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cytometri of flow

Yao Jing, Lin Zhang, Zhiming Xu, Hao Chen, Shiming Ju, Jun Ding, Yan Guo, Hengli Tian
BACKGROUND The phosphatase actin regulator-1 (PHACTR-1) gene on chromosome 6 encodes an actin and protein phosphatase 1 (PP1) binding protein, Phactr-1, which is highly expressed in brain tissues. Phactr-1 expression is involved in physiological and pathological cerebral microvascular events. This study aimed to investigate the role of expression of Phactr-1 in a mouse brain capillary endothelial cell line, bEnd.3, by knockdown the PHACTR-1 gene. MATERIAL AND METHODS Three bEnd.3 cell groups were studied, CON (normal control cells), NC (control scramble transfected cells), and KD (cells with PHACTR-1 gene knockdown)...
February 17, 2019: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
Pengcheng Liu, Ruili Du, Xin Yu
BACKGROUND Ursolic acid is an important bioactive triterpenoid that has been reported to be of tremendous pharmacological importance. However, the anticancer potential of ursolic acid has not been examined against metastatic melanoma cells. Therefore, in this study we examined the anticancer potential of ursolic acid and its mode of action. MATERIAL AND METHODS WST-1 and colony formation assays were used for cell viability assessment. Cell cycle analysis was performed by flow cytometry. Apoptosis was detected by AO/EB staining using fluorescence microscopy...
February 17, 2019: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
Anne Conrad, Mathilde Boccard, Florent Valour, Vincent Alcazer, Aydee-Tamara Tovar Sanchez, Christian Chidiac, Frédéric Laurent, Philippe Vanhems, Gilles Salles, Karen Brengel-Pesce, Boris Meunier, Sophie Trouillet-Assant, Florence Ader
INTRODUCTION: Immune reconstitution after haematopoietic stem cell transplantation (HSCT) is a complex and dynamic process, varying from a state of nearly complete immunosuppression to an expected full immune recovery. Specific vaccination guidelines recommend reimmunisation after HSCT but data regarding vaccine efficacy in this unique population are scarce. New immune functional assays could enable prediction of vaccine response in the setting of HSCT. METHODS AND ANALYSIS: A prospective, longitudinal single-centre cohort study of autologous and allogeneic HSCT recipients was designed in order to determine the vaccine response to five vaccine targets (pneumococcus, hepatitis B virus, Haemophilus Influenzae type b, tetanus and diphtheria) and to correlate it to immune function parameters...
February 15, 2019: BMJ Open
Cheng Jiayi, Ning Tianyi, Teng Dan, Kang Tingguo, Wang Qingfeng, Zhang Qianqian
To explore the protective effect and mechanism of isorhamnetin against oxidative injury caused by H2 O2 to endothelial cell strain CRL1730 of human umbilical vein. H2 O2 and endothelial cell strain CRL1730 were used, as a model of injured endothelial cell. Three levels of crude drugs areorhamnetin, 22.8μg/ml, 11.4μg/mL and 5.7μg/mL was added to the injured cell strain CRL1730 respectively. The cell injury was measured in terms of necrotic rate, quantities of von Wilebr and factor (vWf) and thrombomodulin (TM), lactate dehydrogenase (LDH) and intracellular free calcium ions through flow cytometry, ELISA, fluorescent spectrometer and laser scanning confocal microscopy respectively...
January 2019: Pakistan Journal of Pharmaceutical Sciences
Katrine Pilely, Siril S Bakke, Yaseelan Palarasah, Mikkel-Ole Skjoedt, Emil D Bartels, Terje Espevik, Peter Garred
BACKGROUND AND AIMS: Cholesterol crystal (CC)-induced inflammation is a critical step in the development of atherosclerosis. CCs activate the complement system and induce an inflammatory response resulting in phagocytosis of the CCs, production of reactive oxygen species (ROS) and release of cytokines. The cyclodextrin 2-hydroxypropyl-β-cyclodextrin has been found to reduce CC-induced complement activation and induce regression of established atherosclerotic plaques in a mouse model of atherosclerosis, thus inhibition of complement with cyclodextrins is a potential new strategy for treatment of inflammation during atherosclerosis...
