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Biomedical subjects

G Chaudhuri

Publications and source records attributed to G Chaudhuri.

At least 55 records · Page 3Linked to original sources

Cloning and sequencing of a cDNA encoding a novel member of the human brain GABA/noradrenaline neurotransmitter transporter family.

Screening of a human hippocampal cDNA library with a rat creatine transporter cDNA-specific probe revealed two types of clones. One identical to the published creatine transporter cDNA sequence (CRT1) and another (CRT2) with four segments of oligodeoxyribonucleotide substitution or addition in different sites of the CRT1 coding sequence. Translation of the CRT2 coding sequence reveals a new protein with regions of perfect homology with the CRT1 amino-acid sequence.

Amino Acid Sequence↗

The protective effect of tetrahydrobiopterin on the nitric oxide-mediated inhibition of purified nitric oxide synthase.

The nitric oxide synthases (NOS) are a class of enzymes responsible for the generation of NO via an oxygen and NADPH dependent oxidation of the amino acid arginine. These enzymes are ironheme proteins which contain FAD and FMN and, enigmatically, require tetrahydrobiopterin (BH4). NOS has recently been shown to be subject to inhibition by its product, NO. Preliminary data by us indicate that a possible role for BH4 is to prevent and/or reverse the NO-mediated inhibition of NOS. The objective of this study was to elucidate the mechanism by which BH4 protects NOS against NO inhibition. Protection of NOS from NO inhibition was observed by both BH4 and the BH4 regeneration system, dihydropteridine reductase (DHPR)/NADH. NO, rather than an oxidation product, appears to be the inhibitory species. Protection by BH4 is not likely due to a simple chemical reaction between BH4 and NO or its oxidation product, NO2. The results are consistent with a protective mechanism by which BH4 may act as a nonstoichiometric reducing agent for a redox active enzyme component, such as the ironheme, to prevent NO ligation.

Amino Acid Oxidoreductases↗

Nitric oxide donors induce extrusion of cyclic GMP from isolated human blood platelets by a mechanism which may be modulated by prostaglandins.

In the presence of 3-isobutyl-methylxanthine (IBMX), induction of cyclic 3',5'-guanosine monophosphate (GMP) production in human washed platelets (HWP) by nitric oxide donors (NOD) is followed by its accumulation in the surrounding medium in a time- and concentration-dependent manner. Thirty minutes incubation of HWP with 3-morpholino-sydonimine (SIN-1, 10 microM) at 37 degrees C resulted in a 4.6-fold increase of cyclic GMP in platelets, whereas in the extracellular medium the increase was 17.6-fold. Similar results were obtained when other NOD such as S-nitroso-N-acetylpenicyllamine (SNAP) and 3-(2-methoxy-5-chlorophenyl)oxatriazol-5-imine (GEA 3184) and the selective phosphodiesterase inhibitor, zaprinast (M&B 22948, 10 microM), were used. Probenecid (1-300 microM), an inhibitor of organic anion transport, or ouabain (1-300 microM), an inhibitor of Na+/K+ adenine triphosphate (ATP)-ase had no effect on cyclic GMP production or extrusion after stimulation with SIN-1. Significantly prostaglandin A1 (PGA1) and prostaglandin D2 (PGD2) inhibited the efflux of cyclic GMP from platelets induced by SNAP (10 microM) in a concentration-dependent fashion, with an IC50 of 63 +/- 16 and 143 +/- 17 microM, respectively. These studies suggest that the extrusion of cyclic GMP from human platelets after activation of soluble guanylate cyclase by NOD may contribute to the control of cyclic GMP levels in platelets with potential physiological and therapeutic consequences.

1-Methyl-3-isobutylxanthine↗

Gender differences in atherosclerosis: possible role of nitric oxide.

