bluuebunny/crossref_metadata_2025 huggingface dataset:
Glycated Collagen Decreased Endothelial Cell Fibronectin Alignment in Response to Cyclic Stretch Via Interruption of Actin Alignment
Authors: Figueroa Dannielle S. , Kemeny Steven F. , Clyne Alisa Morss
Published: July 2014

Hyperglycemia is a defining characteristic of diabetes, and uncontrolled blood glucose in diabetes is associated with accelerated cardiovascular disease. Chronic hyperglycemia glycates extracellular matrix (ECM) collagen, which can lead to endothelial cell dysfunction. In healthy conditions, endothelial cells respond to mechanical stimuli such as cyclic stretch (CS) by aligning their actin cytoskeleton. Other cell types, specifically fibroblasts, align their ECM in response to CS. We previously demonstrated that glycated collagen inhibits endothelial cell actin alignment in response to CS. The aim of this study was to determine the effect of glycated collagen on ECM remodeling and protein alignment in response to stretch. Porcine aortic endothelial cells (PAEC) seeded on native or glycated collagen coated elastic substrates were exposed to 10% CS. Cells on native collagen aligned subcellular fibronectin fibers in response to stretch, whereas cells on glycated collagen did not. The loss of fibronectin alignment was due to inhibited actin alignment in response to CS, since fibronectin alignment did not occur in cells on native collagen when actin alignment was inhibited with cytochalasin. Further, while ECM protein content did not change in cells on native or glycated collagen in response to CS, degradation activity decreased in cells on glycated collagen. Matrix metalloproteinase 2 (MMP-2) and membrane-associated type 1 matrix metalloproteinase (MT1-MMP) protein levels decreased, and therefore MMP-2 activity also decreased. These MMP changes may relate to c-Jun N-terminal kinase (Jnk) phosphorylation inhibition with CS, which has previously been linked to focal adhesion kinase (FAK). These data demonstrate the importance of endothelial cell actin tension in remodeling and aligning matrix proteins in response to mechanical stimuli, which is critical to vascular remodeling in health and disease.
Surface Stress Effects on the Yield Strength in Nanotwinned Polycrystal Face-Centered-Cubic Metallic Nanowires
Authors: Zhu Linli , Guo Xiang , Lu Jian
Published: July 2014

The influence of surface stress on the yield strength of nanotwinned polycrystal face-centered-cubic (FCC) metallic nanowire is theoretically investigated. The contribution of surface boundaries on the strengthening/softening is analyzed in the framework of continuum mechanics theory by accounting for the surface energy effects. The other strengthening mechanisms originated from the inner boundaries are described by the Taylor model for the nanotwinned polycrystalline metals. The theoretical results demonstrate that the yield strength of nanotwinned polycrystal wires is dependent on the twin spacing, grain size and the geometrical size of the wire. The surface stress effects on the strength perform more and more significantly with decreasing the wire diameter, especially for the diameter smaller than 20 nm. In addition, the dependence of surface stress on the strength is also relevant to the size of microstructures as well as the magnitude and direction of surface stress. These results may be useful in evaluating the size-dependent mechanical performance of nanostructured materials.
On the Burst Strength Capacity of an Aging Subsea Gas Pipeline
Authors: Bin Mohd Mohd Hairil , Woo Kim Dong , Joon Lee Byeong , Kim Do Kyun , Kwan Seo Jung , Kee Paik Jeom
Published: July 2014

Precise evaluation of the performance of aging structures is particularly essential in the oil and gas industry, where inaccurate predictions of structural performance may lead to significantly hazardous consequences. It is also important to accurately predict the corrosion behavior of pipeline structures used in the production of gas in subsea areas. The effects of pipeline failure due to a significant reduction in burst strength make it hard for pipeline operators to maintain pipeline serviceability. Therefore, the serviceability of ongoing subsea gas pipelines should be assessed according to their burst strength capacities, which should not exceed the maximum allowable operating pressure (MAOP). In this study, the critical part of the corrosion along a 2.4 km pipeline was evaluated using two approaches: an empirical design codes formula and Ansys numerical analysis. The future integrity of the aging pipeline was then assessed to predict its remaining years of service. The results and outcomes of this study will be useful in evaluating pipeline integrity and predicting the remaining service life of aging pipeline structures.
Propylene Carbonate - XPS Reference Spectra
Authors: Stickle William , Stickle Tristan
Published: July 2014

We report the characterization of a thin film of propylene carbonate acquired at −120 °C deposited onto an aluminum substrate using a cold stage. Reference spectra for propylene carbonate were acquired using monochromatic Al Kα radiation. An XPS survey scan, a C 1s narrow scan, an O 1s narrow scan, an O KLL Auger narrow scan, an O 2s narrow scan and a valence band narrow scan were collected.
Novel, full 3D scintillation dosimetry using a static plenoptic camera
Authors: Goulet Mathieu , Rilling Madison , Gingras Luc , Beddar Sam , Beaulieu Luc , Archambault Louis
Published: July 2014

Purpose:

Patient‐specific quality assurance (QA) of dynamic radiotherapy delivery would gain from being performed using a 3D dosimeter. However, 3D dosimeters, such as gels, have many disadvantages limiting to quality assurance, such as tedious read‐out procedures and poor reproducibility. The purpose of this work is to develop and validate a novel type of high resolution 3D dosimeter based on the real‐time light acquisition of a plastic scintillator volume using a plenoptic camera. This dosimeter would allow for the QA of dynamic radiation therapy techniques such as intensity‐modulated radiation therapy (IMRT) or volumetric‐modulated arc therapy (VMAT).