February 3, 2019: Atherosclerosis
Dominic Lenz, Christian Staufner, Selina Wächter, Maike Hagedorn, Juliane Ebersold, Gudrun Göhring, Stefan Kölker, Georg F Hoffmann, Sabine Jung-Klawitter
Fibroblasts of a patient with Infantile Liver Failure Syndrome 2 (OMIM #616483) due to a homozygous missense variant in the neuroblastoma amplified sequence gene (NBAS; c.[2708T>G]; c.[2708T>G]/p.[Leu903Arg]; p.[Leu903Arg]) were reprogrammed to iPSCs using the Cytotune®-iPS 2.0 Sendai Reprogramming Kit (Invitrogen) delivering the reprogramming factors Oct3/4, Sox2, c-Myc and Klf4. Cells showed a normal karyotype. Pluripotency of DHMCi004-A was proven using immunohistochemistry, RT-PCR analysis, flow cytometry and differentiation into all three germ layers using the STEMdiff™ Trilineage Differentiation Kit (Stemcell Technologies)...
February 11, 2019: Stem Cell Research
Laura Juárez-Chávez, Socorro Pina-Canseco, Delia Soto-Castro, Rosa Santillan, Nancy E Magaña-Vergara, Paz María Salazar-Schettino, Margarita Cabrera-Bravo, Eduardo Pérez-Campos
The increasing use of dendrimers shows promise for the treatment of inflammatory diseases, Chagas disease and other conditions such as cancer. In this study, the activity of 1st and 2nd generation dendrimers over T. cruzi in the epimastigote stage was tested. Dendrimers were derived from α-ethynylestradiol (EE) modified with PAMAM-type dendrons through a triazole ring. The activity of each compound was evaluated in five doses (from 1.3 to 20 µmol/mL) by flow cytometry, including benznidazole (Bz) as positive control...
January 28, 2019: Bioorganic Chemistry
Min Ding, Qian-Hua Fang, Yuan-Tao Cui, Qi-Ling Shen, Qian Liu, Peng-Hua Wang, De-Min Yu, Chun-Jun Li
AIMS: High glucose (HG)-induced pancreatic β-cell apoptosis may be a major contributor to the progression of diabetes mellitus (DM). NADPH oxidase (NOX2) has been considered a crucial regulator in β-cell apoptosis. This study was designed to evaluate the impact of GLP-1 receptor agonist (GLP-1Ra) liraglutide on pancreatic β-cell apoptosis in diabetes and the underlying mechanisms involved. METHODS: The diabetic rat models induced by streptozotocin (STZ) and a high fat diet (HFD) received 12 weeks of liraglutide treatment...
January 2, 2019: Journal of Diabetes and its Complications
Jessica Jane Browne, Evan Howard Matthews, Andrew William Taylor-Robinson, Jennelle Maree Kyd
OBJECTIVES: Regulatory T lymphocytes (Treg ) have been linked to survival of commensal bacteria at mucosal sites, but their presence and role in chronic otitis media (COM) and their response to otopathogens has not been evaluated previously. We investigated the association between Treg lymphocytes and otopathogens in COM prone and non-COM prone children. METHODS: Forty children, 2-7 years of age, scheduled for adenoidectomy were enrolled into COM (n = 20) or non-COM (n = 20) groups...
February 5, 2019: International Journal of Pediatric Otorhinolaryngology
Jong Woo Lee, Yu Zhang, Kyung Jin Eoh, Roshan Sharma, Miguel F Sanmamed, Jenny Wu, Justin Choi, Hee Sun Park, Akiko Iwasaki, Edward Kaftan, Lieping Chen, Vali Papadimitrakopoulou, Roy S Herbst, Ja Seok Koo
PURPOSE: To characterize the tumor-infiltrating immune cells population in Kras/p53-driven lung tumors and to evaluate the combinatorial anti-tumor effect with MEK inhibitor (MEKi), trametinib, and immunomodulatory monoclonal antibodies (mAbs) targeting either programmed cell death protein-1 (PD-1) or programmed cell death protein ligand 1 (PD-L1) in vivo. EXPERIMENTAL DESIGN: Trp53FloxFlox ;KrasG12D/+ ;Rosa26LSL-Luciferase/LSL-Luciferase (PKL) genetically engineered mice were utilized to develop autochthonous lung tumors with intratracheal delivery of adenoviral Cre recombinase...
