar syndrome). Sensitization contains 2 subtypes, central and peripheral sensitization. The diagnostic checklist contains individual diagnostic categories containing evidence-based criteria, applicable examination procedures, and checkboxes to record clinical findings. The checklist organizes and displays evidence for or against a working diagnosis. The checklist may help to ensure needed information is obtained from a patient interview and exam in a variety of primary spine care settings (eg, medical, chiropractic). The available evidence informs reasonable working diagnoses for many conditions causing or contributing to LBP. A practical diagnostic process including an exam and checklist is offered to guide clinical evaluation and demonstrate evidence for working diagnoses in clinical settings.


The power of imagination--how anticipatory mental imagery alters perceptual processing of fearful facial expressions.
Expectancies strongly shape our perception of the world and preconceptions about stimulus characteristics can even bias the sensory system for illusory percepts. Here we assessed with functional magnetic resonance imaging how anticipatory mental imagery of a mildly fearful face created a predictive bias that proactively altered perception of highly fearful faces and generated the "illusion" of reduced fearfulness. We found that anticipatory activation of the fusiform gyrus (FG) was modulated by the fearfulness of the imagined face. Further during anticipatory imagery, regulatory influences from the lateral and ventromedial prefrontal cortex on the FG primed the perceptual system for a subsequent misperception. This was achieved by increasing perceptual activation in higher-order brain regions for the evaluation of affective valence and contextual framing, while at the same time restricting bottom-up arousal and attention to fearful expressions. Anticipatory mental imagery may thus represent an effective antecedent strategy through which emotional perception can be significantly altered.


From behavioural analyses to models of collective motion in fish schools.
Fish schooling is a phenomenon of long-lasting interest in ethology and ecology, widely spread across taxa and ecological contexts, and has attracted much interest from statistical physics and theoretical biology as a case of self-organized behaviour. One topic of intense interest is the search of specific behavioural mechanisms at stake at the individual level and from which the school properties emerges. This is fundamental for understanding how selective pressure acting at the individual level promotes adaptive properties of schools and in trying to disambiguate functional properties from non-adaptive epiphenomena. Decades of studies on collective motion by means of individual-based modelling have allowed a qualitative understanding of the self-organization processes leading to collective properties at school level, and provided an insight into the behavioural mechanisms that result in coordinated motion. Here, we emphasize a set of paradigmatic modelling assumptions whose validity remains unclear, both from a behavioural point of view and in terms of quantitative agreement between model outcome and empirical data. We advocate for a specific and biologically oriented re-examination of these assumptions through experimental-based behavioural analysis and modelling.


Specialty differences in the management of asthma. A cross-sectional assessment of allergists' patients and generalists' patients in a large HMO.
To examine the differences in medical management and quality of life between patients with asthma who receive their primary asthma care from allergists and those who receive their care from generalists in a large health maintenance organization (HMO). We conducted a cross-sectional study of patients with asthma in a large HMO (Kaiser Permanente, Northwest Region, Portland, Ore). Participants were 392 individuals aged 15 through 55 years with physician-diagnosed asthma, taking antiasthma medications, reporting current asthma symptoms, and receiving asthma care from an allergist or from a generalist. Primary outcomes include characteristics of asthma, health care utilization, and quality of life. Patients cared for by allergists tended to have more severe asthma than those cared for by generalists (P < .01). The allergists' patients tended to be older (38.6 +/- 9.6 years vs 35.7 +/- 12.6 years, P < .01), more atopic (91% vs 78%, P < .01), and more likely to report perennial (rather than seasonal) asthma (26% vs 36%, P < .04) than the generalists' patients. Patients receiving their primary asthma care from an allergist were considerably more likely than generalists' patients to report using inhaled anti-inflammatory agents (P < .01), oral steroids (P < .01), and regular (daily) breathing medications to control their asthma (P < .01). Allergists' patients were more likely to have asthma exacerbations treated in a clinic setting rather than an emergency department (P < .01). Furthermore, allergists' patients reported significantly improved quality of life as measured by several dimensions of the SF-36 scale (physical functioning, role emotional, bodily pain, and general health; P < .05). These findings suggest that specialist care of asthma is of benefit for patients with asthma in a large HMO. Specifically, the allergists' patients conformed more closely to national asthma management guidelines and reported better quality of life than did the generalists' patients.


Onchocerciasis in Anambra State, Southeast Nigeria: endemicity and clinical manifestations.
A cross-sectional study was performed to determine the current endemicity of onchocerciasis in Ayamelum Local Council, Anambra State, Southeast Nigeria, where community-directed treatment with ivermectin has been implemented for over a decade. An estimate of the endemicity of onchocerciasis was obtained using the rapid assessment method in 894 subjects from 13 communities selected by multistage sampling. Dermatological and ocular manifestations were analysed and classified using standard criteria. Onchocerca volvulus nodules were recorded in 86 (9.6 ± 1.9%) of the subjects, and 186 (20.8 ± 3.7%) had one or more of the various classes of onchocercal skin diseases (OSD). Prevalence was dependent on age (p = 0.001), but not on sex (p = 0.31). There was a total absence of symptoms in the youngest age group and a low prevalence among subjects in their second decade of life. Pearson's correlation showed a strong positive correlation between nodular rate and prevalence of chronic papular onchodermatitis (r = 0.943) and a poor correlation with acute papular onchodermatitis (r = 0.259). Age-dependent analysis of various classes of OSDs showed that the rate of acute papular onchodermatitis increased with age up to the third decade of life and decreased steadily thereafter, while the chronic forms of OSD increased with age for both sexes. Infection was dependent on occupation and proximity of the village to the vector breeding sites. Generally low prevalence in the population and absence of symptoms among the youngest age group emphasise the success of the intervention, but the persistent occurrence of acute disease may suggest a shortfall due to low coverage or non-compliance with the mass chemotherapeutic regimen.


Factors Associated with Highly Pathogenic Avian Influenza H5N2 Infection on Table-Egg Layer Farms in the Midwestern United States, 2015.
A case-control study was conducted among commercial table-egg layer and pullet operations in Iowa and Nebraska, United States, to investigate potential risk factors for infection with highly pathogenic avian influenza (HPAI) H5N2. A questionnaire was developed and administered to 28 case farms and 31 control farms. Data were collected at the farm and barn levels, enabling two separate analyses to be performed-the first a farm-level comparison of case farms vs. control farms, and the second a barn-level comparison between case barns on case farms and control barns on control farms. Multivariable logistic regression models were fit using a forward-selection procedure. Key risk factors identified were farm location in an existing control zone, rendering and garbage trucks coming near barns, dead-bird disposal located near barns, and visits by a company service person. Variables associated with a decreased risk of infection included visitors changing clothing, cleaning and disinfecting a hard-surface barn entryway, and ceiling/eaves ventilation in barns.


Physics-Based Visual Characterization of Molecular Interaction Forces.
Molecular simulations are used in many areas of biotechnology, such as drug design and enzyme engineering. Despite the development of automatic computational protocols, analysis of molecular interactions is still a major aspect where human comprehension and intuition are key to accelerate, analyze, and propose modifications to the molecule of interest. Most visualization algorithms help the users by providing an accurate depiction of the spatial arrangement: the atoms involved in inter-molecular contacts. There are few tools that provide visual information on the forces governing molecular docking. However, these tools, commonly restricted to close interaction between atoms, do not consider whole simulation paths, long-range distances and, importantly, do not provide visual cues for a quick and intuitive comprehension of the energy functions (modeling intermolecular interactions) involved. In this paper, we propose visualizations designed to enable the characterization of interaction forces by taking into account several relevant variables such as molecule-ligand distance and the energy function, which is essential to understand binding affinities. We put emphasis on mapping molecular docking paths obtained from Molecular Dynamics or Monte Carlo simulations, and provide time-dependent visualizations for different energy components and particle resolutions: atoms, groups or residues. The presented visualizations have the potential to support domain experts in a more efficient drug or enzyme design process.


The disruption of hepatic cytochrome p450 reductase alters mouse lipid metabolism.
To elucidate the role of hepatic cytochrome P450 oxidoreductase (POR) in lipid metabolism, we characterized perturbations in lipid homeostasis in a mouse model deficient in liver POR. Using an integrative approach in which transcriptomics, lipidomics, and various bioinformatic algorithms were employed, a disruption in liver lipid mobilization, oxidation, and electron transport functions were identified. Analyzing the promoters of genes in these biological processes identified common binding motifs for nuclear receptors sensitive to lipid status, while Srebp-1c binding sites were only identified in genes involved in lipid metabolism. POR-null mice had drastic increases in hepatic lipid content (diacylglycerols, triacylglycerols, phosphatidylcholine, and cholesterol esters) and a specific enrichment in n-7 and n-9 monounsaturated fatty acids (FAs). It was found that while transporters involved in peroxisomal FA oxidation were induced, mitochondrial oxidation appeared to be more tightly controlled, supporting the increase in monounsaturated FAs. Genes coding for hepatic transporters were differentially expressed, where lipid uptake was induced and efflux repressed, indicating that in the absence of hepatic POR the liver serves as a lipid reservoir. Furthermore, while significant changes in intestinal gene expression were found in POR-deficient mice, only minor changes to plasma and intestinal lipid content were observed. Thus, while liver POR plays an important role regulating gene expression and lipid metabolism locally, the hepatic deficiency of this enzyme reverberates throughout the biological system and produces a coordinated response to the low levels of circulating cholesterol and bile.


