Substrato com Organosuper® para formação de mudas de pepineiro em ambientes protegidos e bandejas de poliestireno

Culture environments, trays and doses of organic compost were evaluated in the formation of cucumber seedlings (Cucumis sativus L.). Five environmental conditions were tested: (A1) a greenhouse with height of 2.5 m, covered with polyethylene film, (A2) nursery with height of 2.5 m, monofilament fabric, 50% shading, (A3) nursery with height of 2.5 m, heat-reflective screen, 50% shading, (A4) nursery with a height of 1.8 m, covered with coconut tree straw and (A5) greenhouse with height of 4.0 m, covered with polyethylene film, with zenith opening and thermo-reflective cloth under the plastic. Trays of 72 (R1) and 128 (R2) cells were filled with 93% soil and 7% organic compound (S1), 86% soil and 14% organic compound (S2) and 79% soil and 21% organic compound (S3). It was used a randomized design in split-split-plot scheme, with five replicates (environments x trays x substrates). The greenhouses provide the best environments for the formation of cucumber seedlings. A tray of 72 cells is the best container, promoting more vigorous seedlings in substrate with soil and 7 or 14% organic compound.Ambientes de cultivo, bandejas e doses de composto orgânico foram avaliados na formação de mudas de pepino (Cucumis sativus L.). Cinco ambientes de cultivo foram testados: (A1) estufa agrícola com altura de 2,5 m coberta com filme de polietileno; (A2) viveiro com altura de 2,5 m, tela de monofilamento com 50% de sombreamento; (A3) viveiro com altura de 2,5 m, tela termorrefletora, com 50% de sombreamento; (A4) viveiro com altura de 1,8 m, coberto com palha de coqueiro, e (A5) estufa agrícola com altura de 4,0 m, coberta com filme de polietileno, com abertura zenital e tela termorrefletora sob o filme. Bandejas de 72 (R1) e 128 (R2) células foram preenchidas com 93% de solo e 7% de composto orgânico (S1); 86% de solo e 14% de composto orgânico (S2), e 79% de solo e 21% de composto orgânico (S3). Utilizou-se um delineamento inteiramente casualizado, em esquema de parcelas subsubdivididas, com cinco repetições (ambientes x bandejas x substratos). As estufas agrícolas propiciam os melhores ambientes para a formação das mudas de pepino. A bandeja de 72 células é o melhor recipiente, promovendo plântulas mais vigorosas no substrato com solo e 7 ou 14 % de composto orgânico.22623

Association between circulating exhausted CD4+ T cells with poor meningococcal C conjugate vaccine antibody response in HIV-infected children and adolescents

OBJECTIVES: To investigate the expression levels of surface markers of activation (CD38 and HLA-DR), inhibition (PD-1, TIGIT and CD57) and co-stimulation (CD28 and CD127) on CD4+ T cells of children/adolescents with vertical HIV infection (HI patients) and HIV-uninfected (HU) controls vaccinated with the meningococcal C conjugate vaccine (MCC). METHODS: HI patients (n=12), aged 8–17 years, were immunized with two MCC injections, while HU controls (n=9), aged 5.3–10.7 years, received a single MCC dose (as per national recommendation at the time of this study, a single MCC vaccine dose should be given for healthy children and youth aged 1–18 years). The HI patients were categorized according to the combined antiretroviral therapy (cART) treatment. Blood samples were obtained before vaccination, after priming, and after the administration of a booster dose of vaccine to determine the serum bactericidal antibody (SBA) titers and the expression levels of surface markers on CD4+ T cells by flow cytometry. The levels of serum cytokines, IL-4 and CXCL-13 were also measured using Luminex kits. RESULTS: The co-expression of the TIGIT-HLA-DR-CD38 molecules increased in the CD4+ T cells of HI patients/ no-cART who also showed a lower frequency of CD127+CD28+ CD4+ T cells than HI patients/cART and HU group subjects. There were significant negative correlations between the frequency of exhausted CD4+ T cells and the SBA response. IL-4 levels were higher in HI patients/cART and positively correlated with SBA titers but negatively associated with the expression of exhaustion markers. Moreover, the CXCL-13 levels were positively correlated with the exhausted CD4+ T cells. CONCLUSION: The results of our study suggest that the co-expression of exhaustion markers and/or loss of co-stimulatory molecules influence the SBA response in HI patients

