bluuebunny/crossref_metadata_2025 huggingface dataset:
Sustaining the State: Legal Consciousness and the Construction of Legality in Competing Abortion Activists' Narratives
Authors: Wilson Joshua C.
Published: May 2011

This article investigates how activists involved in both sides of the street politics of abortion simultaneously create, are constrained by, and use law when recounting a period of conflict that resulted in litigation. The activists‐turned‐litigants' construction of legality is explored by identifying and analyzing patterns of inclusion, absence, amendment, and type of law (i.e., state or extrastate) in and across the stories they tell. It is found that even though there are multiple reasons to expect all of these activists to resist or amend the state's conception of law, their narratives ultimately reproduce state law's legitimacy and power. The activists' stories also illustrate that legal consciousness is contextually and experientially based and is therefore subject to change. This finding has implications for legal mobilization as well as for the nature of legal consciousness.
Queer Legal History: A Field Grows Up and Comes Out
Authors: Kornbluh Felicia
Published: May 2011

This essay examines recent scholarship on the legal history of sexuality in the United States. It focuses on Margot Canaday's The Straight State: Sexuality and Citizenship in Modern America (2009) and Marc Stein's Sexual Injustice: Supreme Court Decisions from Griswold to Roe (2010). It also reviews recent work on the history of marriage, including Sarah Barringer Gordon's The Spirit of the Law: Religious Voices and the Constitution in Modern America (2010) and George Chauncey's Why Marriage? The History Shaping Today's Debate Over Gay Equality (2004), and the history of military law Defending America: Military Culture and the Cold‐War Court Martial (2005), by Elizabeth Lutes Hillman. The essay argues that this scholarship is significant because it offers a different view of sex and power than the one derived from the early writing of Michel Foucault. “Queer legal history” treats the liberalism of the 1960s‐1970s as sexually discriminatory as well as liberatory. It underlines the exclusions that were part of public policy under the federal G.I. Bill and the New Deal welfare state.
Body mass index and the incidence of influenza‐associated pneumonia in a UK primary care cohort
Authors: Blumentals William A. , Nevitt Alan , Peng Michael M. , Toovey Stephen
Published: May 2011

Please cite this paper as: Blumentals WA. et al. (2012) Body mass index and the incidence of influenza‐associated pneumonia in a UK primary care cohort. Influenza and Other Respiratory Viruses 6(1), 28–36.

Background  Accumulating data suggest an association between increased BMI/obesity and morbidity in patients with pandemic (H1N1) 2009 influenza. Information on metabolic status and prognosis in seasonal influenza is lacking, however.

Methods  A retrospective cohort study was carried out using the UK General Practice Research Database. Patients aged ≥18 with ≥1 recorded BMI in the 12–58 kg/m2 range between January 1, 2000, and December 31, 2007, were observed for an influenza‐associated pneumonia diagnosis after the date of baseline BMI, including ‘influenza with pneumonia’ or a diagnosis of ‘pneumonia’ up to 30 days after a diagnosis of ‘influenza’.

Results  A total of 1 074 315 patients were included, of whom 73·2% were within the reference BMI range or overweight and 2·2% were underweight (<18·5 kg/m2). Pneumonia rates were 32·33–37·48/100 000 in all BMI categories except the underweight (98·29/100 000). Relative to patients with acceptable weight, those who were underweight had an increased pneumonia rate [adjusted IRR = 2·32 (95% CI 1·80–2·94)], while being overweight (BMI = 25·0–29·9 kg/m2) or obese (BMI ≥ 30·0 kg/m2) was associated with a decreased pneumonia rate [adjusted IRR = 0·77 (95% CI 0·68–0·86) and 0·85 (95% CI 0·73–1·00), respectively]. On the other hand, women and obese women with type 2 diabetes had increased pneumonia rates [adjusted IRR = 1·37 (95% CI 1·08–1·72) and 1·47 (95%CI 1·01–2·06), respectively].

Conclusions  In contrast to initial data from pandemic influenza, influenza pneumonia, and pneumonia following influenza were the most common in underweight persons, and an apparent decreased rate of pneumonia was noted with increasing BMI categories. Women with type 2 diabetes had increased rates of pneumonia.
Messy Ethics: Conducting Moral Participatory Action Research in the Crucible of University–School Relations
Authors: Kuriloff Peter J. , Andrus Shannon H. , Ravitch Sharon M.
Published: May 2011

In this article we argue that when university researchers engage in democratic participatory action research with schools the process requires a special type of attention to the ethical difficulties which can arise. We note how current professional standards of ethics are inadequate to fully address many of the dilemmas faced in collaborative research. We then share examples of ethical dilemmas that have arisen in our work with schools and demonstrate how each has contributed to the (developing) framework we have created to avoid or manage the kinds of messy ethical issues we describe. We argue that this framework reflects a continuous commitment to an ethics of practice. We believe that those engaged in this type of work must assume an ethical stance and view all decisions in the research process as ethical ones that potentially affect the lives of all of those involved.
Bringing Research Into Educational Practice: Lessons Learned
Authors: Hille Katrin
Published: May 2011

Bringing research into educational practice is necessary but does not happen automatically. The Transfercenter for Neuroscience and Learning, at the University of Ulm in Germany, is set up to transfer (neuro)scientific knowledge into educational practice. In doing so we have learned why this does not happen automatically, and have tried to make sure it happens anyway. We have realized that transferring research into educational practice needs a special research approach, which we suggest could be labeled “translational research” as seen in medical research. We have also realized that transfer requires a special type of scientist. We try to hire generalists who work and think interdisciplinarily and who are committed to providing a service to practitioners. Finally, we feel that neuroscience provides a possible foundation for learning sciences.
Differential Susceptibility in Early Literacy Instruction Through Computer Games: The Role of the Dopamine D4 Receptor Gene (DRD4)
Authors: Kegel Cornelia A. T. , Bus Adriana G. , van IJzendoorn Marinus H.
Published: May 2011

