Serum adiponectin and resistin in rats under three isocaloric diets: The effect of sibutramine.

The aim of the study was to investigate the effects of (1) macronutrients on food intake, body composition and serum resistin and adiponectin and (2) sibutramine(S) on the above parameters in rats fed with isocaloric diets. Three groups of male Wistar rats (n=63) were fed with high fat diet (HFD), high carbohydrate diet (HCD) or high protein diet (HPD) for 13weeks. In the last 3weeks each group was divided into three subgroups and received S 5mg/kg or 10mg/kg, or vehicle. Body weight was measured weekly, gastrocnemius muscle, perirenal, retroperitoneal and epididymal fat were isolated, fat/lean ratio was calculated and serum adiponectin and resistin were assayed. S did not affect lean body mass in any group. HFD was associated with elevated fat/lean ratio regardless of S administration. S at 10mg/Kg decreased fat/lean ratio in the HCD and HPD and adiponectin in the HFD group. S did not affect resistin in any group. Adiponectin was paradoxically elevated in the HFDS10 compared to the HCD or HPD S10 groups. Resistin was lower in the HCD compared to the HPD and HFD groups. Results suggest a preferential effect of S on body fat. The detrimental effect of S on adiponectin can be attributed to its sympathomimetic properties. Adiponectin was paradoxically elevated in the HFD and resistin in the HPD group, results that require further investigation.

Mandibular bone density and calcium content affected by long-term anticonvulsant treatment in rats.

OBJECTIVE: Bone is perpetually absorbed and reformed, serving also to electrolyte homeostasis, mainly for calcium and phosphorus. Anticonvulsant medications are traditionally considered harmful to bone because of their interaction with the metabolism of vitamin D, due to hepatic enzyme induction. A study of the effect of anticonvulsant medications on mandibular bone quality was undertaken. MATERIALS AND METHODS: 24 Wistar rats in three groups received diphenylhydantoin or diazepam or placebo intraperitoneally (i.p.). Absolute bone weight, bone to body weight ratio, specific bone weight, absolute calcium concentration, calcium to mandibular bone weight ratio and mineral element concentration were examined after animal sacrifice, three months later. From the results it may be concluded that diazepam and diphenylhydantoin administration affect the mandibular bone density and calcium content in terms of absolute weight and specific weight. Mandibular calcium concentration was affected only by diphenhylhydantoin treatment.

Effect of isocaloric diets and sibutramine on food intake, body mass variation and serum TNF-alpha levels in rats.

BACKGROUND/AIM: The effect of isocaloric diets and sibutramine on dietary behaviour and TNF-alpha is poorly understood. The aim of the study was to investigate the effects of isocaloric diets and sibutramine on food intake, body mass variation and serum TNF-alpha in free-feeding rats. METHODS: Three groups of male Wistar rats (n = 63) were fed a high-fat diet, high-carbohydrate diet or high-protein diet for 13 weeks. In the last 3 weeks, each group was divided into 3 subgroups. Each subgroup received sibutramine 5 mg/kg, sibutramine 10 mg/kg or vehicle. Food intake was measured daily during the last week of the experiment; serum TNF-alpha was assayed and the body weight increasing rate was calculated. RESULTS: The high-fat diet was associated with increased food intake, a greater weight gain ratio and increased TNF-alpha levels. Sibutramine treatment did not affect the dietary behaviour of high-protein- or high-carbohydrate-fed rats, while it significantly attenuated the daily food intake and body weight gain rate in the high-fat group, at the dose of 10 mg/kg. TNF-alpha levels were not affected by sibutramine. CONCLUSIONS: High-fat feeding was associated with an increase in daily food intake, TNF-alpha levels and body weight gain rate, as well as with enhanced responsiveness to the anorectic effects of sibutramine. However, sibutramine did not affect TNF-alpha.

Development and optimization of a reversed-phase high-performance liquid chromatographic method for the determination of acetaminophen and its major metabolites in rabbit plasma and urine after a toxic dose.

