bluuebunny/crossref_metadata_2025 huggingface dataset:
Patterns of Medicaid Disenrollment for Youth With Mental Health Problems
Authors: Pullmann Michael D. , Heflinger Craig Anne , Satterwhite Mayberry Lindsay
Published: June 2010

Young people who receive Medicaid-funded mental health services during the transition to adulthood often face disenrollment from Medicaid without continuity into publicly funded services. This article investigates the longitudinal predictors of these coverage gaps and disenrollment from age 16 to 23 years. Cox regression analyses estimated predictors of time until the first loss of coverage for 180 days or more, and time until final disenrollment with no subsequent reenrollment. Females were much more likely to regain and retain coverage after initial loss. Funding source and diagnoses predicted Medicaid retention differentially by gender. For both genders, funding through Social Security Income or a diagnosis of Mental Retardation/ Developmental Disabilities was related to Medicaid retention. Disenrollment especially affected males precisely at their 18th and 19th birthdays. Nearly one third of females qualified for Medicaid because of pregnancy. Eligibility guidelines relate to retention and loss during the transition to adulthood and may need reevaluation to ensure continuity of care.
Health Care Delivery Restructuring and Productivity Change: Assessing the Veterans Integrated Service Networks (VISNs) Using the Malmquist Approach
Authors: Ozcan Yasar A. , Luke Roice D.
Published: June 2010

This study evaluates the productivity changes for the Veterans Integrated Service Networks (VISNs) that the Veterans Health Administration (VHA) created, comparing performance in 1994 with that in 2004. This represents periods before and after the VHA in 1995 reconfigured provider units into 21 regionalized delivery systems and engaged in other important system innovations. Productivity is measured using the Malmquist Index approach (a longitudinal version of the data envelopment analysis [DEA]). Results indicate that the VISN restructuring generally produced improvements in overall productivity (Malmquist scores) and in VISN adaptations to structural/technological change. They also show that the VISNs overall did not produce “changes in efficiency,” reflecting challenges they may have faced in making “technical change” through management adaptations. The findings are consistent with what would be expected, given the major changes that did occur within the VHA in recent years as well as the before and after design used in this study.
Missing Productivity Gains in the Medicare Physician Fee Schedule: Where Are They?
Authors: Cromwell Jerry , McCall Nancy , Dalton Kathleen , Braun Peter
Published: June 2010

The Medicare Fee Schedule with payments for thousands of visits and procedures is updated periodically for the work component of changes in physician relative work. Three 5-year reviews of physician work by Medicare have been biased against finding productivity gains and reductions in physician work relative values. The authors present four studies showing shorter physician times with patients in their offices and in the operating room, increases in surgeons’ self-reported total work in spite of declining operating room times, and growing numbers of costly handoffs to nonsurgeons, while surgeons receive full payment for postoperative follow-up with patients. Substantial savings exist in the fee schedule if productivity gains from greater delegation to ancillary staff and specialists, reengineering of services, and rapid learning by experience with new technologies were integrated into the periodic reviews.
Defining and Measuring Integrated Patient Care: Promoting the Next Frontier in Health Care Delivery
Authors: Singer Sara J. , Burgers Jako , Friedberg Mark , Rosenthal Meredith B. , Leape Lucian , Schneider Eric
Published: June 2010

Integration of care is emerging as a central challenge of health care delivery, particularly for patients with multiple, complex chronic conditions. The authors argue that the concept of “integrated patient care” would benefit from further clarification regarding (a) the object of integration and (b) its essential components, particularly when constructing measures.To address these issues, the authors propose a definition of integrated patient care that distinguishes it from integrated delivery organizations, acknowledging that integrated organizational structures and processes may fail to produce integrated patient care. The definition emphasizes patients’ central role as active participants in managing their own health by including patient centeredness as a key element of integrated patient care. Measures based on the proposed definition will enable empirical assessment of the potential relationships between the integration of organizations, the integration of patient care, and patient outcomes, providing valuable guidance to health systems reformers.
Making Sense of Place: Mapping as a Multisensory Research Method
Authors: Powell Kimberly
Published: June 2010

This article argues for the importance of mapping as a multisensory research method in terms of its ability to evoke relationships between place, lived experience, and community. Based on an interdisciplinary summer research course for graduate and undergraduate students that focused on the analysis and design of appropriate development strategies for the El Chorrillo neighborhood in Panama City, Panama, the author describes a research project that combined arts-based research, design and urban planning methods, and ethnography to develop visual fieldwork methods for site-based research, urban planning, and community development in which mapping featured prominently as part of the research. The purpose of this article is to establish an argument for the unique contribution of mapping as a qualitative method, particularly in the ways that contemporary aesthetics of mapping can be used to evoke the lived experience of social, cultural, and political issues related to place.
The Hermit and the Old Goat
Authors: Giorgio Grace
Published: June 2010

Narrated by a man looking back at his childhood, The Hermit and the Old Goat is loosely connected to real events. Placing herself in this fictional character’s mind allowed the author to explore with distant compassion the results of a traumatic family split. In self-narrative, we can re-script a traumatic experience’s meaning without having to depict it exactly as it happened. The trauma of losing one’s family disrupts one’s sense of self, one’s connection to the past and present and even the future. The Hermit and the Old Goat explores such trauma and its aftereffects from the reflective view of an outsider and his community. Such narrative distance in time, character, and even gender allows for an expression of forgiveness, understanding, and healing.
Layered Reflections on a Written Story Through Real-Time Conversation
Authors: Frentz Thomas S. , Hocker Joyce L.
Published: June 2010

