bluuebunny/crossref_metadata_2025_split huggingface dataset:
SIMPLIFIED CAPACITANCE MONITORING FOR THE DETERMINATION OF CAMPYLOBACTER SPP. GROWTH RATES
Authors: AKINS E. DEANN , RULE PATRICIA L. , KEENER MICHELLE R. , SELLERS HOLLY S. , LINE J. ERIC , SIMMONS MUSTAFA , HARRISON MARK A. , HIETT KELLI L. 
Published: March 2009

ABSTRACT Capacitance monitoring is commonly used as an efficient means to generate growth curves of bacterial pathogens. However, the use of capacitance monitoring with Campylobacter spp. was previously determined to be difficult due to the complexity of the required media. We investigated capacitance monitoring using a simplified medium for the efficient and reproducible construction of growth curves for Campylobacter spp. It was determined that Campylobacter spp. should initially be propagated on Mueller Hinton plates in a microaerobic atmosphere (5% O2, 10% CO2, 85% N2) at 37C for 24 h, followed by transfer to Mueller Hinton biphasic cultures for 6 h at 37C in a microaerobic atmosphere. Serial dilutions of the Campylobacter spp. cultures should be used for inoculation of Bactometer wells that contain 1 mL Mueller Hinton broth supplemented with 0.1 M sodium pyruvate for the completion of Campylobacter spp. growth curves with the Bactometer.PRACTICAL APPLICATIONSGrowth rates can vary greatly among Campylobacter isolates; therefore, the rapid and reproducible methods described will prove useful in studies where the generation of growth curves is critical for subsequent experimental analyses.
Internet and World Wide Web‐Based Tools for Neuropathology Practice and Education
Authors: Fung Kar‐Ming , Tihan Tarik 
Published: March 2009

AbstractThe Internet and the World Wide Web (www) serve as a source of information and a communication network. Together they form a so‐called web or network that allows for transmission and dissemination of information in unprecedented speed, volume and detail. This article presents an overview of the current status of neuropathology content on the www. As well as considering the Internet as a resource for neuropathology practice, education and research, we also address the issue of quality assurance when evaluating Internet and www content. Four major categories of websites (archival, broker, news and blog) are discussed and resources relevant to neuropathology of each type are highlighted. We believe that our report and similar attempts can provide an opportunity to discuss appropriate and effective use of the Internet by the neuropathology community.
Digital Pathology: A Tool for 21st Century Neuropathology
Authors: Guzman Miguel , Judkins Alexander R. 
Published: March 2009

AbstractDigital pathology represents an electronic environment for performing pathologic analysis and managing the information associated with this activity. The technology to create and support digital pathology has largely developed over the last decade. The use of digital pathology tools is essential to adapt and lead in the rapidly changing environment of 21st century neuropathology. The utility of digital pathology has already been demonstrated by pathologists in several areas including consensus reviews, quality assurance (Q/A), tissue microarrays (TMAs), education and proficiency testing. These utilities notwithstanding, interface issues, storage and image formatting all present challenges to the integration of digital pathology into the neuropathology work environment. With continued technologic improvements, as well as the introduction of fluorescent side scanning and multispectral detection, future developments in digital pathology offer the promise of adding powerful analytic tools to the pathology work environment. The integration of digital pathology with biorepositories offers particular promise for neuropathologists engaged in tissue banking. The utilization of these tools will be essential for neuropathologists to continue as leaders in diagnostics, translational research and basic science in the 21st century.
Competitive Electrochemiluminescence Wash and No-Wash Immunoassays for Detection of Serum Antibodies to Smooth
Brucella
Strains
Authors: Thompson Iain , McGiven John , Sawyer Jason , Thirlwall Rachel , Commander Nicola , Stack Judy 
Published: March 2009

