The aim of this study was to evaluate the effect of negative energy balance (NEB) on the final growth of the dominant ovarian follicle in Holstein cows. Cows at 14 to 21 d postpartum from an intensive dairy farm were randomly selected and allocated into a positive energy balance group (PEB, with β-hydroxybutyric acid (BHBA) level < 1.2 mmol/L, n = 15) and an NEB group (BHBA > 1.2 mmol/L, n = 15). Plasma samples were collected at 21, 50 and 55 d postpartum to assess the concentrations of energy metabolites, minerals, hormones and cytokines. Ovaries were examined by transrectal ultrasound on days 50 and 55 (120 hours later) to evaluate the diameter of the largest follicle. Compared with PEB cows, there were a more severe body condition loss and a lower milk yield in NEB cows (P < 0.05) and these had greater concentrations of plasma BHBA, non-esterified fatty acids, triglycerides, urea nitrogen, growth hormone, interleukin 6, and fibroblast growth factor 21 and lesser concentrations of plasma glucose, total cholesterol, insulin, insulin-like growth factor 1, insulin-like growth factor binding protein 3, leptin, brain-derived neurotrophic factor and angiopoietin-like protein 8 on d 21 (P < 0.05), while plasma minerals were not affected by energy status (P > 0.05). These changes persisted until the end of the study period (50–55 days postpartum) resulting in a lower follicular growth rate for cows in the NEB than the PEB group. These observations indicate that follicular growth rate is associated with measurable changes in energy metabolite, hormone and cytokine concentrations caused by early postpartum NEB.

OBJECTIVE To develop a technique for isolation and culture of canine insulinoma cells and assess expression of cellular hexokinases (glucokinase and hexokinase I) and expression and secretion of insulin from these cells in vitro. SAMPLE Pancreatic insulinomas and normal pancreatic tissue from 4 and 3 dogs, respectively. PROCEDURES Tissues were collected by surgical excision or at necropsy. Insulinoma cells from 2 dogs were cultured for up to 10 weeks with standard techniques; insulin synthesis in vitro was confirmed by immunohistochemical analysis of freshly prepared slides of cultured cells, and insulin secretion was assessed by measurement of insulin concentrations in culture medium with an ultrasensitive mouse insulin ELISA. Expression of cellular hexokinases in insulinomas and adjacent normal (nontumor) pancreatic tissue from the same dog (n = 3) was examined by quantitative reverse transcriptase PCR assay. RESULTS Insulinoma cells survived for up to 10 weeks but did not proliferate in culture. Insulin was detected in isolated cells and secreted into culture medium for up to 10 weeks. Both cellular hexokinases were expressed; glucokinase appeared to be overexpressed in insulinomas, compared with normal pancreatic tissue from the same dogs. CONCLUSIONS AND CLINICAL RELEVANCE Canine insulinomas expressed hexokinases responsible for glucose responsiveness. Insulinoma cells were successfully maintained in short-term culture; cultured cells remained functional for 10 weeks as evidenced by cellular insulin content and had detectable secretion of insulin into the culture medium for ≥ 5 weeks. Apparent glucokinase overexpression by insulinomas suggested a possible mechanism underlying excessive insulin release by these tumors.

From 50 to 60% of companion animals in the United States are overweight or obese and this obesity rate is rising. As obesity is associated with a number of health problems, an agent that can help weight loss in pets and assist in clinically managing obesity through veterinary prescription foods and medication would be beneficial. Many studies have shown that celastrol, a phytochemical compound found in Celastrus orbiculatus extract (COE), has anti-obesity and anti-inflammatory effects, although these effects have not yet been determined in canine or canine-derived cells. The objective of this study was to investigate the effects of celastrol on the adipogenic differentiation and lipolysis of canine adipocytes. Primary preadipocytes were isolated from the gluteal region of a beagle dog and the primary adipocytes were differentiated into mature adipocytes by adipocyte differentiation media containing isobutylmethylxanthine, dexamethasone, and insulin. In a water-soluble tetrazolium (WST) assay, the cell viability of mature adipocytes was decreased after treatment with COE (0, 0.93, 2.32, and 4.64 nM celastrol) in a concentration-dependent manner, although preadipocytes were not affected. Oil Red O (ORO) staining revealed that COE inhibited the differentiation into mature adipocytes and lipid accumulation in adipocytes. In addition, treatment with COE significantly reduced triglyceride content and increased lipolytic activities by 1.5-fold in canine adipocytes. Overall, it was concluded that COE may enhance anti-obesity activity in canine adipocytes by inhibiting lipid accumulation and increasing lipolytic activity.