bluuebunny/crossref_metadata_2025 huggingface dataset:
Organizational climate in primary care settings: Implications for nurse practitioner practice
Authors: Poghosyan Lusine , Nannini Angela , Clarke Sean
Published: August 2012

Purpose

The purpose of this review is to investigate literature related to organizational climate, define organizational climate, and identify its domains for nurse practitioner (NP) practice in primary care settings.

Data sources

A search was conducted using MEDLINE, PubMed, HealthSTAR/Ovid, ISI Web of Science, and several other health policy and nursingy databases.

Conclusions

In primary care settings, organizational climate for NPs is a set of organizational attributes, which are perceived by NPs about their practice setting, emerge from the way the organization interacts with NPs, and affect NP behaviors and outcomes. Autonomy, NP–physician relations, and professional visibility were identified as organizational climate domains.

Implications for practice

NPs should be encouraged to assess organizational climate in their workplace and choose organizations that promote autonomy, collegiality between NPs and physicians, and encourage professional visibility. Organizational and NP awareness of qualities that foster NP practice will be a first step for developing strategies to creating an optimal organizational climate for NPs to deliver high‐quality care. More research is needed to develop a comprehensive conceptual framework for organizational climate and develop new instruments to accurately measure organizational climate and link it to NP and patient outcomes.
New Products and Corruption: Evidence from Indian Firms
Authors: de Waldemar Felipe Starosta
Published: August 2012

It has been shown that corruption has a negative effect on firm growth, but what about its impact on product innovation? I find that corruption, functioning as a bribe tax, diminishes the probability of new products being introduced. I use a World Bank Enterprise Survey conducted in India in 2005, with 1,600 firms answering both whether they introduced a new product to the firm, and whether and how much was paid in bribes. Controlling for innovation determinants and firm characteristics, sector‐state bribery averages have a negative and significant impact on product innovation.
Estimation of Public–Private Wage Differentials in Turkey with Sample Correction
Authors: San Sayın , Polat Özgür
Published: August 2012

The aim of this study is to determine how the change in the balance between public–private sector employments affected public and private earnings during the 1990s and 2000s in Turkey. We use the Oaxaca–Blinder and quantile decomposition methods to determine the wage gap between public and private sectors utilizing the 1994 Household Income Distribution and Consumption Expenditure Survey and the 2008 Household Budget Survey conducted by the Turkish Statistical Institute. The study determined that the primary difference in the average wages between sectors arises from the differences in the endowments without correction for gender. After adjusting for correction using quantile regression, we find that the difference in the endowments between sectors at lower quantiles explains the majority of the raw wage gap; whereas a substantial amount of the raw wage gap is explained by the sector effect at higher quantiles.
Experiences of algal bloom control using green solutions barley straw and ultrasound, an industry perspective
Authors: Purcell Diane , Parsons Simon A. , Jefferson Bruce , Holden Stuart , Campbell Andrew , Wallen Anna , Chipps Michael , Holden Barry , Ellingham Adam
Published: August 2012

Water companies in the United Kingdom have been dealing with seasonal algal blooms for many decades, with escalating costs as bloom occurrences have increased markedly with elevated demand on water supply. Environmentally friendly water treatment solutions, such as ultrasound and barley straw, are defined as those that use less energy and chemicals. Barley straw has been used quite extensively by certain UK water companies, and results are generally positive. Ultrasound has only recently been used as a treatment for algal removal, with limited results from several successful trials finding cyanobacteria to be the most susceptible algal group at field scale, with results also indicating susceptibility in other algal groups; however, energy costs are higher than barley straw. Ultrasound has been extremely successful at removing bacteria in wastewater treatment; therefore, ultrasound should be considered as an in‐works solution where its effectiveness would be enhanced while reducing energy requirements.
Impact of source waters, disinfectants, seasons and treatment approaches on trihalomethanes in drinking water: a comparison based on the size of municipal systems
Authors: Chowdhury Shakhawat
Published: August 2012

This study compares concentrations of trihalomethanes (THMs) in municipal water for 2001–2007 from the small and large systems in two provinces in Canada (Newfoundland and Quebec) based on source waters, disinfectants, seasons and treatment approaches. Approximately 71 and 94%, respectively, of the municipal systems in Quebec and Newfoundland are small systems (serving fewer than 3000 people). The small systems serve approximately 8.6% (0.57 million) and 44.1% (0.18 million) of the populations in Quebec and Newfoundland, respectively. Concentrations of THMs and its variability are much higher in the small systems (Quebec: 0–941 μg/L; Newfoundland: 0–875 μg/L) than in the systems with populations 10 000 or more (Quebec: 0–364 μg/L; Newfoundland: 2.3–205 μg/L). The study reveals that the differences in THMs between the small and medium/large systems are because of different types of source waters, treatments, disinfection strategies and seasons. The results emphasize that regulatory agencies must focus more on the occurrence of DBPs in small systems and identify strategies to reduce their levels in drinking water.
Investigation of the relation between meteorological parameters, North Atlantic Oscillation and groundwater levels in Torbali Region, Turkey
Authors: Dogan Mustafa , Ozgenc Aksoy Aysegul
Published: August 2012

Groundwater, the most reliable fresh water source, is used for drinking, domestic and agricultural purposes. Thereby, understanding its behaviour is important for the sustainability of groundwater sources. In this study, relations between the trends of climate parameters [recharge depending on precipitation, temperature and North Atlantic Oscillation Index (NAOI)] and groundwater levels trend were investigated for Torbali Region in Turkey. The human impact that is one of the deterministic components on the groundwater level has been removed from the groundwater level data sets. An increasing trend was observed in the early 1990s, and turning points were determined by using paired t‐test. The trends of the groundwater levels indicate that climate parameters affect groundwater levels in the similar manner. According to the results of the analyses, it is revealed that there is a similar linear variation that is strong and inverse between the trends of NAOI and meteorological indicator (temperature and recharge) and groundwater level.
Sensory Attribute Preservation in Extra Virgin Olive Oil with Addition of Oregano Essential Oil as Natural Antioxidant
Authors: Asensio Claudia M. , Nepote Valeria , Grosso Nelson R.
Published: August 2012

