bluuebunny/crossref_metadata_2025_split huggingface dataset:
Synovial fluid chondroitin sulphate indicates abnormal joint metabolism in asymptomatic osteochondritic horses
Authors: MACHADO T. S. L. , CORREIA da SILVA L. C. L. , BACCARIN R. Y. A. , MICHELACCI Y. M. 
Published: January 2012

SummaryReasons for performing study:Alternative methods to evaluate the joint condition in asymptomatic osteochondrosis dissecans (OCD) and other joint diseases may be useful.Objectives:To investigate possible changes in synovial fluid composition that may lead to joint conditions in asymptomatic OCD, in mature horses.Methods:Animals aged >2 years, of different breeds, with OCD in the intermediate ridge of distal tibia, symptomatic or not, were studied. Synovial fluid samples (10 healthy; 11 asymptomatic OCD; 25 symptomatic OCD) were collected by arthroscopy from 29 horses. Glycosaminoglycans (GAGs) were analysed by a combination of agarose gel electrophoresis and enzymatic degradation with specific GAG lyases. The viscosity, white blood cell (WBC) count, protein concentration and hyaluronic acid (HA) molecular weight were also determined.Results:The method used here to analyse synovial fluid GAGs is reliable, reproducible and specific. The main synovial fluid GAGs are HA and chondroitin sulphate (CS), 93% and 7% respectively in normal horses. In symptomatic OCD, the concentrations of both increased (expressed as GAG/urea ratios), but CS increased more. The CS increased also in asymptomatic OCD. An inflammatory reaction was suggested by the increased WBC counts in OCD. The molecular weight of the synovial fluid HA was reduced in OCD, explaining the lower viscosity observed.Conclusions:The increased CS in synovial fluid of OCD joints in mature horses suggests that the synovial fluid CS and the WBC count are good markers of the joint conditions, allowing the identification of pathological phase in joint diseases.Potential relevance:The analysis of synovial fluid GAGs shows that cartilage damage occurs even in asymptomatic OCD, implying that arthroscopic removal of osteochondral fragments should be performed even in asymptomatic OCD.
A recess intervention to promote moderate‐to‐vigorous physical activity
Authors: Howe C. A. , Freedson P. S. , Alhassan S. , Feldman H. A. , Osganian S. K. 
Published: January 2012

SummaryBackgroundSchools provide a prime environment for interventions that attempt to increase physical activity and prevent obesity.ObjectiveThis study examined the effect of a 30‐min, structured recess using 22 games of known energy expenditure on moderate‐to‐vigorous physical activity when compared to free play implemented with third graders from two elementary schools over 9 weeks.MethodsModerate‐to‐vigorous physical activity and other cardiovascular risk factors were assessed pre‐ and post‐intervention in 27 children.ResultsModerate‐to‐vigorous physical activity during recess increased significantly in intervention school children from 6.9 ± 0.8 to 14.9 ± 0.9 min pre‐ and post‐intervention, respectively (adjusted mean change 8.0 ± 1.1; P < 0.0001), with no differences by gender or body mass index (BMI). In‐school, moderate‐to‐vigorous physical activity also increased significantly more for intervention compared to control children (adjusted mean change 14 ± 4 min vs. 3 ± 3 min; P = 0.014, respectively).ConclusionThere was no significant difference in BMI and cardiovascular risk factors. A structured recess is feasible to implement and can significantly increase moderate‐to‐vigorous physical activity.
Phenotypic Screening of Carbapenemases and Associated β-Lactamases in Carbapenem-Resistant Enterobacteriaceae
Authors: Birgy André , Bidet Philippe , Genel Nathalie , Doit Catherine , Decré Dominique , Arlet Guillaume , Bingen Edouard 
Published: January 2012

