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Kazuhiro Yamamoto, Salvatore Santamaria, Kenneth A Botkjaer, Jayesh Dudhia, Linda Troeberg, Yoshifumi Itoh, Gillian Murphy, Hideaki Nagase
OBJECTIVES: The aggrecanase ADAMTS-5 and the collagenase MMP-13 are constitutively secreted by chondrocytes in normal cartilage, but rapidly endocytosed via the cell surface endocytic receptor low-density lipoprotein receptor-related protein 1 (LRP1) and subsequently degraded. This endocytic system is impaired in osteoarthritic (OA) cartilage due to increased ectodomain 'shedding' of LRP1. The aim of this study is to identify the LRP1 sheddase(s) in human cartilage, and to test whether inhibition of LRP1 shedding prevents the cartilage degradation in OA...
February 24, 2017: Arthritis & Rheumatology
Chen Yan, Yaqing Chen, Wewei Kong, Liya Fu, Yunde Liu, Qingjuan Yao, Yuhua Yuan
Accumulating evidences indicate ectopic expression of noncoding RNAs are responsible for breast cancer progression. Increased non-coding RNA PVT1, the host gene of microRNA-1207-5p (miR-1207-5p), has been associated with breast cancer proliferation. However, how PVT1 functions in breast cancer is still not clear. In this study, we show a PVT1 derived microRNA, miR-1207-5p, which promotes the proliferation of breast cancer cells by directly regulating STAT6. We first confirm the positive correlated expression pattern between PVT1 and miR-1207-5p by observing consistent induced expression by estrogen, and overexpression in breast cancer cell lines and breast cancer patient specimens...
February 24, 2017: Cancer Science
Antonio C G Foddai, Irene R Grant
AIM: To validate an optimised Peptide-mediated magnetic separation (PMS)-Phage assay for detection of viable Mycobacterium avium subsp. paratuberculosis (MAP) in milk. METHODS AND RESULTS: Inclusivity, specificity and limit of detection 50% (LOD50 ) of the optimised PMS-phage assay were assessed. Plaques were obtained for all 43 MAP strains tested. Of 12 other Mycobacterium spp. tested, only M. bovis BCG produced small numbers of plaques. LOD50 of the PMS-phage assay was 0...
February 24, 2017: Journal of Applied Microbiology
Cory M Vela, Todd C Knepper, Nancy K Gillis, Christine M Walko, Howard L McLeod, J Kevin Hicks
INTRODUCTION: Moffitt Cancer Center's Personalized Medicine Clinical Service (PMCS) reviews somatic next-generation sequencing (NGS) assay results, provides interpretations, and identifies potential therapeutic options. The number of individuals reviewed by our clinical service who are eligible for on-label or off-label drug therapy based on genetic test results has previously not been quantitated. Herein, we determined the number of patients harboring an actionable mutation that would qualify a patient for an on-label drug or consideration for off-label drug treatment...
February 24, 2017: Pharmacotherapy
Samir Das, Sanghati Roy Chowdhury, Sanjay Dey, Udayaditya Sen
Vibrio cholerae experiences a highly hostile environment at human intestine which triggers the induction of various heat shock genes. The hchA gene product of V. cholerae O395, referred to a hypothetical intracellular protease/amidase VcHsp31, is one such stress-inducible homodimeric protein. Our current study demonstrates that VcHsp31 is endowed with molecular chaperone, amidopeptidase and robust methylglyoxalase activities. Through site directed mutagenesis coupled with biochemical assays on VcHsp31, we have confirmed the role of residues in the vicinity of the active site towards amidopeptidase and methylglyoxalase activities...
2017: PloS One
Giulia Breveglieri, Anna Travan, Elisabetta D'Aversa, Lucia Carmela Cosenza, Patrizia Pellegatti, Giovanni Guerra, Roberto Gambari, Monica Borgatti
The β-thalassemias are genetic disorder caused by more than 200 mutations in the β-globin gene, resulting in a total (β0) or partial (β+) deficit of the globin chain synthesis. The most frequent Mediterranean mutations for β-thalassemia are: β039, β+IVSI-110, β+IVSI-6 and β0IVSI-1. Several molecular techniques for the detection of point mutations have been developed based on the amplification of the DNA target by polymerase chain reaction (PCR), but they could be labor-intensive and technically demanding...
2017: PloS One
Lucila Okuyama Fukasawa, Maria Gisele Gonçalves, Fábio Takenori Higa, Euclides Ayres Castilho, Ana Belén Ibarz-Pavón, Claudio Tavares Sacchi
BACKGROUND: The lack of information regarding the burden of acute bacterial meningitis in Latin America leads to a reduction in the estimated incidence rates of the disease, and impairs public health decisions on the use and follow-up of preventive interventions, particularly, the evaluation of existing vaccination policies. The use of the real-time PCR in diagnostic routine procedures has resulted in a substantial increase in confirmed bacterial meningitis cases. However, in resource-poor countries, these assays are only available in reference laboratories...
