bluuebunny/crossref_metadata_2025 huggingface dataset:
Levels of von Willebrand factor and ADAMTS13 determine clinical outcome after cardioversion for atrial fibrillation
Authors: Bruno Veronika , Jarai Rudolf , Gruber Susanne , Höchtl Thomas , Brozovic Ivan , Farhan Serdar , Wojta Johann , Huber Kurt , Freynhofer Matthias
Published: December 2010

Von Willebrand factor (vWF) plays an essential role in platelet adhesion and thrombus formation. Patients with atrial fibrillation (AF) exhibit higher plasma vWF and lower ADAMTS13 antigen levels compared to controls. Little is known about vWF and ADAMTS13 in AF patients treated with cardioversion (CV). Thus we investigated the alterations of plasma vWF and ADAMTS13 after CV and evaluated the predictive value of these parameters for recurrence of AF. In this observational study we determined plasma levels of vWF and ADAMTS13 in 77 patients before and immediately after CV, as well as 24 hours (h) and six weeks thereafter, by means of commercially available assays. The vWF/ ADAMTS13-ratio was significantly elevated immediately after CV (p=0.02) and 24 h after CV (p=0.002) as compared to baseline levels. ADAMTS13, 24 h after CV, exhibited a significant association with recurrence of AF (HR: 0.97; p=0.037). Accordingly, tertiles of ADAMTS13 showed a stepwise inverse correlation with the risk of recurrent AF (HR: 0.50; p=0.009). After adjustment for confounders, ADAMTS13 remained significant as an independent predictor of recurrent AF (HR: 0.61; p=0.047). Similarly, the vWF/ADAMTS13-ratio, 24 h after CV, was associated with rhythm stability and remained an independent predictor of recurrent AF (HR: 1.88; p=0.028). The regulation of vWF and its cleaving protease ADAMTS13 after CV might play a critical role in producing a pro-thrombotic milieu immediately after CV for AF. Since ADAMTS13 plasma concentration and the vWF/ADAMTS13-ratio are independently associated with rhythm stability, these indexes might be used for prediction of recurrence of AF.
Total Anomalous Pulmonary Venous Connection
Authors: Seale Anna N. , Uemura Hideki , Webber Steven A. , Partridge John , Roughton Michael , Ho Siew Y. , McCarthy Karen P. , Jones Sheila , Shaughnessy Lynda , Sunnegardh Jan , Hanseus Katarina , Berggren Hakan , Johansson Sune , Rigby Michael L. , Keeton Barry R. , Daubeney Piers E.F. ,   on behalf of the British Congenital Cardiac Association
Published: December 2010

Background—

Late mortality after repair of total anomalous pulmonary venous connection is frequently associated with pulmonary venous obstruction (PVO). We aimed to describe the morphological spectrum of total anomalous pulmonary venous connection and identify risk factors for death and postoperative PVO.

Methods and Results—

We conducted a retrospective, international, collaborative, population-based study involving all 19 pediatric cardiac centers in the United Kingdom, Ireland, and Sweden. All infants with total anomalous pulmonary venous connection born between 1998 and 2004 were identified. Cases with functionally univentricular circulations or atrial isomerism were excluded. All available data and imaging were reviewed. Of 422 live-born cases, 205 (48.6%) had supracardiac, 110 (26.1%) had infracardiac, 67 (15.9%) had cardiac, and 37 (8.8%) had mixed connections. There were 2 cases (0.5%) of common pulmonary vein atresia. Some patients had extremely hypoplastic veins or, rarely, discrete stenosis of the individual veins. Sixty (14.2%) had associated cardiac anomalies. Sixteen died before intervention. Three-year survival for surgically treated patients was 85.2% (95% confidence interval 81.3% to 88.4%). Risk factors for death in multivariable analysis comprised earlier age at surgery, hypoplastic/stenotic pulmonary veins, associated complex cardiac lesions, postoperative pulmonary hypertension, and postoperative PVO. Sixty (14.8%) of the 406 patients undergoing total anomalous pulmonary venous connection repair had postoperative PVO that required reintervention. Three-year survival after initial surgery for patients with postoperative PVO was 58.7% (95% confidence interval 46.2% to 69.2%). Risk factors for postoperative PVO comprised preoperative hypoplastic/stenotic pulmonary veins and absence of a common confluence.

Conclusions—

Preoperative clinical and morphological features are important risk factors for postoperative PVO and survival.
Matricellular Signaling Molecule CCN1 Attenuates Experimental Autoimmune Myocarditis by Acting as a Novel Immune Cell Migration Modulator
Authors: Rother Madlen , Krohn Stefanie , Kania Gabriela , Vanhoutte Davy , Eisenreich Andreas , Wang Xiaomin , Westermann Dirk , Savvatis Kostas , Dannemann Nadine , Skurk Carsten , Hilfiker-Kleiner Denise , Cathomen Toni , Fechner Henry , Rauch Ursula , Schultheiss Heinz-Peter , Heymans Stephane , Eriksson Urs , Scheibenbogen Carmen , Poller Wolfgang
Published: December 2010

Background—

CCN1 is an evolutionary ancient matricellular protein that modulates biological processes associated with tissue repair. Induction at sites of injury was observed in conditions ranging from skin wounds to cardiac diseases, including ischemic and inflammatory cardiomyopathy. Here, we provide evidence of a novel function of CCN1 as a modulator of immune cell migration.

