Investigation of the interaction between Tc85-11 protein and antibody anti-T cruzi by AFM and amperometric measurements

This present work reports on development of an amperometric immunosensor for the diagnosis of Chagas' disease using a specific glycoprotein of the trypomastigote surface, which belongs to the Tc85-11 protein family of Trypanosoma cruzi (T cruzi). An atomically flat gold surface on a silicon substrate and gold screen-printed electrodes were functionalized with cystatrine and later activated with glutaraldehyde (GA), which was used to form covalent bonds with the purified recombinant antigen (Tc85-11). The antigen reacts with the antibody from the serum, and the affinity reaction was monitored directly using atomic force microscopy or amperometry through a secondary antibody tagged to peroxidase (HRP). Surface imaging allowed to us to differentiate the modification steps and antigen-antibody interaction allowed to distinguish the affinity reactions. In the amperometric immunosensor, peroxidase catalyses the L-2 formation in the presence of hydrogen peroxide and potassium iodide, and the reduction current intensity was measured at a given potential with screen-printed electrodes. The immunosensor was applied to sera of chagasic patients and patients having different systemic diseases. (c) 2006 Elsevier Ltd. All rights reserved.

Immunosensor for the diagnosis of Chagas' disease

Trypanosoma cruzi proteins from epimastigote membranes, herein referred as antigens, have been used for the construction of an amperometric immunosensor for serological diagnosis of Chagas' disease. The proteins used had a molecular mass ranging from 30 to 100 kDa. The gold electrode was treated with cysteamine and glutaraldehyde prior to antigen immobilization. Antibodies present in the serum of patients with Chagas' disease were captured by the immobilized antigens and the affinity interaction was monitored by chronoamperometry at a potential of -400 mV (versus Ag pseudo-reference electrode) using peroxidase-labeled IgG conjugate and hydrogen peroxide, iodide substrate. The incubation time to allow maximum antigen-antibody and antibody-peroxidase-labeled IgG interactions was 20 min with a reactivity threshold at -0.104 mu A. (c) 2004 Elsevier B.V. All rights reserved.

Synthesis and in vitro evaluation of potential antichagasic hydroxymethyinitrofurazone (NFOH-121): A new nitrofurazone prodrug

The synthesis of mutual prodrugs of nitrofurazone with primaquine, using specific and nonspecific spacer groups, has been previously attempted seeking selective antichagasic agents. The intermediate reaction product, hydroxymethylnitrofurazone (NFOH-121), was isolated and tested in LLC-MK2 culture cells infected with trypomastigotes forms of Trypanosoma cruzi showing higher trypanocidal activity than nitrofurazone and benznidazol in all stages. The mutagenicity tests showed that the prodrug was less toxic than the parent drug. Degradation assays were carried out in pH 1.2 and 7.4. (C) 2003 Elsevier Ltd. All rights reserved.

EFEITOS DOS MÉTODOS DE ABATE DE BOVINOS NA EFICIÊNCIA da SANGRIA

O objetivo do presente trabalho foi avaliar os efeitos dos métodos de abate de bovinos com insensibilização prévia através de marreta e pistola pneumática de penetração e sem insensibilização, realizada pelo método kasher, na eficiência da sangria. A eficiência da sangria foi avaliada pela determinação da hemoglobina no sangue e na carne. Foram utilizados 240 bovinos sãos, da raça Nelore, abatidos em matadouro-frigorífico sob Serviço de Inspeção Federal, sendo 60 animais abatidos pelo método judaico (kasher), 60 animais insensibilizados através de marreta, 60 animais insensibilizados através de pistola pneumática de penetração e 60 animais insensibilizados através de pistola pneumática de penetração seguida por estimulação elétrica. Foram colhidas amostras de sangue após a sangria dos animais e de carne, na porção torácica do músculo Longus colli, imediatamente após a chegada da carcaça à câmara frigorífica. Os animais abatidos pelo método kasher apresentaram menor teor de hemoglobina residual na carne e melhor eficiência da sangria. A utilização da estimulação elétrica imediatamente após a sangria dos animais atordoados por pistola pneumática, afetou negativamente a eficiência da sangria.