ABSTRACT

Dissemination of carbapenem resistance among
Enterobacteriaceae
poses a considerable threat to public health. Carbapenemase gene detection by molecular methods is the gold standard but is available in only a few laboratories. The aim of this study was to test phenotypic methods for the detection of metallo-β-lactamase (MBL)- or
Klebsiella pneumoniae
carbapenemase (KPC)-producing
Enterobacteriaceae
and associated mechanisms of β-lactam resistance against a panel of 30 genotypically characterized carbapenem-resistant
Enterobacteriaceae
: 9 MBL, 7 KPC, 6 OXA-48, and 8 extended-spectrum β-lactamase (ESBL) or AmpC β-lactamases associated with decreased permeability. We used carbapenemase inhibitor-impregnated agar to test for carbapenem-resistant strains. Differences in the inhibition zone sizes of the meropenem, imipenem, ertapenem, and doripenem disks were measured between control and inhibitor (EDTA or phenylboronic acid [PBA] with or without cloxacillin)-impregnated Mueller-Hinton agar with a cutoff of 10 mm. All 9 MBL- and 7 KPC-producing
Enterobacteriaceae
were identified from the differences in zone size in the presence and absence of specific inhibitors, regardless of the carbapenem MICs and including isolates with low-level resistance to carbapenems. We also detected their associated β-lactam resistance mechanisms (11 ESBL-type and 5 class A β-lactamase 2b). No differences in zone size were observed for OXA-48-producing strains or other carbapenem resistance mechanisms such as ESBL and decreased permeability. We propose a new strategy to detect carbapenemases (MBL- and KPC-type) and associated mechanisms of β-lactam resistance (ESBL or class A β-lactamase 2b) by the use of inhibitor-impregnated agar. A rapid phenotypic detection of resistance mechanisms is important for epidemiological purposes and for limiting the spread of resistant strains by implementing specific infection control measures.
Phenotypic, Biochemical, and Molecular Techniques for Detection of Metallo-β-Lactamase NDM in Acinetobacter baumannii
Authors: Bonnin Rémy A. , Naas Thierry , Poirel Laurent , Nordmann Patrice 
Published: January 2012

ABSTRACT

Seven NDM-positive
Acinetobacter baumannii
isolates of worldwide origin were studied to evaluate the best technique for their identification. Detection of carbapenemase producers based on the measurement of carbapenemase activity by UV spectrophotometry (as for
A. baumannii
strains producing other types of carbapenemase), or by the modified Hodge test, failed. Inhibition activity using EDTA was a sensitive technique but lacked specificity compared to molecular techniques, which remain the gold standard.
Evidence for N-Glycan Shielding of Antigenic Sites during Evolution of Human Influenza A Virus Hemagglutinin
Authors: Kobayashi Yuki , Suzuki Yoshiyuki 
Published: January 2012

ABSTRACT

After the emergence of influenza A viruses in the human population, the number of N-glycosylation sites (NGS) in the globular head region of hemagglutinin (HA) has increased continuously for several decades. It has been speculated that the addition of NGS to the globular head region of HA has conferred selective advantages to the virus by preventing the binding of antibodies (Ab) to antigenic sites (AS). Here, the effect of N-glycosylation on the binding of Ab to AS in human influenza A virus subtype H3N2 (A/H3N2) was examined by inferring natural selection at AS and other sites (NAS) that are located close to and distantly from the NGS in the three-dimensional structure of HA through a comparison of the rates of synonymous (
d
S
) and nonsynonymous (
d
N
) substitutions. When positions 63, 122, 126, 133, 144, and 246 in the globular head region of HA were non-NGS, the
d
N
/
d
S
was >1 and positive selection was detected at the AS located near these positions. However, the
d
N
/
d
S
value decreased and the evidence of positive selection disappeared when these positions became NGS. In contrast,
d
N
/
d
S
at the AS distantly located from the positions mentioned above and at the NAS of any location were generally <1 and did not decrease when these positions changed from non-NGS to NGS. These results suggest that the attachment of N-glycans to the NGS in the globular head region of HA prevented the binding of Ab to AS in the evolutionary history of human A/H3N2 virus.
HIV Cell-to-Cell Transmission Requires the Production of Infectious Virus Particles and Does Not Proceed through Env-Mediated Fusion Pores
Authors: Monel Blandine , Beaumont Elodie , Vendrame Daniela , Schwartz Olivier , Brand Denys , Mammano Fabrizio 
Published: January 2012

ABSTRACT
Direct cell-to-cell transmission of human immunodeficiency virus (HIV) is a more potent and efficient means of virus propagation than infection by cell-free virus particles. The aim of this study was to determine whether cell-to-cell transmission requires the assembly of enveloped virus particles or whether nucleic acids with replication potential could translocate directly from donor to target cells through envelope glycoprotein (Env)-induced fusion pores. To this end, we characterized the transmission properties of viruses carrying mutations in the matrix protein (MA) that affect the incorporation of Env into virus particles but do not interfere with Env-mediated cell-cell fusion. By use of cell-free virus, the infectivity of MA mutant viruses was below the detection threshold both in single-cycle and in multiple-cycle assays. Truncation of the cytoplasmic tail (CT) of Env restored the incorporation of Env into MA mutant viruses and rescued their cell-free infectivity to different extents. In cell-to-cell transmission assays, MA mutations prevented HIV transmission from donor to target cells, despite efficient Env-dependent membrane fusion. HIV transmission was blocked at the level of virus core translocation into the cytosol of target cells. As in cell-free assays, rescue of Env incorporation by truncation of the Env CT restored the virus core translocation and cell-to-cell infectivity of MA mutant viruses. These data show that HIV cell-to-cell transmission requires the assembly of enveloped virus particles. The increased efficiency of this infection route may thus be attributed to the high local concentrations of virus particles at sites of cellular contacts rather than to a qualitatively different transmission process.
Role for the Phosphatidylinositol 3-Kinase-Akt-TOR Pathway during Sindbis Virus Replication in Arthropods
Authors: Patel Rohini K. , Hardy Richard W. 
Published: January 2012