Pulmonary cryptococcosis.
Pulmonary cryptococcosis is the consequence of the inhalation of Cryptococcus neoformans, an encapsulated yeast, from various environmental sources. It is commonly accepted that the acquisition of the disease occurs early in life and that the disease is mostly related to a reactivation from a pulmonary site in immunocompromised hosts such as patients infected with human immunodeficiency virus. Clinical and radiological presentations of the disease are nonspecific and are more severe in immunocompromised hosts with acquired immunodeficiency syndrome (AIDS). However, fulminant forms have also been reported in apparently immunocompetent hosts. C. neoformans has rarely been responsible for colonization of the respiratory tract; this usually occurs in immunocompetent hosts with preexisting pulmonary disease. Diagnosis of pneumonia is obtained by either analysis of bronchoalveolar lavage in AIDS patients or by the histology/mycological analysis of a pulmonary nodule. In any case, a careful work-up for diagnosing dissemination should always be performed. Antifungal treatment has to be given in most of the immunocompetent hosts and always in those with any kind of immunodeficiency with isolated pulmonary cryptococcosis.


Sexually transmitted disease and human immunodeficiency virus. Is everyone at risk and does everyone have to pay?
The pattern of disease within populations depends on the complex interaction between individuals. For example, the linking of individuals to form social and sexual networks affects disease incidence and prevalence. As described by Potterat, the formation of these networks results in various patterns of disease spread. However, phenomena that occur among individuals do not necessarily mirror those observed in groups or populations. The interactions that characterize the relationship between individual and group level phenomena were considered during this session of the Tenth International Meeting of the International Society for STD Research. By examining these interactions we hoped to address the following two questions: Are all individuals equally at risk for sexually transmitted diseases (STD) and infection with human immunodeficiency virus (HIV)? and Must the costs and benefits of interventions designed to prevent these outcomes be divided equally among all individuals to ensure the success of such interventions?


The immunogenetics of human infectious diseases.
Twin and adoptee studies have indicated that host genetic factors are major determinants of susceptibility to infectious disease in humans. Twin studies have also found high heritabilities for many humoral and cellular immune responses to pathogen antigens, with most of the genetic component mapping outside of the major histocompatibility complex. Candidate gene studies have implicated several immunogenetic polymorphisms in human infectious diseases. HLA variation has been associated with susceptibility or resistance to malaria, tuberculosis, leprosy, AIDS, and hepatitis virus persistence. Variation in the tumor necrosis factor gene promoter has also been associated with several infectious diseases. Chemokine receptor polymorphism affects both susceptibility ot HIV-1 infection and the rate of progression to AIDS. Inactivating mutations of the gamma-interferon receptor lead to increased susceptibility to typical mycobacteria and disseminated BCG infection in homozygous children. The active form of vitamin D has immunomodulatory effects, and allelic variants of the vitamin D receptor appear to be associated with differential susceptibility to several infectious diseases. NRAMP1, a macrophage gene identified by positional cloning of its murine homologue, has been implicated in susceptibility to tuberculosis in Africans. Whole genome linkage analysis of multi-case families is now being used to map and identify new loci affecting susceptibility to infectious diseases. It is likely that susceptibility to most microorganisms is determined by a large number of polymorphic genes, and identification of these should provide insights into protective and pathogenic mechanisms in infectious diseases.


Serum lipoprotein(a) in patients with diabetes mellitus.
To investigate subjects with different types of diabetes mellitus regarding their serum levels of lipoprotein(a). High serum Lp(a) concentration is associated with a high risk of coronary heart disease. Diabetic patients are prone to developing coronary heart disease. The subjects were 66 type I diabetic patients, 100 type II diabetic patients treated with diet alone or diet combined with oral hypoglycemic agents, and 46 insulin-requiring type II diabetic patients. Subjects were compared with 142 nondiabetic outpatients. Subjects with insulin-requiring type II diabetes mellitus were found to have an increase both in serum Lp(a) concentration and in prevalence of serum Lp(a) concentration > 30 mg/dl compared with the other groups of diabetic patients and nondiabetic control subjects. A nonsignificant increase in the prevalence of coronary heart disease was also found in insulin-requiring type II diabetic patients. The levels of serum concentrations of Lp(a) were not significantly related to the degree of glycemic control, duration of diabetes, presence of macrovascular disease, or intake of female hormone therapy. High levels of Lp(a) in this group of diabetic patients could not be explained by the presence of albuminuria. Insulin-requiring type II diabetic patients have high levels of Lp(a). Chronic hyperinsulinemia might be an eventual causal factor.


Dynamics of associative polymers.
Current progress in understanding the dynamics of associating polymers is reviewed, with examples including both ionic and hydrogen bonding associations. A particular emphasis is placed on quantification of the strength of the interaction that sets the association lifetime. Knowledge of the interaction energy and the number density of associating groups allows a rational understanding of the linear viscoelastic response of many associating polymers.


Biomechanical adaptation of porcine carotid vascular smooth muscle to hypo and hypertension in vitro.
Previous research in arterial remodeling in response to changes in blood pressure seldom included both hyper- and hypotension. To compare the effects of low and high pressure on arterial remodeling and vascular smooth muscle tone and performance, we have utilized an in vitro model. Porcine carotid arteries were cultured for 3 days at 30 and 170mmHg and compared to controls cultured at 100mmHg for 1 and 3 days. On the first and last day of culture, pressure-diameter and pressure-wall thickness curves were measured under normal smooth muscle tone using a high-resolution ultrasonic device. Last-day experiments included measurements where vascular smooth muscle was contracted or totally relaxed. From the data wall cross-sectional area, Hudetz elastic modulus and a contraction index related to the diameter reduction under normal smooth muscle tone were calculated. We found that although wall cross-sectional area (indicating wall mass) did not change much, Hudetz elastic modulus was significantly reduced in the 3-day hypotension group. Inspection of the wall contraction index suggests that this is due to a reduction in the vascular smooth muscle tone. Further, the peak of contraction index was found to be shifted to higher pressures in the 3-day 170mmHg group. We conclude that vascular smooth muscle performance adapts to both hypo- and hypertension at short time scales and can alter the biomechanics of the vascular wall in vitro.


Dexamethasone- cyclophosphamide pulse in collagen vascular diseases: An observation.
Treatment of collagen vascular diseases like systemic sclerosis, dermatomyositis, systemic lupus erythematosus (SLE) and even overlap syndromes has been difficult since long. Monumental success of dexamethasone-cyclophosphamide pulse (DCP) in pemphigus has prompted many a dermatologist to try it in other autoimmune diseases. DCP was given as per standard regimen for six to nine pulses. Immunosuppressives were given for 12-18 months in dermatomyositis, SLE, and overlap syndrome, and for 12 months in systemic sclerosis. Daily dose of steroid was tapered off gradually. The treatment resulted in 90% improvement in skin binding in systemic sclerosis, 80% in exertional dyspnea, 40% in dysphagia, but minimum improvement was seen in Raynauds and digital tip ulcerations. No improvement in pigmentation was noted. In SLE, malar rash cleared in 70%, joint pain in 80%, oral ulcerations reduced in 80%, fever in 98%, and photosensitivity improved in one-third of patients. In dermatomyositis, improvement in muscle tenderness was seen in 100%, improvement in proximal myopathy and heliotrope rash in 80%, and improvement of shawl sign was observed in 80% of the patients. Some flattening of Gottron papules and plaques was noted in some patients. Both overlap patients improved significantly. Out of 24 patients, three were lost to follow-up, one resorted to homeopathic medicine and two expired (one dermatomyositis, one SLE). Side effects like hypertension, hyperglycemia, pyoderma, fungal infections, obesity, psychosis, etc. were seen in 25-30% of patients. We conclude that DCP is relatively safe, effective as well as cheap compared to methylprednisolone pulse. Side effects are also less compared to daily regimen of steroids. We also observed that patients who reported early and put on pulse early responded better.


Initiation of forward gait with lateral occurrence of emotional stimuli: general findings and relevance for pedestrians crossing roads.
With reference to theoretical models regarding links between emotions and actions, the present study examined whether the lateral occurrence of an emotional stimulus influences spatial and temporal parameters of gait initiation in 18 younger and 18 older healthy adults. In order to simulate road-crossing hazard for pedestrians, slides of approaching cars were used and they were presented in counterbalanced order with threatening slides from the International Affective Picture System (IAPS) and control slides of safe walking areas. Each slide was presented on the left side of the participant once the first step was initiated. The results evidenced medio-lateral shifts to the left for the first step (right foot) and to the right for the second step (left foot). These shifts were both modulated by the slide contents in such a way that the resulting distance between the screen and the foot (right or left) was larger with the IAPS and traffic slides than with the control slides. The slides did not affect the base of support, step length, step velocity and time of double support. Advancing age influenced the subjective impact of the slides and gait characteristics, but did not modulate medio-lateral shifts. The data extend evidence of fast, emotional modulation of stepping, with theoretical and applied consequences.


Biological identifications through DNA barcodes.
Although much biological research depends upon species diagnoses, taxonomic expertise is collapsing. We are convinced that the sole prospect for a sustainable identification capability lies in the construction of systems that employ DNA sequences as taxon 'barcodes'. We establish that the mitochondrial gene cytochrome c oxidase I (COI) can serve as the core of a global bioidentification system for animals. First, we demonstrate that COI profiles, derived from the low-density sampling of higher taxonomic categories, ordinarily assign newly analysed taxa to the appropriate phylum or order. Second, we demonstrate that species-level assignments can be obtained by creating comprehensive COI profiles. A model COI profile, based upon the analysis of a single individual from each of 200 closely allied species of lepidopterans, was 100% successful in correctly identifying subsequent specimens. When fully developed, a COI identification system will provide a reliable, cost-effective and accessible solution to the current problem of species identification. Its assembly will also generate important new insights into the diversification of life and the rules of molecular evolution.


