Biomedical subjects

B Penke

Publications and source records attributed to B Penke.

At least 73 records · Page 4Linked to original sources

PubMed1994

Conformational mapping of amyloid peptides from the putative neurotoxic 25-35 region.

The secondary structure of amyloid beta A(25-35) and its deletion analogues was studied by circular dichroism (CD), Fourier transform infrared (FTIR) spectroscopy and molecular dynamics calculation. Data of our comparative CD and FTIR measurements in trifluoroethanol suggest that beta A(25-35)NH2 has a preferred beta-sheet conformation. Contrary to this beta A(31-35)NH2 tends to adopt a beta-turn conformation. Based on the comparable neurotoxic effect of beta A(25-35)NH2 and beta A(31-35)NH2 the neurotoxicity likely involves the same 31-35 core sequence and the "biologically active conformation" is a beta-turn rather than a beta-sheet structure.

Amino Acid Sequence↗

PubMed1994

Lateral mobility of the antagonist-occupied V2 vasopressin receptor in membranes of renal epithelial cells.

The lateral mobility of membrane integral receptors has been implicated as playing a significant role in signal transduction. The adenylate cyclase-coupled vasopressin V2 receptor has been shown to be highly laterally mobile in membranes of LLC-PK1 renal epithelial cells at physiological temperature using a fluorescent vasopressin agonist, with lateral mobility of the V2 receptor proposed to play a role in both adenylate cyclase activation and ligand induced receptor internalization and down-regulation. This study reports the synthesis and characterization of two new fluorescent antagonists [(beta-mercapto-beta,beta-cyclopentamethylene propionic acid)1,D-Tyr2,Ile4,Lys9(N6-fluoresceinylaminothiocarbonyl )]AVP (FL-AVP-anta) and [(beta-mercapto-beta,beta-cyclopentamethylene propionic acid)1,D-Tyr2,Ile4,Lys9(N6-tetramethylrhodamylaminothioca rbonyl)]AVP (TR-AVP-anta) for the V2 receptor. The latter was used to determine the parameters of lateral mobility of the V2 receptor in the non-activated antagonist-occupied form. Using fluorescence photobleaching techniques, results were largely comparable to those for agonist-occupied receptor, indicating high mobility at 37 degrees C. Antagonistic properties of the V2 receptor ligands are apparently not related to decreased receptor lateral mobility. Photobleaching measurements, however, did show that in contrast to V2 agonist, V2 antagonist did not induce receptor immobilization due to aggregation with time at 37 degrees C, indicating that this could be of mechanistic importance in the internalization process.

Amino Acid Sequence↗

PubMed1994

T cell recognition of the posttranslationally cleaved intersubunit region of influenza virus hemagglutinin.

The influenza virus hemagglutinin is synthesized as a single polypeptide chain, but upon maturation it will posttranslationally be modified by a host cell related trypsin-like enzyme. The enzymatic cleavage attacks the so-called intersubunit region of the molecule giving rise to covalently linked HA1 and HA2 subunits. An I-Ed-restricted T cell epitope was identified in the highly conserved intact intersubunit region of the influenza virus hemagglutinin. T cell recognition of a 25-mer synthetic peptide comprising the intact intersubunit region does not require further processing and the elimination of the intervening Arg residue coupling the fusion peptide to the C-terminal segment of HA1 does not abolish the T cell activating capacity. The fine specificity pattern of a T cell hybridoma similar to that of the polyclonal T cell response demonstrates that a single T cell receptor is able to recognize peptides of different sizes representing not only the uncleaved but also the cleaved form of this hemagglutinin region. Based on specificity studies the epitope was localized to the C-terminal 11 amino acids of the HA1 subunit. The cross-reactivity of peptide-primed T cells with influenza virus infected antigen-presenting cells shows that fragments comprising the identified epitope of the intersubunit region can be generated as a result of natural processing of the hemagglutinin molecule. As antigen-presenting cells are lacking the enzyme which is responsible for the posttranslational modification of newly synthesized hemagglutinin molecules, the role of immature viral proteins in immune recognition is discussed.

Amino Acid Sequence↗

PubMed1994

The lymphocyte transformation test with sulphadiminidum and with prealbumin-sulphadiminidum complex in patients with drug allergy.