In this work, the authors develop a dialogical method for generating layered reflections about a written story. The authors then apply this method to an existing narrative so as to discover important, but nonobvious meanings that might facilitate the psychological growth of the storyteller. The dialogical interchange used here to prompt deeper reflections will hopefully generalize to anyone using this method.
Association Between Intimate Partner Violence and Irritable Bowel Syndrome: A Population-Based Study in Nicaragua
Authors: Becker-Dreps Sylvia , Morgan Douglas , Peña Rodolfo , Cortes Loreto , Martin Christopher F. , Valladares Eliette
Published: June 2010

Irritable bowel syndrome (IBS) is a disabling functional gastrointestinal disorder, which serves as a model for abdominal pain syndromes. An association between intimate partner violence and IBS has been shown among White women in the industrialized world. To determine whether this relationship transcends cultural boundaries, we conducted a population-based, cross-sectional survey in Nicaragua using the innovative Health and Demographic Surveillance System in the León province. Women who had experienced physical intimate partner violence had significantly increased risk of IBS (odds ratio [OR] = 2.08; 95% confidence interval [CI] = 1.35, 3.21), as did those who had experienced sexual intimate partner violence (OR = 2.85; 95% CI = 1.45, 5.59). These findings argue for intimate partner violence screening among Latina women with IBS.
Affect Recognition in Adults With ADHD
Authors: Miller Meghan , Hanford Russell B. , Fassbender Catherine , Duke Marshall , Schweitzer Julie B.
Published: June 2010

Objective: This study compared affect recognition abilities between adults with and without ADHD. Method: The sample consisted of 51 participants (34 men, 17 women) divided into 3 groups: ADHD-combined type (ADHD-C; n = 17), ADHD-predominantly inattentive type (ADHD-I; n = 16), and controls ( n = 18). The mean age was 34 years. Affect recognition abilities were assessed by the Diagnostic Analysis of Nonverbal Accuracy (DANVA). Results: ANOVA showed that the ADHD-I group made more fearful emotion errors relative to the control group. Inattentive symptoms were positively correlated, whereas hyperactive-impulsive symptoms were negatively correlated with affect recognition errors. Conclusion: These results suggested that affect recognition abilities may be impaired in adults with ADHD and that affect recognition abilities are more adversely affected by inattentive than hyperactive-impulsive symptoms.
Two-year study of cervical cord volume and myelin water in primary progressive multiple sclerosis
Authors: Laule C. , Vavasour IM , Zhao Y. , Traboulsee AL , Oger J. , Vavasour JD , Mackay AL , Li DKB
Published: June 2010

Background: Spinal cord involvement in multiple sclerosis (MS) is common and an important element in disability. Previous studies demonstrated smaller cervical cord area at the C2 level in MS compared to controls, and a decrease in cord area over 12 months, most marked in primary progressive MS (PPMS). A subset of subjects participating in a multicentre, double-blind, placebo-controlled clinical trial evaluating the efficacy of glatiramer acetate in PPMS (PROMiSe trial) were followed for 2 years.

Methods: 24 PPMS subjects, randomized to placebo ( n = 9) and glatiramer acetate ( n = 15), and 24 matched controls were studied. Cervical cord volume (CCV) at C2—3 was determined using a 3D inversion recovery (IR)-prepared spoiled-gradient echo sequence. Myelin water fraction (MWF) at C2—3 was obtained using a 32-echo IR-prepared relaxation sequence. Scans were repeated at baseline, years 1 and 2.

Results: Baseline CCV was significantly smaller for PPMS than controls [median (interquartile range) 951 (829—1043) vs. 1072 (1040—1129) mm3, p = 0.0004] and MWF trended to be lower in PPMS cord [median (interquartile range) 0.225 (0.187—0.267) vs. 0.253 (0.235—0.266), p = 0.12]. Baseline CCV correlated with baseline Expanded Disability Status Scale, disease duration, brain white and grey matter volume. In PPMS, CCV was significantly decreased at year 1 (—0.83%, p = 0.04) and year 2 (—1.65%, p = 0.02). Baseline MWF correlated with baseline CCV and brain white and grey matter volume. MWF was significantly decreased from baseline for PPMS at year 2 (—10.5%, p = 0.01). Treatment effect was not detected on change in CCV nor MWF.

Conclusions: Metrics at the level of the cord, including volume and MWF at C2—3, were lower in PPMS than controls and changed over 2 years only in PPMS.
Realism and the spirit of 1919: Halford Mackinder, geopolitics and the reality of the League of Nations
Authors: Ashworth Lucian M.
Published: June 2010

Recent analyses of interwar International Relations (IR) have argued that there was no realist–idealist debate, and that there is no evidence of a distinct idealist paradigm. Less work has been done on realism in the interwar period. This article analyses the thought of one particular early 20th-century realist: Halford J. Mackinder. A product of the development of political geography, and a major influence on American strategic studies, Mackinder is best known for his Heartland thesis, which has been interpreted as environmental determinism. Yet, Mackinder’s realism is a complex mix of geopolitical analysis and the influence of ideas on human action. His concepts of organizer and idealist foreign policy ideal types pre-date Carr’s realist–utopian distinction by two decades, while his interpretation of the realities of international politics is at odds with Morgenthau’s realism. A closer analysis of Mackinder’s realism (1) underscores the links between geopolitics and realist strategic studies; (2) demonstrates the diversity of realist approaches in interwar IR; and (3) shows that it was possible to be a realist and also support the League of Nations. There are limits to Mackinder’s usefulness to 21st-century IR, but an understanding of his brand of realism is necessary for a fuller understanding of the development of realism as a 20th-century school of thought.
War memories and Japan’s ‘normalization’ as an international actor: A critical analysis
Authors: Lawson Stephanie , Tannaka Seiko
Published: June 2010

