Development of a feline proinsulin immunoradiometric assay and a feline proinsulin enzyme-linked immunosorbent assay (ELISA): a novel application to examine beta cell function in cats.

The prevalence of feline diabetes mellitus has increased several-fold over the last three decades. In humans, progression from obesity to diabetes is marked by changes in the release of proinsulin. A specific proinsulin (FPI) assay has not been available to examine similar changes in cats. The goal of this study was to develop a proinsulin assay for the analysis of beta cell function in cats. Monoclonal antibodies were developed against recombinant FPI and used in a two-site sandwich immunoradiometric assay (IRMA) and enzyme-linked immunosorbent Assay (ELISA). The antibody pair had negligible cross-reactivity with bovine insulin and feline C-peptide. The working range was 11-667pmol/L for the IRMA and 11-1111pmol/L for the ELISA. An intravenous glucose tolerance test was performed in six long-term obese and six lean adult healthy cats and serum glucose, insulin, and FPI concentrations were determined. The proinsulin and insulin secretion pattern in response to glucose was significantly different between lean and obese cats but the pattern was similar within a group. Both groups had similar baseline proinsulin/insulin ratios; however, obese cats showed a significantly higher proinsulin/insulin ratio during the first 15min of the IVGTT and a much lower ratio during the last 30min suggesting a time-delayed adjustment to the increased insulin demand. In conclusion, we report the development and validation of an IRMA and an ELISA for FPI. This novel assay is useful to elucidate FPI secretion and can be used similar to a C-petide assay to evaluate residual beta cell function in cats.

Dyslipidemia in obese cats.

Obesity is an important endocrine disorder in cats and is a risk factor for diabetes similar to humans. The goal of this study was to examine the effect of long-term obesity and different diets (high protein, and high carbohydrate supplemented with saturated fatty acids or n-3 polyunsaturated fatty acids) on plasma lipids in the fasted and fed states in 12 lean (LEAN) and 12 obese (OBESE) cats with ultracentrifugation, and nuclear magnetic resonance spectroscopy. OBESE had higher plasma non-esterified fatty acids and triglycerides, as well as very-low-density-lipoproteins (VLDL) consisting primarily of medium-sized particles. The concentration of low-density-lipoproteins (LDL) was comparable between the groups, although OBESE had mostly very small, whereas LEAN had mostly large particles. The concentration of high-density-lipoproteins (HDL) was lower in OBESE and consisted primarily of small particles. Plasma triglycerides, and triglycerides and cholesterol in all lipoproteins increased postprandially. Different diets had little effect on lipids. Our results show that long-term obese cats develop similar lipoprotein changes to humans, yet, hypertension and atherosclerosis have not been described in obese cats. This suggests that dyslipidemia alone is not sufficient to induce hypertension and atherosclerosis. Other anti-atherogenic factors may be present in the obese, dyslipidemic cat.

Remission of diabetes mellitus in cats with diabetic ketoacidosis.

BACKGROUND: Diabetic ketoacidosis (DKA) has long been considered a key clinical feature of type-1 diabetes mellitus (DM) in humans although. An increasing number of cases of ketoacidosis have been reported in people with type-2 DM. HYPOTHESIS/OBJECTIVES: Cats initially diagnosed with DKA can achieve remission from diabetes. Cats with DKA and diabetic remission are more likely to have been administered glucocorticoids before diagnosis. ANIMALS: Twelve cats with DKA and 7 cats with uncomplicated DM. METHODS: Retrospective case review. Medical records of cats presenting with DKA or DM were evaluated. Diabetic remission was defined as being clinically unremarkable for at least 1 month after insulin withdrawal. The cats were assigned to 1 of 3 groups: (1) cats with DKA and diabetic remission; (2) cats with DKA without diabetic remission; and (3) cats with DM and diabetic remission. RESULTS: Seven cats with DKA had remission from diabetes. These cats had significantly higher concentrations of leukocytes and segmented neutrophils, and significantly lower concentrations of eosinophils in blood and had pancreatic disease more often than did cats with uncomplicated DM and diabetic remission. With regard to pretreatment, 3/7 cats in group 1, 1/5 cats in group 2, and 1/7 cats in group 3 had been treated with glucocorticoids. CONCLUSIONS AND CLINICAL IMPORTANCE: Remission of DM in cats presenting with DKA is possible. Cats with DKA and remission have more components of a stress leucogram, pancreatic disease, and seemed to be treated more often with glucocorticoids than cats with uncomplicated DM and diabetic remission.

Evaluation of IGF-1 levels in cats with transient and permanent diabetes mellitus.

