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Biomedical subjects

R Böhm

Publications and source records attributed to R Böhm.

At least 19 recordsLinked to original sources

[Toxic myopathy with kidney failure as a colchicine side effect ifn familial Mediterranean fever].

A 24-year-old woman with familial Mediterranean fever (FMF) had for one year been treated with colchicine, 1 mg daily, for repeated bouts of fever, abdominal pain and arthritis. She was also known to have renal amyloidosis. Lately she had developed gastrointestinal symptoms, muscle pain and obvious, predominantly proximal muscular weakness in both legs. The cause of the symptoms was rhabdomyolysis with an increased creatinine activity of 1000 U/l and marked myoglobinuria (1600 micrograms/l), as well as renal failure with normal uric acid and a creatinine clearance of 3 ml/min per 1.73 m2. Serum creatinine concentration was 970 mumol/l, urea 34 mmol/l. Muscle biopsy corresponded to a subacute necrotizing myopathy with vacuole formation, signs typical of toxic damage. Renal biopsy confirmed advanced amyloidosis. The colchicine dose was reduced to 0.5 mg/d. The renal failure responded to conservative treatment. The myopathy symptoms receded within 4 weeks, creatinine clearance rising to 25 ml/min per 1.73 m2. 12 months after reduction of the colchicine dose the patient was without any FMF-related symptoms.

Acute Kidney Injury

[Newer information about several aspects of animal epidemic disinfection].

The amendment of the German Veterinary Legislation concerning the eradication of notifiable diseases demanded also an updating of the relevant disinfection measures and technologies. Some hints are given, based on own experimental results, for the choice of chemical disinfectants, on some special problems of disinfection in livestock buildings and of disinfection of animal slurry and farm-yard manure as well as on biotechnological disinfection methods and environment-friendly disinfectants.

Animals

Mutational analysis of the operon (hyc) determining hydrogenase 3 formation in Escherichia coli.

In-frame deletions were introduced into each of the eight genes of the hyc operon coding for products required for the formation of the formate hydrogenlyase (FHL) system. The deletions were transferred to the chromosome and the resulting mutants were analysed for development of formate dehydrogenase H and hydrogenase 1, 2 and 3 activity. It was found that hycA, the promoter-proximal gene, is a regulatory gene and that it codes for a product counteracting transcriptional activation by FhlA. Deletions within the hycB to hycH genes specifically affected formate dehydrogenase H activity or hydrogenase 3 activity, or both. None of the mutations affected hydrogenase 1 or 2 activity. A model is proposed for the functional interaction of the different hyc operon gene products in the formate hydrogenlyase complex, which is based on the results of the mutational analysis, on the determination of the subcellular localization of the FdhF, HycE, HycF and HycG polypeptides and on the similarity of hyc gene product sequences with those from other hydrogenase systems. HycH, the product of the most promoter-distal gene, does not seem to form part of the functional FHL complex but rather is required for the conversion of a precursor form of the large subunit of hydrogenase 3 into the mature form.

Amino Acid Sequence

[Research in veterinary environmental hygiene].

The course of development of the initial subject "Veterinary Hygiene" to the reduction and specialization limited to "Animal Hygiene" and the recent extension into the broad field of environmental hygiene is described. This is done with reference to books and other publications which point the way ahead for research of veterinary medicine in environmental hygiene and thus prove the increasing importance of that subject. A survey is given over current German research in that field, future demands as well as the necessary extension of environmental hygiene in graduate and post-graduate teaching of veterinary students as well as further professional training of veterinarians.

Animals

Molecular characterization of an operon (hyp) necessary for the activity of the three hydrogenase isoenzymes in Escherichia coli.

