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Intracellular Flow Cytometry

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https://www.readbyqxmd.com/read/28938397/regulation-of-orbital-fibrosis-and-adipogenesis-by-pathogenic-th17-cells-in-graves-orbitopathy
#1
Sijie Fang, Yazhuo Huang, Sisi Zhong, Yangyang Li, Yidan Zhang, Yinwei Li, Jing Sun, Xingtong Liu, Yang Wang, Shuo Zhang, Tianle Xu, Xiaodong Sun, Ping Gu, Dan Li, Huifang Zhou, Bin Li, Xianqun Fan
Context: Th17 cells are correlated with many human autoimmune disorders including Graves' disease, and may play key roles in the pathogenesis of Graves' orbitopathy (GO). Objective: To study the phenotype of Th17 cells in patients with GO and healthy subjects, investigate the fibrosis and adipogenesis in orbital fibroblasts (OFs) modulated by IL-17A, and determine the interaction between Th17 cells and OFs. Design/Setting/Participants: Blood samples and orbital tissues from GO patients and healthy controls were collected...
August 16, 2017: Journal of Clinical Endocrinology and Metabolism
https://www.readbyqxmd.com/read/28938010/epidermal-growth-factor-receptor-targeted-sonoporation-with-microbubbles-enhances-therapeutic-efficacy-in-a-squamous-cell-carcinoma-model
#2
Fumika Hirabayashi, Kenjiro Iwanaga, Toshinori Okinaga, Osamu Takahashi, Wataru Ariyoshi, Ryo Suzuki, Mutsumi Sugii, Kazuo Maruyama, Kazuhiro Tominaga, Tatsuji Nishihara
Sonoporation is a drug and gene delivery system using ultrasonication that allows the intracellular delivery of foreign molecules that cannot enter cells under normal conditions. We previously reported that sonoporation with microbubbles (MBs) could achieve effective intracellular drug delivery to human gingival squamous carcinoma Ca9-22 cells. In this study, we developed anti-epidermal growth factor receptor (EGFR) antibody-conjugated MBs (EGFR-MBs) and evaluated their capacity to enhance anti-cancer drug toxicity in vitro and in vivo...
2017: PloS One
https://www.readbyqxmd.com/read/28933182/synthesis-and-cellular-characterization-of-various-nano-assemblies-of-cell-penetrating-peptide-epirubicin-polyglutamate-conjugates-for-the-enhancement-of-antitumor-activity
#3
Samaneh Mohammadi, Parvin Zakeri-Milani, Nasim Golkar, Samad Mussa Farkhani, Ali Shirani, Javid Shahbazi Mojarrad, Ali Nokhodchi, Hadi Valizadeh
A new class of cell penetrating peptides (CPPs) named peptide amphiphile was designed to improve the intracellular uptake and the antitumor activity of epirubicin (EPR). Various amphiphilic CPPs were synthesized by solid phase peptide synthesis method and were chemically conjugated to EPR. Their corresponding nanoparticles (CPPs-E4 and CPPs-E8) were prepared via non-covalent binding of the peptides and polyanions. Cytotoxicity and anti-proliferative activity were evaluated by MTT assay. Cellular uptake was examined by flow cytometry and fluorescence microscopy...
September 21, 2017: Artificial Cells, Nanomedicine, and Biotechnology
https://www.readbyqxmd.com/read/28928844/inhibitory-effects-of-low-intensity-pulsed-ultrasound-sonication-on-the-proliferation-of-osteosarcoma-cells
#4
Toshihiro Matsuo, Keiji Sato, Takuya Matsui, Shigeyuki Sawada, Yoshitaka Muramatsu, Katsuhisa Kawanami, Masataka Deie
To date, there is limited data on the biological effects of low-intensity pulsed ultrasound (LIPUS) on primary malignant bone tumors. The purpose of the present study was to investigate the antitumor effects of LIPUS on osteosarcoma cells. The effects of LIPUS on cell viability, induction of apoptosis, mitochondrial membrane potential and intracellular signaling molecules in the LM8 osteosarcoma cell line were investigated. LIPUS inhibited cell viability (P=0.0022) and mitochondrial membrane potential (P=0...
