bluuebunny/crossref_metadata_2025 huggingface dataset:
Analysis of the Effect of the Oral Contraceptive Pill on Clinical Outcomes in Women with Ankylosing Spondylitis
Authors: Mahendira Dharini , Thavaneswaran Arane , Carty Adele , Haroon Nigil , Anton Ammepa , Passalent Laura , Alnaqbi Khalid A. , Savage Laurie , Aslanyan Elin , Inman Robert D.
Published: June 2014

Objective.

There are unexplained sex-specific changes in the clinical expression of ankylosing spondylitis (AS). We sought to examine the potential effect of exogenous estrogen in the form of oral contraceptive pills (OCP) on AS initiation and severity.

Methods.

This cross-sectional study consisted of women with AS from the membership of the Spondylitis Association of America. Measures of disease severity included use of biological agents and hip replacement surgery, while Bath AS Functional Index (BASFI) scores served as a surrogate marker of disability. Information was obtained using a patient questionnaire on patient demographics, OCP use, pregnancy history, AS duration, medication use, and hip replacement.

Results.

There were 571 women with AS who participated in our study, consisting of 448 OCP ever-users and 123 non-OCP users. The mean age of OCP users was 42.7 yrs (± 11.5) and of non-OCP users, 48.4 yrs (± 12.1). No difference was noted in the age at initial onset of back pain. However, OCP users were significantly younger at the time of diagnosis of AS (36.5 yrs vs 39.1 yrs, p = 0.02). There were no significant differences between the 2 groups in tumor necrosis factor inhibitor or opioid use, BASFI scores, pregnancy complications, or hip surgery.

Conclusion.

The use of exogenous estrogens in the form of OCP is not associated with a measurable effect on initiation or severity of AS. Biologic and social factors may contribute to earlier diagnosis of AS in OCP users. This is the largest study to date investigating the potential effect of exogenous estrogens in women with AS.
English Language Proficiency, Health Literacy, and Trust in Physician Are Associated with Shared Decision Making in Rheumatoid Arthritis
Authors: Barton Jennifer L. , Trupin Laura , Tonner Chris , Imboden John , Katz Patricia , Schillinger Dean , Yelin Edward
Published: June 2014

Objective.

Treat-to-target guidelines promote shared decision making (SDM) in rheumatoid arthritis (RA). Also, because of high cost and potential toxicity of therapies, SDM is central to patient safety. Our objective was to examine patterns of perceived communication around decision making in 2 cohorts of adults with RA.

Methods.

Data were derived from patients enrolled in 1 of 2 longitudinal, observational cohorts [University of California, San Francisco (UCSF) RA Cohort and RA Panel Cohort]. Subjects completed a telephone interview in their preferred language that included a measure of patient-provider communication, including items about decision making. Measures of trust in physician, education, and language proficiency were also asked. Logistic regression was performed to identify correlates of suboptimal SDM communication. Analyses were performed on each sample separately.

Results.

Of 509 patients across 2 cohorts, 30% and 32% reported suboptimal SDM communication. Low trust in physician was independently associated with suboptimal SDM communication in both cohorts. Older age and limited English proficiency were independently associated with suboptimal SDM in the UCSF RA Cohort, as was limited health literacy in the RA Panel Cohort.

Conclusion.

This study of over 500 adults with RA from 2 demographically distinct cohorts found that nearly one-third of subjects report suboptimal SDM communication with their clinicians, regardless of cohort. Lower trust in physician was independently associated with suboptimal SDM communication in both cohorts, as was limited English language proficiency and older age in the UCSF RA Cohort and limited health literacy in the RA Panel Cohort. These findings underscore the need to examine the influence of SDM on health outcomes in RA.
It's not all about the music: Online fan communities and collecting Hard Rock Café pins
Authors: Geraghty Lincoln
Published: June 2014

Previous studies of music fan culture have largely centered on the diverse range of subcultures devoted to particular genres, groups, and stars. Where studies have moved beyond the actual music and examined the fashion, concerts, and collecting ephemera such as vinyl records and posters, they have tended to remain closely allied to notions of subcultural distinction, emphasizing hierarchies of taste. This paper shifts the focus in music fan studies beyond the appreciation of the music and discusses the popular fan practice of collecting souvenir pins produced and sold by the Hard Rock Café (HRC) within a framework of fan tourism. Traveling to and collecting unique pins from locations across the globe creates a fan dialogue that centers on tourism and the collecting practices associated with souvenir consumption. Collectors engage in practices such as blogging, travel writing, and administration that become important indicators of their particular expression of fandom: pin collecting. Membership requires both time and money; recording visits around the world and collecting unique pins from every café builds fans' cultural capital. This indicates an internationalization of popular fandom, with the Internet acting as a connective virtual space between local and national, personal and public physical space. The study of HRC pin collecting and its fan community suggests that HRC enthusiasts are not so because they enjoy rock music or follow any particular artist but due to the physical ephemera that they collect and the places and spaces they visit.
butcher, the baker, the lightsaber maker
Authors: Phillips Forrest
Published: June 2014

Lightsaber artisanship is an entrepreneurial fandom that has thus far avoided encountering legal trouble because of their deferential attitude toward the IP holder.
heterogeneity of maid cafés: Exploring object-oriented fandom in Japan
Authors: Sharp Luke
Published: June 2014

