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Protein electrophoresis

Deniz Turan, Monika Gibis, Gurbuz Gunes, Stefan K Baier, Jochen Weiss
The conjugation reaction of electrospun fibers of a mixture of whey protein isolate (WPI) and dextran using different molecular weights (40, 70, and 100 kDa) and mixing ratios was studied. This study includes the electrospinnability of a mixture of WPI and dextran, and the conjugation reaction between them via the initial stage of the Maillard reaction. The WPI-dextran fibers were characterized using optical and transmission electron microscopy. The covalent attachment of dextran to WPI was confirmed using sodium-dodecyl-sulfate-polyacrylamide gel-electrophoresis with protein and glycoprotein staining...
March 15, 2018: Food & Function
Héctor Miranda-Astudillo, Lilia Colina-Tenorio, Alejandra Jiménez-Suárez, Miriam Vázquez-Acevedo, Bénédicte Salin, Marie-France Giraud, Claire Remacle, Pierre Cardol, Diego González-Halphen
The proposal that the respiratory complexes can associate with each other in larger structures named supercomplexes (SC) is generally accepted. In the last decade's most of the data about this association came from studies in yeasts, mammals and plants, and information is scarce in other lineages. Here we studied the supramolecular association of the F1 FO -ATP synthase (complex V) and the respiratory complexes I, III and IV of the colorless alga Polytomella sp. with an approach that involves solubilization using mild detergents, n-dodecyl-β-D-maltoside (DDM) or digitonin, followed by separation of native protein complexes by electrophoresis (BN-PAGE), after which we identified oligomeric forms of complex V (mainly V2 and V4 ) and different respiratory supercomplexes (I/IV6 , I/III4 , I/IV)...
March 11, 2018: Biochimica et Biophysica Acta
Qiang Wang, Akram A Da'dara, Patrick J Skelly
BACKGROUND: Schistosomes are blood dwelling parasitic worms that cause the debilitating disease schistosomiasis. Here we examined the influence of the parasites on their external environment by monitoring the impact of adult Schistosoma mansoni worms on the murine plasma proteome in vitro and, in particular, on how the worms affect the blood coagulation protein high molecular weight kininogen (HK). METHODS: Following the incubation of adult schistosomes in murine plasma, two-dimensional differential in-gel electrophoresis (2D-DIGE) was conducted to look for changes in the plasma proteome compared with control plasma...
March 14, 2018: Parasites & Vectors
Christian Schwedler, Thomas Häupl, Ulrich Kalus, Véronique Blanchard, Gerd-Rüdiger Burmester, Denis Poddubnyy, Berthold Hoppe
BACKGROUND: Galactosylation of immunoglobulin G (IgG) is reduced in rheumatoid arthritis (RA) and assumed to correlate with inflammation and altered humoral immunity. IgG hypogalactosylation also increases with age. To investigate dependencies in more detail, we compared IgG hypogalactosylation between patients with RA, patients with axial spondyloarthritis (axSpA), and healthy control subjects (HC), and we studied it in RA on the background of HLA-DRB1 shared epitope (SE), anticitrullinated protein antibodies (ACPA), and/or rheumatoid factor (RF) status...
March 14, 2018: Arthritis Research & Therapy
Stephanie J Attarian, Sandra L Leibel, Ping Yang, Danielle N Alfano, Brian P Hackett, F Sessions Cole, Aaron Hamvas
BACKGROUND: Mutations in the NKX2-1 gene are associated with lung disease in infants and children. We hypothesize that disruption of normal surfactant gene expression with these mutations contributes to the respiratory phenotypes observed. METHODS: To assess transactivational activity, co-transfection of luciferase reporter vectors containing surfactant protein B or C (SFTPB or SFTPC) promoters with NKX2-1 plasmids was performed and luciferase activity measured...
March 14, 2018: Pediatric Research
Sellappan Selvaraju, Sivashanmugam Parthipan, Lakshminarayana Somashekar, B Krishnan Binsila, Atul P Kolte, Arunachalam Arangasamy, Janivara Parameshwaraiah Ravindra, Stephen A Krawetz
With artificial insemination (AI) and other precision dependent assisted reproductive technologies (ART) being followed in large scale in human and animal reproduction, assessing semen quality and fertilizability is under continuous scrutiny. Various tests have been developed to predict semen quality, but so far no single, highly reliable test is available. In this regard, transcriptomic profiling of spermatozoa assumes significance as it carries the information about spermatogenesis, sperm function, and paternal roles in post-fertilization events...
March 14, 2018: Systems Biology in Reproductive Medicine
Manjunath Siddaramaiah, BolaSadashiva Satish Rao, Manjunath B Joshi, Anirbit Datta, S Sandya, Vasudha Vishnumurthy, Subhash Chandra, Subramanya G Nayak, Kapaettu Satyamoorthy, Krishna Kishore Mahato
In the present study, we have designed a laser induced fluorescence (LIF) based instrumentation and developed a sensitive methodology for the effective separation, visualization, identification and analysis of proteins on a single platform. In this method, intrinsic fluorescence spectra of proteins were detected after separation on one or two dimensional Sodium Dodecyl Sulfate-Tris(2-carboxyethyl)phosphine (SDS-TCEP) polyacrylamide gel electrophoresis (PAGE) and the data were analyzed. The MATLAB assisted software was designed for the development of PAGE fingerprint for the visualization of protein after one and two-dimensional protein separation...
