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Biomedical subjects

S Müller

Publications and source records attributed to S Müller.

At least 73 records · Page 4Linked to original sources

Recruitment of spinal motor pools during voluntary movements versus stepping after human spinal cord injury.

We investigated the activation of lower limb motor pools by supraspinal and spinal networks after human spinal cord injury (SCI). We compared electromyographic (EMG) activity from six muscles during voluntarily attempted non-weight-bearing single-joint movements, multijoint movements approximating stepping in a supine position, and weight-bearing stepping on a treadmill with body weight support (BWST) in seven clinically incomplete and three clinically complete SCI subjects. Seven SCI subjects had previously completed Laufband therapy (a specific step training using variable levels of body weight support and manual assistance). Significant coactivation of agonists and antagonists and multijoint flexion or extension movements of the entire limb occurred during attempts at isolated knee or ankle single-joint movements in clinically incomplete SCI subjects. Further, some muscles that were not recruited during voluntary attempts at single-joint movements were activated during voluntary step-like multijoint movements (5/16 comparisons). This suggests that the residual voluntary motor control in incomplete SCI subjects evokes more generalized motor patterns (limb flexion or extension) rather than selective activation of individual muscles. Clinically incomplete and clinically complete SCI subjects could achieve greater activation of motor pools and more reciprocal patterns of activity between agonists and antagonists during weight bearing stepping than during non-weight-bearing voluntary movements. The EMG mean amplitudes were higher during stepping than during voluntary movements in 50/60 muscles studied (p < 0.05). These results suggest that stepping with knee and hip extension and flexion and alternating lower limb loading and unloading provides proprioceptive inputs to the spinal cord that increases motor recruitment and improves reciprocity between agonists and antagonists compared to voluntary efforts.

Adolescent↗

[Biventricular defibrillation using transvenous electrodes].

This study investigated the feasibility of transvenous biventricular defibrillation using an electrode in a left ventricular vein. The standard lead configuration with a coil in the right ventricle (RV) and a coil in the superior vena cava (SVC) was compared with an additional unipolar coil in an accessible epicardial vein. Only biphasic shocks were used with different shocking modes between the coils in the RV, LV, SVC and the ICD-generator (CAN). Shocks were applied starting with 30 J, decreasing till the DFT was reached. As a result there is a lower DFT when defibrillation is performed including the left ventricular electrode. The impedance did not show a significant increase after more then 20 consecutive shocks. It is a feasible and workable application that might help to reduce the energy demand and increase the safety of such a system.

Animals↗

[Blood gas regulation in isolated beating pig hearts].

Continuous perfusion with oxygenated blood is required to supply isolated beating pig hearts. To meet physiological conditions the blood gas parameters pH, pO2 and pCO2 should correspond with values usually found in pigs, respectively. Controlling these parameters manually is a very time consuming task and is therefore done best by a control system consisting of gas blender, oxygenator and blood gas sensors. As this control system should be used with an transportable perfusion apparatus, this paper describes an approach where the gassing is realized only with air and oxygen. In this approach pO2 and pH are measured on blood side so that the control system can change oxygen concentration and gas flow adequately on the gas side.

Acid-Base Equilibrium↗

Image-guided navigation for minimal invasive approaches in craniomaxillofacial surgery.

The use of minimally invasive procedures in maxillofacial surgery will require new technologies involving surgical navigation and techniques. The aim of our studies is to improve the efficacy of image-guided navigation in combination with endoscopically assisted techniques for minimally invasive craniomaxillofacial procedures. Prospective evaluation was made of all patients who underwent surgical procedures using image-guided navigation. The most common type of operations performed were endoscopically assisted interventions within the paranasal sinuses, fracture treatment, the resection of bone lesions and further miscellaneous interventions. Our experience to date suggest that image-data based techniques are eminently applicable, providing a feasible alternative to conventional surgical treatment.

Endoscopes↗

[Fiberoptic measurement of myocardial contraction--correlation of the signal with hemodynamic values].

In addition to the intracardial ECG, the mechanical myocard contraction should be used as another input signal for cardiac pacemakers and implantable defibrillators. Therefore a fiberoptical measurement system was designed. A sensor-fiber was placed in the coronary venous system. An opto-electronic system converts the optical losses, caused by bending of the fiber, into a proportional voltage. This method allows measuring of the left ventricular myocard contraction strength. By theoretical calculations it was shown, that a good correlation of the sensor-signal and the left ventricular radius can be expected. Additional investigations using an isolated beating pig heart were performed. A high correlation of the sensor-signal and the left ventricular stroke volume was shown.

