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Biomedical subjects

M Schauer

Publications and source records attributed to M Schauer.

At least 19 recordsLinked to original sources

Spatial differences in bacterioplankton composition along the Catalan coast (NW Mediterranean) assessed by molecular fingerprinting.

Denaturing gradient gel electrophoresis (DGGE) of PCR-amplified 16S rRNA gene fragments was used to compare surface bacterioplankton assemblages along the Catalan coast (NW Mediterranean). Samples from three coastal stations were compared with samples taken inside the Barcelona harbour and open sea samples taken during a cruise. The bacterial assemblage of each sample showed a characteristic and reproducible DGGE fingerprint. Between 17 and 35 bands were detected in each sample, and about 40% of the bands accounted for more than 80% of the band intensity in each sample. The presence of bands as well as their relative intensity was used to compare bacterial assemblages. Clear differences between the harbour samples and the coastal samples were evident during all periods. Marked temporal changes in the bacterial assemblages were detectable for the coastal sites, suggesting seasonal succession of coastal bacterioplankton. During each season, two stations presented a very similar bacterial composition (Barcelona and Masnou) whereas bacterial assemblages in Blanes were slightly different. These differences were consistent with the different hydrography of the area. Diversity indices calculated from DGGE fingerprints were relatively similar for all samples analysed, even though harbour samples were expected to present lower diversity values.

Journal Article↗

Two assays for measuring fibrosis: reverse transcriptase-polymerase chain reaction of collagen alpha(1) (III) mRNA is an early predictor of subsequent collagen deposition while a novel serum N-terminal procollagen (III) propeptide assay reflects manifest fibrosis in carbon tetrachloride-treated rats.

Using a novel quantitative reverse transcriptase-polymerase chain reaction assay, we have determined the amount of specific mRNA for procollagen alpha(1) (III) (PIIIP) in the carbon tetrachloride (CCl(4)) model of liver fibrosis in rats. After a single week of CCl(4) application, the amount of PIIIP mRNA was increased approximately 10 times over the untreated control group and continued to increase to approximately 30 times after 7 weeks of intoxication. In this model substantial fibrosis was demonstrated by computer-aided morphometry after 5 to 7 weeks of treatment. Using recombinant murine N-terminal procollagen alpha(1) (III) propeptide (PIIINP), a novel sensitive immunoassay for the measurement of circulating PIIINP in rodent sera was established. An increase in PIIINP serum levels was observed after 5 to 7 weeks of CCl(4) intoxication. Our results suggest PIIIP gene expression is an early marker of tissue fibrosis. Early PIIIP gene expression is correlated with the extent of the subsequent fibrosis. PIIIP mRNA levels increase much earlier than conventional histological examination or PIIINP levels. PIIINP measurements with our new serum assay, on the other hand, are a good noninvasive marker of manifest fibrosis but are a poor marker of fibrogenesis.

Animals↗

The thought translation device: a neurophysiological approach to communication in total motor paralysis.

A thought translation device (TTD) for brain-computer communication is described. Three patients diagnosed with amyotrophic lateral sclerosis (ALS), with total motor paralysis, were trained for several months. In order to enable such patients to communicate without any motor activity, a technique was developed where subjects learn to control their slow cortical potentials (SCP) in a 2-s rhythm, producing either cortical negativity or positivity according to the task requirement. SCP differences between a baseline interval and an active control interval are transformed into vertical or horizontal cursor movements on a computer screen. Learning SCP self regulation followed an operant-conditioning paradigm with individualized shaping procedures. After prolonged training over more than 100 sessions, all patients achieved self-control, leading to a 70-80% accuracy for two patients. The learned cortical skill enabled the patients to select letters or words in a language-supporting program (LSP) developed for inter-personal communication. The results demonstrate that the fast and stable SCP self-control can be achieved with operant training and without mediation of any muscle activity. The acquired skill allows communication even in total locked-in states.

Adult↗

Cultured epidermal keratinocytes on a microspherical transport system are feasible to reconstitute the epidermis in full-thickness wounds.

