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https://www.readbyqxmd.com/read/28213262/one-single-fast-and-robust-capillary-electrophoresis-method-for-the-direct-quantification-of-intact-adenovirus-particles-in-upstream-and-downstream-processing-samples
#1
Ewoud van Tricht, Lars Geurink, Harold Backus, Marta Germano, Govert W Somsen, Cari E Sänger-van de Griend
During development of adenovirus-based vaccines, samples have to be analyzed in order to either monitor the production process or control the quality and safety of the product. An important quality attribute is the total concentration of intact adenoviruses, which currently is determined by quantitative polymerase chain reaction (qPCR) or anion exchange-HPLC. Capillary Electrophoresis (CE) was evaluated as alternative to the current methods with the aim to have one single method that allows reliable and fast quantification of adenovirus particles throughout the full process...
May 1, 2017: Talanta
https://www.readbyqxmd.com/read/28213091/convergence-of-hepcidin-deficiency-systemic-iron-overloading-heme-accumulation-and-rev-erb%C3%AE-%C3%AE-activation-in-aryl-hydrocarbon-receptor-elicited-hepatotoxicity
#2
Kelly A Fader, Rance Nault, Mathew P Kirby, Gena Markous, Jason Matthews, Timothy R Zacharewski
Persistent aryl hydrocarbon receptor (AhR) agonists elicit dose-dependent hepatic lipid accumulation, oxidative stress, inflammation, and fibrosis in mice. Iron (Fe) promotes AhR-mediated oxidative stress by catalyzing reactive oxygen species (ROS) production. To further characterize the role of Fe in AhR-mediated hepatotoxicity, male C57BL/6 mice were orally gavaged with sesame oil vehicle or 0.01-30μg/kg 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) every 4days for 28days. Duodenal epithelial and hepatic RNA-Seq data were integrated with hepatic AhR ChIP-Seq, capillary electrophoresis protein measurements, and clinical chemistry analyses...
February 14, 2017: Toxicology and Applied Pharmacology
https://www.readbyqxmd.com/read/28210224/skin-mucus-of-gilthead-sea-bream-sparus-aurata-l-protein-mapping-and-regulation-in-chronically-stressed-fish
#3
Jaume Pérez-Sánchez, Genciana Terova, Paula Simó-Mirabet, Simona Rimoldi, Ole Folkedal, Josep A Calduch-Giner, Rolf E Olsen, Ariadna Sitjà-Bobadilla
The skin mucus of gilthead sea bream was mapped by one-dimensional gel electrophoresis followed by liquid chromatography coupled to high resolution mass spectrometry using a quadrupole time-of-flight mass analyzer. More than 2,000 proteins were identified with a protein score filter of 30. The identified proteins were represented in 418 canonical pathways of the Ingenuity Pathway software. After filtering by canonical pathway overlapping, the retained proteins were clustered in three groups. The mitochondrial cluster contained 59 proteins related to oxidative phosphorylation and mitochondrial dysfunction...
2017: Frontiers in Physiology
https://www.readbyqxmd.com/read/28208846/a-rare-case-of-multiple-myeloma-with-biclonal-gammopathy
#4
Abhik Banerjee, Kshama Pimpalgaonkar, Alap Lukiyas Christy
Multiple myeloma is a debilitating malignancy arising from plasma cells. These malignant plasma cells called myeloma cells proliferate and infiltrate the bone marrow. The disease is characterized by the presence of a monoclonal protein in plasma and/or the urine. In this report, we present a case of biclonal multiple myeloma which showed two M bands on serum protein electrophoresis. The patient had elevated serum IgA and IgG levels. To reveal the nature of M bands or clonality, serum Immunofixation study was performed which revealed IgA with Lambda and IgG with Kappa light chains...
December 2016: Journal of Clinical and Diagnostic Research: JCDR
https://www.readbyqxmd.com/read/28208654/a-protein-isolate-from-moringa-oleifera-leaves-has-hypoglycemic-and-antioxidant-effects-in-alloxan-induced-diabetic-mice
#5
Paulo C Paula, Daniele O B Sousa, Jose T A Oliveira, Ana F U Carvalho, Bella G T Alves, Mirella L Pereira, Davi F Farias, Martonio P Viana, Flavia A Santos, Talita C Morais, Ilka M Vasconcelos
Moringa oleifera has been used in traditional medicine to treat diabetes. However, few studies have been conducted to relate its antidiabetic properties to proteins. In this study, a leaf protein isolate was obtained from M. oleifera leaves, named Mo-LPI, and the hypoglycemic and antioxidant effects on alloxan-induced diabetic mice were assessed. Mo-LPI was obtained by aqueous extraction, ammonium sulphate precipitation and dialysis. The electrophoresis profile and proteolytic hydrolysis confirmed its protein nature...
