bluuebunny/crossref_metadata_2025_split huggingface dataset:
Epidemiology and Clinical Presentations of the Four Human Coronaviruses 229E, HKU1, NL63, and OC43 Detected over 3 Years Using a Novel Multiplex Real-Time PCR Method
Authors: Gaunt E. R. , Hardie A. , Claas E. C. J. , Simmonds P. , Templeton K. E. 
Published: June 2010

ABSTRACT
Four human coronaviruses (HCoV-229E, HCoV-HKU1, HCoV-NL63, and HCoV-OC43) are associated with a range of respiratory outcomes, including bronchiolitis and pneumonia. Their epidemiologies and clinical characteristics are poorly described and are often reliant on case reports. To address these problems, we conducted a large-scale comprehensive screening for all four coronaviruses by analysis of 11,661 diagnostic respiratory samples collected in Edinburgh, United Kingdom, over 3 years between July 2006 and June 2009 using a novel four-way multiplex real-time reverse transcription-PCR (RT-PCR) assay. Coronaviruses were detected in 0.3 to 0.85% of samples in all age groups. Generally, coronaviruses displayed marked winter seasonality between the months of December and April and were not detected in summer months, which is comparable to the pattern seen with influenza viruses. HCoV-229E was the exception; detection was confined to the winter of 2008 and was sporadic in the following year. There were additional longer-term differences in detection frequencies between seasons, with HCoV-OC43 predominant in the first and third seasons and HCoV-HKU1 dominating in the second (see Results for definitions of seasons). A total of 11 to 41% of coronaviruses detected were in samples testing positive for other respiratory viruses, although clinical presentations of coronavirus monoinfections were comparable to those of viruses which have an established role in respiratory disease, such as respiratory syncytial virus, influenza virus, and parainfluenza viruses. The novel multiplex assay for real-time pan-coronavirus detection enhances respiratory virus diagnosis, overcomes potential diagnostic problems arising through seasonal variation in coronavirus frequency, and provides novel insights into the epidemiology and clinical implications of coronaviruses.
Detection and Characterization of Human
Ureaplasma
Species and Serovars by Real-Time PCR
Authors: Xiao Li , Glass John I. , Paralanov Vanya , Yooseph Shibu , Cassell Gail H. , Duffy Lynn B. , Waites Ken B. 
Published: June 2010

ABSTRACT

We designed primers and probes for the detection and discrimination of
Ureaplasma parvum
and
U. urealyticum
and their 14 serovars by real-time PCR. The analytical sensitivity and specificity of the multiplex species-specific PCR were determined by testing corresponding American Type Culture Collection (ATCC) type strains, 47 other microbial species, and human genomic DNA. The limits of the multiplex PCR were 2.8 × 10
−2
CFU/μl PCR mixture for detecting
U. parvum
and 4.1 × 10
−2
CFU/μl PCR mixture for detecting
U. urealyticum
. Clinical specificity and sensitivity were proven by comparison with culture and traditional PCR. For the detection of any
Ureaplasma
species, the clinical sensitivity and specificity of real-time PCR were 96.9% and 79.0%, respectively, using culture as a reference. Multiplex real-time PCR was also more sensitive than traditional PCR in discriminating the two
Ureaplasma
species in culture-positive subcultures. Each of the 14 monoplex serovar-specific PCR assays was specific for the corresponding ATCC type strain serovar. This new species identification PCR is specific and sensitive in the detection of
Ureaplasma
species in clinical specimens, and the serovar-specific PCR assays are the first set of complete genotypic assays to differentiate all 14 known
Ureaplasma
serovars. These assays provide quick and reliable means for investigating the epidemiology and pathogenicity of ureaplasmas at the serovar level.
Comparison of the Novel Partec Rapid Malaria Test to the Conventional Giemsa Stain and the Gold Standard Real-Time PCR
Authors: Nkrumah Bernard , Agyekum Alex , Acquah Samuel E. K. , May Jürgen , Tannich Egbert , Brattig Norbert , Nguah Samuel Blay , von Thien Heidrun , Adu-Sarkodie Yaw , Huenger Frank 
Published: June 2010

