Neutrophil Gelatinase-Associated Lipocalin (NGAL) is Associated with Symptomatic Carotid Atherosclerosis and Drives Pro-inflammatory State In Vitro.

OBJECTIVE: Neutrophil gelatinase-associated lipocalin (NGAL), a protein found in activated neutrophils, is expressed in kidney tubule cells in response to noxious stimuli, and is thus recognized as a marker of acute kidney injury. Recent studies have suggested that NGAL could also have pathophysiological importance in cardiovascular diseases. The aim of the present study was to examine NGAL expression in human carotid endarterectomy tissues ex vivo as well as the effects of NGAL in the main cell types involved in atherogenesis, namely in human macrophages, endothelial cells, and smooth muscle cells in vitro. METHODS: NGAL protein was analyzed in human endarterectomy samples from patients with asymptomatic and symptomatic carotid stenosis by immunofluorescence, and NGAL mRNA expression was detected using RealTime-PCR. Human monocyte derived macrophages (MDM), human coronary artery smooth muscle cells (HCASMC), and human umbilical vein endothelial cells (HUVEC) were treated with recombinant human (rh) NGAL at different concentrations. Interleukin (IL)-6, IL-8, and monocyte chemo-attractant protein-1 (MCP-1) were determined by specific enzyme linked immunosorbent assays (ELISAs) in culture supernatants of such treated cells. RESULTS: Expression of NGAL protein was demonstrated by macrophages, smooth muscle cells, and endothelial cells in human carotid atherosclerotic tissue. NGAL mRNA expression was detected at a higher rate in atherosclerotic tissue of patients with symptomatic carotid stenosis (in 70%; n = 19) compared with asymptomatic patients (in 37%; n = 20, p < .001). Treatment of MDM, HCASMC, and HUVEC with rhNGAL led to a significant (p < 0.05) and concentration dependent increase of pro-inflammatory cytokines IL-6, IL-8, and MCP-1 in all cell types analyzed. CONCLUSION: By induction of pro-inflammatory mediators in human macrophages, smooth muscle cells and endothelial cells, NGAL, which is predominantly expressed in atherosclerotic plaques of symptomatic patients, could be involved in creating the local and systemic pro-inflammatory environment characteristic for atherosclerosis.

Characterization of patients at high risk of melanoma in Austria.

BACKGROUND: Risk of melanoma is determined by genetic and exogenous factors. Only a few studies have included both characteristics in a comprehensive multivariable analysis. OBJECTIVES: To find determinants of patients at high risk of melanoma in Austria, including phenotype, genotype and lifestyle characteristics in comprehensive analyses. METHODS: In total, 1668 patients with melanoma from the M3 case-control study were studied. Overall, 567 participants were sequenced for CDKN2A, 232 for CDK4, 123 for MITF encoding the variant E318K and 964 for MC1R. RESULTS: Patients with melanoma with a positive family history (n = 190, 11·6%), multiple primary melanomas (n = 261, 15·7%) and younger age (< 50 years, n = 675, 40·5%) were defined as being at high risk. All other patients with melanoma were defined as the reference group. We found significant differences between those two groups and between the high-risk subgroups (positive family history, multiple primary melanomas and younger age). Pigmentation phenotype was associated with the high-risk group in general (childhood freckling, odds ratio 1·46, P = 0·007; blond/reddish hair colour, odds ratio 1·43, P = 0·011). Patients with a positive family history and patients with early-onset disease were similar regarding both their phenotypic characteristics and external factors. Established high-risk mutations in CDKN2A were found in cases with a positive family history (n = 12) or multiple melanomas (n = 2). Moreover, we found three patients carrying the MITF p.E318K variant, two with a CDK4 variant and seven with nonsynonymous MC1R variants with undescribed biological significance, of which four were predicted as damaging. CONCLUSIONS: Austrian patients could represent a reservoir for novel genetic variants. Further investigation of populations in Central and Eastern Europe might reveal more novel and disease-relevant variants.

Interleukin-33 induces urokinase in human endothelial cells--possible impact on angiogenesis.

