Biomedical subjects

H Winkler

Publications and source records attributed to H Winkler.

At least 181 records · Page 10Linked to original sources

PubMed1990

Comparison between short-course high-dose preoperative irradiation with immediate radical cystectomy and salvage cystectomy after full-dose irradiation.

Between 1976 and 1983, 70 patients who underwent cystectomy for transitional cell carcinoma (TCC) of the bladder were retrospectively divided into two groups: 39 patients were treated by a protocol using 2,000 rad of radiation over a period of 5 days followed by immediate cystectomy (group 1); 31 patients who failed to be cured by definitive radiotherapy of 6,000 rad were treated by salvage cystectomy (group 2). The 5-year disease-specific survival rate was not significantly different in the two groups of patients (64.5 and 65.5%). The postoperative early complication rates were similar as well (36.8% in group 1 and 35.5% in group 2). One patient of the group treated with 2,000 rad died 19 days after surgery, giving an operative mortality of 1.4%. We conclude that there is a continuing role for salvage cystectomy after irradiation failure. One-stage salvage cystectomy has a relatively low morbidity and should be performed early after failure of definitive irradiation.

Aged↗

PubMed1990

Process analysis in behavioral family therapy.

Process research in behavioral family therapy (BFT) is a hitherto neglected area with the notable exception of the work of Alexander and Patterson with their colleagues. However, the development of instruments to assess therapist's behavior during the treatment session seems timely for the following reasons: (a) to investigate the relationship between therapist behavior and outcome, (b) to improve therapist training/supervision, (c) to establish treatment integrity in comparative outcome studies, and (d) to monitor treatment progress. For these purposes two sets of instruments were developed: (a) a category system to describe the content of a session, which is rated every 30 seconds and (b) several rating scales to evaluate therapist behavior, including relationship competency, ability to structure a session, didactic competence, ability to initiate behavioral probes, use of appropriate intervention strategies, use of reinforcement, and dealing with uncooperativeness. Scales are described, and data on their reliability and utility are presented.

Adult↗

PubMed1990

Chromogranins A and B are co-localized with atrial natriuretic peptides in secretory granules of rat heart.

We investigated the occurrence and subcellular localization of chromogranins A and B in atrial myoendocrine cells of rat heart, using immunological methods. Immunoblotting revealed the presence of both chromogranin A and B in an extract from large granules isolated from this tissue by subcellular fractionation. Immunohistochemistry at the ultrastructural level demonstrated the presence of chromogranin A and B in secretory granules. These organelles also immunostained for atrial natriuretic peptides (ANP). Within a given section, all granules were labeled with immunogold for these three antigens. This apparent co-localization of the three antigens was confirmed by double immunostaining with immunogold particles of different sizes. We conclude that, in agreement with their endocrine nature, the secretory organelles of rat atria contain both chromogranins A and B. Apparently these acidic peptides, which have a widespread distribution in the endocrine system, are co-stored and therefore also co-secreted with ANP.

Animals↗

PubMed1990

[Eruptive viral warts of the burn area].

Eruptive viral warts were observed in a second-to-third degree burn area in a 68-year-old woman; the warts healed spontaneously and completely after approximately 3 weeks. HPV-2-related DNA was demonstrated by Southern blot analysis.

Aged↗

PubMed1990

Chromogranin A and B gene expression in carcinomas of the breast. Correlation of immunocytochemical, immunoblot, and hybridization analyses.

Chromogranins (Cg) are regarded as specific neuroendocrine (NE) markers in cells and tumors. Expression of CgA and CgB genes has been demonstrated by correlative immunocytochemical, immunoblotting, in situ hybridization, and Northern blot procedures in seven argyrophilic breast carcinomas, while eight control cases of ductal carcinomas, not otherwise specified, were negative. A high degree of correlation was observed between the various techniques revealing CgA and/or CgB gene expression at different levels; minor discrepancies might be related to tumor heterogeneity or to technical factors. The present study, confirming previous investigations, establishes NE differentiation in a group of human breast cancers. The identification of this type of tumors, especially by testing chromogranin(s) production, appears to be of both biologic and clinical interest.

Biomarkers, Tumor↗

PubMed1990

[Value of functional treatment of severe calcaneus fractures].

Preservation of the gliding joint and muscular strength, avoidance of contracture, and improvement of gait pattern are the major objectives in treatment of patients for severe calcaneal fracture. Follow-up checks were applied to 170 patients, with particular reference being made to the value of functional therapy. Unobstructed mobility of the talocrural joint was recorded from 90 percent of them. Occupational fitness was reduced in 25 percent of the above cases.