Biodisponibilidade de ferro em diferentes compostos para leitões desmamados aos 21 dias de idade

Avaliou-se a biodisponibilidade de ferro de diferentes compostos visando sua utilização em dietas para leitões desmamados. Utilizaram-se 44 leitões (7 não-anêmicos e 37 anêmicos) desmamados aos 21 dias de idade (7,3 ± 1,8 kg) e distribuídos em dois grupos: grupo não-anêmico e grupo anêmico. Durante sete dias, os animais do grupo não-anêmico receberam dieta com FeSO4.7H2O (sulfato ferroso hepta-hidratado) na dose de 100 mg/kg e os do grupo anêmico, dieta sem ferro (<15 mg/kg ração). No sétimo dia, depois de determinada a concentração de hemoglobina sanguínea e diagnosticada a anemia, os leitões foram agrupados segundo o produto do peso (kg) × hemoglobina (g/dL) e alojados individualmente, durante 13 dias, em gaiolas para estudos de digestibilidade, onde foram alimentados com seis rações à base de milho e leite em pó: três rações-padrão com FeSO4.7H2O em quantidade equivalente a 80, 150 e 200 mg Fe/kg de ração; duas rações experimentais com ferro (150 mg/kg) na forma de FeSO4 microencapsulado com carboximetilcelulose ou de ferro quelado com metionina; e uma controle com ferro (100 mg/kg). O consumo de ração foi medido diariamente. Nos dias 0, 3, 6, 9 e 13 do período de repleção, os animais foram pesados para avaliação do desempenho e o sangue foi coletado para determinação da concentração de hemoglobina. Ao final do ensaio, os animais foram sacrificados e o fígado foi coletado para determinação das concentrações de ferro total, ferro heme e ferro não-heme. As concentrações hepáticas de ferro heme, ferro não-heme e ferro total não diferiram entre os animais, entretanto, os do grupo controle apresentaram excesso de ferro total no fígado, relacionado à dose de ferro injetada nos leitões após o desmame. em comparação ao FeSO4.7H2O não encapsulado, os compostos de ferro microencapsulado com carboximetilcelulose e de ferro quelado com metionina promovem melhor conversão alimentar em leitões desmamados.

Integrating geo-referenced multiscale and multidisciplinary data for the management of biodiversity in livestock genetic resources

P>In livestock genetic resource conservation, decision making about conservation priorities is based on the simultaneous analysis of several different criteria that may contribute to long-term sustainable breeding conditions, such as genetic and demographic characteristics, environmental conditions, and role of the breed in the local or regional economy. Here we address methods to integrate different data sets and highlight problems related to interdisciplinary comparisons. Data integration is based on the use of geographic coordinates and Geographic Information Systems (GIS). In addition to technical problems related to projection systems, GIS have to face the challenging issue of the non homogeneous scale of their data sets. We give examples of the successful use of GIS for data integration and examine the risk of obtaining biased results when integrating datasets that have been captured at different scales.

Oxytocin promotes bone formation during the alveolar healing process in old acyclic female rats

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Evaluation of different media for the production of cephalosporins by Streptomyces clavuligerus ATCC 27064

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

Influência do banho de aspersão ante-mortem na contaminação microbiana da carne bovina

