bluuebunny/crossref_metadata_2025_split huggingface dataset:
Macropinocytosis Is the Entry Mechanism of Amphotropic Murine Leukemia Virus
Authors: Rasmussen Izabela , Vilhardt Frederik 
Published: November 2014

ABSTRACT
The entry mechanism of murine amphotropic retrovirus (A-MLV) has not been unambiguously determined. We show here that A-MLV is internalized not by caveolae or other pinocytic mechanisms but by macropinocytosis. Thus, A-MLV infection of mouse embryonic fibroblasts deficient for caveolin or dynamin, and NIH 3T3 cells knocked down for caveolin expression, was unaffected. Conversely, A-MLV infection of NIH 3T3 and HeLa cells was sensitive to amiloride analogues and actin-depolymerizing drugs that interfere with macropinocytosis. Further manipulation of the actin cytoskeleton through conditional expression of dominant positive or negative mutants of Rac1, PAK1, and RhoG, to increase or decrease macropinocytosis, similarly correlated with an augmented or inhibited infection with A-MLV, respectively. The same experimental perturbations affected the infection of viruses that use clathrin-coated-pit endocytosis or other pathways for entry only mildly or not at all. These data agree with immunofluorescence studies and cryo-immunogold labeling for electron microscopy, which demonstrate the presence of A-MLV in protrusion-rich areas of the cell surface and in cortical fluid phase (dextran)-filled macropinosomes, which also account for up to a half of the cellular uptake of the cell surface-binding lectin concanavalin A. We conclude that A-MLV use macropinocytosis as the predominant entry portal into cells.

IMPORTANCE
Binding and entry of virus particles into mammalian cells are the first steps of infection. Understanding how pathogens and toxins exploit or divert endocytosis pathways has advanced our understanding of membrane trafficking pathways, which benefits development of new therapeutic schemes and methods of drug delivery. We show here that amphotropic murine leukemia virus (A-MLV) pseudotyped with the amphotropic envelope protein (which expands the host range to many mammalian cells) gains entry into host cells by macropinocytosis. Macropinosomes form as large, fluid-filled vacuoles (up to 10 μm) following the collapse of cell surface protrusions and membrane scission. We used drugs or the introduction of mutant proteins that affect the actin cytoskeleton and cell surface dynamics to show that macropinocytosis and A-MLV infection are correlated, and we provide both light- and electron-microscopic evidence to show the localization of A-MLV in macropinosomes. Finally, we specifically exclude some other potential entry portals, including caveolae, previously suggested to internalize A-MLV.
The Level of Viral Infection of Antigen-Presenting Cells Correlates with the Level of Development of Theiler's Murine Encephalomyelitis Virus-Induced Demyelinating Disease
Authors: Jin Young Hee , Kang Hyun Seok , Hou Wanqiu , Meng Liping , Kim Byung S. 
Published: November 2014

