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Gel electrophoresis

Jennyka Hallewell, Tim Reuter, Kim Stanford, Ed Topp, Trevor W Alexander
The goal of this study was to monitor Shiga toxin-producing Escherichia coli (STEC) serogroups and virulence genes in cattle (n = 30) originating from a closed herd. Fecal samples were collected (1) at weaning, (2) upon arrival to a feedlot, (3) after 30 days on feed (DOF), and (4) after 135 DOF. DNA was extracted from feces for detection of virulence and serogroup genes by polymerase chain reaction (PCR) and immunomagnetic separation and pulsed-field gel electrophoresis (PFGE) were performed to collect and subtype STEC isolates...
October 25, 2016: Foodborne Pathogens and Disease
H Bayram, A Sayadi, J Goenaga, E Immonen, G Arnqvist
The seed beetle Callosobruchus maculatus is a significant agricultural pest and increasingly studied model of sexual conflict. Males possess genital spines that increase the transfer of seminal fluid proteins (SFPs) into the female body. As SFPs alter female behaviour and physiology, they are likely to modulate reproduction and sexual conflict in this species. Here, we identified SFPs using proteomics combined with a de novo transcriptome. A prior 2D-sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis identified male accessory gland protein spots that were probably transferred to the female at mating...
October 24, 2016: Insect Molecular Biology
D-X Ban, Y Liu, T-W Cao, S-J Gao, S-Q Feng
STUDY DESIGN: The rat's acellular spinal cord scaffold (ASCS) and spinal cord neurons were prepared in vitro to explore their biocompatibility. OBJECTIVES: The preparation of ASCS and co-culture with neuron may lay a foundation for clinical treatment of spinal cord injury (SCI). SETTING: Tianjin Medical University General Hospital, ChinaMethods:ASCS was prepared by chemical extraction method. Hematoxylin and eosin (H&E), myelin staining and scanning electron microscope were used to observe the surface structure of ASCS...
October 25, 2016: Spinal Cord
Dillon Hawley, Hema Aluri, Helene Armaos, Gina Kim, Claire Kublin, Driss Zoukhri
PURPOSE: Gene expression and protein analysis studies require high-quality human tissue which is a challenge and difficult to obtain through live human biopsies. Human postmortem lacrimal gland (LG) and submandibular gland (SMG) tissues have the potential to provide an invaluable source for studying the mechanisms involved in LG and SMG dysfunction. Therefore, we aimed to test the suitability of post-mortem LG and SMG for molecular, biochemical, and cell biological studies. METHODS: LG and SMG tissue from healthy donors was collected and immediately placed in RNAlater solution and then shipped overnight at 4 °C...
2016: Molecular Vision
Jasmeet Singh, Imran Sheikh, Prachi Sharma, Satish Kumar, Shailender Kumar Verma, Rahul Kumar, Priyanka Mathpal, Sundip Kumar, Pritesh Vyas, H S Dhaliwal
High molecular weight glutenin subunits (HMWGS) are responsible for dough elasticity and bread making quality of bread wheat. Related wild non-progenitor species, Aegilops kotschyi possesses higher molecular weight x and y glutenin subunits than the bread wheat cultivars. A wheat-Aegilops substitution line with 1U chromosome was used for the transfer of (HMWGS) of 1U to wheat by using pollen radiation hybridization approach. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis profiling showed different patterns of allelic variations with either the presence or absence of HMWGS, Glu-1A (1, null), Glu-1B (7, 7 + 8, 17 + 18) and Glu-1D (5 + 10, 2 + 12, null)...
September 2016: Journal of Food Science and Technology
M A Malik, H K Sharma, C S Saini
The present study evaluated the effect of removal of polyphenols on the structural properties of protein isolates extracted from sunflower seed and kernel. The structural and thermal changes in protein upon phenolic interaction were studied using circular dichroism, differential scanning calorimetry, thermal gravimetric analysis, X-ray diffraction, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and Fourier Transform Infrared (FT-IR) spectroscopy. Presence of phenolic compounds in proteins decreased the ordered structure content with parallel increase in unordered structure content...