Abstract:  Four commercial varieties of oregano are farmed in Argentina: “Compacto,”“Cordobes,”“Criollo,” y “Mendocino.” Oregano essential oil is known for antioxidant properties. The objective of this study was to evaluate changes in the intensities of positive and negative attributes in extra virgin olive oil with addition of essential oil obtained from the 4 Argentinean oregano types. Oregano essential oil was added into olive oil at 0.05% w/w. The samples were stored in darkness and light exposure during 126 d at room temperature. The intensity ratings of fruity, pungency, bitterness, oregano flavor, and rancid flavor were evaluated every 21 d by a trained sensory panel. In general, samples with addition of oregano essential oil in olive oil exhibited higher and lower intensity ratings of positive and negative attributes, respectively, during storage compared with the control samples. The first 2 principal components explained 72.3% of the variability in the olive oil samples. In general, positive attributes of olive oil were highly associated with the addition of oregano essential oil in darkness, whereas rancid flavor was negatively associated with them. Olive oil with oregano “Cordobes” essential oil was oppositely associated with light exposure treatments and negative attribute (rancid flavor) suggesting better performance as natural antioxidant of this essential oil in olive oil. The result of this study showed that the presence of oregano essential oil, specially “Cordobes” type, preserve sensory quality of extra virgin olive oil prolonging the shelf life of this product.

Practical Application:  Extra virgin olive oil is highly appreciated for its health benefits, taste, and aroma. These properties are an important aspect in this product quality and need to be preserved. The addition of natural additives instead of synthetic ones covers the present trend in food technology. This research showed that the addition of oregano essential oil preserved the intensity ratings of positive attributes in extra virgin olive oil during storage. The essential oil of the oregano variety called “Cordobes” exhibited better protecting effect on sensory properties of olive oil than the other oregano varieties. The addition of oregano essential oil should be considered for the food industry as a natural source of antioxidant additives for preserving sensory properties in extra virgin olive oil and other similar food products.
Concentrations and Bioconcentration Factors of Minerals in Yellow‐Cracking Bolete (Xerocomus Subtomentosus) Mushroom Collected in Noteć Forest, Poland
Authors: Jarzyńska Grażyna , Chojnacka Aleksandra , Dryżałowska Anna , Nnorom Innocent C. , Falandysz Jerzy
Published: August 2012

Abstract:  Yellow‐cracking Bolete (Xerocomus subtomentosus) mushrooms and soil were collected from Noteć Forest–a large forested enclave in western part of Poland. Mercury was determined by cold vapour atomic absorption spectroscopy and the other elements by inductively coupled plasma atomic emission spectroscopy. K, P, and Mg were particularly abundant, with mean values of 46000, 8400, and 1100 mg/kg dry weight (dw) in caps followed by Na, Rb, Zn, and Ca with mean concentrations of 580, 350, 200, and 170 mg/kg dw, respectively. In descending order, the mean concentrations of Fe, Al, Cu, and Mn were 52, 49, 46, and 14 mg/kg dw, while the mean for the remaining elements was around 1.0 mg/kg dw or less. The elements such as Ca, Cu, Hg, K, Mg, Na, P, Rb, Zn, Ag, Cd, and Ni were accumulated (with bioconcentration factor (BCF) > 1), while Al, Ba, Fe, Mn, Sr, Co, Cr, and Pb were excluded (BCF < 1) in the fruiting bodies. The Pb and Cd content did not exceed the maximum levels set by the EU for cultivated mushrooms. Mercury in a conventional meal (300 g) portion of Yellow‐cracking Bolete was far below the provisionally tolerable weekly intake of 0.004 mg/kg body weight (bw) as reevaluated recently by WHO.

Practical Application:  The method presented in this study allows one to determine the content of 20 elements (Ag, Al, Ba, Ca, Cd, Co, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Ni, P, Pb, Rb, Sr, and Zn) in caps and stipes of Yellow‐cracking Bolete (Xerocomus subtomentosus) mushrooms and soil samples collected from Poland. This study has revealed that the total Cd, Hg, and Pb dose provided to human body due to consumption of Yellow‐cracking Bolete does not pose threat to a consumer's health.
Comparison of Decontamination Efficacy of Antimicrobial Treatments for Beef Trimmings against Escherichia coli O157:H7 and 6 Non‐O157 Shiga Toxin‐Producing E. coli Serogroups
Authors: Geornaras Ifigenia , Yang Hua , Manios Stavros , Andritsos Nikolaos , Belk Keith E. , Nightingale Kendra K. , Woerner Dale R. , Smith Gary C. , Sofos John N.
Published: August 2012