Earlier results (the absence or a decreased level of prealbumin [PA] in the serum, a higher proportion of PA-bearing lymphocytes as antigen-binding cells, and the presence of PA in the eluate of lymphocytes in patients with drug allergy) permitted the conclusion that, besides its other relevant functions, PA may also behave as a hapten carrier protein. The lymphocyte transformation test (LTT) involving measurement of 3H-thymidine uptake was therefore carried out with sulphadiminidum (SA) alone, with the PA-SA complex, and with the albumin-SA complex in patients with proven (3 cases) or suspected (63 cases) sensitivity to SA. In all 3 cases with proven drug allergy to SA and in 5 of the 63 suspect cases, the LTT was positive in response to both the drug alone and to the PA-SA complex (The albumin-SA complex gave positivity in only one case). In 5 of the 63 suspect cases, the LTT was positive only in response to the PA-SA complex. It is suggested that the sensitivity of the LTT could be increased if the test could be run with a complex of a protein (hapten carrier?) and a drug.

Drug Eruptions↗

PubMed1994

The intersubunit region of the influenza virus haemagglutinin is recognized by antibodies during infection.

The influenza virus haemagglutinin has an important role in the infectious cycle of the virus and carries multiple B and T cell epitopes. It is synthesized as a single polypeptide chain but viral infectivity depends on its post-translational enzymatic cleavage. The cleavage site of a trypsin-like enzyme responsible for this modification is found in the most conserved intersubunit region of the molecule. In this study the role of this region in antibody recognition was investigated. Synthetic peptides comprising the intact and cleaved forms of the intersubunit segment were used to examine the specificity of virus- or peptide-induced antibodies. The immune response elicited by viral infection resulted in the appearance of antibodies capable of neutralizing the virus without interfering with its binding to the receptor. A monoclonal antibody (MoAb) of such functional properties was shown to recognize the intact intersubunit region both in the uncleaved haemagglutinin molecule and in a 25-mer synthetic peptide comprising the intact intersubunit region. Specificity and functional studies revealed the conformation-dependent recognition of the C-terminal segment of the haemagglutinin 1 subunit by this MoAb. The binding of the antibody was shown to inhibit the trypsin-mediated cleavage of the haemagglutinin molecule and the membrane fusion event. The enzymatic cleavage of the haemagglutinin was demonstrated to abolish antibody recognition of the infective virus suggesting an escape mechanism mediated by the functional destruction of this highly conserved region. The synthetic peptide corresponding to the intact intersubunit region is characterized by an ordered structure and is able to elicit an antibody response in BALB/c mice while its subfragments are nonimmunogenic. Furthermore, this peptide elicited a protective immune response demonstrated by in vivo experiments.

Amino Acid Sequence↗

PubMed1994

A new synthesis of Rink's polymer, 4-(2',4'-dimethoxyphenylhydroxymethyl)phenoxymethylpolystyrene .

For solid-phase peptide synthesis, 2,4-dimethoxy-4'-hydroxbenzhydrol linker was prepared via lithium borohydride reduction of 2,4-dimethoxy-4'-hydroxybenozophenone. The potassium salt of the linker was coupled to chloromethylpolystyrene. This method proved to be better than use of the cesium salt. This new synthesis gave a polymer with appropriate structure and a good degree of substitution.

Amino Acids↗

PubMed1994

Affinity chromatographic method for the extraction of ocytocin from human and rat plasma.

A procedure based on affinity chromatography for the extraction of ocytocin is described. Ocytocin antibody, covalently attached to a magnetizable iron suspension, retains its ability to bind ocytocin from blood plasma. The conjugate can be reused several times. Recovery of ocytocin was 60-70%, and the detection limit for the extraction assay was about 1 pg per tube. The specificity of the extraction method was characterized by HPLC/RIA on specimens extracted from blood plasma. The extracted material was found to be chomatographically identical with the reference ocytocin preparation. Comparison of RIA data for the same samples extracted by immunoextraction (y) and adsorption (x) showed a linear regression of y = 0.83 x + 2.12, r = 0.81, n = 17.

Animals↗

PubMed1994

Specific radioimmunoassay of oxytocin in rat plasma.

Sensitive, specific and reproducible radioimmunoassay (RIA) was developed for the measurement of oxytocin (OXT) in rat blood plasma after various extraction methods. The assay is based on an antiserum raised against OXT in rabbit. The sensitivity, affinity constant, and cross-reactivity of the antiserum were determined. The 125I-labelled OXT for RIA was produced by chloramine-T method and purified with high pressure liquid chromatography (HPLC). Two extraction procedures were employed: 1. adsorption to an artificial silicate, Lichroprep Si 60 (Merck); 2. immunoextraction of the hormone applying a magnetic bearer covered with purified antibodies against OXT. The specificity of the extraction methods was characterized in comparative HPLC/RIA studies of specimens extracted from blood plasma in different ways. The basal level of the peptide measured after the extraction with thermally activated Lichroprep Si 60 or after the immunoextraction method was found to be 9.6 +/- 2.3 pg/ml (mean +/- S.E.) and 15.3 +/- 0.9 pg/ml (mean +/- S.E.), respectively. Various well known factors (ether exposure, hyperosmotic stress and suckling) appeared to be potent peripheral stimuli of OXT release, and thus indicated the suitability of the RIA method for the measurement of OXT in blood plasma.

