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Flow cytometry—hematology

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https://www.readbyqxmd.com/read/29792916/flow-cytometry-based-method-facilitates-optimization-of-pma-treatment-condition-for-pma-qpcr-method
#1
Zhiqing Huang, Jianwei Zheng, Chunmei Shi, Qiang Chen
Coupling propidium monoazide (PMA) with quantitative PCR (PMA-qPCR) has been successfully applied to specific detection and quantification of viable cells in various samples. The optimal PMA treatment condition is usually determined through qPCR. However, it is a tedious, time consuming and costly process including DNA extraction and qPCR. To overcome this problem, a flow cytometry-based (FCM-based) method was first proposed in this study to replace qPCR for screening of the optimal PMA treatment condition for Helicobacter pylori, since the pure culture treated with PMA was actually a single cell suspension with fluorescent dye...
May 21, 2018: Molecular and Cellular Probes
https://www.readbyqxmd.com/read/29792857/tgf-%C3%AE-plays-a-vital-role-in-triple-negative-breast-cancer-tnbc-drug-resistance-through-regulating-stemness-emt-and-apoptosis
#2
Xiaodan Xu, Lu Zhang, Xiaogang He, Ping Zhang, Caihong Sun, Xiaojun Xu, Yaojuan Lu, Feifei Li
Triple negative breast cancer (TNBC) is the most malignant subtype of breast cancer in which the cell surface lacks usual targets for drug to exhibit its effects. Epirubicin (Epi) is widely used for TNBC, but a substantial number of patients develop Epi resistance that is usually associated with poor prognosis. Transforming growth factor (TGF-β) is a multifunctional cytokine. In recent study, it appears that TGF-β influences the cancer stem cell population, thus, the drug resistance of cancer may also be affected...
May 21, 2018: Biochemical and Biophysical Research Communications
https://www.readbyqxmd.com/read/29792428/structures-and-activities-of-tiahuramides-a-c-cyclic-depsipeptides-from-a-tahitian-collection-of-the-marine-cyanobacterium-lyngbya-majuscula
#3
Annabel Levert, Rebeca Alvariño, Louis Bornancin, Eliane Abou Mansour, Adam M Burja, Anne-Marie Genevière, Isabelle Bonnard, Eva Alonso, Luis Botana, Bernard Banaigs
The structures of three new cyclic depsipeptides, tiahuramides A (1), B (2), and C (3), from a French Polynesian collection of the marine cyanobacterium Lyngbya majuscula are described. The planar structures of these compounds were established by a combination of mass spectrometry and 1D and 2D NMR experiments. Absolute configurations of natural and nonproteinogenic amino acids were determined through a combination of acid hydrolysis, derivitization with Marfey's reagent, and HPLC. The absolute configuration of hydroxy acids was confirmed by Mosher's method...
May 24, 2018: Journal of Natural Products
https://www.readbyqxmd.com/read/29792274/ex-vivo-expanded-natural-regulatory-t-cells-from-patients-with-end-stage-renal-disease-or-kidney-transplantation-are-useful-for-autologous-cell-therapy
#4
Sybille Landwehr-Kenzel, Anne Zobel, Henrike Hoffmann, Niels Landwehr, Michael Schmueck-Henneresse, Thomas Schachtner, Andy Roemhild, Petra Reinke
Novel concepts employing autologous, ex vivo expanded natural regulatory T cells (nTreg) for adoptive transfer has potential to prevent organ rejection after kidney transplantation. However, the impact of dialysis and maintenance immunosuppression on the nTreg phenotype and peripheral survival is not well understood, but essential when assessing patient eligibility. The current study investigates regulatory T-cells in dialysis and kidney transplanted patients and the feasibility of generating a clinically useful nTreg product from these patients...