The paradigmatic case in which an almost permanent impasse exists in coming to terms with a difficult war past and ‘normalizing’ its international relations is that of Japan. Although successive Japanese governments have apologized over the last few decades, these have been countered by periodic episodes within Japan revolving largely around history textbooks, the remembrance of war dead and the quest by nationalists to restore national pride in the past. Regional relations were especially strained during the premiership of Koizumi Juni’chirô and his immediate successor, Abe Shinzô. They improved under Fukuda Yasuo, a moderate on war memory issues, and remained steady under Asô Tarô. Japan’s latest prime minister, Hatoyama Yukio, appears determined to address Japan’s war past more openly and critically than previous LDP figures, no doubt with an eye to improving Japan’s relations with its Asian neighbours. But whatever line he pursues, contestation over war memories will remain an issue. They are driven by deep divisions within Japan at the same time that political leaders seek a more prominent identity for Japan as a ‘normal’ actor in international affairs. This article analyses key aspects of the politics of Japan’s war memories, using insights from collective memory studies and constructivist IR theory. We suggest that the quest for ‘normality’ has generated a set of tensions and contradictions over the issue of war memories, both domestically and internationally, for which there is no resolution in sight.
Vegf regulates embryonic erythroid development through Gata1 modulation
Authors: Drogat Benjamin , Kalucka Joanna , Gutiérrez Laura , Hammad Hamida , Goossens Steven , Farhang Ghahremani Morvarid , Bartunkova Sonia , Haigh Katharina , Deswarte Kim , Nyabi Omar , Naessens Michael , Ferrara Napoleone , Klingmüller Ursula , Lambrecht Bart N. , Nagy Andras , Philipsen Sjaak , Haigh Jody J.
Published: June 2010

To determine the role of vascular endothelial growth factor (Vegf) in embryonic erythroid development we have deleted or overexpressed Vegf specifically in the erythroid lineage using the EpoR-iCre transgenic line in combination with Cre/loxP conditional gain and loss of function Vegf alleles. ROSA26 promoter-based expression of the Vegf164 isoform in the early erythroid lineage resulted in a differentiation block of primitive erythroid progenitor (EryP) development and a partial block in definitive erythropoiesis between the erythroid burst-forming unit and erythroid colony-forming unit stages. Decreased mRNA expression levels of the key erythroid transcription factor Gata1 were causally linked to this phenotype. Conditional deletion of Vegf within the erythroid lineage was associated with increased Gata1 levels and increased erythroid differentiation. Expression of a ROSA26-based GATA2 transgene rescued Gata1 mRNA levels and target genes and restored erythroid differentiation in our Vegf gain of function model. These results demonstrate that Vegf modulates Gata1 expression levels in vivo and provides new molecular insight into Vegf's ability to modulate erythropoiesis.
Genetic variation in hyaluronan metabolism loci is associated with plasma plasminogen activator inhibitor-1 concentration
Authors: Lanktree Matthew B. , Johansen Christopher T. , Anand Sonia S. , Davis A. Darlene , Miller Ruby , Yusuf Salim , Hegele Robert A. ,   on behalf of the SHARE, and SHARE-AP Investigators
Published: June 2010

Elevated plasma plasminogen activator inhibitor-1 (PAI-1) concentration is associated with cardiovascular disease risk. PAI-1 is the primary inhibitor of fibrinolysis within both the circulation and the arterial wall, playing roles in both atherosclerosis and thrombosis. To define the heritable component, subjects within the population-based SHARE (Study of Health Assessment and Risk in Ethnic groups) and SHARE-AP (Study of Health Assessment and Risk Evaluation in Aboriginal Peoples) studies, composed of Canadians of South Asian (n = 298), Chinese (n = 284), European (n = 227), and Aboriginal (n = 284) descent, were genotyped using the gene-centric Illumina HumanCVD BeadChip. After imputation, more than 150 000 single nucleotide polymorphisms (SNPs) in more than 2000 loci were tested for association with plasma PAI-1 concentration. Marginal association was observed with the PAI-1 locus itself (SERPINE1; P < .05). However, 5 loci (HABP2, HSPA1A, HYAL1, MBTPS1, TARP) were associated with PAI-1 concentration at a P < 1 × 10−5 threshold. The protein products of 2 of these loci, hyaluronan binding protein 2 (HABP2) and hyaluronoglucosaminidase 1 (HYAL1), play key roles in hyaluronan metabolism, providing genetic evidence to link these pathways.
Immobilizing topoisomerase I on a surface plasmon resonance biosensor chip to screen for inhibitors
Authors: Tsai Hsiang-Ping , Lin Li-Wei , Lai Zhi-Yang , Wu Jui-Yu , Chen Chiao-En , Hwang Jaulang , Chen Chien-Shu , Lin Chun-Mao
Published: June 2010

Background

The topoisomerase I (TopI) reaction intermediate consists of an enzyme covalently linked to a nicked DNA molecule, known as a TopI-DNA complex, that can be trapped by inhibitors and results in failure of re-ligation. Attempts at new derivative designs for TopI inhibition are enthusiastically being pursued, and TopI inhibitors were developed for a variety of applications. Surface plasmon resonance (SPR) was recently used in TopI-inhibition studies. However, most such immobilized small molecules or short-sequence nucleotides are used as ligands onto sensor chips, and TopI was used as the analyte that flowed through the sensor chip.

