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https://www.readbyqxmd.com/read/29674116/a-versatile-expression-vector-for-the-growth-and-amplification-of-unmodified-phage-display-polypeptides
#1
Alexander J Winton, Janae L Baptiste, Mark A Allen
Proteins and polypeptides represent nature's most complex and versatile polymer. They provide complicated shapes, diverse chemical functionalities, and tightly regulated and controlled sizes. Several disease states are related to the misfolding or overproduction of polypeptides and yet polypeptides are present in several therapeutic molecules. In addition to biological roles; short chain polypeptides have been shown to interact with and drive the bio-inspired synthesis or modification of inorganic materials...
April 16, 2018: Protein Expression and Purification
https://www.readbyqxmd.com/read/29673162/nanoscale-zinc-oxide-particles-for-improving-the-physiological-and-sanitary-quality-of-a-mexican-landrace-of-red-maize
#2
Juan Estrada-Urbina, Alejandro Cruz-Alonso, Martha Santander-González, Abraham Méndez-Albores, Alma Vázquez-Durán
In this research, quasi-spherical-shaped zinc oxide nanoparticles (ZnO NPs) were synthesized by a simple cost-competitive aqueous precipitation method. The engineered NPs were characterized using several validation methodologies: UV–Vis spectroscopy, diffuse reflection UV–Vis, spectrofluorometry, transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA), and Fourier transform infrared (FTIR) spectroscopy with attenuated total reflection (ATR). A procedure was established to coat a landrace of red maize using gelatinized maize starch...
April 17, 2018: Nanomaterials
https://www.readbyqxmd.com/read/29669465/superiority-of-rat-over-murine-model-for-studies-on-the-evolution-of-cancer-genome
#3
Shinya Akatsuka, Guang Hua Li, Shinya Toyokuni
Evolution of the species and carcinogenesis are similar in that genomic alterations are the key events. Oxidative stress derived from various etiologies is one of the major causes of carcinogenesis by inducing mutations in the genome. Persistent oxidative stress in the renal proximal tubules through Fenton reaction catalyzed by ferric nitrilotriacetate (Fe-NTA) generates renal cell carcinoma (RCC) in mice and rats. Here, in order to observe the species difference in oxidative stress-induced carcinogenesis and to obtain an insight regarding the characteristics of each species, we compared the genomic alterations using array-based comparative genome hybridization among RCCs in Mutyh knockout/wild-type mice (C57BL/6 background) induced by Fe-NTA, RCCs in F1 hybrids of Brown-Norway/Fischer-344 wild-type rats and clear cell renal cell carcinoma (CCRCC)/papillary renal cell carcinoma (PRCC) of humans...
April 18, 2018: Free Radical Research
https://www.readbyqxmd.com/read/29663061/sizing-lipid-droplets-from-adult-and-geriatric-mouse-liver-tissue-via-nanoparticle-tracking-analysis
#4
Katherine A Muratore, Charles P Najt, Nicholas M Livezey, James Marti, Douglas G Mashek, Edgar A Arriaga
The significance of lipid droplets in lipid metabolism, cell signaling, and regulating longevity is increasingly recognized, yet the lipid droplet's unique properties and architecture make it difficult to size and study using conventional methods. To begin to address this issue, we demonstrate the capabilities of nanoparticle tracking analysis (NTA) for sizing of lipid droplets. NTA was found to be adequate to assess lipid droplet stability over time, indicating that lipid droplet preparations are stable for up to 24 h...
April 17, 2018: Analytical and Bioanalytical Chemistry
https://www.readbyqxmd.com/read/29660899/plasma-exosome-levels-in-non-small-cell-lung-cancer-correlation-with-clinicopathological-features-and-prognostic-implications
#5
Qingyun Liu, Ying Xiang, Shuai Yuan, Weijia Xie, Chengying Li, Zeyao Hu, Ying Xiang, Na Wu, Long Wu, Zubin Yu, Li Bai, Yafei Li
BACKGROUND: Biomarker studies revealed important clinical significance of exosome for cancer patients. However, there is currently no consensus on exosome quantification methods. METHODS: Bicinchoninic acid (BCA) method, acetylcholinesterase (AChE) method and nanoparticle tracking analysis (NTA) were utilized to quantify 20 plasma exosome samples, and interrelations between these three methods were explored. Associations of plasma exosome levels with characteristics and prognosis of 208 non-small-cell lung cancer (NSCLC) patients were investigated...
