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Biomedical subjects

E Gerlach

Publications and source records attributed to E Gerlach.

At least 37 records · Page 2Linked to original sources
PubMed1995

Nitric oxide accounts for postischemic cardioprotection resulting from angiotensin-converting enzyme inhibition: indirect evidence for a radical scavenger effect in isolated guinea pig heart.

The cardioprotective effect of angiotensin-converting enzyme (ACE) inhibitors in cardiac ischemia/reperfusion damage is assumed to result largely from inhibition of the enzymatic breakdown of endogenous bradykinin (BK). We assessed the role of nitric oxide (NO) in mediating the beneficial actions of BK and the possible mechanism of the effect of NO. We experimentally infringed myocardial function in a working guinea pig heart preparation by ischemia (15 min) and reperfusion. The parameter external heart work (EHW), determined before and after ischemia, served as criterion for quantitation of recovery. We assessed oxidative stress during reperfusion by measuring glutathione release in coronary venous effluent; lactate release was used as a measure of ischemic challenge. The principal ability of NO to scavenge oxygen radicals was separately investigated in a chemiluminescence (CL) assay with the NO-donor sodium nitroprusside (SNP) and lucigenin. The ACE inhibitor ramiprilat (RT 25 microM) improved postischemic function significantly (55% recovery of EHW vs. 29% for controls). BK 1 nM was even more cardioprotective (71% recovery). The NO-synthase inhibitor Ng-nitro-L-arginine (NOLAG 10 microM) inhibited the effects of RT and BK (18% recovery each). SNP (0.3 microM) improved recovery to 57%, the prostacyclin analogue iloprost (ILO, 0.1 and 3 nM) had no beneficial effect (21 and 20% recovery, respectively). With 8-bromo-cyclicGMP, a membrane-permeable cGMP analogue, function was not better than control (30% recovery). Release of glutathione during reperfusion was decreased by the three compounds known to increase NO concentration in the heart; lactate release was the same in all groups.(ABSTRACT TRUNCATED AT 250 WORDS)

Acridines↗
PubMed1995

No association of antibodies to glutamic acid decarboxylase and diabetic complications in patients with IDDM.

OBJECTIVE: To evaluate the association of antibodies to glutamic acid decarboxylase (GAD-ab) and diabetic complications (neuropathy, retinopathy, and nephropathy) in patients with insulin-dependent diabetes mellitus (IDDM). RESEARCH DESIGN AND METHODS: We examined the prevalence of GAD-ab (immunoprecipitation assay) and islet cell antibodies (ICAs) (indirect immunofluorescence) in a representative sample of IDDM patients (n = 146) with different disease duration (2-52 years, median 13.2 years). Of all patients characterized for the existence of diabetic complications, 56 of 146 had peripheral neuropathy, 24 of 142 had autonomic neuropathy, 67 of 141 had retinopathy, and 39 of 146 had nephropathy. RESULTS: GAD-ab (> 2 SD) were detected more frequently than ICA (> 5 Juvenile Diabetes Foundation units) in IDDM patients of different disease duration (GAD-ab+ 37% [54 of 146] vs. ICA+ 22% [32 of 146], P = 0.011; diabetes duration less than median: GAD-ab+ 47% vs. ICA+ 23%, P = 0.0046; diabetes duration greater than median: GAD-ab+ 27% vs. ICA+ 22%, P > 0.05). For GAD-ab and for ICA, respectively, no difference was observed in frequency of positivity or titers between patients with or without diabetic complications. CONCLUSIONS: Both GAD-ab and, to a lesser extent, ICA persist for a long time in several individuals. This persistence is not related to diabetic neuropathy or any other diabetic complication.

Adolescent↗
PubMed1995

Preischaemic as well as postischaemic application of a calcium antagonist affords cardioprotection in the isolated guinea pig heart.

