Effects of bumetanide on neurobehavioral function in children and adolescents with autism spectrum disorders.

In animal models of autism spectrum disorder (ASD), the NKCC1 chloride-importer inhibitor bumetanide restores physiological (Cl-)i levels, enhances GABAergic inhibition and attenuates electrical and behavioral symptoms of ASD. In an earlier phase 2 trial; bumetanide reduced the severity of ASD in children and adolescents (3-11 years old). Here we report the results of a multicenter phase 2B study primarily to assess dose/response and safety effects of bumetanide. Efficacy outcome measures included the Childhood Autism Rating Scale (CARS), the Social Responsive Scale (SRS) and the Clinical Global Impressions (CGI) Improvement scale (CGI-I). Eighty-eight patients with ASD spanning across the entire pediatric population (2-18 years old) were subdivided in four age groups and randomized to receive bumetanide (0.5, 1.0 or 2.0 mg twice daily) or placebo for 3 months. The mean CARS value was significantly improved in the completers group (P: 0.015). Also, 23 treated children had more than a six-point improvement in the CARS compared with only one placebo-treated individual. Bumetanide significantly improved CGI (P: 0.0043) and the SRS score by more than 10 points (P: 0.02). The most frequent adverse events were hypokalemia, increased urine elimination, loss of appetite, dehydration and asthenia. Hypokalemia occurred mainly at the beginning of the treatment at 1.0 and 2.0 mg twice-daily doses and improved gradually with oral potassium supplements. The frequency and incidence of adverse event were directly correlated with the dose of bumetanide. Therefore, bumetanide improves the core symptoms of ASD and presents a favorable benefit/risk ratio particularly at 1.0 mg twice daily.

Beneficial insulin-sensitizing and vascular effects of S15261 in the insulin-resistant JCR:LA-cp rat.

S15261, a compound developed for the oral treatment of type II diabetes, is cleaved by esterases to the fragments Y415 and S15511. The aim was to define the insulin-sensitizing effects of S15261, the cleavage products, and troglitazone and metformin in the JCR:LA-cp rat, an animal model of the obesity/insulin resistance syndrome that exhibits an associated vasculopathy and cardiovascular disease. Treatment of the animals from 8 to 12 weeks of age with S15261 or S15511 resulted in reductions in food intake and body weights, whereas Y415 had no effect. Troglitazone caused a small increase in food intake (P <.05). Treatment with S15261 or S15511 decreased plasma insulin levels in fed rats and prevented the postprandial peak in insulin levels in a meal tolerance test. Y415 had no effect on insulin levels. Troglitazone halved the insulin response to the test meal, but metformin gave no improvement. S15261 decreased the expression of phosphoenolpyruvate carboxykinase and glucose-6-phosphatase and stimulated the expression of acetyl-CoA carboxylase and acyl-CoA synthase. S15261 also reduced the expression of carnitine palmitoyltransferase I and hydroxymethyl-glutaryl-CoA synthase. S15261, but not troglitazone, reduced the exaggerated contractile response of mesenteric resistance vessels to norepinephrine, and increased the maximal nitric oxide-mediated relaxation. S15261, through S15511, increased insulin sensitivity, decreased insulin levels, and reduced the vasculopathy of the JCR:LA-cp rat. S15261 may thus offer effective treatment for the insulin resistance syndrome and its associated vascular complications.

Pulmonary alveolar proteinosis in Israel: ethnic clustering.

BACKGROUND: Pulmonary alveolar proteinosis is a rare disease in which a surfactant-like phospholipid-rich protein accumulates in the lungs. The disease is amenable to effective therapy by total lung lavage. OBJECTIVES: To investigate the prevalence, ethnic distribution and course of PAP in Israel. METHODS: A countrywide survey was conducted during which pulmonologists were questioned about patients with PAP. The patients were examined and their charts, radiological images, pathological slides and physiological data were reviewed. RESULTS: The survey yielded 15 patients (8 females) during the period 1976-98 (14 in the last decade), giving a prevalence of 3.7 x 10(6) and an incidence of 0.36 x 10(6)/year. Mean age of the patients was 33 +/- 13 years (range 0.5-46 years). Seven patients were North African (two were siblings), four were from Iraq and two were Arabs; there was only one Ashkenazi Jew (a child). Symptoms at the onset were dyspnea and chest pain. Spontaneous remission occurred in at least 3 patients, and 10 patients required 1-4 bronchoalveolar lavage treatments. The subjective and physiological response was favorable, but there was less consistent radiological improvement. CONCLUSION: The prevalence of PAP in Israel is approximately 3.7 x 10(6). Most cases occurred in Jews who had immigrated from North Africa or Iraq, and two were siblings. The prevalence among the Arab population appears to be similar. This clustering suggests the existence of a genetic predisposition. The course of the disease appears to be similar to that reported elsewhere.

