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Biomedical subjects

A Sastre

Publications and source records attributed to A Sastre.

At least 55 records · Page 3Linked to original sources
PubMed1989

Isomeric yohimbine alkaloids block calcium-activated K+ channels in medullary thick ascending limb cells of rabbit kidney.

The alpha2-adrenergic antagonist yohimbine (YOH) and the closely related isomers corynanthine (COR) and rauwolscine (RAU) caused brief interruptions in current characteristic of a fast blocker Ca2+-activated K+ channels in cultured medullary thick ascending limb (MTAL) cells. The apparent dissociation constants (Kapp) for COR, YOH, and RAU, respectively, at the intracellular face of the channel in the presence of 200 mM K+ are 45 +/- 1, 98 +/- 2, and 310 +/- 33 microM. The Kapp for COR on the extracellular side also in the presence of 200 mM K+ was much greater at 1.6 +/- 0.17 mM. Increasing K+ on the same side as the blocker relieves the blocking reaction. The Kapp for the alkaloids varies with K+ in a manner quantitatively consistent with K+ and the alkaloids competing for a common binding site. Finally, blocking by the charged form of these alkaloids is voltage dependent with changes in Kapp of 86 +/- 7 and 94 +/- 6 microM per e-fold change in voltage for blockers applied either from the inside or outside. The alkaloids block at an electrical distance similar to tetraethylammonium, suggesting that the site within the channel pore of these molecules may be similar.

Animals↗
PubMed1989

Glutamate toxicity in a neuronal cell line involves inhibition of cystine transport leading to oxidative stress.

Glutamate binds to both excitatory neurotransmitter binding sites and a Cl(-)-dependent, quisqualate- and cystine-inhibited transport site on brain neurons. The neuroblastoma-primary retina hybrid cells (N18-RE-105) are susceptible to glutamate-induced cytotoxicity. The Cl(-)-dependent transport site to which glutamate and quisqualate (but not kainate or NMDA) bind has a higher affinity for cystine than for glutamate. Lowering cystine concentrations in the cell culture medium results in cytotoxicity similar to that induced by glutamate addition in its morphology, kinetics, and Ca2+ dependence. Glutamate-induced cytotoxicity is directly proportional to its ability to inhibit cystine uptake. Exposure to glutamate (or lowered cystine) causes a decrease in glutathione levels and an accumulation of intracellular peroxides. Like N18-RE-105 cells, primary rat hippocampal neurons (but not glia) in culture degenerate in medium with lowered cystine concentration. Thus, glutamate-induced cytotoxicity in N18-RE-105 cells is due to inhibition of cystine uptake, resulting in lowered glutathione levels leading to oxidative stress and cell death.

Animals↗
PubMed1989

Anaphylaxis produced by rubber glove contact. Case reports and immunological identification of the antigens involved.

A study was carried out on six subjects suffering from anaphylactic reactions including asthma, rhinoconjunctivitis, contact urticaria and hypotension following contact with rubber gloves. All of them showed skin sensitivity and serum specific IgE to various preparations of natural latex, and not to other chemicals used in glove manufacturing. Immunoblotting techniques demonstrated that at least four soluble polypeptides from natural latex had the ability to bind specific human IgE. The results suggested that natural latex proteins present in rubber gloves can cause severe specific hypersensitivity reactions.

Adolescent↗
PubMed1988

Glutamate cytotoxicity in a neuronal cell line is blocked by membrane depolarization.

To understand better the proximate mechanism involved in the excitotoxic response to L-glutamate (Glu), we have exploited the Glu receptor present in the N18-RE-105 neuroblastoma-embryonic retinal hybrid cell line. These cells undergo lysis dependent on extracellular Ca2+ when exposed to Glu. We now report that the depolarizing action of Glu is not responsible for its cytotoxic effects. Furthermore, depolarization of these cells with elevated K+, ouabain or veratridine does not cause cytotoxicity but rather protects against the cytotoxic effects of Glu. Our results may implicate a role for voltage-sensitive Ca2+ channels (VSCCs) in cytotoxicity, and depolarization-induced inactivation of VSCCs (Nature (Lond.), 316 (1985) 440-443) as a protection against Glu receptor agonists. Our findings demonstrate a clear dissociation between depolarization and the neuronal degeneration caused by Glu.

Animals↗
PubMed1988

Calcium-dependent glutamate cytotoxicity in a neuronal cell line.

