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cytometri of flow

Kristýna Hanušová, Martin Certner, Tomáš Urfus, Petr Koutecký, Jirí Košnar, Carl J Rothfels, Vlasta Jarolímová, Jan Ptácek, Libor Ekrt
Background and Aims: Polyploidy has played an important role in the evolution of ferns. However, the dearth of data on cytotype diversity, cytotype distribution patterns and ecology in ferns is striking in comparison with angiosperms and prevents an assessment of whether cytotype coexistence and its mechanisms show similar patterns in both plant groups. Here, an attempt to fill this gap was made using the ploidy-variable and widely distributed Cystopteris fragilis complex. Methods: Flow cytometry was used to assess DNA ploidy level and monoploid genome size (Cx value) of 5518 C...
December 12, 2018: Annals of Botany
Felix Umrath, Carla Thomalla, Simone Pöschel, Katja Schenke-Layland, Siegmar Reinert, Dorothea Alexander
BACKGROUND/AIMS: Periosteal tissue is a valuable source of multipotent stem cells for bone tissue engineering. To characterize these cells in detail, we generated an immortalized human cranial periosteal cell line and observed an increased MSCA-1 and CD146 expression, as well as an earlier and stronger mineralization compared to the parental cells. Further, we detected a higher osteogenic potential of MSCA-1high compared to MSCA-1low cranial periosteal cell (CPC) fractions. In the present study, a possible synergism of MSCA-1 and CD146 for periosteal cell mineralization was investigated...
December 12, 2018: Cellular Physiology and Biochemistry
Rong-Fu Chen, Chih-Hau Chang, Chun-Ting Wang, Yur-Ren Kuo
BACKGROUND: Oxygen free radicals play a central role in diabetic angiopathy. This study investigated whether suppression of oxygen radicals could decrease endothelial damage and increase peripheral tissue circulation in a diabetic rodent model. METHODS: Sprague-Dawley rats were treated using streptozotocin to induce diabetes. The experiments were performed 4 weeks after diabetes induction: group 1: control, consisted of normal rats; group 2: diabetes, did not receive treatment; groups III (SOD10) and IV (SOD50): diabetes, received polyethylene glycol-conjugated superoxide dismutase (SOD), an antioxidant, 10 and 50 U/kg per day intraperitoneally for 4 weeks...
January 2019: Annals of Plastic Surgery
Hara Levy, Shuang Jia, Amy Pan, Xi Zhang, Mary L Kaldunski, Melodee L Nugent, Melissa Reske, Rachel A Feliciano, Diana Quintero, Michael M Renda, Katherine J Woods, Kathy Murkowski, Keven Johnson, James Verbsky, Trivikram Dasu, Justin Eze Ideozu, Susanna McColley, Michael W Quasney, Mary K Dahmer, Ellis D Avner, Philip M Farrell, Carolyn L Cannon, Howard Jacob, Pippa M Simpson, Martin J Hessner
Although cystic fibrosis (CF) is attributed to dysfunction of a single gene, the relationships between the abnormal gene product and development of inflammation and progression of lung disease are not fully understood, limiting our ability to predict an individual patient's clinical course and treatment response. To better understand CF progression, we characterized the molecular signatures of CF disease status using plasma-based functional genomics. Peripheral blood mononuclear cells (PBMCs) from healthy donors were cultured with plasma samples from CF patients (n=103) and unrelated, healthy controls (n=31)...
December 12, 2018: Physiological Genomics
Yan Ma, Xiaojing Jia
BACKGROUND Exposure to ionizing radiation (IR) induces severe damage in multiple human tissues. The testes are extremely sensitive to IR, and testes irradiation can result in infertility and abnormality. A novel and safe radioprotector for testes injury from IR is needed. Polydatin (PD) has been proved to have anti-oxidant and anti-inflammatory effects, indicating its potential application in radiation protection. MATERIAL AND METHODS Male wild-type C57BL/6 mice (8 weeks old) were exposed to ionizing radiation...
December 12, 2018: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
Hu Liu, Ming-Rui Xie, Hui Gao, Jian Li
Aims: Breast cancer is a great public health problem. It remains unclear how pharmorubicin induces cardiac dysfunction in patients with breast cancer. Our study was aimed to explore the influence of pharmorubicin on the left ventricular ejection fraction (EF) of patients with breast cancer and its potential mechanism. Materials and Methods: Patients with breast cancer were enrolled at the same hospital. Group I consisted of 135 samples, who were under treatment of pharmorubicin (200 mg/m2 )...
