Biomedical subjects

G Klug

Publications and source records attributed to G Klug.

At least 19 recordsLinked to original sources

PubMed2006

Evaluation of transient apical ballooning with cardiac magnetic resonance imaging and 31-phosphorous magnetic resonance spectroscopy.

Apical ballooning is an increasingly reported transient cardiomyopathy with yet unknown origin. In this study 2 cases of apical ballooning are described in whom we used a combined approach of cardiac magnetic resonance imaging (CMR) and 31-Phosphorous magnetic resonance spectroscopy (31P MRS). Electrocardiogram showed ST abnormalities and cardiac serum markers were mildly elevated, but CAG demonstrated smooth coronary arteries. Cine-CMR revealed severe apical akinesia and significantly decreased ejection fraction. Furthermore we detected reduced myocardial phosphocreatine to beta-ATP (PCr/b-ATP) ratios during the first week of acute disease. After 1 week we observed an improvement of PCr/b-ATP ratios by 68% and 34%, which was associated with an increase in left ventricular function. Our data suggest that 31P MRS might be a valuable tool in the evaluation of apical ballooning, but larger cohorts are needed to improve the understanding of metabolic changes during transient apical ballooning.

Aged↗

PubMed2005

[Change in social networks due to psychoses].

The paper presents the network concept and filters research results according to practical relevance. By way of introduction, the concept of the social network SNW is briefly outlined in terms of its origins and significance. The "socially intact" network of 25 - 40 individuals, of which 15 - 20 % are very close and familiar, is compared with that of people suffering from schizophrenia, averaging 13 individuals. The changes are compiled according to the individuals involved, the type of relationship, and the type of perception. Outcome-relevant results of the network research are summarised on a general basis and according to the parameters social support, social and family contacts, distribution by sex, symptoms, and relatives. The changes in the network in the course of the disease and their importance are described separately according to the phases premorbid phase, prodromal phase, early psychosis, untreated psychosis, first phase of treatment, and long-term phase, illustrating their importance in the context of the overall pathological process. Possibilities of intervention in the SNW are outlined for the phases premorbid, prodromal, onset of the first episode, chronicity development, and existing chronicity. The conclusion rounds off with a brief enumeration of the often lacking intervention options in the German-speaking world.

Family Relations↗

PubMed2002

Bacteriochlorophyll-dependent expression of genes for pigment-binding proteins in Rhodobacter capsulatus involves the RegB/RegA two-component system.

Expression of the puf and puc operons, which encode proteins of the photosynthetic apparatus of Rhodobacter capsulatus, is regulated by oxygen. A drop in the oxygen tension in the environment leads to an increase in the levels of puf and puc mRNAs. In strains lacking bacteriochlorophyll (Bchl) due to mutations in bch genes, the rise in puf and puc mRNA levels observed on reduction of oxygen tension is much less pronounced than in wild-type cells, indicating co-regulation of the syntheses of pigments and pigment-binding proteins. Here we show that Bchl synthesis also affects the expression of the bchC gene, which codes for a subunit of bacteriochlorophyll synthase, suggesting an autoregulatory mechanism for the Bchl biosynthetic pathway. Furthermore, our data provide evidence that the RegB/RegA two-component system, which is known to play a central role in oxygen-controlled expression of photosynthesis genes, is also involved in the Bchl-dependent regulation. Mutant strains which do not synthesize RegB or RegA show similar oxygen-dependent puf and puc expression in the presence and absence of Bchl. Our results support the view that the RegB/RegA system can directly or indirectly sense whether Bchl synthesis takes place or not.

Bacterial Proteins↗

PubMed2001

Both N-terminal catalytic and C-terminal RNA binding domain contribute to substrate specificity and cleavage site selection of RNase III.

The double-stranded RNA-specific endoribonuclease III (RNase III) of bacteria consists of an N-terminal nuclease domain and a double-stranded RNA binding domain (dsRBD) at the C-terminus. Analysis of two hybrid proteins consisting of the N-terminal half of Escherichia coli RNase III fused to the dsRBD of the Rhodobacter capsulatus enzyme and vice versa reveals that both domains in combination with the particular substrate determine substrate specificity and cleavage site selection. Extension of the spacer between the two domains of the E. coli enzyme from nine to 20 amino acids did not affect cleavage site selection.