February 11, 2017: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
https://www.readbyqxmd.com/read/28208145/carfilzomib-inhibits-constitutive-nf-%C3%AE%C2%BAb-activation-in-mantle-cell-lymphoma-b-cells-and-leads-to-the-induction-of-apoptosis
#6
Yong-Li Zhang, Matthew Ho Zhi Guang, Hui-Qin Zhuo, Xiang-Hui Min, Qin Yao, Ai-Qi Gu, Sheng-Hua Wu, Dai-Bo Zhang, Jing-Yuan Lu, Yan Chen, Yu-Han Chen, Ke-Jie Zhang
Mantle cell lymphoma (MCL) remains incurable and new treatments are needed, especially in the relapsed/refractory setting. We therefore investigated the effects of carfilzomib, a novel, long-acting, second-generation proteasome inhibitor, in MCL cells. Eight established MCL cell lines and freshly isolated primary MCL cells were treated with carfilzomib. Cell proliferation was assessed by a 3H-thymidine incorporation assay. Cell apoptosis was evaluated by flow cytometry with annexin V and propidium iodide. Electrophoresis mobility shift (EMSA), Western blot, and luciferase assays were used to analyze NF-κB activation and related signaling proteins...
February 17, 2017: Acta Haematologica
https://www.readbyqxmd.com/read/28207866/a-novel-mechanism-of-metal-gel-shift-by-histidine-rich-ni2-binding-hpn-protein-from-helicobacter-pylori-strain-ss1
#7
Rahul Mahadev Shelake, Yuki Ito, Junya Masumoto, Eugene Hayato Morita, Hidenori Hayashi
Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) is a universally used method for determining approximate molecular weight (MW) in protein research. Migration of protein that does not correlate with formula MW, termed "gel shifting" appears to be common for histidine-rich proteins but not yet studied in detail. We investigated "gel shifting" in Ni2+-binding histidine-rich Hpn protein cloned from Helicobacter pylori strain SS1. Our data demonstrate two important factors determining "gel shifting" of Hpn, polyacrylamide-gel concentration and metal binding...
2017: PloS One
https://www.readbyqxmd.com/read/28207001/genome-editing-using-facs-enrichment-of-nuclease-expressing-cells-and-indel-detection-by-amplicon-analysis
#8
Lindsey A Lonowski, Yoshiki Narimatsu, Anjum Riaz, Catherine E Delay, Zhang Yang, Francesco Niola, Katarzyna Duda, Elke A Ober, Henrik Clausen, Hans H Wandall, Steen H Hansen, Eric P Bennett, Morten Frödin
This protocol describes methods for increasing and evaluating the efficiency of genome editing based on the CRISPR-Cas9 (clustered regularly interspaced short palindromic repeats-CRISPR-associated 9) system, transcription activator-like effector nucleases (TALENs) or zinc-finger nucleases (ZFNs). First, Indel Detection by Amplicon Analysis (IDAA) determines the size and frequency of insertions and deletions elicited by nucleases in cells, tissues or embryos through analysis of fluorophore-labeled PCR amplicons covering the nuclease target site by capillary electrophoresis in a sequenator...
March 2017: Nature Protocols
https://www.readbyqxmd.com/read/28204975/surface-and-exoproteomes-of-gram-positive-pathogens-for-vaccine-discovery
#9
Massimiliano Biagini, Fabio Bagnoli, Nathalie Norais
Reverse vaccinology has been very successful in the discovery of vaccine candidates against many pathogenic bacteria by integrating genome and proteome mining. This great achievement was facilitated by the complementarity of the in silico prediction of antigens and the empirical data on protein localization, expression, and immunogenicity obtained through different techniques based on electrophoresis, immunoblotting and mass spectrometry. An iterative process between information provided by DNA sequence analysis and proteomic data has been established leading to precise antigen identification...
February 16, 2017: Current Topics in Microbiology and Immunology
https://www.readbyqxmd.com/read/28198502/extraction-of-proteins-and-preliminary-characterization-of-physicochemical-properties-in-toona-sinensis-fruit
#10
W-Z Li, W-N Han, B Liu, S-H Ding, X-K Zhang, R-S Wang
We investigated the extraction of Toona sinensis fruit proteins and preliminarily characterized their physicochemical properties. The results showed that optimal extraction occurred under conditions of pH 10.5, a duration of 40 min, a liquid-to-solid ratio of 25:1, and a temperature of 40°C by an orthogonal design using T. sinensis fruit protein as the index and single factor. The total nitrogen content was 13.8 g/100 g and included 17 different amino acids. The glutamate level was highest at 35.37%, followed by arginine at 15...
