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Biomedical subjects

K Mann

Publications and source records attributed to K Mann.

At least 307 records · Page 17Linked to original sources
PubMed1994

Systemic iodine absorption during endoscopic application of radiographic contrast agents for endoscopic retrograde cholangiopancreaticography.

Hyperthyroidism induced by contrast agents in a major problem in patients with pre-existing thyroid disease, particularly in patients with functional thyroid autonomy. The present study was undertaken to evaluate whether contrast media applied during endoscopic retrograde cholangiopancreaticography (ERCP) may result in a significant increase of serum iodine levels and thus may be associated with the risk of iodine-induced hyperthyroidism. The courses of serum concentrations of total iodine and free iodide, as well as of urinary iodine excretion, were measured in 15 patients before and up to 21 days after ERCP. During ERCP, the non-ionic contrast medium iopamidol was instilled in amounts resulting in a total iodine load of 57.4 +/- 22.8 mmol (7.3 +/- 2.9 g). In all patients, ERCP resulted in a highly significant increase in serum levels of total iodine from 0.8 +/- 0.5 to 85.2 +/- 116.9 mumol/l 4 h after application of the contrast agent. In parallel, serum iodide levels were raised from 0.06 +/- 0.04 to 5.42 +/- 6.09 mumol/l and urinary iodine excretion from 71.1 +/- 35.7 mumol/mol creatinine to 621,620.9 +/- 636,492.2 mumol/mol creatinine. Peak concentrations of serum iodine are well related to the total amount of iodine applied (p < 0.05). During follow-up, iodine levels returned to pre-exposure levels within 2-3 weeks. Levels of thyrotropin, free thyroxine, and free triiodothyronine remained unchanged during the follow-up period. In conclusion, endoscopic application of iodinated contrast agents during ERCP leads to significant increases of serum levels of total iodine and free iodide and of urinary iodine excretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Absorption↗
PubMed1994

The molecular basis for epitopes on the free beta-subunit of human chorionic gonadotrophin (hCG), its carboxyl-terminal peptide and the hCG beta-core fragment.

The molecular basis for antigenic determinants on the free beta-subunit of human chorionic gonadotrophin (hCG beta), its carboxyl-terminal peptide (hCG beta CTP) and the hCG beta-core fragment (hCG beta cf) was elucidated by means of monoclonal antibodies (MCAs). The objective of the present study was to resolve the antigenic topography of these three molecules in terms of epitope identification at different levels of structural organization as well as analysis of their spatial arrangement. An hCG beta cf preparation, a synthetic peptide corresponding to the hCG beta CTP (beta 109-145), overlapping synthetic peptides spanning the entire amino acid sequence of hCG beta, and a reduced and alkylated hCG beta preparation were assayed in a solid-phase one-site enzyme-linked immunoassay and in a soluble-phase direct-binding radioimmunoassay (RIA) or competitive RIA. The antigenic topography was mapped by incorporating the MCAs into two-site binding assays. On the surface of free hCG beta, nine different epitopes (beta 1-beta 9), arranged in three spatially distinct domains, could be distinguished. Epitopes beta 1-beta 7 were located in a single large domain on both hCG beta and the hCG beta cf whereas hCG beta CTP contained two topographically distant determinants, designated beta 8 and beta 9 respectively. All but the two epitopes located on hCG beta CTP (beta 8 and beta 9) were destroyed by reducing and alkylating hCG beta, suggesting that most antigenic determinants are predominantly non-contiguous and require an intact tertiary structure whereas the molecular structure of hCG beta CTP is linear. At a molecular level, amino acid residues spanning hCG beta 45-52, hCG beta 137-144 and hCG beta 113-116 contributed to the formation of epitopes beta 5, beta 8 and beta 9 respectively. We have also shown that the hCG beta cf represents the immunodominant part of the free beta-subunit of hCG, containing seven mainly conformationally determined epitopes, one of which has a share of the sequence beta 45-52. The hCG beta CTP does not play a critical role in the immunologically important tertiary structure of hCG beta and was itself found to be a predominantly continuous sequence also within the native hormone, expressing two spatially distant antigenic determinants located within residues beta 113-116 and beta 137-144 respectively.