Accelerometry-measured physical activity and inflammation after gestational diabetes.
Gestational diabetes mellitus (GDM) is associated with adverse metabolic outcomes after delivery. Physical activity practice improves the inflammatory profile; however, whether this association exists in women with prior GDM remains unknown. Our objective was to examine the cardiometabolic and inflammatory risk factors associated with accelerometer-based measures of physical activity in women with prior GDM. Ninety-six women who had GDM between 2003 and 2010 were tested 2.9 ± 2.2 yr after delivery. The physical activity practice was measured with ActiGraph GT3X (ActiGraph™, Pensacola, FL) accelerometers worn ≥ 5 d, and the time spent weekly in moderate to vigorous physical activity (MVPA) was derived. The waist circumference was measured and the inflammatory marker or cytokine concentrations were measured in fasting plasma by the xMAP technology using the Bio-Plex 200 system. The lipid profile was also measured from fasting blood samples. Only 31% of women accumulated at least 150 min of MVPA per week. No association was observed between the MVPA practice and any of the metabolic measurements in the whole group of women. The MVPA did not differ in groups stratified by waist circumference <88 or ≥ 88 cm. In women with waist circumference <88 cm, the MVPA was negatively correlated with circulating concentrations of C-reactive protein (r = -0.51, P = 0.006), leptin (r = -0.40, P = 0.008), plasminogen activator inhibitor-1 (r = -0.32, P = 0.04), and triglycerides (r = -0.44, P = 0.003). No association was seen with plasma interleukin-6; tumor necrosis factor-α; and total, LDL, or HDL cholesterol concentrations. These analyses suggest that in the years after delivery, longer time spent in MVPA practice is associated with a lower cardiometabolic risk only in women with prior GDM who do not have abdominal obesity.


Plant UVR8 photoreceptor senses UV-B by tryptophan-mediated disruption of cross-dimer salt bridges.
The recently identified plant photoreceptor UVR8 (UV RESISTANCE LOCUS 8) triggers regulatory changes in gene expression in response to ultraviolet-B (UV-B) light through an unknown mechanism. Here, crystallographic and solution structures of the UVR8 homodimer, together with mutagenesis and far-UV circular dichroism spectroscopy, reveal its mechanisms for UV-B perception and signal transduction. β-propeller subunits form a remarkable, tryptophan-dominated, dimer interface stitched together by a complex salt-bridge network. Salt-bridging arginines flank the excitonically coupled cross-dimer tryptophan "pyramid" responsible for UV-B sensing. Photoreception reversibly disrupts salt bridges, triggering dimer dissociation and signal initiation. Mutation of a single tryptophan to phenylalanine retunes the photoreceptor to detect UV-C wavelengths. Our analyses establish how UVR8 functions as a photoreceptor without a prosthetic chromophore to promote plant development and survival in sunlight.


Mechanical performance comparison of two surgical constructs for wrist four-corner arthrodesis via dorsal and radial approaches.
Four-corner arthrodesis, which involves fusing four carpal bones while removing the scaphoid bone, is a standard surgery for the treatment of advanced stages of wrist arthritis. Nowadays, it can be performed using a dorsal approach by fixing a plate to the bones and a new radial approach is in development. To date, there is no consensus on the biomechanically optimal and most reliable surgical construct for four-corner arthrodesis. To evaluate them biomechanically and thus assist the surgeon in choosing the best implant orientation, radial or dorsal, the two different four-corner arthrodesis surgical constructs were virtually simulated on a 3D finite element model representing all major structures of the wrist. Two different realistic load sets were applied to the model, representing common tasks for the elderly. Results consistency was assessed by comparing with the literature the force magnitude computed on the carpal bones. The Von Mises stress distribution in the radial and dorsal plates were calculated. Stress concentration was located at the plate-screw interface for both surgical constructs, with a maximum stress value of 413 MPa for the dorsal plate compared to 326 MPa for the radial plate, meaning that the stress levels are more unfavourable in the dorsal approach. Although some bending stress was found in one load case, the radial plate was mechanically more robust in the other load case. Despite some limitations, this study provides, for the first time, quantified evidence that the newly developed radial surgical construct is mechanically as efficient as the dorsal surgical construct.


Pluripotent stem cell heterogeneity and the evolving role of proteomic technologies in stem cell biology.
Stem cells represent obvious choices for regenerative medicine and are invaluable for studies of human development and drug testing. The proteomic landscape of pluripotent stem cells (PSCs), in particular, is not yet clearly defined; consequently, this field of research would greatly benefit from concerted efforts designed to better characterize these cells. In this concise review, we provide an overview of stem cell potency, highlight the types and practical implications of heterogeneity in PSCs and provide a detailed analysis of the current view of the pluripotent proteome in a unique resource for this rapidly evolving field. Our goal in this review is to provide specific insights into the current status of the known proteome of both mouse and human PSCs. This has been accomplished by integrating published data into a unified PSC proteome to facilitate the identification of proteins, which may be informative for the stem cell state as well as to reveal areas where our current view is limited. These analyses provide insight into the challenges faced in the proteomic analysis of PSCs and reveal one area--the cell surface subproteome--that would especially benefit from enhanced research efforts.


Do maladaptive attitudes cause depression?
The Dysfunctional Attitude Scale (DAS) is an inventory of beliefs about life. These beliefs attribute happiness to external events and reflect absolute expectations for one's own behavior and that of others. Such beliefs, according to some cognitive theorists, predispose persons to depression. To test this theory, we administered the DAS to private psychiatric outpatients. Thirty-five patients were tested when they were depressed and again when they were asymptomatic. Dysfunctional thinking was found to be more prominent during depression; this finding implies that the dysfunctional thinking of depressed persons is a symptom of their illness rather than a character trait. Next, we compared DAS scores for recovered depressives with scores for other stabilized psychiatric patients and for normal persons. Bipolar patients demonstrated less maladaptive thinking than all other groups. Only the bipolars showed significantly less maladaptive thinking than the major depressives.


Human papilloma virus-specific T cells can be generated from naïve T cells for use as an immunotherapeutic strategy for immunocompromised patients.
Human papilloma virus (HPV) is a known cause of cervical cancer, squamous cell carcinoma and laryngeal cancer. Although treatments exist for HPV-associated malignancies, patients unresponsive to these therapies have a poor prognosis. Recent findings from vaccine studies suggest that T-cell immunity is essential for disease control. Because Epstein-Barr Virus (EBV)-specific T cells have been highly successful in treating or preventing EBV-associated tumors, we hypothesized that the development of a manufacturing platform for HPV-specific T cells from healthy donors could be used in a third-party setting to treat patients with high-risk/relapsed HPV-associated cancers. Most protocols for generating virus-specific T cells require prior exposure of the donor to the targeted virus and, because the seroprevalence of high-risk HPV types varies greatly by age and ethnicity, manufacturing of donor-derived HPV-specific T cells has proven challenging. We, therefore, made systematic changes to our current Good Manufacturing Practice (GMP)-compliant protocols to improve antigen presentation, priming and expansion for the manufacture of high-efficacy HPV-specific T cells. Like others, we found that current methodologies fail to expand HPV-specific T cells from most healthy donors. By optimizing dendritic cell maturation and function with lipopolysaccharide (LPS) and interferon (IFN)γ, adding interleukin (IL)-21 during priming and depleting memory T cells, we achieved reliable expansion of T cells specific for oncoproteins E6 and E7 to clinically relevant amounts (mean, 578-fold expansion; n = 10), which were polyfunctional based on cytokine multiplex analysis. In the third-party setting, such HPV-specific T-cell products might serve as a potent salvage therapy for patients with HPV-associated diseases.


Copper deficiency-induced anemia is caused by a mitochondrial metabolic reprograming in erythropoietic cells.
The lack of copper has been associated with anemia, myelodysplastic syndromes and leukemia as well as with a loss in complex IV activity and an enlarged mitochondrial morphology. Mitochondria play a key role during the differentiation of hematopoietic stem cells by regulating the passage from a glycolytic to oxidative metabolism. The former is associated with cell proliferation and the latter with cell differentiation. Oxidative metabolism, which occurs inside mitochondria, is sustained by the respiratory chain, where complex IV is copper-dependent. We have hypothesized that a copper deficiency induces a mitochondrial metabolic reprogramming, favoring cell expansion over cell differentiation in erythropoiesis. Erythroid progression analysis of the bone marrow of mice fed with a copper deficient diet and of the in vitro erythropoiesis of human CD34+ cells treated with a bathocuproine - a copper chelator - showed a major expansion of progenitor cells and a decreased differentiation. Under copper deficiency, mitochondria switched to a higher membrane potential, lower oxygen consumption rate and lower ROS levels as compared with control cells. In addition, mitochondrial biomass was increased and an up-regulation of the mitochondrial fusion protein mitofusin 2 was observed. Most copper-deficient phenotypes were mimicked by the pharmacological inhibition of complex IV with azide. We concluded that copper deficiency induced a mitochondrial metabolic reprogramming, making hematopoietic stem cells favor progenitor cell expansion over cell differentiation.