Maid cafés may be positioned as objects of (often oversimplified) fan communities in Japan; their heterogeneous qualities may be explored by presenting chronotopes with which they may be reconsidered. The problematics of viewing maid cafés in a homogeneous or reductive way (via iyashi-kei, "relaxation purposes," and moe-kei, "entertainment purposes") are considered, and the results of empirical research conducted at establishments across Honshū in Japan are presented. The vast differences in the services and interactivities provided in maid café settings are at odds with the homogenized fashion in which these venues have been treated in the popular press, in local promotional publications, and often in the academic literature. By presenting chronotopes and case studies with which maid cafés may be reconsidered, what it means to discuss maid cafés as a holistic phenomenon is challenged, and a heuristic for examining how spaces are constructed inside such establishments is provided.
invisible teenager: Comic book materiality and the amateur films of Don Glut
Authors: Yockey Matt
Published: June 2014

Don Glut, between the ages of 9 and 25, made 41 short amateur films inspired by horror, science fiction, and superhero movies, serials, and comic books. The tactile qualities of comic books as affect-generating objects are instrumental to how Glut confirmed his identity during a time (adolescence) in which that identity is particularly unstable. Glut used the popular figure of the teen rebel and his role as a filmmaker in order to negotiate with hegemonic restrictions on his objects of affection, especially comic books.
Interview with Mark Racop
Authors: Yockey Matt
Published: June 2014

An interview with replica Batmobile maker Mark Racop.
Interview with Kandy Fong
Authors: Coppa Francesca
Published: June 2014

Video interview and transcription of subject Kandy Fong, an early Star Trek vidder. Part of the OTW Oral History Project (http://transformativeworks.org/projects/oral-history).
"Send in the clones: A cultural study of the tribute band," by Georgina Gregory
Authors: Hong Sun-ha
Published: June 2014

Review of Georgina Gregory.  Send in the clones: A cultural study of the tribute band. Sheffield: Equinox, 2012. Paperback $29.95 (184p) ISBN 978-1845532451.
Beyond souvenirs: Making fannish items by hand
Authors: Bode Dana Sterling
Published: June 2014

Fan crafts such as knitting, plushies, painted objects, and food are a fulfilling and creative endeavor for the artists. Five fan crafters were interviewed via e-mail to discuss how and why they create.
"Cult collectors: Nostalgia, fandom and collecting popular culture," by Lincoln Geraghty
Authors: Duffy Michael S.
Published: June 2014

Review of Lincoln Geraghty.  Cult collectors: Nostalgia, fandom and collecting popular culture. New York: Routledge, 2014. Paperback, $39.95 (224p) ISBN 978-0-415-61766-6..
Estudio de la influencia de la automatización en el proyecto arquitectónico
Authors: Millán-Anglés S. , García-Santos A. , Jiménez-Leube F. J. , Higuera-Rincón O.
Published: June 2014

La domótica integrada puede terminar jugando un papel fundamental en la arquitectura del futuro. Sin embargo, hasta hoy la automatización doméstica no es un factor tenido en cuenta desde el proyecto arquitectónico y este hecho hace que esta disciplina no se generalice para replantear el futuro de estas nuevas construcciones. Por ello, se plantea la necesidad de un estudio profundo de la influencia sobre los aspectos constructivos y proyectuales que existen en la domotización y automatización de viviendas. Para desarrollar esta investigación se han aislado los posibles condicionantes de la automatización, condicionantes que deberían tenerse en cuenta desde la fase proyecto para fomentar la implantación de las nuevas tecnologías en los edificios y así garantizar la viabilidad de estas nuevas instalaciones. El presente artículo analiza, por tanto, las principales consideraciones de carácter constructivo y proyectual que se están detectando en el estudio de la influencia de la domótica sobre la forma de concebir un proyecto arquitectónico.
Inverted Classroom by Topic - A Study in Mathematics for Electrical Engineering Students
Authors: Braun Isabel , Ritter Stefan , Vasko Mikko
Published: June 2014

The inverted classroom is a teaching model, where the students prepare for classroom by watching video lectures. The classroom time is then dedicated to individual practice. We evaluated a mathematics course for electrical engineering students throughout three semesters, where 20% of the topics were taught using the inverted classroom model. The aim was to find out whether the model can help to better address groups with large differences in prior knowledge in mathematics. We report mainly positive feedback from the students, although the opinions vary greatly between the groups. The students appreciate the increased amount of practice in the classroom as well as the possibility to learn at their own pace. Exam performance remained constant in the topics taught using the inverted classroom compared to previous semesters. The exam performance of weaker students also remained constant.
On-professional Competences in Engineering Education for XL-Classes
Authors: Schröder Stefan , Janssen Daniela , Leisten Ingo , Vossen René , Isenhardt Ingrid
Published: June 2014

Far reaching changes in university higher education have taken place in the last ten years. Different factors, e.g. necessity of on-professional competences in engineering education, rising or vast student numbers and new technical possibilities, have influenced the academic teaching and learning process. Therefore interdependence between requirements and didactical-educational possibilities is given. Because of changed circumstances an adaption of teaching methods and concepts is required. At the same time Bologna arrogates students to be placed in the centre of the teaching and learning process and claims on-professional competences for today's students. Especially for XL-Classes this is a specific challenge. One of the questions ensuing is how to increase learning success by the use of specific didactical methods? With a research approach connecting different proven didactical concepts and considering the previously shown conditions, the concept of the lecture ?communication and organizational development (KOE) at RWTH Aachen University has been redesigned. This lecture, organized by the Institute Cluster IMA/ZLW & IfU at RWTH Aachen University, is mainly frequented by up to nearly 1.300 students of the faculty of mechanical engineering and inherent part of the bachelor-curriculum. The following practical example prospects the multi-angulation of didactical concepts and shows up innovative educational teaching.
Personalised Learning Object System Based on Self-Regulated Learning Theories
Authors: Alharbi Ali , Henskens Frans , Hannaford Michael
Published: June 2014