September 2017: Oncology Letters
https://www.readbyqxmd.com/read/28927728/cryopreservation-of-human-whole-blood-allows-immunophenotyping-by-flow-cytometry-up-to-30days-after-cell-isolation
#5
R Madelaine Paredes, Douglas K Tadaki, Amanda Sooter, Fabia Gamboni, C D R Forest Sheppard
Immunophenotyping of whole blood (WB) by flow cytometry (FC) is used clinically to assess a patient's immune status and also in biomedical research. Current protocols recommend storage of immunolabeled samples at 4°C with FC analysis to be completed within seven days. This data acquisition window can be extended to up to one year post-labeling, but this requires cryopreservation of the samples at ultra-low temperatures (≤-80°C or in liquid nitrogen). In this study we optimized a standardized cryopreservation protocol to enable preservation of immunolabeled, human WB samples at -20°C for FC and tested its effectiveness after 0, 5, 15 or 30days...
September 16, 2017: Journal of Immunological Methods
https://www.readbyqxmd.com/read/28926641/impact-of-monotherapy-on-hiv-1-reservoir-immune-activation-and-co-infection-with-epstein-barr-virus
#6
Maria Raffaella Petrara, Anna Maria Cattelan, Lolita Sasset, Riccardo Freguja, Francesco Carmona, Silvia Sanavia, Marisa Zanchetta, Paola Del Bianco, Anita De Rossi
OBJECTIVES: Although monotherapy (mART) effectiveness in maintaining viral suppression and CD4 cell count has been extensively examined in HIV-1-infected patients, its impact on HIV-1 reservoir, immune activation, microbial translocation and co-infection with Epstein-Barr Virus (EBV) is unclear. METHODS: This retrospective study involved 32 patients who switched to mART; patients were studied at baseline, 48 and 96 weeks after mART initiation. Thirty-two patients who continued combined antiretroviral therapy (cART) over the same period of time were included in the study...
2017: PloS One
https://www.readbyqxmd.com/read/28926522/bos-is-associated-with-increased-cytotoxic-proinflammatory-cd8-t-nkt-like-and-nk-cells-in-the-small-airways
#7
Greg Hodge, Sandra Hodge, Aeneas Yeo, Phan Nguyen, Emily Hopkins, Chien-Li Holmes-Liew, Paul N Reynolds, Mark Holmes
BACKGROUND: Immunosuppression therapy after lung transplantation fails to prevent bronchiolitis obliterans syndrome (BOS) in many patients, primarily a disease of the small airways. We have reported that BOS is associated with a lack of suppression of cytotoxic mediators, and proinflammatory cytokines, in peripheral blood T, NKT-like (particularly CD8+) and NK cells. We also showed a loss of glucocorticoid receptor (GCR) in proinflammatory lymphocytes after transplant. It is unknown whether these proinflammatory lymphocytes target the small and/or large airways in BOS...
October 2017: Transplantation
https://www.readbyqxmd.com/read/28919588/poly-i-c-delivery-into-j774-1-and-raw264-7-macrophages-induces-rapid-cell-death
#8
Cheah Wen Yapp, Amal Widaad Mohaimin, Adi Idris
BACKGROUND: Cytosolic double-stranded RNA (dsRNA) is an important 'molecular signature' for the detection of intracellular viral infections. Although intracellular dsRNA is a known potent inducer of apoptosis, the optimal time and dose for the onset of dsRNA-mediated apoptosis have not been studied in detail. OBJECTIVE: To perform an accurate temporal assessment of the cell death responses in dsRNA-dependent cytotoxicity. METHODS: Poly I:C (PIC), a synthetic dsRNA molecule was delivered intracellularly into J774...