Base Sequence↗

PubMed2001

An mRNA degrading complex in Rhodobacter capsulatus.

An RNA degrading, high molecular weight complex was purified from Rhodobacter capsulatus. N-terminal sequencing, glycerol-gradient centrifugation, and immunoaffinity purification as well as functional assays were used to determine the physical and biochemical characteristics of the complex. The complex comprises RNase E and two DEAD-box RNA helicases of 74 and 65 kDa, respectively. Most surprisingly, the transcription termination factor Rho is a major, firmly associated component of the degradosome.

Amino Acid Sequence↗

PubMed2001

A randomized trial of very early decompressive craniectomy in children with traumatic brain injury and sustained intracranial hypertension.

OBJECT: The object of our study was to determine, in children with traumatic brain injury and sustained intracranial hypertension, whether very early decompressive craniectomy improves control of intracranial hypertension and longterm function and quality of life. METHODS: All children were managed from admission onward according to a standardized protocol for head injury management. Children with raised intracranial pressure (ICP) were randomized to standardized management alone or standardized management plus cerebral decompression. A decompressive bitemporal craniectomy was performed at a median of 19.2 h (range 7.3-29.3 h) from the time of injury. ICP was recorded hourly via an intraventricular catheter. Compared with the ICP before randomization, the mean ICP was 3.69 mmHg lower in the 48 h after randomization in the control group, and 8.98 mmHg lower in the 48 hours after craniectomy in the decompression group (P=0.057). Outcome was assessed 6 months after injury using a modification of the Glasgow Outcome Score (GOS) and the Health State Utility Index (Mark 1). Two (14%) of the 14 children in the control group were normal or had a mild disability after 6 months, compared with 7 (54%) of the 13 children in the decompression group. Our conclusion was that when children with traumatic brain injury and sustained intracranial hypertension are treated with a combination of very early decompressive craniectomy and conventional medical management, it is more likely that ICP will be reduced, fewer episodes of intracranial hypertension will occur, and functional outcome and quality of life may be better than in children treated with medical management alone (P=0.046; owing to multiple significance testing P <0.0221 is required for statistical significance). This pilot study suggests that very early decompressive craniectomy may be indicated in the treatment of traumatic brain injury.

Brain Injuries↗

PubMed2000

The development of agoraphobia in panic disorder: a predictable process?

BACKGROUND: Panic attacks are conceptualized to be the central feature of both panic disorder without (PDU) and with agoraphobia (PDA). As a sizeable percentage of panic patients do not develop avoidance behavior, other factors than 'panic attacks', in general, must influence the different courses of the disorder. METHOD: We studied 84 outpatients suffering from PDU or PDA concerning different factors which were hypothesized to influence the development of agoraphobia. RESULTS: 'Earlier age of onset', 'fear of losing control' and 'chills or hot flushes' turned out to correlate statistically significantly with PDA, while 'chest pain or discomfort' occurred more often in PDU. LIMITATIONS: The present study used retrospective data. CONCLUSIONS: The results of this study suggest that the development of agoraphobia in panic disorder is influenced by specific variables and is not a purely coincidental process.

Adult↗

PubMed2000

Initial events in the degradation of the polycistronic puf mRNA in Rhodobacter capsulatus and consequences for further processing steps.