Detecção molecular de Escherichia coli enteropatogênica em psitacídeos assintomáticos em cativeiro

Psittaciformes are one of the most endangered groups of birds, and several Brazilian species are classified between vulnerable and critically endangered. It is thus necessary to identify agents that cause infections in captive wild animals and to assess the risks posed thereof and to design interventions to minimize the possibility of disease outbreaks, leading to the conservation of endangered species. The purpose of this study was to identify enteropathogenic Escherichia coli (EPEC) cloacal isolates from asymptomatic psittacines in captivity and evaluate the distribution of the EPEC pathotype. Cloacal swabs were obtained from 46 asymptomatic birds, and resulting isolates were tested by polymerase chain reaction (PCR) for the presence of the attaching and effacing gene (eae) and bundle-forming pilus structural gene (bfpA) of EPEC. Samples from several species were tested, and three samples were found to be positive for the eae and bfpA genes and characterized as typical EPEC. This is the first report of this pathotype in asymptomatic psittacines. Although certain E. coli strains are more pathogenic than others, various factors should be considered when determining the potential of E. coli isolates to cause disease in captive psittacines. Birds that are positive for the EPEC (typical) strain could be zoonotic sources of infection, and may have acquired these strains through contact with humans or domestic animals. These findings may also be valuable for the long-term management of endangered species ex situ as one EPEC sample was isolated from a Red-tailed Amazon (Amazona brasiliensis).Os psitacídeos são um dos grupos de aves mais ameaçadas no mundo e diversas espécies brasileiras são classificadas desde vulneráveis à criticamente ameaçadas de extinção. Torna-se, portanto, necessário identificar os agentes que causam infecções em animais selvagens em cativeiro e determinar os riscos relacionados de modo a intervir sobre os fatores envolvidos para diminuir a possibilidade de surtos de doenças e promover a conservação de espécies ameaçadas. O objetivo deste estudo foi identificar Escherichia coli Enteropatogência (EPEC) de isolados cloacais de psitacídeos assintomáticos em cativeiro e avaliar a distribuição do patotipo EPEC. Suabes cloacais foram coletados de 46 psitacídeos assintomáticos e os isolados foram testados pela reação em cadeia pela polimerase (PCR) para a presença do gene attaching and effacing (eae) e bundle forming pilus (bfpA) de EPEC. Amostras oriundas de diversas espécies foram testadas e três amostras resultaram positivas para os genes eae e bfp e caracterizadas como EPEC típicas. Esse é o primeiro relato em psitacídeos assintomáticos para esse patotipo. Apesar de que algumas cepas de E.coli serem mais patogênicas do que outras, diversos fatores devem ser considerados para determinar o potencial de isolados de E.coli de causar doença em psitacídeos em cativeiro. Aves positivas para cepas de EPEC (típicas) poderiam ser fontes de infecção zoonóticas e adquirir essas cepas através do contato com humanos e animais domésticos. Esses achados também podem ser valiosos para o manejo a longo prazo de espécies ameaçadas ex situ já que uma amostra de EPEC foi isolada de um Papagaio-de-cara-roxa (Amazona brasiliensis).Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) [2010/51015-0]Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP

Development of galactomannan membranes from seeds of Cassia grandis for immobilization of Caesalpinia ferrea pod lectin (Cfepl)

