bluuebunny/crossref_metadata_2025 huggingface dataset:
New Public Management Reforms and Efficiency in Urban Water Service Delivery in Developing Countries
Authors: Mukokoma Mary Maurice Nalwoga , van Dijk Meine Pieter
Published: September 2012

Managing publicly owned entities using private sector principles, commonly known as New Public Management (NPM), is a widespread reform in Developing Countries. The theoretical and empirical debate on the effect of NPM on the performance of Public entities is still ongoing, yet limited research has been conducted in the water sector in developing countries. Using Data Envelopment Analysis (DEA) and Tobit regression on 300 observations, the influence of adopting NPM reform dimensions of segregation of functions, managerial autonomy, accountability for results, customer orientation, and market orientation on technical efficiency of urban public water utilities has been addressed. The results indicate a high likelihood of the NPM reform causing a change in the technical efficiency of the Decision-Making Units (DMUs) in National Water and Sewerage Corporation (NWSC) in Uganda though it was not the case for the DMUs of Dar es Salaam Water and Sewerage Corporation (DAWASCO) in Tanzania.
The Time Has Come
Authors: Kruschke John K. , Aguinis Herman , Joo Harry
Published: September 2012

The use of Bayesian methods for data analysis is creating a revolution in fields ranging from genetics to marketing. Yet, results of our literature review, including more than 10,000 articles published in 15 journals from January 2001 and December 2010, indicate that Bayesian approaches are essentially absent from the organizational sciences. Our article introduces organizational science researchers to Bayesian methods and describes why and how they should be used. We use multiple linear regression as the framework to offer a step-by-step demonstration, including the use of software, regarding how to implement Bayesian methods. We explain and illustrate how to determine the prior distribution, compute the posterior distribution, possibly accept the null value, and produce a write-up describing the entire Bayesian process, including graphs, results, and their interpretation. We also offer a summary of the advantages of using Bayesian analysis and examples of how specific published research based on frequentist analysis-based approaches failed to benefit from the advantages offered by a Bayesian approach and how using Bayesian analyses would have led to richer and, in some cases, different substantive conclusions. We hope that our article will serve as a catalyst for the adoption of Bayesian methods in organizational science research.
Introducing Functional Data Analysis to Managerial Science
Authors: Dass Mayukh , Shropshire Christine
Published: September 2012

In this article, we introduce functional data analysis (FDA), a set of statistical tools developed to study information on curves or functions. We review fundamentals of the methodology along with previous applications in other business disciplines to highlight the potential of FDA to managerial science. We provide details of the three most commonly used FDA techniques, including functional principal component analysis, functional regression, and functional clustering, and demonstrate each by investigating measures of firm financial performance from a panel data set of the 1,000 largest U.S. firms by revenues from 1992 to 2008. We compare results obtained from FDA with hierarchical linear modeling and conclude by outlining ideas for future micro- and macro-level organizational research incorporating this methodology.
Bone Ultrasound and Physical Activity in Postmenopausal Spanish Women
Authors: Calderon-Garcia Julian F. , Lavado-Garcia Jesus M. , Martin Raul Roncero , Moran Jose M. , Canal-Macias Maria L. , Pedrera-Zamorano Juan D.
Published: September 2012

We studied the effects of moderate physical activity on bone mass in healthy postmenopausal women as assessed by quantitative bone ultrasound. We enrolled a total of 326 postmenopausal women (60.95 ± 8.51 years old and 13.02 ± 9.52 years since menopause) and categorized them by weekly physical activity in their leisure time (sedentary, moderate, and active). We administered a questionnaire on general health and diet and measured bone density by ultrasound on phalanges II–V in the nondominant hand. We found no significant difference in the amplitude-dependent speed of sound (Ad-SoS; p > .05) between sedentary and active women. We concluded that the changes induced by moderate physical activity on bone mass are minimal in healthy postmenopausal women.
Assessing Spoken Language Competence in Children With Selective Mutism
Authors: Klein Evelyn R. , Armstrong Sharon Lee , Shipon-Blum Elisa
Published: September 2012

Children with selective mutism (SM) display a failure to speak in select situations despite speaking when comfortable. The purpose of this study was to obtain valid assessments of receptive and expressive language in 33 children (ages 5 to 12) with SM. Because some children with SM will speak to parents but not a professional, another purpose was to explore the efficacy of employing parents to deliver test stimuli. Parents received training on presenting standardized test material and were monitored during testing by a professional who scored and interpreted the results. Professional-administered tests underestimated children’s capabilities. However, even with parents, children’s scores decreased as the tasks changed from receptive to expressive vocabulary and from narrative comprehension to telling a story on their own. Thus, although SM is typically classified as an anxiety disorder, an underlying expressive narrative language deficit was identified in 42% of children with SM using this new procedure.
Exploring Interagency Collaboration in AAC Intervention
Authors: Alant Erna , Champion Annette , Peabody Erin Colone
Published: September 2012