Abstract:  The decontamination efficacy of 6 chemical treatments for beef trimmings were evaluated against Escherichia coli O157:H7 and 6 non‐O157 Shiga toxin‐producing E. coli (nSTEC) serogroups. Rifampicin‐resistant 4‐strain mixtures of E. coli O157:H7 and nSTEC serogroups O26, O45, O103, O111, O121, and O145 were separately inoculated (3 to 4 log CFU/cm2) onto trimmings (10 × 5 × 1 cm; approximately 100 g) fabricated from beef chuck rolls, and were immersed for 30 s in solutions of acidified sodium chlorite (0.1%, pH 2.5), peroxyacetic acid (0.02%, pH 3.8), sodium metasilicate (4%, pH 12.5), Bromitize® Plus (0.0225% active bromine, pH 6.6), or AFTEC 3000 (pH 1.2), or for 5 s in SYNTRx 3300 (pH 1.0). Each antimicrobial was tested independently together with an untreated control. Results showed that all tested decontamination treatments were similarly effective against the 6 nSTEC serogroups as they were against E. coli O157:H7. Irrespective of pathogen inoculum, treatment of beef trimmings with acidified sodium chlorite, peroxyacetic acid, or sodium metasilicate effectively (P < 0.05) reduced initial pathogen counts (3.4 to 3.9 log CFU/cm2) by 0.7 to 1.0, 0.6 to 1.0, and 1.3 to 1.5 log CFU/cm2, respectively. Reductions of pathogen counts (3.1 to 3.2 log CFU/cm2) by Bromitize Plus, AFTEC 3000, and SYNTRx 3300 were 0.1 to 0.4 log CFU/cm2, depending on treatment. Findings of this study should be useful to regulatory authorities and the meat industry as they consider nSTEC contamination in beef trimmings.

Practical Applications:  Findings of this study should be useful to: (i) meat processors as they design and conduct studies to validate the efficacy of antimicrobial treatments to control pathogen contamination on fresh beef products; and (ii) regulatory agencies as they consider approaches for better control of the studied pathogens.
Comparison of Variable‐Blade to Allo‐Kramer Shear Method in Assessing Rainbow Trout (Oncorhynchus mykiss) Fillet Firmness
Authors: Aussanasuwannakul Aunchalee , Slider Susan D. , Salem Mohamed , Yao Jianbo , Brett Kenney Patrick
Published: August 2012

Abstract:  A variable‐blade (VB) attachment was compared to the Allo‐Kramer (AK) shear attachment for texture analysis of rainbow trout fillets from 2 experiments; effects of attachment configuration, storage regimen, and cooking temperature are evaluated. In the 1st experiment, AK detected differences in force measurement, and VB showed that the perpendicular orientation yielded the highest response (P < 0.05). Fillets refrigerated (4 °C) for 0 d were firmer than fillets stored for 14 d (337.36 compared with 275.90 g/g). Raw fillets were firmer than cooked fillet (333.79 compared with 279.46 g/g). In the 2nd experiment, frozen storage at –25 °C for 30 d after refrigerated storage (R3F30 and R7F30) decreased VB shear force (P= 0.0019) and AK energy of shear (P= 0.0001) by 1.5‐ and 2‐fold compared to those evaluated after refrigerated storage for 3 and 7 d (R3 and R7), respectively. Cooking increased VB and AK texture for all storage regimens (P < 0.05). In both studies, instrumental texture did not correlate with alkaline‐insoluble hydroxyproline (P > 0.05). Shear direction affected force generated by the VB attachment, and this attachment could discriminate shear force differences due to cooking and frozen‐storage.

Practical Application:  Fillet texture was determined by a recently developed device and compared to texture determined by the Allo‐Kramer shear attachment; both responses were related to collagen content. The VB attachment defined fillet texture as affected by cooking and storage condition.
How to differentiate non‐erosive reflux disease from functional heartburn
Authors: Ke Mei Yun
Published: August 2012

Heartburn is a common symptom in gastroesophageal reflux disease. Endoscopic examination can differentiate between reflux esophagitis and non‐erosive reflux disease (NERD), but not between NERD and functional heartburn. With the development of new techniques, more NERD patients could be identified among those previously diagnosed with functional heartburn. Most patients with NERD, however, could be identified based on their clinical characteristics and response to proton pump inhibitors and/or integrated anti‐gastroesophageal reflux therapy.
Regulation of vascular endothelial growth factor by melatonin in human breast cancer cells
Authors: Alvarez‐García Virginia , González Alicia , Alonso‐González Carolina , Martínez‐Campa Carlos , Cos Samuel
Published: August 2012

Melatonin exerts oncostatic effects on breast cancer by interfering with the estrogen‐signaling pathways. Melatonin reduces estrogen biosynthesis in human breast cancer cells, surrounding fibroblasts and peritumoral endothelial cells by regulating cytokines that influence tumor microenvironment. This hormone also exerts antiangiogenic activity in tumoral tissue. In this work, our objective was to study the role of melatonin on the regulation of the vascular endothelial growth factor (VEGF) in breast cancer cells. To accomplish this, we cocultured human breast cancer cells (MCF‐7) with human umbilical vein endothelial cells (HUVECs). VEGF added to the cultures stimulated the proliferation of HUVECs and melatonin (1 mm) counteracted this effect. Melatonin reduced VEGF production and VEGF mRNA expression in MCF‐7 cells. MCF‐7 cells cocultured with HUVECs stimulated the endothelial cells proliferation and increased VEGF levels in the culture media. Melatonin counteracted both stimulatory effects on HUVECs proliferation and on VEGF protein levels in the coculture media. Conditioned media from MCF‐7 cells increased HUVECs proliferation, and this effect was significantly counteracted by anti‐VEGF and 1 mm melatonin. All these findings suggest that melatonin may play a role in the paracrine interactions between malignant epithelial cells and proximal endothelial cells through a downregulatory action on VEGF expression in human breast cancer cells, which decrease the levels of VEGF around endothelial cells. Lower levels of VEGF could be important in reducing the number of estrogen‐producing cells proximal to malignant cells as well as decreasing tumoral angiogenesis.
Therapeutic efficacy of melatonin in reducing retinal damage in an experimental model of early type 2 diabetes in rats
Authors: Salido Ezequiel M. , Bordone Melina , De Laurentiis Andrea , Chianelli Mónica , Keller Sarmiento María Inés , Dorfman Damián , Rosenstein Ruth E.
Published: August 2012

