bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Supplemental Intra-articular Lubricin and Hyaluronic Acid on the Progression of Posttraumatic Arthritis in the Anterior Cruciate Ligament–Deficient Rat Knee
Authors: Teeple Erin , Elsaid Khaled A. , Jay Gregory D. , Zhang Ling , Badger Gary J. , Akelman Matthew , Bliss Thomas F. , Fleming Braden C.
Published: September 2010

Background: Lubricin and hyaluronic acid lubricate articular cartilage and prevent wear. Because lubricin loss occurs after anterior cruciate ligament injury, intra-articular lubricin injections may reduce cartilage damage in the anterior cruciate ligament–deficient knee.

Purpose: This study was conducted to determine if lubricin and/or hyaluronic acid supplementation will reduce cartilage damage in the anterior cruciate ligament–deficient knee.

Study Design: Controlled laboratory study.

Methods: Thirty-six male rats, 3 months old, underwent unilateral anterior cruciate ligament transection. They were randomized to 4 treatments: (1) saline (phosphate-buffered saline [PBS]), (2) hyaluronic acid (HA), (3) purified human lubricin (LUB), and (4) LUB and HA (LUB+HA). Intra-articular injections were given twice weekly for 4 weeks starting 1 week after surgery. Knees were harvested 1 week after the final injection. Radiographs of each limb and synovial fluid lavages were obtained at harvest. Histologic analysis was performed to assess cartilage damage using safranin O/fast green staining. Radiographs were scored for the severity of joint degeneration using the modified Kellgren-Lawrence scale. Synovial fluid levels of sulfated glycosaminoglycan, collagen II breakdown, interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α), and lubricin were measured using enzyme-linked immunosorbent assay (ELISA).

Results: Treatment with LUB or LUB+HA significantly decreased radiographic and histologic scores of cartilage damage ( P = .039 and P = .015, respectively) when compared with the PBS and HA conditions. There was no evidence of an effect of HA nor was the LUB effect HA-dependent, suggesting that the addition of HA did not further reduce damage. The synovial fluid of knees treated with LUB had significantly more lubricin in the synovial fluid at euthanasia, although there were no differences in the other cartilage metabolism biomarkers.

Conclusion: Supplemental intra-articular LUB reduced cartilage damage in the anterior cruciate ligament–transected rat knee 6 weeks after injury, while treatment with HA did not.

Clinical Relevance: Although longer term studies are needed, intra-articular supplementation (tribosupplementation) with lubricin after anterior cruciate ligament injury may protect the articular cartilage in the anterior cruciate ligament–injured knee.
Shockwave Therapy for the Treatment of Chronic Proximal Hamstring Tendinopathy in Professional Athletes
Authors: Cacchio Angelo , Rompe Jan D. , Furia John P. , Susi Piero , Santilli Valter , De Paulis Fosco
Published: September 2010

Background: Chronic proximal hamstring tendinopathy is an overuse syndrome that is usually managed by nonoperative methods. Shockwave therapy has proved to be effective in many tendinopathies.

Hypothesis: Shockwave therapy may be more effective than other nonoperative treatments for chronic proximal hamstring tendinopathy.

Study Design: Randomized controlled clinical study; Level of evidence, 1.

Methods: Forty professional athletes with chronic proximal hamstring tendinopathy were enrolled between February 1, 2004, and September 30, 2006. Patients were randomly assigned to receive either shockwave therapy, consisting of 2500 impulses per session at a 0.18 mJ/mm2 energy flux density without anesthesia, for 4 weeks (SWT group, n = 20), or traditional conservative treatment consisting of nonsteroidal anti-inflammatory drugs, physiotherapy, and an exercise program for hamstring muscles (TCT group, n = 20). Patients were evaluated before treatment, and 1 week and 3, 6, and 12 months after the end of treatment. The visual analog scale (VAS) score for pain and Nirschl phase rating scale (NPRS) were used as primary outcome measures.

Results: The patients were observed for a mean of 10.7 months (range, 1-12 months). Six patients were lost to follow-up because they underwent a surgical intervention: 3 (all in TCT group) were lost at 3 months; 2 (1 in each group), at 6 months; and 1 (in the TCT group), at 12 months. Primary follow-up was at 3 months after the beginning of treatment. The VAS scores in the SWT and TCT groups were 7 points before treatment ( P = .84), and 2 points and 5 points, respectively, 3 months after treatment ( P < .001). The NPRS scores in the SWT and TCT groups were 5 points in either group before treatment ( P = .48), and 2 points and 6 points, respectively, 3 months after treatment ( P < .001). At 3 months after treatment, 17 of the 20 patients (85%) in the SWT group and 2 of the 20 patients (10%) in the TCT group achieved a reduction of at least 50% in pain ( P < .001). There were no serious complications in the SWT group.