Methods:

A Raytrix R5 plenoptic camera was used to image a 10 × 10 × 10 cm3 EJ‐260 plastic scintillator embedded inside an acrylic phantom at a rate of one acquisition per second. The scintillator volume was irradiated with both an IMRT and VMAT treatment plan on a Clinac iX linear accelerator. The 3D light distribution emitted by the scintillator volume was reconstructed at a 2 mm resolution in all dimensions by back‐projecting the light collected by each pixel of the light‐field camera using an iterative reconstruction algorithm. The latter was constrained by a beam's eye view projection of the incident dose acquired using the portal imager integrated with the linac and by physical consideration of the dose behavior as a function of depth in the phantom.

Results:

The absolute dose difference between the reconstructed 3D dose and the expected dose calculated using the treatment planning software Pinnacle3 was on average below 1.5% of the maximum dose for both integrated IMRT and VMAT deliveries, and below 3% for each individual IMRT incidences. Dose agreement between the reconstructed 3D dose and a radiochromic film acquisition in the same experimental phantom was on average within 2.1% and 1.2% of the maximum recorded dose for the IMRT and VMAT delivery, respectively.

Conclusions:

Using plenoptic camera technology, the authors were able to perform millimeter resolution, water‐equivalent dosimetry of an IMRT and VMAT plan over a whole 3D volume. Since no moving parts are required in the dosimeter, the incident dose distribution can be acquired as a function of time, thus enabling the validation of static and dynamic radiation delivery with photons, electrons, and heavier ions.
On the validity of density overrides for VMAT lung SBRT planning
Authors: Wiant David , Vanderstraeten Caroline , Maurer Jacqueline , Pursley Jan , Terrell Jonathon , Sintay Benjamin J.
Published: July 2014

Purpose:

Modeling dose to a moving target in lung is a very difficult task. Current approaches to planning lung stereotactic body radiotherapy (SBRT) generally calculate dose on either free breathing or average computed tomography (CT) scans, which do not always accurately predict dose to parts of the target volume not occupied by tumor on the planning scan. In this work, the authors look at using density overrides of the target volumes to more accurately predict dose for lung SBRT using the analytic anisotropic algorithm (AAA).

Methods:

Volumetric modulated arc therapy plans were created on free breathing scans (FBP), time average scans (AVGP), free breathing scans with the internal target volume overridden to tumor density (ITVP), free breathing scans with the planning target volume overridden to tumor density (PTVP), and free breathing scan using a hybrid scheme with the internal target volume set to tumor density and the planning target volume minus the internal target volume set to a density intermediate between lung and tumor (HP) for the case of a 4D motion phantom and five patient cases. Radiochromic film measurements were made for the phantom plans, with gamma analysis used to compare the planned to delivered dose. The patient plans were recalculated on each of the phases of a 4DCT to evaluate tumor coverage and conformity index (CI). A modified modulation complexity score (MCSv) and average open area per control point (AA) metrics were used to evaluate multileaf collimator (MLC) modulation for each of the plans.

Results:

The HP plans showed significantly higher gamma passing rates (p < 0.05) than the FBP, AVGP, and ITVP for criteria of 2 mm/2% and 1 mm/1%. No significant correlation was observed between gamma values and AA or MCSv. The tumor volume was covered by the prescription dose on all phases of the 4DCT for all patient plans. The PTVP and HP yielded lower mean CI than the other plans for all five patients, with three of the cases showing statistically significant differences (p < 0.05). No meaningful correlation was observed between the mean CI and AA or MCSv.

Conclusions:

These measurements suggest that the HP planning method may provide more accurate dose modeling and decreased normal lung irradiation for lung SBRT compared to the commonly used FBP and AVG planning methods when used with the AAA. The HP method does not appear to have a strong relationship with MLC modulation.
Genome interplay in the grain transcriptome of hexaploid bread wheat
Authors: Pfeifer Matthias , Kugler Karl G. , Sandve Simen R. , Zhan Bujie , Rudi Heidi , Hvidsten Torgeir R. ,   International Wheat Genome Sequencing Consortium, Mayer Klaus F. X. , Olsen Odd-Arne
Published: July 2014

Allohexaploid bread wheat (
Triticum aestivum
L.) provides approximately 20% of calories consumed by humans. Lack of genome sequence for the three homeologous and highly similar bread wheat genomes (A, B, and D) has impeded expression analysis of the grain transcriptome. We used previously unknown genome information to analyze the cell type–specific expression of homeologous genes in the developing wheat grain and identified distinct co-expression clusters reflecting the spatiotemporal progression during endosperm development. We observed no global but cell type– and stage-dependent genome dominance, organization of the wheat genome into transcriptionally active chromosomal regions, and asymmetric expression in gene families related to baking quality. Our findings give insight into the transcriptional dynamics and genome interplay among individual grain cell types in a polyploid cereal genome.
The future lies in uncertainty
Authors: Spiegelhalter D. J.
Published: July 2014

Sophisticated statistical insights are crucial for gaining knowledge from ever-larger data sets
Improving cardiac rhythm with a biological pacemaker
Authors: Munshi Nikhil V. , Olson Eric N.
Published: July 2014