April 3, 2018: Cancer Biomarkers: Section A of Disease Markers
https://www.readbyqxmd.com/read/29656860/truncating-variants-in-naa15-are-associated-with-variable-levels-of-intellectual-disability-autism-spectrum-disorder-and-congenital-anomalies
#6
Hanyin Cheng, Avinash V Dharmadhikari, Sylvia Varland, Ning Ma, Deepti Domingo, Robert Kleyner, Alan F Rope, Margaret Yoon, Asbjørg Stray-Pedersen, Jennifer E Posey, Sarah R Crews, Mohammad K Eldomery, Zeynep Coban Akdemir, Andrea M Lewis, Vernon R Sutton, Jill A Rosenfeld, Erin Conboy, Katherine Agre, Fan Xia, Magdalena Walkiewicz, Mauro Longoni, Frances A High, Marjon A van Slegtenhorst, Grazia M S Mancini, Candice R Finnila, Arie van Haeringen, Nicolette den Hollander, Claudia Ruivenkamp, Sakkubai Naidu, Sonal Mahida, Elizabeth E Palmer, Lucinda Murray, Derek Lim, Parul Jayakar, Michael J Parker, Stefania Giusto, Emanuela Stracuzzi, Corrado Romano, Jennifer S Beighley, Raphael A Bernier, Sébastien Küry, Mathilde Nizon, Mark A Corbett, Marie Shaw, Alison Gardner, Christopher Barnett, Ruth Armstrong, Karin S Kassahn, Anke Van Dijck, Geert Vandeweyer, Tjitske Kleefstra, Jolanda Schieving, Marjolijn J Jongmans, Bert B A de Vries, Rolph Pfundt, Bronwyn Kerr, Samantha K Rojas, Kym M Boycott, Richard Person, Rebecca Willaert, Evan E Eichler, R Frank Kooy, Yaping Yang, Joseph C Wu, James R Lupski, Thomas Arnesen, Gregory M Cooper, Wendy K Chung, Jozef Gecz, Holly A F Stessman, Linyan Meng, Gholson J Lyon
N-alpha-acetylation is a common co-translational protein modification that is essential for normal cell function in humans. We previously identified the genetic basis of an X-linked infantile lethal Mendelian disorder involving a c.109T>C (p.Ser37Pro) missense variant in NAA10, which encodes the catalytic subunit of the N-terminal acetyltransferase A (NatA) complex. The auxiliary subunit of the NatA complex, NAA15, is the dimeric binding partner for NAA10. Through a genotype-first approach with whole-exome or genome sequencing (WES/WGS) and targeted sequencing analysis, we identified and phenotypically characterized 38 individuals from 33 unrelated families with 25 different de novo or inherited, dominantly acting likely gene disrupting (LGD) variants in NAA15...
April 9, 2018: American Journal of Human Genetics
https://www.readbyqxmd.com/read/29624018/development-of-a-rapid-immunochromatographic-assay-for-detection-of-antibodies-against-porcine-epidemic-diarrhea-virus
#7
R Li, X Tian, Y Yu, S Qiao, Z Wang, J Ma
An immunochromatographic test strip was developed for the detection of antibodies against porcine epidemic diarrhea virus (PEDV) in porcine serum. In this test, the recombinant nucleocapsid protein of PEDV was expressed in soluble form and purified by using Ni-NTA resin and gel filtration chromatography. The purified recombinant N protein conjugated with colloidal gold was dispensed on a conjugate pad as the detector. Staphylococcal protein A and rabbit anti-N protein IgG were blotted on a nitrocellulose membrane for the test and control lines, respectively...
