Optimización de la técnica SSCP-HD para la detección de polimorfismos en el exón 3 del gen ADIPOQ

Los polimorfismos del gen de la adiponectina están relacionados con alteraciones en su expresión con la subsecuente disminución en los niveles plasmáticos de adiponectina, esto último se encuentra asociado a la insulino-resistencia, obesidad e hipertensión arterial, todos factores de riesgo para enfermedades cardiovasculares y oncológicas. El objetivo del trabajo fue estandarizar una reacción de amplificación, PCR, seguida de detección mediante SSCPHD (polimorfismo de conformación de cadena simple-heterodúplex) para el análisis de los polimorfismos de simple nucleótido (SNPs) presentes en el exón 3 del gen ADIPOQ. Las extracciones de ADN se realizaron por el método Salting-Out modificado, los cebadores fueron analizados mediante programas bioinformáticos, y los fragmentos obtenidos fueron analizados mediante la técnica de Polimorfismo en la Cadena Simple-Heterodúplex (SSCP-HD). Con las condiciones utilizadas se logró la amplificación del exón 3 de ADIPOQ, identificándose patrones de corridas mediante la técnica de SSCP-HD que facilitarán el estudio posterior de los polimorfismos de simple nucleótido en el gen seleccionado.Polymorphisms of the adiponectin gene are associated with alterations in expression with subsequent decrease in plasma levels of adiponectin. The latter is associated with insulin resistance, obesity, and hypertension, all risk factors for cardiovascular disease and cancer. The objective of the present work was to standardize an amplification reaction, PCR, followed by detection by SSCPHD (strand conformation polymorphism-heteroduplex simple) for the analysis of single nucleotide polymorphisms (SNPs) present in exon 3 of the gene ADIPOQ. DNA extractions were performed by the modified salting-out method, the primers were analyzed using bioinformatics programs, and the fragments were analyzed by the technique of Polymorphism, Single-StrandedHeteroduplex (SSCP-HD). With the conditions used, the amplification of exon 3 of ADIPOQ was possible, identifying patterns of runs using the technique of SSCP-HD which will facilitate further studies of single nucleotide polymorphisms in the gene selected.Fil: González Gimenez, María del Carmen. Universidad Nacional de Misiones. Facultad de Cs.exactas Quimicas y Naturales. Departamento de Bioquimica Clinica. Laboratorio de Biotecnologia Molecular; ArgentinaFil: Tiscornia, María Mercedes. Universidad Nacional de Misiones. Facultad de Cs.exactas Quimicas y Naturales. Departamento de Bioquimica Clinica. Laboratorio de Biotecnologia Molecular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Hahn, Romina. Universidad Nacional de Misiones. Facultad de Cs.exactas Quimicas y Naturales. Departamento de Bioquimica Clinica. Laboratorio de Biotecnologia Molecular; ArgentinaFil: Zapata, Pedro Dario. Universidad Nacional de Misiones. Facultad de Cs.exactas Quimicas y Naturales. Departamento de Bioquimica Clinica. Laboratorio de Biotecnologia Molecular; ArgentinaFil: Bonneau, Graciela Alicia. Universidad Nacional de Misiones. Facultad de Ciencias Exactas, Químicas y Naturales; Argentin

Validation of the German version of the Southampton mindfulness questionnaire (SMQ)

Objectives: The aim of the study was to assess the convergent and divergent validity, reliability, utility, and treatment sensitivity of a newly translated German version of the Southampton Mindfulness Questionnaire (SMQ). The SMQ is a 16-item instrument measuring mindful awareness of distressing thoughts, images, and perceptions, developed originally within the mindfulness for psychosis field. Methods: Overall, three studies were conducted, comprising (1) a non-clinical sample of n = 848 (638 community sample and 210 meditators); (2) a clinical sample of n = 213 (106 schizophrenia and 107 depression); and (3) a clinical sample with n = 122 participants with emotional disorders within a randomized controlled study, of which 30 participants were also included in study 2. To assess convergent validity, participants completed the SMQ, Freiburg Mindfulness Inventory (FMI), and Comprehensive Inventory of Mindfulness Experiences (CHIME). To measure divergent validity, participants completed the Brief Symptom Inventory 18 (BSI-18), Positive and Negative Affect Schedule (PANAS), Brief Experiential Avoidance Questionnaires (BEAQ), and Anxiety Sensitivity Index 3 (ASI-3). Results: Mean internal consistency (α = 0.89) and convergent (r = 0.66 to 0.73) and divergent validity (r = − 0.09 to − 0.50) were established and sensitivity to change over time following treatment (d = 0.86) was shown. For the clinical sample, a single-factor structure is suggested by principal component analysis. Conclusions: Results provide first evidence for the utility of the German version of the SMQ for clinical practice and research in healthy individuals, meditators, and clinical groups. Further research is needed to examine the underlying construct of mindfulness

