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https://www.readbyqxmd.com/read/28813685/purification-and-characterization-of-two-novel-peroxidases-from-the-dye-decolorizing-fungus-bjerkandera-adusta-strain-cx-9
#1
Khelifa Bouacem, Hatem Rekik, Nadia Zaraî Jaouadi, Bilal Zenati, Sidali Kourdali, Mohamed El Hattab, Abdelmalek Badis, Rachid Annane, Samir Bejar, Hocine Hacene, Amel Bouanane-Darenfed, Bassem Jaouadi
Two extracellular peroxidases from Bjerkandera adusta strain CX-9, namely a lignin peroxidase (called LiP BA45) and manganese peroxidase (called MnP BA30), were purified simultaneously by applying successively, ammonium sulfate precipitation-dialysis, Mono-S Sepharose anion-exchange and Sephacryl S-200 gel filtration and biochemically characterized. The sequence of their NH2-terminal amino acid residues showed high homology with those of fungi peroxidases. MALDI-TOF/MS analysis revealed that the purified enzymes MnP BA30 and LiP BA45 were a monomers with a molecular masses 30125...
August 13, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28813594/facile-purification-of-milligram-to-gram-quantities-of-condensed-tannins-according-to-mean-degree-of-polymerization-and-flavan-3-ol-subunit-composition
#2
Ron Herbert Brown, Irene Mueller-Harvey, Wayne E Zeller, Laurie Reinhardt, Elisabetta Stringano, An Gea, Chris Drake, Honorata M Ropiak, Christos Fryganas, Aina Ramsay, Emily E Hardcastle
Unambiguous investigation of condensed tannin (CT) structure-activity relationships in biological systems requires well-characterized, high-purity CTs. Sephadex LH-20 and Toyopearl HW-50F resins were compared for separating CTs from acetone/water extracts and column fractions analyzed for flavan-3-ol subunits, mean degree of polymerization (mDP) and purity. Toyopearl HW-50F generated fractions with higher mDP values and better separation of procyanidins (PC) and prodelphinidins (PD) but required a pre-purification step, more time for large scale purifications and gave poorer recoveries...
August 16, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28813479/metabolomic-profiling-of-amines-in-sepsis-predicts-changes-in-nos-canonical-pathways
#3
Abel Tesfai, Niall MacCallum, Nicholas S Kirkby, Hime Gashaw, Nicola Gray, Elizabeth Want, Gregory J Quinlan, Sharon Mumby, James M Leiper, Mark Paul-Clark, Blerina Ahmetaj-Shala, Jane A Mitchell
RATIONALE: Nitric oxide synthase (NOS) is a biomarker/target in sepsis. NOS activity is driven by amino acids, which cycle to regulate the substrate L-arginine in parallel with cycles which regulate the endogenous inhibitors ADMA and L-NMMA. The relationship between amines and the consequence of plasma changes on iNOS activity in early sepsis is not known. OBJECTIVE: Our objective was to apply a metabolomics approach to determine the influence of sepsis on a full array of amines and what consequence these changes may have on predicted iNOS activity...
2017: PloS One
https://www.readbyqxmd.com/read/28812349/purification-and-characterization-of-latent-polyphenol-oxidase-from-apricot-prunus-armeniaca-l
#4
Ala Eddine Derardja, Matthias Pretzler, Ioannis Kampatsikas, Malika Barkat, Annette Rompel
Polyphenol oxidase from apricot (Prunus armeniaca) (PaPPO) was purified in its latent form (L-PaPPO) and the molecular weight was determined to 63 kDa by SDS-PAGE. L-PaPPO was activated in the presence of substrate at low pH. The activity was enhanced by CuSO4 and low concentrations (≤ 2 mM) of SDS. PaPPO has its pH and temperature optimum at pH 4.5 and 45 °C for catechol as the substrate. It showed diphenolase activity and highest affinity toward 4-methylcatechol (KM = 2.0 mM) and chlorogenic acid (KM = 2...