Heart muscle cells in a large animal model are reprogrammed to restore heart rate and function
Topical Resiquimod Protects against Visceral Infection with Leishmania infantum chagasi in Mice
Authors: Craft Noah , Birnbaum Ron , Quanquin Natalie , Erfe Marie Crisel B. , Quant Cara , Haskell Jacquelyn , Bruhn Kevin W.
Published: July 2014

New prevention and treatment strategies are needed for visceral leishmaniasis, particularly ones that can be deployed simply and inexpensively in areas where leishmaniasis is endemic. Synthetic molecules that activate Toll-like receptor 7 and 8 (TLR7/8) pathways have previously been demonstrated to enhance protection against cutaneous leishmaniasis. We initially sought to determine whether the TLR7/8-activating molecule resiquimod might serve as an effective vaccine adjuvant targeting visceral leishmaniasis caused by infection withLeishmania infantum chagasi. Resiquimod was topically applied to the skin of mice either prior to or after systemic infection withL. infantum chagasi, and parasite burdens were assessed. Surprisingly, topical resiquimod application alone, in the absence of vaccination, conferred robust resistance to mice against future intravenous challenge with virulentL. infantum chagasi. This protection againstL. infantum chagasiinfection persisted as long as 8 weeks after the final topical resiquimod treatment. In addition, in mice with existing infections, therapeutic treatment with topical resiquimod led to significantly lower visceral parasite loads. Resiquimod increased trafficking of leukocytes, including B cells, CD4+and CD8+T cells, dendritic cells, macrophages, and granulocytes, in livers and spleens, which are the key target organs of visceralizing infection. We conclude that topical resiquimod leads to systemic immune modulation and confers durable protection against visceralizingL. infantum chagasiinfection, in both prophylactic and therapeutic settings. These studies support continued studies of TLR-modulating agents to determine mechanisms of protection and also provide a rationale for translational development of a critically needed, novel class of topical, preventative, and therapeutic agents for these lethal infections.
Glycosylated and Nonglycosylated Complement Control Protein of the Lister Strain of Vaccinia Virus
Authors: Meseda Clement A. , Kuhn Jordan , Atukorale Vajini , Campbell Joseph , Weir Jerry P.
Published: July 2014

The vaccinia virus complement control protein (VCP) is a secreted viral protein that binds the C3b and C4b complement components and inhibits the classic and alternative complement pathways. Previously, we reported that an attenuated smallpox vaccine, LC16m8, which was derived from the Lister strain of vaccinia virus (VV-Lister), expressed a glycosylated form of VCP, whereas published sequence data at that time indicated that the VV-Lister VCP has no motif for N-linked glycosylation. We were interested in determining whether the glycosylation of VCP impairs its biological activity, possibly contributing to the attenuation of LC16m8, and the likely origin of the glycosylated VCP. Expression analysis indicated that VV-Lister contains substrains expressing glycosylated VCP and substrains expressing nonglycosylated VCP. Other strains of smallpox vaccine, as well as laboratory strains of vaccinia virus, all expressed nonglycosylated VCP. Individual Lister virus clones expressing either the glycosylated VCP or the nonglycosylated species were isolated, and partially purified VCP from the isolates were found to be functional equivalents in binding human C3b and C4b complement proteins and inhibiting hemolysis and in immunogenicity. Recombinant vaccinia viruses expressing FLAG-tagged glycosylated VCP (FLAG-VCPg) and nonglycosylated VCP (FLAG-VCP) were constructed based on the Western Reserve strain. Purified FLAG-VCP and FLAG-VCPg bind human C3b and C4b and blocked complement-mediated hemolysis. Our data suggest that glycosylation did not affect the biological activity of VCP and thus may not have contributed to the attenuation of LC16m8. In addition, the LC16m8 virus likely originated from a substrain of VV-Lister that expresses glycosylated VCP.
The ID93 Tuberculosis Vaccine Candidate Does Not Induce Sensitivity to Purified Protein Derivative
Authors: Baldwin Susan L. , Reese Valerie , Granger Brian , Orr Mark T. , Ireton Gregory C. , Coler Rhea N. , Reed Steven G.
Published: July 2014

The tuberculin skin test (TST) is a simple and inexpensive test to determine whether individuals have been exposed toMycobacterium tuberculosis. This test is not always reliable, however, in people previously immunized with BCG and/or who have been exposed to environmental mycobacterial species due to a reaction to purified protein derivative (PPD) used in the skin test. An issue with BCG, therefore, is that the resulting sensitization to PPD in some individuals compromises the diagnostic use of the skin test. The ability to induce protective immune responses without sensitizing to the tuberculin skin test will be important properties of next-generation tuberculosis (TB) vaccine candidates. We show here that guinea pigs immunized with the candidate TB vaccine ID93/GLA-SE, currently in clinical trials, do not react to intradermal PPD administration. In contrast, positive DTH responses to both ID93 and components thereof were induced in ID93/GLA-SE-immunized animals, indicating robust but specific cellular responses were present in the immunized animals. Noninterference with the TST is an important factor for consideration in the development of a vaccine againstM. tuberculosis.
Use of the BacT/Alert System for Rapid Detection of Microbial Contamination in a Pilot Study Using Pancreatic Islet Cell Products
Authors: Murray Laura , McGowan Neil , Fleming John , Bailey Laura
Published: July 2014

At the Islet Isolation Laboratory of the Scottish National Blood Transfusion Service, manual sterility testing data show that contamination rates are 57.7% for pancreas transport fluid, 4.3% for postpurification islet samples, and 0% for pretransplant islet samples. This pilot study presents the BacT/Alert System as an alternative to manual testing to provide more rapid and sensitive sterility results for islet cell products.
Neonatal Infection with Neisseria meningitidis: Analysis of a 97-Year Period Plus Case Study
Authors: Kiray Baş Evrim , Bülbül Ali , Cömert Serdar , Uslu Sinan , Arslan Selda , Nuhoglu Asiye
Published: July 2014

