On the stabilization of ribose by silicate minerals.
The RNA-world theory hypothesizes that early Earth life was based on the RNA molecule. However, the notion that ribose, the sugar in RNA, is unstable still casts a serious doubt over this theory. Recently, it has been found that the silicate-mediated formose reaction facilitates the stabilization of ribose. Using accurate quantum chemical calculations, we determined the relative stability of the silicate complexes of arabinose, lyxose, ribose, and xylose with the intent to determine which would form predominantly from a formose-like reaction. Five stereoisomers were investigated for each complex. The stereoisomers of 2:1 ribose-silicate are the more stable ones, to the extent that the least stable of these is even more stable than the most stable stereoisomer of the other 2:1 sugar-silicate complexes. Thus, thermodynamically, a formose-like reaction in the presence of silicate minerals should preferentially form the silicate complex of ribose over the silicate complex of arabinose, lyxose, and xylose.


Teachers' language practices and academic outcomes of preschool children.
Early childhood programs have long been known to be beneficial to children from low-income backgrounds, but recent studies have cast doubt on their ability to substantially increase the rate of children's academic achievement. This Review examines research on the role of language in later reading, describes home and classroom factors that foster early language growth, and reviews research on preschool interventions. It argues that one reason interventions are not having as great an impact as desired is because they fail to substantially change the capacity of teachers to support children's language and associated conceptual knowledge.


Association Between Body Weight Growth and Selected Physiological Parameters in Male Japanese Quail (Coturnrix japonica).
Japanese quail is very popular research animal model. Its continued characterization for various norms is highly desirable for obtaining accurate and reliable results. This study was designed to assess various physiological parameters which are associated with body growth and development. Among various physiological parameters, blood constituents and hormones are commonly used as diagnostic tools in both physiological and pathological evaluations of humans and animals. Japanese quail hatchlings were housed in the temperature controlled brooders up to 3 weeks of age and then shifted to hanging cages in air conditioned room at ~74 F under 14L:10D lighting system and free access to feed and fresh water. Starting d8, a group of birds of uniform size and weight were selected randomly and euthanized at 4-day intervals up to d52 of age. The birds were weighed and blood sampled from the brachial vein for measuring Blood Glucose (BGL), Total Plasma Proteins (PP) and Packed Cell Volume (PCV). It was found that starting d36 all the three physiological parameters altered with approaching sexual maturity (d48-52): BGL decreased (252 vrs. 182 mg/dl, p<0.05), PCV% increased (43.6 vrs. 49.6%, p<0.05) and PP also increased (2.7 vrs. 3.2 gm/dl, p>0.05). Accordingly, BGL, PCV and PP values demonstrated significant potential to predict approaching sexual maturity in male Japanese quail.


Preschool anxiety disorders predict different patterns of amygdala-prefrontal connectivity at school-age.
In this prospective, longitudinal study of young children, we examined whether a history of preschool generalized anxiety, separation anxiety, and/or social phobia is associated with amygdala-prefrontal dysregulation at school-age. As an exploratory analysis, we investigated whether distinct anxiety disorders differ in the patterns of this amygdala-prefrontal dysregulation. Participants were children taking part in a 5-year study of early childhood brain development and anxiety disorders. Preschool symptoms of generalized anxiety, separation anxiety, and social phobia were assessed with the Preschool Age Psychiatric Assessment (PAPA) in the first wave of the study when the children were between 2 and 5 years old. The PAPA was repeated at age 6. We conducted functional MRIs when the children were 5.5 to 9.5 year old to assess neural responses to viewing of angry and fearful faces. A history of preschool social phobia predicted less school-age functional connectivity between the amygdala and the ventral prefrontal cortices to angry faces. Preschool generalized anxiety predicted less functional connectivity between the amygdala and dorsal prefrontal cortices in response to fearful faces. Finally, a history of preschool separation anxiety predicted less school-age functional connectivity between the amygdala and the ventral prefrontal cortices to angry faces and greater school-age functional connectivity between the amygdala and dorsal prefrontal cortices to angry faces. Our results suggest that there are enduring neurobiological effects associated with a history of preschool anxiety, which occur over-and-above the effect of subsequent emotional symptoms. Our results also provide preliminary evidence for the neurobiological differentiation of specific preschool anxiety disorders.


Anterior fixed prosthetic restorations and the bleached dentition: laboratory techniques.
The process of matching fixed prosthetic restorations to bleached dentition has often created a dilemma, as clinicians are not able to accurately communicate shade selection to the dental laboratory. By understanding the effect of vital bleaching on the dentition, it is possible to fabricate porcelain restorations that match the bleached natural teeth. This article presents a laboratory technique for utilizing a sequence of porcelain powders and stains to develop the internal anatomy (e.g., halos, mamelons, luminosity) of restorations that appear indistinguishable from the bleached dentition.


[Morpho-biochemical study of the liver and serum albumins during tetrachloromethane intoxication and administration of antihypoxants, antioxidants, and actoprotectors].
Organism intoxication with tetrachloromethane induces a complex of transformations in the liver and serum albumins system manifested in the liver as necroses in the central lobe, dilatation of sinus capillaries and blood stasis in them, increased content of total water, and changed magneto-relaxation properties indicating decreased structurization of water. The system of serum albumins shows decreased total concentration of albumins in the serum, decreased constant of 1-(phenylamino)-8-sulfonaphthalene probe binding, increased mean number of probe binding per albumin molecule, and increased accessibility of probe in the protein-probe complex for the quencher indicating changed type of interaction between the protein and fluorescent probe. Introduction of a antihypoxant, antioxidant, or actoprotector has a positive effect on the condition of the liver and serum albumins system: The studied indices are nearly completely normalized.


Challenges and strategies of medication adherence in Parkinson's disease: A qualitative study.
Little is known about strategies used by people with Parkinson's disease (PD) to facilitate medication adherence in the U.S. The purpose of this study was to describe challenges in adherence to medication regimens and to identify strategies used to facilitate adherence to medication regimens. A qualitative research design was used to interview sixteen community-dwelling people with PD and five caregivers. Data analysis was performed using content analysis. The majority of the participants (81.3%) reported decreased adherence to medication regimens. Seven themes emerged from the data. The main challenges of medication adherence included medication responses, cost of medications, and forgetfulness. Strategies used to facilitate adherence to medication regimens included seeking knowledge about antiparkinsonian medications, seeking advice from family and friends, use of devices, and use of reminders. These findings may be important in formulating interventions to improve adherence to medication regimens for people living with PD.


The role of bacterial infections in asthma.
In summary, bacterial organisms are clinically relevant contributors to asthma exacerbations, and have received much less attention than viruses in this process. Streptococcus pneumoniae, Hemophilus influenzae, and Moraxella catarrhalis have been linked to asthma exacerbations, particularly when sinusitis is present. Treatment therefore should be directed toward these organisms if a bacterial cause is suspected. The atypical bacteria--specifically, C. pneumoniae and M. pneumoniae--deserve special attention. Data suggest a link between these organisms and the exacerbation of asthma, as well as suggest that these organisms may be causative in asthma development. The existing data are not conclusive, but are suggestive enough to drive studies evaluating them as a possible mechanism in asthma pathogenesis. An animal model evaluating M. pneumoniae and C. pneumoniae would be ideal, but at present no model exists in which chronic infection with these organisms results in bronchial hyperresponsiveness. There is active work in this area, however. Alternative investigations include continued evaluation of these organisms by several modalities, including culture, serology, and PCR, along with evaluation of the host response. Many questions remain, but the ground is fertile for continued investigation.


Metformin exposure, maternal PCOS status and fetal venous liver circulation: A randomized, placebo-controlled study.
Metformin is prescribed to women with polycystic ovary syndrome (PCOS) to prevent pregnancy complications. Children exposed to metformin vs. placebo in utero, have increased head circumference at birth and are more overweight and obese at 8 years of age. Also, maternal PCOS-status seems to alter the long-term cardio-metabolic health of offspring. We hypothesized that the long-term effects of metformin-exposure and/or maternal PCOS may be mediated by circulatory adaptations during fetal life. This is a sub-study of a larger double-blinded, placebo-controlled trial, where women with PCOS were randomized to metformin (2g/day) or placebo in pregnancy, a total of 487 women. A sub-group of participants (N = 58) took part in this sub-study and had an extended ultrasound examination at gestational week 32, including blood flow velocity and diameter measurements of the umbilical vein (UV), the ductus venosus (DV) and the portal vein (PV). Blood flow volume was calculated and adjusted for estimated fetal weight (EFW) (normalized flow). Metformin exposed fetuses were compared to placebo exposed fetuses. Fetuses of mothers with PCOS (metformin [n = 30] and placebo [n = 28]) were compared to a low-risk reference population (N = 160) by z-score statistics. There was no difference in fetal liver flow between metformin vs. placebo-exposed fetuses. Fetuses of mothers with PCOS had higher EFW (0.63 [95% CI 0.44-0.83] p<0.001), lower normalized UV, DV, PV, and lower total venous liver blood flows than the reference population. Metformin during pregnancy did not affect fetal liver blood-flow. In our population, maternal PCOS-status was associated with reduced total venous liver blood-flow, which may explain altered growth and metabolism later in life.


The Red Queen theory of recombination hotspots.
Recombination hotspots are small chromosomal regions, where meiotic crossover events happen with high frequency. Recombination is initiated by a double-strand break (DSB) that requires the intervention of the molecular repair mechanism. The DSB repair mechanism may result in the exchange of homologous chromosomes (crossover) and the conversion of the allelic sequence that breaks into the one that does not break (biased gene conversion). Biased gene conversion results in a transmission advantage for the allele that does not break, thus preventing recombination and rendering recombination hotspots transient. How is it possible that recombination hotspots persist over evolutionary time (maintaining the average chromosomal crossover rate) when they are self-destructive? This fundamental question is known as the recombination hotspot paradox and has attracted much attention in recent years. Yet, that attention has not translated into a fully satisfactory answer. No existing model adequately explains all aspects of the recombination hotspot paradox. Here, we formulate an intragenomic conflict model resulting in Red Queen dynamics that fully accounts for all empirical observations regarding the molecular mechanisms of recombination hotspots, the nonrandom targeting of the recombination machinery to hotspots and the evolutionary dynamics of hotspot turnover.