This study grew from a need identified by professionals working in the same community to explore interagency support for augmentative and alternative communication device implementation with students, families, and professionals involving the local school system and university. A case study was used to document intervention with a second-grade student who had developmental and severe expressive language delays. Based on the collaboration that was initiated between university and school corporation during the device acquisition process, it was determined that an interagency support system for this student would provide the best long-term solution for training, implementation, and maintenance related to her device. The participant’s intervention progress before and after intervention was described using tests and video interactions. Interviews with the principal collaborators (two speech and language pathologists) and the parent were conducted post intervention. Findings revealed positive changes in performance on the receptive and expressive vocabulary testing and scores on communication functions used based on video analyses before and after intervention. Spontaneous device use at home and school was, however, limited. As the study represented one case, no further statistical analysis was conducted. Finally, some reflections on the process were provided by the two main interagency collaborators as well as the mother of the participant, and implications for future intervention were discussed.
Barriers to Retention in Care as Perceived by Persons Living with HIV in Rural Ethiopia
Authors: Lifson Alan R. , Demissie Workneh , Tadesse Alemayehu , Ketema Kassu , May Randy , Yakob Bereket , Metekia Meka , Slater Lucy , Shenie Tibebe
Published: September 2012

Inability to retain HIV-infected patients in care undermines the benefits of starting millions in low-income countries on antiretroviral therapy (ART). In a hospital HIV clinic in rural southern Ethiopia, we conducted focus groups of HIV-infected men and women to learn more about experiences with and barriers to attending clinic appointments. Respondents reported multiple barriers, including those that were patient related (eg, misunderstandings about ART, mistaken belief in AIDS cures, and drug/alcohol use), clinic related (eg, negative provider interactions, lack of familiarity with patients’ medical situation, and overcrowding), medication related (eg, side effects), social (eg, stigma and discrimination and lack of support), and situational/resource related (eg, distance to clinic, lack of funds, competing domestic/work priorities, and lack of food). Based on the lessons learned from these focus groups, we implemented a community intervention to improve retention, using trained community support workers who provide patient education, counseling, social support, problem-solving assistance, needed referrals, and improved communication/linkage to the patients’ HIV clinic.
Perceived Family Support and Depression among People Living with HIV/AIDS in the Kathmandu Valley, Nepal
Authors: Shrestha Sadhana , Poudel Krishna C. , Poudel-Tandukar Kalpana , Kobayashi Jun , Pandey Basu Dev , Yasuoka Junko , Otsuka Keiko , Jimba Masamine
Published: September 2012

Background:

Depression is emerging as a highly prevalent psychiatric condition among people living with HIV/AIDS (PLWHA). Perceived family support (PFS) buffers depression among chronic disease patients. However, a similar relationship among PLWHA is unexplored.

Objective:

To examine the relationship between PFS and depression among PLWHA in the Kathmandu Valley, Nepal.

Methods:

In this cross-sectional study, depression was measured by Beck Depression Inventory Ia. Perceived family support was measured by Nepali Family Support and Difficulties Scale. The status of depression was compared between 208 PLWHA and 208 HIV-negative participants. The relationship between PFS and depression was examined only among PLWHA.

Results:

Among each of the 208 participants, the number of depressed PLWHA (n = 61,29.3%) was higher than that of HIV-negative participants (n = 13,6.2%; P < .001). Perceived family support had a negative association with depression in PLWHA (adjusted odds ratio [AOR] = 0.20, 95% confidence interval [CI] = 0.08-0.53).

Conclusion:

In Nepal, PLWHA display a higher level of depression than HIV-negative people, and a lower level of PFS is associated with depression among PLWHA. Improved family support might be helpful in reducing depression among Nepalese PLWHA.
Longitudinal Evaluation of Resting-State fMRI After Acute Stroke With Hemiparesis
Authors: Golestani Ali-Mohammad , Tymchuk Sarah , Demchuk Andrew , Goodyear Bradley G. ,   VISION-2 Study Group
Published: September 2012

Background. Functional magnetic resonance imaging (fMRI) of motor impairment after stroke strongly depends on patient effort and capacity to make a movement. Hence fMRI has had limited use in clinical management. Alternatively, resting-state fMRI (ie, with no task) can elucidate the brain’s functional connections by determining temporal synchrony between brain regions. Objective. The authors examined whether resting-state fMRI can elucidate the disruption of functional connections within hours of ischemic stroke as well as during recovery. Methods. A total of 51 ischemic stroke patients—31 with mild-to-moderate hand deficits (National Institutes of Health Stroke Scale [NIHSS] motor score ≥1) and 20 with NIHSS score of 0—underwent resting-state fMRI at <24 hours, 7 days, and 90 days poststroke; 15 age-matched healthy individuals participated in 1 session. Using the resting-state fMRI signal from the ipsilesional motor cortex, the strength of functional connections with the contralesional motor cortex was computed. Whole-brain maps of the resting-state motor network were also generated and compared between groups and sessions. Results. Within hours poststroke, patients with motor deficits exhibited significantly lower connectivity than controls ( P = .02) and patients with no motor impairment ( P = .03). Connectivity was reestablished after 7 days in recovered (ie, NIHSS score = 0) participants. After 90 days, recovered patients exhibited normal motor connectivity; however, reduced connectivity with subcortical regions associated with effort and cognitive processing remained. Conclusion. Resting-state fMRI within hours of ischemic stroke can demonstrate the impact of stroke on functional connections throughout the brain. This tool has the potential to help select appropriate stroke therapies in an acute imaging setting and to monitor the efficacy of rehabilitation.
Profile of an Effective Urban Music Educator
Authors: Baker Vicki D.
Published: September 2012

