Biomedical subjects

R Wirth

Publications and source records attributed to R Wirth.

At least 37 records · Page 2Linked to original sources

PubMed1992

Efficacy of different irrigation methods and concentrations of root canal irrigation solutions on bacteria in the root canal.

The effectiveness of two different root canal irrigating solutions, each in two different concentrations or formulations, with two different irrigation methods was compared in vitro by means of bacterial survival determinations. 75 human root canals were enlarged, sterilized and inoculated with a mixed culture of Escherichia coli and Streptococcus mutans. After inoculation, the root canals were irrigated either manually or with an ultrasonic device for equal times (20s) with the same amount (5 ml) of sodium hypochlorite (1% and 2%), Fokalhydran I and Fokalhydran II. Sodium hypochlorite (1% and 2%) was used in a 1:100 dilution. Fokalhydran I and II were used in a 1:10 dilution. In the sodium hypochlorite group, the 1% concentration applied with a syringe proved to be most effective against Escherichia coli and Streptococcus mutans. The least effective concentration and application method against both bacteria species was obtained with 2% NaOCL and ultrasonics. Against Escherichia coli and Streptococcus mutans, a significantly lower effectiveness was found with 2% NaOCL applied with ultrasonics with respect to the rest of the sodium hypochlorite group. Fokalhydran I was significantly better than Fokalhydran II against Escherichia coli. However, no significant differences could be seen against Streptococcus mutans within this group.

Analysis of Variance↗

PubMed1992

Aggregation substance of Enterococcus faecalis mediates adhesion to cultured renal tubular cells.

The sex pheromone system of Enterococcus faecalis is a unique, highly efficient plasmid collection mechanism for this species. A crucial role in this system is played by an adhesin called aggregation substance which enables the cell-cell contact between donor and recipient strains. The existence of the amino acid motif Arg-Gly-Asp-Ser in the adhesin prompted us to look for a possible binding of E. faecalis cells expressing aggregation substance to eucaryotic cells. We were able to show that the adhesin mediated binding to cultured renal tubular cells (porcine cell line LLC-PK1) via light microscopic, electron microscopic, and enzyme-linked immunosorbent assay-based studies. Synthesis of the adhesin was induced by some component(s) of serum. These data are interpreted to mean that aggregation substance is an adhesin mediating not only cell-cell contact between different E. faecalis strains but also binding of E. faecalis to eucaryotic cells, and therefore it might contribute to virulence.

Amino Acid Sequence↗

PubMed1992

Habitual physical activity in adult IDDM patients. A study with portable motion meters.

The objective of this study was to assess the habitual physical activity of insulin-dependent diabetes mellitus (IDDM) patients on intensive insulin therapy. This case-control study consisted of 34 IDDM patients (14 on multiple injection therapy, 20 on continuous subcutaneous insulin infusion [CSII], and control subjects matched for sex, age, weight, height, body mass index, smoking behavior, and occupational status). Seven IDDM patients were studied before and after changing from multiple injection to CSII therapy. All patients were well controlled according to HbA1c. Portable motion meters were used to assess habitual physical activity during 7 consecutive days with appreciable reproducibility (coefficient of variation 1.24%) and agreement to standardized activity protocols (r = 0.96, P < 0.001). Habitual physical activity was similar in IDDM patients on injection treatment and in controls, respectively. CSII-treated patients exhibited on the average 17% less habitual physical activity than control subjects (P < 0.05). Changing from multiple injection therapy to CSII lowered habitual physical activity insignificantly in seven patients. There was no indication of decreasing physical exercise (e.g., sports) by CSII in this patient group. No correlations were found between habitual physical activity and HbA1c or body mass index, respectively. Habitual physical activity is similar in IDDM patients on multiple injection therapy and control subjects, but may decrease by CSII therapy.

Adult↗

PubMed1991

Genetic transformation of various species of Enterococcus by electroporation.

A transformation system for Enterococcus faecalis was developed which uses untreated (i.e., non-protoplasted) cells and the electroporation technique. The optimized protocol resulted in transformation efficiencies of up to 4 x 10(6) transformants per microgram of plasmid DNA. All strains of E. faecalis tested could be transformed by this method, albeit with differing transformation efficiencies. Using the protocol optimized for E. faecalis we successfully transformed Enterococcus faecium, E. hirae, E. malodoratus and E. mundtii.

