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Biomedical subjects

C Hübner

Publications and source records attributed to C Hübner.

At least 55 records · Page 3Linked to original sources
PubMed1996

A cysteine 3340 substitution in the dystroglycan-binding domain of dystrophin associated with Duchenne muscular dystrophy, mental retardation and absence of the ERG b-wave.

We report the first C-terminal missense mutation in a Duchenne muscular dystrophy patient. A G10227A transition of the dystrophin gene was found which resulted in the substitution of a highly conserved cysteine at position 3340 within the second half of the dystroglycan-binding domain. Residual amounts of 427 kDa dystrophin were detected in western blot analysis of the patient's muscle tissue, and immunohistological examination revealed weak traces of dystrophin on all fibers. Sarcolemmal staining intensity of 43 kDa beta-dystroglycan was also reduced. Mental retardation in our patient and absence of the b-wave in his electroretinogram indicate that central nervous functions of dystrophin isoforms also depend on the presence of cysteine 3340.

Binding Sites↗
PubMed1996

Wheat kernel ingestion protects from progression of muscle weakness in mdx mice, an animal model of Duchenne muscular dystrophy.

A simple, reproducible test was used to quantify muscle weakness in mdx mice, an animal model of Duchenne muscular dystrophy. The effect of bedding on wheat kernels and of dietary supplementation of alpha-tocopherol on the progression of muscle weakness was investigated in mdx mice. When measured during the first 200 d of life, mdx mice developed muscle weakness, irrespective of bedding and diet. When kept on wood shavings and fed a conventional rodent diet, mdx mice showed progressive muscle weakness over the consecutive 200 d, and eventually showed a significant weight loss during the next 200-d observation period. Progression of muscle weakness and weight loss were almost completely prevented in mdx mice that were kept on wheat kernel bedding. In contrast, only incomplete maintenance of muscle strength and body weight was observed in mdx mice kept on wood shavings and fed the alpha-tocopherol-supplemented diet. It is concluded from these experiments that a component of wheat kernels other than alpha-tocopherol is essential to prevent the progression of muscle weakness in mdx mice.

Aging↗
PubMed1996

How different constituents of low density lipoprotein determine its oxidizability by copper: a correlational approach.

Although low density lipoprotein (LDL) susceptibility to oxidation is expected to be primarily related to its composition, the individual contributions of different constituents to its oxidizability remain unclear. The present study was undertaken to elucidate how different constituents of isolated LDL determine its susceptibility to oxidation induced by Cu2+ under conditions close to those of well-known Cu2(+)-oxidation assay (H. Esterbauer, G. Striegl, H. Puhl and M. Rotheneder (1989) Free Radical Research Communications, 6, 67-75). We characterized antioxidant, fatty acid and total lipid composition of human LDL from healthy donors (n = 22) and compared each with LDL oxidizability by Cu2+. LDL oxidizability was evaluated as oxidizability of antioxidant-containing LDL (rate of lipid peroxidation measured before total consumption of alpha-tocopherol, the major LDL antioxidant), oxidizability of antioxidant-depleted LDL (maximal rate of lipid peroxidation and maximal production of conjugated dienes within the propagation, antioxidant-depleted phase of oxidation) and overall LDL resistance to oxidation (duration of the lag-phase before the onset of the propagation phase). We found that the oxidizability of antioxidant-containing LDL correlated negatively with LDL content of ubiquinol-10 and free cholesterol, and positively with that of n-3 polyunsaturated fatty acids (PUFAs). LDL n-3 PUFAs, ubiquinol-10 and free cholesterol were the most important determinants of the oxidizability of antioxidant-containing LDL, contributing to about 35%, 25% and 25% of its total variability, respectively. Oxidizability of antioxidant-depleted LDL was largely determined by LDL PUFA content. The overall LDL resistance to oxidation correlated weakly with LDL chemical composition. alpha-Tocopherol was found to be only a minor contributor to the oxidizability of isolated LDL under oxidative conditions used (7.5 or 14 mol Cu2+ / mol LDL). It appears that the oxidizability of antioxidant-containing LDL represents a parameter highly sensitive to changing LDL composition, whereas the overall LDL resistance to oxidation combines contributions from different LDL constituents more uniformly, being weaker sensitive to individual factors. It is suggested that PUFAs, ubiquinol-10 and free cholesterol are the most important determinants of LDL oxidizability of Cu2+.

Adult↗
PubMed1996

Absorptive and secretory mechanisms in biliary epithelial cells.