ABSTRACTIntracerebral infection with Theiler's murine encephalomyelitis virus (TMEV) induces immune-mediated demyelinating disease in susceptible SJL/J mice but not in resistant C57BL/6 mice. Previous studies have indicated that the major histocompatibility complex (MHC) genes play the most prominent role in the development of TMEV-induced demyelinating disease. In this study, we used C57BL/6.S (B6.S) congenic mice, which carryH-2sMHC genes instead ofH-2bMHC genes in conjunction with the C57BL/6 (B6) background genes. Our data show that virus-infected B6.S mice are free from disease and have significantly lower viral loads than susceptible SJL mice, particularly in the spinal cord. A strong protective Th1-type T helper response with virtually no pathogenic Th17 response was detected in B6.S mice, in contrast to the reduced Th1- and robust Th17-type responses in SJL mice. Notably, lower levels of viral infectivity in B6.S antigen-presenting cells (APCs) correlated with the disease resistance and T-cell-type response.In vitrostudies using APCs from B6.S and SJL mice show that TLR2, -3, -4, and -7, but not TLR9, signaling can replace viral infection and augment the effect of viral infection in the differentiation of the pathogenic Th17 cell type. Taken together, these results strongly suggest that the viral replication levels in APCs critically affect the induction of protective versus pathogenic Th cell types via the signaling of pattern recognition receptors for innate immune responses. Our current findings further imply that the levels of viral infectivity/replication and TLR-mediated signaling play critical roles in the pathogenesis of chronic viral diseases.IMPORTANCEThis study indicates that innate immune cytokines produced in antigen-presenting cells stimulating the T cell immune responses during early viral infection play a critical role in determining the susceptibility of mice to the development of demyelinating disease. The level of innate immune cytokines reflects the level of initial viral infection in the antigen-presenting cells, and the level determines the development of T cell types, which are either protective or pathogenic. The level of initial viral infection to the cells is controlled by a gene or genes that are not associated with the major histocompatibility antigen complex genes. This finding has an important implication in controlling not only chronic viral infections but also infection-induced autoimmune-like diseases, which are closely associated with the pathogenic type of T cell responses.
Vaccine-Induced Protection against Orthopoxvirus Infection Is Mediated through the Combined Functions of CD4 T Cell-Dependent Antibody and CD8 T Cell Responses
Authors: Chaudhri Geeta , Tahiliani Vikas , Eldi Preethi , Karupiah Gunasegaran 
Published: November 2014

ABSTRACTAntibody production by B cells in the absence of CD4 T cell help has been shown to be necessary and sufficient for protection against secondary orthopoxvirus (OPV) infections. This conclusion is based on short-term depletion of leukocyte subsets in vaccinated animals, in addition to passive transfer of immune serum to naive hosts that are subsequently protected from lethal orthopoxvirus infection. Here, we show that CD4 T cell help is necessary for neutralizing antibody production and virus control during a secondary ectromelia virus (ECTV) infection. A crucial role for CD4 T cells was revealed when depletion of this subset was extended beyond the acute phase of infection. Sustained depletion of CD4 T cells over several weeks in vaccinated animals during a secondary infection resulted in gradual diminution of B cell responses, including neutralizing antibody, contemporaneous with a corresponding increase in the viral load. Long-term elimination of CD8 T cells alone delayed virus clearance, but prolonged depletion of both CD4 and CD8 T cells resulted in death associated with uncontrolled virus replication. In the absence of CD4 T cells, perforin- and granzyme A- and B-dependent effector functions of CD8 T cells became critical. Our data therefore show that both CD4 T cell help for antibody production and CD8 T cell effector function are critical for protection against secondary OPV infection. These results are consistent with the notion that the effectiveness of the smallpox vaccine is related to its capacity to induce both B and T cell memory.IMPORTANCESmallpox eradication through vaccination is one of the most successful public health endeavors of modern medicine. The use of various orthopoxvirus (OPV) models to elucidate correlates of vaccine-induced protective immunity showed that antibody is critical for protection against secondary infection, whereas the role of T cells is unclear. Short-term leukocyte subset depletion in vaccinated animals or transfer of immune serum to naive, immunocompetent hosts indicates that antibody alone is necessary and sufficient for protection. We show here that long-term depletion of CD4 T cells over several weeks in vaccinated animals during secondary OPV challenge reveals an important role for CD4 T cell-dependent antibody responses in effective virus control. Prolonged elimination of CD8 T cells alone delayed virus clearance, but depletion of both T cell subsets resulted in death associated with uncontrolled virus replication. Thus, vaccinated individuals who subsequently acquire T cell deficiencies may not be protected against secondary OPV infection.
Receptor Recognition Mechanisms of Coronaviruses: a Decade of Structural Studies
Authors: Li Fang 
Published: November 2014

