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Biomedical subjects

G Wagner

Publications and source records attributed to G Wagner.

At least 73 records · Page 4Linked to original sources
PubMed2001

[Value of several examination systems in patients with carpal tunnel syndrome. Comparison of Dellon computer-assisted sensation test with Mellesi hand status and Levine examination scheme].

The goal of the presented carpal tunnel syndrome (CTS) follow-up study was to compare the clinical value of the Millesi hand function score with the "Pressure Specifying Sensory Device" (PSSD) introduced by A. L. Dellon using self-administered patient questionnaires. 25 patients (10 male, 15 female) with an electrodiagnostically confirmed CTS were enrolled in this study, performing one preoperative and five postoperative examinations over 24 weeks. 12 of the 25 patients underwent an "open" two-portal carpal tunnel release with two minimal incisions (group OT); the other 13 patients were treated with a two-portal endoscopic carpal tunnel release (group ET). Additionally, in eleven out of the twelve patients of group OT, an epineuriotomy of the median nerve was performed during the same session. Concerning preoperative data of the Millesi score and the PSSD, no statistically significant differences were found between group OT and ET. However, preoperative comparison with the contralateral hand demonstrated a reduction in hand function of 15% and an increase in the pressure perception threshold of 41% compared to normative data could be measured with the PSSD. The subjective functional value of the hand was objectively evacuated using the Levine score. In the second postoperative week, a significant decrease in hand function could be obtained with the Millesi score in group OT. In group ET, the decrease in hand function representing the operative trauma was significantly lower than in group OT. Data of the static one- and two-point pressure perception threshold revealed a statistically significant improvement of the sensibility in both groups. The results of the Millesi score recorded at the last examination in the 24th postoperative week showed an improvement in hand function in both groups compared to preoperative data. Concerning static one- and two-point measurements with the PSSD, distinct improvements compared to the preoperative data could also be detected in both groups although significant differences between group OT and ET were evident: Data of group OT regarding the whole postoperative course demonstrate a continuous improvement in sensibility of the index finger. In contrast, the analysis of the PSSD measurements in group ET revealed an increase in all the parameters starting in the sixth postoperative week and ending with significantly worse static one- and two-point threshold measurements than in group OT. On the other hand, data of self-administered patient questionnaires using the Levine Score revealed significant improvements in hand function and reduction in pain intensity in both groups compared to preoperative results. Differences between both groups at the end of the examination course were not evident. In conclusion, the Millesi hand score with its emphasis on the motor function proved to be a reliable method to record the severity of CTS preoperatively, the severity of the surgical trauma and changes in the course of rehabilitation of the affected hand. A good correlation was found between data obtained with the Millesi Score and the self-administered patient questionnaires according to Levine. But when compared with the PSSD, both methods could not directly document the preoperative status and postoperative changes of the median nerve. Preoperative static two-point pressure threshold measurements with the PSSD confirmed their status as a screening parameter as published by A. L. Dellon. In the postoperative course of group ET, a distinct worsening in the sensibility of the index and little finger could only be detected with the PSSD before the patients noticed the onset of related symptoms.

Adult↗
PubMed2001

Sildenafil citrate (VIAGRA) improves erectile function in elderly patients with erectile dysfunction: a subgroup analysis.

BACKGROUND: The prevalence of erectile dysfunction (ED) increases with advancing age, with a particularly high prevalence of ED in elderly patients with diabetes. In the United States it is estimated that approximately 45% of men aged 65 to 69 years have moderate or complete ED. The efficacy and safety of oral sildenafil (VIAGRA) for treating ED in elderly men (aged > or = 65 years or older) were assessed. METHODS: We analyzed data obtained from five double-blind, placebo-controlled studies of the efficacy and tolerability of oral sildenafil taken as required (but not more than once daily) over a 12-week to 6-month period. Two subgroups were evaluated: (i) elderly patients with ED of broad-spectrum etiology (n = 411), and (ii) elderly patients with ED and diabetes (n = 71). Efficacy was assessed using a global efficacy question, questions 3 and 4 of the International Index of Erectile Function (IIEF). and the five sexual function domains of the IIEF. RESULTS: All efficacy assessments indicated that sildenafil significantly improved erectile function both in elderly patients with ED of broad-spectrum etiology and in elderly patients with ED and diabetes. The most common adverse events were mild-to-moderate headache, flushing, and dyspepsia. The rates of discontinuation due to adverse events were low and were comparable to the rates with placebo. CONCLUSIONS: Sildenafil is an efficacious and well-tolerated treatment for ED in elderly men.