OBJECTIVE: The aim was to answer the following questions: (1) Does treatment with calcium antagonists have to be begun before ischaemia or is postischaemic application also protective? (2) When applied before ischaemia, do calcium antagonists have to depress preischaemic cardiac function in order to elicit protection? (3) Is cardioprotection a matter of improved reflow or do the agents influence the degree of oxidative injury during reperfusion? METHODS: Isolated working guinea pig hearts underwent ischaemia (15 min) and reperfusion (15 min). The calcium antagonist gallopamil was given either before (0.1 nM and 1 nM) or after ischaemia (0.1 nM) during early reperfusion (first 5 min). Recovery was defined as postischaemic compared to preischaemic external heart work, expressed in percent. Oxidative stress was assessed by the release of glutathione (GSH). Lactate release served as a measure of the ischaemic challenge. The ability of gallopamil to scavenge oxygen radicals directly was investigated in an in vitro chemiluminescence assay. RESULTS: Pump function of control hearts recovered to only 28% after reperfusion. Pretreatment with 0.1 and 1 nM gallopamil improved recovery to the same extent (48.7% and 43.4%, respectively); however, postischaemic application of 0.1 nM gallopamil afforded equal protection (45.4% recovery). Only the higher concentration of 1 nM gallopamil depressed preischaemic external heart work (by 11%). During earliest reperfusion (1-5 min), release of GSH only tended to be lower in treated hearts. During the subsequent minutes of reperfusion (5-15 min), release of GSH was significantly less in hearts postischaemically treated with 0.1 nM gallopamil (40 pmol.min-1 v 940 pmol.min-1 for controls). In contrast, ischaemia-induced lactate release did not differ between the groups. Gallopamil did not scavenge reactive oxygen species in vitro. CONCLUSIONS: Short term postischaemic application of the calcium antagonist gallopamil is almost as effective at restoring pump function as preischaemic application which, in turn, does not have to depress preischaemic cardiac function in order to elicit protection. A reduction of oxidative stress during reperfusion seems to contribute to the beneficial effects of postischaemic application of gallopamil, but a direct oxygen radical scavenging activity of gallopamil is not involved.

Animals↗
PubMed1994

Stimulation of endothelial adenosine A1 receptors enhances adhesion of neutrophils in the intact guinea pig coronary system.

OBJECTIVE: The primary aim was to determine the action of pathophysiologically relevant adenosine concentrations (0.1-1 microM) on adhesion of neutrophils to coronary endothelium. Further aims were to evaluate the nature and localisation of the adenosine receptor involved, and to assess the effect of endogenous adenosine. METHODS: Adhesion was studied in isolated perfused guinea pig hearts by determining the number of cells emerging in the coronary effluent after intracoronary bolus injections of 600,000 neutrophils prepared from guinea pig or human blood. The system was characterised by the use of the proadhesive stimulus thrombin. RESULTS: A 5 min infusion of adenosine (0.1-0.3 microM) or the A1 receptor agonist N6-cyclopentyladenosine (CPA, 0.01 microM) significantly increased adhesion from about 20% (control) to 30%. This effect was prevented by the A1 receptor antagonist dipropyl-8-cyclopentylxanthine (DPCPX, 0.1 microM). It was not diminished by cessation of adenosine infusion 90 s prior to neutrophil injection. At a higher concentration of adenosine (1 microM), adhesion did not seem to be enhanced. However, coinfusion of the A2 receptor antagonist 3,7-dimethyl-1-propargylxanthine (DMPX, 0.1 microM) with 1 microM adenosine unmasked the A1 action, adhesion rising to 39%. Adenosine had a quantitatively identical effect on adhesion of human neutrophils. Total ischaemia of 15 min duration raised adhesion of subsequently applied neutrophils to 35%. This effect was completely blocked by DPCPX, as well as by ischaemic preconditioning (3 x 3 min). Preconditioning raised initial postischaemic coronary effluent adenosine from about 0.8 microM to 1.5 microM. CONCLUSIONS: The findings suggest a bimodal participation of adenosine in the development of postischaemic dysfunction by an endothelium dependent modulation of neutrophil adhesion. Stimulation occurs via endothelial A1 receptors at submicromolar adenosine levels, whereas cardioprotection by adenosine may in part relate to the use of pharmacologically high concentrations of adenosine or enhanced endogenous production after preconditioning.