Adsorption↗

PubMed1993

Immunohistochemical visualization of a metabotropic glutamate receptor.

The immunocytochemical localization of the recently cloned metabotropic glutamate receptor 1 alpha (mGluR1 alpha) was demonstrated with a C-terminus specific antibody in rat cerebellar cortex. This antibody detects a 138-140 kDa major, and a 46 kDa minor band in membrane preparations of rat cortex and cerebellum. mGluR1 alpha immunoreactivity (mGRi) was present in Purkinje and basket cells. Purkinje cell dendritic spines and their postsynaptic membranes showed selective labelling. Presynaptic membranes, parallel fibres and glial processes were devoid of mGRi. It is suggested that the selective postsynaptic localization of this receptor at the dendritic spines of Purkinje cells serves as the morphological basis for long term depression processes in the molecular layer of the cerebellar cortex.

Amino Acid Sequence↗

PubMed1993

Role of endogenous corticotropin-releasing factor in mediation of neuroendocrine and behavioral responses to cholecystokinin octapeptide sulfate ester in rats.

The possible involvement of endogenous corticotropin-releasing factor (CRF) in the anxiogenic and pituitary-adrenal-axis-activating effects of cholecystokinin octapeptide sulfate ester (CCK 8) was investigated in rats. Intracerebroventricularly (i.c.v.) administered CCK 8 induced an anxiogenic response in an elevated plus-maze test, and enhanced the plasma corticosterone level. Pretreatment with different dilutions (1:10, 1:20 and 1:100, i.c.v.) of CRF antiserum and different doses of a CRF receptor antagonist, alpha-helical CRF (ahCRF, 0.001-1.0 microgram, i.c.v.) prevented the anxiogenic response to CCK 8 in a dose-dependent manner. None of the doses of CRF antiserum or ahCRF alone produced any alteration in either the elevated plus-maze paradigm or corticosterone level in saline-treated control rats. The results strongly suggest that the anxiogenic and hypothalamo-pituitary-adrenal-activating effects of CCK 8 are mediated via CRF.

Animals↗

PubMed1992

The cocaine-induced elevation of plasma corticosterone is mediated by endogenous corticotropin-releasing factor (CRF) in rats.

The role of endogenous corticotropin-releasing factor (CRF) in the cocaine-induced corticosterone response was investigated by using the immunoneutralization and receptor blockade of endogenous CRF. Pretreatment with different dilutions (1:5, 1:10 and 1:20, i.c.v.) of CRF antibody and different doses of an antagonist for CRF receptors, alpha-helical CRF9-41 (alpha h-CRF, 0.001-1.0 micrograms, i.c.v.), dose-dependently prevented the cocaine-induced increase in corticosterone level. These results support the hypothesis that the activation of the hypothalamo-pituitary-adrenal (HPA) axis by cocaine is mediated through the release of endogenous CRF.

Animals↗

PubMed1992

Conformational and functional properties of peptides covering the intersubunit region of influenza virus hemagglutinin.

The functionally active part of influenza virus hemagglutinin was investigated through the synthesis of a series of peptides representing different parts of the intersubunit region. Secondary structure prediction, circular dichroism and Fourier transform infrared spectroscopic studies were undertaken to investigate the secondary structure of these peptides. The peptide fragments were found to adopt multiple conformations, depending on their concentration in solution, the presence of the non-ionic detergent octyl-beta-D-glucoside and the polarity of the solvent. The results of biological studies with these peptide fragments are discussed in relation to their conformation, as inferred from the spectroscopic analysis.

Amino Acid Sequence↗

PubMed1992

Effect of atrial natriuretic peptide on apomorphine-induced stereotyped cage-climbing behavior in mice.