Adolescent↗

PubMed1990

[Experiences with intraluminal small intestinal intubation].

Seventy-six patients were treated for postoperative ileus of the small intestine, between 1983 and 1987. Forty-four of them received intraluminal intestinal intubation, with indications being established by stringent criteria. One case of recurrent ileus was recorded eight weeks after removal of the Miller-Abbott tube, but no tube-related lethality was observed. The average age of our patients was as low as 49 years. Post-operative lethality amounted to 21.2 percent (eight in 44). Ileus was not removed until death in three cases. Intraluminal intestinal intubation may be recommended after long-distance lysis of adhesions for postoperative ileus of the small intestine as well as in cases of severe ileus in concomitance with controllable peritonitis.

Aged↗

PubMed1989

Adrenal chromaffin granules and secretory granules from thyroid parafollicular cells have several common antigens.

The presence of various antigens in two types of isolated endocrine vesicles (chromaffin granules and secretory vesicles of thyroid parafollicular cells) was investigated by immunoblotting. The two types of vesicles have three common secretory proteins: chromogranin A, chromogranin B and secretogranin II. Furthermore, six common membrane antigens were found: cytochrome b-561, carboxypeptidase H, glycoprotein II, glycoprotein III, synaptin/synaptophysin and SV 2. These results demonstrate that vesicles obtained from neural crest-derived endocrine cells not only share several common secretory peptides and proteins, but also have common properties as far as their membrane antigens are concerned.

Adrenal Medulla↗

PubMed1989

Fe(II)-substituted horse liver alcohol dehydrogenase, a model for non-heme iron enzymes. Various states of iron-dioxygen interaction investigated by Mössbauer and EPR spectroscopy.

The catalytic Zn(II) ion of horse liver alcohol dehydrogenase (EE isozyme) was replaced by Fe(II), providing a novel iron protein with the unusual coordination of Fe(II) to two cysteines residues, one histine residue and water. The electronic structure of iron in this system was characterized by Mössbauer spectroscopy at various temperatures as well as applied magnetic fields and analysed in terms of the spin Hamiltonian formalism. The novelty we found is an unusually weak spin coupling (/J/ less than 0.1 cm-1) of a paramagnet (S = 1) with iron (S = 2). From EPR and biochemical studies we conclude that the corresponding chemical species is triplet oxygen (O2). The quantitative determination of the coupling energy was possible utilizing the competition between Zeemann interaction and spin coupling at weak magnetic fields and low temperature. Oxidation experiments followed by Mössbauer spectroscopy showed that the spin-coupled system is an outer-sphere Fe(II) . (O2)aq complex occurring as an intermediate during a Fe(II)-catalyzed dioxygen activation. We observed two additional Fe(II) species after treatment with O2 and dithionite. The spin Hamiltonian parameters of iron in the coupled system are presented. The results are compared with those of iron in other non-heme iron proteins.

Alcohol Dehydrogenase↗

PubMed1989

Ontogenesis of chromogranin A and B and catecholamines in rat adrenal medulla.

The ontogenesis of chromogranins A and B and catecholamines was investigated for rat adrenal medulla. The chromogranins, the major secretory peptides of chromaffin granules, were characterized by one- and two-dimensional immunoblotting. Chromogranin B appeared identical in fetal and postnatal adrenals. On the other hand a significant portion of chromogranin A immunoreactivity in fetal adrenals was present in a component which was identified as a proteoglycan form of chromogranin A. In adult adrenals this proteoglycan chromogranin A was different and much less prominent. Total chromogranins and catecholamines increased parallely from the 17th prenatal day to the adult stage. However in early periods of development chromaffin granules are likely to contain relatively higher catecholamine levels than adult granules. At the end of the gestational period both the ratio of adrenaline to noradrenaline and of chromogranin A vs B increases sharply. These results establish that from the 17th prenatal day onward fetal adrenal glands contain chromaffin granules which are filled with both the major secretory peptides and catecholamines. The relative composition of the secretory content (adrenaline vs noradrenaline and chromogranin A vs chromogranin B) changes at the end of the gestational period in parallel with the development of the corticosteroid-producing adrenal cortex.

Adrenal Medulla↗

PubMed1989

Erythrocyte pyruvate kinase deficiency: a kinetic method for differentiation between heterozygosity and compound-heterozygosity.