The mechanism by which women in the reproductive age group are protected from developing coronary heart disease (CHD) as compared with men of similar age is not known. To elucidate whether there is a gender difference in the rate of atherosclerosis formation, we investigated the rate of development of atherosclerosis in both male and female rabbits fed an identical diet consisting of 2% cholesterol for 10 and 15 weeks. The extent of atherosclerosis was correlated with the amount of basal and stimulated release of nitric oxide (NO) from endothelium-intact aortic rings obtained from these animals. Under identical dietary conditions, the female rabbits fed a high cholesterol diet (HCD) for 10 weeks developed very little atherosclerosis (10% surface involvement) as compared with male rabbits (42% surface involvement). However, no significant gender differences in atherosclerosis were observed after 15 weeks of the HCD. The serum cholesterol, high and low density lipoprotein (HDL and LDL) cholesterol were similar in animals fed the HCD for 10 and 15 weeks. The basal release of NO from endothelium-intact aortic rings was significantly greater in control females as compared with males. The magnitude of endothelium-dependent relaxation of aortic rings obtained from both male and female rabbits fed the HCD were impaired to a similar extent, and this impairment correlated with the duration of hyperlipidemia but not with the extent of atherosclerosis. The arginine content of aortic rings were not different between males (257 +/- 52 nmol/g wet weight) and females (345 +/- 62 nmol/g wet weight) or between control and hyperlipidemic groups (males 312 +/- 69; females 301 +/- 65 nmol/g wet weight). Although the precise mechanism for the slower rate of development of atherosclerosis in the female rabbits as compared with males is not clear, the greater basal release of NO in females before they were fed a hyperlipidemic diet, as well as other factors, may be involved. The impairment of endothelium-dependent relaxation in hyperlipidemic animals is not due to a decrease in the availability of arginine, the substrate for NO.

Acetylcholine↗

The role of nitric oxide in the altered vascular reactivity of pregnancy in the rat.

1. Pregnancy is characterized by a decrease in systemic vascular resistance and a blunting of the angiotensin II (AII) pressor response. We studied the role of nitric oxide (NO) and prostanoids in these vascular changes of pregnancy in anaesthesized, ganglion blocked non-pregnant and pregnant rats. 2. Inhibition of NO synthesis with NG-nitro-L-arginine methyl ester (L-NAME) led to an increase in mean arterial pressure (MAP) which was of a significantly greater magnitude in pregnant rats in late gestation than in non-pregnant rats, or rats in mid-gestation. 3. The pressor response to varying doses of AII was attenuated during late pregnancy, and this attenuation was partially reversed by L-NAME. 4. The pressor response to varying doses of a vasoconstrictor, phenylephrine (PE), was also attenuated in late pregnancy. However, this attenuation was not reversed by L-NAME. 5. Inhibition of prostanoid biosynthesis with meclofenamate did not alter basal MAP, nor the pressor response to varying doses of AII or PE in pregnant and non-pregnant animals. 6. It is concluded that (a) increased NO synthesis occurs during late gestation and contributes both to the decrease in systemic vascular resistance, as well as the blunting of the pressor response to AII during pregnancy, and (b) prostaglandins are not important in the maintenance of basal vascular tone, or the blunting of the pressor response to AII during pregnancy.

Angiotensin II↗

Haemostatic mechanism in the endometrium: role of cyclo-oxygenase products and coagulation factors.

1. The primary mechanism of haemostasis in the endometrium of rat was studied and results were compared to that in the mesenteric artery. 2. The bleeding time of the rat endometrium as assessed by haemoglobin output was significantly decreased after pretreatment of the animals with either indomethacin (5 mg kg-1, i.v.) or meclofenamate (3 mg kg-1, i.v.) whereas the bleeding time was significantly increased in the rat mesenteric artery. 3. The bleeding time of the rat endometrium was unchanged from control values following treatment with prostacyclin (0.5 microgram kg-1 min-1, i.v.) or 1-benzylimidazole (50 mg kg-1, i.v.) whereas the bleeding times were increased in the rat mesentric artery. 4. Administration of heparin (100 units kg-1) increased the bleeding time in the rat mesenteric artery but had no effect on the bleeding time of the endometrium. 5. Superfusion of the endometrium with 16, 16-dimethyl PGE2 (1 microgram ml-1) a vasodilator, increased the bleeding time of the endometrium but superfusion of PGE2 over the mesenteric artery did not affect the bleeding time from this site. 6. Histological studies of the mesenteric artery and the endometrium following haemostatis revealed that the haemostatic plug in the mesenteric artery was mainly composed of platelets and fibrin whereas in the endometrium it was mainly composed of fibrin. 7. These findings suggest that haemostasis in the endometrium may be mediated by the vascular tone and fibrin whereas formation of the platelet plug may be primary mechanism for haemostasis in the mesenteric artery.