Clopidogrel Therapy Discontinuation Following Drug Eluting Stent Implantation in Real World Practice in Israel.
Incidence and predictors of clopidogrel discontinuation after drug eluting stent (DES) implantation, in real world practice, are poorly known. Prospective study included all patients who underwent implantation of at least one DES between February 2006 and January 2007. Predictors of clopidogrel discontinuation were assessed by a multivariable analysis. In 269 patients, mean period for clopidogrel therapy was 13.2 ± 7.2 month. Twenty eight patients (10.4%) discontinued clolopidogrel prematurely (< 3 months). Early clopidogrel discontinuation was a predictor of late stent thrombosis (P = 0.005) and urgent target vessel revascularization (P = 0.05). There was a trend for higher cardiac mortality among that group (P = 0.07). By 12 months, 173 patients (64.3%) have discontinued clopidogrel therapy. The most frequent circumstance to stop clopidogrel before 12 months was recommendation of family physician. Patients that were followed by cardiologist were more encouraged to longer clopidogrel therapy. In multivariable analysis being non Jew (OR 19.2, 95% CI 2.4 to 142, P = 0.005), not followed by cardiologist (OR 4.7, 95% CI 1 to 23.1, P = 0.05) and lack of information regarding the importance of clopidogrel maintenance at discharge from hospital (OR 10.8, 95% CI 2.7 to 42.9, P = 0.001) were independent predictors of early clopidogrel discontinuation. Clopidogrel discontinuation, in real world practice is not unusual and related to poor outcome. Education for general physicians, clear instructions about the importance of antiplatelet maintenance at discharge and follow up by an expert cardiologist are opportunities to improve adherence do antiplatelet therapy following DES implantation.


Genetically detoxified mutants of heat-labile toxin from Escherichia coli are able to act as oral adjuvants.
Detoxified mutants of the Escherichia coli heat-labile toxin (LT) act as mucosal adjuvants to intranasally presented coadministered antigens. Here, we compare the adjuvant activity of a panel of detoxified derivatives of LT, using both intranasal (i.n.) and oral (p.o.) routes of administration. The mutants used as adjuvants varied in sensitivity to proteases and toxicity. With keyhole limpet hemocyanin (KLH) as the bystander antigen, the immune responses to i. n. immunizations were consistently higher than the equivalent p.o. -delivered proteins. LT-G192, a mutant which demonstrates a 10-fold reduction in toxicity in vitro, demonstrated wild-type adjuvant activity both i.n. and p.o., inducing similar titers of KLH specific antibody in the sera and immunoglobulin A in local mucosal secretions as wild-type LT. In line with previous data, the nontoxic holotoxoid LT-K63 induced intermediate immune responses in both the serum and mucosal secretions which were lower than those achieved with wild-type LT but at least 10-fold higher than those measured when the antigen was administered with LT-B. Although significant levels of local and systemic anti-KLH antibodies were induced following p.o. immunization with LT-K63, cellular proliferative responses to KLH was poor or undetectable. In contrast, LT and LT-G192 induced significant T-cell responses to KLH following p.o. immunization. These proliferating cells secreted both gamma interferon and interleukin-5, suggesting that the type of immune response induced following p.o. coimmunization with LT and purified protein is a mixed Th1/Th2 response.


Graphite pneumoconiosis. A review of etiologic and epidemiologic aspects.
Six hundred and five cases of graphite pneumoconiosis have been reported in the literature. In 39 cases the diagnosis was based on or supported by autopsy or lung biopsy results. Only 14 of the 39 cases were presented with relatively complete documentation as to details about dust exposure. Only one of these completely documented cases suggested that nearly pure graphite may cause graphite pneumoconiosis. Eleven experimental studies on animals dealt with the effect of graphite dust on various tissues, and nine with the effect on lung tissue. The three studies on the effect of pure or nearly pure graphite gave no unanimous conclusion. Five out of six studies dealing with graphite containing silica demonstrated fibrogenic effect on the lungs. Most of the 18 epidemiologic studies of workers exposed to graphite dust are invalid because they do not comply with ordinary methodologic requirements. The present state of knowledge does not exclude the possibility that analytically pure graphite may cause pneumoconiosis although the majority of the evidence indicates that pneumoconiosis is a mixed-dust type of lung reaction.


The results of nerve grafting in the wrist and hand.
To evaluate the usefulness of nerve grafting we studied 38 patients having 11 median, 7 ulnar, and 33 digital nerve grafts. Group funicular (interfascicular) grafting using magnification was performed in all patients. We followed 12 patients with 8 median and 5 ulnar nerve grafts for at least one year and 18 patients with 27 digital nerve grafts for at least six months. Medical Research Council criteria were used for evaluation of nerve function. Results in our patients and in previously reported patients having nerve grafting or repair were compared. Sensory function following ulnar nerve grafting was significantly better than that following nerve repair. Sensory function following median and digital nerve grafting was as good as that following nerve repair. Motor function following ulnar nerve grafting was as good as that following nerve repair. Previously reported patients having median nerve repairs or grafts had significantly better motor function than our patients.


Differential effect of prostaglandin synthetase inhibitor pretreatment on pentylenetetrazol-induced seizures in rat.
Release of prostaglandins (PGs) from brain tissue increases during experimentally-induced and spontaneous seizures. However, whether PGs or other arachidonic acid metabolites have a role in induction of seizures is still unclear. The effectiveness of pretreatment with PG synthetase inhibitors on pentylenetetrazol (PTZ)-induced models of epilepsy was investigated in free-moving rats with chronically-implanted supracortical electrodes. The effects on the electrocortical and motor manifestations of both a subconvulsive (30 mg.kg-1) and a convulsive (60 mg.kg-1) dose of PTZ were examined. Mefenamic acid (15 or 50 mg.kg-1), meclofenamic acid (15 or 50 mg.kg-1), ibuprofen (30 or 90 mg.kg-1) and paracetamol (300 or 450 mg.kg-1) delayed the onset of PTZ-induced convulsions. But mefenamic acid and meclofenamic acid also potentiated the excitatory effects of both subconvulsive and convulsive doses of PTZ. Indomethacin (3, 10 or 30 mg.kg-1) had no significant effect. The results suggest that the differential effects were produced by actions not related to the cyclo-oxygenase inhibition although the more consistent decrease of convulsive behavior may have resulted from inhibition of PG synthesis.


RFamide-related peptide-3(RFRP-3) receptor gene is expressed in mouse ovarian granulosa cells: potential role of RFRP-3 in steroidogenesis and apoptosis.
RFRP-3 is a functional ortholog of avian GnIH and regulates reproductive activities in the gonads of animals. However, the role of RFRP-3 in the function of ovarian granulosa cells in mice remains unclear. First, we detected the expression of the RFRP-3 receptor (GPR147) in the ovarian granulosa cells of mice. Second, the effect of RFRP-3 treatment on estradiol and progesterone secretions from granulosa cells was tested by ELISA. Meanwhile, the expression of genes and proteins regulating steroid hormone synthesis was respectively examined by qPCR and western blot. Furthermore, the effect of RFRP-3 treatment on the apoptosis of granulosa cells was analyzed. The results revealed that the GPR147 protein (a RFRP-3 receptor) was expressed in the ovarian granulosa cells of mice. Low and medium doses RFRP-3 treatment significantly reduced progesterone secretion in the granulosa cells (P < 0.05), while RFRP-3 suppressed p450scc, 3β-HSD, StAR, and FSHR expression in a non-dose-dependent manner. Moreover, RFRP-3 treatment might induce the apoptosis of granulosa cells. Additionally, low doses RFRP-3 significantly reduced p-ERK1/2 protein expression (P < 0.05) in the ovarian granulosa cells. We here, for the first time, confirmed that GPR147 was expressed in the ovarian granulosa cells of mice. Our findings suggested that and RFRP-3 regulates the granulosa cell function through the ERK signaling pathway, which will lay the foundation for uncovering molecular mechanisms by which RFRP-3 regulates follicle development in future.


Stress Introduction Rate Alters the Benefit of AcrAB-TolC Efflux Pumps.
Stress tolerance studies are typically conducted in an all-or-none fashion. However, in realistic settings-such as in clinical or metabolic engineering applications-cells may encounter stresses at different rates. Therefore, how cells tolerate stress may depend on its rate of appearance. To address this, we studied how the rate of stress introduction affects bacterial stress tolerance by focusing on a key stress response mechanism. Efflux pumps, such as AcrAB-TolC of Escherichia coli, are membrane transporters well known for the ability to export a wide variety of substrates, including antibiotics, signaling molecules, and biofuels. Although efflux pumps improve stress tolerance, pump overexpression can result in a substantial fitness cost to the cells. We hypothesized that the ideal pump expression level would involve a rate-dependent trade-off between the benefit of pumps and the cost of their expression. To test this, we evaluated the benefit of the AcrAB-TolC pump under different rates of stress introduction, including a step, a fast ramp, and a gradual ramp. Using two chemically diverse stresses, the antibiotic chloramphenicol and the jet biofuel precursor pinene, we assessed the benefit provided by the pumps. A mathematical model describing these effects predicted the benefit as a function of the rate of stress introduction. Our findings demonstrate that as the rate of introduction is lowered, stress response mechanisms provide a disproportionate benefit to pump-containing strains, allowing cells to survive beyond the original inhibitory concentrations.IMPORTANCE Efflux pumps are ubiquitous in nature and provide stress tolerance in the cells of species ranging from bacteria to mammals. Understanding how pumps provide tolerance has far-reaching implications for diverse fields, from medicine to biotechnology. Here, we investigated how the rate of stressor appearance impacts tolerance. We focused on two distinct substrates of AcrAB-TolC efflux pumps, the antibiotic chloramphenicol and the biofuel precursor pinene. Interestingly, tolerance is highly dependent on the rate of stress introduction. Therefore, it is important to consider not only the total quantity of a stressor but also the rate at which it is applied. The implications of this work are significant because environments are rarely static; antibiotic concentrations change during dosing, and metabolic engineering processes change with time.


The diversity and ecological roles of Penicillium in intertidal zones.
Members of the genus Penicillium are commonly isolated from various terrestrial and marine environments, and play an important ecological role as a decomposer. To gain insight into the ecological role of Penicillium in intertidal zones, we investigated the Penicillium diversity and community structure using a culture-dependent technique and a culture independent metagenomic approach using ITS (ITS-NGS) and partial β-tubulin (BenA-NGS) as targets. The obtained isolates were tested for halotolerance, enzyme activity, and polycyclic aromatic hydrocarbons (PAHs) degradation. A total of 96 Penicillium species were identified from the investigated intertidal zones. Although the BenA-NGS method was efficient for detecting Penicillium, some species were only detected using conventional isolation and/or the ITS-NGS method. The Penicillium community displayed a significant degree of variation relative to season (summer and winter) and seaside (western and southern coast). Many Penicillium species isolated in this study exhibited cellulase and protease activity, and/or degradation of PAHs. These findings support the important role of Penicillium in the intertidal zone for nutrient recycling and pollutant degradation.


