bluuebunny/crossref_metadata_2025 huggingface dataset:
Dominant Epitopes of the C6 Diagnostic Peptide ofBorrelia burgdorferiAre Largely Inaccessible to Antibody on the Parent VlsE Molecule
Authors: Embers Monica E. , Jacobs Mary B. , Johnson Barbara J. B. , Philipp Mario T.
Published: June 2007

Lyme borreliosis (LB) is a disease for which antibody-based detection assays are often required for diagnosis. The variable surface molecule VlsE and IR6, one of its invariable regions, are commonly targeted by the antibody response in infected individuals. A series of enzyme-linked immunosorbent assays was performed to comparatively examine the antibody responses of North American LB patients (n= 37) to VlsE and invariable segments of this molecule. Both immunoglobulin M (IgM) and IgG responses to full-length VlsE and to peptides reproducing invariable regions 2, 4, and 6, as well as the invariable domains at the amino and carboxyl termini of VlsE, were assessed. The proportions and specificities of reactivity to the invariable segments were tested by using cognate peptides as competitors for VlsE binding by patient serum antibodies. IR6 epitopes (by the C6 peptide) were found to dominate the response to invariable segments. IR6 (C6)-specific antibodies were detected in 78% of the serum specimens, whereas <40% of patients generated antibodies that bound the N- or C-terminal domain and <12% of patients responded to either IR2 or IR4. Interestingly, 15 of 37 patients generated IgG antibodies that reacted with C6 but not with VlsE. Conversely, IgM responses were frequent for VlsE but not for invariable segments. A representative number of the serum specimens (n= 8) that contained IgG antibodies reacting with both C6 and VlsE was assessed in competition experiments, using C6 as a competitor. Only half of these specimens contained IgG antibodies whose binding to VlsE could be inhibited >50% by competition with the added C6 peptide. The median percent inhibition was 45.5%. These findings indicate that IR6 epitopes are largely concealed from the VlsE molecular surface and that full-length VlsE-based diagnosis likely detects antibodies to conformational and/or variable region epitopes.
The Bacterial Second Messenger cdiGMP Exhibits Promising Activity as a Mucosal Adjuvant
Authors: Ebensen Thomas , Schulze Kai , Riese Peggy , Morr Michael , Guzmán Carlos A.
Published: June 2007

The development of mucosal adjuvants is still a critical need in vaccinology. In the present work, we show that bis(3′,5′)-cyclic dimeric GMP (cdiGMP), a second messenger that modulates cell surface properties of several microorganisms, exerts potent activity as a mucosal adjuvant. BALB/c mice were immunized intranasally with the model antigen β-galactosidase (β-Gal) coadministered with cdiGMP. Animals receiving cdiGMP as an adjuvant showed significantly higher anti-β-Gal immunoglobulin G (IgG) titers in sera than controls (i.e., 512-fold [
P
< 0.05]). Coadministration of cdiGMP also stimulated efficient β-Gal-specific secretory IgA production in the lung (
P
< 0.016) and vagina (
P
< 0.036). Cellular immune responses were observed in response to both the β-Gal protein and a peptide encompassing its major histocompatibility complex class I-restricted epitope. The IgG1-to-IgG2a ratio of anti-β-Gal antibodies and the observed profiles of secreted cytokines suggest that a dominant Th1 response pattern is promoted by mucosal coadministration of cdiGMP. Finally, the use of cdiGMP as a mucosal adjuvant also led to the stimulation of in vivo cytotoxic T-lymphocyte responses in C57BL/6 mice intranasally immunized with ovalbumin and cdiGMP (up to 30% of specific lysis). The results obtained indicate that cdiGMP is a promising tool for the development of mucosal vaccines.
Clinical Evaluation of the SD Bioline Influenza Virus Antigen Test for Rapid Detection of Influenza Viruses A and B in Children and Adults during the Influenza Season
Authors: Yoo Young , Sohn Jang Wook , Park Dae Won , Kim Jeong Yeon , Shin Hye Kyung , Lee Yoon , Choung Ji Tae , Lee Chang Kyu , Kim Min Ja
Published: June 2007

The performance of the SD Bioline rapid antigen test kit for influenza virus detection was evaluated with 295 respiratory specimens during the influenza season. The overall sensitivity and specificity of the SD Bioline test were 61.9% and 96.8% for the influenza A virus antigen and 54.5% and 100% for the influenza B virus antigen, respectively. The results were consistent with peak influenza activities.
First Isolation of Cytopathogenic Bovine Torovirus in Cell Culture from a Calf with Diarrhea
Authors: Kuwabara Masaki , Wada Kazumasa , Maeda Yukiko , Miyazaki Ayako , Tsunemitsu Hiroshi
Published: June 2007

