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Biomedical subjects

C Marschall

Publications and source records attributed to C Marschall.

9 recordsLinked to original sources

UVB increases urokinase-type plasminogen activator receptor (uPAR) expression.

Keratinocytes synthesize and secrete urokinase-type plasminogen activator, which binds to its specific receptor on keratinocytes. When bound to urokinase-type plasminogen activator receptor, urokinase-type plasminogen activator proteolytically converts surface bound plasminogen to plasmin, which in turn cleaves many extracellular components leading to pericellular proteolysis. The activation of the urokinase system has been observed during re-epithelialization of skin wounds and in lesions of the autoimmune blistering skin disease pemphigus. As pemphigus is photoinducible, we investigated the effect of ultraviolet B on urokinase-type plasminogen activator and urokinase-type plasminogen activator receptor expression in the epidermal keratinocyte cell line A431. Ultraviolet B increased cellular and secreted urokinase-type plasminogen activator protein (enzyme-linked immunosorbent assay) and urokinase-type plasminogen activator receptor cell surface expression (flow cytometry) 24 h postirradiation. Northern blot analysis indicated that ultraviolet B increased urokinase-type plasminogen activator receptor mRNA. Compared with a more rapid mRNA induction by epidermal growth factor (maximal after 4 h) the ultraviolet B response was maximal after 24 h and prolonged up to 36 h. The mRNA induction was not dependent on protein synthesis as judged by cycloheximide incubation. Ultraviolet B did not influence urokinase-type plasminogen activator receptor mRNA stability (actinomycin D incubation). A transiently transfected chloramphenicol acetyltransferase-reporter construct containing a -398/+51 urokinase-type plasminogen activator receptor promoter fragment was activated when cells were exposed to ultraviolet B. This induction was almost completely abolished by mutating a -182/-176 AP-1 binding sequence. Ultraviolet B increased the binding capacity at this AP-1 motif in electrophoretic mobility shift assays. These data identify a distinct transcriptional mechanism by which ultraviolet B induces urokinase-type plasminogen activator receptor. The epidermal induction of components of the proteolytic urokinase system by ultraviolet B may help explain the photoinducibility of pemphigus lesions.

Binding Sites↗

Molecular analysis of the regulation of csiD, a carbon starvation-inducible gene in Escherichia coli that is exclusively dependent on sigma s and requires activation by cAMP-CRP.

The general stress-induced sigma subunit sigma s of Escherichia coli RNA polymerase is closely related to the vegetative sigma factor sigma 70. In view of their very similar promoter specificity in vitro, it is unclear how sigma factor selectivity in the expression of sigma s-dependent genes is generated in vivo. The csiD gene is such a strongly sigma s-dependent gene. In contrast to sigma s, which is induced in response to many different stresses, csiD, whose expression is driven from a single promoter, is induced by carbon starvation only. To our knowledge, the csiD promoter is the first characterized promoter which is not only exclusively dependent on sigma s-containing RNA polymerase (E sigma s), but also requires an activator, cAMP-CRP. In addition, leucine-responsive regulatory protein (Lrp) acts as a positive modulator of csiD expression. Also in vitro, E sigma s is more efficient than E sigma 70 in csiD promoter binding, open complex formation and run-off transcription, which might be due to the poor match of the csiD -35 region to the sigma 70 consensus and to transcription by E sigma s being less dependent on contacts in this region. By DNase I protection experiments, a cAMP-CRP binding site centered at -68.5 nucleotides upstream of the csiD transcriptional start site was identified. While cAMP-CRP stimulates E sigma 70 binding, it does not promote open complex formation by E sigma 70, but does so in conjunction with E sigma s. With linear templates, cAMP-CRP significantly stimulates E sigma s-mediated in vitro transcription, whereas transcription by E sigma 70 is negligible and hardly stimulated by cAMP-CRP. These findings may reflect different or less stringent positional requirements for an activator site for E sigma s than for E sigma 70, and indicate that cAMP-CRP contributes to sigma factor selectivity at the csiD promoter. In vitro transcription experiments with super-coiled templates, however, revealed significant cAMP-CRP-stimulated transcription also by E sigma 70. Yet, under these conditions, H-NS was found to restore E sigma s specificity by strongly interfering with cAMP-CRP/E sigma 70-dependent transcription. Lrp strongly and cooperatively binds to multiple sites located between positions -14 and -102 (in a way that suggests DNA wrapping around multiple Lrp molecules) and moderately stimulates in vitro transcription, especially with E sigma s. In summary, we conclude that the csiD promoter has an intrinsic preference for E sigma s, but that also protein factors such as cAMP-CRP, Lrp and probably H-NS as well as DNA conformation contribute to its strong E sigma s selectivity. Furthermore, this strong E sigma s preference in combination with a requirement for high concentrations of the essential activator cAMP-CRP ensures csiD expression under conditions of carbon starvation, but not other stress conditions.