Adenosine↗
PubMed1994

Bradykinin accounts for improved postischemic function and decreased glutathione release of guinea pig heart treated with the angiotensin-converting enzyme inhibitor ramiprilat.

We investigated the role of bradykinin (BK) in cardioprotection elicited by angiotensin-converting enzyme (ACE) inhibition is isolated guinea pig heart performing pressure-volume work. Cardiac output (CO), coronary blood flow (CBF), and external heart work (EHW) were determined before and after ischemia and reperfusion (15 min each). Furthermore, the glutathione (GSH) content of hearts and the release of glutathione in coronary venous effluent were measured, as was lactate production. Addition of the ACE-inhibitor ramiprilat (RT) to the perfusate throughout the experiment improved postischemic function significantly (55% recovery of EHW for 25 microM RT vs. 30% for controls). RT was cardioprotective even if only given at onset of reperfusion (50% recovery). BK (0.1 and 1 nM) was similarly beneficial (55 and 76% recovery of EHW, respectively). The BK2-receptor antagonist HOE 140 (10 nM) inhibited the RT effect and attenuated the effect of 1 nM BK. Total CBF during reperfusion, lactate production, intracellular levels of GSH, and release of oxidized GSH (GSSG) did not differ among the groups. In contrast, release of reduced GSH during the first 5 min of reperfusion was considerably influenced by pharmacologic intervention, correlating inversely with postischemic heart function. Coapplication of Hoe 140 prevented the changes in GSH release. Our results demonstrate that BK, formed endogenously in the heart, is responsible for cardioprotection by the ACE inhibitor RT in isolated guinea pig heart and decreases GSH release during reperfusion. The exact mechanisms leading to hemodynamic improvement and metabolic changes by BK remain unclear.

Adenosine Diphosphate↗
PubMed1994

Coexpression of the human TNF receptors TR60 and TR80 in insect cells: analysis of receptor complex formation.

For investigation of a possible physical interaction between the two human tumor necrosis factor receptors, TR60 (type I) and TR80 (type II), the baculovirus expression system was used. Each of the receptors was expressed as a membrane-integrated protein in insect cells, able to specifically bind the two ligands, tumor necrosis factor (TNF) and lymphotoxin (LT alpha). Typically, about 150,000 membrane receptors per cell could be detected 40 h after infection, exerting high affinity ligand binding capacity with Kd values virtually identical to that of human cell lines. The baculovirus system allowed coexpression of both TNF membrane receptors at very high and about equal numbers to investigate the existence of heteromultimeric receptor complexes, either formed spontaneously or ligand induced. Neither saturation binding studies nor immunoprecipitation experiments gave an indication for the existence of TNF receptor heteromers. These data are in accordance with the current view of TNF signaling, in which homonultimerization, rather than heteromer formation of TNF receptors is the initial activating event.

Animals↗
PubMed1994

Expression of complete human IFN-gamma receptor and its extracellular domain in insect cells: purification and characterization of the recombinant proteins.

We here describe an efficient procedure for overexpression and purification of recombinant complete human interferon-gamma (IFN-gamma) receptor (IFN-gamma-R) and its extracellular fragment employing a baculovirus (BV) expression system. Infection of Sf 158 cells with recombinant BV results in membrane expression of high affinity IFN-gamma-R (Kd 1.6 x 10(-10) M), with approximately 10(6) molecules/cell 40 h postinfection. Solubilized, affinity-purified IFN-gamma-R and a secreted extracellular domain of IFN-gamma-R were compared for ligand-binding capacity and antagonistic activity in an IFN-gamma bioassay. Our results show that the complete receptor has a 2.5-fold higher ligand affinity and a 15-fold higher IFN-gamma in vitro-neutralizing capacity in an in vitro virus protection assay as compared to the extracellular fragment. This suggests that the transmembrane and cytoplasmic domains of IFN-gamma-R contribute to stability and/or enhance formation of biologically active receptor complexes in solution.