Research efforts to modify cultured autologous skin transplants for large full-thickness burn wounds and in chronic ulcers have shifted from multilayered differentiated grafts ("sheet" grafts) toward smaller units of basal undifferentiated single cell suspensions in a transport medium and subconfluently covered static carriers. It has been shown that wounds transplanted with single cell suspensions reconstitute the epidermis. However, this technique requires the detachment of the keratinocytes from the culture flasks by enzymatic digestion-digestion that might alter the anchoring proteins of the cells. A new approach might be to circumvent the enzymatic digestion to harvest the keratinocytes. This study reports a technique to culture epidermal cells on spherical microcarriers as a suspension culture and transport vehicle. The spherical microcarrier consists of a 100-microm-diameter collagen-coated dextran carrier (Cytodex 3 Pharmacia) and has been used previously for enzyme production commercially. With this new approach, we seeded the human keratinocytes in a spinner-like system onto microspheres and transplanted these micrografts onto full-thickness wounds on the back of nude mice. After 14 days, we showed a reconstituted epithelium that was multilayered and keratinized compared to control wounds. We believe that this is the first step of a new approach to increase the cell yield for seeding without altering the anchoring proteins by enzymatic steps, leading to a superior transplantation method for keratinocytes.

Adult↗

Epitope-specific monoclonal antibodies against human C-terminal procollagen alpha1(III)-propeptide.

We have generated monoclonal antibodies against recombinant C-terminal human procollagen alpha1(III) propeptide (PIIICP), produced in E. coli in high yields. The monoclonal antibodies were screened for epitope specificity using recombinant truncated PIIICP. Several antibodies were identified which recognized different regions of the PIIICP molecule. The ability of the antibodies to detect PIIICP antigens in human cell line lysates and supernatants was demonstrated. As PIIICP antigens are a key marker of extracellular matrix metabolism, the monoclonal antibodies described here should be of value for clinical and basic research.

Antibodies, Monoclonal↗

Sit-to-stand manoeuvre in hemiparetic patients before and after a 4-week rehabilitation programme.

The objective of the study was to document the sit-to-stand of 35 moderately affected hemiparetic patients before and after a 4-week inpatient rehabilitation programme and to investigate the influence of the side of hemiparesis and impaired proprioception on its outcome. The therapists trained the patients to distribute equal weight on both legs and to avoid lateral compensatory tilt of the trunk. The body weight distribution and the displacement of the centre of mass in the frontal plane were studied by assessment of triaxial ground reaction forces. Symmetrical weight acceptance and minimal lateral sway were not reached during the 4-week rehabilitation, with the exception of the patients with a left hemiparesis (n = 19) and no proprioceptive deficit (n = 18).

Aged↗

[Effect of music on gait symmetry of stroke patients on a treadmill].

The influence of rhythmic music on gait symmetry was investigated in 12 healthy subjects and 12 stroke patients with mild leg paresis walking on the treadmill. For the measurement, new insoles containing air-filled chambers developed by W.O.M were used. Symmetry deviation was determined as the mean signless difference between left and right swing phases of some 100 strides. In 6 patients, the symmetry deviation decreased by more than 1% of the stride duration some 40 steps after switching on the music. The improvement correlated with the initial symmetry (r2 = 0.61) exclusively in the healthy controls. In order to differentiate individual predictors of the improvement in symmetry, such as cognitive performance in terms of recognizing the beat of the music and the motor performance in terms of synchronizing the movements of the legs to the music, we analysed foot tapping with the patient in the seated position. For both patients and healthy subjects, the mean time difference between beat and foot movement was less than +/-1/32 beat. Individual scatter of mean 8% correspond in the case of healthy subjects to the figures found in the literature for finger movements, and the corresponding figures for the patients are more than twice this. The individual synchronisation performance during walking to music correlates to that during foot tapping (r2 = 0.47) exclusively in the group of healthy controls.

Cerebrovascular Disorders↗

Bacteria- and endotoxin-free dialysis fluid for use in chronic hemodialysis.