ABSTRACT
Receptor recognition by viruses is the first and essential step of viral infections of host cells. It is an important determinant of viral host range and cross-species infection and a primary target for antiviral intervention. Coronaviruses recognize a variety of host receptors, infect many hosts, and are health threats to humans and animals. The receptor-binding S1 subunit of coronavirus spike proteins contains two distinctive domains, the N-terminal domain (S1-NTD) and the C-terminal domain (S1-CTD), both of which can function as receptor-binding domains (RBDs). S1-NTDs and S1-CTDs from three major coronavirus genera recognize at least four protein receptors and three sugar receptors and demonstrate a complex receptor recognition pattern. For example, highly similar coronavirus S1-CTDs within the same genus can recognize different receptors, whereas very different coronavirus S1-CTDs from different genera can recognize the same receptor. Moreover, coronavirus S1-NTDs can recognize either protein or sugar receptors. Structural studies in the past decade have elucidated many of the puzzles associated with coronavirus-receptor interactions. This article reviews the latest knowledge on the receptor recognition mechanisms of coronaviruses and discusses how coronaviruses have evolved their complex receptor recognition pattern. It also summarizes important principles that govern receptor recognition by viruses in general.
Nonmuscle Myosin Heavy Chain IIB Mediates Herpes Simplex Virus 1 Entry
Authors: Arii Jun , Hirohata Yoshitaka , Kato Akihisa , Kawaguchi Yasushi 
Published: November 2014

ABSTRACT
Nonmuscle myosin heavy chain IIA (NMHC-IIA) has been reported to function as a herpes simplex virus 1 (HSV-1) entry coreceptor by interacting with viral envelope glycoprotein B (gB). Vertebrates have three genetically distinct isoforms of the NMHC-II, designated NMHC-IIA, NMHC-IIB, and NMHC-IIC. COS cells, which are readily infected by HSV-1, do not express NMHC-IIA but do express NMHC-IIB. This observation prompted us to investigate whether NMHC-IIB might associate with HSV-1 gB and be involved in an HSV-1 entry like NMHC-IIA. In these studies, we show that (i) NMHC-IIB coprecipitated with gB in COS-1 cells upon HSV-1 entry; (ii) a specific inhibitor of myosin light chain kinase inhibited cell surface expression of NMHC-IIB in COS-1 cells upon HSV-1 entry as well as HSV-1 infection, as reported with NMHC-IIA; (iii) overexpression of mouse NMHC-IIB in IC21 cells significantly increased their susceptibility to HSV-1 infection; and (iv) knockdown of NMHC-IIB in COS-1 cells inhibited HSV-1 infection as well as cell-cell fusion mediated by HSV-1 envelope glycoproteins. These results supported the hypothesis that, like NMHC-IIA, NMHC-IIB associated with HSV-1 gB and mediated HSV-1 entry.

IMPORTANCE
Herpes simplex virus 1 (HSV-1) was reported to utilize nonmuscle myosin heavy chain IIA (NMHC-IIA) as an entry coreceptor associating with gB. Vertebrates have three genetically distinct isoforms of NMHC-II. In these isoforms, NMHC-IIB is of special interest since it highly expresses in neuronal tissue, one of the most important cellular targets of HSV-1
in vivo
. In this study, we demonstrated that the ability to mediate HSV-1 entry appeared to be conserved in NMHC-II isoforms. These results may provide an insight into the mechanism by which HSV-1 infects a wide variety of cell types
in vivo
.
Factors associated with the age of the onset of diabetes in women aged 50 years or more: a population-based study
Authors: Valadares Ana L R , Machado Vanessa S S , Costa-Paiva Lúcia S , de Sousa Maria H , Pinto-Neto Aarão M 
Published: November 2014

