Molecular analysis of the RHD pseudogene by duplex real-time polymerase chain reaction.

OBJECTIVES: In this study, we aimed to present a strategy for the detection of the RHD pseudogene (RHDψ) based on a real-time polymerase chain reaction (PCR) assay. BACKGROUND: The D-negative phenotype is associated with many genetic alterations. In populations with African ancestry, this phenotype commonly results from the silent variant RHDψ. The evaluation of RHDψ is essential for correct inference of the RhD phenotype in order to avoid false-positive results. METHODS: We utilised a new method for the simultaneous detection of RHDψ and a fragment from exon 5 of the wild-type RHD gene based on duplex real-time PCR assay. RESULTS: The PCR assay allowed specific detection of RHDψ. There was complete agreement between the results generated by the new test and the results generated by molecular analysis performed using end-point PCR methods previously described. CONCLUSIONS: The assay developed is easy to execute and presents the potential for routine use at blood banks and other associated facilities where it is desired to determine the presence of RHDψ.

Desempenho de alevinos de tilápia do nilo alimentados com folha de mandioca desidratada na dieta / Performance of nile tilapia fingerlings fed dehydrated cassava leaf in the diet

O objetivo foi avaliar o crescimento e a viabilidade econômica de alevinos de tilápia do Nilo (Oreochromis niloticus), alimentados com diferentes níveis de farinha da folha de mandioca desidratada na ração (FFMD). Foram utilizados 80 alevinos distribuídos em 20 recipientes de 18L, constando de sistema de recirculação de água, durante um período de 51 dias. Cem exemplares, alimentados três vezes ao dia até a saciedade, foram distribuídos aleatoriamente em um delineamento experimental inteiramente casualizado, constituindo-se de 4 tratamentos (0, 5, 10 e 15% de adição da FFMD na ração) e 5 repetições cada. Os peixes alimentados com 10% de FFMD apresentaram melhor homogeneidade no crescimento. Entretanto, os indivíduos alimentados com inclusão de 5% de FFMD apresentaram homogeneidade de crescimento similar aos alimentados com 10%. Além disso, os peixes alimentados com 5% de FFMD obtiveram melhor desempenho zootécnico e viabilidade econômica da ração. Dessa maneira, recomenda-se a utilização de 5% da farinha de folha de mandioca desidratada em rações para tilápia do Nilo.

The aim was to evaluate the growth and the economic viability of the Nile tilapia (Oreochromis niloticus) fed with different levels of dehydrated cassava leaf flour in the diet (FFMD). 80 fingerlings were distributed into 20 18L containers, consisting of a water recirculation system over a period of 51 days. 100 copies fed three times a day to satiation were randomly assigned in a completely randomized design, constituting 4 treatments (0%, 5%, 10% and 15% addition of FFMD in diet) with 5 repetitions each. The fishes fed with 10% FFMD had better homogeneity in growth. However, individuals fed with inclusion of 5% FFMD homogeneity grew similarly to those fed with 10% FFMD. In addition, the fish from the 5% had better growth performance and economic viability of the feed. Thus, it is recommended to use 5% of dehydrated cassava leaf meal in diets for Nile tilapia.

Antioxidant and orofacial anti-nociceptive activities of the stem bark aqueous extract of Anadenanthera colubrina (Velloso) Brenan (Fabaceae).

The anti-nociceptive and antioxidant activities of the Anadenantheracolubrina stem bark aqueous extract (AEAC) were investigated. AEAC (30 µg/mL) reduced 94.8% of 2,2-diphenyl-1-picrylhydrazyl radical and prevented 64% (200 µg/mL) of lipid peroxidation caused by 2,2'-azobis(2-methylpropionamidine) dihydrochloride-induced peroxyl radicals. AEAC treatment (200 and 400 mg/kg) significantly (p < 0.001) reduced mice orofacial nociception in the first (61.4% and 62.6%, respectively) and second (48.9% and 61.9%, respectively) phases of the formalin test. Nociception caused by glutamate was significantly (p < 0.001) reduced by up to 79% at 400 mg/kg, while 56-60% of the nociceptive behaviour induced by capsaicin was significantly inhibited by AEAC (100-400 mg/kg). Mice treated with AEAC did not show changes in motor performance in the Rota-rod apparatus. It appears that AEAC is of pharmacological importance in treating pain due to its anti-nociceptive effects, which were shown to be mediated by central and peripheral mechanisms.