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R Gross

Publications and source records attributed to R Gross.

At least 19 recordsLinked to original sources
PubMed1992

Adrenergic inhibition of insulin secretion involves pertussis toxin-sensitive and -insensitive mechanisms.

We studied the involvement of Bordetella pertussis toxin (PTX)-sensitive G proteins in the inhibition by adrenaline of insulin secretion from the isolated rat pancreas. The -90% inhibition induced by adrenaline (0.05 microM) was partially abolished after in vivo PTX pretreatment. The residual inhibitory effect of adrenaline in PTX-pretreated rats was suppressed by the alpha 2-adrenoceptor antagonist, yohimbine, but was not modified by the alpha 1-adrenoceptor antagonist, prazosin. Thus, the alpha 2-inhibitory effect of adrenaline on B-cells is mediated by both PTX-sensitive and PTX-insensitive mechanisms.

Analysis of Variance↗
PubMed1992

Binding characteristics of the new thromboxane A2/prostaglandin H2 receptor antagonist [3H]BAY U 3405 to washed human platelets and platelet membranes.

The new thromboxane A2 antagonist [3H]BAY U 3405 was characterized for its binding to washed human platelets and platelet membranes. In washed platelets the specific binding was reversible, selective and stereospecific, but not saturable. The dissociation constant (Kd) was 6 +/- 2.5 nM, the number of specific binding sites 1177 +/- 306 per platelet. Three structurally different thromboxane A2 (TXA2)/prostaglandin H2 (prostaglandin endoperoxide) (PGH2) receptor ligands completely inhibited the specific binding of [3H]BAY U 3405 in a concentration-dependent manner, indicating that the observed high affinity binding site is the TXA2/PGH2 receptor. In platelet membranes, however, specific [3H]BAY U 3405 binding showed saturability in addition to reversibility, selectivity, and stereospecifity. The Kd of the binding was 9.6 +/- 2.3 nM in kinetic studies and 8.7 +/- 3.7 nM in saturation studies, the inhibition constant (Ki) was 10 +/- 1.1 nM in displacement studies. The TXA2/PGH2 receptor agonists U 46619 and CTA2, and the antagonists Daltroban (BM 13505), I-PTA-OH and SQ 29548 all completely inhibited the specific binding of [3H]BAY U 3405 thus defining the observed binding site as the TXA2/PGH2 receptor. In conclusion, the data suggest that the previously reported TXA2 antagonism of BAY U 3405 is mediated by binding to a specific high affinity binding site of human platelets and platelet membranes that represents the TXA2/PGH2 receptor.

Blood Platelets↗
PubMed1992

pH dependency of the binding of [3H]BAY U 3405 and various non-labelled ligands to the thromboxane A2/prostaglandin H2 receptor of human platelet membranes.

[3H]BAY U 3405 was used to characterize the effect of acidic and alkaline pH values on the binding of the thromboxane A2/prostaglandin H2 (TXA2/PGH2) receptor of human platelet membranes. The specific binding of [3H]BAY U 3405 largely increased upon acidification up to pH 5.8. Saturation binding studies revealed an increase in binding affinity without change in the number of binding sites. At pH 7.4 the Kd was 8.7 +/- 3.7 nM (Bmax = 6.6 +/- 0.6 pmol/mg protein) compared to 1.2 +/- 0.2 nM (Bmax = 6.1 +/- 0.6 pmol/mg protein) at pH 5.8. A more than 10-fold higher rate of association was observed at pH 5.8 compared to pH 7.4, while the rate of dissociation showed only minor changes. The kinetically derived dissociation constant was 1 nM (pH 5.8) and 9.6 nM (pH 7.4). The pH dependency of the binding of structurally different non-labelled ligands to the TXA2/PGH2 receptor was evaluated by inhibition studies at pH 5.8 and pH 7.4. BAY U 3405, daltroban, CTA2, and U 46619 showed significantly higher affinities at pH 5.8. In contrast, I-PTA-OH and GR 32191 had a higher affinity at pH 7.4. No significant difference was seen with SQ 29548 at the observed pH values. A second protonable group within the molecules I-PTA-OH, GR 32191, and SQ 29548 might be responsible for the observed differences.

