bluuebunny/crossref_metadata_2025_split huggingface dataset:
PGE2 Contributes to In vitro MSC‐Mediated Inhibition of Non‐Specific and Antigen‐Specific T Cell Proliferation in MS Patients
Authors: Zafranskaya M. , Nizheharodava D. , Yurkevich M. , Ivanchik G. , Demidchik Y. , Kozhukh H. , Fedulov A. 
Published: August 2013

AbstractCurrent theories of multiple sclerosis (MS) induction and progression place autoreactive T cells in the focus of the pathogenesis. Mesenchymal/stromal stem cells (MSC) have become a promising alternative approach for pathogenic therapy of MS due to their immunomodulatory properties, underlying mechanisms of which are intensive study. The objective of the research was to investigate the contribution of PGE2 to MSC‐mediated suppression in patients with MS using in vitro model of mitogen‐ and myelin‐stimulated T cell cocultivation with autologous/allogeneic MSC. We have showed that PGE2 production depends on cell‐to‐cell contact of MSC and lymphocytes. The antigenic stimulation did not affect PGE2 production following cocultivation of MSC and PBMC, and it is the presence of MSC in cell culture that significantly increases PGE2 production irrespective of antigenic cultivation conditions. Simultaneously, PGE2 synthesis correlated with indexes of MSC‐mediated suppression of mitogen‐ and myelin‐stimulated T cell proliferation in patients with MS. No significant differences in PGE2 production by autologous and allogeneic MSC have been established. These results have demonstrated that in patients with MS, PGE2 is one of the possible factors of MSC immunosuppression. The interrelation between PGE2 concentrations and T cell proliferation suppression mediated by MSC may explain one of the immune mechanisms of cell therapy, which is crucial for the further proper use of MSC in MS research and pathogenic treatment.
Endogenous fibrinolytic potential in tissue‐plasminogen activator‐modified thromboelastography analysis is significantly decreased in dogs suffering from diseases predisposing to thrombosis
Authors: Spodsberg Eva H. , Wiinberg Bo , Jessen Lisbeth R. , Marschner Clara B. , Kristensen Annemarie T. 
Published: August 2013

BackgroundIn people, studies have shown that resistance to fibrinolysis could be a contributing factor to thrombosis. Tissue‐plasminogen‐activated (t‐PA) thromboelastography (TEG) has been used to evaluate endogenous fibrinolytic potential. In dogs, TEG has been used for the diagnosis of various hemostatic disorders, but studies evaluating fibrinolysis are limited. Investigations into the potential of t‐PA‐modified TEG to monitor endogenous fibrinolytic potential are lacking in both healthy dogs and dogs with diseases predisposing to development of thrombosis.ObjectivesThe aim of this study was to compare 3 t‐PA‐modified TEG assays and compare the endogenous fibrinolytic potential in dogs suffering from diseases associated with thrombosis with a group of healthy dogs.MethodsThree different TEG assays, such as native, tissue factor‐activated, and kaolin‐activated, were modified with t‐PA and used to compare whole blood samples from 16 healthy control dogs and 20 diseased dogs.ResultsThromboelastography lysis variables were significantly affected by addition of t‐PA in all 3 assays. Lysis results in diseased dogs were comparable to those in healthy dogs prior to addition of t‐PA. After addition of t‐PA, lysis results were significantly decreased in the diseased group compared with healthy dogs. The lowest median lysis levels were found in dogs with systemic inflammation and protein‐losing disorders.ConclusionAddition of t‐PA activates fibrinolysis in TEG of blood from both healthy dogs and dogs with diseases predisposing to thrombosis. The significantly decreased fibrinolysis in diseased dogs suggests that this may be a potential prothrombotic risk factor in dogs.
Manufacturing porcine islets: culture at 22 °C has no advantage above culture at 37 °C: a gene expression evaluation
Authors: Mueller Kate R. , Martins Kyra V. , Murtaugh Michael P. , Schuurman Henk‐Jan , Papas Klearchos K. 
Published: August 2013