June 2018: Kidney International
https://www.readbyqxmd.com/read/29792199/nanoparticle-delivery-of-grape-seed-derived-proanthocyanidins-to-airway-epithelial-cells-dampens-oxidative-stress-and-inflammation
#5
S Castellani, A Trapani, A Spagnoletta, L di Toma, T Magrone, S Di Gioia, D Mandracchia, G Trapani, E Jirillo, M Conese
BACKGROUND: Chronic respiratory diseases, whose one of the hallmarks is oxidative stress, are still incurable and need novel therapeutic tools and pharmaceutical agents. The phenolic compounds contained in grape are endowed with well-recognized anti-oxidant, anti-inflammatory, anti-cancer, and anti-aging activities. Considering that natural anti-oxidants, such as proanthocyanidins, have poor water solubility and oral bioavailability, we have developed a drug delivery system based on solid lipid nanoparticles (SLN), apt to encapsulate grape seed extract (GSE), containing proanthocyanidins...
May 23, 2018: Journal of Translational Medicine
https://www.readbyqxmd.com/read/29792190/novel-insights-into-neuroinflammation-bacterial-lipopolysaccharide-tumor-necrosis-factor-%C3%AE-and-ureaplasma-species-differentially-modulate-atypical-chemokine-receptor-3-responses-in-human-brain-microvascular-endothelial-cells
#6
Christine Silwedel, Christian P Speer, Axel Haarmann, Markus Fehrholz, Heike Claus, Mathias Buttmann, Kirsten Glaser
BACKGROUND: Atypical chemokine receptor 3 (ACKR3, synonym CXCR7) is increasingly considered relevant in neuroinflammatory conditions, in which its upregulation contributes to compromised endothelial barrier function and may ultimately allow inflammatory brain injury. While an impact of ACKR3 has been recognized in several neurological autoimmune diseases, neuroinflammation may also result from infectious agents, including Ureaplasma species (spp.). Although commonly regarded as commensals of the adult urogenital tract, Ureaplasma spp...
May 23, 2018: Journal of Neuroinflammation
https://www.readbyqxmd.com/read/29791802/the-power-of-shielding-low-toxicity-and-high-transfection-performance-of-cationic-graft-copolymers-containing-poly-2-oxazoline-side-chains
#7
Anne-Kristin Trützschler, Tanja Bus, Martin Sahn, Anja Traeger, Christine Weber, Ulrich S Schubert
In this study we show the potential of oligo(2-ethyl-2-oxazoline) (OEtOx) shielded graft copolymers of (2 aminoethyl)-methacrylate (AEMA) and N methyl (2-aminoethyl) methacrylate (MAEMA) for pDNA delivery in HEK cells. Copolymers were synthesized via the macromonomer method using a combination of cationic ring opening polymerization (CROP) and reversible addition-fragmentation chain transfer (RAFT) polymerization to vary the degree of grafting (DG = 10% and 30%) as well as the side chain degree of polymerization (DP = 5 and 20)...
May 23, 2018: Biomacromolecules
https://www.readbyqxmd.com/read/29791595/microrna-3129-promotes-cell-proliferation-in-gastric-cancer-cell-line-sgc7901-via-positive-regulation-of-prb
#8
Shaofeng Yang, Nan Sheng, Lili Pan, Jing Cao, Jiao Liu, Ran Ma
Several microRNAs (miRNAs) have been reported as oncogenes or tumor suppressors in many cancers, including gastric cancer (GC). However, the role and molecular mechanism of miR-3129 in GC is largely unknown. We aimed to explore the function and the underlying molecular mechanism of miR-3129 in GC. Cancer tissues and corresponding adjacent tissues were collected from 50 patients with GC, and the expression of miR-3129 was detected by RT-qPCR. The expression of miR-3129 and pRb in human GC cell line SCG7091 was altered by transient transfection...