Membranes from the neuroblastoma x embryonic retina cell hybrid cell line, N18-RE-105, bind L-[3H]glutamate with a pharmacologic profile consistent with a 'quisqualate-type' brain L-glutamate receptor. We describe here the cytotoxic effect of L-glutamate receptor agonists on intact N18-RE-105 cells. Cytotoxicity was quantitated by measurement of the release of the cytosolic enzyme, lactate dehydrogenase, into the culture medium after addition of L-glutamate and its analogs to the cell culture medium. L-Glutamate (10 mM) and its confirmationally restricted analogs, quisqualate (1 mM) and ibotenate (10 mM), caused cell lysis. In contrast, similar analogs which do not bind to N18-RE-105 cell membranes (kainic acid, N-methyl-D,L-aspartic acid and gamma-aminobutyric acid) were not cytotoxic. L-Glutamate-induced cytotoxicity was eliminated when calcium-free medium was used. Addition of inorganic or organic calcium channel antagonists also reduced the cytotoxicity of L-glutamate, even when 1.8 mM calcium was present in the medium. Cadmium chloride (10 microM) completely blocked L-glutamate toxicity, whereas manganese chloride (150 microM) and lanthanum chloride (25 microM) reduced toxicity by greater than 50%. Dihydropyridine voltage-sensitive calcium channel agonists or antagonists, had little or no significant effect on L-glutamate-induced toxicity. In contrast, the verapamil derivatives, D600 and D888, and the diltiazem derivative, MDL 12,330A reduced L-glutamate toxicity by greater than 50%. These results suggest that a subtype of voltage-sensitive calcium channels is involved in the mechanism of L-glutamate receptor mediated cytotoxicity in this cell line.

Calcium↗
PubMed1988

Contributions of alpha 1- and alpha 2-adrenoceptors to contractile response in canine blood vessels.

Strips of canine saphenous vein, inferior vena cava, and femoral artery were studied isometrically in vitro to compare quantitatively the alpha 1- and postsynaptic alpha 2-adrenoceptor contributions to the contractile force generated by l-norepinephrine (NE). Effects mediated by each receptor type were measured independently by quantitative blockade of virtually all alpha 1-receptors with prazosin, or alpha 2-receptors with rauwolscine. Appropriate concentrations of the antagonists were calculated from dissociation constants previously determined by binding or competition with [3H]prazosin or [3H]rauwolscine in tissue homogenates. The contribution of alpha 1-adrenoceptors was larger than that of alpha 2-receptors in all vessels. The alpha 2-type was responsible for 38% of the maximum unblocked response to NE in saphenous vein, 32% in vena cava, and 28% in femoral artery. The occupation-response relationship for alpha 1-receptors was almost linear, without the marked upward convexity reported in some other vessels. alpha 2-Occupation-response curves were convex towards the occupation axis, with a relatively small response at low levels of occupation.

Animals↗
PubMed1988

Cholinergic receptors and contraction of smooth muscle in canine portal vein.

The properties of cholinergic receptors in homogenates of canine portal vein were determined with the radioligand [3H]quinuclinidyl benzilate [( 3H]QNB), and correlated with the functional responses of that vessel to carbamylcholine (CCh) in vitro. [3H]QNB bound to a single population of sites in the homogenates, with a dissociation constant (Kd) of 125 pM (+/- 19), and a total receptor capacity of 24.4 fmol/mg of protein, which corresponded to 224 (+/- 67.3) fmol/g wet weight of the initial vascular wall sample. Competition of CCh with [3H]QNB in eight experiments revealed two binding sites of different affinity, present in about equal numbers with Kd = 0.48 microM and 31 microM (in the absence of 5'-guanylylimidodiphosphate), and Kd = 1.5 microM and 42 microM (in the presence of 100 microM 5'-guanylylimidodiphosphate). Vascular strips in vitro contracted in response to CCh, with a threshold of approximately 0.3 microM, ED50 of 1.87 microM and maximum response of 344 g/cm2 (65% of the maximum response to l-norepinephrine). Strips precontracted by l-norepinephrine or by high potassium concentrations were slightly (4%) relaxed by 0.1 microM CCh, but further contracted by higher concentrations. The contractile responses were not altered by removal of the endothelium, and were blocked by atropine but not hexamethonium. The results demonstrate the existence in this vein of muscarinic cholinergic receptors that mediate smooth muscle contraction, and the receptor properties resemble those reported in cardiac muscle and brain.

