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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/28439677/ap-2%C3%AE-reverses-vincristine-induced-multidrug-resistance-of-sgc7901-gastric-cancer-cells-by-inhibiting-the-notch-pathway
#1
Wei Lian, Li Zhang, Long Yang, Wensheng Chen
Multidrug resistance (MDR) remains a major clinical obstacle in the treatment of gastric cancer (GC) since it causes tumor recurrence and metastasis. The transcription factor activator protein-2α (AP-2α) has been implicated in drug-resistance in breast cancer; however, its effects on MDR of gastric cancer are far from understood. In this study, we aimed to explore the effects of AP-2α on the MDR in gastric cancer cells selected by vincristine (VCR). Decreased AP-2α levels were markedly detected by RT-PCR and Western blot in gastric cancer cell lines (BGC-823, SGC-7901, AGS, MKN-45) compared with that in the gastric epithelial cell line (GES-1)...
April 24, 2017: Apoptosis: An International Journal on Programmed Cell Death
https://www.readbyqxmd.com/read/28438973/pulmonary-il-17-lymphocytes-increase-during-pneumocystis-murina-infection-but-are-not-required-for-clearance-of-pneumocystis
#2
Chiara Ripamonti, Lisa R Bishop, Joseph A Kovacs
Pneumocystis remains an important pathogen of immunosuppressed patients, causing a potentially life-threatening pneumonia. Despite its medical importance, the immune responses required to control infection, including the role of IL-17, which is important in controlling other fungal infections, have not been clearly defined. Using flow cytometry and intracellular cytokine staining following stimulation with PMA and ionomycin, we examined interferon-γ, IL-4, IL-5, and IL-17 production by lung lymphocytes in immunocompetent C57BL/6 mice over time following infection with P...
April 24, 2017: Infection and Immunity
https://www.readbyqxmd.com/read/28438610/hyperglycemia-increases-interstitial-cells-of-cajal-via-mapk1-and-mapk3-signaling-to-etv1-and-kit-leading-to-rapid-gastric-emptying
#3
Yujiro Hayashi, Yoshitaka Toyomasu, Siva Arumugam Saravanaperumal, Michael R Bardsley, John A Smestad, Andrea Lorincz, Seth T Eisenman, Gianluca Cipriani, Molly H Nelson Holte, Fatimah J Al Khazal, Sabriya A Syed, Gabriella B Gajdos, Kyoung Moo Choi, Gary J Stoltz, Katie E Miller, Michael L Kendrick, Brian P Rubin, Simon J Gibbons, Adil E Bharucha, David R Linden, Louis James Maher, Gianrico Farrugia, Tamas Ordog
BACKGROUND & AIMS: Depletion of interstitial cells of Cajal (ICCs) is common in diabetic gastroparesis. However, in approximately 20% of patients with diabetes, gastric emptying (GE) is accelerated. GE is also faster in obese individuals, and is associated with increased blood levels of glucose in patients with type 2 diabetes. To understand the fate of ICCs in hyperinsulinemic, hyperglycemic states characterized by rapid GE, we studied mice with mutation of the leptin receptor (Lepr(db/db)), which in our colony had accelerated GE...
April 21, 2017: Gastroenterology
https://www.readbyqxmd.com/read/28435822/targeted-intracellular-heat-transfer-in-cancer-therapy-assessment-of-asparagine-laminated-gold-nanoparticles-in-cell-model-of-t-cell-leukemia
#4
Shadab Shahriari, Maryam Bakhshi, Ahmad Reza Shahverdi, Azar Berahmeh, Farnaz Safavifar, Mohammad Reza Khorramizadeh
BACKGROUND: High temperatures have destructive effects on cancer cells by damaging proteins and structures within cells. Gold nanoparticles (AuNPs) can act as drug delivery vehicles, especially for cancer therapy. Due to the selective intake of asparagine molecules into malignant cells, AuNPs were coated with asparagine; and CCRF-CEM human T-cell leukemia cells were treated with the new combination, Asn-AuNPs, at 39 °C. METHODS: The co-authors from a number of collaborative labs located at Tehran University of Medical Sciences, Tehran, Iran, have initiated the idea and preliminary design of this study in 2011...