Animals↗

Inhibition of constitutive and inducible nitric oxide synthase: potential selective inhibition.

Nitric oxide (NO) is a molecule that has been shown to be involved in a diverse array of physiological events. A variety of disease states and disorders are, in fact, due to either an over- or an underproduction of NO. As a result of the ubiquity and diversity of NO-mediated phenomenon, pharmacological manipulation is difficult. NO biosynthesis is the result of an oxidation of a terminal nitrogen on the amino acid arginine by a class of enzymes generally referred to as the nitric oxide synthases (NOSs). Since the various isoforms of NOS are distributed in cells and tissues according to their function, there is the possibility that manipulation of NO levels can be accomplished by designing specific pharmacological agents targeted at a single NOS isoform. Thus, this review discusses general inhibition of the NOSs by a variety of agents and then focuses on the possibility of developing agents for specific isoform inhibition.

Amino Acid Oxidoreductases↗

H2 gas clearance technique for separating rat uterine blood flow into endometrial and myometrial components.

The H2 gas clearance technique was employed to measure uterine blood flow (UBF) in ovariectomized rats. A needle-type platinum electrode (125 microns diam) was inserted into the rat uterine wall to measure the tissue blood flow surrounding the electrode. The electrode can be placed in individual layers of the uterus to measure the endometrial blood flow (EBF) or the myometrial blood flow (MBF). By use of this technique, baseline EBF and MBF were 37.8 +/- 3.53 (n = 21) and 47.2 +/- 4.56 (n = 5) ml.min-1.100 g-1, respectively, with an EBF/MBF ratio of 0.8. Intravenous bolus injection of 17 beta-estradiol (1 microgram/kg) induced a significant increase in UBF. Phenylephrine, an alpha-adrenergic receptor agonist, reduced UBF. In some animals, a second platinum electrode was used to measure gastric mucosal blood flow simultaneously with UBF. While 17 beta-estradiol selectively increased UBF, pentagastrin selectively increased gastric mucosal blood flow. To further validate the baseline UBF distribution between endometrial and myometrial layers, iodo[14C]antipyrine autoradiography was employed. With the iodo[14C]antipyrine technique, the EBF/MBF ratio was 0.91 +/- 0.07 (n = 5), which is similar to that obtained with the H2 gas clearance technique.

Animals↗

Nitric oxide is an important mediator for tumoricidal activity in vivo.

When cultured in vitro, peritoneal macrophages, obtained from mice previously inoculated with bacillus Calmette-Guérin, release nitric oxide, which is cytostatic and/or cytolytic for tumor cells. However, it is not known whether nitric oxide has antitumor effects in vivo. Here we demonstrate that nitric oxide is an important mediator of host resistance to syngeneic and xenogeneic ovarian tumor grafts in C3HeB/FeJ mice. A murine ovarian teratocarcinoma cell line, utilized to study the mechanism of bacillus Calmette-Guérin-induced host resistance to a syngeneic ovarian tumor, proliferated when transplanted intraperitoneally. Marked tumoricidal activity was observed, however, when these murine ovarian teratocarcinoma cells were transplanted 8 days after intraperitoneal bacillus Calmette-Guérin inoculation. In studies related to xenogeneic ovarian tumor grafts, tumoricidal activity was observed after intraperitoneal transplantation of a human epithelial ovarian cancer cell line, NIH:OVCAR-3. This cell line proliferates only in athymic nude (immunologically incompetent) mice. In both sets of experiments, tumoricidal activity was reduced by inhibition of nitric oxide synthesis. These results demonstrate the tumoricidal action of nitric oxide in vivo.

Animals↗

Estradiol does not influence myocardial superoxide dismutase activity in rabbits.