Biliary epithelial cells line the intra- and extrahepatic biliary tree. They are involved in bile formation and are of importance in different cholestatic conditions (vanishing bile duct syndrome). In recent years, progress has been made to elucidate more precisely the physiological role of biliary epithelial cells. Biliary epithelial cells are involved in the cholehepatic shunting of bile acids, they reabsorb sugar and glutamate from bile. Secretin-induced electrolyte secretion is regulated by intracellular cAMP levels in these cells. Intracellular Ca2+ levels, which are increased by different agonists, e.g. carbachol, regulate transmembrane ion channels. ATP acts as secretagogue in these cells by activating ion channels and Na+, H+ antiporter. Further studies have to investigate in which parts of the biliary tree absorptive or secretory mechanisms are predominant.

Absorption↗
PubMed1995

Monitoring of ubiquinol-10, ubiquinone-10, carotenoids, and tocopherols in neonatal plasma microsamples using high-performance liquid chromatography with coulometric electrochemical detection.

A method for measuring the lipophilic antioxidants alpha-tocopherol, gamma-tocopherol, ubiquinol-10, ubiquinone-10, beta-cryptoxanthin, and beta-carotene in 5 or 10 microliters neonatal plasma is presented. First lipid extraction together with the internal standards gamma-tocotrienol, ubiquinone-7, ubiquinol-9, ubiquinone-9, and ethyl beta-apo-8'-carotenoate was carried out. Then the components were separated on a RPC2/C18 column. Coulometric electrochemical detection was performed using the reduction-oxidation mode (-0.60 V; -0.15 V; +0.60 V). Detection limits for all components were between 21 and 60 fmol, corresponding to 0.004 and 0.012 mumol/liter plasma; the within-day precision (coefficient of variation) was between 3 and 14% for all measured substances. Reference values are given for healthy newborn children from the second to the sixth day of life. Using the presented HPLC technique it is possible to monitor the concentrations of lipophilic antioxidants in plasma of sick immature newborn children in order to study effects of oxidative stress.

Antioxidants↗
PubMed1995

Antioxidative activity of ubiquinol-10 at physiologic concentrations in human low density lipoprotein.

Ubiquinol-10 is a powerful lipid-soluble antioxidant found in cell membranes and lipoproteins in vivo. Its mechanism of action on lipid peroxidation has been determined in model and biological systems. Data concerning antioxidative activity of ubiquinol-10 in lipoproteins, however, are still controversial. The present work examines its role in the prevention of low density lipoprotein (LDL) oxidation, specifically its influence on a copper-mediated oxidative modification of human LDL in vitro. We found that ubiquinol-10 incorporated in LDL in subnormal concentrations (0.05-0.13 mol/mol LDL incorporated in comparison with 0.10-1.20 mol/mol LDL reported as normally in human LDL) slightly but not significantly decreased production of lipid peroxidation products (lipid peroxides, conjugated dienes, thiobarbituric acid-reactive substances) during the first hours of oxidation. The extent of apolipoprotein B modification (LDL fluorescence at 360/430 nm) was also decreased. Increasing the ubiquinol-10 concentration in LDL to 0.55-1.48 mol/mol LDL made it significantly more resistant to copper-mediated oxidation than native LDL. Adding the same amounts of either ubiquinone-10 or alpha-tocopherol to the LDL suspension had almost no effect on its oxidation. Ubiquinol-10 decreased alpha-tocopherol consumption during LDL oxidation and was consumed more rapidly than the latter. These results demonstrate that LDL ubiquinol-10 content is an important factor influencing LDL susceptibility to oxidation by copper and suggest that it represents the first line of defense against oxidative modification in human LDL.

Antioxidants↗
PubMed1995

X-chromosome methylation in manifesting and healthy carriers of dystrophinopathies: concordance of activation ratios among first degree female relatives and skewed inactivation as cause of the affected phenotypes.

The X-chromosome activity states of 11 manifesting carriers of dystrophinopathies, all with normal karyotypes, were estimated by restriction fragment length polymorphism (RFLP)-methylation analysis with the probes M27 beta (DXS255), p2-19(DXS605) and pSPT/PGK (PGK1) to test the role of skewed X-inactivation ratios as the cause of their affected phenotypes. In eight cases preferential inactivation of the putative X chromosome carrying the normal dystrophin allele in > or = 90% of their peripheral lymphocytes was observed, two cases showed non-apparent deviant ratios (60:40 and 70:30) from the theoretically expected values around the mean of 50% and in one case the three markers employed yielded no information. The analysis of the X-inactivation ratio in six mother-daughter pairs, all non-manifesting Duchenne muscular dystrophy (DMD) carriers, and in the close female relatives of the patients showed: (a) neither of the two X chromosomes was preferentially inactivated with respect to their parental origin; (b) a high concordance among the activation ratios of mothers and daughters, a result difficult to explain just in terms of random X-chromosome inactivation.

