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https://www.readbyqxmd.com/read/28540175/cloning-and-expression-of-three-thaumatin-like-protein-genes-from-polyporus-umbellatus
#1
Mengmeng Liu, Dawei Zhang, Yongmei Xing, Shunxing Guo
Genes encoding thaumatin-like protein (TLPs) are frequently found in fungal genomes. However, information on TLP genes in Polyporus umbellatus is still limited. In this study, three TLP genes were cloned from P. umbellatus. The full-length coding sequence of PuTLP1, PuTLP2 and PuTLP3 were 768, 759 and 561 bp long, respectively, encoding for 256, 253 and 187 amino acids. Phylogenetic trees showed that P. umbellatus PuTLP1, PuTLP2 and PuTLP3 were clustered with sequences from Gloeophyllum trabeum, Trametes versicolor and Stereum hirsutum, respectively...
May 2017: Acta Pharmaceutica Sinica. B
https://www.readbyqxmd.com/read/28540060/microparticle-formation-in-peritoneal-dialysis-a-proof-of-concept-study
#2
Shareef Akbari, Rima Abou-Arkoub, Suzy Sun, Swapnil Hiremath, Arkadiy Reunov, Brendan B McCormick, Marcel Ruzicka, Dylan Burger
BACKGROUND: Injury to the mesothelial layer of the peritoneal membrane during peritoneal dialysis (PD) is implicated in loss of ultrafiltration capacity, but there are no validated biomarkers for mesothelial cell injury. Microparticles (MPs) are 0.1 to 1.0 µm membrane vesicles shed from the cell surface following injury and are sensitive markers of tissue damage. Formation of MPs in the peritoneal cavity during PD has not been reported to date. METHODS: We designed a single-center, proof of concept study to assess whether peritoneal solution exposure induces formation of mesothelial MPs suggestive of PD membrane injury...
2017: Canadian Journal of Kidney Health and Disease
https://www.readbyqxmd.com/read/28538167/understanding-the-bioavailability-and-sequestration-of-different-metal-cations-in-the-presence-of-a-biodegradable-chelant-mgda-in-biological-fluids-and-natural-waters
#3
Clemente Bretti, Rosalia Maria Cigala, Concetta De Stefano, Gabriele Lando, Silvio Sammartano
Thermodynamic information about the metal-ligand interaction between Fe(3+), Zn(2+), Cu(2+) and Sn(2+), and a biodegradable ligand as MGDA is reported. The speciation scheme was obtained by means of potentiometric measurements and isothermal titration calorimetry (to determine enthalpy changes) in NaCl medium. The formation of the ML and MLOH species was evidenced for all the metal cations, and for Fe(3+) also the ML2 and ML(OH)2 were found. The relative stability, for the ML species, follows the order: Sn(2+) > Fe(3+) > Cu(2+) > Zn(2+)...
May 16, 2017: Chemosphere
https://www.readbyqxmd.com/read/28531338/placental-exosomes-as-early-biomarker-of-preeclampsia-potential-role-of-exosomal-micrornas-across-gestation
#4
Carlos Salomon, Dominic Guanzon, Katherin Scholz-Romero, Sherri Longo, Paula Correa, Sebastian E Illanes, Gregory E Rice
Context: There is a need to develop strategies for early prediction of patients who will develop preeclampsia (PE), in order to establish preventive strategies to reduce the prevalence and severity of the disease and their associated complications. Objective: The objective of this study was to investigate whether exosomes and their microRNA cargo present in maternal circulation can be used as early biomarker for PE. Design, Setting, Patients, and Interventions: A retrospective stratified study design was used to quantify total exosomes (EXO) and placenta-derived exosomes (PdE) present in maternal plasma of normal (n=32 per time point) and PE (n=15 per time point) pregnancies...
May 22, 2017: Journal of Clinical Endocrinology and Metabolism
https://www.readbyqxmd.com/read/28526203/quantitative-evaluation-of-inhibitory-effect-of-various-substances-on-anaerobic-ammonia-oxidation-anammox
#5
Tomotaka Nakamura, Yuhki Harigaya, Yuya Kimura, Megumi Kuroiwa, Yuhri Kurata, Kazuichi Isaka, Yuichi Suwa
The inhibitory effect of 20 substances of various chemical species on the anaerobic ammonia oxidation (anammox) activity of an enrichment culture, predominated by Candidatus Brocadia, was determined systematically by using a (15)N tracer technique. The initial anammox rate was determined during first 25 min with a small-scale anaerobic batch incubation supplemented with possible inhibitors. Although Cu(2+) and Mn(2+) did not inhibit anammox, the remaining 18 substances [Ni(2+), Zn(2+), Co(2+), [Formula: see text] , Fe(2+), 4 amines, ethylenediaminetetraacetic acid (EDTA), ethylenediamine-N,N'-bis (2-hydroxyphenylacetic acid) (EDDHA), citric acid, nitrilotriacetic acid (NTA), N,N-dimethylacetamide (DMA), 1,4-dioxane, dimethyl sulfoxide (DMSO), N,N-dimethylformamide (DMF) and tetrahydrofuran (THF)] were inhibitory...