Animals↗
PubMed1993

Adenosine contributes to neutrophil-mediated loss of myocardial function in post-ischemic guinea-pig hearts.

Tissue injury associated with myocardial ischemia is assumed to largely result from the toxic effects of active oxygen species generated by accumulated polymorphonuclear leukocytes (PMNs). Recent reports have indicated that adenosine can interfere with the PMN function in vitro. The potential of adenosine to influence PMN-mediated myocardial tissue injury was assessed using a model of ischemia-reperfusion injury developed in the isolated working guinea-pig heart perfused with homologous PMNs. After an initial work phase, hearts were subjected to 30 min low-flow ischemia (1 ml/min) in the absence and presence of PMNs. Work was resumed after 15 min reperfusion in a non-working mode (Langendorff). Adenosine in the coronary effluent reached a maximum of 0.2 microM during low-flow ischemia. Recoveries of external heart work and cardiac output were reduced from about 80% to about 40% by PMNs. Infusion of adenosine deaminase (ADA, 5 U/ml), theophylline (50 microM) or the selective A1-antagonist dipropyl-8-cyclopentylxanthine (0.1 microM) prevented this effect. Furthermore, application of adenosine (0.1 microM) in combination with PMNs also resulted in a loss of pump function, even in the absence of a direct ischemic stimulus. The data indicate that adenosine contributes to post-ischemic, PMN-mediated damage in the isolated working guinea-pig heart model by a receptor-mediated action.

Adenosine↗
PubMed1993

Relative efficacy of tazobactam, sulbactam and clavulanic acid in enhancing the potency of ampicillin against clinical isolates of Enterobacteriaceae.

Three beta-lactamase inhibitors were combined with ampicillin in a fixed 2:1 ratio. The activity of ampicillin was enhanced by tazobactam and by clavulanic acid, and to a lesser extent by sulbactam when tested against fresh clinical isolates of Enterobacteriaceae. At a concentration of 8 micrograms/ml, ampicillin alone inhibited 49.6% of 2,434 consecutive isolates of enteric bacilli compared to 81% inhibited by ampicillin combined with tazobactam or clavulanic acid and 69.3% inhibited by the sulbactam/ampicillin combination. A four-fold or greater reduction in ampicillin MICs was observed in comparable numbers of isolates with all three combinations, but the most marked effects were seen with strains that were highly resistant to ampicillin.

Ampicillin↗
PubMed1993

Ampicillin-sulbactam susceptibility testing criteria.

In vitro studies in five different medical centers documented the susceptibility of 2,440 consecutive isolates of the Enterobacteriaceae against ampicillin-sulbactam disks of different potencies. For determination of MICs, both 2:1 or 1:1 ratios were used as long as the concentrations of sulbactam at the breakpoints remained the same, i.e. MIC < or = 16/8.0 micrograms/ml or < or = 8.0/8.0 micrograms/ml for the susceptible category. Disks containing 10 micrograms of ampicillin and 10 micrograms of sulbactam are still to be preferred with interpretive criteria of > or = 15 mm for susceptible and < or = 11 mm for resistant (MIC > or = 64/32 micrograms/ml or > or = 32/32 micrograms/ml). The reliability of the disk test actually diminished when the amount of sulbactam in the disk was increased.

Ampicillin↗
PubMed1993

[Interaction of blood and the vascular wall: hemostatic aspects].