March 2017: Iranian Journal of Public Health
https://www.readbyqxmd.com/read/28434981/protein-transport-inhibitors-downregulate-the-expression-of-lag-3-on-regulatory-t-cells
#5
Sara Anvari, Andrea Grimbergen, Carla M Davis, George Makedonas
Modern immunologic studies demand increasing complexity because of a need to improve our understanding of the relationship between a cell's phenotype and its function. Regulatory T cells have been defined by a narrow set of phenotypic markers, however their actual functional capacity has not been determined at the single-cell level. Although the lymphocyte activation gene 3 (LAG-3; CD223) is a key marker for the identification of exhausted T cells, it may be useful also in resolving Treg subpopulations by indicating distinct functional breadths...
April 20, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28433940/a-novel-dual-ratiometric-response-fluorescent-probe-for-so2-clo-detection-in-cells-and-in%C3%A2-vivo-and-its-application-in-exploring-the-dichotomous-role-of-so2-under-the-clo-induced-oxidative-stress
#6
Kun Dou, Qiang Fu, Guang Chen, Fabiao Yu, Yuxia Liu, Ziping Cao, Guoliang Li, Xianen Zhao, Lian Xia, Lingxin Chen, Hua Wang, Jinmao You
Intracellular reactive sulfur species and reactive oxygen species play vital roles in immunologic mechanism. As an emerging signal transmitter, SO2 can be generated as the anti-oxidant, while SO2 is also a potential oxidative stress-inducer in organism. Aiming to elucidate in-depth the dichotomous role of SO2 under oxidative stress, we designed a dual-response fluorescent probe that enabled the respective or successive detection of SO2 and ClO(-). The probe itself emits the red fluorescence (625 nm) which can largely switch to blue (410 nm) and green fluorescence (500 nm) respectively in response to SO2 and ClO(-), allowing the highly selective and accurate ratiometric quantification for both SO2 and ClO(-) in cells...
April 14, 2017: Biomaterials
https://www.readbyqxmd.com/read/28431580/cripto-1-acts-as-a-functional-marker-of-cancer-stem-like-cells-and-predicts-prognosis-of-the-patients-in-esophageal-squamous-cell-carcinoma
#7
Qiang Liu, Xiang Cui, Xi Yu, Bai-Shi-Jiao Bian, Feng Qian, Xu-Gang Hu, Cheng-Dong Ji, Lang Yang, Yong Ren, Wei Cui, Xia Zhang, Peng Zhang, Ji Ming Wang, You-Hong Cui, Xiu-Wu Bian
BACKGROUND: Esophageal squamous cell carcinoma (ESCC) is highly malignant with highly invasive and metastatic capabilities and poor prognosis. It is believed that the ESCC cancer stem-like cells (ECSLCs) are critical for tumorigenicity, invasion and metastasis of ESCC. However, the properties of ECSLCs vary with different markers used in isolation, so that new and more effective markers of ECSLCs need to be identified. This study aimed to estimate the potentiality of Cripto-1 (CR-1) as an ECSLC surface marker and investigate the clinical significance of CR-1 expression in ESCC...
April 21, 2017: Molecular Cancer
https://www.readbyqxmd.com/read/28426994/silica-nanoparticles-induced-intrinsic-apoptosis-in-neuroblastoma-sh-sy5y-cells-via-cytc-apaf-1-pathway
#8
Yanyan Yang, Yongbo Yu, Jiahui Wang, Yanbo Li, Yang Li, Jia Wei, Tong Zheng, Minghua Jin, Zhiwei Sun
The present study was to investigate effects of Silica nanoparticles (SiNPs) on nervous system and explore potential mechanisms in human neuroblastoma cells (SH-SY5Y). Cytotoxicity was detected by cell viability and Lactate dehydrogenase (LDH) release. Flow cytometry analysis was applied to assess mitochondrial membrane potential (MMP) loss, intracellular Ca(2+) and apoptosis. To clarify the mechanism of SiNPs-induced apoptosis, intrinsic apoptosis-related proteins were detected. Our results showed that SiNPs caused cytotoxicity, cell membrane damage and Ca(2+) increase in a dose-dependent manner in SH-SY5Y cells...
