bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human regulatory macrophages are potent in suppression of the xenoimmune response via indoleamine‐2,3‐dioxygenase‐involved mechanism(s)
Authors: Guo Fei , Hu Min , Huang Dandan , Zhao Yuanfei , Heng Benjamin , Guillemin Gilles , Lim Chai K. , Hawthorne Wayne J. , Yi Shounan 
Published: August 2017

AbstractBackgroundFor xenotransplantation to truly succeed, we must develop immunomodulatory strategies to suppress the xenoimmune response but by minimizing immunosuppression over the long term. Regulatory macrophages (Mreg) have been shown to suppress polyclonal T‐cell proliferation in vitro and prolong allograft survival in vivo. However, the question of whether they are capable of suppressing xenoimmune responses remains unknown. This study assessed the potential of human Mreg to be used as an effective immunomodulatory method in xenotransplantation.MethodsCD14+ monocytes selected from human peripheral blood mononuclear cells (PBMC) were cultured with macrophage colony‐stimulating factor (M‐CSF) for 7 days with IFN‐γ added at day 6 for Mreg induction. Mreg phenotyping was performed by flow cytometric analysis, and the in vitro suppressive function was assessed by mixed lymphocyte reaction (MLR) using irradiated pig PBMC as the xenogeneic stimulator cells, human PBMC as responder cells, and autologous Mreg as suppressor cells. To assess mRNA expression of Mreg functional molecules indoleamine‐2,3‐dioxygenase (IDO), IL‐10, inducible nitric oxide synthase (iNOS) and TGF‐β were measured by real‐time PCR. Supernatants were collected from the MLR cultures for IDO activity assay by high‐performance liquid chromatography (HPLC). The effects of the IDO inhibitor 1‐D/L‐methyl‐tryptophan (1‐MT), iNOS inhibitor NG‐monomethyl‐l‐arginine (L‐NMMA), and anti‐IFN‐γ or anti‐TGF‐β monoclonal antibody (mAb) treatment on Mreg suppressive capacity were tested from the supernatants of the MLR assays.ResultsWe demonstrated that induced Mreg with a phenotype of CD14lowCD16−/lowCD80lowCD83−/lowCD86+/hiHLA‐DR+/hi were capable of suppressing proliferating human PBMC, CD4+, and CD8+ T cells, even at a higher responder:Mreg ratio of 32:1 in a pig‐human xenogeneic MLR. The strong suppressive potency of Mreg was further correlated with their upregulated IDO expression and activity. The IDO upregulation of Mreg was associated with an increased production of IFN‐γ, an IDO stimulator, by xenoreactive responder cells in the xenogeneic MLR. While no effect on Mreg suppressive potency was detected by addition of the iNOS inhibitor L‐NMMA or anti‐TGF‐β mAb into the MLR assays, inhibition of IDO activity by neutralizing IFN‐γ or by IDO inhibitor 1‐MT substantially impaired the capacity of Mreg to suppress the xenogeneic response, indicating the importance of upregulated IDO activity in Mreg‐mediated suppression of the xenogeneic response in vitro.ConclusionThis study demonstrates that human Mreg are capable of suppressing the xenoimmune response in vitro via IDO‐involved mechanism(s), suggesting their potential role as an effective immunomodulatory tool in xenotransplantation.
Experimental and modelling evidence of shortening heat in cardiac muscle
Authors: Tran Kenneth , Han June‐Chiew , Crampin Edmund John , Taberner Andrew James , Loiselle Denis Scott 
Published: August 2017

