bluuebunny/crossref_metadata_2025_split huggingface dataset:
Persistence of a Positive (1,3)-β-
d
-Glucan Test after Clearance of Candidemia in Hematopoietic Stem Cell Transplant Recipients
Authors: Mikulska M. , Furfaro E. , Del Bono V. , Gualandi F. , Van Lint M. T. , Miletich F. , Bacigalupo A. , Viscoli C. 
Published: January 2011

ABSTRACT

In 6 hematopoietic stem cell transplant (HSCT) recipients with candidemia, the (1,3)-β-
d
-glucan (BG) test was positive a median of 2.5 days after a positive blood culture. Only in 1 patient did BG positivity precede positive blood cultures. BG concentrations decreased in patients with clinical response, but positive BG results persisted long after blood cultures became sterile (median, 48 days).
ThertxAToxin Gene of Kingella kingae: a Pertinent Target for Molecular Diagnosis of Osteoarticular Infections
Authors: Lehours Philippe , Freydière Anne-Marie , Richer Olivier , Burucoa Christophe , Boisset Sandrine , Lanotte Philippe , Prère Marie Françoise , Ferroni Agnès , Lafuente Christine , Vandenesch Francois , Mégraud Francis , Ménard Armelle 
Published: January 2011

ABSTRACTKingella kingaeis an emerging osteoarticular pathogen in young children. Its isolation by traditional culture methods remains difficult, underscoring the need to implement other diagnostic methods for its detection and identification, such as nucleic acid amplification tests. Although the genome of this bacterium has not yet been sequenced, a toxin named RTX has been identified. The goal of this study was to develop sensitive, specific, and rapid molecular methods based on thertxAtoxin gene sequence to diagnose this infection. Two real-time PCR assays (SYBR green and TaqMan chemistries) targeting this gene are reported. Sensitivity and specificity were first evaluated successfully with 67 strains: 31Kingella kingaeisolates and 36 strains from other bacterial species. Then, 52 clinical specimens positive or negative by culture and/or PCR (16S rRNA andcpn60genes) were tested with these assays. A nested PCR assay with subsequent sequencing was also developed to confirm the presence ofKingella kingaeisolates in these clinical specimens. The results obtained demonstrate that these assays are accurate for the diagnosis ofKingella kingaeinfection.
Detection of Hepatitis C Virus and Antibodies in Postmortem Blood and Bloodstains
Authors: Takasaka Tomokazu , Itoh Yoshito , Kaneko Hiroto , Ikegaya Hiroshi 
Published: January 2011

ABSTRACT
To evaluate the risk of accidental hepatitis C virus (HCV) infection, we examined whether anti-HCV antibodies and HCV RNA were detectable in HCV-infected blood samples from living donors, cadavers, and bloodstains. We showed that even after blood has left the body for several days, anti-HCV antibodies and HCV RNA may persist in it.
Colorimetric-Plate Method for Rapid Disk Diffusion Susceptibility Testing of Escherichia coli
Authors: Sener Sehnaz , Acuner Ibrahim Cagatay , Bek Yuksel , Durupinar Belma 
Published: January 2011

ABSTRACT

Here, we report a laboratory-developed colorimetric-plate method for rapid disk diffusion susceptibility testing of
Escherichia coli
. One hundred isolates were evaluated. Categorical agreement between the colorimetric plate and the standard disk diffusion method was 99%. Mean time to results was 7.07 h (95% confidence interval, 5.96 to 8.19).
Outbreak of Infections by Hepatitis B Virus Genotype A and Transmission of Genetic Drug Resistance in Patients Coinfected with HIV-1 in Japan
Authors: Fujisaki Seiichiro , Yokomaku Yoshiyuki , Shiino Teiichiro , Koibuchi Tomohiko , Hattori Junko , Ibe Shiro , Iwatani Yasumasa , Iwamoto Aikichi , Shirasaka Takuma , Hamaguchi Motohiro , Sugiura Wataru 
Published: January 2011