Not every child seems equally susceptible to the same parental, educational, or environmental influences even if cognitive level is similar. This study is the first randomized controlled trial to apply the differential susceptibility paradigm to education in relation to children's genotype and early literacy skills. A randomized pretest–posttest control group design was used to examine the effects of the Intelligent Tutoring System Living Letters. Two intervention groups were created, 1 receiving feedback and 1 completing the program without feedback, and 1 control group. Carriers of the long variant of the dopamine D4 receptor gene (DRD4 7‐repeat) profited most from the computer program with positive feedback, whereas they performed at the lowest level of early literacy skills in the absence of such feedback. Our findings suggest that behind modest overall educational intervention effects a strong effect on a subgroup of susceptible children may be hidden.
Neural Differences in Bilingual Children's Arithmetic Processing Depending on Language of Instruction
Authors: Mondt Katrien , Struys Esli , Van den Noort Maurits , Balériaux Danielle , Metens Thierry , Paquier Philippe , Van de Craen Piet , Bosch Peggy , Denolin Vincent
Published: May 2011

Many children in bilingual regions follow lessons in a language at school (school‐language) that they hardly ever speak at home or in other informal settings. What are the neural effects of this phenomenon? This functional magnetic resonance imaging (fMRI) study investigates the effects of using school‐language on brain activity during a high cognitive task in two groups of French/Dutch bilingual children. Participants were asked to assess the correctness of 3‐operand (e.g., 5 − 2 = 3) and 4‐operand (e.g., 3 − 2 = 1 + 4) equations, respectively. Instructions about the task were given either in the children's school‐language or in another language that they only used at home. It was found that although both groups recruited a visuomotor occipitofrontal network in the left hemisphere, the children who performed the task in their school‐language showed less activation than the children who did not, indicating the importance of language of instruction for bilingual children's arithmetic processing.
Why Arts Integration Improves Long‐Term Retention of Content
Authors: Rinne Luke , Gregory Emma , Yarmolinskaya Julia , Hardiman Mariale
Published: May 2011

Advocates of the arts agree that the K‐12 curriculum should include dedicated time for arts instruction. Some have argued further that knowledge and skills acquired through the arts transfer to nonarts domains. Others claim that evidence of this kind of transfer is limited and instead argue that the arts cultivate valuable dispositions that help students succeed both in and outside of school. Another potential benefit of the arts has received little attention, however. Arts integration—the use of the arts as a teaching methodology throughout the curriculum—may improve long‐term retention of content. A variety of long‐term memory effects well known in cognitive psychology are reviewed, and it is argued that arts integration naturally takes advantage of these effects while promoting student motivation. This review of findings and applications provides an example of how existing research from neuroscience and cognitive science can inform the work of practicing educators.
Genetic polymorphisms in antioxidative enzymes are associated to forced expiratory volume in 1 s (FEV1) in smokers independently of asthma
Authors: Malling Tine Halsen , Sigsgaard Torben , Brasch‐Andersen Charlotte , Frischknecht Lone , Andersen Helle Raun , Kruse Torben Arvid , Sherson David , Skadhauge Lars Rauff , Thomsen Gert , Bælum Jesper , Omland Øyvind
Published: May 2011

Introduction:  In this study, we hypothesised that the genotypes coding for low antioxidative enzyme activity are associated with asthma and reduced lung function.

Methods:  Using the European Community Respiratory Health Survey protocol, we enlisted 1091 Danish subjects in this cross‐sectional study. Asthma phenotypes were defined as asthma symptoms in combination with steroid usage, bronchial hyperresponsiveness and atopy. These phenotypes and lung function were analysed with respect to glutathione peroxidase, GPX1 (Pro198Leu, rs1050450), manganese superoxide dismutase, SOD2 (Ala16Val, rs4880) and three glutathione S‐transferases; GSTP1 (Ile105Val, rs1695), GSTT1 (gene copy number) and GSTM1 (gene copy number).

Results:  We found no associations between these genotypes and the asthma phenotypes. For the 201 subjects identified as current smokers and recruited via random sampling, an association was seen between increasing number of genotypes coding for high antioxidative enzyme activity (GPX1 Pro/Pro, SOD2 Val/Val, GSTP1 Ile/Ile, GSTT1 two copies, GSTM1 two copies) and forced expiratory volume in 1 s (FEV1)% predicted. The increase in FEV1% predicted was 2.0% (95% confidence interval 0.3–3.8) per genotype. There was no identified significance for the inverse association between FEV1% predicted and number of genotypes coding for low antioxidative enzyme activity.

Conclusion:  The present study does not support the hypothesis that asthma is associated with genotypes of these major antioxidative enzymes. However, we speculate that since we see an impact of these genotypes on lung function in young adult smokers, polymorphisms in antioxidative enzymes may contribute to the range of susceptibility of smokers have to Chronic obstructive lung disease.