Conclusion: Shockwave therapy is a safe and effective treatment for patients with chronic proximal hamstring tendinopathy.
A methodology for developing strategic municipal solid waste management plans with an application in Greece
Authors: Economopoulos AP
Published: September 2010

A rational approach for developing optimal municipal solid waste (MSW) management plans comprises the strategic and the detailed planning phases. The present paper focuses on the former, the objective of which is to screen management alternatives so as to select the ones that are able to fulfil all legal and other management requirements with reasonable cost. The analysis considers the transportation, treatment and final disposal of the commingled wastes that remain after the application of material recovery at the source programmes and comprises 10 elements, four of which are region-dependent and the remaining ones application-dependent. These elements and their inter-dependencies are described and the entire methodology is applied to Greece. The application considers the existing regional plans and shows that they are incompatible with the existing EU Directives, as well as overly expensive. To address this problem, a new plan is developed in accordance with the rational planning principles of the present methodology. The comparative evaluation of the above alternatives shows that the existing regional plans, in addition to being incompatible with the applicable EU Directives, require 4.3 to 4.8 times (3.7 to 4.4 billion ) higher capital investment and their annual cost is at least 2.1 to 2.3 times (590 to 735 million year —1) higher in comparison with the new national plan.
Sustainable landfilling in tropical conditions: comparison between open and closed cell approach
Authors: Visvanathan C. , Karthikeyan Obuli P , Park KH
Published: September 2010

Two landfill test cells were constructed in a tropical climate using locally available low-cost materials. One cell was operated without a cover on the municipal solid waste to simulate ‘open landfill’ conditions and the other cell was covered to create ‘closed landfill’ conditions. Both test cells were monitored over a period of 290 days under rainy, dry and artificial wetting conditions. Due to the relatively high compaction density of waste in the closed test cell, the substrate settlement was gradual and comparatively lower than in the open test cell. Multiple top covers in the closed test cell resulted in significant run-off of incident precipitation during the rainy season, which delayed the waste stabilization and subsequently produced a lesser volume of leachate. On the other hand, operation of the open test cell was found to be advantageous in terms of leachate management and substrate settlement along with waste stabilization pattern. Infiltration of rain-water into the waste mass leached out the maximum organic pollutants and oxidized the nitrogen content, which is deemed to be a benefit of operating an open cell landfill under tropical conditions. Artificial wetting during dry periods by recirculation of stored leachate notably accelerated the waste stabilization and secondary substrate settlement in the open test cell. The continuous monitoring of ground-water quality from the site showed only seasonal variations.
Characterization of immune cell subsets during the active phase of multiple sclerosis reveals disease and c-Jun N-terminal kinase pathway biomarkers
Authors: Ferrandi Chiara , Richard Fabien , Tavano Patrizia , Hauben Ehud , Barbié Valerie , Gotteland Jean-Pierre , Greco Beatrice , Fortunato Mara , Mariani Maurizio F , Furlan Roberto , Comi Giancarlo , Martino Gianvito , Zaratin Paola F
Published: September 2010

Background:Autoimmune activation and deregulated apoptosis of T lymphocytes are involved in multiple sclerosis (MS). c-Jun N-terminal kinase (JNK) plays a role in T-cell survival and apoptosis.

Objectives:The aim of this work was to investigate the role of the JNK-dependent apoptosis pathway in relapsing–remitting MS (RRMS).

Methods:The immunomodulatory effect of AS602801, a JNK inhibitor, was firstly evaluated on activated peripheral blood mononuclear cells (PBMCs) from healthy volunteers (HVs) and secondly in unstimulated purified CD4+, CD8+ and CD11b+ cells from RRMS patients and HVs. Moreover JNK/inflammation/apoptosis related genes were investigated in RRMS and HV samples.

Results:In activated PBMCs from HVs, we showed that AS602801 blocked T-lymphocyte proliferation and induced apoptosis. In RRMS CD4+ and CD8+ cells, AS602801 induced apoptosis genes and expression of surface markers, while in RRMS CD11b+ cells it induced expression of innate immunity receptors and co-stimulatory molecules. Untreated cells from RRMS active-phase patients significantly released interleukin-23 (IL-23) and interferon-gamma (IFN-γ) and expressed less apoptosis markers compared to the cells of HVs. Moreover, gene expression was significantly different in cells from RRMS active-phase patients vs. HVs. By comparing RRMS PBMCs in the active and stable phases, a specific genomic signature for RRMS was indentified. Additionally, CASP8AP2, CD36, ITGAL, NUMB, OLR1, PIAS-1, RNASEL, RTN4RL2 and THBS1 were identified for the first time as being associated to the active phase of RRMS.

Conclusions:The analysis of the JNK-dependent apoptosis pathway can provide biomarkers for activated lymphocytes in the active phase of RRMS and a gene expression signature for disease status. The reported results might be useful to stratify patients, thereby supporting the development of novel therapies.
Active commuting to school in children and adolescents: An opportunity to increase physical activity and fitness
Authors: Chillón Palma , Ortega Francisco B. , Ruiz Jonatan R. , Veidebaum Toomas , Oja Leila , Mäestu Jarek , Sjöström Michael
Published: September 2010