The 58/59 min region of the Escherichia coli chromosome contains two divergently oriented gene clusters coding for proteins with a function in hydrogenase formation. One cluster (the hyc operon), transcribed counterclockwise with respect to the E. coli chromosome, codes for gene products with a structural role in hydrogenase 3 formation (Böhm et al., 1990). The nucleotide sequence of the divergently transcribed operon (hyp) has been determined. It contains five genes, all of which are expressed in vivo in a T7 promoter/polymerase system, and the sizes of the synthesized products correspond with those predicted from the amino acid sequence. Complementation analysis of previously characterized mutants showed that the hypB, hypC and hypD genes have a function in the formation of all three hydrogenase isoenzymes, lesions in hypB being complemented by high nickel ion concentration in the medium. Prevention of hypBCDE gene expression led to an altered electrophoretic pattern of hydrogenase 1 and 2 constituent subunits, indicating increased chemical or proteolytic subunits, Under fermentative growth conditions, operon expression was governed by an NtrA-dependent promoter lying upstream of hypA working together with an fnr gene product-dependent promoter which was localized within the hypA gene. The latter (operon-internal) promoter is responsible for hypBCDE transcription under non-fermentative conditions when the -24/-12 NtrA-dependent promoter upstream of hypA is silent.

Amino Acid Sequence

[The possibility of the serological differentiation of Clostridium species using enzyme immunoassays].

A method for serotyping and diagnosis of 30 selected Clostridia strains of 12 different species by an enzyme-immuno-assay using polystyrene balls as a solid phase is described and tested for its efficiency. The reference strains and the homologous test strains could be differentiated from the heterologous in principle. A reliable detection of the several serotypes of Cl. perfringens was not perfectly to achieve with the conjugates used here, cross-reaction with Cl. chauvoei could be removed by absorption. The conjugate of Cl. bifermentans reacted with Cl. sordellii and Cl. difficile too, and must be absorbed prior any diagnostic applications with vegetative cells of the corresponding strains. Concerning the antigenic relationships between the Clostridia strains used here, the results of Ellner and Green (7, 8) as well as of Poxton and Byrne (14) could be reconfirmed. Moreover the applied technique offers the possibility to come to a more detailed result concerning the degree of antigenic relationship. The developed method is a valuable tool for the differentiation of pathogenic Clostridia and within the group Cl. bifermentans/Cl. sordellii/Cl. difficile a rapid diagnosis at the species level could be achieved. Some limitations by cross-reactions are given, but results can be improved by absorption of the conjugates.

Clostridium

The effect of verapamil on carotid artery distensibility and cross-sectional compliance in hypertensive patients.

The effect of chronic administration of verapamil, following a double-blind, crossover protocol, on the distensibility and cross-sectional compliance of the common carotid artery was investigated in 19 patients with essential hypertension. Distensibility was significantly increased during verapamil treatment as compared to placebo for both left and right common carotid artery (p less than 0.01 and p less than 0.05, respectively). The cross-sectional compliance was significantly increased during verapamil treatment for the right common carotid artery (p less than 0.05). For the left common carotid artery, the difference did not reach the level of significance (p = 0.16). The common carotid artery diameter and the arterial pulse pressure were not significantly different during verapamil treatment as compared with placebo. The results of this study indicate that chronic treatment with verapamil increases distensibility and cross-sectional compliance of the common carotid artery in hypertensive subjects. These improvements have to be considered as changes in arterial wall properties because no significant differences in pulse pressure and diameter could be detected between the verapamil and placebo periods. Improved arterial wall properties could result in a better management of the flow jet from the heart, and might protect the patients from atherosclerotic complications of hypertension.

Adult

Nucleotide sequence and expression of an operon in Escherichia coli coding for formate hydrogenlyase components.

An 8kb segment of DNA from the 58/59 min region of the E. coli chromosome, which complements the defect of a mutant devoid of hydrogenase 3 activity, has been sequenced. Eight open reading frames were identified which are arranged in a transcriptional unit; all open reading frames were transcribed and translated in vivo in a T7 promoter/polymerase system. Analysis of the amino acid sequences derived from the nucleic acid sequences revealed that one of them, open reading frame 5 (ORF5), exhibits significant sequence similarity to conserved regions of the large subunit from Ni/Fe hydrogenases. Two of the open reading frames (orf2, orf6) code for proteins apparently carrying iron-sulphur clusters of the 4Fe/4S ferredoxin type. The product of one of the open reading frames, orf7, displays extensive sequence similarity with protein G from the chloroplast electron transport chain. ORF3 and ORF4, on the other hand, are extremely hydrophobic proteins with nine and six putative transmembrane helices, respectively. Over a limited hydrophilic sequence stretch, bordered by putative transmembrane areas, ORF3 and ORF4 exhibit homology with subunits 4 and 1 of mitochondrial and plastid NADH-ubiquinol oxidoreductases, respectively. The operon described, therefore, appears to comprise genes for redox carriers linking formate oxidation to proton reduction and for a hydrogenase of hitherto unique composition.