Antifibrinolytics for heavy menstrual bleeding.
Heavy menstrual bleeding (HMB) is an important cause of ill health in women. Medical therapy, with the avoidance of possibly unnecessary surgery, is an attractive treatment option. A wide variety of medications are available to reduce heavy menstrual bleeding but there is considerable variation in practice and uncertainty about the most appropriate therapy. Plasminogen activators are a group of enzymes that cause fibrinolysis (the dissolution of clots). An increase in the levels of plasminogen activators has been found in the endometrium of women with heavy menstrual bleeding compared to those with normal menstrual loss. Plasminogen activator inhibitors (antifibrinolytic agents) have therefore been promoted as a treatment for heavy menstrual bleeding. There has been a reluctance to prescribe tranexamic acid due to possible side effects of the drugs such as an increased risk of thrombogenic disease (deep venous thrombosis). Long term studies in Sweden, however, have shown that the rate of incidence of thrombosis in women treated with tranexamic acid is comparable with the spontaneous frequency of thrombosis in women. To determine the effectiveness of antifibrinolytics in achieving a reduction in heavy menstrual bleeding. All studies which might describe randomised controlled trials of antifibrinolytic therapy for the treatment of heavy menstrual bleeding were obtained by electronic searches of MEDLINE 1966-1997, EMBASE 1980-1997 and the Cochrane Library. Companies producing antifibrinolytics and experts within the field were contacted for reference lists and information on unpublished trials. Randomised controlled trials in women of reproductive age treated with antifibrinolytic agents versus placebo, no treatment or any other medical (non-surgical) therapy for regular heavy menstrual bleeding within either the primary, family planning or specialist clinic settings. Women with post menopausal bleeding, intermenstrual bleeding, iatrogenic or pathological causes of heavy menstrual bleeding were excluded. Fifteen eligible trials were assessed by three reviewers and eight of these did not meet with the inclusion criteria. Of the seven remaining trials, four of these could be included within the meta-analysis. The remaining three trials had a crossover design and despite contacting the authors and appropriate companies, we were unable to extract the results in a format suitable to include these within the meta-analysis. However the results are included within the text of the review for discussion. Antifibrinolytic therapy compared to placebo showed a significant reduction in mean blood loss (WMD -94.0 [-151.4, -36.5]) and significant change in mean reduction of blood loss (WMD -110.2 [-146.5, -73.8]). This objective improvement was not mirrored by a patient perceived improvement in monthly menstrual blood loss (RR 2.5 [0.9, 7.3]) in the one study which recorded this outcome (~~ Edlund 1995~~). Antifibrinolytic agents were compared to only three other medical (non-surgical) therapies: mefenamic acid, norethisterone administered in the luteal phase and ethamsylate. In all instances, there was a significant reduction in mean blood loss (WMD -73.0 [-123.4, -22.6], WMD -111.0 [-178.5, -43.5] and (WMD -100 [-143.9, -56.1] respectively) and a strong, although non-significant trend in favour of tranexamic acid in the participants' perception of an improvement in menstrual blood loss. There were no significant differences in the frequency of reported side effects with tranexamic acid when compared to oral luteal phase progestogens (RR 0.4 [0.1, 1.2]) or withdrawal from treatment because of adverse events when compared with NSAIDs and ethamsylate when these treatments were used for heavy menstrual bleeding. Change in the quality of life measures, flooding and leakage and sex life, were significantly improved in the tranexamic acid group when compared to the oral progestagen group. These findings are based in most cases on only one trial. Antifibrinolytic therapy causes a greater reduction in objective measurements of heavy menstrual bleeding when compared to placebo or other medical therapies (NSAIDS, oral luteal phase progestagens and ethamsylate). This treatment is not associated with an increase in side effects compared to placebo, NSAIDS, oral luteal phase progestagens or ethamsylate. Flooding and leakage and sex life is significantly improved after tranexamic acid therapy when compared with oral luteal progestogens but no other measures of quality of life were assessed. No study has used resource cost as an outcome. There are no data available within randomised controlled trials which record the frequency of thromboembolic events.


Using genetic markers to directly estimate male selection gradients.
We present an analysis of Raphanus raphanistrum and simulations illustrating the utility of directly estimating male phenotypic selection gradients using genetic markers. The method offers a much more refined characterization of selection than attempting to assign paternity to individual progeny. Our analysis of R. raphanistrum reveals selection on remarkably fine features of floral morphology, including anther exsertion, that were opaque to previous approaches. The new results also undermine a previous conclusion that selection on wild radish floral morphology acts primarily through female fitness. Simulation results show that selection gradients on the order of beta = 0.1-0.2 can be readily detected with allozyme markers in moderate-sized (< 200 paternal individuals) populations. Highly polymorphic (e.g., microsatellite) markers will likely detect fine scale selection (beta < 0.1) in larger populations (> or = 400 individuals). Increased progeny sample size, by sampling either additional maternal families or more progeny per maternal parent, partly compensates for low exclusion probability. Increasing the number of possible fathers without changing progeny sample size decreases the ability to detect selection, especially at lower exclusion probabilities. Sampling only some male genotypes reduces the power to detect selection and biases (underestimates) the magnitude of the selection gradient estimate.


An Emerging Role for Sigma-1 Receptors in the Treatment of Developmental and Epileptic Encephalopathies.
Developmental and epileptic encephalopathies (DEEs) are complex conditions characterized primarily by seizures associated with neurodevelopmental and motor deficits. Recent evidence supports sigma-1 receptor modulation in both neuroprotection and antiseizure activity, suggesting that sigma-1 receptors may play a role in the pathogenesis of DEEs, and that targeting this receptor has the potential to positively impact both seizures and non-seizure outcomes in these disorders. Recent studies have demonstrated that the antiseizure medication fenfluramine, a serotonin-releasing drug that also acts as a positive modulator of sigma-1 receptors, reduces seizures and improves everyday executive functions (behavior, emotions, cognition) in patients with Dravet syndrome and Lennox-Gastaut syndrome. Here, we review the evidence for sigma-1 activity in reducing seizure frequency and promoting neuroprotection in the context of DEE pathophysiology and clinical presentation, using fenfluramine as a case example. Challenges and opportunities for future research include developing appropriate models for evaluating sigma-1 receptors in these syndromic epileptic conditions with multisystem involvement and complex clinical presentation.


Receptor-mediated tobacco toxicity: acceleration of sequential expression of alpha5 and alpha7 nicotinic receptor subunits in oral keratinocytes exposed to cigarette smoke.
Tobacco products and nicotine alter the cell cycle and lead to squamatization of oral keratinocytes (KCs) and squamous cell carcinoma. Activation of nicotinic acetylcholine receptors (nAChRs) elicits Ca(2+) influx that varies in magnitude between different nAChR subtypes. Normal differentiation of KCs is associated with sequential expression of the nAChR subtypes with increasing Ca(2+) permeability, such as alpha5-containing alpha3 nAChR and alpha7 nAChR. Exposure to environmental tobacco smoke (ETS) or an equivalent concentration of nicotine accelerated by severalfold the alpha5 and alpha7 expression in KCs, which could be abolished by mecamylamine and alpha-bungarotoxin with different efficacies, suggesting the following sequence of autoregulation of the expression of nAChR subtypes: alpha3(beta2/beta4) > alpha3(beta2/beta4)alpha5 > alpha7 > alpha7. This conjecture was corroborated by results of quantitative assays of subunit mRNA and protein levels, using nAChR-specific pharmacologic antagonists and small interfering RNAs. The genomic effects of ETS and nicotine involved the transcription factor GATA-2 that showed a multifold increase in quantity and activity in exposed KCs. Using protein kinase inhibitors and dominant negative and constitutively active constructs, we characterized the principal signaling cascades mediating a switch in the nAChR subtype. Cumulative results indicated that the alpha3(beta2/beta4) to alpha3(beta2/beta4)alpha5 nAChR transition predominantly involved protein kinase C, alpha3(beta2/beta4)alpha5 to alpha7 nAChR transition-Ca(2+)/calmodulin-dependent protein kinase II and p38 MAPK, and alpha7 self-up-regulation-the p38 MAPK/Akt pathway, and JAK-2. These results provide a mechanistic insight into the genomic effects of ETS and nicotine on KCs and characterize signaling pathways mediating autoregulation of stepwise overexpression of nAChR subtypes with increasing Ca(2+) permeability in exposed cells. These observations have salient clinical implications, because a switch in the nAChR subunit composition can bring about a corresponding switch in receptor function, leading to profound pathobiologic effects observed in KCs exposed to tobacco products.


Influence of an educational seminar on use of disease activity measurements by rheumatologists in treatment of rheumatoid arthritis.
To determine the variables underlying clinical decisions made by rheumatologists when treating patients with rheumatoid arthritis (RA), and to determine the effect of an educational seminar on the use of quantitative disease activity measurements in clinical practice in this population of physicians. Practicing rheumatologists were surveyed on the variables affecting their clinical management of patients with RA by questionnaire. Physicians were divided into 2 groups: the first comprised attenders (Group A) to an educational seminar in the use of the quantitative disease activity measurements in patient management, while the second group comprised nonattenders (Group NA). Both groups were surveyed on their practice behavior before (Survey 1) and 2 to 3 months after (Survey 2) the seminar. Fifty-two rheumatologists in clinical practice from across the US completed and returned 364 surveys. A significantly greater number of rheumatologists in Group A reported use of disease activity measures following the training seminar (Survey 2), compared to their use pre-meeting and compared to Group NA (p < 0.0001). Our results support employment of an educational seminar on the use of disease activity measurements to increase the use of these quantitative measures in rheumatologic practice.


Hepatic stellate cells: a target for the treatment of liver fibrosis.
Hepatic fibrosis is a wound-healing process that occurs when the liver is injured chronically. Hepatic stellate cells (HSC) are responsible for the excess production of extracellular matrix (ECM) components. The activation of HSC, a key issue in the pathogenesis of hepatic fibrosis, is mediated by various cytokines and reactive oxygen species released from the damaged hepatocytes and activated Kupffer cells. Therefore, inhibition of HSC activation and its related subsequent events, such as increased production of ECM components and enhanced proliferation, are crucial goals for intervention in the hepatic fibrogenesis cascade. This is especially true when the etiology is unknown or there is no established therapy for the cause of the chronic injury. This review explores the rationale for choosing HSC as a target for the pharmacological, molecular, and other novel therapeutics for hepatic fibrosis. One focus of this review is the inhibition of two cytokines, transforming growth factor-beta and platelet-derived growth factor, which are important in hepatic fibrogenesis. A number of new agents, such as Chinese herbal recipes and herbal extracts, silymarin, S-adenosyl-L-methionine, polyenylphosphatidylcholine, and pentoxifylline are also discussed.