Aims: The purpose was to describe the patterns of commuting to school in young people and to examine its associations with physical activity (PA) and cardiorespiratory fitness. Methods: The sample comprised 2271 Estonian and Swedish children and adolescents (1218 females) aged 9—10 years and 15—16 years. Data were collected in 1998/99. Mode of commuting to and from school was assessed by questionnaire. Time spent (min/day) in PA and average PA (counts/min) was measured by accelerometry. Cardiorespiratory fitness was assessed by means of a maximal cycle ergometer test. Results: Sixty-one percent of the participants reported active commuting to school (ACS). Estonian youth showed lower levels of ACS than Swedish (odds ratio, 0.64; 95% confidence interval, 0.53—0.76) and girls reported lower levels than boys (0.74; 0.62—0.88). ACS boys showed higher PA levels than non-ACS boys for moderate, vigorous, MVPA, and average PA levels (all p ≤ 0.01). Participants who cycled to school had higher cardiorespiratory fitness than walkers or passive travellers (p < 0.001). Conclusions: Nearly two-thirds of the participants actively commuted to school. ACS may provide an opportunity to increase levels of daily PA, especially in boys, and cardiorespiratory fitness, especially if cycling. Public health strategies should develop and test ACS patterns to get more evidence and promote bike-friendly environments.
The medium is the joke: online humor about and by networked computers
Authors: Shifman Limor , Blondheim Menahem
Published: September 2010

This article explores the uncharted territory of reflexive internet humor about networked computers. A combined quantitative—qualitative analysis of 250 texts sampled from popular websites yielded a map of the main themes underpinning this massive corpus of humor. We analyzed them in relation to three grand theories of the nature of humor — superiority, release, and incongruity — locating each theme on a matrix deriving from the theories: (i) a superiority axis, running between the powerful and weak players in the networked environment; (ii) an incongruity axis, running from the purely human to the strictly technical, and (iii) a release axis reflecting degrees of tension generated by the former two dualities. Our analysis suggests that humor about networked computers extends to a comment on the nature of humanness in a bewildering age of artificial intelligence. The communication of this reflexive comment may be shaping a global community of computer users.
The relationship between meniscal tears and meniscal position
Authors: Allen Deborah M. , Ling Li  , Crema Michel D. , Marra Monica D. , Guermazi Ali , Wyman Brad T. , Hellio Le Graverand Marie-Pierre , Englund Martin , Brandt Kenneth D. , Hunter David J.
Published: September 2010

Objective: To investigate how different types of meniscal tears predispose to different patterns of meniscal position in subjects with and without symptomatic knee osteoarthritis (OA).

Methods: A cross-sectional analysis of 161 women participating in an observational study to evaluate knee OA progression was performed using baseline MRI data. Meniscal morphologic features were scored in three separate locations. Meniscal position measures were determined for extrusion and proportion of coverage. Analysis was performed using multiple linear regression models treating each tear type as an individual variable with a binary response.

Results: Complex tears, cysts and maceration of the medial meniscus were associated with more medial ( p = 0.0004, p = 0.004, p < 0.0001, respectively) and anterior extrusion ( p = 0.03, p = 0.03, p < 0.0001, respectively) than normal menisci. Horizontal tears of the lateral meniscus had more lateral ( p = 0.005) and anterior extrusion ( p < 0.0001) than normal menisci. Anterior and body tears of the medial meniscus were associated with more anterior extrusion ( p = 0.0006, p = 0.01, respectively), whereas meniscal body tears alone had more medial extrusion than normal menisci ( p = 0.0002). Meniscal body tears of the lateral meniscus had more lateral extrusion than normal menisci ( p = 0.01).

Conclusion: Anterior horn and meniscal body tears and the more severe macerated and complex tear types predisposed to more medial meniscal extrusion. Laterally, only meniscal body and horizontal tears significantly affected extrusion, potentially reflecting a lower overall prevalence of lateral meniscal tears. These results may have important implications in identifying tear types associated with more meniscal dysfunction, with the ultimate goal of identifying those at greatest risk for knee OA progression.
Diminished contact-dependent reinforcement of Syk activation underlies impaired thrombus growth in mice lacking Semaphorin 4D
Authors: Wannemacher Kenneth M. , Zhu Li , Jiang Hong , Fong Karen P. , Stalker Timothy J. , Lee Dooyoung , Tran Anh N. , Neeves Keith B. , Maloney Sean , Kumanogoh Atsushi , Kikutani Hitoshi , Hammer Daniel A. , Diamond Scott L. , Brass Lawrence F.
Published: September 2010

We recently reported that Semaphorin 4D (Sema4D) and its receptors are expressed on the platelet surface and showed that Sema4D(−/−) mice have a selective defect in collagen-induced platelet aggregation and an impaired vascular injury response. Here we investigated the mechanisms involved, tested the role of platelet-platelet contacts in Sema4D-mediated events, and examined the relationship between Sema4D-dependent signaling and integrin αIIbβ3 outside-in signaling. The results show that spleen tyrosine kinase (Syk) activation, an early step in collagen signaling via the glycoprotein VI (GPVI)/FcRγ complex, is greatly reduced in Sema4D(−/−) platelets and can be restored by adding soluble Sema4D. Earlier events, including FcRγ phosphorylation, occur normally; later events are impaired. In contrast, when engagement of αIIbβ3 was blocked, Sema4D(−/−) and control platelets were indistinguishable in assays of Syk activation, adhesion, spreading on collagen, and activation of αIIbβ3. Finally, we found that, unlike the Sema4D knockout, αIIbβ3 blockade inhibited FcRγ phosphorylation and that stimulating aggregation with Mn2+ failed to normalize Syk activation in the absence of Sema4D. Collectively, these results show that αIIbβ3 and Sema4D jointly promote collagen responses by amplifying Syk activation, partly by forming integrin-mediated contacts that enable the binding of Sema4D to its receptors and partly through integrin outside-in signaling. These 2 processes are interdependent, but distinguishable.
Beyond the enhanceosome: cluster of novel κB sites downstream of the human IFN-β gene is essential for lipopolysaccharide-induced gene activation
Authors: Goh Fui G. , Thomson Scott J. P. , Krausgruber Thomas , Lanfrancotti Alessandra , Copley Richard R. , Udalova Irina A.
Published: September 2010