The purpose of this study was to develop a profile of an effective urban music educator in an effort to provide strategies for university teacher training programs to prepare students to teach in urban schools. The study examined urban music teachers’ ( N = 158) educational background, effective and ineffective characteristics, perceived challenges, and traits needed to sustain a long-term career. Results indicated that participants who attended urban schools themselves had longer tenures in urban schools than those who had not. The greatest challenges facing urban educators included discipline, uninvolved/unsupportive parents, cultural diversity, students’ limited musical training/exposure, and unstable home environments. An effective urban music educator was described as being empathetic, patient, flexible, enthusiastic, and committed to students’ learning. Career urban music teachers were portrayed as being flexible, compassionate/caring, determined/persistent, committed to working hard, loving and respectful of students, patient, and knowledgeable. Results are discussed in terms of implications for teacher preparation.
Water-based exercise in COPD with physical comorbidities: a randomised controlled trial
Authors: McNamara Renae J. , McKeough Zoe J. , McKenzie David K. , Alison Jennifer A.
Published: September 2012

Land-based exercise is often difficult for people with chronic obstructive pulmonary disease (COPD) who have coexisting obesity or musculoskeletal or neurological conditions. This randomised controlled trial aimed to determine the effectiveness of water-based exercise training in improving exercise capacity and quality of life compared to land-based exercise training and control (no exercise) in people with COPD and physical comorbidities.

Participants referred to pulmonary rehabilitation were randomly allocated to a water-based exercise, land-based exercise or the control group. The two exercise groups trained for 8 weeks, completing three sessions per week.

45 out of 53 participants (mean±sdage 72±9 years; forced expiratory volume in 1 s 59±15% predicted) completed the study. Compared to controls, water-based exercise training significantly increased 6-min walking distance, incremental and endurance shuttle walk distances, and improved Chronic Respiratory Disease Questionnaire (CRDQ) dyspnoea and fatigue. Compared to land-based exercise training, water-based exercise training significantly increased incremental shuttle walk distance (mean difference 39 m, 95% CI 5–72 m), endurance shuttle walk distance (mean difference 228 m, 95% CI 19–438 m) and improved CRDQ fatigue.

Water-based exercise training was significantly more effective than land-based exercise training and control in increasing peak and endurance exercise capacity and improving aspects of quality of life in people with COPD and physical comorbidities.
Efficacy and safety of meropenem–clavulanate added to linezolid-containing regimens in the treatment of MDR-/XDR-TB
Authors: De Lorenzo Saverio , Alffenaar Jan Wilem , Sotgiu Giovanni , Centis Rosella , D'Ambrosio Lia , Tiberi Simon , Bolhuis Mathieu S. , van Altena Richard , Viggiani Piero , Piana Andrea , Spanevello Antonio , Migliori Giovanni Battista
Published: September 2012

Clinical experience on meropenem–clavulanate to treat tuberculosis (TB) is anecdotal (according to case reports on 10 patients). The aim of our case–control study was to evaluate the contribution of meropenem–clavulanate when added to linezolid-containing regimens in terms of efficacy and safety/tolerability in treating multidrug-resistant (MDR) and extensively drug-resistant (XDR) TB cases after 3 months of second-line treatment.

37 cases with MDR-/XDR-TB were prescribed meropenem–clavulanate (3 g daily dose) in addition to a linezolid-containing regimen (dosage range 300–1200 mg·day−1), designed according to international guidelines, which was prescribed to 61 controls.

The clinical severity of cases was worse than that of controls (drug susceptibility profile, proportion of sputum-smear positive and of re-treatment cases). The group of cases yielded a higher proportion of sputum-smear converters (28 (87.5%) out of 32versusnine (56.3%) out of 16; p=0.02) and culture converters (31 (83.8%) out of 37versus15 (62.5%) out of 24; p=0.06). Excluding XDR-TB patients (11 (11.2%) out of 98), cases scored a significantly higher proportion of culture converters than controls (p=0.03). One case had to withdraw from meropenem–clavulanate due to increased transaminase levels.

The results of our study provide: 1) preliminary evidence on effectiveness and safety/tolerability of meropenem–clavulanate; 2) reference to design further trials; and 3) a guide to clinicians for its rationale use within salvage/compassionate regimens.
Pulmonary arteriole gene expression signature in idiopathic pulmonary fibrosis
Authors: Patel Nina M. , Kawut Steven M. , Jelic Sanja , Arcasoy Selim M. , Lederer David J. , Borczuk Alain C.
Published: September 2012

A third of patients with idiopathic pulmonary fibrosis (IPF) develop pulmonary hypertension (PH-IPF), which is associated with increased mortality. Whether an altered gene expression profile in the pulmonary vasculature precedes the clinical onset of PH-IPF is unknown. We compared gene expression in the pulmonary vasculature of IPF patients with and without PH with controls.

Pulmonary arterioles were isolated using laser capture microdissection from 16 IPF patients: eight with PH (PH-IPF) and eight with no PH (NPH-IPF), and seven controls. Probe was prepared from extracted RNA, and hybridised to Affymetrix Hu133 2.0 Plus genechips. Biometric Research Branch array tools and Ingenuity Pathway Analysis software were used for analysis of the microarray data.

Univariate analysis revealed 255 genes that distinguished IPF arterioles from controls (p<0.001). Mediators of vascular smooth muscle and endothelial cell proliferation, Wnt signalling and apoptosis were differentially expressed in IPF arterioles. Unsupervised and supervised clustering analyses revealed similar gene expression in PH-IPF and NPH-IPF arterioles.