ABSTRACT

Brucellosis is a bacterial zoonotic disease of major global importance. Natural hosts for
Brucella
species include animals of economic significance, such as cattle and small ruminants. Controlling brucellosis in natural hosts by high-throughput serological testing followed by the slaughter of seropositive animals helps to prevent disease transmission. This study aimed to convert an existing competitive enzyme-linked immunosorbent assay (cELISA), used for the serodiagnosis of brucellosis in ruminants, to two electrochemiluminescence (ECL) immunoassays on the Meso Scale Discovery (MSD) platform. The first assay employed a conventional plate washing step as part of the protocol. The second was a no-wash assay, made possible by the proximity-based nature of ECL signal generation by the MSD platform. Both ECL wash and no-wash assays closely matched the parent cELISA for diagnostic sensitivity and specificity. The results also demonstrated that both ECL assays met World Organization for Animal Health (OIE) standards, as defined by results for the OIE standard serum (OIEELISA
SP
SS). This report is the first to describe an ECL assay incorporating lipopolysaccharide, an ECL assay for serodiagnosis of a bacterial infectious disease, a separation-free (no-wash) ECL assay for the detection of serum antibodies, and the use of the MSD platform for serodiagnosis. The simple conversion of the cELISA to the MSD platform suggests that many other serodiagnostic tests could readily be converted. Furthermore, the alignment of these results with the multiplex capability of the MSD platform offers the potential of no-wash multiplex assays to screen for several diseases.
Immunogenicity of a Whole-Cell Pertussis Vaccine with Low Lipopolysaccharide Content in Infants
Authors: Zorzeto Tatiane Queiroz , Higashi Hisako Gondo , da Silva Marcos Tadeu Nolasco , Carniel Emilia de Faria , Dias Waldely Oliveira , Ramalho Vanessa Domingues , Mazzola Taís Nitsch , Lima Simone Corte Batista Souza , Morcillo André Moreno , Stephano Marco Antonio , Antonio Maria Ângela Reis de Góes , Zanolli Maria de Lurdes , Raw Isaias , Vilela Maria Marluce dos Santos 
Published: March 2009

ABSTRACTThe lack of a clear correlation between the levels of antibody to pertussis antigens and protection against disease lends credence to the possibility that cell-mediated immunity provides primary protection against disease. This phase I comparative trial had the aim of comparing the in vitro cellular immune response and anti-pertussis toxin (anti-PT) immunoglobulin G (IgG) titers induced by a cellular pertussis vaccine with low lipopolysaccharide (LPS) content (wPlowvaccine) with those induced by the conventional whole-cell pertussis (wP) vaccine. A total of 234 infants were vaccinated at 2, 4, and 6 months with the conventional wP vaccine or the wPlowvaccine. Proliferation of CD3+T cells was evaluated by flow cytometry after 6 days of peripheral blood mononuclear cell culture with stimulation with heat-killedBordetella pertussisor phytohemagglutinin (PHA). CD3+, CD4+, CD8+, and T-cell receptor γδ-positive (γδ+) cells were identified in the gate of blast lymphocytes. Gamma interferon, tumor necrosis factor alpha, interleukin-4 (IL-4), and IL-10 levels in supernatants and serum anti-PT IgG levels were determined using enzyme-linked immunosorbent assay (ELISA). The net percentage of CD3+blasts in cultures withB. pertussisin the group vaccinated with wP was higher than that in the group vaccinated with the wPlowvaccine (medians of 6.2% for the wP vaccine and 3.9% for the wPlowvaccine;P= 0.029). The frequencies of proliferating CD4+, CD8+, and γδ+cells, cytokine concentrations in supernatants, and the geometric mean titers of anti-PT IgG were similar for the two vaccination groups. There was a significant difference between the T-cell subpopulations forB. pertussisand PHA cultures, with a higher percentage of γδ+cells in theB. pertussiscultures (P< 0.001). The overall data did suggest that wP vaccination resulted in modestly better specific CD3+cell proliferation, and γδ+cell expansions were similar with the two vaccines.
Rapid Mycobacterial Liquid Culture-Screening Method forMycobacterium aviumComplex Based on Secreted Antigen-Capture Enzyme-Linked Immunosorbent Assay
Authors: Shin Sung Jae , Anklam Kelly , Manning Elizabeth J. B. , Collins Michael T. 
Published: March 2009