February 13, 2019: Journal of Thoracic Oncology
Mario T Schellenberger, Udo Bock, Jennifer Hennen, Florian Groeber-Becker, Heike Walles, Brunhilde Blömeke
A key event of the adverse outcome pathway for skin sensitization is the activation of dendritic cells (DC). To be most close to the in vivo situation, we combined for the first time reconstructed human epidermis (RHE) in coculture with THP-1 cells, as surrogate for DC. THP-1 cells were placed underneath RHE (SkinEthic™, OS-REp). Cell activation was measured by analyzing cell surface expression of co-stimulatory molecules CD86 and CD40, adhesion molecule CD54 and of human leukocyte antigen class II-related subtypes (HLA-DR) on THP-1 cells by flow cytometry...
February 13, 2019: Toxicology in Vitro: An International Journal Published in Association with BIBRA
Jian Zhang, Huali Zhang, Yongli Luo
BACKGROUND The aim of this study was to investigate the clinical correlation between sPD-1 (soluble programmed cell death-1) and PD-1 (programmed cell death-1) expression and cancer pain. MATERIAL AND METHODS sPD-1 content in peripheral blood was determined by enzyme-linked immunosorbent assay (ELISA). T cell surface-positive rate was determined by flow cytometry, and the correlation of clinical characteristics of patients with cancer pain was analyzed. RESULTS The positive expression rate of PD-1 in sPD-1 and T cells of patients with cancer pain was higher than that in normal patients...
February 16, 2019: Medical Science Monitor: International Medical Journal of Experimental and Clinical Research
Huiming Wang, Changxuan Liu, Wenli Chen, Guohua Ding
CD19+ CD24hi CD38hi cells play an essential role in maintaining immune homeostasis. CD40 signaling is involved in regulating the induction and function of CD19+ CD24hi CD38hi cells. Changes in B-cell subpopulations and CD19+ CD24hi CD38hi cells have been observed in systemic lupus erythematosus (SLE) patients. Whether changes in the B-cell subpopulation are related to the aberrant CD40 signaling in SLE patients remains unclear. In this study, we examined changes in the levels of CD19+ CD24hi CD38hi cells and CD19+ CD24hi CD38low cells in peripheral blood mononuclear cells and the serum level of soluble CD40 ligand (sCD40L) in 30 patients with SLE...
February 16, 2019: Journal of Cellular Biochemistry
A Juszkiewicz, A Glapa, P Basta, E Petriczko, K Żołnowski, B Machaliński, J Trzeciak, K Łuczkowska, A Skarpańska-Stejnborn
BACKGROUND: The aim of this study was to analyze the response of selected components of the immune system in rowers to maximal physical exercise, and to verify if this response could be modulated by supplementation with L-theanine. METHOD: The double-blind study included 20 members of the Polish Rowing Team. The subjects were randomly assigned to the supplemented group (n = 10), receiving 150 mg of L-theanine extract for 6 weeks, or to the placebo group (n = 10)...
February 15, 2019: Journal of the International Society of Sports Nutrition
Mingshui Chen, Aditi Sharma, Yanling Lin, Yanheng Wu, Qi He, Yushu Gu, Zhi Ping Xu, Michael Monteiro, Wenyi Gu
BACKGROUND: Programmed cell death ligand 1 (PD-L1) is an important immune-inhibitory protein expressed on cancer cells to mediate cancer escape through interaction with PD-1 expressed on activated T lymphocytes (T cells). Previously, we reported that colon and breast cancer stem cells (CSCs) expressed much higher levels of PD-L1 than their parental cells, suggesting they will be more resistant to immune attack. METHODS: We investigated the underlining mechanism of PD-L1 increase in colon CSCs, with a special focus on the effect of insulin and epithelial growth factor (EGF), the two fundamental components to sustain the metabolism and stemness in the culture of CSCs...
