bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pannexin 1 channels mediate the release of ATP into the lumen of the rat urinary bladder
Authors: Beckel Jonathan M. , Daugherty Stephanie L. , Tyagi Pradeep , Wolf‐Johnston Amanda S. , Birder Lori A. , Mitchell Claire H. , de Groat William C. 
Published: January 2015

Key points
ATP is released through pannexin channels into the lumen of the rat urinary bladder in response to distension or stimulation with bacterial endotoxins.
Luminal ATP plays a physiological role in the control of micturition because intravesical perfusion of apyrase or the ecto‐ATPase inhibitor ARL67156 altered reflex bladder activity in the anaesthetized rat.
The release of ATP from the apical and basolateral surfaces of the urothelium appears to be mediated by separate mechanisms because intravesical administration of the pannexin channel antagonist Brilliant Blue FCF increased bladder capacity, whereas i.v. administration did not.
Intravesical instillation of small interfering RNA‐containing liposomes decreased pannexin 1 expression in the rat urothelium in vivo and increased bladder capacity.
These data indicate a role for pannexin‐mediated luminal ATP release in both the physiological and pathophysiological control of micturition and suggest that urothelial pannexin may be a viable target for the treatment of overactive bladder disorders.
AbstractATP is released from the bladder epithelium, also termed the urothelium, in response to mechanical or chemical stimuli. Although numerous studies have described the contribution of this release to the development of various bladder disorders, little information exists regarding the mechanisms of release. In the present study, we examined the role of pannexin channels in mechanically‐induced ATP release from the urothelium. PCR confirmed the presence of pannexin 1 and 2 mRNA in rat urothelial tissue, whereas immunofluorescence experiments localized pannexin 1 to all three layers of the urothelium. During continuous bladder cystometry in anaesthetized rats, inhibition of pannexin 1 channels using carbenoxolone (CBX) or Brilliant Blue FCF (BB‐FCF) (1–100 μm, intravesically), or by using intravesical small interfering RNA, increased the interval between voiding contractions. Intravenous administration of BB‐FCF (1–100 μg kg−1) did not alter bladder activity. CBX or BB‐FCF (100 μm intravesically) also decreased basal ATP concentrations in the perfusate from non‐distended bladders and inhibited increases in ATP concentrations in response to bladder distension (15 and 30 cmH2O pressure). Intravesical perfusion of the ATP diphosphohydrolase apyrase (2 U ml−1), or the ATPase inhibitor ARL67156 (10 μm) increased or decreased reflex bladder activity, respectively. Intravesical instillation of bacterial lipopolysaccharides (LPS) (Escherichia coli 055:B5, 100 μg ml−1) increased ATP concentrations in the bladder perfusate, and also increased voiding frequency; these effects were suppressed by BB‐FCF. These data indicate that pannexin channels contribute to distension‐ or LPS‐evoked ATP release into the lumen of the bladder and that luminal release can modulate voiding function.
Increased muscle blood supply and transendothelial nutrient and insulin transport induced by food intake and exercise: effect of obesity and ageing
Authors: Wagenmakers Anton J. M. , Strauss Juliette A. , Shepherd Sam O. , Keske Michelle A. , Cocks Matthew 
Published: January 2015

