[Increasing Number of Road Traffic Fatalities in Germany - Turnaround or Snap-Shot]. / Warum wieder mehr Menschen im Straßenverkehr sterben ­ Trendwende oder Ausreißer?

Introduction: For the first time since 20 years, the number of road accident fatalities in 2011 increased on German roads compared to earlier periods. Methods and Results: The presented paper submitted by the expert group for accident prevention investigates and discusses possible reasons for the observed increase in road traffic fatalities. Results: Climate changes as well as changes in economic environment, and technological progress in car and passenger safety are identified as possible reasons for the observed increase. Discussion: Mentioning the "Decade of Action for Road Safety" initiated by the UNO and coordinated by the WHO, the overall goal is a worldwide reduction of accident related road fatalities. But prognostic calculations predict an asymptotic approximation to a limit of road fatalities. To achieve a reduction by half until 2020 intense collaboration and disproportional expenditure are necessary. Conclusion: From the authors' point of view the current increase of traffic fatalities in Germany is rated as a snapshot rather than a turnaround.

Whiplash-Associated Disorders : Clinical and medico-legal guidelines on the methods of ascertainment.

The manuscript presents the International Guidelines developed by the Working Group on Personal Injury and Damage under the patronage of the International Academy of Legal Medicine (IALM) regarding the Methods of Ascertainment of any suspected Whiplash-Associated Disorders (WAD).The document includes a detailed description of the logical and methodological steps of the ascertainment process as well as a synoptic diagram in the form of Flow Chart.

Muscle pathologies after cervical spine distortion-like exposure--a porcine model.

OBJECTIVE: Histological evaluation of porcine posterior cervical muscles after a forceful translational and extensional head retraction simulating high-speed rear end impact. METHODS: Four anesthetized pigs were exposed to a cervical spine distortion (CSD)-like motion in a lying position. After 2 different survival times of 4 and 6 h (posttrauma), the pigs were euthanized and tissue sampling of posterior cervical muscles was performed. A standard histological staining method involving paraffin-embedded sections was used to analyze the muscles, focusing on injury signs like hemorrhage and inflammatory cell reaction. A pig that was not subjected to impact was used as a control pig and was subjected to the same procedure to exclude any potential artifacts from the autopsy. RESULTS: The differentiation of 8 different posterior neck muscles in the dissection process was successful in more than 50 percent for each muscle of interest. Staining and valid analysis was possible from all extracted samples. Muscle injuries to the deepest posterior neck muscles could be found, especially in the musculus obliquus samples, which showed laminar bleedings in 4 out of 4 samples. In addition, in 4 out of 4 samples we were able to see increased cellular reactions. The splenius muscle also showed bleeding in all 4 samples. All animals showed muscle injury signs in more than three quarters of analyzed neck muscles. Differences between survival times of 4 and 6 h in terms of muscular injury were not of primary interest and could not be found. CONCLUSIONS: By simulating a CSD-like motion we were able to confirm injuries in the posterior cervical muscles under severe loading conditions. Further studies need to be conducted to determine whether these muscle injuries also occur under lower exposure forces.

Series of exon-skipping events in the elastic spring region of titin as the structural basis for myofibrillar elastic diversity.

Titins are megadalton-sized filamentous polypeptides of vertebrate striated muscle. The I-band region of titin underlies the myofibrillar passive tension response to stretch. Here, we show how titins with highly diverse I-band structures and elastic properties are expressed from a single gene. The differentially expressed tandem-Ig, PEVK, and N2B spring elements of titin are coded by 158 exons, which are contained within a 106-kb genomic segment and are all subject to tissue-specific skipping events. In ventricular heart muscle, exons 101 kb apart are joined, leading to the exclusion of 155 exons and the expression of a 2.97-MDa cardiac titin N2B isoform. The atria of mammalian hearts also express larger titins by the exclusion of 90 to 100 exons (cardiac N2BA titin with 3.3 MDa). In the soleus and psoas skeletal muscles, different exon-skipping pathways produce titin transcripts that code for 3.7- and 3.35-MDa titin isoforms, respectively. Mechanical and structural studies indicate that the exon-skipping pathways modulate the fractional extensions of the tandem Ig and PEVK segments, thereby influencing myofibrillar elasticity. Within the mammalian heart, expression of different levels of N2B and N2BA titins likely contributes to the elastic diversity of atrial and ventricular myofibrils.