Methods

We established a sensor chip on which the TopI protein is immobilized to evaluate TopI inhibition by SPR. Camptothecin (CPT) targeting the DNA-TopI complex was used as a representative inhibitor to validate this label-free method.

Results

Purified recombinant human TopI was covalently coupled to the sensor chip for the SPR assay. The binding of anti-human (h)TopI antibodies and plasmid pUC19, respectively, to the immobilized hTopI was observed with dose-dependent increases in resonance units (RU) suggesting that the immobilized hTopI retains its DNA-binding activity. Neither CPT nor evodiamine alone in the analyte flowing through the sensor chip showed a significant increase in RU. The combination of pUC19 and TopI inhibitors as the analyte flowing through the sensor chip caused increases in RU. This confirms its reliability for binding kinetic studies of DNA-TopI binders for interaction and for primary screening of TopI inhibitors.

Conclusions

TopI immobilized on the chip retained its bioactivities of DNA binding and catalysis of intermediates of the DNA-TopI complex. This provides DNA-TopI binders for interaction and primary screening with a label-free method. In addition, this biochip can also ensure the reliability of binding kinetic studies of TopI.
RNF122: A novel ubiquitin ligase associated with calcium-modulating cyclophilin ligand
Authors: Peng Zhi , Shi Taiping , Ma Dalong
Published: June 2010

Background

RNF122 is a recently discovered RING finger protein that is associated with HEK293T cell viability and is overexpressed in anaplastic thyroid cancer cells. RNF122 owns a RING finger domain in C terminus and transmembrane domain in N terminus. However, the biological mechanism underlying RNF122 action remains unknown.

Results

In this study, we characterized RNF122 both biochemically and intracellularly in order to gain an understanding of its biological role. RNF122 was identified as a new ubiquitin ligase that can ubiquitinate itself and undergoes degradation in a RING finger-dependent manner. From a yeast two-hybrid screen, we identified calcium-modulating cyclophilin ligand (CAML) as an RNF122-interacting protein. To examine the interaction between CAML and RNF122, we performed co-immunoprecipitation and colocalization experiments using intact cells. What is more, we found that CAML is not a substrate of ubiquitin ligase RNF122, but that, instead, it stabilizes RNF122.

Conclusions

RNF122 can be characterized as a C3H2C3-type RING finger-containing E3 ubiquitin ligase localized to the ER. RNF122 promotes its own degradation in a RING finger-and proteasome-dependent manner. RNF122 interacts with CAML, and its E3 ubiquitin ligase activity was noted to be dependent on the RING finger domain.
Population-level transcriptome sequencing of nonmodel organisms Erynnis propertius and Papilio zelicaon
Authors: O'Neil Shawn T , Dzurisin Jason DK , Carmichael Rory D , Lobo Neil F , Emrich Scott J , Hellmann Jessica J
Published: June 2010

Background

Several recent studies have demonstrated the use of Roche 454 sequencing technology forde novotranscriptome analysis. Low error rates and high coverage also allow for effective SNP discovery and genetic diversity estimates. However, genetically diverse datasets, such as those sourced from natural populations, pose challenges for assembly programs and subsequent analysis. Further, estimating the effectiveness of transcript discovery using Roche 454 transcriptome data is still a difficult task.

Results

Using the Roche 454 FLX Titanium platform, we sequenced and assembled larval transcriptomes for two butterfly species: the Propertius duskywing,Erynnis propertius(Lepidoptera: Hesperiidae) and the Anise swallowtail,Papilio zelicaon(Lepidoptera: Papilionidae). The Expressed Sequence Tags (ESTs) generated represent a diverse sample drawn from multiple populations, developmental stages, and stress treatments.

Despite this diversity, > 95% of the ESTs assembled into long (> 714 bp on average) and highly covered (> 9.6× on average) contigs. To estimate the effectiveness of transcript discovery, we compared the number of bases in the hit region of unigenes (contigs and singletons) to the length of the best match silkworm (Bombyx mori) protein--this "ortholog hit ratio" gives a close estimate on the amount of the transcript discovered relative to a model lepidopteran genome. For each species, we tested two assembly programs and two parameter sets; although CAP3 is commonly used for such data, the assemblies produced by Celera Assembler with modified parameters were chosen over those produced by CAP3 based on contig and singleton counts as well as ortholog hit ratio analysis. In the final assemblies, 1,413E. propertiusand 1,940P. zelicaonunigenes had a ratio > 0.8; 2,866E. propertiusand 4,015P. zelicaonunigenes had a ratio > 0.5.

Conclusions

Ultimately, these assemblies and SNP data will be used to generate microarrays for ecoinformatics examining climate change tolerance of different natural populations. These studies will benefit from high quality assemblies with few singletons (less than 26% of bases for each assembled transcriptome are present in unassembled singleton ESTs) and effective transcript discovery (over 6,500 of our putative orthologs cover at least 50% of the corresponding model silkworm gene).
Benzo(a)pyrene induces similar gene expression changes in testis of DNA repair proficient and deficient mice
Authors: Verhofstad Nicole , Pennings Jeroen LA , van Oostrom Conny ThM , van Benthem Jan , van Schooten Frederik J , van Steeg Harry , Godschalk Roger WL
Published: June 2010

Background

Benzo [a]pyrene (B[a]P) exposure induces DNA adducts at all stages of spermatogenesis and in testis, and removal of these lesions is less efficient in nucleotide excision repair deficientXpc-/-mice than in wild type mice. In this study, we investigated by using microarray technology whether compromised DNA repair inXpc-/-mice may lead to a transcriptional reaction of the testis to cope with increased levels of B[a]P induced DNA damage.