The influence of sex steroids on myocardial superoxide dismutase (SOD) activity was evaluated in rabbits. No sex differences in SOD activity were observed in male and female rabbits. Castration did not lead to any changes in SOD activity in either sex. Graded doses of estradiol that produced graded increases in uterotropic effect failed to produce any change in myocardial SOD activity. Results indicate that the beneficial effect of estrogen on cardiovascular disease and the sex differences in incidence of cardiovascular morbidity cannot be explained on the basis of a direct action of estradiol on myocardial SOD activity.

Animals↗

NO is more important than PGI2 in maintaining low vascular tone in feto-placental vessels.

The endothelial cells of the human umbilical artery and vein release the vasodilators prostacyclin [prostaglandin (PG) I2] and nitric oxide (NO). However, the role of these two substances in the maintenance of vasodilator tone in the feto-placental circulation is not known. Studies were therefore undertaken to compare the relative release of PGI2 and NO from perfused segments (10 cm) of endothelium-intact human umbilical artery (HUA) and vein (HUV) utilizing the cascade bioassay. The endothelium-denuded bovine pulmonary arterial strip was used as the detector tissue because this tissue relaxes equally to various concentrations of PGI2 and S-nitroso-N-acetylpenicillimine (SNAP), which acts by releasing NO. The basal release of NO from the HUA was approximately five times greater than that of PGI2. After stimulation with A-23187, the release of NO from HUV was five to six times greater, and from the HUA, the release was three times greater compared with the PGI2. SNAP was significantly more potent compared with PGI2 in relaxing endothelium-denuded rings of human umbilical and chorionic plate arteries in vitro. These studies suggest that NO is more important than PGI2 for maintenance of low vascular tone in feto-placental vessels, because there is a greater release of NO from the HUA and HUV, and NO is more potent in relaxing endothelium-denuded feto-placental vessels in vitro relative to PGI2.

Animals↗

Basal release of nitric oxide from aortic rings is greater in female rabbits than in male rabbits: implications for atherosclerosis.

Estradiol is known to exert a protective effect against the development of atherosclerosis, but the mechanism of this hormonal action is unknown. One of the early events in the development of atherosclerosis is the adhesion of macrophages to endothelial cells, and nitric oxide (NO) inhibits this process. We show that basal release of NO is greater with endothelium-intact aortic rings from female rabbits than those from males. Oophorectomy diminishes both circulating estradiol concentration and basal release of NO to levels seen in male rabbits. These data establish that basal NO release from endothelium-intact aortic rings depends on circulating estradiol concentration and offer an explanation for the protective effect of estradiol against the development of atherosclerosis.

Acetylcholine↗

Chemical oxidation of N-hydroxyguanidine compounds. Release of nitric oxide, nitroxyl and possible relationship to the mechanism of biological nitric oxide generation.

N omega-Hydroxy-L-arginine was found to cause vasodilation in arginine-depleted rabbit aorta. It is, therefore, likely to be a biosynthetic intermediate in the conversion of arginine to nitric oxide in this tissue. N-Hydroxyalkylguanidine compounds, including N omega-hydroxy-L-arginine were oxidized with various oxidizing agents and examined for their ability to release nitric oxide. All oxidizing agents tested were capable of oxidizing the N-hydroxyguanidine function but only lead tetra-acetate (Pb(OAc)4) and potassium ferricyanide/hydrogen peroxide (K3FeCN6/H2O2) were capable of generating significant amounts of nitric oxide. Oxidation with K3FeCN6, lead oxide (PbO2) and silver carbonate (Ag2CO3) resulted instead in the release of nitrous oxide (N2O) presumably through the initial release of nitroxyl (HNO).

Animals↗

Nitric oxide and prostacyclin inhibit fetal platelet aggregation: a response similar to that observed in adults.

OBJECTIVE: We evaluated the relative importance of two endothelium-derived substances, prostacyclin and nitric oxide, in their ability to inhibit aggregation of fetal and maternal platelets. STUDY DESIGN: The effects of various concentrations of prostacyclin and S-nitroso-N-acetylpenicillamine (which releases nitric oxide) on platelet aggregation were studied by means of platelet-rich plasma from at least five to six subjects per group. Fetal blood was collected from umbilical vein at delivery. Maternal venous blood was collected within 4 hours of delivery. Platelet aggregation was monitored with a platelet aggregation profiler. Adenosine diphosphate was used as the aggregating agent. Statistical differences between means were evaluated with two-way analysis of variance or Student t test. RESULTS: Prostacyclin and S-nitroso-N-acetylpenicillamine inhibited aggregation of fetal and maternal platelets, but prostacyclin was more potent. Fetal platelets were more sensitive than maternal platelets to prostacyclin and S-nitroso-N-acetylpenicillamine. CONCLUSION: Prostacyclin appears to be more important in preventing aggregation of platelets in the feto placental circulation.

