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Makoto Hirano, Kiichiro Totani, Tomohiko Fukuda, Jianguo Gu, Akemi Suzuki
BACKGROUND: Megalin is a 600-kDa single-spanning transmembrane glycoprotein and functions as an endocytic receptor, distributed not only in the kidney but also in other tissues. Structurally and functionally distinct ligands for megalin have been identified. Megalin has 30 potential N-glycosylation sites in its extracellular domain. We found that megalin interacts with its ligands in a glycoform-dependent manner. METHODS: Distribution of megalin and glycans was histochemically analyzed in mouse kidneys...
October 20, 2016: Biochimica et Biophysica Acta
Alan D Stern, Adeeb H Rahman, Marc R Birtwistle
Mass cytometry offers the advantage of allowing the simultaneous measurement of a greater number parameters than conventional flow cytometry. However, to date, mass cytometry has lacked a reliable alternative to the light scatter properties that are commonly used as a cell size metric in flow cytometry (forward scatter intensity-FSC). Here, we report the development of two plasma membrane staining assays to evaluate mammalian cell size in mass cytometry experiments. One is based on wheat germ agglutinin (WGA) staining and the other on Osmium tetroxide (OsO4 ) staining, both of which have preferential affinity for cell membranes...
October 21, 2016: Cytometry. Part A: the Journal of the International Society for Analytical Cytology
Steen Kølvraa, Ripudaman Singh, Elizabeth A Normand, Sadeem Qdaisat, Ignatia B Van denVeyver, Laird Jackson, Lotte Hatt, Palle Schelde, Niels Uldbjerg, Else Marie Vestergaard, Li Zhao, Rui Chen, Chad A Shaw, Amy M Breman, Arthur L Beaudet
OBJECTIVE: Non-invasive prenatal testing (NIPT) based on fetal cells in maternal blood has the advantage over NIPT based on circulating cell-free fetal DNA in that there is no contamination with maternal DNA. This will most likely result in better detection of chromosomal aberrations including subchromosomal defects. The objective of this study was to test whether fetal cells enriched from maternal blood can be used for cell-based NIPT. METHODS: We present a method for enriching fetal cells from maternal blood, subsequent amplification of the fetal genome and detection of chromosomal and subchromosomal variations in the genome...
October 19, 2016: Prenatal Diagnosis
Behjatolah Monzavi-Karbassi, Fariba Jousheghany, Thomas Kieber-Emmons
Development of cancer vaccines targeting tumor-associated antigens (TAAs) is an alternative approach to chemotherapy with sustained anti-tumor effects. The success of active immunotherapy has been hampered by tumor-induced immune suppressors. Regulatory T cells (Tregs) are a population of immune suppressors with a proven role in regulating anti-tumor immune responses. Removing or subduing Tregs activity leads to more robust anti-tumor immune responses. Here, we used a cell-based vaccination strategy in the 4T1 murine mammary model to examine whether bulk removal of certain TAAs, using their glycan profile, can affect the immunogenicity of the vaccine...
October 19, 2016: Immunological Investigations
Evan J Anderson, Xavier Carbonell-Estrany, Maarten Blanken, Marcello Lanari, Margaret Sheridan-Pereira, Barry Rodgers-Gray, John Fullarton, Elisabeth Rouffiac, Pamela Vo, Gerard Notario, Fiona Campbell, Bosco Paes
BACKGROUND: Moderate-late preterm infants, 33-35 weeks' gestational age (wGA), are at increased risk for respiratory syncytial virus hospitalization (RSVH). OBJECTIVE: To quantify the burden of RSVH in moderate-late preterm infants. METHODS: A pooled analysis was conducted on RSVH from 7 prospective, observational studies in the Northern Hemisphere from 2000-2014. Infants' 33 -35 wGA without comorbidity born during the RSV season who did not receive RSV immunoprophylaxis were enrolled...
