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C Bachmann

Publications and source records attributed to C Bachmann.

At least 37 records · Page 2Linked to original sources
PubMed1998

Analysis and regulation of vasodilator-stimulated phosphoprotein serine 239 phosphorylation in vitro and in intact cells using a phosphospecific monoclonal antibody.

The development and functional analysis of a monoclonal antibody (16C2) are reported; the antibody recognizes vasodilator-stimulated phosphoprotein (VASP; an established substrate of both cAMP- and cGMP-dependent protein kinase) only when serine 239 is phosphorylated. VASP serine 239 represents one of the best characterized cGMP-dependent protein kinase phosphorylation sites in vitro and in intact cells. Experiments with purified, recombinant human VASP and various VASP constructs with mutated phosphorylation sites (S157A, S239A, T278A) and experiments with intact cells (human/rat platelets and other cells) treated with cyclic nucleotide-elevating agents demonstrated the specificity of the monoclonal antibody 16C2. Quantitative analysis of the VASP shift from 46 to 50 kDa (indicating VASP serine 157 phosphorylation) and the appearance of VASP detected by the 16C2 monoclonal antibody (VASP serine 239 phosphorylation) in human platelets stimulated by selective protein kinase activators confirmed that serine 239 is the VASP phosphorylation site preferred by cGMP-dependent protein kinase in intact cells. Immunofluorescence experiments with human platelets treated with cGMP analogs showed that the 16C2 monoclonal antibody also detects VASP serine 239 phosphorylation in situ at established intracellular localization sites. Analysis of VASP serine 239 phosphorylation by the 16C2 antibody appears to be the best method presently available to measure cGMP-dependent protein kinase activation in intact cells. Also, the 16C2 antibody promises to be an excellent tool for the evaluation of VASP function in intact cells.

Animals↗
PubMed1998

Clinical chemistry variables in normal elderly and healthy ambulatory populations: comparison with reference values.

Laboratory values of the most commonly assayed clinical chemistry variables were determined in selected elderly and healthy ambulatory populations. The upper and lower limits (2.5 and 97.5 fractiles) were compared with the adult reference values in use in university hospitals of Switzerland. The results suggest that conventional adult reference values can be used for most variables in the elderly and that these values are also useful in an ambulatory population.

Aged↗
PubMed1998

Effects of a partially hydrolyzed guar gum on intestinal absorption of carbohydrate, protein and fat: a double-blind controlled study in volunteers.

UNLABELLED: Some dietary fibres have been shown to affect the rate of absorption of dietary carbohydrate, protein and fat as well as exocrine pancreatic function. The aim of the study was to examine the effect of partially hydrolysed guar gum (BENEFIBER), on normal absorption of glucose, amino acid (arginine) and fat. In addition hepatic, pancreatic, renal and haematological side effects were evaluated. METHODS: A double blind, randomized and cross-over design was used. Each subject served as its own control. Ten healthy male volunteers participated in the study. Each subject was randomly assigned to two different dietary regimes for a period of 7 days each. The study was repeated with the other diet for another 7-day period after an interval of at least 1 week. In one study period the subjects took liquid formula diet without fibre and during the other study period they took the same diet with fibre. RESULTS: The results of the study demonstrated that BENEFIBER did apparently not interfere with the normal absorption of glucose, amino acid and fat. In keeping with these observations, insulin release and exocrine pancreatic function were not affected. Haematological, renal and hepatic toxicity were not observed in association with BENEFIBER consumption. CONCLUSION: We conclude that BENEFIBER is a safe source of soluble fibre.

Adult↗
PubMed1998

Mutations in Drosophila enabled and rescue by human vasodilator-stimulated phosphoprotein (VASP) indicate important functional roles for Ena/VASP homology domain 1 (EVH1) and EVH2 domains.

