bluuebunny/crossref_metadata_2025 huggingface dataset:
Host Factors and Portal of Entry Outweigh Bacterial Determinants To Predict the Severity of Escherichia coli Bacteremia
Authors: Lefort Agnès , Panhard Xavière , Clermont Olivier , Woerther Paul-Louis , Branger Catherine , Mentré France , Fantin Bruno , Wolff Michel , Denamur Erick
Published: December 2010

Escherichia coli
ranks among the organisms most frequently isolated from cases of bacteremia. The relative contribution of the host and bacteria to
E. coli
bacteremia severity remains unknown. We conducted a prospective multicenter cohort study to identify host and bacterial factors associated with
E. coli
bacteremia severity. The primary endpoint was in-hospital death, up to 28 days after the first positive blood culture. Among 1,051 patients included, 136 (12.9%) died. Overall, 604 (57.5%) patients were female. The median age was 70 years, and 202 (19.2%) episodes were nosocomial. The most frequent comorbidities were immunocompromised status (37.9%), tobacco addiction (21.5%), and diabetes mellitus (20.1%). The most common portal of entry was the urinary tract (56.9%). Most
E. coli
isolates belonged to phylogenetic group B2 (52.0%). The multivariate analysis retained the following factors as predictive of death: older age (odds ratio [OR] = 1.25 [95% confidence interval {CI}, 1.09 to 1.43] for each 10-year increment), cirrhosis (OR = 4.85 [95% CI, 2.49 to 9.45]), hospitalization before bacteremia (OR = 4.13 [95% CI, 2.49 to 6.82]), being an immunocompromised patient not hospitalized before bacteremia (OR = 3.73 [95% CI, 2.25 to 6.18]), and a cutaneous portal of entry (OR = 6.45 [95% CI, 1.68 to 24.79]); a urinary tract portal of entry and the presence of the
ireA
virulence gene were negatively correlated with death (OR = 0.46 [95% CI, 0.30 to 0.70] and OR = 0.53 [95% CI, 0.30 to 0.91], respectively). In summary, host factors and the portal of entry outweigh bacterial determinants for predicting
E. coli
bacteremia severity.
Emergence of a Novel Shigella flexneri Serotype 4s Strain That Evolved from a Serotype X Variant in China
Authors: Qiu Shaofu , Wang Zhongqiang , Chen Chen , Liu Nan , Jia Leili , Liu Wenjing , Wang Ligui , Hao Rongzhang , Zhang Ling , Wang Yong , Song Hongbin
Published: December 2010

This paper describes the first isolation of a new
Shigella flexneri
serotype, designated 4s, in Beijing, China. Genotypic and phenotypic profiling suggests that this isolate is a clone of the
S. flexneri
serotype X variant reference strain. Of particular concern is the multidrug resistance exhibited by this isolate.
Evaluation of the 3M Rapid Detection Test for Respiratory Syncytial Virus (RSV) in Children during the Early Stages of the 2009 RSV Season
Authors: Munjal Iona , Gialanella Philip , Goss Cheryl , McKitrick John C. , Avner Jeffrey R. , Pan Qiulu , Litman Nathan , Levi Michael H.
Published: December 2010

We report the results of the 3M rapid detection respiratory syncytial virus (RSV) assay. This study includes pediatric patient results from nasopharyngeal swabs submitted from October to December 2009. There was a sensitivity of 74% and specificity approaching 100% compared to the PCR-based xTAG respiratory viral panel.
Development and Evaluation of a Reverse Transcription-Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Enterovirus 71
Authors: Jiang Tao , Liu Juan , Deng Yong-Qiang , Xu Li-Juan , Li Xiao-Feng , Han Jian-Feng , Cao Rui-Yuan , Qin E-De , Qin Cheng-Feng
Published: December 2010

Human enterovirus 71 (EV71) is the major etiological agent of hand, foot, and mouth disease (HFMD), which is a common infectious disease in young children and infants. EV71 can cause various clinical manifestations and has been associated with severe neurological complications; it has resulted in fatalities during recent outbreaks in Asian-Pacific regions since 1997. The early and rapid detection is critical for prevention and control of EV71 infection, since no vaccine or antiviral drugs are currently available. In this study, a simple and sensitive reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was developed for rapid detection of EV71. The detection limit of the RT-LAMP assay was approximately 0.01 PFU per reaction mixture, and no cross-reactive amplification with other enteroviruses was observed. The assay was evaluated further with 40 clinical specimens and exhibited 92.9% sensitivity and 100% specificity. This RT-LAMP assay may become a useful alternative in clinical diagnosis of EV71, especially in resource-limited hospitals or rural clinics of China and other countries in the Asian-Pacific region.
Distinct Host Species Correlate with
Anaplasma phagocytophilum ankA
Gene Clusters
Authors: Scharf Wiebke , Schauer Sonja , Freyburger Felix , Petrovec Miroslav , Schaarschmidt-Kiener Daniel , Liebisch Gabriele , Runge Martin , Ganter Martin , Kehl Alexandra , Dumler J. Stephen , Garcia-Perez Ana L. , Jensen Jennifer , Fingerle Volker , Meli Marina L. , Ensser Armin , Stuen Snorre , von Loewenich Friederike D.
Published: December 2010