Individual segments of the polycistronic puf mRNA of Rhodobacter capsulatus exhibit extremely different half-lives contributing to the stoichiometry of light-harvesting and reaction centre complexes of this facultative phototrophic bacterium. While earlier investigations shed light on the processes leading to the degradation of the 2.7 kb pufBALMX mRNA and, consequently, to the formation of the highly stable 0.5 kb pufBA mRNA processing product, we have now investigated the initial events in the degradation of the highly unstable 3.2 kb pufQBALMX primary transcript. Sequence modifications of two putative RNase E recognition sites within the pufQ coding region provide strong evidence that RNase E-mediated cleavage of a sequence at the 3' end of pufQ is involved in rate-limiting cleavage of the primary pufQBALMX transcript in vivo. The putative RNase E recognition sequence at the 5' end of pufQ is cleaved in vitro but does not contribute to rate-limiting cleavage in vivo. Analysis of the decay of puf mRNA segments transcribed from wild-type and mutated puf DNA sequences in R. capsulatus and Escherichia coli reveal that RNase E-mediated cleavage within the pufQ mRNA sequence also affects the stability of the 0.5 kb pufBA processing product. These findings demonstrate that the stability of a certain mRNA segment depends on the pathway of processing of its precursor molecule.

Bacterial Proteins↗

PubMed2000

RNase III processing of intervening sequences found in helix 9 of 23S rRNA in the alpha subclass of Proteobacteria.

We provide experimental evidence for RNase III-dependent processing in helix 9 of the 23S rRNA as a general feature of many species in the alpha subclass of Proteobacteria (alpha-Proteobacteria). We investigated 12 Rhodobacter, Rhizobium, Sinorhizobium, Rhodopseudomonas, and Bartonella strains. The processed region is characterized by the presence of intervening sequences (IVSs). The 23S rDNA sequences between positions 109 and 205 (Escherichia coli numbering) were determined, and potential secondary structures are proposed. Comparison of the IVSs indicates very different evolutionary rates in some phylogenetic branches, lateral genetic transfer, and evolution by insertion and/or deletion. We show that the IVS processing in Rhodobacter capsulatus in vivo is RNase III-dependent and that RNase III cleaves additional sites in vitro. While all IVS-containing transcripts tested are processed in vitro by RNase III from R. capsulatus, E. coli RNase III recognizes only some of them as substrates and in these substrates frequently cleaves at different scissile bonds. These results demonstrate the different substrate specificities of the two enzymes. Although RNase III plays an important role in the rRNA, mRNA, and bacteriophage RNA maturation, its substrate specificity is still not well understood. Comparison of the IVSs of helix 9 does not hint at sequence motives involved in recognition but reveals that the "antideterminant" model, which represents the most recent attempt to explain the E. coli RNase III specificity in vitro, cannot be applied to substrates derived from alpha-Proteobacteria.

Alphaproteobacteria↗

PubMed2000

Correction of the DNA sequence of the regB gene of Rhodobacter capsulatus with implications for the membrane topology of the sensor kinase regB.

We corrected the previously published sequence for the regB gene, which encodes a histidine sensor kinase in Rhodobacter capsulatus. The deduced RegB amino acid sequence has an additional putative transmembrane domain at the N terminus. Analysis of RegB-PhoA and RegB-LacZ fusion proteins supports a topology model for RegB with six membrane-spanning domains.

Alkaline Phosphatase↗

PubMed2000

In vivo and in vitro analysis of RegA response regulator mutants of Rhodobacter capsulatus.

In the facultative photosynthetic bacterium Rhodobacter capsulatus, the transcription of genes encoding pigment binding proteins is tightly regulated in response to the oxygen partial pressure by the RegB/ RegA two component system. After a shift from high to low oxygen tension, the response regulator RegA enhances transcription of the puf and puc operon coding for the reaction center, light-harvesting complex I (LHI), and LHII proteins. Various regA mutant strains were analyzed in this study. In a RegA deficient strain, activation of puf and puc transcription is severely impaired which consequently leads to the synthesis of only a few photosynthetic complexes. Strains carrying a mutation in the helix-turn-helix domain of RegA or a mutation of the phosphorylation site, Asp63, show a phenotype like the RegA deficient mutant, although the RegA(D63K) mutant protein showed the same DNA binding behavior as the wild type protein. In contrast, the puf and puc mRNAs still reach about 50-70 % of the wild type level after reduction of oxygen tension in strains which synthesize the C-terminal RegA activator domain only or a hybrid protein composed of the RegA activator and the FixJ receiver domain, while both mutant proteins are impaired in DNA binding. Our data suggest that phosphorylation is not required for DNA binding but rather plays a role for efficient initiation of transcription.