Please cite this paper as: Malling TH, Sigsgaard T, Brasch‐Andersen C, Frischknecht L, Andersen HR, Kruse TA, Sherson D, Skadhauge LR, Thomsen G, Bælum J and Omland Ø. Genetic polymorphisms in antioxidative enzymes are associated to forced expiratory volume in 1 s (FEV1) in smokers independently of asthma. Clin Respir J 2012; 6: 46–55.
Molecular profiling of diatom assemblages in tropical lake sediments using taxon‐specific PCR and Denaturing High‐Performance Liquid Chromatography (PCR‐DHPLC)
Authors: EPP LAURA S. , STOOF‐LEICHSENRING KATHLEEN R. , TRAUTH MARTIN H. , TIEDEMANN RALPH
Published: May 2011

Here we present a protocol to genetically detect diatoms in sediments of the Kenyan tropical Lake Naivasha, based on taxon‐specific PCR amplification of short fragments (approximately 100 bp) of the small subunit ribosomal (SSU) gene and subsequent separation of species‐specific PCR products by PCR‐based denaturing high‐performance liquid chromatography (DHPLC). An evaluation of amplicons differing in primer specificity to diatoms and length of the fragments amplified demonstrated that the number of different diatom sequence types detected after cloning of the PCR products critically depended on the specificity of the primers to diatoms and the length of the amplified fragments whereby shorter fragments yielded more species of diatoms. The DHPLC was able to discriminate between very short amplicons based on the sequence difference, even if the fragments were of identical length and if the amplicons differed only in a small number of nucleotides. Generally, the method identified the dominant sequence types from mixed amplifications. A comparison with microscopic analysis of the sediment samples revealed that the sequence types identified in the molecular assessment corresponded well with the most dominant species. In summary, the PCR‐based DHPLC protocol offers a fast, reliable and cost‐efficient possibility to study DNA from sediments and other environmental samples with unknown organismic content, even for very short DNA fragments.
Identifying the true oysters (Bivalvia: Ostreidae) with mitochondrial phylogeny and distance‐based DNA barcoding
Authors: LIU JUN , LI QI , KONG LINGFENG , YU HONG , ZHENG XIAODONG
Published: May 2011

Oysters (family Ostreidae), with high levels of phenotypic plasticity and wide geographic distribution, are a challenging group for taxonomists and phylogenetics. As a useful tool for molecular species identification, DNA barcoding might offer significant potential for oyster identification and taxonomy. This study used two mitochondrial fragments, cytochrome c oxidase I (COI) and the large ribosomal subunit (16S rDNA), to assess whether oyster species could be identified by phylogeny and distance‐based DNA barcoding techniques. Relationships among species were estimated by the phylogenetic analyses of both genes, and then pairwise inter‐ and intraspecific genetic divergences were assessed. Species forming well‐differentiated clades in the molecular phylogenies were identical for both genes even when the closely related species were included. Intraspecific variability of 16S rDNA overlapped with interspecific divergence. However, average intra‐ and interspecific genetic divergences for COI were 0–1.4% (maximum 2.2%) and 2.6–32.2% (minimum 2.2%), respectively, indicating the existence of a barcoding gap. These results confirm the efficacy of species identification in oysters via DNA barcodes and phylogenetic analysis.
Effective inhibition of melanoma by BI‐69A11 is mediated by dual targeting of the AKT and NF‐κB pathways
Authors: Feng Yongmei , Barile Elisa , De Surya K. , Stebbins John L. , Cortez Apple , Aza‐Blanc Pedro , Villanueva Jessie , Heryln Meenhard , Krajewski Stan , Pellecchia Maurizio , Ronai Ze’ev A. , Chiang Gary G.
Published: May 2011

In melanoma, the activation of pro‐survival signaling pathways, such as the AKT and NF‐κB pathways, is critical for tumor growth. We have recently reported that the AKT inhibitor BI‐69A11 causes efficient inhibition of melanoma growth. Here, we show that in addition to its AKT inhibitory activity, BI‐69A11 also targets the NF‐κB pathway. In melanoma cell lines, BI‐69A11 inhibited TNF‐α‐stimulated IKKα/β and IκB phosphorylation as well as NF‐κB reporter gene expression. Furthermore, the effective inhibition of melanoma growth by BI‐69A11 was attenuated upon NF‐κB activation. Mechanistically, reduced NF‐κB signaling by BI‐69‐A11 is mediated by the inhibition of sphingosine kinase 1, identified in a screen of 315 kinases. Significantly, we demonstrate that BI‐69A11 is well tolerated and orally active against UACC 903 and SW1 melanoma xenografts. Our results demonstrate that BI‐69A11 inhibits both the AKT and the NF‐κB pathways and that the dual targeting of these pathways may be efficacious as a therapeutic strategy in melanoma.
The phosphatidyl inositol 3‐kinase pathway is central to the pathogenesis of Kit‐activated melanoma
Authors: Liang Ruixia , Wallace Andrea R. , Schadendorf Dirk , Rubin Brian P.
Published: May 2011

Mouse Kit L575P, the ortholog of human KIT L576P, a common KIT mutation found in human melanoma was expressed in an immortalized but non‐transformed mouse Ink4a‐Arf‐deficient melanocyte cell line. The resultant Ink4a‐Arf‐deficient Kit L575P‐expressing melanocytes exhibited increased proliferation, the ability to grow in soft agar, and increased migration. When these cells were injected subcutaneously into NOD/SCID/gamma(c) mice, melanomas arose in 5 of 7 (71%) mice. One of seven mice (14%) injected with these cells developed metastatic disease. Evaluation of signal transduction pathways downstream of constitutively activated Kit L575P revealed striking activation of the phosphatidyl inositol 3‐kinase (PI3K) pathway. Inhibition of the PI3K pathway pharmacologically or genetically abolished the transformation phenotypes gained by the L575P single mutant. These studies validate this Kit L575P‐activated model of melanoma and establish the PI3K pathway as a dominant signaling pathway downstream of Kit in melanoma.
Variability and adaptability of Miscanthus species evaluated for energy crop domestication
Authors: YAN JUAN , CHEN WENLI , LUO FAN , MA HONGZHENG , MENG AIPING , LI XINWEI , ZHU MING , LI SHANSHAN , ZHOU HAIFEI , ZHU WEIXING , HAN BIN , GE SONG , LI JIANQIANG , SANG TAO
Published: May 2011