The expression of interferon-β (IFN-β) in virus-infected HeLa cells established a paradigm of multifactorial gene regulation, in which cooperative assembly of transcription factors (TFs) at the composite DNA element (enhanceosome), is central for amplification of weak activating signals provided by individual TFs. However, whether the same TFs and the same DNA element are essential for IFN-β induction in response to bacterial stimuli are less well understood. Here we report that rapid and transient transcription of IFN-β in response to TLR4 stimulation with bacterial lipopolysaccharide (LPS) follows nuclear factor-κB (NF-κB) RelA activation and recruitment to the IFN-β genomic locus at multiple spatially separated regulatory regions. We demonstrate that the IFN-β enhanceosome region is not sufficient for maximal gene induction in response to LPS and identify an essential cluster of homotypic κB sites in the 3′ downstream of the gene. The cluster is characterized by elevated levels of histone 3 lysine 4 mono-methylation, a chromatin signature of enhancers, and efficiently binds RelA-containing NF-κB complexes in vitro and in vivo. These findings demonstrate that IFN-β gene activation via multifactorial enhanceosome assembly is potentiated in LPS-stimulated cells by NF-κB interactions with all functional κB sites in the locus.
*omeSOM: a software for clustering and visualization of transcriptional and metabolite data mined from interspecific crosses of crop plants
Authors: Milone Diego H , Stegmayer Georgina S , Kamenetzky Laura , López Mariana , Lee Je Min , Giovannoni James J , Carrari Fernando
Published: September 2010

Background

Modern biology uses experimental systems that involve the exploration of phenotypic variation as a result of the recombination of several genomes. Such systems are useful to investigate the functional evolution of metabolic networks. One such approach is the analysis of transcript and metabolite profiles. These kinds of studies generate a large amount of data, which require dedicated computational tools for their analysis.

Results

This paper presents a novel software named *omeSOM (transcript/metabol-ome Self Organizing Map) that implements a neural model for biological data clustering and visualization. It allows the discovery of relationships between changes in transcripts and metabolites of crop plants harboring introgressed exotic alleles and furthermore, its use can be extended to other type of omics data. The software is focused on the easy identification of groups including different molecular entities, independently of the number of clusters formed. The *omeSOM software provides easy-to-visualize interfaces for the identification of coordinated variations in the co-expressed genes and co-accumulated metabolites. Additionally, this information is linked to the most widely used gene annotation and metabolic pathway databases.

Conclusions

*omeSOM is a software designed to give support to the data mining task of metabolic and transcriptional datasets derived from different databases. It provides a user-friendly interface and offers several visualization features, easy to understand by non-expert users. Therefore, *omeSOM provides support for data mining tasks and it is applicable to basic research as well as applied breeding programs. The software and a sample dataset are available free of charge at http://sourcesinc.sourceforge.net/omesom/.
Spindle assembly checkpoint genes reveal distinct as well as overlapping expression that implicates MDF-2/Mad2 in postembryonic seam cell proliferation in Caenorhabditis elegans
Authors: Tarailo-Graovac Maja , Wang Jun , Chu Jeffrey SC , Tu Domena , Baillie David L , Chen Nansheng
Published: September 2010

Background

The spindle assembly checkpoint (SAC) delays anaphase onset by inhibiting the activity of the anaphase promoting complex/cyclosome (APC/C) until all of the kinetochores have properly attached to the spindle. The importance of SAC genes for genome stability is well established; however, the roles these genes play, during postembryonic development of a multicellular organism, remain largely unexplored.

Results

We have used GFP fusions of 5' upstream intergenic regulatory sequences to assay spatiotemporal expression patterns of eight conserved genes implicated in the spindle assembly checkpoint function in Caenorhabditis elegans. We have shown that regulatory sequences for all of the SAC genes drive ubiquitous GFP expression during early embryonic development. However, postembryonic spatial analysis revealed distinct, tissue-specific expression of SAC genes with striking co-expression in seam cells, as well as in the gut. Additionally, we show that the absence of MDF-2/Mad2 (one of the checkpoint genes) leads to aberrant number and alignment of seam cell nuclei, defects mainly attributed to abnormal postembryonic cell proliferation. Furthermore, we show that these defects are completely rescued by fzy-1(h1983)/CDC20, suggesting that regulation of the APC/CCDC20 by the SAC component MDF-2 is important for proper postembryonic cell proliferation.