It was investigated if IGF-1 levels in cats which experience diabetic remission (i.e. transient diabetes mellitus) differ from those in cats with permanent disease. Thirteen of 32 diabetic cats showed remission within 16 weeks after initiating insulin therapy, 19 cats continued to need insulin therapy. IGF-1 concentrations were measured before (t(0)), 1-3 (t(1)) and 4-8 (t(2)) weeks after initiating insulin therapy. No difference in IGF-1 levels was found between cats with transient and permanent diabetes at any point in time. In both groups of cats IGF-1 concentrations were significantly lower compared to those of controls before insulin administration. After starting insulin therapy IGF-1 increased significantly in both groups. In cats with transient diabetes IGF-1 levels were not different from controls already at t(1), whereas in cats with permanent diabetes it took until t(2). Although IGF-1 levels seem to normalize faster in cats with transient diabetes mellitus, measurement is not helpful to predict the course of the disease.

Evaluation of the low-dose dexamethasone suppression test and ultrasonographic measurements of the adrenal glands in cats with diabetes mellitus.

The objectives of the study were to evaluate the low-dose dexamethasone suppression (LDDS) test and the size of the adrenal glands via ultrasonography in cats with diabetes mellitus. Twenty-two cats were enrolled in the study. In 19 cats, suppression of cortisol concentrations below 5.5 nmol/litre occurred four and eight hours after intravenous administration of dexamethasone (0.1 mg/kg). In one other cat, the cortisol concentration was also below 5.5 nmol/litre at eight hours but was 11.0 nmol/litre at four hours. The results were in agreement with those of healthy cats in a previous study. The cortisol concentrations four and eight hours after administration of dexamethasone did not differ between cats with good glycemic control (n = 8) and those with moderate to poor control (n = 12). The adrenal glands of the diabetic cats were not enlarged compared with those of healthy cats. In two diabetic cats, the LDDS test results were abnormal. One cat had a pituitary adenoma and adrenal glands of normal size as determined by ultrasonography. The size of the adrenal glands of the other cat clearly differed; histological examination of the larger adrenal gland revealed an adrenocortical adenoma. Based on our findings, the results of the LDDS test using 0.1 mg/kg of dexamethasone are normal in cats with diabetes mellitus independent of the quality of glycemic control. In addition, diabetes mellitus does not lead to a measurable increase in the size of the adrenal glands in cats. Further studies are needed to evaluate if the dexamethasone dosage used in this study is useful to diagnose mild form of hypercortisolism.

Measurements of growth hormone and insulin-like growth factor 1 in cats with diabetes mellitus.

Serum concentrations of insulin-like growth factor 1 (IGF-1) and growth hormone were measured in 25 cats with untreated diabetes mellitus (11 of which were used for follow-up measurements, one to three, four to eight, nine to 12 and 13 to 16 weeks after their treatment with insulin began), 14 diabetic cats that had previously been treated with insulin, and seven diabetic cats that also had hypersomatotropism, two of which had not previously been treated with insulin; 18 healthy cats were used as controls. In the untreated diabetic cats the concentration of IGF-1 ranged from 13.0 to 433.0 ng/ml (median 170.5 ng/ml), which was significantly lower than the concentrations in the control cats (196.0 to 791.0 ng/ml, median 452.0 ng/ml). Their IGF-1 concentrations increased significantly when they were treated with insulin and after four to eight weeks were not different from those in the control cats. In the diabetic cats that had previously been treated with insulin the IGF-1 concentrations were 33.0 to 476.0 ng/ml (median 316.0 ng/ml), which was significantly lower than the concentrations in the control cats, but significantly higher than in the untreated diabetic cats. The IGF-1 concentrations in the two previously untreated diabetic cats with hypersomatotropism were low and low-normal but increased markedly after treatment with insulin. In the five previously treated cats with hypersomatotropism the concentration of IGF-1 was above the normal range. The concentrations of growth hormone in the treated and untreated diabetic cats without hypersomatotropisms were not significantly different and there was an overlap in its concentrations in the diabetic cats with and without hypersomatotropism.

[Diabetes mellitus in the cat: a review]. / Diabetes mellitus bei der Katze: Ein Uberblick.

About 80% of diabetic cats suffer from type 2 diabetes which is characterized by reduced insulin secretion from beta-cells and by insulin resistance. As in humans cats experienced a change in life habits and eating conditions over the last years leading to a tremendous increase in the prevalence of obesity. In both species obesity is one of the major risk factors for the development of type 2 diabetes. Treatment should be initiated immediately after diagnosis. In Zurich, therapy consists of application of an intermediate-acting insulin and dietary management. In the latter the use of diets with reduced carbohydrate content seems to be of utmost importance. We recently found that the percentage of cats with a transient course of diabetes increases from previous 25% to 50-70% when a diet with strongly reduced carbohydrate content is fed.

Evaluation of clinical signs and causes of lower urinary tract disease in European cats.