Child↗
PubMed1995

In vitro effects of oxidized low density lipoprotein on CD11b/CD18 and L-selectin presentation on neutrophils and monocytes with relevance for the in vivo situation.

Oxidized LDL (oxLDL) has been identified as a potent stimulus of leukocyte adhesion to endothelium, a hallmark of early atherogenesis. A cytofluorometric study was performed to further characterize the mechanisms by which oxLDL stimulates the rapid adhesion of leukocytes to endothelium in vitro and in vivo. Incubation (30 minutes at 37 C) of whole blood (diluted with buffered saline to 1 x 10(6) leukocytes/ml) with oxLDL (0.85 mg LDL cholesterol/ml; oxidized by 7.5 mumol/L Cu2+ for 18 hours) but not native LDL stimulated the upregulation of CD11b/CD18 adhesion receptors on neutrophils (anti-leu-15 binding: 178 +/- 16% of baseline, P < 0.01, means +/- SD of n = 10 experiments) and on monocytes (169 +/- 34% of baseline, P < 0.01). This phenomenon was almost entirely inhibited by n-butanol or the vasoactive drug pentoxifylline (PTX), which also significantly reduced oxLDL-induced leukocyte adhesion to venular and arteriolar endothelium, as assessed by intravital microscopy on the dorsal skinfold chamber in hamsters (venules: 49 +/- 19 versus 120 +/- 34 cells/mm2, P < 0.05; arterioles: 9 +/- 4 versus 52 +/- 7 cells/mm2, P < 0.01) 30 minutes after intravenous injection of oxLDL (4 mg/kg body weight; means +/- SD of n = 7 hamsters per group). Butanol and PTX also significantly reduced the upregulation of CD11b/CD18 by f-methionyl-leucyl-phenylalanine (fMLP) and platelet-activating factor (PAF) but not by phorbol myristate acetate (PMA). Whereas fMLP and PAF stimulate leukocytes via binding to specific cell surface receptors and triggering complex signal transduction pathways, PMA bypasses these pathways and directly activates intracellular protein kinase C. By analogy, we propose that oxLDL upregulates CD11b/CD18 through its previously documented ability to stimulate the generation of second messengers. The effect of n-butanol and PTX on receptor presentation cannot be explained by changes in plasma membrane fluidity, as both agents failed to reverse the decrease in plasma membrane fluidity of neutrophils after stimulation with oxLDL, as assessed by fluorescence anisotropy measurement of the membrane marker diphenylhexatriene. Incubation of isolated neutrophils but not of whole blood with oxLDL resulted in a significant loss of L-selectin from the neutrophil surface (anti-TQ-1 binding: 40 +/- 13% of baseline, P < 0.01). A significant loss of this adhesion receptor on neutrophils and monocytes was also observed after stimulation of isolated neutrophils and whole blood with fMLP, PAF, and PMA.(ABSTRACT TRUNCATED AT 400 WORDS)

1-Butanol↗
PubMed1995

Deteriorating free radical-trapping capacity and antioxidant status in plasma during bone marrow transplantation.

Organ toxicity in BMT may in part be due to free radical damage. Therefore the 'Total Radical-trapping Antioxidant Parameter of plasma' (TRAP), individual plasma antioxidants, serum iron and linoleic acid, a main substrate of lipid peroxidation, were monitored before and after BMT, and they were compared with values obtained from healthy controls. Seven patients (3 AML, 3 CML, 1 multiple myeloma) receiving 16 mg/kg busulfan, 30-45 mg VP-16 and 120 mg/kg cyclophosphamide were investigated. TRAP values declined during chemotherapy by about 40% (day -9: 1019 +/- 245 mumol/l, mean +/- s.d.; day 0: 660 +/- 164 mumol/l; P < 0.05). The concentration of uric acid, one of the main antioxidants in plasma, decreased markedly (day -9: 339 +/- 108 mumol/l, day 0: 148 +/- 61 mumol/l; P < 0.05) and paralleled TRAP values. Vitamin E and bilirubin did not change from day -9 to 0 whereas vitamin C increased (day -9: 46 +/- 16 mumol/l, day 0: 89 +/- 44 mumol/l; P < 0.05). Serum iron rapidly increased within the pre-transplantation period, reaching values normally seen only in iron overload (day -9: 11.8 +/- 5.2 mumol/l, day 0: 40.6 +/- 6.5 mumol/l; P < 0.05). Linoleic acid levels were normal at the start and decreased substantially (27.0 +/- 1.6 wt% at day -9; 15.7 +/- 4.9 wt% at day 0; P < 0.05), indicating possible lipid peroxidation during high-dose chemotherapy. In conclusion, complex monitoring of the antioxidant status before and after BMT revealed a breakdown of plasma antioxidant defence and of radical-vulnerable lipids, which was associated with high circulating levels of iron.