Effect of gliclazide treatment on insulin secretion and beta-cell mass in non-insulin dependent diabetic Goto-Kakisaki rats.

The Goto-Kakisaki rat is a genetic non-overweight model of non-insulin-dependent diabetes mellitus. Adult Goto-Kakisaki rats exhibit a mild basal hyperglycaemia (11 mmol/l) with impaired glucose tolerance, elevated basal plasma insulin level, a failure of insulin release in response to glucose together with a 50% depletion of the total pancreatic beta-cell mass and insulin stores. We have examined the effects of long-term (4 weeks) gliclazide treatment on the severity of diabetes in adult male Goto-Kakisaki rats (10-12 weeks of age). Gliclazide was administered orally (10 mg/kg per day). Gliclazide-treated Goto-Kakisaki rats were evaluated against Wistar and untreated Goto-Kakisaki rats. In the gliclazide-treated Goto-Kakisaki rats, basal plasma glucose levels declined progressively reaching 8 mmol/l as a mean at the end of treatment, and their basal insulin levels decreased to values similar to those in non-diabetic Wistar rats. Despite their total pancreatic beta-cell remaining unaffected, their pancreatic insulin stores were twice increased, with a similar improvement of the insulin content per individual beta-cell. Furthermore, the glucose-stimulated insulin release as evaluated in vivo during an intravenous glucose tolerance-test was significantly improved (twice increased) in the gliclazide-treated Goto-Kakisaki rats. This was correlated with a modest but significant enhancement of the early phase of insulin release in vitro (isolated perfused pancreas), in response to glucose. However, the overall insulin response in vitro remained clearly defective with no reappearance of the late phase of insulin release. The in vitro response to arginine (which was basically amplified in the Goto-Kakisaki model) or to gliclazide were kept unchanged after the gliclazide treatment. In conclusion, chronic gliclazide does not exert any beta-cytotrophic effect, but improves beta-cell function in the adult Goto-Kakisaki rat as far as it lowers basal insulin release, increases beta-cell insulin stores, and increases the glucose-induced insulin release.

Metabolic and anti-atherogenic effects of long-term benfluorex in dyslipidemic insulin-resistant sand rats (Psammomys obesus).

Benfluorex is a clinical lipid-lowering agent with antihyperglycemic properties. The effect of long-term oral treatment (10 mg/kg/day for 7.5 months) on carbohydrate and lipid metabolism and aortic morphology was investigated in 24 insulin-resistant sand rats receiving a standard laboratory diet supplemented with cholesterol (2%). Untreated controls (n=34) developed impaired glucose tolerance, hyperinsulinemia, hypertriglyceridemia, hypercholesterolemia and elevated plasma LDL- and VLDL-cholesterol, positively correlated with the proportion of the thoracic aorta displaying oil red O-positive atherosclerosis; ultrastructural examination showed intimal lipid deposits, foam cells, polymorph infiltrates and fibrosis. Benfluorex-treated animals showed significant decreases in glucose intolerance, hyperinsulinemia, hypertriglyceridemia, hypercholesterolemia, and plasma LDL- and VLDL-cholesterol, with no evidence of aortic atheroma. The metabolic benefits of benfluorex may protect against the long-term development of atherosclerosis in the insulin-resistant dyslipidemic syndrome.

Acute and chronic effects of dexfenfluramine on the porcine pulmonary artery.