Methods and Results—

To understand the role of CCN1 in cardiomyopathies and to evaluate its therapeutic potential, we overexpressed CCN1 using an adenoviral hepatotropic vector in murine experimental autoimmune myocarditis, a model of human inflammatory cardiomyopathy. CCN1 gene transfer significantly reduced cardiac disease score and immune cell infiltration. In vivo tracking of hemagglutinin epitope–tagged CCN1 revealed binding to spleen macrophages but not to cardiomyocytes. Unexpectedly, CCN1 therapy left cardiac chemokine and cytokine expression unchanged but instead strongly inhibited the migration of spleen macrophages and lymphocytes, as evidenced by ex vivo transwell assays. In accordance with the ex vivo data, in vitro preincubation with CCN1 diminished transwell migration of human monocytes and abrogated their chemotactic response to monocyte chemoattractant protein-1, macrophage inflammatory protein-1α, and stromal cell–derived factor-1α. Further mechanistic studies showed that CCN1-driven modulation of immune cell migration is mimicked in part by cyclic RGD peptides currently in clinical evaluation for cancer therapy.

Conclusions—

Our proof-of-concept study suggests investigation of CCN1 as a novel, endogenous “parent compound” for chemotaxis modulation and of cyclic RGD peptides as a class of partially CCN1-mimetic drugs with immediate potential for clinical evaluation in cardiac diseases associated with chronic pathogenic inflammation.
Differences in Myocardial Structure and Coronary Microvasculature Between Men and Women With Coronary Artery Disease
Authors: Campbell Duncan J. , Somaratne Jithendra B. , Jenkins Alicia J. , Prior David L. , Yii Michael , Kenny James F. , Newcomb Andrew E. , Kelly Darren J. , Black Mary Jane
Published: December 2010

Women younger than 75 years with stable angina or acute coronary syndrome have higher cardiac mortality than similarly aged men, despite less obstructive coronary artery disease. To determine whether the myocardial structure and coronary microvasculature of women differs from that of men, we performed histological analysis of biopsies from nonischemic left ventricular myocardium from 46 men and 11 women undergoing coronary artery bypass graft surgery who did not have previous cardiac surgery, myocardial infarction, heart failure, atrial fibrillation, or furosemide therapy. The 2 patient groups had similar clinical characteristics, apart from a lower body surface area (BSA) in women (
P
=0.0015). Women had less interstitial fibrosis than men (
P
=0.019) but similar perivascular fibrosis. Arteriolar wall area/circumference ratio, a measure of arteriolar wall thickness, was 47% greater in women than men (
P
=0.012). Cardiomyocyte width and diffusion radius were positively correlated, and capillary length density was negatively correlated with BSA (
P
<0.05). Whereas cardiomyocyte width, capillary length density, diffusion radius, and cardiomyocyte width/BSA ratio were similar for men and women, women had a greater diffusion radius/BSA ratio (
P
=0.0038) and a greater diffusion radius/cardiomyocyte width ratio (
P
=0.027). Women also had lower vascular endothelial growth factor (VEGF) receptor-1 levels (
P
=0.048) and VEGF receptor-1/VEGF-A ratio (
P
=0.024) in plasma. We conclude that women with extensive coronary artery disease have greater arteriolar wall thickness and diffusion radius relative to BSA and to cardiomyocyte width than men, which may predispose to myocardial ischemia. Additional studies of larger numbers of women with less extensive coronary artery disease are required to confirm these findings.
Child labor in the People’s Republic of China: An ecological systems analysis
Authors: Minli Liao  , Jun Sung Hong
Published: December 2010

Child labor continues to be a serious social problem in China, and examination of the issue requires a contextualized understanding of the risk factors at various systems levels. We examine this issue using the perspective of Bronfenbrenner’s ecological theory. Implications for practice, policy and research are also discussed.
The multicultural paradox: The challenge of accommodating both power and trust in child protection
Authors: Johansson Ing-Marie
Published: December 2010

This research investigates whether ethnic background is under-communicated in child welfare services in the multicultural welfare state of Sweden, and if so how that affects the wellbeing of marginalized migrant families. It was found that a reciprocal and trustful relationship in combination with attention to the special needs of migrant families is a powerful vehicle for inclusion.
TEGDMA Reduces Mineralization in Dental Pulp Cells
Authors: Galler K.M. , Schweikl H. , Hiller K.-A. , Cavender A.C. , Bolay C. , D’Souza R.N. , Schmalz G.
Published: December 2010

Direct application of dentin bonding agents onto the exposed pulp has been advocated, but in vivo studies indicate a lack of reparative dentin formation. Our objective was to investigate the role of triethylene glycol dimethacrylate (TEGDMA), a commonly used compound in dentin bonding agents, as a potential inhibitor of mineralization. Human pulp cells were exposed to different concentrations of TEGDMA, and expression of the mineralization-related genes collagen I, alkaline phosphatase, bone sialoprotein, osteocalcin, Runx2, and dentin sialophosphoprotein was analyzed. Gene expression studies by real-time polymerase chain-reaction revealed a concentration- and time-dependent decrease of mineralization markers. A subtoxic TEGDMA concentration (0.3 mM) reduced expression levels by 5 to 20% after 4 hrs and by 50% after 12 hrs. Furthermore, alkaline phosphatase activity and calcium deposition were significantly lower in dental pulp cells treated with TEGDMA over 14 days. These findings indicate that even low TEGDMA concentrations might inhibit mineralization induced by dental pulp cells, thus impairing reparative dentin formation after pulp capping with dentin bonding agents.
The Relevance of Micro Places to Citywide Robbery Trends: A Longitudinal Analysis of Robbery Incidents at Street Corners and Block Faces in Boston
Authors: Braga Anthony A. , Hureau David M. , Papachristos Andrew V.
Published: December 2010