February 8, 2017: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/28197596/amphiphilic-polymers-formed-from-ring-opening-polymerization-a-strategy-for-the-enhancement-of-gene-delivery
#11
Yi-Mei Zhang, Zheng Huang, Ji Zhang, Wan-Xia Wu, Yan-Hong Liu, Xiao-Qi Yu
Cationic liposomes and polymers are both important candidates for use as non-viral gene vectors. However, both of them have special shortcomings and application limits. This work is devoted to the combination of advantages of liposomes and polymers. The ring-opening polymerization strategy was used for the preparation of amphiphilic polymers from cyclen-based cationic small lipids. The non-hydrophobic polymer and the corresponding lipids were also prepared for performing structure-activity relationship studies...
February 15, 2017: Biomaterials Science
https://www.readbyqxmd.com/read/28197578/chemical-characterization-of-the-glycated-myofibrillar-proteins-from-grass-carp-ctenopharyngodon-idella-and-their-impacts-on-the-human-gut-microbiota-in-vitro-fermentation
#12
Kaining Han, Ye Yao, Shiyuan Dong, Sun Jin, Hang Xiao, Haohao Wu, Mingyong Zeng
In this study, the chemical characterization of glycoconjugates of myofibrillar proteins from grass carp conjugated with glucose via Maillard reaction for up to 24 h of dry-heating was investigated, and their impacts on the microbial community in vitro human fecal fermentation were firstly evaluated by high-throughput sequencing technologies. The glycation greatly increased the furosine levels in glycoconjugates, which reached the maximum level (2.87 ± 0.08 mg per 100 mg protein) for 9 h of heating, and resulted in the structural changes of myofibrillar proteins based on the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Fourier transform infrared spectroscopy (FTIR) analysis...
February 15, 2017: Food & Function
https://www.readbyqxmd.com/read/28196296/age-related-reduction-of-dermal-fibroblast-size-up-regulates-multiple-matrix-metalloproteinases-as-observed-in-aged-human-skin-in-vivo
#13
Z Qin, R M Balimunkwe, T Quan
BACKGROUND: Fragmentation of collagen fibrils, the major structure protein in skin, is a hallmark of dermal aging. Matrix metalloproteinases (MMPs) are largely responsible for fragmentation of collagen fibrils. However, the alteration of all known mammalian MMPs and the mechanism underlying altered expression of MMPs in chronologically aged human skin are less understood. OBJECTIVES: To quantify gene expression of all 23 known mammalian MMPs in sun-protected young and aged human skin in vivo, and investigate the potential mechanism underlying age-related alteration of multiple MMPs...
February 14, 2017: British Journal of Dermatology
https://www.readbyqxmd.com/read/28196092/characterization-of-cereal-%C3%AE-glucan-extracts-from-oat-and-barley-and-quantification-of-proteinaceous-matter
#14
Claudia Zielke, Ondrej Kosik, Marie-Louise Ainalem, Alison Lovegrove, Anna Stradner, Lars Nilsson
An extraction method for mixed-linkage β-glucan from oat and barley was developed in order to minimize the effect of extraction on the β-glucan structure. β-Glucan were characterized in terms of molecular size and molar mass distributions using asymmetric flow field-flow fractionation (AF4) coupled to multiangle light scattering (MALS), differential refractive index (dRI) and fluorescence (FL) detection. The carbohydrate composition of the extracts was analysed using polysaccharide analysis by carbohydrate gel electrophoresis (PACE) and high-performance anion-exchange chromatography (HPAEC)...
2017: PloS One
https://www.readbyqxmd.com/read/28191937/a-versatile-platform-for-the-synthesis-of-orthogonally-cleavable-heteromultifunctional-crosslinkers
#15
Michelle R Sorkin, Joshua A Walker, Joseph S Brown, Christopher A Alabi
Cleavable and heteromultifunctional crosslinkers have proven critical in a wide range of biological applications. Traditional approaches for synthesizing these linkers suffer from various synthetic and functional limitations. In this work, an efficient sequence-defined synthetic methodology, developed for the assembly of oligothioetheramides, was used to address many of these limitations. Four heterotrifunctional crosslinkers with up to two orthogonal internal cleavage sites were synthesized. These linkers were conjugated to a pair of fluorophores that undergo Förster resonance energy transfer (FRET) and a model protein - human transferrin...