AbstractBackgroundThe manufacturing process of islets includes a culture step which was originally introduced to ease the logistics of procedures in preparing the graft and transplant recipient. It has been suggested that culture at room temperature has an advantage over culture at 37 °C, in part by reducing immunogenicity via preferential elimination of contaminating cells (such as passenger leukocytes) within islets. We investigated this using islets isolated from pancreata of adult pigs.MethodsPorcine islets were isolated from three donors and cultured at 37 °C for 1 day, and then under three different conditions: 37 °C for 6 days (condition A); 22 °C for 6 days (condition B); or 22 °C for 5 days followed by 37 °C for 1 day (condition C). Recovery was assessed by DNA measurement, viability by oxygen consumption rate normalized for DNA (OCR/DNA), and gene expression by RT–PCR for a series of 9 lymphocyte markers, 11 lymphokines and chemokines, and 14 apoptotic and stress markers.ResultsPost‐culture islet recoveries were similar for the three culture conditions. Average OCR/DNA values were 129–159 nmol/min·mgDNA before culture, and 259–291, 204–212, and 207–228 nmol/min·mgDNA, respectively, for culture under conditions A, B, and C, respectively. Irrespective of culture condition, examined gene expression in all three series of lymphocyte markers, lymphokines and chemokines, and apoptotic and stress markers manifested a statistically significant decrease upon culture for 7 days. This decrease was most dramatic for condition A: in particular, most of lymphocyte markers showed a >10‐fold reduction and also six markers in the lymphokine and chemokine series; these reductions are consistent with the elimination of immune cells present within islets during culture. The reduction was less for apoptotic and stress markers. For culture under condition B, the reduction in gene expression was less, and culture under condition C resulted in gene expression levels similar to those under condition A: this indicates that 24 h at 37 °C is sufficient to re‐equilibrate gene expression levels from those in islets cultured at 22 °C to those in islets cultured at 37 °C. Results were consistent among the preparations from the three donors.ConclusionsCulture of porcine islets at 37 °C provides benefits over culture at 22 °C with respect to OCR/DNA outcomes and reduced expression of genes encoding lymphocyte markers, lymphokines and chemokines, and markers for apoptosis and stress.
Characterization and Optimization of the Glucan Particle-Based Vaccine Platform
Authors: Huang Haibin , Ostroff Gary R. , Lee Chrono K. , Specht Charles A. , Levitz Stuart M. 
Published: August 2013

ABSTRACTGlucan particles (GPs) are hollow porousSaccharomyces cerevisiaecell walls that are treated so that they are composed primarily of β-1,3-d-glucans. Our previous studies showed that GPs can serve as an effective vaccine platform. Here, we characterize CD4+T-cell and antibody responses in immunized mice as a function of antigen (ovalbumin) encapsulation, antigen dose, particle numbers, time, immunization schedule, and trapping methods. Although we found that GPs served as an effective adjuvant when admixed with free antigens for IgG1 antibody production, stronger CD4+T-cell and IgG2c antibody responses were stimulated when antigens were encapsulated inside GPs, suggesting that the GP platform acts as both an adjuvant and a delivery system. Vigorous T-cell and antibody responses were stimulated even at submicrogram antigen doses, as long as the number of GPs was kept at 5 × 107particles per immunization. One prime and one boost were sufficient to elicit robust immune responses. In addition, strong antigen-specific antibody and T-cell responses prevailed up to 20 months following the last immunization, including those of gamma interferon (IFN-γ), interleukin 17A (IL-17A), and dual IFN-γ/IL-17A-secreting CD4+T cells. Finally, robust immune responses were observed using generally recognized as safe (GRAS) materials (alginate and calcium, with or without chitosan) to trap antigens within GPs. Thus, these studies demonstrate that antigens encapsulated into GPs make an effective vaccine platform that combines adjuvanticity and antigen delivery to elicit strong durable immune responses at relatively low antigen doses using translationally relevant formulations.
In Vitro
Activity of Amikacin against Isolates of Mycobacterium avium Complex with Proposed MIC Breakpoints and Finding of a 16S rRNA Gene Mutation in Treated Isolates
Authors: Brown-Elliott Barbara A. , Iakhiaeva Elena , Griffith David E. , Woods Gail L. , Stout Jason E. , Wolfe Cameron R. , Turenne Christine Y. , Wallace Richard J. 
Published: August 2013