ABSTRACT

Malaria remains the single most frequent cause of death in Africa, killing one child every 30 s, but treatment decisions are often made only on clinical diagnosis, as laboratory techniques to confirm the clinical suspicion are labor intensive and costly. In this study, we evaluated the recently developed Partec rapid malaria test (PM) for the detection of
Plasmodium
spp. in human blood from patients in an area where malaria is endemic and compared the results with those of thick blood film Giemsa stain (GS) in terms of its performance and operational characteristics, using real-time (RT) PCR as the gold standard. The sensitivities of the PM and the GS were 62.2% (95% CI, 56.3 to 67.8) and 61.8% (95% CI, 55.9 to 67.4), respectively, while the specificities were 96.0% (95% CI, 92.3 to 98.3) and 98% (95% CI, 95.0 to 99.5), respectively. There was an excellent agreement between the results for the PM and those of the GS (
k
[level of agreement] = 0.96;
P
< 0.001). The results for the PM were obtained more quickly and at less cost than those for the GS. The performance characteristics of the PM were almost equal to those of the GS, but the operational characteristics were better, and the PM can therefore be considered as an alternative method for GS.
Tick-Borne Encephalitis Virus Delays Interferon Induction and Hides Its Double-Stranded RNA in Intracellular Membrane Vesicles
Authors: Överby Anna K. , Popov Vsevolod L. , Niedrig Matthias , Weber Friedemann 
Published: June 2010

ABSTRACT

Tick-borne encephalitis virus (TBEV) (family
Flaviviridae
, genus
Flavivirus
) accounts for approximately 10,000 annual cases of severe encephalitis in Europe and Asia. Here, we investigated the induction of the antiviral type I interferons (IFNs) (alpha/beta IFN [IFN-α/β]) by TBEV. Using strains Neudörfl, Hypr, and Absettarov, we demonstrate that levels of IFN-β transcripts and viral RNA are strictly correlated. Moreover, IFN induction by TBEV was dependent on the transcription factor IFN regulatory factor 3 (IRF-3). However, even strain Hypr, which displayed the strongest IFN-inducing activity and the highest RNA levels, substantially delayed the activation of IRF-3. As a consequence, TBEV can keep the level of IFN transcripts below the threshold value that would permit the release of IFN by the cell. Only after 24 h of infection have cells accumulated sufficient IFN transcripts to produce detectable amounts of secreted IFNs. The delay in IFN induction appears not to be caused by a specific viral protein, since the individual expressions of TBEV C, E, NS2A, NS2B, NS3, NS4A, NS4B, NS5, and NS2B-NS3, as well as TBEV infection itself, had no apparent influence on specific IFN-β induction. We noted, however, that viral double-stranded RNA (dsRNA), an important trigger of the IFN response, is immunodetectable only inside intracellular membrane compartments. Nonetheless, the dependency of IFN induction on IFN promoter stimulator 1 (IPS-1) as well as the phosphorylation of the alpha subunit of eukaryotic initiation factor 2 (eIF2α) suggest the cytoplasmic exposure of some viral dsRNA late in infection. Using ultrathin-section electron microscopy, we demonstrate that, similar to other flaviviruses, TBEV rearranges intracellular membranes. Virus particles and membrane-connected vesicles (which most likely represent sites of virus RNA synthesis) were observed inside the endoplasmic reticulum. Thus, apparently, TBEV rearranges internal cell membranes to provide a compartment for its dsRNA, which is largely inaccessible for detection by cytoplasmic pathogen receptors. This delays the onset of IFN induction sufficiently to give progeny particle production a head start of approximately 24 h.
Genetic Diversity and Histo-Blood Group Antigen Interactions of Rhesus Enteric Caliciviruses
Authors: Farkas Tibor , Cross Robert W. , Hargitt Edwin , Lerche Nicholas W. , Morrow Ardythe L. , Sestak Karol 
Published: June 2010