Conclusion

Our results indicate that SAC genes display different tissue-specific expression patterns during postembryonic development in C. elegans with significant co-expression in hypodermal seam cells and gut cells, suggesting that these genes have distinct as well as overlapping roles in postembryonic development that may or may not be related to their established roles in mitosis. Furthermore, we provide evidence, by monitoring seam cell lineage, that one of the checkpoint genes is required for proper postembryonic cell proliferation. Importantly, our research provides the first evidence that postembryonic cell division is more sensitive to SAC loss, in particular MDF-2 loss, than embryonic cell division.
EMG1 is essential for mouse pre-implantation embryo development
Authors: Wu Xiaoli , Sandhu Sumit , Patel Nehal , Triggs-Raine Barbara , Ding Hao
Published: September 2010

Background

Essential for mitotic growth 1 (EMG1) is a highly conserved nucleolar protein identified in yeast to have a critical function in ribosome biogenesis. A mutation in the human EMG1 homolog causes Bowen-Conradi syndrome (BCS), a developmental disorder characterized by severe growth failure and psychomotor retardation leading to death in early childhood. To begin to understand the role of EMG1 in mammalian development, and how its deficiency could lead to Bowen-Conradi syndrome, we have used mouse as a model. The expression of Emg1 during mouse development was examined and mice carrying a null mutation for Emg1 were generated and characterized.

Results

Our studies indicated that Emg1 is broadly expressed during early mouse embryonic development. However, in late embryonic stages and during postnatal development, Emg1 exhibited specific expression patterns. To assess a developmental role for EMG1 in vivo, we exploited a mouse gene-targeting approach. Loss of EMG1 function in mice arrested embryonic development prior to the blastocyst stage. The arrested Emg1

-/-
embryos exhibited defects in early cell lineage-specification as well as in nucleologenesis. Further, loss of p53, which has been shown to rescue some phenotypes resulting from defects in ribosome biogenesis, failed to rescue the Emg1

-/-
pre-implantation lethality.

Conclusion

Our data demonstrate that Emg1 is highly expressed during mouse embryonic development, and essential for mouse pre-implantation development. The absolute requirement for EMG1 in early embryonic development is consistent with its essential role in yeast. Further, our findings also lend support to the previous study that showed Bowen-Conradi syndrome results from a partial EMG1 deficiency. A complete deficiency would not be expected to be compatible with a live birth.
The blue lizard spandrel and the island syndrome
Authors: Raia Pasquale , Guarino Fabio M , Turano Mimmo , Polese Gianluca , Rippa Daniela , Carotenuto Francesco , Monti Daria M , Cardi Manuela , Fulgione Domenico
Published: September 2010

Background

Many small vertebrates on islands grow larger, mature later, lay smaller clutches/litters, and are less sexually dimorphic and aggressive than their mainland relatives. This set of observations is referred to as the 'Island Syndrome'. The syndrome is linked to high population density on islands. We predicted that when population density is low and/or fluctuating insular vertebrates may evolve correlated trait shifts running opposite to the Island Syndrome, which we collectively refer to as the 'reversed island syndrome' (RIS) hypothesis. On the proximate level, we hypothesized that RIS is caused by increased activity levels in melanocortin receptors. Melanocortins are postranslational products of the proopiomelanocortin gene, which controls pleiotropically pigmentation, aggressiveness, sexual activity, and food intake in vertebrates.

Results

We tested the RIS hypothesis performing a number of behavioral, genetic, and ontogenetic tests on a blue colored insular variant of the Italian Wall lizardPodarcis sicula, living on a small island off the Southern Italian coast. The population density of this blue-colored variant was generally low and highly fluctuating from one year to the next.

In keeping with our predictions, insular lizards were more aggressive and sexually dimorphic than their mainland relatives. Insular males had wide, peramorphic heads. The growth rate of insular females was slower than growth rates of mainland individuals of both sexes, and of insular males. Consequently, size and shape dimorphism are higher on the Island. As predicted, melanocortin receptors were much more active in individuals of the insular population. Insular lizards have a higher food intake rate than mainland individuals, which is consistent with the increased activity of melanocortin receptors. This may be adaptive in an unpredictable environment such as Licosa Island. Insular lizards of both sexes spent less time basking than their mainland relatives. We suspect this is a by-product (spandrel) of the positive selection for increased activity of melanocortins receptors.

Conclusions

We contend that when population density is either low or fluctuating annually as a result of environmental unpredictability, it may be advantageous to individuals to behave more aggressively, to raise their rate of food intake, and allocate more energy into reproduction.
Bystander T cells in human immune responses to dengue antigens
Authors: Suwannasaen Duangchan , Romphruk Arunrat , Leelayuwat Chanvit , Lertmemongkolchai Ganjana
Published: September 2010

Background

Previous studies of T cell activation in dengue infection have focused on restriction of specific T cell receptors (TCRs) and classical MHC molecules. However, bystander T cell activation, which is TCR independent, occurs via cytokines in other viral infections, both in vitro and in vivo, and enables T cells to bypass certain control checkpoints. Moreover, clinical and pathological evidence has pointed to cytokines as the mediators of dengue disease severity. Therefore, we investigated bystander T cell induction by dengue viral antigen.