Cell Division↗

PubMed1991

Computed tomography as an adjunct to ultrasound in the diagnosis of acute acalculous cholecystitis.

The sonographic and computed tomographic (CT) findings were reviewed in 17 patients with acute acalculous cholecystitis (AAC) over a 6-year period from 1984 to 1989. Of the six patients in whom both ultrasound and CT were performed, CT revealed marked gallbladder (GB) wall abnormalities, including perforation, and pericholecystic fluid collections in five patients not demonstrated by sonography. Of the total group, five patients had GB wall thicknesses of less than or equal to 3 mm (normal) at pathologic examination, which demonstrated a spectrum of disease ranging from acute hemorrhagic/necrotizing, to gangrenous acalculous cholecystitis with perforation. Sonography was falsely negative or significantly underestimated the severity of AAC in seven of the 13 patients examined by sonography. CT because of its superior ability to assess pericholecystic inflammation may provide additional diagnostic information even after a thorough sonographic study in cases of AAC.

Acute Disease↗

PubMed1991

Nucleotide sequence of the gelatinase gene (gelE) from Enterococcus faecalis subsp. liquefaciens.

The gene coding for gelatinase (also called metalloendopeptidase II; microbial proteinase, EC 3.4.24.4) of Enterococcus faecalis subsp. liquefaciens strain OG1-10 was cloned in an Escherichia coli-Enterococcus shuttle vector, and its nucleotide sequence was determined. The DNA sequence encodes one large open reading frame (ORF) with 509 amino acid residues. The ORF contains a signal sequence in its N-terminal region, whereas the N-terminal amino acid sequence determined from the purified extracellular proteinase starts at residue 192 deduced from the ORF. This implies that the gelatinase is synthesized as a prepropolypeptide or prezymogen. The mature gelatinase contains 318 amino acid residues (molecular weight, 34,582) and has significant homology with neutral proteinases from Bacillus species and elastase from Pseudomonas aeruginosa.

Amino Acid Sequence↗

PubMed1991

Comparative analysis of Enterococcus faecalis sex pheromone plasmids identifies a single homologous DNA region which codes for aggregation substance.

An analysis of the 11 known sex pheromone plasmids of Enterococcus faecalis was performed by DNA-DNA hybridization. Plasmids pAD1, pJH2, and pBEM10 turned out to be closely related, whereas pAM373 showed only weak homology with pAD1. A comparison of the hemolysin/bacteriocin determinants of pAD1, pJH2, and pOB1 revealed strong similarities at the DNA level. Our main finding was that one DNA region is conserved among all sex pheromone plasmids, with pAM373 again being an exception; for pAD1 this region was shown earlier to code for aggreagation substance. Detailed hybridization studies of the genes for this plasmid-coded adhesin, which is responsible for cell-cell contact during conjugative transfer via the so-called sex pheromone system of E. faecalis, support the idea of their common origin.

Adhesins, Escherichia coli↗

PubMed1990

Sequence analysis of Enterococcus faecalis aggregation substance encoded by the sex pheromone plasmid pAD1.

The location of the structural gene for aggregation substance on the sex pheromone plasmid pAD1 of Enterococcus faecalis was determined using an oligonucleotide deduced from the N-terminal amino acid sequence of the purified protein. The nucleotide sequence was determined for the corresponding region and two open reading frames (ORFs) could be identified. ORF1 codes for a small (Mr 13,160) acidic protein of unknown function. The gene for aggregation substance (named asa1) was found to code for a protein of 1296 amino acids (Mr 142,248). The protein has a signal peptide of 43 amino acids (the resulting Mr for mature aggregation substance is 137,429) and contains in its C-terminal region a proline-rich sequence, previously characterized as being involved in cell wall association, which is followed by a membrane anchor. The membrane anchor showed significant similarity to that of other Gram-positive organisms, but no other similarities to surface proteins from Gram-positive bacteria were found. In particular, no repeats on the DNA or protein level could be detected for pAD1-specific aggregation substance. The protein contains the amino acid motifs Arg-Gly-Asp-Ser and Arg-Gly-Asp-Val (once each), which, it is proposed, play a crucial role in adherence to eukaryotic cells.

