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H Weiner

Publications and source records attributed to H Weiner.

At least 127 records · Page 7Linked to original sources
PubMed1989

Liver mitochondrial aldehyde dehydrogenase: in vitro expression, in vitro import, and effect of alcohols on import.

An in vitro expression plasmid (pGRAP) that contained the cDNA coding for the rat mitochondrial aldehyde dehydrogenase precursor was constructed, mRNA was synthesized then translated, and the in vitro synthesized precursor of aldehyde dehydrogenase was used in an in vitro import assay. As expected the 19 amino acid signal peptide of the precursor allowed import of the precursor into rat liver mitochondria. This in vitro system was used to examine the effect of alcohols on import. It was found that the alcohols (ethyl, butyl, hexyl, and octyl) tested inhibited the import of the aldehyde dehydrogenase precursor. Pretreatment of the mitochondria with alcohol was responsible for the inhibition. The inhibition appeared to be relatively specific for pre-aldehyde dehydrogenase as the precursor of ornithine transcarbamylase was still imported in the presence of alcohols. Of potential physiological significance was finding that ethanol inhibited import in a dose-response fashion; 50% inhibition occurred at 75 mM, a concentration achievable during the ingestion of alcohol. In addition, the concentrations of alcohols required to produce an inhibitory effect on import decreased as the hydrocarbon chain length of alcohols increased. The inhibitory effect of alcohols appeared to be specific as other solvents examined did not inhibit import. We postulate that alcohols may perturb the mitochondrial membrane and affect the receptor-translocator necessary for the import of the aldehyde dehydrogenase precursor.

Aldehyde Dehydrogenase↗
PubMed1989

Primary structures of rat and bovine liver mitochondrial aldehyde dehydrogenases deduced from cDNA sequences.

The cDNA coding for rat liver mitochondrial aldehyde dehydrogenase was cloned and sequenced. It contained an open reading frame of 1557 bp. Of the deduced 519 amino acid residues, 19 were proposed to correspond to the signal peptide necessary to allow the protein to enter the mitochondria [Farrés, J., Guan, K.-L. and Weiner, H. (1987) Biochem. Biophys. Res. Commun. 150, 1083 - 1087]. The sequence of the 500 amino acid residues comprising the mature subunit was 96% identical to that of the corresponding human liver mitochondrial enzyme. The longest cDNA isolated coding for the bovine liver enzyme contained sequence information corresponding to residues 72 - 500 of the rat or human enzyme. The deduced protein sequence of these residues was 94% identical to those of the human enzyme. Over 60% of the bases in the coding region of the rat cDNA were G and C. These residues were clustered non-randomly and two potentially stable stem-loop structures could be calculated as existing in the mRNA. One would be found in the region coding for amino acid residues 64 - 79; a similar secondary structure could be responsible for the existing truncated cDNAs from the bovine-cDNA libraries. Comparisons with the amino acid sequences of other aldehyde dehydrogenases support the suggestion that the NAD-binding domain may be located in the middle portion of the mature enzyme.

Aldehyde Dehydrogenase↗
PubMed1989

Natural killer cell activity in vasopressin-deficient rats (Brattleboro strain).

Several lines of evidence suggest that the neuropeptide vasopressin is involved in the regulation of the immune system. We explored this possibility by comparing the cytotoxic activity of natural killer (NK) cells in Brattleboro (DI) rats, which are homozygous for diabetes insipidus and lack vasopressin, and Long-Evans (LE) rats, the strain from which DI rats were derived. Additionally, we compared the effects of swim stress, morphine administration and vasopressin replacement on NK cell activity in these two strains. In DI rats, NK cell activity, determined by a standard 4-h chromium-release assay, was significantly higher than in LE rats. Both swim stress and morphine administration suppressed NK activity in DI and LE rats. There was no difference in the level of suppression between the two strains. Vasopressin replacement normalized water intake in DI rats, but had no significant effect on NK cell activity. DI rats exhibited lower plasma corticosterone levels, which were not elevated by vasopressin replacement. The results suggest that the lack of vasopressin in DI rats elevates baseline NK cell activity, probably via mechanisms that are secondary to the vasopressin deficiency (e.g. lower corticosterone levels). Neither vasopressin nor other hormones affected by vasopressin deficiency seem to be involved in the acute modulating effects of stress and morphine on NK cells.

Animals↗
PubMed1989

Presence of cytosolic aldehyde dehydrogenase isozymes in adult and fetal rat liver.