Anaplasma phagocytophilum
is a Gram-negative, tick-transmitted, obligate intracellular bacterium that elicits acute febrile diseases in humans and domestic animals. In contrast to the United States, human granulocytic anaplasmosis seems to be a rare disease in Europe despite the initial recognition of
A. phagocytophilum
as the causative agent of tick-borne fever in European sheep and cattle. Considerable strain variation has been suggested to occur within this species, because isolates from humans and animals differed in their pathogenicity for heterologous hosts. In order to explain host preference and epidemiological diversity, molecular characterization of
A. phagocytophilum
strains has been undertaken. Most often the 16S rRNA gene was used, but it might be not informative enough to delineate distinct genotypes of
A. phagocytophilum
. Previously, we have shown that
A. phagocytophilum
strains infecting
Ixodes ricinus
ticks are highly diverse in their
ankA
genes. Therefore, we sequenced the 16S rRNA and
ankA
genes of 194
A. phagocytophilum
strains from humans and several animal species. Whereas the phylogenetic analysis using 16S rRNA gene sequences was not meaningful, we showed that distinct host species correlate with
A. phagocytophilum ankA
gene clusters.
Shiga Toxin-Producing Escherichia coli in Children: Diagnosis and Clinical Manifestations of O157:H7 and Non-O157:H7 Infection
Authors: Hermos Christina R. , Janineh Marcie , Han Linda L. , McAdam Alexander J.
Published: December 2010

Shiga toxin-producingEscherichia coli(STEC), a cause of food-borne colitis and hemolytic-uremic syndrome in children, can be serotype O157:H7 (O157) or other serotypes (non-O157).E. coliO157 can be detected by culture with sorbitol-MacConkey agar (SMAC), but non-O157 STEC cannot be detected with this medium. Both O157 and non-O157 STEC can be detected by immunoassay for Shiga toxins 1 and 2. The objectives of this study were first to compare the diagnostic utility of SMAC to that of the Premier EHEC enzyme immunoassay (Meridian Diagnostics) for detection of STEC in children and second to compare the clinical and laboratory characteristics of children with serotype O157:H7 STEC and non-O157:H7 STEC infections. Stool samples submitted for testing for STEC between April 2004 and September 2009 were tested by both SMAC culture and the Premier EHEC assay at Children's Hospital Boston. Samples positive by either test were sent for confirmatory testing and serotyping at the Hinton State Laboratory Institute (HSLI). Chart review was performed on children with confirmed STEC infection. Of 5,110 children tested for STEC, 50 (0.9%) had STEC infection confirmed by culture; 33 were O157:H7 and 17 were non-O157:H7. The Premier EHEC assay and SMAC culture detected 96.0% and 58.0% of culture-confirmed STEC isolates (any serotype), respectively, and 93.9% and 87.9% of STEC O157:H7 isolates, respectively. There were no significant differences in disease severity or laboratory manifestations of STEC infection between children with O157:H7 and those with non-O157 STEC. The Premier EHEC assay was significantly more sensitive than SMAC culture for diagnosis of STEC, and O157:H7 and non-O157:H7 STEC caused infections of similar severity in children.
High Frequency of Repeated Infections Due to Emerging Genotypes of Human Respiratory Syncytial Viruses among Children during Eight Successive Epidemic Seasons in Japan
Authors: Yamaguchi Masahiro , Sano Yasuko , Dapat Isolde C. , Saito Reiko , Suzuki Yasushi , Kumaki Akihiko , Shobugawa Yugo , Dapat Clyde , Uchiyama Makoto , Suzuki Hiroshi
Published: December 2010

In eight successive seasons (2001 to 2009), a total of 726 human respiratory syncytial virus (HRSV) infections from a total of 1,560 children with acute lower respiratory tract illness were identified. Molecular analysis of the attachment (G) protein gene confirmed that 52 (7.8%) children were infected more than once with any of the 3 genotypes of HRSV-A (genotypes GA5, NA1, and NA2) and/or 6 genotypes of HRSV-B (genotypes BA4, BA5, and BA7 to BA10). Repeated infections in 46 cases (82.1%) occurred in the next season, and only one case occurred in the same season (10-day interval). First infections were 33 (63.5%) HRSV-A cases and 19 (36.5%) HRSV-B cases, whereas second infections occurred in 35 (67.3%) HRSV-A cases and 17 (32.7%) HRSV-B cases. Third infections were attributed to 4 (100.0%) HRSV-A cases. Homologous subgroup reinfections were detected in 28 cases, 23 HRSV-A cases and 5 HRSV-B cases (
P
= 0.005), whereas homologous genotype reinfections were detected only for 5 HRSV-A cases (2GA5 and 3NA2) but not any HRSV-B case. Heterologous subgroup reinfections were detected in 28 cases, 12 cases from HRSV-A-to-HRSV-B reinfections and 16 cases from HRSV-B-to-HRSV-A reinfections. Genotypes NA1 and NA2 had higher numbers of heterologous genotype infections than did other genotypes. Our observations suggest that repeated infections occur more frequently in HRSV-A strains than in HRSV-B strains, and heterologous genotype reinfections occur more frequently than homologous genotype reinfections, especially in the case of the emerging genotypes NA1 and NA2 of HRSV-A strains that circulated in the community during our study period.
Characteristics of Cryptosporidium Transmission in Preweaned Dairy Cattle in Henan, China
Authors: Wang Rongjun , Wang Helei , Sun Yanru , Zhang Longxian , Jian Fuchun , Qi Meng , Ning Changshen , Xiao Lihua
Published: December 2010