A cytopathogenic virus (designated the Aichi/2004 strain) was isolated in a human rectal adenocarcinoma cell line (HRT-18) from the ileum contents of a calf with diarrhea. Oval and elongated particles, approximately 100 to 170 nm in diameter, with club-shaped projections were seen in the infected culture supernatant, and torovirus-like (tubular and torus nucleocapsid) structures were seen in the infected cells by electron microscopy. An antiserum against bovine torovirus (BToV) reacted with the infected cells by immunofluorescence and neutralized the isolate. However, antisera against bovine coronavirus (BCV) failed to react with the infected cells by immunofluorescence or did not neutralize the isolate. Further, the isolate was positive for BToV by reverse transcription-PCR (RT-PCR) targeting fragments of the nucleocapsid (N), membrane (M), and spike (S) genes. Comparison of the nucleotide sequences of the PCR products with those of the published N, M, and S genes (476 to 497, 672, and 687 to 690 nucleotides, respectively) of toroviruses showed high sequence identities (up to 99.4%, 98.7%, and 94.9% for the N, M, and S genes, respectively) between the isolate and BToVs. In contrast, the isolate was negative for BCV by RT-PCR. In a serological survey of serum samples from 355 calves at 33 farms, 92% of calves were positive for neutralizing antibodies to the isolate. These results indicate that the isolate in this study was BToV and that BToV infection might be common in cattle in Japan. To our knowledge, this is the first isolation of BToV in tissue culture.
Comparison of Performance Characteristics of Three Real-Time Reverse Transcription-PCR Test Systems for Detection and Quantification of Hepatitis C Virus
Authors: Sábato M. Fernanda , Shiffman Mitchell L. , Langley Michael R. , Wilkinson David S. , Ferreira-Gonzalez Andrea
Published: June 2007

We evaluated the performance characteristics of three real-time reverse transcription-PCR test systems for detection and quantification of hepatitis C virus (HCV) and performed a direct comparison of the systems on the same clinical specimens. Commercial HCV panels (genotype 1b) were used to evaluate linear range, sensitivity, and precision. The Roche COBAS TaqMan HCV test for research use only (RUO) with samples processed on the MagNA Pure LC instrument (Roche RUO-MPLC) and Abbott analyte-specific reagents (ASR) with QIAGEN sample processing (Abbott ASR-Q) showed a sensitivity of 1.0 log10IU/ml with a linear dynamic range of 1.0 to 7.0 log10IU/ml. The Roche ASR in combination with the High Pure system (Roche ASR-HP) showed a sensitivity of 1.4 log10IU/ml with a linear dynamic range of 2.0 to 7.0 log10IU/ml. All of the systems showed acceptable reproducibility, the Abbott ASR-Q being the most reproducible of the three systems. Seventy-six clinical specimens (50 with detectable levels of HCV RNA and various titers and genotypes) were tested, and results were compared to those of the COBAS Amplicor HCV Monitor v2.0. Good correlation was obtained for the Roche RUO-MPLC and Abbott ASR-Q (R2= 0.84 andR2= 0.93, respectively), with better agreement for the Abbott ASR-Q. However, correlation (R2= 0.79) and agreement were poor for Roche ASR-HP, with bias relative to concentration and genotype. Roche ASR-HP underestimated HCV RNA for genotypes 3 and 4 as much as 2.19 log10IU/ml. Our study demonstrates that Roche RUO-MPLC and Abbott ASR-Q provided acceptable results and agreed sufficiently with the COBAS Amplicor HCV Monitor v2.0.
Is the Volume of Blood Cultured Still a Significant Factor in the Diagnosis of Bloodstream Infections?
Authors: Bouza Emilio , Sousa Dolores , Rodríguez-Créixems Marta , Lechuz Juan García , Muñoz Patricia
Published: June 2007

“The higher the volume of blood cultured the higher the yield of blood cultures” has been a well-accepted dictum since J. A. Washington II performed his classic work. This rule has not been questioned in the era of highly automated blood culture machines, nor has it been correlated with clinical variables. Our objective in this study was to complete a prospective analysis of the relationship between blood volume, the yield of blood cultures, and the severity of clinical conditions in adult patients with suspected bloodstream infections (BSI). During a 6-month period, random samples of blood cultures were weighed to determine the volume of injected blood (weight/density). Overall, 298 patients with significant BSI and 303 patients with sepsis and negative blood cultures were studied. The mean volume of blood cultured in patients with BSI (30.03 ± 14.96 ml [mean ± standard deviation]) was lower than in patients without BSI (32.98 ± 15.22 ml [P= 0.017]), and more episodes of bacteremia were detected with <20 ml (58.9%) than with >40 ml (40.2%) of blood cultured (P= 0.022). When patients were stratified according to the severity of their underlying condition, patients with BSI had higher APACHE II scores, and higher APACHE II scores were related to lower sample volumes (P< 0.001). A multivariate analysis showed that in the group of patients with APACHE II scores of ≥18, higher volumes yielded higher rates of bacteremia (odds ratio, 1.04 per ml of blood; 95% confidence interval, 1.001 to 1.08). We conclude that the higher yield of blood cultures inoculated with lower volumes of blood reflects the conditions of the population cultured. Washington's dictum holds true today in the era of automated blood culture machines.
Increased Detection Rate in Diagnosis of Herpes Simplex Virus Type 2 Meningitis by Real-Time PCR Using Cerebrospinal Fluid Samples
Authors: Franzen-Röhl Elisabeth , Tiveljung-Lindell Annika , Grillner Lena , Aurelius Elisabeth
Published: June 2007

