Emulsion pcr (Epcr) as a tool to improve the power of dgge analysis for microbial population studies

To the authors\u2019 knowledge, this is the first report of the use of emulsion-Polymerase chain reaction (e-PCR) coupled with denaturing gradient gel electrophoresis (DGGE) analysis. In the present work the effectiveness of ePCR in improving the power of the DGGE technique for microbial population studies was tested. Our results indicated that ePCR results in uniform amplification of several DNA molecules, overcoming the major limitations of conventional PCR, such as preferential amplification and DNA concentration dependence. Moreover, ePCR-DGGE resulted in higher sensitivity when compared to conventional PCR-DGGE methods used for studying microbial populations in a complex matrix. In fact, compared to conventional PCR, the DGGE profiles of ePCR products permitted the detection of a higher number of the species that were present in the tested sample

Characterization of microclimate and turbulent fluxes at a Mediterranean kiwi orchard covered with hail-protection net

Screens and covers are increasingly used to protect crops from pests and extreme meteorological events. Their use affects plant microclimate and physiological responses as well, but this is only partly understood, particularly when considering the interaction among the cover and the training system. This study focuses on the microclimatic effects of the use of a hail protection net and an horizontal (pergola) kiwifruit canopy. The system splits the orchard environment in three distinct layers, determining a shaded understory, a space comprised between the canopy and the net, and the atmosphere above the net. To accent the effects, we considered a high-water demanding crop-kiwifruit, in an environment characterized by high evaporative demand (Bernalda, southern Italy).Three full eddy covariance and radiative balance equipment were used to assess fluxes (carbon dioxide, water vapor, and sensible heat) in the three layers and monitor meteorological variables (air temperature and relative humidity, wind, upward and downward short and long wave radiation, carbon dioxide and water vapor concentrations). Data from a typical clear-sky summer day are considered in this paper.While the net strongly reduced wind speed, it had a modest impact on all other variables. Conversely, the tick canopy layer had a major impact on all variables, determining a highly shaded, cooler and more humid understory, with very light wind. Nevertheless, the combination of high relative humidity and presence of the net was able to reduce the net loss of longwave radiation from the canopy during night, mitigating its cooling under these conditions.The reduction in wind speed and the increase in incoming longwave radiation around the crop, observed at night time, indicate potential valuable mechanisms that may be exploited to decrease water needs and prevent late frosts in the context of climate change, where extreme climatic events are more frequent and crop water requirements continue to increase

Alcohol marketing in the Americas and Spain during the 2014 FIFA World Cup Tournament

Background and aims To identify the nature of visual alcohol references in alcohol advertisements during televised broadcasts of the 2014 FIFA World Cup Tournament matches and to evaluate cross-national differences according to alcohol marketing policy restrictiveness. Design Content analysis using the Delphi method and identification of in-game sponsorships. Setting Television broadcasts of the 2014 FIFA World Cup in Argentina, Brazil, Canada, Finland, France, Mexico, Spain and the United States. Cases Eighty-seven alcohol advertisements20 matches. Measurements Quantitative rating scales, combined with the Delphi rating technique, were used to determine compliance of the alcohol advertisements with the International Alliance for Responsible Drinking's (IARD) Guiding Principles. Recordings of five matches from four countries were also used to identify the number of in-and out-of-game alcohol brand appearances. Findings A total of 86.2% of all unique alcohol advertisements contained at least one violation of IARD's Guiding Principles, with violation rates ranging from 72.7% (Mexico) to 100% (Brazil). Countries with the least restrictive marketing policies had a higher prevalence of violations in guidelines designed to protect minors. There were 2.76 in-game alcohol brand appearances and 0.83 out-of-game alcohol brand appearances per minute. Brand appearances did not differ across countries or according to a country's marketing policy restrictiveness. Conclusions Self-regulation and statutory policies were ineffective at limiting alcohol advertising during the 2014 FIFA World Cup Tournament television broadcasts. Most advertisements contained content that violated the self-regulation codes, and there were high levels of within broadcast brand appearances.Pan American Health OrganizationAlcohol Research UKInstitute of Alcohol StudiesUniv Connecticut, Sch Med, Dept Community Med & Hlth Care, Farmington, CT USAUniv Fed Sao Paulo, Dept Psychiat, Sao Paulo, BrazilPan Amer Hlth Org, Alcohol & Subst Abuse, Washington, DC USADepartment of Psychiatry, Universidade Federal de São Paulo (UNIFESP), São Paulo, BrazilPAHO: SC-14-02239Web of Scienc