Neisseria meningitidis
is one of the major causes of meningitis in children and adolescents, but it is rarely found during the neonatal period. Here, we describe a neonate with meningococcal sepsis who was admitted to the hospital on postnatal day 10, and we discuss the clinical features of neonatal infection with
N. meningitidis
in relation to the literature (analysis of a 97-year period).
Sequential Asymptomatic Enterovirus Infections in a Patient with Major Histocompatibility Complex Class II Primary Immunodeficiency
Authors: Driss Nadia , Mellouli Fethi , Ben Yahia Ahlem , Touzi Henda , Barbouche Mohamed-Ridha , Triki Henda , Bejaoui Mohamed
Published: July 2014

Patients with primary immunodeficiencies are usually susceptible to enterovirus infections and have higher risks to develop severe clinical forms. We report a unique description of a boy with major histocompatibility complex class II (MHC-II) deficiency infected by 9 different enterovirus serotypes during a 2-year period, with very mild clinical symptoms, probably due to the immunoglobulin therapy he was receiving.
Molecular Epidemiology of the Pertussis Epidemic in Washington State in 2012
Authors: Bowden Katherine E. , Williams Margaret M. , Cassiday Pamela K. , Milton Andrea , Pawloski Lucia , Harrison Marsenia , Martin Stacey W. , Meyer Sarah , Qin Xuan , DeBolt Chas , Tasslimi Azadeh , Syed Nusrat , Sorrell Ronald , Tran Mike , Hiatt Brian , Tondella Maria Lucia
Published: July 2014

Although pertussis disease is vaccine preventable, Washington State experienced a substantial rise in pertussis incidence beginning in 2011. By June 2012, the reported cases reached 2,520 (37.5 cases per 100,000 residents), a 1,300% increase compared with the same period in 2011. We assessed the molecular epidemiology of this statewide epidemic using 240 isolates collected from case patients reported from 19 of 39 Washington counties during 2012 to 2013. The typing methods included pulsed-field gel electrophoresis (PFGE), multilocus variable number tandem repeat analysis (MLVA), multilocus sequence typing (MLST), and pertactin gene (
prn
) mutational analysis. Using the scheme PFGE-MLVA-MLST-
prn
mutations-Prn deficiency, the 240 isolates comprised 65 distinct typing profiles. Thirty-one PFGE types were found, with the most common types, CDC013 (
n
= 51), CDC237 (
n
= 44), and CDC002 (
n
= 42), accounting for 57% of them. Eleven MLVA types were observed, mainly comprising type 27 (
n
= 183, 76%). Seven MLST types were identified, with the majority of the isolates typing as
prn2-ptxP3-ptxA1-fim3-1
(
n
= 157, 65%). Four different
prn
mutations accounted for the 76% of isolates exhibiting pertactin deficiency. PFGE provided the highest discriminatory power (
D
= 0.87) and was found to be a more powerful typing method than MLVA and MLST combined (
D
= 0.67). This study provides evidence for the continued predominance of MLVA 27 and
prn2-ptxP3-ptxA1
alleles, along with the reemergence of the
fim3-1
allele. Our results indicate that the
Bordetella pertussis
population causing this epidemic was diverse, with a few molecular types predominating. The PFGE, MLVA, and MLST profiles were consistent with the predominate types circulating in the United States and other countries. For
prn
, several mutations were present in multiple molecular types.
The Bristol Stool Scale and Its Relationship to Clostridium difficile Infection
Authors: Caroff Daniel A. , Edelstein Paul H. , Hamilton Keith , Pegues David A.
Published: July 2014

The Bristol stool form scale classifies the relative density of stool samples. In a prospective cohort study, we investigated the associations between stool density,
C. difficile
assay positivity, hospital-onset
C. difficile
infection, complications, and severity of
C. difficile
. We describe associations between the Bristol score, assay positivity, and clinical
C. difficile
infection.
Genetic and Molecular Predictors of High Vancomycin MIC in Staphylococcus aureus Bacteremia Isolates
Authors: Holmes Natasha E. , Turnidge John D. , Munckhof Wendy J. , Robinson J. Owen , Korman Tony M. , O'Sullivan Matthew V. N. , Anderson Tara L. , Roberts Sally A. , Warren Sanchia J. C. , Coombs Geoffrey W. , Tan Hui-Leen , Gao Wei , Johnson Paul D. R. , Howden Benjamin P.
Published: July 2014