Self-regulated learning has become an important construct in education research in the last few years. Selfregulated learning in its simple form is the learner’s ability to monitor and control the learning process. There is increasing research in the literature on how to support students become more self-regulated learners. However, the advancement in the information technology has led to paradigm changes in the design and development of educational content. The concept of learning object instructional technology has emerged as a result of this shift in educational technology paradigms. This paper presents the results of a study that investigated the potential educational effectiveness of a pedagogical framework based on the self-regulated learning theories to support the design of learning object systems to help computer science students. A prototype learning object system was developed based on the contemporary research on self-regulated learning. The system was educationally evaluated in a quasi-experimental study over two semesters in a core programming languages concepts course. The evaluation revealed that a learning object system that takes into consideration contemporary research on self-regulated learning can be an effective learning environment to support computer science education.
Engineering Students as Science Teachers: A Case Study on Students' Motivation
Authors: Gero Aharon
Published: June 2014

The program "Educational Clinic" was recently developed and implemented at the Technion – Israel Institute of Technology. This one year program is designed to train engineering students as teaching assistants in high schools in order to help high school pupils with mathematics and science. The study described in this paper tracked changes in students' motivation to participate in the program throughout the year. Data was collected by questionnaires and interviews. The findings reveal that alongside a fixed high level of extrinsic motivational factors, which reflect student satisfaction of improving their teaching skills, a considerable increase was found in the level of intrinsic motivational factors, which express the students' interest in the program.
Learning Tools and Applications for Cognitive Improvement
Authors: Drigas Athanasios , Karyotaki Maria
Published: June 2014

Learning technologies are an indispensable tool for students’ cognitive improvement and assessment. ICTs in coordination with a concrete pedagogical framework may provide students and teachers flexible, engaging, cost-effective and above all, personalized learning experiences, which focus on the adoption of the 21st century cognitive skills into the actual learning process. Such higher order thinking skills (HOTS) entail critical thinking, problem solving, independent inquiry, creativity, communication, collaboration, digital literacy. Therefore, technologically-supported educational environments aim at self-regulated and inquisitive, constructivist, knowledge building rather than knowledge accumulation.
Image Fusion and Evaluation of Geological Disaster Based on Remote Sensing
Authors: Wang Juan , Wang Huajun , Li Yinghao , Chen Hairui
Published: June 2014

The investigation of geological disaster in our article locates in southern Australia, which is characterized by wide range, high relief, inaccessibility and other unfavorable factors. Multi-spectral ETM+ and SPOT 5 pan images were selected as the remote sensing data source, and Brovey transform (BT), intensity-hue-saturation (IHS), principal component analysis (PCA), high-pass filtering (HPF) and modified Gram-Schmidt (MGS) methods were used for image fusion. A comparison has been conducted between the resultant fusion images to assess the image quality both in subjective and objective evaluation. The results show that, the MGS method is the optimal image fusion method for geological disaster interpretation, and can provide abundant textural and spectral information in interpreting such geological disasters as landslide, rock fall and debris flow.
Interactive White Boards in Preschool and Primary Education
Authors: Drigas Athanasios , Papanastasiou George
Published: June 2014

Technologies within the domain of interactive, remote and on line science which are extensively adopted in education’s everyday life are interactive whiteboards and related applications. Interactive whiteboards indicate positive effects on students’ learning and instructors' teaching, promoting whole class teaching. In this review-paper we cope with the studies that explore the integration of IWBs in preschool and primary education in the last decade (2004-2013). Research has shown that interactive whiteboards are able to keep students involved and foster their attention in every aspect of the curriculum, much easier than without it. This technology has proven its value added to students and teachers in different facets of teaching and learning, in literacy, in mathematics, in science, in physics, its impact on English Language Learners, on relations and learning, on the use of IWBs by kindergartners, professional development of teachers, pedagogical orchestration, classroom dialogue and pedagogic practice.
AN EFFICIENT RIPPLET-BASED SHRINKAGE TECHNIQUE FOR MR IMAGE RESTORATION
Authors: Begum A. Sumaiya , Poornachandra S.
Published: June 2014

In this paper a new ripplet-based shrinkage technique is used to suppress noise from Magnetic Resonance Imaging (MRI). The propitious properties of ripplet transform such as anisotropy, high directionality, good localization, and high-energy compaction make the proposed method efficient and feature preserving when compared to other transforms. Ripplet transform provides efficient representation of edges in images with a higher potential for image processing applications such as image restoration, compression, and de-noising. The proposed method implies a new nonlinear ripplet-based shrinkage technique to extract the spatial and frequency information from MRI corrupted by noise. The choice of this new shrinkage technique is due to its simplicity, versatility, and its efficiency in removing noise from homogenous regions and those regions with singularities, when compared to the existing filtering techniques. Experiments were conducted on several diffusion weighed images and anatomical images. The results show that the proposed de-noising technique shows competitive performance compared to the current state-of-art methods. Qualitative validation was performed based on several quality metrics and profound improvement over existing methods was obtained. Higher values of Peak Signal to Noise Ratio (PSNR), Correlation Coefficient (CC), mean structural similarity index (MSSIM), and lower values of Root Mean Square Error (RMSE) and computational time were obtained for the proposed ripplet-based shrinkage technique when compared to the existing ones.
Drag Coefficients of Stock and Stockless Anchors
Authors: Woo Jinho , Na Won-Bae
Published: June 2014