Aged↗
PubMed2001

Solution structure and backbone dynamics of an omega-conotoxin precursor.

Nuclear magnetic resonance spectroscopy was used to characterize the solution structure and backbone dynamics of a putative precursor form of omega-conotoxin MVIIA, a 25-amino-acid residue peptide antagonist of voltage-gated Ca(2+) channels. The mature peptide is found in the venom of a fish-hunting marine snail Conus magus and contains an amidated carboxyl terminus that is generated by oxidative cleavage of a Gly residue. The form examined in this study is identical to the mature peptide except for the presence of the unmodified carboxy-terminal Gly. This form, referred to as omega-MVIIA-Gly, has previously been shown to refold and form its disulfides more efficiently than the mature form, suggesting that the presence of the terminal Gly may favor folding in vivo. The nuclear magnetic resonance (NMR) structure determination indicated that the fold of omega-MVIIA-Gly is very similar to that previously determined for the mature form, but revealed that the terminal Gly residue participates in a network of hydrogen bonds involving both backbone and side chain atoms, very likely accounting for the enhanced stability and folding efficiency. (15)N relaxation experiments indicated that the backbone is well ordered on the nanosecond time scale but that residues 9-15 undergo a conformational exchange processes with a time constant of approximately 35 microseconds. Other studies have implicated this segment in the binding of the peptide to its physiological target, and the observed motions may play a role in allowing the peptide to enter the binding site

Calcium Channel Blockers↗
PubMed2001

Enhanced gamma-glutamyl transpeptidase expression and superoxide production in Mpv17-/- glomerulosclerosis mice.

Recently, gamma-glutamyl transpeptidase, which initiates cleavage of extracellular glutathione, has been shown to promote oxidative damage to cells. Here we examined a murine disease model of glomerulosclerosis, involving loss of the Mpv17 gene coding for a peroxisomal protein. In Mpv17-/- cells, enzyme activity and mRNA expression (examined by quantitative RT-PCR) of membrane-bound gamma-glutamyl transpeptidase were increased, while plasma glutathione peroxidase and superoxide dismutase levels were lowered. Superoxide anion production in these cells was increased as documented by electron spin resonance spectroscopy. In the presence of Mn(III)tetrakis(4-benzoic acid)porphyrin, the activities of gamma-glutamyl transpeptidase and plasma glutathione peroxidase were unchanged, suggesting a relationship between enzyme expression and the amount of reactive oxygen species. Inhibition of gamma-glutamyl transpeptidase by acivicin reverted the lowered plasma glutathione peroxidase and superoxide dismutase activities, indicating reciprocal control of gene expression for these enzymes.

Animals↗
PubMed2001

Incorporation of horseradish peroxidase in a Kieselguhr membrane and the application to a mediator-free hydrogen peroxide sensor.

Horseradish peroxidase was incorporated in a kieselguhr membrane. The electron-transfer process of the enzyme was examined by cyclic voltammetry. It was observed that the electron-transfer reactivity of horseradish peroxidase was greatly enhanced, and that direct electrochemistry was accordingly feasible. Using the merits of the direct electron-transfer reactivity of horseradish peroxidase and its specific enzymatic catalysis towards hydrogen peroxide, an unmediated hydrogen peroxide biosensor was constructed. The calibration plot of this hydrogen peroxide sensor was linear in the range of 2.0 x 10(-6) mol/L - 6.5 x 10(-4) mol/L. The relative standard deviation was 4.1% for 6 successive determinations at a concentration of 1.0 x 10(-4) mol/L. The detection limit was 1.0 x 10(-6) mol/L.

Biosensing Techniques↗
PubMed2001

Preparing the veterinary profession for corporate and trade issues in the Americas: proceedings of a conference on synergism and globalization, Santiago, Chile, May 6-8, 2001.

