PubMed Health⌕ Search

Biomedical subjects

G Fiedler

Publications and source records attributed to G Fiedler.

At least 19 recordsLinked to original sources

[Renal anemia - an important secondary disease in renal insufficiency].

Anemia is as a frequent complication in patients with chronic kidney disease, which gains in importance in the treatment of patients with renal disease. The main cause of renal anemia is the inadequately low production of endogenous erythropoietin. Often the patients develop an additional absolute or functional iron deficiency, which complicates the diagnostic and therapeutic procedures. Substitution of recombinant human erythropoietin (r-HuEPO) is the most effective therapy. The goal is a stable haemoglobin level >11 g/dl. An often additional existing iron deficiency should be balanced adequately according to the guidelines. With consequent and early treatment morbidity, mortality, and quality of life can be effectively improved.

Anemia↗

[New kinds of relationships on the internet. Virtual object relations in psychotherapy].

Communication in the internet by e-mail is gaining in significance in counseling and psychotherapy of suicidal patients as well as an additional communication in face-to-face psychotherapy. E-Mail contacts during psychodynamic psychotherapy are defined as a transference offer of a virtual object relation. In two case descriptions of suicidal patients, examples of making contact with psychotherapy on the internet are presented. The concept of the virtual object is explained and its application for understanding these initial scenes and phenomena is discussed.

Adult↗

[Pain relief for vaginal delivery--an overview].

The multimodal aspects of labour pain, it's origin and effects, as well as the possibilities of it's treatment will be described from the obstetrician's point of view. This will include a comparison of methods in current common use during delivery including psychosomatic methods, physical methods, acupuncture, homeopathy, use of water, drug therapy as well as the use of local and regional anaesthesia. For particular discussion is the use of epidural catheter in delivery, with respect to availability, optimal timing and obtaining informed consent and it's documentation, as well as teamwork and clear interdisciplinary decisions concerning frequency and parameters of monitoring for epidural catheter anaesthesia following the motto: "how much is necessary, how little is possible?"

Adult↗

NtcA-dependent expression of the devBCA operon, encoding a heterocyst-specific ATP-binding cassette transporter in Anabaena spp.

The devBCA operon, encoding subunits of an ATP-binding cassette exporter, is essential for differentiation of N(2)-fixing heterocysts in Anabaena spp. Nitrogen deficiency-dependent transcription of the operon and the use of its transcriptional start point, located 762 (Anabaena variabilis strain ATCC 29413-FD) or 704 (Anabaena sp. strain PCC 7120) bp upstream of the translation start site, were found to require the global nitrogen transcriptional regulator NtcA. Furthermore, NtcA was shown to bind in vitro to the promoter of devBCA.

ATP-Binding Cassette Transporters↗

Sequence and mutational analysis of the devBCA gene cluster encoding a putative ABC transporter in the cyanobacterium Anabaena variabilis ATCC 29413.

The devBCA gene cluster (dev for development), shown to be essential for envelope formation in heterocysts of Anabaena sp. strain PCC 7120, was identified in the gene bank of a second heterocyst-forming strain, Anabaena variabilis ATCC 29413. Sequence and structural organization of the three genes, encoding subunits of a presumptive ABC transporter, were nearly identical in both strains. Mutants of A. variabilis defective in the devA gene were constructed. As devA mutants of Anabaena 7120, A. variabilis mutants were unable to grow on N2 as sole nitrogen source due to incomplete differentiation of heterocysts.

ATP-Binding Cassette Transporters↗

[Combined myopia-astigmatism correction. A comparison with pure myopia correction].

BACKGROUND: Myopic PRK up to 6 D is an approved and safe procedure. The efficacy and safety of combined treatment of myopia and astigmatism in compound myopic astigmatism (PARK) have not yet been demonstrated. PATIENTS AND METHODS: In a prospective study in 342 eyes the results of myopic PRK were compared to those of correction of compound myopic astigmatism (PARK). RESULTS: Regarding spherical refraction achieved there was no significant difference between groups. Best corrected visual acuity only improved in the group with high astigmatism; this group also showed the least number of Central Islands. Permanent haze and steroid-induced glaucoma or cataract were not observed after treatment up to 6 D. Average astigmatism of 2.24 D was be reduced to 0.53 D 1 year post-operatively. CONCLUSIONS: Correction of astigmatism resulted in undercorrection of approximately 30%. Our experience with the keracor shows PRK up to 6 D with or without additional correction of astigmatism (PARK) to be comparatively safe in terms of regression and complications and equally accurate in reaching spherical target refraction.

Adult↗

The DevBCA exporter is essential for envelope formation in heterocysts of the cyanobacterium Anabaena sp. strain PCC 7120.