BACKGROUND: Urokinase-type plasminogen activator (u-PA) plays a pivotal role in extracellular proteolysis and is thought to be critically involved in the modulation of angiogenesis. Interleukin (IL)-33 is a member of the IL-1 cytokine family, which is thought to act as danger signal that is released from cells after injury. IL-33 is involved in the pathogenesis of various inflammatory diseases and previously was shown to induce angiogenesis and inflammatory activation of endothelial cells. OBJECTIVE: We investigated the impact of IL-33 on u-PA in endothelial cells as a new possible function for IL-33. METHODS AND RESULTS: We could demonstrate that IL-33 upregulated u-PA mRNA expression and protein production in human coronary artery and human umbilical vein endothelial cells in a time- and concentration-dependent manner via interaction with its receptor ST2 and activation of the nuclear factor-κB pathway but independent of autocrine IL-1-induced effects. The hydroxymethylglutaryl-coenzyme A reductase inhibitor simvastatin abrogated the IL-33-induced increase in u-PA, thus providing further evidence for pleiotropic effects of statins. IL-33 induced u-PA-dependent capillary-like tube formation and vessel sprouting. In human carotid atherosclerotic plaques (n = 16), u-PA mRNA positively correlated with IL-33 mRNA expression (r = 0.780, P < 0.001). Furthermore, IL-33 and u-PA protein were detected in endothelial cells in these samples using fluorescence immunohistochemistry. CONCLUSIONS: We hypothesize that IL-33, representing a danger signal that is released after tissue damage, in addition to its role in the inflammatory activation of endothelial cells, is involved in u-PA-driven angiogenesis, a process that has been shown before to be linked to inflammation in various pathologies.

Actinic damage on the back is significantly determined by MC1R variants and previous sun exposure compared with other body sites in a multivariate analysis.

BACKGROUND: Only recently, site-dependent associations of actinic damage with melanoma were identified in our study population. OBJECTIVES: To elucidate the diverse aetiologies for actinic damage at different body sites. METHODS: We performed multivariate logistic regression analyses to identify independent risk factors for actinic damage on the face, hands and the back in 2112 participants of central European origin. RESULTS: For actinic damage on the face, age was the only risk factor that remained consistently significant in a multivariate analysis, whereas actinic damage on the back was predominantly associated with number of sunburns, freckles in childhood, holiday weeks and male sex. Moreover, we identified a particular significance of MC1R variants and dorsal actinic skin damage. CONCLUSIONS: The particular effect of MC1R variants and sun exposure during recreational time on dorsal actinic damage indicates that actinic damage on the back is more informative regarding susceptibility to sunlight and past sun exposure associated with melanoma risk.

[Enhanced automatic collision notification system. First field experience]. / Automatische Unfallmelder. Erste Erfahrungen aus der Anwendung.

Motor vehicle accidents are the leading cause of severe injuries. Outcome of these victims is substantially influenced by rescue time and primary trauma centre care. Nowadays, automatic crash notification systems (ACN) are available offering the major possibility of immediate initiation of the rescue chain and thereby shortening rescue time up to 50%. According to the EU commission a nationwide equipping of motor vehicles with automatic crash notification systems could save up to 2,500 lives per year. Therefore, all new vehicles should be equipped with an automatic crash notification system by 2014. BMW is the first manufacturer to install an enhanced automatic crash notification system (eACN), which could optimize the rescue triage by estimating the risk for severe injuries (MAIS3+) based on telemetric transmitted accident data.

Conserved RNA secondary structures in Picornaviridae genomes.

The family Picornaviridae contains important pathogens including, for example, hepatitis A virus and foot-and-mouth disease virus. The genome of these viruses is a single messenger-active (+)-RNA of 7200-8500 nt. Besides coding for the viral proteins, it also contains functionally important RNA secondary structures, among them an internal ribosomal entry site (IRES) region towards the 5'-end. This contribution provides a comprehensive computational survey of the complete genomic RNAs and a detailed comparative analysis of the conserved structural elements in seven of the currently nine genera in the family PICORNAVIRIDAE: Compared with previous studies we find: (i) that only smaller sections of the IRES region than previously reported are conserved at single base-pair resolution and (ii) that there is a number of significant structural elements in the coding region. Furthermore, we identify potential cis-acting replication elements in four genera where this feature has not been reported so far.

Comparison of real world side impact/rollover collisions with and without thorax airbag/head protection system: a first field experience study.

