bluuebunny/crossref_metadata_2025_split huggingface dataset:
Analysis of the IgG Immune Response toTreponema phagedenis-Like Spirochetes in Individual Dairy Cattle with Papillomatous Digital Dermatitis
Authors: Moe Kyaw Kyaw , Yano Takahisa , Misumi Kazuhiro , Kubota Chikara , Yamazaki Wataru , Muguruma Michio , Misawa Naoaki 
Published: January 2010

ABSTRACTPapillomatous digital dermatitis (PDD) is a major infectious disease of the foot skin in dairy cattle.Treponema phagedenis-like spirochetes have been consistently detected in PDD lesions, and antibodies against these organisms have been demonstrated in affected cattle. However, little is known about the dominant antigens recognized by the immune system of affected cattle. Here, we investigated the IgG immune response toT. phagedenis-like isolates by Western blotting with different sera using whole-cell lysates and extracted glycolipid from 18 and 8 isolates, respectively, including those from different cattle on the same or different farms, isolates from different lesions affecting a single cow, and different isolates from the same lesion affecting a single cow. The reactivity of sera in Western blot assays revealed different banding patterns or showed no bands, suggesting that considerable antigenic variations, including glycolipid, may exist among the isolates, even in those from single individuals. With use of a total of 151 serum samples collected from three groups of cattle, i.e., PDD-positive cows on PDD-positive farms (group A), PDD-negative cows on PDD-positive farms (group B), and cows on PDD-free farms (group C), the levels of IgG antibodies against fourT. phagedenis-like isolates were measured by enzyme-linked immunosorbent assay (ELISA). The optical density in groups A and B was significantly higher than that in group C, even though the value varied among the antigens used. Therefore, combinations of multipleTreponemaspecies should be used for serological analysis and the development of a suitable vaccine because of antigenic variations.
Novel Method for Genotyping Hepatitis B Virus on the Basis of TaqMan Real-Time PCR
Authors: Malmström Sebastian , Berglin-Enquist Ida , Lindh Magnus 
Published: January 2010

ABSTRACT
Chronic infection with hepatitis B virus (HBV) is an important cause of cirrhosis and cancer of the liver. HBV is currently classified into eight genotypes, A to H. Accumulated evidence shows that the genotype influences both the clinical course of infection and the response to treatment. We describe a new method for genotyping based on TaqMan real-time PCR, which identifies all HBV genotypes without post-PCR processing. In this assay, each sample is processed in four multiplex real-time PCRs, each targeting two or three genotype-specific segments of HBV. By analyzing 185 samples representing all genotypes and different proportions of genotype mixtures, we could validate high accuracy of the assay. We conclude that this new assay represents a significant advancement for both diagnostics and clinical research because it is accurate, practical, and based on a technique that is well established in many virological laboratories.
Molecular Phylogenetic Diversity of Dermatologic and Other Human Pathogenic Fusarial Isolates from Hospitals in Northern and Central Italy
Authors: Migheli Quirico , Balmas Virgilio , Harak Henry , Sanna Silvana , Scherm Barbara , Aoki Takayuki , O'Donnell Kerry 
Published: January 2010

ABSTRACTFifty-eight fusaria isolated from 50 Italian patients between 2004 and 2007 were subject to multilocus DNA sequence typing to characterize the spectrum of species and circulating sequence types (STs) associated with dermatological infections, especially onychomycoses and paronychia, and other fusarioses in northern and central Italy. Sequence typing revealed that the isolates were nearly evenly divided among theFusarium solanispecies complex (FSSC;n= 18), theF. oxysporumspecies complex (FOSC;n= 20), and theGibberella(Fusarium)fujikuroispecies complex (GFSC;n= 20). The three-locus typing scheme used for members of the FSSC identified 18 novel STs distributed among six phylogenetically distinct species, yielding an index of discrimination of 1.0. Phylogenetic analysis of the FOSC two-locus data set identified nine STs, including four which were novel, and nine isolates of ST 33, the previously described widespread clonal lineage. With the inclusion of eight epidemiologically unrelated ST 33 isolates, the FOSC typing scheme scored a discrimination index of 0.787. The two-locus GFSC typing scheme, which was primarily designed to identify species, received the lowest discrimination index, with a score of 0.492. The GFSC scheme, however, was used to successfully identify 17 isolates asF. verticillioides, 2 asF. sacchari, and 1 asF. guttiforme. This is the first report thatF. guttiformecauses a human mycotic infection, which was supported by detailed morphological analysis. In addition, the results of a pathogenicity experiment revealed that the human isolate ofF. guttiformewas able to induce fusariosis of pineapple, heretofore its only known host.
Hepatitis E Virus (HEV) Strains in Serum Samples Can Replicate Efficiently in Cultured Cells Despite the Coexistence of HEV Antibodies: Characterization of HEV Virions in Blood Circulation
Authors: Takahashi Masaharu , Tanaka Toshinori , Takahashi Hideyuki , Hoshino Yu , Nagashima Shigeo , Jirintai  , Mizuo Hitoshi , Yazaki Yasuyuki , Takagi Tomofumi , Azuma Masahiro , Kusano Eiji , Isoda Norio , Sugano Kentaro , Okamoto Hiroaki 
Published: January 2010