To estimate the prevalence and public health significance of cryptosporidiosis in preweaned calves in China, 801 fecal samples from eight farms in seven areas in Henan Province were examined for
Cryptosporidium
oocysts. The overall infection rate of
Cryptosporidium
was 21.5%, with the farm in Xinxiang having the highest prevalence (40%). No significant difference in infection rates was observed between seasons.
Cryptosporidium
spp. were characterized by PCR-restriction fragment length polymorphism (RFLP) analysis of the small subunit (SSU) rRNA gene and DNA sequencing of the 60-kDa glycoprotein (gp60) gene. The SSU rRNA-based PCR identified four
Cryptosporidium
species, including
Cryptosporidium parvum
(54/172),
C. bovis
(65/172),
C. ryanae
(19/172), and
C. andersoni
(12/172), and the occurrence of infections with mixed species (22/172). The earliest detection of
C. bovis
was in calves of 1 week of age, showing that the prepatent period was shorter than the previously stated 10 to 12 days. Infections with
C. parvum
peaked in summer, whereas
C. bovis
dominated in autumn and winter. There was no apparent difference in the age of cattle infected with either
C. parvum
or
C. bovis
. Sequencing analysis of the gp60 gene showed all 67
C. parvum
samples belonged to subtype IIdA19G1. These findings suggested that the transmission of
Cryptosporidium
spp. in preweaned calves in Henan, China, appeared to be different from other areas both at genotype and subtype levels. Further molecular epidemiologic studies (including samples from both calves and humans) are needed to elucidate the transmission dynamics and public significance of
C. parvum
in cattle in China.
Efficient Differentiation of Mycobacterium abscessus Complex Isolates to the Species Level by a Novel PCR-Based Variable-Number Tandem-Repeat Assay
Authors: Choi Go-Eun , Chang Chulhun L. , Whang Jake , Kim Hwa-Jung , Kwon O. Jung , Koh Won-Jung , Shin Sung Jae
Published: December 2010

A novel duplex PCR method based on variable-number tandem-repeat targets to discriminate among
Mycobacterium abscessus
complex isolates was developed and evaluated in 85 clinical isolates. The assay accuracy was confirmed by a multiple-target sequence analysis. The duplex PCR assay is a one-step, reliable, and accurate assay for discriminating
M. abscessus
species.
Role of the Clinical Mycobacteriology Laboratory in Diagnosis and Management of Tuberculosis in Low-Prevalence Settings
Authors: Parrish Nicole M. , Carroll Karen C.
Published: December 2010

Tuberculosis (TB) remains a global epidemic, despite a significant decline in reported cases in the United States between 2008 and 2009. While the exact nature of this decline is unclear, one thing remains certain: TB, including multidrug-resistant (MDR) and extensively drug-resistant (XDR) TB, is no longer restricted to developing regions of the globe. It is of vital importance that both public and private mycobacteriology laboratories maintain the ability to detect and identify
Mycobacterium tuberculosis
from patient specimens, as well as correctly determine the presence of antibiotic resistance. To do this effectively requires careful attention to preanalytical, analytical, and postanalytical aspects of testing. Respiratory specimens require digestion and concentration followed by fluorescence microscopy. The Centers for Disease Control and Prevention (CDC) recommends the performance of a direct nucleic acid amplification method, regardless of smear results, on specimens from patients in whom the suspicion of tuberculosis is high. Liquid-based technologies are more rapid and sensitive for the detection of
M. tuberculosis
in culture and nucleic acid probes, but biochemicals are preferred for identification once growth is detected. Susceptibility testing is most often done using either the agar proportion method or a commercial broth system. New genotypic and phenotypic methods of susceptibility testing include first- and second-line agents and are promising, though not yet widely available. Finally, gamma interferon release assays are preferred to the tuberculin skin test for screening certain at-risk populations, and new CDC guidelines are available that assist clinicians in their use.
Diagnostic Performance of Filter Paper Lesion Impression PCR for Secondarily Infected Ulcers and Nonulcerative Lesions Caused by Cutaneous Leishmaniasis
Authors: Boggild Andrea K. , Ramos Ana Pilar , Valencia Braulio Mark , Veland Nicolas , Calderon Flor , Arevalo Jorge , Low Donald E. , Llanos-Cuentas Alejandro
Published: December 2010

We compared traditional cutaneous leishmaniasis diagnostic methods to filter paper lesion impression (FPLI) PCR for secondarily infected ulcers and nonulcerative lesions. The sensitivity and specificity of FPLI PCR for secondarily infected lesions (
n
= 8) were 100%. In primarily nonulcerative lesions (
n
= 15), the sensitivity of FPLI PCR was inferior to that of pooled-invasive-specimen PCR (72.7% versus 100%) (
P
= 0.10). FPLI PCR is sensitive, specific, and unlike invasive procedures, can be used in secondarily infected ulcers. Invasive specimen collection is superior in nonulcerative lesions.
Diversity of Interferon Antagonist Activities Mediated by NSP1 Proteins of Different Rotavirus Strains
Authors: Arnold Michelle M. , Patton John T.
Published: December 2010

Studies involving limited numbers of rotavirus (RV) strains have shown that the viral gene 5 product, NSP1, can antagonize beta interferon (IFN-β) expression by inducing the degradation of IFN-regulatory factors (IRFs) (IRF3, IRF5, and IRF7) or a component of the E3 ubiquitin ligase complex responsible for activating NF-κB (β-transducin repeat-containing protein [β-TrCP]). To gain a broader perspective of NSP1 activities, we examined various RV strains for the ability to inhibit IFN-β expression in human cells. We found that all strains encoding wild-type NSP1 impeded IFN-β expression but not always through IRF3 degradation. To identify other degradation targets involved in suppressing IFN-β expression, we used transient expression vectors to test the abilities of a diverse collection of NSP1 proteins to target IRF3, IRF5, IRF7, and β-TrCP for degradation. The results indicated that human RVs rely predominantly on the NSP1-induced degradation of IRF5 and IRF7 to suppress IFN signaling, whereas NSP1 proteins of animal RVs tended to target IRF3, IRF5, and IRF7, allowing the animal viruses a broader attack on the IFN-β signaling pathway. The results also suggested that the NSP1-induced degradation of β-TrCP is an uncommon mechanism of subverting IFN-β signaling but is one that can be shared with NSP1 proteins that induce IRF degradation. Our analysis reveals that the activities of NSP1 proteins are diverse, with no obvious correlations between degradations of pairs of target proteins. Thus, RVs have evolved functionally distinct approaches for subverting the host antiviral response, a property consistent with the immense sequence variation noted for NSP1 proteins.
Tupaia CD81, SR-BI, Claudin-1, and Occludin Support Hepatitis C Virus Infection
Authors: Tong Yimin , Zhu Yongzhe , Xia Xueshan , Liu Yuan , Feng Yue , Hua Xian , Chen Zhihui , Ding Hui , Gao Li , Wang Yongzhi , Feitelson Mark A. , Zhao Ping , Qi Zhong-Tian
Published: December 2010

