Use of aqueous extract of noni in extender for sheep semen freezing / Utilização do extrato aquoso de noni em diluente para congelação de sêmen ovino

The study aimed to evaluate the action of aqueous extract of noni in an extender for sheep semen freezing. Treatments differed in inclusion of aqueous extract of noni in the extender: T1 ˗ no addition; T2 ˗ 24µg/mL; T3 ˗ 72µg/mL; and T4 ˗ 120µg/mL. Ejaculates were collected, diluted in the four treatments, and frozen. After thawing, the semen was subjected to a thermoresistance test and evaluated for subjective motility, vigor, membrane integrity assessment by hypo-osmotic swelling test, live-dead assay, computer-assisted sperm analysis and the status of sperm capacitation and acrosome reaction. Data were subjected to ANOVA, and then to Student Newman Keuls's test at 5% significance level. In the thermoresistance test after two hours of incubation, motility in T4 (120µg/mL) was lower than in the other treatments, with no differences in the HoS test in either diluted semen or in the semen evaluated immediately post-thawing, while for the other times, treatments showed similar responses. Regarding the motility parameters, a difference was observed for progressive motility, curvilinear velocity, average path velocity, and amplitude of lateral head displacement. As to the sperm capacitation status, a difference was observed between treatments for the sperm capacitated with intact acrosome.(AU)

Este estudo teve como objetivo avaliar a ação do extrato aquoso de noni em diluente para congelação de sêmen de carneiro. Os tratamentos diferiram quanto à inclusão de extrato aquoso de noni ao meio diluidor em: T1˗ sem adição de extrato; T2˗ 24µg/mL ; T3- 72µg/mL e 120µg/mL. Por meio de vagina artificial, 16 ejaculados foram coletados, diluídos entre os quatro tratamentos e congelados. Após o descongelamento, o sêmen foi submetido ao teste de termorresistência e avaliado quanto à motilidade subjetiva, ao vigor espermático, à integridade de membrana pelo teste hiposmótico, bem como ao teste supravital, à análise de sêmen assistida por computador (CASA) e ao status de capacitação espermática e de reação acrossomal. Os dados foram submetidos a uma análise de variância, seguida pelo teste de Student-Newman-Keuls com 5% de significância. No teste de termorresistência, após duas horas de incubação, a motilidade do T4 (120µg/mL) apresentou-se inferior aos demais tratamentos. Não houve diferença significativa no teste HOS tanto para o sêmen diluído quanto para o sêmen avaliado imediatamente pós-descongelação; para as demais horas, os tratamentos apresentaram comportamento semelhante. Para os parâmetros de cinética, foi observada diferença estatística para motilidade progressiva, velocidade curvilinear, velocidade do percurso médio e amplitude de deslocamento lateral da cabeça. Quanto ao estado de capacitação espermática, observou-se diferença entre os tratamentos para espermatozoide capacitado com acrossomo intacto.(AU)

Desempenho de tilápias-do-nilo alimentadas com farelo da casca de pequi / Performance of Nile tilapia fed with bran made of pequi peel

Avaliou-se o desempenho de tilápias-do-nilo alimentadas com farelo da casca de pequi (Caryocar brasiliense). Foram utilizados 200 alevinos, com idade de 37 dias e peso corporal médio de 0,63±0,25g, distribuídos em delineamento inteiramente ao acaso, com quatro tratamentos ­ zero, 20, 40 e 60% de substituição de ração comercial por farelo da casca de pequi ­ e cinco repetições representadas por caixas de cloreto de polivinila com capacidade para 130L, contendo 10 peixes cada, totalizando 20 unidades experimentais. As características de desempenho avaliadas foram consumo de ração, peso corporal, ganho de peso, conversão alimentar, comprimento total e viabilidade criatória. A conversão alimentar ­ 1,96µ; 2,21µ; 2,63µ; 3,12µ - piorou linearmente com a inclusão do farelo de casca da pequi, enquanto as demais variáveis de desempenho não foram influenciadas pelos tratamentos. Conclui-se que a inclusão do farelo da casca de pequi na ração piora a conversão alimentar, sem alterar as demais variáveis de desempenho.

The performance of Nile tilapia fed with bran made of pequi peel was evaluated. Two hundred fingerlings, at 37 days of age and with mean body weight of 0.63±0.25 g, were distributed in a completely randomized design with four treatments ­0, 20, 40 and 60% of replacement of commercial diet with bran made of pequi peel ­with five repetitions represented by boxes of polyvinyl chloride (PVC) with capacity for 130 L, with 10 fish each, totalizing 20 experimental units. The performance characteristics evaluated were feed intake, body weight, weight gain, feed conversion, total length, and live viability. Feed conversion ­1.96µ; 2.21µ; 2.63µ; 3.12µ­ increased linearly with the inclusion of bran made of pequi peel, while the other performance variables were not influenced by treatments. The conclusion is that the inclusion of bran peel in the pequi diet worsened feed conversion, without changing other performance variables.