AbstractIn recent years, collisions between anchors and submarine power cables have attracted increasing research interest because of the rapid growth of offshore wind farms. Since the drag coefficients have not been calculated, conservative values have been assumed and used
to simulate collisions. This conservative estimation is likely to result in unnecessarily heavy designs for the protective structures. Therefore, reliable calculations of the drag coefficients are required to reduce the design, manufacture, and installation costs of the protective structures.
Here, we describe calculations of the drag coefficients of 2,000-kg stock and stockless anchors using numerical flow analyses. We show that the drag coefficients of the anchors converge to 1.0 and 0.8, respectively, regardless of the initial velocity and the Reynolds number.
Minimal Time Route for Wind-Assisted Ships
Authors: Yuankui Li , Yingjun Zhang , Feixiang Zhu
Published: June 2014

AbstractAs a result of a global call for energy-saving and emission-reduction strategies as well as an urgent need to reduce the shipping cost of transoceanic crossings, this paper proposes a route that minimizes the time for such crossings and provides technical support
to efficiently utilize wind power based on existing research for wind-assisted ships. To begin, the ocean winds around the ship route were analyzed, and the different influences on traditional ships and wind-assisted ships were listed for various wind speeds and directions. The number of waypoints
of a route was subsequently calculated, and a model of the optimal ship route was then built based on the fixed power output of the main marine engine. A solution algorithm based on simulated annealing was then presented to determine the optimal wind-assisted ship routes by minimizing the
travel time. Finally, a 76,000-DWT wind-assisted cargo ship was designated as the experimental ship, and the optimization model and its algorithm were simulated to generate an optimized wind-assisted route. The simulation indicated that the speed of a ship equipped with wind propulsion increases,
which significantly reduces the travel time and fuel costs over the optimized route, despite the increased distance of this route. Thus, the route optimization algorithm designed in this study can be applied to optimize the routes for wind-assisted ships and theoretically guide further studies
of wind-assisted projects.
Path Replanning Method for an AUV in Natural Hydrothermal Vent Fields: Toward 3D Imaging of a Hydrothermal Chimney
Authors: Sato Yoshiki , Maki Toshihiro , Kume Ayaka , Matsuda Takumi , Sakamaki Takashi , Ura Tamaki
Published: June 2014

AbstractAutonomous underwater vehicles (AUVs) can operate without the need for human control or tether cables as long as there is sufficient energy. AUVs have recently been used for seafloor imaging. Visual observation by AUVs provides high-resolution color information of
the seafloor. However, conventional observation techniques that follow a prespecified path offer limited coverage because it is impossible for operators to build a suitable path in unknown rough terrain. A flawed prespecified path will produce incomplete observation. If unobserved areas are
found during postprocessing, another dive is necessary, which increases the total cost. To overcome this problem, the authors have proposed a path replanning method to realize high-coverage observation in one dive. With this method, the AUV evaluates unobserved areas after the first prespecified
observation; if unobserved areas are found, the AUV recreates an appropriate path to cover what was missed. The validity of the proposed method was previously evaluated using an artificial target in a tank and in shallow seas at a depth of approximately 35 m. In this study, the feasibility
of the method was validated in a more challenging setting: experimental data were taken from a hydrothermal vent field in Kagoshima Bay, Japan.
Review of Technological Advancements and HSE-Based Safety Model for Deep-Water Human Occupied Vehicles
Authors: Vedachalam Narayanaswamy , Ramadass Gidugu Ananada , Atmanand Malayath Aravindakshan
Published: June 2014

AbstractThis paper reviews the latest advancements in subsea technologies associated with the safety of deep-water human occupied vehicles. Human occupied submersible operations are required for deep-water activities, such as high-resolution bathymetry, biological and geological
surveys, search activities, salvage operations, and engineering support for underwater operations. As this involves direct human presence, the system has to be extremely safe and reliable. Based on applicable IEC 61508 Standards for health, safety, and environment (HSE), the safety integrity
level requirements for the submersible safety systems are estimated. Safety analyses are done on 10 critical submersible safety systems with the assumption that the submersible is utilized for 10 deep-water missions per year. The results of the analyses are compared with the estimated target
HSE requirements, and it is found that, with the present technological maturity and safety-centered design, it is possible to meet the required safety integrity levels. By proper maintenance, it is possible to keep the mean time between failures to more than 9 years. The results presented
shall serve as a model for designers to arrive at the required trade-off between the capital expenditure, operating expenditure, and required safety levels.
Evaluation of a Low-Cost and Accurate Ocean Temperature Logger on Subsurface Mooring Systems
Authors: Tian Chuan , Deng Zhiqun Daniel , Lu Jun , Xu Xiaoyang , Zhao Wei , Xu Ming
Published: June 2014