Hepatitis C virus (HCV)-related research has been hampered by the lack of appropriate small-animal models. It has been reported that tree shrews, or tupaias (Tupaia belangeri), can be infected with serum-derived HCV. However, these reports do not firmly establish the tupaia as a reliable model of HCV infection. Human CD81, scavenger receptor class B type I (SR-BI), claudin 1 (CLDN1), and occludin (OCLN) are considered essential receptors or coreceptors for HCV cell entry. In the present study, the roles of these tupaia orthologs in HCV infection were assessed. Both CD81 and SR-BI of tupaia were found to be able to bind with HCV envelope protein 2 (E2). In comparison with human CD81, tupaia CD81 exhibited stronger binding activity with E2 and increased HCV pseudoparticle (HCVpp) cell entry 2-fold. The 293T cells transfected with tupaia CLDN1 became susceptible to HCVpp infection. Moreover, simultaneous transfection of the four tupaia factors into mouse NIH 3T3 cells made the cells susceptible to HCVpp infection. HCVpp of diverse genotypes were able to infect primary tupaia hepatocytes (PTHs), and this infection could be blocked by either anti-CD81 or anti-SR-BI. PTHs could be infected by cell culture-produced HCV (HCVcc) and did produce infectious progeny virus in culture supernatant. These findings indicate that PTHs possess all of the essential factors required for HCV entry and support the complete HCV infection cycle. This highlights both the mechanisms of susceptibility of tupaia to HCV infection and the possibility of using tupaia as a promising small-animal model in HCV study.
Transcriptional Profiling of CD4 T Cells Identifies Distinct Subgroups of HIV-1 Elite Controllers
Authors: Vigneault Francois , Woods Matthew , Buzon Maria J. , Li Chun , Pereyra Florencia , Crosby Seth D. , Rychert Jennifer , Church George , Martinez-Picado Javier , Rosenberg Eric S. , Telenti Amalio , Yu Xu G. , Lichterfeld Mathias
Published: December 2010

Human immunodeficiency virus type 1 (HIV-1) elite controllers maintain undetectable levels of viral replication in the absence of antiretroviral therapy (ART), but their underlying immunological and virological characteristics may vary. Here, we used a whole-genome transcriptional profiling approach to characterize gene expression signatures of CD4 T cells from an unselected cohort of elite controllers. The transcriptional profiles for the majority of elite controllers were similar to those of ART-treated patients but different from those of HIV-1-negative persons. Yet, a smaller proportion of elite controllers showed an alternative gene expression pattern that was indistinguishable from that of HIV-1-negative persons but different from that of highly active antiretroviral therapy (HAART)-treated individuals. Elite controllers with the latter gene expression signature had significantly higher CD4 T cell counts and lower levels of HIV-1-specific CD8
+
T cell responses but did not significantly differ from other elite controllers in terms of HLA class I alleles, HIV-1 viral loads determined by ultrasensitive single-copy PCR assays, or chemokine receptor polymorphisms. Thus, these data identify a specific subgroup of elite controllers whose immunological and gene expression characteristics approximate those of HIV-1-negative persons.
Complementing Defective Viruses That Express Separate Paramyxovirus Glycoproteins Provide a New Vaccine Vector Approach
Authors: Chattopadhyay Anasuya , Rose John K.
Published: December 2010

Replication-defective vaccine vectors based on vesicular stomatitis virus (VSV) lacking its envelope glycoprotein gene (G) are highly effective in animal models. However, such ΔG vectors are difficult to grow because they require complementation with the VSV G protein. In addition, the complementing G protein induces neutralizing antibodies in animals and thus limits multiple vector applications. In the process of generating an experimental Nipah virus (a paramyxovirus) vaccine, we generated two defective VSVΔG vectors, each expressing one of the two Nipah virus (NiV) glycoproteins (G and F) that are both required for virus entry to host cells. These replication-defective VSV vectors were effective at generating NiV neutralizing antibody in mice. Most interestingly, we found that these two defective viruses could be grown together and passaged in tissue culture cells in the absence of VSV G complementation. This mixture of complementing defective viruses was also highly effective at generating NiV neutralizing antibody in animals. This novel approach to growing and producing a vaccine from two defective viruses could be generally applicable to vaccine production for other paramyxoviruses or for other viruses where the expression of at least two different proteins is required for viral entry. Such an approach minimizes biosafety concerns that could apply to single, replication-competent VSV recombinants expressing all proteins required for infection.
Varicella-Zoster Virus Transcriptome in Latently Infected Human Ganglia
Authors: Nagel Maria A. , Choe Alexander , Traktinskiy Igor , Cordery-Cotter Robert , Gilden Don , Cohrs Randall J.
Published: December 2010