Robbery, and the fear it inspires, has a profound effect on the quality of life in certain urban neighborhoods. Recent advances in criminological research suggest that there is significant clustering of crime in micro places, or ‘‘hot spots,’’ that generate a disproportionate amount of criminal events in a city. In this article, the authors use growth curve regression models to uncover distinctive developmental trends in robbery incidents at street segments and intersections in Boston over a 29-year period. The authors find that robberies are highly concentrated at a small number of street segments and intersections rather than spread evenly across the urban landscape over the study time period. Roughly 1 percent and 8 percent of street segments and intersections in Boston are responsible for nearly 50 percent of all commercial robberies and 66 percent of all street robberies, respectively, between 1980 and 2008. Our findings suggest that citywide robbery trends may be best understood by examining micro-level trends at a relatively small number of places in urban environments.
The Northern Cities Shift in Chicago
Authors: McCarthy Corrine
Published: December 2010

This article examines the status of the Northern Cities Shift (NCS) as a change in progress in Chicago, the largest city within the Inland North. The progression of the NCS in Chicago and its social implementation have never been fully explored. Labov, Ash, and Boberg (2006) report that, for the Inland North as a whole, the NCS remains a vigorous change in progress. The present study finds only limited support for ongoing shifting within Chicago. Acoustic data from thirty-six speakers indicate that none of the three earliest shifting events, BAT, BOT, and BOUGHT, continues to shift in the expected direction in apparent time. Instead, BOT shows a marginal trend toward reversal of the NCS pattern. The three remaining vowels, BET, BIT, and BUT, shift in the expected direction. However, extensive phonetic overlap between BUT and BOUGHT is problematic for the NCS model as a single, integrated chain shift.
Article Commentary: Carbohydrate Intake and Overweight and Obesity Among Healthy Adults
Authors: Mullin Gerard E. , Cheskin Lawrence J.
Published: December 2010

Carbohydrate Intake and Overweight and Obesity Among Healthy Adults Merchant AT Vatanparast H Barlas S et al J Am Diet Assoc. 2009; 7:1165‐72.
Care of the insane in Lübeck during the 17th and 18th centuries
Authors: Dilling Horst , Thomsen Hans Peter , Hohagen Fritz
Published: December 2010

Only selected aspects of the history of the House of the Poor Insane in the Hanseatic Free City of Lübeck have been studied to date. This article presents the results of an entire source study of this small institution in the 17th and 18th centuries, and briefly also during the next 40 years after the opening of a new building. In addition to the minute-book of the Governors, now kept in the Lübeck Municipal Archives, the results are based primarily on the account-books, which illustrate the institution’s social history and activities. Examples are given. During most of the 17th century, the House was generally rather like a prison for the insane, but at the end of this century and in the early 18th there was a reform phase. This was followed by phases of repression and ‘containment’ at the end of the 18th century and in the early 19th century, before a renewed reform by the medical profession. The findings for Lübeck are compared with the development of inpatient care in institutions elsewhere, and the decisive factors in Lübeck are discussed.
The end of the asylum era in Central-Eastern Europe
Authors: Gazdag Gábor , Baran Brigitta , Rihmer Zoltán , Fink Max
Published: December 2010

As part of the structural change in the Hungarian mental health system, a decision was made in March 2007 to close the National Institute of Psychiatry and Neurology. Known by professionals and laymen as ‘Lipót’, it was the oldest psychiatric institute in Hungary. The purpose of this brief communication is to outline the history of the Institute, highlighting its role in the development and advancement of Hungarian psychiatry.
Cerebrospinal fluid CXCL13 in multiple sclerosis: a suggestive prognostic marker for the disease course
Authors: Khademi Mohsen , Kockum Ingrid , Andersson Magnus L , Iacobaeus Ellen , Brundin Lou , Sellebjerg Finn , Hillert Jan , Piehl Fredrik , Olsson Tomas
Published: December 2010

Background: Levels of CXCL13, a potent B-cell chemoattractant, are elevated in the cerebrospinal fluid (CSF) during multiple sclerosis (MS) and are associated with markers of MS activity. Levels decrease upon effective treatments.

Objective: Here we validate the potential role of CSF CXCL13 as a biomarker for aspects of MS in a large amount of clinical material, the majority collected at early diagnostic work-up.

Methods: CXCL13 was measured by ELISA in 837 subjects: relapsing–remitting MS (RRMS; n = 323), secondary progressive MS (SPMS; n = 40), primary progressive MS (PPMS; n = 24), clinically isolated syndrome (CIS; n = 79), other neurological diseases (ONDs; n = 181), ONDs with signs of inflammation or viral/bacterial infections (iONDs; n = 176) and healthy controls ( n = 14).

Results: Subjects with viral/bacterial infections had extremely high CXCL13 levels compared to all included groups ( p < 0.0001). CXCL13 was otherwise significantly higher in MS compared to the remaining controls ( p < 0.0001), and CIS ( p < 0.01). A significant and positive correlation between CXCL13 and relapse rate, the results obtained for the Expanded Disability Status Scale (EDSS) and the number of lesions detected by MRI was demonstrated. CXCL13 was increased in CIS conversion to clinically definite MS ( p < 0.001). Oligoclonal immunoglobulin band (OCB)-positive CIS or MS had significantly increased CXCL13 levels compared to OCB-negative CIS or MS ( p < 0.001 and p < 0.0001, respectively).

