bluuebunny/crossref_metadata_2025_split huggingface dataset:
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

ABSTRACT“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

ABSTRACT
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

ABSTRACT
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

ABSTRACT

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

ABSTRACT

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

ABSTRACT

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

ABSTRACT
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

ABSTRACT
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

ABSTRACT

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

ABSTRACT

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

ABSTRACT
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.