ABSTRACTRecently, we reported the discovery and characterization of Tulane virus (TV), a novel rhesus calicivirus (CV) (T. Farkas, K. Sestak, C. Wei, and X. Jiang, J. Virol.82:5408-5416, 2008). TV grows well in tissue culture, and it represents a new genus withinCaliciviridae, with the proposed name ofRecovirus. We also reported a high prevalence of CV antibodies in macaques of the Tulane National Primate Research Center (TNPRC) colony, including anti-norovirus (NoV), anti-sapovirus (SaV), and anti-TV (T. Farkas, J. Dufour, X. Jiang, and K. Sestak, J. Gen. Virol. 91:734-738, 2010). To broaden our knowledge about CV infections in captive nonhuman primates (NHP), 500 rhesus macaque stool samples collected from breeding colony TNPRC macaques were tested for CVs. Fifty-seven (11%) samples contained recovirus isolates. In addition, one NoV was detected. Phylogenetic analysis classified the recovirus isolates into two genogroups and at least four genetic types. The rhesus NoV isolate was closely related to GII human NoVs. TV-neutralizing antibodies were detected in 88% of serum samples obtained from primate caretakers. Binding and plaque reduction assays revealed the involvement of type A and B histo-blood group antigens (HBGA) in TV infection. Taken together, these findings indicate the zoonotic potential of primate CVs. The discovery of a genetically diverse and prevalent group of primate CVs and remarkable similarities between rhesus enteric CVs and human NoVs opens new possibilities for research involvingin vitroandin vivomodels of human NoV gastroenteritis.
Simian Immunodeficiency Virus Infection Induces Expansion of α4β7+and Cytotoxic CD56+NK Cells
Authors: Reeves R. Keith , Evans Tristan I. , Gillis Jacqueline , Johnson R. Paul 
Published: June 2010

ABSTRACTHerein we demonstrate that chronic simian immunodeficiency virus (SIV) infection induces significant upregulation of the gut-homing marker α4β7 on macaque NK cells, coupled with downregulation of the lymph node-trafficking marker, CCR7. Interestingly, in naïve animals, α4β7 expression was associated with increased NK cell activation and, on CD16+NK cells, delineated a unique dual-function cytotoxic-CD107a+/gamma interferon (IFN-γ)-secreting population. However, while SIV infection increased CD107a expression on stimulated CD56+NK cells, α4β7+and α4β7−NK cells were affected similarly. These findings suggest that SIV infection redirects NK cells away from the lymph nodes to the gut mucosae but alters NK cell function independent of trafficking repertoires.
Coxsackievirus B3 Infection Activates the Unfolded Protein Response and Induces Apoptosis through Downregulation of p58
IPK
and Activation of CHOP and SREBP1
Authors: Zhang Huifang M. , Ye Xin , Su Yue , Yuan Ji , Liu Zhen , Stein David A. , Yang Decheng 
Published: June 2010

ABSTRACT

Cardiomyocyte apoptosis is a hallmark of coxsackievirus B3 (CVB3)-induced myocarditis. We used cardiomyocytes and HeLa cells to explore the cellular response to CVB3 infection, with a focus on pathways leading to apoptosis. CVB3 infection triggered endoplasmic reticulum (ER) stress and differentially regulated the three arms of the unfolded protein response (UPR) initiated by the proximal ER stress sensors ATF6a (activating transcription factor 6a), IRE1-XBP1 (X box binding protein 1), and PERK (PKR-like ER protein kinase). Upon CVB3 infection, glucose-regulated protein 78 expression was upregulated, and in turn ATF6a and XBP1 were activated via protein cleavage and mRNA splicing, respectively. UPR activity was further confirmed by the enhanced expression of UPR target genes ERdj4 and EDEM1. Surprisingly, another UPR-associated gene, p58
IPK
, which often is upregulated during infections with other types of viruses, was downregulated at both mRNA and protein levels after CVB3 infection. These findings were observed similarly for uninfected Tet-On HeLa cells induced to overexpress ATF6a or XBP1. In exploring potential connections between the three UPR pathways, we found that the ATF6a-induced downregulation of p58
IPK
was associated with the activation of PKR (PERK) and the phosphorylation of eIF2α, suggesting that p58
IPK
, a negative regulator of PERK and PKR, mediates cross-talk between the ATF6a/IRE1-XBP1 and PERK arms. Finally, we found that CVB3 infection eventually produced the induction of the proapoptoic transcription factor CHOP and the activation of SREBP1 and caspase-12. Taken together, these data suggest that CVB3 infection activates UPR pathways and induces ER stress-mediated apoptosis through the suppression of P58
IPK
and induction/activation of CHOP, SREBP1, and caspase-12.
Effects of Anti-VEGF Treatment Duration on Tumor Growth, Tumor Regrowth, and Treatment Efficacy
Authors: Bagri Anil , Berry Leanne , Gunter Bert , Singh Mallika , Kasman Ian , Damico Lisa A. , Xiang Hong , Schmidt Maike , Fuh Germaine , Hollister Beth , Rosen Oliver , Plowman Greg D. 
Published: June 2010