Conclusion: CXCL13 was associated with disease exacerbations and unfavourable prognosis in RRMS. Increased CXCL13 was not specific for MS since subjects with viral/bacterial infections exhibited even higher levels. High levels predicted CIS conversion to MS. We suggest that measurement of CSF CXCL13 can be part of the armamentarium in the diagnostic and prognostic work-up in MS and be of help in future treatment decisions.
Course of relapsing–remitting multiple sclerosis before, during and after natalizumab
Authors: Kaufman Michael D , Lee R , Norton HJ
Published: December 2010

The consequences of interruption of natalizumab treatment are incompletely known. The objective was to assess the confirmed annualized relapse rates for patients preceding initiation, during and following suspension of natalizumab therapy. A chart review was conducted and data were analyzed using the Generalized Estimating Equations. During natalizumab therapy the confirmed annualized relapse rate was 0.08, compared to 0.52 ( p = 0.0003) during the prior 12 months and to 0.35 ( p = 0.0032) during the following 3 to 24 months. Similar results were found when confirmed and unconfirmed were analyzed. To conclude, following cessation of natalizumab therapy disease activity rapidly returned to pre-natalizumab levels.
Disco edits and their discontents: The persistence of the analog in a digital era
Authors: Borschke Margie
Published: December 2010

This article foregrounds the distinction between two compositional forms and creative strategies in dance music — edits and remixes — as a way to gain a better understanding of the relationship between media use and media content, between producers and users, artifacts and events. It considers how the earliest disco edits in the 1970s were shaped by listeners (DJs and dancers) working in tandem with the material qualities and functional properties of vinyl records and other analog technologies and argues that while contemporary edits are made with digital tools, they continue to be in debt to their analog antecedents. In doing so this article critiques the enthusiastic adoption of ‘remix’ as a metaphor to describe digital culture and questions whether this rhetorical usage overshadows the aesthetic priorities and political implications of a variety of creative strategies that involve media use and re-use.
Orgies of the moment: Bataille’s anthropology of transgression and the defiance of danger in post-socialist Mozambique
Authors: Groes-Green Christian
Published: December 2010

In this article I explore socially marginalized young men’s excessive acts of violence, drug use, death race and unsafe sex against the background of George Bataille’s anthropology of transgression. When young men in the Mozambican capital engage in dangerous sex or violent riots, the findings indicate, it is less a sign of ignorance about HIV or indifference towards the rule of law than an expression of living in a ‘state of emergency’ where transgressive defiance of danger and death become attractive. Everyday transgressions of young men who call themselves moluwene (wild, unruly) are moulded in narratives and acts which at once oppose a smouldering socialist ideology of education and a neoliberal regime exiling marginalized young men from the realms of work and consumption to permanent unemployment, poverty and orgies of the moment.
Kindergarten theory: Childhood, affect, critical thought
Authors: Caselli Daniela
Published: December 2010

Current notions of affect are often underpinned by unacknowledged assumptions about spontaneity, materiality and immediacy. Childhood, which has traditionally been associated with these concepts (and for this reason has not been much debated within critical theory), helps us reconsider the political impact of affect theory. This is both because feminist theory has recently reconceptualized childhood and because positing affect as moments of intensity immanent to matter raises a number of problems from a feminist point of view. A passage from Mrs. Dalloway (illustrating how childhood works as a mimetic break within the project of literary modernism) will introduce an analysis of excerpts from Lisa Cartwright and Eve Kosofsky Sedgwick, two feminist case studies in which affect and childhood are linked. Referring to past psychoanalytical and political debates (especially by Jacqueline Rose, Juliet Mitchell and Walter Benjamin) that have anticipated some of the problems we are currently facing within feminist theory, the article investigates through childhood the politically problematic role of affect as ‘the new’ in critical theory.
Rationalizing seclusion: A preliminary analysis of a residential schooling scheme for poor girls in India
Authors: Balagopalan Sarada
Published: December 2010

The increased focus on issues of gender and schooling in India over the last decade has produced several gains that include more incentive schemes to make girls attend school, greater employment of women teachers and improved efforts to incorporate female protagonists in textbooks. However, a closer reading of this ‘gender’ focus reveals an inordinate concern with numbers, i.e. enrolment. The instrumentalism that underlies these efforts is revealed through a double-move effected by existing discourses. The first is to locate the reasons why girls are out of school strictly within a reading of cultural and familial practices and (secondly) to therefore fail to recognize normative practices of schooling and State policies as already deeply ‘gendered’. This double-move is epitomized in the Indian State’s more recent efforts to set up residential elementary schools for girls (Kasturba Gandhi Vidyalayas) in each district of the country; an effort that has been publicly lauded as the most effective way to overcome cultural barriers to girls’ schooling. Through a focus on policy documents that discuss this scheme, the article will interrogate the existing conflation of ‘gender’ with a biological/culturalist reading of the ‘girl child’. In what ways does this narrow focus naturalize a binary frame of reference around the traditional family/community and the empowered girl child? Why have State efforts around educating the ‘girl child’ not been subject to greater critical analysis amongst feminist scholars in India?
Mapping Mental Function to Brain Structure: How Can Cognitive Neuroimaging Succeed?
Authors: Poldrack Russell A.
Published: December 2010

