Renal dysfunction in dogs envenomed by snakes has mostly been evaluated using routine serum and urine renal markers. Generally, these are not able to detect the renal damage at an early stage and their sensitivity is affected by hemolysis, hematuria and pigmenturia. Selective use of urinary biomarkers can provide early information on severity and stage of renal injury caused by nephrotoxic substances before major decline in renal function occurs. Therefore, the aim of this study was to evaluate venom-induced renal damage using urinary markers of glomerular (urinary albumin (uAlb), immunoglubin G (IgG) and Creactive protein (uCRP)) and proximal tubular dysfunction (urinary retinol binding protein (uRBP)) and compare these markers with routine renal markers (serum urea (BUN) and creatinine (sCr), urinary specific gravity (USG) and urinary protein to creatinine ratio (UPC)).Nineteen dogs envenomed by either neurotoxic or cytotoxic snakes and ten clinically healthy dogs were included in this study. Urinary markers were measured using previously validated commercially available ELISA kits. Among measured routine renal markers a significant difference between snake -envenomed and healthy dogs was noted only in UPC, but in the presence of hematuria and hemoglobinuria, differentiation between prerenal and renal proteinuria was not possible. The urinary biomarkers uAlb, uIgG and uRBP in snakeenvenomed dogs were significantly increased (P<0.05) when compared to healthy dogs at admission, whereas 24 h after envenomation only uCRP was significantly elevated. Using urinary /markers, results of this study showed that snake venom evokes renal dysfunction at the glomerular and tubular region of the nephron.

We investigated the influence of SORCS1 polymorphisms on insulin secretion in obese women with PCOS. Metabolic status was recorded in 50 clinically well characterized PCOS patients. Oral glucose tolerance test was performed and laboratory parameters of insulin resistance measured. All patients were genotyped for SORCS1 rs1358030, rs1416406 and rs11192966 polymorphisms. Statistical analysis was performed using the Mann-Whitney test. SORCS1 rs1416406 significantly influenced stimulated glucose plasma levels (p = 0.006) and increased glucose stimulated insulin secretion (p = 0.034). None of the polymorphisms influenced insulin resistance as measured by homeostatic model assessment. We report for the first time the relevance of SORCS1 polymorphisms for glycemic control and glucose stimulated insulin secretion in obese women with PCOS.

BackgroundThyroid hormone supplementation anecdotally has been described as a valid treatment option for dogs with aggression‐related problems. However, prospective, controlled, and blinded trials evaluating behavior and neurohormonal status in hypothyroid dogs during treatment with levothyroxine are lacking.ObjectiveLevothyroxine supplementation will have a significant influence on the behavior and neurohormonal status of dogs with spontaneous hypothyroidism.AnimalsTwenty client‐owned dogs diagnosed with spontaneous hypothyroidism.MethodsThis prospective study was to evaluate the behavior of dogs, which was screened at initial presentation, and after 6 weeks, and 6 months of treatment with levothyroxine (starting dosage 10 μg/kg PO q12h) using the standardized Canine Behavioral Assessment and Research Questionnaire (C‐BARQ). At each time period, circulating serotonin and prolactin (PRL) concentrations were evaluated using a commercially validated ELISA kit and heterologous radioimmunoassay, respectively.ResultsAfter 6 weeks of thyroid hormone supplementation, C‐BARQ scores demonstrated a significant increase in activity of hypothyroid dogs (P < .01). No significant change in any of the behavioral signs was observed after 6 months of treatment. No significant difference in circulating concentrations of serotonin (P > .99 and P = .46) and PRL (P = .99 and P = .37) were noted between the 6‐week and 6‐month periods compared with baseline.Conclusions and Clinical ImportanceThe results of this study indicate increased activity of hypothyroid dogs after 6 weeks of thyroid hormone supplementation. None of the hypothyroid dogs in this cohort showed a significant change in any of the evaluated behavioral signs and neurohormonal status after 6 months of thyroid hormone supplementation.

Abbreviations used: 7TM -seven transmembrane receptor; B max -max. receptor number; BSA -bovine serum albumin; Ct -threshold cycle; DMEM -Dulbecco's modified Eagles's Medium; DNA -deoxyribonucleic acid; D-PBS -Dulbecco's phosphate-buffered saline; ELISA -enzyme-linked immunosorbent assay; F-actin -filamentous actin; FCAflow cytometry analysis; FCS -flow cytometry sorting; GFP -green fluorescent protein; GPCR -G-protein coupled receptor; HA -hemagglutinin; HANK1-R -N-terminally HAtagged human neurokinin-1 receptor; HEK-293 -human embryonic kidney cells; HIFCSheat inactivated fetal calf serum; HRP -horseradish peroxidase; IC 50 -50% inhibitory concentration; IP1 -inositol phosphate 1; mRNA -messenger ribonucleic acid; NK1-Rneurokinin type 1 receptor; pEYFP -enhanced yellow fluorescent protein; PN -passage number; RT -reverse transcription; RT-PCR -quantitative real-time reverse transcription / polymerase chain reaction; SP -substance P; TMB -tetramethylbenzidine; WT -wildtype Abstract: This study focused on establishing and making a comprehensive functional characterization of an HEK-293-transfected cell line that would coexpress the enhanced yellow fluorescent protein-actin (pEYFP-actin) construct and the neurokinin type 1 receptor (NK1-R), which is a member of the seven transmembrane (7TM) receptor family. In the initial selection procedure, the cloning ring technique was used alone, but failed to yield clones with homogenous pEYFP-actin expression. Flow cytometry sorting (FCS) was subsequently used to enrich the pEYFP-actin-expressing subpopulation of cells. The enzyme-linked immunosorbent assay (ELISA), FCS and quantitative realtime reverse transcription/polymerase chain reaction (RT-PCR) were then employed to monitor the passage-dependent effects on transgene expression and to estimate the total β-actin/pEYFP-actin ratio. NK1-R was characterized via radioactive ligand binding and the second messenger assay. The suitability of the pEYFP-actin as a marker of endogenous actin was assessed by colocalizing pEYFP-actin with rhodamine-phalloidine-stained F-actin and by comparing receptor-and jasplakinolide-induced changes in the actin cytoskeleton organization. These experiments demonstrated that: i) both constructs expressed in the generated transfected cell line are functional; ii) the estimated pEYFPactin:endogenous β-actin ratio is within the limits required for the functional integrity of the actin filaments; and iii) pEYFP-actin and rhodamine-phalloidinestained F-actin structures colocalize and display comparable reorganization patterns in pharmacologically challenged cells.

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