2018: Brazilian Journal of Medical and Biological Research, Revista Brasileira de Pesquisas Médicas e Biológicas
https://www.readbyqxmd.com/read/29791566/cytidine-phosphate-guanosine-oligodeoxynucleotides-in-combination-with-cd40-ligand-decrease-periodontal-inflammation-and-alveolar-bone-loss-in-a-tlr9-independent-manner
#9
Qian Zhao, Yang Hu, Shu Deng, Pei Yu, Bowen Chen, Zuomin Wang, Xiaozhe Han
Local administration of toll-like receptor 9 (TLR9), agonist cytidine-phosphate-guanosine oligodeoxynucleotide (CpG ODNs), and CD40 ligand (CD40L) can decrease ligature-induced periodontal inflammation and bone loss in wild type (WT) mouse. OBJECTIVE: This study aimed to explore whether such effect is dependent on TLR9 signaling. MATERIAL AND METHODS: Purified spleen B cells isolated from WT C57BL/6J mice and TLR9 knockout (KO) mice were cultured for 48 hours under the following conditions: CD40L, CpG+CD40L, CpG at low, medium and high doses...
2018: Journal of Applied Oral Science: Revista FOB
https://www.readbyqxmd.com/read/29790972/isolation-and-biological-characterization-of-mesenchymal-stem-cells-from-goose-dermis
#10
Jingjing Wang, Xulun Wu, Yanjie Zheng, Hebao Wen, Hongda Ji, Yuhua Zhao, Weijun Guan
The skin is a natural target of stem cell research because of its large size and easy accessibility. Cutaneous mesenchymal stem cells have shown to be a promising source of various adult stem cell or progenitor cell populations, which provide an important source of stem cell-based investigation. Nowadays, much work has been done on dermal-derived mesenchymal stem cells (DMSCs) from humans, mice, sheep, and other mammals, but the literature on avian species has been rarely reported. As an animal model, the goose is an endemic species abounding in dermal tissues which is important in the global economy...
May 21, 2018: Poultry Science
https://www.readbyqxmd.com/read/29790701/the-clinical-importance-of-changes-in-treg-and-th17-lymphocyte-subsets-in-splenectomized-patients-after-spleen-injury
#11
Witold Zgodziński, Ewelina Grywalska, Dorota Siwicka-Gieroba, Agata Surdacka, Krzysztof Zinkiewicz, Dariusz Szczepanek, Grzegorz Wallner, Jacek Roliński
BACKGROUND: Splenectomized patients are more prone to bacterial infections due to their immunocompromised status. Little is known about the role of T helper 17 (Th17) and T regulatory cells (Treg) in the immune system of patients after the removal of the spleen. OBJECTIVES: The aim of the present study was to analyze possible changes in CD4+ lymphocyte T subsets, especially Treg and Th17, in patients who had undergone splenectomy. MATERIAL AND METHODS: The study included a group of 67 male patients (41...
May 22, 2018: Advances in Clinical and Experimental Medicine: Official Organ Wroclaw Medical University
https://www.readbyqxmd.com/read/29790668/the-effects-of-aberrant-expression-of-lncrna-dgcr5-mir-873-5p-tusc3-in-lung-cancer-cell-progression
#12
Judong Luo, Hong Zhu, Hua Jiang, Yayun Cui, Mengjie Wang, Xinye Ni, Changsheng Ma
Lung cancer is the most common cause of cancer-related mortality worldwide, and nonsmall cell lung cancer (NSCLC) accounts for 80% of all pulmonary carcinomas. Recently, long noncoding RNAs (lncRNAs) have been paid attention for exploring treatment of various diseases. Upregulation of DiGeorge syndrome critical region gene 5 (DGCR5) predicts better lung squamous cell carcinoma prognosis; therefore, we explore the role of DGCR5 in lung cancer in our present study. Consecutive patients with LC were treated in our hospital between January 2015 and January 2016...
May 23, 2018: Cancer Medicine
https://www.readbyqxmd.com/read/29790655/maximising-yield-of-peripheral-blood-flow-cytometry-for-chronic-lymphoproliferative-disorders
#13
H Y Lim, F S Hong
INTRODUCTION: Flow cytometry is used in the diagnosis of haematological diseases including chronic lymphoproliferative disorders. This audit aims to ascertain the real-world indications for peripheral blood (PB) flow cytometry and which of these are associated with higher diagnostic yields. METHODS: All PB flow cytometry requests for chronic lymphoproliferative disorders from 1 January 2014 to 31 December 2014 were identified using the laboratory information system...