ABSTRACT

The efficient transmission of alphaviruses requires the establishment of a persistent infection in the arthropod vector; however, the nature of the virus-arthropod host interaction is not well understood. The phosphatidylinositol 3-kinase (PI3K)-Akt-TOR pathway is a signaling pathway with which viruses interact to manipulate cellular functions. The viral activation of this pathway can enhance translation and inhibit apoptosis, potentially promoting viral replication; conversely, repression can enhance cell death. Using a system to study Sindbis virus RNA replication in
Drosophila melanogaster
, we found that the overexpression of Akt enhanced Sindbis virus replication. In contrast, a decrease in viral replication was observed for flies hypomorphic for the Akt gene. Infection of cultured
Drosophila
cells led to the phosphorylation and activation of Akt. The chemical inhibition of PI3K, Akt, and TOR in mosquito cells reduced virus replication, suggesting that this pathway is proviral. Early after infection, there was an increase in the TOR-dependent phosphorylation of 4E-BP1 in mosquito cells and a consequent increase in the translation of a capped reporter mRNA. In contrast, no change in 4E-BP1 phosphorylation was seen in mammalian cells, and the level of translation of the reporter decreased following infection. Finally, we found that the increase in the phosphorylation of 4E-BP1 was stimulated by replicon RNA but not by UV-inactivated virus. Our data indicate that Sindbis virus replication complex formation in mosquito cells activates the PI3K-Akt-TOR pathway, causing the phosphorylation of 4E-BP1 and increasing the formation of eukaryotic initiation factor 4F (eIF4F), which promote cap-dependent translation. This virus-induced increase in cap-dependent translation allows the efficient translation of viral mRNA while minimizing the burden on the cell.
Enterovirus 71 Disrupts Interferon Signaling by Reducing the Level of Interferon Receptor 1
Authors: Lu Jing , Yi Lina , Zhao Jin , Yu Jun , Chen Ying , Lin Marie C. , Kung Hsiang-Fu , He Ming-Liang 
Published: January 2012

ABSTRACTThe recent outbreak of enterovirus 71 (EV71) infected millions of children and caused over 1,000 deaths. To date, neither an effective vaccine nor antiviral treatment is available for EV71 infection. Interferons (IFNs) have been successfully applied to treat patients with hepatitis B and C viral infections for decades but have failed to treat EV71 infections. Here, we provide the evidence that EV71 antagonizes type I IFN signaling by reducing the level of interferon receptor 1 (IFNAR1). We show that the host cells could sense EV71 infection and stimulate IFN-β production. However, the induction of downstream IFN-stimulated genes is inhibited by EV71. Also, only a slight interferon response and antiviral effects could be detected in cells treated with recombinant type I IFNs after EV71 infection. Further studies reveal that EV71 blocks the IFN-mediated phosphorylation of STAT1, STAT2, Jak1, and Tyk2 by reducing IFNAR1. Finally, we identified the 2A protease encoded by EV71 as an antagonist of IFNs and show that the protease activity is required for reducing IFNAR1 levels. Taken together, our study for the first time uncovers a mechanism used by EV71 to antagonize type I IFN signaling and provides new targets for future antiviral strategies.
The 3A Protein from Multiple Picornaviruses Utilizes the Golgi Adaptor Protein ACBD3 To Recruit PI4KIIIβ
Authors: Greninger Alexander L. , Knudsen Giselle M. , Betegon Miguel , Burlingame Alma L. , DeRisi Joseph L. 
Published: January 2012