AbstractMonitoring seawater temperature is important to understanding evolving ocean processes. To monitor internal waves or ocean mixing, a large number of temperature loggers are typically mounted on subsurface mooring systems to obtain high-resolution temperature data
at different water depths. In this study, we redesigned and evaluated a compact, low-cost, self-contained, high-resolution and high-accuracy ocean temperature logger, the TC-1121. The newly designed TC-1121 logger is smaller and more robust, and its sampling interval can be automatically changed
by indicated events. The logger’s initial accuracy is ±0.002°C, and its effective resolution is about 0.0001°C. The drift error of all six TC-1121 temperature loggers during a 450-day experiment was less than ±0.002°C. They are being widely used in many mooring
systems to study internal wave and ocean mixing. The logger’s fundamental design, noise analysis, calibration, and drift test are discussed. A long-term sea trial in the South China Sea was completed successfully and demonstrated the effectiveness of the logger.
Targeting of TLRs Inhibits CD4+ Regulatory T Cell Function and Activates Lymphocytes in Human Peripheral Blood Mononuclear Cells
Authors: Voo Kui Shin , Bover Laura , Harline Megan Lundell , Weng Jinsheng , Sugimoto Naoshi , Liu Yong-Jun
Published: June 2014

Accumulating evidence suggests elements within tumors induce exhaustion of effector T cells and infiltration of immunosuppressive regulatory T cells (Tregs), thus preventing the development of durable antitumor immunity. Therefore, the discovery of agents that simultaneously block Treg suppressive function and reinvigorate effector function of lymphocytes is key to the development of effective cancer immunotherapy. Previous studies have shown that TLR ligands (TLRLs) could modulate the function of these T cell targets; however, those studies relied on cell-free or accessory cell-based assay systems that do not accurately reflect in vivo responses. In contrast, we used a human PBMC-based proliferation assay system to simultaneously monitor the effect of TLRLs on T cells (CD4+, CD8+, Tregs), B cells, and NK cells, which gave different and even conflicting results. We found that the TLR7/8L:CL097 could simultaneously activate CD8+ T cells, B cells, and NK cells plus block Treg suppression of T cells and B cells. The TLRLs TLR1/2L:Pam3CSK4, TLR5L:flagellin, TLR4L:LPS, and TLR8/7L:CL075 also blocked Treg suppression of CD4+ or CD8+ T cell proliferation, but not B cell proliferation. Besides CL097, TLR2L:PGN, CL075, and TLR9L:CpG-A, CpG-B, and CpG-C) were strong activators of NK cells. Importantly, we found that Pam3CSK4 could: 1) activate CD4+ T cell proliferation, 2) inhibit the expansion of IL-10+ naturally occurring FOXP3+ Tregs and induction of IL-10+ CD4+ Tregs (IL-10–producing type 1 Treg), and 3) block naturally occurring FOXP3+ Tregs suppressive function. Our results suggest these agents could serve as adjuvants to enhance the efficacy of current immunotherapeutic strategies in cancer patients.
CD40 Ligand Preferentially Modulates Immune Response and Enhances Protection against Influenza Virus
Authors: Hashem Anwar M. , Gravel Caroline , Chen Ze , Yi Yinglei , Tocchi Monika , Jaentschke Bozena , Fan Xingliang , Li Changgui , Rosu-Myles Michael , Pereboev Alexander , He Runtao , Wang Junzhi , Li Xuguang
Published: June 2014

CD40L, a key regulator of the immune system, was studied as both a targeting ligand and a molecular adjuvant in nucleoprotein (NP)-based host defense against influenza in mouse models with different genetic backgrounds. Adenoviral vectors secreting NP-CD40L fusion protein (denoted as rAd-SNP40L) afforded full protection of immunocompetent and immunocompromised mice (CD40L−/− and CD4−/−) against lethal influenza infection. Mechanistically, rAd-SNP40L preferentially induced early and persistent B cell germinal center formation, and accelerated Ig isotype-switching and Th1-skewed, NP-specific Ab response. Moreover, it drastically augmented primary and memory NP-specific CTL activity and polyfunctional CD8+ T cells. The markedly enhanced nonneutralizing Abs and CTLs significantly reduced viral burdens in the lungs of mice upon lethal virus challenge. Data generated from CD40L−/− and CD4−/− mice revealed that the protection was indeed CD40L mediated but CD4+ T cell independent, demonstrating the viability of the fusion Ags in protecting immunodeficient hosts. Notably, a single dose of rAd-SNP40L completely protected mice from lethal viral challenge 4 mo after immunization, representing the first report, to our knowledge, on NP in conjunction with a molecular adjuvant inducing a robust and long-lasting memory immune response against influenza. This platform is characterized by an increased in vivo load of CD40-targeted Ag upon the secretion of the fusion protein from adenovirus-infected cells and may represent a promising strategy to enhance the breadth, durability, and potency of Ag-specific immune responses.
DOK3 Is Required for IFN-β Production by Enabling TRAF3/TBK1 Complex Formation and IRF3 Activation
Authors: Kim Susana Soo-Yeon , Lee Koon-Guan , Chin Ching-Siang , Ng Say-Kong , Pereira Natasha Ann , Xu Shengli , Lam Kong-Peng
Published: June 2014