As the quality of water in the dialysis fluid varies considerably, dialysis fluid is contaminated with a high percentage of bacteria and endotoxins. The bacterial populations contained in the dialysis fluid are as heterogeneous as the chemical structure of the endotoxins that result. The latter can pass through the dialysis membrane whereby high-flux membranes permit a larger number of retransportable molecules than low-flux membranes. A central aim toward a future, safe dialysis process should, therefore, be the production of a dialysate that is free of bacteria and endotoxins. As we were able to demonstrate in various examinations, this goal is most likely to be achieved with the aid of sterile filtration using hollow fiber modules of polyamid. To avoid disinfection of the polyamid membrane, as this would only reach bacteria but not endotoxins, the filter was changed after at most 10 h. The achieved dialysis fluid was free of bacteria and endotoxins. We were also able to show that the release of interleukin-1 was reduced. In addition, side-effects, such as a drop in blood pressure, headaches, muscular cramps, and nausea, were reduced.

Bacteria↗

Quantitative analysis of rising from a chair in healthy and hemiparetic subjects.

In 15 healthy and 20 hemiparetic persons we studied standing-up by two force-plates. Phases before and after the seat-off, force distribution and centre of gravity displacement were assessed. The patients rose significantly slower. The force ratios after seat-off differed significantly between the groups (0.88 vs 0.68). Left/right hemiparetic patients put more weight on the affected limb in 18%/11% of the trials before seat-off, with its insufficient use after it mainly in the left patients. At seat-off, projection of the centre of gravity fell within the support area in hemiparetic patients, and 3 cm behind it in healthy subjects. With larger lateral displacement of the centre of gravity in the hemiparetic group, left hemiparetic patients mostly shifted their weight to the non-affected side and right hemiparetic patients to both sides. Weight distribution and mediolateral displacement of the centre of gravity in the left and right hemiparetic patients were considered.

Activities of Daily Living↗

[A clinically applicable system for analysis of standing up using measurements of floor reaction forces].

A system for clinical analysis of sit-to-stand transfer using two force plates is presented. Distinct time-events and phases are estimated. Illustrative parameters are also determined, such as the distribution of the vertical ground reaction forces, displacement of the center of gravity in the sagittal and frontal planes, and a comparison of the displacement of the center of gravity and the center of pressure. These parameters indicate a symmetrical raising while keeping the middle of the body. Future applications include the validation of therapy, biofeedback and the control of closed-loop FES.

Biomechanical Phenomena↗

The N-terminal region of HIV-1 integrase is required for integration activity, but not for DNA-binding.

HIV-1 integrase binds to both double- and single-stranded DNA with Kd-values of around 20 nM, irrespective of sequence similarities with the termini of the viral LTR. For integration activity, however, the correct LTR sequence of the substrate is required. The putative zinc-binding site present at the N-terminus of the protein is not essential for DNA binding, since deletion mutants of the protein lacking this sequence show similar affinity towards DNA as the wild-type; however, these mutants are not capable of performing the LTR-cleavage and integration reactions. Thus, it appears that the N-terminal part of the integrase is essential for catalytic activity.

Base Sequence↗

Efficient binding of glucocorticoid receptor to its responsive element requires a dimer and DNA flanking sequences.

A combination of the gel retardation assay and interference by hydroxyl radical modification (missing nucleoside technique) was used to analyze the interaction of the glucocorticoid receptor (GR) with various glucocorticoid responsive elements (GRE). Short oligonucleotides containing the 15-bp GRE and 1 to 3 flanking base pairs on each side, are bound with very low affinity. The same GREs, when positioned in the center of a large DNA fragment (40-50 bp), show high affinity for the receptor. However, when the GRE is positioned at the border of a 54-bp fragment, the affinity of the GR for the GRE decreases markedly. The DNA binding affinity increases linearly with each added flanking base pair and optimal binding is observed with 8-10 flanking bp. Thus, the nonconserved DNA sequences flanking the GRE contribute significantly to the free energy of receptor binding to DNA. Using larger DNA fragments (greater than 100 bp) and a smaller form of the receptor (40 kD), two retarded complexes are found that correspond to monomeric and homodimeric receptor DNA complexes. The DNA-binding domain of the GR (20 kD), expressed in bacteria, binds to the GRE as a monomer as well as a dimer and can form heterodimers with the native 94-kD GR. Insertion or deletion of one single base pair between the two halves of the GRE reduces the affinity for the homodimeric form of the native GR, and inhibits the function of the GRE in gene transfer experiments, suggesting that a dimer of the GR is the functional entity that binds to the GRE.