October 17, 2016: Pediatric Infectious Disease Journal
Yusuke Tonai, Ryu Ishihara, Yasushi Yamasaki, Takashi Kanesaka, Sachiko Yamamoto, Tomofumi Akasaka, Noboru Hanaoka, Yoji Takeuchi, Koji Higashino, Noriya Uedo, Yasuhiko Tomita, Hiroyasu Iishi
Background and study aims: Better endoscopic diagnosis in case of Barrett's esophagus is still needed. White globe appearance (WGA) is a novel endoscopic marker for gastric adenocarcinoma, with high sensitivity for differentiating between gastric cancer/high-grade dysplasia and other lesions. We report 2 cases of esophageal adenocarcinoma with WGA. In Case 1, esophagogastroduodenoscopy (EGD) revealed a 10-mm esophageal adenocarcinoma in a 48-year-old Japanese woman with short-segment Barrett's esophagus. A small (< 1 mm) white globular lesion, typical of WGA, was observed under the epithelium by magnifying narrow-band imaging...
October 2016: Endoscopy International Open
Maria Hammond, Felix Homa, Helene Andersson-Svahn, Thijs J G Ettema, Haakan N Joensson
BACKGROUND: Whole genome amplification (WGA) is a challenging, key step in metagenomic studies of samples containing minute amounts of DNA, such as samples from low biomass environments. It is well known that multiple displacement amplification (MDA), the most commonly used WGA method for microbial samples, skews the genomic representation in the sample. We have combined MDA with droplet microfluidics to perform the reaction in a homogeneous emulsion. Each droplet in this emulsion can be considered an individual reaction chamber, allowing partitioning of the MDA reaction into millions of parallel reactions with only one or very few template molecules per droplet...
October 6, 2016: Microbiome
Jeremy A Franklin, Evan J Anderson, Xionghua Wu, Christopher S Ambrose, Eric A F Simões
Background.  Database studies have identified that public health insurance status is associated with an increased risk of severe respiratory syncytial virus (RSV) disease in US infants. However, these studies did not adjust for the presence of other risk factors and did not evaluate the risk in preterm infants. Methods.  In this study, we evaluate the independent association between public insurance and severe RSV disease outcomes adjusting for other risk factors. The prospective, observational RSV Respiratory Events among Preterm Infants Outcomes and Risk Tracking (REPORT) study was conducted over 2 consecutive RSV seasons at 188 US clinical sites that enrolled preterm infants born at 32-35 wGA who had not received RSV immunoprophylaxis with palivizumab...
September 2016: Open Forum Infectious Diseases
Deep Pooja, Hitesh Kulhari, Madhusudana Kuncha, Shyam Sunder Rachamalla, David J Adams, Vipul Bansal, Ramakrishna Sistla
Oral delivery of anticancer drugs remains challenging despite the most convenient route of drug administration. Hydrophobicity and non-specific toxicities of anticancer agents are major impediments in the development of oral formulation. In this study, we developed wheat germ agglutinin (WGA)-conjugated, solid lipid nanoparticles to improve the oral delivery of the hydrophobic anticancer drug, paclitaxel (PTX). This study was focused to improve the PTX loading in biocompatible lipid matrix with high bioconjugation efficiency...
October 3, 2016: Molecular Pharmaceutics
Juanjuan Xu, Rui Fang, Li Chen, Daozhen Chen, Jian-Ping Xiao, Weimin Yang, Honghua Wang, Xiaoqing Song, Ting Ma, Shiping Bo, Chong Shi, Jun Ren, Lei Huang, Li-Yi Cai, Bing Yao, X Sunney Xie, Sijia Lu
Preimplantation genetic screening (PGS) is widely used to select in vitro-fertilized embryos free of chromosomal abnormalities and to improve the clinical outcome of in vitro fertilization (IVF). A disadvantage of PGS is that it requires biopsy of the preimplantation human embryo, which can limit the clinical applicability of PGS due to the invasiveness and complexity of the process. Here, we present and validate a noninvasive chromosome screening (NICS) method based on sequencing the genomic DNA secreted into the culture medium from the human blastocyst...