Clozapine modulates retinoid homeostasis in human brain and normalizes serum retinoic acid deficit in patients with schizophrenia

The atypical antipsychotic clozapine is one of the most potent drugs of its class, yet its precise mechanisms of action remain insufficiently understood. Recent evidence points toward the involvement of endogenous retinoic acid (RA) signaling in the pathophysiology of schizophrenia. Here we investigated whether clozapine may modulate RA-signaling. Effects of clozapine on the catabolism of all-trans RA (at-RA), the biologically most active metabolite of Vitamin A, were assessed in murine and human brain tissue and peripheral blood-derived mononuclear cells (PBMC). In patients with schizophrenia with and without clozapine treatment and matched healthy controls, at-RA serum levels and blood mRNA expression of retinoid-related genes in PBMCs were quantified. Clozapine and its metabolites potently inhibited RA catabolism at clinically relevant concentrations. In PBMC-derived microsomes, we found a large interindividual variability of the sensitivity toward the effects of clozapine. Furthermore, at-RA and retinol serum levels were significantly lower in patients with schizophrenia compared with matched healthy controls. Patients treated with clozapine exhibited significantly higher at-RA serum levels compared with patients treated with other antipsychotics, while retinol levels did not differ between treatment groups. Similarly, in patients without clozapine treatment, mRNA expression of RA-inducible targets CYP26A and STRA6, as well as at-RA/retinol ratio, were significantly reduced. In contrast, clozapine-treated patients did not differ from healthy controls in this regard. Our findings provide the first evidence for altered peripheral retinoid homeostasis in schizophrenia and suggest modulation of RA catabolism as a novel mechanism of action of clozapine, which may be useful in future antipsychotic drug development

A Two-Biomarker Model Predicts Mortality in the Critically Ill with Sepsis.

RATIONALE: Improving the prospective identification of patients with systemic inflammatory response syndrome (SIRS) and sepsis at low risk for organ dysfunction and death is a major clinical challenge. OBJECTIVES: To develop and validate a multibiomarker-based prediction model for 28-day mortality in critically ill patients with SIRS and sepsis. METHODS: A derivation cohort (n = 888) and internal test cohort (n = 278) were taken from a prospective study of critically ill intensive care unit (ICU) patients meeting two of four SIRS criteria at an academic medical center for whom plasma was obtained within 24 hours. The validation cohort (n = 759) was taken from a prospective cohort enrolled at another academic medical center ICU for whom plasma was obtained within 48 hours. We measured concentrations of angiopoietin-1, angiopoietin-2, IL-6, IL-8, soluble tumor necrosis factor receptor-1, soluble vascular cell adhesion molecule-1, granulocyte colony-stimulating factor, and soluble Fas. MEASUREMENTS AND MAIN RESULTS: We identified a two-biomarker model in the derivation cohort that predicted mortality (area under the receiver operator characteristic curve [AUC], 0.79; 95% confidence interval [CI], 0.74-0.83). It performed well in the internal test cohort (AUC, 0.75; 95% CI, 0.65-0.85) and the external validation cohort (AUC, 0.77; 95% CI, 0.72-0.83). We determined a model score threshold demonstrating high negative predictive value (0.95) for death. In addition to a low risk of death, patients below this threshold had shorter ICU length of stay, lower incidence of acute kidney injury, acute respiratory distress syndrome, and need for vasopressors. CONCLUSIONS: We have developed a simple, robust biomarker-based model that identifies patients with SIRS/sepsis at low risk for death and organ dysfunction

Long-term treatment with chaethomellic acid A reduces glomerulosclerosis and arteriolosclerosis in a rat model of chronic kidney disease