The gene devA of the filamentous heterocyst-form-ing cyanobacterium Anabaena sp. strain PCC 7120 encodes a protein with high similarity to ATP-binding cassettes of ABC transporters. Mutant M7 defective in the devA gene is arrested in the development of heterocysts at an early stage and is not able to fix N2 under aerobic conditions. The devA gene is differentially expressed in heterocysts. To gain a better understanding of the structural components of this putative ABC transporter, we determined the complete nucleotide sequence of the entire gene cluster. The two additional genes, named devB and devC, encode proteins with similarities to membrane fusion proteins (DevB) of several ABC exporters and to membrane-spanning proteins (DevC) of ABC transporters in general. Site-directed mutations in each of the three genes resulted in identical phenotypes. Heterocyst-specific glycolipids forming the laminated layer of the envelope were identified in lipid extracts of M7 and in the site-directed mutants. However, transmission electron microscopy revealed unequivocally that the glycolipid layer is missing in mutant M7. Ultrastructural analysis also confirmed a developmental block at an early stage of differentiation. The results of this study suggest that the devBCA operon encodes an exporter of glycolipids or of an enzyme that is necessary for the formation of the laminated layer. The hypothesis is proposed that an intact envelope could be required for further heterocyst differentiation.

ATP-Binding Cassette Transporters↗

The transcriptional activation pattern of lipopolysaccharide binding protein (LBP) involving transcription factors AP-1 and C/EBP beta.

Lipopolysaccharide (LPS) Binding Protein (LBP) is an acute phase protein with the ability to recognize bacterial LPS and transport it to the CD14 molecule or into HDL particles. It is synthesized in hepatocytes and secreted into the blood stream. LBP levels significantly rise during the acute phase response and levels of LBP may be important for an appropriate host reaction to bacterial challenge and for developing the sepsis syndrome. In order to elucidate the mechanisms of LBP regulation we investigated its transcription pattern and performed promoter studies under experimental conditions mimicking an acute phase scenario. In human hepatoma cell lines stimulation with IL-1 beta, IL-6, TNF-alpha and dexamethasone leads to strong transcriptional activation of the LBP gene in a dose- and time-dependent manner. IL-6 alone induces LBP significantly, whereas IL-1 beta mainly increases the IL-6 effect when applied in combination. Our results furthermore show that AP-1 and C/EBP beta are transcription factors involved in the activation of the LBP gene, as revealed by Luciferase reporter gene analysis and electromobility shift assays. Elucidating the mechanism of transcriptional activation of LBP potentially may help in understanding host-pathogen response patterns and mechanisms involved in the acute phase reaction and in the pathophysiology of sepsis.

Acute-Phase Proteins↗

Genetics of a novel starch utilisation pathway present in Klebsiella oxytoca.

A 14.3 kb DNA fragment from Klebsiella oxytoca M5a1 has been cloned and shown to provide Escherichia coli with the capacity for growth on alpha- and beta-cyclodextrins. This fragment is located immediately upstream of the previously identified cgt gene coding for cyclodextrin glycosyltransferase. It contains ten genes (cym) organised in two divergently oriented clusters separated by a non-coding region of 419 bp. Four of the genes code for products homologous to the maltose and linear maltodextrin uptake system, another one for a putative cytoplasmic cyclodextrinase. The cym genes of K. oxytoca are distinct and different from the mal genes; cym mutations do not affect maltose catabolism. On the other hand, whereas mutations in the maltose/maltodextrin-uptake genes do not influence cyclodextrin metabolism, a mutation inactivating the malPQ genes coding for maltodextrin phosphorylase and amylomaltase does. Cyclodextrin catabolism is independent of the presence of a functional cyclodextrin glycosyltransferase but degradation of starch and gamma-cyclodextrins requires the activity of this enzyme. The results indicate the existence of a novel starch degradation pathway which involves the extracellular conversion of starch into cyclodextrins by cyclodextrin glycosyltransferase, uptake of the cyclodextrins by a specific uptake system and intracellular linearisation by a cyclodextrinase. The malto-oligosaccharides produced are then channelled into the maltodextrin-degradation route involving the activity of maltodextrin phosphorylase and amylomaltase.

Amino Acid Sequence↗

Characterization of devA, a gene required for the maturation of proheterocysts in the cyanobacterium Anabaena sp. strain PCC 7120.