Drosophila Enabled (Ena) was initially identified as a dominant genetic suppressor of mutations in the Abelson tyrosine kinase and, more recently, as a member of the Ena/human vasodilator-stimulated phosphoprotein (VASP) family of proteins. We have used genetic, biochemical, and cell biological approaches to demonstrate the functional relationship between Ena and human VASP. In addition, we have defined the roles of Ena domains identified as essential for its activity in vivo. We have demonstrated that VASP rescues the embryonic lethality associated with loss of Ena function in Drosophila and have shown that Ena, like VASP, is associated with actin filaments and focal adhesions when expressed in cultured cells. To define sequences that are central to Ena function, we have characterized the molecular lesions present in two lethal ena mutant alleles that affected the Ena/VASP homology domain 1 (EVH1) and EVH2. A missense mutation that resulted in an amino acid substitution in the EVH1 domain eliminated in vitro binding of Ena to the cytoskeletal protein zyxin, a previously reported binding partner of VASP. A nonsense mutation that resulted in a C-terminally truncated Ena protein lacking the EVH2 domain failed to form multimeric complexes and exhibited reduced binding to zyxin and the Abelson Src homology 3 domain. Our analysis demonstrates that Ena and VASP are functionally homologous and defines the conserved EVH1 and EVH2 domains as central to the physiological activity of Ena.

Amino Acid Sequence↗
PubMed1997

Microheterogeneity of serum glycoproteins and their liver precursors in patients with carbohydrate-deficient glycoprotein syndrome type I: apparent deficiencies in clusterin and serum amyloid P.

Serum and liver protein patterns were studied, respectively, in 5 patients (serum) and 1 patient (liver) with carbohydrate-deficient glycoprotein syndrome (CDGS) type I by high-resolution two-dimensional electrophoresis (2-DE) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The pattern of serum glycoproteins in all 5 patients presented abnormal trains of isoforms with decreased mass (delta molecular weight 3000) and all showed a cathodal shift. Two-dimensional electrophoresis and SDS-PAGE mass analysis of transferrin, alpha1 -antitrypsin, haptoglobin beta-chain, and alpha1-acid glycoprotein after neuraminidase and N-glycosidase F treatments demonstrated that the additional trains of the isoforms found in CDGS type I contain homologous species of isoforms. Some of them still showed charge differences, and all still contained glycans except for transferrin, with some unusual nonglycosylated isoforms. In addition, deficiencies in clusterin and serum amyloid P, not described so far, have been found in all 5 patients. The two-dimensional pattern of immunodetected precursors of serum proteins in liver cells from 1 patient with CDGS showed abnormal low-mass precursors and the absence of the precursors normally found in controls. These results suggest that these abnormal precursors accumulate during the early oligosaccharide processing of the nascent protein-bound oligosaccharides and that glycoprotein precursors undergo an altered intracellular transport while the post-translational processing along the normal pathway is still apparently functioning in patients with CDGS.

Amyloid↗
PubMed1997

Urinary phosphate/creatinine, calcium/creatinine, and magnesium/creatinine ratios in a healthy pediatric population.

OBJECTIVE: To determine reference values for urinary phosphate/creatinine (Cr) concentration ratios and to complete reference values for urinary calcium/creatinine and magnesium/creatinine ratios in the second morning urine sample of healthy infants, children, and adolescents. DESIGN: Urinary P/Cr, Ca/Cr, and Mg/Cr ratios were determined from the second morning urine sample. Two urine samples were obtained 1 week apart from most subjects to assess reproducibility. SETTING: Kindergartens and schools of Lausanne, Switzerland. PARTICIPANTS: A total of 410 healthy children aged 1 month to 17 years (197 girls and 213 boys) participated in the study. RESULTS: The 5th and 95th percentiles were estimated from 664 urine samples. There were no differences related to sex. A nonlinear regression in terms of age was used to smooth the estimated percentiles yielding reference curves from which critical values may be obtained for any given age. The 95th percentile for urinary Ca/Cr and Mg/Cr agreed with previously reported values in children older than 7 years. The upper limit of the three solute/creatinine ratios decreased significantly with age: for urinary P/Cr from 19.0 mol/mol at 1 month to 2.7 at 14 years; for urinary Ca/Cr from 2.2 to 0.7 mol/mol, and for urinary Mg/Cr from 2.2 to 0.6 mol/mol. Lower limits varied little. Interindividual and intraindividual variations decreased with age. CONCLUSIONS: Urinary P/Cr, Ca/Cr, and Mg/Cr ratios vary strongly with age. We provide reference values, expressed both in SI and in mass units, for urinary P/Cr, Ca/Cr, and Mg/Cr in children aged one month to 17 years.