Key points
Heat associated with muscle shortening has been repeatedly demonstrated in skeletal muscle, but its existence in cardiac muscle remains contentious after five decades of study.
By iterating between experiments and computational modelling, we show compelling evidence for the existence of shortening heat in cardiac muscle and reveal, mechanistically, the source of this excess heat.
Our results clarify a long‐standing uncertainty in the field of cardiac muscle energetics.
We provide a revised partitioning of cardiac muscle energy expenditure to include this newly revealed thermal component.
AbstractWhen a muscle shortens against an afterload, the heat that it liberates is greater than that produced by the same muscle contracting isometrically at the same level of force. This excess heat is defined as ‘shortening heat’, and has been repeatedly demonstrated in skeletal muscle but not in cardiac muscle. Given the micro‐structural similarities between these two muscle types, and since we imagine that shortening heat is the thermal accompaniment of cross‐bridge cycling, we have re‐examined this issue. Using our flow‐through microcalorimeter, we measured force and heat generated by isolated rat trabeculae undergoing isometric contractions at different muscle lengths and work‐loop (shortening) contractions at different afterloads. We simulated these experimental protocols using a thermodynamically constrained model of cross‐bridge cycling and probed the mechanisms underpinning shortening heat. Predictions generated by the model were subsequently validated by a further set of experiments. Both our experimental and modelling results show convincing evidence for the existence of shortening heat in cardiac muscle. Its magnitude is inversely related to the afterload or, equivalently, directly related to the extent of shortening. Computational simulations reveal that the heat of shortening arises from the cycling of cross‐bridges, and that the rate of ATP hydrolysis is more sensitive to change of muscle length than to change of afterload. Our results clarify a long‐standing uncertainty in the field of cardiac muscle energetics.
Are Measures Derived From Land Use and Transport Policies Associated With Walking for Transport?
Authors: Rachele Jerome N. , Learnihan Vincent , Badland Hannah M. , Mavoa Suzanne , Turrell Gavin , Giles-Corti Billie 
Published: August 2017

Background: There is growing urgency for higher quality evidence to inform policy. This study developed geographic information system spatial measures based on land use and transport policies currently used in selected Australian states to assess which, if any, of these measures were associated with walking for transport. Methods: Overall, 6901 participants from 570 neighborhoods in Brisbane, Australia, were included. Participants reported their minutes of walking for transport in the previous week. After a review of state-level land use and transport policies relevant to walking for transport across Australia, 7 geographic information system measures were developed and tested based on 9 relevant policies. Data were analyzed using multilevel multinomial logistic regression. Results: Greater levels of walking for transport were associated with more highly connected street networks, the presence of public transport stops, and having at least 2 public transport services per hour. Conversely, neighborhoods with shorter cul-de-sac lengths had lower levels of walking for transport. There was no evidence of associations between walking for transport and street block lengths less than 240 m or traffic volumes. Conclusions: These findings highlight the need for urban design and transport policies developed by governments to be assessed for their impact on transport-related physical activity.
Teacher Development in First-Year Physical Educators: A Comparison of Effectiveness Among Different Physical Education Teacher Education Backgrounds
Authors: Ensign Julene , Woods Amelia Mays , Kulinna Pamela Hodges 
Published: August 2017

Purpose:This study evaluated the teaching effectiveness of six first-year physical educators, three Southwestern and three Midwestern graduates, employing different curricular approaches.Method:Utilizing surveys, interviews, questionnaires, and systematic observations, data were analyzed through a framework of seven essential teaching tasks (Rink, 2002).Results:Data indicated overall mean scores of 34% motor appropriate activity with Academic Learning Time-Physical Education (ALT-PE) and a rating of 70.37 on the Qualitative Measures of Teacher Performance Scale (QMTPS). Notable contrasts included higher mean scores for Southwest participants for motor-appropriate and motor-inappropriate activity. Midwest participants devoted more time to game situations, management, and social behavior. For QMTPS, Southwest means were higher in every category. Qualitative themes produced similarities in teaching philosophy, fidelity to preservice training, and perceived value of reflective practices. Contrasts existed in curricular emphases and approaches to classroom management.Discussion/Conclusion:Characteristics of effective teaching were demonstrated by all participants regardless of curricular emphasis.
Improved Serodiagnostic Performance for Lyme Disease by Use of Two Recombinant Proteins in Enzyme-Linked Immunosorbent Assay Compared to Standardized Two-Tier Testing
Authors: Bradshaw Gary L. , Thueson R. Kelley , Uriona Todd J. 
Published: August 2017

