Diabetes induced immunological and biochemical changes in human colostrum

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental paracoccidioidomycosis in the mouse. III. Histopathological and immunological findings after intravenous infection in the presence or absence of previous immunization

Cinqüenta camundongos suíços, brancos, com quatro semanas de idade, foram inoculados com 5x10(5) formas leveduriformes, viáveis de Paracoccidiodes brasiliensis (cepa 18). Dez destes animais tinham sido previamente imunizados com antígeno particulado de P. brasiliensis, durante quatro semanas, por injeção intradérmica. Os controles consistiram de 10 animais que foram somente imunizados e 10 inoculados com solução salina estéril. Os animais foram sacrificados após 2, 4, 7, 11 e 16 semanas. Estudamos: 1) resposta de hipersensibilidade retardada medida pelo teste do coxim plantar, 24 horas antes do sacrifício; 2) anticorpo- gênese específica avaliada pelo teste de imunodifusão dupla em gel de ágar; 3) histopatologia dos pulmões, fígado, baço, supra-renal e rins. Observamos: 1) os animais imunizados desenvolveram resposta imunecelular mais intensa que os infectados; 2) a infecção deprimiu a resposta imunecelular dos animais imunizados; 3) a histopatologia da infecção endovenosa revelou inflamação granulomatosa sistêmica e progressiva. Os animais infectados após imunização prévia apresentaram inflamação pulmonar menos extensa, com granulomas menores e com reduzido número de fungos. O presente modelo murino de paracoccidioidomicose mimetiza alguns achados da forma humana subaguda da micose (doença sistêmica com depressão da imunidade celular). O esquema de imunização extrapulmonar utilizado foi capaz de induzir certo grau de proteção do pulmão contra um desafio infeccioso pelo P. brasiliensis.

Immunological evaluation of the intestinal mucosa of broiler chicks treated with Lactobacillus Spp. and challenged with Salmonella Enteritidis

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Comparison of parasitological, immunological and molecular methods for the diagnosis of leishmaniasis in dogs with different clinical signs

Aiming to improve the diagnosis of canine leishmaniasis (CanL) in an endemic area of the Northwest region of São Paulo State, Brazil, the efficacy of parasitological, immunological and molecular diagnostic methods were studied. Dogs with and without clinical sips of the disease and positive for Leishmania, by direct parasite identification on lymph node smears and/or specific antibody detection by ELISA, were selected for the study. According to the clinical signs, 89 dogs attending the Veterinary Hospital of UNESP in Aracatuba (SP, Brazil) were divided into three groups: symptomatic (36%), oligosymptomatic (22%) and asymptomatic (22%). Twenty-six dogs from an area non-endemic for CanL were used as negative controls (20%). Fine-needle aspiration biopsies (FNA) of popliteal lymph nodes were collected and Diff-Quick (R)-stained for optical microscopy. Direct immumofluorescence, immunocytochemistry and parasite DNA amplification by PCR were also performed. After euthanasia, fragments of popliteal lymph nodes, spleen, bone marrow and liver were collected and processed for HE and immunohistochemistry. Parasite detection by both HE and immunohistochemistry was specifically more effective in lymph nodes, when compared with the other organs. Immunolabeling provided higher sensitivity for parasite detection in the tissues. In the symptomatic group, assay sensitivity was 75.61% for direct parasite search on Diff-Quick (R)-stained FNAs, 92.68% for direct immunofluorescence, 92.68% for immunocytochemistry and 100% for PCR; the corresponding values in the other clinical groups were: 32, 60, 76 and 96% (oligosymptomatic), and 39.13, 73.91, 100 and 95.65% (asymptomatic). Results of the control animals from the CanL non-endemic area were all negative, indicating that the methods used were 100% specific. (C) 2006 Elsevier B.V. All rights reserved.

Effect of fluoride intake on carbohydrate metabolism, glucose tolerance, and insulin signaling

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Demographic and metabolic characteristics of individuals with progressive glucose tolerance

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dimorphism, thermal tolerance, virulence and Heat shock protein 70 transcription in different isolates of Paracoccidioides brasiliensis

The mycelia-to-yeast (M-Y) transition, thermal tolerance and virulence profiles were evaluated for nine isolates of Paracoccidioides brasiliensis, including samples from two of the three recently discovered cryptic species, as well as their relation to the partial sequence and transcription of the hsp70 gene. The isolates Bt84 and T10 (from PS2 species) took more time to convert to yeast form and presented elongated yeast cells at 36 degrees C. Arthroconidia production was also observed during the M-Y transition for some isolates. Our data confirm that the hsp70 transcription may be associated with thermal tolerance, but this does not seem to be directly related to high virulence profiles. The partial sequencing of this gene allowed the separation of our isolates into two clusters that correspond to the two sympatric cryptic species occurring in an area hyperendemic for PCM (Botucatu, SP, Brazil).