September 2017: Iranian Journal of Immunology: IJI
https://www.readbyqxmd.com/read/28917946/cellular-uptake-and-intracellular-localization-of-poly-acrylic-acid-nanoparticles-in-a-rainbow-trout-oncorhynchus-mykiss-gill-epithelial-cell-line-rtgill-w1
#9
Lindsey C Felix, Van A Ortega, Greg G Goss
The ever-growing production of engineered nanoparticles (NPs) for use in many agricultural, commercial, consumer, and industrial applications will lead to their accidental or intentional release into the environment. Potential routes of environmental exposure include manufacturing or transport spills, disposal of NP-containing products down the drain and/or in landfills, as well as direct usage on agricultural land. Therefore, NPs will inevitably contaminate aquatic environments and interact with resident organisms...
September 7, 2017: Aquatic Toxicology
https://www.readbyqxmd.com/read/28916337/angiotensin-ii-induces-calcium-mediated-autophagy-in-podocytes-through-enhancing-reactive-oxygen-species-levels
#10
Na Gao, Hui Wang, Hongqiang Yin, Zhuo Yang
As well known, abnormalities of Angiotensin II (Ang II) is closely related with glomerular damage. This study was to investigate whether Ang II could affect autophagy in podocytes via oxidative stress, and whether autophagy had a positive role in protecting podocytes impaired by Ang II. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay showed that Ang II induced podocyte death. The measurements of malondialdehyde (MDA) and H2O2 levels, and flow cytometry assay revealed that Ang II considerably increased ROS generation in podocytes...
September 12, 2017: Chemico-biological Interactions
https://www.readbyqxmd.com/read/28915062/a-recombinant-fragment-of-human-surfactant-protein-d-suppresses-basophil-activation-th2-and-b-cell-responses-in-grass-pollen-induced-allergic-inflammation
#11
Asif S Qaseem, Iesha Singh, Ansar A Pathan, Janice A Layhadi, Rebecca Parkin, Fedina Alexandra, Stephen R Durham, Uday Kishore, Mohamed H Shamji
RATIONALE: rfhSP-D has been shown to suppress house dust mite and Aspergillus fumigatus-induced allergic inflammation in murine models. OBJECTIVES: We sought to elucidate the effect of rfhSP-D on FcεRI and CD23-mediated grass pollen induced allergic inflammatory responses. METHODS: rfhSP-D, containing homotrimeric neck and lectin domains, was expressed in Escherichia coli BL21 (λDE3) pLysS. PBMCs and sera were obtained from grass pollen allergic individuals (n=27)...
September 15, 2017: American Journal of Respiratory and Critical Care Medicine
https://www.readbyqxmd.com/read/28912882/atorvastatin-alleviates-iodinated-contrast-media-induced-cytotoxicity-in-human-proximal-renal-tubular-epithelial-cells
#12
Gai-Ling Liu, Rong Lei, Shao-Bin Duan, Mi-Mi Tang, Min Luo, Qian Xu
Contrast media (CM)-induced nephropathy (CIN) is a serious complication of intravascularly applied radiocontrast media. At present, no drugs have been approved for the prevention of CIN. The present study aimed to explore the effects and potential mechanisms of atorvastatin on iodinated CM-induced cytotoxicity in the human proximal renal tubular epithelial cells. The cytotoxic effect of iohexol (50, 100 and 200 mg I/ml) and the protective effect of atorvastatin pretreatment (1, 20 and 40 µM) were assessed...
October 2017: Experimental and Therapeutic Medicine
https://www.readbyqxmd.com/read/28910211/establishment-of-cmab-43-a-sensitive-and-specific-anti-cd133-monoclonal-antibody-for-immunohistochemistry
#13
Shunsuke Itai, Yuki Fujii, Takuro Nakamura, Yao-Wen Chang, Miyuki Yanaka, Noriko Saidoh, Saori Handa, Hiroyoshi Suzuki, Hiroyuki Harada, Shinji Yamada, Mika K Kaneko, Yukinari Kato
CD133, also known as prominin-1, was first described as a cell surface marker on early progenitor and hematopoietic stem cells. It is a five-domain transmembrane protein composed of an N-terminal extracellular tail, two small cytoplasmic loops, two large extracellular loops containing seven potential glycosylation sites, and a short C-terminal intracellular tail. CD133 has been used as a marker to identify cancer stem cells derived from primary solid tumors and as a prognostic marker of gliomas. Herein, we developed novel anti-CD133 monoclonal antibodies (mAbs) and characterized their efficacy in flow cytometry, Western blot, and immunohistochemical analyses...
