The European Virus Archive: a new resource for virology research.

The European Virus Archive (EVA) was conceived as a direct response to the need for a coordinated and readily accessible collection of viruses that could be made available to academia, public health organisations and industry, initially within Europe, but ultimately throughout the world. Although scientists worldwide have accumulated virus collections since the early twentieth century, the quality of the collections and the viruses collected may vary according to the personal interests and agenda of the scientists. Moreover, when laboratories are re-organised or closed, collections are no longer maintained and gradually cease to exist. The tragedy of 9/11 and other disruptive activities have also meant that some previously available biological reagents are no longer openly exchanged between countries. In 2008, funding under the FP7-EU infrastructure programme enabled the initiation of the EVA. Within three years, it has developed from a consortium of nine European laboratories to encompass associated partners in Africa, Russia, China, Turkey, Germany and Italy. There is every reason to believe that EVA will continue to expand and ultimately exist as a globally networked, quality-controlled non-profit archive for the benefit of science. Organizations or individuals who would like to be considered as contributors are invited to contact the EVA coordinator, Jean-Louis Romette, at jean-louis.romette@univmed.fr.

Normative advice to campus crime victims: effects of gender, age, and alcohol.

Three studies investigated the appropriateness of calling the police as a function of crime, victim, and subject factors. In particular, the studies focused on whether and how the victim's consumption of alcohol affected normative advice to report the crime, as opposed to other options. Across the three studies, subjects viewed reporting as more appropriate for female victims, for victims who were 21 or older, and for victims who had not been drinking. In addition, females were more likely than males to believe reporting to the police was appropriate whereas males were more likely than females to favor some type of private action. Subjects viewed reporting as particularly inappropriate when the victim was underage and had been drinking. Results suggest that, because of the perceived stigma attached to victims who have been drinking, even serious victimizations may go unreported.

Clustering of haemagglutinin gene sequences of measles viruses isolated in the Gambia.

Partial nucleotide sequences of the haemagglutinin (H) gene of 18 measles viruses isolated in the Gambia during 1994 and 1995 show a high degree of homology (> 98.5%) when compared with each other. These sequences form a cluster distinct from measles viruses isolated from other geographic regions and from the sequences of vaccine strains and two isolates from the Gambia from earlier years. Despite the low level of genetic heterogeneity observed, a common feature of the Gambian isolates is the presence of three nucleotide changes (at positions 7963, 8649 and 8653) not observed in isolates from other locations.

Sequence variation within an outbreak of measles virus in the Coventry area during spring/summer 1993.

Measles virus (MV) was isolated from throat swab samples collected during the spring/summer 1993 in the Coventry area. Viral RNA was reverse transcribed and cDNA prepared using oligo- T primer. Using MV-specific primers the area encoding the external region of the haemagglutinin glycoprotein was amplified using nested PCR and cycle sequenced. Comparisons were made with the Edmonston strain and current MMR vaccine strain. It was found that a high degree of homology existed between all strains examined, but that a majority of clinical samples shared a premature termination signal that potentially shortened the haemagglutinin protein by 35 amino acids. The single clinical sample that lacked this early termination signal appeared to be closely related to the MMR strain and may result form a vaccine-related illness. Truncation of the haemagglutinin protein may have allowed MV to escape the immune response induced by vaccination with the current MMR vaccine.

IgG neutralization of type A influenza viruses and the inhibition of the endosomal fusion stage of the infectious pathway in BHK cells.

We have used three strains of type A influenza virus (A/fowl plaque virus/Rostock/34: H7N1; A/PR/8/34: H1N1; A/X31: H3N2) labeled with the fluorescent probe octadecyl rhodamine B chloride (R18) to study how neutralizing monoclonal IgG, specific for the haemagglutinin, affects the interaction of virus with BHK-21 cells. R18 labels viral lipid and self-quenches in the virion; dilution of R18 by fusion of viral and cellular membranes or by dissolution in detergent activates fluorescence. IgG neutralization reduced attachment to BHK-21 cells to an extent which depended on the virus-antibody concentration, but in no case was the inhibition of virus attachment to cells sufficient to fully account for the amount of neutralization obtained. The majority (> 78%) of attached IgG-neutralized virus became resistant to release by neuraminidase which indicated it was internalized by the cell. The endosomal fusion ability of IgG-neutralized virus which became cell-associated was also inhibited, to an extent which depended on the virus-antibody permutation. Inhibition of endosomal fusion, which is responsible for primary uncoating, was corroborated by immunofluorescence studies which showed that the NP antigen of IgG-neutralized virus, unlike that of infectious virus, did not accumulate in the nucleus. The loss of infectivity attributable to the combined inhibition of attachment and inhibition of fusion was sufficient to account for the extent of neutralization caused by relatively low concentrations of IgG (1/e or 63% neutralization). However it could not, with two possible exceptions, account for the neutralization resulting from higher concentrations of IgG (sufficient to cause > 99.9% neutralization), and we conclude that at least one other mechanism is operating simultaneously, such as that described by Rigg et al., (1989, J. Gen. Virol. 70, 2097-2109).