Diagnostic use of computational retrotransposon detection: Successful definition of pathogenetic mechanism in a ciliopathy phenotype.
Among more than 5,000 human monogenic disorders with known causative genes, transposable element insertion of a Long Interspersed Nuclear Element 1 (LINE1, L1) is known as the mechanistic basis in only 13 genetic conditions. Meckel-Gruber syndrome is a rare ciliopathy characterized by occipital encephalocele and cystic kidney disease. Here, we document a boy with occipital encephalocele, post-axial polydactyly, and multicystic renal disease. A medical exome analysis detected a heterozygous frameshift mutation, c.4582_4583delCG p.(Arg1528Serfs*17) in CC2D2A in the maternally derived allele. The further use of a dedicated bioinformatics algorithm for detecting retrotransposon insertions led to the detection of an L1 insertion affecting exon 7 in the paternally derived allele. The complete sequencing and sequence homology analysis of the inserted L1 element showed that the L1 element was classified as L1HS (L1 human specific) and that the element had intact open reading frames in the two L1-encoded proteins. This observation ranks Meckel-Gruber syndrome as only the 14th disorder to be caused by an L1 insertion among more than 5,000 known human genetic disorders. Although a transposable element detection algorithm is not included in the current best-practice next-generation sequencing analysis, the present observation illustrates the utility of such an algorithm, which would require modest computational time and resources. Whether the seemingly infrequent recognition of L1 insertion in the pathogenesis of human genetic diseases might simply reflect a lack of appropriate detection methods remains to be seen.


Development of the Standards of Reporting of Neurological Disorders (STROND) checklist: A guideline for the reporting of incidence and prevalence studies in neuroepidemiology.
Incidence and prevalence studies of neurologic disorders play an important role in assessing the burden of disease and planning services. However, the assessment of disease estimates is hindered by problems in reporting for such studies. Despite a growth in published reports, existing guidelines relate to analytical rather than descriptive epidemiologic studies. There are also no user-friendly tools (e.g., checklists) available for authors, editors, and peer reviewers to facilitate best practice in reporting of descriptive epidemiologic studies for most neurologic disorders. The Standards of Reporting of Neurological Disorders (STROND) is a guideline that consists of recommendations and a checklist to facilitate better reporting of published incidence and prevalence studies of neurologic disorders. A review of previously developed guidance was used to produce a list of items required for incidence and prevalence studies in neurology. A 3-round Delphi technique was used to identify the "basic minimum items" important for reporting, as well as some additional "ideal reporting items." An e-consultation process was then used in order to gauge opinion by external neuroepidemiologic experts on the appropriateness of the items included in the checklist. Of 38 candidate items, 15 items and accompanying recommendations were developed along with a user-friendly checklist. The introduction and use of the STROND checklist should lead to more consistent, transparent, and contextualized reporting of descriptive neuroepidemiologic studies resulting in more applicable and comparable findings and ultimately support better health care decisions.


Inhibition of CYP2B4 by 2-ethynylnaphthalene: evidence for the co-binding of substrate and inhibitor within the active site.
2-ethynylnaphthalene (2EN) is an effective mechanism-based inhibitor of CYP2B4. There are two inhibitory components: (1) irreversible inactivation of CYP2B4 (a typical time-dependent inactivation), and (2) a reversible component. The reversible component was unusual in that the degree of inhibition was not simply a characteristic of the enzyme-inhibitor interaction, but dependent on the size of the substrate molecule used to monitor residual activity. The effect of 2EN on the metabolism of seven CYP2B4 substrates showed that it was not an effective reversible inhibitor of substrates containing a single aromatic ring; substrates with two fused rings were competitively inhibited by 2EN; and larger substrates were non-competitively inhibited. Energy-based docking studies demonstrated that, with increasing substrate size, the energy of 2EN and substrate co-binding in the active site became unfavorable precisely at the point where 2EN became a competitive inhibitor. Hierarchical docking revealed potential allosteric inhibition sites separate from the substrate binding site.


C-peptide corrects endoneurial blood flow but not oxidative stress in type 1 BB/Wor rats.
Oxidative stress and neurovascular dysfunction have emerged as contributing factors to the development of experimental diabetic neuropathy (EDN) in streptozotocin-diabetic rodents. Additionally, depletion of C-peptide has been implicated in the pathogenesis of EDN, but the mechanisms of these effects have not been fully characterized. The aims of this study were therefore to explore the effects of diabetes on neurovascular dysfunction and indexes of nerve oxidative stress in type 1 bio-breeding Worcester (BB/Wor) rats and type 2 BB Zucker-derived (ZDR)/Wor rats and to determine the effects of C-peptide replacement in the former. Motor and sensory nerve conduction velocities (NCVs), hindlimb thermal thresholds, endoneurial blood flow, and indicators of oxidative stress were evaluated in nondiabetic control rats, BB/Wor rats, BB/Wor rats with rat II C-peptide replacement (75 nmol C-peptide.kg body wt(-1).day(-1)) for 2 mo, and diabetes duration-matched BBZDR/Wor rats. Endoneurial perfusion was decreased and oxidative stress increased in type 1 BB/Wor rats. C-peptide prevented NCV and neurovascular deficits and attenuated thermal hyperalgesia. Inhibition of nitric oxide (NO) synthase, but not cyclooxygenase, reversed the C-peptide-mediated effects on NCV and nerve blood flow. Indexes of oxidative stress were unaffected by C-peptide. In type 2 BBZDR/Wor rats, neurovascular deficits and increased oxidative stress were unaccompanied by sensory NCV slowing or hyperalgesia. Therefore, nerve oxidative stress is increased and endoneurial perfusion decreased in type 1 BB/Wor and type 2 BBZDR/Wor rats. NO and neurovascular mechanisms, but not oxidative stress, appear to contribute to the effects of C-peptide in type 1 EDN. Sensory nerve deficits are not an inevitable consequence of increased oxidative stress and decreased nerve perfusion in a type 2 diabetic rodent model.


Is There Any Effect of Subjective Tinnitus on Vestibular Evoked Myogenic Potentials.
Idiopathic subjective tinnitus has a complex pathophysiology in which not only cochlear and central classical auditory pathways but also nonclassical auditory pathways of different parts of the brain are involved. Vestibuloocular and vestibulocollic pathways are the central projections of utricle and saccule used in the vestibular evoked myogenic potential (VEMP) test. Aim of this study was to investigate the effects of idiopathic subjective tinnitus on vestibuloocular and vestibulocollic pathways via VEMP. We prospectively analyzed 30 unilateral idiopathic subjective tinnitus patient's cervical, ocular VEMP tests, tinnitus handicap index scores, symptom duration and compared with contralateral ear and 35 healthy volunteers. The latencies and amplitudes of P1 and N1 waves were recorded and pathologic wave criteria was calculated according to healthy volunteer's data. In cervical VEMP there were significant longer latencies of P1 and N1 waves with respect to contralateral ear and control group. In ocular VEMP test, N1 and P1 latencies and amplitudes were not significantly different. The percentages of pathologic wave of the tinnitus side were not significantly higher in both cervical VEMP and ocular VEMP tests with respect to contralateral side. Tinnitus handicap index scores and symptom duration had no relationship with latency and amplitude of VEMP tests. Although cervical VEMP P1 and N1 latencies were significantly longer, subjective tinnitus did not result in pathological alterations in the VEMP test. Presence of subjective tinnitus is not an influencing factor in the VEMP interpretation.


TEAM to Defeat COVID-19: A Management Strategy Plan to Address Return to Play in Sports Medicine.
A new viral disease named COVID-19 spread rapidly into a pandemic in early 2020. Most countries have active community transmission and imposed measures such as social distancing and travel restrictions to mitigate its effect. Many sporting events all over the globe were canceled or postponed. In this article, we briefly discuss some important topics regarding the COVID-19 pandemic and propose a strategy to manage return to play in professional athletes. We searched the PubMed and Google Scholar databases to identify articles published through May 12, 2020, using the following keywords: "coronavirus," "COVID-19," "SARS-CoV-2," "athlete," and "return to play." Although athletes have a lower risk of severe disease, preventive measures are still very important for minimizing time away from training, avoiding potential complications, slowing the pandemic spread, and ultimately protecting the health of those with a higher risk of complications and death. Athletes may present with mild disease, but complications such as pulmonary fibrosis and myocardial injuries have to be considered. Although still controversial, athletes should be evaluated before return to play and monitored accordingly afterward via proper clinical assessments and testing.


Listeria monocytogenes-associated respiratory infections: a study of 38 consecutive cases.
Listeria monocytogenes (Lm) is a foodborne human pathogen responsible for severe infections, including septicaemia, neurolisteriosis, and maternal-foetal and focal infections. Little is known about Lm-associated respiratory tract or lung infections. We conducted a retrospective study of culture-proven cases of Lm pleural infections and pneumonia reported to the French National Reference Centre for Listeria from January 1993 to August 2016. Thirty-eight consecutive patients with pleural infection (n = 32), pneumonia (n = 5), or both (n = 1) were studied; 71% of these were men. Median age was 72 (range 29-90). Two patients presented with concomitant neurolisteriosis. All patients but one reported at least one immunosuppressive condition (97%), with a median number of 2 (range 0-5), including 29% (8/28) with current exposure to immunosuppressive therapy and 50% (17/34) with ongoing neoplasia; 75% (21/28) reported previous pleural or pulmonary disease. Antibiotic therapy mostly consisted in amoxicillin (72%) associated with aminoglycoside in 32%. Chest-tube drainage was performed in 7/19 patients with empyema (37%); 25% of the patients (7/30) required intensive care management. In-hospital mortality reached 35% and occurred after a median time interval of 4 days (range 1-33 days). Three patients had recurrence of empyema (time interval of 1 week to 4 months after treatment completion). Altogether, only 13/31 patients (42%) diagnosed with Lm respiratory infection experienced an uneventful outcome at 2-year follow-up. Lm is a rare but severe cause of pneumonia and pleural infection in older immunocompromised patients, requiring prompt diagnosis and adequate management and follow-up.