We recently developed a novel multiplex reverse transcription (RT)-PCR assay that allows rapid and sensitive detection of transcripts corresponding to all 68 unique varicella-zoster virus (VZV) open reading frames (ORFs) in only five amplification reactions (M. A. Nagel, D. Gilden, T. Shade, B. Gao, and R. J. Cohrs, J. Virol. Methods
157:
62-68, 2009). Herein, we applied multiplex RT-PCR analysis to mRNA extracted from 26 trigeminal ganglia latently infected with VZV and one control trigeminal ganglion negative for VZV DNA that were removed from 14 men and women, 16 to 84 years of age, within 24 h after death. Analysis identified VZV transcripts mapping to VZV ORFs 29, 62, and 63, previously detected and sequence verified; VZV ORFs 4 and 40, previously detected by
in situ
hybridization; and VZV ORFs 11, 41, 43, 57, and 68, not previously detected. VZV ORF 63 transcripts were the most prevalent. Comparison of the 10 VZV ORFs transcribed during latency to their herpes simplex virus type 1 homologues reveals that the latently transcribed VZV genes encode immediate-early, early, and late transcripts.
Structural Flexibility Allows the Functional Diversity of Potyvirus Genome-Linked Protein VPg
Authors: Rantalainen Kimmo I. , Eskelin Katri , Tompa Peter , Mäkinen Kristiina
Published: December 2010

Several viral genome-linked proteins (VPgs) of plant viruses are intrinsically disordered and undergo folding transitions in the presence of partners. This property has been postulated to be one of the factors that enable the functional diversity of the protein. We created a homology model ofPotato virus AVPg and positioned the known functions and structural properties of potyviral VPgs on the novel structural model. The model suggests an elongated structure with a hydrophobic core composed of antiparallel β-sheets surrounded by helices and a positively charged contact surface where most of the known activities are localized. The model most probably represents the fold induced immediately after binding of VPg to a negatively charged lipid surface or to SDS. When the charge of the positive surface was lowered by lysine mutations, the efficiencies ofin vitroNTP binding, uridylylation reaction, and unspecific RNA binding were reduced andin vivothe infectivity was debilitated. The most likely uridylylation site, Tyr63, locates to the positively charged surface. Surprisingly, a Tyr63Ala mutation did not prevent replication completely but blocked spreading of the virus. Based on the localization of Tyr119 in the model, it was hypothesized to serve as an alternative uridylylation site. Evidence to support the role of Tyr119 in replication was obtained which gives a positive example of the prediction power of the model. Taken together, our experimental data support the features presented in the model and the idea that the functional diversity is attributable to structural flexibility.
Orf Virus
ORFV121
Encodes a Novel Inhibitor of NF-κB That Contributes to Virus Virulence
Authors: Diel D. G. , Luo S. , Delhon G. , Peng Y. , Flores E. F. , Rock D. L.
Published: December 2010

Orf virus (ORFV), the type member of the genus
Parapoxvirus
of the
Poxviridae
, has evolved novel strategies (proteins and/or mechanisms of action) to modulate host cell responses regulated by the nuclear factor-κB (NF-κB) signaling pathway. Here, we present data indicating that ORFV
ORFV121
, a gene unique to parapoxviruses, encodes a novel viral NF-κB inhibitor that binds to and inhibits the phosphorylation and nuclear translocation of NF-κB-p65. The infection of cells with an
ORFV121
deletion mutant virus (OV-IA82Δ121) resulted in increased NF-κB-mediated gene transcription, and the expression of ORFV121 in cell cultures significantly suppressed NF-κB-regulated reporter gene expression. ORFV ORFV121 physically interacts with NF-κB-p65 in the cell cytoplasm, thus providing a mechanism for the inhibition of NF-κB-p65 phosphorylation and nuclear translocation. Notably, the deletion of
ORFV121
from the viral genome markedly decreased ORFV virulence and disease pathogenesis in sheep, indicating that ORFV121 is a virulence determinant for ORFV in the natural host.
EBNA3C Attenuates the Function of p53 through Interaction with Inhibitor of Growth Family Proteins 4 and 5
Authors: Saha Abhik , Bamidele Adebowale , Murakami Masanao , Robertson Erle S.
Published: December 2010

Epstein-Barr virus (EBV)-encoded EBNA3C is one of the latent proteins essential for the efficient transformation of human primary B lymphocytes into continuously proliferating lymphoblastoid cell lines (LCLs)
in vitro
through manipulation of a number of major cellular pathways. Although it does not have direct DNA-binding activity, EBNA3C plays a central role in the transcriptional modulation of a wide range of both viral and cellular genes during latent infection. Recently, we showed that EBNA3C can directly bind to the tumor suppressor protein p53 and repress its functions, in part by blocking its transcriptional activity as well as facilitating its degradation through stabilization of its negative regulator, Mdm2. In this study, we further showed that EBNA3C can negatively regulate p53-mediated functions by interacting with its regulatory proteins, the inhibitor of growth family proteins ING4 and ING5, shown to be frequently deregulated in different cancers. Functional mapping revealed that both ING4 and ING5 bound to N-terminal domain residues 129 to 200 of EBNA3C, which was previously demonstrated to associate with p53 and is also essential for LCL growth. In addition, we showed that a conserved domain of either ING4 or ING5 bound to both p53 and EBNA3C in a competitive manner, suggesting a potential role for EBNA3C whereby the ING4 or -5/p53 pathway is modulated in EBV-infected cells. Subsequently, we demonstrated that EBNA3C significantly suppresses both the ING4- and ING5-mediated regulation of p53 transcriptional activity in a dose-dependent manner. A colony formation assay as well as an apoptosis assay showed that EBNA3C nullified the negative regulatory effects on cell proliferation induced by coupled expression of p53 in the presence of either ING4 or ING5 in Saos-2 (p53
−/−
) cells. This report demonstrates a possible role for the candidate tumor suppressor ING genes in the biology of EBV-associated cancers.
VEGFR2 and Src Kinase Inhibitors Suppress Andes Virus-Induced Endothelial Cell Permeability
Authors: Gorbunova Elena E. , Gavrilovskaya Irina N. , Pepini Timothy , Mackow Erich R.
Published: December 2010