Efficient and sensitive diagnostic methods are needed in the management of virus infections in the central nervous system. There is a demand for an evaluation of the sensitivity of PCR methods for early diagnosis of meningitis due to herpes simplex type 2 (HSV-2) and varicella-zoster virus (VZV). The objective of this study was to evaluate real-time PCR in the detection of HSV-2 and VZV DNA from cerebrospinal fluid (CSF) for etiological diagnoses in clinically well-characterized cases of primary and recurrent aseptic meningitis. Samples from 110 patients, 65 of whom were diagnosed with or were strongly suspected of having HSV-2 meningitis and 45 with acute aseptic meningitis of unknown causes, were analyzed. Results were compared with the outcome of nested PCR for HSV-2 infection. Clinical parameters were analyzed in relation to CSF viral load. With real-time PCR, HSV-2 DNA was found in CSF from 80% (52/65) of patients with clinical HSV-2 meningitis compared to 72% (47/65) found by nested PCR. The sensitivity of real-time HSV-2 PCR was found to be 87% (33/38) in primary and 70% (19/27) in recurrent meningitis. The HSV-2 viral load was significantly higher in primary than in recurrent meningitis and correlated with the degree of inflammation. VZV DNA was detected in 2 of 45 samples (4.4%). Real-time PCR for the diagnosis of HSV-2 meningitis was evaluated in a large, clinically well-characterized sample of material and found to identify more cases than nested PCR in the group of patients with recurrent meningitis. Quantification of DNA enables further research of disease prognosis and treatment.
Development and Validation of DNA Microarray for Genotyping Group A Rotavirus VP4 (P[4], P[6], P[8], P[9], and P[14]) and VP7 (G1 to G6, G8 to G10, and G12) Genes
Authors: Honma Shinjiro , Chizhikov Vladimir , Santos Norma , Tatsumi Masatoshi , do Carmo S. T. Timenetsky Maria , Linhares Alexandre C. , Mascarenhas Joana D'Arc P. , Ushijima Hiroshi , Armah George E. , Gentsch Jon R. , Hoshino Yasutaka
Published: June 2007

Previously, we reported the development of a microarray-based method for the identification of five clinically relevant G genotypes (G1 to G4 and G9) (V. Chizhikov et al., J. Clin. Microbiol. 40:2398-2407, 2002). The expanded version of the rotavirus microarray assay presented herein is capable of identifying (i) five clinically relevant human rotavirus VP4 genotypes (P[4], P[6], P[8], P[9], and P[14]) and (ii) five additional human rotavirus VP7 genotypes (G5, G6, G8, G10, and G12) on one chip. Initially, a total of 80 cell culture-adapted human and animal reference rotavirus strains of known P (P[1] to P[12], P[14], P[16], and P[20]) and G (G1-6, G8 to G12, and G14) genotypes isolated in various parts of the world were employed to evaluate the new microarray assay. All rotavirus strains bearing P[4], P[6], P[8], P[9], or P[14] and/or G1 to G6, G8 to G10, or G12 specificity were identified correctly. In addition, cross-reactivity to viruses of genotype G11, G13, or G14 or P[1] to P[3], P[5], P[7], P[10] to P[12], P[16], or P[20] was not observed. Next, we analyzed a total of 128 rotavirus-positive human stool samples collected in three countries (Brazil, Ghana, and the United States) by this assay and validated its usefulness. The results of this study showed that the assay was sensitive and specific and capable of unambiguously discriminating mixed rotavirus infections from nonspecific cross-reactivity; the inability to discriminate mixed infections from nonspecific cross-reactivity is one of the inherent shortcomings of traditional multiplex reverse transcription-PCR genotyping. Moreover, because the hybridization patterns exhibited by rotavirus strains of different genotypes can vary, this method may be ideal for analyzing the genetic polymorphisms of the VP7 or VP4 genes of rotaviruses.
Contribution of a New Mutation in
parE
to Quinolone Resistance in Extended-Spectrum-β-Lactamase-Producing
Escherichia coli
Isolates
Authors: Sorlozano Antonio , Gutierrez Jose , Jimenez Antonio , de Dios Luna Juan , Martínez José Luis
Published: June 2007

Mutations in the quinolone resistance-determining regions of
gyrA
,
gyrB
,
parC
, and
parE
were studied in 30 fluoroquinolone-resistant clinical isolates of
Escherichia coli
producing extended-spectrum β-lactamases. Ten isolates showed a mutation in
parE
that was significantly associated with an increase in the MIC for fluoroquinolones.
Detection of Four
Plasmodium
Species by Genus- and Species-Specific Loop-Mediated Isothermal Amplification for Clinical Diagnosis
Authors: Han Eun-Taek , Watanabe Risa , Sattabongkot Jetsumon , Khuntirat Benjawan , Sirichaisinthop Jeeraphat , Iriko Hideyuki , Jin Ling , Takeo Satoru , Tsuboi Takafumi
Published: June 2007

Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method, was developed for the clinical detection of four species of human malaria parasites:
Plasmodium falciparum
,
P. vivax
,
P. malariae
, and
P. ovale
. We evaluated the sensitivity and specificity of LAMP in comparison with the results of microscopic examination and nested PCR. LAMP showed a detection limit (analytical sensitivity) of 10 copies of the target 18S rRNA genes for
P. malariae
and
P. ovale
and 100 copies for the genus
Plasmodium
,
P. falciparum
, and
P. vivax
. LAMP detected malaria parasites in 67 of 68 microscopically positive blood samples (sensitivity, 98.5%) and 3 of 53 microscopically negative samples (specificity, 94.3%), in good agreement with the results of nested PCR. The LAMP reactions yielded results within about 26 min, on average, for detection of the genus
Plasmodium
, 32 min for
P. falciparum
, 31 min for
P. vivax
, 35 min for
P. malariae
, and 36 min for
P. ovale
. Accordingly, in comparison to the results obtained by microscopy, LAMP had a similar sensitivity and a greater specificity and LAMP yielded results similar to those of nested PCR in a shorter turnaround time. Because it can be performed with a simple technology, i.e., with heat-treated blood as the template, reaction in a water bath, and inspection of the results by the naked eye because of the use of a fluorescent dye, LAMP may provide a simple and reliable test for routine screening for malaria parasites in both clinical laboratories and malaria clinics in areas where malaria is endemic.
Sporadic “Transitional” Community-Associated Methicillin-Resistant
Staphylococcus aureus
Strains from Health Care Facilities in the United States
Authors: Brady Jennifer M. , Stemper Mary E. , Weigel Ashley , Chyou Po-Huang , Reed Kurt D. , Shukla Sanjay K.
Published: June 2007

