Proteolytic cleavage of the Chlamydia pneumoniae major outer membrane protein in the absence of Pmp10

The genome of the obligate intracellular bacteria Chlamydia pneumoniae contains 21 genes encoding polymorphic membrane proteins (Pmp). While no function has yet been attributed to the Pmps, they may be involved in an antigenic variation of the Chlamydia surface. It has previously been demonstrated that Pmp10 is differentially expressed in the C. pneumoniae CWL029 isolate. To evaluate whether the absence of Pmp10 in the outer membrane causes further changes to the C. pneumoniae protein profile, we subcloned the CWL029 isolate and selected a clone with minimal Pmp10 expression. Subsequently, we compared the proteome of the CWL029 isolate with the proteome of the subcloned strain and identified a specific cleavage of the C-terminal part of the major outer membrane protein (MOMP), which occurred only in the absence of Pmp10. In contrast, when Pmp10 was expressed we predominantly observed full-length MOMP. No other proteins appeared to be regulated according to the presence or absence of Pmp10. These results suggest a close association between MOMP and Pmp10, where Pmp10 may protect the C-terminal part of MOMP from proteolytic cleavage.     

Optimal fed batch experiment design for estimation of monod kinetics of Azospirillum brasilense: from theory to practice

In this paper the problem of reliable and accurate parameter estimation for unstructured models is considered. It is illustrated how a theoretically optimal design can be successfully translated into a practically feasible, robust, and informative experiment. The well-known parameter estimation problem of Monod kinetic parameters is used as a vehicle to illustrate our approach. As known for a long time, noisy batch measurements do not allow for unique and accurate estimation of the kinetic parameters of the Monod model. Techniques of optimal experiment design are, therefore, exploited to design informative experiments and to improve the parameter estimation accuracy. During the design process, practical feasibility has to be kept in mind. The designed experiments are easy to implement in practice and do not require additional monitoring equipment. Both design and experimental validation of informative fed batch experiments are illustrated with a case study, namely, the growth of the nitrogen-fixing bacteria Azospirillum brasilense.     

Hi-C deconvolution of a textile dye–related microbiome reveals novel taxonomic landscapes and links phenotypic potential to individual genomes

International Microbiology - Microbial biodiversity is represented by a variety of genomic landscapes adapted to dissimilar environments on Earth. These genomic landscapes contain functional...     

The Barnes Maze Task Reveals Specific Impairment of Spatial Learning Strategy in the Intrahippocampal Kainic Acid Model for Temporal Lobe Epilepsy

Neurochemical Research - Temporal lobe epilepsy (TLE) is an acquired form of focal epilepsy, in which patients not only suffer from unprovoked, devastating seizures, but also from severe...     

Constitutive signaling of the human cytomegalovirus-encoded chemokine receptor US28

Previously it was shown that the HHV-8-encoded chemokine receptor ORF74 shows considerable agonist-independent, constitutive activity giving rise to oncogenic transformation (Arvanitakis, L., Geras-Raaka, E., Varma, A., Gershengorn, M. C., and Cesarman, E. (1997) Nature 385, 347-350). In this study we report that a second viral-encoded chemokine receptor, the human cytomegalovirus-encoded US28, also efficiently signals in an agonist-independent manner. Transient expression of US28 in COS-7 cells leads to the constitutive activation of phospholipase C and NF-kappaB signaling via G(q/11) protein-dependent pathways. Whereas phospholipase C activation is mediated via Galpha(q/11) subunits, the activation of NF-kappaB strongly depends on betagamma subunits with a preference for the beta(2)gamma(1) dimer. The CC chemokines RANTES (regulated on activation, normal T cell expressed and secreted) and MCP-1 (monocyte chemotactic protein-1) act as neutral antagonists at US28, whereas the CX(3)C chemokine fractalkine acts as a partial inverse agonist with IC(50) values of 1-5 nm. Our data suggest that a high level of constitutive activity might be a more general characteristic of viral G protein-coupled receptors and that human cytomegalovirus might exploit this G protein-coupled receptor property to modulate the homeostasis of infected cells via the early gene product US28.     

Untersuchungen über die Osteon- und Lamellenformen im Extremitätenskelet des Erwachsenen

Zusammenfassung Auf Grund der Untersuchung von Knochenquerschnitten eines gesunden 43jährigen Mannes, bestätigt durch noch zu veröffentlichende Untersuchungen an Tibien verschiedener Individuen, werden die Strukturformen des Knochens nach ihrem Querschnittsbild beschrieben.Es wird zwischen Osteonen und Tangentiallamellen unterschieden. Zu den Tangentiallamellen mit dem mehr oder minder parallelen Verlauf zur Knochenoberfläche gehören die bisher als Generallamellen und Schaltlamellen beschriebenen Systeme.Soweit eine Lamellengliederung vorhanden ist, zeichnen sich die Tangentiallamellen durch den strengen Wechsel zwischen flach und steil gewickelten aus.Auf Grund des Querschnittsbildes werden verschiedene Osteonformen unterschieden. Die Größe des einzelnen Osteonquerschnittes wird mit Hilfe der Lamellenzahl bestimmt. Gleichzeitig wird die Steigungsfolge beachtet, d. h. der Wechsel des Kollagenfaserverlaufs von Lamelle zu Lamelle.Es ergibt sich, daß die kleineren Osteone überwiegend in der peripheren Schnitthälfte, die größeren dagegen in der zentralen liegen. Der regelmäßige Wechsel der Steigungsfolge nimmt von den kleineren zu den größeren Osteonen hin ab, die mehr steile Verlaufsweise dagegen zu. Die kleineren Größenklassen lassen häufiger die lamelläre Gliederung vermissen als die großen.Abschließend wird erörtert, daß sowohl das Osteon wie die Lamelle nur als eine besondere Lagerungsform der Kollagenfasern im Knochen angesehen werden können. Der Begriff Osteon wurde in Anlehnung an die Begriffe der überwiegend zellulären Einheiten Neuron und Chondron bzw. der sog. Entwicklungs- und Funktionseinheit Nephron gebildet. Die zirkuläre Lagerung der Kollagenfasern hat aller Voraussicht nach eine besondere festigkeitstheoretische Bedeutung. Sie ist aber abhängig vom Gefäßbaum, an dessen Verzweigungen Doppelbildungen auftreten. Diese Doppelbildungen teilen sich und begleiten die Gefäßäste. Sie werden damit zum Osteon, das sich nach Querschnittsgröße und Wicklung in benachbarten Querschnitten verschieden verhalten kann. Die zirkuläre Wicklung führt nicht zu individuellen Gebilden, die den Knochen wie Bausteine aufbauen. Sie stellt ein System dar, das den Gefäßbaum im Knochen in mehr oder minder kontinuierlichem Zusammenhang begleitet. Die zirkulären Wicklungen gehen ohne Abgrenzung in die übrigen Lamellensysteme, die Tangentiallamellen, über. Osteone und Tangentiallamellen erscheinen damit als eine übergeordnete Lagerungsform der Kollagenfasern. Die nächstniedere Stufe der Lagerung ist die Lamelle.Mit Unterstützung der Deutschen Forschungsgemeinschaft.     