December 2018: Journal of Cancer Research and Therapeutics
Chengkai Xu, Hanbin Guo, Danli Kong, Dong Pang, Yuanlin Ding
Context: The metastasis of liver cancer is a major cause of clinical treatment failure, restrain, and control the cancer metastasis is the major strategy of the treatment and prevention of the disease. Special AT-rich sequence-binding protein 1 (SATB1) gene was overexpressed in many malignant tumors and considered as a potential target of anticancer drug. This study investigated the mechanism how ganodermanontriol effect the expression of SATB1 and thus inhibits the growth and metastasis in hepatocellular carcinoma (HCC)...
December 2018: Journal of Cancer Research and Therapeutics
Lijun Ye, Jimei Zhou, Wei Zhao, Pengfei Jiao, Gaofei Ren, Shujun Wang
Aim of the Study: Garcinia hanburyi is a traditional herbal medicine with activities of anti-inflammation and hemostasis used by people in South Asia. Gambogic acid (GA) is the main active component extracted from it, which has anticancer and anti-inflammatory effects. The aim of the current study is to investigate the molecular mechanisms of GA's effective anticancer activity. Materials and Methods: 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay was used to measure cell proliferation...
December 2018: Journal of Cancer Research and Therapeutics
Dan-Dan Xu, Wen-Ting Li, Dan Jiang, Huai-Guo Wu, Ming-Shan Ren, Mei-Qiao Chen, Yuan-Bo Wu
A high glucose state readily causes peripheral axon atrophy, demyelination, loss of nerve fiber function, and delayed regeneration. However, few studies have examined whether nitration is also critical for diabetic peripheral neuropathy. Therefore, this study investigated the effects of high glucose on proliferation, apoptosis, and 3-nitrotyrosine levels of Schwann cells treated with butylphthalide. In addition, we explored potential protective mechanisms of butylphthalide on peripheral nerves. Schwann cells were cultured in vitro with high glucose then stimulated with the peroxynitrite anion inhibitors uric acid and 3-n-butylphthalide for 48 hours...
March 2019: Neural Regeneration Research
Fei Cao, Li-Xue Yin
BACKGROUND: Hepatocellular carcinoma (HCC) is one of the primary causes of cancer-related death and resistance to cytotoxic chemotherapy is the major cause of mortality in HCC patients. miR-122 is a liver specific miRNA and is found to be reduced in HCC, however, the function of miR-122 in HCC chemosensitivity remains elusive. METHODS: We used qRT-PCR to measure expressions of miR-122, β-catenin and MDR1 in four HCC cell lines. And we assessed the effects of miR-122 or β-catenin on cell viability and apoptosis upon oxaliplatin (OXA) treatment by MTT assay and flow cytometry...
October 26, 2018: Experimental and Molecular Pathology
Paramita Ghosh, Debarpan Mitra, Sreyashi Mitra, Sudipta Ray, Samir Banerjee, Nabendu Murmu
BACKGROUND: Madhuca indica belongs to the family sapotaceae, commonly known as Mahua. It is primarily known for alcoholic beverage production and is reported to have anti-inflammatory, analgesic and antipyretic properties. Madhuca indica has also been reported to be effective in several diseases. OBJECTIVE: This study was undertaken to check the anticancer efficacy and chemopreventive effect of methanolic extract of Mahua flower (ME) on human breast cancer cell lines MCF-7 and MDA-MB-468...
December 11, 2018: Current Molecular Medicine
Fortunato Ferrara, Maria Felicia Soluri, Daniele Sblattero
In vitro display technologies have put together the generation of large antibody libraries with selection and screening procedures to identify lead candidates. Phage display antibody libraries allow selecting and identifying binders for a variety of antigens. Nonetheless, the procedure is limited by the possibility to quantitatively follow the enrichment during selection cycles and tune up the clones for specific binding proprieties (i.e., affinity). Yeast display allows the expression of thousands of copies of the antibody on each cell, simultaneously carrying the plasmid encoding that antibody, moreover the selection parameters can be accurately controlled by flow cytometry-based analysis and sorting...
2019: Methods in Molecular Biology
Keichiro Mihara, Tetsumi Yoshida, Joyeeta Bhattacharyya
Chimeric antigen receptors against CD19 (anti-CD19-CAR) are widely recognized and used by not only researchers associated with immunology, molecular biology, and cell biology but also physicians to treat B-cell malignancies. Anti-CD19-CAR is currently clinically available as one of the therapeutic modalities for refractory acute B-cell-typed lymphoblastic leukemia (B-ALL) patients. However, to detect CAR on the cell surface and investigate the efficacy of CAR-T cells, there are numerous experimental modalities including flow cytometry, the Cr-releasing assay, immunoblot, and immunostaining...