January 30, 2017: Environmental Toxicology and Pharmacology
https://www.readbyqxmd.com/read/28426982/redox-environment-in-stem-and-differentiated-cells-a-quantitative-approach
#9
O G Lyublinskaya, Ju S Ivanova, N A Pugovkina, I V Kozhukharova, Z V Kovaleva, A N Shatrova, N D Aksenov, V V Zenin, Yu A Kaulin, I A Gamaley, N N Nikolsky
Stem cells are believed to maintain a specific intracellular redox status through a combination of enhanced removal capacity and limited production of ROS. In the present study, we challenge this assumption by developing a quantitative approach for the analysis of the pro- and antioxidant ability of human embryonic stem cells in comparison with their differentiated descendants, as well as adult stem and non-stem cells. Our measurements showed that embryonic stem cells are characterized by low ROS level, low rate of extracellular hydrogen peroxide removal and low threshold for peroxide-induced cytotoxicity...
April 11, 2017: Redox Biology
https://www.readbyqxmd.com/read/28425973/edim-tktl1-apo10-blood-test-an-innate-immune-system-based-liquid-biopsy-for-the-early-detection-characterization-and-targeted-treatment-of-cancer
#10
REVIEW
Johannes F Coy
Epitope detection in monocytes (EDIM) represents a liquid biopsy exploiting the innate immune system. Activated monocytes (macrophages) phagocytose unwanted cells/cell fragments from the whole body including solid tissues. As they return to the blood, macrophages can be used for a non-invasive detection of biomarkers, thereby providing high sensitivity and specificity, because the intracellular presence of biomarkers is due to an innate immune response. Flow cytometry analysis of blood enables the detection of macrophages and phagocytosed intracellular biomarkers...
April 20, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28425536/cisplatin-resistance-in-cell-models-evaluation-of-metallomic-and-biological-predictive-biomarkers-to-address-early-therapy-failure
#11
Marta Espina, Mario Corte-Rodríguez, Leticia Aguado, María Montes-Bayón, Marta I Sierra, Pablo Martínez-Camblor, Elisa Blanco-González, L María Sierra
Cisplatin, one of the most extensively used metallodrugs in cancer treatment, presents the important drawback of patient resistance. This resistance is the consequence of different processes including those preventing the formation of DNA adducts and/or their quick removal. Thus, a tool for the accurate detection and quantitation of cisplatin-induced adducts might be valuable for predicting patient resistance. To prove the validity of such an assumption, highly sensitive plasma mass spectrometry (ICP-MS) strategies were applied to determine DNA adduct levels and intracellular Pt concentrations...
April 20, 2017: Metallomics: Integrated Biometal Science
https://www.readbyqxmd.com/read/28423568/sepia-ink-oligopeptide-induces-apoptosis-and-growth-inhibition-in-human-lung-cancer-cells
#12
Zhi Zhang, Lei Sun, Guoren Zhou, Peng Xie, Jinjun Ye
Sepia ink oligopeptide (SIO), as a tripeptide extracted from Sepia ink, could be used as an inducer of apoptosis in human prostate cancer cells. We designed a cyclo-mimetic peptide of SIO by introducing a disulfide bond to stabilize the native peptide into beta turn structure, and produced a peptide with higher cell permeability and stability. Through labeling an FITC to the N-terminus of the peptide, the cell permeability was examined. Stabilized peptide showed enhanced cellular uptake than linear tripeptide as indicated by flow cytometry and cell fluorescent imaging...
April 4, 2017: Oncotarget
https://www.readbyqxmd.com/read/28422515/allergen-induced-increases-in-sputum-levels-of-group-2-innate-lymphoid-cells-in-asthmatic-subjects
#13
Ruchong Chen, Steven G Smith, Brittany Salter, Amani El-Gammal, John Paul Oliveria, Caitlin Obminski, Rick Watson, Paul M O'Byrne, Gail M Gauvreau, Roma Sehmi
RATIONALE: Group 2 innate lymphoid cells (ILC2), a major source of type 2 cytokines, initiate eosinophilic inflammatory responses in murine models of asthma. OBJECTIVES: Investigate the role of ILC2 in allergen-induced airway eosinophilic responses in atopic asthmatics. METHODS: Using a diluent-controlled allergen challenge cross-over study, where all subjects (n=10) developed allergen-induced early and late responses , airway eosinophilia and increased methacholine airway responsiveness, bone marrow, blood and sputum samples were collected pre- and post-inhalation challenge...