Stochastic shielding and edge importance for Markov chains with timescale separation.
Nerve cells produce electrical impulses ("spikes") through the coordinated opening and closing of ion channels. Markov processes with voltage-dependent transition rates capture the stochasticity of spike generation at the cost of complex, time-consuming simulations. Schmandt and Galán introduced a novel method, based on the stochastic shielding approximation, as a fast, accurate method for generating approximate sample paths with excellent first and second moment agreement to exact stochastic simulations. We previously analyzed the mathematical basis for the method's remarkable accuracy, and showed that for models with a Gaussian noise approximation, the stationary variance of the occupancy at each vertex in the ion channel state graph could be written as a sum of distinct contributions from each edge in the graph. We extend this analysis to arbitrary discrete population models with first-order kinetics. The resulting decomposition allows us to rank the "importance" of each edge's contribution to the variance of the current under stationary conditions. In most cases, transitions between open (conducting) and closed (non-conducting) states make the greatest contributions to the variance, but there are exceptions. In a 5-state model of the nicotinic acetylcholine receptor, at low agonist concentration, a pair of "hidden" transitions (between two closed states) makes a greater contribution to the variance than any of the open-closed transitions. We exhaustively investigate this "edge importance reversal" phenomenon in simplified 3-state models, and obtain an exact formula for the contribution of each edge to the variance of the open state. Two conditions contribute to reversals: the opening rate should be faster than all other rates in the system, and the closed state leading to the opening rate should be sparsely occupied. When edge importance reversal occurs, current fluctuations are dominated by a slow noise component arising from the hidden transitions.


Caffeine and seizures: A systematic review and quantitative analysis.
Caffeine is the most commonly used central nervous system (CNS) stimulant. The relationship between caffeine, seizures, epilepsy, and antiepileptic drugs (AEDs) is complex and not fully understood. Case reports suggest that caffeine triggers seizures in susceptible people. Our systematic review reports on the relationship between caffeine, seizures, and drugs in animal and human studies. Quantitative analyses were also done on animal studies regarding the effects of caffeine on AEDs. PubMed was searched for studies assessing the effects of caffeine on seizure susceptibility, epilepsy, and drug interactions in people and in animal models. To quantify the interaction between AEDs and caffeine, the data of six animal studies were pooled and analyzed using a general linear model univariate analysis or One-way Analysis of Variance (ANOVA). In total, 442 items were identified from which we included 105 studies. Caffeine can increase seizure susceptibility and protect from seizures, depending on the dose, administration type (chronic or acute), and the developmental stage at which caffeine exposure started. In animal studies, caffeine decreased the antiepileptic potency of some drugs; this effect was strongest in topiramate. Preclinical studies suggest that caffeine increases seizure susceptibility. In some cases, chronic use of caffeine may protect against seizures. Caffeine lowers the efficacy of several drugs, especially topiramate. It is unclear how these findings in models can be translated to the clinical condition. Until clinical studies suggest otherwise, caffeine intake should be considered as a factor in achieving and maintaining seizure control in epilepsy.


Saccharomyces cerevisiae produces a yeast substance that exhibits estrogenic activity in mammalian systems.
Partially purified lipid extracts of Saccharomyces cerevisiae contain a substance that displaces tritiated estradiol from rat uterine cytosol estrogen receptors. The yeast product induces estrogenic bioresponses in mammalian systems as measured by induction of progesterone receptors in cultured MCF-7 human breast cancer cells and by a uterotrophic response and progesterone receptor induction after administration to ovariectomized mice. The findings raise the possibility that bakers' yeast may be a source of environmental estrogens.


Buying Time: Using OMM to Potentially Reduce the Demand for Mechanical Ventilation in Patients With COVID-19.
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) pandemic is causing an increased need for mechanical ventilation for a significant percentage of people who present to the hospital for treatment. This increase in demand could surpass the supply of ventilators and lead to an increase in mortality due to a lack of ventilator vacancies. There is significant evidence that osteopathic manipulative medicine (OMM) can alleviate pulmonary symptoms and aid in quicker recoveries from various respiratory ailments. OMM has the potential to play a significant role in helping reduce a patient's need for mechanical ventilation by delaying the onset of acute respiratory distress syndrome stemming from SARS-Cov2 infections.


Composite grafting for distal digital amputation with respect to injury type and amputation level.
This study evaluated the composite graft survival rate in distal digital amputations with respect to injury type and amputation level. Twenty-seven patients with complete fingertip amputations (32 digits) distal to the distal interphalangeal joint who were treated by composite grafting from January 2010 to February 2012 were enrolled. Injury type was classified as clean-cut, blunt-cut, or crush-avulsion. Amputation level was classified according to Ishikawa's classification: subzones I-IV. Graft survival was categorised as complete, partial, or no survival. The graft was more likely to exhibit complete survival in clean-cut injuries (50%) than in blunt-cut (10%) or crush-avulsion injuries (12.5%). However, when complete and partial survival were combined, there was no significant difference among injury types (clean-cut = 83.3%, blunt-cut = 70.0%; crush-avulsion = 68.8%). Composite grafting in sub-zone I provided good results (complete survival = 50%; partial survival = 50.0%; no survival = 0%). When complete and partial survival were combined, there was no significant difference with respect to amputation level except sub-zone I (II = 70.6%; III = 66.7%; IV = 60%). In sub-zone II, clean-cut injuries exhibited better graft survival than blunt-cut or crush-avulsion injuries. In sub-zones III and IV, no complete graft survival was observed. In conclusion, all types of injuries in sub-zone I and clean-cut injuries in sub-zone II are candidates for composite grafting. Blunt-cut and crush-avulsion injuries in sub-zone II are marginal candidates for composite grafting. Any type of injury in sub-zone III or IV is contraindicated for composite grafting and should be treated by microanastomosis.


Plasma linoleic acid partially mediates the association of bipolar disorder on self-reported mental health scales.
We have shown that bipolar individuals have reduced quality diets, including lower intake of polyunsaturated fatty acids (PUFA). We have also reported reduced plasma levels of the n-6 PUFA, linoleic acid (LA), and the n-3 PUFA, eicosapentaenoic acid (EPA) in bipolar subjects. In the current analysis we hypothesized that LA and EPA plasma levels would mediate lower self-reported mental health and life functioning scores in bipolar subjects. In a cross-sectional study, we collected a 7-day diet record in bipolar (n = 56) and control subjects (n = 46) followed by a fasted blood draw. We used structured equation modeling path analysis to test for mediating effects of dietary intake and plasma levels of LA and EPA on self-reported mental health questionnaire scores, including the Life Functioning Questionnaire (LFQ), the Patient Health Questionnaire (PHQ9), and the Short Form Health Survey (SF12), extracting the mental health component summary score (SF12-MH). We adjusted for age, gender, psychiatric medication use, body mass index (BMI), and total caloric intake as covariates with bipolar disorder as the primary predictor. We found a significant path association from bipolar disorder to lower plasma LA levels (p = 0.03) and significant paths from plasma LA to PHQ9 (p = 0.05), LFQ (p = 0.01) and SF12-MH (p = 0.05) scores, such that lower plasma LA predicted worse outcomes. We found no significant paths from plasma EPA levels to any of the outcome measures. These findings suggest that plasma LA levels partially mediate the effect of bipolar disorder on self-reported measures of mental health and life functioning.


Tumor necrosis factor (TNF) interferes with insulin signaling through the p55 TNF receptor death domain.
Tumor necrosis factor (TNF) contributes to insulin resistance by binding to the 55kDa TNF receptor (TNF-R55), resulting in serine phosphorylation of proteins such as insulin receptor (IR) substrate (IRS)-1, followed by reduced tyrosine phosphorylation of IRS-1 through the IR and, thereby, diminished IR signal transduction. Through independent receptor domains, TNF-R55 activates a neutral (N-SMase) and an acid sphingomyelinase (A-SMase), that both generate the sphingolipid ceramide. Multiple candidate kinases have been identified that serine-phosphorylate IRS-1 in response to TNF or ceramide. However, due to the fact that the receptor domain of TNF-R55 mediating inhibition of the IR has not been mapped, it is currently unknown whether TNF exerts these effects with participation of N-SMase or A-SMase. Here, we identify the death domain of TNF-R55 as responsible for the inhibitory effects of TNF on tyrosine phosphorylation of IRS-1, implicating ceramide generated by A-SMase as a downstream mediator of inhibition of IR signaling.


[Anopheles (Díptera: Culicidae) vectors of malaria in Puerto Carreño municipality, Vichada, Colombia].
The study of the biological aspects of Anopheles spp., strengthens the entomological surveillance. To determine biological aspects and behavior of adult Anopheles mosquitoes in the urban area of Puerto Carreño municipality, Vichada, Colombia. Wild anophelines were collected landing on humans both indoors and outdoors between 18:00h and 06:00h for 50 min/h during two consecutive nights/month for eight months in the urban area of Puerto Carreño. The biting rate activity, the natural infection by Plasmodium falciparum and P. vivax VK247 and VK210 using ELISA, and the annual entomological inoculation rate were determined for each species. The members of the Albitarsis complex were determined by amplificacion of the white gene by polymerase chain reaction. In order of abundance the species found were An. darlingi (n=1,166), An. marajoara sensu stricto (n=152), An. braziliensis (n=59), An. albitarsis F (n=25), An. albitarsis sensu lato (n=16), An. argyritarsis (n=3) and An. oswaldoi sensu lato (n=2). An. darlingi showed two activity peaks between 21:00 to 22:00 and 05:00 to 06:00 hours outdoors and between 21:00 to 22:00 and 04:00 to 05:00 indoors. Natural infection of this species was found with P. vivax VK210 and its annual entomological inoculation rate was 2. Natural infection of An marajoara sensu stricto with P. falciparum was found, with an annual entomological inoculation rate of 5 and a peak biting activity between 18:00 to 19:00 hrs both indoors and outdoors. Transmission of malaria in the urban area of Puerto Carreño, Vichada, can occur by An. darlingi and An. marajoara s.s.


