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Cell washing

Narendra Kumar, Vijay Kumar, Rohit Panwar, Chand Ram
The toxic heavy metal cadmium (Cd) appears as one of the major global threats to human and animal health. Human being and aquatic life are exposed to Cd by breathing, eating, or drinking when industrial effluents released into environment. The study was aimed to identify cadmium-binding Lactobacillus strain to reduce its bioaccessibility in in vitro digestion model. In this context, forty-eight lactobacilli strains isolated and characterized from fermented dairy products and human origin were screened for their Cd biosorption potential using Flame Atomic Absorption Spectroscopy (FAAS)...
October 21, 2016: Environmental Science and Pollution Research International
Duthika M Mallawaaratchy, Susannah Hallal, Ben Russell, Linda Ly, Saeideh Ebrahimkhani, Heng Wei, Richard I Christopherson, Michael E Buckland, Kimberley L Kaufman
Extracellular vesicles (EVs) play key roles in glioblastoma (GBM) biology and represent novel sources of biomarkers that are detectable in the peripheral circulation. Despite this notionally non-invasive approach to assess GBM tumours in situ, a comprehensive GBM EV protein signature has not been described. Here, EVs secreted by six GBM cell lines were isolated and analysed by quantitative high-resolution mass spectrometry. Overall, 844 proteins were identified in the GBM EV proteome, of which 145 proteins were common to EVs secreted by all cell lines examined; included in the curated EV compendium (Vesiclepedia_559; http://microvesicles...
October 21, 2016: Journal of Neuro-oncology
S Jovtchev, S Alexandrov, N Hristova-Avakumova, S Miteva, L Traikov, D Gerasimova, S Stoeff
BACKGROUND: Different colloids are used as a part of solutions for fluid resuscitation and organ preservation: hydroxyethyl starches (HES), dextran (Dx), polyethylene glycols (PEG), polyvinyl pyrrolidone (PVP). Some of the problems associated with their application are addressed to alteration in erythrocyte (ERY) rheology. OBJECTIVE: We intended to estimate in vitro and compare the aggregation power (AP) of these molecules related to ERY interactions. METHODS: Washed human ERY are used during the study...
October 18, 2016: Clinical Hemorheology and Microcirculation
Stephan Brauckmann, Katharina Effenberger-Neidnicht, Herbert de Groot, Michael Nagel, Christian Mayer, Jürgen Peters, Matthias Hartmann
While hemolysis in patients with sepsis is associated with increased mortality its mechanisms are unknown and Toll-like receptor (TLR)-4 mediated effects, complement-mediated hemolysis, or direct cell membrane effects are all conceivable mechanisms. In this study, we tested the hypotheses that toxic lipopolysaccharide (LPS) as well as non-toxic RS-LPS evokes hemolysis (1) by direct membrane effects, and (2) independent of the complement system and TLR-4 activation. We found, that incubation with LPS resulted in a marked time and concentration dependent increase of free hemoglobin concentration and LDH activity in whole blood and washed red cells...
October 19, 2016: Scientific Reports
Imran Rashid, Nathalie Moiré, Bruno Héraut, Isabelle Dimier-Poisson, Marie-Noëlle Mévélec
Infection with the parasite Toxoplasma gondii causes serious public health problems and is of great economic importance worldwide. No vaccine is currently available, so the design of efficient vaccine strategies is still a topical question. In this study, we evaluated the immunoprophylactic potential of a T. gondii virulence factor, the rhoptry kinase ROP18, in a mouse model of chronic toxoplasmosis: first using a recombinant protein produced in Schneider insect cells adjuvanted with poly I:C emulsified in Montanide SV71 by a parenteral route or adjuvanted with cholera toxin by the nasal route and second using a DNA plasmid encoding ROP18 adjuvanted with GM-CSF ± IL-12 DNA...
October 18, 2016: Medical Microbiology and Immunology
Asha Bonney, Michael Christie, Anne Beaty, Sebastian Lunke, Graham Taylor, Louis Irving, Daniel Steinfort
BACKGROUND: With the rapid development of genotype-guided therapies, molecular testing is becoming important in the management of lung cancer. Bronchoscopy is one of the most common investigations performed to diagnose and investigate lung cancer. Given the limited samples often produced by bronchoscopy, this study aims to evaluate the feasibility of performing molecular testing on cell blocks created from bronchoscope cytology brush tip washings (BTW). METHODS: Patients with positive brush cytology for tumour cells had cell blocks created from the BTW...
September 2016: Journal of Thoracic Disease
Amit Modgil, Manasa L Parakala, Mike A Ackley, James J Doherty, Stephen J Moss, Paul A Davies
The neuroactive steroid (NAS) tetrahydrodeoxycorticosterone (THDOC) increases protein kinase C (PKC) mediated phosphorylation of extrasynaptic GABAA receptor (GABAAR) subunits leading to increased surface expression of α4/β3 subunit-containing extrasynaptic GABAARs, leading to a sustained increase in GABAAR tonic current density. Whether other naturally occurring and synthetic NASs share both an allosteric and metabotropic action on GABAARs is unknown. Here, we examine the allosteric and metabotropic properties of allopregnanolone (ALLO), and synthetic NASs SGE-516 and ganaxolone...