Diabetic retinopathy (DR) is a leading cause of acquired blindness in adults, mostly affected by type 2 diabetes mellitus (T2DM). We have developed an experimental model of early T2DM in adult rats which mimics some features of human T2DM at its initial stages and provokes significant retinal alterations. The aim of this work was to analyze the effect of melatonin on retinal changes induced by the moderate metabolic derangement. For this purpose, adult male Wistar rats received a control diet or 30% sucrose in the drinking water. Three weeks after this treatment, animals were injected with vehicle or streptozotocin (STZ, 25 mg/kg). One day or 3 wk after vehicle or STZ injection, animals were subcutaneously implanted with a pellet of melatonin. Fasting and postprandial glycemia, and glucose, and insulin tolerance tests were analyzed. At 12 wk of treatment, animals which received a sucrose‐enriched diet and STZ showed significant differences in metabolic tests, as compared with control groups. Melatonin, which did not affect glucose metabolism in control or diabetic rats, prevented the decrease in the electroretinogram a‐wave, b‐wave, and oscillatory potential amplitude, and the increase in retinal lipid peroxidation, NOS activity, TNFα, Müller cells glial fibrillary acidic protein, and vascular endothelial growth factor levels. In addition, melatonin prevented the decrease in retinal catalase activity. These results indicate that melatonin protected the retina from the alterations observed in an experimental model of DR associated with type 2 diabetes.
Incorporation of treatment plan spatial and temporal dose patterns into a prostate intrafractional motion management strategy
Authors: Zhang Pengpeng , Hunt Margie , Happersett Laura , Cox Brett , Mageras Gig
Published: August 2012

Purpose:

Periodic MV/KV radiographs taken during volumetric modulated arc therapy (VMAT) for hypofractionated treatment provide guidance in intrafractional motion management. The choice of imaging frequency and timing are key components in delivering the desired dose while reducing associated overhead such as imaging dose, preparation, and processing time. In this project the authors propose a paradigm with imaging timing and frequency based on the spatial and temporal dose patterns of the treatment plan.

Methods:

A number of control points are used in treatment planning to model VMAT delivery. For each control point, the sensitivity of individual target or organ‐at‐risk dose to motion can be calculated as the summation of dose degradations given the organ displacements along a number of possible motion directions. Instead of acquiring radiographs at uniform time intervals, MV/KV image pairs are acquired indexed to motion sensitivity. Five prostate patients treated via hypofractionated VMAT are included in this study. Intrafractional prostate motion traces from the database of an electromagnetic tracking system are used to retrospectively simulate the VMAT delivery and motion management. During VMAT delivery simulation patient position is corrected based on the radiographic findings via couch movement if target deviation violates a patient‐specific 3D threshold. The violation rate calculated as the percentage of traces failing the clinical dose objectives after motion correction is used to evaluate the efficacy of this approach.

Results:

Imaging indexed to a 10 s equitime interval and correcting patient position accordingly reduces the violation rate to 19.5% with intervention from 44.5% without intervention. Imaging indexed to the motion sensitivity further reduces the violation rate to 12.1% with the same number of images. To achieve the same 5% violation rate, the imaging incidence can be reduced by 40% by imaging indexed to motion sensitivity instead of time.

Conclusions:

The simulation results suggest that image scheduling according to the characteristics of the treatment plan can improve the efficiency of intrafractional motion management. Using such a technique, the accuracy of delivered dose during image‐guided hypofractionated VMAT treatment can be improved.
The Double-Edged Sword: Does Biomechanism Increase or Decrease Judges' Sentencing of Psychopaths?
Authors: Aspinwall Lisa G. , Brown Teneille R. , Tabery James
Published: August 2012

Biology Made Me Do It

Introducing evidence of a biological basis of dysfunctional behavior might induce a judge to impose a lighter sentence upon defendants on the grounds that their innate biology lessened personal responsibility for their actions. On the other hand, it might also plausibly lead to a longer sentence on the grounds that this biological predisposition would imply a higher risk of future criminal acts.

Aspinwall
et al.

(p.
846
) analyzed sentencing opinions from a nationwide group of judges in the United States of a scenario closely modeled on a real-life case. Varying two factors—whether evidence for a biological basis was presented and whether it was offered by the defense or by the prosecution—revealed an overall lessening of sentences when biology was invoked.
Long-Range Ordered Carbon Clusters: A Crystalline Material with Amorphous Building Blocks
Authors: Wang Lin , Liu Bingbing , Li Hui , Yang Wenge , Ding Yang , Sinogeikin Stanislav V. , Meng Yue , Liu Zhenxian , Zeng Xiao Cheng , Mao Wendy L.
Published: August 2012

Amorphous Crystals

One usually thinks of a crystal as containing atoms, molecules, or other ordered units arranged in a periodic fashion to make a larger structure.

L. Wang
et al.

(p.
825
; see the Perspective by

D. Wang and Fernandez-Martinez

) compressed a solvate of xylene in C
60
(fullerene) to very high pressures at room temperature, which caused the fullerene molecules to collapse and form amorphous clusters but with each cluster retaining its position in the original crystal lattice because of the interaction with the intercalated xylene molecules. Thus, a crystal was formed from amorphous constituents. On heating at atmospheric pressure, the xylene disappeared and the sample became completely amorphous.
Camouflage and Display for Soft Machines
Authors: Morin Stephen A. , Shepherd Robert F. , Kwok Sen Wai , Stokes Adam A. , Nemiroski Alex , Whitesides George M.
Published: August 2012

Mechanical Chameleon

A wide range of animals can adapt their color patterns as a means of camouflage or otherwise changing their appearance. This is accomplished through changes in coloration, contrast, patterning, or shape.