March 2018: Polish Journal of Veterinary Sciences
https://www.readbyqxmd.com/read/29621784/exosomes-derived-from-dendritic-cells-attenuate-liver-injury-by-modulating-the-balance-of-treg-and-th17-cells-after-ischemia-reperfusion
#8
Lei Zheng, Zhi Li, Wei Ling, Deming Zhu, Zhiwen Feng, Lianbao Kong
BACKGROUND/AIMS: The present study aimed to evaluate the effects as well as the underlying mechanisms of bone marrow-derived dendritic cells (BMDCs) and exosomes produced by BMDCs (DEXs) on hepatic ischemia-reperfusion (I/R) injury (IRI). METHODS: Primary hepatocytes were isolated and used to mimic the liver IR microenvironment. BMDCs were induced and characterized both biochemically with a flow cytometer (FCM) and biophysically with a microscope. Then, we exposed BMDCs to the supernatants from primary hepatocytes and evaluated the maturation of BMDCs by FCM...
March 29, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29604370/the-application-of-step-technology%C3%A2-for-particle-and-protein-dispersion-detection-studies-in-biopharmaceutical-research
#9
J Gross, N Herrmann, M Blech, S R Pinnapireddy, P Garidel, U Bakowsky
Particle detection and analysis techniques are essential in biopharmaceutical industries to evaluate the quality of various parenteral formulations regarding product safety, product quality and to meet the regulations set by the authority agencies. Several particle analysis systems are available on the market, but for the operator, it is quite challenging to identify the suitable method to analyze the sample. At the same time these techniques are the basis to gain a better understanding in biophysical processes, e...
March 28, 2018: International Journal of Pharmaceutics
https://www.readbyqxmd.com/read/29596634/expression-and-purification-of-biologically-active-recombinant-rabbit-monocyte-chemoattractant-protein1-in-escherichia-coli
#10
Maryam Boshtam, Hossein Khanahmad Shahreza, Sadegh Feizollahzadeh, Ilnaz Rahimmanesh, Seddigheh Asgary
Monocyte chemoattractant protein 1 (MCP1) with recruiting monocytes is an important factor at the beginning of inflammatory disorders such as atherosclerosis which seems its blocking preclude this process and help improvement of related diseases. To perform clinical research in this field, MCP1 protein is required but firstly, animal studies should be done. As the rabbit is a suitable model for many inflammatory disorders, and Escherichia coli BL21(DE3) (BL21) cell is a high-efficiency host for protein expression, we decided to produce recombinant rabbit MCP1 (rRMCP1) in BL21/pET28a system...
March 27, 2018: FEMS Microbiology Letters
https://www.readbyqxmd.com/read/29593411/imaging-of-extracellular-vesicles-derived-from-human-bone-marrow-mesenchymal-stem-cells-using-fluorescent-and-magnetic-labels
#11
Sylwia Dabrowska, Andrea Del Fattore, Elzbieta Karnas, Malgorzata Frontczak-Baniewicz, Hanna Kozlowska, Maurizio Muraca, Miroslaw Janowski, Barbara Lukomska
Background: Mesenchymal stem cells have been shown therapeutic in various neurological disorders. Recent studies support the notion that the predominant mechanism by which MSCs act is through the release of extracellular vesicles (EVs). EVs seem to have similar therapeutic activity as their cellular counterparts and may represent an interesting alternative standalone therapy for various diseases. The aim of the study was to optimize the method of EV imaging to better understand therapeutic effects mediated by EVs...
2018: International Journal of Nanomedicine
https://www.readbyqxmd.com/read/29584404/dual-valve-and-counter-flow-surface-plasmon-resonance
#12
Xiaoying Wang, Feimeng Zhou
Two six-port injector valves and one selector valve commonly used in flow injection analysis are combined with a surface plasmon resonance instrument wherein solutions introduced from the two inlets counter-flow inside the flow cell. The system is rather versatile as the same or different solutions can be rapidly and repeatedly introduced to the two fluidic channels in series or in parallel. Unlike most commercial SPR instruments employing a single injector valve, solutions separately injected from the two injector valves can be readily exchanged (< 1 s) between the two channels...