The pulmonary arteriolar gene expression profile is similar in IPF patients with and without coexistent PH. Pathways involved in vascular proliferation and aberrant apoptosis, which may contribute to pulmonary vascular remodelling, are activated in IPF patients.
Analysis of TIR- and non-TIR-NBS-LRR disease resistance gene analogous in pepper: characterization, genetic variation, functional divergence and expression patterns
Authors: Wan Hongjian , Yuan Wei , Ye Qingjing , Wang Rongqing , Ruan Meiying , Li Zhimiao , Zhou Guozhi , Yao Zhuping , Zhao Jing , Liu Shujun , Yang Yuejian
Published: September 2012

Background

Pepper (Capsicum annuum L.) is one of the most important vegetable crops worldwide. However, its yield and fruit quality can be severely threatened by several pathogens. The plant nucleotide-binding site (NBS)-leucine-rich repeat (LRR) gene family is the largest class of known disease resistance genes (R genes) effective against such pathogens. Therefore, the isolation and identification of such R gene homologues from pepper will provide a critical foundation for improving disease resistance breeding programs.

Results

A total of 78 R gene analogues (CaRGAs) were identified in pepper by degenerate PCR amplification and database mining. Phylogenetic tree analysis of the deduced amino acid sequences for 51 of these CaRGAs with typically conserved motifs ( P-loop, kinase-2 and GLPL) along with some known R genes from Arabidopsis and tomato grouped these CaRGAs into the non-Toll interleukin-1 receptor (TIR)-NBS-LRR (CaRGAs I to IV) and TIR-NBS-LRR (CaRGAs V to VII) subfamilies. The presence of consensus motifs (i.e. P-loop, kinase-2 and hydrophobic domain) is typical of the non-TIR- and TIR-NBS-LRR gene subfamilies. This finding further supports the view that both subfamilies are widely distributed in dicot species. Functional divergence analysis provided strong statistical evidence of altered selective constraints during protein evolution between the two subfamilies. Thirteen critical amino acid sites involved in this divergence were also identified using DIVERGE version 2 software. Analyses of non-synonymous and synonymous substitutions per site showed that purifying selection can play a critical role in the evolutionary processes of non-TIR- and TIR-NBS-LRR RGAs in pepper. In addition, four specificity-determining positions were predicted to be responsible for functional specificity. qRT-PCR analysis showed that both salicylic and abscisic acids induce the expression of CaRGA genes, suggesting that they may primarily be involved in defence responses by activating signaling pathways.

Conclusion

The identified CaRGAs are a valuable resource for discovering R genes and developing RGA molecular markers for genetic map construction. They will also be useful for improving disease resistance in pepper. The findings of this study provide a better understanding of the evolutionary mechanisms that drive the functional diversification of non-TIR- and TIR-NBS-LRR R genes in pepper.
Proteomic analysis of Clostridium thermocellum core metabolism: relative protein expression profiles and growth phase-dependent changes in protein expression
Authors: Rydzak Thomas , McQueen Peter D , Krokhin Oleg V , Spicer Vic , Ezzati Peyman , Dwivedi Ravi C , Shamshurin Dmitry , Levin David B , Wilkins John A , Sparling Richard
Published: September 2012

Background

Clostridium thermocellum produces H2 and ethanol, as well as CO2, acetate, formate, and lactate, directly from cellulosic biomass. It is therefore an attractive model for biofuel production via consolidated bioprocessing. Optimization of end-product yields and titres is crucial for making biofuel production economically feasible. Relative protein expression profiles may provide targets for metabolic engineering, while understanding changes in protein expression and metabolism in response to carbon limitation, pH, and growth phase may aid in reactor optimization. We performed shotgun 2D-HPLC-MS/MS on closed-batch cellobiose-grown exponential phase C. thermocellum cell-free extracts to determine relative protein expression profiles of core metabolic proteins involved carbohydrate utilization, energy conservation, and end-product synthesis. iTRAQ (isobaric tag for relative and absolute quantitation) based protein quantitation was used to determine changes in core metabolic proteins in response to growth phase.

Results

Relative abundance profiles revealed differential levels of putative enzymes capable of catalyzing parallel pathways. The majority of proteins involved in pyruvate catabolism and end-product synthesis were detected with high abundance, with the exception of aldehyde dehydrogenase, ferredoxin-dependent Ech-type [NiFe]-hydrogenase, and RNF-type NADH:ferredoxin oxidoreductase. Using 4-plex 2D-HPLC-MS/MS, 24% of the 144 core metabolism proteins detected demonstrated moderate changes in expression during transition from exponential to stationary phase. Notably, proteins involved in pyruvate synthesis decreased in stationary phase, whereas proteins involved in glycogen metabolism, pyruvate catabolism, and end-product synthesis increased in stationary phase. Several proteins that may directly dictate end-product synthesis patterns, including pyruvate:ferredoxin oxidoreductases, alcohol dehydrogenases, and a putative bifurcating hydrogenase, demonstrated differential expression during transition from exponential to stationary phase.

