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Biomedical subjects

K Weber

Publications and source records attributed to K Weber.

At least 181 records · Page 10Linked to original sources
PubMed1996

Normal respiratory chain function in patients with low-tension glaucoma.

OBJECTIVE: To test the hypothesis that low-tension glaucoma has a pathogenesis similar to Leber's hereditary optic neuropathy and results from a defect in the mitochondrial respiratory chain. METHODS: Mitochondrial fractions were prepared from skeletal muscle samples collected from eight subjects with low-tension glaucoma. Their oxidative metabolism was compared with that of age- and sex-matched controls. Skeletal muscle DNA prepared from the subjects with glaucoma was also screened for the 3,460, 11,778, and 14,484 mitochondrial DNA mutations that are associated with Leber's hereditary optic neuropathy. RESULTS: No subject with low-tension glaucoma had a defect in respiratory chain activity or one of three mitochondrial DNA mutations that are commonly associated with Leber's hereditary optic neuropathy. CONCLUSION: Although these results do not exclude the possibility that low-tension glaucoma is caused by an organ-specific defect of mitochondrial function, we have excluded a systemic defect of the mitochondrial respiratory chain.

Adult↗
PubMed1996

Comparison of the hemodynamic effects of metoprolol and carvedilol in hypertensive patients.

Metoprolol and carvedilol are widely used in the treatment of hypertension, but no randomized comparison of their hemodynamic activity has been previously reported. Their comparative effects on heart rate, systemic blood pressure, and echocardiographically determined aortic and femoral artery blood flow were measured at rest and at 2 and 24 hours after the first dose of each drug, and again after 4 weeks of sustained monotherapy in 12 male and 12 female patients, aged 36-68 years with uncomplicated sustained hypertension according to a randomized single-blind protocol. Nine patients in each drug group achieved the target diastolic blood pressure of < 90 mmHg on the initial doses of each drug; this was achieved in the remainder following doubling of each dose. Neither drug occasioned withdrawal of any patient due to adverse reactions. Both drugs significantly reduced heart rate, although the reduction at 2 hours was significantly greater after metoprolol than after carvedilol. Both drugs reduced systolic pressure throughout the study; the reduction at 2 hours was significantly greater after carvedilol than after metoprolol. In contrast, the diastolic blood pressure was persistently reduced only by carvedilol. The cardiac output, determined as the aortic systolic blood flow, after carvedilol was not significantly different from pretreatment values throughout the study but was significantly reduced in the metoprolol-treated patients at each point of measurement. After metoprolol the systemic and femoral vascular resistances derived from conventional formulae were consistently and significantly increased over pretreatment values throughout the study and were significantly greater than in the carvedilol group at all measurement points. The hemodynamic differences between these two beta-blocking drugs may be explained by the additional vasodilator activity of carvedilol associated with its alpha 1-adrenoceptor blocking activity. The long-term clinical and prognostic implications of these pharmacodynamic differences between beta-adrenoceptor antagonists with and without additional vasodilator activity in the treatment of hypertensive patients remain to be determined.

Adrenergic alpha-Agonists↗
PubMed1996

Effect of O2 availability on neuroendocrine variables at rest and during exercise: O2 breathing increases plasma prolactin.