Effect of short-term hazelnut consumption on DNA damage and oxidized LDL in children and adolescents with primary hyperlipidemia : a randomized controlled trial

Children with primary hyperlipidemia are prone to develop premature atherosclerosis, possibly associated with increased oxidative stress. Nutritional therapy is the primary strategy in the treatment of hyperlipidemia and associated conditions. Dietary interventions with bioactive-rich foods, such as nuts, may contribute to the modulation of both lipid profile and the oxidative/antioxidant status. Our study aimed to assess the impact of a dietary intervention with hazelnuts on selected oxidative stress markers in children and adolescents with primary hyperlipidemia. A single-blind, 8-week, randomized, controlled, three-arm, parallel-group study was performed. Children and adolescents diagnosed with primary hyperlipidemia (n=60) received dietary guidelines and were randomized into three groups: group 1 received hazelnuts with skin (HZN+S), and group 2 hazelnuts without skin (HZN-S), at equivalent doses (15-30 g/day, based on body weight); group 3 (controls) received only dietary recommendations (no nuts). At baseline and after 8 weeks, plasma oxidized low-density lipoprotein (ox-LDL) concentrations, oxidative levels of DNA damage in PBMCs and potential correlation with changes in serum lipids were examined. A reduction of endogenous DNA damage by 18.9%\ub151.3% (P=.002) and 23.1%\ub147.9% (P=.007) was observed after HZN+S and HZN-S, respectively. Oxidatively induced DNA strand breaks decreased by 16.0%\ub138.2% (P=.02) following HZN+S treatment. Ox-LDL levels did not change after HZN+S intervention but positively correlated with total cholesterol and LDL cholesterol. A short-term hazelnut intervention improves cell DNA protection and resistance against oxidative stress but not ox-LDL in hyperlipidemic pediatric patients. The trial was registered at ISRCTN.com, ID no. ISRCTN12261900

Targeting Beta-Blocker Drug–Drug Interactions with Fibrinogen Blood Plasma Protein: A Computational and Experimental Study

In this work, one of the most prevalent polypharmacology drug–drug interaction events that occurs between two widely used beta-blocker drugs—i.e., acebutolol and propranolol—with the most abundant blood plasma fibrinogen protein was evaluated. Towards that end, molecular docking and Density Functional Theory (DFT) calculations were used as complementary tools. A fibrinogen crystallographic validation for the three best ranked binding-sites shows 100% of conformationally favored residues with total absence of restricted flexibility. From those three sites, results on both the binding-site druggability and ligand transport analysis-based free energy trajectories pointed out the most preferred biophysical environment site for drug–drug interactions. Furthermore, the total affinity for the stabilization of the drug–drug complexes was mostly influenced by steric energy contributions, based mainly on multiple hydrophobic contacts with critical residues (THR22: P and SER50: Q) in such best-ranked site. Additionally, the DFT calculations revealed that the beta-blocker drug–drug complexes have a spontaneous thermodynamic stabilization following the same affinity order obtained in the docking simulations, without covalent-bond formation between both interacting beta-blockers in the best-ranked site. Lastly, experimental ultrasound density and velocity measurements were performed and allowed us to validate and corroborate the computational obtained resultsThis research was funded by FCT/MCTES through national funds (Michael González-Durruthy, Riccardo Concu, and M. Natália D.S. Cordeiro), grant UID/QUI/50006/2020, as well as by Xunta de Galicia (Juan M. Ruso), grant ED41E2018/08S

Avaliação da presença de anticorpos IgG anti-Schistosoma mansoni no soro de pacientes com esquistossomose mansônica crônica, antes e após tratamento específico

