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Cell washing

Chenyu Wang, Wenwen Liu, Qingquan Wei, Lufeng Ren, Manqing Tan, Yude Yu
Conventional cell-sized well arrays have advantages of high occupancy, simple operation, and low cost for capturing single-cells. However, they have insufficient space for including reagents required for cell treatment or analysis, which restricts the wide application of cell-sized well arrays as a single-cell research tool alone. Here, we present a novel dual-well array chip, which integrates capture-wells (20  μ m in diameter) with reaction-wells (100  μ m in diameter) and describe a flow method for convenient single-cell analysis requiring neither complicated infra-structure nor high expenditure, while enabling highly efficient single cell trapping (75...
May 2018: Biomicrofluidics
Ajay Sheshadri, Dimpy P Shah, Myrna Godoy, Jeremy J Erasmus, Juhee Song, Liang Li, Scott E Evans, Roy F Chemaly, Burton F Dickey, David E Ost
BACKGROUND: Radiologic severity may predict adverse outcomes after lower respiratory tract infection (LRI). However, few studies have quantified radiologic severity of LRIs. We sought to evaluate whether a semi-quantitative scoring tool, the Radiologic Severity Index (RSI), predicted mortality after parainfluenza virus (PIV)-associated LRI. METHODS: We conducted a retrospective review of consecutively-enrolled adult patients with hematologic malignancy or hematopoietic stem cell transplantation and with PIV detected in nasal wash who subsequently developed radiologically-confirmed LRI...
2018: PloS One
Nira M Soren, Anil K Sharma, Vadali R B Sastry
The study investigated the long-term effect of feeding processed solvent extracted karanj ( Pongamia glabra ) cake (SKC) on gross pathology and histopathological changes in some vital organs, and on the activities of serum enzymes in Jalauni lambs. Twenty-four male lambs were divided into 4 groups and allotted randomly to a soybean meal (SBM) based control (CON) and 3 treatment groups receiving concentrate mixtures, containing water washed (WW), 2.5% lime (LM) and 0.4% binder (BN) treated SKC replacing 50% nitrogen of SBM to meet the protein requirements...
June 2017: Animal nutrition
Barbara Marzani, Daniela Pinto, Elisabetta Sorbellini, Fabio Rinaldi
BACKGROUND: Considering the importance of hair in our modern society and the impact of hair loss, the efforts of researchers are addressed to better understand the mechanisms behind the hair cycle regulation and dysregulation. Because hair loss is multifactorial, differenced and new approaches are required. In particular we addressed our attention to two recently identified targets in hair cycling and growth control: olfactory receptor and autophagy. The aim of the study was to evaluate: the possible pro-autophagic effect of N1-methylspermidine (a spermidine analogue) in vitro and, in a double blind clinical trial, the safety and efficacy of topical daily application of a lotion containing N1-methylspermidine and Sandalore®...
June 2018: Giornale Italiano di Dermatologia e Venereologia: Organo Ufficiale, Società Italiana di Dermatologia e Sifilografia
Alizé Proust, Julie-Christine Lévesque, Corinne Barat, Sachiko Sato, Michel J Tremblay
Extracellular traps ejected by various immune cells (neutrophils, macrophages, eosinophils and mast cells) have several immune functions, either protective against pathogens or deleterious in some autoimmune or inflammatory disorders. Since their first description in 2004, the mechanisms of extracellular traps formation have been extensively investigated though still not fully understood. We describe here a new tool for the detection of extracellular traps by fluorescence microscopy in a single-step staining protocol, which does not require any wash...
May 16, 2018: Methods and Applications in Fluorescence
Christiane Reitz
In recent years, genomic, animal and cell biology studies have implicated deficiencies in retromer-mediated trafficking of proteins in an increasing number of neurodegenerative diseases including Alzheimer's Disease (AD), Parkinson's Disease (PD) and Frontotemporal Lobar Degener-ation (FTLD). The retromer complex, which is highly conserved across all eukaryotes, regulates the sorting of transmembrane proteins out of endo-somes to the cell surface or to the trans-Golgi network. Within retromer, cargo selection and binding are performed by a trimer of the Vps26, Vps29 and Vps35 proteins, named the "Cargo-Selective Complex (CSC)"...