A growing body of evidence indicates that second‐generation energy crops can play an important role in the development of renewable energy and the mitigation of climate change. However, dedicated energy crops have yet to be domesticated in order to fully realize their productive potential under unfavorable soil and climatic conditions. To explore the possibility of domesticating Miscanthus crops in northern China where marginal and degraded land is abundant, we conducted common garden experiments at multiple locations to evaluate variation and adaptation of three Miscanthus species that are likely to serve as the wild progenitors of the energy crops. A total of 93 populations of Miscanthus sinensis, Miscanthus sacchariflorus, and Miscanthus lutarioriparius were collected across their natural distributional ranges in China and grown in three locations that represent temperate grassland with cold winter, the semiarid Loess Plateau, and relatively warm and wet central China. Evaluated with growth traits such as plant height, tiller number, tiller diameter, and flowering time, the Miscanthus species showed high levels of genetic variation within and between species. There were significant site × population interactions for almost all traits of M. sacchariflorus and M. sinensis, but not M. lutarioriparius. The northern populations of M. sacchariflorus had the highest establishment rates at the most northern site owing to their strong cold tolerance. An endemic species in central China, M. lutarioriparius, produced not only the highest biomass of the three species but also higher biomass at the Loess Plateau than the southern site near its native habitats. These results demonstrated that the wild species harbored a high level of genetic variation underlying traits important for crop establishment and production at sites that are colder and drier than their native habitats. The natural variation and adaptive plasticity found in the Miscanthus species indicated that they could provide valuable resources for the development of second‐generation energy crops.
Current drug discovery strategies for treatment of hepatitis C virus infection
Authors: Cheng K-C , Gupta Samir , Wang Hongwu , Uss Annette S , Njoroge George F , Hughes Eric
Published: May 2011

Objectives

Hepatitis C virus (HCV) infection represents a major worldwide-health problem. The current standard of care is combination therapy with pegylated interferon and ribavirin, which achieves a successful response in only approximately 40% of genotype I patients.

Key findings

The biology of HCV infection has been under intensive research and important progress has been made in understanding the replication cycle of the virus. Several therapeutic targets have been under investigation, such as NS3 protease, NS4A replicase and NS5B polymerase. New potential targets, such as NS2 protease, as well as CD-81 and claudin-1 entry co-receptors, have also been identified.

Clinical evaluations of drug candidates targeting NS3 protease, NS4A cofactor, and NS5B polymerase have demonstrated the potential of developing small molecules that interfere with the replication of the virus. Additional issues, including genotype coverage, resistant mutations, and combination therapy represent major challenges for future drug discovery efforts.
Gnaphaliin A and B relax smooth muscle of guinea-pig trachea and rat aorta via phosphodiesterase inhibition
Authors: Rodríguez-Ramos Fernando , González-Andrade Martín , Navarrete Andrés
Published: May 2011

Objectives

To explore the relaxant mechanism of action of gnaphaliin A and gnaphaliin B in guinea-pig trachea and rat aorta, and to investigate the theoretical and experimental phosphodiesterase (PDE) inhibitory activity of these flavones.

Methods

The relaxant effect and the inhibition of calcium chloride induced contractions of both flavones were evaluated on guinea-pig trachea and rat aorta rings. The PDE inhibitory activity was evaluated using a cyclic nucleotide PDE colorimetric assay kit with cAMP and cGMP as substrates. The docking analysis was carried out with AutoDock4 software and X-ray structure of PDE type 5. The activity of both gnaphaliins was compared with the activity of sildenafil, rolipram, aminophylline, IBMX and enoximone.

Key findings

Gnaphaliin A and B were more actives as relaxants on rat aorta than guinea-pig trachea. They were less potent in the relaxation of guinea-pig trachea and rat aorta than sildenafil, but they were equal or more potent than the other PDE inhibitors tested. The relaxant effect of these flavones was potentiated by nitroprusside and forskolin, and blocked by 1H-[1,2,4]-oxadiazolo[4,3-a]quinoxalin-1-one but not by 2′,5′-dideoxyadenosine in guinea-pig trachea. L-NAME did not modify the relaxant effect of gnaphaliins. Gnaphaliins were more potent as PDE inhibitors when cGMP was used as substrate. Docking analysis revealed that gnaphaiins bind to the same binding site of sildenafil at PDE type 5.

Conclusions

The results suggest that the main relaxant mechanism of action of gnaphaliin A and B is inhibition of PDEs with a preference to inhibit the degradation of cGMP. The docking study suggested that these flavones bind with high specificity to the same binding site of sildenafil at PDE type 5.
Metronidazole leads to enhanced uptake of imatinib in brain, liver and kidney without affecting its plasma pharmacokinetics in mice
Authors: Tan Shin Yee , Kan Elaine , Lim Wei Yin , Chay Grace , Law Jason H K , Soo Gian Wan , Bukhari Nadeem Irfan , Segarra Ignacio
Published: May 2011

Objectives

The pharmacokinetic interaction between metronidazole, an antibiotic–antiparasitic drug used to treat anaerobic bacterial and protozoal infections, and imatinib, a CYP3A4, P-glycoprotein substrate kinase inhibitor anticancer drug, was evaluated.

Methods

Male imprinting control region mice were given 50 mg/kg imatinib PO (control group) or 50 mg/kg imatinib PO, 15 min after 40 mg/kg PO metronidazole (study group). Imatinib plasma, brain, kidney and liver concentrations were measured by HPLC and non-compartmental pharmacokinetic parameters estimated.