September 14, 2017: Monoclonal Antibodies in Immunodiagnosis and Immunotherapy
https://www.readbyqxmd.com/read/28904555/acyclic-sesquiterpenes-from-the-fruit-pericarp-of-sapindus-saponaria-induce-ultrastructural-alterations-and-cell-death-in-leishmania-amazonensis
#14
Amanda Louzano Moreira, Débora Botura Scariot, Bruna Luíza Pelegrini, Greisiele Lorena Pessini, Tânia Ueda-Nakamura, Celso Vataru Nakamura, Izabel Cristina Piloto Ferreira
Previous studies reported antiprotozoal activities of Sapindus saponaria L. The aim of this work was the evaluation of antileishmanial activity and mechanism of action of extract and fractions of S. saponaria L. Hydroethanolic extract (EHA) obtained from fruit pericarps was fractionated using solid-phase extraction in a reversed phase, resulting in fractions enriched with saponins (SAP fraction) and acyclic sesquiterpene oligoglycosides (OGSA fraction). The activities of EHA, SAP, and OGSA were evaluated by antiproliferative assays with promastigote and intracellular amastigote forms...
2017: Evidence-based Complementary and Alternative Medicine: ECAM
https://www.readbyqxmd.com/read/28903821/age-and-sex-associated-differences-in-phenotypic-and-functional-characteristics-of-peripheral-blood-lymphocytes-in-chimpanzees-pan-troglodytes
#15
Pramod N Nehete, Elizabeth R Magden, Bharti P Nehete, Lawrence E Williams, Christian R Abee, K Jagannadha Sastry
Chimpanzees are the closest phylogenetic relatives to humans, sharing more than 98% genetic sequence identity. These genetic similarities prompted the belief that chimpanzees can serve as an ideal model for human disease conditions and vaccine development. However, in light of the recent NIH decision to phase out biomedical research in chimpanzees and retire NIH-supported chimpanzees, data from the present study will continue to provide value for the care of aged and sick chimpanzees located in zoos, sanctuaries, and primate centers...
September 1, 2017: Journal of the American Association for Laboratory Animal Science: JAALAS
https://www.readbyqxmd.com/read/28902941/intestinal-inflammatory-and-redox-responses-to-the-perioperative-administration-of-teduglutide-in-rats
#16
Beatriz Pinto da Costa, Ana Cristina Gonçalves, Ana Margarida Abrantes, Paulo Matafome, Raquel Seiça, Ana Bela Sarmento-Ribeiro, Maria Filomena Botelho, Francisco Castro-Sousa
Purpose: To investigate the inflammatory and redox responses to teduglutide on an animal model of laparotomy and intestinal anastomosis. Methods: Wistar rats (n=62) were allocated into four groups: "Ileal Resection and Anastomosis" vs. "Laparotomy", each one split into "Postoperative Teduglutide Administration" vs. "No Treatment"; and euthanized at the third or the seventh day. Ileal and blood samples were recovered at the baseline and at the euthanasia. Flow cytometry was used to study the inflammatory response (IL-1α, MCP-1, TNF-α, IFN-γ and IL-4 levels), oxidative stress (cytosolic peroxides, mitochondrial reactive species, intracellular glutathione and mitochondrial membrane potential) and cellular viability and death (annexin V/propidium iodide double staining)...