May 2018: Current Genomics
Sebastian Hauke, Kaya Keutler, Prasad Phapale, Dmytro A Yushchenko, Carsten Schultz
The secretion of insulin from β -cells depends on extracellular factors, in particular glucose and other small molecules, some of which act on G-protein coupled receptors (GPCRs). Fatty acids (FAs) have been discussed as exogenous secretagogues of insulin for decades, especially after the FA receptor GPR40 was discovered. However, the role of FAs as endogenous signaling factor has not been investigated until now. In the present work, we demonstrate that lowering endogenous FA levels in β -cell medium by stringent washing or by the application of fatty acid-free bovine serum albumin (FAF-BSA) immediately reduced glucose-induced oscillations of cytosolic Ca2+ ([Ca2+ ]i oscillations) in MIN6 and primary β -cells, as well as insulin secretion...
May 10, 2018: Diabetes
Sinéad Knight, Helen Plant, Lisa McWilliams, Mark Wigglesworth
Adaptation of phenotypic cell assays to 1536-well format brings major challenges in liquid handling for high-content assays requiring washing steps and coating of plates. In addition, problematic edge effects and reduced assay quality are frequently encountered. In this chapter, we describe the novel application of a centrifugal plate washer to facilitate miniaturization of 1536-well cell assays and a combination of techniques to reduce edge effects, all of which improved throughput and data quality. Cell assays currently limited in throughput because of cost and complex protocols may be enabled by the techniques presented in this chapter...
2018: Methods in Molecular Biology
K Memon, Y Cheng, F Panhwar, Z R Chen, Z Haider, S Afridi, P Hu, G Zhao
  BACKGROUND: Osmotic shock upon the addition and removal of cryoprotectant agent (CPA) is a major source of cell damage during cryopreservation. OBJECTIVE: Microfluidic device offers a new platform for CPA loading and unloading. The micro scale dimension makes possible to perform a detailed analysis and controllable removal of CPA with many advantages. MATERIALS AND METHODS: A microfluidic device was developed for extracting dimethyl sulfoxide (DMSO) from the sample streamline...
January 2018: Cryo Letters
J Chen, E Biller, M Losos, J Li, D Hamad, L Blower, M Grevenow, J Oakley
BACKGROUND: Red blood cell (RBC) transfusion is associated with the most transfusion-related adverse events (AE). Recent clinical studies showed no significant difference in transfusion-associated mortality between fresh and older RBCs. However, the impact of storage duration as well as irradiation on nonfatal yet much more common complications has not been fully investigated. MATERIALS/METHODS: In this retrospective study of RBC transfusion-associated AEs, a total of 188,562 units of leucocyte-reduced RBCs were transfused in approximately 5·5 years...
May 6, 2018: Vox Sanguinis
R J Laursen, B Alsbjerg, I Vogel, C H Gravholt, H Elbaek, D L Lildballe, P Humaidan, E M Vestergaard
INTRODUCTION: We present a case of an infertile male with 46,XX/46,XYchimerism fathering a child after ICSI procedure. METHODS: Conventional cytogenetic analysis on chromosomes, derived from lymphocytes, using standard Q-banding procedures with a 450-550-band resolution and short-tandem-repeat analysis of 14 loci. RESULTS: Analysis of 20 metaphases from lymphocytes indicated that the proband was a karyotypic mosaic with an almost equal distribution between male and female cell lines...
April 30, 2018: Journal of Assisted Reproduction and Genetics
Cristina Simões Cortinhas, Bruno Garcia Botaro, Susana Nori de Macedo, Marcos Veiga Dos Santos
This study identified the association of management practices and herd characteristics with milk quality of bulk tanks in southeastern, Brazil. Milk samples were collected weekly during 8 weeks from 63 dairy herds. Bulk tanks were evaluated for total bacteria (TBC), preliminary incubation (PIC), pasteurization (PC), coliform (CC), and somatic cell counts (SCC). Associations found were type of milking system utilized in the farm with TBC, PIC, and SCC; the use of gloves for milking with TBC and PIC; sanitation of milking equipment prior to milking with PC and CC; strip cup testing of cows with PC; teat washing prior to milking with SCC; pre-milking teat disinfection with TBC and CC; post-dipping with TBC and SCC; and the alkaline-acid washing procedure of milking equipment with PIC and PC...
April 30, 2018: Tropical Animal Health and Production
Jenna A Walz, Charles R Mace
Immunophenotyping is typically achieved using flow cytometry, but any influence this biomarker may have on adhesion or surface recognition cannot be determined concurrently. In this manuscript, we demonstrate the utility of lateral microscopy for correlating cell surface biomarker expression levels with quantitative descriptions of cell morphology. With our imaging system, we observed single cells from two T cell lines and two B cell lines adhere to antibody-coated substrates and quantified this adhesion using contact angle measurements...