Morphological studies of the neutralization of influenza virus by IgM.

Quantitative relationships between neutralization, aggregation and attachment to monolayers of chick embryo fibroblast (CEF) cells have been studied using a constant amount of influenza A/fowl plague virus/Rostock/34 (H7N1) and varying amounts of purified mouse polyclonal IgM directed against the haemagglutinin, the major viral neutralization antigen. There are two major types of interaction. (i) At low concentrations of IgM there is aggregation of virus, but no neutralization provided that the aggregates are dispersed by vortexing and dilution. Maximum aggregation occurs at less than seven molecules of IgM per virion and the IgM is probably bound in the 'staple' or 'crab' conformation at these concentrations. (ii) At higher concentrations there is neutralization and this coincides with inhibition of attachment of virus to CEF cells. Neutralization of 50% infectivity requires about 35 molecules of IgM per virion. The maximum neutralization observed was only 87%. Quantitative data and electron microscopy observations suggest that molecules of IgM at the higher concentrations adopt a planar stance approximately perpendicular to the viral surface. It appears that IgM neutralizes fowl plague virus in vitro primarily by interfering with its attachment to cells; the fraction of neutralized virus that does attach is known not be internalized.

Mechanisms of neutralization of influenza virus in tracheal epithelial and BHK cells vary according to IgG concentration.

The interaction of IgG-neutralized type A influenza virus with differentiated epithelial cells of mouse trachea, BHK cells, and chicken erythrocytes was studied using three mouse monoclonal antibodies (IgG2a) each directed against a different antigenic site on the hemagglutinin. At high HIU:HAU ratios virus was neutralized greater than 99%, monodisperse, and attached to tracheal epithelial and BHK cells in normal amounts. The majority (70-80%) of neutralized virus failed to attach to erythrocytes. At low HIU:HAU ratios the virus was aggregated by each of the antibodies, and attachment to tracheal epithelial and BHK cells was inhibited by up to 75%. Combined aggregation and inhibition of attachment could theoretically account for up to 96% loss of infectivity but this corresponded with the observed degree of neutralization with only one of the antibodies. With increasing antibody:virus ratios, aggregation and inhibition of attachment contributed ever diminishingly to the observed neutralization and eventually not at all. Both neutralized and infectious virus attached to neuraminidase-sensitive receptors. After attachment neutralized virus became increasingly resistant to removal by neuraminidase suggesting that it had been internalized by the cell.

Mechanisms of neutralization of influenza virus on mouse tracheal epithelial cells by mouse monoclonal polymeric IgA and polyclonal IgM directed against the viral haemagglutinin.

The mechanism of neutralization of a type A influenza virus by polyclonal IgM was similar for both tracheal epithelial and BHK cells. Maximum neutralization was only 90% and most (70%) of the virus failed to attach to inoculated cells. The remainder attached to N-acetylneuraminic acid receptors but was not internalized. IgM aggregated virus, but only at an IgM: virus ratio below the level required for neutralization. Failure to detect any loss of infectivity associated with aggregation suggested that aggregates were unstable. Monoclonal polymeric IgA neutralized virus more efficiently on BHK cells (99.9%) than the equivalent amounts of IgM (90%). Otherwise the mechanisms of IgA and IgM neutralization were similar, except that IgA-induced aggregation was coincident with loss of infectivity and may thus have contributed to it. However, IgA-neutralized virus attached to tracheal epithelial cells more efficiently than infectious virus, initially using a neuraminidase-sensitive receptor, but then becoming neuraminidase-resistant. Whether the latter IgA-virus complexes were internalized or attached to a neuraminidase-resistant receptor is not known. This use of differentiated murine cells with murine IgA gave neutralization data that differed qualitatively from those obtained with the same antibody and undifferentiated hamster cells.

Prehospital care by EMTs and EMT-Is in a rural setting: prolongation of scene times by ALS procedures.