In previous experiments, it was observed that rat atrial natriuretic peptide-(1-28) (ANP-(1-28)) participated in fear-induced learning and memory processes via dopaminergic and cholinergic mediation. Since cage-climbing behavior is described as a simple test for studying dopaminergic activity in the central nervous system, a systemic study was carried out with ANP-(1-28) in order to confirm or to exclude the possible involvement of dopamine in the ANP-induced action in the brain. The present study demonstrate that ANP-(1-28) facilitated cage-climbing behavior in mice in a dose-dependent manner. When combined with apomorphine, the peptide potentiated the effect of the dopamine agonist. The effect of ANP-(1-28) in combination with apomorphine could be antagonized by a selected dose of haloperidol. These data suggest that ANP might be regarded as a dopamine agonist-modulating agent and that a dopaminergic mechanism is a possible mode of action of ANP in the fear-induced learning studied earlier.

Animals↗

PubMed1992

Critical role of endogenous corticotropin-releasing factor (CRF) in the mediation of the behavioral action of cocaine in rats.

The role of endogenous CRF in the locomotor hyperactivity induced by cocaine was investigated by using the immunoneutralization of endogenous CRF and an antagonist of CRF-receptors (alpha-helical CRF9-41: alpha h-CRF) in rats. Different dilutions of anti-CRF antibody (1:5, 1:20, but not 1:100) injected intracerebroventricularly (i.c.v.) 24 hours before the cocaine treatment blocked the expression of locomotor hyperactivity. Pretreatment with different doses (0.001, 0.01, 0.1, 1.0 micrograms i.c.v.) of alpha h-CRF inhibited the locomotor hyperactivity induced by cocaine dose-dependently. Neither the immunoneutralization nor the receptor blockade for CRF changed the hyperactivity induced by another locomotor stimulant caffeine. These results serve as indirect in vivo evidence of the selective role of endogenous CRF in the cocaine-induced behavioral alterations. The findings have implications as concerns the possible role of CRF in human psychopathological changes induced by cocaine.

Animals↗

PubMed1992

Effect of atrial natriuretic peptide on dopa potentiation in mice.

The effect of rat atrial natriuretic peptide (ANP1-28) on the pargyline-DOPA potentiation test was studied following its administration into the lateral brain ventricle in mice. Thirty minutes after pargyline pretreatment, three doses of ANP (200, 500, or 1,000 ng/mouse) were administered simultaneously with DOPA and animals were then observed for 2 h. ANP in doses of 500 and 1,000 ng markedly enhanced the effect of DOPA. The maximum intensity of the effect was registered 30-45 min following administration of the peptide. The data suggest that ANP might be regarded as a dopaminergic-modulating agent in the CNS.

Animals↗

PubMed1992

Regulation of transendothelial transport in the cerebral microvessels: the role of second messengers-generating systems.

Different elements of the intracellular signaling messenger systems have been detected in the course of our studies in the cerebral endothelial cells. It has been shown that the synthesizing enzymes of and substrate proteins for the second messenger molecules are present in the cerebral endothelial cells, and their activity and/or amount can change in pathological circumstances, i.e., during the formation of brain oedema. Pharmacological treatments interfering with the second messenger systems proved to be effective in the prevention of brain oedema formation.

Adenylyl Cyclases↗

PubMed1992

Behavioral effects of atrial and brain natriuretic peptides in rats.

The effects of intracerebroventricular administration of rat atrial natriuretic peptide (rANP-1-28) and porcine brain natriuretic peptide-32 (pBNP-32) on passive and active avoidance behavior and on electroconvulsive shock-induced amnesia were studied in rats. The dose range for both peptides was selected to lie between 0.016 and 0.32 nmol. The two peptides were found to facilitate consolidation of the passive avoidance response, to delay extinction of the active avoidance response, and to prevent electroconvulsive shock-induced amnesia in a similar way. It is suggested that some modulatory functions in the central nervous system of the rat, so far attributed to ANP, may in fact involve a dual control by both ANP and BNP, and there is no difference in the biological activity of the two peptides as far as fear-motivated learning behavior is concerned.

Amnesia↗

PubMed1991

The effects of atrial natriuretic peptide on active avoidance behavior in rats. The role of transmitters.

Different doses of rat atrial natriuretic peptide (rANP1-28) were tested as regards the extinction of active avoidance behavior following its injection into the lateral brain ventricle in rats. ANP delayed extinction of the active avoidance reflex in a dose-dependent manner. When the animals were pretreated with different receptor blockers in doses which themselves had no action on the extinction of active avoidance behavior, the action of ANP on this paradigm was completely blocked by haloperidol and atropine, whereas phenoxybenzamine/propranolol, naloxone, bicuculline and methysergide were ineffective. The data suggest that ANP delays the extinction of active avoidance behavior, and that cholinergic and dopaminergic transmitter systems might be involved in this action.

Animals↗