The downstream of kinase (DOK) family of adaptors is generally involved in the negative regulation of signaling pathways. DOK1, 2, and 3 were shown to attenuate TLR4 signaling by inhibiting Ras-ERK activation. In this study, we elucidated a novel role for DOK3 in IFN-β production. Macrophages lacking DOK3 were impaired in IFN-β synthesis upon influenza virus infection or polyinosinic-polyribocytidylic acid stimulation. In the absence of DOK3, the transcription factor IFN regulatory factor 3 was not phosphorylated and could not translocate to the nucleus to activate ifn-β gene expression. Interestingly, polyinosinic-polyribocytidylic acid–induced formation of the upstream TNFR-associated factor (TRAF) 3/TANK-binding kinase (TBK) 1 complex was compromised in dok3−/− macrophages. DOK3 was shown to bind TBK1 and was required for its activation. Furthermore, we demonstrated that overexpression of DOK3 and TBK1 could significantly enhance ifn-β promoter activity. DOK3 was also shown to bind TRAF3, and the binding of TRAF3 and TBK1 to DOK3 required the tyrosine-rich C-terminal domain of DOK3. We further revealed that DOK3 was phosphorylated by Bruton’s tyrosine kinase. Hence, DOK3 plays a critical and positive role in TLR3 signaling by enabling TRAF3/TBK1 complex formation and facilitating TBK1 and IFN regulatory factor 3 activation and the induction of IFN-β production.
Real-Time Detection of CTL Function Reveals Distinct Patterns of Caspase Activation Mediated by Fas versus Granzyme B
Authors: Li Jinzhu , Figueira Sarah K. , Vrazo Alexandra C. A. , Binkowski Brock F. , Butler Braeden L. , Tabata Yasuhiro , Filipovich Alexandra , Jordan Michael B. , Risma Kimberly A.
Published: June 2014

Activation of caspase-mediated apoptosis is reported to be a hallmark of both granzyme B– and Fas-mediated pathways of killing by CTLs; however, the kinetics of caspase activation remain undefined owing to an inability to monitor target cell–specific apoptosis in real time. We have overcome this limitation by developing a novel biosensor assay that detects continuous, protease-specific activity in target cells. Biosensors were engineered from a circularly permuted luciferase, linked internally by either caspase 3/7 or granzyme B/caspase 8 cleavage sites, thus allowing activation upon proteolytic cleavage by the respective proteases. Coincubation of murine CTLs with target cells expressing either type of biosensor led to a robust luminescent signal within minutes of cell contact. The signal was modulated by the strength of TCR signaling, the ratio of CTL/target cells, and the type of biosensor used. Additionally, the luciferase signal at 30 min correlated with target cell death, as measured by a 51Cr-release assay. The rate of caspase 3/7 biosensor activation was unexpectedly rapid following granzyme B– compared with Fas-mediated signal induction in murine CTLs; the latter appeared gradually after a 90-min delay in perforin- or granzyme B–deficient CTLs. Remarkably, the Fas-dependent, caspase 3/7 biosensor signal induced by perforin-deficient human CTLs was also detectable after a 90-min delay when measured by redirected killing. Thus, we have used a novel, real-time assay to demonstrate the distinct pattern of caspase activation induced by granzyme B versus Fas in human and murine CTLs.
Ficolin-2 Inhibits Hepatitis C Virus Infection, whereas Apolipoprotein E3 Mediates Viral Immune Escape
Authors: Zhao Yinglan , Ren Yushan , Zhang Xuping , Zhao Ping , Tao Wanyin , Zhong Jin , Li Qiao , Zhang Xiao-Lian
Published: June 2014

Human ficolin-2 (L-ficolin/p35) is a lectin-complement pathway activator that is present in normal human plasma and is associated with infectious diseases; however, little is known regarding the roles and mechanisms of ficolin-2 during chronic hepatitis C virus (HCV) infection. In this study, we found that ficolin-2 inhibits the entry of HCV at an early stage of viral infection, regardless of the viral genotype. Ficolin-2 neutralized and inhibited the initial attachment and infection of HCV by binding to the HCV envelope surface glycoproteins E1 and E2, blocking HCV attachment to low-density lipoprotein receptor (LDLR) and scavenger receptor B1, and weakly interfering with CD81 receptor attachment. However, no interference with claudin-1 and occludin receptor attachment was observed. The C-terminal fibrinogen domain (201–313 aa) of ficolin-2 was identified as the critical binding region for the HCV-E1–E2 N-glycans, playing a critical role in the anti-HCV activity. More importantly, we found that apolipoprotein E (ApoE)3, which is enriched in the low-density fractions of HCV RNA–containing particles, promotes HCV infection and inhibits ficolin-2–mediated antiviral activity. ApoE3, but not ApoE2 and ApoE4, blocked the interaction between ficolin-2 and HCV-E2. Our data suggest that the HCV entry inhibitor ficolin-2 is a novel and promising antiviral innate immune molecule, whereas ApoE3 blocks the effect of ficolin-2 and mediates an immune escape mechanism during chronic HCV infection. HCV may be neutralized using compounds directed against the lipoprotein moiety of the viral particle, and ApoE3 may be a new target to combat HCV infection.
Acute and Chronic B Cell Depletion Disrupts CD4+ and CD8+ T Cell Homeostasis and Expansion during Acute Viral Infection in Mice
Authors: Lykken Jacquelyn M. , DiLillo David J. , Weimer Eric T. , Roser-Page Susanne , Heise Mark T. , Grayson Jason M. , Weitzmann M. Neale , Tedder Thomas F.
Published: June 2014