Neuroendrocrine and substrate responses were investigated in eight male athletes during inhalation of either 100% O2 (HE), 14% O2 (HO) or normoxio gas (NO) before, during and after 60 min of cycle ergometry at the same absolute work rate. Concentrations of prolactin (PRL), growth hormone (GH), testosterone (T), adrenocorticotropic hormone (ACTH), cortisol (COR), adrenalin (A), noradrenalin (NA), insulin (INS), ammonia (NH3), free fatty acids, serotonin (5-HT), total protein, branched-chain amino acids (BCAA) and free tryptophan (free TRP) were determined in venous blood and lactate concentration [LA-], partial pressure of oxygen (PO2), oxygen saturation (SO2), partial pressure of carbon dioxide and pH in capillary blood. The PO2 and SO2 were augmented in HE and decreased in HO (P < or = 0.01). In HO and NO no significant changes were found for any other parameter during 30 min of rest prior to exercise. In HE, PRL increased by about 400% during this time, while NA declined (P < or = 0.01). Heart rate (HR) and [LA-] were higher during exercise in HO (P < or = 0.01). In all trials, NH3, NA, A, T, GH and ACTH increased during exercise (P < or = 0.01), while BCAA and INS declined. In comparison to NO and HE, increases of NA, A, GH, COR and ACTH were higher in HO (P < or = 0.01). The PRL in NO and COR in NO and HE did not change significantly. In HE, after the initial increase at rest, PRL declined during exercise but remained higher than in HO. Higher values for NA, A, GH, COR and ACTH in HO were likely to have reflected an augmented relative exercise intensity. Our results showed that PRL but no other hormone increased during acute exposure to hyperoxia. This PRL release was independent of exercise stress and greater than PRL augmentation during hypoxia, which was related to a higher relative exercise intensity as indicated by [LA-] and HR. Responses of plasma NH3, BCAA, free TRP and 5-HT could not explain PRL augmentation induced by the increment in blood SO2 during hyperoxia.

Adrenocorticotropic Hormone↗
PubMed1996

[Value of ultrasound examination in injuries of the symphysis].

Lesions of the symphisis are usually diagnosed by a pelvis X-ray. The use of ultrasound examination is not very common. The application of the ultrasonic head in a suprasymphyseal manner will result in good images of the anterior pelvis. The outcome of diastasis as well as of the edge of the symphysis measurements taken by ultrasound were comparable to the X-ray picture. Therefore ultrasound investigation is usefull for primary diagnosis and follow-up examination in injuries of the symphysis. Accompanying fractures of the pelvic ring stay in this case undetected.

Adult↗
PubMed1996

Structural and functional characterization of human potassium channel subunit beta 1 (KCNA1B).

Voltage-activated Shaker-related potassium channels (kv1) consist of alpha and beta subunits. We have analysed the structure of the human KCNA1B (hKv beta 1) gene. KCNA1B is > 250 kb in size and encodes at least three Kv beta 1 splice variants. The Kv beta 1 open reading frame is divided into 14 exons. In contrast, genes coding for family members of KCNA (Kv 1 alpha) subunits are markedly smaller and have intronless open reading frames. The expression of Kv 1 alpha and Kv beta mRNA was compared in Northern blots of poly(A+) RNA isolated from various human brain tissues. The results suggest an intricate and cell-specific regulation of Kv 1 alpha and Kv beta mRNA synthesis such that distinct combinations of alpha and beta subunits would occur in different nuclei of the brain. The splice variants hKv beta 1.1 and hKv beta 1.2 were functionally characterized in coexpression studies with hKv 1.5 alpha subunits in 293 cells. It is shown that the confer rapid inactivation on hKv 1.5 channels with different potencies. This may be due to differences in their amino terminal sequences and/or inactivating domains. It is also shown that the amino terminal Kv beta 1.1 and Kv 1.4 alpha inactivating domains compete with each other, probably for the binding to the same receptor site(s) on Kv 1 alpha-subunits.

Amino Acid Sequence↗
PubMed1996

Increased prevalence of IgA-Chlamydia antibodies in NIDDM patients.