May 16, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28523920/expression-of-bovine-rotavirus-vp8-and-preparation-of-igy-antibodies-against-recombinant-vp8
#6
H S Kim, B Lee, S Y Han, Y T Jung
Group A rotavirus is the leading cause of acute gastroenteritis in cattle and swine. Although, vaccination against this virus is an effective strategy for prevention, additional strategy to control disease is necessary. Egg yolk immunoglobulin (IgY)-based passive immunization could be a better option in preventing this disease. Bovine rotavirus (BRV) is group A rotavirus and possesses a genome of 11 segments of double-stranded RNA. The outer layer of capsid is composed of two proteins (VP7 and VP4), which induce virus neutralizing antibodies...
2017: Acta Virologica
https://www.readbyqxmd.com/read/28522197/identification-of-the-upstream-4-chlorophenol-biodegradation-pathway-using-a-recombinant-monooxygenase-from-arthrobacter-chlorophenolicus-a6
#7
Su Yeon Cho, Oh Sung Kwean, Jun Won Yang, Wooyoun Cho, Seonyeong Kwak, Sungyoon Park, Yejee Lim, Han S Kim
This study aimed to clarify the initial 4-chlorophenol (4-CP) biodegradation pathway promoted by a two-component flavin-diffusible monooxygenase (TC-FDM) consisting of CphC-I and CphB contained in Arthrobacter chlorophenolicus A6 and the decomposition function of CphC-I. The TC-FDM genes were cloned from A. chlorophenolicus A6, and the corresponding enzymes were overexpressed. Since CphB was expressed in an insoluble form, Fre, a flavin reductase obtained from Escherichia coli, was used. These enzymes were purified using Ni(2+)-NTA resin...
May 4, 2017: Bioresource Technology
https://www.readbyqxmd.com/read/28503428/soluble-expression-of-recomb-inant-cmyc-klf4-oct4-and-sox2-proteins-in-bacteria-and-transduction-into-living-cells
#8
Guo-Dan Liu, Shi-Feng Zhou, Xu-Chen Ding, Chun-Lai Fang, Shu-Yong Mi, Xiang-Chun Gao, Qing Han
AIM: To develop a new method to produce recombinant reprogramming proteins, cMyc, Klf4, Oct4, and Sox2, in soluble format with low cost for the generation of induced pluripotent stem cells (iPSCs). METHODS: A short polypeptide sequence derived from the HIV trans-activator of transcription protein (TAT) and the nucleus localization signal (NLS) polypeptide were fused to the N terminus of the reprogramming proteins and they were constructed into pCold-SUMO vector which can extremely improve the solubility of recombinant proteins...
2017: International Journal of Ophthalmology
https://www.readbyqxmd.com/read/28499338/hyaluronic-acid-molecular-weight-dependent-modulation-of-mucin-nanostructure-for-potential-mucosal-therapeutic-applications
#9
Irene M Hansen, Morten F Ebbesen, Liselotte Kaspersen, Troels Thomsen, Konrad Bienk, Yunpeng Cai, Birgitte Mølholm Malle, Kenneth A Howard
This study investigates the effects of different molecular weight hyaluronic acids (HAs) on the mucosal nanostructure using a pig stomach mucin hydrogel as a mucosal barrier model. Microparticles (1.0 μm) and nanoparticles (200 nm) were used as probes, and their movement in mucin was studied by a three-dimensional confocal microscopy-based particle tracking technique and by Nanoparticle Tracking Analysis (NTA) after addition of high-molecular weight (900 kDa) and low-molecular weight (33 kDa) HA. This demonstrated a molecular weight-dependent HA modulation of the mucin nanostructure with a 2...
May 26, 2017: Molecular Pharmaceutics
https://www.readbyqxmd.com/read/28484911/molecular-cloning-and-expression-of-a-glycosyltransferase-from-bacillus-subtilis-for-modification-of-morin-and-related-polyphenols
#10
Qianqian Wang, Yixiang Xu, Jiaqi Xu, Xudong Wang, Chen Shen, Yan Zhang, Xiufeng Liu, Boyang Yu, Jian Zhang
OBJECTIVES: To characterize glycosyltransferases from Bacillus subtilis ATCC 6633 and investigate their substrate specificity towards plant polyphenols. RESULTS: Among the cloned and expressed six UDP-glycosyltransferases (BsGT1-6), BsGT-1 showed activity with a wide range of polyphenols: morin, quercetin, alizarin, rehin, curcumin and aloe emodin. The gene of BsGT-1 has an ORF of 1206 bp encoding 402 amino acids. The recombinant enzyme was purified to homogeneity by Ni-NTA affinity chromatograph, and its biochemical characteristics were identified by HPLC-UV/MS, (1)H-NMR and (13)C-NMR...