A reversed-phase high-performance liquid chromatographic method with detection at 242 nm was developed, optimized and validated for the determination of acetaminophen (A) and its major metabolites glucuronide (AG) and sulfate (AS) conjugates in rabbit plasma and urine after a toxic dose. m-Aminophenol was used as internal standard (IS). A Hypersil BDS RP-C18 column (250 x 4.6 mm), 5 microm particle size, was equilibrated with a mobile phase composed of aqueous buffer solution of KH2PO4 0.05 M containing 1% CH3COOH (pH 6.5) and methanol (95:5, v/v). Its flow rate was 1.5 ml/min. Calibration curves of A, AG and AS were linear in the concentration ranges of 0.5-250, 1-200, 0.5-100 microg/ml in plasma and 1-200, 0.5-150, 0.5-100 microg/ml in urine matrix, respectively. Limits of detection and quantitation were calculated in all cases and extensive recovery studies were also performed. Intra-day relative standard deviation (R.S.D.) for A, AG and AS in plasma was less than 5, 4, 2% and in urine less than 4, 7, 4%, respectively, while the corresponding inter-day values were 7, 6, 4% and 5, 8, 6%, respectively.

Effects of mesulergine treatment on diet selection, brain serotonin (5-HT) and dopamine (DA) turnover in free feeding rats.

1. The effects of mesulergine, a 5-hydroxytryptamine (5-HT) receptor antagonist with dopamine (DA) agonistic properties, on rats diet selection over a seven day period and on 5-HT and DA turnover was studied. 2. Three groups of male Wistar rats were individually caged and ad libitum fed with a standard (SD) and 50% sweet carbohydrate enriched diet (CED). Food intake was measured daily 4 hrs and 24 hrs after i.p. injections of mesulergine (1 and 3 mg/kg) or vehicle. 5-HT and 5-HIAA in hypothalamus (Hy), Striatum (St) and hippocampus (Hi) as well as DA and DOPAC in (Hy) and (St) were assayed at the 8th day of the experiment. 3. There was a dose dependent increase of SD consumption 4 hrs after mesulergine treatment while the CED remained unchanged with total food intake dose dependently increased as a consequence. At 24 hrs measurements SD consumption was increased only for the dose of 1 mg/kg of mesulergine, while a dose dependent decrease of CED intake was observed. Total food intake was unchanged for the dose of 1 mg/kg and decreased with the dose of 3 mg/kg consequently. A dose dependent decrease of rats body weight was observed too. 4. A significant increase of 5-HIAA/5-HT ratio in (Hy) and (St) for the dose of 1 mg/kg and in (Hi) for the dose of 3 mg/kg with no changes of DA turnover were found. 5. The above data suggest a dual mode of action of mesulergine presented as a short term hyperphagia due to simultaneous antiserotonergic and dopaminergic activity and long-term hypophagia due to long-term agonistic effects of dopaminergic neurons.

Stress through handling for vaginal screening, serotonin, and ACTH response to ether.

The effect of duration of handling for vaginal smear screening on the adrenal weight and acute ACTH response to ether were examined in 4-day-cycling female rats, sacrificed at 97-103 days of age on diestrus-2 after evaluation of resistance to handling, thymus weight, and hypothalamic serotonin (5-HT) and 5-hydroxyindoleacetic acid (5-HIAA). Prolonged handling paralleled increased resistance (behavioral response) to handling and adrenal weight but was inversely related to thymus weight. The hypothalamic 5-HT, 5-HIAA, and 5-HIAA/5-HT ratio, compared to controls with similar conditions of handling, were not modified after 2.5 min of ether despite the ACTH rise. In ether-stressed rats, the ACTH response to ether was lower after prolonged handling compared to short handling paralleling decreased thymus weight. In contrast, 5-HT, 5-HIAA, and the 5-HIAA/5-HT ratio were higher, paralleling increased resistance and adrenal weight. The results suggest chronic activation of the hypothalamo-pituitary-adrenal axis with positive serotonergic involvement after prolonged handling and resistance during vaginal screening and a negative implication of this activation on the acute ACTH response to ether.