May 23, 2018: International Journal of Laboratory Hematology
https://www.readbyqxmd.com/read/29790576/memory-t-cell-subsets-in-healthy-gingiva-and-periodontitis-tissues
#14
Rangsini Mahanonda, Chantrakorn Champaiboon, Keskanya Subbalekha, Noppadol Sa-Ard-Iam, Arsarn Yongyuth, Benjarat Isaraphithakkul, Pimprapa Rerkyen, Orawan Charatkulangkun, Sathit Pichyangkul
BACKGROUND: In the gingival sulcus, effective and balanced innate and adaptive immune responses against subgingival plaque microbiome are crucial to maintain immune homeostasis. In this study, we investigated the memory T cell subsets in healthy gingiva and periodontitis tissues. METHODS: Anatomical localization of T cells (CD3+ , CD4+ , and CD8+ ) in healthy gingiva and periodontitis tissues were examined immunohistochemically. Subsets of memory T cells from isolated gingival cells were analyzed by flow cytometry using a cocktail of monoclonal antibodies (anti-CD69, anti-CD103, anti-CD45RA, anti-CCR7, anti-CD28, and anti-CD95)...
May 23, 2018: Journal of Periodontology
https://www.readbyqxmd.com/read/29790424/the-fate-of-autologous-endometrial-mesenchymal-stromal-cells-after-application-in-the-healthy-equine-uterus
#15
Elisabeth Rink, Teresa Beyer, Hilari French, Elaine Watson, Christine Aurich, Xavier Donadeu
Because of their distinct differentiation, immunomodulatory and migratory capacities, endometrial mesenchymal stromal cells (MSCs) may provide an optimum source of therapeutic cells not only in relation to the uterus but also for regeneration of other tissues. This study reports the fate of endometrial MSCs following intrauterine application in mares. Stromal cell fractions were isolated from endometrial biopsies taken from seven reproductively healthy mares, expanded and fluorescence-labeled in culture. MSCs (15 x 106) or PBS were autologously infused into each uterine horn during early diestrus and subsequently tracked by fluorescence microscopy and flow cytometry of endometrial biopsies and blood samples taken periodically after infusion...
May 23, 2018: Stem Cells and Development
https://www.readbyqxmd.com/read/29790256/platelet-characteristics-in-patients-with-essential-thrombocytosis
#16
Oliver Heidmann Pedersen, Mads Lamm Larsen, Erik Lerkevang Grove, Peter Buur van Kooten Niekerk, Søren Bønløkke, Peter H Nissen, Steen Dalby Kristensen, Anne-Mette Hvas
BACKGROUND: Essential thrombocytosis (ET) is a myeloproliferative disorder characterized by an increased platelet count. ET is associated with an increased risk of thrombosis, and procoagulant features of the disease may include an increased number of reactive reticulated platelets and an increased aggregation potential. We aimed to explore the association between platelet count, platelet turnover and platelet aggregation in patients with ET. METHODS: We included 24 ET patients who discontinued antiplatelet therapy prior to blood sampling...
May 23, 2018: Cytometry. Part B, Clinical Cytometry
https://www.readbyqxmd.com/read/29790189/the-identification-of-a-novel-antibody-for-cd133-using-human-antibody-phage-display
#17
Paige M Glumac, Colleen L Forster, Hong Zhou, Paari Murugan, Shilpa Gupta, Aaron M LeBeau
BACKGROUND: The transmembrane glycoprotein CD133 is believed to be a marker of adult prostate stem cells and cancer stem/initiating cells. Investigating the role of CD133 in the normal biology of the prostate and in cancer is complicated by the lack of a sensitive and accurate antibody for its detection. Here, we describe the characterization of a unique antibody identified using human antibody phage display that can recognize CD133 in both formalin-fixed tissues and cell lines. METHODS: A human single-chain variable fragment (scFv) antibody phage display library possessing a diversity of 8 × 109 was screened against fully glycosylated recombinant CD133...