Adolescent↗
PubMed1996

Tyrosine uptake and regional brain monoamine metabolites in a rat model resembling congenital hyperammonemia.

Hyperammonemia found in congenital disorders has a toxic effect on the central nervous system. Disturbances of brain neurotransmitter metabolism have been proposed, such as an increased transport of tryptophan into the brain and an increased flux through the serotonin pathway. Results concerning the catecholamine pathway are, however, contradictory. We therefore studied whether hyperammonermia increases brain uptake of the neurotransmitter precursor amino acid tyrosine and whether these changes affect the concentration of neurotransmitters and their metabolites in different brain areas (frontal cortex, caudatus-putamen, thalamus, hypothalamus, hippocampus/substantia nigra, brainstem) of rats made hyperammonemic with urease. The brain uptake of tyrosine was measured in the forebrain, brainstem, and cerebellum. The brain areas were analyzed for dopamine, 3,4-hydroxyphenylacetic acid; homovanillic acid, norepinephrine, and vanillylmandelic acid. The brain uptake index of tyrosine was increased in the forebrain and brainstem of the hyperammonemic rats with concomitantly elevated concentrations in the forebrain of tyrosine, phenylalanine, and tryptophan. The homovanillic acid content was significantly increased in the hypothalamus, hippocampus/substantia nigra and brainstem. The concentrations of norepinephrine, dopamine, and 3, 4-hydroxyphenylacetic acid were not significantly changed. Vanillylmandellic acid was decreased in the caudatus-putamen, thalamus, and hypothalamus. The data indicate an undisturbed neurotransmitter synthesis and, taken with the augmented tyrosine uptake at the blood-brain barrier, an increased flux through the dopamine pathway. These changes observed in the hyperammonemic animal model could contribute to the understanding of the pathogenic mechanisms and offer an explanation for the neuropsychiatric disturbances observed in children with congenital hyperammonemia.

Amino Acids↗
PubMed1995

Reduction of plasma alanine aminotransferase during vigabatrin treatment.

In 9 drug-resistant patients with partial seizures treated with vigabatrin, gamma-vinyl GABA (VGB), alanine aminotransaminase (ALAT) activity in plasma was significantly reduced. Comparison of in vitro with in vivo measurements led us to conclude that this reduction is mainly an in vivo phenomenon, perhaps due to cross-enzyme inhibition. The assessment of two biological variables linked with ALAT, glucose and alanine levels under fasting conditions, failed to show any significant metabolic alterations. VGB is an effective drug for partial epilepsy. Our observations do not suggest that reduced ALAT activity is of clinical concern.

Adolescent↗
PubMed1995

Interaction of fluorescein derivatives with sulfonylurea binding in insulin-secreting cells.

Recently evidence was presented that fluorescein derivatives (e.g. phloxine B) inhibit glibenclamide binding by occupation of a nucleotide-binding site at the ATP-sensitive potassium channel (KATP channel). However, this conclusion was inconsistent with the results of previous studies testing the effects of nucleotides on glibenclamide binding. To elucidate the interaction mode of fluorescein derivatives with sulfonylurea binding, the effect of phloxine B on binding of [3H]glibenclamide to microsomes obtained from a pancreatic beta-cell line (HIT-T15) was examined. Phloxine B inhibited specific binding of glibenclamide half-maximally at 3.2 mumol/l. The slope parameter for the displacement curve was close to one, suggesting a competitive interaction between both drugs. In accordance with this assumption 4 mumol/l phloxine B did not show an effect on the number of high-affinity binding sites but increased the apparent dissociation constant for glibenclamide by 3.1-fold and 30 mumol/l phloxine B did not alter the rate of dissociation of [3H]glibenclamide. Moreover, MgATP (300 mumol/l) significantly reduced the apparent affinity for binding of phloxine B to the sulfonylurea receptor. This finding resembled the action of MgATP on binding of sulfonylureas to their receptor site. It is concluded that fluorescein derivatives inhibit glibenclamide binding due to competition for the same site at the sulfonylurea receptor.