May 8, 2017: Biotechnology Letters
https://www.readbyqxmd.com/read/28477005/biosynthesis-of-zinc-oxide-nanoparticles-by-petals-extract-of-rosa-indica-l-its-formulation-as-nail-paint-and-evaluation-of-antifungal-activity-against-fungi-causing-onychomycosis
#11
Nikita Tiwari, Raksha Pandit, Swapnil Gaikwad, Aniket Gade, Mahendra Rai
Aim: The authors report the biological synthesis of zinc oxide nanoparticles (ZnO-NPs) from the petals extract of Rosa indica L. (rose). Its efficacy was evaluated against two dermatophytes: namely: Trichophyton mentagrophytes and Microsporum canis which cause onychomycosis. The activity of antibiotics against the tested dermatophytes was enhanced, when evaluated in combination with ZnO-NPs. Methods and results: The synthesised ZnO-NPs were preliminary detected by using ultraviolet UV visible spectroscopy, which showed specific absorbance...
March 2017: IET Nanobiotechnology
https://www.readbyqxmd.com/read/28476462/new-insights-into-the-antioxidant-activity-and-cytotoxicity-of-arzanol-and-effect-of-methylation-on-its-biological-properties
#12
Antonella Rosa, Angela Atzeri, Mariella Nieddu, Giovanni Appendino
The heterodimeric phloroglucinyl pyrone arzanol (Arz) has raised considerable interest because of its antiviral, anti-inflammatory, and antioxidant activity. We have investigated the effect of methylation of the pyrone moiety on the antioxidant activity and cytotoxicity of Arz. This manoeuvre, that left the polyphenolic moiety unscathed, was nevertheless detrimental for antioxidant activity in both the cholesterol thermal degradation- and the Cu(2+)-induced liposome oxidation assays, providing evidence of structure-activity relationships that go beyond the preservation of the polyphenolic pharmacophore...
May 3, 2017: Chemistry and Physics of Lipids
https://www.readbyqxmd.com/read/28476278/reliability-and-validity-of-the-treatment-outcome-profile-among-patients-attending-methadone-maintenance-treatment-programs-in-kunming-china
#13
Mei Wang, Jiucheng Shen, Xianling Liu, Yuan Deng, Jiahua Li, Emily Finch, Kim Wolff
BACKGROUND & OBJECTIVE: Substance misuse has been a major health and social issue worldwide and has become an important public health issue in China over the past two decades. Methadone maintenance treatment (MMT) has been proved worldwide by large bodies of research to be one of the most effective practices for illicit drug users. The Treatment Outcome Profile (TOP) was developed in 2007 by the UK National Treatment Agency (NTA). It has been proved to be a reliable instrument for outcome measure...
June 2017: Journal of Substance Abuse Treatment
https://www.readbyqxmd.com/read/28473885/prospects-and-limitations-of-antibody-mediated-clearing-of-lipoproteins-from-blood-plasma-prior-to-nanoparticle-tracking-analysis-of-extracellular-vesicles
#14
Morten Mørk, Aase Handberg, Shona Pedersen, Malene M Jørgensen, Rikke Bæk, Morten K Nielsen, Søren R Kristensen
Introduction: Nanoparticle tracking analysis (NTA) enables measurement of extracellular vesicles (EVs) but lacks the ability to distinct between EVs and lipoproteins which are abundantly present in blood plasma. Limitations in ultracentrifugation and size exclusion chromatography applied for EV isolation may result in inadequate EV purification and preservation. In this proof of concept study, we aimed to evaluate the potential of antibody-mediated removal of lipoproteins from plasma prior to extracellular vesicle (EV) analysis by nanoparticle tracking analysis (NTA)...
2017: Journal of Extracellular Vesicles
https://www.readbyqxmd.com/read/28468076/-recombinant-expression-of-hantaan-virus-protein-n-with-application-of-western-blot-in-detecting-anti-hantavirus-antibody
#15
P P Yao, F Xu, Y S Sun, Z R Yang, Y Zhang, M Yue, H P Zhu
Objective: S gene of hantavirus(HV) was expressed in insect cells by genetic engineering technology. The expression product of S gene was used as antigen to detect anti-HV specific antibody IgG in serum. Methods: Gene encoding NP of the strain HV-Z10 was amplified by PCR and then its eukaryotic expression system rBAC-Z10S-TN was constructed by using the routine genetic engineering method. SDS-PAGE was applied to measure the expression of rNP.Ion-exchange plus Ni-NTA-affinity chromatography was performed to purify the recombinant product...