August 16, 2017: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/28812178/sludge-quantification-at-water-treatment-plant-and-its-management-scenario
#5
Tarique Ahmad, Kafeel Ahmad, Mehtab Alam
Large volume of sludge is generated at the water treatment plants during the purification of surface water for potable supplies. Handling and disposal of sludge require careful attention from civic bodies, plant operators, and environmentalists. Quantification of the sludge produced at the treatment plants is important to develop suitable management strategies for its economical and environment friendly disposal. Present study deals with the quantification of sludge using empirical relation between turbidity, suspended solids, and coagulant dosing...
August 15, 2017: Environmental Monitoring and Assessment
https://www.readbyqxmd.com/read/28811661/myst2-kat7-histone-acetyltransferase-interaction-proteomics-reveals-tumour-suppressor-niam-as-a-novel-binding-partner-in-embryonic-stem-cells
#6
Mercedes Pardo, Lu Yu, Shihpei Shen, Peri Tate, Daniel Bode, Blake L Letney, Dawn E Quelle, William Skarnes, Jyoti S Choudhary
MYST histone acetyltransferases have crucial functions in transcription, replication and DNA repair and are hence implicated in development and cancer. Here we characterise Myst2/Kat7/Hbo1 protein interactions in mouse embryonic stem cells by affinity purification coupled to mass spectrometry. This study confirms that in embryonic stem cells Myst2 is part of H3 and H4 histone acetylation complexes similar to those described in somatic cells. We identify a novel Myst2-associated protein, the tumour suppressor protein Niam (Nuclear Interactor of ARF and Mdm2)...
August 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28811266/expression-and-purification-of-a-difficult-sarcomeric-protein-telethonin
#7
Huanbo Tan, Wencheng Su, Pengju Wang, Wenyu Zhang, Michael Sattler, Peijian Zou
Telethonin anchors the N-terminal region of titin in the Z-disk of the sarcomere by binding to two immunoglobulin-like (Ig) domains (Z1 and Z2) of titin (Z1Z2). Thereby telethonin plays an important role in myofibril assembly and in muscle development and functional regulation. The expression and purification of recombinant telethonin is very challenging. In previous studies, recombinant telethonin expressed from E. coli was refolded in the presence of Z1Z2. Here, we report various strategies to establish a reliable and efficient protocol for the preparation of telethonin and titin Z1Z2 protein...
August 12, 2017: Protein Expression and Purification
https://www.readbyqxmd.com/read/28811204/heterologous-expression-and-biochemical-characterization-of-a-novel-thermostable-sclerotinia-sclerotiorum-gh45-endoglucanase-in-pichia-pastoris
#8
Haifa Chahed, Mohamed Boumaiza, Aymen Ezzine, M Nejib Marzouki
Enzymatic saccharification of lignocellulosic biomass has been widely studied. Mainly endoglucanases were found to be a prerequisite for the quick initial biomass liquefaction. In the present study, Pichia pastoris was used as a host for the heterologous expression of a Sclerotinia sclerotiorum GH45 endoglucanase, Endo2. The recombinant plasmid pPICZαA was used to transform Pichia pastoris. Pichia culture supernatants expressing the recombinant Endo2 (rEndo2) were used for the purification and biochemical characterization of this enzyme...
August 12, 2017: International Journal of Biological Macromolecules
https://www.readbyqxmd.com/read/28811070/evaluation-of-sequence-variability-in-hiv-1-gp41-c-peptide-helix-grafted-proteins
#9
Rachel L Tennyson, Susanne N Walker, Terumasa Ikeda, Reuben S Harris, Brian R McNaughton
Many therapeutically-relevant protein-protein interactions (PPIs) have been reported that feature a helix and helix-binding cleft at the interface. Given this, different approaches to disrupting such PPIs have been developed. While short peptides (<15 amino acids) typically do not fold into a stable helix, researchers have reported chemical approaches to constraining helix structure. However, these approaches rely on laborious, and often expensive, chemical synthesis and purification. Our premise is that protein-based solutions that stabilize a therapeutically-relevant helix offer a number of advantages...
August 1, 2017: Bioorganic & Medicinal Chemistry
https://www.readbyqxmd.com/read/28810509/spatiotemporal-assessment-and-trade-offs-of-multiple-ecosystem-services-based-on-land-use-changes-in-zengcheng-china
#10
Xiao Sun, Feng Li
Driven by rapid urbanization, land use change has become a significant factor influencing ecosystem services (ESs). To support the decision-making process of city planners and policy makers, assessing the spatiotemporal changes associated with multiple ESs is vital. In this study, we developed a general structure to assess the changes of multiple ESs in Zengcheng, China. A new index also was developed to measure the comprehensive ecosystem service (CES). Trade-offs of various ESs were analyzed by using correlation analysis...
