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Biomedical subjects

G Schwarz

Publications and source records attributed to G Schwarz.

At least 55 records · Page 3Linked to original sources
PubMed2000

The molybdenum cofactor biosynthetic protein Cnx1 complements molybdate-repairable mutants, transfers molybdenum to the metal binding pterin, and is associated with the cytoskeleton.

Molybdenum (Mo) plays an essential role in the active site of all eukaryotic Mo-containing enzymes. In plants, Mo enzymes are important for nitrate assimilation, phytohormone synthesis, and purine catabolism. Mo is bound to a unique metal binding pterin (molybdopterin [MPT]), thereby forming the active Mo cofactor (Moco), which is highly conserved in eukaryotes, eubacteria, and archaebacteria. Here, we describe the function of the two-domain protein Cnx1 from Arabidopsis in the final step of Moco biosynthesis. Cnx1 is constitutively expressed in all organs and in plants grown on different nitrogen sources. Mo-repairable cnxA mutants from Nicotiana plumbaginifolia accumulate MPT and show altered Cnx1 expression. Transformation of cnxA mutants and the corresponding Arabidopsis chl-6 mutant with cnx1 cDNA resulted in functional reconstitution of their Moco deficiency. We also identified a point mutation in the Cnx1 E domain of Arabidopsis chl-6 that causes the molybdate-repairable phenotype. Recombinant Cnx1 protein is capable of synthesizing Moco. The G domain binds and activates MPT, whereas the E domain is essential for activating Mo. In addition, Cnx1 binds to the cytoskeleton in the same way that its mammalian homolog gephyrin does in neuronal cells, which suggests a hypothetical model for anchoring the Moco-synthetic machinery by Cnx1 in plant cells.

Amino Acid Sequence↗
PubMed2000

Epstein-Barr virus infection in peripheral blood mononuclear cells, synovial fluid cells, and synovial membranes of patients with rheumatoid arthritis.

OBJECTIVE: Different infectious agents have been proposed to be involved in the pathogenesis of rheumatoid arthritis (RA). We investigated the role of latent Epstein-Barr virus (EBV) infection in patients with RA. METHODS: Sera of 55 patients with RA and 60 age and sex matched healthy controls were examined for antibodies against EBV encoded antigens (EBNA-1, VCA, and EA) by ELISA. The presence of viral DNA was analyzed by polymerase chain amplification of DNA isolated from peripheral blood mononuclear cells (PBMC) of these samples and from cells of synovial fluid (SF) specimens. Paraffin sections of synovial membranes from 25 patients were subjected to in situ hybridization analysis for the EBV encoded small RNA EBER1 and EBER2. RESULTS: Two-fold increased values of IgG antibodies against EBNA-1 were found in patients with RA in comparison to healthy controls (p = 0.029). No statistically significant difference could be observed for antibody levels against EBV-VCA. Fourteen (24.1%) of 55 patients with RA had serological evidence of reactivated EBV infection in comparison to none of the control group (p = 0.028). In PBMC, EBV DNA was detected in a significantly higher proportion in the patient group (50.9 vs. 30%; p = 0.02). In addition, SF cells harbored the viral DNA in 30% of RA cases compared to 16.6% of control cases (p = 0.02). However, EBER1/2 transcripts could only be found within synovial membranes of 2 (8%) of 25 patients with RA. CONCLUSION: These findings support the hypothesis that EBV infection may be involved in the pathogenesis of RA. Further studies may define the precise pathogenetic mechanisms of viral infection for the development of inflammatory arthritis.

Adolescent↗
PubMed1999

Pore-forming action of mastoparan peptides on liposomes: a quantitative analysis.

We have investigated the wasp venom peptides mastoparan X and polistes mastoparan regarding their apparent potential to induce pore-like defects in phosphatidylcholine unilamellar vesicles. Based on a fundamental theoretical model, the pore activation and deactivation kinetics have been evaluated from the observed efflux of liposome entrapped carboxyfluorescein in relation to the bound peptide to lipid ratio. We can quantitatively describe our experimental data very well in terms of a specific reaction scheme resulting in only a few short-lived pores. They evidently emerge rapidly from a prepore nucleus being produced by two rate-limiting monomeric states of bound peptide. These peculiar states would be favorably populated in an early stage of bilayer perturbation, but tend to die out in the course of a peptide/lipid restabilization process.