The circumoval precipitin test (COPT), enzyme-linked immunosorbent assay (ELISA) and the immunoblotting anti-adult worm antigen (AWA) and soluble egg antigen (SEA) tests were applied to 17 chronically schistosome-infected patients for the detection of anti-Schistosoma mansoni antibodies before and on four occasions after oxamniquine administration over a period of six months. Compared to a control group, schistosomiasis patients showed high levels of IgG antibodies in AWA and SEA-ELISA. A decrease in IgG levels was observed six months after treatment, although negative reactions were not obtained. Significant decreases in IgG1, IgG3 and, mainly, IgG4, but not anti-SEA IgG2 levels were observed six months after treatment, again without negativity. Analysis of anti-AWA IgG antibodies by immunoblotting before treatment showed a 31 kDa strand in 14 patients (82%) which disappeared in three cases up to six months after treatment; furthermore, anti-SEA IgG antibodies showed the same band in nine patients (53%) before treatment, which disappeared in only four cases up to six months after treatment.Em 17 pacientes com infecção crônica por Schistosoma mansoni utilizaram-se os testes de reação periovular, imunoenzimático (ELISA) e imunoblotting, empregando-se antígenos obtidos a partir de vermes adultos (AWA) ou de ovos de S. mansoni (SEA), para detecção de anticorpos anti-S. mansoni, antes e em quatro ocasiões após tratamento com oxamniquine. Quando cotejados a grupo controle os pacientes esquistossomóticos revelaram altos níveis séricos de anticorpos IgG nos testes ELISA (anti-AWA e anti-SEA), não se observando, porém, negativação até seis meses após tratamento específico. Encontrou-se, entretanto, decréscimo significativo, sem negativação, dos níveis de IgG1, IgG3 e, principalmente, IgG4, quando se utilizou antígeno solúvel obtido a partir de ovos de S. mansoni (SEA), seis meses após administração de oxamniquine. O mesmo não foi observado no caso de anticorpos da subclasse IgG2. Nos imunoblottings efetuados com o emprego de antígeno de verme adulto (AWA), antes do tratamento com oxamniquine, evidenciou-se a presença de banda com 31 kDa em 14 (82%) dos 17 pacientes estudados, observando-se seu desaparecimento em três pacientes examinados seis meses após tratamento específico. Quando se utilizou antígeno obtido a partir de ovos de S. mansoni (SEA) a mesma banda foi evidenciada em nove pacientes, desaparecendo em quatro casos, após o tratamento

Anaplasma marginale infection in cattle from southwestern Amazonia.

study provides the first epidemiological data regarding infection by Anaplasma marginale in cattle reared in south-western Brazilian Amazonia. One simple procedure was adapted for the extraction of DNA from blood clots collected in seven microregions of Rondônia State and two mesoregions of Acre State. PCR method was used to asses the frequency of A. marginale infections in 4 to12-month-old cattle. The cattle infection was investigated by polymerase chain reaction (PCR) using the specific primer ?msp5? for A. marginale. The DNA amplifications revealed that the mean frequency of A. marginale infection was 98.6% (1,627/1,650) in samples from Rondonia, and 92.87% (208/225) in samples from Acre. The high frequency of A. marginale infections in 4 to 12- month-old cattle indicate a situation of enzootic stability in the studied areas and are comparable to those detected by immunodiagnosis in different endemic regions in Brazil. The DNA extraction of clotted blood method described here can be used for epidemiological studies on anaplasmosis and other bovine hemoparasites

Root production and quality attributes of sweet potato genotypes in Brasília - DF, Brazil, during two cropping seasons.

Avaliou-se nove genótipos de batata-doce: BRS Amélia, Beauregard, Brazlândia Branca, Brazlândia Rosada, Brazlândia Roxa, BRS Cuia, Princesa, BRS Rubissol e um genótipo local em delineamento de blocos casualizados, com quatro repetições. Estimou-se características relacionadas a produção de raízes, características de qualidade e danos causador por insetos