Typing Candida albicans oral isolates from healthy Brazilian schoolchildren using multilocus enzyme electrophoresis reveals two highly polymorphic taxa

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

Development of an enzyme-linked immunosorbent assay for detection of IgM antibodies to Babesia bigemina in cattle

A crude antigenic preparation of Babesia bigemina was used to develop an ELISA for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Negative sera from cattle imported from tick-free areas, serum samples collected from infected B. bigemina cattle were used to validate the test. The specificity was 94% and sensitivity of the Elisa 87.5%. Sera from 385 cattle deriving from areas free from tick-borne diseases, which were submitted to a preimmunization process, were screened by this technique. The Elisa detected seroconversion on the 14th day post-inoculation in animals either infested with Boophilus microplus ticks (infected with B. bigemina), or inoculated with B. bigemina infected blood. Antibody titers decreased after day 33; however, all animals remained positive until the end of the experiment (124 days). The ELISA described may prove to be an appropriate serological test for the detection of IgM antibodies against B. bigemina.

Determinação de carbaril utilizando testes ELISA (Enzyme-linked immunosorbent assay) e CLAE com detecção por arranjo de diodos

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

Callus cell culture of Pothomorphe umbellata (L.) under stress condition leads to high content of peroxidase enzyme

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

Maternal protein restriction affects gene expression and enzyme activity of intestinal disaccharidases in adult rat offspring

This study investigated the consequences of intrauterine protein restriction on the gastrointestinal tract and particularly on the gene expression and activity of intestinal disaccharidases in the adult offspring. Wistar rat dams were fed isocaloric diets containing 6% protein (restricted, n = 8) or 17% protein (control, n = 8) throughout gestation. Male offspring (n = 5-8 in each group) were evaluated at 3 or 16 weeks of age. Maternal protein restriction during pregnancy produced offspring with growth restriction from birth (5.7 ± 0.1 vs 6.3 ± 0.1 g; mean ± SE) to weaning (42.4 ± 1.3 vs 49.1 ± 1.6 g), although at 16 weeks of age their body weight was similar to control (421.7 ± 8.9 and 428.5 ± 8.5 g). Maternal protein restriction also increased lactase activity in the proximal (0.23 ± 0.02 vs 0.15 ± 0.02), medial (0.30 ± 0.06 vs 0.14 ± 0.01) and distal (0.43 ± 0.07 vs 0.07 ± 0.02 U·g-1·min-1) small intestine, and mRNA lactase abundance in the proximal intestine (7.96 ± 1.11 vs 2.38 ± 0.47 relative units) of 3-week-old offspring rats. In addition, maternal protein restriction increased sucrase activity (1.20 ± 0.02 vs 0.91 ± 0.02 U·g-1·min-1) and sucrase mRNA abundance (4.48 ± 0.51 vs 1.95 ± 0.17 relative units) in the duodenum of 16-week-old rats. In conclusion, the present study shows for the first time that intrauterine protein restriction affects gene expression of intestinal enzymes in offspring.

Teste imunoenzimático (enzyme-linked immunosorbent assay) para diagnóstico da cisitcercose bovina e estudo da cinética de produção de anticorpos contra-Cysticercus bovis

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Enzyme activity and microbial biomass in an Oxisol amended with sewage sludge contaminated with nickel

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

Preparation of silica with controlled pore sizes for enzyme immobilization

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

Lipoperoxidation and cyclooxygenase enzyme inhibitory piperidine alkaloids from Cassia spectabilis green fruits

Phytochernical work in the search for bioactive metabolites from the methanolic extract of Senna spectabilis green fruits led to the isolation of a new piperidine alkaloid, (+)-3-O-feruloylcassine (1), in addition to the known (-)-spectaline (2) and (-)-3-O-acetylspectaline (3). The isolates were submitted to in vitro evaluation of lipoperoxidation (LPO) and cyclooxygenase enzymes (COX-1 and -2) inhibitory properties and showed moderate antioxidant activities (40-70%) at 100 ppm when compared to commercial standards BHT and vitamin E and moderate inhibition of COX-1 (ca. 40%) and marginal inhibition of COX-2 enzymes (< 10%) at 100 ppm when compared to nonsteroidal anti-inflammatory drugs (NSAIDs) aspirin, rofecoxib, and celecoxib, respectively.

Atrazine interaction with tropical humic substances by Enzyme Linked Immunosorbent Assay

Enzyme-Linked Immunosorbent Assay (ELISA) has been evaluated by analyzing rich-humic water samples from tropical rivers. The samples were spiked with atrazine at ppb level Different pHs (4 to 9) and humic concentrations (2.5 to 40 mg L-1) were investigated. The assay performance showed a strong dependence on the pH values and amount of humic matter at low atrazine concentration. From all the conditions studied the low pH (pH 4) and high humic substances concentrations (40 mg L-1) showed the greatest influence. The IC50 value to control sample (no humic) diminished from 0.28 nmol L-1 to 0.64 nmol L-1 to humic acid solution. This effect is specially noted for the humic acid fractions, since fulvic acid fractions showed no significant change on the immunoassay results. Additionally, it has been demonstrated that at basic pH the matrix effect produced by the natural Brazilian water sample containing humic substances even at 40 mg L-1 disappears. Therefore, the ELISA method used to determine atrazine, can be employed to determine this pesticide in water samples containing humic substances without prior preparation.