ABSTRACT

The most reliable test method for the serological confirmation of Lyme disease (LD) is a 2-tier method recommended by the CDC in 1995. The first-tier test is a low-specificity enzyme-linked immunosorbent assay (ELISA), and the second-tier tests are higher-specificity IgG and IgM Western blots. This study describes the selection of two
Borrelia burgdorferi
recombinant proteins and evaluation of their performance in a simple 1-tier test for the serological confirmation of LD. These two proteins were generated from (i) the full-length
dbpA
gene combined with the invariable region 6 of the
vlsE
gene (DbpA/C6) and (b) the full-length
ospC
gene (OspC). The expressed DbpA/C6 and OspC proteins were useful in detecting anti-
Borrelia
IgG and IgM antibodies, respectively. A blind study was conducted on a well-characterized panel of 279 human sera from the CDC, comparing ELISAs using these two recombinant antigens with the 2-tier test method. The two methods (DbpA/C6-OspC versus 2-tier test) were equivalent in identifying sera from negative-control subjects (99% and 100% specificity, respectively) and in detecting stage II and III LD patient sera (100% and 100% sensitivity). However, the DbpA/C6-OspC ELISA was markedly better (80% versus 63%) than the 2-tier test method in detecting anti-
Borrelia
antibodies in stage I LD patients. The findings suggest that these antigens could be used in a simple 1-tier ELISA that is faster to perform, easier to interpret, and less expensive than the 2-tier test method and which is better at detecting
Borrelia
-specific antibodies in sera from patients with stage I LD.
Replication of JC Virus DNA in the G144 Oligodendrocyte Cell Line Is Dependent Upon Akt
Authors: Peterson Jesse N. , Lin Brian , Shin Jong , Phelan Paul J. , Tsichlis Philip , Schwob James E. , Bullock Peter A. 
Published: August 2017

ABSTRACT
Progressive multifocal leukoencephalopathy (PML) is an often-fatal demyelinating disease of the central nervous system. PML results when oligodendrocytes within immunocompromised individuals are infected with the human JC virus (JCV). We have identified an oligodendrocyte precursor cell line, termed G144, that supports robust levels of JCV DNA replication, a central part of the JCV life cycle. In addition, we have determined that JC virus readily infects G144 cells. Furthermore, we have determined that JCV DNA replication in G144 cells is stimulated by myristoylated (i.e., constitutively active) Akt and reduced by the Akt-specific inhibitor MK2206. Thus, this oligodendrocyte-based model system will be useful for a number of purposes, such as studies of JCV infection, establishing key pathways needed for the regulation of JCV DNA replication, and identifying inhibitors of this process.

IMPORTANCE
The disease progressive multifocal leukoencephalopathy (PML) is caused by the infection of particular brain cells, termed oligodendrocytes, by the JC virus. Studies of PML, however, have been hampered by the lack of an immortalized human cell line derived from oligodendrocytes. Here, we report that the G144 oligodendrocyte cell line supports both infection by JC virus and robust levels of JCV DNA replication. Moreover, we have established that the Akt pathway regulates JCV DNA replication and that JCV DNA replication can be inhibited by MK2206, a compound that is specific for Akt. These and related findings suggest that we have established a powerful oligodendrocyte-based model system for studies of JCV-dependent PML.
Ubiquitin Ligase WWP1 Interacts with Ebola Virus VP40 To Regulate Egress
Authors: Han Ziying , Sagum Cari A. , Takizawa Fumio , Ruthel Gordon , Berry Corbett T. , Kong Jing , Sunyer J. Oriol , Freedman Bruce D. , Bedford Mark T. , Sidhu Sachdev S. , Sudol Marius , Harty Ronald N. 
Published: August 2017

ABSTRACT

Ebola virus (EBOV) is a member of the
Filoviridae
family and the cause of hemorrhagic fever outbreaks. The EBOV VP40 (eVP40) matrix protein is the main driving force for virion assembly and budding. Indeed, expression of eVP40 alone in mammalian cells results in the formation and budding of virus-like particles (VLPs) which mimic the budding process and morphology of authentic, infectious EBOV. To complete the budding process, eVP40 utilizes its PPXY L-domain motif to recruit a specific subset of host proteins containing one or more modular WW domains that then function to facilitate efficient production and release of eVP40 VLPs. In this report, we identified additional host WW-domain interactors by screening for potential interactions between mammalian proteins possessing one or more WW domains and WT or PPXY mutant peptides of eVP40. We identified the HECT family E3 ubiquitin ligase WWP1 and all four of its WW domains as strong interactors with the PPXY motif of eVP40. The eVP40-WWP1 interaction was confirmed by both peptide pulldown and coimmunoprecipitation assays, which also demonstrated that modular WW domain 1 of WWP1 was most critical for binding to eVP40. Importantly, the eVP40-WWP1 interaction was found to be biologically relevant for VLP budding since (i) small interfering RNA (siRNA) knockdown of endogenous WWP1 resulted in inhibition of eVP40 VLP egress, (ii) coexpression of WWP1 and eVP40 resulted in ubiquitination of eVP40 and a subsequent increase in eVP40 VLP egress, and (iii) an enzymatically inactive mutant of WWP1 (C890A) did not ubiquitinate eVP40 or enhance eVP40 VLP egress. Last, our data show that ubiquitination of eVP40 by WWP1 enhances egress of VLPs and concomitantly decreases cellular levels of higher-molecular-weight oligomers of eVP40. In sum, these findings contribute to our fundamental understanding of the functional interplay between host E3 ligases, ubiquitination, and regulation of EBOV VP40-mediated egress.