Chlamydia trachomatis oculogenital infection is a common disease in western societies. Despite the fact that diabetes is accompanied by increased risk for infections, no data on chlamydial infections in the non-insulin-dependent diabetic (NIDDM) patient exist. In our study Chlamydia antibodies were determined using an immunoperoxidase reaction in NIDDM patients (n = 79) and in a local nondiabetic control population (n = 125) which was randomly invited to a medical control visit without any preselection criteria. In total, 46% of diabetics and 55% of controls were IgG-Chlamydia antibody positive (ns). Using IgA-Chlamydia antibodies to define 'seroactive' chlamydial infection, 22% of NIDDM patients and 14% of controls were positive. Thus seroactive chlamydial infection of all patients with proven contact to Chlamydia (IgG-Chlamydia antibody positive) was 47% in diabetics versus 25% in controls, respectively (P < 0.05). Forming subgroups, significance was reached in females (52% vs. 32%, P < 0.05) only, but a similar trend was observed in males (36% vs. 21%, ns). Seroactivity was neither correlated with HbA1c nor with nephelometrically determined total serum immunoglobulins (IgG, IgA). Additionally we observed significantly elevated total IgM and IgA-levels in NIDDM patients whereas IgG-levels were comparable in both groups. In conclusion, seroactive chlamydial infections in subjects with proven contact to Chlamydia are more frequent in NIDDM patients than in nondiabetic controls. Additionally, higher IgM and IgA serum levels might indicate a higher susceptibility to active surface infections in NIDDM.

Adult↗
PubMed1996

On the nature of the irreversible inhibition of histidine ammonia lyase by cysteine and dioxygen.

The irreversible inhibition of histidine ammonia lyase by L-cysteine and dioxygen has been reexamined. After denaturation and consecutive digestion of the inhibited enzyme by trypsin and endoproteinase Glu-C, the generated chromophoric system (lambda max = 340 nm) remained intact and was isolated in an octapeptide containing amino acids 138-145, as previously described (Hernandez, D.; Stroh, J.G.; Phillips, A. T. Arch. Biochem. Biophys. 1993, 307, 126). Conducting the inhibition in the presence of 18O2 did not result in the incorporation of the heavy isotope into the isolated octapeptide. Total hydrolysis followed by amino acid analysis of the octapeptide revealed the presence of one arginine in addition to those expected from the deduced sequence (G3SVAD). 1H NMR spectroscopy of the octapeptide confirmed the presence of the amino acids GSVAD and showed no signals for olefinic or aromatic protons. To account for the excess mass of the octapeptide, we propose an oxidative degradation of the dehydroalanine prosthetic group, followed by reaction of the resulting dicarbonyl system with a nearby arginine residue.

Chromatography, High Pressure Liquid↗
PubMed1996

Feasibility of labeled alpha-acetamido-aminoisobutyric acid as new tracer compound for kinetic labeling of neutral amino acid transport: preparation of alpha-(N-[1-11C]acetyl)- and alpha-(N-[1-14C]acetyl)-aminoisobutyric acid.

The nonphysiological, nonracemic, branched-chain alpha-acetamido-aminoisobutyric acid was labeled with the carbon isotope 11C with the intention to use it in conjunction with positron emission tomography (PET) to measure the kinetics of amino acid transport in vivo. It was produced by the reaction of the novel 11C-precursor N-[1-11C]acetylpyridinium chloride with alpha-aminoisobutyric acid. Typically, 2 GBq of alpha-(N-[1-11C]acetyl)-aminoisobutyric acid were isolated with a specific activity of 12 to 20 GBq. mumol-1 at the time of application, and with a radiochemical purity of > 98%. The chemical identity of alpha-(N-[1-11C]acetyl)-aminoisobutyric acid was confirmed by comparison with alpha-(N-[1-14C]acetyl)-aminoisobutyric acid that was independently prepared by a standard acetylation procedure of alpha-aminoisobutyric acid using [1-14C]acetic anhydride. In vivo, both labeled substrates were not metabolized. In cell-culture experiments, 84% of the substrate entered the cells by the sodium-dependent amino acid transport system A, whereas 16% was taken up by the sodium-independent system. The uptake of the radiotracer was measured 20 min and 40 min postinjection in tumor-bearing male Copenhagen rats for assessment of its in vivo biodistribution.

Adenocarcinoma↗
PubMed1996

Alterations in plasma free tryptophan and large neutral amino acids do not affect perceived exertion and prolactin during 90 min of treadmill exercise.