Aldehyde Oxidoreductases

Characterization of divergent NtrA-dependent promoters in the anaerobically expressed gene cluster coding for hydrogenase 3 components of Escherichia coli.

The regulatory region of two divergently oriented transcriptional units involved in the formation of the gas-evolving hydrogenase (isoenzyme 3) of Escherichia coli was investigated. DNA sequence analysis revealed the existence of a 210 bp non-coding region containing two sequences showing homology to -24/-12 NtrA-dependent promoters. These sequences were arranged in a divergent orientation entirely consistent with their being involved in transcribing the divergent operons. Through S1 protection experiments it could be shown that transcription of both promoters was NtrA-dependent and that it was regulated in an identical manner: oxygen repressed expression, as did anaerobic growth in the presence of nitrate; transcription was induced in cells grown anaerobically in the absence of exogenous electron acceptors and formate was found to be obligately required for this anaerobic induction. Lying at an approximately equal distance between both promoters was a short stretch of DNA which showed similarity to the sequence previously identified (Birkmann and Böck, 1989a) as being necessary for formate induction of the fdhF gene.

Anaerobiosis

[Detection specificity of an optimal solid-phase enzyme immunoassay for Pseudomonas aeruginosa and Pseudomonas mallei].

The evaluation and application of an enzyme-immunoassay (EIA) for the detection of Pseudomonas (Ps.) aeruginosa and Ps. mallei is described. Polystyrene beads (1/4'') as the solid-phase are prepared by coating the balls with purified IgG from the serum of rabbits (9-12 micrograms/bead) in Coating-Buffer pH 9.6. After washing the balls they are saturated with 10% BSA or 10% FCS in PBS-Tween 20. The bacteria bound to the coated balls are detected by the specific peroxidase labelled IgG. This EIA using Ps. aeruginosa (P9) as a model is able to detect this bacterium within 5 hours, with stored coated balls 3.5 hours, with a detection limit of 10(4) CFU. Nine Pseudomonas-strains react stronger than other strains. These cross-reactions can be substantially reduced by absorbing the P9-conjugate with the cells of Ps. stutzeri (P15). With the other Pseudomonas-strains a high specificity is found with the P9-conjugate. After modifying this EIA for the detection of Ps. mallei (P18) the strains Ps. mallei (P57), Ps. pseudomallei (P17) and Ps. cepacia (P67) react with the P18-conjugate. With the other tested strains a high specificity is found at 10(7) CFU. The polystyrene bead-EIA is recommended as a sensitive and specific test for the detection of Ps. aeruginosa in about 5 resp. 3.5 hours. It only requires normal laboratory equipment and is thus a highly practicable method for routine diagnostic of Ps. aeruginosa.

Cross Reactions

Pharmacokinetics of nitrendipine in terminal renal failure.

The pharmacokinetics and plasma protein binding of nitrendipine in patients with terminal renal failure have been compared with those in subjects with normal renal function. Kinetic parameters were calculated after a single 40 mg oral dose, an i.v. injection of 3 mg and after a 15 mg i.v. infusion of nitrendipine. Steady-state plasma levels were determined after 5 days of oral treatment with 20 mg b.d. Pharmacokinetic parameters and steady-state plasma levels in patients with renal failure did not differ from those in subjects with normal renal function. Nitrendipine was as highly bound to plasma proteins in patients with renal failure, as in subjects with normal renal function. The plasma protein did not differ between the two. The dosage of nitrendipine need not be modified for kinetic reasons in patients with renal failure.

Administration, Oral

Total and free steady-state plasma levels and pharmacokinetics of nifedipine in patients with terminal renal failure.