1. Studies were designed to investigate the responses of isolated pulmonary arteries from control pigs or pigs chronically treated with dexfenfluramine (7.2 mg/kg per day orally for 4 weeks). 2. Rings with and without endothelium were suspended in organ chambers for recording of isometric tension. 3. Dexfenfluramine (10[-9] to 10[-6] M) did not affect vascular tone, but at higher concentrations caused contractions that were not affected by indomethacin, methiothepin, the presence of endothelium or by the chronic treatment. 4. Chronic treatment augmented the endothelium-dependent relaxations to serotonin and aggregating platelets but not those to adenosine diphosphate. It did not affect the contraction or rings without endothelium evoked by platelets, nor the relaxation to SIN-1, a nitric oxide donor. The maximal contraction to endothelin-1 (but not that of norepinephrine) was impaired in treated pigs. 5. These results show that dexfenfluramine causes contraction of isolated porcine pulmonary arteries only at concentrations higher than 3 x 10(-6) M, and that chronic treatment with dexfenfluramine potentiates the endothelium-dependent relaxations to serotonin and aggregating platelets in the porcine pulmonary artery without affecting their direct effect on the smooth muscle.

Vascular action of the hypoglycaemic agent gliclazide in diabetic rabbits.

ATP-dependent potassium channel blockers used as hypoglycaemic agents may have effects on vascular disease in diabetes mellitus beyond their effect on blood glucose control. This study was designed to determine the effects of treatment with gliclazide on the isolated abdominal aorta of diabetic rabbits in which endothelium-dependent relaxation is impaired by a mechanism involving oxygen-derived free radicals. After induction of diabetes with alloxan, there was no effect of gliclazide (10 mg x kg(-1) day(-1) orally) on blood glucose or insulin levels over a 6 week period. Hence, this permitted an examination of the vascular effects of gliclazide in diabetic rabbits exclusive of metabolic effects. Acetylcholine- and nitric oxide-induced relaxation in aortae from rabbits treated with or without gliclazide were measured in the absence or presence of the nitric oxide synthase inhibitor, N(G)-nitro-L-arginine (L-NAME). Diabetes was associated with significant impairment of acetylcholine-induced endothelium-dependent relaxation of the abdominal aorta which was not significant in diabetic rabbits treated with gliclazide in vivo. Aortae from diabetic rabbits studied in the presence of L-NAME showed an exaggerated contraction to acetylcholine which was prevented in rabbits treated with gliclazide. Gliclazide treatment did not affect the response to acetylcholine of normal rabbit aorta, and gliclazide when added in vitro had no effect on the response of diabetic rabbit aorta, suggesting that the effect of gliclazide was specific to the abnormality arising with diabetes and was not due to an acute effect of the drug. These data indicate that gliclazide, aside from either a direct antioxidant action or an effect on insulin or glucose levels, may ameliorate diabetic endothelial cell dysfunction.

Acute and chronic effects of dexfenfluramine on the porcine coronary artery.

Experiments were designed to verify whether or not acute or chronic exposure to dexfenfluramine favors the occurrence of coronary vasospasm in vivo or in vitro. Rings of left anterior and left circumflex porcine coronary artery, with and without endothelium, were studied in conventional organ chambers for the measurement of isometric force. The donor pigs were divided into two groups: controls and animals fed for 4 weeks with dexfenfluramine. In each group, one-half of the animals underwent balloon denudation of the left anterior descending coronary artery at the beginning of the study. Coronary angiography was performed at the time of denudation and, in all animals, during the 3rd week of the study. Acutely, dexfenfluramine at concentrations higher than 10(-5) M caused contractions which were blunted by the presence of the endothelium and inhibited by indomethacin (an inhibitor of cyclooxygenase). Chronic treatment with dexfenfluramine did not affect coronary diameter and did not alter the response to intracoronary infusion of serotonin. Chronic treatment with dexfenfluramine reduced the contractions of rings without endothelium to serotonin, but not those to norepinephrine or endothelin. It did not affect endothelium-dependent relaxations in the absence or presence of pertussis toxin to serotonin, UK14304 (alpha-2 adrenergic agonist), adenosine diphosphate or aggregating platelets. Chronic treatment with dexfenfluramine did not modify relaxations of rings without endothelium to SIN-1 (nitric oxide donor; the active metabolite of molsidomine) or adenosine diphosphate. These findings do not support the hypothesis that acute or chronic exposure to dexfenfluramine favors the occurrence of coronary vasospasm.

Research prospects with benfluorex.