Effect of oral ingestion of an extract of the herb Uncaria tomentosa on the biodistribution of sodium pertechnetate in rats

The aim of the present study was to determine the effect of the oral ingestion of an extract of the herb Uncaria tomentosa (cat's claw) on the biodistribution of the radiobiocomplex sodium pertechnetate (Na99mTcO4) in rats. The animals (male Wistar rats, 2 months old, 180-220 g), were treated (1 mL) with an U. tomentosa extract (32 mg/mL, N = 5) or 0.9 percent NaCl solution (control, N = 5) for 7 days. After this period, Na99mTcO4 (3.7 MBq, 0.3 mL) was injected through the ocular plexus and after 10 min the rats were killed, the organs isolated and counted in a well-gamma counter. A significant (P < 0.05) alteration in Na99mTcO4 uptake i) from 0.57 ± 0.008 to 0.39 ± 0.06 percentATI/organ (P < 0.05) and from 0.57 ± 0.17 to 0.39 ± 0.14 percentATI/g (P < 0.05) was observed in the heart, ii) from 0.07 ± 0.02 to 0.19 ± 0.07 percentATI/g in the pancreas, and iii) from 0.07 ± 0.01 to 0.18 ± 0.07 percentATI/g (P < 0.05) in muscle after treatment with this extract. Although these results were obtained with animals, caution is advisable in the interpretation of the nuclear medicine examination when the patient is using this herb. This finding is probably an example of drug interaction with a radiopharmaceutical, a fact that could lead to misdiagnosis of the examination in clinical practice with unexpected consequences for the patient.

A high-copy T7 Escherichia coli expression vector for the production of recombinant proteins with a minimal N-terminal His-tagged fusion peptide

We report here the construction of a vector derived from pET3-His and pRSET plasmids for the expression and purification of recombinant proteins in Escherichia coli based on T7 phage RNA polymerase. The resulting pAE plasmid combined the advantages of both vectors: small size (pRSET), expression of a short 6XHis tag at N-terminus (pET3-His) and a high copy number of plasmid (pRSET). The small size of the vector (2.8 kb) and the high copy number/cell (200-250 copies) facilitate the subcloning and sequencing procedures when compared to the pET system (pET3-His, 4.6 kb and 40-50 copies) and also result in high level expression of recombinant proteins (20 mg purified protein/liter of culture). In addition, the vector pAE enables the expression of a fusion protein with a minimal amino-terminal hexa-histidine affinity tag (a tag of 9 amino acids using XhoI restriction enzyme for the 5'cloning site) as in the case of pET3-His plasmid and in contrast to proteins expressed by pRSET plasmids (a tag of 36 amino acids using BamHI restriction enzyme for the 5'cloning site). Thus, although proteins expressed by pRSET plasmids also have a hexa-histidine tag, the fusion peptide is much longer and may represent a problem for some recombinant proteins.

Genome features of Leptospira interrogans serovar Copenhageni

We report novel features of the genome sequence of Leptospira interrogans serovar Copenhageni, a highly invasive spirochete. Leptospira species colonize a significant proportion of rodent populations worldwide and produce life-threatening infections in mammals. Genomic sequence analysis reveals the presence of a competent transport system with 13 families of genes encoding for major transporters including a three-member component efflux system compatible with the long-term survival of this organism. The leptospiral genome contains a broad array of genes encoding regulatory system, signal transduction and methyl-accepting chemotaxis proteins, reflecting the organism's ability to respond to diverse environmental stimuli. The identification of a complete set of genes encoding the enzymes for the cobalamin biosynthetic pathway and the novel coding genes related to lipopolysaccharide biosynthesis should bring new light to the study of Leptospira physiology. Genes related to toxins, lipoproteins and several surface-exposed proteins may facilitate a better understanding of the Leptospira pathogenesis and may serve as potential candidates for vaccine.

Controle de qualidade externo do eritrograma: uma experiência em laboratórios de análises clínicas do Estado do Paraná / Quality control in hematology: an experience in clinical laboratories of Paraná State

Em experiência piloto de controle de qualidade externo para os valores do eritrograma, foi distribuída amostra constituída de eritrócitos humanos suspensos em meio de conservação (LEONART et all, Rev. Bras. Anál. Clín., 21:111-3, 1989) a 10 laboratórios de análises clínicas da Secretaria de Saúde do Estado do Paraná e a 4 outros laboratórios conceituados, incluindo os do Curso de Farmácia e do Hospital de Clínicas da Universidade Federal do Paraná. Os profissionais dos laboratórios participantes empregaram diferentes metodologias para a determinação de eritrócitos, hemoglobina, volume globular, volume corpuscular médio, hemoglobina corpuscular média e concentração de hemoglobina corpuscular média, seguindo instruções preconizadas para o manuseio da amostra. Os laboratórios demonstraram bom desempenho e os dados obtidos tiveram boa reprodutibilidade intra e interlaboratoriais. Estes resultados sugerem a viabilidade da amostra e do sistema empregado para implantação em Programas de Controle de Qualidade Interlaboratoriais.