It has been hypothesized that fatigue and prolactin (PRL) changes during endurance exercise are influenced by serotonin synthesis, and in turn, release. Such a change is thought to occur through an increase in blood free tryptophan (TRP) and a concomitant decrease in those large neutral amino acids (LNAA) which compete with free TRP for entry into the brain. For further investigation, 10 healthy athletes were randomly subjected to three test units (TU), each consisting of a treadmill run for 90 min. The speed was adjusted to a blood lactate level of 2 mmol/l. During the first 30 min of exercise infusions of 500 ml saline (TU I), 500 ml saline with amino acids (TU II) or 500 ml saline with 30 U heparin/kg following an oral soy oil solution given 1 h before (TU III) were administered. Rate of perceived exertion (RPE), heart rate and running speed were recorded during exercise. Venous blood samples were taken after a 10 h fast, at rest, after 10, 50 and 90 min of exercise as well as 10 and 30 min post-exercise. PRL, insulin, glucose, ammonia, lactate, triglycerides (TG), free fatty acids (FFA) and amino acids were determined in each sample. No significant differences were found in RPE. PRL increased (p < 0.01) in all TU. TG and heparin administration resulted in an increase (p <0.01) in FFA, which correlated (p < 0.01) with free TRP and the ratio of free TRP/TRP. Artificial increase in free TRP in TU III did not affect plasma PRL level. The amino acid infusion in TU II induced an increase in LNAA but had no significant effect on PRL. PRL and ammonia peaked at the end of exercise. We conclude that neither exercise-induced PRL secretion nor RPE are affected by changes in circulating free TRP and LNAA under the present conditions.

Adult↗
PubMed1996

Lymphocyte function and cytokine production during incremental exercise in active and sedentary males and females.

This study examined the effects of acute continuous incremental exercise on lymphocyte mitogenic function and cytokine production in physically active and sedentary males and females. Physically active (n = 32) and sedentary (N = 32) male and female subjects were randomly assigned to an exercise or control condition. Exercise involved a continuous incremental protocol consisting of cycling for 3 periods of 6 min at workrates corresponding to 55%, 70% and 85% VO2peak. Blood samples were drawn from a venous catheter at baseline, 6 min, 12 min and 18 min, and 2 h following completion of exercise. Relative to baseline and control condition the percentage of T (CD3+) and B cells (CD19+) significantly decreased, and the percentage of NK cells (CD3-CD16+CD56+) increased (p < 0.001) during each stage of the incremental exercise test. The proliferative response to ConA was suppressed, enhanced, or unchanged using 1.25 micrograms/ml, 2.5 micrograms/ml and 5.0 micrograms/ml ConA, respectively. The in-vitro production of IL-1 and IFN-gamma increased during each workload. In contrast IL-4 production did not change during exercise. The resting and exercise induced alterations in lymphocyte function and cytokine production were independent of gender and fitness level, and returned to baseline 2 h into recovery. The in-vitro production of IFN-gamma and IL-4 suggests that physical activity may alter the balance of TH1 and TH2 lymphocytes.

Adult↗
PubMed1996

The structure of the sarcomeric M band: localization of defined domains of myomesin, M-protein, and the 250-kD carboxy-terminal region of titin by immunoelectron microscopy.

The M band of vertebrate cross-striated myofibrils has remained an enigmatic structure. In addition to myosin thick filaments, two major structural proteins, myomesin and M-protein, have been localized to the M band. Also, titin is expected to be anchored in this structure. To begin to understand the molecular layout of these three proteins, a panel of 16 polyclonal and monoclonal antibodies directed against unique epitopes of defined sequence was assembled, and immunoelectron microscopy was used to locate the position of the epitopes at the sarcomere level. The results allow the localization and orientation of defined domains of titin, myomesin, and M-protein at high resolution. The 250-kD carboxy-terminal region of titin clearly enters the M band with the kinase domain situated approximately 52 nm from the central M1-line. The positions of three additional epitopes are compatible with the view that the titin molecule reaches approximately 60 nm into the opposite sarcomere half. Myomesin also seems to bridge the central M1-line and is oriented parallel to the long axis of the myofibril. The neighboring molecules are oriented in an antiparallel and staggered fashion. The amino-terminal portion of the protein, known to contain a myosin binding site, seems to adopt a specific three-dimensional arrangement. While myomesin is present in both slow and fast fibers, M-protein is restricted to fast fibers. It appears to be organized in a fundamentally different manner: the central portion of the polypeptide is around the M1-line, while the terminal epitopes seem to be arranged along thick filaments. This orientation fits the conspicuously stronger M1-lines in fast twitch fibers. Obvious implications of this model are discussed.

