Biomedical subjects

G Schwarz

Publications and source records attributed to G Schwarz.

At least 19 recordsLinked to original sources

PubMed2005

[Bile acid-independent effect of hymecromone on bile secretion and common bile duct motility].

BACKGROUND AND OBJECTIVE: Hymecromone (4-methyl-umbiliferone) has been used for more than 20 years for the treatment of functional and obstructive spasms of the biliary tract. Its mode of action however is still largely unknown. We investigated the effect of 4-methyl-umbiliferone p. o. and i. v. on gall bladder and common bile duct motility and studied potentially indirect effects via alterations in bile acid metabolism. PATIENTS AND METHODS: Twenty healthy volunteers, aged 25 - 37, 10 males, 10 females, were included into a Placebo-controlled, randomised, cross-over double-blind study. Subjects were treated with 800 mg hymecromone p. o.; in addition a standardized meal (Biloptin, 40 gs) was given. Gall bladder volume and common bile duct diameter were determined by ultrasound. Conjugated and unconjugated bile acids were analysed by gas chromatography. Additionally, in a third open label phase hymecromone was given i. v. RESULTS: Common bile duct diameter was significantly larger after a standard meal with hymecromone given p. o. or i. v. than with placebo (each p < 0.01). However, alterations in gall bladder volume after a standard meal were not different between placebo and hymecromone (p. o. or i. v.). Unconjugated and conjugated bile acids rose after standard meal in all three groups without significant differences between hymecromone and placebo. CONCLUSIONS: Hymecromone was associated with significant dilation of the common bile duct. In contrast to previous reports an effect of hymecromone on gall bladder motility could not be observed. The unchanged values of bile acids in serum after hymecromone compared to placebo, together with the dilatation of the common bile duct after hymecromone, may indicate a bile acid-independent effect of hymecromone on bile secretion.

Administration, Oral↗

PubMed2005

Molybdenum cofactor biosynthesis and deficiency.

The molybdenum cofactor (Moco) forms the active site of all molybdenum (Mo) enzymes, except nitrogenase. Mo enzymes catalyze important redox reactions in global metabolic cycles. Moco consists of Mo covalently bound to one or two dithiolates attached to a unique tricyclic pterin moiety commonly referred to as molybdopterin (MPT). Moco is synthesized by an ancient and conserved biosynthetic pathway that can be divided into four steps, according to the biosynthetic intermediates precursor Z (cyclic pyranopterin monophosphate), MPT and adenylated MPT. In a fifth step modifications such as attachment of nucleotides, sulfuration or bond formation between Mo and the protein result in different catalytic Mo centers. A defect in any of the steps of Moco biosynthesis results in the pleiotropic loss of all Mo enzyme activities. Human Moco deficiency is a hereditary metabolic disorder characterized by severe neurodegeneration resulting in early childhood death. Recently, a first substitution therapy was established.

Animals↗

PubMed2005

[GCP inspections of clinical trials].

The new regulations on clinical trials with medicinal products for human use include rules for Good Clinical Practice (GCP) inspections. These rules pertain to such inspections as part of the surveillance program of the relevant state authorities, in preparation for a decision on the authorization of clinical trials by the federal authorities, and pre and post approval GCP-inspections as one tool of Good Regulatory Practice (GRP). The overall aim of such inspections is to verify compliance with the provisions on GCP and, thus, to ensure the protection of trial subjects, the credibility and reliability of clinical trial results, and the pro-active protection of patients. Since inspection results have to be acknowledged by other member states, there is an increasing importance for quality assurance and harmonization of inspection procedures. As in most cases a considerable number of sites are involved in the conduct of clinical trials in different member states, there is a need for coordination of GCP inspections between these states.

Clinical Protocols↗

PubMed2004

Development and validation of a PCR-based marker assay for negative selection of the HMW glutenin allele Glu-B1-1d (Bx-6) in wheat.