Animals↗

First-principles predictions of yet-unobserved ordered structures in the Ag-Pd phase diagram.

The complexity of first-principles total energy calculations limits the pool of structure types considered for a ground-state search for a binary alloy system to a rather small, O(10), group of "usual suspects." We conducted an unbiased search of fcc-based Ag(1)-(x)Pd(x) structures consisting of up to many thousand atoms by using a mixed-space cluster expansion. We find an unsuspected ground state at 50%-50% composition-the L1(1) structure, currently known in binary metallurgy only for the Cu(0.5)Pt(0.5) alloy system. We also provide predicted short-range-order profiles and mixing enthalpies for the high temperature, disordered alloy.

Journal Article↗

Mammalian cells express two VPS4 proteins both of which are involved in intracellular protein trafficking.

The yeast Vps4 protein (Vps4p) is a member of the AAA protein family (ATPases associated with diverse cellular activities) and a key player in the transport of proteins out of a prevacuolar endosomal compartment. In human cells, we identified two non-allelic orthologous proteins (VPS4-A and VPS4-B) of yeast Vps4p. The human VPS4-A and VPS4-B proteins display a high degree of sequence identity to each other (80 %) and to the yeast Vps4 protein (59 and 60 %, respectively). Yeast cells lacking a functional VPS4 gene exhibit a temperature-sensitive growth defect and mislocalise a carboxypeptidase Y-invertase fusion protein to the cell surface. Heterologous expression of human VPS4 genes in vps4 mutant yeast strains led, in the case of human VPS4-A, to a partial and, in the case of human VPS4-B, to a complete suppression of the temperature-sensitive growth defect. The vacuolar protein sorting defect of vps4 mutant yeast cells was complemented completely by heterologous expressed human VPS4-B protein, and partially by the human VPS4-A protein. Expression of mutant human VPS4-A (E228Q) and VPS4-B (E235Q) proteins, harbouring single amino acid exchanges in their AAA domains, induced dominant-negative vacuolar protein sorting defects in wild-type yeast cells in both cases. Two-hybrid experiments suggest that the human VPS4-A and VPS4-B proteins can form heteromeric complexes, and subcellular localisation experiments indicate that both human VPS4 proteins associate with endosomal compartments in yeast. Based on these results, we conclude that both human VPS4 proteins are involved in intracellular protein trafficking, presumably at a late endosomal protein transport step, similar to the Vps4p in yeast.

ATPases Associated with Diverse Cellular Activitie↗

Detection of a conformational change in G gamma upon binding G beta in living cells.

Interaction induced changes in the conformation of proteins are frequently the molecular basis for the modulation of their activities. Although proteins perform their functions in cells, surrounded by many potential interaction partners, the studies of their conformational changes have been mainly restricted to in vitro studies. Ste4p (G beta) and Ste18p (G gamma) are the subunits of a heterotrimeric G-protein in the yeast Saccharomyces cerevisiae. A split-ubiquitin based conformational sensor was used to detect a major structural rearrangement in Ste18p upon binding to Ste4p. Based on these in vivo results and the solved structure of the mammalian G beta gamma, we propose that G gamma of yeast adopts an equally extended structure, which is only induced upon association with G beta.

Fungal Proteins↗

Thermodynamics of HMGB1 interaction with duplex DNA.

The high mobility group protein HMGB1 is a small, highly abundant protein that binds to DNA in a non-sequence-specific manner. HMGB1 consists of 2 DNA binding domains, the HMG boxes A and B, followed by a short basic region and a continuous stretch of 30 glutamate or aspartate residues. Isothermal titration calorimetry was used to characterize the binding of HMGB1 to the double-stranded model DNAs poly(dAdT).(dTdA) and poly(dGdC).(dCdG). To elucidate the contribution of the different structural motifs to DNA binding, calorimetric measurements were performed comparing the single boxes A and B, the two boxes plus or minus the basic sequence stretch (AB(bt) and AB), and the full-length HMGB1 protein. Thermodynamically, binding of HMGB1 and all truncated constructs to duplex DNA was characterized by a positive enthalpy change at 15 degrees C. From the slopes of the temperature dependence of the binding enthalpies, heat capacity changes of -0.129 +/- 0.02 and -0.105 +/- 0.05 kcal mol(-1) K(-1) were determined for box A and full-length HMGB1, respectively. Significant differences in the binding characteristics were observed using full-length HMGB1, suggesting an important role for the acid tail in modulating DNA binding. Moreover, full-length HMGB1 binds differently these two DNA templates: binding to poly(dAdT).(dTdA) was cooperative, had a larger apparent binding site size, and proceeded with a much larger unfavorable binding enthalpy than binding to poly(dGdC).(dCdG).