An elevated vancomycin MIC is associated with poor outcomes in
Staphylococcus aureus
bacteremia (SAB) and is reported in patients with methicillin-susceptible
S. aureus
(MSSA) bacteremia in the absence of vancomycin treatment. Here, using DNA microarray and phenotype analysis, we investigated the genetic predictors and accessory gene regulator (
agr
) function and their relationship with elevated vancomycin MIC using blood culture isolates from a multicenter binational cohort of patients with SAB. Specific clonal complexes were associated with elevated (clonal complex 8 [CC8] [
P
< 0.001]) or low (CC22 [
P
< 0.001], CC88 [
P
< 0.001], and CC188 [
P
= 0.002]) vancomycin MIC.
agr
dysfunction (
P
= 0.014) or
agr
genotype II (
P
= 0.043) were also associated with an elevated vancomycin MIC. Specific resistance and virulence genes were also linked to an elevated vancomycin MIC, including
blaZ
(
P
= 0.002),
sea
(
P
< 0.001),
clfA
(
P
< 0.001),
splA
(
P
= 0.001), and the arginine catabolic mobile element (ACME) locus (
P
= 0.02). These data suggest that inherent organism characteristics may explain the link between elevated vancomycin MICs and poor outcomes in patients with SAB, regardless of the antibiotic treatment received. A consideration of clonal specificity should be included in future research when attempting to ascertain treatment effects or clinical outcomes.
Fast and Accurate Identification of Dermatophytes by Matrix-Assisted Laser Desorption Ionization–Time of Flight Mass Spectrometry: Validation in the Clinical Laboratory
Authors: Packeu A. , De Bel A. , l'Ollivier C. , Ranque S. , Detandt M. , Hendrickx M.
Published: July 2014

The performance of a matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF MS) workflow using an extensive reference database for dermatophyte identification was evaluated on 176 clinical strains. Using a direct-deposit procedure after 3 incubation days yielded 40% correct identification. Both increasing incubation time and using an extraction procedure resulted in 100% correct identification.
Up to Four Distinct Polypeptides Are Produced from the γ34.5 Open Reading Frame of Herpes Simplex Virus 2
Authors: Korom Maria , Davis Katie L. , Morrison Lynda A.
Published: July 2014

The herpes simplex virus 1 (HSV-1) ICP34.5 protein strongly influences neurovirulence and regulates several cellular antiviral responses. Despite the clinical importance of HSV-2, relatively little is known about its ICP34.5 ortholog. We found that HSV-2 produces up to four distinct forms of ICP34.5 in infected cells: a full-length protein, one shorter form sharing the N terminus, and two shorter forms sharing the C terminus. These forms appeared with similar kinetics and accumulated in cells over much of the replication cycle. We confirmed that the N-terminal form is translated from the primary unspliced transcript to a stop codon within the intron unique to HSV-2 γ34.5. We found that the N-terminal form was produced in a variety of cell types and by 9 of 10 clinical isolates. ICP27 influenced but was not required for expression of the N-terminal form. Western blotting and reverse transcription-PCR indicated the C-terminal forms did not contain the N terminus and were not products of alternative splicing or internal transcript initiation. Expression plasmids encoding methionine at amino acids 56 and 70 generated products that comigrated in SDS-PAGE with the C1 and C2 forms, respectively, and mutation of these sites abolished C1 and C2. Using a recombinant HSV-2 encoding hemagglutinin (HA)-tagged ICP34.5, we demonstrated that the C-terminal forms were also produced during infection of many human and mouse cell types but were not detectable in mouse primary neurons. The protein diversity generated from the HSV-2 γ34.5 open reading frame implies additional layers of cellular regulation through potential independent activities associated with the various forms of ICP34.5.

IMPORTANCE
The herpes simplex virus 1 (HSV-1) protein ICP34.5, encoded by the γ34.5 gene, interferes with several host defense mechanisms by binding cellular proteins that would otherwise stimulate the cell's autophagic, translational-arrest, and type I interferon responses to virus infection. ICP34.5 also plays a crucial role in determining the severity of nervous system infections with HSV-1 and HSV-2. The HSV-2 γ34.5 gene contains an intron not present in HSV-1 γ34.5. A shorter N-terminal form of HSV-2 ICP34.5 can be translated from the unspliced γ34.5 mRNA. Here, we show that two additional forms consisting of the C-terminal portion of ICP34.5 are generated in infected cells. Production of these N- and C-terminal forms is highly conserved among HSV-2 strains, including many clinical isolates, and they are broadly expressed in several cell types, but not mouse primary neurons. Multiple ICP34.5 polypeptides add additional complexity to potential functional interactions influencing HSV-2 neurovirulence.
High-Risk Human Papillomavirus E6 Protein Promotes Reprogramming of Fanconi Anemia Patient Cells through Repression of p53 but Does Not Allow for Sustained Growth of Induced Pluripotent Stem Cells
Authors: Chlon Timothy M. , Hoskins Elizabeth E. , Mayhew Christopher N. , Wikenheiser-Brokamp Kathryn A. , Davies Stella M. , Mehta Parinda , Myers Kasiani C. , Wells James M. , Wells Susanne I.
Published: July 2014

DNA repair plays a crucial role in embryonic and somatic stem cell biology and cell reprogramming. The Fanconi anemia (FA) pathway, which promotes error-free repair of DNA double-strand breaks, is required for somatic cell reprogramming to induced pluripotent stem cells (iPSC). Thus, cells from Fanconi anemia patients, which lack this critical pathway, fail to be reprogrammed to iPSC under standard conditions unless the defective FA gene is complemented. In this study, we utilized the oncogenes of high-risk human papillomavirus 16 (HPV16) to overcome the resistance of FA patient cells to reprogramming. We found that E6, but not E7, recovers FA iPSC colony formation and, furthermore, that p53 inhibition is necessary and sufficient for this activity. The iPSC colonies resulting from each of these approaches stained positive for alkaline phosphatase, NANOG, and Tra-1-60, indicating that they were fully reprogrammed into pluripotent cells. However, FA iPSC were incapable of outgrowth into stable iPSC lines regardless of p53 suppression, whereas their FA-complemented counterparts grew efficiently. Thus, we conclude that the FA pathway is required for the growth of iPSC beyond reprogramming and that p53-independent mechanisms are involved.