We describe phenotypic and genotypic traits of a group of methicillin-resistant
Staphylococcus aureus
(MRSA) clones that are either remnants of unsuccessful community-associated MRSA (CA-MRSA) clones or represent a transitional state with some yet-to-be-acquired characteristics of CA-MRSA. These rare strains (
n
= 20) were identified during a 10-year period (1990-1999) from 13 unrelated health care facilities in Wisconsin. The isolates were recovered from patients in nosocomial or long-term chronic care facilities (60%) and outpatient settings (40%). Sixty percent (
n
= 12) of the isolates were recovered from skin and soft tissue infections, whereas the remaining isolates (
n
= 8) were from invasive infections. Ninety percent of isolates were susceptible to all antibiotic classes tested or resistant to erythromycin and clindamycin. Pulsed-field gel electrophoresis, multilocus sequence typing, and
spa
typing clustered these isolates into 8, 8, and 14 clonal groups, respectively. Eight plasmid profiles were represented in these strains. All four
agr
types were represented, with type IV being predominant (40%). All strains harbored subtypes of type IV staphylococcal cassette chromosome
mec
but lacked genes for the virulence factor Panton-Valentine leukocidin (PVL). The strains harbored one or more of the following toxin genes:
sea
,
seb
,
sec
,
sed
,
see
,
seh
,
sej
,
sek
,
sel
,
seg
,
sei
,
sem
,
sen
, and
seo
. Individual clonal groups maintained the same set of enterotoxin genes even though they were isolated over extended time periods, suggesting significant genomic stability. The potential role of PVL-carrying phages and plasmids in the success of CA-MRSA clones has been discussed.
Identification of a Novel Virulence Factor in Recombinant Pneumonia Virus of Mice
Authors: Krempl Christine D. , Wnekowicz Anna , Lamirande Elaine W. , Nayebagha Giw , Collins Peter L. , Buchholz Ursula J.
Published: June 2007

Pneumonia virus of mice (PVM) is a murine relative of human respiratory syncytial virus (HRSV). Here we developed a reverse genetics system for PVM based on a consensus sequence for virulent strain 15. Recombinant PVM and a version engineered to express green fluorescent protein replicated as efficiently as the biological parent in vitro but were 4- and 12.5-fold attenuated in vivo, respectively. The G proteins of HRSV and PVM have been suggested to contribute to viral pathogenesis, but this had not been possible to study in a defined manner in a fully permissive host. As a first step, we evaluated recombinant mutants bearing a deletion of the entire G gene (ΔG) or expressing a G protein lacking its cytoplasmic tail (Gt). Both G mutants replicated as efficiently in vitro as their recombinant parent, but both were nonpathogenic in mice at doses that would otherwise be lethal. We could not detect replication of the ΔG mutant in mice, indicating that its attenuation is based on a severe reduction in the virus load. In contrast, the Gt mutant appeared to replicate as efficiently in mice as its recombinant parent. Thus, the reduction in virulence associated with the Gt mutant could not be accounted for by a reduction in viral replication. These results identified the cytoplasmic tail of G as a virulence factor whose effect is not mediated solely by the viral load. In addition to its intrinsic interest, a recombinant virus that replicates with wild-type-like efficiency but does not cause disease defines optimal properties for vaccine development.
Critical Role of the 36-Nucleotide Insertion in Hepatitis B Virus Genotype G in Core Protein Expression, Genome Replication, and Virion Secretion
Authors: Li Ke , Zoulim Fabien , Pichoud Christian , Kwei Karen , Villet Stéphanie , Wands Jack , Li Jisu , Tong Shuping
Published: June 2007

Frequent coinfection of hepatitis B virus genotype G with genotype A suggests that genotype G may require genotype A for replication or transmission. In this regard, genotype G is unique in having a 12-amino-acid extension in the core protein due to a 36-nucleotide insertion near the core gene translation initiation codon. The insertion alters base pairing in the lower stem of the pregenome encapsidation signal, which harbors the core gene initiator, and thus has the potential to affect both core protein translation and pregenomic RNA encapsidation. Genotype G is also unusual for possessing two nonsense mutations in the precore region, which together with the core gene encode a secreted nonstructural protein called hepatitis B e antigen (HBeAg). We found that genotype G clones were indeed incapable of HBeAg expression but were competent in RNA transcription, genome replication, and virion secretion. Interestingly, the 36-nucleotide insertion markedly increased the level of core protein, which was achieved at the level of protein translation but did not involve alteration in the mRNA level. Consequently, the variant core protein was readily detectable in patient blood. The 12-amino-acid insertion also enhanced the genome maturity of secreted virus particles, possibly through less efficient envelopment of core particles. Cotransfection of genotypes G and A did not lead to mutual interference of genome replication or virion secretion. Considering that HBeAg is an immunotolerogen required for the establishment of persistent infection, its lack of expression rather than a replication defect could be the primary determinant for the rare occurrence of genotype G monoinfection.
The 24-Angstrom Structure of Respiratory Syncytial Virus Nucleocapsid Protein-RNA Decameric Rings
Authors: MacLellan Kirsty , Loney Colin , Yeo R. Paul , Bhella David
Published: June 2007