Adult↗
PubMed1994

Low density lipoprotein oxidizability by copper correlates to its initial ubiquinol-10 and polyunsaturated fatty acid content.

At an early stage of oxidation induced by Cu2+, the rate of oxidative modification of human low density lipoprotein (LDL) from healthy donors correlated negatively to its ubiquinol-10 (r = -0.58, P < 0.01) and positively to its polyunsaturated fatty acid (PUFA) (r = 0.53, P < 0.05) content. The PUFA/ubiquinol-10 ratio was the best predictor of LDL susceptibility to oxidation (r = 0.68, P < 0.01). No significant correlation between LDL oxidizability and its alpha-tocopherol content was found at any oxidation stage. It is suggested that ubiquinol-10 plays a central role in the early protection of LDL PUFAs against Cu(2+)-induced oxidation whereas alpha-tocopherol posesses both pro- and antioxidant activity.

Adult↗
PubMed1994

A new experimental approach for monitoring postoperative bypass patency after coronary artery bypass grafting.

Little information is known concerning hemodynamic parameters of bypass grafts after open heart surgery during the postoperative period. As there is no easy to use bedside-method for monitoring, we developed a new implantable Doppler probe to measure blood flow velocity with a 20 MHz pulsed Doppler system. The accuracy of this method was proved in a circulatory model and in animal studies. Our conclusion is that the implantable pulsed Doppler system is a sensitive method for monitoring small vessel hemodynamics.

Animals↗
PubMed1994

The effect of ephedrine bolus administration on left ventricular loading and systolic performance during high thoracic epidural anesthesia combined with general anesthesia.

We investigated the effect of ephedrine on left ventricular function in patients without cardiovascular disease under high thoracic epidural anesthesia combined with general anesthesia. Because the epidural block was extended to all cardiac segments, ephedrine was assumed to be deprived of its centrally mediated actions. Left ventricular (LV) function was assessed using transesophageal echocardiography. We measured arterial pressure (AP), heart rate (HR), LV end-systolic and end-diastolic diameter and area (ESA, EDA), wall thickness, and LV ejection time before and after intravenous ephedrine bolus administration. We calculated area ejection fraction (EFA), end-systolic wall stress (ESWS), and mean velocity of circumferential fiber shortening (mVcfc). Ephedrine had a biphasic effect on left ventricular function. It transiently decreased EDA from 18.9 to 16.5 cm2 (mean), whereas EFA and mVcfc were increased from 33% to 49%, and from 1.88 to 2.67 circumferences/s, respectively. During the second phase, ephedrine increased mean arterial pressure (MAP) from a baseline value of 62 to 87 mm Hg, EDA was restored to 19.3 cm2, and EFA and mVcfc remained above baseline (52% and 2.64 circumferences/s, respectively). ESWS was not significantly increased from baseline. We conclude that ephedrine improves left ventricular contractility, even in the presence of high thoracic epidural anesthesia, without causing relevant changes of left ventricular afterload.

Adult↗
PubMed1994

P-selectin mediates the interaction of circulating leukocytes with platelets and microvascular endothelium in response to oxidized lipoprotein in vivo.

