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https://www.readbyqxmd.com/read/28076321/secretion-modification-region-derived-peptide-blocks-exosome-release-and-mediates-cell-cycle-arrest-in-breast-cancer-cells
#1
Ming-Bo Huang, Ruben R Gonzalez, James Lillard, Vincent C Bond
PURPOSE: Discovery and development of a novel anticancer PEG-SMR-Clu peptide to prevent breast cancer metastasis. How breast cancer cells and primary mammary epithelial cells interact and communicate with each other to promote tumorigenesis and how to prevent tumor metastasis has long been a concern of researchers. Cancer cells secrete exosomes containing proteins and RNA. These factors can influence tumor development by directly targeting cancer cells and tumor stroma. In this study, we determined the effects of a peptide as an inhibitor of exosome secretion on breast tumors...
January 5, 2017: Oncotarget
https://www.readbyqxmd.com/read/28058522/identification-and-characterization-of-a-novel-carboxylesterase-fpbh-that-hydrolyzes-aryloxyphenoxypropionate-herbicides
#2
Chenghong Wang, Jiguo Qiu, Youjian Yang, Jinwei Zheng, Jian He, Shunpeng Li
OBJECTIVE: To identify and characterize a novel aryloxyphenoxypropionate (AOPP) herbicide-hydrolyzing carboxylesterase from Aquamicrobium sp. FPB-1. RESULTS: A carboxylesterase gene, fpbH, was cloned from Aquamicrobium sp. FPB-1. The gene is 798 bp long and encodes a protein of 265 amino acids. FpbH is smaller than previously reported AOPP herbicide-hydrolyzing carboxylesterases and shares only 21-35% sequence identity with them. FpbH was expressed in Escherichia coli BL21(DE3) and the product was purified by Ni-NTA affinity chromatography...
January 5, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28054515/method-validation-of-nanoparticle-tracking-analysis-to-measure-pulmonary-nanoparticle-content-the-size-distribution-in-exhaled-breath-condensate-depends-on-occupational-exposure
#3
Jean-Jacques Sauvain, Guillaume Suarez, Jean-Louis Edmé, Olivia Maria de Paula Alvez Bezerra, Keller Silveira, Lenio Amaral, Ana Paula Carneiro, Natalie Cherot-Kornobis, Annie Sobaszek, Sébastien Hulo
A particle exposure assessment based on the dose deposited in the lungs would be the gold standard for the evaluation of any resulting health effects. Measuring particles in exhaled breath condensate (EBC)-a matrix containing water and airway lining fluid-could help to evaluate particle retention in the lungs. This study aimed to (1) validate a nanoparticle tracking analysis (NTA) method for determining the particle number concentration and their hydrodynamic size distribution in EBC, and (2) apply this method to EBC collected from workers exposed to soapstone (n = 55) or quartz dust (n = 12) and controls (n = 11)...
January 5, 2017: Journal of Breath Research
https://www.readbyqxmd.com/read/28044272/gene-cloning-and-expression-of-a-glucoside-3-dehydrogenase-from-sphingobacterium-faecium-zjf-d6-and-used-it-to-produce-n-p-nitrophenyl-3-ketovalidamine
#4
Jian-Fen Zhang, Wei-Qing Chen, Hong Chen
In this study, we report the cloning and expression of a functional glucoside 3-dehydrogenase (G3DH) gene from Sphingobacterium faecium ZJF-D6. This gene is 1686 bp in length and encodes a peptide of 562 amino acids. The G3DH gene was successfully expressed in E. coli, and the recombinant enzyme could oxidize glucosides, galactosides and analogues at C-3 position. The sequence and multiple alignment analysis showed that the enzyme has highest identity with G3DHs from Paraglaciecola polaris LMG 21857, Aliiglaciecola lipolytica E3 and Halomonas sp...
February 2017: World Journal of Microbiology & Biotechnology
https://www.readbyqxmd.com/read/28043510/effects-of-sorbitol-and-glycerol-on-the-structure-dynamics-and-stability-of-mycobacterium-tuberculosis-pyrazinamidase
#5
Mehrnoosh Khajehzadeh, Faramarz Mehrnejad, Mohammad Pazhang, Farahnoosh Doustdar
OBJECTIVE/BACKGROUND: Mycobacterium tuberculosis pyrazinamidase (PZase) is known an enzyme that is involved in degradation of pyrazinamide to ammonia and pyrazinoic acid. Pyrazinamide is an important first-line drug used in the short-course treatment of tuberculosis. Previous investigations have indicated that the pyrazinamide (PZA)-resistant M. tuberculosis strains are caused by point mutations in the PZase enzyme which is the activator of the prodrug PZA. Although the general fold of PZase was determined, the structural and functional properties of the enzyme in solution were not understood very well...