April 19, 2017: American Journal of Respiratory and Critical Care Medicine
https://www.readbyqxmd.com/read/28417934/expression-of-adenosine-a2b-receptor-and-adenosine-deaminase-in-rabbit-gastric-mucosa-ecl-cells
#14
Rosa María Arin, Ana Isabel Vallejo, Yuri Rueda, Olatz Fresnedo, Begoña Ochoa
Adenosine is readily available to the glandular epithelium of the stomach. Formed continuously in intracellular and extracellular locations, it is notably produced from ATP released in enteric cotransmission. Adenosine analogs modulate chloride secretion in gastric glands and activate acid secretion in isolated parietal cells through A2B adenosine receptor (A2BR) binding. A functional link between surface A2BR and adenosine deaminase (ADA) was found in parietal cells, but whether this connection is a general feature of gastric mucosa cells is unknown...
April 12, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28416745/stable-h3-peptide-was-delivered-by-gold-nanorods-to-inhibit-lsd1-activation-and-induce-human-mesenchymal-stem-cells-differentiation
#15
Xin Meng, Jianping Li, Minjuan Zheng, Lei Zuo, Chao Sun, Yongsheng Zhu, Ling Fang, Liwen Liu, Xiaodong Zhou
Recently, lysine-specific demethylase 1 (LSD1), which is the first identified histone demethylase, regulates post-translational modifications and has great promise as new targets for cancer and other diseases. Moreover, the ability of LSD1 to induce the differentiation of stem cells has attracted great attention in biological fields. In this study, we designed LSD1 peptide inhibitor based on its substrate H3 peptide. Through introducing a disulfide bond to stabilize the native peptide into alpha helical structure, we get a peptide with higher cell permeability and stability compared to its parent form...
April 4, 2017: Oncotarget
https://www.readbyqxmd.com/read/28416516/semaphorin-4d-inhibits-neutrophil-activation-and-is-involved-in-the-pathogenesis-of-neutrophil-mediated-autoimmune-vasculitis
#16
Masayuki Nishide, Satoshi Nojima, Daisuke Ito, Hyota Takamatsu, Shohei Koyama, Sujin Kang, Tetsuya Kimura, Keiko Morimoto, Takashi Hosokawa, Yoshitomo Hayama, Yuhei Kinehara, Yasuhiro Kato, Takeshi Nakatani, Yoshimitsu Nakanishi, Takeshi Tsuda, Jeong Hoon Park, Toru Hirano, Yoshihito Shima, Masashi Narazaki, Eiichi Morii, Atsushi Kumanogoh
OBJECTIVES: Inappropriate activation of neutrophils plays a pathological role in antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). The aim of this study was to investigate the functions of semaphorin 4D (SEMA4D) in regulation of neutrophil activation, and its involvement in AAV pathogenesis. METHODS: Serum levels of soluble SEMA4D were evaluated by ELISA. Blood cell-surface expression of membrane SEMA4D was evaluated by flow cytometry. To determine the functional interactions between neutrophil membrane SEMA4D and endothelial plexin B2, wild-type and SEMA4D(-/-) mice neutrophils were cultured with an endothelial cell line (MS1) stained with SYTOX green, and subjected to neutrophil extracellular trap (NET) formation assays...