ABSTRACTSensors in automated liquid culture systems for mycobacteria, such as MGIT, BacT/Alert 3D, and Trek ESP II, flag growth of any type of bacteria; a positive signal does not mean that the target mycobacteria are present. All signal-positive cultures thus require additional and often laborious testing. An immunoassay was developed to screen liquid mycobacterial cultures for evidence ofMycobacterium aviumcomplex (MAC). The method, called the MAC-enzyme-linked immunosorbent assay (ELISA), relies on detection of MAC-specific secreted antigens in liquid culture. Secreted MAC antigens were captured by the MAC-ELISA with polyclonal anti-Mycobacterium aviumsubsp.paratuberculosischicken immunoglobulin Y (IgY), detected using rabbit anti-MAC IgG, and then revealed using horseradish peroxidase-conjugated goat anti-rabbit IgG. When the MAC-ELISA was evaluated using pure cultures of known mycobacterial (n= 75) and nonmycobacterial (n= 17) organisms, no false-positive or false-negative MAC-ELISA results were found. By receiver operator characteristic (ROC) analysis of 1,275 previously identified clinical isolates, at the assay optimal cutoff the diagnostic sensitivity and specificity of the MAC-ELISA were 92.6% (95% confidence interval [95% CI], 90.3 to 94.5) and 99.9% (95% CI, 99.2 to 100), respectively, with an area under the ROC curve of 0.992. Prospective evaluation of the MAC-ELISA with an additional 652 clinical samples inoculated into MGIT ParaTB medium and signaling positive per the manufacturer's instructions found that the MAC-ELISA was effective in determining those cultures that actually contained MAC species and warranting the resources required to identify the organism by PCR. Of these 652 MGIT-positive cultures, the MAC-ELISA correctly identified 96.8% (of 219 MAC-ELISA-positive cultures) as truly containing MAC mycobacteria, based on PCR or high-performance liquid chromatography (HPLC) as reference tests. Only 6 of 433 MGIT signal-positive cultures (1.4%) were MAC-ELISA false negative, and only 7 of 219 MGIT signal-negative cultures (3.2%) were false positive. The MAC-ELISA is a low-cost, rapid, sensitive, and specific test for MAC in liquid cultures. It could be used in conjunction with or independent of automated culture reading instrumentation. For maximal accuracy and subspecies-specific identification, use of a confirmatory multiplex MAC PCR is recommended.
Antibody Response to Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Nonstructural Proteins and Implications for Diagnostic Detection and Differentiation of PRRSV Types I and II
Authors: Brown Elizabeth , Lawson Steven , Welbon Craig , Gnanandarajah Josephine , Li Juan , Murtaugh Michael P. , Nelson Eric A. , Molina Ramon M. , Zimmerman Jeffery J. , Rowland Raymond R. R. , Fang Ying 
Published: March 2009

ABSTRACT
To further characterize the humoral immune response of pigs to porcine reproductive and respiratory syndrome virus (PRRSV), direct enzyme-linked immunosorbent assays (ELISA) were used to study the kinetics of antibody responses directed against PRRSV nonstructural proteins in pigs experimentally exposed to the virus. The highest immunoreactivities were against nsp1, nsp2, and nsp7. Using the recombinant nsp7 as an antigen, we validated a dual ELISA for the simultaneous detection and differentiation of serum antibodies against type I and type II PRRSV. Receiver operating characteristic analysis based on 1,334 known-positive and 1,357 known-negative samples showed good specificity (98.3% to type I and 99.3% to type II) and sensitivity (97.4% for type I and 99.8% for type II). To differentiate type I and type II PRRSV, 470 sera originating from experimentally inoculated pigs were tested, and positive sera were correctly differentiated in 469 of 470 samples. The capability of the nsp7 dual ELISA to detect serum antibody responses from pigs infected with various genetically different field strains was determined. The nsp7 dual ELISA possessed 97.6% agreement with the Idexx HerdChek PRRS 2XR ELISA. In further testing of Idexx ELISA suspected false-positive samples, the nsp7 dual ELISA resolved 98% of the samples as negative. Taken together, these results indicate that the nsp7 dual ELISA can be used as a differential test for PRRSV serology with high levels of sensitivity and specificity. This ELISA offers an additional tool for routine or follow-up diagnostics, as well as having substantial value in epidemiological surveys and outbreak investigations.
Molecular Genetic Characterization of the Merozoite Surface Protein 1 Gene of
Plasmodium vivax
from Reemerging Korean Isolates
Authors: Kim So-Hee , Hwang Seung-Young , Shin Jeong Hwan , Moon Chi-Sook , Kim Dong-Wook , Kho Weon-Gyu 
Published: March 2009

ABSTRACT

Plasmodium vivax
merozoite surface protein 1 (PvMSP-1) has been considered a major candidate for the development of an antimalaria vaccine, but the molecule exhibits antigenic diversity among isolates. The extent of genetic polymorphism in the region between interspecies conserved blocks 4 and 5 (ICB4 and ICB5) of the PvMSP-1 gene was analyzed for 30 Korean isolates. Two genotypes, SK-A and SK-B, were identified on the basis of amino acid substitution. Almost all the amino acid sequences of the Korean isolates were nearly identical to those of the Solomon Island isolate Solo-83 (97.8 to 99.9% similarity) and Philippine isolates Ph-79, Ph-52-2, and Ph-49 (97.3 to 99.8% similarity). Also, we report two sequences in the isolates that were characterized on the basis of restriction fragment length polymorphism (RFLP). The RFLP profiles following digestion with the DraI restriction enzyme produced two distinguishable patterns. This study might be the first report of the region between ICB4 and ICB5 of the MSP-1 gene of
P. vivax
in South Korea.
Low Rates of
Pseudomonas aeruginosa
Misidentification in Isolates from Cystic Fibrosis Patients
Authors: Kidd Timothy J. , Ramsay Kay A. , Hu Honghua , Bye Peter T. P. , Elkins Mark R. , Grimwood Keith , Harbour Colin , Marks Guy B. , Nissen Michael D. , Robinson Philip J. , Rose Barbara R. , Sloots Theo P. , Wainwright Claire E. , Bell Scott C. 
Published: March 2009