When and why do people avoid unknown probabilities in decisions under uncertainty? Testing some predictions from optimal foraging theory.

When given a choice between two otherwise equivalent options - one in which the probability information is stated and another in which it is missing - most people avoid the option with missing probability information (Camerer & Weber, 1992). This robust, frequently replicated tendency is known as the ambiguity effect. It is unclear, however, why the ambiguity effect occurs. Experiments 1 and 2, which separated effects of the comparison process from those related to missing probability information, demonstrate that the ambiguity effect is elicited by missing probabilities rather than by comparison of options. Experiments 3 and 4 test predictions drawn from the literature on behavioral ecology. It is suggested that choices between two options should reflect three parameters: (1) the need of the organism, (2) the mean expected outcome of each option; and (3) the variance associated with each option's outcome. It is hypothesized that unknown probabilities are avoided because they co-occur with high outcome variability. In Experiment 3 it was found that subjects systematically avoid options with high outcome variability regardless of whether probabilities are explicitly stated or not. In Experiment 4, we reversed the ambiguity effect: when participants' need was greater than the known option's expected mean outcome, subjects preferred the ambiguous (high variance) option. From these experiments we conclude that people do not generally avoid ambiguous options. Instead, they take into account expected outcome, outcome variability, and their need in order to arrive at a decision that is most likely to satisfy this need.

The ORF, regulated synthesis, and persistence-specific variation of influenza C viral NS1 protein.

The open reading frame (ORF) and the regulated synthesis of the influenza C viral NS1 protein were analyzed in view of viruses possessing different biological activities. We provide evidence for a 246-amino-acid NS1-ORF, encoded by five viral strains and variants. Prokaryotic expression of the prototype NS1-ORF resulted in a product of 27 kDa, confirming the predicted molecular weight. Using an antiserum raised against recombinant NS1 protein, nonstructural proteins of wild-type virus were detected in infected cells for a limited course of time, whereas a persistent virus variant was characterized by a long-term nonstructural gene expression. As examined by infection experiments, the intracellular distribution of nonstructural protein was nuclear and cytoplasmic, whereas in NS1 gene-transfected cells, the cytoplasmic localization occurred in a fine-grained structure, suggesting an analogy to influenza A viral NS1 protein. Concerning persistent infection, NS1 protein species differing in sizes and posttranslational modifications were observed for a persistent virus variant, as particularly illustrated by a high degree of NS1 phosphorylation. Virus reassortant analyses proved the importance of the NS-coding genomic segment: the minimal viral properties required for the establishment of persistence were transferred with this segment to a monoreassortant virus. Thus the influenza C viral NS1 protein is a 246-amino-acid nuclear-cytoplasmic phosphoprotein that can be subject to specific variations being functionally linked to a persistent virus phenotype.

Effects of subinhibitory concentrations of antibiotics on alpha-toxin (hla) gene expression of methicillin-sensitive and methicillin-resistant Staphylococcus aureus isolates.

Concentrations of antibiotics below the MIC are able to modulate the expression of virulence-associated genes. In this study, the influence of subinhibitory doses of 31 antibiotics on the expression of the gene encoding the staphylococcal alpha-toxin (hla), a major virulence factor of Staphylococcus aureus, was investigated with a novel gene fusion protocol. The most striking observation was a strong induction of hla expression by subinhibitory concentrations of beta-lactams and an almost complete inhibition of alpha-toxin expression by clindamycin. Whereas glycopeptide antibiotics had no effect, the macrolide erythromycin and several aminoglycosides reduced and fluoroquinolones slightly stimulated hla expression. Furthermore, Northern blot analysis of hla mRNA and Western blot (immunoblot) analysis of culture supernatants of both methicillin-sensitive and methicillin-resistant S. aureus strains revealed that methicillin-induced alpha-toxin expression is a common phenomenon of alpha-toxin-producing strains. Some methicillin-resistant S. aureus isolates produced up to 30-fold more alpha-toxin in the presence of 10 microg of methicillin per ml than in its absence. The results indicate that the novel gene fusion technique is a useful tool for studying the modulation of virulence gene expression by antibiotics. Moreover, the results suggest that the effects of certain antibiotics on virulence properties may be relevant for the management of S. aureus infections.

