Pyometra-associated insulin resistance assessment by insulin binding assay and tyrosine kinase activity evaluation in canine muscle tissue.

Diestrus is associated with insulin resistance in bitches and pyometra can further impair insulin sensitivity. This study aimed to compare insulin sensitivity, insulin binding, and tyrosine kinase activity in bitches in anestrus, diestrus, or with pyometra. Patients submitted to elective ovariohysterectomy were divided into anestrus (n = 11) or diestrus (n = 13) according to reproductive history, vaginal cytology, and uterine histology. The group pyometra (n = 8) included bitches diagnosed with the disease based on clinical presentation and abdominal ultrasound findings and further confirmed by uterine histopathology. All patients were submitted to an intravenous glucose tolerance test (IVGTT) before ovariohysterectomy, and rectus abdominis muscle samples were collected during surgery for plasmatic membrane suspension preparation. Muscle-membranes were submitted to cold saturation insulin binding assay for dissociation constant (Kd) and maximum binding capacity (Bmax) determination, as well as exogenous substrate Poly (Glu: Tyr 4:1) phosphorylation assay for basal tyrosine kinase evaluation. Bitches with pyometra showed higher basal insulin (P < 0.001) and higher area under the curve (AUC) for insulin (P = 0.01) and glucose (P < 0.001) response during the IVGTT in comparison with bitches in anestrus or diestrus. Diestrus (P < 0.0001) and pyometra (P = 0.001) were associated with reduced tyrosine kinase activity in comparison with anestrus. No differences were documented in Kd and Bmax results for the low-affinity/high-capacity insulin receptors; however, high-affinity/low-capacity insulin receptors showed higher Kd and Bmax results in bitches in diestrus or with pyometra (P < 0.05) in comparison with anestrus. Despite the pyometra group showed the highest Kd values (P < 0.01), its Bmax results did not differ from the diestrus group (P > 0.05). Diestrus' higher Kd values and reduced tyrosine kinase activity in muscle tissue were compensated by increased total insulin binding capacity. Absent differences in IVGTT results between diestrus and anestrus bitches corroborate this finding. However, in bitches with pyometra, the highest Kd values were not compensated by increased total insulin binding capacity. This finding was associated with insulin resistance and glucose intolerance in IVGTT results. Moreover, pyometra resolution restored insulin sensitivity and glucose tolerance. These features can play a key role in pyometra-associated CDM, as well as in diabetic remission after pyometra resolution.

Protein and lipid metabolism adjustments in silver catfish (Rhamdia quelen) during different periods of fasting and refeeding / Ajustes no metabolismo de proteínas e lipídios no jundiá (Rhamdia quelen) durante diferentes períodos de jejum e realimentação

Abstract The fish may experience periods of food deprivation or starvation which produce metabolic changes. In this study, adult Rhamdia quelen males were subjected to fasting periods of 1, 7, 14, and 21 days and of refeeding 2, 4, 6, and 12 days. The results demonstrated that liver protein was depleted after 1 day of fasting, but recovered after 6 days of refeeding. After 14 days of fasting, mobilization in the lipids of the muscular tissue took place, and these reserves began to re-establish themselves after 4 days of refeeding. Plasmatic triglycerides increased after 1 day of fasting, and decreased following 2 days of refeeding. The glycerol in the plasma oscillated constantly during the different periods of fasting and refeeding. Changes in the metabolism of both protein and lipids during these periods can be considered as survival strategies used by R. quelen. The difference in the metabolic profile of the tissues, the influence of the period of fasting, and the type of reserves mobilized were all in evidence.

Resumo Os peixes podem sofrer períodos de privação de alimentos ou de fome, que produzem mudanças metabólicas. Neste estudo, jundiás machos adultos foram submetidos a jejum períodos de 1, 7, 14 e 21 dias e realimentação 2, 4, 6, e 12 dias. Os resultados demonstraram que a proteína do fígado foi esgotada depois de um dia de jejum, mas restabeleceu após 6 dias de realimentação. Após 14 dias de jejum, ocorreu a mobilização dos lípidos no tecido muscular sendo que estas reservas começaram a re-estabelecer-se após 4 dias de realimentação. Os triglicérides plasmáticos aumentam após um dia de jejum, e diminuiram após 2 dias de realimentação. O glicerol no plasma oscilou constantemente durante os diferentes períodos de jejum e realimentação. As alterações no metabolismo de proteína e lipídios durante estes períodos podem ser consideradas uma estratégias de sobrevivência utilizada pelo Rhamdia quelen. Sendo que a diferença no perfil metabólico tecidual bem como a influência do período de jejum e o tipo de reserva a ser mobilizada foram observadas neste estudo.

Redox and metabolic strategies developed by anterior and posterior gills of the crab Neohelice granulata after short periods of hypo- or hyper-osmotic stress.

The aim of this study was to identify the response pattern of redox balance, Na+/K+ATPase activity and HSP70 expression in the posterior and anterior gills of the crab Neohelice granulata submitted to hypo- or hyper-osmotic stress for 1 h and 6 h. After 1 h of either type of osmotic stress, there was an increase in catalase activity, but a decrease in GSSG/GSH ratio (oxidized to reduced glutathione ratio) and Na+/K+ATPase activity in both gill sets. H2O2 levels decreased only in the posterior gills. H2O2 levels and Na+/K+ATPase activity remained reduced after 6 h of exposure to either type of osmotic stress in both gill sets. The GSSG/GSH ratio returned to initial levels after 6 h of hyper-osmotic stress, whereas it increased 10 times in both gill sets after hypo-osmotic stress. Furthermore, HSP70 protein expression increased in posterior gills after 6 h of hypo-osmotic stress. H2O2 levels in tank water decreased after hypo-osmotic challenge and increased after 6 h of hyper-osmotic stress, indicating increased H2O2 excretion. Therefore, N. granulata gills have redox, metabolic and molecular strategies to deal with rapid osmotic challenges, an important environmental parameter that influences juvenile and adult crab distribution and abundance within different populations.