ABSTRACT
The activity of phosphatidylinositol 4-kinase class III beta (PI4KIIIβ) has been shown to be required for the replication of multiple picornaviruses; however, it is unclear whether a physical association between PI4KIIIβ and the viral replication machinery exists and, if it does, whether association is necessary. We examined the ability of the 3A protein from 18 different picornaviruses to form a complex with PI4KIIIβ by affinity purification of Strep-Tagged transiently transfected constructs followed by mass spectrometry and Western blotting for putative interacting targets. We found that the 3A proteins of Aichi virus, bovine kobuvirus, poliovirus, coxsackievirus B3, and human rhinovirus 14 all copurify with PI4KIIIβ. Furthermore, we found that multiple picornavirus 3A proteins copurify with the Golgi adaptor protein acyl coenzyme A (acyl-CoA) binding domain protein 3 (ACBD3/GPC60), including those from Aichi virus, bovine kobuvirus, human rhinovirus 14, poliovirus, and coxsackievirus B2, B3, and B5. Affinity purification of ACBD3 confirmed interaction with multiple picornaviral 3A proteins and revealed the ability to bind PI4KIIIβ in the absence of 3A. Mass-spectrometric analysis of transiently expressed Aichi virus, bovine kobuvirus, and human klassevirus 3A proteins demonstrated that the N-terminal glycines of these 3A proteins are myristoylated. Alanine-scanning mutagenesis along the entire length of Aichi virus 3A followed by transient expression and affinity purification revealed that copurification of PI4KIIIβ could be eliminated by mutation of specific residues, with little or no effect on recruitment of ACBD3. One mutation at the N terminus, I5A, significantly reduced copurification of both ACBD3 and PI4KIIIβ. The dependence of Aichi virus replication on the activity of PI4KIIIβ was confirmed by both chemical and genetic inhibition. Knockdown of ACBD3 by small interfering RNA (siRNA) also prevented replication of both Aichi virus and poliovirus. Point mutations in 3A that eliminate PI4KIIIβ association sensitized Aichi virus to PIK93, suggesting that disruption of the 3A/ACBD3/PI4KIIIβ complex may represent a novel target for therapeutic intervention that would be complementary to the inhibition of the kinase activity itself.
Expression of Dicer and Drosha in triple-negative breast cancer
Authors: Passon Nadia , Gerometta Anna , Puppin Cinzia , Lavarone Elisa , Puglisi Fabio , Tell Gianluca , Di Loreto Carla , Damante Giuseppe 
Published: January 2012

AimsDicer and Drosha are components of the miRNA-producing machinery and their altered expression may play a role in cancer progression. The main purpose of this study was a detailed investigation of Dicer and Drosha expression and localisation in triple-negative breast cancers.MethodsThirty-one triple-negative breast cancers and several breast cancer cell lines were investigated. Expression of Dicer and Drosha was evaluated at the mRNA level by quantitative reverse transcription PCR and at the protein level by immunohistochemistry or western blot.ResultsCompared with normal breast tissues, a wide variation of Dicer and Drosha mRNA levels was detected in triple-negative breast cancers. As a group, Drosha mRNA levels in triple-negative breast cancers were significantly higher than those in normal breast tissues. Immunohistochemical data confirmed higher expression of Drosha protein in triple-negative breast cancers. In normal breast tissues Dicer was detectable predominantly in the cytoplasm of basal/myoepithelial cells only. In contrast, in the majority of triple-negative breast cancers, intense Dicer staining was detectable also in the nuclear compartment. Detection of Dicer and Drosha mRNA and protein levels in breast cancer cell lines confirmed the nuclear localisation of Dicer, suggesting, in addition, that the steady-state protein levels could be controlled by post-mRNA regulatory events.ConclusionsThese findings indicate that Dicer and Drosha expression is deregulated in triple-negative breast cancers.
Optical properties of anatase TiO2 films modified by N ion implantation
Authors: Xiang X. , Chang D. , Jiang Y. , Liu C.M. , Zu X.T. 
Published: January 2012

Anatase TiO2 thin films are deposited on K9 glass samples at different substrate temperatures by radio frequency magnetron sputtering. N ion implantation is performed in the as-deposited TiO2 thin films at ion fluences of 5 × 1016, 1 × 1017, and 5 × 1017 ions/cm2. X-ray diffraction, atomic force microscope, X-ray photoelectron spectroscopy (XPS), and UV–visible spectrophotometer are used to characterize the films. With increasing N ion fluences, the absorption edges of anatase TiO2 films shift to longer wavelengths and the absorbance increases in the visible light region. XPS results show that the red shift of TiO2 films is due to the formation of N–Ti–O compounds. As a result, photoactivity is enhanced with increasing N ion fluence.
Analysis of temperature-dependent Schottky barrier parameters of Cu–Au Schottky contacts to n-InP
Authors: Devi V. Lakshmi , Jyothi I. , Reddy V. Rajagopal 
Published: January 2012