The complex and rapid-paced development of international trade, coupled with increasing societal demands for the production not only of abundant and inexpensive food, but also of food that is safe and has been raised in a humane and environmentally friendly manner, demands immediate attention from the veterinary community. The new culture of global trade agreements, spurred by the development of the WTO, dictates massive changes and increasing integration of public and private sectors. This is a huge growth area for our profession and will require individuals with a skill set we do not yet provide in our educational framework. In North America, veterinary education is parochial and focused on specialization. This strong orientation toward companion animals fails to provide adequate training for those interested in acquiring the necessary skills for the emerging area of globalization and trade. In South America, curricula are less harmonized with one another and there is tremendous variation in degree programs, rendering it difficult to ascertain whether veterinarians are prepared to assume decision-making responsibilities regarding international transport of food. If we do not begin to prepare our graduates adequately for this emerging market demand, the positions will be filled by other professions. These other professions lack broad-based scientific knowledge about animal physiology and disease causation. Decisions made without adequate background could have devastating consequences for society, including incursions of unwelcome diseases, food safety problems, and public health issues. To prepare our new veterinary graduates for the future and this emerging market, it is important to nurture a global mindset within our academic communities and to promote communications, languages, and an interdependent team mentality. Areas of technical expertise that need a place, perhaps a parallel track, in the curriculum include production medicine, public health, food safety, and international veterinary medicine. The major trade corridors of the future regarding animal-based protein flow between North and South America. It is absolutely essential that we find areas of consensus and deficiencies so that we can harmonize our trade agreements and ensure adequate flow of safe food products from one continent to the other.

Americas↗
PubMed2000

Changes in deep-water formation during the Younger Dryas event inferred from 10Be and 14C records.

Variations in atmospheric radiocarbon (14C) concentrations can be attributed either to changes in the carbon cycle--through the rate of radiocarbon removal from the atmosphere--or to variations in the production rate of 14C due to changes in solar activity or the Earth's magnetic field. The production rates of 10Be and 14C vary in the same way, but whereas atmospheric radiocarbon concentrations are additionally affected by the carbon cycle, 10Be concentrations reflect production rates more directly. A record of the 10Be production-rate variations can therefore be used to separate the two influences--production rates and the carbon cycle--on radiocarbon concentrations. Here we present such an analysis of the large fluctuations in atmospheric 14C concentrations, of unclear origin, that occurred during the Younger Dryas cold period. We use the 10Be record from the GISP2 ice core to model past production rates of radionuclides, and find that the largest part of the fluctuations in atmospheric radiocarbon concentrations can be attributed to variations in production rate. The residual difference between measured 14C concentrations and those modelled using the 10Be record can be explained with an additional change in the carbon cycle, most probably in the amount of deep-water formation.

Journal Article↗
PubMed2000

Differential stabilization of two hydrophobic cores in the transition state of the villin 14T folding reaction.

We report the distribution of hydrophobic core contacts during the folding reaction transition state for villin 14T, a small 126-residue protein domain. The solution structure of villin 14T contains a central beta-sheet with two flanking hydrophobic cores; transition states for this protein topology have not been previously studied. Villin 14T has no disulfide bonds or cis-proline residues in its native state; it folds reversibly, and in an apparently two-state manner under some conditions. To map the hydrophobic core contacts in the transition state, 27 point mutations were generated at positions spread throughout the two hydrophobic cores. After each point mutation, comparison of the change in folding kinetics with the equilibrium destabilization indicates whether the site of mutation is stabilized in the transition state. The results show that the folding nucleus, or the sub-region with the strongest transition state contacts, is located in one of the two hydrophobic cores (the predominantly aliphatic core). The other hydrophobic core, which is mostly aromatic, makes much weaker contacts in the transition state. This work is the first transition state mapping for a protein with multiple major hydrophobic cores in a single folding unit; the hydrophobic cores cannot be separated into individual folding subdomains. The stabilization of only one hydrophobic core in the transition state illustrates that hydrophobic core formation is not intrinsically capable of nucleating folding, but must also involve the right specific interactions or topological factors in order to be kinetically important.

Amino Acid Sequence↗
PubMed2000

Heterodimeric CD3epsilongamma extracellular domain fragments: production, purification and structural analysis.

The CD3 polypeptides (epsilon, gamma, and delta) are non-covalently associated signaling subunits of the T cell receptor which form non-disulfide linked epsilongamma and epsilondelta heterodimers. With the goal of investigating their structure, Escherichia coli expression was utilized to produce CD3 ectodomain fragments including the murine CD3epsilon subunit N-terminal Ig-like extracellular domain alone or as a single chain construct with that of CD3gamma. The latter links the CD3gamma segment to the C terminus of the CD3epsilon segment via a 26 amino acid peptide (scCD3epsilongamma26). Although CD3epsilon could be produced at high yield when directed to inclusion bodies, the refolded monomeric CD3epsilon was not native as judged by monoclonal antibody binding using surface plasmon resonance and was largely unstructured by (15)N-(1)H two-dimensional NMR analysis. In contrast, scCD3epsilongamma26 could be refolded readily into a native state as shown by CD, NMR and mAb reactivity. The linker length between CD3epsilon and CD3gamma is critical since scCD3epsilongamma16 containing a 16 residue connector failed to generate a stable heterodimer. Collectively, the results demonstrate that: (i) soluble heterodimeric fragments of CD3 can be produced; (ii) cotranslation of CD3 chains insures proper folding even in the absence of the conserved ectodomain stalk region (CxxCxE); and (iii) CD3epsilon has a more stable tertiary protein fold than CD3gamma.