IMPORTANCE
Ebola virus (EBOV) is a high-priority, emerging human pathogen that can cause severe outbreaks of hemorrhagic fever with high mortality rates. As there are currently no approved vaccines or treatments for EBOV, a better understanding of the biology and functions of EBOV-host interactions that promote or inhibit viral budding is warranted. Here, we describe a physical and functional interaction between EBOV VP40 (eVP40) and WWP1, a host E3 ubiquitin ligase that ubiquitinates VP40 and regulates VLP egress. This viral PPXY-host WW domain-mediated interaction represents a potential new target for host-oriented inhibitors of EBOV egress.
High-Risk Alphapapillomavirus Oncogenes Impair the Homologous Recombination Pathway
Authors: Wallace Nicholas A. , Khanal Sujita , Robinson Kristin L. , Wendel Sebastian O. , Messer Joshua J. , Galloway Denise A. 
Published: August 2017

ABSTRACT

Persistent high-risk genus human
Alphapapillomavirus
(HPV) infections cause nearly every cervical carcinoma and a subset of tumors in the oropharyngeal tract. During the decades required for HPV-associated tumorigenesis, the cellular genome becomes significantly destabilized. Our analysis of cervical tumors from four separate data sets found a significant upregulation of the homologous-recombination (HR) pathway genes. The increased abundance of HR proteins can be replicated in primary cells by expression of the two HPV oncogenes (E6 and E7) required for HPV-associated transformation. HPV E6 and E7 also enhanced the ability of HR proteins to form repair foci, and yet both E6 and E7 reduce the ability of the HR pathway to complete double-strand break (DSB) repair by about 50%. The HPV oncogenes hinder HR by allowing the process to begin at points in the cell cycle when the lack of a sister chromatid to serve as a homologous template prevents completion of the repair. Further, HPV E6 attenuates repair by causing RAD51 to be mislocalized away from both transient and persistent DSBs, whereas HPV E7 is only capable of impairing RAD51 localization to transient lesions. Finally, we show that the inability to robustly repair DSBs causes some of these lesions to be more persistent, a phenotype that correlates with increased integration of episomal DNA. Together, these data support our hypothesis that HPV oncogenes contribute to the genomic instability observed in HPV-associated malignancies by attenuating the repair of damaged DNA.

IMPORTANCE
This study expands the understanding of HPV biology, establishing a direct role for both HPV E6 and E7 in the destabilization of the host genome by blocking the homologous repair of DSBs. To our knowledge, this is the first time that both viral oncogenes were shown to disrupt this DSB repair pathway. We show that HPV E6 and E7 allow HR to initiate at an inappropriate part of the cell cycle. The mislocalization of RAD51 away from DSBs in cells expressing HPV E6 and E7 hinders HR through a distinct mechanism. These observations have broad implications. The impairment of HR by HPV oncogenes may be targeted for treatment of HPV
+
malignancies. Further, this attenuation of repair suggests HPV oncogenes may contribute to tumorigenesis by promoting the integration of the HPV genome, a common feature of HPV-transformed cells. Our data support this idea since HPV E6 stimulates the integration of episomes.
Preschool children who are frequent attenders in emergency departments: an observational study of associated demographics and clinical characteristics
Authors: Blair Mitch , Poots Alan J , Lim Valencia , Hiles Stephen , Greenfield Geva , Crehan Caroline , Kugler Beverley , Boreham Charlotte 
Published: August 2017