ABSTRACTThe major routes of hepatitis B virus (HBV) infection in Japan has been mother-to-child transmission (MTCT) and blood transfusion. However, HBV cases transmitted through sexual contact are increasing, especially among HIV-1-seropositive patients. To understand the molecular epidemiology of HBV in HBV/HIV-1 coinfection, we analyzed HBV genotypes and HIV-1 subtypes in HBV/HIV-1-coinfected patients at Nagoya Medical Center from 2003 to 2007. Among 394 HIV-1-infected Japanese men having sex with men (MSM) who were newly diagnosed during the study period, 31 (7.9%) tested positive for the hepatitis B virus surface antigen. HBV sequence analyses were successful in 26 cases, with 21 (80.7%) and 5 (19.3%) cases determined as genotypes A and C, respectively. Our finding that HBV genotype A was dominant in HIV-1-seropositive patients alerts clinicians to an alternative outbreak of HBV genotype A in the HIV-1-infected MSM population and a shift in HBV genotype from C to A in Japan. The narrow genetic diversity in genotype A cases suggests that genotype A has been recently introduced into the MSM population and that sexual contacts among MSM were more active than speculated from HIV-1 tree analyses. In addition, we found a lamivudine resistance mutation in one naïve case, suggesting a risk of drug-resistant HBV transmission. As genotype A infection has a higher risk than infection with other genotypes for individuals to become HBV carriers, prevention programs are urgently needed for the target population.
Direct Genotyping of Toxoplasma gondii in Ocular Fluid Samples from 20 Patients with Ocular Toxoplasmosis: Predominance of Type II in France
Authors: Fekkar A. , Ajzenberg D. , Bodaghi B. , Touafek F. , Le Hoang P. , Delmas J. , Robert P. Y. , Dardé M. L. , Mazier D. , Paris L. 
Published: January 2011

ABSTRACT

We report the direct genotyping analysis of
Toxoplasma gondii
in ocular samples collected from 20 patients, as well as associated clinical and epidemiological data. This work was aimed at better understanding the impact of genotypes of
Toxoplasma gondii
strains on toxoplasmic retinochoroiditis. For this purpose, we studied the aqueous humor (AH) or vitreous humor (VH) of 20 patients presenting with ocular toxoplasmosis (OT) in 2 hospitals in France. Genetic characterization was obtained with microsatellite markers in a multiplex PCR assay. In contrast to the results of previous studies, we found no association between atypical
Toxoplasma gondii
genotypes and the occurrence of OT. Considering the local epidemiological data, our OT patients seemed to be infected more frequently by ordinary type II strains found in the environment. In conclusion, direct genotyping of
Toxoplasma gondii
strains from aqueous or vitreous humor showed a predominance of the type II genotype in ocular toxoplasmosis; this may be due to a high exposure rate of this genotype in humans.
Laboratory Diagnosis, Epidemiology, and Clinical Outcomes of Pandemic Influenza A and Community Respiratory Viral Infections in Southern Brazil
Authors: Raboni Sonia M. , Stella Vanessa , Cruz Cristina R. , França João B. , Moreira Suzana , Gonçalves Lili , Nogueira Meri B. , Vidal Luine R. , Almeida Sergio M. , Debur Maria C. , Carraro Hipolito , Duarte dos Santos Claudia N. 
Published: January 2011

ABSTRACT
Community respiratory viruses (CRVs) are commonly associated with seasonal infections. They have been associated with higher morbidity and mortality among children, elderly individuals, and immunosuppressed patients. In April 2009, the circulation of a new influenza A virus (FLUA H1N1v) was responsible for the first influenza pandemic of this century. We report the clinical and epidemiological profiles of inpatients infected with CRVs or with FLUA H1N1v at a tertiary care hospital in southern Brazil. In addition, we used these profiles to evaluate survivor and nonsurvivor patients infected with FLUA H1N1v. Multiplex reverse transcription-PCR (RT-PCR) and real time RT-PCR were used to detect viruses in inpatients with respiratory infections. Record data from all patients were reviewed. A total of 171 patients were examined over a period of 16 weeks. Of these, 39% were positive for FLUA H1N1v, 36% were positive for CRVs, and 25% were negative. For the FLUA H1N1v- and CRV-infected patients, epidemiological data regarding median age (30 and 1.5 years), myalgia (44% and 13%), need for mechanical ventilation (44% and 9%), and mortality (35% and 9%) were statistically different. In a multivariate analysis comparing survivor and nonsurvivor patients infected with influenza A virus H1N1, median age and creatine phosphokinase levels were significantly associated with a severe outcome. Seasonal respiratory infections are a continuing concern. Our results highlight the importance of studies on the prevalence and severity of these infections and that investments in programs of clinical and laboratory monitoring are essential to detect the appearance of new infective agents.
Development of a Cellular Assay System To Study the Genome Replication of High- and Low-Risk Mucosal and Cutaneous Human Papillomaviruses
Authors: Geimanen Jelizaveta , Isok-Paas Helen , Pipitch Regina , Salk Kristiina , Laos Triin , Orav Marit , Reinson Tormi , Ustav Mart , Ustav Mart , Ustav Ene 
Published: January 2011