Drought tolerance is associated with rooting depth and stomatal control of water use in clones of Coffea canephora

center dot Background and Aims Drought is a major environmental constraint affecting growth and production of Coffea canephora. Selection of C. canephora clones has been largely empirical as little is known about how clones respond physiologically to drought. Using clones previously shown to differ in drought tolerance, this study aimed to identify the extent of variation of water use and the mechanisms responsible, particularly those associated morphological traits.center dot Methods Clones (14 and 120, drought-tolerant; 46 and 109A, drought-sensitive, based on their abilities to yield under drought) were grown in 120-L pots until they were 12-months old, when an irrigation and a drought treatment were applied; plants were droughted until the pressure potential (Psi(x)) before dawn (pre-dawn) reached -3.0 MPa. Throughout the drought period, Psi(x) and stomatal conductance (g(s)) were measured. At the end of the experiment, carbon isotope ratio and parameters from pressure-volume curves were estimated. Morphological traits were also assessed.center dot Key Results and Conclusions With irrigation, plant hydraulic conductance (K-L), midday Psi(x) and total biomass were all greater in clones 109A and 120 than in the other clones. Root mass to leaf area ratio was larger in clone 109A than in the others, whereas rooting depth was greater in drought-tolerant than in drought-sensitive clones. Predawn Psi(x) of -3.0 MPa was reached fastest by 109A, followed progressively by clones 46, 120 and 14. Decreases in g(s) with declining Psi(x), or increasing evaporative demand, were similar for clones 14, 46, and 120, but lower in 109A. Carbon isotope ratio increased under drought; however, it was lower in 109A than in other clones. For all clones, Psi(x), g(s) and KL recovered rapidly following re-watering. Differences in root depth, KL and stomatal control of water use, but not osmotic or elastic adjustments, largely explained the differences in relative tolerance to drought stress of clones 14 and 120 compared with clones 46 and 109A.

Overexpression of plant uncoupling mitochondrial protein in transgenic tobacco increases tolerance to oxidative stress

An Arabidopsis thaliana cDNA clone encoding a plant uncoupling mitochondrial protein (AtPUMP1) was overexpressed in transgenic tobacco plants. Analysis of the AtPUMP1 mRNA content in the transgenic lines, determined by Northern blot, revealed variable levels of transgene expression. Antibody probing of Western blots of mitochondrial proteins from three independent transgenic lines showed significant accumulation of AtPUMP1 in this organelle. Overproduction of AtPUMP1 in transgenic tobacco plants led to a significant increase in tolerance to oxidative stress promoted by exogenous hydrogen peroxide as compared to wild-type control plants. These results provide the first biological evidence for a role of PUMP in protection of plant cells against oxidative stress damage.

An Arabidopsis Mitochondrial Uncoupling Protein Confers Tolerance to Drought and Salt Stress in Transgenic Tobacco Plants

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Worm burden and immunological responses in Corriedale and Crioula Lanada sheep following natural infection with Haemonchus contortus

The effects of natural infection with Haemonchus contortus were compared in 15 Corriedale lambs and 18 Crioula Lanada lambs kept in the grazing conditions after weaning. The following parameters were assessed weekly for 9 consecutive weeks: number of eggs per gram of feces (EPG), packed cell volume (PCV), total serum proteins (TSP) and albumin (ALB). At 10 weeks, the animals were slaughtered and autopsied to evaluate to the worm burden and eosinophil, mast cell and globule leukocyte counts in histological sections of the abomasum. EPG counts and worm burden were significantly lower in Crioula lambs, which also showed higher levels of PCV, TSP and ALB, as well as higher eosinophil and globule leukocyte counts. Mast cell counts were similar for the two breeds. These results indicate that, under intensive pasture conditions, the Crioula Lanada breed shows a better response to natural infection with H. contortus, suggesting greater resistance to parasitism. (C) 2003 Elsevier B.V. All rights reserved.

Immunological responses and cytokine gene expression analysis to Cooperia punctata infections in resistant and susceptible Nelore cattle

Cellular and humoral immune response, as well as cytokine gene expression, was assessed in Nelore cattle with different degrees of resistance to Cooperia punctata natural infection. One hundred cattle (male, weaned, 11-12 months old), kept together on pasture, were evaluated. Faecal and blood samples were collected for parasitological and immunological assays. Based on nematode faecal egg counts (FEC) and worm burden, the seven most resistant and the eight most susceptible animals were selected. Tissue samples of the small intestine were collected for histological quantification of inflammatory cells and analysis of cytokine gene expression (IL-2, IL-4, IL-8, IL-1 2p35, IL-13, TNF-alpha, IFN-gamma, MCP-1, MCP-2, and MUC- 1) using real-time RT-PCR. Mucus samples were also collected for IgA levels determination. Serum IgG1 mean levels against C. punctata antigens were higher in the resistant group, but significant differences between groups were only observed 14 days after the beginning of the experiment against infective larvae (1-3) and 14 and 84 days against adult antigens. The resistant group also presented higher IgA levels against C. punctata (L3 and adult) antigens with significant difference 14 days after the beginning of the trial (P < 0.05). In the small-intestine mucosa, levels of IgA anti-L3 and anti-adult C. punctata were higher in the resistant group, compared with the susceptible group (P < 0.05). Gene expression of both T(H)2 cytokines (IL-4 and IL-13) in the resistant group and T(H)1 cytokines (IL-2, IL-1 2p35, IFN-gamma and MCP-1) in the susceptible group was up-regulated. Such results suggested that immune response to C. punctata was probably mediated by TH2 cytokines in the resistant group and by T(H)1 cytokines in the susceptible group. (C) 2008 Elsevier B.V. All rights reserved.

Aspirin increases tolerance to high temperature in seeds of Raphanus sativus L. cvar Early Scarlet Globe

The incubation of seeds of Raphanus sativus L. cvar Early Scarlet Globe with 10 mu M aspirin resulted in increase in the temperature range for germination. The analysis of percentage germination and germination rates indicated the increase in the optimum temperature from 21.4 to 26 degrees C although at 32.6 degrees C 80.8% of seeds germinated with aspirin and no germination in the control. The analysis of the kinetics of seed germination indicated that aspirin treatment resulted in germination by decreasing the enthalpy of activation of the process. The aspirin treatment also resulted in the synchronization of seed germination. on the base of our results we propose aspirin application in practice to increase the tolerance to high temperature and to synchronize seed germination at least in Raphanus sativus L. cvar early scarlet globe.