Results

Whole blood samples from 55 Thai schoolchildren aged 13-14 years were assayed for in vitro interferon-gamma (IFN-γ) induction in response to inactivated dengue serotype 2 antigen (Den2). The contribution of TCR-dependent and independent pathways was tested by treatment with cyclosporin A (CsA), which inhibits TCR-dependent activation of T cells. ELISA results revealed that approximately 72% of IFN-γ production occurred via the TCR-dependent pathway. The major IFN-γ sources were natural killer (NK) (mean ± SE = 55.2 ± 3.3), CD4+T (24.5 ± 3.3) and CD8+T cells (17.9 ± 1.5), respectively, as demonstrated by four-color flow cytometry. Interestingly, in addition to these cells, we found CsA-resistant IFN-γ producing T cells (CD4+T = 26.9 ± 3.6% and CD8+T = 20.3 ± 2.1%) implying the existence of activated bystander T cells in response to dengue antigen in vitro. These bystander CD4+and CD8+T cells had similar kinetics to NK cells, appeared after 12 h and were inhibited by anti-IL-12 neutralization indicating cytokine involvement.

Conclusions

This study described immune cell profiles and highlighted bystander T cell activation in response to dengue viral antigens of healthy people in an endemic area. Further studies on bystander T cell activation in dengue viral infection may reveal the immune mechanisms that protect or enhance pathogenesis of secondary dengue infection.
Palliative care for the elderly - developing a curriculum for nursing and medical students
Authors: Just Johannes M , Schulz Christian , Bongartz Maren , Schnell Martin W
Published: September 2010

Background

Delivering palliative care to elderly, dying patients is a present and future challenge. In Germany, this has been underlined by a 2009 legislation implementing palliative care as compulsory in the medical curriculum. While the number of elderly patients is increasing in many western countries multimorbidity, dementia and frailty complicate care. Teaching palliative care of the elderly to an interprofessional group of medical and nursing students can help to provide better care as acknowledged by the ministry of health and its expert panels.

In this study we researched and created an interdisciplinary curriculum focussing on the palliative care needs of the elderly which will be presented in this paper.

Methods

In order to identify relevant learning goals and objectives for the curriculum, we proceeded in four subsequent stages.

We searched international literature for existing undergraduate palliative care curricula focussing on the palliative care situation of elderly patients; we searched international literature for palliative care needs of the elderly. The searches were sensitive and limited in nature. Mesh terms were used where applicable. We then presented the results to a group of geriatrics and palliative care experts for critical appraisal. Finally, the findings were transformed into a curriculum, focussing on learning goals, using the literature found.

Results

The literature searches and expert feedback produced a primary body of results. The following deduction domains emerged: Geriatrics, Palliative Care, Communication & Patient Autonomy and Organisation & Social Networks. Based on these domains we developed our curriculum.

Conclusions

The curriculum was successfully implemented following the Kern approach for medical curricula. The process is documented in this paper. The information given may support curriculum developers in their search for learning goals and objectives.
Manipulative therapy in addition to usual medical care accelerates recovery of shoulder complaints at higher costs: economic outcomes of a randomized trial
Authors: Bergman Gert JD , Winter Jan C , van Tulder Maurits W , Meyboom-de Jong Betty , Postema Klaas , van der Heijden Geert JMG
Published: September 2010

Background

Shoulder complaints are common in primary care and have unfavourable long term prognosis. Our objective was to evaluate the clinical effectiveness of manipulative therapy of the cervicothoracic spine and the adjacent ribs in addition to usual medical care (UMC) by the general practitioner in the treatment of shoulder complaints.

Methods

This economic evaluation was conducted alongside a randomized trial in primary care. Included were 150 patients with shoulder complaints and a dysfunction of the cervicothoracic spine and adjacent ribs. Patients were treated with UMC (NSAID's, corticosteroid injection or referral to physical therapy) and were allocated at random (yes/no) to manipulative therapy (manipulation and mobilization). Patient perceived recovery, severity of main complaint, shoulder pain, disability and general health were outcome measures. Data about direct and indirect costs were collected by means of a cost diary.

Results

Manipulative therapy as add-on to UMC accelerated recovery on all outcome measures included. At 26 weeks after randomization, both groups reported similar recovery rates (41% vs. 38%), but the difference between groups in improvement of severity of the main complaint, shoulder pain and disability sustained. Compared to the UMC group the total costs were higher in the manipulative group (€1167 vs. €555). This is explained mainly by the costs of the manipulative therapy itself and the higher costs due sick leave from work. The cost effectiveness ratio showed that additional manipulative treatment is more costly but also more effective than UMC alone. The cost-effectiveness acceptability curve shows that a 50%-probability of recovery with AMT within 6 months after initiation of treatment is achieved at €2876.

Conclusion

Manipulative therapy in addition to UMC accelerates recovery and is more effective than UMC alone on the long term, but is associated with higher costs.

International Standard Randomized Controlled Trial Number Register

ISRCTN11216
Evaluation of combinatorial cis-regulatory elements for stable gene expression in chicken cells
Authors: Seo Hee W , Kim Tae M , Choi Jin W , Han Beom K , Song Gwonhwa , Han Jae Y
Published: September 2010

Background

Recent successes in biotechnological application of birds are based on their unique physiological traits such as unlimited manipulability onto developing embryos and simple protein constituents of the eggs. However it is not likely that target protein is produced as kinetically expected because various factors affect target gene expression. Although there have been various attempts to minimize the silencing of transgenes, a generalized study that uses multiple cis-acting elements in chicken has not been made. The aim of the present study was to analyze whether various cis-acting elements can help to sustain transgene expression in chicken fibroblasts.