IMPORTANCE
A novel approach is described whereby HPV oncogenes are used as tools to uncover DNA repair-related molecular mechanisms affecting somatic cell reprogramming. The findings indicate that p53-dependent mechanisms block FA cells from reprogramming but also uncover a previously unrecognized defect in FA iPSC proliferation independent of p53.
Interferon Regulatory Factor 5-Dependent Immune Responses in the Draining Lymph Node Protect against West Nile Virus Infection
Authors: Thackray Larissa B. , Shrestha Bimmi , Richner Justin M. , Miner Jonathan J. , Pinto Amelia K. , Lazear Helen M. , Gale Michael , Diamond Michael S.
Published: July 2014

Upon activation of Toll-like and RIG-I-like receptor signaling pathways, the transcription factor IRF5 translocates to the nucleus and induces antiviral immune programs. The recent discovery of a homozygous mutation in the immunoregulatory gene guanine exchange factor dedicator of cytokinesis 2 (Dock2mu/mu) in severalIrf5−/−mouse colonies has complicated interpretation of immune functions previously ascribed to IRF5. To define the antiviral functions of IRF5in vivo, we infected backcrossedIrf5−/−×Dock2wt/wtmice (here calledIrf5−/−mice) and independently generated CMV-CreIrf5fl/flmice with West Nile virus (WNV), a pathogenic neurotropic flavivirus. Compared to congenic wild-type animals,Irf5−/−and CMV-CreIrf5fl/flmice were more vulnerable to WNV infection, and this phenotype was associated with increased infection in peripheral organs, which resulted in higher virus titers in the central nervous system. The loss of IRF5, however, was associated with only small differences in the type I interferon response systemically and in the draining lymph node during WNV infection. Instead, lower levels of several other proinflammatory cytokines and chemokines, as well as fewer and less activated immune cells, were detected in the draining lymph node 2 days after WNV infection. WNV-specific antibody responses inIrf5−/−mice also were blunted in the context of live or inactivated virus infection and this was associated with fewer antigen-specific memory B cells and long-lived plasma cells. Our results withIrf5−/−mice establish a key role for IRF5 in shaping the early innate immune response in the draining lymph node, which impacts the spread of virus infection, optimal B cell immunity, and disease pathogenesis.

IMPORTANCEAlthough the roles of IRF3 and IRF7 in orchestrating innate and adaptive immunity after viral infection are established, the function of the related transcription factor IRF5 remains less certain. Prior studies inIrf5−/−mice reported conflicting results as to the contribution of IRF5 in regulating type I interferon and adaptive immune responses. The lack of clarity may stem from a recently discovered homozygous loss-of-function mutation of the immunoregulatory geneDock2in several colonies ofIrf5−/−mice. Here, using a mouse model with a deficiency in IRF5 and wild-type Dock2 alleles, we investigated how IRF5 modulates West Nile virus (WNV) pathogenesis and host immune responses. Ourin vivostudies indicate that IRF5 has a key role in shaping the early proinflammatory cytokine response in the draining lymph node, which impacts immunity and control of WNV infection.
In Vivo
Reactivation of Latent Herpes Simplex Virus 1 in Mice Can Occur in the Brain before Occurring in the Trigeminal Ganglion
Authors: Yao Hui-Wen , Ling Pin , Tung Yuk-Ying , Hsu Sheng-Min , Chen Shun-Hua
Published: July 2014

Herpes simplex virus 1 (HSV-1) establishes latency in neurons of the brains and sensory ganglia of humans and experimentally infected mice. The latent virus can reactivate to cause recurrent infection. Both primary and recurrent infections can induce diseases, such as encephalitis. In humans, the majority of encephalitis cases occur as a recurrent infection. However, in the past, numerous mouse studies documented that viral reactivation occurs efficiently in the ganglion, but extremely rarely in the brain, when assessed
ex vivo
by cultivating minced tissue explants. Here, we compare the brains and the trigeminal ganglia of mice latently infected with HSV-1 (strain 294.1 or McKrae) for levels of viral genomes and
in vivo
reactivation. The numbers of copies of 294.1 and McKrae genomes in the brain stem were significantly greater than those in the trigeminal ganglion. Most importantly, 294.1 and McKrae reactivation was detected in the brain stems earlier than in the trigeminal ganglia of mice treated with hyperthermia to reactivate latent virus
in vivo
. In addition, the brain stem yielded reactivated virus at a high frequency compared with the trigeminal ganglion, especially in mice latently infected with 294.1 after hyperthermia treatment. These results provide evidence that recurrent brain infection can be induced by the reactivation of latent virus in the brain
in situ
.