Blood Platelets↗
PubMed1992

Arachidonic acid induces an increase in the cytosolic calcium concentration in single pancreatic islet beta cells.

The insulin secretagogue D-glucose induces both accumulation of nonesterified arachidonic acid (35 microM) in pancreatic islets and a rise in beta cell cytosolic [Ca++]i. Arachidonate amplifies both voltage-dependent Ca++ entry in secretory cells and depolarization-induced insulin secretion. Here, arachidonate induced a biphasic rise in [Ca++]i of Fura-2AM loaded beta cells which increased with arachidonate concentration (5-30 microM), was reversed upon washout, and was unaffected by the arachidonate oxygenase inhibitor BW755C. The sustained phase of the rise was abolished by removal of extracellular Ca++ and amplified by depolarization with KCl. The accumulation of nonesterified arachidonate in islets stimulated by D-glucose may therefore promote the D-glucose-induced rise in beta cell [Ca++]i.

4,5-Dihydro-1-(3-(trifluoromethyl)phenyl)-1H-pyraz↗
PubMed1992

Evidence for a direct stimulatory effect of cibenzoline on insulin secretion in rats.

The effect of cibenzoline succinate, a new antiarrhythmic agent, was studied on insulin secretion in rats. Experiments were performed both in vivo and in vitro using two preparations: the isolated perfused pancreas and isolated islets. In anaesthetized rats, cibenzoline was able to increase plasma insulin levels and to reduce glycaemia. These effects were observed at 1 mg/kg i.v. in fed rats and at 3 mg/kg i.v. in fasted rats. In the isolated pancreas perfused in the presence of a slightly stimulating glucose concentration (8.3 mM), cibenzoline (2 and 6 microM) elicited a progressive and sustained insulin response in a concentration-dependent manner. In the presence of a non-stimulating glucose concentration (4.2 mM), cibenzoline was ineffective at 2 microM and slightly increased basal insulin release at 6 microM. In isolated islets incubated with 8.3 mM glucose, cibenzoline (6 and 20 microM) caused a concentration-dependent stimulation of insulin release. It is concluded that cibenzoline stimulates insulin secretion by a direct action on pancreatic B cells in rats.

Animals↗
PubMed1992

pH-dependent binding of the TXA2/PGH2-receptor of human platelet membranes to various ligands.

[3H]-BAY U 3405 was used to characterize the pH-dependency of the binding of various ligands to the TXA2/PGH2-receptor of human platelet membranes. Maximum binding of [3H]-BAY U 3405 is achieved at pH 5.8. In inhibition studies the ligands Daltroban, CTA2, and U 46619 also show a higher affinity at pH 5.8 compared to pH 7.4. In contrast, the ligands I-PTA-OH and GR 32191 have a higher affinity at pH 7.4. No difference is seen with SQ 29548. The ligands I-PTA-OH, GR 32191, and SQ 29548 have a second protonable group in common, which is thought to be the reason for the different pH-dependent binding.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗
PubMed1992

Functional analysis of the pertussis toxin promoter.

The expression of the pertussis toxin ptx operon is positively regulated in cis by a promoter region of about 170 base pairs and in trans by the bvg locus, which codes for the transcriptional activator protein BvgA. The promoter contains two direct repeats which are essential for its activity. When the position of these direct repeats relative to the transcription start point was changed, the activity of the promoter was strongly impaired. The repeated sequences therefore do not represent enhancer-like elements similar to those which have been identified in other positively regulated promoters; instead, the integrity of the whole promoter region seems to be an important feature of ptx regulation. A transcription interference assay was carried out to analyze in vivo binding of regulatory proteins to the ptx promoter. The results suggest that the direct repeats are the recognition sequence of a protein, which binds to them only under conditions in which the promoter is activated. In vitro DNA binding experiments with BvgA protein purified from an overproducing Escherichia coli strain were performed. However, no binding of BvgA to the ptx promoter was observed under conditions where binding of BvgA to the fha and bvg promoters occurred. This suggests that factors in addition to the bvg system are involved in the regulation of the Bordetella virulence regulon.