Results

Two-Way ANOVA revealed only 4 genes differentially expressed between wild type andXpc-/-mice, and 984 genes between testes of B[a]P treated and untreated mice irrespective of the mouse genotype. However, the level in which these B[a]P regulated genes are expressed differs between Wt andXpc-/-mice (p = 0.000000141), and were predominantly involved in the regulation of cell cycle, translation, chromatin structure and spermatogenesis, indicating a general stress response. In addition, analysis of cell cycle phase dependent gene expression revealed that expression of genes involved in G1-S and G2-M phase arrest was increased after B[a]P exposure in both genotypes. A slightly higher induction of average gene expression was observed at the G2-M checkpoint inXpc-/-mice, but this did not reach statistical significance (P = 0.086). Other processes that were expected to have changed by exposure, like apoptosis and DNA repair, were not found to be modulated at the level of gene expression.

Conclusion

Gene expression in testis of untreatedXpc-/-and wild type mice were very similar, with only 4 genes differentially expressed. Exposure to benzo(a)pyrene affected the expression of genes that are involved in cell cycle regulation in both genotypes, indicating that the presence of unrepaired DNA damage in testis blocks cell proliferation to protect DNA integrity in both DNA repair proficient and deficient animals.
Comparison and calibration of transcriptome data from RNA-Seq and tiling arrays
Authors: Agarwal Ashish , Koppstein David , Rozowsky Joel , Sboner Andrea , Habegger Lukas , Hillier LaDeana W , Sasidharan Rajkumar , Reinke Valerie , Waterston Robert H , Gerstein Mark
Published: June 2010

Background

Tiling arrays have been the tool of choice for probing an organism's transcriptome without prior assumptions about the transcribed regions, but RNA-Seq is becoming a viable alternative as the costs of sequencing continue to decrease. Understanding the relative merits of these technologies will help researchers select the appropriate technology for their needs.

Results

Here, we compare these two platforms using a matched sample of poly(A)-enriched RNA isolated from the second larval stage of C. elegans. We find that the raw signals from these two technologies are reasonably well correlated but that RNA-Seq outperforms tiling arrays in several respects, notably in exon boundary detection and dynamic range of expression. By exploring the accuracy of sequencing as a function of depth of coverage, we found that about 4 million reads are required to match the sensitivity of two tiling array replicates. The effects of cross-hybridization were analyzed using a "nearest neighbor" classifier applied to array probes; we describe a method for determining potential "black list" regions whose signals are unreliable. Finally, we propose a strategy for using RNA-Seq data as a gold standard set to calibrate tiling array data. All tiling array and RNA-Seq data sets have been submitted to the modENCODE Data Coordinating Center.

Conclusions

Tiling arrays effectively detect transcript expression levels at a low cost for many species while RNA-Seq provides greater accuracy in several regards. Researchers will need to carefully select the technology appropriate to the biological investigations they are undertaking. It will also be important to reconsider a comparison such as ours as sequencing technologies continue to evolve.
TRial of an Educational intervention on patients' knowledge of Atrial fibrillation and anticoagulant therapy, INR control, and outcome of Treatment with warfarin (TREAT)
Authors: Smith Danielle E , Xuereb Christian Borg , Pattison Helen M , Lip Gregory YH , Lane Deirdre A
Published: June 2010

Background

Atrial fibrillation (AF) patients with a high risk of stroke are recommended anticoagulation with warfarin. However, the benefit of warfarin is dependent upon time spent within the target therapeutic range (TTR) of their international normalised ratio (INR) (2.0 to 3.0). AF patients possess limited knowledge of their disease and warfarin treatment and this can impact on INR control. Education can improve patients' understanding of warfarin therapy and factors which affect INR control.

Methods/Design

Randomised controlled trial of an intensive educational intervention will consist of group sessions (between 2-8 patients) containing standardised information about the risks and benefits associated with OAC therapy, lifestyle interactions and the importance of monitoring and control of their International Normalised Ratio (INR). Information will be presented within an 'expert-patient' focussed DVD, revised educational booklet and patient worksheets. 200 warfarin-naïve patients who are eligible for warfarin will be randomised to either the intervention or usual care groups. All patients must have ECG-documented AF and be eligible for warfarin (according to the NICE AF guidelines). Exclusion criteria include: aged < 18 years old, contraindication(s) to warfarin, history of warfarin USE, valvular heart disease, cognitive impairment, are unable to speak/read English and disease likely to cause death within 12 months. Primary endpoint is time spent in TTR. Secondary endpoints include measures of quality of life (AF-QoL-18), anxiety and depression (HADS), knowledge of AF and anticoagulation, beliefs about medication (BMQ) and illness representations (IPQ-R). Clinical outcomes, including bleeding, stroke and interruption to anticoagulation will be recorded. All outcome measures will be assessed at baseline and 1, 2, 6 and 12 months post-intervention.

Discussion

More data is needed on the clinical benefit of educational intervention with AF patients receiving warfarin.