ABSTRACT

Amikacin is a major drug used for the treatment of
Mycobacterium avium
complex (MAC) disease, but standard laboratory guidelines for susceptibility testing are not available. This study presents
in vitro
amikacin MICs for 462 consecutive clinical isolates of the MAC using a broth microdilution assay. Approximately 50% of isolates had amikacin MICs of 8 μg/ml, and 86% had MICs of ≤16 μg/ml. Of the eight isolates (1.7%) with MICs of 64 μg/ml, five had an MIC of 32 μg/ml on repeat testing. Ten isolates (2.1%) had an initial amikacin MIC of >64 μg/ml, of which seven (1.5%) had MICs of >64 μg/ml on repeat testing. These seven isolates had a 16S rRNA gene A1408G mutation and included
M. avium
,
Mycobacterium intracellulare
, and
Mycobacterium chimaera
. Clinical data were available for five of these seven isolates, all of which had received prolonged (>6 months) prior therapy, with four that were known to be treated with amikacin. The 16S mutation was not detected in isolates with MICs of ≤64 μg/ml. We recommend primary testing of amikacin against isolates of the MAC and propose MIC guidelines for breakpoints that are identical to the CLSI guidelines for
Mycobacterium abscessus
: ≤16 μg/ml for susceptible, 32 μg/ml for intermediate, and ≥64 μg/ml for resistant. If considered and approved by the CLSI, this will be only the second drug recommended for primary susceptibility testing against the MAC and should facilitate its use for both intravenous and inhaled drug therapies.
Assessment of New Culture Method for Detection of Borrelia Species from Serum of Lyme Disease Patients
Authors: Johnson Barbara J. B. , Pilgard Mark A. , Russell Theresa M. 
Published: August 2013

ABSTRACT

A novel method of culturing spirochetes from the serum of U.S. Lyme disease patients was recently reported by Sapi and colleagues to have 94% sensitivity and 100% specificity for
Borrelia
species as assessed by microscopy and DNA sequence analysis of the
pyrG
gene (E. Sapi, N. Pabbati, A. Datar, E. M. Davies, A. Rattelle, and B. A. Kuo, Int. J. Med. Sci. 10:362–376, 2013). The majority of the spirochetes described were related by
pyrG
sequences to species of
Borrelia
previously undetected in North American patients without a reported history of travel to Europe or Asia. To better understand these unexpected findings, we determined
pyrG
sequences of the laboratory reference strains used by the investigators for method development and testing of culture medium. Eighty percent (41/51) of the reported patient-derived
pyrG
sequences were identical to one of the laboratory strains, and an additional 12% (6/51) differed by only a single nucleotide across a 603-bp region of the
pyrG
gene. Thus, false positivity due to laboratory contamination of patient samples cannot be ruled out, and further validation of the proposed novel culture method is required.
In SituEvolution of Virus-Specific Cytotoxic T Cell Responses in the Lung
Authors: Frey Stefanie , Pircher Hanspeter , Follo Marie , Collins Peter , Krempl Christine , Ehl Stephan 
Published: August 2013

ABSTRACTCytotoxic T cells (CTL) play a critical role in the clearance of respiratory viral infections, but they also contribute to disease manifestations. In this study, we infected mice with a genetically modified pneumonia virus of mice (PVM) that allowed visualization of virus-specific CTL and infected cellsin situ. The first virus-specific T cells entered the lung via blood vessels in the scattered foci of PVM-infected cells, which densely clustered around the bronchi at day 7 after infection. At this time, overall pulmonary virus load was maximal, but the mice showed no overt signs of disease. On days 8 to 9, T cells gained access to the infected bronchial epithelium and to the lung interstitium, which was associated with a reduction in the number of virus-infected cells within the initial clusters but could not prevent further virus spread throughout the lung tissue. Interestingly, recruitment of virus-specific CTL throughout the parenchyma was still ongoing on day 10, when the virus infection was already largely controlled. This also represented the peak of clinical disease. Thus, disease was associated with an exuberant T cell infiltration late in the course of the infection, which may be required to completely eliminate virus at residual foci of infection. PVM-induced immunopathology may thus result from the need to generate widespread T cell infiltrates to complete the elimination of virus-infected cells in a large organ like the lung. This experimental model provides the first insights into the spatiotemporal evolution of pulmonary antiviral T cell immunityin vivo.
Soluble Interleukin-6 Receptor-Mediated Innate Immune Response to DNA and RNA Viruses
Authors: Wang Qing , Chen Xueyuan , Feng Jian , Cao Yanhua , Song Yu , Wang Hui , Zhu Chengliang , Liu Shi , Zhu Ying 
Published: August 2013