ABSTRACTWe recently developed a cell culture system for hepatitis E virus (HEV) in PLC/PRF/5 and A549 cells, using fecal specimens from HEV-infected patients. Since transfusion-associated hepatitis E has been reported, we examined PLC/PRF/5 and A549 cells for the ability to support replication of HEV in various serum samples obtained from 23 patients with genotype 1, 3, or 4 HEV. HEV progenies emerged in culture media of PLC/PRF/5 cells, regardless of the coexistence of HEV antibodies in serum but dependent on the load of HEV inoculated (31% at 2.0 × 104copies per well and 100% at ≥3.5 × 104copies per well), and were successfully passaged in A549 cells. HEV particles in serum, with or without HEV antibodies, banded at a sucrose density of 1.15 to 1.16 g/ml, which was markedly lower than that for HEV particles in feces, at 1.27 to 1.28 g/ml, and were nonneutralizable by immune sera in this cell culture system. An immuno-capture PCR assay of HEV virions treated with or without detergent indicated that HEV particles in serum are associated with lipids and HEV ORF3 protein, similar to those in culture supernatant. By immunoprecipitation, it was found that >90% of HEV particles in the circulation exist as free virions not complexed with immunoglobulins, despite the coexistence of HEV antibodies. These results suggest that ourin vitrocell culture system can be used for propagation of a wide variety of HEV strains in sera from various infected patients, allowing extended studies on viral replication specific to different HEV strains.
Combined Real-Time PCR and
rpoB
Gene Pyrosequencing for Rapid Identification of
Mycobacterium tuberculosis
and Determination of Rifampin Resistance Directly in Clinical Specimens
Authors: Halse Tanya A. , Edwards Justine , Cunningham Phyllis L. , Wolfgang William J. , Dumas Nellie B. , Escuyer Vincent E. , Musser Kimberlee A. 
Published: January 2010

ABSTRACT

Our laboratory has developed a rapid, sensitive, and specific molecular approach for detection in clinical specimens, within 48 h of receipt, of both
Mycobacterium tuberculosis
complex (MTBC) DNA and mutations within the 81-bp core region of the
rpoB
gene that are associated with rifampin (RIF) resistance. This approach, which combines an initial real-time PCR with internal inhibition assessment and a pyrosequencing assay, was validated for direct use with clinical specimens. To assess the suitability of real-time PCR for use with respiratory, nonrespiratory, acid-fast bacillus (AFB)-positive and AFB-negative specimens, we evaluated specimens received in our laboratory between 11 October 2007 and 30 June 2009. With culture used as the “gold standard,” the sensitivity, specificity, and positive and negative predictive values were determined for 1,316 specimens to be as follows: for respiratory specimens, 94.7%, 99.9%, 99.6%, and 98.6%, respectively; for nonrespiratory specimens, 88.5%, 100.0%, 100.0%, and 96.9%, respectively; for AFB-positive specimens, 99.6%, 100.0%, 100.0%, and 97.7%, respectively; and for AFB-negative specimens, 75.4%, 99.9%, 98.0%, and 98.4%, respectively. PCR inhibition was determined to be minimal in this assay, occurring in 0.2% of tests. The
rpoB
gene pyrosequencing assay was evaluated in a similar prospective study, in which 148 clinical specimens positive for MTBC DNA by real-time PCR were tested. The final results revealed that the results of direct testing of clinical specimens by the pyrosequencing assay were 98.6% concordant with the results of conventional testing for susceptibility to RIF in liquid culture and that our assay displayed adequate sensitivity for 96.6% of the clinical specimens tested. Used together, these assays provide reliable results that aid with the initial management of patients with suspected tuberculosis prior to the availability of the results for cultured material, and they also provide the ability to predict RIF resistance in
Mycobacterium tuberculosis
-positive specimens in as little as 48 h from the time of clinical specimen receipt.
Rapid Detection of
Staphylococcus aureus
Panton-Valentine Leukocidin in Clinical Specimens by Enzyme-Linked Immunosorbent Assay and Immunochromatographic Tests
Authors: Badiou Cedric , Dumitrescu Oana , George Narelle , Forbes Andrea R. N. , Drougka Eleanna , Chan Kian Sing , Ramdani-Bouguessa Nadjia , Meugnier Helene , Bes Michele , Vandenesch Francois , Etienne Jerome , Hsu Li Yang , Tazir Mohamed , Spiliopoulou Iris , Nimmo Graeme R. , Hulten Kristina G. , Lina Gerard 
Published: February 2010