March 14, 2018: Journal of Biophotonics
Sineepat Talungchit, Waranun Buajeeb, Chotima Lerdtripop, Rudee Surarit, Kongthawat Chairatvit, Sittiruk Roytrakul, Hiroaki Kobayashi, Yuichi Izumi, Siribang-On Piboonniyom Khovidhunkit
BACKGROUND: Salivary protein biomarkers for screening and diagnosis of oral lichen planus (OLP) are not well-defined. The objective of this study was to identify putative protein biomarkers for OLP using proteomic approaches. METHODS: Pooled unstimulated whole saliva was collected from five OLP patients and five healthy control participants. Saliva samples were then subjected to two-dimensional gel electrophoresis, followed by mass spectrometry to identify putative protein biomarkers...
March 13, 2018: BMC Oral Health
M A Afanasyeva, A S Ustiugova, S A Golyshev, A T Kopylov, A V Bogolyubova, D E Demin, P V Belousov, A M Schwartz
Ultracentrifugation on a density gradient remains the only reliable way to obtain highly pure mitochondria preparations. However, it is not readily available for any laboratory and has a serious disadvantage of providing low mitochondria yield, which can be critical when working with limited starting material. Here we describe a combined method for isolation of mitochondria for proteomic studies that includes cell disruption by sonication, differential centrifugation, and magnetic separation. Our method provides remarkable enrichment of mitochondrial proteins as compared to differential centrifugation, magnetic separation, or their combination, and it enables the strongest depletion of cytoplasmic components, as assessed by two-dimensional electrophoresis, mass spectrometry, and Western blot...
January 2018: Biochemistry. Biokhimii︠a︡
A Mostek, M Slowinska, S Judycka, H Karol, A Ciereszko, M A Dietrich
During semen cryopreservation, spermatozoa are exposed to physical and chemical stressors that result in their functional and structural damage. Growing evidence suggests that most cryoinjuries result from oxidative stress accompanying sperm cryopreservation. Elevated amounts of reactive oxygen species (ROS) generated during cryopreservation can react with sperm macromolecules, including proteins. The goal of this study was to investigate the oxidative modifications (measured as carbonylation level changes) of carp spermatozoa proteins triggered by the cryopreservation process...
March 9, 2018: Journal of Animal Science
Ye Liang, Wenhua Xu, Shihai Liu, Jingwei Chi, Jisheng Zhang, Aihua Sui, Liping Wang, Zhijuan Liang, Dan Li, Yuanbin Chen, Haitao Niu
BACKGROUND/AIMS: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a potential anti-cancer agent due to its selective toxicity. However, many human non-small cell lung cancer (NSCLC) cells are partially resistant to TRAIL, thereby limiting its clinical application. Therefore, there is a need for the development of novel adjuvant therapeutic agents to be used in combination with TRAIL. METHODS: In this study, the effect of N-acetyl-glucosamine (GlcNAc), a type of monosaccharide derived from chitosan, combined with TRAIL was evaluated in vitro and in vivo...
March 7, 2018: Cellular Physiology and Biochemistry
Weiqing Qian, Yan Su, Yajie Zhang, Nianwei Yao, Nin Gu, Xu Zhang, Hong Yin
Osteoporosis is a serious public health problem and icariin (ICA) is the active component of the Epimedium sagittatum, a traditional Chinese medicinal herb. The present study aimed to investigate the effects and underlying mechanisms of ICA as a potential therapy for osteoporosis. Calvaria osteoblasts were isolated from newborn rats and treated with ICA. Cell viability, apoptosis, alkaline phosphatase activity and calcium deposition were analyzed. Bioinformatics analyses were performed to identify differentially expressed proteins (DEPs) in response to ICA treatment...
March 9, 2018: Molecular Medicine Reports
Jiaxing Zhou, Dongmiao Liu, Xiong Deng, Shoumin Zhen, Zhimin Wang, Yueming Yan
BACKGROUND: Water deficiency affects grain proteome dynamics and storage protein compositions, resulting in alterations in gluten viscoelasticity. In this study, we investigated the effects of field water deficit on wheat breadmaking quality and grain storage proteins. RESULTS: Water deficiency produced a shorten grain-filling period and a decrease in grain number, grain weight and grain yield, a reduced starch granule; increased protein content and glutenin macropolymer contents, resulting in superior dough properties and breadmaking quality...