BACKGROUND: Oxidized low density lipoprotein (oxLDL) has been demonstrated to stimulate leukocyte/endothelium interaction, an early feature of atherogenesis. Using the skinfold chamber model for intravital microscopy in hamsters and mice, we have shown that oxLDL-induced leukocyte adhesion to microvascular endothelium shares many characteristics with leukocyte adhesion during inflammation and ischemia/reperfusion, including the involvement of beta 2 integrin adhesion molecules. In light of the two-step model of leukocyte adhesion, we have examined the contribution of P-selectin to oxLDL-induced leukocyte/endothelium interaction. P-selectin is an inducible adhesion molecule on platelets and endothelium, mediating the initial steps of leukocyte margination and rolling along the endothelial lining, as well as of aggregate formation between platelets and leukocytes. EXPERIMENTAL DESIGN: For our studies, we used the dorsal skinfold chamber model for intravital fluorescence microscopy on awake Syrian golden hamsters. Hamsters were treated 10 minutes before oxLDL-injection (oxidized by Cu2+, 4 mg/kg body weight, intravenously) with blocking antibodies to P-selectin (2 mg/kg body weight intravenously, N = 7). RESULTS: In seven control animals (pretreated with an irrelevant IgG antibody), oxLDL injection elicited leukocyte rolling and adhesion on both venular and arteriolar endothelium, and also the formation of aggregates tumbling down the microvessels and firmly adhering to the microvascular endothelium. The aggregates consisted of leukocytes and activated, dendritic platelets, as assessed by scanning electron microscopy of the buffy coat isolated by density gradient centrifugation of whole blood taken from hamsters 15 minutes after injection of oxLDL. Leukocyte adhesion to venular and arteriolar endothelium, as well as the formation of leukocyte/platelet aggregates were significantly reduced by pretreatment of the animals with anti-P-selectin antibodies. CONCLUSIONS: These data emphasize the similarities between leukocyte adhesion in response to oxLDL and in other pathophysiologic conditions, identifying P-selectin as a crucial player in the interaction between leukocytes and microvascular endothelium as well as in the formation of circulating leukocyte/platelet aggregates.

Animals↗
PubMed1994

Effects of vasoactive drugs on postoperative coronary bypass flow: a new technique.

OBJECTIVE: This study aimed to investigate the influence of vasoactive drugs, including a calcium channel blocker, nitroglycerine, and a beta-blocker, often used after coronary artery bypass grafting (CABG). Measurements were taken using a new, completely implantable, mini-Doppler system. The probes were implanted during CABG, led outside through the thoracic wall, and removed 3 days after the operation by a simple pull. RESULTS: The mean Doppler flow increased dramatically in the calcium channel blocker and the nitroglycerine groups (P = 0.002) and decreased slightly in the beta-blocker group (P = 0.015). Mean arterial pressure decreased significantly in all three groups. Heart rate decreased in the beta-blocker group (P = 0.002). Changes in the other hemodynamic variables measured were not significant; there were no changes in the control group. Doppler sonographic monitoring of bypass diameters did not show any changes. CONCLUSION: This is the first study to measure the influence of vasoactive drugs directly and online after CABG. From our experience of more than 40 implantations, we conclude that our method is easy to use and reliable and will help improve the therapeutic regimen for patients after CABG.

Aged↗
PubMed1993

Multipoint mapping of the central core disease locus.

A linkage analysis with 12 DNA markers from proximal 19q was performed in eight families with central core disease (CCO). Two-point analysis gave a peak lod score of Z = 4.95 at theta = 0.00 for the anonymous marker D19S190 and of Z = 2.53 at theta = 0.00 for the ryanodine receptor (RYR1) candidate gene. Multipoint linkage data place the CCO locus at 19q13.1, flanked proximally by D19S191/D19S28 and distally by D19S47. This map location includes the RYR1 gene. The results of the linkage study present no evidence for genetic heterogeneity of CCO.

Animals↗
PubMed1993

Cigarette smoke elicits leukocyte adhesion to endothelium in hamsters: inhibition by CuZn-SOD.

Although cigarette smoking has been identified as a major risk factor for cardiovascular diseases, the underlying pathomechanism is largely unknown. Using a dorsal skinfold chamber model in Syrian golden hamsters for intravital microscopy on striated muscle microcirculation, we investigated whether cigarette smoke (CS) affects the adhesion of circulating leukocytes to the endothelium, a constant feature of early atherogenesis and a hallmark of ischemia-reperfusion injury. Awake hamsters were exposed for 5 min to the mainstream smoke of one cigarette (2R1 research cigarette), inducing nicotine, cotinine, and carboxyhemoglobin plasma levels comparable to levels found in human smokers. In control animals (n = 7), CS exposure elicited the rolling and subsequent adhesion of fluorescently stained leukocytes to the endothelium of arterioles and postcapillary venules. Leukocyte/endothelium interaction was preceded by an early rise in xanthine oxidase activity and intravascular hemolysis. Leukocyte adhesion and xanthine oxidase (XO) activation were significantly attenuated in hamsters pretreated with superoxide dismutase (5 mg/kg, 10 min prior to CS, n = 7), suggesting a key role of superoxide in this event. These in vivo results suggest a novel pathomechanism of CS-induced cardiovascular pathology.

Animals↗