Recommendations for the establishment of primary stroke centers. Brain Attack Coalition.
To develop recommendations for the establishment and operation of primary stroke centers as an approach to improve the medical care of patients with stroke. Members of the Brain Attack Coalition (BAC), a multidisciplinary group of representatives from major professional organizations involved with delivering stroke care. Supplemental input was obtained from other experts involved in acute stroke care. A review of literature published from 1966 to March 2000 was performed using MEDLINE. More than 600 English-language articles that had evidence from randomized clinical trials, meta-analyses, care guidelines, or other appropriate methods supporting specific care recommendations for patients with acute stroke that could be incorporated into a stroke center model were selected. Articles were reviewed initially by 1 author (M.J.A.). Members of the BAC reviewed each recommendation in the context of current practice parameters, with special attention to improving the delivery of care to patients with acute stroke, cost-effectiveness, and logistical issues related to the establishment of primary stroke centers. Consensus was reached among all BAC participants before an element was added to the list of recommendations. Randomized clinical trials and observational studies suggest that several elements of a stroke center would improve patient care and outcomes. Key elements of primary stroke centers include acute stroke teams, stroke units, written care protocols, and an integrated emergency response system. Important support services include availability and interpretation of computed tomography scans 24 hours everyday and rapid laboratory testing. Administrative support, strong leadership, and continuing education are also important elements for stroke centers. Adoption of these recommendations may increase the use of appropriate diagnostic and therapeutic modalities and reduce peristroke complications. The establishment of primary stroke centers has the potential to improve the care of patients with stroke. JAMA. 2000.


Polyamide nucleic acid-DNA chimera lacking the phosphate backbone are novel primers for polymerase reaction catalyzed by DNA polymerases.
A peptide nucleic acid (PNA) oligomer, an analogue of DNA, was examined for its ability to function as a primer or a template to support DNA synthesis catalyzed by DNA polymerases. The analogue, (PNA)19-TpG-OH, comprised of 19 bases in the form of PNA followed by a dinucleotide (TpG-OH) with a single phosphate and a free 3'OH terminus, was recognized as a bona fide primer by 2 reverse transcriptases and also by the Klenow fragment of E. coli DNA polymerase I. The 21-mer PNA chimera is extended on both RNA and DNA templates by all three polymerases. The specificity of binding of the PNA chimeric primer/DNA template at the template-primer binding site of the enzyme was shown by its photo-cross-linking ability to the enzyme which could be effectively competed out by another TP but not by template or primer alone. Furthermore, the chimeric TP-enzyme covalent complex was found to be catalytically active as judged by its ability to incorporate one nucleotide onto the 3'OH terminus of the immobilized primer. PNA sequences were also recognized as template when annealed with a DNA primer. These observations are in variance with the conventional suggestion that the phosphate backbone in the duplex region is essential for recognition and binding by DNA polymerases. The efficient extension of (PNA)19-TpG-OH suggests that the diameter of the duplex region of template primer rather than the phosphate backbone may be sufficient for recognition by DNA polymerases.


Studies on renal juxtaglomerular cells. I. Variations produced by sodium chloride and desoxycorticosterone acetate.
Accumulation of granules in the juxtaglomerular cells occurred in rats which were maintained for 5 to 6 weeks on a diet low in sodium, chloride. Cytological evidence suggests that this was probably a storage phase of secretion following a decrease in the rate of liberation of the granules. Administration of DCA (desoxycorticosterone acetate) to salt-deficient rats did not alter this appearance of the juxtaglomerular cells. Two per cent sodium chloride taken in the drinking water consumed for 4 weeks by similar animals caused degranulation of the juxtaglomerular cells. This effect was enhanced by DCA. DCA administered to animals on a normal salt intake produced a lesser degree of degranulation. Cytological changes in degranulated cells suggested that these represent a stage of hyperactivity in the secretory cycle produced by an increase in the rate of liberation of granules. A hypothesis is suggested that the juxtaglomerular cells are involved in the hormonal regulation of sodium metabolism and/or blood pressure.


Adopting leisure-time physical activity after diagnosis of a vascular condition.
A better understanding of factors associated with adopting leisure-time physical activity among people with chronic vascular conditions can help policy-makers and health care professionals develop strategies to promote secondary prevention among older Canadians. Cross-sectional data from the 1994/1995 National Population Health Survey (NPHS), household component, and the 2007/2008 Canadian Community Health Survey were used to estimate the prevalence of inactivity. Longitudinal data from eight cycles (1994/1995 through 2008/2009) of the NPHS, household component, were used to examine the adoption of leisure-time physical activity, intentions to change health risk behaviours, and barriers to change. Over half (54%) of the population aged 40 or older were inactive during their leisure time in 2007/2008. A new vascular diagnosis was not associated with initiating leisure-time physical activity. Among the newly diagnosed, those with no disability or a mild disability had higher odds of undertaking leisure-time physical activity. The majority of Canadians in mid- to late life are inactive. They tend to remain so when diagnosed with a vascular condition.


Fundamentals of soft thermofluidic system design.
The soft composition of many natural thermofluidic systems allows them to effectively move heat and control its transfer rate by dynamically changing shape (e.g. dilation or constriction of capillaries near our skin). So far, making analogous deformable "soft thermofluidic systems" has been limited by the low thermal conductivity of materials with suitable mechanical properties. By remaining soft and stretchable despite the addition of filler, elastomer composites with thermal conductivity enhanced by liquid-metal micro-droplets provide an ideal material for this application. In this work, we use these materials to develop an elementary thermofluidic system consisting of a soft, heat generating pipe that is internally cooled with flow of water and explore its thermal behavior as it undergoes large shape change. The transient device shape change invalidates many conventional assumptions employed in thermal design making analysis of this devices' operation a non-trivial undertaking. To this end, using time scale analysis we demonstrate when the conventional assumptions break down and highlight conditions under which the quasi-static assumption is applicable. In this gradual shape modulation regime the actuated devices' thermal behavior at a given stretch approaches that of a static device with equivalent geometry. We validate this time scale analysis by experimentally characterizing thermo-fluidic behavior of our soft system as it undergoes axial periodic extension-retraction at varying frequencies during operation. By doing so we explore multiple shape modulation regimes and provide a theoretical foundation to be used in the design of soft thermofluidic systems undergoing transient deformation.


Generation and control of sound bullets with a nonlinear acoustic lens.
Acoustic lenses are employed in a variety of applications, from biomedical imaging and surgery to defense systems and damage detection in materials. Focused acoustic signals, for example, enable ultrasonic transducers to image the interior of the human body. Currently however the performance of acoustic devices is limited by their linear operational envelope, which implies relatively inaccurate focusing and low focal power. Here we show a dramatic focusing effect and the generation of compact acoustic pulses (sound bullets) in solid and fluid media, with energies orders of magnitude greater than previously achievable. This focusing is made possible by a tunable, nonlinear acoustic lens, which consists of ordered arrays of granular chains. The amplitude, size, and location of the sound bullets can be controlled by varying the static precompression of the chains. Theory and numerical simulations demonstrate the focusing effect, and photoelasticity experiments corroborate it. Our nonlinear lens permits a qualitatively new way of generating high-energy acoustic pulses, which may improve imaging capabilities through increased accuracy and signal-to-noise ratios and may lead to more effective nonintrusive scalpels, for example, for cancer treatment.


Spontaneous seroclearance of hepatitis B seromarkers and subsequent risk of hepatocellular carcinoma.
The associations between long-term risk of hepatocellular carcinoma (HCC) and spontaneous seroclearance of HBV e antigen (HBeAg), HBV DNA and HBV surface antigen (HBsAg) have never been examined by a prospective study using serially measured seromarkers. This study aimed to assess the importance of spontaneous HBeAg, HBV DNA and HBsAg seroclearance in the prediction of HCC risk. This study included 2946 HBsAg seropositive individuals who were seronegative for antibodies against HCV and free of liver cirrhosis. Serial serum samples collected at study entry and follow-up health examinations were tested for HBeAg, HBV DNA and HBsAg. Cox proportional hazards models were used to calculate the HRs of developing HCC after seroclearance of HBV markers. The HR (95% CI) of developing HCC after seroclearance of HBeAg, HBV DNA and HBsAg during follow-up was 0.63 (0.38 to 1.05), 0.24 (0.11 to 0.57) and 0.18 (0.09 to 0.38), respectively, after adjustment for age, gender and serum level of alanine aminotransferase at study entry. High HBV DNA levels at the seroclearance of HBeAg (mean±SD, 4.35±1.64 log10 IU/mL) may explain the non-significant association between HBeAg seroclearance and HCC risk. Among HBeAg seronegative participants with detectable serum HBV DNA at study entry, the lifetime (30-75-years-old) cumulative incidence of HCC was 4.0%, 6.6% and 14.2%, respectively, for those with seroclearance of both HBV DNA and HBsAg, seroclearance of HBV DNA only, and seroclearance of neither. Spontaneous seroclearance of HBV DNA and HBsAg are important predictors of reduced HCC risk.


Idiopathic eruptive macular pigmentation with papillomatosis: Report of nine cases.
Nine patients, seven males and two females aged 6-14 years, presented with extensive, asymptomatic, brown-black macules and mildly elevated, pigmented lesions of a few months' duration. The sites affected were the face, trunk and proximal extremities. The skin lesions were discrete and individual lesions were less than 2 cm in size. The clinical diagnoses rendered by the referring physicians were lichen planus pigmentosus, urticaria pigmentosa, erythema dyschromicum perstans and postinflammatory hyperpigmentation. Histology in all nine cases showed papillomatosis of the dermis with prominent pigmentation of the basal layer (pigmented papillomatosis) without any significant dermal inflammation. Two cases had spores of Pityrosporum ovale in the thickened horny layer, one of which also had, in addition, bacterial colonies in the stratum corneum. The pigmentation resolved on its own over several months. This presentation is similar to the previously described idiopathic eruptive macular pigmentation with the additional histological finding of papillomatosis that is being described for the first time and may be nosologically related to acanthosis nigricans and confluent and reticulate papillomatosis.


