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https://www.readbyqxmd.com/read/28643557/pooled-sample-testing-for-bonamia-ostreae-a-tale-of-two-sybr-green-real-time-pcr-assays
#1
Henry S Lane, J Brian Jones, Wendy L McDonald
Pooled testing of samples is a common laboratory practice to increase efficiency and reduce expenses. We investigated the efficacy of 2 published SYBR Green real-time PCR assays when used to detect the haplosporidian parasite Bonamia ostreae in pooled samples of infected oyster tissue. Each PCR targets a different gene within the B. ostreae genome: the actin 1 gene or the 18S rRNA gene. Tissue homogenates (150 mg) of the New Zealand flat oyster Ostrea chilensis were spiked with ~1.5 × 10(3) purified B. ostreae cells to create experimental pools of 3, 5, and 10...
June 1, 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28643089/time-lapse-observation-and-transcriptome-analysis-of-a-case-with-repeated-multiple-pronuclei-after-ivf-icsi
#2
J Dai, L Z Leng, C F Lu, F Gong, S P Zhang, W Zheng, G X Lu, G Lin
PURPOSE: The purpose of this study was to investigate the cause of repeated multipronucleus (MPN) formation in zygotes in a patient after both in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI). METHOD: This is a case study. A patient had unexplained primary infertility with recurring total MPN zygotes after IVF and ICSI cycles. Time-lapse monitoring of pronucleus formation was carried out. Embryos developed from MPN zygotes were analyzed by fluorescence in situ hybridization (FISH)...
June 22, 2017: Journal of Assisted Reproduction and Genetics
https://www.readbyqxmd.com/read/28640195/pharmacogenomic-variants-may-influence-the-urinary-excretion-of-novel-kidney-injury-biomarkers-in-patients-receiving-cisplatin
#3
Cara Chang, Yichun Hu, Susan L Hogan, Nickie Mercke, Madeleine Gomez, Cindy O'Bryant, Daniel W Bowles, Blessy George, Xia Wen, Lauren M Aleksunes, Melanie S Joy
Nephrotoxicity is a dose limiting side effect associated with the use of cisplatin in the treatment of solid tumors. The degree of nephrotoxicity is dictated by the selective accumulation of cisplatin in renal tubule cells due to: (1) uptake by organic cation transporter 2 (OCT2) and copper transporter 1 (CTR1); (2) metabolism by glutathione S-transferases (GSTs) and γ-glutamyltransferase 1 (GGT1); and (3) efflux by multidrug resistance-associated protein 2 (MRP2) and multidrug and toxin extrusion protein 1 (MATE1)...
June 22, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28639054/a-strong-relationship-between-oral-squamous-cell-carcinoma-and-dna-repair-genes
#4
Hakan Avci, Arzu Ergen, Elif Sinem Bireller, Baris Ertugrul, Bedia Cakmakoglu
Single nucleotide polymorphisms of DNA repair genes alter protein function and modulate DNA repair efficiency in various cancers. The X-ray repair cross-complementing group (XRCC) is responsible for the repair of DNA base damage and single-strand breaks. The aim of our study was to investigate the association of XRCC1 Arg399Gln and XRCC3 Thr241Met polymorphisms with the susceptibility to develop oral squamous cell carcinoma (OSCC) in Turkish subjects. One hundred eleven patients with OSCC and 148 healthy controls were recruited for the study...
June 21, 2017: Biochemical Genetics
https://www.readbyqxmd.com/read/28637324/the-effects-of-phytosterols-on-the-sexual-behavior-and-reproductive-function-in-the-japanese-quail-coturnix-coturnix-japonica
#5
Mohammad Ibrahim Qasimi, Kentaro Nagaoka, Gen Watanabe
Phytosterols (PS) are plant origin sterols naturally found in many foods and added as food additives. Since 1950, PS have attracted considerable attention due to their ability to lower serum cholesterol and inhibit cardiovascular disease (CVD). However, recent studies have found that PS act as endocrine-disrupting chemicals in laboratory animals. Therefore, this study was aimed at finding the mechanism(s) for PS effects on the sexual behaviors and reproductive functions in male Japanese quails (Coturnix coturnix japonica)...
June 16, 2017: Poultry Science
https://www.readbyqxmd.com/read/28632844/complement-mediated-regulation-of-apolipoprotein-e-in-cultured-human-rpe-cells
#6
Ping Yang, Nikolai P Skiba, Grace M Tewkesbury, Victoria M Treboschi, Peter Baciu, Glenn J Jaffe
PURPOSE: Complement activation is implicated in the pathogenesis of age-related macular degeneration (AMD). Apolipoprotein E (ApoE) and complement activation products such as membrane attack complex (MAC) are present in eyes of individuals with AMD. Herein, we investigated the effect of complement activation on induction of ApoE accumulation in human retinal pigment epithelial (RPE) cells. METHODS: Cultured human RPE cells were primed with a complement-fixing antibody followed by treatment with C1q-depleted (C1q-Dep) human serum to elicit alternative pathway complement activation...