Aspartic Acid↗

Head-to-head comparison of N-terminal pro-brain natriuretic peptide, brain natriuretic peptide and N-terminal pro-atrial natriuretic peptide in diagnosing left ventricular dysfunction.

Brain natriuretic peptide (BNP), NT-proBNP and NT-pro-atrial natriuretic peptide (NT-proANP) were measured in blood samples from 57 patients using immunoassays and immunoradiometric assays to evaluate the usefulness as diagnostic markers for the detection of heart failure. For the detection of impaired left ventricular ejection fraction (LVEF), receiver operating characteristic curves showed that BNP had the best diagnostic performance with an area under curve (AUC) of 0.75+/-0.06. However, NT-proBNP (AUC: 0.67+/-0.07) and NT-proANP (AUC: 0.69+/-0.08) showed no significant difference to BNP. In a further analysis for the detection of resting LVEF <40%, BNP again was the best marker with an AUC of 0.83+/-0.06. NT-proBNP showed only a slightly smaller AUC (0.79+/-0.07). The AUC for NT-proANP was significantly smaller (0.65+/-0.08) compared to BNP. Additionally, BNP and NT-proBNP correlated negatively with the resting LVEF (BNP: -0.472, p<0.001; NT-proBNP: -0.306, p=0.026), whereas NT-proANP showed no significant correlation. In summary, BNP was the best marker to detect patients with impaired LVEF compared to NT-proBNP and NT-proANP. However, NT-proBNP showed no significant differences to BNP and it is therefore a new promising alternative marker for the detection of left ventricular dysfunction.

Aged↗

Astrocytes induce manganese superoxide dismutase in brain capillary endothelial cells.

Astrocytes induce blood-brain barrier (BBB) properties in brain endothelial cells (EC)*O(2)*, generated in blood and EC, opens the BBB. Hence, high activity of superoxide dismutase (SOD) is a prerequisite for normal BBB function. Therefore, the influence of rat astrocytes on the expression of manganese (Mn)SOD in rat EC was investigated in two coculture models of the BBB, allowing either exchange of soluble factors or additionally cellular contacts. Activity, protein content and mRNA expression of endothelial MnSOD were significantly increased in both coculture models in comparison to monoculture by soluble astrocytic factors, such as cytokines. High activity of endothelial MnSOD may be considered as a further essential property of the BBB, which is induced and maintained by astrocytes.

Animals↗

Structural characterization of human recombinant and bone-derived bone sialoprotein. Functional implications for cell attachment and hydroxyapatite binding.

Human bone sialoprotein (BSP) comprises 15% of the total noncollagenous proteins in bone and is thought to be involved in bone mineralization and remodeling. Recent data suggest a role for BSP in breast cancer and the development of bone metastases. We have produced full-length recombinant BSP in a human cell line and purified the protein from human bone retaining the native structure with proper folding and post-translational modifications. Mass spectrometry of bone-derived BSP revealed an average mass of 49 kDa and for recombinant BSP 57 kDa. The post-translational modifications contribute 30-40%. Carbohydrate analysis revealed 10 different complex-type N-glycans on both proteins and eight different O-glycans on recombinant BSP, four of those were found on bone-derived BSP. We could identify eight threonines modified by O-glycans, leaving the C terminus of the protein free of glycans. The recombinant protein showed similar secondary structures as bone-derived BSP. BSP was visualized in electron microscopy as a globule linked to a thread-like structure. The affinity for hydroxyapatite was higher for bone-derived BSP than for recombinant BSP. Cell adhesion assays showed that the binding of BSP to cells can be reversibly diminished by denaturation.

Amino Acid Sequence↗

Cloning and expression of cDNAs encoding alpha1,3-fucosyltransferase homologues from Arabidopsis thaliana.