Amino Acid Sequence↗
PubMed1999

The neurotransmitter receptor-anchoring protein gephyrin reconstitutes molybdenum cofactor biosynthesis in bacteria, plants, and mammalian cells.

The molybdenum cofactor (Moco), a highly conserved pterin compound complexing molybdenum, is required for the enzymatic activities of all molybdenum enzymes except nitrogenase. Moco is synthesized by a unique and evolutionarily old pathway that requires the activities of at least six gene products. Some of the proteins involved in bacterial, plant, and invertebrate Moco biosynthesis show striking homologies to the primary structure of gephyrin, a polypeptide required for the clustering of inhibitory glycine receptors in postsynaptic membranes in the rat central nervous system. Here, we show that gephyrin binds with high affinity to molybdopterin, the metabolic precursor of Moco. Furthermore, gephyrin expression can reconstitute Moco biosynthesis in Moco-deficient bacteria, a molybdenum-dependent mouse cell line, and a Moco-deficient plant mutant. Conversely, inhibition of gephyrin expression by antisense RNA expression in cultured murine cells reduces their Moco content significantly. These data indicate that in addition to clustering glycine receptors, gephyrin also is involved in Moco biosynthesis and illustrate the remarkable conservation of its function in Moco biosynthesis throughout phylogeny.

Animals↗
PubMed1999

Polymorphism and interactions of a viral fusion peptide in a compressed lipid monolayer.

With a view toward possible new insights into viral fusion mechanisms, we have investigated the HIV-1 gp41 fusion peptide in a monomolecular film of the biomembrane lipid palmitoyloleoylphosphatidylcholine. Its surface activity at an air/water interface was measured under equilibrium conditions, using the conventional Langmuir trough technique. Through a novel thermodynamic analysis, the partial molecular area of the peptide in the lipid moiety could be determined as a function of the lateral pressure and the interfacial peptide/lipid ratio. This indicates an orientation of the peptide backbone parallel to the lipid hydrocarbon tails. The molecular area decreases significantly upon monolayer compression, suggesting a conformational transition from a somewhat compact configuration to a more extended, presumably beta-strand structure when a lipid packing density is approached that is generally believed to mimic the physical state of a biological membrane. Up to a lateral pressure of approximately 15 mN/m, practically all peptide inserts into the lipid monolayer. At higher compression a distinct partitioning into the aqueous subphase is observed. Under these conditions the data also reflect a strong aggregation of the lipid-associated peptide. Beyond a critical peptide/lipid ratio, the peptide's area requirement was found to become substantially enhanced, possibly because of the formation of water-filled pores.

Amino Acid Sequence↗
PubMed1999

Ultrasound-monitored effects of acupuncture on brain and eye.

A new transcranial Doppler sonography arrangement was used to monitor blood flow profiles in the supratrochlear and middle cerebral arteries simultaneously and continuously. The technique selectively demonstrated the specific effect of acupuncture on the cranial arteries, in a 25-year-old female with pigmentary retinopathy. Stimulation of points Zanzhu and Yuyao led to a marked increase of blood flow velocity in the supratrochlear artery and to a decrease of flow velocity, in the middle cerebral artery. These acupuncture-induced effects were reproducible even though both arteries originate from the same major vessel.

Acupuncture Therapy↗
PubMed1999

Computer-controlled acupuncture. Quantification and separation of specific effects.

The effects of acupuncture have been described and handed down empirically for centuries but there are few reports based on objective data. The aim of this study was to provide selective evidence of a specific effect of acupuncture on the brain and the eye using a Doppler ultrasound technique. A transcranial Doppler sonography arrangement was developed to monitor blood flow profiles in the supratrochlear and middle cerebral arteries simultaneously and continuously. Two acupuncture schemas were tested in a randomized cross-over study with 13 patients with ophthalmologic diseases. Applying acupuncture needles to special eye points increased the blood flow velocity in the supratrochlear artery significantly (p < 0.001) compared to the reference interval before acupuncture. In the middle cerebral artery only a minimal, non-significant increase in blood flow velocity was seen. In contrast, acupuncture of points that are held to increase cerebral blood flow velocity increased blood flow velocity in the middle cerebral artery significantly (p < 0.001) while leaving that in the supratrochlear artery unchanged. Specific acupuncture produce specific, reproducible quantifiable effects on blood flow velocity in arteries to the brain and eye.