ObjectiveInvestigate factors associated with the onset of diabetes in women aged more than 49 years.Design and methodsCross-sectional, population-based study using self-reports with 622 women. The dependent variable was the age of occurrence of diabetes using the life table method. Cox multiple regression models were adjusted to analyse the onset of diabetes according to predictor variables. Sociodemographic, clinical and behavioural factors were evaluated.ResultsOf the 622 women interviewed, 22.7% had diabetes. The mean age at onset was 56 years. The factors associated with the age of occurrence of diabetes were self-rated health (very good, good) (coefficient=−0.792; SE of the coefficient=0.215; p=0.0001), more than two individuals living in the household (coefficient=0.656, SE of the coefficient=0.223; p=0.003), and body mass index (BMI) (kg/m2) at 20–30 years of age (coefficient= 0.056, SE of the coefficient=0.023; p=0.014).ConclusionsSelf-rated health considered good or very good was associated with a higher rate of survival without diabetes. Sharing a home with two or more other people and a weight increase at 20–30 years of age was associated with the onset of type 2 diabetes.
A pilot study of a digital drainage system in pneumothorax
Authors: Tunnicliffe Georgia , Draper Adrian 
Published: December 2014

Over recent years there has been increasing usage of digital systems within cardiothoracic surgery to quantify air leaks and aid in clinical decision-making regarding the removal of chest drains postoperatively. The literature suggests improved agreement on timing of removal of chest drains and a reduced length of stay of patients. It could be that such devices could be useful tools for the clinician managing cases of pneumothorax.MethodsThis pilot study recruited adults admitted under the medical team with a pneumothorax requiring a chest drain. Participants had the underwater seal device changed for a digital device (Thopaz) which allowed continuous monitoring of the air leak. Drains were removed when either there was no ongoing air leak and the lung had expanded, or surgery was deemed necessary.ResultsThirteen patients with pneumothorax (four primary, nine secondary) used the device during their admission including one patient treated in the community (the device has internal suction). Data were used to aid the clinician in management of the pneumothorax including the timing of surgery/ removal of drain and commencement of suction.DiscussionDigital devices appear to be safe and effective and may prove to be a useful tool in the management of pneumothorax.
A case-crossover study of transient risk factors influence on occupational injuries: a study protocol based on a review of previous studies
Authors: Oesterlund Anna H , Lander Flemming , Lauritsen Jens 
Published: November 2014

BackgroundThe occupational injury incident rate remains relatively high in the European Union. The case-crossover study gives a unique opportunity to study transient risk factors that normally would be very difficult to approach. Studies like this have been carried out in both America and Asia, but so far no relevant research has been conducted in Europe.MethodCase-crossover studies of occupational injuries were collected from PubMed and Embase and read through. Previous experiences concerning method, exposure and outcome, time-related measurements and construction of the questionnaire were taken into account in the preparation of a pilot study. Consequently, experiences from the pilot study were used to design the study protocol. Approximately 2000 patients with an occupational injury will be recruited from the emergency departments in Herning and Odense, Denmark. A standardised questionnaire will be used to collect basic demographic data and information on eight transient risk factors. Based on previous studies and knowledge on occupational injuries the transient risk factors we chose to examine were: time pressure, performing a task with a different method/using unaccustomed technique, change in working surroundings, using a phone, disagreement, feeling ill, being distracted and using malfunctioning machinery/tools or work material. Exposure time ‘just before the injury’ will be compared with two control periods, ‘previous day at the same time of the injury’ (pair match) and the previous work week (usual frequency).DiscussionThis study protocol describes a unique opportunity to calculate the effect of transient risk factors on occupational injuries in a European setting.
Patient-reported outcome measures for patients with cerebral aneurysms acquired via social media: data from a large nationwide sample
Authors: Chen Michael , Mangubat Erwin , Ouyang Bichun 
Published: December 2014