A colony of Wistar-strain rats bred at Purdue University was composed of animals with two different isozyme patterns of liver cytosolic aldehyde dehydrogenase (EC 1.2.1.3, ALDH) as determined by isoelectric focusing. One cytosolic isozyme pattern had a major activity band with a pI = 5.8 and a minor activity band at pI = 6.2. The other pattern contained three major isozymes with pI values of 5.3, 5.4 and 5.6 along with the pI 6.2 isozyme and a trace of the 5.8 one. The 5.8 and 6.2 isozymes were recognized by antibodies produced against horse and beef liver cytosolic ALDH, whereas the set of three (5.3-5.6) were not. The cytosolic isozymes were inhibited by low levels of disulfiram and had Km values for acetaldehyde in the 100 microM range, properties typical for cytosolic ALDHs. All animals contained the same isozymes of liver mitochondrial ALDH. These were a major activity with a pI = 5.2 and minor activities associated with isozymes of pI = 6.4 and 6.6. These isozymes were recognized by antibodies produced against pure horse and beef liver mitochondrial ALDHs. Both cytosolic and mitochondrial ALDHs were found in fetal liver as early as day 15 of gestation. The total activity for mitochondrial ALDH increased between day 15 and day 21 whereas that for cytosolic ALDHs remained relatively constant during development. It appeared that both cytosolic and mitochondrial ALDH were present by at least the third trimester and could afford the fetus some protection against the toxic action of endogenous or exogenous aldehydes.

Acetaldehyde↗
PubMed1989

Potentiation of cigarette craving and satisfaction by two types of meals.

This study investigated the effects of consuming different meal types on cigarette craving and satisfaction. Twelve heavy smokers were tested on three evenings. Each day, they smoked three cigarettes and received either a solid meal, an equicaloric liquid meal, or no meal. Subjects smoked the first cigarette one half hour after the experiment began, the second immediately after finishing the meal, and the third 35 min after smoking the second cigarette. Cigarette craving increased most after the solid meal, less so after the liquid meal, and least after no meal. Subjects reported that the cigarette after the solid meal tasted the best, was the most satisfying, enjoyable, and desirable as compared to the cigarette following no meal at all. These results indicate that there is a robust postprandial enhancement of cigarette smoking after a solid meal but less after a liquid meal.

Adult↗
PubMed1989

Social confrontation "stress" in aggressive fish is associated with an endogenous opioid-mediated suppression of proliferative response to mitogens and nonspecific cytotoxicity.

We have previously shown that social confrontation between aggressive fish (e.g., Tilapia) produces a suppression of several immunological parameters--nonspecific cytotoxicity and mitogen-stimulated proliferation in pronephric leukocytes--in the subordinate fish. By using the opioid antagonist, naltrexone, we now demonstrate indirectly that this immunosuppression is in part mediated by the endogenous opioid system. Evidence is presented that naltrexone-mediated reversal of immunosuppression may be limited to the populations of the cytotoxic and T-cell lineages. The proliferation response to lipopolysaccharide is unaffected by naltrexone. Our data also demonstrate that serum from subordinate (immunosuppressed) fish is immunosuppressive in normal fish--an effect that can be reversed by naltrexone. These results support a link between the neuroendocrine and immune systems in fish, the lowest vertebrate.

Animals↗
PubMed1989

Predictable and unpredictable shock stimulates gastric contractility and causes mucosal injury in rats.

The effects of tailshock on gastric contractility and lesions were investigated in rats exposed to 100 1-mA tailshocks while confined inside plastic tubes. A light preceded each shock in one group and was randomly presented with respect to shock in the other. Following the session, animals were given 3 hr of rest before being sacrificed. Contractility of the corpus of the stomach was measured by means of chronically implanted extraluminal force transducers. Contractility was measured in 10-min blocks and analyzed by computer. Lesions were quantified by inspection; quantitative histology was performed on corpus and antrum sections. Signaled (n = 13) and unsignaled (n = 17) shock stimulated high-amplitude gastric contractions in fasted rats, which continued for 2 hr after the shock session. Cumulative contractile activity (1.5-hr shock plus 2-hr rest) in shocked animals was twice that in restrained and unrestrained control animals (n = 19, p less than .05), and contractile activity had a 30%-40% greater average amplitude than after a meal. Compared with unrestrained controls, shocked rats had visibly more mucosal injury (2.2 +/- 0.5 mm2 vs. 0.1 +/- 0 mm2). Larger cumulative contractile activity was associated with a larger area of erosions (r = .36, p less than .05). Frequency and duration of contractions did not distinguish between shocked and unshocked groups. We conclude that in rats, signaled and unsignaled tailshock stimulates persistent, high-amplitude gastric contractions and is associated with injury of the mucosa of the stomach.