The high prevalence of end-stage renal disease emphasizes the failure to provide therapies to effectively prevent and/or reverse renal fibrosis. Therefore, the aim of this study was to evaluate the effect of long-term treatment with chaethomellic acid A (CAA), which selectively blocks Ha-Ras farnesylation, on renal mass reduction-induced renal fibrosis. Male Wistar rats were sham-operated (SO) or subjected to 5/6 renal mass reduction (RMR). One week after surgery, rats were placed in four experimental groups: SO:SO rats without treatment (n = 13); SO + CAA: SO rats treated with CAA (n = 13); RMR:RMR rats without treatment (n = 14); and RMR + CAA:RMR rats treated with CAA (n = 13). CAA was intraperitoneally administered in a dose of 0.23 μg/kg three times a week for six months. Renal fibrosis was evaluated by two-dimensional ultrasonography and histopathological analysis. The kidneys of the RMR animals treated with CAA showed a significantly decrease in the medullary echogenicity (p < 0.05) compared with the RMR rats that received no treatment. Glomerulosclerosis and arteriolosclerosis scores were significantly lower (p < 0.001) in the RMR + CAA group when compared with the RMR group. There were no significant differences in interstitial fibrosis, interstitial inflammation and tubular dilatation scores between the RMR + CAA and RMR groups. These data suggest that CAA can be a potential future drug to attenuate the progression of chronic kidney disease.This work is supported by : European Investment Funds by FEDER/COMPETE/POCI– Operacional Competitiveness and Internacionalization Programme, under Project POCI-01-0145-FEDER-006958 and National Funds by FCT - Portuguese Foundation for Science and Technology, under the project UID/AGR/04033/2013; and by European Investment Funds by FEDER/COMPETE/POCI– Operacional Competitiveness and Internacionalization Programme, under Project POCI-01-0145-FEDER-016728 and National Funds by FCT - Portuguese Foundation for Science and Technology, under the project PTDC/DTP-DES/6077/2014.info:eu-repo/semantics/publishedVersio

A tale of worldwide success: Behind the scenes of Carex (Cyperaceae) biogeography and diversification

The megadiverse genus Carex (c. 2000 species, Cyperaceae) has a nearly cosmopolitan distribution, displaying an inverted latitudinal richness gradient with higher species diversity in cold-temperate areas of the Northern Hemisphere. Despite great expansion in our knowledge of the phylogenetic history of the genus and many molecular studies focusing on the biogeography of particular groups during the last few decades, a global analysis of Carex biogeography and diversification is still lacking. For this purpose, we built the hitherto most comprehensive Carex-dated phylogeny based on three markers (ETS–ITS–matK), using a previous phylogenomic Hyb-Seq framework, and a sampling of two-thirds of its species and all recognized sections. Ancestral area reconstruction, biogeographic stochastic mapping, and diversification rate analyses were conducted to elucidate macroevolutionary biogeographic and diversification patterns. Our results reveal that Carex originated in the late Eocene in E Asia, where it probably remained until the synchronous diversification of its main subgeneric lineages during the late Oligocene. E Asia is supported as the cradle of Carex diversification, as well as a “museum” of extant species diversity. Subsequent “out-of-Asia” colonization patterns feature multiple asymmetric dispersals clustered toward present times among the Northern Hemisphere regions, with major regions acting both as source and sink (especially Asia and North America), as well as several independent colonization events of the Southern Hemisphere. We detected 13 notable diversification rate shifts during the last 10 My, including remarkable radiations in North America and New Zealand, which occurred concurrently with the late Neogene global cooling, which suggests that diversification involved the colonization of new areas and expansion into novel areas of niche space.This work was carried out with financial support by the National Science Foundation (Award #1255901 to ALH and Award #1256033 to EHR), the Spanish Ministry of Economy and Competitiveness (project CGL2016–77401‐P to SM-B and ML), the USDA National Institute of Food and Agriculture (McIntire Stennis project 1018692 to DS) as well as postdoctoral fellowships towards SM‐B (Universidad Pablo de Olavide, PP16/12‐APP), and PJ‐M (National Science Foundation, Award #1256033, and the Smithsonian Postdoctoral Fellowship program)

Measurement of σ(Λb)/σ(B0)×BR(Λb→Λcπ−)/BR(B0→D+π−)\sigma(\Lambda_b)/\sigma(B^0) \times BR(\Lambda_b\to\Lambda_c\pi^-) / BR(B^0\to D^+\pi^-) in ppˉp\bar{p} Collisions at s=1.96\sqrt{s}=1.96 TeV