September 2016: Journal of Food Science and Technology
Graziano Colombo, Marco Clerici, Alessandra Altomare, Francesco Rusconi, Daniela Giustarini, Nicola Portinaro, Maria Lisa Garavaglia, Ranieri Rossi, Isabella Dalle-Donne, Aldo Milzani
: In this study, we assessed the oxidative damage occurring in plasma proteins when human blood was exposed to inflammatory concentrations of hypochlorous acid (HOCl). We used specific thiol labelling and Western blot analyses to determine protein thiol oxidation, as well as analytical gel filtration HPLC coupled to fluorescence detection to explore formation of high molecular weight (HMW) protein aggregates. Thiol-containing proteins oxidized by HOCl were identified by redox proteomics...
October 21, 2016: Journal of Proteomics
Wipada Duangprompo, Kalaya Aree, Arunporn Itharat, Pintusorn Hansakul
5,6-dihydroxy-2,4-dimethoxy-9,10-dihydrophenanthrene (HMP) is an active compound isolated from the rhizome extracts of Dioscorea membranacea Pierre, a Thai medicinal plant. This study aimed to investigate the growth-inhibitory and apoptosis-inducing effects of HMP in human lung cancer A549 cells. The antiproliferative and cytotoxic effects of HMP were analyzed by a Sulforhodamine B assay. Cell division, cell cycle distribution and membrane asymmetry changes were each performed with different fluorescent dyes and then analyzed by flow cytometry...
October 25, 2016: American Journal of Chinese Medicine
C Patrick McAtee, Christopher A Seid, Molly Hammond, Elissa Hudspeth, Brian P Keegan, Zhuyun Liu, Junfei Wei, Bin Zhan, Raul Arjona-Sabido, Vladimir Cruz-Chan, Eric Dumonteil, Peter J Hotez, Maria Elena Bottazzi
The nucleoside hydrolase gene from Leishmania donovani was cloned and expressed in Escherichia coli as a full length 36-kDa protein (LdNH36). Following lysis and extraction, the protein was purified by anion exchange and gel filtration chromatography. The purified protein had a molecular mass of approximately 36-kDa and was confirmed to be >99% pure. Using a nucleoside hydrolase assay, the protein was found to exhibit a Km of 741 ± 246 μM. Protein integrity was confirmed by lithium dodecyl sulfate polyacrylamide gel electrophoresis (LDS-PAGE), mass spectrometry (MS), and enzymatic assay...
October 20, 2016: Protein Expression and Purification
Ceyda Acilan, Zelal Adiguzel, Buse Cevatemre, Didem Karakas, Engin Ulukaya, Nádia Ribeiro, Isabel Correia, João Costa Pessoa
BACKGROUND: To overcome the hurdles of cisplatin, majorly its toxicity and resistance, there has been extensive search for alternative anti-cancer metal-based compounds. Here, three Cu(II)-complexes, Cu(Sal-Gly)(phen), Cu(Sal-Gly)(pheamine), Cu(Sal-Gly)(phepoxy) are characterized for their interaction with DNA, cytotoxicity and mechanism of action. METHODS: The binding ability of the complexes to Calf-Thymus DNA was evaluated by competition fluorescence studies with thiazole-orange, UV-Vis and circular-dichroism spectroscopic titrations...