April 10, 2017: Zhonghua Liu Xing Bing Xue za Zhi, Zhonghua Liuxingbingxue Zazhi
https://www.readbyqxmd.com/read/28460618/specific-targeting-of-her2-positive-head-and-neck-squamous-cell-carcinoma-line-hn5-by-idarubicin-zher2-affibody-conjugate
#16
Marzieh Ghanemi, Aminollah Pourshohod, Mohammad Ali Ghaffari, Alireza Kheirollah, Mansour Amin, Majid Zeinali, Mostafa Jamalan
Expression of human epidermal growth factor receptor type 2 (HER2) in head and neck squamous cell carcinoma (HNSCC) cell line HN5 can be employed with great opportunities of success for specific targeting of anti-cancer chemotherapeutic agents. In the current study, HER2-specific affibody molecule, ZHER2:342 (an engineered protein with great affinity for HER2 receptors) was selected for conjugation to idarubicin (an anti-neoplastic antibiotic). ZHER2:342 affibody gene with one added cysteine code at the its 5' end was synthesized de novo and then inserted into pET302 plasmid and transferred to E...
April 26, 2017: Current Cancer Drug Targets
https://www.readbyqxmd.com/read/28456803/expression-and-characterization-of-serotype-2-streptococcus-suis-arginine-deiminase
#17
Krissana Maneerat, Suganya Yongkiettrakul, Surasak Jiemsup, Pongsri Tongtawe, Marcelo Gottschalk, Potjanee Srimanote
BACKGROUND: Arginine deiminase (ArcA) has been speculated to facilitate the intracellular survival of Streptococcus suis under acidic conditions. However, the physical and biological properties and function of SS2-ArcA have not yet been elucidated. METHODS: Recombinant SS2-ArcA (rSS2-ArcA) was expressed and purified using Ni-NTA affinity chromatography. Under various pH and temperature conditions, the enzymatic properties of purified rSS2-ArcA and crude native SS2-ArcA were determined...
April 29, 2017: Journal of Molecular Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28451120/total-chemical-synthesis-of-proteins-without-hplc-purification
#18
S F Loibl, Z Harpaz, R Zitterbart, O Seitz
The total chemical synthesis of proteins is a tedious and time-consuming endeavour. The typical steps involve solid phase synthesis of peptide thioesters and cysteinyl peptides, native chemical ligation (NCL) in solution, desulfurization or removal of ligation auxiliaries in the case of extended NCL as well as many intermediary and final HPLC purification steps. With an aim to facilitate and improve the throughput of protein synthesis we developed the first method for the rapid chemical total on-resin synthesis of proteins that proceeds without a single HPLC-purification step...
November 1, 2016: Chemical Science
https://www.readbyqxmd.com/read/28450245/heterologous-expression-and-biochemical-studies-of-a-thermostable-glucose-tolerant-%C3%AE-glucosidase-from-methylococcus-capsulatus-bath-strain
#19
Sneha S Sathe, Surabhi Soni, Vikas P Ranvir, Vikram G Choudhari, Annamma A Odaneth, Arvind M Lali, Sanjeev K Chandrayan
Glucose inhibition of β-glucosidase (BG) is a bottleneck in biomass hydrolysis. In this study, a glucose resistant GH1 β-glucosidase gene- Mbgl from Methylococcus capsulatus (bath strain) was cloned and overexpressed in E.coli. The Ni-NTA affinity purified Mbgl displayed an optimum temperature of 70°C and optimum pH was 6.0. The calculated KM of the enzyme was 48.6mM and 0.12mM for cellobiose and 4-Nitrophenyl β-d-glucopyranoside (PNPG) respectively. PNPG hydrolysis in presence of various glucose concentrations showed that the enzyme was stimulated by ∼2...
April 24, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28446197/re-directing-bacterial-microcompartment-systems-to-enhance-recombinant-expression-of-lysis-protein-e-from-bacteriophage-%C3%AF-x174-in-escherichia-coli
#20
Mimi C Yung, Feliza A Bourguet, Timothy S Carpenter, Matthew A Coleman
BACKGROUND: Recombinant expression of toxic proteins remains a challenging problem. One potential method to shield toxicity and thus improve expression of these proteins is to encapsulate them within protein compartments to sequester them away from their targets. Many bacteria naturally produce so-called bacterial microcompartments (BMCs) in which enzymes comprising a biosynthetic pathway are encapsulated in a proteinaeous shell, which is in part thought to shield the cells from the toxicity of reaction intermediates...
April 26, 2017: Microbial Cell Factories
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