April 17, 2017: Annals of the Rheumatic Diseases
https://www.readbyqxmd.com/read/28416440/inhibition-of-the-pi3k-akt-mtor-signaling-pathway-in-t-lymphocytes-in-patients-with-active-tuberculosis
#17
XueXuan Zhang, TengYi Huang, Ying Wu, WenGuang Peng, HanBin Xie, MeiChen Pan, HuanBin Zhou, BoZhi Cai, YingE Wu
OBJECTIVES: To investigate PI3K-Akt-mTOR signaling pathway changes and the proliferation of FoxP3(+)Treg cells in patients with active tuberculosis. METHODS: We isolated PBMCs and CD4(+)CD25(+)FoxP3(+) Treg cells from peripheral blood collected from patients with active tuberculosis and healthy controls. We compared the proportion and MFI of PI3K-Akt-mTOR pathway components and PTEN by flow cytometry using specific cell-surface and intracellular markers. Moreover, we detected the specific secretory proteins ESAT-6 and Ag85B, cytokines IL-10, TGF-β1 and IL-35 in serum by ELISA...
April 14, 2017: International Journal of Infectious Diseases: IJID
https://www.readbyqxmd.com/read/28415949/%C3%AE-catenin-promotes-intracellular-bacterial-killing-via-suppression-of-pseudomonas-aeruginosa-triggered-macrophage-autophagy
#18
Qiang Fu, Kang Chen, Qian Zhu, Weijia Wang, Fuda Huang, Lishao Miao, Xinger Wu
Objective To investigate β-catenin-mediated bacterial elimination during Pseudomonas aeruginosa infection of macrophage-like RAW264.7 cells. Methods Cell viability and catenin beta 1 ( CTNNB1) expression in RAW264.7 cells following P. aeruginosa infection versus uninfected cells, were detected by cell counting kit-8 assay and β-catenin Western blots. RAW264.7 cells with CTNNB1 overexpression were established with β-catenin lentivirus using flow cytometry and clonogenic limiting dilution assays. Bacterial killing was measured by plate counts; phagocytosis and nitric oxide (NO) were measured by flow cytometry; and reactive oxygen species (ROS) were measured using Griess reaction...
April 2017: Journal of International Medical Research
https://www.readbyqxmd.com/read/28415571/increased-b-cell-activation-is-present-in-jak2v617f-mutated-calr-mutated-and-triple-negative-essential-thrombocythemia
#19
Ken-Hong Lim, Caleb Gon-Shen Chen, Yu-Cheng Chang, Yi-Hao Chiang, Chen-Wei Kao, Wei-Ting Wang, Chiao-Yi Chang, Ling Huang, Ching-Sung Lin, Chun-Chia Cheng, Hung-I Cheng, Nai-Wen Su, Johnson Lin, Yi-Fang Chang, Ming-Chih Chang, Ruey-Kuen Hsieh, Huan-Chau Lin, Yuan-Yeh Kuo
Essential thrombocythemia (ET) is a BCL-ABL1-negative myeloproliferative neoplasm. We have reported that increased activated B cells can facilitate platelet production mediated by cytokines regardless JAK2 mutational status in ET. Recently, calreticulin (CALR) mutations were discovered in ~30% JAK2/MPL-unmutated ET and primary myelofibrosis. Here we sought to screen for CALR mutations and to evaluate B cell immune profiles in a cohort of adult Taiwanese ET patients. B cell populations, granulocytes/monocytes membrane-bound B cell-activating factor (mBAFF) levels, B cells toll-like receptor 4 (TLR4) expression and intracellular levels of interleukin (IL)-1β/IL-6 and the expression of CD69, CD80, and CD86 were quantified by flow cytometry...
March 18, 2017: Oncotarget
https://www.readbyqxmd.com/read/28415521/effect-of-halloysite-nanotubes-on-the-structure-and-function-of-important-multiple-blood-components
#20
Keke Wu, Ru Feng, Yanpeng Jiao, Changren Zhou
Many researchers have investigated the application of halloysite nanotubes (HNTs) in biomedicine, because of their special nanoscale hollow tubular structure. Although the cytocompatibility of HNTs has been studied, their blood compatibility has not been systematically investigated. In this work, the effect of HNTs on the structure and function of different blood components has been studied, including the morphology and hemolysis of red blood cells (RBCs). Based on scanning electron microscopy (SEM) observations, optical density test and flow cytometry analysis, we found that HNTs can affect the morphology and membrane integrity of RBCs in phosphate buffered saline (PBS) in a content-dependent way...
June 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
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