March 12, 2018: Journal of the Science of Food and Agriculture
Christina Boeck, Sophia Pfister, Alexander Bürkle, Valerie Vanhooren, Claude Libert, Juan Salinas-Manrique, Detlef E Dietrich, Iris-Tatjana Kolassa, Alexander Karabatsiakis
BACKGROUND: Glycans are short chains of saccharides linked to glycoproteins that are known to be involved in a wide range of inflammatory processes. As depression has been consistently associated with chronic low-grade inflammation, we asked whether patients with Major Depressive Disorder show alterations in the N-glycosylation pattern of serum proteins that might be linked to associated changes in inflammatory processes. METHODS: In a study cohort of 21 female patients with an acute depressive episode and 21 non-depressed female control subjects aged between 50 and 69 years, we analyzed the serum N-glycan profile by DNA Sequencer Adapted-Fluorophore Assisted Carbohydrate Electrophoresis (DSA-FACE) and assessed the serum levels of interleukin (IL)- 6, tumor necrosis factor (TNF)-α and C-reactive protein (CRP) by chemiluminescence immunoassays and nephelometry...
February 27, 2018: Journal of Affective Disorders
Mehdi Moini
In the past few years, there has been a significant effort by the forensic science community to develop new scientific techniques for the analysis of forensic evidence. Forensic chemists have been spearheaded to develop information-rich confirmatory technologies and techniques and apply them to a broad array of forensic challenges. The purpose of these confirmatory techniques is to provide alternatives to presumptive techniques that rely on data such as color changes, pattern matching, or retention time alone, which are prone to more false positives...
March 12, 2018: Electrophoresis
Sule Keskin, Erkan Yalçin, Hazim Özkaya
The objective of this study was to investigate the effects of storage and granary weevil, Sitophilus granarius (L.; Coleoptera: Curculionidae), infestation on pH, protein solubility (PS) and gel electrophoresis properties of meal and roller-milled flours of hard (Ceyhan-99 cv.) and soft (Eser cv.) wheat cultivars, respectively, after 6 mo of storage. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) technique was applied for studying the electrophoretic properties. Hard and soft wheats were infested with non-sexed S...
February 24, 2018: Journal of Economic Entomology
Karolina Nowicka-Bauer, Małgorzata Ozgo, Adam Lepczynski, Marzena Kamieniczna, Agnieszka Malcher, Wiesław Skrzypczak, Maciej Kurpisz
The aim of this study was to identify and analyse human sperm proteins from normozoospermic men using 2-dimensional electrophoresis (2-DE) and mass spectrometry (MS). We identified 73 different sperm proteins, including two less characterized human sperm proteins, Annexin A7 (ANXA7) and c14orf105. Bioinformatic analysis of detected sperm proteins revealed new carbohydrate and lipid metabolic pathways, which supply energy to motile sperm. A comparison of our data with available mRNA microarray data from the human testis allows for validation of identified sperm proteins and aids in the recognition of their physiological pathways...
March 8, 2018: Reproductive Biology
Luciana Teixeira de Siqueira, Marcela Silvestre Outtes Wanderley, Roberto Afonso da Silva, Adriana da Silva Andrade Pereira, José Luiz de Lima Filho, Álvaro Antônio Bandeira Ferraz
BACKGROUND: Obesity has been associated with the development of various types of cancer. Biomarker studies may provide molecular level knowledge of the factors involved in this association, improving clinical practice through new methods of prevention and treatment. PURPOSE: The present study aimed to analyze proteins found in the plasma of obese patients prior to and 6 months after bariatric surgery, using body mass index (BMI) and percentage total weight loss (%TWL) to evaluate, in a prospective manner, the effects of weight loss on the regulation of proteins related to the appearance of tumors...
March 10, 2018: Obesity Surgery
Yueping Zhu, Zhenqing Li, Ping Wang, Lisong Shen, Dawei Zhang, Yoshinori Yamaguchi
Capillary electrophoresis (CE) is an effective tool for protein separation and analysis. Compared with capillary gel electrophoresis (CGE), non-gel sieving capillary electrophoresis (NGSCE) processes the superiority on operation, repeatability and automaticity. Herein, we investigated the effect of polymer molecular weight and concentration, electric field strength, and the effective length of the capillary on the separation performance of proteins, and find that (1) polymer with high molecular weight and concentration favors the separation of proteins, although concentrated polymer hinders its injection into the channel of the capillary due to its high viscosity...
March 3, 2018: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Naoya Uchida, Juan J Haro-Mora, Selami Demirci, Atsushi Fujita, Lydia Raines, Matthew M Hsieh, John F Tisdale
A reliable cell line capable of robust in vitro erythroid differentiation would be useful to investigate red blood cell (RBC) biology and genetic strategies for RBC diseases. K562 cells are widely utilized for erythroid differentiation; however, current differentiation methods are insufficient to analyze globin proteins. In this study, we sought to improve erythroid differentiation from K562 cells to enable protein-level globin analysis. K562 cells were exposed to a variety of reagents including hemin, rapamycin, imatinib, and/or decitabine (known erythroid inducers), and cultured in basic culture media or erythropoietin-based differentiation media...
March 7, 2018: Experimental Hematology
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