March 27, 2018: Analytical Chemistry
https://www.readbyqxmd.com/read/29566124/sustained-release-of-endothelial-progenitor-cell-derived-extracellular-vesicles-from-shear-thinning-hydrogels-improves-angiogenesis-and-promotes-function-after-myocardial-infarction
#13
Carol W Chen, Leo L Wang, Samir Zaman, Jon Gordon, Maria F Arisi, Chantel M Venkataraman, Jennifer J Chung, George Hung, Ann C Gaffey, Lynn A Spruce, Hossein Fazelinia, Robert C Gorman, Steven H Seeholzer, Jason A Burdick, Pavan Atluri
Aims: Previous studies have demonstrated improved cardiac function following myocardial infarction (MI) after administration of endothelial progenitor cells (EPCs) into ischemic myocardium. A growing body of literature supports paracrine effectors, including extracellular vesicles (EVs), as the main mediators of the therapeutic benefits of EPCs. The use of paracrine factors is an attractive strategy that harnesses the effects of cell therapy without concerns of cell engraftment or viability...
March 16, 2018: Cardiovascular Research
https://www.readbyqxmd.com/read/29563556/near-native-site-specific-and-purification-free-protein-labeling-for-quantitative-protein-interaction-analysis-by-microscale-thermophoresis
#14
Tanja Bartoschik, Stefanie Galinec, Christian Kleusch, Katarzyna Walkiewicz, Dennis Breitsprecher, Sebastian Weigert, Yves A Muller, Changjiang You, Jacob Piehler, Thomas Vercruysse, Dirk Daelemans, Nuska Tschammer
MicroScale Thermophoresis (MST) is a frequently used method for the quantitative characterization of intermolecular interactions with several advantages over other technologies. One of these is its capability to determine equilibrium constants in solution including complex biological matrices such as cell lysates. MST requires one binding partner to be fluorescent, which is typically achieved by labeling target proteins with a suitable fluorophore. Here, we present a near-native, site-specific in situ labeling strategy for MST experiments that enables reliable measurements in cell lysates and that has distinct advantages over routine covalent labeling techniques...
March 21, 2018: Scientific Reports
https://www.readbyqxmd.com/read/29559329/molecular-characterization-of-babesia-microti-seroreactive-antigen-5-1-1-and-development-of-rapid-detection-methods-for-anti-b-microti-antibodies-in-serum
#15
Yu Chun Cai, Fen Wu, Wei Hu, Jiaxu Chen, Shao Hong Chen, Bin Xu, Yan Lu, Lin Ai, Chun Li Yang, Shimin Zhao
Babesiosis has become a new global threat impacting human health, and most human babesiosis cases are caused by Babesia microti. Until now few antigens of B. microti have been described which can be used for the diagnosis of human babesiosis. In the present study, we report on the bioinformatic analysis, cloning and expression of the sequence encoding the B. microti seroreactive antigen 5-1-1 to investigate its potential incorporation in serologic diagnostic tools for babesiosis. Bioinformatic analysis and recombinant gene expression were performed to molecularly characterize seroreactive antigen 5-1-1...
March 17, 2018: Acta Tropica
https://www.readbyqxmd.com/read/29557634/spatial-and-chemical-surface-guidance-of-nk-cell-cytotoxic-activity
#16
Guillaume Le Saux, Avishay Edri, Yossi Keydar, Uzi Hadad, Angel Porgador, Mark Schvartzman
Studying of how different signaling pathways spatially integrate in cells requires selective manipulation and control of different transmembrane ligand-receptor pairs at the same time. This work explores a novel method for precisely arranging two arbitrarily chosen ligands on a micron-scale 2D pattern. The approach is based on lithographic patterning of Au and TiO2 films, followed by their selective functionalization with Ni-NTA/histidine and biotin/avidin chemistries, respectively. The selectivity of chemical and biological functionalizations is demonstrated by XPS and immunofluorescence imaging, respectively...