October 12, 2016: Neuropharmacology
Amanda N Fader, James Java, Meaghan Tenney, Stephanie Ricci, Camille C Gunderson, Sarah M Temkin, Nick Spirtos, Christina L Kushnir, Michael L Pearl, Oliver Zivanovic, Krishnansu S Tewari, David O'Malley, Ellen M Hartenbach, Chad A Hamilton, Natalie S Gould, Robert S Mannel, William Rodgers, Joan L Walker
OBJECTIVES: We sought to analyze the clinicopathologic features, recurrence patterns and survival outcomes of women with high-grade uterine cancer (UC) enrolled on The Gynecologic Oncology Group (GOG) LAP2 trial. METHODS: This is a post-hoc analysis of LAP-2 patients with grade 3 endometrioid adenocarcinoma (ENDO), uterine serous (USC), clear cell (CC) and carcinosarcoma (CS). Demographics, clinicopathologic features, and recurrence patterns, were compared by histology and surgical approach...
October 12, 2016: Gynecologic Oncology
Janine Stucke-Ring, Julian Ronnacker, Caroline Brand, Carsten Höltke, Christoph Schliemann, Torsten Kessler, Lars Henning Schmidt, Saliha Harrach, Verena Mantke, Heike Hintelmann, Wolfgang Hartmann, Eva Wardelmann, Georg Lenz, Bernhard Wünsch, Carsten Müller-Tidow, Rolf M Mesters, Christian Schwöppe, Wolfgang E Berdel
Truncated tissue factor (tTF), retargeted to tumor vasculature by GNGRAHA peptide (tTF-NGR), and doxorubicin have therapeutic activity against a variety of tumors. We report on combination experiments of both drugs using different schedules. We have tested fluorescence- and HPLC-based intratumoral pharmacokinetics of doxorubicin, flow cytometry for cellular phosphatidylserine (PS) expression, and tumor xenograft studies for showing in vivo apoptosis, proliferation decrease, and tumor shrinkage upon combination therapy with doxorubicin and induced tumor vascular infarction...
October 11, 2016: Oncotarget
N A Jarvis, J R Donaldson, C A O'Bryan, S C Ricke, P G Crandall
Listeria monocytogenes can be carried by and infect poultry, although the clinical disease in birds is rare. Escape from macrophage phagocytosis is a key step in pathogenesis for L. monocytogenes Therefore, we investigated the infection of the chicken macrophage-like cell line HD11 with 2 strains of L. monocytogenes EGD-e and Scott A. After infection, L. monocytogenes was quantified by spread plating and HD11 was quantified with trypan blue exclusion stain before enumeration. The standard macrophage killing protocols require washing the cell monolayers 3 times with PBS, which was found to negatively influence HD11 monolayers...
October 12, 2016: Poultry Science
Roman Štukelj, Karin Schara, Apolonija Bedina-Zavec, Vid Šuštar, Manca Pajnič, Ljubiša Pađen, Judita Lea Krek, Veronika Kralj-Iglič, Anita Mrvar-Brečko, Rado Janša
During harvesting of nanovesicles (NVs) from blood, blood cells and other particles in blood are exposed to mechanical forces which may cause activation of platelets, changes of membrane properties, cell deformation and shedding of membrane fragments. We report on the effect of shear forces imposed upon blood samples during the harvesting process, on the concentration of membrane nanovesicles in isolates from blood. Mathematical models of blood flow through the needle during sampling with vacutubes and with free flow were constructed, starting from the Navier-Stokes formalism...
October 10, 2016: European Journal of Pharmaceutical Sciences
Luiz Alexandre Chisini, Marcus Cristian Muniz Conde, Jose Carlos Bernedo Alcázar, Adriana Fernandes da Silva, Jacques Eduardo Nör, Sandra Beatriz Chaves Tarquinio, Flávio Fernando Demarco
The aim of the present study was to evaluate the expression of transforming growth factor-β1 (TGF-β1) and osteonectin (ON) in pulp-like tissues developed by tissue engineering and to compare it with the expression of these proteins in pulps treated with Ca(OH)2 therapy. Tooth slices were obtained from non-carious human third molars under sterile procedures. The residual periodontal and pulp soft tissues were removed. Empty pulp spaces of the tooth slice were filled with sodium chloride particles (250-425 µm)...
October 10, 2016: Brazilian Oral Research
Siroon Bekkering, Bastiaan A Blok, Leo A B Joosten, Niels P Riksen, Reinout van Crevel, Mihai G Netea
BACKGROUND: Innate immune memory or 'trained immunity' has recently been described as an important property of cells of the innate immune system. Due to the increased interest in this important new field of immunological investigation we sought to determine the optimal conditions for an in-vitro experimental protocol of monocyte training using three of the most commonly used training stimuli from the literature: β-glucan, the Bacille Calmette-Guérin (BCG) vaccine, and oxidized LDL (oxLDL)...