Trial registration

ISRCTN93952605
The impact of CFNS-causing EFNB1 mutations on ephrin-B1 function
Authors: Makarov Roman , Steiner Bernhard , Gucev Zoran , Tasic Velibor , Wieacker Peter , Wieland Ilse
Published: June 2010

Background

Mutations of EFNB1 cause the X-linked malformation syndrome craniofrontonasal syndrome (CFNS). CFNS is characterized by an unusual phenotypic pattern of inheritance, because it affects heterozygous females more severely than hemizygous males. This sex-dependent inheritance has been explained by random X-inactivation in heterozygous females and the consequences of cellular interference of wild type and mutant EFNB1-expressing cell populations. EFNB1 encodes the transmembrane protein ephrin-B1, that forms bi-directional signalling complexes with Eph receptor tyrosine kinases expressed on complementary cells. Here, we studied the effects of patient-derived EFNB1 mutations predicted to give rise to truncated ephrin-B1 protein or to disturb Eph/ephrin-B1 reverse ephrin-B1 signalling. Five mutations are investigated in this work: nonsense mutation c.196C > T/p.R66X, frameshift mutation c.614_615delCT, splice-site mutation c.406 + 2T > C and two missense mutations p.P54L and p.T111I. Both missense mutations are located in the extracellular ephrin domain involved in Eph-ephrin-B1 recognition and higher order complex formation.

Methods

Nonsense mutation c.196C > T/p.R66X, frameshift mutation c.614_615delCT and splice-site mutation c.406+2T > C were detected in the primary patient fibroblasts by direct sequencing of the DNA and were further analysed by RT-PCR and Western blot analyses.

The impact of missense mutations p.P54L and p.T111I on cell behaviour and reverse ephrin-B1 cell signalling was analysed in a cell culture model using NIH 3T3 fibroblasts. These cells were transfected with the constructs generated by in vitro site-directed mutagenesis. Investigation of missense mutations was performed using the Western blot analysis and time-lapse microscopy.

Results and Discussion

Nonsense mutation c.196C > T/p.R66X and frameshift mutation c.614_615delCT escape nonsense-mediated RNA decay (NMD), splice-site mutation c.406+2T > C results in either retention of intron 2 or activation of a cryptic splice site in exon 2. However, c.614_615delCT and c.406+2T > C mutations were found to be not compatible with production of a soluble ephrin-B1 protein. Protein expression of the p.R66X mutation was predicted unlikely but has not been investigated.

Ectopic expression of p.P54L ephrin-B1 resists Eph-receptor mediated cell cluster formation in tissue culture and intracellular ephrin-B1 Tyr324 and Tyr329 phosphorylation. Cells expressing p.T111I protein show similar responses as wild type expressing cells, however, phosphorylation of Tyr324 and Tyr329 is reduced.

Conclusions

Pathogenic mechanisms in CFNS manifestation include impaired ephrin-B1 signalling combined with cellular interference.
Home-based voluntary HIV counselling and testing found highly acceptable and to reduce inequalities
Authors: Mutale Wilbroad , Michelo Charles , Jürgensen Marte , Fylkesnes Knut
Published: June 2010

Background

Low uptake of voluntary HIV counselling and testing (VCT) in sub-Saharan Africa is raising acceptability concerns which might be associated with ways by which it is offered. We investigated the acceptability of home-based delivery of counselling and HIV testing in urban and rural populations in Zambia where VCT has been offered mostly from local clinics.

Methods

A population-based HIV survey was conducted in selected communities in 2003 (n = 5035). All participants stating willingness to be HIV tested were offered VCT at home and all counselling was conducted in the participants' homes. In the urban area post-test counselling and giving of results were done the following day whereas in rural areas this could take 1-3 weeks.

Results

Of those who indicated willingness to be HIV tested, 76.1% (95%CI 74.9-77.2) were counselled and received the test result. Overall, there was an increase in the proportion ever HIV tested from 18% before provision of home-based VCT to 38% after. The highest increase was in rural areas; among young rural men aged 15-24 years up from 14% to 42% vs. for urban men from 17% to 37%. Test rates by educational attainment changed from being positively associated to be evenly distributed after home-based VCT.

Conclusions

A high uptake was achieved by delivering HIV counselling and testing at home. The highest uptakes were seen in rural areas, in young people and groups with low educational attainment, resulting in substantial reductions in existing inequalities in accessing VCT services.
Production of recombinant cholesterol oxidase containing covalently bound FAD in Escherichia coli
Authors: Volontè Federica , Pollegioni Loredano , Molla Gianluca , Frattini Luca , Marinelli Flavia , Piubelli Luciano
Published: June 2010

Background

Cholesterol oxidase is an alcohol dehydrogenase/oxidase flavoprotein that catalyzes the dehydrogenation of C(3)-OH of cholesterol. It has two major biotechnological applications, i.e. in the determination of serum (and food) cholesterol levels and as biocatalyst providing valuable intermediates for industrial steroid drug production. Cholesterol oxidases of type I are those containing the FAD cofactor tightly but not covalently bound to the protein moiety, whereas type II members contain covalently bound FAD. This is the first report on the over-expression in Escherichia coli of type II cholesterol oxidase from Brevibacterium sterolicum (BCO).

Results

Design of the plasmid construct encoding the mature BCO, optimization of medium composition and identification of the best cultivation/induction conditions for growing and expressing the active protein in recombinant E. coli cells, concurred to achieve a valuable improvement: BCO volumetric productivity was increased from ~500 up to ~25000 U/L and its crude extract specific activity from 0.5 up to 7.0 U/mg protein. Interestingly, under optimal expression conditions, nearly 55% of the soluble recombinant BCO is produced as covalently FAD bound form, whereas the protein containing non-covalently bound FAD is preferentially accumulated in insoluble inclusion bodies.