AbstractThis review concludes that a sedentary lifestyle, obesity and ageing impair the vasodilator response of the muscle microvasculature to insulin, exercise and VEGF‐A and reduce microvascular density. Both impairments contribute to the development of insulin resistance, obesity and chronic age‐related diseases. A physically active lifestyle keeps both the vasodilator response and microvascular density high. Intravital microscopy has shown that microvascular units (MVUs) are the smallest functional elements to adjust blood flow in response to physiological signals and metabolic demands on muscle fibres. The luminal diameter of a common terminal arteriole (TA) controls blood flow through up to 20 capillaries belonging to a single MVU. Increases in plasma insulin and exercise/muscle contraction lead to recruitment of additional MVUs. Insulin also increases arteriolar vasomotion. Both mechanisms increase the endothelial surface area and therefore transendothelial transport of glucose, fatty acids (FAs) and insulin by specific transporters, present in high concentrations in the capillary endothelium. Future studies should quantify transporter concentration differences between healthy and at risk populations as they may limit nutrient supply and oxidation in muscle and impair glucose and lipid homeostasis. An important recent discovery is that VEGF‐B produced by skeletal muscle controls the expression of FA transporter proteins in the capillary endothelium and thus links endothelial FA uptake to the oxidative capacity of skeletal muscle, potentially preventing lipotoxic FA accumulation, the dominant cause of insulin resistance in muscle fibres.
Suitability of poroelastic and viscoelastic mechanical models for high and low frequency MR elastography
Authors: McGarry M. D. J. , Johnson C. L. , Sutton B. P. , Georgiadis J. G. , Van Houten E. E. W. , Pattison A. J. , Weaver J. B. , Paulsen K. D. 
Published: January 2015

Purpose:Descriptions of the structure of brain tissue as a porous cellular matrix support application of a poroelastic (PE) mechanical model which includes both solid and fluid phases. However, the majority of brain magnetic resonance elastography (MRE) studies use a single phase viscoelastic (VE) model to describe brain tissue behavior, in part due to availability of relatively simple direct inversion strategies for mechanical property estimation. A notable exception is low frequency intrinsic actuation MRE, where PE mechanical properties are imaged with a nonlinear inversion algorithm.Methods:This paper investigates the effect of model choice at each end of the spectrum of in vivo human brain actuation frequencies. Repeat MRE examinations of the brains of healthy volunteers were used to compare image quality and repeatability for each inversion model for both 50 Hz externally produced motion and ≈1 Hz intrinsic motions. Additionally, realistic simulated MRE data were generated with both VE and PE finite element solvers to investigate the effect of inappropriate model choice for ideal VE and PE materials.Results:In vivo, MRE data revealed that VE inversions appear more representative of anatomical structure and quantitatively repeatable for 50 Hz induced motions, whereas PE inversion produces better results at 1 Hz. Reasonable VE approximations of PE materials can be derived by equating the equivalent wave velocities for the two models, provided that the timescale of fluid equilibration is not similar to the period of actuation. An approximation of the equilibration time for human brain reveals that this condition is violated at 1 Hz but not at 50 Hz. Additionally, simulation experiments when using the “wrong” model for the inversion demonstrated reasonable shear modulus reconstructions at 50 Hz, whereas cross‐model inversions at 1 Hz were poor quality. Attenuation parameters were sensitive to changes in the forward model at both frequencies, however, no spatial information was recovered because the mechanisms of VE and PE attenuation are different.Conclusions:VE inversions are simpler with fewer unknown properties and may be sufficient to capture the mechanical behavior of PE brain tissue at higher actuation frequencies. However, accurate modeling of the fluid phase is required to produce useful mechanical property images at the lower frequencies of intrinsic brain motions.
In vivo demonstration of magnetic guidewire steerability in a MRI system with additional gradient coils
Authors: Lalande Viviane , Gosselin Frederick P. , Vonthron Manuel , Conan Benjamin , Tremblay Charles , Beaudoin Gilles , Soulez Gilles , Martel Sylvain 
Published: January 2015

