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https://www.readbyqxmd.com/read/29739899/novel-xanthomonas-campestris-long-chain-specific-3-oxoacyl-acyl-carrier-protein-reductase-involved-in-diffusible-signal-factor-synthesis
#1
Zhe Hu, Huijuan Dong, Jin-Cheng Ma, Yonghong Yu, Kai-Hui Li, Qiao-Qiao Guo, Chao Zhang, Wen-Bin Zhang, Xinyun Cao, John E Cronan, Haihong Wang
The precursors of the diffusible signal factor (DSF) family signals of Xanthomonas campestris pv. campestris are 3-hydroxyacyl-acyl carrier protein (3-hydroxyacyl-ACP) thioesters having acyl chains of 12 to 13 carbon atoms produced by the fatty acid biosynthetic pathway. We report a novel 3-oxoacyl-ACP reductase encoded by the X. campestris pv. campestris XCC0416 gene ( fabG2 ), which is unable to participate in the initial steps of fatty acyl synthesis. This was shown by the failure of FabG2 expression to allow growth at the nonpermissive temperature of an Escherichia coli fabG temperature-sensitive strain...
May 8, 2018: MBio
https://www.readbyqxmd.com/read/29663661/amino-acid-based-advanced-liquid-formulation-development-for-highly-concentrated-therapeutic-antibodies-balances-physical-and-chemical-stability-and-low-viscosity
#2
Kristina Kemter, Jens Altrichter, Roland Derwand, Thomas Kriehuber, Eva Reinauer, Martin Scholz
To develop highly concentrated therapeutic antibodies enabling convenient subcutaneous application, well stabilizing pharmaceutical formulations with low viscosities are considered to be key. The purpose of this study was to select specific amino acid combinations that reduce and balance aggregation, fragmentation and chemical degradation and also lower viscosity of highly concentrated liquid antibodies. As a model, the therapeutically well-established antibody trastuzumab (25 - >200 mg/mL) in liquid formulation was used...
April 16, 2018: Biotechnology Journal
https://www.readbyqxmd.com/read/29547841/suppressing-autophagy-enhances-disulfiram-copper-induced-apoptosis-in-non-small-cell-lung-cancer
#3
Xi Wu, Xue Xue, Lihui Wang, Wenjing Wang, Jian Han, Xiaoxue Sun, Haotian Zhang, Yueyang Liu, Xiaohang Che, Jingyu Yang, Chunfu Wu
Autophagy, a cellular survival mechanism, is thought to allow the recycling of cellular breakdown products when cancer cells are subjected to chemotherapy, thus decreasing drug-induced apoptosis. Disulfiram (DSF), a drug widely used to control alcoholism, possesses anticancer activity by inducing apoptosis in vitro and in vivo in a copper (Cu)-dependent manner. Our previous studies proved that DSF/Cu exerts increased anti-tumor effects on non-small cell lung cancer (NSCLC) xenograft models, and inhibits NSCLC recurrence driven by ALDH-positive cancer stem cells...
March 13, 2018: European Journal of Pharmacology
https://www.readbyqxmd.com/read/29508082/high-throughput-differential-scanning-fluorimetry-dsf-formulation-screening-with-complementary-dyes-to-assess-protein-unfolding-and-aggregation-in-presence-of-surfactants
#4
Sean M McClure, Patrick L Ahl, Jeffrey T Blue
PURPOSE: The purpose was to evaluate DSF for high throughput screening of protein thermal stability (unfolding/ aggregation) across a wide range of formulations. Particular focus was exploring PROTEOSTAT® - a commercially available fluorescent rotor dye - for detection of aggregation in surfactant containing formulations. Commonly used hydrophobic dyes (e.g. SYPRO™ Orange) interact with surfactants, complicating DSF measurements. METHODS: CRM197 formulations were prepared and analyzed in standard 96-well plate rT-PCR system, using SYPRO™ Orange and PROTEOSTAT® dyes...
March 5, 2018: Pharmaceutical Research
https://www.readbyqxmd.com/read/29428410/fragment-growing-guided-design-of-keap1-nrf2-protein-protein-interaction-inhibitors-for-targeting-myocarditis
#5
Ning Meng, Hua Tang, Hao Zhang, Chengshi Jiang, Li Su, Xiao Min, Wannian Zhang, Hua Zhang, Zhenyuan Miao, Wen Zhang, Chunlin Zhuang
Small-molecule inhibitors that block the Keap1-Nrf2 protein-protein interactions are being intensely pursued as a new therapeutic strategy for oxidative stress-related diseases, such as cancer, diabetes, Alzheimer's disease, arteriosclerosis, inflammation and myocarditis. However, there are not enough studies on antioxidant treatments using small molecules in myocarditis. We herein provided a series of novel hydronaphthoquinones as the Keap1-Nrf2 interaction inhibitors targeting LPS-induced myocarditis both in vitro and in vivo...