June 1, 2017: Investigative Ophthalmology & Visual Science
https://www.readbyqxmd.com/read/28629734/immunohistochemical-and-genetic-exploration-of-incompatible-a-blood-group-antigen-expression-in-invasive-micropapillary-breast-carcinoma-a-case-report
#7
S Zouine, Z Orfi, K Kojok, S Benayad, Y Zaid, F Marnissi, N Habti
INTRODUCTION: Invasive Micropapillary Carcinoma (IMPC) of the breast is a relatively rare subtype of invasive ductal carcinoma and represents the most inherently aggressive form. Expression of incompatible blood group A antigen in cancer of type O patients has been reported in several types of cancer, however, the biosynthetic mechanism and the genetic basis remain unclear until today. The aim of the present case report study was to evaluate the expression of incompatible blood group A antigen and to identify the genetic basis of this expression in IMPC of the breast...
June 16, 2017: Current Research in Translational Medicine
https://www.readbyqxmd.com/read/28626778/single-cell-based-vector-tracing-in-patients-with-ada-scid-treated-with-stem-cell-gene-therapy
#8
Yuka Igarashi, Toru Uchiyama, Tomoko Minegishi, Sirirat Takahashi, Nobuyuki Watanabe, Toshinao Kawai, Masafumi Yamada, Tadashi Ariga, Masafumi Onodera
Clinical improvement in stem cell gene therapy (SCGT) for primary immunodeficiencies depends on the engraftment levels of genetically corrected cells, and tracing the transgene in each hematopoietic lineage is therefore extremely important in evaluating the efficacy of SCGT. We established a single cell-based droplet digital PCR (sc-ddPCR) method consisting of the encapsulation of a single cell into each droplet, followed by emulsion PCR with primers and probes specific for the transgene. A fluorescent signal in a droplet indicates the presence of a single cell carrying the target gene in its genome, and this system can clearly determine the ratio of transgene-positive cells in the entire population at the genomic level...
September 15, 2017: Molecular Therapy. Methods & Clinical Development
https://www.readbyqxmd.com/read/28624618/a-sample-to-answer-real-time-convective-polymerase-chain-reaction-system-for-point-of-care-diagnostics
#9
Bowen Shu, Chunsun Zhang, Da Xing
Timely and accurate molecular diagnostics at the point-of-care (POC) level is critical to global health. To this end, we propose a handheld convective-flow real-time polymerase chain reaction (PCR) system capable of direct sample-to-answer genetic analysis for the first time. Such a system mainly consists of a magnetic bead-assisted photothermolysis sample preparation, a closed-loop convective PCR reactor, and a wireless video camera-based real-time fluorescence detection. The sample preparation exploits the dual functionality of vancomycin-modified magnetic beads (VMBs) for bacteria enrichment and photothermal conversion, enabling cell pre-concentration and lysis to be finished in less than 3min...
June 8, 2017: Biosensors & Bioelectronics
https://www.readbyqxmd.com/read/28624489/association-of-genetic-polymorphisms-of-macrophage-inhibitory-factor-mif-and-b-cell-activating-factor-baff-with-the-detection-of-donor-specific-antibodies-in-kidney-allograft-recipients
#10
Youngil Chang, Tariq Shah, David I Min
The posttransplant development of donor specific antibodies (DSA) initiates the antibody mediated rejection (AMR), which is associated with the increased rate of graft loss. One of the characteristics of AMR is the infiltration of innate immune system including macrophages, monocytes, neutrophils or NK cells. Macrophage inhibitory factor (MIF) and B-cell activating factor (BAFF) are well known cytokines that are associated with the activation of the innate immune system which can damage kidney allograft. In this article, the association of the genetic polymorphisms of MIF and BAFF with the development of DSA including Class I and II in kidney transplant patients is investigated...