In this work, we have investigated the electrical characteristics of Au–Cu–n-InP Schottky contacts by current–voltage (I–V) and capacitance–voltage (C–V) measurements in the temperature range 260–420 K in steps of 20 K. The diode parameters, such as the ideality factor, n, and zero-bias barrier height, Φb0, have been found to be strongly temperature dependent. It has been found that the zero-bias barrier height, Φb0(I–V), increases and the ideality factor, n, decreases with an increase in temperature. The forward I–V characteristics are analyzed on the basis of standard thermionic emission (TE) theory and the assumption of gaussian distribution of barrier heights, due to barrier inhomogeneities that prevail at the metal–semiconductor interface. The zero-bias barrier height Φb0 versus 1/2kT plot has been drawn to obtain the evidence of a gaussian distribution of the barrier heights. The corresponding values are Φb0 = 1.16 eV and σ0 = 159 meV for the mean barrier height and standard deviation, respectively. The modified Richardson plot has given mean barrier height, Φb0, and Richardson constant, A**, as 1.15 eV and 7.34 Acm−2K−2, respectively, which is close to the theoretical value of 9.4 Acm−2K−2. Barrier heights obtained from C–V measurements are higher than those obtained from I–V measurements. This inconsistency between Schottky barrier heights (SBHs) obtained from I–V and C–V measurements was also interpreted. The temperature dependence of the I–V characteristics of the Au–Cu–n-InP Schottky diode has been explained on the basis of TE mechanism with gaussian distribution of the SBHs.
Targeting Pioneering Factor and Hormone Receptor Cooperative Pathways to Suppress Tumor Progression
Authors: Shah Supriya , Prasad Shikha , Knudsen Karen E. 
Published: January 2012

Abstract
Nuclear receptors and pioneer factors drive the development and progression of prostate cancer. In this disease, aggressive disease phenotypes and hormone therapy failures result from resurgent activity of androgen receptor (AR) and the upregulation of coactivator protein p300 and pioneer factors (e.g., GATA2 and FOXA1). Thus, a major emphasis in the field is to identify mechanisms by which castrate-resistant AR activity and pioneer factor function can be combinatorially suppressed. Here we show that the turmeric spice isoflavone curcumin suppresses p300 and CBP occupancy at sites of AR function. Curcumin reduced the association of histone acetylation and pioneer factors, thereby suppressing AR residence and downstream target gene expression. Histone deacetylase inhibitors reversed the effects of curcumin on AR activity, further underscoring the impact of curcumin on altering the chromatin landscape. These functions precluded pioneer factor occupancy, leading ultimately to a suppression of ligand-dependent and ligand-independent AR residence on chromatin. Moreover, these functions were conserved even in cells with heightened pioneer factor activity, thus identifying a potential strategy to manage this subclass of tumors. Biological relevance was further identified using in vivo xenograft models mimicking disease progression. Curcumin cooperated in vivo with androgen deprivation as indicated by a reduction in tumor growth and delay to the onset of castrate-resistant disease. Together, our results show the combinatorial impact of targeting AR and histone modification in prostate cancer, thus setting the stage for further development of curcumin as a novel agent to target AR signaling. Cancer Res; 72(5); 1248–59. ©2012 AACR.
ATR–ATRIP Kinase Complex Triggers Activation of the Fanconi Anemia DNA Repair Pathway
Authors: Shigechi Tomoko , Tomida Junya , Sato Koichi , Kobayashi Masahiko , Eykelenboom John K. , Pessina Fabio , Zhang Yanbin , Uchida Emi , Ishiai Masamichi , Lowndes Noel F. , Yamamoto Kenichi , Kurumizaka Hitoshi , Maehara Yoshihiko , Takata Minoru 
Published: January 2012

Abstract
ATR kinase activates the S-phase checkpoint when replication forks stall at sites of DNA damage. This event also causes phosphorylation of the Fanconi anemia (FA) protein FANCI, triggering its monoubiquitination of the key DNA repair factor FANCD2 by the FA core E3 ligase complex, thereby promoting this central pathway of DNA repair which permits replication to be restarted. However, the interplay between ATR and the FA pathway has been unclear. In this study, we present evidence that their action is directly linked, gaining insights into this relationship in a DT40 mutant cell line that is conditionally deficient in the critical ATR-binding partner protein ATRIP. Using this system, we showed that ATRIP was crucial for DNA damage–induced FANCD2 monoubiquitination and FANCI phosphorylation. ATR kinase phosphorylated recombinant FANCI protein in vitro, which was facilitated by the presence of FANCD2. Mechanistic investigations revealed that the RPA region but not the TopBP1 region of ATRIP was required for FANCD2 monoubiquitination, whereas Chk1 phosphorylation relied upon both domains. Together, our findings identify ATR as the kinase responsible for activating the FA pathway of DNA repair. Cancer Res; 72(5); 1149–56. ©2012 AACR.