B cells provide humoral protection against pathogens and promote cellular immunity through diverse nonclassical effector functions. To assess B cell function in promoting T cell homeostasis, mature B cells were either acutely or chronically depleted in mice using CD20 mAb. Acute B cell depletion in either 2- or 4-mo-old mice significantly reduced spleen and lymph node CD4+ and CD8+ T cell numbers, including naive, activated, and Foxp3+CD25+CD4+ regulatory T cell subsets. The numbers of IFN-γ– and TNF-α–producing T cells were also significantly reduced. Chronic B cell depletion for 6 mo in aged naive mice resulted in a 40–70% reduction in activated CD4+ and CD8+ T cell numbers and 20–50% reductions in IFN-γ–producing T cells. Therefore, B cells were necessary for maintaining naive CD4+ and CD8+ T cell homeostasis for subsequent optimal T cell expansion in young and old mice. To determine the significance of this finding, a week of B cell depletion in 4-mo-old mice was followed by acute viral infection with lymphocytic choriomeningitis virus Armstrong. Despite their expansion, activated and cytokine-producing CD4+ and CD8+ T cell numbers were still significantly reduced 1 wk later. Moreover, viral peptide-specific CD4+ and CD8+ T cell numbers and effector cell development were significantly reduced in mice lacking B cells, whereas lymphocytic choriomeningitis virus titers were dramatically increased. Thus, T cell function is maintained in B cell–depleted mice, but B cells are required for optimal CD4+ and CD8+ T cell homeostasis, activation, and effector development in vivo, particularly during responses to acute viral infection.
High Proportion of CD5+ B Cells in Infants Predicts Development of Allergic Disease
Authors: Lundell Anna-Carin , Johansen Susanne , Adlerberth Ingegerd , Wold Agnes E. , Hesselmar Bill , Rudin Anna
Published: June 2014

Delayed maturation of the immune system has been proposed to be a risk factor for development of allergy, but B cell maturation in relation to allergic disease has not been examined. B cells lose CD5 and acquire CD27 during maturation from immature via mature/naive to Ig-secreting cells and memory cells. We sought to investigate B cell maturation in relation to development of allergic disease and sensitization in the FARMFLORA birth cohort including 65 Swedish children. Total B cell numbers, proportions of CD5+ and CD27+ B cells, and levels of IgM, IgG, IgA, and IgE were measured in blood on repeated occasions from birth to 36 mo of age, and related to allergic disease and sensitization at 18 and 36 mo of age with multivariate discriminant analysis. We also compared the expression of CD24 and CD38 within CD5+ and CD5neg B cells in children and in adults. We found that infants with a high proportion of CD5+ B cells at birth and at 1 mo of age had an increased risk for having allergic disease at 18 and 36 mo of life. Further, the proportions of CD5+ B cells at 1 mo of age were inversely correlated with total IgG levels at 18 and 36 mo of age. The majority of the CD5+ B cells were of a CD24hi/+CD38hi/+ immature/naive phenotype at birth (97%), 7 y of age (95%), and in adults (86%). These results suggest that development of allergic disease is preceded by an immaturity in neonatal B cell phenotype.
B Cells Regulate CD4+ T Cell Responses to Papain following B Cell Receptor–Independent Papain Uptake
Authors: Dwyer Daniel F. , Woodruff Matthew C. , Carroll Michael C. , Austen K. Frank , Gurish Michael F.
Published: June 2014

Papain, a cysteine protease allergen with inherent adjuvant activity, induces potent IL-4 expression by T cells in the popliteal lymph nodes of mice following footpad immunization. In this study, we identify a novel, non-BCR–mediated capacity for B cells to rapidly bind and internalize papain. B cells subsequently regulate the adaptive immune response by enhancing ICOS expression on CD4+ T cells and amplifying Th2 and follicular helper T cell induction. Ab blockade of ICOS ligand, expressed by popliteal lymph node B cells, but not dendritic cells, at the peak of the response inhibits IL-4 responses in wild-type mice but not B cell–deficient mice. Thus, B cells play a critical role in amplifying adjuvant-dependent Th2 polarization following noncanonical acquisition and internalization of the cysteine protease papain.
Investigation of Soluble and Transmembrane CTLA-4 Isoforms in Serum and Microvesicles
Authors: Esposito Laura , Hunter Kara M. D. , Clark Jan , Rainbow Daniel B. , Stevens Helen , Denesha Jennifer , Duley Simon , Dawson Sarah , Coleman Gillian , Nutland Sarah , Bell Gwynneth L. , Moran Carla , Pekalski Marcin , Todd John A. , Wicker Linda S.
Published: June 2014

Expression of the CTLA-4 gene is absolutely required for immune homeostasis, but aspects of its molecular nature remain undefined. In particular, the characterization of the soluble CTLA-4 (sCTLA-4) protein isoform generated by an alternatively spliced mRNA of CTLA4 lacking transmembrane-encoding exon 3 has been hindered by the difficulty in distinguishing it from the transmembrane isoform of CTLA-4, Tm-CTLA-4. In the current study, sCTLA-4 has been analyzed using novel mAbs and polyclonal Abs specific for its unique C-terminal amino acid sequence. We demonstrate that the sCTLA-4 protein is secreted at low levels following the activation of primary human CD4+ T cells and is increased only rarely in the serum of autoimmune patients. Unexpectedly, during our studies aimed to define the kinetics of sCTLA-4 produced by activated human CD4+ T cells, we discovered that Tm-CTLA-4 is associated with microvesicles produced by the activated cells. The functional roles of sCTLA-4 and microvesicle-associated Tm-CTLA-4 warrant further investigation, especially as they relate to the multiple mechanisms of action described for the more commonly studied cell-associated Tm-CTLA-4.
Opposing Roles of Prostaglandin D2 Receptors in Ulcerative Colitis
Authors: Sturm Eva M. , Radnai Balazs , Jandl Katharina , Stančić Angela , Parzmair Gerald P. , Högenauer Christoph , Kump Patrizia , Wenzl Heimo , Petritsch Wolfgang , Pieber Thomas R. , Schuligoi Rufina , Marsche Gunther , Ferreirós Nerea , Heinemann Akos , Schicho Rudolf
Published: June 2014