IMPORTANCE
Herpes simplex virus 1 (HSV-1) establishes latency in neurons of the brains and sensory ganglia of humans and experimentally infected mice. The latent virus can reactivate to cause recurrent infection. In the past, studies of viral reactivation focused on the ganglion, because efficient viral reactivation was detected in the ganglion but not in the brain when assessed
ex vivo
by cultivating mouse tissue explants. In this study, we report that the brain contains more viral genomes than the trigeminal ganglion in latently infected mice. Notably, the brain yields reactivated virus early and efficiently compared with the trigeminal ganglion after mice are stimulated to reactivate latent virus. Our findings raise the potential importance of HSV-1 latent infection and reactivation in the brain.
Shielding of a Lipooligosaccharide IgM Epitope Allows Evasion of Neutrophil-Mediated Killing of an Invasive Strain of Nontypeable Haemophilus influenzae
Authors: Langereis Jeroen D. , Weiser Jeffrey N.
Published: July 2014

Nontypeable
Haemophilus influenzae
is a frequent cause of noninvasive mucosal inflammatory diseases but may also cause invasive diseases, such as sepsis and meningitis, especially in children and the elderly. Infection by nontypeable
Haemophilus influenzae
is characterized by recruitment of neutrophilic granulocytes. Despite the presence of a large number of neutrophils, infections with nontypeable
Haemophilus influenzae
are often not cleared effectively by the antimicrobial activity of these immune cells. Herein, we examined how nontypeable
Haemophilus influenzae
evades neutrophil-mediated killing. Transposon sequencing (Tn-seq) was used on an isolate resistant to neutrophil-mediated killing to identify genes required for its survival in the presence of human neutrophils and serum, which provided a source of complement and antibodies. Results show that nontypeable
Haemophilus influenzae
prevents complement-dependent neutrophil-mediated killing by expression of surface galactose-containing oligosaccharide structures. These outer-core structures block recognition of an inner-core lipooligosaccharide epitope containing glucose attached to heptose HepIII-β1,2-Glc by replacement with galactose attached to HepIII or through shielding HepIII-β1,2-Glc by phase-variable attachment of oligosaccharide chain extensions. When the HepIII-β1,2-Glc-containing epitope is expressed and exposed, nontypeable
Haemophilus influenzae
is opsonized by naturally acquired IgM generally present in human serum and subsequently phagocytosed and killed by human neutrophils. Clinical nontypeable
Haemophilus influenzae
isolates containing galactose attached to HepIII that are not recognized by this IgM are more often found to cause invasive infections.

IMPORTANCE
Neutrophils are white blood cells that specialize in killing pathogens and are recruited to sites of inflammation. However, despite the presence of large numbers of neutrophils in the middle ear cavity and lungs of patients with otitis media or chronic obstructive pulmonary disease, respectively, the bacterium nontypeable
Haemophilus influenzae
is often not effectively cleared from these locations by these immune cells. In order to understand how nontypeable
Haemophilus influenzae
is able to cause inflammatory diseases in the presence of neutrophils, we determined the mechanism that underlies resistance to neutrophil-mediated killing. We have shown that nontypeable
Haemophilus influenzae
prevents binding of antibodies of the IgM subtype through changes in their surface lipooligosaccharide structure, thereby preventing complement activation and clearance by human neutrophils.
Parallel Evolution of Streptococcus pneumoniae and Streptococcus mitis to Pathogenic and Mutualistic Lifestyles
Authors: Kilian Mogens , Riley David R. , Jensen Anders , Brüggemann Holger , Tettelin Hervé
Published: July 2014

The bacterium
Streptococcus pneumoniae
is one of the leading causes of fatal infections affecting humans. Intriguingly, phylogenetic analysis shows that the species constitutes one evolutionary lineage in a cluster of the otherwise commensal
Streptococcus mitis
strains, with which humans live in harmony. In a comparative analysis of 35 genomes, including phylogenetic analyses of all predicted genes, we have shown that the pathogenic pneumococcus has evolved into a master of genomic flexibility while lineages that evolved into the nonpathogenic
S. mitis
secured harmonious coexistence with their host by stabilizing an approximately 15%-reduced genome devoid of many virulence genes. Our data further provide evidence that interspecies gene transfer between
S. pneumoniae
and
S. mitis
occurs in a unidirectional manner, i.e., from
S. mitis
to
S. pneumoniae
. Import of genes from
S. mitis
and other mitis, anginosus, and salivarius group streptococci ensured allelic replacements and antigenic diversification and has been driving the evolution of the remarkable structural diversity of capsular polysaccharides of
S. pneumoniae
. Our study explains how the unique structural diversity of the pneumococcal capsule emerged and conceivably will continue to increase and reveals a striking example of the fragile border between the commensal and pathogenic lifestyles. While genomic plasticity enabling quick adaptation to environmental stress is a necessity for the pathogenic streptococci, the commensal lifestyle benefits from stability.

IMPORTANCE
One of the leading causes of fatal infections affecting humans,
Streptococcus pneumoniae
, and the commensal
Streptococcus mitis
are closely related obligate symbionts associated with hominids. Faced with a shortage of accessible hosts, the two opposing lifestyles evolved in parallel. We have shown that the nonpathogenic
S. mitis
secured harmonious coexistence with its host by stabilizing a reduced genome devoid of many virulence genes. Meanwhile, the pathogenic pneumococcus evolved into a master of genomic flexibility and imports genes from
S. mitis
and other related streptococci. This process ensured antigenic diversification and has been driving the evolution of the remarkable structural diversity of capsular polysaccharides of
S. pneumoniae
, which conceivably will continue to increase and present a challenge to disease prevention.
Association of socioeconomic status measured by education, and cardiovascular health: a population-based cross-sectional study
Authors: Janković Slavenka , Stojisavljević Dragana , Janković Janko , Erić Miloš , Marinković Jelena
Published: July 2014

Objective

Cardiovascular health (CVH) is a relatively new concept defined by the American Heart Association (AHA). The aim of the present study was to assess whether the indices of CVH were discriminators of socioeconomic status (SES) in the adult population of the Republic of Srpska (RS).

Design

Population-based cross-sectional study.

Setting

RS, Bosnia and Herzegovina.