The goal of the present study was to search for criteria that allow one to distinguish between normal individuals and heterozygotes as well as compound heterozygotes for pyruvate kinase (PK) deficiency. As the residual activity of PK with heterozygotes was between 35% and 110% of the normal activity, it was necessary to find other methods to prove heterozygosity. The PK in the hemolysates of 23 patients suffering from PK deficiency, 36 paternal and maternal enzymes as well as the enzymes of five heterozygous and four normal siblings together with those of 20 normal individuals, were studied according to the recommendations of the International Committee for Standardization in Haematology. The following hematological and enzyme kinetic parameters can serve to identify heterozygotes for PK deficiency: 1) a slight reticulocytosis, 2) an up-to-twofold increase of the intracellular concentrations of glucose-6-phosphate in the erythrocyte, 3) a mixed cooperativity of the phosphoenolpyruvate (PEP)-binding process of PK, 4) a decreased nucleotide specificity with guanosine diphosphate and uridine diphosphate, and 5) a lowered affinity for adenosine diphosphate. The most significant criterium found with all heterozygotes was a mixed cooperativity of the PEP-binding process caused by the presence of a mixture of normal and mutant PK.

Adolescent↗

PubMed1989

Co-localization of chromogranin A and B, secretogranin II and neuropeptide Y in chromaffin granules of rat adrenal medulla studied by electron microscopic immunocytochemistry.

The co-localization of various antigens in rat chromaffin granules was investigated by the immunogold staining procedure. In ultrathin serial sections staining of chromaffin granules was obtained with antisera against chromogranin A, chromogranin B, secretogranin II and neuropeptide Y. These results indicated that these antigens are costored within chromaffin granules. To further corroborate this point a double immunogold staining procedure was used. This method unequivocally established that chromogranin A, chromogranin B, secretogranin II and neuropeptide Y are co-localized in the same chromaffin granules. These results are relevant for studies demonstrating changes in the level of these peptides in adrenal medulla. The co-localization makes it likely that such changes lead to a different relative composition of the secretory quanta of chromaffin granules.

Adrenal Medulla↗

PubMed1989

An immunological study on chromogranin A and B in human endocrine and nervous tissues.

Antisera were raised against synthetic peptides derived from the primary amino acid sequence of human chromogranin B. These antisera recognized in one- and two-dimensional immunoblotting a component previously designated as chromogranin B. In human chromaffin granules, the major endogenous processing product of chromogranin B is formed by proteolytic cleavage of the protein near the C-terminus. Immunohistochemical localizations were obtained with antisera against human chromogranins A and B and against a synthetic peptide corresponding to the B sequence. In human tissues, chromogranin B is co-stored with chromogranin A in the adrenal medulla, the anterior pituitary, parafollicular cells of the thyroid, in some cells of the endocrine pancreas and in some enterochromaffin cells, whereas only chromogranin A is found in the parathyroid gland and enterochromaffin cells of the gastric corpus mucosa. In the nervous system, no immunostaining was observed for chromogranin A and only a weak one for chromogranin B in some cells of the spinal cord. However, the Purkinje cells of the cerebellum were strongly positive for chromogranin B.

Chromogranin A↗

PubMed1989

Insulin hypoglycemia increases the levels of neuropeptide Y and calcitonin gene-related peptide, but not of chromogranins A and B, in rat chromaffin granules.

The levels and subcellular distribution of chromogranin A and B, of calcitonin gene-related peptide (CGRP) and of neuropeptide Y (NPY) were investigated in rat adrenals before and after insulin treatment. Six days after insulin-induced hypoglycemia the levels of chromogranin A and B were similar to controls, however those of NPY and CGRP were increased by a factor of 2.5 and 35, respectively. This treatment also elevated mRNA levels of NPY and CGRP, establishing an increased biosynthesis of these two neuropeptides. As shown by subcellular fractionation, all peptides were present in chromaffin granules after insulin treatment. Furthermore, immunostaining at the ultrastructural level demonstrated the co-localization of chromogranin A, NPY and CGRP within the same chromaffin granules. These results establish that insulin-induced hypoglycemia changes the levels of the secretory peptides in chromaffin granules leading to an altered composition of the secretory cocktail. Apparently, the biosynthesis of the secretory peptides and their storage organelles can be regulated in distinct patterns.

Animals↗

PubMed1989

Influence of the red blood cell Ca2+-ion concentration on the erythrocyte aggregation in stasis.