Proresolution functions were reported for PGD2 in colitis, but the role of its two receptors, D-type prostanoid (DP) and, in particular, chemoattractant receptor homologous molecule expressed on Th2 cells (CRTH2), is less well defined. We investigated DP and CRTH2 expression and function during human and murine ulcerative colitis (UC). Expression of receptors was measured by flow cytometry on peripheral blood leukocytes and by immunohistochemistry and immunoblotting in colon biopsies of patients with active UC and healthy individuals. Receptor involvement in UC was evaluated in a mouse model of dextran sulfate sodium colitis. DP and CRTH2 expression changed in leukocytes of patients with active UC in a differential manner. In UC patients, DP showed higher expression in neutrophils but lower in monocytes as compared with control subjects. In contrast, CRTH2 was decreased in eosinophils, NK, and CD3+ T cells but not in monocytes and CD3+/CD4+ T cells. The decrease of CRTH2 on blood eosinophils clearly correlated with disease activity. DP correlated positively with disease activity in eosinophils but inversely in neutrophils. CRTH2 internalized upon treatment with PGD2 and 11-dehydro TXB2 in eosinophils of controls. Biopsies of UC patients revealed an increase of CRTH2-positive cells in the colonic mucosa and high CRTH2 protein content. The CRTH2 antagonist CAY10595 improved, whereas the DP antagonist MK0524 worsened inflammation in murine colitis. DP and CRTH2 play differential roles in UC. Although expression of CRTH2 on blood leukocytes is downregulated in UC, CRTH2 is present in colon tissue, where it may contribute to inflammation, whereas DP most likely promotes anti-inflammatory actions.
The Wnt/β-Catenin Pathway Attenuates Experimental Allergic Airway Disease
Authors: Reuter Sebastian , Martin Helen , Beckert Hendrik , Bros Matthias , Montermann Evelyn , Belz Christina , Heinz Anke , Ohngemach Svetlana , Sahin Ugur , Stassen Michael , Buhl Roland , Eshkind Leonid , Taube Christian
Published: June 2014

Signaling via the Wnt/β-catenin pathway plays crucial roles in embryogenesis and homeostasis of adult tissues. In the lung, the canonical Wnt/β-catenin pathway has been implicated in remodeling processes, development of emphysema, and fibrosis. However, its relevance for the modulation of allergic responses in the lung remains unclear. Using genetically modified mice with lung-specific inducible (doxycycline) Wnt-1 expression (CCSP-rtTA × tetO-Wnt1), the impact of Wnt on the development of allergic airway disease was analyzed. Overexpression of Wnt during the allergen challenge phase attenuated the development of airway inflammation in an acute model, as well as in a more therapeutic model of secondary challenge. These findings were further supported by treatment of allergen-sensitized mice with LiCl during challenge. Similar to Wnt, LiCl prevented the degradation of β-catenin and, thus, attenuated allergic airway inflammation and hyperresponsiveness. Migration studies revealed that lung-specific expression of Wnt reduced the migration of Ag-loaded dendritic cells (DCs) into the draining lymph nodes following allergen challenge. Administration of in vitro allergen-loaded DCs overcame Wnt-mediated suppression of airway inflammation. Furthermore, in vitro studies confirmed that DC-dependent T cell activation is impaired by blocking β-catenin degradation. These results demonstrate an important role for the canonical Wnt/β-catenin pathway in the DC-mediated regulation of allergic responses in the lung.
Rituximab Efficiently Depletes Increased CD20-Expressing T Cells in Multiple Sclerosis Patients
Authors: Palanichamy Arumugam , Jahn Sarah , Nickles Dorothee , Derstine Mia , Abounasr Aya , Hauser Stephen L. , Baranzini Sergio E. , Leppert David , von Büdingen H.-Christian
Published: June 2014

In multiple sclerosis (MS), B cell–depleting therapy using monoclonal anti-CD20 Abs, including rituximab (RTX) and ocrelizumab, effectively reduces disease activity. Based on indirect evidence, it is generally believed that elimination of the Ag-presenting capabilities and Ag nonspecific immune functions of B cells underlie the therapeutic efficacy. However, a small subset of T lymphocytes (T cells) was shown to also express CD20, but controversy prevails surrounding the true existence of this T cell subpopulation. Using single-cell imaging flow cytometry and expression profiling of sorted lymphocyte subsets, we unequivocally demonstrate the existence of CD3+CD20dim T cells. We show that in MS patients, increased levels of CD3+CD20dim T cells are effectively depleted by RTX. The pathological relevance of this T cell subset in MS remains to be determined. However, given their potential proinflammatory functionality, depletion of CD20-expressing T cells may also contribute to the therapeutic effect of RTX and other mAbs targeting CD20.