Acupuncture Therapy↗
PubMed1999

Galectin fingerprinting in tumor diagnosis. Differential expression of galectin-3 and galectin-3 binding sites, but not galectin-1, in benign vs malignant uterine smooth muscle tumors.

Cell-matrix interactions are governed by a distinct set of proteins, with 2 nonintegrin laminin-binding proteins, galectin-1 and galectin-3, providing 1 aspect. The expression patterns of laminin and the 2 galectins and galectin binding sites were quantitatively determined by means of computer-assisted microscopy with the aim of differentiating between 16 leiomyomas and 10 leiomyosarcomas of the uterus. Three quantitative variables were computed for each of the 5 histochemical markers: labeling index, which describes the percentage of tissue area specifically stained by a given marker; mean optical density which reflects the concentration of the marker; and concentrational heterogeneity, which characterizes the degree of heterogeneity of the marker distribution in the tumor tissue areas. The results reveal evident differences in the galectin-3-related parameters in the 2 tumors groups. Whereas the concentration of galectin-3 binding sites was significantly (P = .01) weaker in the leiomyosarcomas than in the leiomyomas, the percentages of tumor tissue expressing galectin-3 (P = .02) and its binding sites (P = .002) were significantly higher in the leiomyosarcomas than in the leiomyomas. Although significantly (P = .02) higher, the concentration of laminin was more heterogeneously distributed (P = .01) in the leiomyosarcomas than in the leiomyomas. In contrast, the levels of expression of galectin-1 and its accessible binding sites remained similar for both the leiomyomas and the leiomyosarcomas. Finally we document how the levels of expression of galectin-3 and its binding sites can be of assistance in reliably differentiating leiomyomas from leiomyosarcomas.

Adult↗
PubMed1999

Fiberoptic intubation in 327 neurosurgical patients with lesions of the cervical spine.

In patients with lesions of the cervical spine, direct laryngoscopy for endotracheal intubation entails the risk of injuring the spinal cord. In an attempt to avoid this complication, the authors used flexible fiberoptic nasal intubation in a series of 327 patients with cervical lesions undergoing elective neurosurgical procedures. The nasal route was preferred for laryngeal intubation because it is easier than the oral route and a restraining collar or halo device does not impair the intubating maneuver. Bronchoscopic intubation was possible in all patients. In 12 patients (3.6%), anatomic abnormalities prevented transnasal insertion of the endotracheal tube, and transoral fiberoptic intubation was necessary. Endotracheal intubation was graded as slightly difficult in 85 patients (26%). The minimal peripheral oxygen saturation during intubation exceeded 90% in 289 patients (88%). In the other 38 patients, the mean O2 saturation was 84.2+/-4.3% (range, 72-89%). Intubation was well tolerated by all patients and none had recall of the procedure. Cervical stabilizers did not have to be removed for intubation in any patient. None of the patients had postoperative neurologic deficits attributable to the intubation procedure. The authors consider fiberoptic transnasal intubation to be a useful alternative to direct laryngoscopic tracheal intubation in patients undergoing elective surgical procedures on the cervical spine to avoid potential injury to the cervical spinal cord.

Adolescent↗
PubMed1999

Is there paradoxical arousal reaction in the EEG subdelta range in patients during anesthesia?

Different anesthetic drugs can produce different electroencephalographic (EEG) patterns. Nondrug induced influences, such as surgical stimulation, may also alter the EEG in anesthetized patients. Increases of delta activity are interpreted as signs of deepening of anesthesia. These EEG patterns are sometimes also found in frontal leads after surgical stimulation during isoflurane anesthesia. However, to avoid incorrect anesthetic management under EEG monitoring, this phenomenon of "paradoxical arousal" must be taken into consideration. This study demonstrates that, after surgical stimulation, frontal activity was enhanced in the 2-3 Hz and 3-4 Hz frequency bands. On the other hand, in the subdelta range (the 0.05-1 Hz and 1-2 Hz frequency bands), there was an opposite reaction. This significant (p<0.001, according to ANOVA and Dunn's method) reduction of EEG power as a result of the surgical stimuli was found in 25 patients (mean age, 49.1+/-16.3 years) scheduled for elective neurosurgical intervention in the lumbosacral region. This report shows some of the problems arising from the common practice of arbitrarily subdividing a power spectrum in frequency bands.