Morin
et al.

(p.
828
) show at a basic level that some of these features can be added as microfluidic layers attached to mobile, flexible, soft machines. By pumping different fluids through the channels, the robots were able to change their coloration or overall contrast and could thus blend into the background of the surface they were lying upon. Conversely, by pumping through fluids of different temperature, the infrared profile of the robot could be changed without changing its visible coloration.
Order from Disorder
Authors: Wang Dongbo , Fernandez-Martinez Alejandro
Published: August 2012

The repeat units of a crystalline material can be made up of disordered clusters.
Water Vapor in the Lower Stratosphere
Authors: Ravishankara A. R.
Published: August 2012

Water vapor injection into the stratosphere at mid-latitudes may sporadically enhance local stratospheric water vapor concentrations to high values.
Circadian Time Redoxed
Authors: Belle Mino D. C. , Piggins Hugh D.
Published: August 2012

Oscillations of the molecular clock and redox states control the electrical activity of mammalian clock neurons.
Policy Shifts on Emergency Use of Cholera Vaccines
Authors: Enserink Martin
Published: August 2012

This week, the World Health Organization is set to publish a report advocating for the creation of a global stockpile of cholera vaccines that would be rushed to countries when an outbreak begins.
Big or Small, Science Will Suffer If Sequestration Goes Into Effect
Authors: Mervis Jeffrey
Published: August 2012

On 2 January, $110 billion in budget cuts across all federal agencies are scheduled to take effect, which could be catastrophic for U.S. scientists doing research supported by the federal government.
Negative Report on GM Crops Shakes Government's Food Agenda
Authors: Bagla Pallava
Published: August 2012

A panel of the Indian Parliament last week recommended that GM crop "field trials under any garb should be discontinued forthwith," and that agricultural GM research should "only be done under strict containment."
Stopping Alzheimer's Before It Starts
Authors: Miller Greg
Published: August 2012

Three new clinical trials expected to begin next year will attempt to prevent dementia by treating people at risk for the disease before they develop symptoms.
How to Talk About Alzheimer's Risk
Authors: Miller Greg
Published: August 2012

Most people do not want to know if Alzheimer's disease is written in their genes, say researchers leading two trials that will try to prevent Alzheimer's in people with gene mutations that cause it.
As Isolation Ends, Myanmar Faces New Ecological Risks
Authors: Schmidt Charles
Published: August 2012

Concerned about the path of foreign investment, homegrown environmentalists seek to protect threatened forests and wildlife and push for sustainable development.
A Recombinant Clumping Factor A-Containing Vaccine Induces Functional Antibodies to Staphylococcus aureus That Are Not Observed after Natural Exposure
Authors: Hawkins Julio , Kodali Srinivas , Matsuka Yury V. , McNeil Lisa K. , Mininni Terri , Scully Ingrid L. , Vernachio John H. , Severina Elena , Girgenti Douglas , Jansen Kathrin U. , Anderson Annaliesa S. , Donald Robert G. K.
Published: August 2012

Staphylococcus aureusis a Gram-positive pathogen that causes devastating disease and whose pathogenesis is dependent on interactions with host cell factors. Staphylococcal clumping factor A (ClfA) is a highly conserved fibrinogen (Fg)-binding protein and virulence factor that contributes to host tissue adhesion and initiation of infection. ClfA is being investigated as a possible component of a staphylococcal vaccine. We report the development of an Fg-binding assay that is specific for ClfA-mediated binding. Using the assay, we show that despite the presence of anti-ClfA antibodies, human sera from unvaccinated subjects are unable to prevent the binding ofS. aureusto an Fg-coated surface. In contrast, antibodies elicited by a recombinant ClfA-containing vaccine were capable of blocking the ClfA-dependent binding of a diverse and clinically relevant collection of staphylococcal strains to Fg. These functional antibodies were also able to displaceS. aureusalready bound to Fg, suggesting that the ligand-binding activity of ClfA can be effectively neutralized through vaccination.
Gene Sequencing for Routine Verification of Pyrazinamide Resistance in Mycobacterium tuberculosis: a Role for
pncA
but Not
rpsA
Authors: Alexander David C. , Ma Jennifer H. , Guthrie Jennifer L. , Blair Joanne , Chedore Pam , Jamieson Frances B.
Published: August 2012

Pyrazinamide (PZA) is an important component of first-line therapy for the treatment of tuberculosis. Here, we evaluate targeted gene sequencing as a supplement to phenotypic PZA susceptibility testing of
Mycobacterium tuberculosis
. Routine sequencing of
pncA
, but not
rpsA
, is effective for verification of PZA susceptibility results.
Specific Detection of Enteroaggregative Hemorrhagic Escherichia coli O104:H4 Strains by Use of the CRISPR Locus as a Target for a Diagnostic Real-Time PCR
Authors: Delannoy Sabine , Beutin Lothar , Burgos Ylanna , Fach Patrick
Published: August 2012