BackgroundWith greater survival rates, patient-reported outcome measures (PROMs) among survivors of ruptured cerebral aneurysm should be an increasing concern among neurointerventionalists. Prior studies were limited in scale and generalizability. Our study aims were to (1) evaluate the validity of cerebral aneurysm PROMs obtained from social media; (2) determine the persistence of PROMs over time; and (3) determine what PROMs still exist in those with no physical impairments.MethodsBy engaging national brain aneurysm support groups and using an online questionnaire modeled after the generic EQ-5D instrument, we asked respondents to classify their health in five dimensions including mobility, self-care, usual activities, pain/discomfort, and anxiety/depression using a 3-point Likert scale.ResultsIn 2 months we received 604 responses from 46 states in the USA. Our cohort of ruptured aneurysm respondents reported PROMs similar to previously published series. Over time, headache and anxiety improved while depression, level of exercise, and return to work remained unchanged. We found that memory worsened after 2 years. Among those without any physical impairment, rates of 20.6%, 14.9%, 12.6%, and 23% were seen for significant headaches, significant memory loss, significant depression, and sense of life being negatively affected, respectively.ConclusionsDespite this novel study design, we obtained results comparable to prior studies. These results suggest that many patients with ruptured cerebral aneurysms, regardless of whether they are >2 years after the event and/or free of physical impairment, struggle with a poor quality of life. The latency, scale, and low cost of this study design may accelerate future cerebral aneurysm PROM research.
There is no fork
Authors: Marlow Simon , Brandy Louis , Coens Jonathan , Purdy Jon 
Published: December 2014

We describe a new programming idiom for concurrency, based on Applicative Functors, where concurrency is implicit in the Applicative <*> operator. The result is that concurrent programs can be written in a natural applicative style, and they retain a high degree of clarity and modularity while executing with maximal concurrency. This idiom is particularly useful for programming against external data sources, where the application code is written without the use of explicit concurrency constructs, while the implementation is able to batch together multiple requests for data from the same source, and fetch data from multiple sources concurrently. Our abstraction uses a cache to ensure that multiple requests for the same data return the same result, which frees the programmer from having to arrange to fetch data only once, which in turn leads to greater modularity.
While it is generally applicable, our technique was designed with a particular application in mind: an internal service at Facebook that identifies particular types of content and takes actions based on it. Our application has a large body of business logic that fetches data from several different external sources. The framework described in this paper enables the business logic to execute efficiently by automatically fetching data concurrently; we present some preliminary results.
Using formal methods to enable more secure vehicles
Authors: Fisher Kathleen 
Published: December 2014

Networked embedded systems are ubiquitous in modern society. Examples include SCADA systems that manage physical infrastructure, medical devices such as pacemakers and insulin pumps, and vehicles such as airplanes and automobiles. Such devices are connected to networks for a variety of compelling reasons, including the ability to access diagnostic information conveniently, perform software updates, provide innovative features, and lower costs. Researchers and hackers have shown that these kinds of networked embedded systems are vulnerable to remote attacks and that such attacks can cause physical damage and can be hidden from monitors [1, 4].
DARPA launched the HACMS program to create technology to make such systems dramatically harder to attack successfully. Specifically, HACMS is pursuing a clean-slate, formal methods-based approach to the creation of high-assurance vehicles, where high assurance is defined to mean functionally correct and satisfying appropriate safety and security properties. Specific technologies include program synthesis, domain-specific languages, and theorem provers used as program development environments. Targeted software includes operating system components such as hypervisors, microkernels, file systems, and device drivers as well as control systems such as autopilots and adaptive cruise controls. Program researchers are leveraging existing high-assurance software including NICTA's seL4 microkernel and INRIA's CompCert compiler.
Although the HACMS project is less than halfway done, the program has already achieved some remarkable success. At program kick-off, a Red Team easily hijacked the baseline open-source quadcopter that HACMS researchers are using as a research platform. At the end of eighteen months, the Red Team was not able to hijack the newly-minted "SMACCMCopter" running high-assurance HACMS code, despite being given six weeks and full access to the source code of the copter. An expert in penetration testing called the SMACCMCopter "the most secure UAV on the planet."
In this talk, I will describe the HACMS program: its motivation, the underlying technologies, current results, and future directions.