October 12, 2016: Clinical and Vaccine Immunology: CVI
R Viter, K Jekabsons, Z Kalnina, N Poletaev, S H Hsu, U Riekstina
Using photoluminescent ZnO nanorods and carbohydrate marker SSEA-4, a novel cancer cell recognition system was developed. Immobilization of SSEA-4 antibodies (αSSEA-4) on ZnO nanorods was performed in buffer solution (pH = 7.1) over 2 h. The cancer cell line probes were fixed on the glass slide. One hundred microliters of ZnO-αSSEA-4 conjugates were deposited on the cell probe and exposed for 30 min. After washing photoluminescence spectra were recorded. Based on the developed methodology, ZnO-αSSEA-4 probes were tested on patient-derived breast and colorectal carcinoma cells...
October 12, 2016: Nanotechnology
Asako Tajima, Isha Pradhan, Xuehui Geng, Massimo Trucco, Yong Fan
One of the hallmarks of modern medicine is the development of therapeutics that can modulate immune responses, especially the adaptive arm of immunity, for disease intervention and prevention. While tremendous progress has been made in the past decades, manipulating the thymus, the primary lymphoid organ responsible for the development and education of T lymphocytes, remains a challenge. One of the major obstacles is the difficulty to reproduce its unique extracellular matrix (ECM) microenvironment that is essential for maintaining the function and survival of thymic epithelial cells (TECs), the predominant population of cells in the thymic stroma...
October 12, 2016: Methods in Molecular Biology
Faidad Khan, Xueqing Wu, Gideon L Matzkin, Mohsin A Khan, Fuminori Sakai, Jorge E Vidal
Staphylococcus aureus (Sau) strains are a main cause of disease, including nosocomial infections which have been linked to the production of biofilms and the propagation of antibiotic resistance strains such as methicillin-resistant Staphylococcus aureus (MRSA). A previous study found that Streptococcus pneumoniae (Spn) strains kill planktonic cultures of Sau strains. In this work, we have further evaluated in detail the eradication of Sau biofilms and investigated ultrastructural interactions of the biofilmicidal effect...
2016: Frontiers in Cellular and Infection Microbiology
Di Sun, Cong Wei, Yi Li, Qijie Lu, Wei Zhang, Bing Hu
Totally 85 patients with 93 renal lesions who underwent contrast-enhanced ultrasound (CEUS) were retrospectively studied with quantitative analysis to evaluate its value in the differential diagnosis of renal tumor histotypes. CEUS characteristics were analysed including the enhancement patterns, peak intensity, homogeneity of enhancement, and pseudocapsule. Quantitative parameters of peak intensity (P) and time to peak (TP) were measured with QontraXt software, and the index "relative enhancement percentage" ΔP% and "difference in TP between tumor and cortex" ΔTP were used to quantify the CEUS features of renal tumors...
October 11, 2016: Scientific Reports
Claudio D'Amore, Genny Orso, Fabio Fusi, Mario A Pagano, Giovanni Miotto, Alessia Forgiarini, Sara De Martin, Giulia Castellani, Giovanni Ribaudo, David Rennison, Margaret A Brimble, Brian Hopkins, Alessandro Ferrarese, Sergio Bova
Norbormide (NRB) is a unique compound that acts directly on rat vascular myocytes to trigger a contractile process, through an as yet unknown mechanism, which results in the selective contraction of rat peripheral arteries. To gain insight into the mechanisms involved in NRB rat-selective activity, we investigated the subcellular distribution of NRB-AF12, a nitrobenzoxadiazole (NBD)-derivative of NRB, in living NRB-sensitive and NRB-insensitive cells. In both cell types, NRB-AF12 localized to the endoplasmic reticulum (ER), Golgi apparatus, mitochondria, lysosomes, and endosomes; however, in NRB-sensitive cells, the fluorescence also extended to the plasma membrane...
2016: Frontiers in Pharmacology
Stephanie Karmann, Stéphanie Follonier, Monica Bassas-Galia, Sven Panke, Manfred Zinn
Poly(3-hydroxyalkanoates) (PHAs) are bio-based and biodegradable polyesters which have been considered as a promising alternative to petrol-based plastics. Its bacterial production is a dynamic process in which intracellular polymerization and depolymerization are closely linked and depend on the availability of carbon substrates and other nutrients. These dynamics require a fast and quantitative method to determine the optimal harvest-time of PHA containing cells or to adjust carbon supply. In principle, flow cytometry (FCM) is an ideal tool that suits these requirements and, in addition, provides data on the PHA content of different cell populations...
October 6, 2016: Journal of Microbiological Methods
Masayuki Shintaku, Makoto Dohi, Yoshihiro Yamamoto, Aya Nagai, Toshihiro Higuchi
We report a case of clear cell carcinoma (CCC) of the ovary with plasma cell-rich inflammatory stroma, a recently proposed subtype of CCC, and present the cytological findings. The patient was a 48-year-old woman, who was incidentally found to have a right ovarian tumor during the preoperative work-up for an early-stage adenocarcinoma of the uterine cervix. Cytological examination of an imprint smear of the ovarian tumor and peritoneal washing revealed solid cell clusters of irregular, often dendritic shapes, which were intermingled with many inflammatory cells...
October 8, 2016: Diagnostic Cytopathology
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