Respiratory syncytial virus (RSV), a nonsegmented, negative-sense RNA-containing virus, is a common cause of lower respiratory tract disease. Expression of RSV nucleocapsid protein (N) in insect cells using the baculovirus expression system leads to the formation of N-RNA complexes that are morphologically indistinguishable from viral nucleocapsids. When imaged in an electron microscope, three distinct types of structures were observed: tightly wound short-pitch helices, highly extended helices, and rings. Negative stain images of N-RNA rings were used to calculate a three-dimensional reconstruction at 24 Å resolution, revealing features similar to those observed in nucleocapsids from other viruses of the order
Mononegavirales
. The reconstructed N-RNA rings comprise 10 N monomers and have an external radius of 83 Å and an internal radius of 40 Å. Comparison of this structure with crystallographic data from rabies virus and vesicular stomatitis virus N-RNA rings reveals striking morphological similarities.
Porcine Arterivirus Attachment to the Macrophage-Specific Receptor Sialoadhesin Is Dependent on the Sialic Acid-Binding Activity of the N-Terminal Immunoglobulin Domain of Sialoadhesin
Authors: Delputte Peter L. , Van Breedam Wander , Delrue Iris , Oetke Cornelia , Crocker Paul R. , Nauwynck Hans J.
Published: June 2007

The sialic acid-binding lectin sialoadhesin (Sn) is a macrophage-restricted receptor for porcine reproductive and respiratory syndrome virus (PRRSV). To investigate the importance of pSn sialic acid-binding activity for PRRSV infection, an R
116
-to-E mutation was introduced in the predicted sialic acid-binding domain of pSn, resulting in a mutant, pSn
RE
, that could not bind sialic acids. PSn, but not pSn
RE
, allowed PRRSV binding and internalization. These data show that the sialic acid-binding activity of pSn is essential for PRRSV attachment to pSn and thus identifies the variable, N-terminal domain of Sn as a PRRSV binding domain.
In Vivo and In Vitro Escape from Neutralizing Antibodies 2G12, 2F5, and 4E10
Authors: Manrique Amapola , Rusert Peter , Joos Beda , Fischer Marek , Kuster Herbert , Leemann Christine , Niederöst Barbara , Weber Rainer , Stiegler Gabriela , Katinger Hermann , Günthard Huldrych F. , Trkola Alexandra
Published: June 2007

Recently, passive immunization of human immunodeficiency virus (HIV)-infected individuals with monoclonal antibodies (MAbs) 2G12, 2F5, and 4E10 provided evidence of the in vivo activity of 2G12 but raised concerns about the function of the two membrane-proximal external region (MPER)-specific MAbs (A. Trkola, H. Kuster, P. Rusert, B. Joos, M. Fischer, C. Leemann, A. Manrique, M. Huber, M. Rehr, A. Oxenius, R. Weber, G. Stiegler, B. Vcelar, H. Katinger, L. Aceto, and H. F. Gunthard, Nat. Med. 11:615-622, 2005). In the light of MPER-targeting vaccines under development, we performed an in-depth analysis of the emergence of mutations conferring resistance to these three MAbs to further elucidate their activity. Clonal analysis of the MPER of plasma virus samples derived during antibody treatment confirmed that no changes in this region had occurred in vivo. Sequence analysis of the 2G12 epitope relevant N-glycosylation sites of viruses derived from 13 patients during the trial supported the phenotypic evaluation, demonstrating that mutations in these sites are associated with resistance. In vitro selection experiments with isolates of four of these individuals corroborated the in vivo finding that virus strains rapidly escape 2G12 pressure. Notably, in vitro resistance mutations differed, in most cases, from those found in vivo. Importantly, in vitro selection with 2F5 and 4E10 demonstrated that resistance to these MAbs can be difficult to achieve and can lead to selection of variants with impaired infectivity. This remarkable vulnerability of the virus to interference within the MPER calls for a further evaluation of the safety and efficacy of MPER-targeting therapeutic and vaccination strategies.
Key factors for civilian injuries and deaths from exploding landmines and ordnance
Authors: Surrency Amber B , Graitcer Philip L , Henderson Alden K
Published: June 2007

Objective:To identify risk factors for death or injury from landmines and ordnance in Kabul City, Afghanistan, so programs can target preventive actions.

Methods:Active surveillance in hospitals and communities for injuries and deaths from landmine and ordnance explosions in Kabul City.

Results:Of the 571 people the authors identified during the 25-month period, 161 suffered a traumatic amputation and 94 were killed from a landmine or ordnance explosion. Of those asked, 19% of victims had received mine awareness education before the incident, and of those, the majority was injured while handling or playing with an explosive device. Most victims were young males with a few years of education. The occupation types most at risk were students and laborers, and unemployment was common among the victims. Collecting wood or paper and playing with or handling an explosive were the most frequent activities associated with injuries and deaths.

Conclusions:From May 1996 to July 1998, explosions from landmines and ordnance claimed 571 victims and were an important preventable cause of injury and death among people in Kabul City. Prevention strategies should focus on high-risk groups and changing risky behaviors, such as tampering with explosive devices.
Factors affecting the likelihood of reporting road crashes resulting in medical treatment to the police
Authors: Loo B P Y , Tsui K L
Published: June 2007

Objective:This paper aims to determine the percentage of road crashes resulting in injuries requiring hospital care that are reported to the police and to identify factors associated with reporting such crashes to the police.

Design:The data of one of two hospitals in the Road Casualty Information System were matched with the police’s Traffic Accident Database System. Factors affecting the police-reporting rate were examined at two levels: the different reporting rates among subgroups examined and tested with χ2tests; and multiple explanatory factors were scrutinised with a logistic regression model to arrive at the odds ratios to reflect the probability of police-reporting among subgroups.