CENTRAL ANGIOTENSIN-CONVERTING ENZYME-BLOCKADE AND THIRST

1. The effect of lisinopril, a potent inhibitor of angiotensin converting enzyme (ACE), injected into the medial preoptic area (MPOA) on water intake was investigated in male Holtzman rats (200-250 g).2. Injection of lisinopril (2 mug/mul) into the MPOA abolished the water intake induced by subcutaneous (sc) injection of isoprenaline (100%) and water deprivation (90%) and drastically reduced the water intake induced by sc injection of polyethyleneglycol (60%). A small reduction of water intake induced by lisinopril was also observed 90 and 120 min after sc hypertonic saline (N = 10 for each group).3. These results suggest that central ACE activation, particularly in the MPOA, plays an important role in the dipsogenic responses induced by the agents studied.

Activity of Delta(9)-desaturase enzyme in mammary gland of lactating buffaloes

The objective of this research was to measure the activity of e-desaturase enzyme in lactating buffaloes. Data from forty lactating Murrah-crossbred buffaloes were collected on five commercial farms located at Sarapui and Pilar do Sul, São Paulo-Brazil. A field survey was done from April to November 2002. In four farms, buffaloes were fed with wet brewers grains (primary concentrate). Only one farm (Farm 4) offered pasture and corn silage. Monthly milk samples were collected and stored at -20 degrees C until analyzed for fatty acid composition. The Delta(9)-desaturase activity was measured using an indirect method (myristoleic and myristic acids ration - C(14:1c9)/C(14:0)). The higher C(14:1c9)/C(14:0) rate was verified on Farm 4 (0.092). The C(14:1c9)/C(14:0) ratio were 0.064 to Farm 1; 0.065 to Farm 2; 0.062 to Farm 3 and 0.065 to Farm 5. The C(17:1)/C(17:0), C(18:1c9)/C(18:0) and C(18:2c9t11)/C(18:1t11) ratios were also affected. The Farm 4 showed higher value for all ratios. Therefore, in lactating buffaloes grazing pasture the Delta(9)-desaturase activity could be enhanced.

Morphofunctional changes in Leydig cells throughout the continuous spermatogenesis of the freshwater teleost fish, Serrasalmus spilopleura (Characiformes, Characidae): an ultrastructural and enzyme study

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

An enzyme-linked immunosorbent assay (ELISA) for detection of antibodies against Toxocara vitulorum in water buffaloes

Toxocara vitulorum, a parasite of the small intestine of cattle and water buffaloes, is mainly acquired by calves via the colostrum/milk from infected cows. To understand the development of immune responses in calves, antibody levels to a soluble extract antigen (Ex) from T. vitulorum infective larvae were measured by an indirect ELISA with sera of 15 buffalo calves, which were sampled every 15 days for the first 180 days after birth and 9 buffalo cows during the perinatal period. From all serum samples examined during the first 180 days, antibody level was lowest and highest in calves at 1 day of age before and after suckling colostrum, respectively, suggesting that the origin of antibodies was the colostrum. Immediately after birth, antibody levels in suckled calves remained at high levels until day 15, began to decrease to lower levels between 15 and 30 days and remained relatively stable until 120 days. By comparing the immune responses of these animals with their parasitological status it was considered possible to determine if passively acquired or actively produced antibodies provided protection against the infection. High numbers of T. vitulorum eggs in the feces between 30 and 60 days indicated that passively acquired antibodies did not provide protection against the infection, at least during these first days, and the maximum fecal egg counts during 30-45 days were coincident with decreased antibody levels. Between 60 and 120 days, when serum antibodies were detected at reduced, but stable levels, adult nematodes were expelled from the intestines and no more T. vitulorum eggs were found, suggesting development of acquired resistance. However, the potential and functional protective role of the antibodies against T. vitulorum infection and the process of self-cure requires further investigation. (C) 2001 Elsevier B.V. B.V. All rights reserved.

Effect of enzyme supplementation of broiler diets based on corn and soybeans

Digestibility of diets based on corn and soybean meal or soybeans treated by roasting or extrusion, with or without an enzyme supplementation, was measured by true (Sibbald) methods, by analysis of excreta, and by analysis of ileal digesta. Only analysis of ileal digesta was able to consistently measure differences between soybean and enzyme treatments in the digestibility of CP, starch, fat, and ME. The amino acid (AA) digestibility of the diets was measured by analysis of the ileal contents. Whereas enzyme supplementation improved overall CP digestibility by 2.9%, this improvement was not equal for all AA. of the AA most important for broilers fed corn-soybean diets, the digestibilities of Lys, Met, and Arg were not improved or not improved significantly by the enzyme supplementation; however, that of Val was improved by 2.3% and that of Thr was improved by 3.0%. A performance trial demonstrated that enzyme supplementation with equal diet formulation improved BW and the feed conversion ratio by 1.9 and 2.2%, respectively. A second performance trial compared standard diet formulations with formulations using enzyme supplementation and energy levels that were reduced by the amount of improvement provided by the inclusion of enzyme in the first performance trial. No difference was seen between treatments, showing that the improvement of nutrient utilization brought about by enzyme supplementation completely compensated for the reduced energy content. Whereas enzyme supplementation should allow a reduction in CP formulation as well, individual AA were not improved equally by supplementation and should also be balanced.