Key findings

Metronidazole coadministration resulted in a double-peak imatinib disposition profile. The maximum concentration (Cmax) decreased by 38%, the area under the curve (AUC0–∞) decreased by 14% and the time to Cmax (Tmax) was earlier (50%) in plasma. Apparent volume of distribution (VSS/F) and oral clearance (Cl/F) increased by 21% and 17%, respectively. Imatinib tissue penetration was higher after metronidazole coadministration, with 1.7 and 2.1-fold AUC0–∞ increases in liver and kidney, respectively. Metronidazole increased imatinib's tissue-to-plasma AUC0–∞ ratio in liver from 2.29 to 4.53 and in kidney from 3.04 to 7.57, suggesting higher uptake efficiency. Brain Cmax was 3.9-fold higher than control and AUC0–t last was 2.3-fold greater than plasma (3.5% in control group). No tissue-plasma concentration correlation was found.

Conclusions

Metronidazole slightly decreased imatinib systemic exposure but enhanced liver, kidney and brain penetration, probably due to metronidazole-mediated inhibition of P-glycoprotein and other efflux transporters. The high brain exposure opens possibilities for treatment of glioma and glioblastoma. Renal and hepatic functions may need to be monitored due to potential renal and hepatic toxicity.
Oral and Nasal DNA Vaccines Delivered by Attenuated Salmonella enterica Serovar Typhimurium Induce a Protective Immune Response against Infectious Bronchitis in Chickens
Authors: Jiao Hongmei , Pan Zhiming , Yin Yuelan , Geng Shizhong , Sun Lin , Jiao Xinan
Published: May 2011

Several studies have reported that intramuscular injection of DNA vaccines against infectious bronchitis virus (IBV) induces protective immune responses. In the present study, we developed oral and nasal DNA vaccines that carried the S1 gene and N gene of IBV delivered by attenuatedSalmonella entericaserovar Typhimurium strains SL/pV-S1 and SL/pV-N, respectively. The safety and stability of recombinantSalmonellavaccine were evaluated. Following oral and nasal administration to chickens, the serum and mucosal samples were collected and antibodies against IBV were measured. Chickens were then challenged with IBV strain M41 by the nasal-ocular route 3 weeks after boosting. The results showed that oral and nasal immunization with coadministered SL/pV-S1 and SL/pV-N elicited significant IBV-specific humoral and mucosal immune responses and conferred protective efficacy against IBV challenge higher than that in chickens immunized only with SL/pV-S1. The current study shows that novel DNA vaccines delivered by attenuatedS.Typhimurium may be promising candidates for the prevention of infectious bronchitis (IB).These vaccines are efficacious, easily produced economically, and able to be delivered orally and nasally rather than injected. Coadministration of SL/pV-S1 and SL/pV-N may represent an effective mucosal vaccination regimen.
Within-Subject Variability of Mycobacterium tuberculosis-Specific Gamma Interferon Responses in German Health Care Workers
Authors: Ringshausen Felix C. , Nienhaus Albert , Torres Costa José , Knoop Heiko , Schlösser Stephan , Schultze-Werninghaus Gerhard , Rohde Gernot
Published: May 2011

Gamma interferon (IFN-γ) release assays (IGRAs) are used increasingly for the periodic tuberculosis (TB) screening of health care workers (HCWs), although data regarding the reproducibility and interpretation of serial testing results in countries with a low incidence of TB are scarce. The present study evaluated and compared the within-subject variability of dichotomous and continuous results of two commercial IGRAs, the QuantiFERON-TB Gold In-Tube (QFT) and the T-SPOT.TB (T-SPOT), in German HCWs during a 4-week period. Thirty-five immunocompetent HCWs with low or medium TB screening risk and without known recent TB exposure or tuberculin skin test application were tested repeatedly with both IGRAs at weekly intervals. A total of 158 valid results were obtained for each IGRA. Changes of about ±70% (QFT) and ±60% (T-SPOT) from the mean IFN-γ response accounted for 95% of the within-subject variability. However, according to the manufacturers' cutoffs, inconsistent results were observed more frequently for the QFT (28.6%; four conversions, six reversions) than for the T-SPOT (8.6%; three reversions;P< 0.001). The overall agreement between the IGRAs was good. Regression toward the means accounted for a significant decline in mean IFN-γ responses of about 25% between successive visits for both IGRAs. Although both assays were highly reliable and reproducible, we observed substantial within-subject variability and regression toward the means during a 4-week period, which should be considered when interpreting serial testing results in comparable populations and settings. Our data support the use of borderline zones for the interpretation of serial IGRA results and the retesting of borderline positive results before offering preventive chemotherapy.
Rapid Identification of Cryptococcus neoformans var. grubii, C. neoformans var. neoformans, and C. gattii by Use of Rapid Biochemical Tests, Differential Media, and DNA Sequencing
Authors: McTaggart Lisa , Richardson Susan E. , Seah Christine , Hoang Linda , Fothergill Annette , Zhang Sean X.
Published: May 2011