2017: PloS One
Gamal M K Mehaisen, Mariam G Eshak, Ahmed M Elkaiaty, Abdel-Rahman M M Atta, Magdi M Mashaly, Ahmed O Abass
The massive meat production of broiler chickens make them continuously exposed to potential stressors that stimulate releasing of stress-related hormones like corticosterone (CORT) which is responsible for specific pathways in biological mechanisms and physiological activities. Therefore, this research was conducted to evaluate a wide range of responses related to broiler performance, immune function, plasma biochemistry, related gene expressions and cell death morphology during and after a 7-day course of CORT injection...
2017: PloS One
Maria A Greenwood, Elizabeth A D Hammock
Oxytocin (OXT) is a pleiotropic regulator of physiology and behavior. An emerging body of evidence demonstrates a role for OXT in the transition to postnatal life of the infant. To identify potential sites of OXT action via the OXT receptor (OXTR) in the newborn mouse, we performed receptor autoradiography on 20 μm sagittal sections of whole postnatal day 0 male and female mice on a C57BL/6J background using the 125iodinated ornithine vasotocin analog ([125I]-OVTA) radioligand. A competitive binding assay on both wild-type (WT) and OXTR knockout (OXTR KO) tissue was used to assess the selectivity of [125I]-OVTA for neonatal OXTR...
2017: PloS One
Hiroki Kanazawa, Keisuke Imoto, Muneyoshi Okada, Hideyuki Yamawaki
A hypoxic stress which causes apoptosis of cardiomyocytes is the main problem in the ischemic heart disease. Canstatin, a non-collagenous fragment of type IV collagen α2 chain, is an endogenous anti-angiogenic factor. We have previously reported that canstatin has a cytoprotective effect on cardiomyoblasts. In the present study, we examined the effects of canstatin on hypoxia-induced apoptosis in H9c2 cardiomyoblasts. Cell counting assay was performed to determine a cell viability. Western blotting was performed to detect expression of cleaved casepase-3 and phosphorylation of focal adhesion kinase (FAK) and Akt...
2017: PloS One
Marisa Simon, Fahmi Mesmar, Luisa Helguero, Cecilia Williams
Triple-negative breast cancer (TNBC) is an aggressive, highly recurrent breast cancer subtype, affecting approximately one-fifth of all breast cancer patients. Subpopulations of treatment-resistant cancer stem cells within the tumors are considered to contribute to disease recurrence. A potential druggable target for such cells is the maternal embryonic leucine-zipper kinase (MELK). MELK expression is upregulated in mammary stem cells and in undifferentiated cancers, where it correlates with poor prognosis and potentially mediates treatment resistance...
2017: PloS One
Dale B Bosco, Mark D Roycik, Yonghao Jin, Martin A Schwartz, Ty J Lively, Diego A R Zorio, Qing-Xiang Amy Sang
Development of adipose tissue requires the differentiation of less specialized cells, such as human mesenchymal stem cells (hMSCs), into adipocytes. Since matrix metalloproteinases (MMPs) play critical roles in the cell differentiation process, we conducted investigations to determine if a novel mercaptosulfonamide-based MMP inhibitor (MMPI), YHJ-7-52, could affect hMSC adipogenic differentiation and lipid accumulation. Enzyme inhibition assays, adipogenic differentiation experiments, and quantitative PCR methods were employed to characterize this inhibitor and determine its effect upon adipogenesis...
2017: PloS One
Kathrin Schalk, Barbara Lexhaller, Peter Koehler, Katharina Anne Scherf
Gluten proteins from wheat, rye, barley and, in rare cases, oats, are responsible for triggering hypersensitivity reactions such as celiac disease, non-celiac gluten sensitivity and wheat allergy. Well-defined reference materials (RM) are essential for clinical studies, diagnostics, elucidation of disease mechanisms and food analyses to ensure the safety of gluten-free foods. Various RM are currently used, but a thorough characterization of the gluten source, content and composition is often missing. However, this characterization is essential due to the complexity and heterogeneity of gluten to avoid ambiguous results caused by differences in the RM used...
2017: PloS One
Xu Chen, Haiyan Ye, Shilin Li, Baihai Jiao, Jianqin Wu, Peibin Zeng, Limin Chen
Severe fever with thrombocytopenia syndrome (SFTS) is an emerging infectious disease caused by a novel bunyavirus (SFTS virus, SFTSV). At present there is still no specific antiviral treatment for SFTSV; To understand which cells support SFTSV life cycle and whether SFTSV infection activates host innate immunity, four different cell lines (Vero, Hela, Huh7.5.1, and Huh7.0) were infected with SFTSV. Intracellular/extracellular viral RNA and expression of IFNα, and IFNß were detected by real-time RT- PCR following infection...