ABSTRACT

Pseudomonas aeruginosa
is an important cause of pulmonary infection in cystic fibrosis (CF). Its correct identification ensures effective patient management and infection control strategies. However, little is known about how often CF sputum isolates are falsely identified as
P. aeruginosa
. We used
P. aeruginosa
-specific duplex real-time PCR assays to determine if 2,267
P. aeruginosa
sputum isolates from 561 CF patients were correctly identified by 17 Australian clinical microbiology laboratories. Misidentified isolates underwent further phenotypic tests, amplified rRNA gene restriction analysis, and partial 16S rRNA gene sequence analysis. Participating laboratories were surveyed on how they identified
P. aeruginosa
from CF sputum. Overall, 2,214 (97.7%) isolates from 531 (94.7%) CF patients were correctly identified as
P. aeruginosa
. Further testing with the API 20NE kit correctly identified only 34 (59%) of the misidentified isolates. Twelve (40%) patients had previously grown the misidentified species in their sputum.
Achromobacter xylosoxidans
(
n
= 21),
Stenotrophomonas maltophilia
(
n
= 15), and
Inquilinus limosus
(
n
= 4) were the species most commonly misidentified as
P. aeruginosa
. Overall, there were very low rates of
P. aeruginosa
misidentification among isolates from a broad cross section of Australian CF patients. Additional improvements are possible by undertaking a culture history review, noting colonial morphology, and performing stringent oxidase, DNase, and colistin susceptibility testing for all presumptive
P. aeruginosa
isolates. Isolates exhibiting atypical phenotypic features should be evaluated further by additional phenotypic or genotypic identification techniques.
Clostridium glycolicum
Wound Infections: Case Reports and Review of the Literature
Authors: Jiang Wei , Abrar Sahibzada , Romagnoli Mark , Carroll Karen C. 
Published: March 2009

ABSTRACT

We describe two cases of
Clostridium glycolicum
wound infections in immunocompetent adults. The bacterium was identified by 16S rRNA gene sequencing. This is the third published report of the recovery of this organism from human clinical material and highlights the importance of the organism as a potential human pathogen. Our report extends the spectrum of the diseases caused by
C. glycolicum
.
The putative tumour modifier gene ATP5A1 is not mutated in human colorectal cancer cell lines but expression levels correlate with TP53 mutations and chromosomal instability
Authors: Seth R , Keeley J , Abu-Ali G , Crook S , Jackson D , Ilyas M 
Published: March 2009

Background:Both the putative modifier gene ATP5a1 and the tumour suppressor gene TP53 are involved in the regulation of apoptosis and may be involved in the development of colorectal cancers.Aims:To investigate the relationship between these genes in 16 colorectal cancer cell lines.Methods:Each gene was screened for mutation using high resolution melting analysis and sequencing. Expression of ATP5a1 mRNA was tested by quantitative PCR.Results:Sequence changes in ATP5a1 were found in 9/16 (56%) cell lines and consisted of mainly novel single nucleotide polymorphisms (SNPs) found in the 5′ UTR, introns 4/5/9 and exon 7. TP53 mutations were also found in 9/16 (56%) cell lines; these were consistent with previous reports. High levels of ATP5a1 expression were seen in cell lines with TP53 mutation compared with those with wild type TP53 (p = 0.02). Furthermore, an A→G change at the −18 position in intron 4 of ATP5a1 was significantly associated with increased gene expression (p = 0.0391). Comparison with genotype showed that cell lines with chromosomal instability (CIN) had significantly higher levels of ATP5a1 expression than those with microsatellite instability (MSI) (p = 0.02).Conclusion:Higher levels of ATP5a1 expression are associated with certain SNPs and withTP53mutation. High expression also occurs in CIN and may facilitate tumour development along this pathway. Conversely, low levels of ATP5a1 expression may facilitate development of tumours with MSI.
Differential stepwise evolution of SARS coronavirus functional proteins in different host species
Authors: Tang Xianchun , Li Gang , Vasilakis Nikos , Zhang Yuan , Shi Zhengli , Zhong Yang , Wang Lin-Fa , Zhang Shuyi 
Published: March 2009

Abstract

Background
SARS coronavirus (SARS-CoV) was identified as the etiological agent of SARS, and extensive investigations indicated that it originated from an animal source (probably bats) and was recently introduced into the human population via wildlife animals from wet markets in southern China. Previous studies revealed that the spike (S) protein of SARS had experienced adaptive evolution, but whether other functional proteins of SARS have undergone adaptive evolution is not known.