Bacterial Proteins↗

Heat shock regulation of sigmaS turnover: a role for DnaK and relationship between stress responses mediated by sigmaS and sigma32 in Escherichia coli.

The cellular level of the rpoS-encoded sigmaS subunit of RNA polymerase increases in response to various stress situations that include starvation, high osmolarity, and shift to acid pH, and these different stress signals differentially affect rpoS translation and/or sigmaS stability. Here we demonstrate that sigmaS is also induced by heat shock and that this induction is exclusively due to an interference with sigmaS turnover. Some sigmaS-dependent genes exhibit similar heat shock induction, whereas others are not induced probably because they need additional regulatory factors that might not be present under conditions of heat shock or exponential growth. Despite its induction, sigmaS does not seem to contribute to heat adaptation but may induce cross-protection against different stresses. While sigmaS is not involved in the regulation of the heat shock sigma factor sigma32, the heat shock protein DnaK has a positive role in the posttranscriptional control of sigmaS. The present evidence suggests that DnaK is involved in the transduction of two of the signals that result in reduced sigmaS turnover, i.e., heat shock and carbon starvation. Heat shock induction of sigmaS also clearly indicates that a cessation of growth or even a reduction of the growth rate is not a prerequisite for the induction of sigmaS and sigmaS-dependent genes and underscores the importance of sigmaS as a general stress sigma factor.

Bacterial Proteins↗

Regulatory characteristics and promoter analysis of csiE, a stationary phase-inducible gene under the control of sigma S and the cAMP-CRP complex in Escherichia coli.

Sigma-S and the cAMP-CRP complex are global regulatory factors involved in stationary-phase induction of large groups of genes in Escherichia coli. csiE, a gene located at 57.25 min (co-ordinate 2674) of the physical map of the E. coli chromosome, is under the control of both of these factors. Sigma-S plays a positive, though not absolutely essential, role in the expression of csiE. Regulation by cAMP-CRP has both positive and negative elements, with the latter being dependent on the presence of sigma s, whose expression is negatively influenced by cAMP-CRP. csiE has a single transcriptional start site located 33 bp upstream of the initiation codon. By a 5'-deletion approach, we show that 72 bp upstream of the csiE transcriptional start site are sufficient for regulation by sigma s and cAMP-CRP. A deletion upstream of nucleotide -38 with respect to the start site eliminates positive cAMP-CRP control and makes the remaining expression fully dependent on sigma S. Our results indicate that transcription at the csiE promoter can be initiated in vivo by sigma S-containing RNA polymerase alone as well as by sigma 70-containing RNA polymerase in conjunction with cAMP-CRP or a cAMP-CRP-dependent secondary regulator. The promoter region of poxB, the structural gene for pyruvate oxidase, which is also under the control of sigma S and cAMP-CRP, is very similar to the corresponding region of csiE, suggesting a similar regulatory mechanism also for poxB.

Bacterial Proteins↗

Role for the histone-like protein H-NS in growth phase-dependent and osmotic regulation of sigma S and many sigma S-dependent genes in Escherichia coli.