The coagulation enzyme thrombin, a serine protease like all other coagulation factors, plays a central role in the hemostatic processes engaged after injurious events. It induces, with particular efficacy, the aggregation of blood platelets (primary hemostasis) and accounts, via splitting of fibrinogen to fibrin, for the event actually responsible for the coagulation of blood (secondary hemostasis). As is well-known, thrombin itself is generated by a cascade of activation events involving various coagulation factors (F). In this respect the "tissue factor" (TF, formerly known as thromboplastin), in combination with F VIIa, attains decisive significance, not only in the extrinsic pathway of coagulation (activation of F X-->Xa), but also in the intrinsic pathway (activation of F IX-->IXa). Under physiological circumstances, platelet aggregation and coagulation are restricted to the area of the vascular lesion, since the surrounding intact endothelium inhibits an intraluminal spreading of both processes. These "antithrombotic" features of the endothelium encompass antiaggregatory mechanisms (formation and release of prostacyclin [PGI2], adenosine, EDRF [NO], degradation of ADP and other nucleotides mediated by ecto-nucleotidases) as well as anti-coagulatory properties (formation and release of "tissue factor pathway inhibitor" [TFPI], which blocks the coagulation cascade by joining F Xa, TF and F VIIa into an inactive complex, thrombomodulin--thrombin induced activation of protein C, which, together with protein S, inactivates F Va and F VIIIa, thereby attenuating further generation of thrombin, and the heparan sulfate-enhanced activation of antithrombin III and heparin-cofactor II).(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Different endothelial mechanisms involved in coronary responses to known vasodilators.

The role of the endothelium in mediating flow responses to acetylcholine (ACh), bradykinin (BK), adenosine (Ado), and the poorly hydrolyzable ATP-derivative beta,gamma-methylene-ATP (MeATP) was evaluated in the intact coronary system of the isolated perfused guinea pig heart. In the presence of superoxide anions, known to inactivate the endothelium-derived relaxing factor nitric oxide (NO), only steady-state dilatation induced by ACh (1 microM) was fully inhibited and that of BK (0.1 nM) attenuated. Similar effects were obtained with methylene blue and N omega-nitro-L-arginine; however, the latter also reduced the actions of Ado (0.1 microM) and MeATP (0.5 microM). Conversely, perfusion with the NO precursor L-arginine (10 microM) resulted in a potentiated relaxation by ACh, whereas steady-state responses to BK and Ado remained unchanged. Pretreatment of hearts with hydroxyl radicals (.OH) elevated vascular permeability. Under this condition, flow increases induced by ACh, BK, and Ado were enhanced by 130, 89, and 47%, respectively, whereas the effect of MeATP (0.5 microM) was reduced by 45%. Preexposure of hearts to the oxidant hypochlorous acid (HOCl) prevented dilatations by ACh, BK, MeATP and Ado (0.1 microM), and the response to Ado (5 microM) was reduced by 68%; postischemic hyperemia was attenuated. Glyburide, an inhibitor of ATP-sensitive K+ channels, halved the flow response to infused Ado (0.1 and 5 microM), inhibited MeATP, and abolished reactive hyperemia. We conclude that in the guinea pig coronary system ACh, BK, MeATP, and Ado (0.1 microM) induce endothelium-dependent vasodilatation, some step(s) of the signal transmission being vulnerable to oxidative attack by HOCl.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

No evidence for the existence of preformed antibodies against hydroxyethyl starch in man.

Using the highly sensitive ELISA technique for detecting anti-hydroxyethyl starch (HES) antibodies in man sera from 1,056 patients were analyzed. Patients of both sex, who had never had any prior contact with HES, were included in the study. In none of the cases could any titer of HES-reactive antibodies be detected. These data suggest that in man preformed HES-reactive antibodies do not exist or are extremely rare. In any case, unlike dextran-reactive antibodies, they should not have a high clinical importance. The mechanism behind the very rarely observed anaphylactoid reactions after HES application is still unknown.

Adult↗
PubMed1991

[Are the radical scavenging properties of ACE inhibitors with sulfhydryl groups in therapeutically effective concentrations of quantitative significance?].