Rapid identification of
Cryptococcus neoformans
var.
grubii
,
Cryptococcus neoformans
var.
neoformans
, and
Cryptococcus gattii
is imperative for facilitation of prompt treatment of cryptococcosis and for understanding the epidemiology of the disease. Our purpose was to evaluate a test algorithm incorporating commercial rapid biochemical tests, differential media, and DNA sequence analysis that will allow us to differentiate these taxa rapidly and accurately. We assessed 147 type, reference, and clinical isolates, including 6 other
Cryptococcus
spp. (10 isolates) and 14 other yeast species (24 isolates), using a 4-hour urea broth test (Remel), a 24-hour urea broth test (Becton Dickinson), a 4-hour caffeic acid disk test (Hardy Diagnostics and Remel), 40- to 44-hour growth assessment on
l
-canavanine glycine bromothymol blue (CGB) agar, and intergenic spacer (IGS) sequence analysis. All 123
Cryptococcus
isolates hydrolyzed urea, along with 7 isolates of
Rhodotorula
and
Trichosporon
. Eighty-five of 86
C. neoformans
(99%) and 26 of 27
C. gattii
(96%) isolates had positive caffeic acid results, unlike the other cryptococci (0/10) and yeast species (0/24). Together, these two tests positively identified virtually all
C. neoformans/C. gattii
isolates (98%) within 4 h. CGB agar or IGS sequencing further differentiated these isolates within 48 h. On CGB, 25 of 27 (93%)
C. gattii
strains induced a blue color change, in contrast to 0 of 86
C. neoformans
isolates. Neighbor-joining cluster analysis of IGS sequences differentiated
C. neoformans
var.
grubii
,
C. neoformans
var.
neoformans
, and
C. gattii
. Based on these results, we describe a rapid identification algorithm for use in a microbiology laboratory to distinguish clinically relevant
Cryptococcus
spp.
Isolation of Nocardia beijingensis from a Pulmonary Abscess Reveals Human Immunodeficiency Virus Infection
Authors: Martinaud Christophe , Verdonk Charles , Bousquet Aurore , MacNab Christine , Vaylet Fabien , Soler Charles , Samson Thierry , Margery Jacques
Published: May 2011

A strain of
Nocardia
was isolated from a pulmonary abscess of a human immunodeficiency virus-infected patient in France. Comparative 16S rRNA gene sequence analysis revealed that the isolate represented a strain of
Nocardia beijingensis
. Antimicrobial susceptibility testing was essential to guide the clinicians to successfully treat this infection.
Acute Necrotizing Sinusitis Caused by Staphylococcus lugdunensis
Authors: Matthews Philippa C. , Lazarus Rajeka , Protheroe Andrew , Milford Christopher , Bowler Ian C. J. W.
Published: May 2011

Staphylococcus lugdunensis
is most commonly associated with infections arising from the inguinal region, but here we report this organism as a cause of bacterial sinusitis, highlighting its potential niche as a commensal of the upper airways. The severity of necrosis demonstrates the potential for destructive pathology mimicking
Staphylococcus aureus
disease.
Xpert MTB/RIF: a New Pillar in Diagnosis of Extrapulmonary Tuberculosis?
Authors: Vadwai Viral , Boehme Catharina , Nabeta Pamela , Shetty Anjali , Alland David , Rodrigues Camilla
Published: May 2011

Approximately 10 to 15% of tuberculosis (TB) cases in India are estimated to have extrapulmonary disease, and due to a lack of diagnostic means, they often remain untreated. The early detection of
Mycobacterium tuberculosis
and multidrug resistance is a priority in TB diagnosis to improve the successful treatment rate of TB and reduce transmission. The Xpert MTB/RIF (Xpert) test, recently endorsed by the World Health Organization for the detection of pulmonary TB, was evaluated to test its utility in 547 patients with suspected extrapulmonary tuberculosis. Five hundred forty-seven extrapulmonary specimens were split and processed simultaneously for both culture (solid and liquid) and Xpert testing. For culture, the sensitivity was low, 53% (150/283 specimens). Xpert sensitivity and specificity results were assessed in comparison to a composite reference standard made up of smear and culture results and clinical, radiological, and histological findings. The sensitivity of the Xpert assay was 81% (228/283 specimens) (64% [89/138] for smear-negative cases and 96% [139/145] for smear-positive cases), with a specificity of 99.6%. The sensitivity was found to be high for the majority of specimen types (63 to 100%) except for cerebrospinal fluid, the sensitivity of which was 29% (2/7 specimens). The Xpert test correctly identified 98% of phenotypic rifampin (RIF)-resistant cases and 94% of phenotypic RIF-susceptible cases. Sequencing of the 6 discrepant samples resolved 3 of them, resulting in an increased specificity of 98%. In conclusion, the results of this study suggest that the Xpert test also shows good potential for the diagnosis of extrapulmonary TB and that its ease of use makes it applicable for countries where TB is endemic.
Design and Performance of the CDC Real-Time Reverse Transcriptase PCR Swine Flu Panel for Detection of 2009 A (H1N1) Pandemic Influenza Virus
Authors: Shu Bo , Wu Kai-Hui , Emery Shannon , Villanueva Julie , Johnson Roy , Guthrie Erica , Berman LaShondra , Warnes Christine , Barnes Nathelia , Klimov Alexander , Lindstrom Stephen
Published: May 2011

Swine influenza viruses (SIV) have been shown to sporadically infect humans and are infrequently identified by the Influenza Division of the Centers for Disease Control and Prevention (CDC) after being received as unsubtypeable influenza A virus samples. Real-time reverse transcriptase PCR (rRT-PCR) procedures for detection and characterization of North American lineage (N. Am) SIV were developed and implemented at CDC for rapid identification of specimens from cases of suspected infections with SIV. These procedures were utilized in April 2009 for detection of human cases of 2009 A (H1N1) pandemic (pdm) influenza virus infection. Based on genetic sequence data derived from the first two viruses investigated, the previously developed rRT-PCR procedures were optimized to create the CDC rRT-PCR Swine Flu Panel for detection of the 2009 A (H1N1) pdm influenza virus. The analytical sensitivity of the CDC rRT-PCR Swine Flu Panel was shown to be 5 copies of RNA per reaction and 10
−1.3∼−0.7
50% infectious doses (ID
50
) per reaction for cultured viruses. Cross-reactivity was not observed when testing human clinical specimens or cultured viruses that were positive for human seasonal A (H1N1, H3N2) and B influenza viruses. The CDC rRT-PCR Swine Flu Panel was distributed to public health laboratories in the United States and internationally from April 2009 until June 2010. The CDC rRT-PCR Swine Flu Panel served as an effective tool for timely and specific detection of 2009 A (H1N1) pdm influenza viruses and facilitated subsequent public health response implementation.
Antiviral Drug Research Proposal Activity
Authors: Injaian Lisa , Smith Ann C. , Shipley Jennifer German , Marbach-Ad Gili , Fredericksen Brenda
Published: May 2011