Abstract
Purpose: Inhibition of the vascular endothelial growth factor (VEGF) axis is the basis of all currently approved antiangiogenic therapies. In preclinical models, anti-VEGF blocking antibodies have shown broad efficacy that is dependent on both tumor context and treatment duration. We aimed to characterize this activity and to evaluate the effects of discontinuation of treatment on the dynamics of tumor regrowth.
Experimental Design: We evaluated the effects of anti-VEGF treatment on tumor growth and survival in 30 xenograft models and in genetic mouse models of cancer. Histologic analysis was used to evaluate the effects of treatment on tumor vasculature. We used a variety of treatment regimens to allow analysis of the effects of treatment duration and cessation on growth rate, survival, and vascular density.
Results: Preclinical tumor models were characterized for their varied dependence on VEGF, thereby defining models for testing other agents that may complement or augment anti-VEGF therapy. We also found that longer exposure to anti-VEGF monoclonal antibodies delayed tumor growth and extended survival in established tumors from both cell transplants and genetic tumor models and prevented regrowth of a subset of residual tumors following cytoablative therapy. Discontinuation of anti-VEGF in established tumors resulted in regrowth at a rate slower than that in control-treated animals, with no evidence of accelerated tumor growth or rebound. However, more rapid regrowth was observed following discontinuation of certain chemotherapies. Concurrent administration of anti-VEGF seemed to normalize these accelerated growth rates.
Conclusions: In diverse preclinical models, continuous VEGF suppression provides maximal benefit as a single agent, combined with chemotherapy, or as maintenance therapy once chemotherapy has been stopped. Clin Cancer Res; 16(15); 3887–900. ©2010 AACR.
“By the Mouth of Two or Three Witnesses” Paul’s Invocation of a Deuteronomic Statute
Authors: Welborn Laurence L. 
Published: June 2010

AbstractThe history of interpretation of 2 Cor 13:1 has been dominated by a metaphorical construal of the “three witnesses” as Paul’s three visits to Corinth. This essay proposes a literal understanding of the text: Paul invokes the Deuteronomic rule of judicial witnesses in his own defense, in accordance with the purposes of the statute. The essay then explores the implications of this interpretation for reconstruction of what happened on the occasion of Paul’s second, “painful” visit to Corinth.
“Because for a long time (the Gods of Zalpa) have been ignored ..Hence these offerings in this way do we donate”. New Celebrations in the Zalpuwa Land
Authors: Corti Carlo 
Published: June 2010

AbstractIn this contribution a new group of documents, individualized by the writer, is introduced. It is connected to the ancient city of Zalpa on the Black Sea and is entitled “Celebrations in the Zalpuwa Land” (CTH 667): the religious ceremonies described, as well as several participants and the majority of the places mentioned in these tablets, don’t find parallel in Hittite documentation known to us. On the basis of the analyzed elements, it is proposed to assign the colophon of the funerary ritual for the sovereign, IBoT 2.130 to these texts; moreover, it is held that, unlike the scenario that up until now has been reconstructed, the region of Zalpa was recovered once again by the Hittites in the imperial period.
Blurring of the Public/Private Divide: The Canadian Chapter
Authors: Thomas Bryan , Flood Colleen M. 
Published: June 2010

AbstractBlurring of public/private divide is occurring in different ways around the world, with differential effects in terms of access and equity. In Canada, one pathway towards privatization has received particular attention: duplicative private insurance, allowing those with the financial means to bypass queues in the public system. We assess recent legal and policy developments on this front, but also describe other trends towards the blurring of public and private in Canada: the reliance on mandated private insurance for pharmaceutical coverage; provincial governments’ reliance on public-private partnerships to finance hospitals; and the incorporation of for-profit clinics within the public health care system.
Health System Reforms in The Netherlands: From Public to Private and its Effects on Equal Access to Health Care
Authors: den Exter Andre 
Published: June 2010