Polymorphisms between the coding sequences of high-molecular-weight (HMW) glutenin x-type genes at the Glu-1 locus were used to amplify Glu-1B x-type-specific PCR fragments. PCR analysis in a wheat cultivar subset carrying different Glu-1B x-type alleles resulted in PCR fragments that differed in size for Glu-B1-1d (B-x6) and non -Glu-B1-1d (B-x6) genotypes. Subsequent sequencing analysis revealed a 15-bp in-frame insertion in the coding regions of all Glu-B1-1d (B-x6) genotypes which allowed the development of a B-x6-specific PCR assay for high-throughput allele sizing by ion-pair reversed-phase high-performance liquid chromatography. The assay was validated in a set of 86 German wheat cultivars, and genotyping data unequivocally verified the presence of HMW glutenin subunits GLU-B1-1D (Bx-6) + GLU-B1-2A (By-8) by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis. These results demonstrate that the PCR assay can be applied for the detection and negative selection of the 'poor breadmaking quality' Glu-B1-1d (B-x6) alleles in wheat breeding programs.

Alleles↗

PubMed2004

[Quantification of gender specific thermal sensory and pain threshold before and after laser needle stimulation].

Quantitative thermal sensory and pain threshold testing (QST) was performed in 29 adult healthy volunteers (mean age 24.2 +/- 2.7 years; range: 18-29 years; 20 females, 9 males) using the Thermal Sensory Analyser TSA-II (Medoc Advanced Medical Systems, Ramat Yishai, Israel, and Minneapolis, Minnesota, USA) before and after laser needle acupuncture and placebo stimulation, respectively. Significant (p < or = 0,001; t-test) gender-specific differences were seen on cold pain threshold analysis. No significant changes in parameters of thermal sensory and pain thresholds were found before and after laser needle or placebo stimulation at acupuncture points for acute pain. However, a trend towards change in the median value of cold pain sensation after laser needle stimulation (p = 0.479; paired t-test; n.s.) was seen within the group of healthy females. The influence of stimulation of acupuncture points for chronic pain on the various parameters needs to be clarified in future studies.

Acupuncture↗

PubMed2003

Identification and quantitative expression analysis of genes that are differentially expressed during conidial germination in Pyrenophora teres.

Net blotch, caused by Pyrenophora teres, is a common disease of barley ( Hordeum vulgareL.). Two PCR-based differential screening techniques, cDNA-amplified fragment length polymorphism (cDNA-AFLP) and suppression subtractive hybridisation (SSH), were employed to clone cDNA copies of transcripts that are up-regulated during conidial germination. The nucleotide sequences of 35 transcripts were analysed, and the amino acid sequences of their predicted products were compared with entries in databases. Eleven of these clones showed homology to genes from other ascomycetes coding for a transcription factor, two regulatory proteins, a putative transposase, a protein required for the biogenesis of cytochrome C oxidase, a threonine synthase, a probable subunit of a phenylalanine-tRNA synthetase, a subunit of RNA polymerase I, a cation transport protein, a vacuolar ATP synthase subunit, and an RNA processing protein. One conserved hypothetical protein was found and 23 sequences could not be functionally classified. The relative expression of five transcripts at 0, 1, 2, 3, 6, 12 and 24 h after induction of germination was determined by real-time RT-PCR using 18S rRNA as the endogenous reference sequence. All transcripts showed a significant increase in expression during early stages of germination. The maximum change in expression relative to ungerminated conidia ranged between 2.6- and 6-fold. The characterisation of genes involved in biochemical processes during the germination of conidia could be useful for target-specific development of new antifungal agents.

Ascomycota↗

PubMed2002

Radiation doses of Swiss population from external sources.

The outdoor radiation exposure of the population in Switzerland from external sources results from cosmic background as well as from natural and artificial ground radiation. The geographical distribution of these components and of the total dose rate are represented on maps consisting of 2 kmx2 km grid cells. The average dose rate on Swiss territory outdoors is 147 nSv/h (1.29 mSv/a). The distributions are then related to the population density distribution by GIS application. The population is exposed to an average dose rate of 108 nSv/h (0.95 mSv/a) per capita which is just below the threshold for man-made dose rate given by national regulation. The lower value (relative to the country average 147 nSv/h) arises from the fact that most of the population lives north of the Alps where the lithology is dominated by rocks of relatively low radioactivity and where the cosmic radiation is low relative to the Alps.