The goal of cognitive neuroscience is to identify the mapping between brain function and mental processing. In this article, I examine the strategies that have been used to identify such mappings and argue that they may be fundamentally unable to identify selective structure–function mappings. To understand the functional anatomy of mental processes, it will be necessary for researchers to move from the brain-mapping strategies that the field has employed toward a search for selective associations. This will require a greater focus on the structure of cognitive processes, which can be achieved through the development of formal ontologies that describe the structure of mental processes. In this article, I outline the Cognitive Atlas Project, which is developing such ontologies, and show how this knowledge could be used in conjunction with data-mining approaches to more directly relate mental processes and brain function.
Frontiers in Human Neuroscience: The Golden Triangle and Beyond
Authors: Decety Jean , Cacioppo John
Published: December 2010

The development and application of neuroimaging methods offers powerful means to study brain functions, but the resulting knowledge is more likely to be beneficial when combined with conceptual analyses that decompose complex psychological constructs into component structures, representations, processes, and computations; converging measures that gauge neural events at different temporal and spatial scales; behavioral measures that permit fine-grain analyses of brain–behavior associations; and experimental (e.g., lesion studies and/or transcranial magnetic stimulation) and nonhuman animal studies that test the putative role of specific brain structures, circuits, or processes. In addition, quantitative meta-analyses are important to move beyond idiosyncrasies of individual studies, and neurodevelopmental investigations can contribute to our understanding of brain–behavior associations.
Pathophysiology of sickle cell disease is mirrored by the red blood cell metabolome
Authors: Darghouth Dhouha , Koehl Bérengère , Madalinski Geoffrey , Heilier Jean-François , Bovee Petra , Xu Ying , Olivier Marie-Françoise , Bartolucci Pablo , Benkerrou Malika , Pissard Serge , Colin Yves , Galacteros Frederic , Bosman Giel , Junot Christophe , Roméo Paul-Henri
Published: December 2010

Emerging metabolomic tools can now be used to establish metabolic signatures of specialized circulating hematopoietic cells in physiologic or pathologic conditions and in human hematologic diseases. To determine metabolomes of normal and sickle cell erythrocytes, we used an extraction method of erythrocytes metabolites coupled with a liquid chromatography-mass spectrometry–based metabolite profiling method. Comparison of these 2 metabolomes identified major changes in metabolites produced by (1) endogenous glycolysis characterized by accumulation of many glycolytic intermediates; (2) endogenous glutathione and ascorbate metabolisms characterized by accumulation of ascorbate metabolism intermediates, such as diketogulonic acid and decreased levels of both glutathione and glutathione disulfide; (3) membrane turnover, such as carnitine, or membrane transport characteristics, such as amino acids; and (4) exogenous arginine and NO metabolisms, such as spermine, spermidine, or citrulline. Finally, metabolomic analysis of young and old normal red blood cells indicates metabolites whose levels are directly related to sickle cell disease. These results show the relevance of metabolic profiling for the follow-up of sickle cell patients or other red blood cell diseases and pinpoint the importance of metabolomics to further depict the pathophysiology of human hematologic diseases.
The effects of viral load on pseudorabies virus gene expression
Authors: Tóth Judit S , Tombácz Dóra , Takács Irma F , Boldogkői Zsolt
Published: December 2010

Background

Herpesvirus genes are classified into distinct kinetic groups on the basis of their expression dynamics during lytic growth of the virus in cultured cells at a high, typically 10 plaque-forming units/cell multiplicity of infection (MOI). It has been shown that both the host response and the success of a pathogen are dependent on the quantity of particles infecting an organism. This work is a continuation of an earlier study [1], in which we characterized the overall expression of PRV genes following low-MOI infection. In the present study, we have addressed the question of whether viral gene expressions are dependent on the multiplicity of infection by comparing gene expressions under low and high-MOI conditions.

Results

In the present study, using a real-time RT-PCR assay, we address the question of whether the expression properties of the pseudorabies virus (PRV) genes are dependent on the number of virion particles infecting a single cell in a culture. Our analysis revealed a significant dependence of the gene expression on the MOI in most of these genes. Specifically, we found that most of the examined viral genes were expressed at a lower level at a low MOI (0.1) than at a high MOI (10) experiment in the early stage of infection; however, this trend reversed by six hour post-infection in more than half of the genes. Furthermore, in the high-MOI infection, several PRV genes substantially declined within the 4 to 6-h infection period, which was not the case in the low-MOI infection. In the low-MOI infection, the level of antisense transcript (AST), transcribed from the antiparallel DNA strand of the immediate-early 180 (ie180) gene, was comparable to that of ie180 mRNA, while in the high-MOI experiment (despite the 10 times higher copy number of the viral genome in the infected cells) the amount of AST dropped by more than two log values at the early phase of infection. Furthermore, our analysis suggests that adjacent PRV genes are under a common regulation. This is the first report on the effect of the multiplicity of infection on genome-wide gene expression of large DNA viruses, including herpesviruses.

Conclusion

Our results show a strong dependence of the global expression of PRV genes on the MOI. Furthermore, our data indicate a strong interrelation between the expressions of ie180 mRNA and AST, which determines the expression properties of the herpesvirus genome and possibly the replication strategy (lytic or latent infection) of the virus in certain cell types.
Complement in patients receiving maintenance hemodialysis: functional screening and quantitative analysis
Authors: Inoshita Hiroyuki , Ohsawa Isao , Kusaba Gaku , Ishii Masaya , Onda Kisara , Horikoshi Satoshi , Ohi Hiroyuki , Tomino Yasuhiko
Published: December 2010

Background

The complement system is vital for innate immunity and is implicated in the pathogenesis of inflammatory diseases and the mechanism of host defense. Complement deficiencies occasionally cause life-threatening diseases. In hemodialysis (HD) patients, profiles on complement functional activity and deficiency are still obscure. The objectives of the present study were to measure the functional complement activities of the classical pathway (CP), lectin pathway (LP) and alternative pathway (AP) using a novel method and consequently to elucidate the rates of deficiencies among HD patients.