Quantitative cleavage of the N-glycosidic bond under the normal conditions of methanolysis used for the analysis of glycoprotein monosaccharides

The most common method used for the liberation of monosaccharides from glycoprotein N-glycans involves anhydrous methanolysis because it liberates almost quantitatively monosaccharides as O-methylglycosides, which are resistant to further degradation. However, it is generally assumed that this method does not cleave quantitatively the N-glycosidic bonds. This paper demonstrates that classical methanolysis conditions quantitatively cleave the N-glycosidic bond (96%), liberating glucosamine (and not its O-methylglycosides) and other minor reaction products which were identified. Because other N-acetyl-d-glucosamine (GlcNAc) residues are quantitatively liberated as the O-methylglycosides of glucosamine, the GlcNAc residue involved in the N-glycosidic bond is separated from the others using gas chromatography of heptafluorobutyrate derivatives.     

Untreated silicone breast implant rupture

Implant rupture is a well-known complication of breast implant surgery that can pass unnoticed by both patient and physician. To date, no prospective study has addressed the possible health implications of silicone breast implant rupture. The aim of the present study was to evaluate whether untreated ruptures are associated with changes over time in magnetic resonance imaging findings, serologic markers, or self-reported breast symptoms. A baseline magnetic resonance imaging examination was performed in 1999 on 271 women who were randomly chosen from a larger cohort of women having cosmetic breast implants for a median period of 12 years (range, 3 to 25 years). A follow-up magnetic resonance imaging examination was carried out in 2001, excluding women who underwent explantation in the period between the two magnetic resonance imaging examinations (n = 44). On the basis of these examinations, the authors identified 64 women who had at least one ruptured implant at the first magnetic resonance imaging examination and, for comparison, all women who had intact implants at both examinations (n = 98). Magnetic resonance images from the two examinations were compared and changes in rupture configuration were evaluated. Comparisons were also made for self-reported breast symptoms occurring during the study period and for changes in serum values of antinuclear antibodies, rheumatoid factor, and cardiolipin antibodies immunoglobulin G and immunoglobulin M. The majority of the women with implant rupture had no visible magnetic resonance imaging changes of their ruptured implants. For 11 implants (11 percent) in 10 women, the authors observed progression of silicone seepage, either as a conversion from intracapsular into extracapsular rupture (n = 7), as progression of extra-capsular silicone (n = 3), or as increasing herniation of the silicone within the fibrous capsule (n = 1); however, in most cases, these changes were minor. Some changes could be ascribed to trauma, but others seemed spontaneous. There was no increase in levels of autoantibodies during the study period in either study group. Women with untreated implant ruptures reported a significant increase in nonspecific breast changes (odds ratio, 2.1; 95 percent confidence interval, 1.2 to 3.8) compared with women without ruptures. On the basis of this first study of women with untreated silicone breast implant rupture, the authors conclude that implant rupture is a relatively harmless condition, which only rarely progresses and gives rise to notable symptoms. Even so, because of a small risk of silicone spread, the authors suggest that women with implant ruptures be followed clinically, if not operated on. Because implant ruptures often occur asymptomatically, any woman with silicone implants, regardless of rupture status, should be evaluated at regular intervals.     

Glomulin is predominantly expressed in vascular smooth muscle cells in the embryonic and adult mouse

Mutations in the glomulin gene result in dominantly inherited vascular lesions of the skin known as glomuvenous malformations (GVMs). These lesions are histologically distinguished by their distended vein-like channels containing characteristic 'glomus cells', which appear to be incompletely or improperly differentiated vascular smooth muscle cells (VSMCs). The function of glomulin is currently unknown. We studied glomulin expression during murine development (E9.5 days post-coitum until adulthood) by non-radioactive in situ hybridization. Glomulin was first detected at E10.5 dpc in cardiac outflow tracts. Later, it showed strong expression in VSMCs as well as a limited expression in the perichondrium. At E11.5-14.5 dpc glomulin RNA was most abundant in the walls of the large vessels. At E16.5 dpc expression was also detectable in smaller arteries and veins. The high expression of glomulin in murine vasculature suggests an important role for glomulin in blood vessel development and/or maintenance, which is supported by the vascular phenotype seen in GVM patients with mutations in this gene.