Antibodies, Monoclonal↗
PubMed1994

Nonlinear EEG dynamics during sleep in depression and schizophrenia.

The article deals with the question, whether nonlinear analysis of the sleep EEG may give clues to understanding of disturbed information processing in psychiatric diseases. We point out how the nonlinear approach to EEG dynamics is motivated and review recent investigations concerning nonlinear dynamical properties of physiological systems. We present calculations of the correlation dimension D2 and the principal Lyapunov-exponent lambda 1 for sleep EEG data from 9 depressive and 11 schizophrenic inpatients compared to healthy controls. Combining the findings for D2 and lambda 1 our results point to altered nonlinear brain dynamics mainly during slow wave sleep in depression and during REM sleep in schizophrenia. Finally, possible interpretations of altered nonlinear EEG dynamics during sleep concerning disturbed information processing in depression and schizophrenia are discussed.

Adult↗
PubMed1994

Immunological recognition and clinical significance of nicked human chorionic gonadotropin in testicular cancer.

We studied the physical properties, immunological recognition, and clinical significance of nicked human chorionic gonadotropin (hCGn) in testicular cancer. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions, the beta-subunit of hCGn (hCG beta n) dissociated into two peptides, and, by reversed-phase chromatography, hCG beta n was found to be less hydrophobic than hCG beta. Immunologically, hCGn lacked two epitopes specific for holo-hCG (c1, c2), whereas at least five hCG beta epitopes (beta 1-beta 5) were preserved, and, as a result, recognition of hCGn by different hCG assays varied widely. In 309 sera and 88 urine samples from patients with seminomatous or nonseminomatous testicular cancer, hCG-only, hCG+hCGn, and hCG+hCGn+hCG beta+hCG beta n+hCG beta core-fragment assays gave parallel results. hCGn was more abundant in urine than in serum samples. In conclusion, hCGn lacks two conformationally dependent epitopes of hCG, causing a change in hydrophobicity and explaining its failure to react in certain holo-hCG assays. Recognition of hCGn, however, does not seem to be crucial in the routine use of serum hCG as a tumor marker in patients with testicular cancer.

Chemical Phenomena↗
PubMed1994

Detection and possible functions of a cysteine protease involved in digestion of rod outer segments by retinal pigment epithelial cells.

PURPOSE: To investigate the presence and role of a recently cloned cysteine protease (cathepsin S) in the digestion of rod outer segments (ROS) by cultured retinal pigment epithelial (RPE) cells. METHODS: RPE cell cultures were established from eye bank donor eyes. Total RNA was extracted from freshly harvested cultures, and after reverse transcription, the cDNA was subjected to polymerase chain reaction (PCR). Cathepsin S (Cat S) mRNA translation was inhibited by antisense oligonucleotides, and the effect of inhibition on the accumulation of fluorescent debris was examined. The activity of cysteine and aspartic proteases in ROS-challenged RPE cell cultures was inhibited by leupeptin and pepstatin, respectively. The accumulation of autofluorescent debris within RPE cells was measured by a fluorophotometric flow cytometer. The presence of phagosomes in antisense DNA-inhibited and control cultures was demonstrated by electron microscopy. RESULTS: The expression of Cat S in RPE cells was demonstrated by RNA-PCR. Using antisense oligonucleotide-mediated-specific inhibition of Cat S, a significant ROS-derived increase in autofluorescence was detected within the RPE cells when they were compared with the unchallenged control cultures and cultures in the presence of ROS and sense oligonucleotides. Electron microscopy demonstrated the presence of a large number of phagosomes that enveloped structures similar to ROS. The accumulation of autofluorescent debris was also demonstrated in cysteine protease-inhibited, ROS-challenged RPE cultures, but it was not detected with aspartic protease inhibition. CONCLUSIONS: The expression of Cat S in RPE cells and the accumulation of an autofluorescent debris in cultures in which cysteine proteases or Cat S activity is inhibited suggest a key role for this enzyme, either in the ROS digestion process or in the activation of cathepsin D, the major lysosomal enzyme present in RPE cells.