Methods

In the present study, 244 HD patients at one dialysis center and 204 healthy controls were enrolled. Functional complement activities were measured simultaneously using the Wielisa®-kit. The combination of the results of these three pathway activities allows us to speculate which candidate complement is deficient; subsequently, the deficient complement was determined.

Results

All three functional complement activities were significantly higher in the HD patients than in the control group (P < 0.01 for all cases). After identifying candidates in both groups with complement deficiencies using the Wielisa®-kit, 16 sera (8.8%) with mannose-binding lectin (MBL) deficiency, 1 serum (0.4%) with C4 deficiency, 1 serum (0.4%) with C9 deficiency, and 1 serum (0.4%) with B deficiency were observed in the HD group, and 18 sera (8.8%) with MBL deficiency and 1 serum (0.5%) with B deficiency were observed in the control group. There were no significant differences in the 5-year mortality rate between each complement-deficient group and the complement-sufficient group among the HD patients.

Conclusion

This is the first report that profiles complement deficiencies by simultaneous measurement of functional activities of the three complement pathways in HD patients. Hemodialysis patients frequently suffer from infections or malignancies, but functional complement deficiencies do not confer additional risk of mortality.
Design of the EXercise Intervention after Stem cell Transplantation (EXIST) study: a randomized controlled trial to evaluate the effectiveness and cost-effectiveness of an individualized high intensity physical exercise program on fitness and fatigue in patients with multiple myeloma or (non-) Hodgkin's lymphoma treated with high dose chemotherapy and autologous stem cell transplantation
Authors: Persoon Saskia , Kersten Marie José , ChinAPaw Mai JM , Buffart Laurien M , Burghout Heleen , Schep Goof , Brug Johannes , Nollet Frans
Published: December 2010

Background

The use of high-dose chemotherapy combined with autologous stem cell transplantation has improved the outcome of hematologic malignancies. Nevertheless, this treatment can cause persistent fatigue and a reduced global quality of life, role and physical function. Physical exercise interventions may be beneficial for physical fitness, fatigue and quality of life. However, the trials conducted so far to test the effects of physical exercise interventions in this group of patients were of poor to moderate methodological quality and economic evaluations are lacking. Hence there is need for a rigorous, appropriately controlled assessment of the effectiveness of exercise programs in these patients. The aims of the present study are (1) to determine the effectiveness of an individualized high intensity strength and interval training program with respect to physiological and psychological health status in patients with multiple myeloma or (non-)Hodgkin's lymphoma who have recently undergone high dose chemotherapy followed by autologous stem cell transplantation; and (2) to evaluate the cost-effectiveness of this program.

Methods

A multicenter, prospective, single blind randomized controlled trial will be performed. We aim to recruit 120 patients within an inclusion period of 2 years at 7 hospitals in the Netherlands. The patients will be randomly assigned to one of two groups: (1) intervention plus usual care; or (2) usual care. The intervention consists of an 18-week individualized supervised high-intensity exercise program and counselling. The primary outcomes (cardiorespiratory fitness, muscle strength and fatigue) and secondary outcomes are assessed at baseline, at completion of the intervention and at 12 months follow-up.

Discussion

The strengths of this study include the solid trial design with clearly defined research groups and standardized outcome measures, the inclusion of an economic evaluation and the inclusion of both resistance and endurance exercise in the intervention program.

Trial registration

This study is registered at the Netherlands Trial Register (NTR2341)
DNAzyme-mediated recovery of small recombinant RNAs from a 5S rRNA-derived chimera expressed in Escherichia coli
Authors: Liu Yamei , Stepanov Victor G , Strych Ulrich , Willson Richard C , Jackson George W , Fox George E
Published: December 2010

Background

Manufacturing large quantities of recombinant RNAs by overexpression in a bacterial host is hampered by their instability in intracellular environment. To overcome this problem, an RNA of interest can be fused into a stable bacterial RNA for the resulting chimeric construct to accumulate in the cytoplasm to a sufficiently high level. Being supplemented with cost-effective procedures for isolation of the chimera from cells and recovery of the recombinant RNA from stabilizing scaffold, this strategy might become a viable alternative to the existing methods of chemical or enzymatic RNA synthesis.

Results

Sequence encoding a 71-nucleotide recombinant RNA was inserted into a plasmid-borne deletion mutant of theVibrio proteolyticus5S rRNA gene in place of helix III - loop C segment of the original 5S rRNA. After transformation intoEscherichia coli, the chimeric RNA (3×penaRNA) was expressed constitutively fromE. coli rrnBP1 and P2 promoters. The RNA chimera accumulated to levels that exceeded those of the host's 5S rRNA. A novel method relying on liquid-solid partitioning of cellular constituents was developed for isolation of total RNA from bacterial cells. This protocol avoids toxic chemicals, and is therefore more suitable for large scale RNA purification than traditional methods. A pair of biotinylated 8-17 DNAzymes was used to bring about the quantitative excision of the 71-nt recombinant RNA from the chimera. The recombinant RNA was isolated by sequence-specific capture on beads with immobilized complementary deoxyoligonucleotide, while DNAzymes were recovered by biotin affinity chromatography for reuse.