The recent recognition that insulin resistance is associated with a number of risk factors for atherosclerotic cardiovascular disease has increased the interest in agents that are able to improve insulin sensitivity. The capacity of benfluorex (Médiator) to enhance insulin action has led to much speculation regarding its mechanism of action. Chronic benfluorex treatment, in a variety of genetic and dietary animal models of diabetes and insulin resistance, has been shown to diminish, circulating insulin levels and to decrease blood glucose, triglycerides, and cholesterol concentrations. From these studies, it is possible to postulate a multifactorial mode of action of this drug that involves three independent but interactive processes: (1) a direct effect on insulin target tissues, mediated by mechanisms distal to the binding of insulin to its receptor, (2) modulation of the glucoregulatory hormone balance, including a diminution in both adrenal and sympathetic tone, leading to improved hepatic sensitivity to insulin, and (3) reduced hepatic and muscle lipid availability, leading to improved glucose utilization in skeletal muscle. The multiplicity of the neuroendocrine and biochemical effects of benfluorex cannot be explained by a single cellular or molecular action. It has been suggested that insulin sensitizers may act on key molecules involved in the sequence of biochemical events involving the insulin signal transduction process. The identification of these molecular targets and the determination of their relative importance in the treatment of type II diabetes remains to be established and constitutes the main subject of ongoing research with benfluorex.

Euglycaemic and biological activities of novel thiazolidine-2,4-dione derivatives.

A new series of thiazolidine-2,4-dione derivatives was obtained by incorporating one or the other of the two carbons of the central chain into different rings. These compounds lower blood glucose levels in the genetically obese and insulin-resistant ob/ob mouse. Moreover, they decreased insulin and triglyceride levels in the Zucker fa/fa rat. Incorporation of the left hand carbon of the chain afforded compounds among which pyrrolidino derivatives 5, 9 and 13 were the most potent. The same carbon atom was used to elaborate different types of rings (benzocyclobutane, benzodioxane), giving rise to compounds 14 and 19 with moderate to good activity. Finally, cyclization using the right hand carbon of the chain gave rise to highly potent benzofurane 24.

[Intravenous glucose tolerance test in the functional exploration of segmental pancreatic autografts in the dog]. / L'épreuve d'hyperglycémie provoquée par voie intraveineuses dans l'exploration fonctionnelle de l'autogreffe pancréatique segmentaire chez le chien.

The aim of the present study is based on the comparison of intravenous tolerance testing before and after segmental pancreas autotransplantation in the dog. The results show that such testing must take in account the "glucose diffusion space", using the same glucose load in order to avoid the bias related to the post-operative loss of body weight.

S15261, a new compound for the treatment of the insulin resistance syndrome.

A new oral agent, S15261 (the L-isomer of 3-[2-[2-[4-[2-[alpha-fluorenyl acetyl amino ethyl] benzoyloxy] ethyl amino] 1-methoxy ethyl] trifluoromethyl-benzene), has been developed for the treatment of the so-called "insulin resistance syndrome". In obese, insulin-resistant ageing Sprague-Dawley rats, chronic treatment with S15261 (0.5-2.5 mg.kg-1.day-1 twice per day, for 14 days) resulted in dose-dependent decreases in plasma insulin (43%), and triglyceride levels (36%), and in an increase of the glucose disposal rate during an intravenous glucose tolerance test (IVGTT) (48.5%). An increase in peripheral insulin sensitivity produced by S15261 was revealed by the glucose clamp technique. Thus, the glucose infusion rate was increased by 20% whilst steady-state insulin levels decreased by 15%. At the higher doses S15261 led to a decrease in body weight (3%), plasma glucose (13%) and blood pressure (8 mm Hg) in mildly hypertensive animals. At the doses used to achieve these results, the compound has no hypoglycaemic activity in normoglycaemic animals. Acute administration of S15261 directly into the portal vein provoked a marked increase in glucose disappearance rate during an intravenous glucose tolerance test (60%) and also in the pancreatic response to the glucose challenge. Thus, acute administration of the compound has a direct effect on glucose metabolism. These data suggest that S15261 could be a useful agent for the treatment of the insulin resistance syndrome.

[Functional and morphological study of the pancreatic graft after segmental heterotopic autotransplantation in dogs]. / Etude fonctionnelle et morphologique du greffon pancréatique après autotransplantation segmentaire hétérotopique chez le chien.