Results:The police-reporting rate was estimated to be 57.5–59.9%. In particular, under-reporting among children (reporting rate = 33.6%) and cyclists (reporting rate = 33.0%) was notable.

Discussion:Accurate and reliable road crash data are essential for unveiling the full-scale and nature of the road safety problem. The police crash database needs to be supplemented by other data. In particular, any estimation about the social costs of road crashes must recognise the under-reporting problem. The large number of injuries not reflected in the police crash database represents a major public health issue that should be carefully examined.
Raised speed limits, case fatality and road deaths: a six year follow-up using ARIMA models
Authors: Friedman Lee S , Barach Paul , Richter Elihu D
Published: June 2007

Background:In November 1993, the Israeli government increased the speed limit for all vehicles from 90 to 100 km per hour on a total of 115 km of its three major interurban highways.

Design/Setting:We use ARIMA time series intervention models to evaluate the effect of the raise in speed limit on fatalities, serious injuries, and case-fatality for years 1988–1999. Motor vehicle crash data came from the Central Bureau of Statistics of Israel.

Results:Between January 1988 and December 1999, a total of 6029 persons were killed and 45 616 were seriously injured on roads in Israel. For all roads combined, the time series ARIMA model indicated that there were 4.69 more deaths per month (p<0.001), or 347 more than expected in the post-intervention period. Case-fatality rate (CFR) on all roads combined rose significantly (p<0.001). Modified case-fatality rate (CFRS) showed an increase of 2.5 deaths per 100 serious casualties (p<0.001).

Conclusions:The impact of raised speed limits was immediate and sustained. The largest increase in deaths occurred on interurban roads but a spillover effect was observed on urban roads as well. The increases in deaths and case-fatality rates persisted six years after the speed limit change despite major countermeasures and increasing congestion throughout the period of follow-up.
Testing the risk compensation hypothesis for safety helmets in alpine skiing and snowboarding
Authors: Scott Michael D , Buller David B , Andersen Peter A , Walkosz Barbara J , Voeks Jennifer H , Dignan Mark B , Cutter Gary R
Published: June 2007

Objective:The prevalence of helmet use by alpine skiers and snowboarders was estimated and self-reports on risk taking were assessed to test for potential risk compensation when using helmets in these sports.

Setting:Skiers and snowboarders were observed and interviewed at 34 resorts in the western United States and Canada.

Subjects:Respondents were 1779 adult skiers and snowboarders in the 2003 ski season.

Outcome measures:Observations of helmet use and questions about perceived speed and degree of challenge when not wearing a helmet (helmet wearers) or in previous ski seasons (non-helmet wearers).

Results:Helmet wearers reported that they skied/snowboarded at slower speeds (OR = 0.64, p<0.05) and challenged themselves less (OR = 0.76, p<0.05) than non-helmet wearers. Adoption of safety helmets in 2003 (23%) continued to increase over 2002 (OR = 0.46, p<0.05) and 2001 (OR = 0.84, p<0.05).

Conclusions:No evidence of risk compensation among helmet wearers was found. Decisions to wear helmets may be part of a risk reduction orientation. Helmet use continues to trend upwards but adoption may be slowing.
Quantifying the effect of a community-based injury prevention program in Queensland using a generalized estimating equation approach
Authors: Yorkston Emily , Turner Catherine , Schluter Philip J , McClure Rod
Published: June 2007

Objective:To develop a generalized estimating equation (GEE) model of childhood injury rates to quantify the effectiveness of a community-based injury prevention program implemented in 2 communities in Australia, in order to contribute to the discussion of community-based injury prevention program evaluation.

Design:An ecological study was conducted comparing injury rates in two intervention communities in rural and remote Queensland, Australia, with those of 16 control regions. A model of childhood injury was built using hospitalization injury rate data from 1 July 1991 to 30 June 2005 and 16 social variables. The model was built using GEE analysis and was used to estimate parameters and to test the effectiveness of the intervention.

Results:When social variables were controlled for, the intervention was associated with a decrease of 0.09 injuries/10 000 children aged 0–4 years (95% CI −0.29 to 0.11) in logarithmically transformed injury rates; however, this decrease was not significant (p = 0.36).

Conclusions:The evaluation methods proposed in this study provide a way of determining the effectiveness of a community-based injury prevention program while considering the effect of baseline differences and secular changes in social variables.
Hospital admissions following presentations to emergency departments for a fracture in older people
Authors: Boufous Soufiane , Finch Caroline , Close Jacqueline , Day Lesley , Lord Stephen
Published: June 2007

The aim of this paper is to estimate the proportion of older people who are hospitalised following a presentation to an emergency department for hip, pelvic and wrist fractures. The findings indicate that hospitalisation data do not accurately reflect the incidence of low-trauma fractures, particularly wrist and pelvic fractures, in older people.
Gun shows across a multistate American gun market: observational evidence of the effects of regulatory policies
Authors: Wintemute Garen J
Published: June 2007

Objective:To describe gun shows and assess the impact of increased regulation on characteristics linked to their importance as sources of guns used in crime.

Design:Cross-sectional, observational.

Subjects:Data were collected at a structured sample of 28 gun shows in California, which regulates these events and prohibits undocumented private party gun sales; and in Arizona, Nevada, Texas and Florida—all leading sources of California’s crime guns—where these restrictions do not exist.

Main outcome measures:Size of shows, measured by numbers of gun vendors and people in attendance; number and nature of guns for sale by gun vendors; measures of private party gun sales and illegal surrogate (“straw”) gun purchases.