Adenosine Diphosphate↗

Prostaglandins in the ovary and fallopian tube.

More than 20 years following the recognition of a possible role for eicosanoids in ovarian function a physiological role for prostaglandins and/or leukotrienes in human ovulation, corpus luteum function and tubal motility remains to be demonstrated. With respect to ovarian function, the well-characterized preovulatory rise in eicosanoid production in animal species and humans, in conjunction with the large body of experimental evidence employing inhibitors of prostaglandin synthesis and replacement of individual prostaglandins, has provided strong evidence for a role in follicular rupture independent of other LH-mediated ovulatory events. The possible mechanism of prostaglandin-induced follicle rupture may involve stimulation of proteolytic activity via substances such as plasmin and PA; however, this is controversial. A role for prostaglandins in ovarian luteal function is well established in laboratory animals and large ruminant species, where PGF2 alpha derived from the uterus has been demonstrated to be the luteolytic factor. In humans, luteal function may be influenced by local intraovarian eicosanoid production, which has been suggested to involve the paracrine interaction of local ovarian hormones such as oxytocin, noradrenaline, insulin and IGFs, to name but a few. Several lines of evidence have also implicated prostaglandins as an aetiological factor in ovarian pathological states such as seen in the OHSS. However, the bulk of clinical experimental evidence to date has failed to support this contention. Prostaglandin production has likewise been well characterized in the fallopian tube in both humans and animal species. Whereas a role for prostaglandins in tubal transport has been demonstrated with animal species such as the rabbit, several studies have failed to define a similar function in humans. More recently, direct injections of prostaglandin analogues into the fallopian tube and the corpus luteum have been shown to be efficacious as a treatment for ectopic pregnancy. Whether the primary mechanism of action involves effects on tubal musculature or corpus luteum function, or is simply a local vascular effect, remains to be demonstrated. Therefore, although the physiological role for eicosanoids in ovarian and tubal function remains unclear, particularly in the human, an increasing body of recent evidence has suggested an important paracrine function for this class of cellular mediators whose interaction with other more recently characterized local ovarian factors has only begun to be recognized.

Animals↗

The pharmacological activity of nitroxyl: a potent vasodilator with activity similar to nitric oxide and/or endothelium-derived relaxing factor.

Chemical oxidation of N-hydroxy-L-arginine (NOHA) and other N-hydroxyguanidines has been previously shown to generate either nitric oxide (NO) or nitroxyl (HNO), depending on the oxidative conditions. Because N-hydroxy-L-arginine has been demonstrated to be a biosynthetic intermediate in the oxidative conversion of arginine to endothelium-derived relaxing factor, the possible formation of HNO through a biological process was considered. This study, therefore, explores the biological activity of HNO as a possible effector molecule, and the results indicate that HNO is capable of eliciting vasorelaxation in both rabbit aorta and bovine intrapulmonary artery by a guanylate cyclase-dependent pathway. The pharmacological properties of HNO were very similar to those of endothelium-derived relaxing factor, and the possible relationship between HNO and endothelium-derived relaxing factor is discussed.

Animals↗

Acetylcholine vasodilation of resistance vessels in vivo may not entirely depend on newly synthesized nitric oxide.

Blood flow was measured in denervated hind limbs of pentobarbital-anesthetized cats. Intravenous NG-methyl-L-arginine, 3-30 mg/kg did not affect the magnitude or duration of vasodilation produced by intra-arterial acetylcholine. Intra-arterial NG-methyl-L-arginine induced hind limb vasoconstriction and reduced the duration but not the magnitude of the acetylcholine vasodilator response. The results suggest that acetylcholine-induced hind limb vasodilation may not entirely depend on newly synthesized nitric oxide.

Acetylcholine↗