ABSTRACT

Staphylococcus aureus
strains producing Panton-Valentine leukocidin (PVL) have been epidemiologically linked to specific human infections. To evaluate immunological tests that may be used to diagnose infections with PVL-producing strains, we prospectively collected pus, respiratory tract specimens, and joint fluid specimens from which
S. aureus
had been isolated in clinical laboratories in six countries. An enzyme-linked immunosorbent assay (ELISA) and an immunochromatographic test (ICT) targeting LukS-PV were performed directly with clinical samples for the detection of PVL. The same tests were applied to
S. aureus
culture supernatants. The corresponding
S. aureus
isolates were characterized by PCR for the presence of the PVL locus (
lukS-PV
and
lukF-PV
) and the
mec
A
gene. A total of 185 samples from 144 skin infections, 23 bone and joint infections, and 18 lower respiratory tract infections were analyzed. By PCR, 72/185
S. aureus
isolates were PVL locus positive (PVL
+
); 28 of these were also
mecA
positive. PVL was detected in the supernatants of all PVL
+
strains by both ELISA and an ICT, while no signal was observed with PVL-negative strains. The PVL concentrations in human clinical samples that grew PVL
+
strains ranged from 0 to 399 μg/ml by ELISA. By the use of 0.015 μg/ml of PVL as a cutoff value, PVL was detected in 65/72 (90%) of the clinical samples by ELISA. The sensitivity and specificity of the ELISA test were 90% and 100%, respectively. By the ICT, PVL was detected in 57/72 (79%) of the samples, and the sensitivity and specificity of ICT were 79% and 100%, respectively. PVL is expressed by
S. aureus
during human infection, and a PVL-specific ELISA and ICT could be reliable tests for the diagnosis of infections caused by PVL-producing strains.
Prevalence of Non-Penicillin-Susceptible Group B Streptococcus in Philadelphia and Specificity of Penicillin Resistance Screening Methods
Authors: Kasahara Kei , Baltus Andrew J. , Lee Sun-Hwa , Edelstein Martha A. , Edelstein Paul H. 
Published: January 2010

ABSTRACT
A total of 1,991 group B streptococcus (GBS) isolates, collected in 2008 and 2009, were tested for non-penicillin susceptibility by broth microdilution, disk testing, and oxacillin screening agar. No GBS isolates were non-penicillin susceptible. Oxacillin and ceftizoxime disk testing results showed that proposed screening criteria are nonspecific. The oxacillin screening agar was specific but of unknown sensitivity.
Stable Formation of Compositionally Unique Stress Granules in Virus-Infected Cells
Authors: Piotrowska Joanna , Hansen Spencer J. , Park Nogi , Jamka Katarzyna , Sarnow Peter , Gustin Kurt E. 
Published: January 2010

ABSTRACT

Stress granules are sites of mRNA storage formed in response to a variety of stresses, including viral infections. Here, the mechanisms and consequences of stress granule formation during poliovirus infection were examined. The results indicate that stress granules containing T-cell-restricted intracellular antigen 1 (TIA-1) and mRNA are stably constituted in infected cells despite lacking intact RasGAP SH3-domain binding protein 1 (G3BP) and eukaryotic initiation factor 4G. Fluorescent
in situ
hybridization revealed that stress granules in infected cells do not contain significant amounts of viral positive-strand RNA. Infection does not prevent stress granule formation in response to heat shock, indicating that poliovirus does not block
de novo
stress granule formation. A mutant TIA-1 protein that prevents stress granule formation during oxidative stress also prevents formation in infected cells. However, stress granule formation during infection is more dependent upon ongoing transcription than is formation during oxidative stress or heat shock. Furthermore, Sam68 is recruited to stress granules in infected cells but not to stress granules formed in response to oxidative stress or heat shock. These results demonstrate that stress granule formation in poliovirus-infected cells utilizes a transcription-dependent pathway that results in the appearance of stable, compositionally unique stress granules.
Characterization of the Varicella-Zoster Virus ORF50 Gene, Which Encodes Glycoprotein M
Authors: Sadaoka Tomohiko , Yanagi Tatsuya , Yamanishi Koichi , Mori Yasuko 
Published: January 2010