Cloning of aas, a gene encoding a Staphylococcus saprophyticus surface protein with adhesive and autolytic properties.

A gene encoding a novel cell wall-associated protein of Staphylococcus saprophyticus that binds fibronectin and to sheep erythrocytes has been cloned and sequenced. The 4392 bp open reading frame codes for an amino acid sequence that is quite similar to the Atl, an autolysin, of Staphylococcus aureus and to the AtlE of S. epidermidis. The two regions of most pronounced homology code for an N-acetyl-muramyl-L-alanine amidase and for an endo-beta-N-acetyl-D-glucosaminidase. The cloned protein lysed cells of S. saprophyticus and Micrococcus luteus exogenously. Subcloning localized the enzymatic activities to the regions of high homology and demonstrated that the interposed sequence is responsible for the adhesive activities. Two allelic replacement mutants were constructed that lacked autolytic activity and adhesive properties. The N-terminal portion of the protein contains seven highly conserved, contiguous repeats with no similarity to published sequences. It lacks the motifs typical of Gram-positive surface proteins and shows a different overall organization. This autolysin/adhesin of S. saprophyticus (Aas) appears to represent a new class of staphylococcal adhesins.

Atherogenic properties of enzymatically degraded LDL: selective induction of MCP-1 and cytotoxic effects on human macrophages.

The mechanisms underlying the selective accumulation of macrophages in early atherosclerotic lesions are poorly understood but are likely to be related to specific properties of altered low density lipoprotein (LDL) deposited in the subendothelium. Enzymatic, nonoxidative degradation of LDL converts the lipoprotein to a potentially atherogenic moiety, enzymatically altered LDL (E-LDL), which activates complement and is rapidly taken up by human macrophages via a scavenger receptor-dependent pathway. Immunohistological evidence indicates that E-LDL is present in an extracellular location in the early lesion. We report that E-LDL causes massive release of monocyte chemotactic protein 1 (MCP-1) from macrophages and that expression of interleukin 8 or RANTES remains unchanged. Release of MCP-1 was preceded by a rapid expression of MCP-1 mRNA, which was detectable after 15 minutes, reached maximum levels after 1 hour, and remained detectable for 12 hours after exposure to concentrations as low as 10 microg/mL E-LDL. MCP-1 mRNA induction and protein release by E-LDL exceeded that evoked by oxidized LDL. Release of MCP-1 was dependent on de novo protein synthesis and on the activity of tyrosine kinases. At higher concentrations, E-LDL, but not oxidized LDL, exerted toxic effects on macrophages that in part appeared to be due to apoptosis. The results show that E-LDL possesses major properties of an atherogenic lipoprotein.

Mutational analysis of the genes encoding urokinase-type plasminogen activator (uPA) and its inhibitor PAI-1 in advanced ovarian cancer.

Evidence has accumulated that urokinase-type plasminogen activator (uPA), its inhibitor (PAI-1) and receptor (uPAR) are involved in tumor invasion and metastasis. We analyzed the DNA sequences encoding these factors to see if they are altered in the ovarian cancer cell lines OV-MZ-6, OV-MZ-19, and OVCAR-3. In the uPA-encoding cDNA derived from OV-MZ-6 cells (but not in the uPA-cDNA from OVCAR-3 and OV-MZ-19), a so-far unknown mutation was identified in codon 121, resulting in a proline to leucine exchange. This exchange creates an AluI restriction site making restriction fragment length polymorphism (RFLP) analyses possible. Previously published PAI-1 sequences pointed to a variation of amino acid 15 of the PAI-1 signal sequence representing either threonine or alanine, which was confirmed in the present study. The uPAR cDNAs of all three cell lines encoded the published wild-type sequence. In order to elucidate the possible role of the Pro121Leu exchange in uPA and the Ala/Thr variants in the signal sequence of PAI-1 in the development and/or progression of human ovarian cancer, we studied the presence of these mutants or variants in a series of 22 ovarian cancer tissues. In addition to the wild-type sequence, the Pro121Leu exchange in the uPA sequence was detected in 10 out of 22 tumor tissues; 11 tumors carried exclusively the Pro121 allele; in one case exclusively the Leu121 allele was detected. In 18/22 tumors, triplet 15 in the signal sequence of PAI-1 encoded alanine, four DNAs contained both the Ala and the Thr allele. Furthermore, we analyzed another known common single-base-pair insertion/deletion polymorphism (ins/del allele) found in the promoter region of the PAI-1 gene and thought to be of functional importance in regulating PAI-1 gene expression. The PAI-1 ins-allele was found in 3/22, the del-allele in 6/22 and both alleles in 13/22 ovarian cancer tissues. In genomic DNA isolated from peripheral blood of 23 healthy donors, we observed similar allele frequencies of the three polymorphisms as found in the 22 ovarian carcinomas. Taken together, these results suggest that the polymorphisms observed in the uPA and PAI-1 genes may not be linked to ovarian cancer.