April 30, 2018: Analytical Chemistry
Arvind Kumar Bishnoi, Pankaj Garg, Kartik Patel, Chandrasekaran Ananthanarayanan, Ritesh Shah, Atul Solanki, Himani Pandya, Sanjay Patel
BACKGROUND: Retrospective reviews have found the use of stored packed red blood cells (PRBCs) in priming to be associated with increased risk of postoperative complications. The purpose of study was to prospectively investigate the influence of duration of storage of PRBCs used in priming the cardiopulmonary bypass (CPB) circuit on the metabolic profile of the patients, and postoperative outcome after paediatric cardiac surgery. METHODS: Between January 2015 and December 2015, 198 consecutive children operated for cardiac surgery using CPB and received blood for priming the circuit were included...
March 31, 2018: Heart, Lung & Circulation
(no author information available yet)
No abstract text is available yet for this article.
April 2018: Medicine (Baltimore)
Sara Alicia González Porto, Nieves Domenech, Alba González Rodríguez, Edgar Mauricio Avellaneda Oviedo, Francisco J Blanco, María C Arufe Gonda, Ángel Álvarez Jorge, Jacinto Sánchez Ibañez, Esther Rendal Vázquez
The purpose of the current study was to establish a valid protocol for nerve cryopreservation, and to evaluate if the addition of albumin supposed any advantage in the procedure. We compared a traditional cryopreservation method that uses dimethyl sulfoxide (DMSO) as cryoprotectant, to an alternative method that uses DMSO and albumin. Six Wistar Lewis rats were used to obtain twelve 20 mm fragments of sciatic nerve. In the first group, six fragments were cryopreserved in 199 media with 10% DMSO, with a temperature decreasing rate of 1 °C per minute...
April 26, 2018: Cell and Tissue Banking
Tao Hua, Xuehua Zhang, Bo Tang, Chen Chang, Guoyang Liu, Lei Feng, Yang Yu, Daohua Zhang, Jibo Hou
BACKGROUND: Low concentrations of nonionic surfactants can change the physical properties of cell membranes, and thus and in turn increase drug permeability. Porcine circovirus 2 (PCV2) is an extremely slow-growing virus, and PCV2 infection of PK-15 cells yields very low viral titers. The present study investigates the effect of various nonionic surfactants, namely, Tween-20, Tween-28, Tween-40, Tween-80, Brij-30, Brij-35, NP-40, and Triton X-100 on PCV2 infection and yield in PK-15 cells...
April 24, 2018: BMC Veterinary Research
Pin-Chao Liao, Istvan R Boldogh, Stephanie E Siegmund, Zachary Freyberg, Liza A Pon
Isolated mitochondria are widely used to study the function of the organelle. Typically, mitochondria are prepared using differential centrifugation alone or in conjunction with density gradient ultracentrifugation. However, mitochondria isolated using differential centrifugation contain membrane or organelle contaminants, and further purification of crude mitochondria by density gradient ultracentrifugation requires large amounts of starting material, and is time-consuming. Mitochondria have also been isolated by irreversible binding to antibody-coated magnetic beads...
2018: PloS One
Atsuko Ohki, Nobutsugu Abe, Eri Yoshimoto, Yoshikazu Hashimoto, Hirohisa Takeuchi, Gen Nagao, Tadahiko Masaki, Toshiyuki Mori, Yasuo Ohkura, Masanori Sugiyama
PURPOSE: Intragastric free cancer cells in patients with gastric cancer have rarely been studied. The purpose of this study was to investigate the detection rate of intragastric free cancer cells in gastric washes using two types of solutions during endoscopic examination. We further clarified risk factors affecting the presence of exfoliated free cancer cells. METHODS: A total of 175 patients with gastric cancer were enrolled. Lactated Ringer's solution (N = 89) or distilled water (DW; N = 86) via endoscopic working channel was sprayed onto the tumor surface, and the resultant fluid was collected for cytological examination...
April 25, 2018: Gastric Cancer
Dinesh Shetty, Tina Skorjanc, Jesus Raya, Sudhir Kumar Sharma, Ilma Jahovic, Kyriaki Polychronopoulou, Zouhair Asfari, Dong Suk Han, Sajeewa Dewage, John-Carl Olsen, Ramesh Jagannathan, Serdal Kirmizialtin, Ali Trabolsi
Calixarenes are a common motif in supramolecular chemistry but have rarely been incorporated in structurally well-defined covalent 2D materials. Such a task is challenging, especially without a template, because of the non-planar nature and conformational flexibility of the calixarene ring. Here, we report the first-of-kind solvothermal synthesis of a calix[4]arene-based 2D polymer (CX4-NS), that is porous, covalent, and isolated as few-layer thick (3.52 nm) nanosheets. Experimental and theoretical characterization of the nanosheets are presented...
April 24, 2018: ACS Applied Materials & Interfaces
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