Hantaviruses predominantly infect human endothelial cells and, in the absence of cell lysis, cause two diseases resulting from increased vascular permeability. Andes virus (ANDV) causes a highly lethal acute pulmonary edema termed hantavirus pulmonary syndrome (HPS). ANDV infection enhances the permeability of endothelial cells in response to vascular endothelial growth factor (VEGF) by increasing signaling responses directed by the VEGFR2-Src-VE-cadherin pathway, which directs adherens junction (AJ) disassembly. Here we demonstrate that inhibiting pathway-specific VEGFR2 and Src family kinases (SFKs) blocks ANDV-induced endothelial cell permeability. Small interfering RNA (siRNA) knockdown of Src within ANDV-infected endothelial cells resulted in an ∼70% decrease in endothelial cell permeability compared to that for siRNA controls. This finding suggested that existing FDA-approved small-molecule kinase inhibitors might similarly block ANDV-induced permeability. The VEGFR2 kinase inhibitor pazopanib as well as SFK inhibitors dasatinib, PP1, bosutinib, and Src inhibitor 1 dramatically inhibited ANDV-induced endothelial cell permeability. Consistent with their kinase-inhibitory concentrations, dasatinib, PP1, and pazopanib inhibited ANDV-induced permeability at 1, 10, and 100 nanomolar 50% inhibitory concentrations (IC50s), respectively. We further demonstrated that dasatinib and pazopanib blocked VE-cadherin dissociation from the AJs of ANDV-infected endothelial cells by >90%. These findings indicate that VEGFR2 and Src kinases are potential targets for therapeutically reducing ANDV-induced endothelial cell permeability and, as a result, capillary permeability during HPS. Since the functions of VEGFR2 and SFK inhibitors are already well defined and FDA approved for clinical use, these findings rationalize their therapeutic evaluation for efficacy in reducing HPS disease. Endothelial cell barrier functions are disrupted by a number of viruses that cause hemorrhagic, edematous, or neurologic disease, and as a result, our findings suggest that VEGFR2 and SFK inhibitors should be considered for regulating endothelial cell barrier functions altered by additional viral pathogens.
Smoking cessation strategies in Crohn's disease
Authors: Wilson Andrew
Published: December 2010

Although smoking cessation is one of the most effective interventions in patients with Crohn's disease, the prevalence of smoking in these patients is higher than in the general population, and much higher than in patients with other smoking-related diseases. There is a lack of awareness of the benefits of cessation among patients and general practitioners, and a lack of engagement in smoking cessation by specialists. This article discusses the strategies that practitioners can use to motivate patients to quit and how the success of a quitting attempt can be maximised.
A comparative study of quantitative immunohistochemistry and quantum dot immunohistochemistry for mutation carrier identification in Lynch syndrome
Authors: Barrow Emma , Evans D Gareth , McMahon Ray , Hill James , Byers Richard
Published: December 2010

Aims

Lynch Syndrome is caused by mutations in DNA mismatch repair (MMR) genes. Mutation carrier identification is facilitated by immunohistochemical detection of the MMR proteins MHL1 and MSH2 in tumour tissue and is desirable as colonoscopic screening reduces mortality. However, protein detection by conventional immunohistochemistry (IHC) is subjective, and quantitative techniques are required. Quantum dots (QDs) are novel fluorescent labels that enable quantitative multiplex staining. This study compared their use with quantitative 3,3′-diaminobenzidine (DAB) IHC for the diagnosis of Lynch Syndrome.

Methods

Tumour sections from 36 mutation carriers and six controls were obtained. These were stained with DAB on an automated platform using antibodies against MLH1 and MSH2. Multiplex QD immunofluorescent staining of the sections was performed using antibodies against MLH1, MSH2 and smooth muscle actin (SMA). Multispectral analysis of the slides was performed. The staining intensity of DAB and QDs was measured in multiple colonic crypts, and the mean intensity scores calculated. Receiver operating characteristic (ROC) curves of staining performance for the identification of mutation carriers were evaluated.

Results

For quantitative DAB IHC, the area under the MLH1 ROC curve was 0.872 (95% CI 0.763 to 0.981), and the area under the MSH2 ROC curve was 0.832 (95% CI 0.704 to 0.960). For quantitative QD IHC, the area under the MLH1 ROC curve was 0.812 (95% CI 0.681 to 0.943), and the area under the MSH2 ROC curve was 0.598 (95% CI 0.418 to 0.777).