Behavioural changes on diet selection and serotonin (5-HT) turnover in rats under pizotifen treatment.

The effects of pizotifen on protein and carbohydrate self-selection in rats over a seven-day period, and on 5-HT turnover was studied. Four groups of male Wistar rats were individually caged and ad lib fed with a standard (SD) and (50%) carbohydrate-enriched diet (CED), sweet (diet group I) or not (diet group II). Food intake was measured daily 4 hr after IP injection of pizotifen (2.5 mg/kg) or vehicle. 5-HT and 5-HIAA in the hypothalamus (Hy), striatum (St) and hippocampus (Hi) were assayed on the 8th day of the experiment. Pizotifen increased the consumption of SD. The absolute intake of CED remained totally and daily unchanged, while the percentage proportion was reduced. Total food intake was increased by the drug which seemed to affect the proportion rather than the absolute amounts of carbohydrate and protein consumed. This effect was independent of the carbohydrate taste. There was a decrease of 5-HT levels in the Hi, while 5-HIAA/5-HT ratio was increased in the Hy and in the Hi of animals that consumed sweet carbohydrate. The above data suggest a role of pizotifen on 5-HT central metabolism and diet selection and support the view that changes of 5-HT metabolism in the Hy and Hi are responsible for protein selection and the regulation of SD/CED ratio, but they cannot explain drug's effect on total food intake.

Effect of enzyme induction on bioavailability of hetacillin in patients treated with anticonvulsants and chlorpromazine.

The effect of liver enzyme induction on bioavailability of hetacillin was studied in patients chronically treated with anticonvulsants or chlorpromazine. 24 chronic psychiatric patients classified according to their medication in two groups (anticonvulsants, chlorpromazine) and one group of 11 healthy volunteers, received an i.m. administration of 500 mg hetacillin. Serum levels of ampicillin derived from hetacillin in blood samples taken 2, 4 and 6 hours after the injection were measured and the half-life of the antibiotic was determined for each group. Urinary D-glucaric acid was considered the induction index. Correlation coefficients between the induction index and pharmacokinetic parameters of hetacillin were also determined. Anticonvulsants and chlorpromazine induced the liver microsomal enzymes as demonstrated by the increased D-glucaric acid excretion (P less than 0.001 - P less than 0.05). No statistically significant differences were found in serum levels and half-life of the antibiotic. Correlation coefficients suggest that enzyme induction and hetacillin bioavailability are not significantly related.

Drug-induced place preference in rats with 5,7-dihydroxytryptamine lesions of the nucleus accumbens.

The conditioned place preference (CPP) paradigm was used to determine a role for serotonin in the nucleus accumbens in the mediation of the rewarding properties of D-amphetamine morphine and diazepam. The effect of these drugs on CPP was examined in controls and in animals with 5,7-dihydroxytryptamine lesions of the nucleus accumbans. The results from control animals confirmed that D-amphetamine (1.5 mg/kg, i.p.), morphine (2.0 mg/kg, i.p.) and diazepam (1.0 mg/kg, i.p.) produced place preference for a distinctive environment that had previously been paired with injections of the drug. In animals with 80% reduction of 5-hydroxytryptamine content of the nucleus accumbens, D-amphetamine CPP was unchanged and morphine CPP was attenuated compared with controls. Diazepam CPP was not apparent in animals with the lesion. In separate experiments, characteristic behavioural effects of the drugs under study were examined in control and in animals with lesion. The results showed a tendency for increased amphetamine hyperlocomotion, enhanced morphine activity and analgesia and decreased diazepam anti-anxiety effect in animals with lesions. Thus, the 5,7-dihydroxytryptamine lesions of the nucleus accumbens differently influenced the CPP induced by the drugs studied and, with the exception of diazepam, the various behavioural effects elicited by each drug. The findings suggest that serotonin-containing neurones of the nucleus accumbens are a component of the neural circuitry that mediates the rewarding properties of morphine, probably of diazepam, but not of D-amphetamine.