May 22, 2018: Prostate
https://www.readbyqxmd.com/read/29790165/igg1-b-cell-immunity-predates-ige-responses-in-epicutaneous-sensitization-to-foods
#18
Rodrigo Jiménez-Saiz, Yosef Ellenbogen, Joshua Koenig, Melissa E Gordon, Tina D Walker, Domenico Rosace, Paul Spill, Kelly Bruton, Joshua Kong, Kristin Monteiro, Jianping Wen, Elaine I Tuomanen, Roland Kolbeck, Derek K Chu, Susan Waserman, Manel Jordana
BACKGROUND: The generation of IgE-mediated food allergy in humans is silent, and only diagnosed upon manifestation of clinical symptoms. While experimental models have been used to investigate some mechanisms of allergic sensitization, the generation of humoral immunity and memory remain to be elucidated. Here, we defined the evolution of allergen-specific B cell responses during epicutaneous sensitization to foods. METHODS: Wild-type and genetic knockout animals, and drug or antibody strategies for cell depletion and immunoglobulin signaling blockade were used to investigate epicutaneous sensitization and disease progression; we analyzed allergen-specific germinal centers and IgG1+ memory B cells by flow cytometry, evaluated humoral responses and determined clinical reactivity (anaphylaxis)...
May 22, 2018: Allergy
https://www.readbyqxmd.com/read/29790111/potential-of-the-dual-mtor-kinase-inhibitor-azd2014-to-overcome-paclitaxel-resistance-in-anaplastic-thyroid-carcinoma
#19
Zorica Milošević, Jasna Banković, Jelena Dinić, Chrisiida Tsimplouli, Evangelia Sereti, Miodrag Dragoj, Verica Paunović, Zorka Milovanović, Marija Stepanović, Nikola Tanić, Kostantinos Dimas, Milica Pešić
PURPOSE: Anaplastic thyroid carcinoma (ATC) is an aggressive, chemo-resistant malignancy. Chemo-resistance is often associated with changes in activity of the RAS/MAPK/ERK and PI3K/AKT/mTOR pathways and/or a high expression of ATP binding cassette (ABC) transporters, such as P-glycoprotein (P-gp) and breast cancer resistance protein (BCRP). To assess the therapeutic efficacy in ATC of a combination of the dual mTOR kinase inhibitor vistusertib (AZD2014) and paclitaxel (PTX), we generated a new cell line (Rho-) via the selection of human thyroid carcinoma 8505C cells that exhibit a low accumulation of rhodamine 123, which serves as a P-gp and BCRP substrate...
May 22, 2018: Cellular Oncology (Dordrecht)
https://www.readbyqxmd.com/read/29789650/use-of-dna-flow-cytometry-in-the-diagnosis-risk-stratification-and-management-of-gastric-epithelial-dysplasia
#20
Kwun Wah Wen, Peter S Rabinovitch, Danning Huang, Aras N Mattis, Gregory Y Lauwers, Won-Tak Choi
The natural history of gastric epithelial dysplasia and the consequential surveillance strategies are not well defined in the West. To date, the diagnosis relies on morphology, and no reliable adjunct methods, either immunohistochemical or molecular, have reproducibly been able to confirm the diagnosis and/or risk stratify gastric epithelial dysplasia. Yet, such a tool would be useful in confirming the diagnosis, and developing objective and rational surveillance guidelines. DNA flow cytometry was performed using formalin-fixed paraffin-embedded gastric tissue from 23 cases of high-grade dysplasia and 38 cases of low-grade dysplasia...
May 22, 2018: Modern Pathology: An Official Journal of the United States and Canadian Academy of Pathology, Inc
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