ABSTRACTThe interleukin-6 (IL-6) receptor, which exists as membrane-bound and soluble forms, plays critical roles in the immune response. The soluble IL-6 receptor (sIL6R) has been identified as a potential therapeutic target for preventing coronary heart disease. However, little is known about the role of this receptor during viral infection. In this study, we show that sIL6R, but not IL-6, is induced by viral infection via the cyclooxygenase-2 pathway. Interestingly, sIL6R, but not IL-6, exhibited extensive antiviral activity against DNA and RNA viruses, including hepatitis B virus, influenza virus, human enterovirus 71, and vesicular stomatitis virus. No synergistic effects on antiviral action were observed by combining sIL6R and IL-6. Furthermore, sIL6R mediated antiviral action via the p28 pathway and induced alpha interferon (IFN-α) by promoting the nuclear translocation of IFN regulatory factor 3 (IRF3) and NF-κB, which led to the activation of downstream IFN effectors, including 2′,5′-oligoadenylate synthetase (OAS), double-stranded RNA-dependent protein kinase (PKR), and myxovirus resistance protein (Mx). Thus, our results demonstrate that sIL6R, but not IL-6, plays an important role in the host antiviral response.
Structural and Functional Studies on the Interaction of Adenovirus Fiber Knobs and Desmoglein 2
Authors: Wang Hongjie , Yumul Roma , Cao Hua , Ran Liang , Fan Xiaolong , Richter Maximilian , Epstein Forrest , Gralow Julie , Zubieta Chloe , Fender Pascal , Lieber André 
Published: August 2013

ABSTRACT

Human adenovirus (Ad) serotypes Ad3, Ad7, Ad11, and Ad14, as well as a recently emerged strain of Ad14 (Ad14p1), use the epithelial junction protein desmoglein 2 (DSG2) as a receptor for infection. Unlike Ad interaction with CAR and CD46, structural details for Ad binding to DSG2 are still elusive. Using an approach based on
Escherichia coli
expression libraries of random Ad3 and Ad14p1 fiber knob mutants, we identified amino acid residues that, when mutated individually, ablated or reduced Ad knob binding to DSG2. These residues formed three clusters inside one groove at the extreme distal end of the fiber knob. The Ad3 fiber knob mutant library was also used to identify variants with increased affinity to DSG2. We found a number of mutations within or near the EF loop of the Ad3 knob that resulted in affinities to DSG2 that were several orders of magnitude higher than those to the wild-type Ad3 knob. Crystal structure analysis of one of the mutants showed that the introduced mutations make the EF loop more flexible, which might facilitate the interaction with DSG2. Our findings have practical relevance for cancer therapy. We have recently reported that an Ad3 fiber knob-containing recombinant protein (JO-1) is able to trigger opening of junctions between epithelial cancer cells which, in turn, greatly improved the intratumoral penetration and efficacy of therapeutic agents (I. Beyer, et al., Clin. Cancer Res. 18:3340–3351, 2012; I. Beyer, et al., Cancer Res. 71:7080–7090, 2011). Here, we show that affinity-enhanced versions of JO-1 are therapeutically more potent than the parental protein in a series of cancer models.
Canine Distemper Virus: an Emerging Disease in Wild Endangered Amur Tigers (
Panthera tigris altaica
)
Authors: Seimon Tracie A. , Miquelle Dale G. , Chang Tylis Y. , Newton Alisa L. , Korotkova Irina , Ivanchuk Galina , Lyubchenko Elena , Tupikov Andre , Slabe Evgeny , McAloose Denise 
Published: August 2013

ABSTRACT

Fewer than 500 Amur tigers (
Panthera tigris altaica
) remain in the wild. Due to low numbers and their solitary and reclusive nature, tiger sightings across their range in the Russian Far East and China are rare; sightings of sick tigers are rarer still. Serious neurologic disease observed in several wild tigers since 2001 suggested disease emergence in this endangered species. To investigate this possibility, histology, immunohistochemistry (IHC),
in situ
hybridization (ISH), and reverse transcription-PCR (RT-PCR) were performed on tissues from 5 affected tigers that died or were destroyed in 2001, 2004, or 2010. Our results reveal canine distemper virus (CDV) infection as the cause of neurologic disease in two tigers and definitively establish infection in a third. Nonsuppurative encephalitis with demyelination, eosinophilic nuclear viral inclusions, and positive immunolabeling for CDV by IHC and ISH were present in the two tigers with available brain tissue. CDV phosphoprotein (P) and hemagglutinin (H) gene products were obtained from brains of these two tigers by RT-PCR, and a short fragment of CDV P gene sequence was detected in lymph node tissue of a third tiger. Phylogenetically, Amur tiger CDV groups with an Arctic-like strain in Baikal seals (
Phoca siberica
). Our results, which include mapping the location of positive tigers and recognition of a cluster of cases in 2010, coupled with a lack of reported CDV antibodies in Amur tigers prior to 2000 suggest wide geographic distribution of CDV across the tiger range and recent emergence of CDV as a significant infectious disease threat to endangered Amur tigers in the Russian Far East.