Environmental Exposure↗

PubMed2002

Lamotrigine monotherapy for control of neuralgia after nerve section.

BACKGROUND: We present six patients treated only with the new-generation anticonvulsant lamotrigine to define its sole effect on neuralgia after nerve section. METHODS: Previous surgical or pharmacological attempts failed to relieve this neuropathic pain in our patients. Before initiation of lamotrigine therapy, patients reported spontaneous and touch-evoked shooting pain followed by periods of burning pain. No breakthrough medication was needed during the maintenance phase of 1-23 months. Data were acquired by a pain diary on a weekly basis. RESULTS: With 75-300 mg of lamotrigine per day, the burning and shooting pain intensity was relieved by 33-100%. Most obviously, the attack frequency of the shooting pain was reduced by 80-100%. No adverse effects were observed. CONCLUSION: We conclude that lamotrigine may be beneficial in the treatment of neuralgia after nerve section following the failure of previous pharmacological or surgical attempts.

Adult↗

PubMed2001

Identification and biochemical characterization of Arabidopsis thaliana sulfite oxidase. A new player in plant sulfur metabolism.

In mammals and birds, sulfite oxidase (SO) is a homodimeric molybdenum enzyme consisting of an N-terminal heme domain and a C-terminal molybdenum domain (EC ). In plants, the existence of SO has not yet been demonstrated, while sulfite reductase as part of sulfur assimilation is well characterized. Here we report the cloning of a plant sulfite oxidase gene from Arabidopsis thaliana and the biochemical characterization of the encoded protein (At-SO). At-SO is a molybdenum enzyme with molybdopterin as an organic component of the molybdenum cofactor. In contrast to homologous animal enzymes, At-SO lacks the heme domain, which is evident both from the amino acid sequence and from its enzymological and spectral properties. Thus, among eukaryotes, At-SO is the only molybdenum enzyme yet described possessing no redox-active centers other than the molybdenum. UV-visible and EPR spectra as well as apparent K(m) values are presented and compared with the hepatic enzyme. Subcellular analysis of crude cell extracts showed that SO was mostly found in the peroxisomal fraction. In molybdenum cofactor mutants, the activity of SO was strongly reduced. Using antibodies directed against At-SO, we show that a cross-reacting protein of similar size occurs in a wide range of plant species, including both herbacious and woody plants.

Amino Acid Sequence↗

PubMed2001

Crystal structures of human gephyrin and plant Cnx1 G domains: comparative analysis and functional implications.

The molybdenum cofactor (Moco) consists of a unique and conserved pterin derivative, usually referred to as molybdopterin (MPT), which coordinates the essential transition metal molybdenum (Mo). Moco is required for the enzymatic activities of all Mo-enzymes, with the exception of nitrogenase and is synthesized by an evolutionary old multi-step pathway that is dependent on the activities of at least six gene products. In eukaryotes, the final step of Moco biosynthesis, i.e. transfer and insertion of Mo into MPT, is catalyzed by the two-domain proteins Cnx1 in plants and gephyrin in mammals. Gephyrin is ubiquitously expressed, and was initially found in the central nervous system, where it is essential for clustering of inhibitory neuroreceptors in the postsynaptic membrane. Gephyrin and Cnx1 contain at least two functional domains (E and G) that are homologous to the Escherichia coli proteins MoeA and MogA, the atomic structures of which have been solved recently. Here, we present the crystal structures of the N-terminal human gephyrin G domain (Geph-G) and the C-terminal Arabidopsis thaliana Cnx1 G domain (Cnx1-G) at 1.7 and 2.6 A resolution, respectively. These structures are highly similar and compared to MogA reveal four major differences in their three-dimensional structures: (1) In Geph-G and Cnx1-G an additional alpha-helix is present between the first beta-strand and alpha-helix of MogA. (2) The loop between alpha 2 and beta 2 undergoes conformational changes in all three structures. (3) A beta-hairpin loop found in MogA is absent from Geph-G and Cnx1-G. (4) The C terminus of Geph-G follows a different path from that in MogA. Based on the structures of the eukaryotic proteins and their comparisons with E. coli MogA, the predicted binding site for MPT has been further refined. In addition, the characterized alternative splice variants of gephyrin are analyzed in the context of the three-dimensional structure of Geph-G.