Animals↗
PubMed1996

The association of annexin I with early endosomes is regulated by Ca2+ and requires an intact N-terminal domain.

Annexin I is a member of a multigene family of Ca2+/phospholipid-binding proteins and a major substrate for the epidermal growth factor (EGF) receptor kinase, which has been implicated in membrane-related events along the endocytotic pathway, in particular in the sorting of internalized EGF receptors occurring in the multivesicular body. We analyzed in detail the intracellular distribution of this annexin by cell fractionation and immunoelectron microscopy. These studies used polyclonal as well as a set of species-specific monoclonal antibodies, whose epitopes were mapped to the lateral surface of the molecule next to a region thought to be involved in vesicle aggregation. Unexpectedly, the majority of annexin I was identified on early and not on multivesicular endosomes in a form that required micromolar levels of Ca2+ for the association. The specific cofractionation with early endosomes was also observed in transfected baby hamster kidney cells when the intracellular fate of ectopically expressed porcine annexin I was analyzed by using the species-specific monoclonal antibodies in Western blots of subcellular fractions. Interestingly, a truncation of the N-terminal 26, but not the N-terminal 13 residues of annexin I altered its intracellular distribution, shifting it from fractions containing early to those containing late and multivesicular endosomes. These findings underscore the regulatory importance of the N-terminal domain and provide evidence for an involvement of annexin I in early endocytotic processes.

Amino Acid Sequence↗
PubMed1996

Systemic sclerosis in German uranium miners under special consideration of autoantibody subsets and HLA class II alleles.

Systemic sclerosis (scleroderma) is a connective tissue disease with a wide range of clinical manifestations, with high or low degrees of skin and internal organ involvement together with different antinuclear antibody (ANA) specificities. Several studies provide evidence that males, who are rarely affected by systemic sclerosis, have an increased risk when working in mines. Therefore we reinvestigated 21 male subjects and 6 cases of deceased male patients who had been engaged in East German uranium mines and had shown evidence of this disease in medical examinations. Dermatological investigations, evaluation of chest X-rays and autoantibody estimation were performed. PCR-sequence-specific oligonucleotide typing was used to study the genetic association of HLA-D alleles with autoantibodies typical for scleroderma in these uranium miners suffering from systemic sclerosis and in patients with idiopathic systemic sclerosis. The determined HLA phenotype frequencies and the following statistical analysis (Fisher's exact test (2-sided)) revealed that in comparison with randomly selected controls, alleles DRB1*0300 (DR3) and DQB1*0201 (DQ2) were distinctly increased in the group of affected uranium miners, especially in those with anti-Scl-70 positivity. In contrast, we did not observe significant differences between affected and unaffected miners. Comparing anti-Scl-70-positive affected uranium miners with anti-Scl-70-positive idiopathic systemic sclerosis cases. DRB1*0300 as well as DQB1*0201 were also significantly enhanced in the former group. ACA-positive systemic sclerosis miners had significantly elevated frequencies in DRB1*0100 (DR1) and DRB1*0800 (DR8) only in comparison with unaffected miners and unexposed controls. Our genetic and immunological data lead to the assumption that a different set of HLA-D alleles in combination with exogenous factors is involved in the induction of anti-Scl-70 antibodies in uranium miners that might influence their susceptibility to the disease, whereas the same occupational exposure seems to have no influence on the induction of ACA antibodies.

Adult↗
PubMed1996

The central Z-disk region of titin is assembled from a novel repeat in variable copy numbers.