Purpose:To assess the ability to control the steering of a modified guidewire actuated by the magnetic force of a magnetic resonance imaging system with additional gradient coils for selective arterial catheterization in rabbits.Methods:Selective catheterizations of the right renal artery, left renal artery, superior mesenteric artery, and iliac artery were performed on two rabbits. A 3D magnetic force was applied onto a magnetic bead placed at the tip of a guidewire. The ability of the guidewire to advance in the aorta without entering the side branches when the magnetic force was not applied was also evaluated. Steering of the guidewire was combined with a dedicated tracking system and its position was registered on the 3D model of a magnetic resonance angiography (MRA).Results:The magnetic catheterization of the renal arteries was successful and showed reproducibility. Superior mesenteric artery and iliac artery showed that the catheterization was feasible. These two arteries were difficult to visualize on MRA, making catheterization and setting the direction of the force more difficult. There was no inadvertent catheterization of side vessels when the guidewire was advanced with magnetic steering despite the hook shape at the tip of the guidewire caused by the alignment of the bead anisotropy with the permanent magnetic field.Conclusions:This first evaluation of selective catheterization of aortic branches with a magnetic guidewire provided a successful steering in the less angled side branches and this modified guidewire was advanced in the aorta without inadvertent selective catheterization when manipulated without magnetic actuation.
Protective Immunogenicity of Group A Streptococcal M-Related Proteins
Authors: Dale James B. , Niedermeyer Shannon E. , Agbaosi Tina , Hysmith Nicholas D. , Penfound Thomas A. , Hohn Claudia M. , Pullen Matthew , Bright Michael I. , Murrell Daniel S. , Shenep Lori E. , Courtney Harry S. 
Published: January 2015

ABSTRACTMany previous studies have focused on the surface M proteins of group A streptococci (GAS) as virulence determinants and protective antigens. However, the majority of GAS isolates express M-related protein (Mrp) in addition to M protein, and both have been shown to be required for optimal virulence. In the current study, we evaluated the protective immunogenicity of Mrp to determine its potential as a vaccine component that may broaden the coverage of M protein-based vaccines. Sequence analyses of 33mrpgenes indicated that there are three families of structurally related Mrps (MrpI, MrpII, and MrpIII). N-terminal peptides of Mrps were cloned, expressed, and purified from M type 2 (M2) (MrpI), M4 (MrpII), and M49 (MrpIII) GAS. Rabbit antisera against the Mrps reacted at high titers with the homologous Mrp, as determined by enzyme-linked immunosorbent assay, and promoted bactericidal activity against GASemmtypes expressing Mrps within the same family. Mice passively immunized with rabbit antisera against MrpII were protected against challenge infections with M28 GAS. Assays for Mrp antibodies in serum samples from 281 pediatric subjects aged 2 to 16 indicated that the Mrp immune response correlated with increasing age of the subjects. Affinity-purified human Mrp antibodies promoted bactericidal activity against a number of GAS representing differentemmtypes that expressed an Mrp within the same family but showed no activity againstemmtypes expressing an Mrp from a different family. Our results indicate that Mrps have semiconserved N-terminal sequences that contain bactericidal epitopes which are immunogenic in humans. These findings may have direct implications for the development of GAS vaccines.
Mycobacterial DNA Extraction for Whole-Genome Sequencing from Early Positive Liquid (MGIT) Cultures
Authors: Votintseva Antonina A. , Pankhurst Louise J. , Anson Luke W. , Morgan Marcus R. , Gascoyne-Binzi Deborah , Walker Timothy M. , Quan T. Phuong , Wyllie David H. , Del Ojo Elias Carlos , Wilcox Mark , Walker A. Sarah , Peto Tim E. A. , Crook Derrick W. 
Published: January 2015

ABSTRACT

We developed a low-cost and reliable method of DNA extraction from as little as 1 ml of early positive mycobacterial growth indicator tube (MGIT) cultures that is suitable for whole-genome sequencing to identify mycobacterial species and predict antibiotic resistance in clinical samples. The DNA extraction method is based on ethanol precipitation supplemented by pretreatment steps with a MolYsis kit or saline wash for the removal of human DNA and a final DNA cleanup step with solid-phase reversible immobilization beads. The protocol yielded ≥0.2 ng/μl of DNA for 90% (MolYsis kit) and 83% (saline wash) of positive MGIT cultures. A total of 144 (94%) of the 154 samples sequenced on the MiSeq platform (Illumina) achieved the target of 1 million reads, with <5% of reads derived from human or nasopharyngeal flora for 88% and 91% of samples, respectively. A total of 59 (98%) of 60 samples that were identified by the national mycobacterial reference laboratory (NMRL) as
Mycobacterium tuberculosis
were successfully mapped to the H37Rv reference, with >90% coverage achieved. The DNA extraction protocol, therefore, will facilitate fast and accurate identification of mycobacterial species and resistance using a range of bioinformatics tools.
Reevaluation of Commercial Reagents for Detection of Histoplasma capsulatum Antigen in Urine
Authors: Theel Elitza S. , Harring Julie A. , Dababneh Ala S. , Rollins Leonard O. , Bestrom Jeannie E. , Jespersen Deborah J. 
Published: January 2015