To determine the effects of the pre-slaughter showering on some meat quality parameters, the bacterial changes in the Longus colli muscle and the contamination of the meat surface at three different points of the slaughter line were studied. Sixteen Nelore steers were slaughtered in a commercial slaughterhouse. Eight animals were submitted to pre-slaughter showering; a control group of eight animals were slaughtered without showering. Aseptic samples were collected for evaluations in the muscle depth, in the anterior portion of Longus colli muscle, just before chilling. The swab method was used for sampling carcass surface right after dressing, before carcasse washing, and at the beginning of chilling. Longus colli muscle samples were used to determine bacteria total count, psychrotrophic count and Enterobacteriaceae count, after 5, 24 and 48 hours from slaughtering, and in carcass surface, bacteria total count and psychrotrophic count. Multivariate methods were used to evaluate bacterial data the use of pre-slaughter showering did not affect the bacteria total counts, in the deep tissue. A significant growth of psychrotrophic bacteria was detected in both treatments. No significant differences (P>.05) were found in bacteria total count and psychrotrophic count between treatments. Also, no differences (P>.05) were detected between counts taken at different momments at the kill floor: skinning, before carcasse washing, and at the cooler, before chilling.

Synthesis and in vitro evaluation of potential antichagasic dipeptide prodrugs of primaquine

American trypanosomiasis (Chagas' disease) is an endemic parasitic disease afflicting more than 20 million people in Latin America. Currently, therapy is unsatisfactory and only two drugs are available. Primaquine, an antimalarial drug, has trypanocidal activity. Dipeptide derivatives of primaquine, Phe-Arg-PQ, Lys-Arg-PQ, and Phe-Ala-PQ, were synthesized. The choice of the peptides was based on the primary specificity of cruzipain, the major cysteine proteinase from T. cruzi. The prodrugs obtained were tested on the LLC-MK2 cell culture infected with trypomastigotes forms of T. cruzi Phe-Arg-PQ, Lys-Arg-PQ, and Phe-Ala-PQ were active in all stages.

Influence of free amino acids on clavulanic acid production by Streptomyces clavuligerus in synthetic medium

The effect of nitrogen source on clavulanic acid production was investigated in shake flasks. Media containing asparagine plus one of several different amino acids or a combination of two amino acids was tested. The best result, ca. 180 mg/L clavulanic acid, CA, in 60 h, was obtained with the lysine-tyrosine pair In an aerated and agitated fermentor this medium led to CA concentrations of ca. 210 mg/L, a remarkable production for synthetic medium utilization. Amino acids analysis during cultivation indicated that, while asparagine was consumed rapidly, lysine and tyrosine were metabolized slowly, promoting CA production.

Study of spore germination in Thelypteris-dentata (forssk) e-st-john

The spore germination in Thelypteris dentata was studied and the optimum temperature of 25 C was determined by the maximum percentage germination and the highest germination index. The necessity of continuous white light irradiation and reversion of spore germination by subsequent far-red light irradiation indicate the involvement of phytochrome in the control of the process. The dependence of induction of spore germination to the continuous white light low irradiance confirms the phytochrome control through the low fluence response.

Influência do banho de aspersão ante-mortem na eficiência da sangria e em parâmetros bioquímicos da carne bovina

To determine the effects of the pre-slaughter showering on some meat quality parameters, the biochemical changes in the Longus colli muscle and the bleeding efficiency were studied. Thirty-six Nelore steers were slaughtered in a commercial slaughterhouse. Eighteen animals were submitted to pre-slaughter showering; a control group of eighteen animals were slaughtered without showering. Samples were collected for evaluations in the muscle depth, in the anterior portion of longus colli muscle,just before chilling. Bleeding efficiency was evaluated through the ratio of muscle haemoglobin/blood haemoglobin using blood samples taken five seconds after bleeding, and muscle sample taken before chilling. Longus colli muscle samples were also used to determine glycogen, glucose, pH and acidity, 5, 24 and 48 hours after slaughtering. Multivariate methods were used to evaluate biochemical data and the bleeding efficiency data analysis followed the randomized block design. Haemoglobin retained in the muscle and bleeding efficiency were not affected (P > .05) by pre-slaughter showering. The pre-slaughter showering did not affect (P > .05) the glycolysis. There was a significant effect of time in glycogen, glucose, pH and acidity, in the first 24 hours.