October 20, 2016: Biochimica et Biophysica Acta
Monica Dalva, Khadija El Jellas, Solrun J Steine, Bente B Johansson, Monika Ringdal, Janniche Torsvik, Heike Immervoll, Dag Hoem, Felix Laemmerhirt, Peter Simon, Markus M Lerch, Stefan Johansson, Pål R Njølstad, Frank U Weiss, Karianne Fjeld, Anders Molven
BACKGROUND/OBJECTIVES: We have recently described copy number variants (CNVs) of the human carboxyl-ester lipase (CEL) gene, including a recombined deletion allele (CEL-HYB) that is a genetic risk factor for chronic pancreatitis. Associations with pancreatic disease have also been reported for the variable number of tandem repeat (VNTR) region located in CEL exon 11. Here, we examined if CEL CNVs and VNTR length polymorphisms affect the risk for developing pancreatic cancer. METHODS: CEL CNVs and VNTR were genotyped in a German family with non-alcoholic chronic pancreatitis and pancreatic cancer, in 265 German and 197 Norwegian patients diagnosed with pancreatic adenocarcinoma, and in 882 controls...
October 11, 2016: Pancreatology: Official Journal of the International Association of Pancreatology (IAP) ... [et Al.]
Kang Liao, Yili Chen, Menghe Wang, Penghao Guo, Qiwen Yang, Yuxing Ni, Yunsong Yu, Bijie Hu, Ziyong Sun, Wenxiang Huang, Yong Wang, Anhua Wu, Xianju Feng, Yanping Luo, Zhidong Hu, Yunzhuo Chu, Shulan Chen, Bin Cao, Jianrong Su, Bingdong Gui, Qiong Duan, Shufang Zhang, Haifeng Shao, Haishen Kong, Yingchun Xu
BACKGROUND: Recently, the emergence of multidrug-resistant organisms such as extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae has raised considerable concern regarding the appropriate treatment of intra-abdominal infections (IAIs). In this study, we investigated the molecular characteristics of ESBL among clinical isolates of Escherichia coli and Klebsiella pneumoniae causing IAIs and their pattern of antimicrobial resistance, which can provide useful information about the epidemiology and risk factors associated with these infections...
October 7, 2016: Diagnostic Microbiology and Infectious Disease
Mariantonietta Succi, Maria Aponte, Patrizio Tremonte, Serena Niro, Elena Sorrentino, Massimo Iorizzo, Luca Tipaldi, Gianfranco Pannella, Gianfranco Panfili, Alessandra Fratianni, Raffaele Coppola
Five protected designation of origin (PDO) Caciocavallo Silano and 6 non-PDO Caciocavallo cheeses, ripened for 6 mo and collected in the 5 Italian regions of the PDO area (Apulia, Basilicata, Calabria, Campania, and Molise, Italy), were studied to assess their physico-chemical (pH, acidity, moisture, fat, ash, protein, and free amino acid composition) and microbiological profiles. Analyses evidenced a certain fluctuation of previous parameters among samples regardless of the kind of cheese evaluated (PDO and non-PDO)...
October 19, 2016: Journal of Dairy Science
Roel Haesendonck, Geertrui Rasschaert, An Martel, Elin Verbrugghe, Marc Heyndrickx, Freddy Haesebrouck, Frank Pasmans
Salmonella enterica infections in pigeons are generally associated with pigeon-adapted strains of serovar Typhimurium that are of little public health concern. Here, we isolated Salmonella Enteritidis phage type 4 (PT4), an important human pathogen, from a population of feral pigeons in Brussels, which was further characterized by Pulsed-Field Gel Electrophoresis. All pigeon isolates belonged to the same pulsotype, which has been present in Belgian pigeons at least since 2001 and is associated with poultry and disease in humans...
November 15, 2016: Veterinary Microbiology
Ganesan Kumaravel, Natarajan Raman
Two novel imidazole derived Schiff bases, (Z)-1-(1H-benzo[d]imidazol-2-yl)-N-benzylidenemethanamine (L(1)) and 1-(1H-benzo[d]imidazol-2-yl)-N-(4-nitrobenzylidene) methanamine, and a series of their transition metal complexes of the types [M(L(1))2]Cl2 and [M(L(2))2]Cl2 where, M=Cu(II), Ni(II), Co(II) and Zn(II) have been designed and synthesized. These compounds were characterized by various spectral and physicochemical data. UV-Vis, magnetic susceptibility and molar conductivity data indicate that all the complexes adopt square planar geometry...