ABSTRACT

Detection of the
Histoplasma capsulatum
urinary antigen (UAg) is among the most sensitive and rapid means to diagnose histoplasmosis. Previously, we evaluated analyte-specific reagents (ASR) manufactured by IMMY (Norman, OK) for detection of
Histoplasma
galactomannan (GM) in urine using an enzyme immunoassay (EIA), and we showed low positive agreement (64.5%) with the MiraVista (MVista)
Histoplasma
antigen (Ag) quantitative EIA (MiraVista Diagnostics, Indianapolis, IN). Here we reevaluated the IMMY GM ASR following modification of our original assay protocol and introduction of an indeterminate range. A total of 150 prospectively collected urine samples were tested with both the IMMY and MVista EIAs, and clinical histories were recorded for all study subjects. The IMMY GM ASR showed positive and negative agreements of 82.3% (14/17 samples) and 100% (121/121 samples), respectively (with exclusion of 12 indeterminate results), and overall agreement of 90% (135/150 samples) with respect to the MVista EIA. Of the three patients with negative IMMY GM ASR results and positive MVista EIA results, testing was performed for initial diagnostic purposes for one patient (<0.4 ng/ml by the MVista EIA) and UAg levels were being monitored for the remaining two patients (both <0.7 ng/ml by the MVista EIA). The MVista EIA results were positive for 6/12 samples that tested indeterminate by the IMMY GM ASR. We also show that the IMMY GM ASR can be used to serially monitor
Histoplasma
UAg levels. In conclusion, we demonstrate that, with modification, the IMMY GM ASR is a reliable rapid assay for detection of
Histoplasma
UAg.
Tracing Melioidosis Back to the Source: Using Whole-Genome Sequencing To Investigate an Outbreak Originating from a Contaminated Domestic Water Supply
Authors: McRobb Evan , Sarovich Derek S. , Price Erin P. , Kaestli Mirjam , Mayo Mark , Keim Paul , Currie Bart J. 
Published: January 2015

ABSTRACT

Melioidosis, a disease of public health importance in Southeast Asia and northern Australia, is caused by the Gram-negative soil bacillus
Burkholderia pseudomallei
. Melioidosis is typically acquired through environmental exposure, and case clusters are rare, even in regions where the disease is endemic.
B. pseudomallei
is classed as a tier 1 select agent by the Centers for Disease Control and Prevention; from a biodefense perspective, source attribution is vital in an outbreak scenario to rule out a deliberate release. Two cases of melioidosis within a 3-month period at a residence in rural northern Australia prompted an investigation to determine the source of exposure.
B. pseudomallei
isolates from the property's groundwater supply matched the multilocus sequence type of the clinical isolates. Whole-genome sequencing confirmed the water supply as the probable source of infection in both cases, with the clinical isolates differing from the likely infecting environmental strain by just one single nucleotide polymorphism (SNP) each. For the first time, we report a phylogenetic analysis of genomewide insertion/deletion (indel) data, an approach conventionally viewed as problematic due to high mutation rates and homoplasy. Our whole-genome indel analysis was concordant with the SNP phylogeny, and these two combined data sets provided greater resolution and a better fit with our epidemiological chronology of events. Collectively, this investigation represents a highly accurate account of source attribution in a melioidosis outbreak and gives further insight into a frequently overlooked reservoir of
B. pseudomallei
. Our methods and findings have important implications for outbreak source tracing of this bacterium and other highly recombinogenic pathogens.
Evaluation of Portability and Cost of a Fluorescent PCR Ribotyping Protocol for Clostridium difficile Epidemiology
Authors: Martinson Jonathan N. V. , Broadaway Susan , Lohman Egan , Johnson Christina , Alam M. Jahangir , Khaleduzzaman Mohammed , Garey Kevin W. , Schlackman Jessica , Young Vincent B. , Santhosh Kavitha , Rao Krishna , Lyons Robert H. , Walk Seth T. 
Published: January 2015