In 2011, a large outbreak of an unusual bacterial strain occurred in Europe. This strain was characterized as a hybrid of an enteroaggregative
Escherichia coli
(EAEC) and a Shiga toxin-producing
E. coli
(STEC) strain of the serotype O104:H4. Here, we present a single PCR targeting the
c
lustered
r
egularly
i
nterspaced
s
hort
p
alindromic
r
epeats locus of
E. coli
O104:H4 (CRISPR
O104:H4
) for specific detection of EAEC STEC O104:H4 strains from different geographical locations and time periods. The specificity of the CRISPR
O104:H4
PCR was investigated using 1,321
E. coli
strains, including reference strains for
E. coli
O serogroups O1 to O186 and flagellar (H) types H1 to H56. The assay was compared for specificity using PCR assays targeting different O104 antigen-encoding genes (
wbwC
O104
,
wzx
O104
, and
wzy
O104
). The PCR assays reacted with all types of
E. coli
O104 strains (O104:H2, O104:H4, O104:H7, and O104:H21) and with
E. coli
O8 and O9 strains carrying the K9 capsular antigen and were therefore not specific for detection of the EAEC STEC O104:H4 type. A single PCR developed for the CRISPR
O104:H4
target was sufficient for specific identification and detection of the 48 tested EAEC STEC O104:H4 strains. The 35
E. coli
O104 strains expressing H types other than H4 as well as 8
E. coli
strains carrying a K9 capsular antigen tested all negative for the CRISPR
O104:H4
locus. Only 12 (0.94%) of the 1,273 non-O104:H4
E. coli
strains (serotypes Ont:H2, O43:H2, O141:H2, and O174:H2) reacted positive in the CRISPR
O104:H4
PCR (99.06% specificity).
Development of a TaqMan Multiplex PCR Assay for Detection of Plasmid-Mediated AmpC β-Lactamase Genes
Authors: Geyer Chelsie N. , Reisbig Mark D. , Hanson Nancy D.
Published: August 2012

A multiplex, real-time TaqMan assay was designed to identify clinical isolates carrying plasmid-mediatedampCgenes. The specificity and sensitivity of this assay were 100% when testing characterized AmpC/non-AmpC-producing isolates and randomly selected clinical isolates. This is a rapid assay that can be performed in a clinical microbiology laboratory.
Evaluation of the Sensititre MycoTB Plate for Susceptibility Testing of the Mycobacterium tuberculosis Complex against First- and Second-Line Agents
Authors: Hall Leslie , Jude Kurt P. , Clark Shirley L. , Dionne Kim , Merson Ryan , Boyer Ana , Parrish Nicole M. , Wengenack Nancy L.
Published: August 2012

The Sensititre MycoTB plate (TREK Diagnostic Systems, Cleveland, OH) uses a microtiter plate MIC format for susceptibility testing of
Mycobacterium tuberculosis
complex isolates against first- and second-line antituberculosis agents. Categorical agreement versus the agar proportion method for 122
M. tuberculosis
complex isolates was 94% to 100%.
Clonality and Antimicrobial Susceptibility of Staphylococcus aureus and Methicillin-Resistant S. aureus Isolates from Food Animals and Other Animals
Authors: Ho Pak-Leung , Chow Kin-Hung , Lai Eileen L. , Law Pierra Y. T. , Chan Pui-Ying , Ho Alex Y. M. , Ng Tak-Keung , Yam Wing-Cheong
Published: August 2012

Out of 3,081 animals studied, 24.9% of pigs, 4.7% of chickens, 6.3% of dogs, 10.5% of cats, and 7.1% of rodents were
Staphylococcus aureus
positive. Prevalence of methicillin-resistant
S. aureus
(MRSA) was high in pigs (animals, 21.3%; batches, 46.5%), with all MRSA isolates and most methicillin-sensitive
S. aureus
isolates belonging to clonal complex 9 (CC9) and being multidrug resistant. The predominant
S. aureus
CCs among dog and cat isolates were similar. Among rodent isolates, CC398 predominated, with
spa
t034 the most frequent
spa
type detected.
Protective Vaccine-Induced CD4+T Cell-Independent B Cell Responses against Rabies Infection
Authors: Dorfmeier Corin L. , Lytle Andrew G. , Dunkel Amber L. , Gatt Anthony , McGettigan James P.
Published: August 2012

A major goal in rabies virus (RV) research is to develop a single-dose postexposure prophylaxis (PEP) that would simplify vaccination protocols, reduce costs associated with rabies prevention in humans, and save lives. Live replication-deficient RV-based vaccines are emerging as promising single-dose vaccines to replace currently licensed inactivated RV-based vaccines. Nonetheless, little is known about how effective B cells develop in response to live RV-based vaccination. Understanding this fundamental property of rabies immunology may help in developing a single-dose RV vaccine. Typically, vaccines induce B cells secreting high-affinity, class-switched antibodies during germinal center (GC) reactions; however, there is a lag time between vaccination and the generation of GC B cells. In this report, we show that RV-specific antibodies are detected in mice immunized with live but not inactivated RV-based vaccines before B cells displaying a GC B cell phenotype (B220+GL7hiCD95hi) are formed, indicating a potential role for T cell-independent and early extrafollicular T cell-dependent antibody responses in the protection against RV infection. Using two mouse models of CD4+T cell deficiency, we show that B cells secreting virus-neutralizing antibodies (VNAs) are induced via T cell-independent mechanisms within 4 days postimmunization with a replication-deficient RV-based vaccine. Importantly, mice that are completely devoid of T cells (B6.129P2-Tcrβtm1MomTcrδtm1Mom/J) show protection against pathogenic challenge shortly after immunization with a live replication-deficient RV-based vaccine. We show that vaccines that can exploit early pathways of B cell activation and development may hold the key for the development of a single-dose RV vaccine wherein the rapid induction of VNA is critical.
U20 Is Responsible for Human Herpesvirus 6B Inhibition of Tumor Necrosis Factor Receptor-Dependent Signaling and Apoptosis
Authors: Kofod-Olsen Emil , Ross-Hansen Katrine , Schleimann Mariane Høgsbjerg , Jensen Dea Kejlberg , Møller Janni Michelle Lund , Bundgaard Bettina , Mikkelsen Jacob Giehm , Höllsberg Per
Published: August 2012