Adolescent↗
PubMed1999

Activity of the molybdopterin-containing xanthine dehydrogenase of Rhodobacter capsulatus can be restored by high molybdenum concentrations in a moeA mutant defective in molybdenum cofactor biosynthesis.

During the screening for Rhodobacter capsulatus mutants defective in xanthine degradation, one Tn5 mutant which was able to grow with xanthine as a sole nitrogen source only in the presence of high molybdate concentrations (1 mM), a phenotype resembling Escherichia coli mogA mutants, was identified. Unexpectedly, the corresponding Tn5 insertion was located within the moeA gene. Partial DNA sequence analysis and interposon mutagenesis of regions flanking R. capsulatus moeA revealed that no further genes essential for molybdopterin biosynthesis are located in the vicinity of moeA and revealed that moeA forms a monocistronic transcriptional unit in R. capsulatus. Amino acid sequence alignments of R. capsulatus MoeA (414 amino acids [aa]) with E. coli MogA (195 aa) showed that MoeA contains an internal domain homologous to MogA, suggesting similar functions of these proteins in the biosynthesis of the molybdenum cofactor. Interposon mutants defective in moeA did not exhibit dimethyl sulfoxide reductase or nitrate reductase activity, which both require the molybdopterin guanine dinucleotide (MGD) cofactor, even after addition of 1 mM molybdate to the medium. In contrast, the activity of xanthine dehydrogenase, which binds the molybdopterin (MPT) cofactor, was restored to wild-type levels after the addition of 1 mM molybdate to the growth medium. Analysis of fluorescent derivatives of the molybdenum cofactor of purified xanthine dehydrogenase isolated from moeA and modA mutant strains, respectively, revealed that MPT is inserted into the enzyme only after molybdenum chelation, and both metal chelation and Mo-MPT insertion can occur only under high molybdate concentrations in the absence of MoeA. These data support a model for the biosynthesis of the molybdenum cofactor in which the biosynthesis of MPT and MGD are split at a stage when the molybdenum atom is added to MPT.

Amino Acid Sequence↗
PubMed1999

[Computer-controlled acupuncture. A new construction for simultaneous measurement of blood flow velocity of the supratrochlear and middle cerebral arteries].

Acupuncture has been used for over 5,000 years and a large body of evidence exists. Transcranial Doppler (TCD) ultrasound, introduced more than a decade ago, has evolved into a dynamic, reliable, reproducible and practical diagnostic tool. In this study we describe simultaneous and continuous TCD monitoring of the supratrochlear and middle cerebral arteries in two patients with ophthalmologic diseases undergoing computer-controlled acupuncture (CCA). During eye acupuncture scheme there was a significant increase in the mean bloodflow velocity (vm) of the A. supratrochlearis (+39% patient 1; +15% patient 2; p < 0.001, t-test) in both patients. The effect was still evident after the needles were removed. There was a smaller increase (+4% and +3%; n.s.) of vm in the A. cerebri media. These results suggest that eye acupuncture could have a specific effect on intra- and extracerebral arteries. CCA might improve the quality of acupuncture.

Acupuncture Therapy↗
PubMed1998

Use of a broad-host-range bacteriocin-producing Lactococcus lactis transconjugant as an alternative starter for salami manufacture.

Lactococcus lactis DPC4268 is widely used for Cheddar cheese manufacture in Ireland where it is recognised for its reliable fast acid producing ability in dairy environments. A transconjugant of this strain, L. lactis DPC4275, was generated which produces the broad spectrum, two-component bacteriocin lacticin 3147, which is inhibitory to a variety of undesirable gram-positive bacteria including Clostridium, Bacillus, Enterococcus, Listeria, Staphylococcus and Streptococcus. Both DPC4268 and DPC4275 were used as single strain starters for manufacture of salamis. These products were compared with a salami manufactured with a conventional starter (Lactobacillus sake and Staphylococcus carnosus) in terms of pH development, a(w)-value, weight loss, colour development and sensory characteristics. Salamis produced with either lactococcal culture exhibited pH values below 5.1 and a(w)-values below 0.90 which is favourable for preservation and hygienic stability. In addition, these salamis had good sensory and colorimetric qualities. A minimum lacticin 3147 concentration of 640 AU/g was detected in the salamis which were produced with L. lactis DPC4275.