January 1, 2017: Materials Science & Engineering. C, Materials for Biological Applications
Lin Li, Kaixiong Yang, Jian Lin, Junxin Liu
An integrated-bioreactor, which consisted of a suspended zone and an immobilized zone, was applied to treat gases containing SO2. The removal of SO2 in suspended zone differed slightly from that in immobilized zone. The influences of operational aspects such as SO2 load, temperature, and pH on integrated-bioreactor performance and bacterial community composition were investigated. The synergistic action of the two zones led to effective reduction of SO2, and the total removal efficiencies with the inlet concentration of 91-117 mg/m(3), were over 85 % in steady state...
October 21, 2016: Bioprocess and Biosystems Engineering
Katsutoshi Yamasaki, Masaru Komatsu, Tamotsu Ono, Hisaaki Nishio, Noriyuki Sueyoshi, Kaneyuki Kida, Kaori Satoh, Hirofumi Toda, Isao Nishi, Masahiro Akagi, Tetsu Mizutani, Isako Nakai, Tomomi Kofuku, Tamaki Orita, Takuya Zikimoto, Tatsuya Nakamura, Yasunao Wada
Six Klebsiella pneumoniae clinical isolates resistant to various cephalosporins and cephamycins were identified in a Japanese general hospital, a tertiary care hospital, between November 2009 and April 2010. All K. pneumoniae isolates carried blaGES-4 and blaSHV-1, while 2 K. pneumoniae isolates also harbored blaCTX-M-15. The pulsed-field gel electrophoresis patterns revealed that these 6 K. pneumoniae isolates were almost identical, suggesting their clonal relatedness. Plasmid profiles and conjugation assays revealed that these blaGES-4 genes were located on similar conjugative plasmids...
October 18, 2016: Journal of Infection and Chemotherapy: Official Journal of the Japan Society of Chemotherapy
Gloria Isani, Enea Ferlizza, Aurora Cuoghi, Elisa Bellei, Emanuela Monari, Barbara Bianchin Butina, Carolina Castagnetti
Characterisation of the physiologic equine amniotic fluid (AF) proteome is a prerequisite to study its changes during diseases and discover new biomarkers. The aim of this study was to identify by a proteomic approach the most abundant proteins of equine AF. AF samples were collected at parturition from 24 healthy mares that delivered healthy foals. All samples were subjected to sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) on 4-12% gels. A pool of the 24 samples, after SDS-PAGE, was cut in 25 slices, trypsin-digested and analysed by mass spectrometry (MS) for protein identification...
October 11, 2016: Animal Reproduction Science
Xiuli Chen, Hao Chen, Chunyan Lu, Chao Yang, Xiaoqi Yu, Kun Li, Yongmei Xie
In this report, the design of two novel nitrogen mustard-based DNA cross-linking agents with fluorophores incorporated into the structure (TN-A and TN-B) is disclosed. The results indicate that TN-A and TN-B can serve directly as both reporting and imaging agents for flow cytometry and gel electrophoresis without the necessity of another fluorescent tagging agents. TN-A and TN-B both selectively locate in the mitochondria and exhibit good antitumor activity. Notably, TN-A is the first DNA crosslinking agent with near infrared fluorescence emission properties...
December 1, 2016: Talanta
Lukasz P Kozlowski
BACKGROUND: Accurate estimation of the isoelectric point (pI) based on the amino acid sequence is useful for many analytical biochemistry and proteomics techniques such as 2-D polyacrylamide gel electrophoresis, or capillary isoelectric focusing used in combination with high-throughput mass spectrometry. Additionally, pI estimation can be helpful during protein crystallization trials. RESULTS: Here, I present the Isoelectric Point Calculator (IPC), a web service and a standalone program for the accurate estimation of protein and peptide pI using different sets of dissociation constant (pKa) values, including two new computationally optimized pKa sets...
October 21, 2016: Biology Direct
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