Bordetella pertussis↗
PubMed1992

Evidence for a direct inhibitory effect of PYY on insulin secretion in rats.

Peptide YY (PYY) has been shown to inhibit stimulated insulin secretion under in vivo conditions in the mouse, the rat, and the dog. In the present study, we investigated the effects of PYY on insulin secretion from the isolated perfused rat pancreas and isolated rat islets. In isolated pancreas perfused in presence of 8.3 mM glucose, PYY at 10(-10) and 10(-9) M, but not at 10(-8) M, inhibited insulin secretion. In the presence of 5.5 mM glucose, PYY (10(-9) M) did not modify basal insulin release but reduced the biphasic insulin response to arginine (10 mM). PYY also markedly reduced the pancreatic vascular flow rate; this effect was observed at all three concentrations tested in a dose-dependent manner. In isolated islets, glucose (15 mM)-stimulated insulin secretion was inhibited by PYY at 10(-7) M. We conclude that in the perfused rat pancreas, PYY inhibits insulin secretion and induces vasoconstriction without a causal relationship. In addition, our results on isolated islets suggest that the inhibitory action of PYY on insulin secretion is exerted through a direct islet action.

Animals↗
PubMed1992

Expression of bacterial cytotoxin genes in mammalian target cells.

We have studied the expression of the gene fragments encoding the enzymatically active portion of three bacterial cytotoxins: exotoxin A (ETA) of Pseudomonas aeruginosa, and pertussis toxin (PT) and adenylate cyclase toxin (CYA) of Bordetella pertussis, in sensitive mammalian target cells. Expression of active ETA and CYA was lethal to the producing cells and stable transfectants of Cos-1 cells containing the corresponding genes could not be obtained. The expression of the PTS1 subunit was tolerated by the producing mammalian cells. Since PT is cytotoxic because of ADP-ribosylation of G-proteins, we assume that the endogenously expressed PTS1 may not find the cellular target G proteins or PTS1 alone may not be sufficient for ADP-ribosylation of these proteins in vivo.

ADP Ribose Transferases↗
PubMed1992

Evidence for a glutamate receptor of the AMPA subtype which mediates insulin release from rat perfused pancreas.

1. The effect of L-glutamate has been studied on insulin secretion by the isolated perfused pancreas of the rat. The glutamate receptor subtype involved has been characterized. 2. In the presence of a slightly stimulating glucose concentration (8.3 mM), L-glutamate (5 x 10(-5)-4 x 10(-3) M) induced an immediate, transient and concentration-dependent insulin response. On the other hand, in the presence of a non stimulating glucose concentration (2.8 mM), L-glutamate (10(-3) M) did not modify the basal insulin secretion. 3. The three non-NMDA receptor agonists, kainate (10(-4)-10(-3) M), alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA, 5 x 10(-5)-10(-4) M) and quisqualate (5 x 10(-6)-5 x 10(-5) M) all provoked a transient and concentration-dependent insulin response from pancreas perfused with 8.3 mM glucose. Compared with glutamate, kainate exhibited a similar efficacy, whereas AMPA and quisqualate elicited only a 3 fold lower maximal insulin response. In contrast, NMDA (10(-4)-10(-3) M) was ineffective. 4. An antagonist of non-NMDA receptors, 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX; 5 x 10(-5) M) totally prevented the stimulatory effect of L-glutamate (4 x 10(-4) M) and kainate (2 x 10(-4) M). In contrast, the NMDA receptor antagonist, (+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclohepten-5,10-imine ((+) MK801) was without effect. 5. The insulin secretory effect of glutamate (4 x 10(-4) M) was not affected by atropine (3 x 10(-7) M) or tetrodotoxin (3 x 10(-6) M). 6. Quisqualate at a high maximally effective concentration (4 x 10(-4) M) inhibited glutamate (10(-3) M) or kainate (4 x 10(-4) M)-induced insulin release. 7. This study shows that L-glutamate stimulates insulin secretion in rat pancreas, by acting on an excitatory amino acid receptor of the AMPA subtype.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1992