The development of antiviral drugs provides an excellent example of how basic and clinical research must be used together in order to achieve the final goal of treating disease. A Research Oriented Learning Activity was designed to help students to better understand how basic and clinical research can be combined toward a common goal. Through this project students gained a better understanding of the process of scientific research and increased their information literacy in the field of virology. The students worked as teams to research the many aspects involved in the antiviral drug design process, with each student becoming an “expert” in one aspect of the project. The Antiviral Drug Research Proposal (ADRP) culminated with students presenting their proposals to their peers and local virologists in a poster session. Assessment data showed increased student awareness and knowledge of the research process and the steps involved in the development of antiviral drugs as a result of this activity.
The Genetic Bottleneck in Vertical Transmission of Subtype C HIV-1 Is Not Driven by Selection of Especially Neutralization-Resistant Virus from the Maternal Viral Population
Authors: Russell Elizabeth S. , Kwiek Jesse J. , Keys Jessica , Barton Kirston , Mwapasa Victor , Montefiori David C. , Meshnick Steven R. , Swanstrom Ronald
Published: May 2011

Subtype C human immunodeficiency virus type 1 (HIV-1C) continues to cause the majority of new cases of mother-to-child transmission (MTCT), and yet there are limited data on HIV-1C transmission. We amplified
env
from plasma RNA for 19 HIV-1C MTCT pairs, 10 transmitting
in utero
(IU) and 9 transmitting intrapartum (IP). There was a strong genetic bottleneck between all mother-infant pairs, with a majority of transmission events involving the transmission of a single virus.
env
genes of viruses transmitted to infants IP, but not IU, encoded Env proteins that were shorter and had fewer putative N-linked glycosylation sites in the V1-V5 region than matched maternal sequences. Viruses pseudotyped with
env
clones representative of each maternal and infant population were tested for neutralization sensitivity. The 50% inhibitory concentration of autologous serum was similar against both transmitted (infant) and nontransmitted (maternal) viruses in a paired analysis. Mother and infant Env proteins were also similar in sensitivity to soluble CD4, to a panel of monoclonal antibodies, and to heterologous HIV-1C sera. In addition, there was no difference in the breadth or potency of neutralizing antibodies between sera from 50 nontransmitting and 23 IU and 23 IP transmitting HIV-1C-infected women against four Env proteins from heterologous viruses. Thus, while a strong genetic bottleneck was detected during MCTC, with viruses of shorter and fewer glycosylation sites in
env
present in IP transmission, our data do not support this bottleneck being driven by selective resistance to antibodies.
Ex Vivo
Bioluminescence Detection of Alcelaphine Herpesvirus 1 Infection during Malignant Catarrhal Fever
Authors: Dewals Benjamin , Myster Françoise , Palmeira Leonor , Gillet Laurent , Ackermann Mathias , Vanderplasschen Alain
Published: May 2011

Alcelaphine herpesvirus 1
(AlHV-1), carried by wildebeest asymptomatically, causes malignant catarrhal fever (WD-MCF) when cross-species transmitted to a variety of susceptible species of the
Artiodactyla
order. Experimentally, WD-MCF can be reproduced in rabbits. WD-MCF is described as a combination of lymphoproliferation and degenerative lesions in virtually all organs and is caused by unknown mechanisms. Recently, we demonstrated that WD-MCF is associated with the proliferation of CD8
+
cells supporting a latent type of infection in lymphoid tissues. Here, we investigated the macroscopic distribution of AlHV-1 infection using
ex vivo
bioluminescence imaging in rabbit to determine whether it correlates with the distribution of lesions in lymphoid and nonlymphoid organs. To reach that goal, a recombinant AlHV-1 strain was produced by insertion of a luciferase expression cassette (
luc
) in an intergenic region.
In vitro
, the reconstituted AlHV-1
luc

+

strain replicated comparably to the parental strain, and luciferase activity was detected by bioluminescence imaging.
In vivo
, rabbits infected with the AlHV-1
luc

+

strain developed WD-MCF comparably to rabbits infected with the parental wild-type strain, with hyperthermia and increases of both CD8
+
T cell frequencies and viral genomic charge over time in peripheral blood mononuclear cells and in lymph nodes at time of euthanasia. Bioluminescent imaging revealed that AlHV-1 infection could be detected
ex vivo
in lymphoid organs but also in lung, liver, and kidney during WD-MCF, demonstrating that AlHV-1 infection is prevalent in tissue lesions. Finally, we show that the infiltrating mononuclear leukocytes in nonlymphoid organs are mainly CD8
+
T cells and that latency is predominant during WD-MCF.
The Internal Kaposi's Sarcoma-Associated Herpesvirus LANA Regions Exert a Critical Role on Episome Persistence
Authors: De León Vázquez Erika , Kaye Kenneth M.
Published: May 2011