The giant sarcomeric protein titin (also described as connectin) is composed mainly of immunoglobulin (Ig)-like and fibronectin type III (fn3)-like domains arranged consecutively. At both ends of the molecule, these domains are interrupted by sequence insertions. The amino terminus of titin is localized in the Z-disk, a structure of great variability in different muscle types. We have determined the ultrastructural position of sequences in this region of the molecule in skeletal and cardiac muscle by immunoelectron microscopy using antibodies directed against unique epitopes. Titin molecules entering the Z-disk from two half sarcomeres do not significantly overlap, showing that the amino terminus is at the centre of the Z-disk. A serine/proline rich site, which can be phosphorylated by kinases in developing muscle tissues, was identified near the amino terminus of titin. Sequence analysis revealed the presence of a novel 45 residue repeat ('Z-repeats') in this region of the molecule. The number of titin Z-repeats varies due to differential splicing. We propose that this mechanism is a means of assembling Z-disks of variable thickness and mechanical strength.

Amino Acid Sequence↗
PubMed1996

The cruciated microtubule-associated fibers of the green alga Dunaliella bioculata consist of a 31 kDa SF-assemblin.

Cytoskeletons of Dunaliella bioculata, the biflagellate wallless green alga, were isolated and analyzed using a monoclonal and a polyclonal antibody raised against SF-assemblin, the major protein of the two striated microtubule-associated fibers of the alga Spermatozopsis similis. Indirect immunofluorescence showed antigenic structures associated with the four microtubular flagellar roots. SDS-PAGE followed by immunoblot analysis revealed a cross-reacting polypeptide of 31 kDa. This protein of D. bioculata was isolated using gel filtration chromatography in 8 M urea and in vitro reassembly of striated fibers. Microsequencing of the purified protein yielded various peptides, which could be aligned along the sequence of SF-assemblin from S. similis. A complete sequence of the Dunaliella protein was obtained by cDNA cloning. It documents the non helical head domain followed by a helical rod domain with a 29 residue repeat pattern based on four heptads followed by a skip residue. Compared to SF-assemblin of S. similis the SF-assemblin of Dunaliella has a shorter head and a slightly longer rod domain. The two algal SF-assemblins share only 57% sequence identity. We conclude that SF-assemblin and related proteins in various protists are representatives of a new class of alpha-helical proteins characterized by the ability to form a special segmented coiled coil and to assemble into striated fibers of 2 nm protofilaments in vivo and in vitro.

Amino Acid Sequence↗
PubMed1996

Monitoring gene therapy with cytosine deaminase: in vitro studies using tritiated-5-fluorocytosine.

UNLABELLED: Genetically modified mammalian cells that express the cytosine deaminase (CD) gene are able to convert the nontoxic prodrug 5-fluorocytosine (5-FC) to the toxic metabolite 5-fluorouracil (5-FU). PET with 18F-5-FC may be used for in vivo measurement of CD activity in genetically modified tumors. METHODS: A human glioblastoma cell line was stably transfected with the Escherichia coli CD gene. After incubation of lysates of CD-expressing cells and control cells with 3H-5-FC high-performance liquid chromatography (HPLC) was performed. The uptake of 5-FC was measured after various incubation times using therapeutic amounts of 5-FC. In addition, saturation and competition experiments with 5-FC and 5-FU were performed. Finally, the efflux was measured. RESULTS: We found that 3H-5-FU was produced in CD-expressing cells, whereas in the control cells only 3H-5-FC was detected. Moreover, significant amounts of 5-FU were found in the medium of cultured cells, which may account for the bystander effect observed in previous experiments. However, uptake studies revealed a moderate and nonsaturable accumulation of radioactivity in the tumor cells, suggesting that 5-FC enters the cells only through diffusion. Although a significant difference in 5-FC uptake was seen between CD-positive and control cells after 48 hr of incubation, no difference was observed after 2 hr of incubation. Furthermore, a rapid efflux could be demonstrated. CONCLUSION: 5-Fluorocytosine transport may be a limiting factor for this therapeutic procedure. Quantitation with PET has to rely more on dynamic studies and modeling, including HPLC analysis of the plasma, than on nonmodeling approaches.

Chromatography, High Pressure Liquid↗