Comparative study of the effects of some inducers with or without protein binding properties on bioavailability of isoxazolylpenicillins in rats.

The effect of various inducers with or without protein binding properties on serum levels and half life of Oxacillin, Cloxacillin and Dicloxacillin was studied. A total of 102 male rats classified in 3 "categories" according to the administered penicillin with 6 groups of rats in each of them were used. Each group was pretreated for 15 days with the following inducers: phenobarbital, diphenylhydantoin, diazepam, chlorpromazine and phenylbutazone. The control groups received saline. The d-glucaric acid concentration in the urine prior to and after the administration of inducers and the liver weight were taken as enzyme induction indices. The results showed a decrease of serum levels and half life of three penicillins with a negative correlation between urine d-glucaric acid and serum penicillin levels. Phenobarbital, diphenylhydantoin and chlorpromazine affected the 3 penicillins in the following statistically significant order: oxacillin, dicloxacillin, cloxacillin. Diazepam affected: cloxacillin, dicloxacillin, oxacillin, and phenylbutazone: dicloxacillin, cloxacillin and oxacillin. However all drugs finally produced a uniform effect on all 3 penicillins in the following decreasing order: phenobarbital (r = -0.910), diphenylhydantoin (r = -0.864), phenylbutazone (r = -0.851), chlorpromazine (r = -0.842) and diazepam (r = -0.821). For all inducers, the effect was most significant for oxacillin (r = -0.869), second most significant for dicloxacillin (r = -0.811) and finally for cloxacillin (r = -0.778). The results suggested an interaction of isoxazolylpenicillins and the above drugs.

Bioavailability of hetacillin in rats after liver enzyme induction by various inducers with or without protein binding properties.

The serum levels and the half life of ampicillin derived from Hetacillin after administration of the latter to a total of 61 rats classified in 7 groups were determined. Each of these groups was pre-treated for 15 days with the following inducers; phenobarbital, diphenylhydantoin, diazepam, chlorpromazine and phenylbutazone. The control group received saline. The d-glucaric acid concentration in the urine prior to and after the administration of inducers and the liver weight were taken as enzyme induction indices. Results showed a positive correlation between the indices of induction and the levels of ampicillin originating from hetacillin with a significant correlation coefficient between the serum levels of ampicillin and urine d-glucaric acid for all drugs studied. The different effect of the various drugs indicated that they could be classified into the following two groups: a) those that induced a significant increase of the levels and half life (t1/2) of ampicillin. The effect was significant in decreasing order for phenylbutazone (r = 0,990), diazepam (r = 0,990) and diphenylhydantoin (r = 0,753). b) those which initially resulted in a significant increase of the levels of ampicillin and thereafter in a decrease with a significant shortening of its t1/2 too. The effect was most significant for phenobarbital (r = 0,887) and less so for chlorpromazine (r = 0,800). Only for these two drugs was a significant and actually negative correlation observed between d-glucaric acid and t1/2 that is: phenobarbital (r = -0,967) chlorpromazine (r = -0,752). Results suggest an interaction of Hetacillin and the above inducers.

Influence of liver enzyme induction caused by diphenylhydantoin and diazepam on bone mass of rats.

25 albino rats of Wistar strain were used for the study. They were divided into three groups. Animals of the first group (n = 10) were treated with Diphenylhydantoin in a daily dose of 70 mg/kg for three months. Animals of the second group (n = 10) were treated with Diazepam in a daily dose of 10 mg/kg for 3 months. Animals of the third group (n = 5) were treated with 5% acacia oil for the same period of time. Food and water consumption together with motor activity were measured. As indices of enzyme induction, D-Glucaric acid and liver weight were determined. Also the following bone indices were determined: femur Ca/femur weight, femur Ca/body weight, humerus Ca/humerus weight, humerus Ca/body weight, femur specific weight, humerus specific weight. Statistically significant alterations of bone mass were found.