February 13, 2017: Bioconjugate Chemistry
https://www.readbyqxmd.com/read/28191477/incorporation-of-%C3%AE-actin-loading-control-into-zymography
#16
Natasha Govindasamy, MengJie Yan, Paul Jurasz
Gelatin zymography and immunoblot are widely used gel electrophoresis techniques to study matrix metalloproteinases-2 and -9. Each method has its advantages and disadvantages. Zymography is exquisitely sensitive but offers no loading control to ensure equal sample loading. Immunoblot is a 100-1000-fold less sensitive, but allows for the probing of a sample loading control such as β-actin to ensure accurate protein loading. In this report, we describe two simple protocols that combine gelatin zymography to study MMP-2 and -9 levels with an in-gel β-actin immunoblot loading control, thus combining sensitivity and accuracy in a single assay...
November 1, 2016: Journal of Biological Methods
https://www.readbyqxmd.com/read/28190743/a-medically-relevant-capsular-polysaccharide-in-acinetobacter-baumannii-is-a-potential-vaccine-candidate
#17
Feng-Ling Yang, Tze-Chi Lou, Shu-Chen Kuo, Wan-Ling Wu, Jeffy Chern, Yi-Tzu Lee, Shui-Tsung Chen, Wei Zou, Nien-Tsung Lin, Shih-Hsiung Wu
Concerns of Acinetobacter baumannii infection have increased due to the emergence of multi-drug resistance. In the present study, we determined the capsular polysaccharide (CPS) structure of A. baumannii SK44, a clinical isolate from Taiwan, to consist of pentasaccharide repeats. We found that CPS-induced antibody provided 55% protection against challenge in an animal model. The CPS-specific antibody reacted with the surface components of about 62% clinical isolates (342/554 strains) from cross-sectional and longitudinal studies by dot-immunoassay...
February 9, 2017: Vaccine
https://www.readbyqxmd.com/read/28188804/metabolite-profiling-of-infection-associated-metabolic-markers-of-onchocerciasis
#18
Sasisekhar Bennuru, Sara Lustigman, David Abraham, Thomas B Nutman
The global efforts for onchocerciasis elimination may require additional tools (safe micro and macrofilaricidal drugs, vaccines and biomarkers) as elimination efforts move toward the "end game". Efforts toward the identification of suitable biomarkers have focused on specific protein(s) and/or nucleic acids, but metabolites present an alternative option as they have limited half-lives and are the result of combinatorial effects. In comparison to previously used methodology of LC-MS for metabolomic approaches, we used a non-targeted capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) to analyze the serum metabolic profiles of Ov-infected and -uninfected individuals (n=20)...
February 8, 2017: Molecular and Biochemical Parasitology
https://www.readbyqxmd.com/read/28187683/proteomics-profiling-of-chimeric-truncated-tissue-plasminogen-activator-producing-chinese-hamster-ovary-cells-cultivated-in-a-chemically-defined-medium-supplemented-with-protein-hydrolysates
#19
Bahareh Azarian, Seyedeh Matin Sajedin, Amin Azimi, Mozhgan Raigani, Behrouz Vaziri, Fatemeh Davami
Background: Culture media enrichment through the addition of protein hydrolysates is beneficial for achieving higher protein expression. Methods: In this study, designing the optimum mixture of four soy and casein-derived hydrolysates was successfully performed by design of experiment and specific productivity increased in all predicted combinations. Protein profile of recombinant CHO (rCHO) cells producing tissue plasminogen activator in a serum-free medium (SFM) supplemented with designed hydrolysate additives was compared to that of rCHO cells cultivated in SFM...
11, 2017: Iranian Biomedical Journal
https://www.readbyqxmd.com/read/28187457/the-b-box-module-of-cyld-is-responsible-for-its-intermolecular-interaction-and-cytoplasmic-localization
#20
Songbo Xie, Miao Chen, Siqi Gao, Tao Zhong, Peng Zhou, Dengwen Li, Jun Zhou, Jinmin Gao, Min Liu
The tumor suppressor protein cylindromatosis (CYLD), as a microtubule-associated deubiquitinase, plays a pivotal role in a wide range of cellular activities, including innate immunity, cell division, and ciliogenesis. Structural characterization reveals a small zinc-binding B-box inserted within the ubiquitin specific protease (USP) domain of CYLD; however, the exact role for this module remains yet to be elucidated. Here we identify a critical role for the B-box in facilitating the intermolecular interaction and subcellular localization of CYLD...
February 7, 2017: Oncotarget
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