The immune system targets virus-infected cells by different means. One of the essential antiviral mechanisms is apoptosis induced by ligation of tumor necrosis factor receptor 1 (TNFR1). This receptor can be activated by tumor necrosis factor alpha (TNF-α), which upon binding to TNFR1 induces the assembly of first an inflammatory and later a proapoptotic signaling complex. Here, we report that infection by human herpesvirus 6B (HHV-6B) inhibited poly(ADP-ribose) polymerase (PARP) cleavage, caspase 3 and 8 activation, and IκBα Ser-32 phosphorylation downstream of TNFR1, indicating inhibition of both the inflammatory and apoptotic signaling pathways. We identified a hitherto uncharacterized viral protein, U20, as sufficient for mediating this inhibition. U20 was shown to locate to the cell membrane, and overexpression inhibited PARP cleavage, caspase 3 and 8 activation, IκBα Ser-32 phosphorylation, and NF-κB transcriptional activity. Moreover, small interfering RNA (siRNA) knockdown of U20 demonstrated that the protein is necessary for HHV-6B-mediated inhibition of TNFR signaling during infection. These results suggest an important novel function of U20 as a viral immune evasion protein during HHV-6B infection.
Koi Herpesvirus Encodes and Expresses a Functional Interleukin-10
Authors: Sunarto Agus , Liongue Clifford , McColl Kenneth A. , Adams Mathew M. , Bulach Dieter , Crane Mark St. J. , Schat Karel A. , Slobedman Barry , Barnes Andrew C. , Ward Alister C. , Walker Peter J.
Published: August 2012

Koi herpesvirus (KHV) (speciesCyprinid herpesvirus 3) ORF134 was shown to transcribe a spliced transcript encoding a 179-amino-acid (aa) interleukin-10 (IL-10) homolog (khvIL-10) in koi fin (KF-1) cells. Pairwise sequence alignment indicated that the expressed product shares 25% identity with carp IL-10, 22 to 24% identity with mammalian (including primate) IL-10s, and 19.1% identity with European eel herpesvirus IL-10 (ahvIL-10). In phylogenetic analyses, khvIL-10 fell in a divergent position from all host IL-10 sequences, indicating extensive structural divergence following capture from the host. In KHV-infected fish, khvIL-10 transcripts were observed to be highly expressed during the acute and reactivation phases but to be expressed at very low levels during low-temperature-induced persistence. Similarly, KHV early (helicase [Hel] and DNA polymerase [DNAP]) and late (intercapsomeric triplex protein [ITP] and major capsid protein [MCP]) genes were also expressed at high levels during the acute and reactivation phases, but only low-level expression of the ITP gene was detected during the persistent phase. Injection of khvIL-10 mRNA into zebrafish (Danio rerio) embryos increased the number of lysozyme-positive cells to a similar degree as zebrafish IL-10. Downregulation of the IL-10 receptor long chain (IL-10R1) using a specific morpholino abrogated the response to both khvIL-10 and zebrafish IL-10 transcripts, indicating that, despite the structural divergence, khvIL-10 functions via this receptor. This is the first report describing the characteristics of a functional viral IL-10 gene in theAlloherpesviridae.
Reconstitution of the Entire Hepatitis C Virus Life Cycle in Nonhepatic Cells
Authors: Da Costa Daniel , Turek Marine , Felmlee Daniel J. , Girardi Erika , Pfeffer Sébastien , Long Gang , Bartenschlager Ralf , Zeisel Mirjam B. , Baumert Thomas F.
Published: August 2012

Hepatitis C virus (HCV) is a human hepatotropic virus, but the relevant host factors restricting HCV infection to hepatocytes are only partially understood. We demonstrate that exogenous expression of defined host factors reconstituted the entire HCV life cycle in human nonhepatic 293T cells. This study shows robust HCV entry, RNA replication, and production of infectious virus in human nonhepatic cells and highlights key host factors required for liver tropism of HCV.
Sheeppox Virus SPPV14 Encodes a Bcl-2-Like Cell Death Inhibitor That Counters a Distinct Set of Mammalian Proapoptotic Proteins
Authors: Okamoto Toru , Campbell Stephanie , Mehta Ninad , Thibault John , Colman Peter M. , Barry Michele , Huang David C. S. , Kvansakul Marc
Published: August 2012

Many viruses express inhibitors of programmed cell death (apoptosis), thereby countering host defenses that would otherwise rapidly clear infected cells. To counter this, viruses such as adenoviruses and herpesviruses express recognizable homologs of the mammalian prosurvival protein Bcl-2. In contrast, the majority of poxviruses lack viral Bcl-2 (vBcl-2) homologs that are readily identified by sequence similarities. One such virus, myxoma virus, which is the causative agent of myxomatosis, expresses a virulence factor that is a potent inhibitor of apoptosis. In spite of the scant sequence similarity to Bcl-2, myxoma virus M11L adopts an almost identical 3-dimensional fold. We used M11L as bait in a sequence similarity search for other Bcl-2-like proteins and identified six putative vBcl-2 proteins from poxviruses. Some are potent inhibitors of apoptosis, in particular sheeppox virus SPPV14, which inhibited cell death induced by multiple agents. Importantly, SPPV14 compensated for the loss of antiapoptotic F1L in vaccinia virus and acts to directly counter the cell death mediators Bax and Bak. SPPV14 also engages a unique subset of the death-promoting BH3-only ligands, including Bim, Puma, Bmf, and Hrk. This suggests that SPPV14 may have been selected for specific biological roles as a virulence factor for sheeppox virus.
Epstein-Barr Virus Transcription Activator R Upregulates BARF1 Expression by Direct Binding to Its Promoter, Independent of Methylation
Authors: Hoebe E. K. , Wille C. , Hopmans E. S. , Robinson A. R. , Middeldorp J. M. , Kenney S. C. , Greijer A. E.
Published: August 2012