Animals↗
PubMed1989

Medullary sites for TRH analogue stimulation of gastric contractility in the rat.

Medullary sites at which the stable thyrotropin-releasing hormone (TRH) analogue, RX 77368 (p-Glu-His-[3,3'-dimethyl]-Pro-NH2), stimulates gastric contractility were investigated in rats under urethan anesthesia. The peptide analogue was microinjected in 50-100 nl of volume unilaterally into various brain stem nuclei using glass micropipettes (50 microns). Gastric contractility was recorded continuously with acutely implanted strain-gauge force transducers and traces analyzed by computer. RX 77368 (2.6-77 pmol) microinjected into the dorsal vagal complex (DVC) dose dependently stimulated gastric contractility. Peptide action (77 pmol) had a rapid onset with a peak response at 15 min, continued for 45 min after the microinjection, and was blocked by vagotomy. TRH microinjected into the DVC had a shorter duration of action. RX 77368 (0.7-77 pmol) microinjected into the nucleus ambiguus also dose dependently stimulated gastric contractions. In contrast, microinjection of RX 77368 (77 pmol) into the hypoglossal nucleus had no effect. These findings demonstrate that the DVC and nucleus ambiguus are sites of action for TRH-induced stimulation of gastric contractility in the rat. The effect is dose dependent, long lasting, site specific, and vagally mediated. These results are consistent with a possible physiological role for medullary TRH in the vagal regulation of gastric contractility.

Animals↗
PubMed1989

Serotonin depletion potentiates gastric secretory and motor responses to vagal but not peripheral gastric stimulants.

Vagal stimulation is known to release gastrointestinal serotonin. The effect of depletion of serotonin stores on vagally stimulated gastric acid secretion and motility was studied in rats. Pretreatment of rats with parachlorophenylalanine (p-CPA) did not alter basal gastric acid and serotonin secretion but produced a 57% reduction in the intraluminal gastric release of serotonin and a 43 to 100% potentiation of the gastric acid secretory response elicited by intracisternal injection of the stable thyrotropin-releasing hormone analog, RX 77368, in conscious pylorus-ligated rats or urethane-anesthetized rats with an acute gastric fistula. Dose-response studies revealed that the gastric acid secretion induced by submaximal but not high doses of RX 77368 was elevated significantly by p-CPA pretreatment. p-CPA also enhanced the gastric acid output produced by submaximal, but not high doses of the vagal stimulant baclofen, [beta-(p-chlorophenyl)-gamma-aminobutyric acid]. In contrast, p-CPA pretreatment had no effect on gastric acid secretion stimulated by bethanechol, histamine or pentagastrin. Selective depletion of central serotonin stores by pretreatment with the neurotoxin 5,7-dihydroxytryptamine given alone, or combined with parachloroamphetamine did not alter RX 77368-stimulated gastric acid secretion. In addition, gastric contractility stimulated by intracisternal injection of RX 77368 was significantly enhanced by p-CPA but not by 5,7-dihydroxytryptamine pretreatment, whereas the contractile response to carbachol was not altered by p-CPA pretreatment. These results suggest that depletion of peripheral but not central serotonergic stores potentiate gastric acid secretion and contractility induced by vagally, but not peripherally acting gastric stimulants. Thus, peripheral serotonin may exert an inhibitory tone on vagally stimulated gastric acid secretion and motility in the rat.

Animals↗
PubMed1989

Psychoendocrinology of anorexia nervosa.

An extensive literature exists on the pathophysiology of anorexia nervosa (AN). This article reviews succinctly the disturbances in the reproductive biology of AN; they exemplify the fragility of the physiologic mechanisms that underlie reproductive function. They pose the following question: Why is this crucial function so easily disrupted given that the species would not survive without it?

Amenorrhea↗
PubMed1988

Sequence of the signal peptide for rat liver mitochondrial aldehyde dehydrogenase.

The cDNA coding for the signal peptide of rat liver mitochondrial aldehyde dehydrogenase was sequenced. The deduced amino acid sequence of the signal peptide was MLRAALSTARRGPRLSRLL. From this sequence an amphiphilic helix which had a high hydrophobic moment could be constructed. A comparison to the published cDNA sequence of human mitochondrial aldehyde dehydrogenase revealed great sequence identity and allowed us to make some predictions regarding the primary structure of the human signal peptide.

Aldehyde Dehydrogenase↗
PubMed1988

Identification of the cysteine residue in the active site of horse liver mitochondrial aldehyde dehydrogenase.