ABSTRACT
We found that recircularized high-risk (type 16 and 18) and low-risk mucosal (type 6b and 11) and cutaneous (type 5 and 8) human papillomavirus (HPV) genomes replicate readily when delivered into U2OS cells by electroporation. The replication efficiency is dependent on the amount of input HPV DNA and can be followed for more than 3 weeks in proliferating cell culture without selection. Cotransfection of recircularized HPV genomes with a linear G418 resistance marker plasmid has allowed subcloning of cell lines, which, in a majority of cases, carry multicopy episomal HPV DNA. Analysis of the HPV DNA status in these established cell lines showed that HPV genomes exist in these cells as stable extrachromosomal oligomers. When the cell lines were cultivated as confluent cultures, a 3- to 10-fold amplification of the HPV genomes per cell was induced. Two-dimensional (2D) agarose gel electrophoresis confirmed amplification of mono- and oligomeric HPV genomes in these confluent cell cultures. Amplification occurred as a result of the initiation of semiconservative two-dimensional replication from one active origin in the HPV oligomer. Our data suggest that the system described here might be a valuable, cost-effective, and efficient tool for use in HPV DNA replication studies, as well as for the design of cell-based assays to identify potential inhibitors of all stages of HPV genome replication.
Requirement for the Amino-Terminal Domain of Sindbis Virus nsP4 during Virus Infection
Authors: Rupp Jonathan C. , Jundt Natasha , Hardy Richard W. 
Published: January 2011

ABSTRACT

The Sindbis virus RNA-dependent RNA polymerase nsP4 possesses an amino-terminal region that is unique to alphaviruses and is predicted to be disordered. To determine the importance of this region during alphavirus replication, 29 mutations were introduced, and resultant viruses were assessed for growth defects. Three small plaque mutants, D41A, G83L, and the triple mutant GPG
(8-10)
VAV, had defects in subgenome synthesis, minus-strand synthesis, and overall levels of viral RNA synthesis, respectively. Large plaque viruses were selected following passage in BHK-21 cells, and the genomes of these were sequenced. Suppressor mutations in nsP1, nsP2, and nsP3 that restored viral RNA synthesis were identified. An nsP2 change from M282 to L and an nsP3 change from H99 to N corrected the D41A-induced defect in subgenomic RNA synthesis. Three changes in nsP1, I351 to V, I388 to V, or the previously identified change, N374 to H (C. L. Fata, S. G. Sawicki, and D. L. Sawicki, J. Virol. 76:8641-8649, 2002), suppressed the minus-strand synthetic defect. A direct reversion back to G at position 8 reduced the RNA synthesis defect of the GPG
(8-10)
VAV virus. These results imply that nsP4's amino-terminal domain participates in distinct interactions with other nsPs in the context of differentially functioning RNA synthetic complexes, and flexibility in this domain is important for viral RNA synthesis. Additionally, the inability of the mutant viruses to efficiently inhibit host protein synthesis suggests a role for nsP4 in the regulation of host cell gene expression.
Prion Propagation in Cells Expressing PrP Glycosylation Mutants
Authors: Salamat Muhammad K. , Dron Michel , Chapuis Jérôme , Langevin Christelle , Laude Hubert 
Published: January 2011

ABSTRACT

Infection by prions involves conversion of a host-encoded cell surface protein (PrP
C
) to a disease-related isoform (PrP
Sc
). PrP
C
carries two glycosylation sites variably occupied by complex N-glycans, which have been suggested by previous studies to influence the susceptibility to these diseases and to determine characteristics of prion strains. We used the Rov cell system, which is susceptible to sheep prions, to generate a series of PrP
C
glycosylation mutants with mutations at one or both attachment sites. We examined their subcellular trafficking and ability to convert into PrP
Sc
and to sustain stable prion propagation in the absence of wild-type PrP. The susceptibility to infection of mutants monoglycosylated at either site differed dramatically depending on the amino acid substitution. Aglycosylated double mutants showed overaccumulation in the Golgi compartment and failed to be infected. Introduction of an ectopic glycosylation site near the N terminus fully restored cell surface expression of PrP but not convertibility into PrP
Sc
, while PrP
C
with three glycosylation sites conferred cell permissiveness to infection similarly to the wild type. In contrast, predominantly aglycosylated molecules with nonmutated N-glycosylation sequons, produced in cells expressing glycosylphosphatidylinositol-anchorless PrP
C
, were able to form infectious PrP
Sc
. Together our findings suggest that glycosylation is important for efficient trafficking of anchored PrP to the cell surface and sustained prion propagation. However, properly trafficked glycosylation mutants were not necessarily prone to conversion, thus making it difficult in such studies to discern whether the amino acid changes or glycan chain removal most influences the permissiveness to prion infection.
The V1V2 Domain and an N-Linked Glycosylation Site in the V3 Loop of the HIV-1 Envelope Glycoprotein Modulate Neutralization Sensitivity to the Human Broadly Neutralizing Antibody 2G12
Authors: Chaillon Antoine , Braibant Martine , Moreau Thierry , Thenin Suzie , Moreau Alain , Autran Brigitte , Barin Francis 
Published: January 2011