Results
We employed several methods to investigate selective pressure among different SARS-CoV groups representing different epidemic periods and hosts. Our results suggest that most functional proteins of SARS-CoV have experienced a stepwise adaptive evolutionary pathway. Similar to previous studies, the spike protein underwent strong positive selection in the early and middle phases, and became stabilized in the late phase. In addition, the replicase experienced positive selection only in human patients, whereas assembly proteins experienced positive selection mainly in the middle and late phases. No positive selection was found in any proteins of bat SARS-like-CoV. Furthermore, specific amino acid sites that may be the targets of positive selection in each group are identified.

Conclusion
This extensive evolutionary analysis revealed the stepwise evolution of different functional proteins of SARS-CoVs at different epidemic stages and different hosts. These results support the hypothesis that SARS-CoV originated from bats and that the spill over into civets and humans were more recent events.
The outcome of Cryptococcus neoformansintracellular pathogenesis in human monocytes
Authors: Alvarez Mauricio , Burn Tamika , Luo Yong , Pirofski Liise-anne , Casadevall Arturo 
Published: March 2009

AbstractBackgroundCryptococcus neoformansis an encapsulated yeast that is a facultative intracellular pathogen. The interaction between macrophages andC. neoformansis critical for extrapulmonary dissemination of this pathogenic yeast.C. neoformanscan either lyse macrophages or escape from within them through a process known as phagosomal extrusion. However, most studies of intracellular pathogenesis have been made with mouse cells and their relevance to human infection is uncertain. In this study we extended studies ofC. neoformans-macrophage cellular interaction/s to human peripheral blood monocytes.ResultsThis study demonstrated thatC. neoformanscan shed polysaccharide within human monocytes, spread from cell to cell, and be extruded from them. Furthermore, human monocytes responded to ingestion ofC. neoformanswith cell cycle progression from G1 to S.ConclusionSimilarities between mouse and human cells support the suitability of mouse cells for the study of intracellular pathogenesis mechanisms. Given that these hosts diverged over 70 million years ago, the similar pathogenic strategies forC. neoformansin murine and human cells supports the hypothesis that the mechanism that underlies the mammalian intracellular pathogenesis ofC. neoformansoriginated from interactions with a third host, possibly soil amoeboid predators, before the mammalian radiation.
Zinc transporter gene expression is regulated by pro-inflammatory cytokines: a potential role for zinc transporters in beta-cell apoptosis?
Authors: Egefjord Lærke , Jensen Jens Ledet , Bang-Berthelsen Claus Heiner , Petersen Andreas Brønden , Smidt Kamille , Schmitz Ole , Karlsen Allan Ertman , Pociot Flemming , Chimienti Fabrice , Rungby Jørgen , Magnusson Nils E 
Published: March 2009

Abstract

Background
β-cells are extremely rich in zinc and zinc homeostasis is regulated by zinc transporter proteins. β-cells are sensitive to cytokines, interleukin-1β (IL-1β) has been associated with β-cell dysfunction and -death in both type 1 and type 2 diabetes. This study explores the regulation of zinc transporters following cytokine exposure.

Methods
The effects of cytokines IL-1β, interferon-γ (IFN-γ), and tumor necrosis factor-α (TNF-α) on zinc transporter gene expression were measured in INS-1-cells and rat pancreatic islets. Being the more sensitive transporter, we further explored ZnT8 (Slc30A8): the effect of ZnT8 over expression on cytokine induced apoptosis was investigated as well as expression of the insulin gene and two apoptosis associated genes, BAX and BCL2.

Results
Our results showed a dynamic response of genes responsible for β-cell zinc homeostasis to cytokines: IL-1β down regulated a number of zinc-transporters, most strikingly ZnT8 in both islets and INS-1 cells. The effect was even more pronounced when mixing the cytokines. TNF-α had little effect on zinc transporter expression. IFN-γ down regulated a number of zinc transporters. Insulin expression was down regulated by all cytokines. ZnT8 over expressing cells were more sensitive to IL-1β induced apoptosis whereas no differences were observed with IFN-γ, TNF-α, or a mixture of cytokines.

Conclusion
The zinc transporting system in β-cells is influenced by the exposure to cytokines. Particularly ZnT8, which has been associated with the development of diabetes, seems to be cytokine sensitive.