ABSTRACT

The ORF50 gene of the varicella-zoster virus (VZV) encodes glycoprotein M (gM), which is conserved among all herpesviruses and is important for the cell-to-cell spread of VZV. However, few analyses of ORF50 gene expression or its posttranscriptional and translational modifications have been published. Here we found that in VZV-infected cells, ORF50 encoded four transcripts: a full-size transcript, which was translated into the gM, and three alternatively spliced transcripts, which were not translated. Using a splicing-negative mutant virus, we showed that the alternative transcripts were nonessential for viral growth in cell culture. In addition, we found that two amino acid mutations of gM, V42P and G301M, blocked gM's maturation and transport to the
trans
-Golgi network, which is generally recognized as the viral assembly complex. We also found that the mutations disrupted gM's interaction with glycoprotein N (gN), revealing their interaction through a bond that is otherwise unreported for herpesviruses. Using this gM maturation-negative virus, we found that immature gM and gN were incorporated into intracellularly isolated virus particles and that mature gM was required for efficient viral growth via cell-to-cell spread but not for virion morphogenesis. The virus particles were more abundant at the abnormally enlarged perinuclear cisternae than those of the parental virus, but they were also found at the cell surface and in the culture medium. Additionally, in the gM maturation-negative mutant virus-infected melanoma cells, typical syncytium formation was rarely seen, again indicating that mature gM functions in cell-to-cell spread via enhancement of syncytium formation.
The ORF3 Protein of Hepatitis E Virus Delays Degradation of Activated Growth Factor Receptors by Interacting with CIN85 and Blocking Formation of the Cbl-CIN85 Complex
Authors: Chandra Vivek , Kalia Manjula , Hajela Krishnan , Jameel Shahid 
Published: February 2010

ABSTRACT

Hepatitis E virus (HEV) causes an acute self-limiting disease that is endemic in developing countries. Previous studies suggested that the ORF3 protein (pORF3) of HEV is required for infection
in vivo
and is likely to modulate the host response. Our previous work showed that pORF3 localizes to early and recycling endosomes and causes a delay in the postinternalization trafficking of epidermal growth factor receptor (EGFR) to late endosomes/lysosomes. Here we report that pORF3 also delays the trafficking and degradation of activated hepatocyte growth factor receptor (c-Met) and delineate the mechanistic details of these effects. A mutant ORF3 protein, which does not localize to endosomes, also showed similar effects on growth factor receptor trafficking, making this effect independent of the endosomal localization of pORF3. The ORF3 protein was found to interact with CIN85, a multidomain adaptor protein implicated in the Cbl-mediated downregulation of receptor tyrosine kinases. This interaction competed with the formation of the growth factor receptor-Cbl-CIN85 complex, resulting in the reduced ubiquitination of CIN85 and trafficking of the growth factor receptor complex toward late endosomes/lysosomes. We propose that through its effects on growth factor receptor trafficking, pORF3 prolongs endomembrane growth factor signaling and promotes cell survival to contribute positively to viral replication and pathogenesis.
Expression of Varicella-Zoster Virus Immediate-Early Regulatory Protein IE63 in Neurons of Latently Infected Human Sensory Ganglia
Authors: Zerboni Leigh , Sobel Raymond A. , Ramachandran Vasavi , Rajamani Jaya , Ruyechan William , Abendroth Allison , Arvin Ann 
Published: January 2010