December 2016: International Journal of Mycobacteriology
https://www.readbyqxmd.com/read/28042333/nanosized-ucmsc-derived-extracellular-vesicles-but-not-conditioned-medium-exclusively-inhibit-the-inflammatory-response-of-stimulated-t-cells-implications-for-nanomedicine
#6
Marta Monguió-Tortajada, Santiago Roura, Carolina Gálvez-Montón, Josep Maria Pujal, Gemma Aran, Lucía Sanjurjo, Marcel la Franquesa, Maria-Rosa Sarrias, Antoni Bayes-Genis, Francesc E Borràs
Undesired immune responses have drastically hampered outcomes after allogeneic organ transplantation and cell therapy, and also lead to inflammatory diseases and autoimmunity. Umbilical cord mesenchymal stem cells (UCMSCs) have powerful regenerative and immunomodulatory potential, and their secreted extracellular vesicles (EVs) are envisaged as a promising natural source of nanoparticles to increase outcomes in organ transplantation and control inflammatory diseases. However, poor EV preparations containing highly-abundant soluble proteins may mask genuine vesicular-associated functions and provide misleading data...
2017: Theranostics
https://www.readbyqxmd.com/read/28036315/evaluation-of-the-immunogenic-property-of-nt-h-influenzae-protein-d-with-neisseria-meningitidis-omv-in-balb-c
#7
Ava Behrouzi, Saeid Bouzari, Seyed Davar Siadat, Mana Oloomi, Mehdi Davari, Hoora Mazaheri
INTRODUCTION: Identifying ideal non typeable Haemophilus influenzae (NTHi) vaccine candidates has not been easy due to extensive sequence and antigenic variation among gene products interacting with the immune system. Protein D (PD) is a highly conserved 42 kDa surface lipoprotein available in all H. influenzae, including NTHi. METHODOLOGY: In this study, the gene encoding PD was cloned from H. influenzae and expressed in Escheriachia coli TOPO10 cell in pBAD vector...
December 30, 2016: Journal of Infection in Developing Countries
https://www.readbyqxmd.com/read/28035604/decomplexation-efficiency-and-mechanism-of-cu-ii-edta-by-h2o2-coupled-internal-micro-electrolysis-process
#8
Dongfang Zhou, Yongyou Hu, Qian Guo, Weiguang Yuan, Jiefan Deng, Yapan Dang
Internal micro-electrolysis (IE) coupled with Fenton oxidation (IEF) was a very effective technology for copper (Cu)-ethylenediaminetetraacetic acid (EDTA) wastewater treatment. However, the mechanisms of Cu(2+) removal and EDTA degradation were scarce and lack persuasion in the IEF process. In this paper, the decomplexation and removal efficiency of Cu-EDTA and the corresponding mechanisms during the IEF process were investigated by batch test. An empirical equation and the oxidation reduction potential (ORP) index were proposed to flexibly control IE and the Fenton process, respectively...
December 29, 2016: Environmental Science and Pollution Research International
https://www.readbyqxmd.com/read/28005930/nanoparticle-tracking-analysis-for-the-enumeration-and-characterization-of-mineralo-organic-nanoparticles-in-feline-urine
#9
M Mellema, M Stoller, Y Queau, S P Ho, T Chi, J A Larsen, N Passlack, A J Fascetti, C Mohr, J L Westropp
Urinary stone disease, particularly calcium oxalate, is common in both humans and cats. Calcifying nanoparticles (CNP) are spherical nanocrystallite material, and are composed of proteins (fetuin, albumin) and inorganic minerals. CNP are suggested to play a role in a wide array of pathologic mineralization syndromes including urolithiasis. We documented the development of a clinically relevant protocol to assess urinary CNP in 9 healthy cats consuming the same diet in a controlled environment using Nanoparticle Tracking Analysis (NTA®)...
2016: PloS One
https://www.readbyqxmd.com/read/28003434/gene-cloning-recombinant-expression-purification-and-characterization-of-l-methionine-decarboxylase-from-streptomyces-sp-590
#10
Masaya Hayashi, Akane Okada, Kumiko Yamamoto, Tomomi Okugochi, Chika Kusaka, Daizou Kudou, Michiko Nemoto, Junko Inagaki, Yuu Hirose, Toshihide Okajima, Takashi Tamura, Kenji Soda, Kenji Inagaki
l-Methionine decarboxylase (MetDC) from Streptomyces sp. 590 depends on pyridoxal 5'-phosphate and catalyzes the non-oxidative decarboxylation of l-methionine to produce 3-methylthiopropylamine and carbon dioxide. MetDC gene (mdc) was determined to consist of 1,674 bp encoding 557 amino acids, and the amino acid sequence is similar to that of l-histidine decarboxylases and l-valine decarboxylases from Streptomyces sp. strains. The mdc gene was cloned and recombinant MetDC was heterologously expressed by Escherichia coli The purification of recombinant MetDC was carried out by DEAE-Toyopearl and Ni-NTA agarose column chromatography...