AbstractDutch health system reforms can be characterized by an ambiguous approach, combining competition and private initiatives on several health care markets with restrictive measures inspired by social values. Respecting equal access to necessary health care services on a competitive market appears to be an extremely difficult and bureaucratic exercise, if not impossible.
Holy Orders: Nehemiah Goreh's Ordination Ordeal and the Problem of 'Social Distance' in Nineteenth-Century North Indian Anglicanism
Authors: Young Richard Fox 
Published: June 2010

AbstractUsing the example of Nehemiah Goreh, a mid nineteenth-century Brahmin Hindu convert to Christianity, the essay explores how Anglican missionaries interacted with Indian counterparts, sometimes encouraging their ordination (as was the case in the South), or (as was the case in the North) placing obstacles in their way. After an agonistically 'cognitive' struggle with Christian faith, Goreh was recommended for ordination by the Low-Church Anglican missionaries of Benares, only to be denied 'Holy Orders' by superiors in Calcutta, who felt that ordination would entail social intercourse of a kind detrimental to British status in colonial society. Having been a 'subaltern' of mission for some twenty years, Goreh converted again, this time to High-Church Anglicanism. I demonstrate that he did this not only to secure his ordination (High-Church Anglicans being less averse to having Indian counterparts), but also because, in the process of understanding the faith he had embraced, he had become convinced by High-Church Tractarians of the “Grace of Orders.” I argue, therefore, that Goreh's little-known ordination quest demonstrates exemplary integrity, politically and theologically.
An investigation of two interventions for micrographia in individuals with Parkinson’s disease
Authors: Bryant MS , Rintala DH , Lai EC , Protas EJ 
Published: June 2010

Objective: This pilot study was designed to compare a change in micrographia between using grid lines and parallel horizontal lines as visual cues in individuals with Parkinson’s disease. Design: Single group pre- and post-test. Setting: Research lab. Participants: Eleven males with Parkinson’s disease. Interventions: Practice writing words with parallel and grid lines. The sequence of practising was randomized. Main outcome measures: Length of words. Results: The length of the words after practising with parallel lines was longer than in the initial free writing condition (17.83 ± 3.93 cm vs. 23.36 ± 5.82 cm, P =0.008). The length of the words after practising with grid lines was also longer than during free writing (17.83 ± 3.93 cm vs. 22.65 ± 4.04 cm, P =0.003). The length of the words after practising with parallel lines was not different from that after practising with grid lines. Conclusion: Improvements in letter size after practising with horizontal parallel lines and grid lines were not different. The addition of vertical lines to form a grid did not appear to improve the writing more than horizontal lines alone in persons with Parkinson’s disease who experienced micrographia.
Eyes on the prize: life goals in the context of visual disability in midlife
Authors: Popivker Luba , Wang Shu-wen , Boerner Kathrin 
Published: June 2010

Objective: To investigate implications of vision loss in midlife, identify life goals that are important to middle-aged adults and assess how vision loss interferes with goal pursuit. Design: Cross-sectional exploratory study. Setting: Vision rehabilitation agency. Subjects: Two hundred and sixteen middle-aged adults with visual impairment. Methods: Telephone interviews composed of structured and open-ended assessments of life goals (i.e. priorities, plans, or hopes people have in their lives) and goal interference due to vision loss. Results: Across the three assessed domains, functional goals were reported most often (N = 214), followed by social goals (N = 72) and psychological goals (N = 28). Among functional goals, career, daily tasks and mobility goals were identified by the highest percentage of participants. Family goals were identified most frequently for social goals, and life quality was identified most often for psychological goals. Vision-related goals occurred throughout the three domains, representing the majority of functional goals, a third of social goals, and almost half of psychological goals. Participants reported highest interference in functional and social goals and least in psychological goals. While participants reported moderate interference for most goals, markedly high interference was reported for daily tasks, mobility, independence and leisure goals. Goal identification was related to timing of onset of vision loss primarily for functional goals. Conclusions: Findings illuminate multifaceted goals held by middle-aged adults with vision impairment and how visual disability can interfere with goal pursuits. These findings suggest that identifying clients’ life goals and the vision-related interference they experience in goal pursuits may be a helpful step in vision rehabilitation services.