OBJECTIVES: To investigate the clinical signs and causes of lower urinary tract disease (LUTD) in 77 cats. METHODS: Cats diagnosed with LUTD over a two-year period were included in the study. RESULTS: The study population comprised 67 male and 10 female cats. Uroliths occurred in 17 of the 77 cats (22 per cent), urethral plugs in eight cats (10 per cent) and urinary tract infection in six cats (8 per cent). In 44 cats (57 per cent), no specific cause for the disease was found and they were classified as having idiopathic LUTD. In two of the 77 cats (3 per cent) no definitive diagnosis was established. Pain was less common in cats with uroliths and haematuria was more often seen in cats with urinary tract infection. At presentation, urethral obstruction was diagnosed in 45 of the 77 cats (58 per cent). CLINICAL SIGNIFICANCE: The causes of LUTD found in cats in this study are similar to those that have been previously documented, and idiopathic LUTD is the most frequent diagnosis. However, the rate of urethral obstruction, particularly in cats with idiopathic LUTD, was higher than in other reports. The cause of this difference is unknown.

Gene expression of glucose transporters and glycolytic enzymes in the CNS of rats behaviorally dependent on ethanol.

The steady-state levels of messenger RNA (mRNA) of the glucose transporters 1 and 3 and the glycolytic enzymes hexokinase, phosphofructokinase, glyceraldehyde-3-phosphate dehydrogenase and pyruvate dehydrogenase were measured in up to seven brain regions of the rat in a recently developed animal model of 'behavioral dependence' on ethanol. Irreversible behavioral dependence, including loss of control, was induced by offering the rats the choice between ethanol and water over a 9-month period (Group A). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet behaviorally dependent, Group B), a group forced to consume ethanol as sole fluid over a 9-month period (not behaviorally dependent, Group C) and ethanol-naive control rats. All groups were sacrificed 1 month after ethanol withdrawal. The mRNA concentrations of both neuronal glucose transporter 3 and the key glycolytic enzymes phosphofructokinase and pyruvate dehydrogenase were significantly reduced in the hippocampi of the rats behaviorally dependent on ethanol (Group A). No significant changes were seen in any of the remaining brain regions (e.g., cortical areas, limbic forebrain, amygdala, midbrain) in Group A, or in any brain area at all in Groups B and C. The results show that chronic consumption of ethanol in a free-choice situation may impair neuronal glucose uptake and glycolytic flux. This effect is manifested exclusively in the hippocampus and is specifically related to the development of behavioral dependence, since it was not found after forced administration of large amounts of ethanol (Group C).

Dopamine receptor gene expression in an animal model of 'behavioral dependence' on ethanol.

The steady-state levels of messenger RNA (mRNA) of five cloned dopamine (D) receptors were measured in five brain regions in rats in a recently developed animal model of 'behavioral dependence' on ethanol. One group of rats was given the choice between ethanol and water over a 9-month period and developed 'behavioral dependence' on ethanol (group a). This group was compared with a group given the choice between ethanol and water for only 2 months (not yet behaviorally dependent, group b), a group forced to consume ethanol as sole fluid over a 9-month period (not behaviorally dependent, group c) and ethanol-naive control rats. All groups were sacrificed 1 month after ethanol withdrawal. The concentrations of mRNA of D3-receptors in the limbic forebrain (which included the nucleus accumbens) were significantly lowered in groups a and b, but unchanged in group c. D3 mRNA levels were reduced in the hippocampus of group b and unchanged in the cortex, amygdala and striatum. No significant changes in the mRNA concentrations of D1-, D2-, D4- or D5-receptors were seen in the five brain regions in any group. In conclusion, chronic consumption of ethanol under the 'free-choice condition', which may best induce the drug-rewarding effect, leads to specific changes in the D3-receptor gene expression which were not seen after forced ethanol administration. Changes in D3 mRNA levels were, however, not a specific correlate of 'behavioral dependence', as they were also detected in rats not yet 'behaviorally dependent' (group b).

Impact of local donor and regionalization on a German transplantation center. UNI NRW. Universities of North Rhine Westfalia.

Optimal allocation of donor organs is an ongoing matter of debate. We report on the impact of the foundation of UNI NRW, a close transplant collaboration of seven university centers with the intention of improving donor organ allocation, on the heart transplant program in Münster. All donor organs retrieved were offered first to the patients within this region before going into the Eurotransplant (ET) Foundation pool. The heart transplant program data were prospectively (for 1997) and retrospectively (for 1996) analyzed with regard to donor organ availability and allocation. There was a slight decrease in the number of donor hearts offered and accepted within the UNI NRW region in 1997 as compared to in 1996. However, due to the significantly lower organ export rate, the number of heart transplantations performed in UNI NRW rose from 47 to 72 procedures. In Münster, only six donor organs (16%) were procured from outside UNI NRW in 1997, and these were, in part, due to special urgency requests. In conclusion, the institutionalization of UNI NRW within the framework of ET offers more flexibility, decreases total ischemic time, and may help to lower costs.