Ca2+ ions were transported into the cell by incubation of the erythrocyte suspension with ionophore A23187, a lipophil electric neutral ion complexing substance. Erythrocyte aggregation could be increased twice, when doubling the intracellular Ca2+-ion concentration. Our measurements lead to the suggestion that an increase of the cytoplasmatic Ca2+ ion changes the physical and/or biochemical properties of the aggregation receptors on the membrane surface, i.e., cell-protein interactions are regulated by alteration of the intracellular Ca2+-ion concentration.

Adolescent↗

PubMed1989

On the diagnosis of erythrocyte enzyme defects in the presence of high reticulocyte counts.

The separation of red blood cells into reticulocytes and young and old erythrocytes enables investigations of fractions with different contents of reticulocytes. Activities of hexokinase, glucose phosphate isomerase, phosphofructokinase, pyruvate kinase and glucose-6-phosphate dehydrogenase showed a linear relationship to reticulocyte counts. The dependence of these enzyme activities on the age of the red blood cells exhibited a strong decline from the reticulocyte to the young erythrocyte stage followed by only little further loss of activity, thus leading to a biphasic decay of enzyme activities. By linear regression analysis enzyme activities in erythrocytes (AE) and reticulocytes (AR) could be evaluated. The activity of a given enzyme in the reticulocyte exceeded that of the erythrocyte; the quotient AR/AE represents the decline of enzyme activity from the reticulocyte to the erythrocyte stage. This value AR/AE is 16.7 for pyruvate kinase and 9.4 for hexokinase and thus considerably higher than that for the other enzymes investigated (glucose phosphate isomerase: 2.9, phosphofructokinase: 4.3, glucose-6-phosphate dehydrogenase: 4.5). In patients suffering from erythrocyte enzymopathies, the AR/AE for pyruvate kinase was 16.2 and thus almost identical to the normal enzyme. Calibration curves where the enzyme activity is plotted versus the fraction of reticulocytes enable the determination of normal activity of a given erythrocyte enzyme depending on the content of reticulocytes in red blood cell suspensions. Thus an unambiguous diagnosis of enzyme defects irrespective of reticulocyte counts becomes possible.

Adult↗

PubMed1989

Calcium content of the erythrocytes: a sensitive and easy handling method for measuring free calcium ions, and modulation of the Ca2+ ion concentration by the calcium antagonists nifedipine and pentoxifylline.

1. A method for determining free Ca2+-ions in the erythrocyte is described, using a commercially available ORION-Ca-electrode and calomel reference electrode assembly, where changes in free Ca2+-ion concentration upon addition of 0.01% digitonin could be measured. 2. The average value found for fresh cells from 20 healthy donors at 37 degrees C (pH = 7.4) was 0.20 +/- 0.04 mumol/L referred to a haematocrit of 10%. 3. Decrease of the simultaneously determined adenosinetriphosphate (ATP) concentration indicates that ATP is presumably needed to activate the Ca-ATPase. 4. In vitro addition of the calcium antagonists pentoxifylline and nifedipine, respectively, induced a normalization of the intraerythrocytic Ca2+-ion concentration after previous increase with the ion carrier ionophore A23187. 5. The advantages and possible clinical applications of this method are discussed.

Adenosine Triphosphate↗

PubMed1989

An increased pool of secretory hormones and peptides in adrenal medulla of stroke-prone spontaneously hypertensive rats.

Secretory components of the adrenal medulla were compared in normotensive Wistar-Kyoto (WKY) rats and in stroke-prone spontaneously hypertensive rats (SHRSP) at both 4 and 12 months of age. Noradrenaline, adrenaline, dopamine, neuropeptide Y, and chromogranins A and B were significantly higher in adrenal glands of SHRSP than those of WKY rats at 4 months. At 12 months, the levels of these components in SHRSP had increased even more (about 200% in WKY rats). There was no change in the relative composition of the adrenal "secretory cocktail." Neither the chromogranin A/chromogranin B ratio nor their apparent proteolytic processing in chromaffin granules differed between SHRSP or WKY rats. The lack of a significant change in membrane-bound cytochrome b561 and the small increase in dopamine beta-hydroxylase suggest that the higher levels of secretory components in SHRSP are not simply caused by an increase in the number of chromaffin granules, but possibly by a selective increase in the secretory content of these organelles providing a larger package for quantal release by exocytosis. This may be relevant for the elevation of blood pressure in this strain. The immunological methods described in this paper allow for the first time a determination of the secretory quantal levels in catecholamine storage. This should be useful for further studies in hypertensive models.

Adrenal Medulla↗