Animals↗
PubMed1993

Characterization of two glycosylated boar spermadhesins.

Boar spermadhesins AQN-1, AQN-3 and AWN form a recently described protein family, synthesized by the sexual accessory glands, and become associated with the sperm head upon ejaculation. They contain 109-133 amino acid residues, two conserved disulphide bridges, are not glycosylated, and have 40-60% primary structure identity. These boar polypeptides are multifunctional proteins, which possess heparin-, serine-protease-inhibitor- and/or zona-pellucida-glycoprotein-binding capability and have, therefore, been implicated in sperm capacitation and sperm-oocyte attachment. AQN-2 (18-20 kDa), however, is unique among boar spermadhesins in that it is the only member of the family which is known to be glycosylated and which possesses weak zona-pellucida-binding but not seminal-plasma-inhibitor-binding ability. In this study we report the structural and functional characterization of the two glycoproteins contained in the AQN-2 fraction. One component is identical with PSP-I, a major porcine seminal plasma protein whose function has not yet been identified, while the second protein is a glycosylated isoform of AQN-3. Here we show that the inability of the glycosylated boar spermadhesins to bind seminal-plasma protease inhibitors as well as the weak binding of glycosylated AQN-3 to zona pellucida glycoproteins is due to the presence of the oligosacharide chain on a conserved asparagine residue. This indicates that modification of a spermadhesin polypeptide framework may serve to modulate its ligand-binding capabilities.

Amino Acid Sequence↗
PubMed1993

Sites of nidogen cleavage by proteases involved in tissue homeostasis and remodelling.

The cleavage of recombinant mouse nidogen in its native form was examined with granule-stored proteases (leucocyte elastase, mast-cell chymase), blood proteases (thrombin, plasmin, kallikrein), matrix metalloproteinases (stromelysin, matrilysin, collagenases) and, for comparison, with trypsin and the endoproteinase Glu-C. More than 50 major cleavage sites were identified by Edman degradation of several large fragments and smaller peptides. The data show an almost exclusive localization of protease-sensitive sites to the flexible segment, connecting the N-terminal globular domains G1 and G2, and within the C-terminal, laminin-binding domain G3. Domains G1, G2 and the rod-like segment were much more stable against proteolysis. Kinetic analysis indicated a fast cleavage of several different sites in the link region followed by destruction of G3 but this was to some extent variable depending on the particular protease. Leucocyte elastase was identified as the most active protease in the cleavage of nidogen whilst stromelysin, matrilysin, plasmin and kallikrein were of distinctly lower activity. No cleavage could be detected with interstitial collagenase and gelatinase A. The peptide analyses also allowed the location of two disulfide bridges within the G3 domain. Complex formation between nidogen and laminin fragments caused some protection against cleavage by thrombin, leucocyte elastase and stromelysin particularly in domain G3. The data indicate a relatively uniform cleavage pattern of nidogen which may be relevant in the context of protein/ligand-binding activities associated with domains G2 and G3. The proteolytic processes involved in remodelling and the cellular penetration of basement membranes could therefore be essential for the modulation of the mediator function of nidogen.

Animals↗
PubMed1993

Identification of a cross-linked double-peptide from the catalytic site of the Ca(2+)-ATPase of sarcoplasmic reticulum formed by the Ca(2+)- and pH-dependent reaction with ATP gamma P-imidazolidate.

The Ca(2+)-ATPase from sarcoplasmic reticulum can be inhibited by the Ca(2+)- and pH-dependent reaction with ATP gamma P-imidazolidate. The chemically and monofunctionally activated inhibitor introduces an intramolecular cross-link between two neighbouring peptides of the active site. This can be followed by the reduced mobility of the ATPase upon SDS-PAGE analysis which becomes even more pronounced after limited trypsinolysis. After cleavage of the cross-linked ATPase molecule by cyanogen bromide and separation of the peptides a double-peptide can be detected which upon sequencing can be identified as part of the phosphorylation and the nucleotide binding site, respectively.