Amino Acid Sequence↗

PubMed1989

Transformation of various species of gram-negative bacteria belonging to 11 different genera by electroporation.

We have undertaken a systematic study to test the transformation of various species of gram-negative bacteria using the electroporation method. The data obtained show very clearly that a great variety of gram-negative bacteria--15 different species belonging to 11 different genera--including freshly isolated wild-type strains can be transformed efficiently by use of the electric-field mediated transformation technique. These include species of the families Enterobacteriaceae, Pseudomonadaceae, Rhizobiaceae, photosynthetic bacteria and strains for which transformation could not be achieved, up to now, by other methods.

Electricity↗

PubMed1989

Identification of aggregation substances of Enterococcus faecalis cells after induction by sex pheromones. An immunological and ultrastructural investigation.

The sex pheromone system of Enterococcus faecalis is responsible for the clumping response of a plasmid carrying donor strain with a corresponding plasmid free recipient strain due to the production of sex pheromones by the recipient strain. The clumping response is mediated by a surface material (called aggregation substance) which is synthesized upon addition of sex pheromones to the cultures. Here we show that after induction a dense layer of "hairlike" structures is formed on the cell wall of the bacteria. These hairlike structures are responsible for the cell-cell contact which leads to the aggregation of cells. Formation of these structures was specific, only occurring after the addition of homologous sex pheromone.

Biological Factors↗

PubMed1989

Localization of aggregation substances of Enterococcus faecalis after induction by sex pheromones. An ultrastructural comparison using immuno labelling, transmission and high resolution scanning electron microscopic techniques.

The distribution of sex pheromone induced aggregation substance was studied on the cell surface of various Enterococcus faecalis strains. In the accompanying paper we have shown that the aggregation substance appears as a layer of hairlike structures. Using direct and indirect immunogold technique, transmission electron microscopy and high resolution scanning electron microscopy we investigated the appearance and distribution of the aggregation substance. The "hairs" increase in number with increasing exposure to sex pheromones (maximum density: 1300/microns2). We show that these structures are unequally distributed over the cell surface, even if the cells were induced by sex pheromones for a long period of time. Statistical analysis of the unequal distribution indicates that aggregation substance is incorporated into pre-existing "old" cell-walls and that this incorporation shows a saturation ca. 40 min after addition of sex pheromones.

Biological Factors↗

PubMed1988

Transformation of bacteria with plasmid DNA by electroporation.

The possibility of electric field-mediated transformation ("electroporation") of a gram-positive bacterium (Enterococcus faecalis) and two gram-negative bacteria (Escherichia coli and Pseudomonas putida) with plasmid DNA was investigated. E. faecalis protoplasts could be transformed by electroporation with a transformation frequency of 10(4) to 10(5) transformants/micrograms plasmid. Untreated--i.e., washed--cells of E. coli could be transformed with rates of 1 X 10(5) transformants/micrograms plasmid DNA. Transformation rates for P. putida cells were up to 3 X 10(4) if the method developed for E. coli was used. Detailed protocols for these systems, including the results of various optimization experiments, are given.

DNA, Bacterial↗

PubMed1988

Transient heart block associated with head trauma.

Atrioventricular nodal block is a rare consequence of head trauma. We report a case of a 63-year-old male with coronary artery disease who sustained a skull fracture. Twelve hours later the cardiac monitor recorded an episode of high-grade atrioventricular block with a ventricular escape rhythm. Subsequent electrophysiologic evaluation revealed no abnormality of atrioventricular nodal conduction nor of intraventricular conduction. Atrioventricular block after head trauma may be caused by enhanced vagal tone.

Creatine Kinase↗

PubMed1986

Studies on the lysosomal binding of 141Ce, 239Np, 239Pu and 241Am in rat and Syrian hamster liver using carrier-free electrophoresis.