BackgroundUnscheduled visits to emergency departments (ED) have increased in the UK in recent years. Children who are repeat attenders are relatively understudied.AimsTo describe the sociodemographic and clinical characteristics of preschoolers who attend ED a large District General Hospital.Method/study designObservational study analysing routinely collected ED operational data. Children attending four or more visits per year were considered as ‘frequent attenders’. Poisson regression was used with demographic details (age, sex, ethnicity, sociodemographic status) to predict number of attendances seen in the year. We further analysed detailed diagnostic characteristics of a random sample of 10% of attendees.Main findings10 169 patients visited in the 12-month period with 16 603 attendances. 655 individuals attended on 3335 occasions. 6.4% of this population accounted for 20.1% of total visits. In the 10% sample, there were 304 attendances, and 69 (23%) had an underlying chronic long-standing illness (CLSI). This group were 2.4 times more likely to be admitted as inpatients compared with those without such conditions, median length of stay of 6.2 hours versus 2.5 hours (p=NS).ConclusionsFrequent ED attenders fall broadly into two distinct clinical groups: those who habitually return with self-limiting conditions and those with or without exacerbation of underlying CLSI. Both groups may be amenable to both additional nursing and other forms of community support to enhance self-care and continuity of care. Further research is required to increase our understanding of specific individual family and health system factors that predict repeat attendance in this age group.
Tailored lamellar keratoplasty: a new technique
Authors: Solaimani Mohammad , Tabatabaei Seyed Ali , Fallah Tafti Zahra 
Published: August 2017

We present a new method for treating complicated corneal gaps among patients who have previously undergone radial keratotomy (RK).After complete healing of a fungal keratitis in a patient who has undergone RK, we removed the oedematous corneal sector and put anterior tailored segment of a corneal button previously used for Descemet stripping endothelial keratoplasty. The patient's best-corrected visual acuity reached to 20/20 6 months later. This technique will compensate for oedematous parts without putting a significant effect on other parts such as purse string sutures.
Missed aortic valve endocarditis resulting in complete atrioventricular block and redo mechanical valve replacement
Authors: Harky Amer , Garner Megan , Popa Miruna , Shipolini Alex 
Published: August 2017

Infective endocarditis is a rare disease associated with high morbidity and mortality. As a result, early diagnosis and prompt antibiotic treatment with or without surgical intervention is crucial in the management of such condition.We report a case of missed infective endocarditis of the aortic valve. The patient underwent mechanical aortic valve replacement, with the native valve being sent for histopathological examination. On re-admission 16 months later, he presented with syncope, shortness of breathing and complete heart block. On review of the histopathology of native aortic valve, endocarditis was identified which had not been acted on. The patient underwent redo aortic valve replacement for severe aortic regurgitation.We highlight the importance of following up histopathological results as well as the need for multidisciplinary treatment of endocarditis with a combination of surgical and antibiotic therapy.
Randomised clinical simulation designed to evaluate the effect of telemedicine using Google Glass on cardiopulmonary resuscitation (CPR)
Authors: Pérez Alonso Nuria , Pardo Rios Manuel , Juguera Rodriguez Laura , Vera Catalan Tomas , Segura Melgarejo Francisca , Lopez Ayuso Belen , Martí­nez Riquelme Carolina , Lasheras Velasco Joaquin 
Published: August 2017

AimThrough a clinical simulation, this study aims to assess the effect of telematics support through Google Glass (GG) from an expert physician on performance of cardiopulmonary resuscitation (CPR) performed by a group of nurses, as compared with a control group of nurses receiving no assistance.MethodsThis was a randomised study carried out at the Catholic University of Murcia (November 2014–February 2015). Nursing professionals from the Emergency Medical Services in Murcia (Spain) were asked to perform in a clinical simulation of cardiac arrest. Half of the nurses were randomly chosen to receive coaching from physicians through GG, while the other half did not receive any coaching (controls). The main outcome of the study expected was successful defibrillation, which restores sinus rhythm.ResultsThirty-six nurses were enrolled in each study group. Statistically significant differences were found in the percentages of successful defibrillation (100% GG vs 78% control; p=0005) and CPR completion times: 213.91 s for GG and 250.31 s for control (average difference=36.39 s (95% CI 12.03 to 60.75), p=0.004).ConclusionsTelematics support by an expert through GG improves success rates and completion times while performing CPR in simulated clinical situations for nurses in simulated scenarios.