RESUMO

Didelphis albiventris Lund, 1840 is one of the least known Brazilian marsupials with respect to its feeding habits. Since the white-eared opossum is omnivorous, it occupies several niches and may be found in urban areas as well as in forest fragments. Current analysis aimed at determining and comparing the diet of D. albiventris in two periurban areas of the municipality of Viamão, a conserved forest remnant and a degraded area resulting from agricultural activities. Captures of D. albiventris were carried out for a period of 30 months by a sampling effort of 504 trap-nights. Forty fecal samples were collected from 45 captures corresponding to 18 specimens.  Diet was determined by fecal analysis. Fruit was the most representative item, at an occurrence of 77.5%, comprising six species of plants, followed by invertebrates (62.5%) and vertebrates (37.5%). Didelphis albiventris displayed a frugivorous-omnivorous habit, with no difference between the sexes with respect to composition and frequency of the items consumed. Since there was a greater diversity of food items and fruits in the more conserved area, seasonal variations in the diet could be detected. Generalist species, such as D. albiventris, may have an important role in the recovery of degraded areas through the dispersal of seeds of species of initial successional stages.(AU)

Didelphis albiventris Lund, 1840 é um dos marsupiais brasileiros menos conhecidos quanto aos hábitos alimentares. De hábito onívoro, ocupa uma variedade de nichos, sendo encontrado tanto em áreas urbanas quanto em fragmentos florestais. Teve-se como objetivo conhecer e comparar a dieta de D. albiventris em duas áreas periurbanas do município de Viamão: um remanescente florestal conservado e uma área degradada por atividades agropastoris. Foram efetuadas capturas de D. albiventris durante um período de 30 meses, resultando em um esforço amostral de 504 armadilhas-noite. Foram coletadas 40 amostras de fezes decorrentes de 45 capturas correspondentes a 18 indivíduos. A dieta foi determinada através de análise fecal. Com ocorrência de 77,5%, os frutos foram o item mais representativo, compreendendo seis espécies de plantas, seguido de invertebrados, com 62,5%, e vertebrados, com 37,5%. Didelphis albiventris apresentou hábito frugívoro-onívoro, não havendo diferença entre os sexos quanto à composição e frequência dos itens consumidos. Na área mais conservada, obteve-se a mais alta diversidade de itens alimentares e de frutos, encontrando-se variações sazonais na dieta. Espécies generalistas, tais como D. albiventris, podem desempenhar importante papel na recuperação de áreas degradadas, efetuando a dispersão de sementes de espécies de estádios sucessionais iniciais.(AU)

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High amounts of albumin in urine cause tubulointerstitial damage that leads to a rapid deterioration of the renal function. Albumin exerts its injurious effects on renal cells through a process named endoplasmic reticulum (ER) stress due to the accumulation of unfolded proteins in the ER lumen. In addition, albumin promotes phosphorylation and consequent activation of MAPKs such as ERK1/2. Since ERK1/2 activation promoted by albumin is a transient event, the aims of the present work were to identify the phosphatase involved in their dephosphorylation in albumin-exposed cells and to analyze the putative regulation of this phosphatase by albumin. We also sought to determine the role played by the phospho/dephosphorylation of ERK1/2 in the cellular response to albumin-induced ER stress. MAP kinase phosphatase-1, MKP-1, is a nuclear enzyme involved in rapid MAPK dephosphorylation. Here we present evidence supporting the notion that this phosphatase is responsible for ERK1/2 dephosphorylation after albumin exposure in OK cells. Moreover, we demonstrate that exposure of OK cells to albumin transiently increases MKP-1 protein levels. The increase was evident after 15 min of exposure, peaked at 1 h (6-fold) and declined thereafter. In cells overexpressing flag-MKP-1, albumin caused the accumulation of this chimera, promoting MKP-1 stabilization by a posttranslational mechanism. Albumin also promoted a transient increase in MKP-1 mRNA levels (3-fold at 1 h) through the activation of gene transcription. In addition, we also show that albumin increased mRNA levels of GRP78, a key marker of ER stress, through an ERK-dependent pathway. In line with this finding, our studies demonstrate that flag-MKP-1 overexpression blunted albumin-induced GRP78 upregulation. Thus, our work demonstrates that albumin overload not only triggers MAPK activation but also tightly upregulates MKP-1 expression, which might modulate ER stress response to albumin overload.

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The resistance of the opossum Didelphis aurita to Bothrops snake venoms is attributed to the opossum's antihemorrhagic (DM43) and antimyotoxic (DM64) acidic serum glycoproteins. The aim of this study was to characterize the N-glycosylation sites of these antiophidic proteins and to determine whether their glycans influence the biological activity measured by in vitro assays. Our experimental pipeline included the sequential enzymatic digestion of the inhibitors with two different proteinases (trypsin and endoproteinase Asp-N) and eventually with trypsin, peptide-N-glycosidase F (PNGase F) and endoproteinase Asp-N, used in that order. All of the peptide and protein samples were analyzed by MALDI-TOF/TOF MS. The results experimentally confirmed the putative N-glycosylation sites of DM43 (Asn23, Asn156, Asn160, and Asn175) and DM64 (Asn46, Asn179, Asn183, and Asn379). Following treatments with specific glycosidases, complex-type oligosaccharides containing galactose and sialic acid could be assigned to both proteins. The removal of these monosaccharide units by exoglycosidase digestion did not measurably affect the inhibitory activity. In contrast, partially deglycosylated DM43 treated with PNGase F under nondenaturing conditions was half as effective as native DM43. In conclusion, we have demonstrated that the contribution of the carbohydrate portion of these potentially therapeutic molecules, for their mechanism of action, should not be overlooked.

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The aim of this study was to identify and quantify the argyrophil, argentaffin and insulin-immunoreactive cells (IIC) in the small intestine of the opossum Didelphis aurita. Seven adult male specimens of opossums were investigated. The animals were captured, and their blood insulin levels were determined. After euthanasia, fragments of the small intestine were processed for light microscopy and transmission electron microscopy, and submitted to histochemistry and immunohistochemistry for identification of argyrophil and argentaffin endocrine cells, and IIC. Argyrophil and argentaffin cells were identified in the intestinal villi and Liberkühn crypts, whereas IIC were present exclusively in the crypts. Ultrastructure of the IIC revealed cytoplasmic granules of different sizes and electron densities. The numbers of IIC per mm(2) in the duodenum and jejunum were higher than in the ileum (p<0.05). The animals had low levels of blood insulin (2.8 ± 0.78 µIU/ml). There was no correlation between insulin levels and the number of IIC in the small intestine. The IIC presented secretory granules, elongated and variable morphology. It is believed that insulin secretion by the IIC may influence the proliferation of cells in the Liberkühn crypts, and local glucose homeostasis, primarily in animals with low serum insulin levels, such as the opossum.

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