E‐ADA activity in lymphocytes of an experimental model of pythiosis treated with immunotherapy

Pythiosis is a life‐threatening disease caused by the oomycete Pythium insidiosum. Some authors have suggested the involvement of a Th2‐like immune response in the infected host, which leads to extensive tissue damage. The switch from a Th2 to a Th1 response pattern is one hypothesis to explain the curative properties of immunotherapy. Taking into account the importance of immunotherapy for pythiosis treatment and the contribution of adenine nucleotides in the immunoregulation of the host, we evaluated the ecto‐adenosine deaminase (E‐ADA; EC 3·5.4·4) activity in lymphocytes from rabbits inoculated with P. insidiosum. Rabbits were inoculated with 1 milliliter of zoospores subcutaneously injected into the lateral thorax; after developing lesions, the rabbits received eight doses of immunotherapy. E‐ADA activity was measured in lymphocytes and the adenine nucleotides and adenosine levels were quantitatively determined in serum. Rabbits with characteristic lesions of pythiosis showed a decreased E‐ADA activity (82·36%), a decreased adenosine triphosphate concentration (54·04%) and a higher adenosine concentration (2·51 fold), when compared with controls, after 28 days of inoculation. However, after the immunotherapy, the rabbits showed an increase in the E‐ADA activity when compared with control (78·62%), contributing for the change in the immune response. Our results reinforce the hypothesis that the change from a Th2 to a Th1 immune response with the participation of the purinergic system could be responsible for the curative properties of immunotherapy. Copyright © 2012 John Wiley & Sons, Ltd.     

Morphological disparity and evolutionary rates of cranial and postcranial characters in sloths (Mammalia,Pilosa, Folivora)

Sloth morphological evolution has been widely studied qualitatively, with comparative anatomy and morpho-functional approaches, or through quantitative assessments of morphological variation using morphometrics. Only recently, however, have folivoran morphological disparity and evolutionary rates begun to be evaluated using discrete character data. Nonetheless, patterns of morphological evolution in separate character partitions have not been investigated, neither the relative influence of, on the one hand, phylogeny, and on the other, dietary and locomotory adaptations of sloths. Here we evaluate those patterns using a phylomorphospace approach, quantifying morphological disparity and evolutionary rates, and investigating possible drivers of morphological evolution for cranial and postcranial characters in Folivora. The evolution of the morphology in those partitions is associated with distinct patterns of disparity among clades and ecological groups, even though the two partitions do not differ substantially in overall evolutionary tempo. Historical processes shaped the morphological evolution of sloths more consistently than ecological ones, although changes in postcranial characters also seem to be associated with locomotory adaptations, in which morphological convergences were much more common. We also discuss important methodological trade-offs in investigations of partitioned datasets mostly composed of fossil taxa.     

Oestrus synchronization in sheep with progesterone-impregnated (MAP) intravaginal sponges and a prostagland in analogue

A double injection of PGF2α at two different dosage levels, 5 mg and 10 mg, eleven days apart, were compared with a combination of MAP-sponges and 5 mg or 10 mg PGF2α.The combination of MAP-sponges plus 10 mg PGF2α gave the best syn=chronization results (93,4%), as well as the best conception rate (84,9%) when ewes were inseminated on a fixed time basis 68 and 80 hours after treatment.     

NTPDase and 5′ ecto-nucleotidase expression profiles and the pattern of extracellular ATP metabolism in the Walker 256 tumor

In this study, we evaluated the NTPDases and ecto-5′-nucleotidase (CD73) expression profiles and the pattern of adenine nucleotide hydrolysis in rats submitted to the Walker 256 tumor model, 6, 10 and 15 days after the subcutaneous inoculation. Using RT-PCR analysis, we identified mRNA for all of the members of the ecto-nucleoside triphosphate diphosphohydrolase family investigated and a 5′-nucleotidase. By quantitative real-time PCR, Entpd1 (Cd39) and Entpd2 (Cd39L1) and CD73 were identified as the dominant genes expressed by the Walker 256 tumor, at all times studied. Extracellular adenine nucleotide hydrolysis by the Walker 256 tumor was estimated by HPLC analysis. Rapid hydrolysis of extracellular ATP by the tumor cells was observed, leading to the formation of adenosine and inosine in cells obtained from solid tumors at 6 and 10 days after inoculation. Cells obtained from solid tumors at 15 days of growth presented high levels of AMP and presented adenosine as a final product after 90 min of incubation. Results demonstrate that the presence of NTPDases and 5′-nucleotidase enzymes in Walker 256 tumor cells may be important for regulation of the extracellular adenine nucleotides/adenine nucleoside ratio, therefore leading to tumor growth.     

Procoagulant activity of mouse transformed cells: Different expression in freshly isolated or cultured cells

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