[Gabapentin (Neurontin) and cancer pain: a pilot study].
Gabapentin (GBP) is a new antiepileptic agent with an original spectrum of activity. Its mechanism of action has not yet been fully elucidated but appears not to involve binding to GABA receptors despite being a structural analogue of GABA and is distinct from tricyclic antidepressants (TCAs). It has been shown to modulate high threshold calcium currents in brain neurons. As some other anticonvulsants, GBP has been recently proposed for the treatment of noncancer neuropathic pain like diabetic neuropathy and post herpetic neuralgia (double blind studies with placebo). We prospectively followed 20 cancer patients with advanced disease suffering from neuropathic pain. All were already treated for their pain syndrome. We started with 300 mg of GBP given orally in order to reach a dose of 900 mg on D3. All coanalgesics were stopped before entering the study. The only relevant side effect due to GBP was somnolence, otherwise time limited. GBP treatment was associated with a decrease of opioids doses in 9 patients and a decrease of VAS for pain intensity in all cases. Furthermore, the need of rescue doses decreased in all cases but 2. GBP appears to be one of the most effective drugs for the treatment of neuropathic pain. It is well tolerated and its effectiveness appears shortly after its administration. A synergistic action with opioids is suggested. Despite the small number of patients, our study suggests that GBP could be a treatment of neuropathic pain in cancer. Comparative trials should be performed with other neuropathic pain drugs including TCAs and antiepileptic drugs, especially carbamazepine.


The Thermoelectric Properties of n-Type Bismuth Telluride: Bismuth Selenide Alloys Bi2Te3-x Se x.
Alloying bismuth telluride with antimony telluride and bismuth selenide for p- and n-type materials, respectively, improves the thermoelectric quality factor for use in room temperature modules. As the electronic and thermal transports can vary substantially, the alloy composition is a key engineering parameter. The n-type Bi2Te3- x Se x alloy lags its p-type counterpart in thermoelectric performance and does not lend itself as readily to simple transport modeling which complicates engineering. Combining literature data with recent results across the entire alloy composition range, the complex electronic structure dynamics and trends in lattice thermal conductivity are explored. Spin-orbit interaction plays a critical role in determining the position and degeneracy of the various conduction band minima. This behavior is incorporated into a two-band effective mass model to estimate the transport parameters in each band. An alloy scattering model is utilized to demonstrate how phonon scattering behaves differently on either side of the intermediate ordered compound Bi2Te2Se due to chalcogen site occupancy preference. The parametrization of the electronic and thermal transports presented can be used in future optimization efforts.


Netting neutrophils are major inducers of type I IFN production in pediatric systemic lupus erythematosus.
Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by a breakdown of tolerance to nuclear antigens and the development of immune complexes. Genomic approaches have shown that human SLE leukocytes homogeneously express type I interferon (IFN)-induced and neutrophil-related transcripts. Increased production and/or bioavailability of IFN-α and associated alterations in dendritic cell (DC) homeostasis have been linked to lupus pathogenesis. Although neutrophils have long been shown to be associated with lupus, their potential role in disease pathogenesis remains elusive. Here, we show that mature SLE neutrophils are primed in vivo by type I IFN and die upon exposure to SLE-derived anti-ribonucleoprotein antibodies, releasing neutrophil extracellular traps (NETs). SLE NETs contain DNA as well as large amounts of LL37 and HMGB1, neutrophil proteins that facilitate the uptake and recognition of mammalian DNA by plasmacytoid DCs (pDCs). Indeed, SLE NETs activate pDCs to produce high levels of IFN-α in a DNA- and TLR9 (Toll-like receptor 9)-dependent manner. Our results reveal an unsuspected role for neutrophils in SLE pathogenesis and identify a novel link between nucleic acid-recognizing antibodies and type I IFN production in this disease.


Motor neuron disease associated with carcinoma. A report of 2 cases.
Paraneoplastic complications are obscure and difficult to understand. The association of motor neuron disease and carcinoma may sometimes be more than coincidental, and 2 cases are described. One patient had motor neuron disease, limbic encephalitis (a recognized paraneoplastic disorder) and carcinoma of the oesophagus; the other had motor neuron disease and adenocarcinoma of the rectum. In the elderly male with motor neuron disease simple screening tests to exclude lymphoma and carcinoma of the lung, bowel and genito-urinary tract are advocated.


Improving validity of informed consent for biomedical research in Zambia using a laboratory exposure intervention.
Complex biomedical research can lead to disquiet in communities with limited exposure to scientific discussions, leading to rumours or to high drop-out rates. We set out to test an intervention designed to address apprehensions commonly encountered in a community where literacy is uncommon, and where complex biomedical research has been conducted for over a decade. We aimed to determine if it could improve the validity of consent. Data were collected using focus group discussions, key informant interviews and observations. We designed an intervention that exposed participants to a detailed demonstration of laboratory processes. Each group was interviewed twice in a day, before and after exposure to the intervention in order to assess changes in their views. Factors that motivated people to participate in invasive biomedical research included a desire to stay healthy because of the screening during the recruitment process, regular advice from doctors, free medical services, and trust in the researchers. Inhibiting factors were limited knowledge about samples taken from their bodies during endoscopic procedures, the impact of endoscopy on the function of internal organs, and concerns about the use of biomedical samples. The belief that blood can be used for Satanic practices also created insecurities about drawing of blood samples. Further inhibiting factors included a fear of being labelled as HIV positive if known to consult heath workers repeatedly, and gender inequality. Concerns about the use and storage of blood and tissue samples were overcome by a laboratory exposure intervention. Selecting a group of members from target community and engaging them in a laboratory exposure intervention could be a useful tool for enhancing specific aspects of consent for biomedical research. Further work is needed to determine the extent to which improved understanding permeates beyond the immediate group participating in the intervention.


Testing for antireticulin antibodies in patients with celiac disease is obsolete: a review of recommendations for serologic screening and the literature.
Celiac disease (CD) is an autoimmune disorder that occurs in genetically susceptible individuals of all ages and is triggered by immune response to gluten and related proteins. The disease is characterized by the presence of HLA-DQ2 and/or -DQ8 haplotypes, diverse clinical manifestations, gluten-sensitive enteropathy, and production of several autoantibodies of which endomysial, tissue transglutaminase, and deamidated gliadin peptide antibodies are considered specific. Although antireticulin antibodies (ARA) have historically been used in the evaluation of CD, these assays lack optimal sensitivities and specificities for routine diagnostic use. This minireview highlights the advances in CD-specific serologic testing and the rationale for eliminating ARA from CD evaluation consistent with recommendations for diagnosis.


Evaluation of the Effectiveness of Psychoeducation Given to Children with Parental Psychiatric Disorders.
This study was aimed to determine the effectiveness of psychoeducation program which was developed to improve the coping skills and to increase the psychological resistance of 12-18 years children/adolescents whose parents' have psychiatric disorders. The study was carried out between May 2018 - November 2019 with children of patients who applied to the outpatient clinic and inpatient treatment at Istanbul University - Cerrahpasa, Cerrahpasa Faculty of Medicine Department of Psychiatry. Total of 40 children (experiment = 20 and control = 20), participated in the study. Data were collected using Personal Information Form, Adolescent Psychological Resilience Scale and Kidcope. The mean age of the children/adolescents in the experimental group was 14.05 ± 1.90, 50% was male and the mean age of the control group was 15.35 ± 2.08, and 55% were male. It was determined that the mean duration of psychiatric disorders were 11.50 ± 6.39 years and 40% of the psychiatric diagnosis was schizophrenia in the experimental group, and the mean duration psychiatric disorders were 6.10 ± 5.16 years and 30% of the psychiatric diagnosis was a depression in the control group. After psychoeducation, the total scores of scales in the experimental group showed an increase compared to the control group and a significant difference in a statistical context. It was found that children/adolescents participating in the psychoeducation program had increased psychological resilience levels and improved coping skills. This result shows that the psychoeducation program for children/adolescents with psychiatric disorders in their parents is effective.


Junctional cleft [Ca²⁺]i measurements using novel cleft-targeted Ca²⁺ sensors.
Intracellular Ca(2+) concentration ([Ca(2+)]i) is regulated and signals differently in various subcellular microdomains, which greatly enhances its second messenger versatility. In the heart, sarcoplasmic reticulum Ca(2+) release and signaling are controlled by local [Ca(2+)]i in the junctional cleft ([Ca(2+)]Cleft), the small space between sarcolemma and junctional sarcoplasmic reticulum. However, methods to measure [Ca(2+)]Cleft directly are needed. To construct novel sensors that allow direct measurement of [Ca(2+)]Cleft. We constructed cleft-targeted [Ca(2+)] sensors by fusing Ca(2+)-sensor GCaMP2.2 and a new lower Ca(2+)-affinity variant GCaMP2.2Low to FKBP12.6, which binds with high affinity and selectivity to ryanodine receptors. The fluorescence pattern, affinity for ryanodine receptors, and competition by untagged FKBP12.6 demonstrated that FKBP12.6-tagged sensors are positioned to measure local [Ca(2+)]Cleft in adult rat myocytes. Using GCaMP2.2Low-FKBP12.6, we showed that [Ca(2+)]Cleft reaches higher levels with faster kinetics than global [Ca(2+)]i during excitation-contraction coupling. Diastolic sarcoplasmic reticulum Ca(2+) leak or sarcolemmal Ca(2+) entry may raise local [Ca(2+)]Cleft above bulk cytosolic [Ca(2+)]i ([Ca(2+)]Bulk), an effect that may contribute to triggered arrhythmias and even transcriptional regulation. We measured this diastolic standing [Ca(2+)]Cleft-[Ca(2+)]Bulk gradient with GCaMP2.2-FKBP12.6 versus GCaMP2.2, using [Ca(2+)] measured without gradients as a reference point. This diastolic difference ([Ca(2+)]Cleft=194 nmol/L versus [Ca(2+)]Bulk=100 nmol/L) is dictated mainly by the sarcoplasmic reticulum Ca(2+) leak rather than sarcolemmal Ca(2+) flux. We have developed junctional cleft-targeted sensors to measure [Ca(2+)]Cleft versus [Ca(2+)]Bulk and demonstrated dynamic differences during electric excitation and a standing diastolic [Ca(2+)]i gradient, which could influence local Ca(2+)-dependent signaling within the junctional cleft.