Adenosine Triphosphate↗
PubMed1993

Mapping of nidogen binding sites for collagen type IV, heparan sulfate proteoglycan, and zinc.

Recombinant nidogen fragments comprising the globular domains G1 plus G2, the rod-like domain, and the rod connected to the globe G3 were prepared from the culture media of transfected human cell clones. In addition, domains G1 and G2 were separated from each other after cleavage with chymotrypsin. The purified fragments were characterized by N-terminal sequences, electrophoresis, electron microscopy, and radioimmunoassays and the cell clones by Northern hybridization. Transfection with a construct comprising a large part of domain G3 showed high mRNA levels but no secreted protein, indicating a protein folding problem. All these fragments were used as soluble and/or immobilized ligands in binding assays. This demonstrated major binding sites on domain G2 for collagen IV and heparan sulfate proteoglycan. Affinity chromatography on zinc- and cobalt-loaded columns showed binding of domains G2 and G3 and the rod. Protein binding, but not metal binding, was abolished by reduction and alkylation of nidogen. This allowed for the isolation of several zinc-binding tryptic peptides, four from G2, two from the rod, and one from the G3 domain. Most of these short peptides contained several histidines that are likely to mediate binding. Zinc inhibited efficiently G3-mediated nidogen binding to laminin at 4 degrees C (IC50 approximately 5 microM) but less at higher temperatures. Similarly, zinc inhibited binding to collagen IV and proteoglycan at low temperatures but not at high (37 degrees C) temperatures. This indicates a complex modulation of nidogen binding to other basement membrane proteins by some, but not all, transition metals. Whether the particularly striking effects shown for zinc are of biological relevance remains to be established.

Amino Acid Sequence↗
PubMed1993

Topoisomerase activity is associated with purified SV40 T antigen.

Purified SV40 T antigen has been assayed for topoisomerase activity. The ability to relax negatively-supercoiled SV40 DNA was found in preparations of T antigen purified either from human 293 cells infected with Ad5-SVR111 virus or from insect Sf9 cells infected with recombinant baculovirus 941T. The T antigen-associated relaxing activity was stimulated by MgCl2 and was not dependent on ATP, suggesting that it is not due to cellular topoisomerase II. The topoisomerase activity was immunoprecipitated by a monoclonal antibody specific for T antigen, but not by a control monoclonal antibody. In addition, immunoblotting of purified T antigen from human 293 cells with antihuman topoisomerase I and anti-human topoisomerase II antibodies failed to detect cellular topoisomerases I or II. Sedimentation analysis of purified T antigen revealed that the topoisomerase activity co-sedimented with the hexameric form of T antigen at 23S. The topoisomerase activity is, therefore, either inherent to T antigen or due to a cellular topoisomerase I tightly bound to, and co-purifying with, T antigen.

Animals↗
PubMed1993

A single EGF-like motif of laminin is responsible for high affinity nidogen binding.

A major nidogen binding site of mouse laminin was previously localized to about three EGF-like repeats (Nos 3-5) of its B2 chain domain III [M. Gerl et al. (1991) Eur. J. Biochem., 202, 167]. The corresponding cDNA was amplified by polymerase chain reaction and inserted into a eukaryotic expression vector tagged with a signal peptide. Stably transfected human kidney cell clones were shown to process and secrete the resulting fragment B2III3-5 in substantial quantities. It possessed high binding activity for recombinant nidogen in ligand assays, with an affinity comparable with that of authentic laminin fragments. In addition, complexes of B2III3-5 and nidogen could be efficiently converted into a covalent complex by cross-linking reagents. Proteolytic degradation of the covalent complex demonstrated the association of B2III3-5 with a approximately 80 residue segment of nidogen domain G3 to which laminin binding has previously been attributed. The correct formation of most of the 12 disulfide bridges in B2III3-5 was indicated from its protease resistance and the complete loss of cross-reacting epitopes as well as of nidogen-binding activity after reduction and alkylation. Smaller fragments were prepared by the same recombinant procedure and showed that combinations of EGF-like repeats 3-4 and 4-5 and the single repeat 4 but not repeats 3 or 5 possess full nidogen-binding activity. This identifies repeat 4 as the only binding structure. The sequence of repeat 4 is well conserved in the human and in part in the Drosophila laminin B2 chain.(ABSTRACT TRUNCATED AT 250 WORDS)

Amino Acid Sequence↗
PubMed1993

The alpha 1 beta 1 integrin recognition site of the basement membrane collagen molecule [alpha 1(IV)]2 alpha 2(IV).