The core alpha1,3-fucosyltransferases are involved in the synthesis of glycans specific to plants and invertebrates which are known to be immunogenic and allergenic. We report the identification, isolation and characterisation of the cDNAs of three genes (FucTA, FucTB and FucTC) encoding proteins similar to alpha1,3-fucosyltransferases in Arabidopsis thaliana. Reverse transcription-polymerase chain reaction was used to amplify the full length coding sequence of FucTA. The FucTA gene, which consists of seven exons, encodes a presumptive protein of 501 amino acids showing an overall sequence identity of 66% to the protein encoded by the recently isolated mung bean Fuc-T C3 cDNA. FucTA was expressed in Pichia pastoris under the control of the AOX1 gene promoter. The soluble enzyme was found to catalyse the same reaction as mung bean core alpha1,3-fucosyltransferase as judged by analyses of the products by MALDI-TOF and high-performance liquid chromatography. The FucTB cDNA was isolated from a lambda-ZAP library, but the clone used an alternative splicing site between the second and third exon resulting in a premature stop codon. The FucTC gene encodes a protein with less than 40% identity to FucTA across 115 amino acids of a total of 401 amino acids and is a member of a new sub-family of plant alpha1,3/4-fucosyltransferase homologues.

Amino Acid Sequence↗

Low prevalence of latently Epstein-Barr virus-infected cells in chronic gastritis.

The association of Epstein-Barr virus (EBV) with a proportion of gastric carcinomas is well established. The role of EBV in conditions predisposing to carcinoma such as chronic gastritis has remained undefined, however. We used in situ hybridization with radioactive and nonradioactive single-stranded RNA probes specific for the EBV small latent nuclear transcripts, EBER1 and EBER2, to analyze biopsy specimens from 242 patients with mild to severe chronic gastritis of Sydney classification types A, B, and C. A small number of EBV infected lymphocytes was detected in only nine cases, even in biopsies investigated with radioactive probes. Labeling of epithelial or stromal cells was not observed. The paucity of latently EBV-infected cells in chronic gastritis biopsies differs from the previously reported higher prevalence of virus carrying cells in inflammatory conditions at other sites of the gastrointestinal tract. These findings argue against a direct involvement of EBV in the pathogenesis of chronic gastritis. The low prevalence of EBV-positive cells suggests that local factors do not favor the entry and retention of circulating EBV-infected lymphocytes in gastric mucosa. Moreover, our findings indicate that EBV infection of gastric epithelial cells is not an early event in gastric carcinogenesis.

Adolescent↗

Spermine supports catalysis of hairpin ribozyme variants to differing extents.

Because of the ability to cleave RNA substrates in trans, the hairpin ribozyme has great potential for therapeutic application. Activity of a three-stranded version of the minimal truncated form is enhanced by the presence of the polyamine spermine. Since spermine is the most abundant polyamine in eucariots, improved prospects for the hairpin ribozyme as therapeutic agent were predicted. We have found that not all hairpin ribozyme variants accept spermine equally well as counter-ion. Particularly the two-stranded versions commonly used for therapeutic studies show rather decreased activity when spermine is present. We have investigated a number of hairpin ribozyme derivatives regarding their ability to carry out spermine supported catalysis. Among the studied structures a two-stranded reverse-joined hairpin ribozyme displayed the highest cleavage rates in a synergistic mixture of magnesium ions and spermine. The specific features of this ribozyme along with its potential for in vivo application are discussed.

Base Sequence↗

The high mobility group (HMG) boxes of the nuclear protein HMG1 induce chemotaxis and cytoskeleton reorganization in rat smooth muscle cells.

HMG1 (high mobility group 1) is a ubiquitous and abundant chromatin component. However, HMG1 can be secreted by activated macrophages and monocytes, and can act as a mediator of inflammation and endotoxic lethality. Here we document a role of extracellular HMG1 in cell migration. HMG1 (and its individual DNA-binding domains) stimulated migration of rat smooth muscle cells in chemotaxis, chemokinesis, and wound healing assays. HMG1 induced rapid and transient changes of cell shape, and actin cytoskeleton reorganization leading to an elongated polarized morphology typical of motile cells. These effects were inhibited by antibodies directed against the receptor of advanced glycation endproducts, indicating that the receptor of advanced glycation endproducts is the receptor mediating the HMG1-dependent migratory responses. Pertussis toxin and the mitogen-activated protein kinase kinase inhibitor PD98059 also blocked HMG1-induced rat smooth muscle cell migration, suggesting that a G(i/o) protein and mitogen-activated protein kinases are required for the HMG1 signaling pathway. We also show that HMG1 can be released by damage or necrosis of a variety of cell types, including endothelial cells. Thus, HMG1 has all the hallmarks of a molecule that can promote atherosclerosis and restenosis after vascular damage.

Animals↗