August 8, 2017: Science of the Total Environment
https://www.readbyqxmd.com/read/28809985/a-magnetic-micropore-chip-for-rapid-1-hour-unbiased-circulating-tumor-cell-isolation-and-in-situ-rna-analysis
#11
Jina Ko, Neha Bhagwat, Stephanie S Yee, Taylor Black, Colleen Redlinger, Janae Romeo, Mark O'Hara, Arjun Raj, Erica L Carpenter, Ben Z Stanger, David Issadore
The use of microtechnology for the highly selective isolation and sensitive detection of circulating tumor cells has shown enormous promise. One challenge for this technology is that the small feature sizes - which are the key to this technology's performance - can result in low sample throughput and susceptibility to clogging. Additionally, conventional molecular analysis of CTCs often requires cells to be taken off-chip for sample preparation and purification before analysis, leading to the loss of rare cells...
August 15, 2017: Lab on a Chip
https://www.readbyqxmd.com/read/28809830/overexpression-and-purification-of-human-cis-prenyltransferase-in-escherichia-coli
#12
Ilan Edri, Michal Goldenberg, Michal Lisnyansky, Roi Strulovich, Hadas Newman, Anat Loewenstein, Daniel Khananshvili, Moshe Giladi, Yoni Haitin
Prenyltransferases (PT) are a group of enzymes that catalyze chain elongation of allylic diphosphate using isopentenyl diphosphate (IPP) via multiple condensation reactions. DHDDS (dehydrodolichyl diphosphate synthase) is a eukaryotic long-chain cis-PT (forming cis double bonds from the condensation reaction) that catalyzes chain elongation of farnesyl diphosphate (FPP, an allylic diphosphate) via multiple condensations with isopentenyl diphosphate (IPP). DHDDS is of biomedical importance, as a non-conservative mutation (K42E) in the enzyme results in retinitis pigmentosa, ultimately leading to blindness...
August 3, 2017: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/28808974/isolation-purification-and-culture-of-primary-murine-sensory-neurons
#13
Sarah Katzenell, Jorge R Cabrera, Brian J North, David A Leib
Cultured primary neurons have been of extraordinary value for the study of neuronal anatomy, cell biology, and physiology. While use of neuronal cell lines has ease and utility, there are often caveats that arise due to their mitotic nature. This methods article presents detailed methodology for the preparation, purification, and culture of adult murine sensory neurons for the study of herpes simplex virus lytic and latent infections. While virology is the application for our laboratory, these cultures also have broad utility for neurobiologists and cell biologists...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28808967/purification-of-cyclic-gmp-amp-from-viruses-and-measurement-of-its-activity-in-cell-culture
#14
Alice Mayer, Jonathan Maelfait, Anne Bridgeman, Jan Rehwinkel
Sensing of cytoplasmic DNA by cGAS is essential for the initiation of immune responses against several viruses. cGAS also plays important roles in some autoinflammatory and autoimmune diseases and may be involved in immune responses targeting cancer cells. Once activated, cGAS catalyzes the formation of the di-nucleotide 2'-3'-cyclic GMP-AMP (cGAMP), which propagates a signaling cascade leading to the production of type I interferons (IFNs). Interestingly, cGAMP is incorporated into enveloped viruses and is transferred to newly infected cells by virions...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28808944/effect-of-ha330-resin-directed-hemoadsorption-on-a-porcine-acute-respiratory-distress-syndrome-model
#15
Xuefeng Xu, Chune Jia, Sa Luo, Yanming Li, Fei Xiao, Huaping Dai, Chen Wang
BACKGROUND: Blood purification is an emerging approach to dampening the cytokine storm. This study aims to assess the efficacy of HA330 resin-directed hemoadsorption (HA) on endotoxin-induced porcine acute respiratory distress syndrome (ARDS) model. METHODS: Twenty-four Chinese domestic pigs were allocated into saline group receiving intravenous infusion of saline (N = 6) and endotoxin group receiving intravenous infusion of LPS (N = 18). When ALI model was initially diagnosed, six pigs in the LPS and saline group were killed for BALF and histopathological analysis...