Conclusions

Comparison of our results with those published on non-covalent (type I) COs expressed in recombinant form (either in E. coli or Streptomyces spp.), shows that the fully active type II BCO can be produced in E. coli at valuable expression levels. The improved over-production of the FAD-bound cholesterol oxidase will support its development as a novel biotool to be exploited in biotechnological applications.
Accurate non-invasive image-based cytotoxicity assays for cultured cells
Authors: Marqués-Gallego Patricia , den Dulk Hans , Backendorf Claude , Brouwer Jaap , Reedijk Jan , Burke Julian F
Published: June 2010

Background

The CloneSelect™ Imager system is an image-based visualisation system for cell growth assessment. Traditionally cell proliferation is measured with the colorimetric MTT assay.

Results

Here we show that both the CloneSelect Imager and the MTT approach result in comparable EC50 values when assaying the cytotoxicity of cisplatin and oxaliplatin on various cell lines. However, the image-based technique was found non-invasive, considerably quicker and more accurate than the MTT assay.

Conclusions

This new image-based technique has the potential to replace the cumbersome MTT assay when fast, unbiased and high-throughput cytotoxicity assays are requested.
Extraction of single-trial cortical beta oscillatory activities in EEG signals using empirical mode decomposition
Authors: Yeh Chia-Lung , Chang Hsiang-Chih , Wu Chi-Hsun , Lee Po-Lei
Published: June 2010

Background

Brain oscillatory activities are stochastic and non-linearly dynamic, due to their non-phase-locked nature and inter-trial variability. Non-phase-locked rhythmic signals can vary from trial-to-trial dependent upon variations in a subject's performance and state, which may be linked to fluctuations in expectation, attention, arousal, and task strategy. Therefore, a method that permits the extraction of the oscillatory signal on a single-trial basis is important for the study of subtle brain dynamics, which can be used as probes to study neurophysiology in normal brain and pathophysiology in the diseased.

Methods

This paper presents an empirical mode decomposition (EMD)-based spatiotemporal approach to extract neural oscillatory activities from multi-channel electroencephalograph (EEG) data. The efficacy of this approach manifests in extracting single-trial post-movement beta activities when performing a right index-finger lifting task. In each single trial, an EEG epoch recorded at the channel of interest (CI) was first separated into a number of intrinsic mode functions (IMFs). Sensorimotor-related oscillatory activities were reconstructed from sensorimotor-related IMFs chosen by a spatial map matching process. Post-movement beta activities were acquired by band-pass filtering the sensorimotor-related oscillatory activities within a trial-specific beta band. Signal envelopes of post-movement beta activities were detected using amplitude modulation (AM) method to obtain post-movement beta event-related synchronization (PM-bERS). The maximum amplitude in the PM-bERS within the post-movement period was subtracted by the mean amplitude of the reference period to find the single-trial beta rebound (BR).

Results

The results showed single-trial BRs computed by the current method were significantly higher than those obtained from conventional average method (P < 0.01; matched-pair Wilcoxon test). The proposed method provides high signal-to-noise ratio (SNR) through an EMD-based decomposition and reconstruction process, which enables event-related oscillatory activities to be examined on a single-trial basis.

Conclusions

The EMD-based method is effective for artefact removal and extracting reliable neural features of non-phase-locked oscillatory activities in multi-channel EEG data. The high extraction rate of the proposed method enables the trial-by-trial variability of oscillatory activities can be examined, which provide a possibility for future profound study of subtle brain dynamics.
Using dynamic pupillometry as a simple screening tool to detect autonomic neuropathy in patients with diabetes: a pilot study
Authors: Ferrari Giselle L , Marques Jefferson LB , Gandhi Rajiv A , Heller Simon R , Schneider Fábio K , Tesfaye Solomon , Gamba Humberto R
Published: June 2010

Background

Autonomic neuropathy is a common and serious complication of diabetes. Early detection is essential to enable appropriate interventional therapy and management. Dynamic pupillometry has been proposed as a simpler and more sensitive tool to detect subclinical autonomic dysfunction. The aim of this study was to investigate pupil responsiveness in diabetic subjects with and without cardiovascular autonomic neuropathy (CAN) using dynamic pupillometry in two sets of experiments.

Methods

During the first experiment, one flash was administered and the pupil response was recorded for 3 s. In the second experiment, 25 flashes at 1-s interval were administered and the pupil response was recorded for 30 s. Several time and pupil-iris radius-related parameters were computed from the acquired data. A total of 24 diabetic subjects (16 without and 8 with CAN) and 16 healthy volunteers took part in the study.

Results

Our results show that diabetic subjects with and without CAN have sympathetic and parasympathetic dysfunction, evidenced by diminished amplitude reflexes and significant smaller pupil radius. It suggests that pupillary autonomic dysfunction occurs before a more generalized involvement of the autonomic nervous system, and this could be used to detect early autonomic dysfunction.