ABSTRACT

Clostridium difficile
is the most commonly identified pathogen among health care-associated infections in the United States. There is a need for accurate and low-cost typing tools that produce comparable data across studies (i.e., portable data) to help characterize isolates during epidemiologic investigations of
C. difficile
outbreaks and sporadic cases of disease. The most popular
C. difficile
-typing technique is PCR ribotyping, and we previously developed methods using fluorescent PCR primers and amplicon sizing on a Sanger-style sequencer to generate fluorescent PCR ribotyping data. This technique has been used to characterize tens of thousands of
C. difficile
isolates from cases of disease. Here, we present validation of a protocol for the cost-effective generation of fluorescent PCR ribotyping data. A key component of this protocol is the ability to accurately identify PCR ribotypes against an online database (
http://walklab.rcg.montana.edu
) at no cost. We present results from a blinded multicenter study to address data portability across four different laboratories and three different sequencing centers. Our standardized protocol and centralized database for typing of
C. difficile
pathogens will increase comparability between studies so that important epidemiologic linkages between cases of disease and patterns of emergence can be rapidly identified.
Disruption of Type I Interferon Signaling by the Nonstructural Protein of Severe Fever with Thrombocytopenia Syndrome Virus via the Hijacking of STAT2 and STAT1 into Inclusion Bodies
Authors: Ning Yun-Jia , Feng Kuan , Min Yuan-Qin , Cao Wu-Chun , Wang Manli , Deng Fei , Hu Zhihong , Wang Hualin 
Published: January 2015

ABSTRACT
The type I interferon (IFN) system, including IFN induction and signaling, is the critical component of the host defense line against viral infection, which, in turn, is also a vulnerable target for viral immune evasion. Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging bunyavirus. Previous data have shown that SFTSV can interfere with the early induction of type I IFNs through targeting host kinases TBK1/IKKε. In this study, we demonstrated that SFTSV also can suppress type I IFN-triggered signaling and interferon-stimulated gene (ISG) expression. Interestingly, we observed the significant inhibition of IFN signaling in cells transfected with the plasmids encoding the nonstructural protein (NSs) but not the nucleocapsid protein (NP), indicating the role of NSs as an antagonist of IFN signaling. Furthermore, coimmunoprecipitation (Co-IP) and pulldown assays indicated that NSs interacts with the cellular signal transducer and activator of transcription 2 (STAT2), and the DNA-binding domain of STAT2 may contribute to the NSs-STAT2 interaction. Combined with confocal microscopy analyses, we demonstrated that NSs sequesters STAT2 and STAT1 into viral inclusion bodies (IBs) and impairs IFN-induced STAT2 phosphorylation and nuclear translocation of both STATs, resulting in the inhibition of IFN signaling and ISG expression. SFTSV NSs-mediated hijacking of STATs in IBs represents a novel mechanism of viral suppression of IFN signaling, highlighting the role of viral IBs as the virus-built “jail” sequestering some crucial host factors and interfering with the corresponding cellular processes.

IMPORTANCE
SFTSV is an emerging bunyavirus which can cause a severe hemorrhagic fever-like disease with high case fatality rates in humans, posing a serious health threat. However, there are no specific antivirals available, and the pathogenesis and virus-host interactions are largely unclear. Here, we demonstrated that SFTSV can inhibit type I IFN antiviral signaling by the NSs-mediated hijacking of STAT2 and STAT1 into viral IBs, highlighting the interesting role of viral IBs in virus-host interactions as the virus-built jail. Sequestering signaling molecules into IBs represents a novel and, perhaps, also a general mechanism of viral suppression of IFN signaling, the understanding of which may benefit the study of viral pathogenesis and the development of antiviral therapies.