Results

We investigated the optimal transcriptional regulatory elements for enhancing stable transgene expression in chicken cells. We generated eight constructs that encode enhanced green fluorescent protein (eGFP) driven by either CMV or CAG promoters (including the control), containing three types of key regulatory elements: a chicken lysozyme matrix attachment region (cMAR), 5'-DNase I-hypersensitive sites 4 (cHS4), and the woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Then we transformed immortalized chicken embryonic fibroblasts with these constructs by electroporation, and after cells were expanded under G418 selection, analyzed mRNA levels and mean fluorescence intensity (MFI) by quantitative real-time PCR and flow cytometry, respectively. We found that the copy number of each construct significantly decreased as the size of the construct increased (R2 = 0.701). A significant model effect was found in the expression level among various constructs in both mRNA and protein (P < 0.0001). Transcription with the CAG promoter was 1.6-fold higher than the CMV promoter (P = 0.027) and the level of eGFP expression activity in cMAR- or cHS4-flanked constructs increased by two- to three-fold compared to the control CMV or CAG promoter constructs. In addition, flow cytometry analysis showed that constructs having cis-acting elements decreased the level of gene silencing as well as the coefficient of variance of eGFP-expressing cells (P < 0.0001).

Conclusions

Our current data show that an optimal combination of cis-acting elements and promoters/enhancers for sustaining gene expression in chicken cells is suggested. These results provide important information for avian transgenesis and gene function studies in poultry.
Fed-batch process for the psychrotolerant marine bacterium Pseudoalteromonas haloplanktis
Authors: Wilmes Boris , Hartung Angelika , Lalk Michael , Liebeke Manuel , Schweder Thomas , Neubauer Peter
Published: September 2010

Background

Pseudoalteromonas haloplanktis is a cold-adapted γ-proteobacterium isolated from Antarctic sea ice. It is characterized by remarkably high growth rates at low temperatures. P. haloplanktis is one of the model organisms of cold-adapted bacteria and has been suggested as an alternative host for the soluble overproduction of heterologous proteins which tend to form inclusion bodies in established expression hosts. Despite the progress in establishing P. haloplanktis as an alternative expression host the cell densities obtained with this organism, which is unable to use glucose as a carbon source, are still low. Here we present the first fed-batch cultivation strategy for this auspicious alternative expression host.

Results

The key for the fed-batch cultivation of P. haloplanktis was the replacement of peptone by casamino acids, which have a much higher solubility and allow a better growth control. In contrast to the peptone medium, on which P. haloplanktis showed different growth phases, on a casamino acids-containing, phosphate-buffered medium P. haloplanktis grew exponentially with a constant growth rate until the stationary phase. A fed-batch process was established by feeding of casamino acids with a constant rate resulting in a cell dry weight of about 11 g l-1 (OD540 = 28) which is a twofold increase of the highest densities which have been obtained with P. haloplanktis so far and an eightfold increase of the density obtained in standard shake flask cultures.

The cell density was limited in the fed-batch cultivation by the relatively low solubility of casamino acids (about 100 g l-1), which was proven by pulse addition of casamino acid powder which increased the cell density to about 20 g l-1 (OD540 = 55).

Conclusion

The growth of P. haloplanktis to higher cell densities on complex medium is possible. A first fed-batch fermentation strategy could be established which is feasible to be used in lab-scale or for industrial purposes. The substrate concentration of the feeding solution was found to influence the maximal biomass yield considerably. The bottleneck for growing P. haloplanktis to high cell densities still remains the availability of a highly concentrated substrate and the reduction of the substrate complexity. However, our results indicate glutamic acid as a major carbon source, which provides a good basis for further improvement of the fed-batch process.
A large proportion of asymptomatic Plasmodium infections with low and sub-microscopic parasite densities in the low transmission setting of Temotu Province, Solomon Islands: challenges for malaria diagnostics in an elimination setting
Authors: Harris Ivor , Sharrock Wesley W , Bain Lisa M , Gray Karen-Ann , Bobogare Albino , Boaz Leonard , Lilley Ken , Krause Darren , Vallely Andrew , Johnson Marie-Louise , Gatton Michelle L , Shanks G Dennis , Cheng Qin
Published: September 2010

Background

Many countries are scaling up malaria interventions towards elimination. This transition changes demands on malaria diagnostics from diagnosing ill patients to detecting parasites in all carriers including asymptomatic infections and infections with low parasite densities. Detection methods suitable to local malaria epidemiology must be selected prior to transitioning a malaria control programme to elimination. A baseline malaria survey conducted in Temotu Province, Solomon Islands in late 2008, as the first step in a provincial malaria elimination programme, provided malaria epidemiology data and an opportunity to assess how well different diagnostic methods performed in this setting.

Methods

During the survey, 9,491 blood samples were collected and examined by microscopy for Plasmodium species and density, with a subset also examined by polymerase chain reaction (PCR) and rapid diagnostic tests (RDTs). The performances of these diagnostic methods were compared.