September 29, 2016: Proceedings of the National Academy of Sciences of the United States of America
Juan Wang, Lian-Guo Zhang, Ning Li, Ming Lei, Yun Ma, Yuan-Hong Peng, Wei-Hua Bian
OBJECTIVE: To construct a lentivirus vector carrying wheat germ agglutinin (WGA) and evaluate its ability of tracing WGA in the brain of mice with ischemic brain injury. METHODS: WGA gene was inserted into the lentiviral vector Plvx IRES-ZsGreen1 using genetic engineering methods. 293T cells were transfected with the vector and 3 packaging plasmids (RPEV, PRRE, and VSVG) to obtain the recombinant lentivirus for infection of human adipose-derived stem cells (hADSCs)...
August 20, 2016: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
Trina Manalo, Adam May, Joshua Quinn, Dominique S Lafontant, Olubusola Shifatu, Wei He, Juan M Gonzalez-Rosa, Geoffrey C Burns, Caroline E Burns, Alan R Burns, Pascal J Lafontant
Lectins are carbohydrate-binding proteins commonly used as biochemical and histochemical tools to study glycoconjugate (glycoproteins, glycolipids) expression patterns in cells, tissues, including mammalian hearts. However, lectins have received little attention in zebrafish (Danio rerio) and giant danio (Devario aequipinnatus) heart studies. Here, we sought to determine the binding patterns of six commonly used lectins-wheat germ agglutinin (WGA), Ulex europaeus agglutinin, Bandeiraea simplicifolia lectin (BS lectin), concanavalin A (Con A), Ricinus communis agglutinin I (RCA I), and Lycopersicon esculentum agglutinin (tomato lectin)-in these hearts...
September 28, 2016: Journal of Histochemistry and Cytochemistry: Official Journal of the Histochemistry Society
Hans Yu, Judith Reiser, Urban Besenfelder, Ebrahim Razzazi-Fazeli, Jonas Bergquist, Gottfried Brem, Konstantin Artemenko, Corina Mayrhofer
The analysis of glycoproteins in body fluids represents a central task in the study of vital processes. Herein we assessed the combined use of Concanavalin A (ConA) and Wheat Germ Agglutinin (WGA) as ligands to fractionate and enrich glycoproteins from oviductal fluid (OF) that is a source of molecules involved in fertilization. First, the selectivity was corroborated by a gel-based approach using glycoprotein staining and enzymatic deglycosylation. Nano-liquid chromatography-tandem mass spectrometry (nLC-ESI-MS/MS) further on allowed the reliable identification of 134 non-bound as well as 130 lectin bound OF proteins...
September 28, 2016: Proteomics
Tadas Povilaitis, Gediminas Alzbutas, Rasa Sukackaite, Juozas Siurkus, Remigijus Skirgaila
Compartmentalized self replication (CSR) is widely used for in vitro evolution of thermostable DNA polymerases able to perform PCR in emulsion. We have modified and adapted CSR technique for isothermal DNA amplification using mezophilic phi29 DNA polymerase and whole genome amplification (WGA) reaction. In standard CSR emulsified bacterial cells are disrupted during denaturation step (94-96°C) in the first circles of PCR. Released plasmid DNA that encodes target polymerase and the thermophilic enzyme complement the emulsified PCR reaction mixture and start polymerase gene amplification...
September 26, 2016: Protein Engineering, Design & Selection: PEDS
Zaki Hakami, Satoshi Wakisaka
The development of submucosal glands of rat nasopharynx was studied with respect to their morphological maturation and glycoprotein alterations during the postnatal period. This study examined the histological morphology with hematoxylin-eosin and the binding pattern of lectins, soybean agglutinin (SBA), Dolichos biflorus agglutinin (DBA), Vicia villosa agglutinin (VVA), Ulex europaeus agglutinin-I (UEA-I), peanut agglutinin (PNA), wheat germ agglutinin (WGA), and succinylated WGA (sucWGA) on frozen sections from newborn into adulthood...