August 14, 2017: Annals of Intensive Care
https://www.readbyqxmd.com/read/28808330/generation-and-characterization-of-a-mouse-line-for-monitoring-translation-in-dopaminergic-neurons
#16
Joseph D Dougherty
We developed a mouse line targeting midbrain dopamine neurons for Translating Ribosome Affinity Purification(TRAP). Here, we briefly report on the basic characterization of this mouse line including confirmation of expression of the transgene in midbrain dopamine neurons and validation of its effectiveness in capturing mRNA from these cells. We also report a translational profile of these neurons which may be of use to investigators studying the gene expression of these cells. Finally, we have provided the line to Jackson Laboratories for distribution and use in future studies...
August 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28808309/defluoridation-technology-for-drinking-water-and-tea-by-green-synthesized-fe3o4-al2o3-nanoparticles-coated-polyurethane-foams-for-rural-communities
#17
Sonu Kumari, Suphiya Khan
Fluoride (F) contaminated ground water poses a serious public health concern to rural population with unaffordable purification technologies. Therefore, development of a cost-effective, portable, environment and user-friendly defluoridation technique is imperative. In the present study, we report on the development of a green and cost-effective method that utilizes Fe3O4 and Al2O3 nanoparticles (NPs) that were synthesized using jojoba defatted meal. These NPs were impregnated on to polyurethane foam (PUF) and made into tea infusion bags...
August 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28808144/particulate-matter-exposure-and-stress-hormone-levels-a-randomized-double-blind-crossover-trial-of-air-purification
#18
Huichu Li, Jing Cai, Renjie Chen, Zhuohui Zhao, Zhekang Ying, Lin Wang, Jianmin Chen, Ke Hao, Patrick L Kinney, Honglei Chen, Haidong Kan
BACKGROUND: Exposure to ambient particulate matter (PM) is associated with a number of adverse health outcomes, but potential mechanisms are largely unknown. Metabolomics represents a powerful approach to study global metabolic changes in response to environmental exposures. We therefore conducted this study to investigate changes in serum metabolites in response to the reduction of PM exposure among healthy college students. METHODS: We conducted a randomized, double-blind crossover trial in 55 healthy college students in Shanghai, China...
August 15, 2017: Circulation
https://www.readbyqxmd.com/read/28807561/mass-spectrometry-analysis-of-intact-francisella-bacteria-identifies-lipid-a-structure-remodeling-in-response-to-acidic-ph-stress
#19
Camille B Robert, Michael Thomson, Alain Vercellone, Francesca Gardner, Robert K Ernst, Gérald Larrouy-Maumus, Jérôme Nigou
Structural modification of lipid A, the lipid anchor of LPS, is one of the strategies used by Gram-negative bacteria to evade host innate immunity. Francisella tularensis is a human pathogen that infects and replicates within phagocytic cells. It produces an atypical lipid A, whose structure precludes an efficient recognition by both innate immune players, TLR4 and cationic antimicrobial peptides. Interestingly, a recent report indicates that the lipid A of Francisella (LVS vaccinal strain) undergoes polar modifications when bacteria are grown in human macrophages as compared to in broth...
August 11, 2017: Biochimie
https://www.readbyqxmd.com/read/28807549/a-multiplexed-microfluidic-toolbox-for-the-rapid-optimization-of-affinity-driven-partition-in-aqueous-two-phase-systems
#20
Eduardo J S Bras, Ruben R G Soares, Ana M Azevedo, Pedro Fernandes, Miguel Arévalo-Rodríguez, Virginia Chu, João P Conde, M Raquel Aires-Barros
Antibodies and other protein products such as interferons and cytokines are biopharmaceuticals of critical importance which, in order to be safely administered, have to be thoroughly purified in a cost effective and efficient manner. The use of aqueous two-phase extraction (ATPE) is a viable option for this purification, but these systems are difficult to model and optimization procedures require lengthy and expensive screening processes. Here, a methodology for the rapid screening of antibody extraction conditions using a microfluidic channel-based toolbox is presented...
August 5, 2017: Journal of Chromatography. A
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