Cells interact with type IV collagen mainly via the integrins alpha 1 beta 1 and alpha 2 beta 1. A triple helical CNBr derived fragment CB3[IV], which contains the recognition sites for both integrins, was isolated from type IV collagen. Trypsin treatment of CB3[IV] gave rise to four smaller fragments, F1-F4, of which the smallest one, F4, contained the recognition site for alpha 1 beta 1. Further fragmentation of F4 by thermolysin treatment at 50 degrees C led to fragment TL1, which represents the C-terminal half of F4, and which was no longer able to interact with alpha 1 beta 1. Therefore the recognition site of alpha 1 beta 1 had to be located within the N-terminal half of F4, a position which was verified by electron micrographs of a crosslinked F2-alpha 1 beta 1 complex. Modification of the Arg and Asp residues, which abolished the binding activity of F4, led to the identification of Arg (461) within the alpha 2(IV) and Asp (461) within the alpha 1 (IV) chain as essential residues for the alpha 1 beta 1. The array of these two residues on the surface of the triple helix is discussed.

Amino Acid Sequence↗
PubMed1993

Isolation and biochemical characterization of heparin-binding proteins from boar seminal plasma: a dual role for spermadhesins in fertilization.

Sperm surface-coated heparin-binding proteins originating from secretions of the male sexual accessory glands, are known to play a pivotal role as extrinsic regulatory factors during sperm capacitation in many mammalian species. They interact with glycosaminoglycans present in the female genital tract and enhance the subsequent zona pellucida-induced acrosome reaction. We have isolated heparin-binding proteins from boar seminal plasma by affinity chromatography on heparin-Sepharose and reverse-phase HPLC. N-Terminal sequence analysis of these proteins identified a boar counterpart of the bovine capacitation factors BSP-A1/2 (also called PDC-109) and BSP-A3. Several carbohydrate- and zona pellucida-binding proteins, which belong to the newly described spermadhesin family, were also identified as heparin-binding proteins. Our results imply that, besides other capacitation factors, members of the spermadhesin family may play a dual role in sperm capacitation and fertilization in the pig.

Amino Acid Sequence↗
PubMed1993

Increased binding capacity of receptors for the epidermal growth factor in benign thyroid nodules and thyroid malignancies.

To determine the role of epidermal growth factor (EGF) receptors in thyroid tumorigenesis, EGF binding was compared in membranes from malignant and from benign thyroid tumors. Surgical specimens were obtained from 28 patients with thyroid carcinomas (3 papillary, 13 follicular, 6 undifferentiated, and 6 medullary carcinomas) and from 30 patients with benign thyroid tumors (15 scintigraphically functional and 15 nonfunctional nodules). In 30 cases normal tissue adjacent to the tumor was also obtained. EGF binding was seen to be increased not only in thyroid carcinomas but also in benign thyroid tumors, particularly in functional thyroid adenomas. The highest EGF binding was found in undifferentiated carcinomas. A direct comparison of the EGF binding characteristics in tumor and adjacent normal thyroid tissue revealed that the increased binding of EGF is due mainly to an increase in the number of binding sites rather than an alteration in receptor affinities. EGF binding capacities were 18.4 +/- 16.7 fmol/mg protein in thyroid carcinomas and 10.5 +/- 5.2 fmol/mg in the corresponding normal tissue (P < 0.05, Kd 0.84 +/- 0.26 nM, n = 11). In autonomously functioning thyroid adenomas binding capacities were 14.2 +/- 8.2 fmol/mg in the nodules and 8.9 +/- 4.8 fmol/mg in normal tissue (P < 0.01, Kd 0.73 +/- 0.62 nM, n = 15). In conclusion, EGF receptor levels are increased not only in malignant thyroid tumors but also in well-differentiated benign thyroid nodules.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1993

Reversible hyperkinesia in a patient with autoimmune polyglandular syndrome type I.