ABSTRACT

The broadly neutralizing human monoclonal antibody 2G12 binds to a carbohydrate-dependent epitope involving three major potential N-linked glycosylation sites (PNGS) of gp120 (N295, N332, and N392). Through analysis of the sensitivity to 2G12 of pseudotyped viruses carrying envelope proteins from HIV-1 clade B-infected long-term nonprogressors, we selected two naturally occurring
env
clones with opposite sensitivities to
2G12
, albeit harboring the 3 particular PNGS known to be essential for 2G12 binding (N295, N332, and N392). The resistant clone presented a long and potentially heavily glycosylated V1V2 loop and an additional PNGS (N302) in the V3 loop. The sensitive clone harbored a short V1V2 loop and lacked the PNGS at N302. We created chimeric envelope genes by swapping the V1V2 domains of the two
env
clones. The influence of N302 on 2G12 sensitivity was assessed by PCR-based site-directed mutagenesis. Both the exchange of the V1V2 domain and the introduction of the PNGS at N302 on the 2G12-sensitive clone induced a significant decrease in sensitivity to 2G12. In contrast, the reverse V1V2 exchange and the removal of the PNGS at N302 on the 2G12-resistant clone increased sensitivity to 2G12, confirming the influence of these regions on 2G12 sensitivity. Our results, supported by a molecular-modeling study, suggest that both the V1V2 loop and an additional PNGS in V3 might limit access to the 2G12 epitope.
Expression of the Human Endogenous Retrovirus (HERV) Group HML-2/HERV-K Does Not Depend on Canonical Promoter Elements but Is Regulated by Transcription Factors Sp1 and Sp3
Authors: Fuchs Nina V. , Kraft Martin , Tondera Christiane , Hanschmann Kay-Martin , Löwer Johannes , Löwer Roswitha 
Published: January 2011

ABSTRACT

After fixation in the human genome, human endogenous retroviruses (HERVs) are
bona fide
cellular genes despite their exogenous origin. To be able to spread within the germ line and the early embryo, the ancient retroviral promoters must have adapted to the requirements for expression in these cell types. We describe that in contrast to the case for current exogenous retroviruses, which replicate in specific somatic cells, the long terminal repeat (LTR) of the human endogenous retrovirus HERV-K acts as a TATA- and initiator element-independent promoter with a variable transcription start site. We present evidence that the HERV-K LTR is regulated by the transcription factors Sp1 and Sp3. Mutating specific GC boxes, which are binding sites for Sp proteins, and knocking down Sp1 and Sp3 by use of small interfering RNA (siRNA) significantly reduced the promoter activity. Binding of Sp1 and Sp3 to the promoter region was confirmed using electrophoretic mobility shift assays (EMSAs) and chromatin immunoprecipitation (ChIP). Our data explain why certain HERV-K proviruses have lost promoter competence. Since vertebrate promoters lacking canonical core promoter elements are common but poorly studied, understanding the HERV-K promoter not only will provide insight into the regulation of endogenous retroviruses but also can serve as a paradigm for understanding the regulation of this class of cellular genes.
Effect of anabolic steroid, IGF-1 and loading on expression of genes involved in collagen metabolism in bovine extensor tendon
Authors: Bhullar Sukhbir , Legerlotz Kirsten 
Published: January 2011