September 23, 2016: Acta Histochemica
Albert Lee, Devin Miller, Roger Henry, Venkata D P Paruchuri, Robert N O'Meally, Tatiana Boronina, Robert N Cole, Natasha E Zachara
O-Linked N-acetyl-β-d-glucosamine (O-GlcNAc) is a dynamic post-translational modification that modifies and regulates over 3000 nuclear, cytoplasmic, and mitochondrial proteins. Upon exposure to stress and injury, cells and tissues increase the O-GlcNAc modification, or O-GlcNAcylation, of numerous proteins promoting the cellular stress response and thus survival. The aim of this study was to identify proteins that are differentially O-GlcNAcylated upon acute oxidative stress (H2O2) to provide insight into the mechanisms by which O-GlcNAc promotes survival...
October 14, 2016: Journal of Proteome Research
Paolo Manzoni, Bosco Paes, Krista L Lanctôt, Alberto DallʼAgnola, Ian Mitchell, Sara Calabrese, Milena Maule, Elisa Girardi, Tetsuhiro Harimoto, Abby Li
BACKGROUND: Respiratory syncytial virus (RSV) infection frequently results in RSV-related hospitalization (RSVH) in young infants. We examined the outcomes of palivizumab recipients within the Canadian Registry (CARESS) and the Torino-Verona Italian Registry over the 2002-2014 RSV seasons. METHODS: RSVHs were captured during the study seasons. Premature infants who received palivizumab (≤35 completed weeks gestational age (wGA); Group1) were compared with infants given palivizumab for underlying disorders regardless of gestational age (Group2)...
September 19, 2016: Pediatric Infectious Disease Journal
Josep Figueras-Aloy, Paolo Manzoni, Bosco Paes, Eric A F Simões, Louis Bont, Paul A Checchia, Brigitte Fauroux, Xavier Carbonell-Estrany
INTRODUCTION: The REGAL (RSV Evidence-a Geographical Archive of the Literature) series provide a comprehensive review of the published evidence in the field of respiratory syncytial virus (RSV) in Western countries over the last 20 years. This second publication covers the risk and burden of RSV infection in preterm infants born at <37 weeks' gestational age (wGA) without chronic lung disease or congenital heart disease. METHODS: A systematic review was undertaken for articles published between January 1, 1995 and December 31, 2015...
September 14, 2016: Infectious Diseases and Therapy
Dilek Akşit, Alper Yazıcı, Hasan Akşit, Esin S Sarı, Arzu Yay, Onur Yıldız, Adil Kılıç, Sıtkı S Ermiş, Kamil Seyrek
BACKGROUND: Investigate alterations in the expression and localization of carbohydrate units in rat retinal cells exposed to cisplatin toxicity. AIMS: The aim of the study was to evaluate putative protective effects of selenium on retinal cells subjected to cisplatin. STUDY DESIGN: Animal experiment. METHODS: Eighteen healthy Wistar rats were divided into three equal groups: 1. Control, 2. Cisplatin and 3. Cisplatin+selenium groups...
July 2016: Balkan Medical Journal
G P Pinto, K A R Lopes, N G Salles, C Pacheco-Soares
The use of lectins can play an important role for tracking modification on cell surface components, since lectins can be easily complexed with radioisotopes, biotin or fluorescein, facilitating the evaluation of carbohydrates distribution in the cell and mitochondrial activity. The aim of this study was to evaluate photodynamic therapy effects on indirect distribution of N-acetyl-glucosamine terminal glycoproteins, in human laryngeal carcinoma HEp-2 cell line surface, using lectin wheat germ agglutinin (WGA) and on mitochondrial activity, for the same cell line, using MitoTracker...
September 5, 2016: Journal of Fluorescence
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