[Injuries to the cervical spine in automobile accidents]. / Verletzungen der Halswirbelsäule bei Pkw-Unfällen.

Despite the improvement of protective properties in vehicles acceleration injuries to the cervical spine have not lost their significance. The increase of minor injuries and a high rate of seat belt usage is, beside other reasons, responsible for the increase of mild cervical spine injuries. From a data pool of 15,000 vehicle-vehicle collisions the portion of cervical spine injuries was selected for further investigation. Acceleration injuries to the cervical spine occur especially during rear end collisions. It can be shown that women, front seat occupants and occupants of lighter vehicles have a higher risk of suffering from such injuries. More than 90% of cervical spine injured had mild injuries by AIS standards; the portion of injuries with fractures was 0.6%. The diagnosis "wiplash injury" or "acceleration injury to the cervical spine" was given in more than 30% of all cervical spine injured showing only 1 symptom. Although most patients are first being treated in a hospital only 2/3 from 744 detailed studied cases were x-rayed, which in 71% were without significant conditions. Additional instrumental diagnostic measures (CT, MR) were used in only 1%. The study indicates that the "acceleration injury" requires a critical verification by doctors and insurers. A detailed finding following a thorough history taking and, in case, an interdisciplinary definition of accident injuries an early point of time will certain the diagnosis and facilitate therapy and judgement.

A non-radioactive multiprime sequencing method for HIV genomes.

A manual non-radioactive DNA sequencing protocol was developed for rapid analysis of variable HIV-1 genomes. Sets of up to ten primers were used in one sequencing reaction. After polyacrylamide gel electrophoresis and blotting onto nylon membranes the individual sequences were detected by hybridization with digoxigenin-labelled oligonucleotides and chemiluminescence. The method is applicable to any sequencing project where numerous variants of DNA fragments of several 1000 bp of length are to be analysed.

[Falling asleep at the wheel: the chief cause of severe traffic accidents]. / Einschlafen am Steuer: Hauptursache schwerer Verkehrsunfälle.

All fatal traffic accidents on highways in Bavaria in 1991 have been analyzed. The most frequent single cause for the 204 accidents was falling asleep (24%). A time of day analysis revealed that the highest accidents frequencies could be observed at 6:00 a.m. and 2:00 p.m. In addition an age effect could be observed. The majority of the day accidents were caused by older drivers, while most of the night accidents were caused by younger ones.

[Generalized nocardiosis with meningoencephalitis in a nonimmunosuppressed female patient]. / Generalisierte Nokardiose mit Meningoenzephalitis bei einer nicht-immunsupprimierten Patientin.