Mutant M7, obtained by transposon mutagenesis of the cyanobacterium Anabaena sp. strain PCC 7120, is impaired in the development of mature heterocysts. Under aerobic conditions, the mutant is unable to fix N2 because of a deficiency of at least two components of the oxygen-protective mechanisms: a hemoprotein-coupled oxidative reaction and heterocyst-specific glycolipids. DNA contiguous with the inserted transposon was recovered from the mutant and sequenced. The transposon had inserted itself within a 732-bp open reading frame designated devA. The wild-type form of devA, obtained from a lambda-EMBL3 library of Anabaena sp. DNA, had the identical sequence. Directed mutagenesis of devA in the wild-type strain showed that the phenotype of the mutant was caused by insertion of the transposon. The wild-type form of devA on a shuttle vector complemented the mutation in M7. Expression of devA by whole filaments, monitored following nitrogen stepdown by using luxAB as the reporter, increased ca. eightfold during differentiation; the increase within differentiating cells was much greater. The deduced sequence of the DevA protein shows strong similarity to the ATP-binding subunit of binding protein-dependent transport systems. The product of devA may, therefore, be a component of a periplasmic permease that is required for the transition from a proheterocyst to a mature, nitrogen-fixing heterocyst.

ATP-Binding Cassette Transporters↗

Sequence and transcriptional analysis of the nourseothricin acetyltransferase-encoding gene nat1 from Streptomyces noursei.

We have determined the nucleotide (nt) sequence of nat1, a gene encoding nourseothricin (Nc) acetyltransferase (AT) from Streptomyces noursei, and its transcriptional start point (tsp). The nt sequence upstream from the coding region is completely different from that of the stat gene (encoding streptothricin AT) from Streptomyces lavendulae [S. Horinouchi, K. Furuya, M. Nishiyama, H. Suzuki and T. Beppu, J. Bacteriol. 169 (1987) 1929-1937], even though the nt sequences of the two genes and the deduced amino acid (aa) sequences of the two enzymes show a high degree of similarity. Another stat gene, derived from a Gram-negative plasmid, showed only deduced aa similarity, but not nt sequence similarity, to the above two. A database search for related aa sequences did not reveal any clear-cut homologies to other types of protein. A multiple aa sequence alignment of several ATs is presented.

Acetyltransferases↗

Nucleotide sequence analysis of five putative Streptomyces griseus genes, one of which complements an early function in daunorubicin biosynthesis that is linked to a putative gene cluster involved in TDP-daunosamine formation.

Sequence analysis of the lkmB region of the daunorubicin biosynthetic gene cluster of Streptomyces griseus JA3933 revealed two contiguous open reading frames (ORF) in the same orientation, and three ORFs in the opposite orientation together extending over a 4.6 kb region adjacent to a homologue of the S. peucetius dnrJ gene. ORF1 complemented in trans the lkmB mutation, which seems to affect an early step in daunorubicin biosynthesis. Its deduced product showed no similarity to any known enzyme in the databases. The mutation in ORF1 was localised to a C-T transition at position 1172, leading to the change from a glycine to aspartic acid in the deduced protein. The lack of any homology to known polyketide synthesis enzymes indicates a regulatory role for the product of ORF1, despite the ability of lkmB mutants to further metabolise alkanoic acid. The genes of the oppositely oriented cluster seem to be involved in sugar metabolism. The putative ORF3 protein revealed strong homology to eukaryotic acyl CoA dehydrogenases and might encode an enzyme for the oxidoreduction preceding the introduction of the amino group into daunosamine, and the ORF4 protein is homologous to several epimerases, central enzymes in the formation of the L-2,3,6-trideoxy-3-aminohexoses from TDP-D-glucose. ORF5 seems also to be related to enzymes metabolising nucleotide-activated hexoses.

Amino Acid Sequence↗

Alterations of polymorphonuclear leukocyte glycogen metabolism and glucose uptake in dialysis patients.

Human polymorphonuclear leukocytes (PMNs) are activated during extracorporeal circulation. An indicator of PMN activation may be the glycogen-degrading enzyme phosphorylase. It is unknown whether dialysis therapy may influence PMN carbohydrate metabolism. Therefore, PMNs were isolated from healthy control subjects, patients undergoing continuous ambulatory peritoneal dialysis (CAPD), and patients undergoing regular hemodialysis therapy (RDT) before, during, and at the end of hemodialysis (HD) treatment using dialyzers made of polysulfone or polymethylmethacrylate (PMMA). Nifedipine (NIF) was continuously infused during HD with PMMA in 5 patients at a dose of 18 micrograms/kg body weight per hour. Glycogen, activity of glycogen synthetase and phosphorylase (active and inactive forms of both enzymes), and glucose uptake with and without stimulation with the chemotactic peptide FMLP were determined in these PMNs. During HD with PMMA, there was a significant increase of PMN phosphorylase "a" activity 15 and 30 minutes after the start of HD. HD with polysulfone did not stimulate the active "a" form of the glycogen-degrading enzyme in PMNs. HD with PMMA significantly inhibited the active I-form of glycogen synthetase, whereas polysulfone activated glycogen synthetase I. NIF inhibited phosphorylase "a" activation during HD with PMMA. PMN glycogen content and glucose uptake were improved during HD with polysulfone, but not with PMMA. PMN glycogen content, activity of glycogen synthetase, and glucose uptake were significantly lower also in CAPD patients compared with healthy controls. These data show that HD with PMMA activates PMN glycogenolysis. This effect can be inhibited by calcium channel blockers. PMN glycogen content of RDT and CAPD patients is significantly lower compared with healthy controls due to inhibition of glycogen synthesis. Elimination of dialyzable factor(s) improves, but does not restore, PMN glycogen synthesis and glucose uptake.