March 2018: Free Radical Biology & Medicine
https://www.readbyqxmd.com/read/29423059/brain-and-brain-tumor-penetrating-disulfiram-nanoparticles-sequence-of-cytotoxic-events-and-efficacy-in-human-glioma-cell-lines-and-intracranial-xenografts
#6
Hanumantha Rao Madala, Surendra R Punganuru, Francis Ali-Osman, Ruiwen Zhang, Kalkunte S Srivenugopal
There is great interest in repurposing disulfiram (DSF), a rapidly metabolizing nontoxic drug, for brain cancers and other cancers. To overcome the instability and low therapeutic efficacy, we engineered passively-targeted DSF-nanoparticles (DSFNPs) using biodegradable monomethoxy (polyethylene glycol) d,l-lactic-co-glycolic acid (mPEG-PLGA) matrix. The physicochemical properties, cellular uptake and the blood brain-barrier permeability of DSFNPs were investigated. The DSFNPs were highly stable with a size of ∼70 nm with a >90% entrapment...
January 9, 2018: Oncotarget
https://www.readbyqxmd.com/read/29376540/selective-protein-unfolding-a-universal-mechanism-of-action-for-the-development-of-irreversible-inhibitors
#7
Samuel Askin, Thomas E H Bond, Alanna E Sorenson, Morgane J J Moreau, Helma Antony, Rohan A Davis, Patrick M Schaeffer
High-throughput differential scanning fluorimetry of GFP-tagged proteins (HT-DSF-GTP) was applied for the identification of novel enzyme inhibitors acting by a mechanism termed: selective protein unfolding (SPU). Four different protein targets were interrogated with the same library to identify target-selective hits. Several hits selectively destabilized bacterial biotin protein ligase. Structure-activity relationship data confirmed a structure-dependent mechanism of protein unfolding. Simvastatin and altenusin were confirmed to irreversibly inactivate biotin protein ligase...
February 13, 2018: Chemical Communications: Chem Comm
https://www.readbyqxmd.com/read/29329817/isothermal-chemical-denaturation-as-a-complementary-tool-to-overcome-limitations-of-thermal-differential-scanning-fluorimetry-in-predicting-physical-stability-of-protein-formulations
#8
Hristo Svilenov, Uroš Markoja, Gerhard Winter
Various stability indicating techniques find application in the early stage development of novel therapeutic protein candidates. Some of these techniques are used to select formulation conditions that provide high protein physical stability. Such approach is highly dependent on the reliability of the stability indicating technique used. In this work, we present a formulation case study in which we evaluate the ability of differential scanning fluorimetry (DSF) and isothermal chemical denaturation (ICD) to predict the physical stability of a model monoclonal antibody during accelerated stability studies...
April 2018: European Journal of Pharmaceutics and Biopharmaceutics
https://www.readbyqxmd.com/read/29322563/microfluidics-based-lc-ms-mrm-approach-for-the-relative-quantification-of-burkholderia-cenocepacia-secreted-virulence-factors
#9
Sofie Depluverez, Simon Daled, Stijn De Waele, Sören Planckaert, Jolien Schoovaerts, Dieter Deforce, Bart Devreese
Burkholderia cenocepacia is an opportunistic pathogen that is commonly isolated from patients with cystic fibrosis (CF). Several virulence factors have been identified, including extracellular enzymes that are secreted by type II and type VI secretion systems. The activity of these secretion systems is modulated by quorum sensing. Apart from the classical acylhomoserine lactone quorum sensing, B. cenocepacia also uses the diffusible signal factor system (DSF) i.e. 2-undecenoic acid derivatives that are recognized by specific receptors resulting in changes in biofilm formation, motility and virulence...
January 11, 2018: Rapid Communications in Mass Spectrometry: RCM
https://www.readbyqxmd.com/read/29274360/disulfiram-with-or-without-metformin-inhibits-oesophageal-squamous-cell-carcinoma-in-vivo
#10
Rupal Jivan, Jade Peres, Leonard Howard Damelin, Reubina Wadee, Robin Bruce Veale, Sharon Prince, Demetra Mavri-Damelin
Oesophageal squamous cell carcinoma (OSCC) is highly prevalent in developing countries but there has been little recent progress into efficacious yet affordable treatment strategies. Drug repurposing is one attractive approach for cancer therapy. Disulfiram (DSF), used to treat alcoholism, inhibits cancer growth and we previously found that DSF perturbs protein degradation/turnover pathways in vitro. This was enhanced by combining DSF with the anti-diabetic drug metformin (Met). Here, we investigated DSF with/without Met, against OSCC in vivo...