The total and free steady-state plasma levels of nifedipine in patients with renal failure have been compared with those in subjects with normal renal function. Studies were done after administration of nifedipine 10 mg t.d.s. p.o. for 5 days, after i.v. infusion of 4.4 mg, and after a single 10 mg oral dose. The systemic clearance of nifedipine after a single i.v.-dose was higher in subjects with renal insufficiency (854 ml/min) than in those with normal renal function (468 ml/min). After the single oral dose the AUC (6100 ng.min.ml-1) and maximum plasma concentration (75.0 ng.ml-1) were lower than in subjects with normal renal function (19300 ng.ml-1; 122 ng.ml-1). The plasma protein binding of nifedipine averaged 95.5% in normal subjects and 94.8% in patients with renal failure. Although free and total steady-state plasma levels of nifedipine tended to be somewhat lower than normal in renal failure, the changes in pharmacokinetics and decreased protein binding of nifedipine did not result in a significantly different steady-state plasma level of the drug. The blood pressure response to a given plasma nifedipine level appeared to be enhanced in renal failure.

Administration, Oral

The safety of blood component preparation with blood cell separators.

Despite initial skepticism following the introduction of cytapheresis with blood cell separators, this technology is now accepted in the field of blood donation. Serious, life-threatening reactions have not been observed and thus could not influence the development, and minor problems can be reduced further on the basis of recommended safety guidelines. Additionally, the continuous registration of side effects is helpful for a complete elimination of undesired reactions.

Blood Cells

[Comparison of the sporocidal effects of two different disinfectants i a model experiment with pathogenic and nonpathogenic clostridial spore as well as with Bacillus cereus].

Official methods concerning the examination of sporocidal effects of disinfectants are presently lacking in the Federal Republic of Germany. Spore resistance of selected clostridial species towards formaldehyde and peracetic acid was examined by suspension tests and germ-carrier tests in comparison to spores of Bacillus cereus. Formaldehyde in concentrations between 5% and 20% as well as peracetic acid in concentrations between .004% and .125% were tested. Spore concentration of 10(6) spores/ml in the suspension-tests and 10(4) spores/ml in the germ-carrier tests with wooden germ-carriers were used. Against formaldehyde spores of Cl. septicum had the lowest resistance in the suspension tests followed by Cl. difficile, Cl. tertium, Cl. bifermentans and Cl. perfringens. Cl. sporogenes strain NCTC 532 had the most resistant spores. Concerning peracetic acid the situation was nearly the same. The resistance of all spores of pathogenic species under investigation towards chemical disinfectants was covered by those of Cl. sporogenes NCTC 532 treated with isopropanol. Isopropanol treated spores of B. cereus offer an alternative solution for practical disinfectant testing because spore production as well as growth of clostridia entail great expense. In order to cover the resistance of spores of Cl. perfringens A, the most resistant pathogenic clostridial species tested here, increased spore concentrations of B. cereus would be necessary.

Bacillus cereus

[The detection of Bacillus anthracis protective antigens by enzyme immunoassay (EIA) using polyclonal and monoclonal antibodies].

An Enzyme-Immunoassay (EIA) for the detection of Bacillus anthracis-protective antigen (PA) within one hour was developed. If the rabbit antiserum was used, 15 ng PA/ml could be detected and with the monoclonal antibody, the detection limit was 60 ng PA/ml. With respect to the higher specificity and with regard to the aspects of animalcare, monoclonal antibodies should be used in the test instead of the polyclonal antiserum.

Animals

[Advice as to the safety precautions for handling infectious material in a veterinary practice laboratory].

The number of microbiological investigations with infectious material increases in veterinary practice. Frequently the working space is cramped so that special security measures are imperative. These measures are discussed in detail. Their disregard can result in laboratory infections with zoonotic agents and in unpleasant lawsuits with bad consequences. The German regulations pertaining to security in laboratories are listed. They should be observed in any case.

Animals

The influence of antituberculosis drugs on the plasma level of verapamil.

The influence of antituberculosis drugs on the plasma level of verapamil was studied after its oral and intravenous administration. Six patients who had been treated for at least 6 months with a combination of rifampicin, ethambutol and isoniazid received a single oral dose of 40 mg verapamil. As compared to untreated subjects, the antituberculosis drugs greatly reduced the bioavailability of the calcium antagonist. Studies in patients in whom treatment with ethambutol and isoniazid had been discontinued revealed that the effect was due to rifampicin. The drugs for tuberculosis had no influence on the plasma level of verapamil when it was given intravenously. The findings can be explained by the induction of verapamil metabolizing liver enzymes in patients treated with rifampicin.

Administration, Oral