2017: PloS One
Chandra Shekhar Pareek, Paweł Błaszczyk, Piotr Dziuba, Urszula Czarnik, Leyland Fraser, Przemysław Sobiech, Mariusz Pierzchała, Yaping Feng, Haja N Kadarmideen, Dibyendu Kumar
BACKGROUND: RNA-seq is a useful next-generation sequencing (NGS) technology that has been widely used to understand mammalian transcriptome architecture and function. In this study, a breed-specific RNA-seq experiment was utilized to detect putative single nucleotide polymorphisms (SNPs) in liver tissue of young bulls of the Polish Red, Polish Holstein-Friesian (HF) and Hereford breeds, and to understand the genomic variation in the three cattle breeds that may reflect differences in production traits...
2017: PloS One
Jose V Die, Ransom L Baldwin, Lisa J Rowland, Robert Li, Sunghee Oh, Congjun Li, Erin E Connor, Maria-Jose Ranilla
The rumen is lined on the luminal side by a stratified squamous epithelium that is responsible for not only absorption, but also transport, extensive short-chain fatty acid (SCFA) metabolism and protection. Butyrate has been demonstrated to initiate the differentiation of the tissue following introduction of solid feed to the weaning neonate as well as affecting the metabolism of other nutrients and absorption of nutrients in in vitro experiments. The objective of the present study was to validate expression stability of eight putative reference genes bovine rumen, considering the intrinsic heterogeneity of bovine rumen with regard to different luminal characteristics due to direct infusion of butyrate to double the intra-ruminal content of the rumen liquor...
2017: PloS One
Marina A Afanasyeva, Lidia V Putlyaeva, Denis E Demin, Ivan V Kulakovskiy, Ilya E Vorontsov, Marina V Fridman, Vsevolod J Makeev, Dmitry V Kuprash, Anton M Schwartz
We studied functional effect of rs12722489 single nucleotide polymorphism located in the first intron of human IL2RA gene on transcriptional regulation. This polymorphism is associated with multiple autoimmune conditions (rheumatoid arthritis, multiple sclerosis, Crohn's disease, and ulcerative colitis). Analysis in silico suggested significant difference in the affinity of estrogen receptor (ER) binding site between alternative allelic variants, with stronger predicted affinity for the risk (G) allele. Electrophoretic mobility shift assay showed that purified human ERα bound only G variant of a 32-bp genomic sequence containing rs12722489...
2017: PloS One
Cristina Carranza-Rodríguez, Daniel San-Román-Sánchez, Héctor Marrero-Santiago, Michele Hernández-Cabrera, Carlos Gil-Guillén, Elena Pisos-Álamo, Nieves Jaén-Sánchez, José-Luis Pérez-Arellano
BACKGROUND: Among immigrants of sub-Saharan origin, parasitic infection is the leading cause of eosinophilia, which is generally interpreted as a defense mechanism. A side effect of the inflammatory mediators released by eosinophils is damage to host organs, especially the heart. The main objectives of this study were to i) assess cardiac involvement in asymptomatic sub-Saharan immigrants with eosinophilia, ii) relate the presence of lesions with the degree of eosinophilia, and iii) study the relationship between cardiac involvement and the type of causative parasite...
February 24, 2017: PLoS Neglected Tropical Diseases
Palaniyandi Karuppaiya, Xiao-Xue Yan, Wang Liao, Jun Wu, Fang Chen, Lin Tang
Physic nut (Jatropha curcas L) seed oil is a natural resource for the alternative production of fossil fuel. Seed oil production is mainly depended on seed yield, which was restricted by the low ratio of staminate flowers to pistillate flowers. Further, the mechanism of physic nut flower sex differentiation has not been fully understood yet. Quantitative Real Time-Polymerase Chain Reaction is a reliable and widely used technique to quantify the gene expression pattern in biological samples. However, for accuracy of qRT-PCR, appropriate reference gene is highly desirable to quantify the target gene level...
2017: PloS One
Yani Zhang, Yingjie Wang, Qisheng Zuo, Dong Li, Wenhui Zhang, Fei Wang, Yanqin Ji, Jing Jin, Zhenyu Lu, Man Wang, Chen Zhang, Bichun Li
An efficient genome editing approach had been established to construct the stable transgenic cell lines in the domestic chicken (Gallus gallus domesticus) at present. Our objectives were to investigate gene function in the differentiation process of chicken embryonic stem cells (ESCs) into spermatogonial stem cells(SSCs). Three guides RNA (gRNAs) were designed to knockout the Stra8 gene, and knockout efficiency was evaluated in domestic chicken cells using cleavage activity of in vitro transcription of gRNA, Luciferase-SSA assay, T7 endonuclease I assay(T7E1) and TA clone sequence...
2017: PloS One
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