We present the first observation of the baryon decay $\Lambda_b\to\Lambda_c\pi^-$ followed by $\Lambda_c\to p K^-\pi^+$ in 106 pb-1 of $p\bar{p}$ collisions at $\sqrt{s} = 1.96$ TeV in the CDF experiment. In order to reduce systematic error, the measured rate for $\Lambda_b$ decay is normalized to the kinematically similar meson decay $B^0\to D^+\pi^-$ followed by $D^+\to\pi^+K^-\pi^+$. We report the ratio of production cross sections ($\sigma$) times the ratio of branching fractions (BR) for the momentum region integrated above $p_T > 6$ GeV/c and pseudorapidity range $|\eta| < 1.3$: $\sigma(p\bar{p}\to \Lambda_b X) / \sigma (p\bar{p}\to B^0 X) \times BR(\Lambda_b\to\Lambda_c\pi^-) / BR(B^0\to D^+\pi^-) = 0.82 \pm 0.08(stat) \pm 0.11(syst) \pm 0.22 (BR(\Lambda_c\to p K^-\pi^+))$.Comment: Submitted to Phys.Rev.Let

Wired for motherhood: induction of maternal care but not maternal aggression in virgin female CD1 mice

Virgin adult female mice display nearly spontaneous maternal care towards foster pups after a short period of sensitization. This indicates that maternal care is triggered by sensory stimulation provided by the pups and that its onset is largely independent on the physiological events related to gestation, parturition and lactation. Conversely, the factors influencing maternal aggression are poorly understood. In this study, we sought to characterize two models of maternal sensitization in the outbred CD1 strain. To do so, a group of virgin females (godmothers) were exposed to continuous cohabitation with a lactating dam and their pups from the moment of parturition, whereas a second group (pup-sensitized females), were exposed 2 h daily to foster pups. Both groups were tested for maternal behavior on postnatal days 2-4. Godmothers expressed full maternal care from the first test. Also, they expressed higher levels of crouching than dams. Pup-sensitized females differed from dams in all measures of pup-directed behavior in the first test, and expressed full maternal care after two sessions of contact with pups. However, both protocols failed to induce maternal aggression toward a male intruder after full onset of pup-directed maternal behavior, even in the presence of pups. Our study confirms that adult female mice need a short sensitization period before the onset of maternal care. Further, it shows that pup-oriented and non-pup-oriented components of maternal behavior are under different physiological control. We conclude that the godmother model might be useful to study the physiological and neural bases of the maternal behavior repertoire

Effect of Acute Exposure to Moderate Altitude on Muscle Power: Hypobaric Hypoxia vs. Normobaric Hypoxia

When ascending to a higher altitude, changes in air density and oxygen levels affect the way in which explosive actions are executed. This study was designed to compare the effects of acute exposure to real or simulated moderate hypoxia on the dynamics of the force-velocity relationship observed in bench press exercise. Twenty-eight combat sports athletes were assigned to two groups and assessed on two separate occasions: G1 (n = 17) in conditions of normoxia (N1) and hypobaric hypoxia (HH) and G2 (n = 11) in conditions of normoxia (N2) and normobaric hypoxia (NH). Individual and complete force-velocity relationships in bench press were determined on each assessment day. For each exercise repetition, we obtained the mean and peak velocity and power shown by the athletes. Maximum power (Pmax) was recorded as the highest Pmean obtained across the complete force-velocity curve. Our findings indicate a significantly higher absolute load linked to Pmax (~3%) and maximal strength (1RM) (~6%) in G1 attributable to the climb to altitude (P<0.05). We also observed a stimulating effect of natural hypoxia on Pmean and Ppeak in the middle-high part of the curve (≥60 kg; P<0.01) and a 7.8% mean increase in barbell displacement velocity (P<0.001). No changes in any of the variables examined were observed in G2. According to these data, we can state that acute exposure to natural moderate altitude as opposed to simulated normobaric hypoxia leads to gains in 1RM, movement velocity and power during the execution of a force-velocity curve in bench press.This study has been supported by a Grant from the Ministry of education, culture and Sport of Spain, Reference 14/UPB10/07