Participants

The study involved 4165 adults aged ≥18 years (mean age 50.2; 54% women) who participated in the National Health Survey performed from September to November 2010 in the RS.

Study variables

Participant's education was a proxy for SES. Potential discriminators of SES were indices of CVH presented according to AHA as: ideal health behaviours index (non-smoking, body mass index <25 kg/m2, physical activity at goal level and healthy diet); ideal health factors index (untreated total cholesterol <200 mg/dL, untreated blood pressure <120/<80 mm Hg, untreated fasting glucose <100 mg/dL and non-smoking); and ideal CVH status (defined as all seven ideal health metrics present) versus intermediate and poor CVH status.

Results

Participants with high educational levels had a significantly greater number of ideal CVH metrics, and ideal health factor metrics compared with those with low or medium educational level (OR 0.88 95% CI 0.77 to 0.99 and OR 0.88 95% CI 0.80 to 0.96; OR 0.81 95% CI 0.69 to 0.96 and OR 0.77 95% CI 0.68 to 0.87; respectively). The number of ideal behaviour metrics was not a discriminator of educational groups. Concerning the categories of CVH status the poor CVH was a discriminator for low and medium education compared with those with high education (OR 1.93 95% CI 1.24 to 3.01 and OR 1.54 95% CI 1.08 to 2.19, respectively).

Conclusions

Our findings emphasise the large potential for preventing cardiovascular disease, showing a low proportion with a favourable CVH profile, especially among low-educated people. It is necessary to consider prevention strategies aimed at improving CVH in RS, targeting primarily low educational groups.
The effect of a behaviour change intervention on the diets and physical activity levels of women attending Sure Start Children’s Centres: results from a complex public health intervention
Authors: Baird Janis , Jarman Megan , Lawrence Wendy , Black Christina , Davies Jenny , Tinati Tannaze , Begum Rufia , Mortimore Andrew , Robinson Sian , Margetts Barrie , Cooper Cyrus , Barker Mary , Inskip Hazel
Published: July 2014

Objectives

The UK government's response to the obesity epidemic calls for action in communities to improve people's health behaviour. This study evaluated the effects of a community intervention on dietary quality and levels of physical activity of women from disadvantaged backgrounds.

Design

Non-randomised controlled evaluation of a complex public health intervention.

Participants

527 women attending Sure Start Children's Centres (SSCC) in Southampton (intervention) and 495 women attending SSCCs in Gosport and Havant (control).

Intervention

Training SSCC staff in behaviour change skills that would empower women to change their health behaviours.

Outcomes

Main outcomesdietary quality and physical activity.Intermediate outcomesself-efficacy and sense of control.

Results

1-year post-training, intervention staff used skills to support behaviour change significantly more than control staff. There were statistically significant reductions of 0.1 SD in the dietary quality of all women between baseline and follow-up and reductions in self-efficacy and sense of control. The decline in self-efficacy and control was significantly smaller in women in the intervention group than in women in the control group (adjusted differences in self-efficacy and control, respectively, 0.26 (95% CI 0.001 to 0.50) and 0.35 (0.05 to 0.65)). A lower decline in control was associated with higher levels of exposure in women in the intervention group. There was a statistically significant improvement in physical activity in the intervention group, with 22.9% of women reporting the highest level of physical activity compared with 12.4% at baseline, and a smaller improvement in the control group. The difference in change in physical activity level between the groups was not statistically significant (adjusted difference 1.02 (0.74 to 1.41)).

Conclusions

While the intervention did not improve women's diets and physical activity levels, it had a protective effect on intermediate factors—control and self-efficacy—suggesting that a more prolonged exposure to the intervention might improve health behaviour. Further evaluation in a more controlled setting is justified.
Is there a causal link between knee loading and knee osteoarthritis progression? A systematic review and meta-analysis of cohort studies and randomised trials
Authors: Henriksen Marius , Creaby Mark W , Lund Hans , Juhl Carsten , Christensen Robin
Published: July 2014

Objective

We performed a systematic review, meta-analysis and assessed the evidence supporting a causal link between knee joint loading during walking and structural knee osteoarthritis (OA) progression.

Design

Systematic review, meta-analysis and application of Bradford Hill's considerations on causation.

Data sources

We searched MEDLINE, Scopus, AMED, CINAHL and SportsDiscus for prospective cohort studies and randomised controlled trials (RCTs) from 1950 through October 2013.

Study eligibility criteria

We selected cohort studies and RCTs in which estimates of knee joint loading during walking were used to predict structural knee OA progression assessed by X-ray or MRI.

Data analyses

Meta-analysis was performed to estimate the combined OR for structural disease progression with higher baseline loading. The likelihood of a causal link between knee joint loading and OA progression was assessed from cohort studies using the Bradford Hill guidelines to derive a 0–4 causation score based on four criteria and examined for confirmation in RCTs.

Results

Of the 1078 potentially eligible articles, 5 prospective cohort studies were included. The studies included a total of 452 patients relating joint loading to disease progression over 12–72 months. There were very serious limitations associated with the methodological quality of the included studies. The combined OR for disease progression was 1.90 (95% CI 0.85 to 4.25; I2=77%) for each one-unit increment in baseline knee loading. The combined causation score was 0, indicating no causal association between knee loading and knee OA progression. No RCTs were found to confirm or refute the findings from the cohort studies.

Conclusions

There is very limited and low-quality evidence to support for a causal link between knee joint loading during walking and structural progression of knee OA.

Trial registration number

CRD42012003253