Animals↗
PubMed1998

The Chlamydomonas reinhardtii MoCo carrier protein is multimeric and stabilizes molybdopterin cofactor in a molybdate charged form.

In Chlamydomonas reinhardtii, molybdopterin cofactor (MoCo) able to reconstitute active nitrate reductase (NR) with apoenzyme from the Neurospora crassa mutant nit-1 was found mostly bound to a carrier protein (CP). This protein is scarce in the algal free extracts and has been purified 520-fold. MoCoCP is a protein of 64 kDa with subunits of 16.5 kDa and an isoelectric point of 4.5. In contrast to free MoCo, MoCo bound to CP was remarkably protected against inactivation under both aerobic conditions and basic pH. MocoCP transferred active MoCo to apoNR in vitro without addition of molybdate, though reconstituted activity was 20% higher in the presence of molybdate. Incubation with tungstate specifically inhibited MoCoCP activity but had no effect on the activity of free MoCo released from milk xanthine oxidase. MoCoCP did not charge molybdate unless in the presence of N. crassa extracts. Our data support that MoCoCP stabilizes MoCo in an active form charged with molybdate to provide MoCo to apomolybdoenzymes.

Animals↗
PubMed1998

Peptide-liposome association. A critical examination with mastoparan-X.

Mastoparan-X, a wasp venom factor, is a membrane active peptide whose binding to lipid vesicles is of basic interest towards an analysis of its functions. Titration of aqueous peptide solutions with liposomes allows the determination of the association isotherm, i.e. a plot of bound peptide per lipid versus the free peptide concentration. We have scrutinised the various steps in the evaluation procedure, considering circular dichroism as well as fluorescence intensity as possible signals for the binding process. First of all the measured data had to be corrected for light scattering effects which may otherwise appreciably falsify the final results. Uncertainties due to inherent difficulties regarding the reproducibility of lipid preparations and inevitable titration errors have to be considered. The consequences of these errors for the quantitative analysis of the titration curves were investigated. The plotted curves can be reasonably well fitted by a functional relationship derived from a Gouy-Chapman model approach that assumes a partitioning of monomeric peptide. The two relevant parameters, partition coefficient and effective charge number, and their error ranges have been determined for mastoparan-X and a series of phosphatidylcholine vesicle sizes and various ionic strengths. These findings show that the applied analysis implies a sufficient basis for calculations of the amount of lipid bound peptide in practice. However, the possible existence of peptide aggregates cannot generally be excluded from a formal monomer associated curve fit as indicated by computer simulations.

Circular Dichroism↗
PubMed1998

Quantitative studies on the melittin-induced leakage mechanism of lipid vesicles.

We have investigated, both experimentally and theoretically, the efflux of carboxyfluorescein (a self-quenching fluorescent dye) from vesicles of different sizes and lipid species (POPC, DOPC) after having added the bee venom peptide melittin. This comprises quantitative analyses regarding the extent of lipid-associated peptide, the mode as well as the temporal progress of dye release and the possible leakage mechanism. Our results indicate a graded efflux characterized by a single-pore retention factor reflecting the formation of pores whose lifetimes are rather small (millisecond range). The observed fluorescence signal arising from the dequenching of effluent dye has been converted to the number of pore openings over the course of time. All the resulting curves exhibit a pronounced slowing down of the pore formation rate revealing two distinct relaxation steps at about 20 and 200 s, respectively, being largely independent of vesicle type and peptide to lipid ratio. The pore formation rate itself increases in proportion to the amount of membrane bound peptide. We give a quantitative account of our experimental findings based on a novel reaction scheme applicable to any of our various liposome systems. It implies that the pore formation rate is controlled by a passage through two intermediate monomeric peptide states. These states are thought to become well populated in the initial stage of lipid bilayer perturbation, but would practically die out after some time owing to a restabilization of the membrane system.

Binding Sites↗