P2y purinoceptor responses of beta cells and vascular bed are preserved in diabetic rat pancreas.

1. To investigate the effect of experimental diabetes on the P2y purinoceptor responses of pancreatic beta-cells and vascular bed, we used adenosine-5'-O-(2-thiodiphosphate) (ADP beta S), a potent and stable P2y agonist. This work was performed in the isolated perfused pancreas of the rat. 2. Diabetes was induced by streptozotocin (66 mg kg-1, i.p.). Five weeks after the induction of diabetes, on the day of pancreas isolation, the animals displayed marked hyperglycaemia (37.6 +/- 2.7 mM). Age-matched rats were used as controls. 3. Insulin response to a glucose stimulation from 5 to 10 mM was completely lost and stimulation of insulin release by the sulphonylurea, tolbutamide (185 microM), was drastically impaired in the diabetic pancreas (maximum responses were 1.5 +/- 0.4 and 7.0 +/- 1.4 ng min-1 for diabetic and age-matched rats respectively). 4. In contrast, in the diabetic pancreas ADP beta S (15 microM), infused in the presence of glucose 5 mM, elicited an immediate and significant insulin release similar to that observed in the age-matched pancreas (maximum responses were 7.6 +/- 1.5 and 6.7 +/- 1.3 ng min-1 respectively). This ADP beta S stimulating effect occurred independently of the glucose concentration (5, 8.3 and 28 mM) in the diabetic pancreas. On pancreatic vascular resistance, ADP beta S induced a similar vasodilatation in diabetic and age-matched rats. 5. In conclusion, ADP beta S retains its insulin stimulatory and vasodilator effects in experimental diabetes; P2y purinoceptors could therefore be considered as a new target for the development of antidiabetic drugs.

Animals↗
PubMed1992

Organ- and age-specific replication of polyomavirus in mice.

A novel organ- and age-specific pattern of polyomavirus DNA replication in mice is described. Two broadly defined classes of response to polyomavirus infection were observed: class I organs (mammary gland, bone, and skin) responded with high levels of replication in neonate mice and moderate levels in adults; class II organs (kidney, liver, and lung) responded with high levels in neonates and very low levels in adults. Thus, aging affected replication in all organs, and organ specificity was superimposed on this age-related decrease. We argue that the organ- and age-specific pattern likely reflects in part the activities of a multiplicity of general or tissue-specific, age-dependent transcription factors, which modulate viral replication or viral transcription or both. Interestingly, the majority of tumors in mice infected as neonates or as immunoincompetent adults originate in class I organs, suggesting that the ability to replicate in adult tissues is an important factor controlling polyomavirus oncogenesis. From the analysis of the infection process in adult mammary glands, a novel mode of polyomavirus infection emerged which contrasts with that derived from observations of tissue culture systems. A nonproductive infection was seen, characterized by very low levels of live virus (in the range of 10(-4) PFU per cell) and maintenance of the viral genome in an unintegrated, moderately replicating state. Maintenance of the viral genome was accomplished without integration into host cell DNA in all three tumor-prone organs, both prior to as well as beyond oncogenesis.

Aging↗
PubMed1992

The output and outcome of two types of formal health structures--health post and creche--for nutritional interventions for preschool children in two urban, low-income communities of Belo Horizonte, Brazil (1986).