Animals↗

DNA regulatory elements for steroid hormones.

Gene regulation by steroid hormones is mediated through an interaction of the hormone receptors with DNA regulatory sequences called hormone regulatory or responsive elements (HRE). An analysis of the HRE's in the DNA of mouse mammary tumour provirus, human metallothionein IIA gene, chicken lysozyme gene, chicken and Xenopus vitellogenin genes, growth hormones genes, Moloney murine sarcoma provirus, rabbit uteroglobin gene, rat tyrosine aminotransferase gene, rat tryptophan oxygenase gene and rat acidic glycoprotein gene, yields the following consensus for positively modulated glucocorticoid responsive elements (GRE): 5'-GGTACAnnnTGTTCT-3'. This element can also mediate induction by progesterone and probably by androgens, but not by estrogens. Detailed analysis of the DNA protection pattern suggests that a dimer of the hormone receptor interacts with this palindromic 15-mer. In genes that are negatively regulated by glucocorticoids an imperfect copy of the GRE is found, and repression is probably due to competition between hormone receptor and other transcription factors or enhancer binding proteins for binding to overlapping DNA sequences. The receptors without bound hormone are able to interact specifically with DNA in vitro, but binding of hormone is needed for transcriptional activation in vivo. This could be due, at least in part, to changes in the rate parameters of the receptor-DNA interaction induced by binding of the hormone to the receptor. The possible role of precise chromatin organization in glucocorticoid induction is discussed on the basis of the nucleosome phasing found in the LTR region of mouse mammary tumour virus.

Animals↗

Binding of hormone accelerates the kinetics of glucocorticoid and progesterone receptor binding to DNA.

Steroid hormone receptors induce genes by virtue of their interaction with DNA regulatory sequences. The hormonal response of a particular gene in vivo correlates with binding of the hormone to the receptor and supposedly reflects the degree of occupancy of the corresponding DNA regulatory sequences. However, in vitro the steroid-free glucocorticoid and progesterone receptors bind specifically to the regulatory sequences of mouse mammary tumor virus, thus raising questions on the role of the hormone in DNA binding in vivo. By using monoclonal antibodies, gel retardation assays, and filter binding techniques we show here that binding of a functional steroid to either the glucocorticoid or the progesterone receptors influences the kinetics of the protein-DNA interaction in vitro. In the presence of hormone the on rate of receptor binding to DNA fragments with or without regulatory sequences is accelerated 2- to 5-fold, and the off rate is accelerated 10- to 20-fold. The receptors complexed to an antihormone bind to DNA with kinetics intermediate between those of the steroid-free and the hormone-bound protein. Thus, ligand binding accelerates the kinetics of receptor binding to DNA and this could partly account for the behavior of the hormone receptor observed in vivo.

Adrenalectomy↗

[Mathematical modelling of glycolysis and adenine nucleotide metabolism of human erythrocytes. I. Reaction-kinetic statements, analysis of in vivo state and determination of starting conditions for in vitro experiments].

A mathematical model of energy metabolism of human red cells is presented, which includes besides the glycolytic reactions the adenine nucleotide metabolism. The model is based on the network of chemical reactions, the thermodynamic equilibrium constants of fast reversible reactions and on the kinetic equations for irreversible enzyme reactions. The model consists of a system of 16 differential equations and allows the mathematical evaluation of metabolic levels in the steady state of energy metabolism corresponding to the in vivo state erythrocytes with the kinetic data for the enzymes derived from in vitro experiments. The dependence of the levels of metabolites in the steady state on the activity of some enzymes is analysed to characterize the regulatory properties of the system. The comparison of the steady state levels of the model with experimental data makes it possible to estimate values of some controversial enzyme parameters. Estimates of the kinetic parameters of the following intracellular processes are presented: 1) rate constant of AMP-phosphatase, 2) maximum rate of adenylate deaminase, 3) activity of adenine phosphoribosylpyrophosphate transferase and 4) adenosine transport through the cell membrane. The simulation of the preparatory phase before incubation of erythrocytes indicates, that the model also permits to compute the time course of changes of levels of metabolites. To solve the initial problem the stiff differential equation system is integrated numerically by an efficient program without the application of the quasi-steady-state approximation.

Adenine Nucleotides↗