Conclusions

The feasibility of a fermentation-based approach for manufacturing large quantities of small RNAsin vivousing a "5S rRNA scaffold" strategy is demonstrated. The approach provides a route towards an economical method for the large-scale production of small RNAs including shRNAs, siRNAs and aptamers for use in clinical and biomedical research.
Thymoglobulin, interferon-γ and interleukin-2 efficiently expand cytokine-induced killer (CIK) cells in clinical-grade cultures
Authors: Bonanno Giuseppina , Iudicone Paola , Mariotti Andrea , Procoli Annabella , Pandolfi Annino , Fioravanti Daniela , Corallo Maria , Perillo Alessandro , Scambia Giovanni , Pierelli Luca , Rutella Sergio
Published: December 2010

Background

Cytokine-induced killer (CIK) cells are typically differentiatedin vitrowith interferon (IFN)-γ and αCD3 monoclonal antibodies (mAb), followed by the repeated provision of interleukin (IL)-2. It is presently unknown whether thymoglobulin (TG), a preparation of polyclonal rabbit γ immunoglobulins directed against human thymocytes, can improve the generation efficiency of CIK cells compared with αCD3 mAb in a clinical-grade culture protocol.

Methods

Peripheral blood mononuclear cells (PBMC) from 10 healthy donors and 4 patients with solid cancer were primed with IFN-γ on day 0 and low (50 ng/ml), intermediate (250 ng/ml) and high (500 ng/ml) concentrations of either αCD3 mAb or TG on day 1, and were fed with IL-2 every 3 days for 21 days. Aliquots of cells were harvested weekly to monitor the expression of representative members of the killer-like immunoglobulin receptor (KIR), NK inhibitory receptor, NK activating receptor and NK triggering receptor families. We also quantified the frequency ofbona fideregulatory T cells (Treg), a T-cell subset implicated in the down-regulation of anti-tumor immunity, and tested thein vitrocytotoxic activity of CIK cells against NK-sensitive, chronic myeloid leukaemia K562 cells.

Results

CIK cells expanded more vigorously in cultures supplemented with intermediate and high concentrations of TG compared with 50 ng/ml αCD3 mAb. TG-driven CIK cells expressed a constellation of NK activating/inhibitory receptors, such as CD158a and CD158b, NKp46, NKG2D and NKG2A/CD94, released high quantities of IL-12p40 and efficiently lysed K562 target cells. Of interest, the frequency of Treg cells was lower at any time-point compared with PBMC cultures nurtured with αCD3 mAb. Cancer patient-derived CIK cells were also expanded after priming with TG, but they expressed lower levels of the NKp46 triggering receptor and NKG2D activating receptor, thus manifesting a reduced ability to lyse K562 cells.

Conclusions

TG fosters the generation of functional CIK cells with no concomitant expansion of tumor-suppressive Treg cells. The culture conditions described herein should be applicable to cancer-bearing individuals, although the differentiation of fully functional CIK cells may be hindered in patients with advanced malignancies.
Transforming growth factor-β1 induces matrix metalloproteinase-9 and cell migration in astrocytes: roles of ROS-dependent ERK- and JNK-NF-κB pathways
Authors: Hsieh Hsi-Lung , Wang Hui-Hsin , Wu Wen-Bin , Chu Po-Ju , Yang Chuen-Mao
Published: December 2010

Background

Transforming growth factor-β (TGF-β) and matrix metalloproteinases (MMPs) are the multifunctional factors during diverse physiological and pathological processes including development, wound healing, proliferation, and cancer metastasis. Both TGF-β and MMPs have been shown to play crucial roles in brain pathological changes. Thus, we investigated the molecular mechanisms underlying TGF-β1-induced MMP-9 expression in brain astrocytes.

Methods

Rat brain astrocytes (RBA-1) were used. MMP-9 expression was analyzed by gelatin zymography and RT-PCR. The involvement of signaling molecules including MAPKs and NF-κB in the responses was investigated using pharmacological inhibitors and dominant negative mutants, determined by western blot and gene promoter assay. The functional activity of MMP-9 was evaluated by cell migration assay.

Results

Here we report that TGF-β1 induces MMP-9 expression and enzymatic activity via a TGF-β receptor-activated reactive oxygen species (ROS)-dependent signaling pathway. ROS production leads to activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and c-Jun-N-terminal kinase (JNK) and then activation of the NF-κB transcription factor. Activated NF-κB turns on transcription of the MMP-9 gene. The rat MMP-9 promoter, containing a NF-κB cis-binding site, was identified as a crucial domain linking to TGF-β1 action.

Conclusions

Collectively, in RBA-1 cells, activation of ERK1/2- and JNK-NF-κB cascades by a ROS-dependent manner is essential for MMP-9 up-regulation/activation and cell migration induced by TGF-β1. These findings indicate a new regulatory pathway of TGF-β1 in regulating expression of MMP-9 in brain astrocytes, which is involved in physiological and pathological tissue remodeling of central nervous system.
A single-amino-acid substitution in the HA protein changes the replication and pathogenicity of the 2009 pandemic A (H1N1) influenza viruses in vitro and in vivo
Authors: Xu Lili , Bao Linlin , Lv Qi , Deng Wei , Ma Yila , Li Fengdi , Zhan Lingjun , Zhu Hua , Ma Chunmei , Qin Chuan
Published: December 2010

Background

The novel pandemic A (H1N1) virus was first identified in Mexico in April 2009 and since then it spread world wide over a short period of time. Although the virus infection is generally associated with mild disease and a relatively low mortality, it is projected that mutations in specific regions of the viral genome, especially within the receptor binding domain of the hemagglutinin (HA) protein could result in more virulent virus stains, leading to a more severe pandemic.