In order to observe the nutritional and health status of pre-school children, the output and outcome of two formal health services -health post and creche- for this vulnerable group in two urban slum areas of Belo Horizonte, Brazil were studied in 1986. A total of 420 children were surveyed, 254 children randomly selected from the communities and 156 from three creches. Growth monitoring was not undertaken systematically, and mothers did not have growth control charts. When a child had diarrhea, mothers preferred to apply home remedies or to buy proprietary drugs rather than to consult medical personnel. 72% of mothers reported using ORT, and 11% suspending feeding completely. After three month of life, 50% of infants were receiving some breast milk. 75% of children were immunized completely. The prevalence of health and nutrition indicators such as anemia (50% vs. 30%), parasitosis (87% vs. 70%), and stunting (28% vs. 20%) was worse in the creches than in the communities.

Anemia↗
PubMed1992

The group work of the workshop.

The project aims at determining how aging influences nutritional status (as determined by measurements and indicators from clinical evaluation, anthropometric, laboratory, and biophysical tests) in a Chronic degenerative disease burden; 5) Use of drugs and medications (both Western and traditional) and 6) physical activity as the direct mediators of nutritional status of the elderly. A chain of causal determinants of each of the six mediators was developed. The primary target population is composed of persons from 60 to 75 years of age who are migrants to the metropolis from the same rural province. They share the same religious practices and ethic heritage in common and are free living (non-institutionalized) in the community. We shall enroll, in a stratified randomized manner (stratified for danger, urban community, and existence of participating second generation relative), 400 elderly persons in two groups: Group 1A: 100 males and 100 females who migrated more than 40 years earlier from the province to the metropolis; Group 1B: 100 males and 100 females who migrated less than 20 years earlier from the province to the metropolis. In a "controlled cross-sectional survey" format these represent two exposure groups: long-residency and short residency. A secondary contrast group will be gender specific, second generation relatives (sons or nephews for older men; daughters or nieces for older women), 400 in all, ie, one for each first generation (elderly) subject. They will range from 25 to 42 years of age and will also be living in low-income peri-urban communities of the metropolis. The tests from the overall battery of measurement indicators, appropriate to young adults, will be applied. The primary contrast is G1A vs G1B, aggregated or deseggregated by gender, using 1) relative risk based on long vs short urban exposure and 2) conventional parametric and non-parametric intergroup comparisons. The secondary contrast is within individual generation pairs (father-son, uncle-nephew; mother-daughter, aunt-nice) using conventional parametric or non-parametric tests for dependent variables.

Adult↗
PubMed1991

Diabetes alters the responses of glucagon secreting cells and vascular bed to isoprenaline and forskolin in vitro in rat pancreas.

Diabetes is known to disturb pancreatic glucagon secreting alpha cell function and blood flow control. In a previous study we could show that streptozotocin-induced diabetes suppressed adenosine stimulating effect on glucagon secretion and reduced its vasodilatory properties; since the nucleotide exerts these effects by activation of A2 purinergic receptors known to be positively coupled to adenylate cyclase, we investigated the effect of streptozotocin diabetes on the responses of alpha cells and vascular bed to stimulation of adenylate cyclase through 2 different ways: 1) with isoprenaline by activation of beta adrenergic receptor positively coupled to the enzyme; 2) with forskolin by direct activation of the catalytic unit of adenylate cyclase. In the isolated perfused pancreas of normal rats, isoprenaline (0.01 microM) or forskolin (1 microM) induced a +200 to +300% increase of glucagon secretion and a 20 to 30% increase of pancreatic vascular flow rate. In pancreas of 5-week diabetic rats, alpha cell responses to isoprenaline and forskolin were completely suppressed and the vasodilatory effects of both drugs were significantly reduced (-35 to -50%). Long term in vivo insulin treatment with glycaemia normalization was able to correct both defects. We can conclude that streptozotocin diabetes suppresses glucagon secretion and reduces pancreatic vasodilatation not only in response to activation of receptors positively coupled to adenylate cyclase but also to a direct activation of this enzyme.

Adenylyl Cyclases↗