Alternative Splicing↗

PubMed2001

Thiocarboxylation of molybdopterin synthase provides evidence for the mechanism of dithiolene formation in metal-binding pterins.

Molybdopterin (MPT) is a pyranopterin with a unique dithiolene group coordinating molybdenum (Mo) or tungsten (W) in all Mo- and W-enzymes except nitrogenase. In Escherichia coli, MPT is formed by incorporation of two sulfur atoms into precursor Z, which is catalyzed by MPT synthase. The recently solved crystal structure of MPT synthase (Rudolph, M. J., Wuebbens, M. M., Rajagopalan, K. V., and Schindelin, H. (2000) Nat. Struct. Biol. 8, 42-46) shows the heterotetrameric nature of the enzyme that is composed of two small (MoaD) and two large subunits (MoaE). According to sequence and structural similarities among MoaD, ubiquitin, and ThiS, a thiocarboxylation of the C terminus of MoaD is proposed that would serve as the source of sulfur that is transferred to precursor Z. Here, we describe the in vitro generation of carboxylated and thiocarboxylated MoaD. Both forms of MoaD are monomeric and are able to form a heterotetrameric complex after coincubation in equimolar ratios with MoaE. Only the thiocarboxylated MPT synthase complex was found to be able to convert precursor Z in vitro to MPT. Slight but significant differences between the carboxylated and the thiocarboxylated MPT synthase can be seen using size exclusion chromatography. A two-step reaction of MPT synthesis is proposed where the dithiolene is generated by two thiocarboxylates derived from a single tetrameric MPT synthase.

Carboxylic Acids↗

PubMed2001

Negative cooperativity and aggregation in biphasic binding of mastoparan X peptide to membranes with acidic lipids.

The change of Trp fluorescence intensity when large vesicles with 10% acidic lipid are added to mastoparan X solutions reflects a fast and a slow binding process. By means of a novel procedure of data analysis that takes advantage of so-called mass conservation plots we have separated association isotherms related to: (i) the apparent fast pre-equilibrium; and (ii) the final equilibrium, respectively. This approach also reveals that the intrinsic fluorescence signal of the slow binding is considerably raised against that of the fast binding, presumably indicating a penetration of bound peptide from the lipid/water interface into the apolar lipid core. The shape of either binding curve discloses a pronounced tendency of aggregation. Furthermore, it turns out that in the slow process the final binding ratio decreases markedly compared with the initial fast binding ratio. Accordingly the occupation of final binding sites must exert a substantial effect of negative cooperativity on the affinity of the interfacial binding states.

Acids↗

PubMed2001

Pre-inoculation enrichment procedure enhances the performance of bacteriocinogenic Lactococcus lactis meat starter culture.

Sodium nitrite and sodium chloride may inhibit growth and bacteriocinogenesis of protective starter cultures. To reduce sensitivity of a lacticin 3147-producing starter culture to nitrite, prior to production of salami, Lactococcus lactis DPC 4275 was placed in a number of pre-inoculation treatments, containing (a) 1% glucose, (b) 2.5 ppm manganese (Mn), (c) 250 ppm magnesium (Mg), (d) 2.5 ppm manganese + 250 ppm magnesium (Mn + Mg), and held at ambient temperature for 30 min and 4 degrees C for 2 h. The growth, pH reduction, and bacteriocin production was monitored in beaker sausage over a period of 10 days at 28 degrees C, corresponding to typical salami production time, and compared to untreated starter culture. The effect of 1% tryptone and inoculum level on growth and bacteriocin production was also determined. Challenge tests were performed using Listeria innocua DPC 1770 and Staphylococcus aureus MMPR3 as target strains. All treatments gave a significantly higher (P < 0.05) initial starter level than the untreated starter. Beaker sausage inoculated with either low (10(7)) or high (10(9)) levels of starter culture, treated with Mn + Mg reached significantly (P < 0.05) higher levels by day 10 than other treatments. Trends indicate that Mn + Mg also gave best pH reduction in sausage containing the low-level starter culture, sausage and significantly lower (P < 0.05) values for sausage produced with higher inoculum. Bacteriocin production was also higher in starter culture treated with Mn, or glucose. Pre-treatment with Mg gave a 2-fold increase in bacteriocin, the addition of Mn augmenting this increase further. The incorporation of tryptone gave no additional effect. In beaker sausage, both L. innocua and S. aureus populations showed significant reductions (P < 0.05) in the presence of the bacteriocinogenic strain compared to a non-bacteriocinogenic control strain.