IMPORTANCE
Recognition of disease emergence in wildlife is a rare occurrence. Here, for the first time, we identify and characterize a canine distemper virus (CDV), the second most common cause of infectious disease death in domestic dogs and a viral disease of global importance in common and endangered carnivores, as the etiology of neurologic disease and fatal encephalitis in wild, endangered Amur tigers. We establish that in 2010 CDV directly or indirectly killed ~1% of Amur tigers. Location of positive cases over an expansive geographic area suggests that CDV is widely distributed across the tiger range. Interspecies interactions are increasing as human populations grow and expand into wildlife habitats. Identifying animal reservoirs for CDV and identifying the CDV strains that are transmissible to and among wildlife species, including Amur tigers and sympatric critically endangered Amur leopards (
Panthera pardus orientalis
), is essential for guiding conservation and mitigation efforts.
The Activity and Specificity of the Outer Membrane Protein Chaperone SurA Are Modulated by a Proline Isomerase Domain
Authors: Ricci Dante P. , Schwalm Jaclyn , Gonzales-Cope Michelle , Silhavy Thomas J. 
Published: August 2013

ABSTRACT

SurA is a component of the periplasmic chaperone network that plays a central role in biogenesis of integral outer membrane β-barrel proteins (OMPs) in
Escherichia coli
. Although SurA contains two well-conserved proline isomerase (PPIase) domains, the contribution of these domains to SurA function is unclear. In the present work, we show that defects in OMP assembly caused by mutation of the β-barrel assembly factors BamA or BamB can be corrected by gain-of-function mutations in SurA that map to the first PPIase domain. These mutations apparently bypass the requirement for a stable interaction between SurA and the Bam complex and enhance SurA chaperone activity
in vivo
despite destabilization of the protein
in vitro
. Our findings suggest an autoinhibitory mechanism for regulation of SurA chaperone activity through interdomain interactions involving a PPIase domain. We propose a model in which SurA activity is modulated by an interaction between SurA and the Bam complex that alters the substrate specificity of the chaperone.

IMPORTANCE
The dominant
surA
mutations described here alter amino acid residues that are highly conserved in eukaryotic homologs of SurA, including Pin1, the human proline isomerase (PPIase) implicated in Alzheimer’s disease and certain cancers. Consequently, a mechanistic description of SurA function may enhance our understanding of clinically important PPIases and their role(s) in disease. In addition, the virulence of Gram-negative bacterial pathogens, such as
Salmonella
,
Shigella
, and
Escherichia coli
O157:H7, is largely dependent on SurA, making this PPIase/chaperone an attractive antibiotic target. Investigating the function of SurA in outer membrane (OM) biogenesis will be useful in the development of novel therapeutic strategies for the disruption of the OM or the processes that are essential for its assembly.
Does access to neighbourhood green space promote a healthy duration of sleep? Novel findings from a cross-sectional study of 259 319 Australians
Authors: Astell-Burt Thomas , Feng Xiaoqi , Kolt Gregory S 
Published: August 2013

ObjectivesExperiments demonstrate that exposure to parks and other ‘green spaces’ promote favourable psychological and physiological outcomes. As a consequence, people who reside in greener neighbourhoods may also have a lower risk of short sleep duration (<6 h). This is potentially important as short sleep duration is a correlate of obesity, chronic disease and mortality, but so far this hypothesis has not been previously investigated.DesignCross-sectional data analysis.SettingNew South Wales, Australia.ParticipantsThis study investigated whether neighbourhood green space was associated with a healthier duration of sleep (to the nearest hour) among 259 319 Australians who completed the 45 and Up Study baseline questionnaire between 2006 and 2009 inclusive.Primary and secondary outcome measuresMultinomial logit regression was used to investigate the influence of an objective measure of green space on categories of sleep duration: 8 h (normal); between 9 and 10 h (mid-long sleep); over 10 h (long sleep); between 6 and 7 h (mid-short sleep); and less than 6 h (short sleep). Models were adjusted for psychological distress, physical activity and a range of demographic and socioeconomic characteristics.ResultsPeople living in greener neighbourhoods reported a lower risk of short sleep. For example, compared with participants living in areas with 20% green space land-use, the relative risk ratios for participants with 80%+ green space was 0.86 (95% CI 0.81 to 0.92) for durations between 6 and 7 h, and 0.68 (95% CI 0.57 to 0.80) for less than 6 h sleep. Unexpectedly, the benefit of more green space for achieving 8 h of sleep was not explained by controls for psychological distress, physical activity or other socioeconomic factors.ConclusionsGreen space planning policies may have wider public health benefits than previously recognised. Further research in the role of green spaces in promoting healthier sleep durations and patterns is warranted.