The chemiluminescence reaction elicited from luminol in the presence of hydroxyl radicals was concentration-dependently suppressed by captopril, indicating efficacious radical scavenging. As to be expected, ACE inhibitors lacking free sulfhydryl groups (ramipril, enalapril) were inactive. However, the endogenous scavenger and anti-oxidant uric acid proved to be far superior to captopril, when concentrations of both were compared that are realized in vivo. A substantial augmentation of endogenous scavenging ability during therapy with captopril thus seems unlikely. In a model of standardized myocardial hypoxia (isolated working heart of the guinea pig with 30 min low flow perfusion) captopril, ramiprilat and uric acid equally improved post-hypoxic heart function. There was no cardioprotective action of captopril in excess of that accountable for by inhibition of ACE. It seems possible that ACE (kininase II) inhibitors exert cardioprotection via elevated tissue levels of kinins: bradykinin also improved heart performance after low flow perfusion and bradykinin-induced coronary dilatation was markedly enhanced in the presence of ramiprilate, reflecting attenuated degradation by endothelial kininase II.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1989

Uric acid as radical scavenger and antioxidant in the heart.

Uric acid (UA) is released from the heart of many species, including man, and its site of formation has been shown to be the microvascular endothelium. Since UA reacts with oxygen radicals in vitro, experiments were conducted on guinea pig hearts perfused with Krebs-Henseleit buffer (KHB) to evaluate whether the formation of UA could afford protection from damage by radicals and oxidants. The following results were obtained: (1) Upon addition of the hydroxyl radical scavenger DMSO to the perfusate, the coronary rate of release of endogenous uric acid was increased relative to the precursor purines. (2) UA was degraded during passage through the coronary system and also in KHB in vitro after addition of substances generating hydroxyl radicals or hypochlorite. Superoxide (O2-) radicals did not seem to react directly with UA, though UA concentration-dependently quenched the chemiluminescence generated from luminol in the presence of O2- and OH radicals. (3) Coronary dilation by acetylcholine (Ach) and sub-microM concentrations of adenosine, induced by both via endothelial mechanisms, was attenuated after prolonged inhibition of endothelial UA formation by allopurinol. Furthermore, the effect of Ach but not of adenosine proved acutely sensitive to methylene blue and O2-, substances known to inactivate EDRF. This finding suggests involvement of EDRF in Ach-mediated, but not in adenosine-induced dilatation of the intact coronary system. Exogenously applied UA prevented the impairment of vascular responses to Ach and adenosine caused by allopurinol, and to Ach upon generation of O2-. (4) Hearts performed more pressure-volume work and exhibited greater functional stability when perfused with KHB supplemented with UA in a physiological concentration.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholine↗
PubMed1989

Effects of chronic treatment with adrenaline or propranolol on platelet function and c-AMP levels in the rat.

Most of our knowledge about the modulation of platelet function by catecholamines is based on observations of acute in vitro actions. Little is known about the effects of chronically elevated or reduced adrenergic stimulation of the platelets. We therefore treated rats for 8 weeks with either adrenaline or the beta-blocker propranolol. Adrenaline (0.5 mg.kg-1.d-1) continuously administered from subcutaneously implanted osmotic mini pumps caused an increase in the sensitivity of the platelets towards ADP as stimulating agent. In contrast, chronic application of propranolol (10 mg.kg-1.d-1) via the drinking water led to a reduction in platelet aggregability. For animals treated with adrenaline, in accordance with the results of the aggregation experiments, the levels of c-AMP found in platelet rich plasma were reduced, both basally (by 33%) and after stimulation of platelet adenylate cyclase with prostaglandin E1 (by 39%). For the propranolol treated animals, the basal c-AMP concentrations remained unchanged. The levels of c-AMP attained after stimulation with prostaglandin E1 were diminished to a similar extent as for the adrenaline treated animals (by 38%). Although the in vitro addition of adrenaline to platelet rich plasma causes a beta-adrenoceptor mediated inhibition of platelet aggregation in the rat, the simulation seen after chronic adrenaline exposure in vivo, which is associated with decreases in both basal and stimulated c-AMP levels, suggests a functional preponderance of alpha-adrenoceptors over beta-adrenoceptors on the rat platelets. Although intraplatelet metabolic changes (blockade of stimulated c-AMP formation) after chronic application of propranolol should have resulted in enhancement of platelet aggregability, an inhibition of aggregation was observed.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Diphosphate↗