June 14, 2017: Human Immunology
https://www.readbyqxmd.com/read/28623657/aptamer-magnetic-bead-quantum-dot-sandwich-assays-for-foodborne-pathogen-detection-pros-cons-and-lessons-learned
#11
John G Bruno, Jeffrey C Sivils, Taylor Phillips
DNA and RNA aptamers have been extensively investigated as potential competitors for antibodies for a variety of applications including food safety testing. Ultrasensitive fluorescence detection of foodborne pathogenic bacteria as low as 1-10 cells/mL has been achieved using aptamers coupled to quantum dots in clear pristine buffers for environmental sample detection. Quantum dots offer other advantages, including single UV or blue light source multiplex (multicolored) detection. However, quantum dots can exhibit decreased fluorescence in some food matrixes and even completely fail to fluoresce in some fatty matrixes, as documented in this report...
June 16, 2017: Journal of AOAC International
https://www.readbyqxmd.com/read/28623396/assessment-of-the-circulating-cell-free-dna-marker-association-with-diagnosis-and-prognostic-prediction-in-patients-with-lymphoma-a-single-center-experience
#12
Mao Li, Yongqian Jia, Juan Xu, Xiaomin Cheng, Caigang Xu
Circulating cell-free DNA (ccfDNA) has been shown to be associated with the clinical characteristics and prognosis of cancer patients. Our objective was to assess whether the concentration and integrity index of ccfDNA in plasma may be useful for diagnosing and monitoring the progression of patients with lymphoma. We included plasma samples from 174 lymphoma patients and 80 healthy individuals. The total concentration of ccfDNA was determined using a fluorometry method, and the DNA integrity index (DII), which is the ratio of longer to shorter DNA fragments, for the APP gene was detected using real-time quantitative PCR...
June 16, 2017: Annals of Hematology
https://www.readbyqxmd.com/read/28623130/leukotriene-signaling-via-alox5-and-cysteinyl-leukotriene-receptor-1-is-dispensable-for-in%C3%A2-vitro-growth-of-cd34-cd38-stem-and-progenitor-cells-in-chronic-myeloid-leukemia
#13
Monika Dolinska, Alexandre Piccini, Wan Man Wong, Eleni Gelali, Anne-Sofie Johansson, Johannis Klang, Pingnan Xiao, Elham Yektaei-Karin, Ulla Olsson Strömberg, Satu Mustjoki, Leif Stenke, Marja Ekblom, Hong Qian
Tyrosine kinase inhibitors targeting the BCR-ABL oncoprotein in chronic myeloid leukemia (CML) are remarkably effective inducing deep molecular remission in most patients. However, they are less effective to eradicate the leukemic stem cells (LSC), resulting in disease persistence. Therefore, there is great need to develop novel therapeutic strategies to specifically target the LSC. In an experimental mouse CML model system, the leukotriene pathway, and specifically, the expression ALOX5, encoding 5-lipoxygenase (5-LO), has been reported as a critical regulator of the LSC...
June 13, 2017: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/28622672/physiological-effects-of-single-and-multi-walled-carbon-nanotubes-on-rice-seedlings
#14
Hao Zhang, Mengxia Yue, Xueke Zheng, Chengshen Xie, Hong Zhou, Lijia Li
In this paper, single-walled carbon nanotubes (SWCNTs) and multi-walled carbon nanotubes (MWCNTs) were found to accelerate leaf growth and development of rice seedlings at a low concentration (20 mg/L), accompanied with the increased chlorophyll content and net photosynthetic rate (PN). Quantitative real-time polymerase chain reaction (RT-PCR) results indicated that both SWCNTs and MWCNTs significantly increased expression of genes associated with chloroplast development and cell sizes. Further analysis revealed that the abscisic acid (ABA) content decreased and the gibberellin (GA) content increased while the content of O2 and H2O2 was slightly elevated and the activities of antioxidative enzymes (SOD, EC 1...
June 14, 2017: IEEE Transactions on Nanobioscience
https://www.readbyqxmd.com/read/28618935/heterogeneous-dna-methylation-status-in-same-cell-subpopulations-of-ovarian-cancer-tissues
#15
Qiling Li, Xue Xue, Wenzhi Li, Qi Wang, Lu Han, Tiffany Brunson, Wei Xu, Isfahan Chambers-Harris, Qing Wang, Xu Li, Li Ma, Qing Song
This study aims to explore the heterogeneous DNA methylation differences between individual single ovarian cancer cells isolated from the same formalin-fixed and paraffin-embedded human ovarian cancer tissue. Single cells were isolated by laser microdissection. Whole genome amplification and polymerase chain reaction purification were performed on the converted genomic DNA. Target primers designed for checking DNA methylation were used in polymerase chain reaction reactions to amplify special fragments. Sequencing was performed to analyze the heterogeneous DNA methylation statuses of different single ovarian cancer cells...