Conclusions

Despite the advantage of QD staining to enable several markers to be measured simultaneously, it is of lower utility than DAB IHC for the identification of MMR mutation carriers. Automated DAB IHC staining and quantitative slide analysis may enable high-throughput IHC.
Carboxylesterase 1 (Ces1): from monocyte marker to major player
Authors: Markey Geraldine M
Published: December 2010

There are few, if any, enzymes that have been studied by, and have importance in, so many and varied disciplines as has monocyte esterase/Ces 1. In this review its involvement in the fields of histochemistry, haematology, toxicology, pharmacology, therapeutics, and tumour cell killing, immune surveillance and malignant disease, is briefly described.
Donor cell origin of multiple myeloma occurring after allogeneic haematopoietic stem cell transplantation in a patient with refractory anaemia with ring sideroblast
Authors: Kim Young-Il , Kim Hye-Ran , Shin Myung-Geun , Lee Young-Jin , Shin Jong-Hee , Suh Soon-Pal , Ryang Dong-Wook
Published: December 2010

Post-transplant lymphoproliferative disorder (PTLD) is a rare but life-threatening complication after solid organ and haematopoietic stem cell transplantation. A 40-year-old woman who was diagnosed as having refractory anaemia with ring sideroblast 6 years ago took an ABO mismatched, unrelated allogeneic haematopoietic stem cell transplantation (HSCT) from a 32-year-old healthy male donor. The bone marrow (BM) study was carried out because of progressing pancytopenia, serum biclonal gammopathy and a distorted ratio of serum level of free κ and λ light chain 138 days after HSCT. The BM examination showed an increased number of plasma cells (12% of total marrow cells) comprising mainly CD45−CD19−CD138+ malignant plasma cells with an immunoglobulin heavy-chain gene rearrangement. Conventional cytogenetics and molecular personal identification studies revealed that all BM cells were totally replaced by donor cells, thus indicating the donor cell origin of PTLD-multiple myeloma. The BM microenvironment of the recipient might be associated with the development of PTLD-multiple myeloma.
Relationship between pathological features, HER2 protein expression andHER2and CEP17 copy number in breast cancer: biological and methodological considerations
Authors: Lambein Kathleen , Praet Marleen , Forsyth Ramses , Van den Broecke Rudy , Braems Geert , Matthys Bart , Cocquyt Veronique , Denys Hannelore , Pauwels Patrick , Libbrecht Louis
Published: December 2010

Aims

A few reports have assessed HER2 status in breast cancer by both dual-probe fluorescence in situ hybridisation (FISH) and immunohistochemistry (IHC) in an unselected and consecutive fashion, but CEP17 andHER2copy number were not evaluated separately in these studies. Therefore, the aim of this study was to perform FISH testing forHER2in a large number of breast tumours, irrespective of the IHC scores, which were also determined in all cases.

Methods

Both FISH and IHC were applied to 200 tumours from 196 consecutive patients who underwent resection of primary breast cancer with the sentinel procedure and/or axillary dissection. Not only the ratio, but also meanHER2and CEP17 copy number were determined and used in statistical analyses to evaluate relationships between FISH, IHC and clinicopathological features.

Results

The amplification status based solely on HER2 signals was 98% concordant with results of dual-probe FISH. In non-amplified tumours, the mean CEP17 andHER2copy number correlated, possibly because of cell cycling. Amplified tumours were histopathologically more aggressive than non-amplified tumours, and features of aggressiveness increased with the meanHER2copy number. In both amplified and non-amplified tumours, a gene dosage effect was observed: an increase in the meanHER2copy number was associated with a higher IHC score.

Conclusions

This working method and analysis enabled new insights to be obtained into the pathobiology of HER2 in breast cancer. The findings may be helpful in optimising the methodology of HER2 testing.
A literary portrait of a case of Hodgkin lymphoma by Wilkie Collins?
Authors: Fernandez-Flores Angel
Published: December 2010

In October 1859, ‘The Queen of Hearts’ was published by Wilkie Collins. This book is a collection of 10 short stories written by the protagonists (three retired brothers), in order to keep their niece (aka the queen of hearts) entertained, until the arrival of her fiancé-to-be. The second story of the book, titled ‘Brother Griffith's story of the family secret,’ narrates the death of a young girl who has a slow-growing deforming tumour on her neck. Even with the few clinical details provided by the literary description, the current report examines the possibility of Hodgkin lymphoma as a diagnosis, concluding that the option is not an unwise one.
Digital mammography in a screening programme and its implications for pathology: a comparative study
Authors: Feeley Linda , Kiernan Donal , Mooney Therese , Flanagan Fidelma , Hargaden Gormlaith , Kell Malcolm , Stokes Maurice , Kennedy Margaret
Published: December 2010

Aims

Most studies comparing full-field digital mammography (FFDM) with conventional screen-film mammography (SFM) have been radiology-based. The pathological implications of FFDM have received little attention in the literature, especially in the context of screening programmes. The primary objective of this retrospective study is to compare FFDM with SFM in a population-based screening programme with regard to a number of pathological parameters.

Methods

During the study period, 107 818 women underwent screening mammograms with almost equal numbers obtained with each technique (49.9% with SFM vs 50.1% with FFDM). We compared SFM with FFDM using the following parameters: recall rate, diagnostic core biopsy rate, cancer detection rates, B3 rate, B4 rate, preoperative diagnostic rate for malignancy, positive predictive values and tumour characteristics.

Results

The recall rate was significantly higher with FFDM (4.21% vs 3.52%, p<0.0001). The overall cancer detection rate of 7.2 per 1000 women screened with FFDM was also significantly higher than the rate of 6.2 per 1000 women screened with SFM (p=0.04). The B3 rate in the SFM group was 1.3 per 1000 women screened versus 2.5 per 1000 women screened in the FFDM group (p<0.001). The recall rate and cancer detection rates (overall, invasive and pure ductal carcinoma in situ) were all significantly higher with FFDM for lesions presenting as microcalcifications.