The sigma S subunit of RNA polymerase (encoded by the rpoS gene) is the master regulator in a complex regulatory network that controls stationary-phase induction and osmotic regulation of many genes in Escherichia coli. Here we demonstrate that the histone-like protein H-NS is also a component of this network, in which it functions as a global inhibitor of gene expression during the exponential phase of growth. On two-dimensional gels, at least 22 sigma S-controlled proteins show increased expression in an hns mutant. H-NS also inhibits the expression of sigma S itself by a mechanism that acts at the posttranscriptional level. Our results indicate that relief of repression by H-NS plays a role in stationary-phase induction as well as in hyperosmotic induction of rpoS translation. Whereas certain sigma S-dependent genes (e.g., osmY) are only indirectly regulated by H-NS via its role in the control of sigma S expression, others are also H-NS-regulated in a sigma S-independent manner. (For this latter class of genes, rpoS hns double mutants show higher levels of expression than mutants deficient in rpoS alone.) In addition, we demonstrate that the slow-growth phenotype of hns mutants is suppressed in hns rpoS double mutants and that many second-site suppressor mutants that spontaneously arise from hns strains carry lesions that affect the expression of sigma S.

Bacterial Outer Membrane Proteins↗

Hypothalamic monoamines in lizards (Lacerta). A histofluorescence study.

The morphological organization of monoaminergic cells and fibers in the hypothalamus of the lizards Lacerta sicula and Lacerta muralis was investigated by fluorescence histochemistry. An extensive monoaminergic system emanates from the nucleus organi paraventricularis (NOP), a circumventricular organ of the medial and posterior hypothalamus containing numerous monoaminergic perikarya. Fluorescent processes extending from these cells end as intraventricular thickenings. An extensive fiber system presumably arising from the NOP innervates the accompanying nucleus ventromedialis hypothalami (NVH) as well as the nucleus periventricularis hypothalami (NPH) and the median eminence. A monoaminergic fiber path of extrahypothalamic origin enters the preoptic/anterior hypothalamus, terminating in the nucleus paraventricularis (NP). A discrete pathway of catecholaminergic fibers courses through the hypothalamus along the ventral border of the optic tract. Levels of fluorescence intensity are highest in the spring and in castrated animals and lowest in lizards during testicular regression.

Amines↗

[Cortical strut grafts, an alternative to conventional plating in periprosthetic fractures of the femur].

AIM: In periprosthetic fracture therapy plating has a failure rate between 30 to 50 %. Cortical allografts proved to unite constantly in revision surgery of the femoral shaft. The hypotheses to be answered in this study was whether the use of cortical allografts increases the success rate in contrast to plating. METHOD: A retrospective case control study has been done analysing the results in periprosthetic fractures of the femur around or below the tip of a stable femoral component (Vancouver B1, Mont III + IV). RESULTS: In 12 plating cases an average of 2.67 units of blood was transfused (0 - 6). In the follow-up two refractures, one loosening of the femoral component and one varus deformity greater than 10 degrees occurred. In 6 patients a total of 9 reoperations had to be done. According to the classification of Mont the results were excellent in 50 %, good in 16.7 % and poor in 33.3 %. 7 patients with strut grafts received an average of 1,43 units of blood transfused (0 - 3). In one patient with a 16 degrees varus deformity of the femur, due to multiple previous operations, a deformity of 21 degrees had to be accepted. Until now, no further operation had to be done in this group. The results were classified as excellent in 5 cases (71.4 %) and good in the other 2 (28,6 %). There was no poor result. The difference between the groups was statistically significant in the number of reoperations (p < 0. 05). CONCLUSION: In this study, with a low number so far, the strut graft group reached better results in every single variable, with a significant reduction of reoperations.

Aged↗

[Osteogenesis imperfecta Type 1: a case presentation with a new mutation in gene COL1A1].

In a 4 year old girl the diagnosis osteogenesis imperfecta type I was suspected by following clinical criteria: four fractures after small trauma, intensive blue sclera, anomalies of dental enamel, macrocephalie with frontal bassing. Clinical diagnosis could be verified by moleculargenetic analysis, a newly recognized heterozygous point mutation (Arg420Stop) in the COL1A1-gene was found.

Ankle Injuries↗