March 20, 2018: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/29556723/biochemical-and-structural-characterization-of-a-novel-cold-active-esterase-like-protein-from-the-psychrophilic-yeast-glaciozyma-antarctica
#17
Noor Haza Fazlin Hashim, Nor Muhammad Mahadi, Rosli Md Illias, Shevin Rizal Feroz, Farah Diba Abu Bakar, Abdul Munir Abdul Murad
Dienelactone hydrolase, an α/β hydrolase enzyme, catalyzes the hydrolysis of dienelactone to maleylacetate, an intermediate for the Krebs cycle. Genome sequencing of the psychrophilic yeast, Glaciozyma antarctica predicted a putative open reading frame (ORF) for dienelactone hydrolase (GaDlh) with 52% sequence similarity to that from Coniophora puteana. Phylogenetic tree analysis showed that GaDlh is closely related to other reported dienelactone hydrolases, and distantly related to other α/β hydrolases...
March 20, 2018: Extremophiles: Life Under Extreme Conditions
https://www.readbyqxmd.com/read/29550816/sumoylation-of-ephb1-suppresses-neuroblastoma-tumorigenesis-via-inhibiting-pkc%C3%AE-activation
#18
Qin Chen, Rong Deng, Xian Zhao, Haihua Yuan, Hailong Zhang, Jinzhuo Dou, Ran Chen, Hui Jin, Yanli Wang, Jian Huang, Jianxiu Yu
BACKGROUND/AIMS: An increasing number of studies have linked <unterline>e</unterline>rythropoietin-<unterline>p</unterline>roducing <unterline>h</unterline>epatocellular carcinoma (Eph) family receptor tyrosine kinases to cancer progression. However, little knowledge is available about the regulation of their functions in cancer. METHODS: SUMOylation was analyzed by performing Ni2+-NTA pull-down assay and immunoprecipitation. Cell proliferation, anchorage-independent growth, and tumorigenesis in vivo were examined by cell counting kit-8, soft agar colony formation assay, and a xenograft tumor mouse model, respectively...
March 13, 2018: Cellular Physiology and Biochemistry
https://www.readbyqxmd.com/read/29545094/spherical-supported-membranes-as-platforms-for-screening-against-membrane-protein-targets
#19
V Vasilca, A Sadeghpour, S Rawson, L E Hawke, S A Baldwin, T Wilkinson, D Bannister, V L G Postis, M Rappolt, S P Muench, L J C Jeuken
Screening assays performed against membrane protein targets (e.g. phage display) are hampered by issues arising from protein expression and purification, protein stability in detergent solutions and epitope concealment by detergent micelles. Here, we have studied a fast and simple method to improve screening against membrane proteins: spherical-supported bilayer lipid membranes ("SSBLM"). SSBLMs can be quickly isolated via low-speed centrifugation and redispersed in liquid solutions while presenting the target protein in a native-like lipid environment...
March 12, 2018: Analytical Biochemistry
https://www.readbyqxmd.com/read/29542322/dynamic-scaling-of-exosome-sizes
#20
Michael E Paulaitis, Kitty Agarwal, Serge Patrick Nana-Sinkam
A model is proposed for characterizing exosome size distributions based on dynamic scaling of domain growth on the limiting membrane of multivesicular bodies in the established exosome biogenesis pathway. The scaling exponent in this model captures the asymmetry of exosome size distributions, which are notably right-skewed to larger vesicles independent of the minimum detectable vesicle size. Analyses of exosome size distributions obtained by cryo-TEM imaging and nanoparticle tracking (NTA) show, respectively, that the scaling exponent is sensitive to the state of the cell source for exosomes in cell culture supernatants, and can distinguish exosome size distributions in serum samples taken from cancer patients relative to those from healthy donors...
March 15, 2018: Langmuir: the ACS Journal of Surfaces and Colloids
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