Antibiosis↗

PubMed2001

Cathepsin S and an asparagine-specific endoprotease dominate the proteolytic processing of human myelin basic protein in vitro.

The biochemical characterization of antigen degradation is an important basis for a better understanding of both the immune response and autoimmune diseases mediated by MHC class II molecules. In this study we used high-performance liquid chromatography and mass spectrometry to analyze the processing of myelin basic protein (MBP), a potential autoantigen implicated in the pathogenesis of multiple sclerosis. We resolved the kinetics of MBP processing by lysosomal extracts or purified endocytic proteases, identified the major cleavage sites during this process and assigned them to the activity of proteolytic enzymes. Proteolytic processing of MBP is mostly guided along the hydrophobic regions of the protein. It is initiated by two proteolytic steps (after N(92) and S(110)) that are performed by an asparagine-specific endopeptidase (AEP) and by cathepsin (Cat) S, respectively. The resulting processing intermediates are converted into more than 60 different species of 20-40-mers due to the activity of endopeptidases including CatS, D and L. The fragments thus generated are subsequently degraded by C- or N-terminal trimming. Strikingly, the initial cleavages during MBP processing affect two immunodominant regions of the potential autoantigen [MBP(85-99) and MBP(111-129)] in an inverse manner. CatS directly generates the N terminus of the epitope MBP(111-129) in large quantities during the initial phase of processing, which might explain the immunogenicity of this region in spite of its relatively poor binding to HLA-DR4. In contrast, the dominant cleavage by AEP mediates the destruction of MBP(85-99) unless the epitope is protected, e.g. by binding to HLA-DR. Our results thus characterize the proteolytic events during processing of MBP on a molecular level and suggest a biochemical basis for the immunogenicity of the immunodominant epitopes, which could serve as a guideline for future therapeutic strategies.

Amino Acid Sequence↗

PubMed2001

Laser doppler flowmetry in combined needle acupuncture and moxibustion: a pilot study in healthy adults.

Laser Doppler flowmetry (LDF) (DRT 4/Moor Instruments Ltd, Devon, UK) was used in this pilot study for monitoring the effects of an invariable acupuncture pattern on microcirculation of the skin before, during, and after combined needle acupuncture and moxibustion in 12 healthy volunteers (mean age 35.2 +/- 4.4 years, range 26-41 years, four female and eight male). According to the standards of traditional Chinese medicine (TCM), this acupuncture pattern is assumed to be unspecific and non-therapeutic. Flux decreased during the treatment period (p<0.05) compared to the control phase before combined needle acupuncture and moxibustion. After the removal of the needles, flux did not return to the initial control value. No significant differences between mean arterial blood pressure, heart rate, skin temperature or gender were detected. A prediction of individual incidences and kinds of acupuncture effects in our healthy volunteers was not possible. Modern monitoring techniques like LDF could be a method to separate responders from non-responders to acupuncture in peripheral microcirculatory disorders. Further studies on patients with peripheral microcirculatory disorders are necessary in order to demonstrate the value of LDF in detecting responders/non-responders in combination with therapeutic acupuncture patterns according to TCM.

Acupuncture↗

PubMed2001

Characterisation of the mob locus of Rhodobacter sphaeroides WS8: mobA is the only gene required for molybdopterin guanine dinucleotide synthesis.