Results

A total of 256 samples were positive by microscopy, giving a point prevalence of 2.7%. The species distribution was 17.5% Plasmodium falciparum and 82.4% Plasmodium vivax. In this low transmission setting, only 17.8% of the P. falciparum and 2.9% of P. vivax infected subjects were febrile (≥38°C) at the time of the survey. A significant proportion of infections detected by microscopy, 40% and 65.6% for P. falciparum and P. vivax respectively, had parasite density below 100/μL. There was an age correlation for the proportion of parasite density below 100/μL for P. vivax infections, but not for P. falciparum infections. PCR detected substantially more infections than microscopy (point prevalence of 8.71%), indicating a large number of subjects had sub-microscopic parasitemia. The concordance between PCR and microscopy in detecting single species was greater for P. vivax (135/162) compared to P. falciparum (36/118). The malaria RDT detected the 12 microscopy and PCR positive P. falciparum, but failed to detect 12/13 microscopy and PCR positive P. vivax infections.

Conclusion

Asymptomatic malaria infections and infections with low and sub-microscopic parasite densities are highly prevalent in Temotu province where malaria transmission is low. This presents a challenge for elimination since the large proportion of the parasite reservoir will not be detected by standard active and passive case detection. Therefore effective mass screening and treatment campaigns will most likely need more sensitive assays such as a field deployable molecular based assay.
A proteomic view of Caenorhabditis elegans caused by short-term hypoxic stress
Authors: Li Hualing , Ren Changhong , Shi Jinping , Hang Xingyi , Zhang Feilong , Gao Yan , Wu Yonghong , Xu Langlai , Chen Changsheng , Zhang Chenggang
Published: September 2010

Background

The nematode Caenorhabditis elegans is both sensitive and tolerant to hypoxic stress, particularly when the evolutionarily conserved hypoxia response pathway HIF-1/EGL-9/VHL is involved. Hypoxia-induced changes in the expression of a number of genes have been analyzed using whole genome microarrays in C. elegans, but the changes at the protein level in response to hypoxic stress still remain unclear.

Results

Here, we utilized a quantitative proteomic approach to evaluate changes in the expression patterns of proteins during the early response to hypoxia in C. elegans. Two-dimensional difference gel electrophoresis (2D-DIGE) was used to compare the proteomic maps of wild type C. elegans strain N2 under a 4-h hypoxia treatment (0.2% oxygen) and under normoxia (control). A subsequent analysis by MALDI-TOF-TOF-MS revealed nineteen protein spots that were differentially expressed. Nine of the protein spots were significantly upregulated, and ten were downregulated upon hypoxic stress. Three of the upregulated proteins were involved in cytoskeletal function (LEV-11, MLC-1, ACT-4), while another three upregulated (ATP-2, ATP-5, VHA-8) were ATP synthases functionally related to energy metabolism. Four ribosomal proteins (RPL-7, RPL-8, RPL-21, RPS-8) were downregulated, indicating a decrease in the level of protein translation upon hypoxic stress. The overexpression of tropomyosin (LEV-11) was further validated by Western blot. In addition, the mutant strain of lev-11(x12) also showed a hypoxia-sensitive phenotype in subsequent analyses, confirming the proteomic findings.

Conclusions

Taken together, our data suggest that altered protein expression, structural protein remodeling, and the reduction of translation might play important roles in the early response to oxygen deprivation in C. elegans, and this information will help broaden our knowledge on the mechanism of hypoxia response.
Overestimation of physical activity level is associated with lower BMI: a cross-sectional analysis
Authors: Watkinson Clare , van Sluijs Esther MF , Sutton Stephen , Hardeman Wendy , Corder Kirsten , Griffin Simon J
Published: September 2010

Background

Poor recognition of physical inactivity may be an important barrier to healthy behaviour change, but little is known about this phenomenon. We aimed to characterize a high-risk population according to the discrepancies between objective and self-rated physical activity (PA), defined as awareness.

Methods

An exploratory cross-sectional analysis of PA awareness using baseline data collected from 365 ProActive participants between 2001 and 2003 in East Anglia, England. Self-rated PA was defined as 'active' or 'inactive' (assessed via questionnaire). Objective PA was defined according to achievement of guideline activity levels (≥30 minutes or <30 minutes spent at least moderate intensity PA, assessed by heart rate monitoring). Four awareness groups were created: 'Realistic Actives', 'Realistic Inactives', 'Overestimators' and 'Underestimators'. Logistic regression was used to assess associations between awareness group and 17 personal, social and biological correlates.

Results

63.3% of participants (N = 231) were inactive according to objective measurement. Of these, 45.9% rated themselves as active ('Overestimators'). In a multiple logistic regression model adjusted for age and smoking, males (OR = 2.11, 95% CI = 1.12, 3.98), those with lower BMI (OR = 0.89, 95% CI = 0.84, 0.95), younger age at completion of full-time education (OR = 0.83, 95% CI = 0.74, 0.93) and higher general health perception (OR = 1.02 CI = 1.00, 1.04) were more likely to overestimate their PA.

Conclusions

Overestimation of PA is associated with favourable indicators of relative slimness and general health. Feedback about PA levels could help reverse misperceptions.