Autoimmune polyglandular syndrome is characterized by a failure of multiple endocrine organs and the presence of circulating organ-specific autoantibodies targeted against the failing organs. Here we describe a patient with autoimmune polyglandular syndrome type I with the endocrine manifestations of hypoparathyroidism, adrenocortical insufficiency, and insulin-dependent diabetes mellitus. Long-standing hypoparathyroidism led to extensive calcification of the basal ganglia which resulted in the clinical presentation of an extrapyramidal movement disorder (choreoathetotic and hemiballistic hyperkinesia of the left extremities). Interestingly, parallel to rehydration and the initiation of cortisol replacement therapy a complete reversion of the hyperkinetic signs was achieved. This case shows a rare multiendocrine organ failure with complex metabolic interactions resulting in marked neurological signs. Furthermore, this case demonstrates for the first time that a hyperkinetic syndrome--most likely due to hypoparathyroidism-induced basal ganglia calcification--can be reversed solely by adequate treatment of the concomitant endocrine failures.

Adrenal Cortex Diseases↗
PubMed1993

Molecular heterogeneity of human chorionic gonadotropin in serum and urine from patients with trophoblastic tumors.

Many studies on the structure and function of human chorionic gonadotropin (hCG) have relied on purified hCG preparations obtained from pregnancy urine. In the present studies, in order to demonstrate possible differences between the hCG species present in serum and those released into urine, we examined serum and urinary samples derived from patients with trophoblastic tumors and of pregnancy origin by sodium dodecyl sulfate electrophoresis, isoelectric focusing, and two-dimensional electrophoresis including immunostaining with a specific hCG antibody and densitometry of the protein bands. This type of analysis was presently feasible only in patients presenting with extremely high serum levels of hCG and was therefore limited to seven patients with testicular cancers, one woman with a hydatidiform mole, and one pregnancy sample. We found marked differences in the isoelectric focusing pattern between urinary and serum hCG samples, with the urinary hCG consisting of more alkaline pI variants than that present in the serum of the same patients. Tumor hCG differed from pregnancy hCG in that it contained more acidic variants, and this was true for urinary and serum-derived materials. In some tumor and pregnancy samples an hCG immunoreactive material of lower molecular weight than hCG itself was found. In conclusion, the present studies, extending previous findings on the microheterogeneity of hCG, indicate that serum and urinary-derived hCG may differ in the composition of the isoform spectrum, as does tumor and pregnancy hCG. Further, in some patients hCG immunoreactive molecules exist that differ markedly from hCG in size and charge. These observations suggest that, whenever possible, serum-derived hCG materials should be used to define the molecular structure of hCG and assess its biological activities.

Adult↗
PubMed1993

Self-regulation of slow cortical potentials in psychiatric patients: alcohol dependency.

Ten unmediated alcohol-dependent male inpatients participated in a Slow Cortical Potential (SCP) self-regulation task utilizing biofeedback and instrumental conditioning. These patients were hospitalized for treatment of alcohol dependency after chronic abuse of alcoholic beverages. Somatic withdrawal symptomatology had occurred recently and the patients were free of any withdrawal symptoms of the autonomic nervous system. Immediately after hospitalization patients were unable to control their SCPs without the reinforcement of immediate feedback across 4 sessions. Seven patients participated in a fifth session an average of 4 months later. Six out of these 7 patients had not had a relapse at the follow-up. In the fifth session these patients were immediately able to differentiate between the required negativity and negativity suppression, whereas the seventh patient, who had relapsed, was unable to control his brain potentials successfully. Results are further evidence that some of the frontocortical dysfunctions in alcohol-dependent patients are reversible. This could covary with a morphological restitution of the cortex.

Adult↗