Conclusions

Relative expression profiles demonstrate which proteins are likely utilized in carbohydrate utilization and end-product synthesis and suggest that H2 synthesis occurs via bifurcating hydrogenases while ethanol synthesis is predominantly catalyzed by a bifunctional aldehyde/alcohol dehydrogenase. Differences in expression profiles of core metabolic proteins in response to growth phase may dictate carbon and electron flux towards energy storage compounds and end-products. Combined knowledge of relative protein expression levels and their changes in response to physiological conditions may aid in targeted metabolic engineering strategies and optimization of fermentation conditions for improvement of biofuels production.
Molecular epidemiology, antimicrobial susceptibilities and resistance mechanisms of Streptococcus pyogenes isolates resistant to erythromycin and tetracycline in Spain (1994–2006)
Authors: Rubio-López Virginia , Valdezate Sylvia , Álvarez David , Villalón Pilar , Medina María José , Salcedo Celia , Sáez-Nieto Juan-Antonio
Published: September 2012

Background

Group A Streptococcus (GAS) causes human diseases ranging in severity from uncomplicated pharyngitis to life-threatening necrotizing fasciitis and shows high rates of macrolide resistance in several countries. Our goal is to identify antimicrobial resistance in Spanish GAS isolates collected between 1994 and 2006 and to determine the molecular epidemiology (emm/T typing and PFGE) and resistance mechanisms of those resistant to erythromycin and tetracycline.

Results

Two hundred ninety-five out of 898 isolates (32.8%) were erythromycin resistant, with the predominance of emm 4T4, emm 75T25, and emm 28T28, accounting the 67.1% of the 21 emm/T types. Spread of emm 4T4, emm 75T25 and emm 28T28 resistant clones caused high rates of macrolide resistance. The distribution of the phenotypes was M (76.9%), cMLSB (20.3%), iMLSB (2.7%) with the involvement of the erythromycin resistance genes mef(A) (89.5%), msr(D) (81.7%), erm(B) (37.3%) and erm(A) (35.9%).

Sixty-one isolates were tetracycline resistant, with the main representation of the emm 77T28 among 20 emm/T types. To note, the combination of tet(M) and tet(O) tetracycline resistance genes were similar to tet(M) alone reaching values close to 40%. Resistance to both antibiotics was detected in 19 isolates of 7 emm/T types, being emm 11T11 and the cMLSB phenotype the most frequent ones. erm(B) and tet(M) were present in almost all the strains, while erm(A), mef(A), msr(D) and tet(O) appeared in less than half of them.

Conclusions

Spanish GAS were highly resistant to macrolides meanwhile showed minor resistance rate to tetracycline. A remarkable correlation between antimicrobial resistance and emm/T type was noticed. Clonal spread of emm 4T4, emm 75T25 and emm 28T28 was the main responsable for macrolide resistance where as that emm 77T28 clones were it to tetraclycline resistance. A wide variety of macrolide resistance genes were responsible for three macrolide resistance phenotypes.
Excessive proinflammatory cytokine and chemokine responses of human monocyte-derived macrophages to enterovirus 71 infection
Authors: Gong Xun , Zhou Jianfang , Zhu Wenfei , Liu Na , Li Jinsong , Li Lili , Jin Yu , Duan Zhaojun
Published: September 2012

Background

The levels of proinflammatory cytokine or chemokine in blood and cerebrospinal fluid are thought to be one of predictors for clinical severity of enterovirus 71 (EV71) infection, yet the cellular sources or signalling mechanism remain undefined. Here, we focused on the response of human primary monocyte-derived macrophages (MDMs) to EV71 virus and its possible mechanisms.

Methods

Human primary MDMs were infected by EV71 virus in vitro. Infectivity and viral replication were assayed, and cytokine responses were determined by Cytometric Bead Array(CBA) analysis. The relative changes of Toll-like receptors, retinoic acid-inducible gene I (RIG-I) and melamoma differentiation associated gene 5 (MDA5) mRNA expression were detected by real-time RT-PCR.

Results

Effective infection and viral replication were detected in EV71-infected MDMs. The titters of progeny virus released from EV71-infected MDMs gradually increased from 6-h to 48-h point of infection (POI.). Proinflammatory cytokines: IL-1, IL-6, TNF-α but not IFN-α and γ were induced in MDMs by EV71. EV71 infection significantly increased the release of IL-8, IP-10 and RANTES at 12-h or 24-h POI. Upregulation of TLR2, TLR7 and TLR8 mRNA expression rather than TLR3, TLR4, TLR6, TLR9, TLR10, RIG-I, MDA5 were found at different time points in EV71-infected MDMs.

Conclusions

Our findings suggested that macrophages are not only the important target cells but also the effectors during EV71 infection, and they may play an important role in the pathogenesis of EV71 infection. And the proinflammatory cytokine and chemokine responses in EV71-infected MDMs may be mediated by the activation of differential pattern of TLRs.
The changes of Th17 cells and the related cytokines in the progression of human colorectal cancers
Authors: Wang Jiansheng , Xu Kaiyu , Wu Jing , Luo Chenghua , Li Yuchen , Wu Xuebin , Gao Hong , Feng Guosheng , Yuan Bao-Zhu
Published: September 2012

Background

The role of Th17 cells in colorectal tumorigenesis and development still remains unclear, despite the fact that it has been established in the pathogenesis of autoimmune diseases.

Methods

We first analyzed Th17 cells and Treg cells using flow cytometry in the circulation of colorectal adenoma (CRA) and colorectal carcinoma (CRC) patients and healthy controls, and the frequency of Th17 cells in peripheral blood mononuclear cells (PBMCs) stimulated by anti-CD3 plus anti-CD28 and treated by IL-1β, IL-6, and TGF-β in different concentrations. We then detected cytokines IL-1β, IL-6, IL-17A, IL-21, IL-23 or TGF-β by ELISA in sera and supernatants from both normal and tumor tissues cultured ex vivo.