March 28, 2018: Cancer Letters
https://www.readbyqxmd.com/read/29235198/substrate-interaction-defects-in-histidyl-trna-synthetase-linked-to-dominant-axonal-peripheral-neuropathy
#11
Jamie A Abbott, Rebecca Meyer-Schuman, Vincenzo Lupo, Shawna Feely, Inès Mademan, Stephanie N Oprescu, Laurie B Griffin, M Antonia Alberti, Carlos Casasnovas, Sharon Aharoni, Lina Basel-Vanagaite, Stephan Züchner, Peter De Jonghe, Jonathan Baets, Michael E Shy, Carmen Espinós, Borries Demeler, Anthony Antonellis, Christopher Francklyn
Histidyl-tRNA synthetase (HARS) ligates histidine to cognate tRNA molecules, which is required for protein translation. Mutations in HARS cause the dominant axonal peripheral neuropathy Charcot-Marie-Tooth disease type 2W (CMT2W); however, the precise molecular mechanism remains undefined. Here, we investigated three HARS missense mutations associated with CMT2W (p.Tyr330Cys, p.Ser356Asn, and p.Val155Gly). The three mutations localize to the HARS catalytic domain and failed to complement deletion of the yeast ortholog (HTS1)...
March 2018: Human Mutation
https://www.readbyqxmd.com/read/29194015/phytase-from-citrobacter-koseri-pm-7-enhanced-production-using-statistical-method-and-application-in-ameliorating-mineral-bioaccessibility-and-protein-digestibility-of-high-phytate-food
#12
Preeti Tripathi, Jyothi Lakshmi A, Mukesh Kapoor
The present study was aimed at enhancing phytase (Phy-Ck) production from Citrobacter koseri PM-7 using response surface methodology (RSM) and improving the bioaccessibility of minerals (Fe and Zn) and protein digestibility in high-phytate food using Phy-Ck. A five-variable and three-level central composite design of RSM using wheat bran (6.681%, w/v), inoculum level (2.5%, v/v), and triton X-100 (0.2%, v/v) resulted in up to 5.57-fold (1.047 U/ml) improvement in Phy-Ck yield from C. koseri PM-7 when compared with fermentation media I and II...
January 2, 2018: Preparative Biochemistry & Biotechnology
https://www.readbyqxmd.com/read/29065820/recent-advances-in-antabuse-disulfiram-the-importance-of-its-metal-binding-ability-to-its-anticancer-activity
#13
REVIEW
Maricela Viola-Rhenals, Kush R Patel, Laura Jaimes-Santamaria, Guojun Wu, Jinbao Liu, Q Ping Dou
BACKGROUND: Considerable evidence demonstrates the importance of dithiocarbamates especially disulfiram as anticancer drugs. However there are no systematic reviews outlining how their metal-binding ability is related to their anticancer activity. This review aims to summarize chemical features and metal-binding activity of disulfiram and its metabolite DEDTC, and discuss different mechanisms of action of disulfiram and their contributions to the drug's anticancer activity. METHODS: We undertook a disulfiram-related search on bibliographic databases of peerreviewed research literature, including many historic papers and in vitro, in vivo, preclinical and clinical studies...
February 12, 2018: Current Medicinal Chemistry
https://www.readbyqxmd.com/read/29050650/glycaemic-response-after-intake-of-a-high-energy-high-protein-diabetes-specific-formula-in-older-malnourished-or-at-risk-of-malnutrition-type-2-diabetes-patients
#14
Hamid Laksir, Mirian Lansink, Sophie C Regueme, Johan de Vogel-van den Bosch, Andreas F H Pfeiffer, Isabelle Bourdel-Marchasson
BACKGROUND & AIMS: Several studies with diabetes-specific formulas (DSFs) for hyperglycaemic patients in need of nutritional support have been conducted in non-malnourished patients, mainly comparing products with varying macronutrient compositions. Here, the effect of a high energy, high protein DSF on postprandial responses was compared to a product with a similar macronutrient composition in malnourished or at risk of malnutrition patients with type 2 diabetes. METHODS: In this randomised, double-blind cross-over study, 20 patients were included...
October 6, 2017: Clinical Nutrition: Official Journal of the European Society of Parenteral and Enteral Nutrition
https://www.readbyqxmd.com/read/28986384/mass-spectrometry-for-fragment-screening
#15
REVIEW
Daniel Shiu-Hin Chan, Andrew J Whitehouse, Anthony G Coyne, Chris Abell
Fragment-based approaches in chemical biology and drug discovery have been widely adopted worldwide in both academia and industry. Fragment hits tend to interact weakly with their targets, necessitating the use of sensitive biophysical techniques to detect their binding. Common fragment screening techniques include differential scanning fluorimetry (DSF) and ligand-observed NMR. Validation and characterization of hits is usually performed using a combination of protein-observed NMR, isothermal titration calorimetry (ITC) and X-ray crystallography...