K63-Linked Ubiquitination Targets Toxoplasma gondii for Endo-lysosomal Destruction in IFNγ-Stimulated Human Cells.
Toxoplasma gondii is the most common protozoan parasitic infection in man. Gamma interferon (IFNγ) activates haematopoietic and non-haematopoietic cells to kill the parasite and mediate host resistance. IFNγ-driven host resistance pathways and parasitic virulence factors are well described in mice, but a detailed understanding of pathways that kill Toxoplasma in human cells is lacking. Here we show, that contrary to the widely held belief that the Toxoplasma vacuole is non-fusogenic, in an immune-stimulated environment, the vacuole of type II Toxoplasma in human cells is able to fuse with the host endo-lysosomal machinery leading to parasite death by acidification. Similar to murine cells, we find that type II, but not type I Toxoplasma vacuoles are targeted by K63-linked ubiquitin in an IFNγ-dependent manner in non-haematopoetic primary-like human endothelial cells. Host defence proteins p62 and NDP52 are subsequently recruited to the type II vacuole in distinct, overlapping microdomains with a loss of IFNγ-dependent restriction in p62 knocked down cells. Autophagy proteins Atg16L1, GABARAP and LC3B are recruited to <10% of parasite vacuoles and show no parasite strain preference, which is consistent with inhibition and enhancement of autophagy showing no effect on parasite replication. We demonstrate that this differs from HeLa human epithelial cells, where type II Toxoplasma are restricted by non-canonical autophagy leading to growth stunting that is independent of lysosomal acidification. In contrast to mouse cells, human vacuoles do not break. In HUVEC, the ubiquitinated vacuoles are targeted for destruction in acidified LAMP1-positive endo-lysosomal compartments. Consequently, parasite death can be prevented by inhibiting host ubiquitination and endosomal acidification. Thus, K63-linked ubiquitin recognition leading to vacuolar endo-lysosomal fusion and acidification is an important, novel virulence-driven Toxoplasma human host defence pathway.


Cancer in first-degree relatives and risk of glioma in adults.
Relatively few studies have examined glioma risk in relation to history of cancer in first-degree relatives. We sought to describe such risks in a large hospital-based case-control study. Histologically confirmed incident adult glioma cases (n = 489) were identified at three regional referral hospitals between June 1994 and August 1998. Controls (n = 799) admitted to the same hospitals for nonmalignant conditions were frequency-matched on age, sex, race/ethnicity, hospital, and proximity of residence to hospital. Participants received a personal interview, including questions regarding cancer in family members. Odds ratios (ORs) were calculated to estimate the risk of glioma associated with a history of cancer in a first-degree relative using conditional logistic regression and compared with standardized incidence ratios among relatives of cases versus relatives of controls. Among participants reporting a family history of a brain cancer or a brain tumor, risk of glioma was 1.6 [95% confidence interval (CI), 0.5-5.3; n = 5] and 3.0 (95% CI, 0.9-10.8; n = 7), respectively, in comparison with those without such family histories. Participants who had a family history of stomach (OR, 2.2; 95% CI, 1.0-4.6), colon (OR, 1.4; 95% CI, 0.9-2.2), or prostate cancer (OR, 2.1; 95% CI, 1.1-3.8) or Hodgkin disease (OR, 2.4; 95% CI, 0.9-6.3) had an increased glioma risk. OR estimates were similar to the ratios of standardized incidence ratios for cancer in relatives of cases versus controls. Shared environmental or genetic factors in families may influence glioma risk. Our findings suggest that individuals with a family history of specific cancers other than glioma may have an increased glioma risk.


AidP, a novel N-Acyl homoserine lactonase gene from Antarctic Planococcus sp.
Planococcus is a Gram-positive halotolerant bacterial genus in the phylum Firmicutes, commonly found in various habitats in Antarctica. Quorum quenching (QQ) is the disruption of bacterial cell-to-cell communication (known as quorum sensing), which has previously been described in mesophilic bacteria. This study demonstrated the QQ activity of a psychrotolerant strain, Planococcus versutus strain L10.15T, isolated from a soil sample obtained near an elephant seal wallow in Antarctica. Whole genome analysis of this bacterial strain revealed the presence of an N-acyl homoserine lactonase, an enzyme that hydrolyzes the ester bond of the homoserine lactone of N-acyl homoserine lactone (AHLs). Heterologous gene expression in E. coli confirmed its functions for hydrolysis of AHLs, and the gene was designated as aidP (autoinducer degrading gene from Planococcus sp.). The low temperature activity of this enzyme suggested that it is a novel and uncharacterized class of AHL lactonase. This study is the first report on QQ activity of bacteria isolated from the polar regions.


Effect of race on the presentation and management of patients with acute chest pain.
To compare racial differences in clinical presentation, natural history, and access to medical care and procedures among emergency-department patients with acute chest pain. Prospective follow-up study of consecutive patients coming to the emergency department because of acute chest pain. Two university medical centers. A total of 3031 patients who were 30 years or older and who came to the emergency department with acute chest pain from 1984 to 1986. African-Americans tended to have slightly, but not always significantly, lower rates of acute myocardial infarction, acute ischemic heart disease, and major complications, after adjusting for presenting symptoms and signs; the adjusted odds ratios for African-Americans were as follows: 0.77 (95% CI, 0.54 to 1.1) for acute myocardial infarction, 0.75 (CI, 0.59 to 0.95) for ischemic heart disease, and 0.79 (CI, 0.45 to 1.4) for death or major complications. Clinical factors classically associated with acute myocardial infarction were equally predictive in African-Americans and whites. After adjustments were made for multiple clinical factors, a lower proportion of African-Americans were admitted to the hospital (odds ratio, 0.69; CI, 0.56 to 0.84), and, once admitted, were somewhat less likely to be triaged to the coronary care unit (odds ratio, 0.81; CI, 0.65 to 1.0). In adjusted analyses, African-Americans were as likely to undergo cardiac catheterization as whites (odds ratio, 0.86; CI, 0.64 to 1.2) but were less likely to undergo coronary artery bypass procedures once severity of coronary disease was included in the analysis (odds ratio, 0.24; CI, 0.08 to 0.71). African-Americans and whites had a similar presentation and natural history of acute myocardial infarction and, after adjusting for probability of clinical events, similar access to most medical care and cardiac procedures. However, the rate of coronary artery bypass procedures was much lower among African-Americans than among whites. Reasons for this difference should be studied.


Delayed reaction to cow's milk proteins: a case study.
Allergy to cow's milk proteins has been defined as any adverse reaction mediated by immunological mechanisms to one or several of these proteins. The diagnosis can be made based on clinical manifestations supported by immune activation of in vitro parameters. Reactions to cow's milk have been classified according on their onset as immediate (< 45 min) or delayed-type (from 2 h to days). We describe a patient with late respiratory manifestations after milk intake, probably due to more than one immunological mechanism. He was an 18-year-old male who since infancy had presented serous rhinorrea, sneezing, nasal blockade, oropharyngeal pruritus and occasional dyspnea 12 to 48 h after ingestion of milk and its derivates. We performed skin prick and intradermal tests with whole milk and fractions. Patch tests were also carried out with whole milk purchased at a supermarket and with the extracts described, in their original form and vehiculized in vaseline. Total and serum specific IgE and IgG4 to milk fractions, histamine release test (HRT) to milk fractions, and precipitating antibodies by contraimmunoelectrophoresis against milk fractions were also measured. As a control we repeated this test in a patient with IgE-mediated manifestations to milk proteins and in two healthy controls. We performed a single-blind placebo controlled challenge with whole milk. Skin prick and intradermal tests were negative. Patch test (48 h) was positive for whole milk and whole milk vehiculized in vaseline, and for alpha-lactalbumin. Total IgE was 559 kU/l; serum-specific IgE was negative; IgG4 was positive (9.48% for alpha-lactalbumin; 7.41% for beta-lactoglobulin and 9.85% for casein). HRT was positive for casein (34%). We found precipitating antibodies to the three milk fractions in our patient and in the atopic control. In the challenge test, 10 h after milk intake the patient presented serous rhinorrea, sneezing and nasal blockade. IgG4 was involved as a blocking or anaphylactic antibody and as an immunological epiphenomenon reflecting a permanent antigenic stimulus. We find this last explanation to be the most coherent in this case.


Two promoter rearrangements in a drug efflux transporter gene are responsible for the appearance and spread of multidrug resistance phenotype MDR2 in Botrytis cinerea isolates in French and German vineyards.
In French and German vineyards, Botrytis cinerea isolates with multiple fungicide resistance phenotypes have been observed with increasing frequencies. Multidrug resistance (MDR) results from mutations that lead to constitutive overexpression of genes encoding drug efflux transporters. In MDR2 and MDR3 strains, overexpression of the major facilitator superfamily transporter gene mfsM2 has been found to result from a rearrangement in the mfsM2 promoter (type A), caused by insertion of a retroelement (RE)-derived sequence. Here, we report the discovery of another, similar RE-induced rearrangement of the mfsM2 promoter (type B) in a subpopulation of French MDR2 isolates. MDR2 isolates harboring either type A or type B mutations in mfsM2 show the same resistance phenotypes and similar levels of mfsM2 overexpression. RE sequences similar to those in mfsM2 were found in low copy numbers in other but not all B. cinerea strains analyzed, including non-MDR2 strains. Population genetic analyses support the hypothesis that the two rearrangement mutations have only occurred once, and are responsible for the appearance and subsequent spread of all known MDR2 and MDR3 strains in French and German wine-growing regions.