2019: Methods in Molecular Biology
Jeremy D King, John C Williams
Monoclonal antibodies (mAbs) are a major therapeutic modality. Grafting the meditope binding site onto mAbs, also known as meditope-enabling, can extend the usefulness of mAbs by providing an additional protein-protein interaction surface without altering the stability or antigen binding. We have previously used this site for attaching dyes, cytotoxic drugs, and entire proteins. Here, we provide a simple protocol for meditope-enabling mAbs, and verifying meditope and antigen binding using flow cytometry (FACS)...
2019: Methods in Molecular Biology
Anna-Jasmina Donaubauer, Paul F Rühle, Ina Becker, Rainer Fietkau, Udo S Gaipl, Benjamin Frey
Recent improvements in the flow cytometry technology allow the determination of the general immune status through the development of multicolor immunofluorescence panels. The one-tube multicolor flow cytometry assay (OTMA) that is presented here identifies 20 different, clinically relevant immune cell subsets and three common activation markers. Thereby, a comprehensive immune status that covers all major immune cells is easily obtained.The assay is suitable for every common three lasers and 10 color flow cytometer and includes the application of 15 different antibodies...
2019: Methods in Molecular Biology
Renata L Markman, Liana P Webber, Carlos H V Nascimento Filho, Leonardo A Reis, Pablo A Vargas, Marcio A Lopes, Virgilio Zanella, Manoela D Martins, Cristiane H Squarize, Rogerio M Castilho
PURPOSE: Emerging evidence indicates that bromodomains comprise a conserved class of epigenome readers involved in cancer development and inflammation. Bromodomains are associated with epigenetic modifications of gene transcription through interactions with lysine residues of histone tails. Particularly, the bromodomain and extra-terminal domain (BET) family member BRD4 has been found to be involved in the control over oncogenes, including c-MYC, and in the maintenance of downstream inflammatory processes...
December 11, 2018: Cellular Oncology (Dordrecht)
Weichao Zhao, Nan Hu, Dexin Ding, Dingxin Long, Sheng Li, Guangyue Li, Hui Zhang
In this paper, the developmental toxicity and apoptosis in zebrafish (Danio rerio) embryos induced by 0.01, 0.05, and 0.10-Gy γ-ray irradiation were investigated and verified by single cell gel electrophoresis, acridine orange staining, flow cytometry, transmission electron microscopy, digital gene expression sequencing, and Western blot analysis. DNA damage, deformity rates, and apoptosis of zebrafish embryos were found to increase significantly with the increase of irradiation dose, and survival and hatching rates significantly decreased when the irradiation dose exceeds 0...
December 12, 2018: Environmental Science and Pollution Research International
Joydeep Ghosh, Safa F Mohamad, Edward F Srour
Hematopoietic stem cells (HSCs) are regulated by multiple components of the hematopoietic niche, including bone marrow-derived macrophages and osteomacs. However, both macrophages and osteomacs are phenotypically similar. Thus, specific phenotypic markers are required to differentially identify the effects of osteomacs and bone marrow macrophages on different physiological processes, including hematopoiesis and bone remodeling. Here, we describe a protocol for isolation of murine bone marrow-derived macrophages and osteomacs from neonatal and adult mice and subsequent identification by multi-parametric flow cytometry using an 8-color antibody panel...
December 12, 2018: Methods in Molecular Biology
Jun-Feng Zhu, Ying Xu, Jian Zhao, Xue Li, Xinrui Meng, Tian-Qi Wang, Ben-Yao Zou, Peng-Yan Zhao, Qi Liu, Chang-Long Lu, Fang-Liang Zheng, Hong-Sheng Liu
Background: Previously, we have reported that IL-33 functioned as a protective modulator in dextran sulfate sodium- (DSS-) induced chronic colitis by suppressing Th17 cell response in colon lamina propria and IL-33 induced both regulatory B cells (Bregs) and regulatory T cells (Tregs) in mesenteric lymph nodes (MLNs) of mice with DSS-induced acute colitis. Moreover, we speculated that IL-33 would promote the Treg or Breg responses leading to the attenuation of DSS-induced chronic colitis...
2018: Journal of Immunology Research
Kerry A Casey, Xiang Guo, Michael A Smith, Shiliang Wang, Dominic Sinibaldi, Miguel A Sanjuan, Liangwei Wang, Gabor G Illei, Wendy I White
Objective: Anifrolumab is a fully human immunoglobulin G1 κ monoclonal antibody specific for subunit 1 of the type I interferon (IFN) α receptor. In a phase IIb study of adults with moderate to severe SLE, anifrolumab treatment demonstrated substantial reductions in multiple clinical endpoints. Here, we evaluated serum proteins and immune cells associated with SLE pathogenesis, type I interferon gene signature (IFNGS) test status and disease activity, and how anifrolumab affected these components...
2018: Lupus Science & Medicine
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