Impact of angiotensin-converting enzyme gene polymorphism on proteinuria and arterial hypertension.
Proteinuria is the hallmark of renal disease. In essential hypertension the onset of de novo proteinuria is associated with faster rate of progression of disease. Some authors have suggested that the DD genotype of the angiotensin-converting enzyme (ACE) gene would be an adverse renal prognosis factor. It may also have different effects on the reduction of proteinuria by ACE inhibitors in patients with proteinuria. Observations on the association between the ACE gene polymorphism and hypertension have been inconsistent, which might be due to ethnic and geographical variations. In this study was to investigated the relationship between ACE gene polymorphism and antiproteinuric effect of ACE inhibitors (ramipril) and to evaluate the possible association between I/D polymorphism and hypertension. We recruited 66 hypertensive patients (male 42, female 24) with overt proteinuria (urinary protein excretion over 500 mg/day). Patients were classified into three groups in accordance with ACE genotypes (17 DD; 35 ID; 14 II). They were treated with ramipril and prospectively followed up for one year. Various clinical parameters including age, body mass index (BMI), 24-h urine protein, creatinine, creatinine clearance (Ccr), systolic and diastolic blood pressure (SBP and DBP), mean arterial pressure (MAP) were measured in the pre- and post-treatment periods. The ACE gene insertion/deletion(I/D) polymorphisms in intron 16 were determined by PCR. Results showed that there were no significant differences in the clinical parameters such as age, gender, serum creatinine, Ccr, SBP, DBP, MAP, and daily urinary excretion of protein among three groups (P > 0.05). ID genotype patients were found to have lower BMI (p = 0.031). ACE inhibition significantly reduced proteinuria in all genotype groups (p < 0.05). The percentage reductions of 24-h urinary excretion of protein were significantly different between the genotype groups (p = 0.042) and for DD genotype were significantly greater than in ID (79.2 +/- 28.9% vs 49.2 +/- 64.8%, P = 0.015). The slope of SBP was the main factor related to the slope of the percentage reduction of proteinuria, however, a significant negative correlation coefficient between these parameters was found (rs = -0.382, p = 0.002). We failed to find significant difference in outcomes of treatments with ACE inhibitor between male and female according the I/D polymorphism of the ACE gene. D allele in the ACE genotype could be a useful genetic marker with important clinical, therapeutic and prognostic implications in recognizing patients with proteinuria that are at greater risk of renal damage.


Isolation and characterization of a human liver cDNA as a candidate gene for Wilson disease.
The putative copper and ATP-binding domains of the human Menkes disease gene were used as probes to screen a human liver cDNA library at reduced stringency. Sixty-five clones which remained positive after tertiary screening were subcloned and sequenced. One of these cDNA clones contains an open reading frame with 65% amino acid homology to the Menkes protein. Southern blot analysis localizes this cDNA to the region of the Wilson disease locus on chromosome 13. This cDNA detects a 7.5 kB transcript which is present in human liver and cell lines devoid of the Menkes transcript and which is absent in liver from a patient with Wilson disease. These data suggest that this cDNA is a candidate gene for Wilson disease and that the protein encoded at this locus is a member of the P-type ATPase family.


Hydrogen peroxide-mediated isoniazid activation catalyzed by Mycobacterium tuberculosis catalase-peroxidase (KatG) and its S315T mutant.
Inhibition of the enzyme Mycobacterium tuberculosis InhA (enoyl-acyl carrier protein reductase) due to formation of an isonicotinoyl-NAD adduct (IN-NAD) from isoniazid (INH) and nicotinamide adenine dinucleotide cofactor is considered central to the mode of action of INH, a first-line treatment for tuberculosis infection. INH action against mycobacteria requires catalase-peroxidase (KatG) function, and IN-NAD adduct formation is catalyzed in vitro by M. tuberculosis KatG under a variety of conditions, yet a physiologically relevant approach to the process has not emerged that allows scrutiny of the mechanism and the origins of INH resistance in the most prevalent drug-resistant strain bearing KatG[S315T]. In this report, we describe how hydrogen peroxide, delivered at very low concentrations to ferric KatG, leads to efficient inhibition of InhA due to formation of the IN-NAD adduct. The rate of adduct formation mediated by wild-type KatG was about 20-fold greater than by the isoniazid-resistant KatG[S315T] mutant under optimal conditions (H2O2 supplied along with NAD+ and INH). Slow adduct formation also occurs starting with NADH and INH, in the presence of KatG even in the absence of added peroxide, due to endogenous peroxide. The poor efficiency of the KatG[S315T] mutant can be enhanced merely by increasing the concentration of INH, consistent with this enzyme's reduced affinity for INH binding to the resting enzyme and the catalytically competent enzyme intermediate (Compound I). Origins of drug resistance in the KatG[S315T] mutant enzyme are analyzed at the structural level through examination of the three-dimensional X-ray crystal structure of the mutant enzyme.


Alcohol abstinence, non-hazardous use and hazardous use a decade after alcohol-related hospitalization: registry data linked to population-based representative postal surveys.
Although there is evident association between alcohol-related hospitalization and alcohol use, the relationship has not been well examined. This study analyzed the extent of alcohol abstinence, non-hazardous use and hazardous use among people who had experienced alcohol-related hospitalization during the preceding decade. Registry data concerning alcohol-related hospitalizations between 1996 and 2007 were linked to two representative surveys, in 2006 and 2007, of residents of Stockholm County. Relevant contrasts were modeled, using logistic regression, in the pooled sample (n = 54 955). Ages were 23-84 years at follow-up. Among persons previously hospitalized (n = 576), half reported non-hazardous use. Non-hazardous use was less prevalent than in the general population--and the extent of non-hazardous use did not change over time following hospitalization. There were no significant age differences, but non-hazardous use was less frequent among people with repeated episodes of care. One in six was abstinent. Abstinence was more common among the old, while hazardous use (exceeding 14 drinks per week for men, and 9 drinks per week for women) decreased with age. Abstinence also increased over time; among persons hospitalized ten years ago, the abstinence rate was twice that of the general population. Associations with hazardous use over time were less conclusive. Hazardous use among those previously hospitalized decreased over time in one sample but not in the other. After pooling the data, there were indications of a decrease over time following hospitalization, but more prevalent hazardous use than in the general population. Following alcohol-related hospitalization, abstinence increased, and there was no evidence of regression towards the mean, i.e., towards non-hazardous use. Abstinence was also more widespread among previously hospitalized persons of older ages. With advancing age, changing hazardous alcohol habits among previously hospitalized appears to yield a trend towards promotion of abstinence.


Synergie between molecular imprinted polymer based on solid-phase extraction and quartz crystal microbalance technique for 8-OHdG sensing.
Recently, the 8-hydroxy-2'-deoxyguanosine (8-OHdG) has been used as a marker to determine the oxidative stress. There is no any cheap and easy determination method based on chips and sensor systems for the determination of 8-OHdG. In this study, we have proposed imprinting methods for 8-OHdG recognition and determination using methacryloylamidohistidine-platinum(II) [MAH-Pt(II)] as a new metal-chelating monomer. The study includes the solid-phase extraction (SPE) of blood sample by a new 8-OHdG imprinted sorbent and the measurement of binding interaction of 8-OHdG imprinted quartz crystal microbalance (QCM) sensor via ligand interaction. 8-OHdG imprinted sorbent has prepared by bulk polymerization of MAH-Pt(II) and N-N'-methylenebisacrylamide. 8-OHdG imprinted sensor has prepared on a QCM chip coating the thiol pretreated Au electrode. At the end of these steps, a thin molecular imprinted polymer (MIP) film for the detection of 8-OHdG has developed and analytical performance of QCM sensor which has prepared using MIP was investigated. The affinity constant (K(a)) for 8-OHdG using MAH-Pt-based thin film has determined by using the Scatchard method. The average percentage recovery of 8-OHdG from plasma samples was found as 80% by using of 8-OHdG imprinted SPE material. At the last step, 8-OHdG level in several blood plasma has been determined by this improved QCM sensor. The obtained results confirmed that the 8-OHdG level in cancer patient's blood was significantly higher than in general subjects.


Interleukin (IL)-9/IL-9R axis drives γδ T cells activation in psoriatic arthritis patients.
Cytokines such as tumour necrosis factor (TNF)-α, interleukin (IL)-12, interferon (IFN)-γ, IL-23 and, more recently, IL-9, have been implicated in the initiation/maintenance of inflammation in psoriasis and psoriatic arthritis (PsA). In the present study we aimed to characterize the role of γδ T cells in peripheral blood and synovial fluid of PsA patients and to investigate their response to in-vitro stimulation with antigen or cytokines (IL-9 and IL-23). γδ T cells isolated from peripheral blood mononuclear cells and synovial fluid were analysed by flow cytometry to evaluate the phenotype and cytokine production. IL-23R and IL-9R gene expression were also evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Peripheral blood mononuclear cells (PBMC), sorted γδ T cells and γδ cell lines were also stimulated in vitro with isopentenyl pyrophosphate (IPP), recombinant IL-9 or recombinant IL-23. Our results show an expansion of γδ T cells with a predominant effector memory phenotype in peripheral blood and synovium of untreated PsA patients, which reverses significantly after treatment with anti-TNF-α or anti-IL-12/IL-23R monoclonal antibodies (mAbs). Moreover, in PsA patients γδ T cells activation is driven prevalently by IL-9/IL-9R interaction, and not only by IL-23/IL-23R. Together these findings indicate γδ T cells and IL-9 as new players in the pathogenesis of PsA.


The short history of gastroenterology.
In this paper research on the stomach and bowel physiology is presented in a historical perspective. The author tries to show how digestive processes were interpreted by the ancients and how they tried to adjust them to the dominating humoral theory of disease. It is pointed out that the breakthrough which created a new way of understanding of the function of the digestive system was made by Andreas Vesalius and his modern model of anatomy. The meaning of acceptance of chemical processes in digestion by iatrochemics representatives in XVII century is shown. Physiological research in XIX century, which decided about a rapid development of physiology, especially the physiology of the gastrointestinal tract, is discussed. Experiments were performed by all main representatives of this discipline: Claude Bernard, Jan Ewangelista Purkyne, Rudolph Heidenhain and especially Ivan Pavlov, who, thanks to the discoveries in the secretion physiology, explained basic functions of the central nervous system. The XX century was dominated by the research showing the important role of the endocrine system and biological agents in the regulation of secretion and motility of the digestive system. The following discoveries are discussed: Ernest Sterling (secretin), John Edkins (gastrin) and André Latarjet and Lester Dragstedt (acetylcholine). It is underlined that Polish scientists play an important role in the development of the gastroenterological science--among others, Walery Jaworski, who made a historical suggestion about the role of the spiral bacteria in etiopathogenesis of the peptic ulcer, Leon Popielski, who stated the stimulating influence of histamine on the stomach acid secretion, Julian Walawski, who discovered enterogastrons--hormones decreasing secretion. As a supplement, there is the list of achievements in the field of the physiology and pathology of the gastrointestinal tract awarded with Nobel Prize and the list of most important Polish papers in this field.