After the introduction of the Thorax Airbag (TA) and the Head Protection System (HPS) by BMW there has been a significant reduction of injuries in real-world collisions. Comparison of similar collisions (in-depth collision analyses) of vehicles with and without HPS/TA indicates that the effectiveness of the system was credible. Minor injuries (AIS 1) increase while serious injuries (AIS 3+) are reduced. Based on the limited cases available, a proper statistical sampling could not be achieved at this time, however the results are to be understood as indicative of a trend.

Automatic detection of conserved RNA structure elements in complete RNA virus genomes.

We propose a new method for detecting conserved RNA secondary structures in a family of related RNA sequences. Our method is based on a combination of thermodynamic structure prediction and phylogenetic comparison. In contrast to purely phylogenetic methods, our algorithm can be used for small data sets of approximately 10 sequences, efficiently exploiting the information contained in the sequence variability. The procedure constructs a prediction only for those parts of sequences that are consistent with a single conserved structure. Our implementation produces reasonable consensus structures without user interference. As an example we have analysed the complete HIV-1 and hepatitis C virus (HCV) genomes as well as the small segment of hantavirus. Our method confirms the known structures in HIV-1 and predicts previously unknown conserved RNA secondary structures in HCV.

Spontaneous and engineered deletions in the 3' noncoding region of tick-borne encephalitis virus: construction of highly attenuated mutants of a flavivirus.

The flavivirus genome is a positive-strand RNA molecule containing a single long open reading frame flanked by noncoding regions (NCR) that mediate crucial processes of the viral life cycle. The 3' NCR of tick-borne encephalitis (TBE) virus can be divided into a variable region that is highly heterogeneous in length among strains of TBE virus and in certain cases includes an internal poly(A) tract and a 3'-terminal conserved core element that is believed to fold as a whole into a well-defined secondary structure. We have now investigated the genetic stability of the TBE virus 3' NCR and its influence on viral growth properties and virulence. We observed spontaneous deletions in the variable region during growth of TBE virus in cell culture and in mice. These deletions varied in size and location but always included the internal poly(A) element of the TBE virus 3' NCR and never extended into the conserved 3'-terminal core element. Subsequently, we constructed specific deletion mutants by using infectious cDNA clones with the entire variable region and increasing segments of the core element removed. A virus mutant lacking the entire variable region was indistinguishable from wild-type virus with respect to cell culture growth properties and virulence in the mouse model. In contrast, even small extensions of the deletion into the core element led to significant biological effects. Deletions extending to nucleotides 10826, 10847, and 10870 caused distinct attenuation in mice without measurable reduction of cell culture growth properties, which, however, were significantly restricted when the deletion was extended to nucleotide 10919. An even larger deletion (to nucleotide 10994) abolished viral viability. In spite of their high degree of attenuation, these mutants efficiently induced protective immune responses even at low inoculation doses. Thus, 3'-NCR deletions represent a useful technique for achieving stable attenuation of flaviviruses that can be included in the rational design of novel flavivirus live vaccines.

Secondary structure of the 3'-noncoding region of flavivirus genomes: comparative analysis of base pairing probabilities.

The prediction of the complete matrix of base pairing probabilities was applied to the 3' noncoding region (NCR) of flavivirus genomes. This approach identifies not only well-defined secondary structure elements, but also regions of high structural flexibility. Flaviviruses, many of which are important human pathogens, have a common genomic organization, but exhibit a significant degree of RNA sequence diversity in the functionally important 3'-NCR. We demonstrate the presence of secondary structures shared by all flaviviruses, as well as structural features that are characteristic for groups of viruses within the genus reflecting the established classification scheme. The significance of most of the predicted structures is corroborated by compensatory mutations. The availability of infectious clones for several flaviviruses will allow the assessment of these structural elements in processes of the viral life cycle, such as replication and assembly.

Sex pheromone of European corn borer. : Ostrinia nubilalis: Polymorphism in various laboratory and field strains.