Results:Shows in comparison states were larger, but the number of attendees per gun vendor was higher in California. None of these differences was statistically significant. Armed attendees were more common in other states (median 5.7%, interquartile range (IQR) 3.9–10.0%) than in California (median 1.1%, IQR 0.5–2.2%), p = 0.0007. Thirty percent of gun vendors both in California and elsewhere were identifiable as licensed firearm retailers. There were few differences in the types or numbers of guns offered for sale; vendors elsewhere were more likely to sell assault weapons (34.9% and 13.3%, respectively; p = 0.001). Straw purchases were more common in the comparison states (rate ratio 6.6 (95% CI 0.9 to 49.1), p = 0.06).

Conclusions:California’s regulatory policies were associated with a decreased incidence of anonymous, undocumented gun sales and illegal straw purchases at gun shows. No significant adverse effects of these policies were observed.
Workplace policies and prevalence of knee osteoarthritis: the Johnston County Osteoarthritis Project
Authors: Chen Jiu-Chiuan , Linnan Laura , Callahan Leigh F , Yelin Edward H , Renner Jordan B , Jordan Joanne M
Published: June 2007

Objective:

Previous studies on work and knee osteoarthritis (KOA) have been primarily focused on physical demands; very little is known about work-related organisational policies and KOA risks and outcomes. We examined the associations between workplace policies and KOA in a community-based population in the USA.

Methods:

The associations between employment offering accommodations (switch to physically less demanding jobs; part-time work for people needing reduced time) and benefits policies (paid sick leave; disability payment) with KOA outcomes (knee symptoms; symptomatic KOA [sKOA]; asymptomatic radiographic KOA [rKOA]) were analysed in participants (n = 1639) aged <65 years old and with completed employment histories and knee radiographs at baseline examination of the Johnston County Osteoarthritis Project. Multiple logistic regression models were used to estimate the prevalence odds ratios (ORs) of KOA associated with each workplace policy, adjusting for sociodemographic features, lifestyle factors, knee injuries, body mass index and other workplace characteristics. We used propensity score models to evaluate the differential selection in employment offering favourable policies and adjust for this potential bias accordingly.

Results:

Individuals employed in workplaces offering better policies had significantly less knee symptoms. Lower sKOA prevalence was noted in workplaces offering job-switch accommodation (8% vs. 13%), paid sick leave (9% vs. 16%) and disability payment (8% vs. 16%) than their counterparts. In multivariable models, the difference in sKOA prevalence was statistically significant for paid sick leave (adjusted OR 0.58, 95% CI 0.37 to 0.91) and disability payment policies (adjusted OR 0.54, 95% CI 0.35 to 0.85). Even among those without overt knee-related symptoms, a similar pattern of negative association between workplace policies and rKOA was present and remained robust after propensity score adjustment.

Conclusion:

The negative associations between KOA and workplace policies raise concerns about possible employment discrimination or beneficial effects of workplace policies. Longitudinal studies are needed to clarify the dynamic complexities of KOA risks and outcomes in relation to workplace policies.
Organic solvent exposure and hearing loss in a cohort of aluminium workers
Authors: Rabinowitz P M , Galusha D , Slade M D , Dixon-Ernst C , O’Neill A , Fiellin M , Cullen M R
Published: June 2007

Objectives:

Organic solvent exposure has been shown to cause hearing loss in animals and humans. Less is known about the risk of hearing loss due to solvent exposures typically found in US industry. The authors performed a retrospective cohort study to examine the relationship between solvent exposure and hearing loss in US aluminium industry workers.

Methods:

A cohort of 1319 workers aged 35 years or less at inception was followed for 5 years. Linkage of employment, industrial hygiene and audiometric surveillance records allowed for estimation of noise and solvent exposures and hearing loss rates over the study period. Study subjects were classified as “solvent exposed” or not, on the basis of industrial hygiene records linked with individual job histories. High frequency hearing loss was modelled as both a continuous and a dichotomous outcome.

Results:

Typical solvent exposures involved mixtures of xylene, toluene and/or methyl ethyl ketone (MEK). Recorded solvent exposure levels varied widely both within and between jobs. In a multivariate logistic model, risk factors for high frequency hearing loss included age (OR = 1.06, p = 0.004), hunting or shooting (OR = 1.35, p = 0.049), noisy hobbies (OR = 1.74, p = 0.01), baseline hearing level (OR = 1.04, p<0.001) and solvent exposure (OR = 1.87, p = 0.004). A multivariate linear regression analysis similarly found significant associations between high frequency hearing loss and age (p<0.001), hunting or shooting (p<0.001), noisy hobbies (p = 0.03), solvent exposure (p<0.001) and baseline hearing (p = 0.03).

Conclusion:

These results suggest that occupational exposure to organic solvent mixtures is a risk factor for high frequency hearing loss, although the data do not allow conclusions about dose–response relationships. Industries with solvent-exposed workers should include such workers in hearing conservation programs.
New insight into solvent-related end-stage renal disease: occupations, products and types of solvents at risk
Authors: Jacob S , Héry M , Protois J-C , Rossert J , Stengel B
Published: June 2007

Objectives:

It has been shown that all-solvent exposure is associated with the progression of primary glomerulonephritis to end-stage renal disease (ESRD), but little is known about the type of solvents that are high risk. The aim of this study was to investigate the role of solvents by occupation, product and type.

Methods:

Using a retrospective cohort design, the authors studied 269 patients with non-end-stage and biopsy-proven primary glomerulonephritis diagnosed between 1994 and 2001 in Paris and its suburbs. Two industrial hygienists evaluated patients’ exposures from lifetime occupational histories collected by interview from 2002–4, and using a list of the 30 most common solvents. The studied outcome was ESRD, defined as glomerular filtration rate <15 ml/mn/1.73 m2or dialysis. It was recorded during a mean follow-up of five years. Cox models were used to estimate adjusted hazard ratios (HR) of ESRD related to exposures.