Aldehyde dehydrogenase catalyzes the oxidation of aldehydes to acids through the formation of a covalent intermediate. It has been postulated that a cysteine residue could be acting as the active site nucleophilic group. Although N-ethylmaleimide was found to react with many cysteines it was possible by doing the reaction in the presence of chloral hydrate, a substrate analog which functions as a competitive inhibitor, to label cysteine at position 49 in the horse liver mitochondrial enzyme. The dehydrogenase activity was lost as the residue was modified, consistent with the possibility that the residue was an integral component of the active site of the enzyme. Cysteines at positions 162 and 369 also could be modified. It is suggested that cysteine 162 may function as part of a site capable of hydrolyzing nitrophenyl acetate. Details of the second site will appear in the accompanying paper (Tu, G. C., and Weiner, H. (1988) J. Biol. Chem. 263, 1218-1222). It appeared that the substrate-binding domain was in the N-terminal portion of the enzyme while the coenzyme binding domain was in the C-terminal portion. During this investigation 133 of the 500 residues of the horse liver enzyme were sequenced. These showed about 95% sequence identity with those of the human enzyme. Inasmuch as both beef and rat liver enzymes also share 95% identity with the human enzyme it can be expected that the results found with the horse liver enzyme can be applicable to all mammalian aldehyde dehydrogenase.

Aldehyde Dehydrogenase↗
PubMed1988

Evidence for two distinct active sites on aldehyde dehydrogenase.

Aldehyde dehydrogenase can catalyze the hydrolysis of esters such as p-nitrophenyl acetate as well as oxidize aldehydes to acids. It has not been proven unequivocally that the two reactions occur at the same active site. In the accompanying paper (Tu, G. C., and Weiner, H. (1988) J. Biol. Chem. 263, 1212-1217) evidence was presented which showed that cysteine at position 49 was at the active site for the dehydrogenase reaction. Evidence also was presented which showed that cysteine located at position 162 was susceptible to modification by N-ethylmaleimide. It was shown here that the two activities of the enzyme can be differently protected from inactivation by substrate analogs. Furthermore, aldehydes were found to be poor inhibitors against the esterase reaction while ester was a good inhibitor against the dehydrogenase reaction. In addition, it was possible to modify cysteine 49 with N-ethylmaleimide but not find inhibition of the esterase reactivity until cysteine 162 was modified. It appears that horse liver aldehyde dehydrogenase has two separate active sites per subunit. The data fit a model where ester can be hydrolyzed at both sites but that aldehyde oxidation occurred only at position 49.

Aldehyde Dehydrogenase↗
PubMed1988

Plasma cortisol and natural killer cell activity during bereavement.

Natural killer cell (NK) activity, which is important in the defense against tumors and viral infections, is reduced in women undergoing conjugal bereavement. The relationship between NK activity and plasma cortisol was investigated in three groups of subjects: women who were anticipating the death of their husbands, women whose husbands had recently died, and controls. Bereaved women showed reduced NK activity and increased plasma cortisol levels as compared to controls. Anticipatory bereaved women also showed significant reductions in NK activity, but had levels of plasma cortisol comparable to those of controls. The reduction of NK activity during anticipatory and actual bereavement cannot be explained solely on the basis of increased cortisol secretion.

Female↗
PubMed1988

Corticotropin-releasing factor acts centrally to suppress stimulated gastric contractility in the rat.

The effects of intracisternal (i.c.) and intravenous (i.v.) administration of corticotropin-releasing factor (CRF) on gastric contractility stimulated by i.c. injection of the TRH analog RX77368 [p-Glu-His-(3,3'-dimethyl)-Pro-NH2], 2-deoxy-D-glucose (2DG) and i.v. infusion of carbachol were evaluated in rats under urethane anesthesia. Gastric contractility was monitored using acutely implanted extraluminal force transducers sutured to the corpus of the stomach. I.c. injection of CRF (6.3-210 pmol) resulted in a dose dependent suppression of gastric contractility stimulated by RX77368 (260 pmol) and 2DG (6 mg). Gastric inhibitory response to i.c. CRF was rapid in onset and lasted at least 45 min. Carbachol (200 mg/kg/h)-induced stimulation of gastric contractility was not modified by i.c. injection of CRF. The stimulation of contractility caused by both i.v. carbachol and i.c. 2DG were completely inhibited by atropine (1 mg/kg, i.v.). CRF (210 pmol) given i.v. suppressed RX77368-stimulated gastric contractions, but was less than 1/10 as potent as administered i.c. I.v. CRF (210 pmol) did not alter 2DG- or carbachol-induced gastric contractions. These results demonstrate that the i.c. administration of CRF acts within the brain to inhibit gastric contractility elicited by vagus-dependent mechanisms.

Animals↗