BackgroundTendon ruptures have been linked to anabolic steroid usage, suggesting pathological changes in tendon structure due to steroid intake. Anecdotal reports from athletes suggest a protective effect of GH on tendon, especially in those who use anabolic steroids.AimTo investigate how administration of insulin-like growth factor 1 (IGF-1), anabolic steroid and combination of the two affect the expression of genes involved in collagen metabolism.MethodsFascicles were harvested from bovine extensor tendons: n=4 to calculate optimal loading and n=8 to assess hormone plus optimal loading effect on eight genes involved in collagen metabolism. Total RNA was extracted and mRNA expression analysed by quantitative real-time reverse transcription-PCR, normalised to GAPDH and 18S ribosomal RNA±control (Con).ResultsMaximal collagen expression occurred at 6 h of light exercise, following 1 h of heavy loading. One tendon failed to have RNA extracted and was therefore removed from the study (n=7). IL-6 expression was significantly higher than control when exercised fascicles were exposed to: (1) anabolic steroid (normalised to: 18S; GAPDH; 18S & Con and GAPDH & Con); (2) IGF-1 (normalised to: GAPDH; 18S & Con and GAPDH & Con); (3) Dulbecco’s modified Eagle’s medium & PenStrep (normalised to: 18S & Con and GAPDH & Con) and (4) anabolic steroid & IGF-1 (normalised to: 18S & Con and GAPDH & Con).ConclusionThe results of this study suggest that tendon is an IL-6 producing region during exercise which may also contribute to IL-6 increases in plasma during exercise.
Longitudinal development of physiological characteristics in elite young males
Authors: Hemmings Steph , Nevill Mary , Morris John , Barrett Laura 
Published: January 2011

BackgroundCross-sectional studies examining training effects on development of performance variables have been confounded by growth and development changes. In order to examine the relative contribution of training, there was a need to collect ‘growth’ data longitudinally.Aims and objectivesThe purpose of this study was to examine age- and maturity-associated variation in longitudinal physiological characteristics development in elite and non-elite adolescent males.MethodsLongitudinal measures of peak VO2, submaximal VO2, and power output in 11 elite, (13.7–17.8 years), and five non-elite males (13.1–17.6 years), over 2 or 3 consecutive years were examined. Maturity was assessed using Tanner’s (1962) indices of secondary sexual characteristics.ResultsSum of skinfolds remained the same in the elite group yet increased in the non-elite. Peak VO2was greater in the elite males by 9.1 ml/kg/min, with no change in age. Oxygen uptake at 10.6 km/h declined by 1.2 ml/kg/min per year in each group. Percentage peak VO2at the same running speed was l2% lower in the elite and declined by 1.3% each year in both groups. Blood lactate concentration was 1.5 mmol/l lower in the elite males compared to the non-elite, however there was no decline with age. Peak and mean power output increased disproportionately in the elite group, with percentage fatigue increasing similarly.ConclusionBody fatness increases across adolescence in the untrained male. Peak VO2remains stable throughout adolescence but is characteristically higher in elites either as a result of strenuous training or genetic endowment. Running economy improves with age in both elite and non-elite but does not appear to be related to training status. Short-term power output shows a disproportionate increase with age in the elite male athlete compared to the untrained contemporary, which may have been influenced by differences in body composition between groups.
Variability of kinematic and kinetic gait data in ambulatory children with spastic cerebral palsy with and without fixed ankle-foot orthoses using 3d motion analysis: a quantitative prospective study
Authors: Mohammed U , Twycross-Lewis R , Timotijevic T , Woledge R , Bader D , Paterson M , Coggings D , Morrissey D 
Published: January 2011

BackgroundSpastic cerebral palsy (SCP) is a common neurological disorder causing inadequate motor-control and functional gait pattern. Fixed ankle-foot orthoses (FAFOs) have become a widespread conservative therapy in SCP. Gait of these children exhibits a large degree of within-child variability but few studies have investigated this variability in terms of kinematics and kinetics and no studies address how this variability is affected with the use of FAFOs.ObjectivesTo compare intrapatient variability of kinematic and kinetic gait parameters between a sample of ambulatory children with SCP and two healthy children with normal gait. The secondary aim was to assess if these variability profiles change in the SCP children as a result of FAFO application.MethodsFour SCP and two normal ambulatory children, aged 5–15, were recruited and underwent active instrumented gait analysis over a 6 week period. Variability of a measure was defined as the SD of the measure within one child over several repeated gait trials.ResultsVariability of sagittal-plane pelvic, hip, knee and ankle range of motion (all p<0.01) and sagittal internal hip and knee extensor moments (both p<0.05) were shown to be consistently greater in SCP than that of age-matched children. AFO application reduces sagittal kinematic variability at the pelvis, knee and ankle (all p<0.05) and variability of sagittal hip and knee moments (both p<0.01).ConclusionsChildren with SCP exhibit significantly higher kinematic and kinetic variability compared to the normal population. FAFOs have the potential to reduce this variability and functionally improve gait. Future studies should utilise larger sample sizes and stricter inclusion criteria taking into account the differing clinical severities of spasticity, FAFO designs and patterns of CP involvement to ensure these are generalisable findings.