The mob genes of several bacteria have been implicated in the conversion of molybdopterin to molybdopterin guanine dinucleotide. The mob locus of Rhodobacter sphaeroides WS8 comprises three genes, mobABC. Chromosomal in-frame deletions in each of the mob genes have been constructed. The mobA mutant strain has inactive DMSO reductase and periplasmic nitrate reductase activities (both molybdopterin guanine dinucleotide-requiring enzymes), but the activity of xanthine dehydrogenase, a molybdopterin enzyme, is unaffected. The inability of a mobA mutant to synthesise molybdopterin guanine dinucleotide is confirmed by analysis of cell extracts of the mobA strain for molybdenum cofactor forms following iodine oxidation. Mutations in mobB and mobC are not impaired for molybdoenzyme activities and accumulate wild-type levels of molybdopterin and molybdopterin guanine dinucleotide, indicating they are not compromised in molybdenum cofactor synthesis. In the mobA mutant strain, the inactive DMSO reductase is found in the periplasm, suggesting that molybdenum cofactor insertion is not necessarily a pre-requisite for export.

Cloning, Molecular↗

PubMed2001

Chain length dependence of lipid partitioning between the air/water interface and its subphase: thermodynamic and structural implications.

We have investigated phosphatidylcholines with the same two saturated hydrocarbon chains of 12, 10 and 8 C-atoms. Langmuir trough data could be evaluated towards even small lipid subphase desorption when applying a novel approach that had recently been developed in our laboratory. The C12 lipid turned out to form a nearly insoluble monolayer with slight desorption only beyond 15 mN/m for an area/volume ratio around 1 cm(-1). Above 22 mN/m micellation in the subphase apparently terminates further accumulation in the interface forcing additionally added lipid to enter the bulk volume. A comparatively substantial increase of solubilization was observed for the C10 monolayer. When turning to the C8 lipid partitioning proved to take place in nearly equal parts. In that case, strong multimeric aggregation is indicated to occur in both the interfacial and the bulk volume domains. All the results are quantitatively discussed in the light of basic thermodynamic and structural considerations.

Linear Models↗

PubMed2001

Reproducibility of HTS-SQUID magnetocardiography in an unshielded clinical environment.

A new technology has been developed which measures the magnetic field of the human heart (magnetocardiogram, MCG) by using high temperature superconducting (HTS) sensors. These sensors can be operated at the temperature of liquid nitrogen without electromagnetic shielding. We tested the reproducibility of HTS-MCG measurements in healthy volunteers. Unshielded HTS-MCG measurements were performed in 18 healthy volunteers in left precordial position in two separate sessions in a clinical environment. The heart cycles of 10 min were averaged, smoothed, the baselines were adjusted, and the data were standardized to the respective areas under the curves (AUC) of the absolute values of the QRST amplitudes. The QRS complexes and the ST-T intervals were used to assess the reproducibility of the two measurements. Ratios (R(QRS), R(STT)) were calculated by dividing the AUC of the first measurement by the ones of the second measurement. The linear correlation coefficients (CORR(QRS), CORR(STT)) of the time intervals of the two measurements were calculated, too. The HTS-MCG signal was completely concealed by the high noise level in the raw data. The averaging and smoothing algorithms unmasked the QRS complex and the ST segment. A high reproducibility was found for the QRS complex (R(QRS)=1.2+/-0.3, CORR(QRS)=0.96+/-0.06). Similarly to the shape of the ECG it was characterized by three bends, the Q, R, and S waves. In the ST-T interval, the reproducibility was considerably lower (R(STT)=0.9+/-0.2, CORR(STT)=0.66+/-0.28). In contrast to the shape of the ECG, a baseline deflection after the T wave which may belong to U wave activity was found in a number of volunteers. HTS-MCG devices can be operated in a clinical environment without shielding. Whereas the reproducibility was found to be high for the depolarization interval, it was considerably lower for the ST segment and for the T wave. Therefore, before clinically applying HTS-MCG systems to the detection of repolarization abnormalities in acute coronary syndromes, further technical development of the systems is necessary to improve the signal-to-noise ratio.

Adult↗