Amino Acid Sequence↗

PubMed2000

DNA binding of wild type RegA protein and its differential effect on the expression of pigment binding proteins in Rhodobacter capsulatus.

The transcription of genes encoding pigment binding proteins in the facultative photosynthetic bacterium Rhodobacter capsulatus is regulated in response to oxygen partial pressure. Previous results identified RegA and RegB as members of a two component system involved in oxygen dependent synthesis of the photosynthetic apparatus. Here we demonstrate that RegA differentially controls the transcription of the puf and pucoperons which encode proteins of the LHI and LHII antenna complexes, respectively. In a regA mutant strain the level of puf specific mRNA reaches about 30% of the wild type levels and transcription is still responsive to oxygen tension. In contrast, the level of puc specific mRNA is very low and is no longer oxygen regulated. RegA binds to DNA sequences upstream of both the puf and puc operons, although with different affinities. We provide experimental evidence that a putative helix-turn-helix motif in the C-terminal region of RegA is responsible for its specific binding to the puf and puc promoter regions. In contrast to many other response regulators, the affinity of RegA for the target DNA is only slightly modified by phosphorylation.

Bacterial Proteins↗

PubMed1999

Regulation of bacterial photosynthesis genes by oxygen and light.

Most bacteria have the capability to adapt to changes in their environment. Facultatively phototrophic bacteria like Rhodobacter can switch from aerobic respiration to anoxygenic photosynthesis in the absence of oxygen. The formation of the photosynthetic apparatus is primarily regulated by oxygen tension. The amount of photosynthetic complexes is influenced by the light intensity in anaerobic cultures. This review focuses on the molecular mechanisms involved in the regulation of Rhodobacter photosynthesis genes by oxygen and light.

Gene Expression Regulation, Bacterial↗

PubMed1999

The amyloid beta-protein precursor of Alzheimer's disease is degraded extracellularly by a Kunitz protease inhibitor domain-sensitive trypsin-like serine protease in cultures of chick sympathetic neurons.

The amyloid beta-protein precursor (APP) of Alzheimer's disease (AD) is cleaved either by alpha-secretase to generate an N-terminally secreted fragment, or by beta- and gamma-secretases to generate the beta-amyloid protein (Abeta). The accumulation of Abeta in the brain is an important step in the pathogenesis of AD. Alternative mRNA splicing can generate isoforms of APP which contain a Kunitz protease inhibitor (KPI) domain. However, little is known about the physiological function of this domain. In the present study, the metabolic turnover of APP was examined in cultured chick sympathetic neurons. APP was labelled by incubating neurons for 5 h with [35S]methionine and [35S]cysteine. Intracellular labelled APP decayed in a biphasic pattern suggesting that trafficking occurs through two metabolic compartments. The half-lives for APP in each compartment were 1.5 and 5.7 h, respectively. A small fraction (10%) of the total APP was secreted into the culture medium where it was degraded with a half-life of 9 h. Studies using specific protease inhibitors demonstrated that this extracellular breakdown was due to cleavage by a trypsin-like serine protease that was secreted into the culture medium. Significantly, this protease was inhibited by a recombinant isoform of APP (sAPP751), which contains a region homologous to the Kunitz protease inhibitor (KPI) domain. These results suggest that KPI forms of APP regulate extracellular cleavage of secreted APP by inhibiting the activity of a secreted APP-degrading protease.

Alzheimer Disease↗

PubMed1999

mRNA degradation in bacteria.