Amino Acid Motifs↗
PubMed2000

SH3 domain recognition of a proline-independent tyrosine-based RKxxYxxY motif in immune cell adaptor SKAP55.

Src-homology 3 (SH3) domains recognize PXXP core motif preceded or followed by positively charged residue(s). Whether SH3 domains recognize motifs other than proline-based sequences is unclear. In this study, we report SH3 domain binding to a novel proline-independent motif in immune cell adaptor SKAP55, which is comprised of two N-terminal lysine and arginine residues followed by two tyrosines (i.e. RKxxYxxY). Domains capable of binding to class I proline motifs bound to the motif, while the class II domains failed to bind. Peptide precipitation, alanine scanning and in vivo co-expression studies demonstrated a requirement for the arginine, lysine and tandem tyrosines of the motif. Two-dimensional NMR analysis of the peptide bound FYN-SH3 domain showed overlap with the binding site of a proline-rich peptide on the charged surface of the SH3 domain, while resonance signals for other residues (W119, W120, Y137) were not perturbed by the RKGDYASY based peptide. Expression of the RKGDYASY peptide potently inhibited TcRzeta/CD3-mediated NF-AT transcription in T cells. Our findings extend the repertoire of SH3 domain binding motifs to include a tyrosine-based motif and demonstrate a regulatory role for this motif in receptor signaling.

Amino Acids, Diamino↗
PubMed2000

Utilization of site-directed spin labeling and high-resolution heteronuclear nuclear magnetic resonance for global fold determination of large proteins with limited nuclear overhauser effect data.

To test whether distances derived from paramagnetic broadening of (15)N heteronuclear single quantum coherence (HSQC) resonances could be used to determine the global fold of a large, perdeuterated protein, we used site-directed spin-labeling of 5 amino acids on the surface of (15)N-labeled eukaryotic translation initiation factor 4E (eIF4E). eIF4E is a 25 kDa translation initiation protein, whose solution structure was previously solved in a 3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate hydrate (CHAPS) micelle of total molecular mass approximately 45-50 kDa. Distance-dependent line broadening consistent with the three-dimensional structure of eIF4E was observed for all spin-label substitutions. The paramagnetic broadening effects (PBEs) were converted into distances for modeling by a simple method comparing peak heights in (15)N-HSQC spectra before and after reduction of the nitroxide spin label with ascorbic acid. The PBEs, in combination with HN-HN nuclear Overhauser effects (NOEs) and chemical shift index (CSI) angle restraints, correctly determined the global fold of eIF4E with a backbone precision of 2.3 A (1.7 A for secondary structure elements). The global fold was not correctly determined with the HN-HN NOEs and CSI angles alone. The combination of PBEs with simulated restraints from another nuclear magnetic resonance (NMR) method for global fold determination of large proteins (methyl-protonated, highly deuterated samples) improved the quality of calculated structures. In addition, the combination of the two methods simulated from a crystal structure of an all alpha-helical protein (40 kDa farnesyl diphoshphate synthase) correctly determined the global fold where neither method individually was successful. These results show the potential feasibility of obtaining medium-resolution structures for proteins in the 40-100 kDa range via NMR.

Amino Acid Sequence↗
PubMed2000

A space-time structure determination of human CD2 reveals the CD58-binding mode.

We describe a procedure for a space-time description of protein structures. The method is capable of determining populations of conformational substates, and amplitudes and directions of internal protein motions. This is achieved by fitting static and dynamic NMR data. The approach is based on the jumping-among-minima concept. First, a wide conformational space compatible with structural NMR data is sampled to find a large set of substates. Subsequently, intrasubstate motions are sampled by using molecular dynamics calculations with force field energy terms. Next, the populations of substates are fitted to NMR relaxation data. By diagonalizing a second moment matrix, directions and amplitudes of motions are identified. The method was applied to the adhesion domain of human CD2. We found that very few substates can account for most of the experimental data. Furthermore, only two types of collective motions have high amplitudes. They represent transitions between a concave (closed) and flat (open) binding face and resemble the change upon counter-receptor (CD58) binding.

Binding Sites↗
PubMed2000

Hox cluster genomics in the horn shark, Heterodontus francisci.