Because the initiation of IV lines by emergency medical technicians-Intermediates (EMT-Is) appeared to delay the patient's transport to the hospital, we undertook a retrospective study of 370 patients to compare prehospital care rendered by EMTs (EMT-A equivalent) and EMT-Is in a rural setting. Our study was limited to acute medical conditions in which protocols called for IV lines (124 patients with chest pain, 122 with acute respiratory distress, 99 with seizures, and only 25 with cardiac arrest) (the cardiac arrest cases were too few for statistical significance). We found that the difference in scene times for EMTs and EMT-Is not attempting IV lines was 6.1 and 6.9 minutes, respectively. The average scene time of EMT-Is attempting an IV line was 19.6 minutes (P less than .001) compared with EMT times, or times for EMT-Is not attempting an IV line. One hundred twenty-eight of 370 patients received IV medication within ten minutes of arrival in the emergency department, and ten of these patients had their IV lines initiated successfully in the field. Thirty-nine percent of patients with ED IV lines received IV medication within ten minutes of arrival, while only 21% of patients with a field IV line received medication in this period (P less than .05). We conclude that initiating a field IV line in this specific patient population significantly increased scene time and did not improve the chances of these patients receiving IV medication within ten minutes of arrival in the emergency department.

Glycosylated plasma protein measurement by a semi-automated method.

A semi-automated method for determination of glycosylation of plasma proteins using the thiobarbituric acid method is described. Plasma samples (100 microL) and a plasma pool used as a secondary standard were incubated in 0.3 M oxalic acid at 100 degrees C for exactly 2 h. The hydroxymethyl furfural released and the total protein were measured concurrently on a Technicon AAII Autoanalyzer. Addition of 10 or 20 mmol glucose to plasma samples caused a minimal increase in the measured glycosylated protein. Within-batch and between-batch coefficients of variation were 3.1% and 4.9% respectively. The mean glycosylated protein levels for 52 normals and 48 maturity-onset diabetics were (+/- 1SD) 0.96 +/- 0.13 and 1.75 +/- 0.32 nmol HMF/mg protein/2 h incubation.

Glycosylated haemoglobin: comparison of five different methods, including measurement on capillary blood samples.

Glycosylated haemoglobin was measured in venous blood samples and in blood collected in 'Unistep' bottles by isoelectric focusing (IEF), as the reference method, and by electroendosmosis (EEO), the thiobarbituric acid method (TBA), ion-exchange chromatography (IEC) and affinity chromatography (AC). Isoelectric focusing, electroendosmosis and thiobarbituric acid gave similar results. Affinity chromatography gave lower results than isoelectric focusing for normal values but similar results for diabetics. Ion-exchange chromatography gave 24% lower results than isoelectric focusing across the range. Using Unistep collected blood samples and comparing multiple samples from the same patient, electroendosmosis gave the best results (coefficient of variation 4%) and thiobarbituric acid gave slightly less good precision that other methods. Re-use of affinity chromatography columns gave less good precision. Collection of blood samples into a Unistep bottle gave similar results to venous sample results. Storage of venous capillary blood samples in Unistep bottles over 1 week at 21 degrees C gave similar results to immediate assay. Electroendosmosis of blood samples in Unistep bottles gave stable results over 2 weeks. Home collection by a patient of a capillary blood sample into a Unistep bottle allows glycosylated haemoglobin results to be available when seen in the clinic.

Lead in bone III. Prediction of social correlates from skeletal lead content in four Colonial American populations (Catoctin Furnace, College Landing, Governor's Land, and Irene Mound).

Lead content was determined in the skeletal tissue of 82 individuals representing two black and two white Colonial American populations: Catoctin Furnace, College Landing, Governor's Land, and Irene Mound. Group and individual differences in bone lead concentrations were used to assess behavioral, social and occupational characteristics. Variations in skeletal lead content suggested that the white owners of the Catoctin iron furnace shared little of their food and beverage with their black, male, industrial slaves, but that some of these workers' women had access to the owners' food sources--probably via domestic duty assignments. A broad range of lead concentrations in bones of the free blacks at College Landing implies a wide range of economic success among these tradesmen. Bone lead content of the white populations at Governor's Land and Irene Mound helped confirm family relationships that had been assigned on an archaeological and osteological basis, and also suggested that the social and functional status of the white tenant farmers' white servants frequently differed little from that of black slaves. These findings suggest that, when applied in appropriate circumstances, lead studies of archaeological skeletal tissue may provide information supplemental to that derived from historical, archaeological, or other conventional sources.