December 21, 2016: Journal of Biochemistry
https://www.readbyqxmd.com/read/27936564/application-of-the-mof-eu-btc-as-a-luminescent-marker-for-gsr-a-synthesis-characterization-and-toxicity-study
#11
Marcella Auxiliadora de Melo Lucena, Marina F L Oliveira, Aline M Arouca, Márcio Talhavini, Eduardo A Ferreira, Severino Alves Júnior, Fabiane H Veiga-Souza, Ingrid Tavora Weber
The 3D metal-organic framework (MOF) [Eu(BTC)] (where BTC = trimesic acid) was synthetized in 20 minutes by a microwave-assisted hydrothermal method with a yield of 89%. A structural and spectroscopic study, performed by X-ray diffraction, thermogravimetry and photoluminescence spectroscopy, showed that this framework has high crystallinity, thermal stability and luminescence. This MOF had a red-orange luminescence when excited with UV radiation (λ = 254 nm) and a high potential for use as a luminescent marker for gunshot residues (GSR)...
December 12, 2016: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/27928487/immunoreactivity-evaluation-of-a-new-recombinant-chimeric-protein-against-brucella-in-the-murine-model
#12
Abbas Abdollahi, Shahla Mansouri, Jafar Amani, Mahdi Fasihi-Ramandi, Mohammad Moradi
BACKGROUND AND OBJECTIVES: Brucellosis is an important health problem in developing countries and no vaccine is available for the prevention of infection in humans. Because of clinically infectious diseases and their economic consequences in human and animals, designing a proper vaccine against Brucella is desirable. In this study, we evaluated the immune responses induced by a designed recombinant chimera protein in murine model. MATERIALS AND METHODS: Three immunodominant antigens of Brucella have been characterized as potential immunogenic and protective antigens including: trigger factor (TF), Omp31 and Bp26 were fused together by EAAAK linkers to produce a chimera (structure were designed in silico), which was synthesized, cloned, and expressed in E...
June 2016: Iranian Journal of Microbiology
https://www.readbyqxmd.com/read/27920885/cloning-and-expression-of-soluble-recombinant-hiv-1-crf35-protease-hp-thioredoxin-fusion-protein
#13
Asaad Azarnezhad, Zohreh Sharifi, Rahmatollah Seyedabadi, Arshad Hosseini, Behrooz Johari, Mahsa Sobhani Fard
BACKGROUND: As a drug target and an antigenic agent, HIV-1 protease (HIV-1 PR) is at the center of attention for designing anti-AIDS inhibitors and diagnostic tests. In previous studies, the production of the recombinant protease has been faced with several difficulties; therefore, the aims of this study were the easy production, purification of the soluble form of protease in E. coli and investigation of its immunoreactivity. METHODS: Protease coding region was isolated from the serum of an infected individual, amplified by RT-PCR and cloned into PTZ57R using TA-cloning...
October 2016: Avicenna Journal of Medical Biotechnology
https://www.readbyqxmd.com/read/27917276/preparation-and-characterization-of-a-novel-nanobody-against-t-cell-immunoglobulin-and-mucin-3-tim-3
#14
Vida Homayouni, Mazdak Ganjalikhani-Hakemi, Abbas Rezaei, Hossein Khanahmad, Mahdi Behdani, Fatemeh Kazemi Lomedasht
OBJECTIVES: As T-cell immunoglobulin and mucin domain 3 (TIM-3) is an immune regulatory molecule; its blocking or stimulating could alter the pattern of immune response towards a desired condition. Based on the unique features of nanobodies, we aimed to construct an anti-TIM-3 nanobody as an appropriate tool for manipulating immune responses for future therapeutic purposes. MATERIALS AND METHODS: We immunized a camel with TIM-3 antigen and then, synthesized a VHH phage: mid library from its B cell's transcriptome using nested PCR...
November 2016: Iranian Journal of Basic Medical Sciences
https://www.readbyqxmd.com/read/27915002/label-free-histamine-detection-with-nanofluidic-diodes-through-metal-ion-displacement-mechanism
#15
Mubarak Ali, Patricio Ramirez, Ivana Duznovic, Saima Nasir, Salvador Mafe, Wolfgang Ensinger
We design and characterize a nanofluidic device for the label-free specific detection of histamine neurotransmitter based on a metal ion displacement mechanism. The sensor consists of an asymmetric polymer nanopore fabricated via ion track-etching technique. The nanopore sensor surface having metal-nitrilotriacetic (NTA-Ni(2+)) chelates is obtained by covalent coupling of native carboxylic acid groups with Nα,Nα-bis(carboxymethyl)-l-lysine (BCML), followed by exposure to Ni(2+) ion solution. The BCML immobilization and subsequent Ni(2+) ion complexation with NTA moieties change the surface charge concentration, which has a significant impact on the current-voltage (I-V) curve after chemical modification of the nanopore...