Sex gland extracts ofOstrinia nubilalis females collected in the wild or laboratory-reared from Switzerland, Italy, and Hungary were analyzed. Individuals collected in the north of Switzerland contained (Z)- and (E)-11-tetradecenyl acetate at the approximate ratio of 97∶3 (Z type), in accordance with field responses of males and previous findings. On the other hand, females from a laboratory culture derived from field collections made in the same area and reared for four to five generations contained theZ andE isomers at ratios of ca. 3∶97 and 35∶65, respectively. In the south, one of the eight wild females analyzed was of theZ type and the rest intermediate, whereas males were predominantly trapped with blends of the two isomers containing 60 to 97E. In a laboratory culture reared for one to two generations from corn borers collected in Hungary, three of nine females were of the intermediate type and the restZ. Small amounts of (Z)-11-hexadecenyl acetate were detected in female glands of theE strain; however, no effect of this compound could be observed in the field.

Sex pheromone ofEupoecilia ambiguella female: Analysis and male response to ternary blend.

Sex gland extracts and washes ofEupoecilia ambiguella contain 10-20 ng/female of the primary sex pheromone componentZ-9-dodecenyl acetate (Z9-12∶Ac), accompanied by a number of related compounds. These areE-9-dodecenyl acetate (E9-12∶Ac),Z-9-dodecen-1-ol (Z9-12∶OH), saturated acetates of 12, 16, 18, and 20 carbons, and traces of a doubly unsaturated acetate, tentatively identified as a 9,11-dodecadienyl acetate. Octadecyl acetate predominates among the pheromone-related components, making up 1-2, occasionally 20-30 times the amount ofZ9-12∶Ac. The same compounds were also found in field-collected females and in effluvia.Z-9-Undecenyl acetate, which is a male attractant on its own, was also found in a sample of female effluvia. A hierarchy is observed in the ethological function of the pheromone components.Z9-12∶Ac is an attractant forE. ambiguella males. Dodecyl acetate (12∶Ac) is not attractive on its own but augments male catch when added to the main attractant. Addition of 18∶Ac augments attraction only when bothZ9-12∶Ac and 12∶Ac are present. Windtunnel tests demonstrate that 18∶Ac also raises the disorientation threshold, as previously shown for 12∶Ac. Other compounds, with the possible exception of additional saturated acetates, had either no effect on trap catch or, in the case ofE9-12∶Ac,Z9-12∶OH, andE-9,11-dodecadienyl acetate, were inhibitory above a certain level. A blend of roughly equal parts ofZ9-12∶Ac, 12∶Ac, and 18∶Ac provides the best attractant blend forE. ambiguella known to date.

Effects of dodecyl acetate andZ-10-tridecenyl acetate on attraction ofEupoecilia ambiguella males to the main sex pheromone component,Z-9-Dodecenyl acetate.

Attraction ofE. ambiguella males toZ-9-dodecenyl acetate (Z9-12∶Ac), alone and in combination with dodecyl acetate (12∶Ac) orZ-10-tridecenyl acetate (Z10-13∶Ac) was tested in both wind-tunnel and field experiments. In the laboratory, response toZ9-12∶Ac reached a distinct dosage optimum at which attraction was nearly as good as to live females. Addition of 12∶Ac, a minor component of female glands and effluvia, had no effect at low doses ofZ9-12∶Ac and only marginally improved attraction at the optimum. However, inclusion of 12∶Ac with an overdose ofZ9-12∶ Ac fully restored activity. In the field,Z9-12∶Ac alone was not very attractive at any dose; catches markedly increased by adding 12∶Ac and climbed with increasing dose of each compound up to 10 mg. Optimum ratio ofZ9-12∶Ac to 12∶Ac was 1∶1 to 1∶5. Another compound,Z10-13∶Ac, had the same effect as 12∶Ac in the laboratory and the field but was effective at a lower dose. In the field, inclusion of the synergist permits use of high cap loads to attract more males, perhaps from greater distances.

Field attraction ofAgrotis segetum males in four european countries to mixtures containing three homologous acetates.

(Z)-5-Decenyl, (Z)-7-dodecenyl, and (Z)-9-tetradecenyl acetate were tested as single compounds and blends for field attraction to the turnip moth in Denmark, France, Hungary, and Switzerland. Best attraction was obtained when all three components were present. Attractants for other noctuids are described, e.g.,Meristis trigrammica, Hoplodrina alsines, Erastria trabealis, Euxoa nigricans, Axylia putris, andAutographa gamma, which were caught with some of the components and their blends.