October 6, 2017: Essays in Biochemistry
https://www.readbyqxmd.com/read/28946828/retention-of-the-structure-and-function-of-heparan-sulfate-biomaterials-after-gamma-irradiation
#16
Raymond A A Smith, R J E Chua, Susan M Carnachan, Clarissa L L Tan, Ian M Sims, Simon F R Hinkley, Victor Nurcombe, Simon M Cool
Heparan sulfate (HS) is a highly heterogeneous polysaccharide implicated in many important biological processes. Our previous work has demonstrated that a particular affinity-selected HS (referred to henceforth as "HS3") is capable of enhancing the osteogenic effects of bone morphogenetic protein 2 (BMP2). Here, we gamma-irradiated HS with 26 kGy of ionizing radiation to determine how this affected the structure, composition, and function. Initial structural studies were performed on a commercial preparation of HS as a proof-of-concept...
November 30, 2017: Tissue Engineering. Part A
https://www.readbyqxmd.com/read/28791001/novel-in-vitro-screening-system-based-on-differential-scanning-fluorimetry-to-search-for-small-molecules-against-the-disassembly-or-assembly-of-hiv-1-capsid-protein
#17
Yasuyuki Miyazaki, Naoya Doi, Takaaki Koma, Akio Adachi, Masako Nomaguchi
Varieties of in vitro systems have been used to study biochemical properties of human immunodeficiency virus Gag-capsid protein (HIV Gag-CA). Recently, we have comparatively characterized HIV-1 and HIV-2 Gag-CA proteins using such technology, and have demonstrated that the NaCl-initiated CA-polymerization in vitro and the stability of CA N-terminal domain as judged by differential scanning fluorimetry (DSF) are inversely correlated. In this study, we found that ZnCl2 works as a competent initiator of the in vitro HIV-1 CA-polymerization at much lower concentrations than those of NaCl frequently used for the polymerization initiation...
2017: Frontiers in Microbiology
https://www.readbyqxmd.com/read/28740306/comparative-study-of-the-quality-characteristics-of-defatted-soy-flour-treated-by-supercritical-carbon-dioxide-and-organic-solvent
#18
Sung-Won Kang, M Shafiur Rahman, Ah-Na Kim, Kyo-Yeon Lee, Chan-Yang Park, William L Kerr, Sung-Gil Choi
Defatted soy flour is a potential source of food protein, amino acids, ash and isoflavones. The supercritical carbon dioxide (SC-CO2) and a traditional organic solvent extraction methods were used to remove fat from soy flour, and the quality characteristics of a control soy flour (CSF), defatted soy flour by SC-CO2 (DSFSC-CO2) and defatted soy flour by an organic solvent (DSF-OS) were compared. The SC-CO2 process was carried out at a constant temperature of 45 °C, and a pressure of 40 MPa for 3 h with a CO2 flow rate of 30 g/min...
July 2017: Journal of Food Science and Technology
https://www.readbyqxmd.com/read/28637248/expression-and-comparison-of-codon-optimised-aspergillus-tubingensis-amylase-variants-in-saccharomyces-cerevisiae
#19
Rosemary A Cripwell, Shaunita H Rose, Willem H van Zyl
The expression of codon optimised genes is a popular genetic engineering approach for the production of industrially relevant proteins. This study investigates and compares the expression of codon optimised and codon adapted amylase variants. The Aspergillus tubingensis raw starch hydrolysing α-amylase (amyA) and glucoamylase (glaA) encoding genes were redesigned using synonymous codons and expressed in Saccharomyces cerevisiae Y294. Codon optimisation to favour S. cerevisiae codon bias resulted in a decrease in extracellular enzyme activity of 72% (30...
June 1, 2017: FEMS Yeast Research
https://www.readbyqxmd.com/read/28464377/bile-salts-and-alkaline-ph-reciprocally-modulate-the-interaction-between-the-periplasmic-domains-of-vibrio-cholerae-toxr-and-toxs
#20
Charles R Midgett, Salvador Almagro-Moreno, Maria Pellegrini, Ronald K Taylor, Karen Skorupski, F Jon Kull
ToxR is a transmembrane transcription factor that is essential for virulence gene expression and human colonization by Vibrio cholerae. ToxR requires its operon partner ToxS, a periplasmic integral membrane protein, for full activity. These two proteins are thought to interact through their respective periplasmic domains, ToxRp and ToxSp. In addition, ToxR is thought to be responsive to various environmental cues, such as bile salts and alkaline pH, but how these factors influence ToxR is not yet understood...
July 2017: Molecular Microbiology
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