Segmentary heterotopic autotransplantation of the body and tail of the pancreas, with visceral exocrine drivation and splenosplenic arterio-venous fistulation, was performed in 12 Beagle dogs. An in vivo glucose tolerance test was conducted before the transplantation and 21 days after the operation. There was a significant decrease in the k coefficient (2.884 +/- 0.234 before and 1.878 +/- 0.128 after transplantation) due to reduced peripheral glucose uptake after transplantation (p < 0.001). Overall glucose-stimulated insulin production was decreased after < 0.001). Two populations were identified retrospectively: in 7 dogs insulin response was satisfactory after transplantation (insulin production > or = 50% pretransplantation level) and in 5 the response was below 50%. Glucose tolerance was tested in vitro in the isolated perfused pancreas transplant in 9 dogs, 30 days after the transplantation. Secretory response was assessed according to the early peak of insulin secretion after glucose stimulation. The patterns of insulin secretion were not different before glucose stimulation but became statistically different after stimulation (p < 0.001). In 5 dogs, the response to in vitro glucose stimulation showed an early peak in insulin secretion and in 4 dogs the insulin response came late with no early peak. On histological examination normal (or subnormal) pancreas grafts and grafts in which extensive sclerosis impaired function could be distinguished. There was a significant correlation between the quality of function and the histology, suggesting that containing post-operative sclerosis to a minimum is an important factor in human transplantations.

[Surgical imperatives in pancreatic transplantation in dogs]. / Impératifs chirurgicaux de la transplantation pancréatique chez le chien.

The authors report their experience of segmental pancreatic autotransplantation in dogs. The most reliable model seems to be: segmental heterotopic pancreas transplantation with bladder diversion of the exocrine secretion and spleno-splenic arterio-venous fistula allowing monitoring of the exocrine secretion and a reduction in the thrombosis rate.

Insulin release from isolated islets of Langerhans after segmental-pancreas autotransplantation in dogs.

Pancreas transplantation is warranted essentially by the quality of glucose regulation. Although the fasting blood glucose is invariably normal, this may not be the case during glucose load tests. The purpose of this study was to examine dysregulation within the isolated islet originating from a segmental-pancreas autograft in the dog. Results show an increased basal insulin secretion by the graft islets in static incubation compared with that of islets originating from the head of the pancreas and left in situ. This abnormal secretion may be accounted for by various factors intervening within the graft or the isolated islet, thus suggesting a possible improvement in the surgical model.

Essential fatty acid interconversion during gestation in the rat.

The synthesis of arachidonic acid has been investigated in fetal and pregnant rat liver microsomes in the course of the gestation. The delta 5-desaturase activity decreased 2-3 times in rat liver between the 19th and 22nd day of the pregnancy. During this period the delta 5-desaturate activity increased 3-fold in the fetal liver, exceeding the activity of the maternal liver. In contrast, the activity of the fetal delta 6-desaturase was in the same range as in pregnant rat liver and the liver of control animals and did not change between these two stages of the gestation. The elongation rate of linoleic acid in fetal liver was 2-3 times lower than in maternal liver but this increased during the pregnancy. The fatty acid activate rate was always higher than the activity of the desaturases. At the 19th day, the activity of the delta 5-desaturase was apparently the rate limiting step of arachidonic acid synthesis in fetal liver. We did not find any delta 5- and delta 6-desaturase activities or linoleic acid elongation in the placenta microsomes.

Essential fatty acids interconversion in the human fetal liver.

In order to study the role played by the fetoplacental unit in providing the human fetus with arachidonic acid, delta 5- and delta 6-desaturase activities were studied in microsomes from human fetal liver and placenta after 18 and 22 weeks of gestation. We evidenced for the first time delta 5- and delta 6-desaturase activities in fetal liver microsomes. As in adult liver, delta 6-desaturation is the rate-limiting step of arachidonic acid synthesis. No activity was found in the placenta. Arachidonic acid concentrations were higher in fetal serum than in maternal serum while the opposite was observed for linoleic acid. The fetal liver microsomal content in arachidonic acid was low. Taken together the data suggest that arachidonic acid is supplied to the fetus through a preferential transfer across the placenta.