ABSTRACT
Varicella-zoster virus (VZV) causes varicella and establishes latency in sensory nerve ganglia, but the characteristics of VZV latency are not well defined. Immunohistochemical detection of the VZV immediate-early 63 (IE63) protein in ganglion neurons has been described, but there are significant discrepancies in estimates of the frequency of IE63-positive neurons, varying from a rare event to abundant expression. We examined IE63 expression in cadaver ganglia using a high-potency rabbit anti-IE63 antibody and corresponding preimmune serum. Using standard immunohistochemical techniques, we evaluated 10 ganglia that contained VZV DNA from seven individuals. These experiments showed that neuronal pigments were a confounding variable; however, by examining sections coded to prevent investigator bias and applying statistical analysis, we determined that IE63 protein, if present, is in a very small proportion of neurons (<2.8%). To refine estimates of IE63 protein abundance, we modified our protocol by incorporating a biological stain to exclude the pigment signal and evaluated 27 ganglia from 18 individuals. We identified IE63 protein in neurons within only one ganglion, in which VZV glycoprotein E and an immune cell infiltrate were also demonstrated. Antigen preservation was shown by detection of neuronal synaptophysin. These data provide evidence that the expression of IE63 protein, which has been referred to as a latency-associated protein, is rare. Refining estimates of VZV protein expression in neurons is important for developing a hypothesis about the mechanisms by which VZV latency may be maintained.
CODING PARTITIONS OF REGULAR SETS
Authors: BÉAL MARIE-PIERRE , BURDERI FABIO , RESTIVO ANTONIO 
Published: January 2010

A coding partition of a set of words partitions this set into classes such that whenever a sequence, of minimal length, has two distinct factorizations, the words of these factorizations belong to the same class. The canonical coding partition is the finest coding partition that partitions the set of words in at most one unambiguous class and other classes that localize the ambiguities in the factorizations of finite sequences. We prove that the canonical coding partition of a regular set contains a finite number of regular classes and we give an algorithm for computing this partition. From this we derive a canonical decomposition of a regular monoid into a free product of finitely many regular monoids.
Fixed Point Theorems on Spaces Endowed with Vector-Valued Metrics
Authors: Filip Alexandru-Darius , Petruşel Adrian 
Published: January 2010

AbstractThe purpose of this work is to present some (local and global) fixed point results for singlevalued and multivalued generalized contractions on spaces endowed with vector-valued metrics. The results are extensions of some theorems given by Perov (1964), Bucur et al. (2009), M. Berinde and V. Berinde (2007), O'Regan et al. (2007), and so forth.
Platelet P2Y12 receptor antagonist pharmacokinetics and pharmaco -dynamics: A foundation for distinguishing mechanisms of bleeding and anticipated risk for platelet-directed therapies
Authors: Becker Richard , Gurbel Paul 
Published: February 2010

SummaryThe platelet P2Y12 receptor is involved in all aspects of arterial thrombosis, including adhesion, activation, aggregation, secretion and development of a stable aggregate on which coagulation proteins can assemble and fibrin strands can mesh. Inhibition of the P2Y12 receptor has been shown convincingly to reduce cardiovascular events among patients with acute coronary syndromes (ACS) and in patients undergoing percutaneous intervention (PCI). Current studies are exploring whether there is a threshold of platelet aggregation below which only more bleeding occurs, without a concomitant reduction in clinical events. The following review considers the potential relevance of reversible and irreversible mechanisms of P2Y12 inhibition to bleeding risk, posing the question, “Is it not only how much but how a platelet P2Y12 receptor is inhibited that determines the attributable safety profile?”
A genetic variant in the gene encoding fibrinogen beta chain predicted development of hypertension in Chinese men
Authors: Ong Kwok Leung , Tso Annette , Cherny Stacey , Sham Pak Chung , Lam Karen , Jiang Chao Qiang , Thomas Neil , Lam Tai Hing , Cheung Bernard 
Published: February 2010

SummaryFibrinogen, a major determinant of blood viscosity, is an acute phase protein associated with cardiovascular disease. We studied the association of hypertension with single nucleotide polymorphisms (SNPs) in the gene encoding the fibrinogen β chain (FGB). Three tagging SNPs (rs1025154, rs4220 and rs1044291) were selected from the HapMap database on Han Chinese. Genotypes were determined in 1,294 unrelated subjects from the Hong Kong Cardiovascular Risk Factor Prevalence Study cohort. There were 199 hypertensive subjects at baseline. Among 1,095 subjects normotensive at baseline, 178 developed hyper-tension during a median follow-up period of 6.4 years. Among the three tagging SNPs, rs4220 showed significant association with hypertension at both baseline (odds ratio [OR]=1.49, p=0.004) and at follow-up (OR=1.32, p=0.013). The minor A allele of this SNP was associated with higher plasma fibrinogen level (β=0.144, p<0.001 at baseline and β=0.130, p<0.001 at follow-up). Among subjects normotensive at baseline, this SNP was also associated with the development of hyper-tension in men (OR=1.52, p=0.022), but not in women. The SNP rs4220 in FGB, which leads to the substitution of arginine by lysine at position 448, is independently associated with plasma fibrinogen level and hypertension in Hong Kong Chinese. This suggests a possible causal role of fibrinogen in hypertension development, especially in men.