Epstein-Barr virus (EBV) BamHI-A rightward frame 1 (BARF1) is considered a major viral oncogene in epithelial cells and has immune-modulating properties. However, in B cells and lymphomas, BARF1 expression is restricted to the viral lytic replication cycle. In this report, the transcriptional regulation of BARF1 during lytic replication is unraveled. Bisulfite sequencing of various cell lines indicated a high level of methylation of the BARF1 gene control region. A BARF1 promoter luciferase reporter construct was created using a CpG-free vector, enabling true assessment of promoter methylation. Induction of the EBV lytic cycle is mediated by the immediate-early proteins BZLF1 (Z) and BRLF1 (R). R was found to activate expression of the BARF1 promoter up to 250-fold independently of Z and unaffected by BARF1 promoter methylation. Chromatin immunoprecipitation (ChIP), electrophoretic mobility shift assay (EMSA), and specific mutagenesis of the R-responsive elements (RREs) demonstrated direct binding of R to RREs between nucleotides −554 and −327 relative to the BARF1 transcriptional ATG start site. The kinetics of BARF1 expression upon transactivation by R showed that BARF1 mRNA was expressed within 6 h in the context of the viral genome. In conclusion, expression of the BARF1 protein during lytic replication is regulated by direct binding of R to multiple RREs in the gene control region and is independent of the promoter methylation status. The early kinetics of BARF1 upon transactivation by R confirm its status as an early gene and emphasize the necessity of early immune modulation during lytic reactivation.
Oral Clarithromycin Enhances Airway Immunoglobulin A (IgA) Immunity through Induction of IgA Class Switching Recombination and B-Cell-Activating Factor of the Tumor Necrosis Factor Family Molecule on Mucosal Dendritic Cells in Mice Infected with Influenza A Virus
Authors: Takahashi Etsuhisa , Kataoka Kosuke , Indalao Irene L. , Konoha Keiko , Fujii Kazuyuki , Chida Junji , Mizuno Dai , Fujihashi Kohtaro , Kido Hiroshi
Published: August 2012

We previously reported that the macrolide antibiotic clarithromycin (CAM) enhanced the mucosal immune response in pediatric influenza, particularly in children treated with the antiviral neuraminidase inhibitor oseltamivir (OSV) with low production of mucosal antiviral secretory IgA (S-IgA). The aims of the present study were to confirm the effects of CAM on S-IgA immune responses, by using influenza A virus (IAV) H1N1-infected mice treated with or without OSV, and to determine the molecular mechanisms responsible for the induction of mucosal IgA class switching recombination in IAV-infected CAM-treated mice. The anti-IAV S-IgA responses and expression levels of IgA class switching recombination-associated molecules were examined in bronchus-lymphoid tissues and spleens of infected mice. We also assessed neutralization activities of S-IgA against IAV. Data show that CAM enhanced anti-IAV S-IgA induction in the airway of infected mice and restored the attenuated antiviral S-IgA levels in OSV-treated mice to the levels in the vehicle-treated mice. The expression levels of B-cell-activating factor of the tumor necrosis factor family (BAFF) molecule on mucosal dendritic cells as well as those of activation-induced cytidine deaminase and Iμ-Cα transcripts on B cells were enhanced by CAM, compared with the levels without CAM treatment, but CAM had no effect on the expression of the BAFF receptor on B cells. Enhancement by CAM of neutralization activities of airway S-IgA against IAV
in vitro
and reinfected mice was observed. This study identifies that CAM enhances S-IgA production and neutralizing activities through the induction of IgA class switching recombination and upregulation of BAFF molecules in mucosal dendritic cells in IAV-infected mice.
Evaluation of Phosphatidylinositol-4-Kinase IIIα as a Hepatitis C Virus Drug Target
Authors: Vaillancourt Frédéric H. , Brault Martine , Pilote Louise , Uyttersprot Nathalie , Gaillard Elias T. , Stoltz James H. , Knight Brian L. , Pantages Lynn , McFarland Mary , Breitfelder Steffen , Chiu Tim T. , Mahrouche Louiza , Faucher Anne-Marie , Cartier Mireille , Cordingley Michael G. , Bethell Richard C. , Jiang Huiping , White Peter W. , Kukolj George
Published: August 2012

Phosphatidylinositol-4-kinase IIIα (PI4KIIIα) is an essential host cell factor for hepatitis C virus (HCV) replication. An N-terminally truncated 130-kDa form was used to reconstitute an
in vitro
biochemical lipid kinase assay that was optimized for small-molecule compound screening and identified potent and specific inhibitors. Cell culture studies with PI4KIIIα inhibitors demonstrated that the kinase activity was essential for HCV RNA replication. Two PI4KIIIα inhibitors were used to select cell lines harboring HCV replicon mutants with a 20-fold loss in sensitivity to the compounds. Reverse genetic mapping isolated an NS4B-NS5A segment that rescued HCV RNA replication in PIK4IIIα-deficient cells. HCV RNA replication occurs on specialized membranous webs, and this study with PIK4IIIα inhibitor-resistant mutants provides a genetic link between NS4B/NS5A functions and PI4-phosphate lipid metabolism. A comprehensive assessment of PI4KIIIα as a drug target included its evaluation for pharmacologic intervention
in vivo
through conditional transgenic murine lines that mimic target-specific inhibition in adult mice. Homozygotes that induce a knockout of the kinase domain or knock in a single amino acid substitution, kinase-defective PI4KIIIα, displayed a lethal phenotype with a fairly widespread mucosal epithelial degeneration of the gastrointestinal tract. This essential host physiologic role raises doubt about the pursuit of PI4KIIIα inhibitors for treatment of chronic HCV infection.