Messenger RNAs in prokaryotes exhibit short half-lives when compared with eukaryotic mRNAs. Considerable progress has been made during recent years in our understanding of mRNA degradation in bacteria. Two major aspects determine the life span of a messenger in the bacterial cell. On the side of the substrate, the structural features of mRNA have a profound influence on the stability of the molecule. On the other hand, there is the degradative machinery. Progress in the biochemical characterization of proteins involved in mRNA degradation has made clear that RNA degradation is a highly organized cellular process in which several protein components, and not only nucleases, are involved. In Escherichia coli, these proteins are organized in a high molecular mass complex, the degradosome. The key enzyme for initial events in mRNA degradation and for the assembly of the degradosome is endoribonuclease E. We discuss the identified components of the degradosome and its mode of action. Since research in mRNA degradation suffers from dominance of E. coli-related observations we also look to other organisms to ask whether they could possibly follow the E. coli standard model.

Bacteria↗

PubMed1999

Thioredoxin is involved in oxygen-regulated formation of the photosynthetic apparatus of Rhodobacter sphaeroides.

Thioredoxin, a redox active protein, has been previously demonstrated to be essential for growth of the anoxygenic photosynthetic bacterium Rhodobacter sphaeroides. In the present study, the involvement of thioredoxin in the formation of the photosynthetic apparatus of R. sphaeroides WS8 was investigated by construction and analysis of a mutant strain disrupted for the chromosomal trxA copy and carrying a plasmid-borne copy of trxA under the control of the hybrid ptrc promoter inducible by IPTG (isopropyl-beta-D-thiogalactopyranoside). This strain was viable in the absence of IPTG but was affected in pigmentation. When shifted from high to low oxygen tension conditions, the trxA mutant showed a reduced bacteriochlorophyll content in comparison to that of the wild type. Although thioredoxin is able to regulate aminolevulinic acid (ALA) synthase (the first enzyme of the tetrapyrrole biosynthetic pathway) activity by a dithiol-disulfide exchange, our mutant strain exhibited a level of ALA synthase activity identical to that of the wild type, suggesting that thioredoxin is involved in other steps to regulate the synthesis of the photosynthetic apparatus. Accordingly, we showed that the trxA mutation affects the oxygen-regulated expression of the puf operon encoding the pigment-binding proteins of the light-harvesting and reaction center complexes. Upon transition from aerobic to semiaerobic growth conditions, the maximal puf mRNA level was found to be 40 to 50% lower in the mutant strain than in the wild type. The stability of the puf transcripts was identical in both strains grown under low oxygen tension, indicating that the role of thioredoxin in regulating puf expression occurs at the transcriptional level.

5-Aminolevulinate Synthetase↗

PubMed1999

RNase E enzymes from rhodobacter capsulatus and Escherichia coli differ in context- and sequence-dependent in vivo cleavage within the polycistronic puf mRNA.

The 5' pufQ mRNA segment and the pufLMX mRNA segment of Rhodobacter capsulatus exhibit different stabilities. Degradation of both mRNA segments is initiated by RNase E-mediated endonucleolytic cleavage. While Rhodobacter RNase E does not discriminate between the different sequences present around the cleavage sites within pufQ and pufL, Escherichia coli RNase E shows preference for the sequence harboring more A and U residues.

Bacterial Proteins↗

PubMed1998

Different cleavage specificities of RNases III from Rhodobacter capsulatus and Escherichia coli.

23S rRNA in Rhodobacter capsulatus shows endoribonuclease III (RNase III)-dependent fragmentation in vivo at a unique extra stem-loop extending from position 1271 to 1331. RNase III is a double strand (ds)-specific endoribonuclease. This substrate preference is mediated by a double-stranded RNA binding domain (dsRBD) within the protein. Although a certain degree of double strandedness is a prerequisite, the question arises what structural features exactly make this extra stem-loop an RNase III cleavage site, distinguishing it from the plethora of stem-loops in 23S rRNA? We used RNase III purified from R.capsulatus and Escherichia coli, respectively, together with well known substrates for E.coli RNase III and RNA substrates derived from the special cleavage site in R.capsulatus 23S rRNA to study the interaction between the Rhodobacter enzyme and the fragmentation site. Although both enzymes are very similar in their amino acid sequence, they exhibit significant differences in binding and cleavage of these in vitro substrates.

Bacterial Proteins↗