November 28, 2016: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/27890597/identification-eukaryotic-expression-and-structure-function-characterizations-of-%C3%AE-defensin-like-homologues-from-pelodiscus-sinensis
#16
Haining Yu, Hui Wang, Xuelian Liu, Lan Feng, Xue Qiao, Shasha Cai, Nannan Shi, Yipeng Wang
Defensins are a group of host defense peptides that play a central role in host innate immune responses. Here, 26 genes encoding β-defensin-like peptides have been identified for the first time in Pelodiscus sinensis using database mining approach. Phylogenetic study confirmed that β-defensins are fast evolving genes with high rates of sequence substitutions. The expression level of several selected genes in different tissues was examined by RT-PCR. Ps-BDs mainly adopt β-strands and/or α-helix conformations homology modeled by Rosetta program...
March 2017: Developmental and Comparative Immunology
https://www.readbyqxmd.com/read/27881682/stabilization-of-luciferase-from-renilla-reniformis-using-random-mutations
#17
Megumi Shigehisa, Norie Amaba, Shigeki Arai, Chisato Higashi, Ryo Kawanabe, Ayano Matsunaga, Fina Amreta Laksmi, Masao Tokunaga, Matsujiro Ishibashi
We expressed luciferase (RLuc) from Renilla reniformis in Escherichia coli RLuc was purified using a Ni-NTA column and subsequently characterized. It was unstable in acidic solutions and at 30°C. To increase the stability of RLuc, the Rluc gene was randomly mutated using error-prone polymerase chain reaction. E. coli harboring the mutated gene was screened by detecting luminescence on a plate containing the substrate coelenterazine at 34°C. Three mutants, i.e. N264SS287P, N178D and F116LI137V, were obtained...
January 2017: Protein Engineering, Design & Selection: PEDS
https://www.readbyqxmd.com/read/27881345/-extraction-and-identification-of-semen-derived-exosomes-using-peg6000
#18
Cheng Yang, Wen-Bin Guo, Wan-Song Zhang, Jun Bian, Jian-Kun Yang, Tao Qi, Chun-Yan Wang, Cun-Dong Liu
OBJECTIVE: To extract and identify semen-derived exosome using PEG6000. METHODS: Exosomes were extracted from semen specimens from 6 healthy volunteers with step-by-step centrifugations and ultracentrifugation prior to 8% PEG6000 enrichment. The extracted exosomes were characterized by transmission electron microscopy (TEM), nanoparticle tracking analysis (NTA) and Western blotting. RESULTS: The pellets obtained were round or elliptic membrane vesicles 30 to 150 nm in diameter with intact double membranes and contained low electron density material...
November 20, 2016: Nan Fang Yi Ke da Xue Xue Bao, Journal of Southern Medical University
https://www.readbyqxmd.com/read/27871884/structural-characterization-of-recombinant-human-fibroblast-growth-factor-receptor-2b-kinase-domain-upon-interaction-with-omega-fatty-acids
#19
Masoumeh Moghadasi, Dariush Ilghari, Majid Sirati-Sabet, Abbas Amini, Hamideh Asghari, Nematollah Gheibi
The mutated recombinant kinase domain of human fibroblast growth factor receptor 2b (hFGFR2b) is overexpressed and purified, and its structural changes upon the interaction with three unsaturated fatty acids (UFAs), oleic, linoleic and α-linolenic are studied. This interaction is investigated to find out about the folding and unfolding effect of unsaturated fatty acids on the kinase domain structure of hFGFR2b. Recombinant pLEICS-01 vectors, containing the mutated coding region of hFGFR2b, are expressed in the standard Escherichia coli BL21 (DE3) host cells and purified by Ni(2+)-NTA affinity chromatography...
November 18, 2016: Chemistry and Physics of Lipids
https://www.readbyqxmd.com/read/27864159/high-level-expression-and-efficient-refolding-of-therapeutically-important-recombinant-human-interleukin-3-hil-3-in-e-%C3%A2-coli
#20
Vikas Kumar Dagar, Adivitiya, Yogender Pal Khasa
Human interleukin-3 (hIL-3) is a pleiotropic cytokine that stimulates the differentiation and proliferation of multipotent hematopoietic cells thus making it a therapeutically important molecule. In this study, its poor expression yield was improved by addressing various upstream bottlenecks in E. coli heterologous system. The codon-optimized hIL-3 gene was cloned under various signal sequences and solubility enhancer fusion tags for its hyper-expression under a strong T7 promoter. The optimization of shake flask expression studies resulted in a hIL-3 protein concentration of 225 mg/L in the form of inclusion bodies (IBs)...
November 15, 2016: Protein Expression and Purification
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