Kaposi's sarcoma-associated herpesvirus (KSHV) latency-associated nuclear antigen (LANA) is a 1,162-amino-acid protein that acts on viral terminal repeat (TR) DNA to mediate KSHV episome persistence. The two essential components of episome persistence are DNA replication prior to cell division and episome segregation to daughter nuclei. These functions are located within N- and C-terminal regions of LANA. N- and C-terminal regions of LANA are sufficient for TR DNA replication. In addition, N- and C-terminal regions of LANA tether episomes to mitotic chromosomes to segregate episomes to progeny cell nuclei. To generate a tethering mechanism, N-terminal LANA binds histones H2A/H2B to attach to mitotic chromosomes, and C-terminal LANA binds TR DNA and also associates with mitotic chromosomes. Here, we test the importance of the internal LANA sequence for episome persistence. We generated LANA mutants that contain N- and C-terminal regions of LANA but have most of the internal sequence deleted. As expected, the LANA mutants bound mitotic chromosomes in a wild-type pattern and also bound TR DNA as assayed by electrophoretic mobility shift assays (EMSA). The mutants mediated TR DNA replication, although with reduced efficiency compared with LANA. Despite the ability to replicate DNA and exert the chromosome and DNA binding functions necessary for segregating episomes to daughter nuclei, the mutants were highly deficient for the ability to mediate both short- and long-term episome persistence. These data indicate that internal LANA sequence exerts a critical effect on its ability to maintain episomes, possibly through effects on TR DNA replication.
Cyclophilin A Interacts with Domain II of Hepatitis C Virus NS5A and Stimulates RNA Binding in an Isomerase-Dependent Manner
Authors: Foster Toshana L. , Gallay Philippe , Stonehouse Nicola J. , Harris Mark
Published: May 2011

NS5A plays a critical, yet poorly defined, role in hepatitis C virus genome replication. The protein consists of three domains, each of which is able to bind independently to the 3′ untranslated region (UTR) of the viral positive strand genomic RNA. The peptidyl-prolyl isomerase cyclophilin A (CypA) binds to domain II, catalyzingcis-transisomerization. CypA inhibitors such as cyclosporine (CsA) have been shown to inhibit hepatitis C virus (HCV) replication. We show here that CypA stimulated domain II RNA binding activity, and this stimulation was abrogated by CsA. An isomerase mutant of CypA (H126Q) failed to bind to domain II and did not stimulate RNA binding. Finally, we demonstrate that the RNA binding of two domain II mutants, the D316E and D316E/Y317N mutants, previously shown to exhibit CypA independence for RNA replication, was unaffected by CypA. This study provides an insight into the molecular mechanism of CypA activity during HCV replication and further validates the use of CypA inhibitors in HCV therapy.
The Virulence of 1997 H5N1 Influenza Viruses in the Mouse Model Is Increased by Correcting a Defect in Their NS1 Proteins
Authors: Spesock April , Malur Meghana , Hossain M. Jaber , Chen Li-Mei , Njaa Bradley L. , Davis Charles T. , Lipatov Aleksandr S. , York Ian A. , Krug Robert M. , Donis Ruben O.
Published: May 2011

The NS1 protein of human influenza A viruses binds the 30-kDa subunit of the cleavage and polyadenylation specificity factor (CPSF30), a protein required for 3′ end processing of cellular pre-mRNAs, thereby inhibiting production of beta interferon (IFN-β) mRNA. The NS1 proteins of pathogenic 1997 H5N1 viruses contain the CPSF30-binding site but lack the consensus amino acids at positions 103 and 106, F and M, respectively, that are required for the stabilization of CPSF30 binding, resulting in nonoptimal CPSF30 binding in infected cells. Here we have demonstrated that strengthening CPSF30 binding, by changing positions 103 and 106 in the 1997 H5N1 NS1 protein to the consensus amino acids, results in a remarkable 300-fold increase in the lethality of the virus in mice. Unexpectedly, this increase in virulence is not associated with increased lung pathology but rather is characterized by faster systemic spread of the virus, particularly to the brain, where increased replication and severe pathology occur. This increased spread is associated with increased cytokine and chemokine levels in extrapulmonary tissues. We conclude that strengthening CPSF30 binding by the NS1 protein of 1997 H5N1 viruses enhances virulence in mice by increasing the systemic spread of the virus from the lungs, particularly to the brain.
Extensive Geographical Mixing of 2009 Human H1N1 Influenza A Virus in a Single University Community
Authors: Holmes Edward C. , Ghedin Elodie , Halpin Rebecca A. , Stockwell Timothy B. , Zhang Xing-Quan , Fleming Regina , Davey Richard , Benson Constance A. , Mehta Sanjay , Taplitz Randy , Liu Yu-Tseung , Brouwer Kimberly C. , Wentworth David E. , Lin Xudong ,   the INSIGHT FLU002 Study Group, Schooley Robert T.
Published: May 2011

Despite growing interest in the molecular epidemiology of influenza virus, the pattern of viral spread within individual communities remains poorly understood. To determine the phylogeography of influenza virus in a single population, we examined the spatial diffusion of H1N1/09 influenza A virus within the student body of the University of California, San Diego (UCSD), sampling for a 1-month period between October and November 2009. Despite the highly focused nature of our study, an analysis of complete viral genome sequences revealed between 24 and 33 independent introductions of H1N1/09 into the UCSD community, comprising much of the global genetic diversity in this virus. These data were also characterized by a relatively low level of on-campus transmission as well as extensive spatial mixing, such that there was little geographical clustering by either student residence or city ZIP code. Most notably, students experiencing illness on the same day and residing in the same dorm possessed phylogenetically distinct lineages. H1N1/09 influenza A virus is therefore characterized by a remarkable spatial fluidity, which is likely to impede community-based methods for its control, including class cancellations, quarantine, and chemoprophylaxis.