Algorithms↗
PubMed1987

Intranasally and orally administered antihistamine treatment of experimental rhinovirus colds.

Although antihistamines are widely used for symptomatic treatment of seasonal (allergic) rhinitis, the role of histamines in the pathogenesis of infectious rhinitis is not clear. Two antihistamines, orally administered chlorpheniramine maleate (CM) and diphenhydramine hydrochloride (DH) administered by intranasal spray, were used under double-blinded, randomized, placebo-controlled conditions to assess both tolerance and efficacy in volunteers with experimental rhinovirus (RV) colds. In the initial trial, CM (4.0 mg per treatment) was ingested 4 times daily for 4 days beginning 24 h after intranasal inoculation of RV type 29 in susceptible volunteers. In the second trial, DH was sprayed intranasally 4 times daily (2 mg per treatment) for 5 days beginning 24 h after inoculation of RV type 39. Clinical colds occurred in 60% of the CM group (n = 13) and 73% of the placebo (n = 15) in the first study, and in 66% of the DH group (n = 12) compared with 81% in the placebo group (n = 11) in the second. Both CM and DH were well tolerated but had no significant effects on nasal symptoms or nasal mucus production. The findings provide additional evidence against an important role for histamine in the pathogenesis of nasal symptomatology in rhinovirus colds.

Administration, Intranasal↗
PubMed1986

Serum angiotensin converting enzyme levels in patients with alpha 1-antitrypsin variants.

Serum angiotensin converting enzyme levels were measured in 184 subjects having either MM, MZ, ZZ, or MS Pi-types of alpha 1-antitrypsin. Elevated angiotensin converting enzyme levels were detected in 31 percent of the patients with the MZ Pi-type, 20 percent of the patients with the ZZ Pi-type, and 20 percent of the patients with the MS Pi-type compared with 1.33 percent of those with normal MM Pi-type. The mean serum angiotensin converting enzyme levels were also significantly higher in those with the MZ, ZZ, and MS Pi-types. Multiple family members of two families were found to have both the Z variant and angiotensin converting enzyme elevations, suggesting the possibility of a genetic linkage. Alpha 1-antitrypsin deficiency must be added to the list of disease states potentially associated with elevated serum angiotensin converting enzyme levels.

Adult↗
PubMed1986

Alpha 1-antitrypsin Pi-types in 965 COPD patients.

To study further the role of intermediate alpha 1-antitrypsin (AAT) deficiency in chronic obstructive pulmonary disease (COPD), AAT Pi-types and serum-trypsin-inhibitory-capacity (STIC) were measured in 965 patients with COPD. Heterozygosity of the Z variant was the major cause of intermediate AAT deficiency (primarily the MZ phenotype), accounting for 8.0 percent of the patients compared to 2.9 percent of control subjects (p less than .0005). ZZ homozygosity was detected in 1.9 percent of the patients, compared to 0.04 percent of control studies performed by others (none was present in our own control group of 1,380 subjects). The mean age for MS or MZ patients did not differ from that of the COPD patients as a whole, whereas the ZZ homozygotes were younger (55.9 +/- 9.8 vs 65.3 +/- 7.5 years). These results resemble those of a previous study in 66 male veterans with pulmonary emphysema suggesting that the MZ phenotype, or intermediate AAT deficiency in general, probably does predispose to the development of COPD. However, the prevalence of AAT deficiency in COPD patients is small (approximately 10 percent). The number with an MS phenotype was not increased in this group of COPD patients.

Adult↗
PubMed1986

An angiotensin-converting enzyme (ACE) inhibitor in human serum. Increased sensitivity of the serum ACE assay for detecting active sarcoidosis.

Measurement of serum angiotensin-converting enzyme (ACE) is extremely useful as an aid in the diagnosis and longitudinal evaluation of patients with sarcoidosis. We have detected a human serum ACE-inhibitor which affects the ACE level obtained by activity measurements. The effect of the inhibitor can be eliminated by a mere eight-fold dilution of the serum sample with physiologic saline. We recommend that serum ACE be performed with 1:8 dilutions of serum to eliminate the effect of the ACE inhibitor. The inhibitor has a MW above 50,000 daltons, and reversibility of inhibition by dilution appears to be ion dependent. Dialysis of an inhibitor-containing serum sample against saline causes the inhibition to become irreversible, allowing the distinction between alinearity of the assay vs an inhibitor effect when a serum has ACE activity greater than 50 units/ml. The source of the serum ACE-inhibitor remains to be determined.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1986

Serum angiotensin-converting-enzyme in rabbits with and without pulmonary granulomatosis. Granulomatosis induced with complete-Freund's-adjuvant or BCG.