February 15, 2019: BMC Cancer
Aishwarya Satish, Purna Sai Korrapati
Nerve injuries are often debilitating as its regeneration occurs in a slow and laborious manner. Remediation of nerve injury is a colossal task as functional restoration in larger gaps seldom occurs due to the complex nerve regeneration mechanism. A nanofiber-based graft material has been fabricated to provide topographical and biochemical cues to encourage neural differentiation. Laminin plays a crucial role in supporting peripheral nerve regeneration and hence aligned polyvinyl cinnamate nanofibers surface-conjugated with laminin-derived cell-adhesion peptides have been fabricated to improve selective neural adhesion and regeneration...
February 15, 2019: Journal of Tissue Engineering and Regenerative Medicine
William Wheat, Lyndah Chow, Jonathan Coy, Elena Contreras, Michael Lappin, Steven Dow
BACKGROUND: Nonspecific induction of local innate immune responses by mucosally administered immunotherapy is a new approach to protection from upper respiratory tract infections. Therefore, a new liposome-toll-like receptor complex (LTC) immune stimulant was developed and investigated for its ability to activate innate immune responses in cats, both in vitro and in vivo, as part of an initial evaluation of LTC for use as an immunotherapeutic agent in cats. OBJECTIVES: We hypothesized that LTC could activate innate immune responses in cats after topical application to nasal and oropharyngeal mucosal surfaces...
February 15, 2019: Journal of Veterinary Internal Medicine
Qi Li, Sahar Shaikh, Hayato Iwase, Cassandra Long, Whayoung Lee, Zhongqiang Zhang, Yi Wang, David Ayares, David K C Cooper, Hidetaka Hara
BACKGROUND: Old World non-human primates (OWNHPs) are used for preclinical pig-to-NHP studies. However, like pigs, OWNHPs express Neu5Gc, and therefore do not develop natural anti-Neu5Gc antibodies. New World NHPs (NWNHPs) have been reported not to express Neu5Gc. We investigated the potential of NWNHPs in xenotransplantation research. METHODS: We investigated expression of Gal, Neu5Gc, and Sda antigens on RBCs and PBMCs from humans, selected OWNHPs, and capuchin monkeys (a NWNHP)...
February 16, 2019: Xenotransplantation
Yasuhiro Shimojima, Dai Kishida, Ken-Ichi Ueno, Satoru Ushiyama, Takanori Ichikawa, Yoshiki Sekijima
OBJECTIVE: To investigate the characteristics of circulating NK cells and their IFN-γ-producing ability in adult-onset Still's disease (AOSD). METHODS: Peripheral blood mononuclear cells were obtained from 22 patients in the acute phase of AOSD (acute AOSD); 7 of the 22 patients after treatment (remission AOSD), and 11 healthy controls (HC). NK cells and their IFN-γ expression levels were analyzed by flow cytometry. Additionally, the cytokine receptors of interleukin (IL)-12, IL-15, and IL-18 on NK cells were also evaluated...
February 15, 2019: Journal of Rheumatology
Magnus T Dillon, Katharina F Bergerhoff, Malin Pedersen, Emmanuel C Patin, Harriet Whittock, Eva Crespo-Rodriguez, Alex Pearson, James T Paget, Henry G Smith, Radhika R Patel, Shane Foo, Galabina Bozhanova, Chanthirika Ragulan, Elisa Fontana, Krisha Desai, Anna C Wilkins, Anguraj Sadanandam, Alan Melcher, Martin McLaughlin, Kevin J Harrington
PURPOSE: ATR inhibitors (ATRi) are in early phase clinical trials and have been shown to sensitise to chemotherapy and radiotherapy preclinically. No data have been published about the effect of these drugs on the tumor microenvironment. EXPERIMENTAL DESIGN: We used an immunocompetent mouse model of HPV-driven malignancies to investigate the ATR inhibitor AZD6738 in combination with fractionated radiation (RT). Gene expression analysis and flow cytometry were performed post-therapy...
February 15, 2019: Clinical Cancer Research: An Official Journal of the American Association for Cancer Research
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