Recurrent carotid body tumors: embolization as a treatment option.
We report a clinical case where transcatheter embolization was selected as the primary treatment for a large recurrent carotid body tumor A 55 year-old female presented with a painful left cervical mass, with progressive growth, for the past 12 months. She complained of jaw stiffness, odynophagia and dysphonia. She had a former history of bilateral carotid body tumor resection. The patient underwent carotid ultrasound examination that showed a recurrent left carotid body tumor with 7x5 cm in dimension, and occlusion of the left internal carotid artery. Magnetic resonance imaging confirmed the presence of a Shamblin type III tumor. The patient underwent transcatheter embolization of the tumor with 300-500 µm and 500-700 µm Bead-Block®. At 1 year of follow-up, the patient was found asymptomatic. The carotid body is located at the bifurcation of the common carotid artery. With increased size, carotid body tumors can induce significant symptoms and are usually detected by clinical examination. Confirmation of diagnosis is usually given by vascular ultrasound. For highly symptomatic, recurrent and frequently unresectable tumors--in patients unfit for surgery--transcatheter embolization can also be used as an effective palliative treatment.


Branched microtubule nucleation and dynein transport organize RanGTP asters in Xenopus laevis egg extract.
Chromosome segregation relies on the correct assembly of a bipolar spindle. Spindle pole self-organization requires dynein-dependent microtubule (MT) transport along other MTs. However, during M-phase RanGTP triggers MT nucleation and branching generating polarized arrays with nonastral organization in which MT minus ends are linked to the sides of other MTs. This raises the question of how branched-MT nucleation and dynein-mediated transport cooperate to organize the spindle poles. Here, we used RanGTP-dependent MT aster formation in Xenopus laevis (X. laevis) egg extract to study the interplay between these two seemingly conflicting organizing principles. Using temporally controlled perturbations of MT nucleation and dynein activity, we found that branched MTs are not static but instead dynamically redistribute over time as poles self-organize. Our experimental data together with computer simulations suggest a model where dynein together with dynactin and NuMA directly pulls and move branched MT minus ends toward other MT minus ends.


Sustaining care for a parent with dementia: an indefinite and intertwined process.
This study aimed to understand how adult children sustain caring for persons with dementia (PwDs) within their family and formal care contexts in Canada. Half-day focus groups were conducted with adult daughters and adult sons in Toronto, Canada. Using constructivist grounded theory, we examined both substantive concepts and group dynamics. Sustaining care was interpreted as an indefinite process with three intertwined themes: reproducing care demands and dependency, enacting and affirming values, and "flying blind" in how and how long to sustain caring (i.e., responding to immediate needs with limited foresight). Family values and relationships, mistrust toward the institutional and home care systems, and obscured care foresight influenced care decisions and challenged participants in balancing their parents' needs with their own. Positive and negative aspects of care were found to influence one another. The implications of these findings for research and policy are discussed.


Effect of intracellular pH on spontaneous Ca2+ sparks in rat ventricular myocytes.
1. A fall of intracellular pH (pHi) typically depresses cardiac contractility. Among the many mechanisms underlying this depression, an inhibitory effect of acidosis upon the sarcoplasmic reticulum (SR) Ca2+ release channel has been predicted, but not so far demonstrated in the intact cardiac myocyte. In the present work, pHi was manipulated experimentally while confocal imaging was used to record spontaneous 'Ca2+ sparks' (local SR Ca2+ release events) in rat isolated myocytes loaded with the fluorescent Ca2+ indicator fluo-3. In other experiments, whole cell (global) pHi or [Ca2+]i was measured by microfluorimetry (using, respectively, intracellular carboxy SNARF-1 and indo-1). 2. Reducing pHi (i) increased whole cell intracellular [Ca2+] transients induced either electrically or by addition of caffeine, whereas (ii) it decreased spontaneous Ca2+ spark frequency. Conversely, raising pHi increased spontaneous Ca2+ spark frequency. 3. Blocking sarcolemmal Ca2+ influx with 10 mM Ni2+, or reducing external pH by 1.0 unit, had no effect on the pHi-dependent changes in spontaneous Ca2+ spark frequency. 4. Decreasing pHi over the range 7.78-7.20, decreased Ca2+ spark frequency exponentially as a function of pHi, with frequency declining by approximately 33 % for a 0.2 unit fall in pHi. In contrast, over the same pHi range, Ca2+ spark amplitude was unaffected. Intracellular acidosis produced a slight slowing of Ca2+ spark relaxation. 5. The results indicate that, in the intact myocyte, a reduced pHi decreases the probability of opening of the SR Ca2+ release channel. This phenomenon may contribute to the negative inotropic effects of acidosis.


Evaluation of dexmedetomidine-midazolam sedation in budgerigars (Melopsittacus undulatus).
To evaluate the sedative effects of IM administration of a high or low dose of dexmedetomidine in combination with midazolam in budgerigars (Melopsittacus undulatus). 20 healthy adult budgerigars. In a prospective, randomized, blinded study, birds were sedated with a high dose (HD; 0.04 mg/kg, IM; n = 10) or low dose (LD; 0.01 mg/kg, IM; 10) of dexmedetomidine in combination with midazolam (3 mg/kg, IM). Twenty minutes later, atipamezole (0.4 mg/kg [HD group] or 0.1 mg/kg [LD group], IM) and flumazenil (0.1 mg/kg, IM) were administered for reversal of sedation. Times to first effect and to sternal recumbency after administration of the sedatives and times to standing and eating after administration of the antagonists did not differ between groups. Most birds (9/10 in the HD group and 7/10 in the LD group) lost the righting response by 10 minutes after sedative administration, and the peak effect for radiographic positioning was by 15 minutes. Although it was not clinically relevant, most birds showed mild resedation by 60 minutes after administration of the reversal agents. There was no significant cardiorespiratory compromise detected with either protocol. Dexmedetomidine-midazolam can safely and effectively provide a dose-dependent level of sedation in healthy budgerigars. The HD protocol is recommended for radiographic positioning, as it allows for a more reliable, deeper plane of sedation.


Distance measurements via the morphogen gradient of Bicoid in Drosophila embryos.
Patterning along the anterior-posterior (A-P) axis in Drosophila embryos is instructed by the morphogen gradient of Bicoid (Bcd). Despite extensive studies of this morphogen, how embryo geometry may affect gradient formation and target responses has not been investigated experimentally. In this report, we systematically compare the Bcd gradient profiles and its target expression patterns on the dorsal and ventral sides of the embryo. Our results support a hypothesis that proper distance measurement and the encoded positional information of the Bcd gradient are along the perimeter of the embryo. Our results also reveal that the dorsal and ventral sides of the embryo have a fundamentally similar relationship between Bcd and its target Hunchback (Hb), suggesting that Hb expression properties on the two sides of the embryo can be directly traced to Bcd gradient properties. Our 3-D simulation studies show that a curvature difference between the two sides of an embryo is sufficient to generate Bcd gradient properties that are consistent with experimental observations. The findings described in this report provide a first quantitative, experimental evaluation of embryo geometry on Bcd gradient formation and target responses. They demonstrate that the physical features of an embryo, such as its shape, are integral to how pattern is formed.


Isolation and characterization of two group A rotaviruses with unusual genome profiles.
Analysis of the genomic dsRNA of rotaviruses isolated from calves with subclinical infections has revealed eight calves excreting group A viruses with unusual genome profiles. Two of these virus strains, C7-176 and C7-183, were grown and cloned by plaque purification in cell culture. Examination of their genome profiles after cloning showed that the unusual pattern had been retained. They were without RNA segment 11 but had an extra band (6a) migrating between segments 6 and 7. However, they contained the triplet of segments 7, 8 and 9, of similar size, which is characteristic of group A rotaviruses. The number and mol. wt. of the intracellular polypeptides induced by these viruses were similar to those of the bovine group A rotavirus UK strain. Analysis of the RNA transcripts produced by the transcription in vitro of purified C7-183 virus showed that segment 6a produced a large RNA transcript of corresponding size. After isolation, this transcript was translated in a rabbit reticulocyte lysate preparation and yielded a single polypeptide, vp11, equivalent to the product of segment 11 of rotaviruses with typical genome profiles.


Durability of immune response against omicron BA.2 and BA.4/5 and T cell responses after boosting with mRNA and adenoviral vector-based vaccines following heterologous CoronaVac/ChAdOx-1nCov-19 vaccination.
Heterologous vaccination with inactivated vaccine followed by adenoviral vector-based vaccine has shown superiority in enhancing immune response compared to homologous primary series. However, data comparing immunity decline after a third booster following heterologous CoronaVac/ChAdOx-1nCov-19 has been limited. Here, we assessed neutralizing activity against omicron variant and T cell response at 3 months monitoring in 96 individuals who received ChAdOx-1nCov-19, BNT162b2, or mRNA-1273 as a third dose following heterologous CoronaVac/ChAdOx-1nCov-19. Comparing the antibody levels at 3 and 1 month(s) after the third booster, the results showed a persistence of anti-RBD IgG in all vaccine regimens, with the IgG level waning slower in the ChAdOx-1nCov-19 boosted group (geometric mean ratio (GMR): 0.64 (95%CI: 0.59-0.70)) compared to the BNT162b2 (0.34 (95%CI:0.31-0.38)) and mRNA-1273 boosted groups (0.32 (95%CI: 0.29-0.36)). Neutralizing activity against omicron BA.2 and BA.4/5 dropped by 1.2 to 1.5-fold but remained detectable, with the highest level observed in the mRNA-1273 group, followed by BNT162b2 and ChAdOx-1nCov-19 groups, respectively. Furthermore, the number of individuals with T cell reactivity decreased in BNT162b2 and mRNA-1273 groups, while it increased in ChAdOx-1nCov-19 group at 3-month post-boost compared to 1 month. Data on the durability of immune response could help comprehensively optimize the booster vaccine strategy.