Reconstructing the evolutionary history of Hox cluster origins will lead to insights into the developmental and evolutionary significance of Hox gene clusters in vertebrate phylogeny and to their role in the origins of various vertebrate body plans. We have isolated two Hox clusters from the horn shark, Heterodontus francisci. These have been sequenced and compared with one another and with other chordate Hox clusters. The results show that one of the horn shark clusters (HoxM) is orthologous to the mammalian HoxA cluster and shows a structural similarity to the amphioxus cluster, whereas the other shark cluster (HoxN) is orthologous to the mammalian HoxD cluster based on cluster organization and a comparison with noncoding and Hox gene-coding sequences. The persistence of an identifiable HoxA cluster over an 800-million-year divergence time demonstrates that the Hox gene clusters are highly integrated and structured genetic entities. The data presented herein identify many noncoding sequence motifs conserved over 800 million years that may function as genetic control motifs essential to the developmental process.

Animals↗
PubMed2000

Iminoacylation. 3. Formation of platinum(IV)-based metallaligands due to facile one-end addition of vic-dioximes to coordinated organonitriles.

The reaction of vic-dioximes with the organonitrile platinum(IV) complexes trans-[PtCl4(RCN)2] (R = Me, CH2Ph, Ph, vic-dioxime = dimethylglyoxime; R = Me, vic-dioxime = cyclohexa-, cyclohepta-, and cyclooctanedione dioximes) proceeds rapidly under relatively mild conditions and affords products of one-end addition of the dioximes to the nitrile carbon, i.e. [PtC4(NH=C(R)ON=[spacer]=NOH)2] (1-6) (R = Me, CH2Ph, Ph, spacer = C(Me)C-(Me) for dimethylglyoxime; R = Me, spacer = C[C4H8]C, C[C5H10]C, C[C6H12]C for the other dioximes), giving a novel type of metallaligand. All addition compounds were characterized by elemental analyses (C, H, N, C1, Pt), FAB mass spectrometry, and IR and 1H, 13C[1H], and 195Pt NMR spectroscopy. X-ray structure determination of the dimethylformamide bis-solvate [PtCl4(NH=C(Me)ON=C(Me)C(Me)=NOH)2] x 2DMF (la) disclosed its overall trans geometry with the dimethylglyoxime part in anti configuration and the amidine one-end (rather than N,N-bidentate) coordination mode of the N-donor ligands. When a mixture of cis- and trans-[PtC4(MeCN)2] in MeCN was treated with dimethylglyoxime, the formation of, correspondingly, cis- and trans-[PtCl4(NH=C(Me)ON=C(Me)C(Me)=NOH)2] (1) was observed and cis-to-trans isomerization in DMSO-d6 solution was monitored by 1H, 2D [1H,15N] HMQC, and 195Pt NMR spectroscopies. Although performed ab initio calculations give evidence that the trans geometry is the favorable one for the iminoacylated species [PtCl4-(ligand)2], the platinum(IV) complex [PtCl4(NH=C(Me)ON=C[C4Hs]C=NOH)2] (4) was isolated exclusively in cis configuration with the two metallaligand "arms" held together by intramolecular hydrogen bonding between the two peripheral OH groups, as it was proved by single-crystal X-ray diffractometry. The classic substitution products, e.g. [PtC12(N,N-dioximato)2] (12-15), are formed in the addition reaction as only byproducts in minor yield; two of them, [PtCl2(C7H11N2O2)2] (14) and [PtCl2(C8H13N2O2)2] (15), were structurally characterized. Complexes (12-15) were also prepared by reaction of the vic-dioximes with [PtCl4L(Me2SO)] (L = Me2SO, MeCN), but monoximes (Me2C=NOH, [C4H8]C=NOH, [C5H10]C=NOH, PhC(H)=NOH, (OH)C6H4C(H)= NOH) react differently adding to [PtCl4(MeCN)(Me2SO)] to give the corresponding iminoacylated products [PtCl4(NH=C(Me)ON=CRR')(Me2SO)](7-11).

Journal Article↗
PubMed2000

Canalization in evolutionary genetics: a stabilizing theory?

Canalization is an elusive concept. The notion that biological systems ought to evolve to a state of higher stability against mutational and environmental perturbations seems simple enough, but has been exceedingly difficult to prove. Part of the problem has been the lack of a definition of canalization that incorporates an evolutionary genetic perspective and provides a framework for both mathematical and empirical study. After briefly reviewing the importance of canalization in studies of evolution and development, we aim, with this essay, to outline a research program that builds upon the definition of canalization as the reduction in variability of a trait, and uses molecular genetic approaches to shed light on the problems of canalization.

Adaptation, Physiological↗