Benzamides↗
PubMed1994

The treatment of spondylotic cervical myelopathy by ventral discectomy. Long term results on 121 patients.

The pathophysiology of spondylotic cervical myelopathy is still a matter of discussion. This paper presents a series of 126 patients operated on using a ventral approach. In 47% of the patients only a spondylotic narrowing of the spinal canal was present and in 35% an additional disc herniation was found. In 13% of the cases however a soft disc without spondylotic spures was found and in 5% a dislocation of vertebral bodies. We found a marked male preponderance of 77%, mean age was 51.6 years, ranging from 25-50 years. Most patients were operated on at the levels of C4/5 and C5/6. Observation time covered a period of 3-10 years. The outcome was rated relatively to the preoperative degree of disablement using a questionnaire for the patients and their family doctors. We found a marked difference in the answers, especially in rating deterioration, which was stated by patients in 34%, by physicians only in 12%. Another finding was the time-related outcome. We found best results with 75% improvement and 5% deterioration between 3-6 months postoperatively, with increasing time the results decreased to 33% improvement, 33% identical statys and in 33% a deterioration related to the preoperative status must be noted.

Adult↗
PubMed1994

Pitfalls in aminoacid and organic acid analysis: 3-hydroxypropionic aciduria.

Pitfalls in organic acid analysis can originate from inadequate methodology, analytical interferences, in vivo interactions and from pre-analytical conditions which often are unknown to the specialized analytical laboratory. Among the latter, ingested food and additives, metabolites of food processing or medications have to be considered. Bacterial metabolites from the gastrointestinal or urogenital system or formed after sample collection can lead to pitfalls as well. An example of such a patient whose urinary metabolites mimic at first glance inherited propionic aciduria is described.

Amino Acid Metabolism, Inborn Errors↗
PubMed1994

Photoaffinity labeling of the cerebral sulfonylurea receptor using a novel radioiodinated azidoglibenclamide analogue.

In previous studies evidence has been presented by photoaffinity labeling that a polypeptide of 145-150 kDa represents the cerebral sulfonylurea receptor. However, covalent incorporation of [3H]glibenclamide or a 125I-labeled glibenclamide analogue into the sulfonylurea receptor required high amounts of photoenergy and took place with low yield of photoinsertion. To provide a probe with increased photoreactivity a 4-azido-5-iodosalicyloyl analogue of glibenclamide was synthesized. Binding experiments revealed specific and reversible high-affinity binding of this novel probe to the particulate (KD = 0.13 nM) and solubilized (KD = 0.56 nM) sulfonylurea receptor from cerebral cortex. The novel probe showed > 100-fold higher sensitivity to irradiation at 356 nm than glibenclamide. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed specific photoincorporation into a cerebral protein of 175 kDa and indicated an efficiency of photoincorporation of 9%. From dissociation binding curves following irradiation photoincorporation was estimated as 28% of specifically bound ligand. Photoincorporation into the 175-kDa protein following saturation binding of the novel probe to particulate sites from cerebral cortex indicated a KD value of 0.38 nM. Inhibition of photoincorporation into this protein by glibenclamide, glipizide, and tolbutamide revealed KD values for these sulfonylureas of 0.06 nM, 1.6 nM, and 1.2 microM, respectively. These results show that the novel photoaffinity ligand can be used as a probe for detection and characterization of the sulfonylurea receptor and suggest that a 175-kDa protein represents the cerebral sulfonylurea receptor.

ATP-Binding Cassette Transporters↗
PubMed1994

Age-related reference values for urinary organic acids in a healthy Turkish pediatric population.

Organic acid concentrations were quantified by gas chromatography and the individual acids identified by mass spectrometry in urine specimens from a healthy Turkish pediatric population of ages 2 days to 16 years, subdivided into five age groups. We quantified 69 organic acids (32 major and 37 minor components) and report here the median values and percentiles for each compound, adjusted for creatinine content, for the five groups. Concentrations of most of the organic acids tend to decrease with age but display substantial differences between age groups. This emphasizes the importance of comparing patients' data with age-matched reference data. Correlations between the excretion of organic acids and protein or caloric intake were significant for several compounds.

Aging↗