June 2017: Tumour Biology: the Journal of the International Society for Oncodevelopmental Biology and Medicine
https://www.readbyqxmd.com/read/28618172/dominance-of-sulfuritalea-species-in-nitrate-depleted-water-of-a-stratified-freshwater-lake-and-arsenate-respiration-ability-within-the-genus
#16
Tomohiro Watanabe, Aya Miura, Tomoya Iwata, Hisaya Kojima, Manabu Fukui
Facultatively autotrophs of the genus Sulfuritalea within the class Betaproteobacteria have been predicted to be an important bacterial population in stratified freshwater lakes based on previous PCR-based studies. Here, we designed a new probe specific for the genus Sulfuritalea and performed catalyzed reporter deposition-fluorescence in situ hybridization to enumerate cells of Sulfuritalea species throughout the water column in a stratified freshwater lake. The cells stained with the Sulfuritalea-specific probe were detected in all hypoxic water samples collected in different seasons and years...
June 15, 2017: Environmental Microbiology Reports
https://www.readbyqxmd.com/read/28618041/effect-of-various-ratios-of-co-cultured-atdc5-cells-and-chondrocytes-on-the-expression-of-cartilaginous-phenotype-in-microcavitary-alginate-hydrogel
#17
Yongchang Yao, Yuyang Huang, Dongyang Qian, Shujiang Zhang, Yi Chen, Bo Bai
The present study introduced a direct co-culture of mouse ATDC5 cells and primary porcine chondrocytes into a microcavitary hydrogel, which possessed advantages in promoting the growth of chondrocytes and retaining the phenotype. These two types of cells were encapsulated with gelatin microspheres in alginate hydrogels in either of the three ratios (3:1, 1:1, or 1:3 of ATDC5 cells to chondrocytes) and cultured in chondrogenic medium for 28 days. Simultaneously, the single encapsulation of ATDC5 cells or chondrocytes was set as a control...
June 15, 2017: Journal of Cellular Biochemistry
https://www.readbyqxmd.com/read/28617574/synergistic-suppression-of-tumor-angiogenesis-by-co-delivering-of-vegf-targeted-sirna-and-candesartan-mediated-by-functionalized-carbon-nanovectors
#18
Xuefang Ding, Yujie Su, Cheng Wang, Fangrong Zhang, Kerong Chen, Yu Wang, Min Li, Wei Wang
Single-walled carbon nanotubes (SWNTs) with unique physicochemical properties have exhibited promising biomedical applications as drug/gene carrier. In this study, polyethyleneimine (PEI) modified SWNT conjugates linked with candesartan (CD) were developed to deliver vascular endothelial growth factor (VEGF) targeted siRNA (siVEGF) for synergistic and targeted treatment of tumor angiogenesis. The characterization results revealed that SWNT-PEI-CD conjugates were successfully synthesized, and exhibited desirable dispersibility and superior stability...
June 15, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28617560/influence-of-vegf-bmp-2-on-the-proliferation-and-osteogenetic-differentiation-of-rat-bone-mesenchymal-stem-cells-on-plga-gelatin-composite-scaffold
#19
G An, W-B Zhang, D-K Ma, B Lu, G-J Wei, Y Guang, C-H Ru, Y-S Wang
OBJECTIVE: To investigate the influence of VEGF/BMP-2 on the proliferation and osteogenic differentiation of rat bone mesenchymal stem cells BMSCs) on PLGA/gelatin composite scaffold. MATERIALS AND METHODS: Randomly-oriented nanofibers with different ratios of Poly Lactic-co-Glycolic Acid (PLGA)/gelatin were produced through electrospinning. The mixture of nanofibers and BMSCs was pipetted onto the surface of the scaffolds, and BMSCs/PLGA/gelatin composite was obtained...
May 2017: European Review for Medical and Pharmacological Sciences
https://www.readbyqxmd.com/read/28616967/effects-of-functional-groups-of-materials-on-nonspecific-adhesion-and-chondrogenic-induction-of-mesenchymal-stem-cells-on-free-and-micropatterned-surfaces
#20
Bin Cao, Yuanmeng Peng, Xiangnan Liu, Jiandong Ding
Functional groups of materials are known to affect cell behaviors, yet the corresponding effect on stem cell differentiation are always coupled with that of cell spreading, it is thus unclear whether or not the chemical groups influence cell differentiation directly or via cell spreading indirectly. Herein we used a unique surface patterning technique to decouple the corresponding effects. Mesenchymal stem cells (MSCs) derived from bone marrow were seeded on surfaces coated with alkylthiols with one of 4 functional end groups (-CH3, -OH, -COOH and -NH2) and underwent 9-day chondrogenic induction...
June 15, 2017: ACS Applied Materials & Interfaces
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