August 2017: Acta Cirúrgica Brasileira
https://www.readbyqxmd.com/read/28902888/dendrimer-based-selective-autophagy-induction-rescues-%C3%AE-f508-cftr-and-inhibits-pseudomonas-aeruginosa-infection-in-cystic-fibrosis
#17
Scott Mackenzie Brockman, Manish Bodas, David Silverberg, Ajit Sharma, Neeraj Vij
BACKGROUND: Cystic Fibrosis (CF) is a genetic disorder caused by mutation(s) in the CF-transmembrane conductance regulator (Cftr) gene. The most common mutation, ΔF508, leads to accumulation of defective-CFTR protein in aggresome-bodies. Additionally, Pseudomonas aeruginosa (Pa), a common CF pathogen, exacerbates obstructive CF lung pathology. In the present study, we aimed to develop and test a novel strategy to improve the bioavailability and potentially achieve targeted drug delivery of cysteamine, a potent autophagy-inducing drug with anti-bacterial properties, by developing a dendrimer (PAMAM-DEN)-based cysteamine analogue...
2017: PloS One
https://www.readbyqxmd.com/read/28901411/palmitate-induces-myocardial-lipotoxic-injury-via-the-endoplasmic-reticulum-stress%C3%A2-mediated-apoptosis-pathway
#18
Lu Zou, Xiaoyan Li, Nan Wu, Pengyu Jia, Chunting Liu, Dalin Jia
Increased free fatty acids in cardiomyocytes induce myocardial lipotoxic injury, but the underlying mechanisms are not well understood. Therefore, the aim of the present study was to explore the role of the endoplasmic reticulum (ER) stress‑mediated apoptosis pathway in palmitate (PA)‑induced cardiomyocyte lipotoxicity. H9c2 cells were treated with various doses (100, 200 and 400 µM) of PA to mimic cardiomyocyte lipotoxicity in vitro. Oil Red O staining was used to determine the accumulation of intracellular lipids...
August 31, 2017: Molecular Medicine Reports
https://www.readbyqxmd.com/read/28900849/neuroprotective-effects-of-kinetin-against-glutamate-induced-oxidative-cytotoxicity-in-ht22-cells-involvement-of-nrf2-and-heme-oxygenase-1
#19
Yunpeng Wei, Dan Liu, Yin Zheng, Chaoshuang Hao, Honglian Li, Wuqing Ouyang
Oxidative stress is considered as one of key factors related to Alzheimer's disease (AD), while kinetin (KT) has been reported to exert anti-oxidative activities as well as neuroprotective effects both in vivo and in vitro. Thus, in this study, the neuroprotective effects of KT against glutamate-induced oxidative toxicity in HT22 cells were investigated. To evaluate the anti-oxidative capabilities of KT itself, several anti-oxidative assays in vitro were conducted. To evaluate the neuroprotective effects of KT, the levels of intracellular reactive oxygen species (ROS) and calcium influx, mitochondrial membrane potential (MMP), and cell death were measured by flow cytometry...
September 12, 2017: Neurotoxicity Research
https://www.readbyqxmd.com/read/28899983/regulatory-nk-cells-mediated-between-immunosuppressive-monocytes-and-dysfunctional-t-cells-in-chronic-hbv-infection
#20
Haijun Li, Naicui Zhai, Zhongfeng Wang, Hongxiao Song, Yang Yang, An Cui, Tianyang Li, Guangyi Wang, Junqi Niu, Ian Nicholas Crispe, Lishan Su, Zhengkun Tu
BACKGROUND AND AIMS: HBV infection represents a major health problem worldwide, but the immunological mechanisms by which HBV causes chronic persistent infection remain only partly understood. Recently, cell subsets with suppressive features have been recognised among monocytes and natural killer (NK) cells. Here we examine the effects of HBV on monocytes and NK cells. METHODS: Monocytes and NK cells derived from chronic HBV-infected patients and healthy controls were purified and characterised for phenotype, gene expression and cytokines secretion by flow cytometry, quantitative real-time (qRT)-PCR, ELISA and western blotting...
September 12, 2017: Gut
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