Detection of HP10 antigen in serum for diagnosis and follow-up of subarachnoidal and intraventricular human neurocysticercosis.
Neurocysticercosis (NC), a parasitic disease caused by Taenia solium, may be either asymptomatic or show a mild to severe clinical picture with intracranial hypertension. The most severe form of the disease is caused when viable cysticerci are localised in the ventricles or in subarachnoidal cisterns at the base of the skull. Detection of the secreted metacestode antigen HP10 in cerebrospinal fluid is a sensitive and specific method for the diagnosis of these severe NC cases. To evaluate the validity of HP10 antigen detection ELISA when applied to serum, using paired serum and cerebrospinal fluid samples from 116 radiologically and clinically characterised NC patients. The HP10 antigen assay exhibited a similarly high sensitivity in identifying severe NC cases from sera (84.8%) and CSF (91.3%). In contrast, HP10 antigen was rarely detected in asymptomatic or mild NC cases (3 of 57). Importantly, the HP10 antigen assay applied to serum showed high specificity (94%) when used in 126 serum samples of non-NC subjects from an endemic community with a confirmed coproparasitological diagnosis of intestinal parasitic infections. Finally, the HP10 assay also proved to be of value in the follow-up of treated patients. This study confirms that detection of the metacestode HP10 antigen in serum is a useful tool for diagnosis and follow-up of patients with severe forms of NC treated with cysticidal drugs.


An MRI study of spatial probability brain map differences between first-episode schizophrenia and normal controls.
We created a spatial probability atlas of schizophrenia to provide information about the neuroanatomic variability of brain regions of patients with the disorder. Probability maps of 16 regions of interest (ROIs) were constructed by taking manually parcellated ROIs from subjects' magnetic resonance images (MRIs) and linearly transforming them into Talairach space using the Montreal Neurological Institute (MNI) template. ROIs included temporal, parietal, and prefrontal cortex subregions, with a principal focus on temporal lobe structures. Subject Ns ranged from 11 to 28 for the different ROIs. Our global measure of the spatial distribution of the transformed ROI was the sum of voxels with 50% overlap among subjects. The superior temporal gyrus (STG) and fusiform gyrus (FG) had lower values for schizophrenic subjects than for normal controls, suggestive of greater spatial variability for these ROIs in schizophrenic subjects. For the computation of statistical significance of group differences in portions of the ROI, we used voxel-wise comparisons and Fisher's exact test. First-episode schizophrenic patients compared with controls showed lower probability (P < 0.05) at dorso-posterior areas of planum temporale and Heschl's gyrus, lateral and anterior regions in the left hippocampus (HIPP), and dorsolateral regions of fusiform gyrus. Importantly, most ROIs of schizophrenic subjects showed a significantly lower spatial overlap than controls, even after nonlinear spatial normalization, suggesting a greater heterogeneity in the spatial distribution of ROIs. There is consequently a need for caution in neuroimaging studies where data from schizophrenic subjects are normalized to a particular stereotaxic coordinate system based on healthy controls. Apparent group differences in activation may simply reflect a greater heterogeneity of spatial distribution in schizophrenia.


Unbalanced segregation of a paternal t(9;11)(p24.3;p15.4) translocation causing familial Beckwith-Wiedemann syndrome: a case report.
The vast majority of cases with Beckwith-Wiedemann syndrome (BWS) are caused by a molecular defect in the imprinted chromosome region 11p15.5. The underlying mechanisms include epimutations, uniparental disomy, copy number variations, and structural rearrangements. In addition, maternal loss-of-function mutations in CDKN1C are found. Despite growing knowledge on BWS pathogenesis, up to 20% of patients with BWS phenotype remain without molecular diagnosis. Herein, we report an Iranian family with two females affected with BWS in different generations. Bisulfite pyrosequencing revealed hypermethylation of the H19/IGF2: intergenic differentially methylated region (IG DMR), also known as imprinting center 1 (IC1) and hypomethylation of the KCNQ1OT1: transcriptional start site (TSS) DMR (IC2). Array CGH demonstrated an 8 Mb duplication on chromosome 11p15.5p15.4 (205,827-8,150,933) and a 1 Mb deletion on chromosome 9p24.3 (209,020-1,288,114). Chromosome painting revealed that this duplication-deficiency in both patients is due to unbalanced segregation of a paternal reciprocal t(9;11)(p24.3;p15.4) translocation. This is the first report of a paternally inherited unbalanced translocation between the chromosome 9 and 11 short arms underlying familial BWS. Copy number variations involving the 11p15.5 region are detected by the consensus diagnostic algorithm. However, in complex cases which do not only affect the BWS region itself, characterization of submicroscopic chromosome rearrangements can assist to estimate the recurrence risk and possible phenotypic outcomes.


Factors affecting interindividual differences in clozapine response: a review and case report.
Clozapine is the most powerful new‐generation antipsychotic. Although this drug leads to great therapeutic benefits, two types of undesirable conditions frequently occur with its use: side effects and resistance to treatment. Therapeutic drug monitoring of clozapine would be very useful to avoid both these situations. The necessity of monitoring the therapy is the result of a wide interindividual variability in the metabolism of clozapine. In this review, we highlight all the conditions underlying this variability, analyzing them one by one. Relevant literature was identified through a search of MEDLINE and PubMed. In addition, the case of a treatment‐resistant patient with accelerated metabolism of clozapine is reported as representative of utility of therapeutic drug monitoring in terms of clozapine dose adjustment. Genetic polymorphisms of cytochrome P450 enzymes and of neurotransmitter receptors; drug interactions; interactions of clozapine with other substances such as food and drink; smoking; and nonmodifiable variables such as age, ethnicity, and gender have been examined in relation to the existing scientific literature. The laboratory techniques that clinicians could use to identify these variables and adequate therapies are also reviewed.


Forskolin suppresses delayed-rectifier K+ currents and enhances spike frequency-dependent adaptation of sympathetic neurons.
In signal transduction research natural or synthetic molecules are commonly used to target a great variety of signaling proteins. For instance, forskolin, a diterpene activator of adenylate cyclase, has been widely used in cellular preparations to increase the intracellular cAMP level. However, it has been shown that forskolin directly inhibits some cloned K+ channels, which in excitable cells set up the resting membrane potential, the shape of action potential and regulate repetitive firing. Despite the growing evidence indicating that K+ channels are blocked by forskolin, there are no studies yet assessing the impact of this mechanism of action on neuron excitability and firing patterns. In sympathetic neurons, we find that forskolin and its derivative 1,9-Dideoxyforskolin, reversibly suppress the delayed rectifier K+ current (IKV). Besides, forskolin reduced the spike afterhyperpolarization and enhanced the spike frequency-dependent adaptation. Given that IKV is mostly generated by Kv2.1 channels, HEK-293 cells were transfected with cDNA encoding for the Kv2.1 α subunit, to characterize the mechanism of forskolin action. Both drugs reversible suppressed the Kv2.1-mediated K+ currents. Forskolin inhibited Kv2.1 currents and IKV with an IC50 of ~32 μM and ~24 µM, respectively. Besides, the drug induced an apparent current inactivation and slowed-down current deactivation. We suggest that forskolin reduces the excitability of sympathetic neurons by enhancing the spike frequency-dependent adaptation, partially through a direct block of their native Kv2.1 channels.


A Combined Phenotypic and Metabolomic Approach for Elucidating the Biostimulant Action of a Plant-Derived Protein Hydrolysate on Tomato Grown Under Limited Water Availability.
Plant-derived protein hydrolysates (PHs) are an important category of biostimulants able to increase plant growth and crop yield especially under environmental stress conditions. PHs can be applied as foliar spray or soil drench. Foliar spray is generally applied to achieve a relatively short-term response, whereas soil drench is used when a long-term effect is desired. The aim of the study was to elucidate the biostimulant action of PH application method (foliar spray or substrate drench) on morpho-physiological traits and metabolic profile of tomato grown under limited water availability. An untreated control was also included. A high-throughput image-based phenotyping (HTP) approach was used to non-destructively monitor the crop response under limited water availability (40% of container capacity) in a controlled environment. Moreover, metabolic profile of leaves was determined at the end of the trial. Dry biomass of shoots at the end of the trial was significantly correlated with number of green pixels (R 2 = 0.90) and projected shoot area, respectively. Both drench and foliar treatments had a positive impact on the digital biomass compared to control while the photosynthetic performance of the plants was slightly influenced by treatments. Overall drench application under limited water availability more positively influenced biomass accumulation and metabolic profile than foliar application. Significantly higher transpiration use efficiency was observed with PH-drench applications indicating better stomatal conductance. The mass-spectrometry based metabolomic analysis allowed the identification of distinct biochemical signatures in PH-treated plants. Metabolomic changes involved a wide and organized range of biochemical processes that included, among others, phytohormones (notably a decrease in cytokinins and an accumulation of salicylates) and lipids (including membrane lipids, sterols, and terpenes). From a general perspective, treated tomato plants exhibited an improved tolerance to reactive oxygen species (ROS)-mediated oxidative imbalance. Such capability to cope with oxidative stress might have resulted from a coordinated action of signaling compounds (salicylic acid and hydroxycinnamic amides), radical scavengers such as carotenoids and prenyl quinones, as well as a reduced biosynthesis of tetrapyrrole coproporphyrins.