The binding of 141Ce, 239Np, 239Pu and 241Am in the liver of rats and Syrian hamsters, following injection in essentially monomeric form, was analyzed by carrier-free electrophoresis at 4-9 days and several months after radionuclide injection. In contrast to density gradient methods lysosomes can be clearly separated from other cell organelles by carrier-free electrophoresis. These, and previous results from this and other laboratories confirm that lysosomes are the main initial binding site for these four radionuclides in the livers of rats and Syrian or Chinese hamster. Light microscopic autoradiography showed that at all the time intervals studied 241Am was more or less uniformly distributed in the liver of all three species. Thus, the changes in the electrophoretic pattern of the nuclides, observed at later time periods in hamsters, cannot be explained by gross redistribution phenomena such as accumulation in macrophages.

Americium↗

PubMed1986

Effective chelation therapy after incorporation of neptunium-239 in rats.

It was demonstrated in rats that it is possible to reduce the retention of 239Np in all body tissues by an early combined treatment with small doses of DTPA and DFOA. The content of 239Np can be decreased in soft tissues even if treatment is delayed. Promptly administered LICAM(C) proved more effective than the above chelate combination in reducing 239Np retention in the bones but increased that in the muscles and especially in the kidneys. This side effect of LICAM(C) could be partly prevented by simultaneous treatment with DTPA.

Animals↗

PubMed1986

Highly efficient protoplast transformation system for Streptococcus faecalis and a new Escherichia coli-S. faecalis shuttle vector.

A highly efficient protoplast transformation system for Streptococcus faecalis has been developed by systematically optimizing different parameters. Up to 10(6) transformants per micrograms of DNA were consistently obtained within 3 days, and cell wall regeneration of protoplasts was virtually 100%. A systematic search for useful vectors showed that the broad-host-range plasmid pIP501 could transform S. faecalis at a high frequency (6.3 X 10(4) transformants per microgram). By combining a high-copy-number derivative of pIP501, designated pGB354, with the Escherichia coli vector pACYC184, we constructed a new E. coli-S. faecalis shuttle vector (pAM401) having nine unique restriction sites. In a shotgun cloning experiment, we ligated a tetracycline resistance determinant from Streptococcus sanguis chromosomal DNA into pAM401 by direct transformation of S. faecalis, establishing the utility of the protoplast transformation system and of the new shuttle vector.

Cloning, Molecular↗

PubMed1985

Identification of transferrin as the principal neptunium-binding protein in the blood serum of rats.

The binding of 239Np(V) to blood serum components of rats was examined in vivo and in vitro. After gel filtration of the serum using a Sephacryl S-300 column, 98% of the applied activity appeared with protein fractions representing coeluted albumins and transferrin. A separation of the albumin- and transferrin-proteins by ion-exchange chromatography using DEAE-cellulose showed the 239Np being entirely bound to the iron-carrier protein transferrin. The high elution yields from the ion-exchange columns, greater than 90%, suggest that the binding may be quite strong. The binding capacity of transferrin for neptunium in vivo was found to decline when the iron level in blood serum was increased. Precipitation experiments showed that 84 +/- 2% of the 239Np was precipitated with 10% (w/v) trichloracetic acid, 77 +/- 3% with 90% ethanol but only 6 +/- 1% with saturated ammonium sulphate at pH 7.4. The available data indicate that as for plutonium, thorium, americium and curium, the iron transport protein, transferrin, may be the main carrier protein for neptunium in mammalian blood serum.

Animals↗

PubMed1985

Gastrointestinal absorption of Np in rats.

The effect of Np mass and the acidity of the administered Np solutions as well as the age, sex and nutritional status of the animals injected or gavaged with 239Np or 237Np were determined. The latter factor proved to be dominant for absorption of Np from the gut. Thus in fasting weanling and young adult male rats, the absorption of 239Np was sixfold higher (0.18% and 0.12%, respectively) than in fed ones (0.03% and 0.02%, respectively). Absorption by fasted adult females was 0.05% of the administered 239Np, about half of that of adult males. Raising the Np-mass gavaged to fasted female rats to 1 and 10 mg 237Np/kg resulted in an absorption of 0.23% and 0.26%, respectively. Thus, an increased absorption of Np in adult rats seems to be expected only if a large mass is ingested. No dependence of the absorption of Np on nitric acid concentration was found. The data obtained after oral administration of 238Pu and 239Np to adult rats suggest that the f1 factor recommended by the ICRP for fractional absorption of soluble Np compounds from the gut should be decreased, whereas the f1 factor for soluble Pu compounds should be raised.

Animals↗