An electrophoretic DNA extraction device using a nanofilter for molecular diagnosis of pathogens.
Rapid and efficient nucleic acid (NA) extraction and concentration are required for point-of-care analysis in order to prevent an epidemic/pandemic disease outbreak. Typical silica-based NA extraction methods have limitations such as being time-consuming, requiring human intervention, and resulting in a low recovery yield. In this study, we have developed a pathogenic DNA extraction device based on electrokinetic separation incorporated with a silicon nitride (SiNx) nanofilter, which expedites the DNA extraction procedure with advantages of being convenient, efficient, and inexpensive. This DNA extraction device consists of a computer numerical control (CNC) milled-Teflon gadget with a cis-chamber as a cell lysate reservoir and a trans-chamber as a elution solution reservoir, with the SiNx nanofilter being inserted between the two chambers. The SiNx nanofilter was fabricated using a photolithographic method in conjunction with nanoimprinting. Approximately 7.2 million nanopores of 220 nm diameter were located at the center of the nanofilter. When a DC electric field is applied through the nanopores, DNA is transferred from the cis-chamber to the trans-chamber to isolate the DNA from the cell debris. To demonstrate the DNA extraction performance, we measured the absorbances at 260 and 280 nm and performed a real-time polymerase chain reaction (real-time PCR) using the recovered DNA to verify its feasibility for downstream genetic analysis. Moreover, the DNA extraction device was successfully operated using a 1.5 V alkaline battery, which verifies the portability of the device for point-of-care testing. Such an advanced DNA extraction system can be utilized in various fields including clinical analysis, pathogen detection, forensic analysis, and on-site detection.


Cardiac toxicity screening by echocardiography in healthy volunteers: a study of the effects of diurnal variation and use of a core laboratory on the reproducibility of left ventricular function measurement.
In investigational medicinal products testing centers (IMP), reliable methods for monitoring early signs of cardiotoxicity of a potential new drug in healthy volunteers are essential. This study examines what levels of left ventricular ejection fraction (LVEF) variance can be achieved with two-dimensional echocardiography (2DE) in a core laboratory versus a site laboratory. Diurnal variability of LVEF and diastolic parameters were also reviewed. 64 healthy males, (age range 18-40 years), with optimal echo windows were recruited. Two-dimensional and tissue Doppler (TDI) echocardiography was performed by one dedicated sonographer using an Acuson Sequoia C256 machine. Heart rate and blood pressure were recorded simultaneously. Echocardiograms were performed at set time points (0, 1, 4, and 20 hours) on all subjects. The images were analyzed independently by one on-site, unblinded, sonographer reader (site lab) and one experienced off-site blinded physician over reader (core lab). The core lab showed significantly less variance in LVEF measurements than the site lab (5.5% vs. 19.9%). There was no significant diurnal variation in mean blood pressure, LVEF or E:A ratio measurements over 20 hours. The core lab had better reproducibility and significantly less variance in LVEF measurements by 2DE than the site lab. There was no diurnal variation in LV function measurement.


Destination Memory in Korsakoff's Syndrome.
Context memory, or the ability to remember the context in which an episodic event has occurred (e.g., where and when an event took place), has been found to be compromised in Korsakoff's syndrome. This study examined whether a similar deficit would be observed for destination memory, that is, the ability to remember to whom an information was previously transmitted. Patients with Korsakoff's syndrome and healthy controls were instructed to tell proverbs to pictures of celebrities. In a subsequent recognition test, they had to indicate to which celebrity they had previously told the proverbs. Participants also completed a neuropsychological battery including a binding task in which they were required to associate letters with their correspondent locations to assess context memory. Results showed worse binding and destination memory in patients with Korsakoff's syndrome than in controls. In the Korsakoff group, destination memory was significantly correlated with and predicted by performances on the binding task. The binding process seems to be impaired in Korsakoff's syndrome, a deficit that may account for the destination memory compromise in the syndrome, and probably, for the difficulty to retrieve the "where and when" of an encountered event.


Current treatment options and strategies for myelodysplastic syndromes.
The myelodysplastic syndromes (MDS) are a group of heterogeneous disorders characterized by ineffective hematopoiesis. A number of scoring systems have been developed but they are of limited value in guiding individual patient treatment decisions. We have presented the benefits and limitations of various treatment approaches for MDS to help provide guidance in treatment decisions. An extensive literature search on PubMed from 2000 to 2008 was done using MESH keywords MDS, treatment, 5-azacytidine, decitabine, lenalidomide, antithymocyte globulin. In recent years there has been progress in supportive care and three agents have been approved for the treatment of MDS. With the exception of lenalidomide, which produced remarkable responses with 5q chromosomal deletion, these therapies benefit a minority of patients and the overall outcomes are still unsatisfactory. Allogeneic stem cell transplant remains the only curative option but the majority of MDS patients are not eligible because of older age and other medical problems. Immunosuppressive therapy is helpful in a fraction of patients. Several newer agents are being tested, offering promise for the future.


Hordenine, a single compound produced during barley germination, inhibits melanogenesis in human melanocytes.
Melanin plays an important role protecting skin against ultraviolet light injury. However, increased production and accumulation of melanin results in a large number of skin disorders. Here, we identified hordenine as an active compound from germinated barley (Hordeum vulgare L.) and investigated the effects of hordenine on melanogenesis and its mechanisms of action in human epidermal melanocytes. We measured melanin content, tyrosinase activity, expression of melanogenesis-related proteins, and cAMP production. Melanin content was significantly inhibited by hordenine. The intracellular cAMP level was also reduced by hordenine. In addition, expression of microphthalmia-associated transcription factor (MITF), an upstream transcription factor of tyrosinase as well as tyrosinase, tyrosinase related protein (TRP)-1, and TRP-2, was inhibited by hordenine. Taken together, these results show that hordenine inhibited melanogenesis by suppressing cAMP production, which is involved in the expression of melanogenesis-related proteins and suggest that hordenine may be an effective inhibitor of hyperpigmentation.


Reanalysis with optimized power of red blood cell acetylcholinesterase activity from a 1-year dietary treatment of dogs to chlorpyrifos.
A no-observed-effect level (NOEL) of 0.1 mg/kg/day was reported for inhibition of red blood cell (RBC) acetylcholinesterase (AChE) in two groups of Beagle dogs fed chlorpyrifos (0, 0.01, 0.03, 0.1, 1 or 3 mg/kg/day) in the diet for 1 or 2 years (McCollister et al., Food Cosmet. Toxicol. 12 (1974) 45-61). The statistical analyses were by t-test that had low statistical power due to small sample sizes. Common time points for blood samples in both phases allowed a reanalysis of the grouped data over a 1-year time period. The reanalysis increased statistical power by increasing the sample size to n=14 from n=3 or 4, and decreasing the variance, by statistical step-by-step aggregation of the data from both phases, both sexes, and four sample periods. Factors retained in the ANOVA were dose, sex, and phase (sex-by-dose was not significant). Contrasts with one-sided t-tests indicated the 1 and 3 mg/kg/day groups had significantly inhibited RBC AChE (P<0.0001). At alpha=0.05, the uncorrected one-sided model had 80% power to detect a 12% decrease, 93% power for a 15% decrease, and 99.5% power for a 20% decrease in AChE activity. Overall, the reanalysis had high power to detect a clinically significant decrease in RBC AChE activity, and substantiated the original NOEL for chronic treatment of dogs to dietary chlorpyrifos at 0.1 mg/kg/day.


Localization of c-mos mRNA around the animal pole in the zebrafish oocyte with Zor-1/Zorba.
In oocytes, many maternally supplied products are stored, and these products play important roles in cell cycle regulation and early development. Mos protein, which is coded on the c-mos gene, promotes oocyte maturation and is involved in MAP-kinase signaling pathway. In Xenopus, maternally supplied c-mos mRNA undergoes poly(A) addition, and translational activation via CPE (cytoplasmic polyadenylation element) and CPEB (CPE binding protein). The elongated poly(A) is shortened and the c-mos mRNA is degraded during early embryogenesis via EDEN (embryo deadenylation element) and EDEN-BP (EDEN-binding protein). We cloned the full-length zebrafish c-mos gene, which is conserved at the protein coding region in vertebrates. c-mos mRNA has two putative CPE sequences in its 3'UTR, which binds to zebrafish CPEB homologous protein, Zor-1. We could not observe EDEN sequence, and could not detect interaction between c-mos mRNA and zebrafish EDEN-BP homologous protein, Brul, even though immuno precipitation and RT-PCR experiments suggested that c-mos mRNA interacts with Zor-1 in vivo. Interestingly, we found c-mos mRNA is located in the animal cortex of zebrafish oocyte, where Zor-1 protein exists. Taken together, these results suggest that the animal cortex is the central core of oocyte maturation in zebrafish.


Analysis by RNA-RNA hybridization assay of intertypic rotaviruses suggests that gene reassortment occurs in vivo.
Antigenic characterization of human and animal rotaviruses by the plaque reduction neutralization assay has shown the existence of naturally occurring intertypes. Antiserum to M37, a rotavirus strain isolated from an asymptomatic neonate, neutralizes both Wa and ST3 strains, which are classified as serotype 1 and serotype 4 human rotaviruses, respectively. Likewise, antiserum to SB-1A, a porcine rotavirus, neutralizes rotavirus strains belonging to serotype 4 or 5. Plaque reduction neutralization assay of reassortant rotaviruses produced in vitro from these intertypes indicates that these viruses share one antigenically related outer capsid protein, VP3, with one serotype and another antigenically related outer capsid protein, VP7, with the other serotype. Thus, M37 is related to ST3 on the basis of its fourth-gene product, VP3, and to Wa on the basis of its ninth-gene product, VP7, whereas SB-1A is related to Gottfried (serotype 4 porcine rotavirus) via VP7 and to OSU (serotype 5 porcine rotavirus) via VP3. RNA-RNA hybridization studies revealed a high degree of homology between the VP3 or VP7 gene segments responsible for shared serotype specificity. Thus, the fourth gene segments of M37 and ST3 were highly homologous, while M37 and Wa had homology between their ninth gene segments. SB-1A and Gottfried were homologous not only with respect to the ninth gene but had complete homology in all other genes except the fourth gene. The fourth gene of SB-1A was highly homologous with the fourth gene of OSU. These observations suggested that SB-1A was a naturally occurring reassortant between Gottfried-like and OSU-like porcine rotavirus strains. Our observations also suggested that intertypes may result from genetic reassortment in nature.