Conclusions

Dynamic pupillometry provides a simple, inexpensive, and noninvasive tool to screen high-risk diabetic patients for diabetic autonomic neuropathy.
Second-opinion stress tele-echocardiography for the Adonhers (Aged donor heart rescue by stress echo) project
Authors: Franchi Daniele , Cini Davide , Arpesella Giorgio , Gherardi Sonia , Calamai Italo , Barletta Giuseppe , Valente Serafina , Pasanisi Emilio , Sansoni Stefania , Ricci Caterina , Serra Walter , Picano Eugenio , Bombardini Tonino
Published: June 2010

Background

To resolve the current shortage of donor hearts, we established the Adonhers protocol. An upward shift of the donor age cut-off limit (from the present 55 to 65 years) is acceptable if a stress echo screening on the candidate donor heart is normal. This study aimed to verify feasibility of a "second opinion" of digitally transferred images of stress echo results to minimize technical variability in selection of aged donor hearts for heart transplant.

Methods

The informatics infrastructure was created for a core lab reading with a second opinion from the Pisa stress echo lab. To test the system, simulation standard stress echo cineloops were sent digitally from 5 peripheral labs to the central core lab.

Starting January 2009, real marginal donor stress echos were sent via internet to the central core echo lab, Pisa, for a second opinion before heart transplant.

Results

In the simulation protocol, 30 dipyridamole stress echocardiograms were sent from the five peripheral echo labs to the central core lab in Pisa. Both the echo images and reports were correctly uploaded in the web system and sent to the core echo lab; the second opinion evaluation was obtained in all cases (100% feasibility). In the transplant protocol, eight donor cases were sent to the Pisa core lab for the second opinion protocol, and six of them were transplanted in marginal recipients.

Conclusions

Second-Opinion Stress Tele-Echocardiography can effectively be performed in a network aimed to safely expand the heart donor pool for heart transplant.
Comparison of the inhibitory effects of three transcriptional variants of CDKN2A in human lung cancer cell line A549
Authors: Zhang Wei , Zhu Jing , Bai Jing , Jiang Hui , Liu Fangli , Liu An , Liu Peng , Ji Guohua , Guan Rongwei , Sun Donglin , Ji Wei , Yu Yang , Jin Yan , Meng Xiangning , Fu Songbin
Published: June 2010

Background

The tumor suppressor gene CDKN2A generates at least three different transcriptional variants, each of which is thought to encode a tumor suppressor. However, the inhibitory activities of these variants have not yet been compared in the same cells. Protein therapy is known to have several advantages over gene therapy. Thus, investigation of the exogenous protein molecule of the most effective suppressor may yield meaningful information regarding protein-based cancer therapy.

Methods

The inhibitory effects of p16INK4a, p14ARF and p12 were studied in the human lung cancer cell line A549 which lacks the CDKN2A locus. The eukaryotic expression plasmids of the three transcriptional variants were constructed and stably transfected into the cells. RNA and protein expression by the plasmids was confirmed using RT-PCR and fluorescence immunocytochemistry, respectively. Cell growth inhibition and cell-cycle redistribution after transfection were investigated based on growth curve and flow cytometry analyses. An exogenous His-tag fusion p16INK4a protein was obtained and purified by affinity chromatography. Cell growth inhibition and cell cycle arrest induced by the expression of p16INK4a protein were measured in A549 cells transduced with the exogenous protein.

Results

While all three variants suppressed cell growth, p16INK4a had the strongest effect. Marked G1-phase accumulation and S-phase inhibition were induced by p16INK4a and p14ARF but not by p12. Exogenous p16INK4a protein was successfully expressed and purified and transduction of the fusion protein into A549 cells inhibited cell growth by G1→S arrest.

Conclusions

Among the three transcript variants, p16INK4a has a greater inhibitory effect than p14ARF and p12; exogenous p16INK4a protein should be further investigated for use in cancer therapy as a protein agent.
Anti-tumor activity of N-trimethyl chitosan-encapsulated camptothecin in a mouse melanoma model
Authors: Liu Xian-ping , Zhou Sheng-tao , Li Xing-yi , Chen Xian-cheng , Zhao Xia , Qian Zhi-yong , Zhou Li-na , Li Zhi-yong , Wang Yu-mei , Zhong Qian , Yi Tao , Li Zheng-yu , He Xiang , Wei Yu-quan
Published: June 2010

Background

Camptothecin (CPT) has recently attracted increasing attention as a promising anticancer agent for a variety of tumors. But the clinical application is largely hampered by its extreme water insolubility and unpredictable side effect. It is essential to establish an efficient and safe protocol for the administration of CPT versus melanoma.

Methods

Camptothecin was encapsulated with N-trimethyl chitosan (CPT-TMC) through microprecipitation and sonication. Its inhibition effect on B16-F10 cell proliferation and induction of apoptosis was evaluated by MTT assay and flow cytometric analysis in vitro. The anti-tumor activity of CPT-TMC was evaluated in C57BL/6 mice bearing B16-F10 melanoma. Tumor volume, tumor weight and survival time were recorded. Assessment of apoptotic cells within tumor tissue was performed by TUNEL assay. Antiangiogenesis and antiproliferation effects of CPT-TMC in vivo were conducted via CD31 and PCNA immunohistochemistry, respectively.

Results

CPT-TMC efficiently inhibited B16-F10 cells proliferation and increased apoptosis in vitro. Experiment group showed significant inhibition compared with free CPT-treated group (81.3% vs. 56.9%) in the growth of B16-F10 melanoma xenografts and prolonged the survival time of the treated mice (P < 0.05). Decreased cell proliferation, increased tumor apoptosis as well as a reduction in angiogenesis were observed.

Conclusions

Our data suggest that N-trimethyl chitosan-encapsulated camptothecin is superior to free CPT by overcoming its insolubility and finally raises the potential of its application in melanoma therapy.