Adult↗

Arsenical resistance of growth and phosphate control of antibiotic biosynthesis in Streptomyces.

Twenty-six wild-type Streptomyces strains tested for resistance to arsenate, arsenite and antimony(III) could be divided into four groups: those resistant only to arsenite (3) or to arsenate (2) and those resistant (8) or sensitive (13) to both heavy metals. All strains were sensitive to antimony. The structural genes for the ars operon of Escherichia coli were subcloned into various Streptomyces plasmid vectors. The expression of the whole ars operon in streptomycetes may be strain-specific and occurred only from low-copy-number plasmids. The arsC gene product could be expressed from high-copy plasmids and conferred arsenate resistance to both E. coli and Streptomyces species. The ars operon expressed in S. lividans and the arsC gene expressed in S. noursei did not render the synthesis of undecylprodigiosin and nourseothricin, respectively, phosphate-resistant. In addition in wild-type strains of Streptomyces phosphate sensitivity of antibiotic biosynthesis did not show strong correlation with resistance of growth to arsenicals.

Anti-Bacterial Agents↗

Analysis of the nourseothricin-resistance gene (nat) of Streptomyces noursei.

A gene (nat) conferring resistance to the streptothricin antibiotic nourseothricin (Nc) was cloned from the producer Streptomyces noursei into Streptomyces lividans on the vector pIJ702 to form pNAT1. The nat gene was localized on a 1-kb SalI-MboI fragment, which also carries the nat promoter. Divergent promoter activity from the nat promoter region was identified on the cloned fragment using promoter probe plasmids pIJ486 and pIJ487. The nat gene is not expressed from its own promoter in Escherichia coli as shown by its failure to promote cat expression in promoter-less plasmid pBB100 and by the expression of NcR in only one orientation, when cloned in pUC19. In S. lividans 7A, harbouring plasmid pNAT1, an Nc-acetylating activity (NAT) was associated with the cloned resistance gene. The substrate specificity of NAT correlated well with the substrate range of the acetyltransferase in S. noursei and Tn1825-determined streptothricin resistance in Gram-negative bacteria. Moreover, an extract of S. lividans carrying pNAT1 showed specific serological cross-reactivity with an extract of E. coli carrying Tn1825.

Acetyltransferases↗

Copy number mutants of the broad-host-range Streptomyces plasmid pMG200.

pMG200, isolated from the bacteriocin-releasing strain Streptomyces chrysomallus, was further physically mapped. Variants of S. chrysomallus were isolated which inhibited the parental strain. Two types of plasmids, pMG210 and pMG220, were isolated from these variants, with copy numbers of 10-30 and 300, respectively, compared with 1-3 for pMG200. pMG210 is apparently physically identical to pMG200 but presumably differs at a level not detected by simple restriction mapping; pMG220 is deleted for 1.6 kb. Genes for thiostrepton and viomycin resistance were subcloned from pIJ364 on to pMG200 and a fragment containing the gene for nourseothricin resistance was subcloned on to pMG220. In this way nonessential sites were identified.

Cloning, Molecular↗

Gene expression after transformation of Streptomyces lividans protoplasts with plasmid pIJ2 DNA.

A new method has been developed for the selection of antibiotic-resistant clones after transformation of Streptomyces protoplasts with plasmid DNA. This method is based on establishing a spatial concentration gradient for the antibiotic, the resistance to which is encoded by the transforming plasmid. By this method, the resistance development of regenerating protoplasts can be followed. The results suggest that antibiotic resistance is inducible. In addition, we were able to show that resident plasmids incompatible with the incoming ones are eliminated when this direct selection principle is used. Moreover, this method, which may facilitate the application of gene technology in Streptomyces, works even though the transformation procedure gives variable results.

DNA, Bacterial↗