Four weeks after an attack of pneumonia of unknown aetiology a 40-year-old woman was hospitalized because of a nonpurulent, predominantly basal meningoencephalitis and infratentorial abscesses. She had dysarthria, mild right-sided motor hemiparesis and central paresis affecting the 7th cranial nerve. An area of fluctuating resistance, about 3 cm in diameter, was noticed over the left thigh. Serology indicated inflammatory disease, but there was no immunodeficiency. The CSF showed lymphocytic pleocytosis with mild protein increase but no evidence of infective agent. As tubercular meningitis was suspected she was treated with rifampicin (300 mg i.v. twice daily), isoniazid (300 mg i.v. once daily), streptomycin (800 mg i.m. once daily), cefotaxime (2.0 g i.v. three times daily), fluconazole (200 mg i.v. once daily) and dexamethasone (16-8-8 mg i.v.). She suddenly died two days after admission, probably as the result of central regulatory failure. Generalized nocardiosis involving lung, subcutaneous tissue and brain was revealed at autopsy. Although nocardiosis occurs predominantly in patients under immunosuppression, this infection should be considered in the differential diagnosis of treatment-resistant pneumonia and meningoencephalitis without obvious predisposition.

[Legionellosis in a newborn infant]. / Legionellose bei einem Neugeborenen.

We describe a case of successful therapy of a neonatal Legionellosis with Erythromycin. On his 6th day of life a full term newborn with normal body weight was affected by a severe pneumonia. This was at first resistant to therapy and required mechanical ventilation. Diagnosis of Legionella pneumophila serotype 1 was made by culture from bronchial lavage. Only few cases of neonatal Legionellosis have been reported until now. In three cases diagnosis was made post mortem.

Cleavage of tumor necrosis factor-alpha by Legionella exoprotease.

The role of the major secretory protein of Legionella pneumophila, a zinc protease, in Legionella infection is not known. Since an important step of the host reaction in Legionnaires' disease is the production of tumor necrosis factor-alpha (TNF-alpha) by alveolar macrophages, we studied the interaction of Legionella protease and U-937 cells with respect to TNF-alpha. The Legionella protease was purified by fractionated precipitation, gel filtration and hydrophobic interaction chromatography. The purified enzyme was added to U-937 cells, a promyelocytic cell line. In the supernatants of PMA-treated U-937 cells we found low concentrations of TNF-alpha after incubation with protease. Therefore we pursued the hypothesis of direct enzymatic degradation of TNF-alpha by Legionella protease. Enzymatic cleavage of TNF-alpha was proven by SDS-PAGE, ELISA and TNF-alpha bioassay with L-929 cells. The degradation of TNF-alpha by the Legionella protease was shown in all three systems. Enzymatic degradation of TNF-alpha might be important for the pathogenesis of Legionnaires' disease.

[Morphologic HIV demonstration in formalin embedded material--techniques, problems, results]. / Morphologischer HIV-Nachweis an formalinfixiertem Material--Techniken, Probleme, Ergebnisse.

Formalin-fixed, paraffin-embedded material of archival specimens is suitable for a morphological HIV-detection: Infected cells with HIV in the proliferative phase can be demonstrated with reliable results on tissue sections by immunohistological technics using new antibodies. In situ nucleic acid hybridisation technics can also show HIV in the expression phase on paraffin-embedded material, but often fail in demonstrating latently HIV-infected cells. The DNA-Polymerase chain reaction can detect latent Provirus in morphologically defined areas of paraffin sections even in autopsy material, i.e. lymphnodes and even eyes of patients with HIV-Infection, but requires precaution and control with respect to contamination.

[A simple method for the demonstration of the bacterial spectrum in the nose and the nasopharynx in the infection-free interval in children with adenoid hypertrophy]. / Eine einfache Methode zum Nachweis des bakteriellen Keimspektrums in Nase und Nasenrachenraum im infektfreien Intervall bei Kindern mit Rachenmandelhyperplasie.

The implications of bacterial colonization and distribution patterns in the nasopharynx and nasal cavities of children with adenoidal hypertrophy without clinical signs of acute infection are to be determined. We examined the spectrum and distribution of the facultative pathogenic bacterial flora in nasal cavities and nasopharynx of children with clinical apparent symptoms or signs of adenoid hypertrophy in an infection free interval. Compared with the nasal cavity we found an accumulation of pathogenic bacteria in the nasopharynx. A transnasal single swab from the nasopharynx showed to be the most effectively practical way to detect clinically relevant pathogenic bacteria. A thin flexible calcium-alginate swab was used in our experiments. Swabbing from the anterior nasal cavities proved to be a minor successful diagnostic method.