Galactomannans are polysaccharides present in the endosperm of numerous plants, particularly the Leguminosae, with several functions, including reserve carbohydrates. Polysaccharides membranes have been used as cross-linked matrix for immobilization of different biomolecules. Lectins are ubiquitous proteins in nature and can be used for a variety of biomedical applications. The aim of this work was the development of galactomannan membranes for immobilization of the Caesalpinia ferrea pod lectin (CfePL). The galactomannan from Cassia grandis seeds was obtained by aqueous extraction (20%), followed by salt precipitation (0.1M NaCl), centrifugation and washes, twice with ethanol (46% and 100%) and finally with acetone. The yield of the galactomannan extraction was determined by phenol sulfuric acid method. Different concentrations of galactomannan (0.5 1.5% w/v) and glycerol (0 0.3% v/v) were solubilized in distilled water under magnetic stirring (12h), placed on plates and then dried at 40ºC for 16h in order to obtain membranes. CfePL was obtained by saline extraction (10% NaCl), followed by affinity chromatography (chitin) and its immobilization was obtained by adding 0.5 mg/mL in galactomannan solution. The hemagglutinating activity was evaluated to confirm the activity of the immobilized lectin. The extraction yield for the galactomannan was approximately 97.33% and the most efficient membrane for lectin immobilization was obtained with 0.8% of galactomannan and 0.2% of glycerol. CfePL immobilization in galactomannan membranes was confirmed by hemagglutinating activity and by Fourier transform infrared spectroscopy (FTIR). These results suggest promising applications in medical therapy, especially on wound healing dressing of CfePL lectin immobilized on galactomannan membranes.info:eu-repo/semantics/publishedVersio

DNA methylation patterns in bladder cancer and washing cell sediments: a perspective for tumor recurrence detection

<p>Abstract</p> <p>Background</p> <p>Epigenetic alterations are a hallmark of human cancer. In this study, we aimed to investigate whether aberrant DNA methylation of cancer-associated genes is related to urinary bladder cancer recurrence.</p> <p>Methods</p> <p>A set of 4 genes, including <it>CDH1 </it>(E-cadherin), <it>SFN </it>(stratifin), <it>RARB </it>(retinoic acid receptor, beta) and <it>RASSF1A </it>(Ras association (RalGDS/AF-6) domain family 1), had their methylation patterns evaluated by MSP (Methylation-Specific Polymerase Chain Reaction) analysis in 49 fresh urinary bladder carcinoma tissues (including 14 cases paired with adjacent normal bladder epithelium, 3 squamous cell carcinomas and 2 adenocarcinomas) and 24 cell sediment samples from bladder washings of patients classified as cancer-free by cytological analysis (control group). A third set of samples included 39 archived tumor fragments and 23 matched washouts from 20 urinary bladder cancer patients in post-surgical monitoring. After genomic DNA isolation and sodium bisulfite modification, methylation patterns were determined and correlated with standard clinic-histopathological parameters.</p> <p>Results</p> <p><it>CDH1 </it>and <it>SFN </it>genes were methylated at high frequencies in bladder cancer as well as in paired normal adjacent tissue and exfoliated cells from cancer-free patients. Although no statistically significant differences were found between <it>RARB </it>and <it>RASSF1A </it>methylation and the clinical and histopathological parameters in bladder cancer, a sensitivity of 95% and a specificity of 71% were observed for <it>RARB </it>methylation (Fisher's Exact test (p < 0.0001; OR = 48.89) and, 58% and 17% (p < 0.05; OR = 0.29) for <it>RASSF1A </it>gene, respectively, in relation to the control group.</p> <p>Conclusion</p> <p>Indistinct DNA hypermethylation of <it>CDH1 </it>and <it>SFN </it>genes between tumoral and normal urinary bladder samples suggests that these epigenetic features are not suitable biomarkers for urinary bladder cancer. However, <it>RARB </it>and <it>RASSF1A </it>gene methylation appears to be an initial event in urinary bladder carcinogenesis and should be considered as defining a panel of differentially methylated genes in this neoplasia in order to maximize the diagnostic coverage of epigenetic markers, especially in studies aiming at early recurrence detection.</p