Conclusions

The higher cancer detection rate with FFDM in this study was due to improved detection of microcalcifications. However, this was achieved at the cost of a higher recall rate and a higher B3 rate, indicating that overtreatment may be problematic with digital mammography.
Participatory ergonomics to reduce exposure to psychosocial and physical risk factors for low back pain and neck pain: results of a cluster randomised controlled trial
Authors: Driessen Maurice T , Proper Karin I , Anema Johannes R , Knol Dirk L , Bongers Paulien M , van der Beek Allard J
Published: December 2010

Objectives

This study investigated the effectiveness of the Stay@Work participatory ergonomics programme to reduce workers′ exposure to psychosocial and physical risk factors.

Methods

37 departments (n=3047 workers) from four Dutch companies participated in this cluster randomised controlled trial; 19 (n=1472 workers) were randomised to an intervention group (participatory ergonomics) and 18 (n=1575 workers) to a control group (no participatory ergonomics). During a 6 h meeting guided by an ergonomist, working groups devised ergonomic measures to reduce psychosocial and physical workload and implemented them within 3 months in their departments. Data on psychosocial and physical risk factors for low back pain and neck pain were collected at baseline and after 6 months. Psychosocial risk factors were measured using the Job Content Questionnaire and physical risk factors using the Dutch Musculoskeletal Questionnaire. Intervention effects were studied using multilevel analysis.

Results

Intervention group workers significantly increased on decision latitude (0.29 points; 95% CI 0.07 to 0.52) and decision authority (0.16 points; 95% CI 0.04 to 0.28) compared to control workers. However, exposure to awkward trunk working postures significantly increased in the intervention group (OR 1.86; 95% CI 1.15 to 3.01) compared to the control group. No significant differences between the intervention and control group were found for the remaining risk factors. After 6 months, loss to follow-up was 35% in the intervention group and 29% in the control group.

Conclusion

Participatory ergonomics was not effective in reducing exposure to psychosocial and physical risk factors for low back pain and neck pain among a large group of workers.

Trial registration

ISRCTN27472278.
Not just ‘a few wisps’: real-time measurement of tobacco smoke at entrances to office buildings
Authors: Kaufman Pamela , Zhang Bo , Bondy Susan J , Klepeis Neil , Ferrence Roberta
Published: December 2010

Introduction

An unintended consequence of indoor smoking restrictions is the relocation of smoking to building entrances, where non-smokers may be exposed to secondhand smoke, and smoke from outdoor areas may drift through entrances, exposing people inside. Tobacco smoke has been linked to numerous health effects in non-smokers and there is no safe level of secondhand smoke (SHS) exposure. This paper presents data on levels of tobacco smoke inside and outside entrances to office buildings.

Methods

Real-time air quality monitors were used to simultaneously measure respirable particulate matter (PM2.5; air pollutant particles with a diameter of 2.5 μm or less) as a marker for tobacco smoke, outside and inside 28 entrances to office buildings in downtown Toronto, Ontario, in May and June 2008. Measurements were taken when smoking was and was not present within 9 m of entrances. Background levels of PM2.5were also measured for each session. A mixed model analysis was used to estimate levels of PM2.5, taking into account repeated measurement errors.

Results

Peak levels (10 s averages) of PM2.5were as high as 496 μg/m3when smoking was present. Mixed model analysis shows that the average outdoor PM2.5with smoking was significantly higher than the background level (p<0.0001), and significantly and positively associated with the number of lit cigarettes (p<0.0001). The average level of PM2.5with ≥5 lit cigarettes was 2.5 times greater than the average background level.

Conclusions

These findings support smoke-free policies at entrances to buildings to protect non-smokers from exposure to tobacco smoke.
Tailored reference limits for urine corticoid:creatinine ratio in dogs to answer distinct clinical questions
Authors: Zeugswetter F. , Bydzovsky N. , Kampner D. , Schwendenwein I.
Published: December 2010

To establish reference intervals for the urinary corticoid:creatinine ratio (UCCR) determined by chemiluminometric immunoassay, UCCR was measured by this method in 50 healthy dogs. To assess the diagnostic performance of different cut‐off levels, the UCCR of 66 dogs with hyperadrenocorticism and 87 dogs with diseases mimicking hyperadrenocorticism were used to construct a receiver operating characteristic (ROC) curve. The upper reference limit derived from morning samples in healthy dogs was 30.81 × 10−6. The area under the ROC curve was 0.94. The diagnostic cut‐off with the highest negative likelihood ratio was 26.5 × 10−6(sensitivity 1, specificity 0.54), whereas the cut‐off with the highest positive likelihood ratio was 161.2 × 10−6(specificity 0.988, sensitivity 0.515). The application of these two different diagnostic cut‐offs eliminated the necessity to perform additional tests in 53 per cent of the patient population.
Retrospective detection by negative contrast electron microscopy of faecal viral particles in free‐living wild red squirrels (Sciurus vulgaris) with suspected enteropathy in Great Britain
Authors: Everest D. J. , Stidworthy M. F. , Milne E. M. , Meredith A. L. , Chantrey J. , Shuttleworth C. , Blackett T. , Butler H. , Wilkinson M. , Sainsbury A. W.
Published: December 2010

Transmission electron microscopy identified adenovirus particles in 10 of 70 (14.3 per cent) samples of large intestinal content collected at postmortem examination from free‐living wild red squirrels (Sciurus vulgaris) across Great Britain between 2000 and 2009. Examination was limited to cases in which an enteropathy was suspected on the basis of predetermined macroscopic criteria such as semi‐solid or diarrhoeic faeces, suspected enteritis or the presence of intussusception. In most cases, meaningful histological examination of enteric tissue was not possible due to pronounced autolysis. Two (2.9 per cent) of the samples were negative for adenovirus but were found to contain rotavirus particles, a novel finding in this species.