Many-impurity scattering on the surface of a topological insulator.
We theoretically address the impact of a random distribution of non-magnetic impurities on the electron states formed at the surface of a topological insulator. The interaction of electrons with the impurities is accounted for by a separable pseudo-potential method that allows us to obtain closed expressions for the density of states. Spectral properties of surface states are assessed by means of the Green's function averaged over disorder realisations. For comparison purposes, the configurationally averaged Green's function is calculated by means of two different self-consistent methods, namely the self-consistent Born approximation (SCBA) and the coherent potential approximation (CPA). The latter is often regarded as the best single-site theory for the study of the spectral properties of disordered systems. However, although a large number of works employ the SCBA for the analysis of many-impurity scattering on the surface of a topological insulator, CPA studies of the same problem are scarce in the literature. In this work, we find that the SCBA overestimates the impact of the random distribution of impurities on the spectral properties of surface states compared to the CPA predictions. The difference is more pronounced when increasing the magnitude of the disorder.


Chemometrics in metabolomics--a review in human disease diagnosis.
Metabolomics is a post genomic research field concerned with developing methods for analysis of low molecular weight compounds in biological systems, such as cells, organs or organisms. Analyzing metabolic differences between unperturbed and perturbed systems, such as healthy volunteers and patients with a disease, can lead to insights into the underlying pathology. In metabolomics analysis, large amounts of data are routinely produced in order to characterize samples. The use of multivariate data analysis techniques and chemometrics is a commonly used strategy for obtaining reliable results. Metabolomics have been applied in different fields such as disease diagnosis, toxicology, plant science and pharmaceutical and environmental research. In this review we take a closer look at the chemometric methods used and the available results within the field of disease diagnosis. We will first present some current strategies for performing metabolomics studies, especially regarding disease diagnosis. The main focus will be on data analysis strategies and validation of multivariate models, since there are many pitfalls in this regard. Further, we highlight the most interesting metabolomics publications and discuss these in detail; additional studies are mentioned as a reference for the interested reader. A general trend is an increased focus on biological interpretation rather than merely the ability to classify samples. In the conclusions, the general trends and some recommendations for improving metabolomics data analysis are provided.


The expression of SMN1, MART3, GLE1 and FUS genes in spinal muscular atrophy.
Spinal muscular atrophy (SMA) is one of the most common genetic causes of death in infants due to a mutation of the motor neuron 1 (SMN1) gene. The SMN1 gene encodes for the multifunctional SMN protein. SMN has been shown to be implicated in pre-mRNA splicing, mRNA transport and translational control. Also other mRNA processing proteins, such as GLE1, Marten (MART3) and Fused in Sarcoma (FUS), have been linked to neurodegenerative diseases. The aim of the study was to determine the expression of SMN, GLE1, MART3 and FUS genes in cell lines of the fibroblasts derived from SMA patients and normal controls. Total RNA was extracted from purchased fibroblasts acquired from three SMA type I patients and fibroblasts of three age-matched healthy controls. The RNA was then subjected to qPCR analysis using primers specific for the GLE1, MART3, FUS and SMN1 genes vs. GAPDH as internal control gene. SMN1 mRNA levels were at least ×10 lower in fibroblasts of SMA patients compared to controls. Gle1 and MART3 gene expression was ×2 downregulated whereas FUS mRNA levels appeared to be ×3 upregulated in SMA cells when compared to controls. We found a high correlation between FUS gene expression level to the SMN1 at gene expression level of fibroblast cell lines of SMA type I patients (r = 0.994, p < 0.0001). Our preliminary data show an intriguing expression profile of Gle1, MART3 and FUS genes in SMA, and suggest a critical role of FUS protein in the SMA pathogenesis.


Olfactory learning-induced long-lasting enhancement of descending and ascending synaptic transmission to the piriform cortex.
Learning of a particularly difficult olfactory-discrimination (OD) task results in acquisition of rule learning. This remarkable enhancement in learning capability is accompanied by long-term enhancement of synaptic connectivity between piriform cortex (PC) pyramidal neurons. Because successful performance in the OD task requires integration of information about the identity and also about the reward value of odors, it is likely that a higher-order brain area would also be involved in rule learning acquisition and maintenance. The anterior PC (APC) receives a strong ascending input from the olfactory bulb, carrying information regarding olfactory cues in the environment. It also receives substantial descending input from the orbitofrontal cortex (OFC), which is thought to play an important role in encoding the predictive value of odor stimuli. Using in vivo recordings of evoked field postsynaptic potentials, we characterized the physiological properties of projections to APC from the OFC and examined whether descending and ascending synaptic inputs to the piriform cortex are modified after OD learning. We show that enhanced learning capability is accompanied by long-term enhancement of synaptic transmission in both the descending and ascending inputs. Long-term synaptic enhancement is not accompanied by modifications in paired-pulse facilitation, indicating that such modifications are likely postsynaptic. Predisposition for long-term potentiation induction was affected by previous learning, and surprisingly also by previous exposure to the odors and training apparatus. These data suggest that enhanced connectivity between the APC and its input sources is required for OD rule learning.


Arylneuraminidase activity of Pseudomonas aeruginosa does not degrade natural substrates such as human respiratory mucins.
The culture supernatant from a single Pseudomonas aeruginosa strain has been reported to show neuraminidase activity, leading to the speculation that this bacterium may use this enzyme as a virulence factor to act on host macromolecules. In order to extend this finding, we have examined the activity of concentrated P. aeruginosa culture supernatants and cells on synthetic and natural substrates containing sialic acid, such as human respiratory mucins. Four P. aeruginosa strains showed some activity on the synthetic substrate 4-methylumbelliferyl-alpha-D-N-acetylneuraminic acid but failed to liberate N-acetylneuraminic acid from six different natural substrates. Attempts to induce enzyme production by use of human respiratory mucins in the culture medium were also unsuccessful. The supernatants also showed N-acetyl-beta-D-glucosaminidase-like activity on a synthetic substrate but did not liberate N-acetylhexosamines from natural substrates. We conclude that the neuraminidase-like activity observed in P. aeruginosa can be defined as an arylneuraminidase and that the possession of a neuraminidase active on natural substrates is not a common attribute of P. aeruginosa strains.


Cryo-EM Structure and Molecular Dynamics Analysis of the Fluoroquinolone Resistant Mutant of the AcrB Transporter from Salmonella.
Salmonella is an important genus of Gram-negative pathogens, treatment of which has become problematic due to increases in antimicrobial resistance. This is partly attributable to the overexpression of tripartite efflux pumps, particularly the constitutively expressed AcrAB-TolC. Despite its clinical importance, the structure of the Salmonella AcrB transporter remained unknown to-date, with much of our structural understanding coming from the Escherichia coli orthologue. Here, by taking advantage of the styrene maleic acid (SMA) technology to isolate membrane proteins with closely associated lipids, we report the very first experimental structure of Salmonella AcrB transporter. Furthermore, this novel structure provides additional insight into mechanisms of drug efflux as it bears the mutation (G288D), originating from a clinical isolate of Salmonella Typhimurium presenting an increased resistance to fluoroquinolones. Experimental data are complemented by state-of-the-art molecular dynamics (MD) simulations on both the wild type and G288D variant of Salmonella AcrB. Together, these reveal several important differences with respect to the E. coli protein, providing insights into the role of the G288D mutation in increasing drug efflux and extending our understanding of the mechanisms underlying antibiotic resistance.


A transcriptomics-based drug repositioning approach to identify drugs with similar activities for the treatment of muscle pathologies in spinal muscular atrophy (SMA) models.
Spinal muscular atrophy (SMA) is a genetic neuromuscular disorder caused by the reduction of survival of motor neuron (SMN) protein levels. Although three SMN-augmentation therapies are clinically approved that significantly slow down disease progression, they are unfortunately not cures. Thus, complementary SMN-independent therapies that can target key SMA pathologies and that can support the clinically approved SMN-dependent drugs are the forefront of therapeutic development. We have previously demonstrated that prednisolone, a synthetic glucocorticoid (GC) improved muscle health and survival in severe Smn-/-;SMN2 and intermediate Smn2B/- SMA mice. However, long-term administration of prednisolone can promote myopathy. We thus wanted to identify genes and pathways targeted by prednisolone in skeletal muscle to discover clinically approved drugs that are predicted to emulate prednisolone's activities. Using an RNA-sequencing, bioinformatics, and drug repositioning pipeline on skeletal muscle from symptomatic prednisolone-treated and untreated Smn-/-; SMN2 SMA and Smn+/-; SMN2 healthy mice, we identified molecular targets linked to prednisolone's ameliorative effects and a list of 580 drug candidates with similar predicted activities. Two of these candidates, metformin and oxandrolone, were further investigated in SMA cellular and animal models, which highlighted that these compounds do not have the same ameliorative effects on SMA phenotypes as prednisolone; however, a number of other important drug targets remain. Overall, our work further supports the usefulness of prednisolone's potential as a second-generation therapy for SMA, identifies a list of potential SMA drug treatments and highlights improvements for future transcriptomic-based drug repositioning studies in SMA.


Potential approaches using teneligliptin for the treatment of type 2 diabetes mellitus: current status and future prospects.
Based on pharmacological properties and results from clinical studies, teneligliptin has a great potential to be used as an alternate-day therapy and also the daily dose can be reduced to 10 mg. Clinical data also suggest its excellent efficacy and safety among older subjects. We have reviewed and discussed potential approaches using teneligliptin for the treatment of type 2 diabetes mellitus (T2DM) including alternate-day therapy and reduction of dose from 20 mg to 10 mg per day. We have also discussed the potential of teneligliptin to address the needs of older patients with T2DM. It is an excellent option for use in older patients as studies in the geriatric population have shown encouraging results. Teneligliptin has a desirable pharmacokinetic profile that makes it a potential drug for use on an alternate-day basis. Teneligliptin has shown anti-diabetic efficacy even at a dose of 10 mg. These approaches may improve treatment satisfaction and patient compliance and can lower the cost; however, it is crucial to identify the subset of T2DM patients who can obtain maximum benefits. To verify these effects, large clinical investigations need to be planned and robust clinical evidence should be generated.


[Simultaneous operations in patients with cholelithiasis and gastroesophageal reflux disease].
The combination of the cholecystitis with gastro-esophageal reflux disease is registries about 12% of cases. On the basis of the CSRI of gastroenterology we underwent 36 patients with cholecystitis combined with gastro-esophageal reflux disease simultaneous surgeries. An analysis of the operation results has shown effectiveness and safety of using the methods by laparoscopic approach.