An animal model with pulmonary, non-caseating, granulomatosis was successfully reproduced in rabbits utilizing intravenous injections of complete-Freund's-adjuvant (CFA) or BCG. Granuloma formation from CFA injections was limited to the lungs, but BCG injections caused systemic granulomatosis (lung, liver and spleen) irrespective of whether the BCG was dissolved in saline or in incomplete-Freund's-adjuvant (IFA). Serum angiotensin-converting-enzyme (ACE) levels began to rise approximately 15 days following the intravenous injection of CFA, but similar rises following the injection of IFA without granuloma formation suggest that the ACE rise was not necessarily reflective of granuloma formation in rabbits. BCG in saline caused no rise in serum ACE, but when suspended in IFA the ACE rise was comparable to that resulting from IFA alone. However, the ACE level of broncho-alveolar-lavage cells obtained from rabbits 21 days post-injection of CFA was 7-fold higher than that of lavaged cells obtained 42 days post injection, correlating with an acute alveolitis occurring early during granuloma formation. These studies confirm the usefulness of intravenous injections of CFA or BCG for inducing a granulomatous state, but show that the assay of serum ACE is not useful for reflecting the granulomatous state in rabbits. In addition, serum ACE levels are 2-5 X higher in rabbits than in humans and tend to fluctuate spontaneously following delivery of the animals to the laboratory.

Animals↗
PubMed1985

Standardization of the prick test. A comparative study of three methods.

The different needles and methods used in the prick test give rise to disparate results. This has significance when carrying out multi-centre studies and when using the technique in the standardization of allergenic extracts. With test reliability as our objective, prick tests were carried out on 30 subjects: 10 patients sensitive to Dermatophagoides pteronyssinus received a glycerine extract of known allergenic potency, 10 healthy individuals 2.5% codeine phosphate in a glycerine solution, and another 10, histamine 1/1000. The total prick tests per individual was 27 with each of the needles employed (Allergy Pricker, the Morrow-Brown needle and Insulin needle in accordance with Pepy's procedure). The tests were carried out systematically by three different testers, and the total number of prick tests performed was 810. With the Allergy Pricker, no differences were observed among results obtained by the same tester, nor when the results of the three testers were compared. With the Morrow-Brown needle, the results varied in the same person and from one tester to another, and on many occasions the test was negative. With the Pepys method, no falsely negative results were obtained, but there was considerable variation in the size of the wheal. The variation coefficient is 41% with the Allergy Pricker, and 115% and 64% with the Morrow-Brown and Pepys method, respectively. In conclusion, the results obtained clearly indicate that the highest degree of reproducibility is obtained with the Allergy Pricker.

Adolescent↗
PubMed1985

Correction of amino-acid imbalance in postoperative portosystemic shunts.

We report on 22 cirrhotic patients suffering from portal hypertension and bleeding esophageal varices. Sixteen of them underwent H mesocaval shunt with internal jugular vein graft and 6 spleno-renal shunts. Mortality was 13.2%. All the patients were treated postoperatively with parenteral and oral nutritional support with solutions containing no aromatic amino-acids, following Fischer's formula. We determined Fischer's index (formula; see text); normal value 2.67 +/- 0.22) on the day of operation (1.6 +/- 0.22), on the 8th postoperative day (2.32 +/- 0.49), and on the 15th postoperative day (1.85 +/- 0.37). Only 1 patient suffered from hepatic encephalopathy in the immediate postoperative period. Three months after operation Fischer's index was 1.20 +/- 0.11 and only 2 patients with an index below 1 suffered from chronic encephalopathy. We conclude that there is a correlation between Fischer's index and the presence of post-shunt hepatic encephalopathy. The proposed policy of parenteral nutrition improves the values of Fischer's index in the immediate postoperative period and we believe that nutritional support should be included routinely as an adjuvant therapy in portosystemic shunts in cirrhotic patients.

Adult↗