Results

Here, we found that a single amino acid substitution of Asp-to-Gly at position 222 in the HA protein of the A (H1N1) virus occurred after two passage propagation in the allantoic cavities of chicken embryonated eggs, and this single residue variation dramatically increased the viral replication ability in MDCK cells and pathogenicity in BALB/c mice.

Conclusions

A substitution of Asp-to-Gly at position 222 in the HA protein was prone to occur under positive selection pressures, and this single amino acid mutation could dramatically increase the virus replication ability in vitro and pathogenicity in vivo. Our finding offers a better understanding of the transmission and evolution of the 2009 pandemic A (H1N1) virus and brings attention to further potentially severe influenza pandemic that may result from cross-host evolution of the influenza viruses.
Differential expression of microRNAs in GH-secreting pituitary adenomas
Authors: Mao Zhi-Gang , He Dong-Sheng , Zhou Jing , Yao Bin , Xiao Wei-Wei , Chen Chun-Hua , Zhu Yong-Hong , Wang Hai-Jun
Published: December 2010

Background

The purpose of this study was to (1) identify specific miRNAs in growth hormones (GH)-secreting pituitary adenomas; (2) determine the relationship between the expression of these miRNAs and tumor size, somatostatin analogs treatment, and responsiveness to somatostatin analogs (SSA).

Methods

Fifteen GH-secreting adenomas patients were treated with lanreotide for 4 months before surgery. Patients with 50% reduction of GH secretion by lanreotide were considered as SSA responders, while patients with less than 50% of GH reduction were considered as SSA nonresponders. We analyzed the miRNAs in 21 GH-secreting pituitary adenomas and 6 normal pituitaries by miRCURY™ LNA array and some differentially expressed miRNAs were validated by quantitative real-time PCR.

Results

Fifty-two miRNAs were differentially expressed between GH-secreting pituitary adenomas and normal pituitaries. Differential expression of 9 miRNAs was observed between micro- and macro-adenomas. Thirteen miRNAs were differentially expressed between tumor samples from lanreotide-treated patients and those from lanreotide-untreated patients. Seven miRNAs were differentially expressed between SSA responders or GH nonresponders. Several identified miRNAs may be involved in cell proliferation, apoptosis, cancer development and progression.

Conclusions

Our results indicate that altered miRNAs expression is involved in GH-secreting pituitary adenomas transformation, which will shed light on the mechanisms for the treatment of acromegaly by SSA. Identification and characterization of the targets of altered miRNAs genes may elucidate molecular mechanisms involved in the pathogenesis of pituitary adenoma.
HCMV pUS28 initiates pro-migratory signaling via activation of Pyk2 kinase
Authors: Vomaske Jennifer , Varnum Susan , Melnychuk Ryan , Smith Patricia , Pasa-Tolic Ljiljana , Shutthanandan Janani I , Streblow Daniel N
Published: December 2010

Background

Human Cytomegalovirus (HCMV) has been implicated in the acceleration of vascular disease and chronic allograft rejection. Recently, the virus has been associated with glioblastoma and other tumors. We have previously shown that the HCMV-encoded chemokine receptor pUS28 mediates smooth muscle cell (SMC) and macrophage motility and this activity has been implicated in the acceleration of vascular disease. pUS28 induced SMC migration involves the activation of the protein tyrosine kinases (PTKs) Src and Focal adhesion kinase as well as the small GTPase RhoA. The PTK Pyk2 has been shown to play a role in cellular migration and formation of cancer, especially glioblastoma. The role of Pyk2 in pUS28 signaling and migration are unknown.

Methods

In the current study, we examined the involvement of the PTK Pyk2 in pUS28-induced cellular motility. We utilized in vitro migration of SMC to determine the requirements for Pyk2 in pUS28 pro-migratory signaling. We performed biochemical analysis of Pyk2 signaling in response to pUS28 activation to determine the mechanisms involved in pUS28 migration. We performed mass spectrometric analysis of Pyk2 complexes to identify novel Pyk2 binding partners.

Results

Expression of a mutant form of Pyk2 lacking the autophosphorylation site (Tyr-402) blocks pUS28-mediated SMC migration in response to CCL5, while the kinase-inactive Pyk2 mutant failed to elicit the same negative effect on migration. pUS28 stimulation with CCL5 results in ligand-dependent and calcium-dependent phosphorylation of Pyk2 Tyr-402 and induced the formation of an active Pyk2 kinase complex containing several novel Pyk2 binding proteins. Expression of the autophosphorylation null mutant Pyk2 F402Y did not abrogate the formation of an active Pyk2 kinase complex, but instead prevented pUS28-mediated activation of RhoA. Additionally, pUS28 activated RhoA via Pyk2 in the U373 glioblastoma cells. Interestingly, the Pyk2 kinase complex in U373 contained several proteins known to participate in glioma tumorigenesis.

Conclusions

These findings represent the first demonstration that pUS28 signals through Pyk2 and that this PTK participates in pUS28-mediated cellular motility via activation of RhoA. Furthermore, these results provide a potential mechanistic link between HCMV-pUS28 and glioblastoma cell activation.