Results:

Eighteen per cent of the patients had ever been exposed to solvents. Those with the highest risk of progression to ESRD were exposed machinery fitters and machine assemblers (HR 4.7, 95% CI 1.2 to 17.4) and plumbers/welders (HR 4.2, 95% CI 1.3 to 13.6), as compared to never exposed patients, as well as those who ever handled printing inks and petroleum products (HR 12.6 (95% CI 1.7 to 94.9) and 3.2 (95% CI 1.4 to 7.2), respectively). Among solvents, the highest risks were found for: toluene/xylene (HR 5.1, 95% CI 1.8 to 14.8), gasoline, fuel and gas-oil (HR 8.6, 95% CI 2.7 to 27.4), and ketones (HR 13.3, 95% CI 1.4 to 123.5).

Conclusion:

This study highlights the potential nephrotoxicity of several solvents. Intervention to promote screening for proteinuria in exposed workers may prevent the progression of glomerulonephritis to ESRD.
Correlations of geographic distribution and temperature of embryonic development with the nuclear DNA content in the Salamandridae (Urodela, Amphibia)
Authors: Litvinchuk Spartak N. , Rosanov Jury M. , Borkin Leo J.
Published: June 2007

We used flow cytometry to measure the nuclear DNA content in erythrocytes of 27 salamandrid species. Across these species, diploid genome size varied more than 2 fold (51.3–104.4 pg). According to genome size and geographic distribution, 3 groups of newt species were recognized: West Palearctics with smaller amounts of nuclear DNA; Nearctic, with intermediate values; and East Asiatic, with higher genome sizes. Viviparous West Palearctic salamanders differed from most of the oviparous West Palearctic newts in possessing larger genome sizes. The nuclear DNA content strongly correlates with species range limits. At the same temperature, embryos of salamandrid species with larger genome sizes have a markedly longer developmental time than those with smaller genomes. We present an analysis of the relationships between the amount of nuclear DNA and water temperature at the breeding sites.
Screen for new mutations on the 2nd chromosome involved in indirect flight muscle development in Drosophila melanogaster
Authors: Babu Sajesh , Ramachandra Nallur B.
Published: June 2007

An extensive ethylmethanesulfonate mutagenesis of Drosophila melanogaster was undertaken to isolate the stronger alleles of 3 indirect flight-muscle mutations. We isolated 17 strong mutant lines, with nearly complete penetrance and expressivity, using direct screening under polarized light, from more than 1700 mutagenized chromosomes. On complementation, we found 11 of these 17 mutant lines to be alleles of 3 indirect flight-muscle mutations (Ifm(2)RU1, 3 noncomplementing lines; ifm(2)RU2, 6 alleles; ifm(2)RU3, 2 alleles) of the previously isolated 8 complementation groups (Ifm(2)RU1to ifm(2)RU8). In addition, we found 6 new complementation groups with strong defects in adult-muscle morphology; we named these ifm(2)RS1 to ifm(2)RS6. All mutant lines were mapped by meiotic recombination, and 5 of the 6 new complementation lines were mapped using chromosome deficiencies. ifm(2)RS1 maps to a region that harbors ifm(2)RU4 (a mutation that was isolated previously); however, theses are not alleles because each complements the other mutation, and the mutant-muscle phenotype is very different. We used direct screening under polarized light to find recessive mutations; although this method was labor intensive, it can be used to identify recessive genes involved in myogenesis, unlike screens for flightlessness or wing-position defects. This screen identifies regions on the second chromosome that harbor probable genes that are likely expressed in the mesoderm and are thought to be involved in myogenesis. This screen has generated valuable resources that will help us to understand the role of many molecular players involved in myogenesis.
Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.)
Authors: Febrer Melanie , Cheung Foo , Town Christopher D. , Cannon Steven B. , Young Nevin D. , Abberton Michael T. , Jenkins Glyn , Milbourne Dan
Published: June 2007

White clover ( Trifolium repens L.) is a forage legume widely used in combination with grass in pastures because of its ability to fix nitrogen. We have constructed a bacterial artificial chromosome (BAC) library of an advanced breeding line of white clover. The library contains 37 248 clones with an average insert size of approximately 85 kb, representing an approximate 3-fold coverage of the white clover genome based on an estimated genome size of 960 Mb. The BAC library was pooled and screened by polymerase chain reaction (PCR) amplification using both white clover microsatellites and PCR-based markers derived from Medicago truncatula , resulting in an average of 6 hits per marker; this supports the estimated 3-fold genome coverage in this allotetraploid species. PCR-based screening of 766 clones with a multiplex set of chloroplast primers showed that only 0.5% of BAC clones contained chloroplast-derived inserts. The library was further evaluated by sequencing both ends of 724 of the clover BACs. These were analysed with respect to their sequence content and their homology to the contents of a range of plant gene, expressed sequence tag, and repeat element databases. Forty-three microsatellites were discovered in the BAC-end sequences (BESs) and investigated as potential genetic markers in white clover. The BESs were also compared with the partially sequenced genome of the model legume M. truncatula with the specific intention of identifying putative comparative-tile BACs, which represent potential regions of microsynteny between the 2 species; 14 such BACs were discovered. The results suggest that a large-scale BAC-end sequencing strategy has the potential to anchor a significant proportion of the genome of white clover onto the gene-space sequence of M. truncatula.