Results

It was found that the percentage of Th17 and Treg cells increased in the circulation of both CRA and CRC patients; the increase of Th17 cells in the circulation occurred in early stages, whereas the increase of Treg cells in the circulation and the increase of Th17 cells in tumor tissues occurred in advanced stages. The subsequent cytokine profiling showed that, along CRC progression, IL-1β, IL-17A and IL-23 underwent a similar change, while IL-6 in CRC exhibited an opposite change, with Th17 cells. In addition, high levels of TGF-β and IL-17A were detected in tumor tissues rather than in normal mucosa. The in vitro experiment further demonstrated that IL-1β, IL-6 or TGF-β modulated Th17 cell expansion in PBMC.

Conclusions

Our study reveals a unique change of Th17 cells, which is regulated possibly by IL-1β, IL-6 and TGF-β in the progression of CRC.
Short-chain fluorescent tryptophan tags for on-line detection of functional recombinant proteins
Authors: Siepert Eva-Maria , Gartz Esther , Tur Mehmet Kemal , Delbrück Heinrich , Barth Stefan , Büchs Jochen
Published: September 2012

Background

Conventional fluorescent proteins, such as GFP, its derivatives and flavin mononucleotide based fluorescent proteins (FbFPs) are often used as fusion tags for detecting recombinant proteins during cultivation. These reporter tags are state-of-the-art; however, they have some drawbacks, which can make on-line monitoring challenging. It is discussed in the literature that the large molecular size of proteins of the GFP family may stress the host cell metabolism during production. In addition, fluorophore formation of GFP derivatives is oxygen-dependent resulting in a lag-time between expression and fluorescence detection and the maturation of the protein is suppressed under oxygen-limited conditions. On the contrary, FbFPs are also applicable in an oxygen-limited or even anaerobic environment but are still quite large (58% of the size of GFP).

Results

As an alternative to common fluorescent tags we developed five novel tags based on clustered tryptophan residues, called W-tags. They are only 5-11% of the size of GFP. Based on the property of tryptophan to fluoresce in absence of oxygen it is reasonable to assume that the functionality of our W-tags is also given under anaerobic conditions. We fused these W-tags to a recombinant protein model, the anti-CD30 receptor single-chain fragment variable antibody (scFv) Ki-4(scFv) and the anti-MucI single-chain fragment variable M12(scFv). During cultivation in Microtiter plates, the overall tryptophan fluorescence intensity of all cultures was measured on-line for monitoring product formation via the different W-tags. After correlation of the scattered light signal representing biomass concentration and tryptophan fluorescence for the uninduced cultures, the fluorescence originating from the biomass was subtracted from the overall tryptophan signal. The resulting signal, thus, represents the product fluorescence of the tagged and untagged antibody fragments. The product fluorescence signal was increased. Antibodies with W-tags generated stronger signals than the untagged construct.

Conclusions

Our low-molecular-weight W-tags can be used to monitor the production of antibody fragments on-line. The binding specificity of the recombinant fusion protein is not affected, even though the binding activity decreases slightly with increasing number of tryptophan residues in the W-tags. Thus, the newly designed W-tags offer a versatile and generally applicable alternative to current fluorescent fusion tags.
Bipartite and tripartite Cucumber mosaic virus-based vectors for producing the Acidothermus cellulolyticus endo-1,4-β-glucanase and other proteins in non-transgenic plants
Authors: Hwang Min Sook , Lindenmuth Benjamin E , McDonald Karen A , Falk Bryce W
Published: September 2012

Background

Using plant viruses to produce desirable proteins in plants allows for using non-transgenic plant hosts and if necessary, the ability to make rapid changes in the virus construct for increased or modified protein product yields. The objective of this work was the development of advanced CMV-based protein production systems to produceAcidothermus cellulolyticusendo-1, 4-β-glucanase (E1) in non-transgenic plants.

Results

We used two newCucumber mosaic virus(CMV)-based vector systems for producing the green fluorescent protein (GFP) and more importantly, theAcidothermus cellulolyticusendo-1, 4-β-glucanase (E1) in non-transgenicNicotiana benthamianaplants. These are the inducibleCMVin(CMV-based inducible) and the autonomously replicatingCMVar(CMV-based advanced replicating) systems. We modified a binary plasmid containing the complete CMV RNA 3 cDNA to facilitate insertion of desired sequences, and to give modifications of the subgenomic mRNA 4 leader sequence yielding several variants. Quantitative RT-PCR and immunoblot analysis showed good levels of CMV RNA and coat protein accumulation for some variants of bothCMVinandCMVar. When genes for E1 or GFP were inserted in place of the CMV coat protein, both were produced in plants as shown by fluorescence (GFP) and immunoblot analysis. Enzymatic activity assays showed that active E1 was produced in plants with yields up to ~ 11 μg/g fresh weight (FW) for specific variant constructs. We also comparedin vitroCMV genomic RNA reassortants, and CMV RNA 3 mutants which lacked the C’ terminal 33 amino acids of the 3A movement protein in attempts to further increase E1 yield. Taken together specific variant constructs yielded up to ~21 μg/g FW of E1 in non-transgenic plants.

Conclusions

Intact, active E1 was rapidly produced in non-transgenic plants by using agroinfiltration with the CMV-based systems. This reduces the time and cost compared to that required to generate transgenic plants and still gives the comparable yields of active E1. Our modifications described here, including manipulating cloning sites for foreign gene introduction, enhance the ease of use. Also,N. benthamiana,which is particularly suitable for agroinfiltration, is a very good plant for transient protein production.
Higher normal ranges of urine albumin-to-creatinine ratio are independently associated with carotid intima-media thickness
Authors: Kweon Sun-Seog , Shin Min-Ho , Lee Young-Hoon , Choi Jin-Su , Nam Hae-Sung , Park Kyeong-Soo , Kim Do-Hyung , Jeong Seul-Ki
Published: September 2012

Background

High normal values of urine albumin-to-creatinine ratio (UACR) have been reported to have predictive values for hypertension, incident stroke, and higher mortality in the general population. This study aimed to investigate the association between normal ranges of UACR and carotid intima-media thickness (CIMT) in adult population.

Methods

We performed a cross-sectional study in adults aged 45 to 74 years who were living in Namwon City, South Korea. Both common CIMTs were measured, and mean values were calculated. Normal values of UACR were defined as <30mg/g and categorized into quintiles; less than 6.50, 6.51-9.79, 9.80-13.49, 13.50-18.89, and more than 18.90 mg/g. The association between the quintiles of UACR and common CIMT was analyzed and stratified by sex.

Results

A total of 7555 participants (3084 men and 4471 women) with normal UACR were enrolled in the present study. Common CIMT was positively and independently associated with increasing quintiles of UACR in men and women, even after adjusting for potential confounders including age and cardiovascular risk factors. Compared to the first quintile, the fifth quintile showed odds ratios of 1.80 (95% confidence intervals, 1.26-2.55) and 1.97 (1.28-3.04) for increased CIMT (>0.9mm) in men and women, respectively.

Conclusion

Higher UACR values within normal ranges (<30 mg/g) were positively and independently associated with CIMT in a Korean general population, suggesting that higher normal values of UACR might be a risk marker of subclinical carotid atherosclerosis.
Beat-to-beat estimation of the continuous left and right cardiac elastance from metrics commonly available in clinical settings
Authors: Stevenson David , Revie James , Chase J Geoffrey , Hann Christopher E , Shaw Geoffrey M , Lambermont Bernard , Ghuysen Alexandre , Kolh Philippe , Desaive Thomas
Published: September 2012

Introduction

Functional time-varying cardiac elastances (FTVE) contain a rich amount of information about the specific cardiac state of a patient. However, a FTVE waveform is very invasive to directly measure, and is thus currently not used in clinical practice. This paper presents a method for the estimation of a patient specific FTVE, using only metrics that are currently available in a clinical setting.

Method

Correlations are defined between invasively measured FTVE waveforms and the aortic and pulmonary artery pressures from 2 cohorts of porcine subjects, 1 induced with pulmonary embolism, the other with septic shock. These correlations are then used to estimate the FTVE waveform based on the individual aortic and pulmonary artery pressure waveforms, using the “other” dysfunction’s correlations as a cross validation.

Results

The cross validation resulted in 1.26% and 2.51% median errors for the left and right FTVE respectively on pulmonary embolism, while the septic shock cohort had 2.54% and 2.90% median errors.

Conclusions

The presented method accurately and reliably estimated a patient specific FTVE, with no added risk to the patient. The cross validation shows that the method is not dependent on dysfunction and thus has the potential for generalisation beyond pulmonary embolism and septic shock.
Effects of radiation emitted by WCDMA mobile phones on electromagnetic hypersensitive subjects
Authors: Kwon Min Kyung , Choi Joon Yul , Kim Sung Kean , Yoo Tae Keun , Kim Deok Won
Published: September 2012

Background

With the use of the third generation (3 G) mobile phones on the rise, social concerns have arisen concerning the possible health effects of radio frequency-electromagnetic fields (RF-EMFs) emitted by wideband code division multiple access (WCDMA) mobile phones in humans. The number of people with self-reported electromagnetic hypersensitivity (EHS), who complain of various subjective symptoms such as headache, dizziness and fatigue, has also increased. However, the origins of EHS remain unclear.

Methods

In this double-blind study, two volunteer groups of 17 EHS and 20 non-EHS subjects were simultaneously investigated for physiological changes (heart rate, heart rate variability, and respiration rate), eight subjective symptoms, and perception of RF-EMFs during real and sham exposure sessions. Experiments were conducted using a dummy phone containing a WCDMA module (average power, 24 dBm at 1950 MHz; specific absorption rate, 1.57 W/kg) within a headset placed on the head for 32 min.

Results

WCDMA RF-EMFs generated no physiological changes or subjective symptoms in either group. There was no evidence that EHS subjects perceived RF-EMFs better than non-EHS subjects.

Conclusions

Considering the analyzed physiological data, the subjective symptoms surveyed, and the percentages of those who believed they were being exposed, 32 min of RF radiation emitted by WCDMA mobile phones demonstrated no effects in either EHS or non-EHS subjects.
The pro-metastasis tyrosine phosphatase, PRL-3 (PTP4A3), is a novel mediator of oncogenic function of BCR-ABL in human chronic myeloid leukemia
Authors: Zhou Jianbiao , Cheong Lip-Lee , Liu Shaw-Cheng , Chong Phyllis SY , Mahara Sylvia , Bi Chonglei , Ong Kelly OK , Zeng Qi , Chng Wee Joo
Published: September 2012

Background

Resistance to tyrosine kinase inhibitors (TKIs) remains a challenge in management of patients with chronic myeloid leukemia (CML). A better understanding of the BCR-ABL signalling network may lead to better therapy.

Findings

Here we report the discovery of a novel downstream target of BCR-ABL signalling, PRL-3 (PTP4A3), an oncogenic tyrosine phosphatase. Analysis of CML cancer cell lines and CML patient samples reveals the upregulation of PRL-3. Inhibition of BCR-ABL signalling either by Imatinib or by RNAi silencing BCR-ABL reduces PRL-3 and increases cleavage of PARP. In contrast, the amount of PRL-3 protein remains constant or even increased in response to Imatinib treatment in drug resistant cells expressing P210 T315I. Finally, analysis with specific shRNA shows PRL-3 involvement in the proliferation and self-renewal of CML cells.

Conclusions

These data support a role for PRL-3 in BCR-ABL signalling and CML biology and may be a potential therapeutic target downstream of BCR-ABL in TKI resistant mutant cells.
Synthesis of 3D nanostructured metal alloy of immiscible materials induced by megahertz-repetition femtosecond laser pulses
Authors: Kiani Amirkianoosh , Waraich Palneet Singh , Venkatakrishnan Krishnan , Tan Bo
Published: September 2012

In this work, we have proposed a concept for the generation of three-dimensional (3D) nanostructured metal alloys of immiscible materials induced by megahertz-frequency ultrafast laser pulses. A mixture of two microparticle materials (aluminum and nickel oxide) and nickel oxide microparticles coated onto an aluminum foil have been used in this study. After laser irradiation, three different types of nanostructure composites have been observed: aluminum embedded in nickel nuclei, agglomerated chain of aluminum and nickel nanoparticles, and finally, aluminum nanoparticles grown on nickel microparticles. In comparison with current nanofabrication methods which are used only for one-dimensional nanofabrication, this technique enables us to fabricate 3D nanostructured metal alloys of two or more nanoparticle materials with varied composite concentrations under various predetermined conditions. This technique can lead to promising solutions for the fabrication of 3D nanostructured metal alloys in applications such as fuel-cell energy generation and development of custom-designed, functionally graded biomaterials and biocomposites.
Immunoglobulin G (IgG) attenuates neuroinflammation and improves neurobehavioral recovery after cervical spinal cord injury
Authors: Nguyen Dung Hoang , Cho Newton , Satkunendrarajah Kajana , Austin James W , Wang Jian , Fehlings Michael G
Published: September 2012

Background

Evidence suggests that the inflammatory events in the acute phase of spinal cord injury (SCI) exacerbate the initial trauma to the cord leading to poor functional recovery. As a result, minimizing the detrimental aspects of the inflammatory response after SCI is a promising treatment strategy. In this regard, immunoglobulin G (IgG) from pooled human serum is a promising treatment candidate. Due to its putative, though poorly characterized immuno-modulatory effects, IgG has been used clinically to treat neuroinflammatory disorders such as Guillain-Barré syndrome, but its effects in neurotrauma remain largely unexplored.

Methods

This study examines the potential neuroprotective effects of IgG in a well-characterized cervical model of SCI. Female Wistar rats were subject to moderate-severe clip compression injury at the C7-T1 level. IgG (0.4 g/kg) or saline was injected intravenously to randomly selected animals at 15 min post SCI. At several time points post SCI, biochemical assays, histology and immunohistochemistry analyses, and neurobehavioral assessments were used to examine the neuroprotective effects of IgG at the molecular, cellular, and neurobehavioral levels.

Results

We found that intravenous treatment of IgG following acute clip-compression SCI at C7-T1 significantly reduced two important inflammatory cytokines: interleukin (IL)-1β and IL-6. This early reduction in pro-inflammatory signaling was associated with significant reductions in neutrophils in the spinal cord and reductions in the expression of myeloperoxidase and matrix metalloproteinase-9 in the injured spinal cord at 24 h after SCI. These beneficial effects of IgG were associated with enhanced tissue preservation, improved neurobehavioral recovery as measured by the BBB and inclined plane tests, and enhanced electrophysiological evidence of central axonal conduction as determined by motor-evoked potentials.

Conclusion

The findings from this study indicate that IgG is a novel immuno-modulatory therapy which shows promise as a potential treatment for SCI.
Sublingual immunization with recombinant adenovirus encoding SARS-CoV spike protein induces systemic and mucosal immunity without redirection of the virus to the brain
Authors: Shim Byoung-Shik , Stadler Konrad , Nguyen Huan Huu , Yun Cheol-Heui , Kim Dong Wook , Chang Jun , Czerkinsky Cecil , Song Man Ki
Published: September 2012

Background

Sublingual (s.l.) administration of soluble protein antigens, inactivated viruses, or virus-like particles has been shown to induce broad immune responses in mucosal and extra-mucosal tissues. Recombinant replication-defective adenovirus vectors (rADVs) infect mucosa surface and therefore can serve as a mucosal antigen delivery vehicle. In this study we examined whether s.l. immunization with rADV encoding spike protein (S) (rADV-S) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) induces protective immunity against SARS-CoV and could serve as a safe mucosal route for delivery of rADV.

Results

Here, we show that s.l. administration of rADV-S induced serum SARS-CoV neutralizing and airway IgA antibodies in mice. These antibody responses are comparable to those induced by intranasal (i.n.) administration. In addition, s.l. immunization induced antigen-specific CD8+ T cell responses in the lungs that are superior to those induced by intramuscular immunization. Importantly, unlike i.n. administration, s.l. immunization with rADV did not redirect the rADV vector to the olfactory bulb.

Conclusion

Our study indicates that s.l. immunization with rADV-S is safe and effective in induction of a broad spectrum of immune responses and presumably protection against infection with SARS-CoV.