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Margret Thorsteinsdottir, Unnur A Thorsteinsdottir, Finnur F Eiriksson, Hrafnhildur L Runolfsdottir, Inger M Sch Agustsdottir, Steinunn Oddsdottir, Baldur B Sigurdsson, Hordur K Hardarson, Nilesh R Kamble, Snorri Th Sigurdsson, Vidar O Edvardsson, Runolfur Palsson
Adenine phosphoribosyltransferase (APRT) deficiency is a hereditary disorder that leads to excessive urinary excretion of 2,8-dihydroxyadenine (DHA), causing nephrolithiasis and chronic kidney disease. Treatment with allopurinol or febuxostat reduces DHA production and attenuates the renal manifestations. Assessment of DHA crystalluria by urine microscopy is used for therapeutic monitoring, but lacks sensitivity. We report a high-throughput assay based on ultra-performance liquid chromatography coupled to tandem mass spectrometry (UPLC-MS/MS) for quantification of urinary DHA...
September 14, 2016: Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Ross D Milton
Electrochemically-active polymers (redox polymers) are useful tools for simultaneous immobilization and electron transfer of enzymes at electrode surfaces, which also serve to increase the localized concentration of the biocatalyst. The properties of the employed redox couple must be compatible with the target biocatalyst from both an electrochemical (potential) and biochemical standpoint. This chapter details the synthesis of a naphthoquinone-functionalized redox polymer (NQ-LPEI) that is used to immobilize and electronically communicate with flavin adenine dinucleotide-dependent glucose dehydrogenase (FAD-GDH), yielding an enzymatic bioanode that is able to deliver large catalytic current densities for glucose oxidation at a relatively low associated potential...
2017: Methods in Molecular Biology
David P Hickey
Enzymatic glucose biosensors and biofuel cells make use of the electrochemical transduction between an oxidoreductase enzyme, such as glucose oxidase (GOx), and an electrode to either quantify the amount of glucose in a solution or generate electrical energy. However, many enzymes including GOx are not able to electrochemically interact with an electrode surface directly, but require an external electrochemical relay to shuttle electrons to the electrode. Ferrocene-modified linear poly(ethylenimine) (Fc-LPEI) redox polymers have been designed to simultaneously immobilize glucose oxidase (GOx) at an electrode and mediate electron transfer from their flavin adenine dinucleotide (FAD) active site to the electrode surface...
2017: Methods in Molecular Biology
Danielle Aberdein, John S Munday, Barbara Gandolfi, Keren E Dittmer, Richard Malik, Dorian J Garrick, Leslie A Lyons
British shorthair (BSH) kittens in multiple litters died as a result of a severe non-neoplastic lymphoproliferative disease that showed many similarities with human autoimmune lymphoproliferative syndrome (ALPS). Human ALPS is caused by inherited defects in FAS-mediated lymphocyte apoptosis and the possibility of similar defects was investigated in BSH cats. The whole genomes of two affected kittens were sequenced and compared to 82 existing cat genomes. Both BSH kittens had homozygous insertions of an adenine within exon 3 of the FAS-ligand gene...
October 21, 2016: Mammalian Genome: Official Journal of the International Mammalian Genome Society
Lily Marsden, Lawrence J Jennings, Samantha Gadd, Min Yu, Elizabeth J Perlman, Mariana M Cajaiba
Metanephric stromal tumors are rare renal stromal tumors that predominantly affect children. They belong to the metanephric family of tumors along with metanephric adenofibroma and metanephric adenoma. The previous documentation of BRAF exon 15 mutations in 88% of metanephric adenomas and in isolated cases of metanephric adenofibroma prompted us to investigate the prevalence of these mutations in metanephric stromal tumors and in other pediatric renal stromal tumors. In this study, 17 metanephric stromal tumors, 22 congenital mesoblastic nephromas and 6 ossifying renal tumors of infancy were selected for BRAF exon 15 testing...
October 18, 2016: Human Pathology
Qin Wang, Hongjun Chen, Yong Li, Huixia Wang, Zhou Nie, Yufang Hu, Shouzhou Yao
We report here a label-free and sensitive electrochemical method for probing Citrate synthase (CS) activity based on detailed investigations into the nucleic acid-mimicking coordination polymer (CP) formed from the coenzyme A (CoA)-Ag(I) repeat units. Our biosensing approach provides an especial and significant detection mechanism: CS can catalyze the essential condensation reaction between acetyl-coenzyme A (Ac-CoA) and oxaloacetate (OAA) to form citrate and CoA; then, in the presence of Ag(I), CoA-Ag(I) CP can be in situ formed because of the strong complexation ability of thiol groups of CoA toward Ag(I)...
December 1, 2016: Talanta
Wenyan Xu, Daniel Kool, Derek K O'Flaherty, Ashley Keating, Lauralicia Sacre, Martin Egli, Anne Marietta Noronha, Christopher James Wilds, Linlin Zhao
DNA interstrand cross-links (ICLs) are cytotoxic DNA lesions derived from reactions of DNA with a number of anti-cancer reagents as well as endogenous bifunctional electrophiles. Deciphering the DNA repair mechanisms of ICLs is important for understanding the toxicity of DNA cross-linking agents and for the development of effective chemotherapies. Previous research has focused on ICLs cross-linked with the N7 and N2 atoms of guanine as well as those formed at the N6 atom of adenine; however, little is known about the mutagenicity of O6-dG-derived ICLs...
October 21, 2016: Chemical Research in Toxicology
Zhen Wang, Eric P Chang, Vern L Schramm
Transition path sampling simulations have proposed that human heart lactate dehydrogenase (LDH) employs protein promoting vibrations (PPVs) on the femtosecond (fs) to picosecond (ps) timescale to promote crossing of the chemical barrier. This chemical barrier involves both hydride and proton transfers to pyruvate to form L-lactate, using reduced nicotinamide adenine dinucleotide (NADH) as the cofactor. Here we report experimental evidence from three types of isotope effect experiments that support coupling of the promoting vibrations to barrier crossing and the coincidence of hydride and proton transfer...
October 21, 2016: Journal of the American Chemical Society
Sheng-Chun Yu, Alexander Dawson, Alyssa C Henderson, Eloise J Lockyer, Emily Read, Gayathri Sritharan, Marjah Ryan, Mara Sgroi, Pok M Ngou, Rosie Woodruff, Ruifeng Zhang, Travis Ren Teen Chia, Yu Liu, Yiyu Xiang, Pietro D Spanu
Efficiency of yeast transformation is determined by the rate of yeast endocytosis. The aim of this study was to investigate the effect of introducing amino acids and other nutrients (inositol, adenine, or p-aminobenzoic acid) in the transformation medium to develop a highly efficient yeast transformation protocol. The target of rapamycin complex 1 (TORC1) kinase signalling complex influences the rate of yeast endocytosis. TORC signaling is induced by amino acids in the media. Here, we found that increasing the concentration of amino acids and other nutrients in the growth media lead to an increase yeast transformation efficiency up to 10(7) CFU per μg plasmid DNA and per 10(8) cells with a 13...
October 20, 2016: Scientific Reports
Ross D Blundell, Simon J Williams, Samantha D M Arras, Jessica L Chitty, Kirsten L Blake, Daniel J Ericsson, Nidhi Tibrewal, Jurgen Rohr, Y Q Andre E Koh, Ulrike Kappler, Avril A B Robertson, Mark S Butler, Matthew A Cooper, Bostjan Kobe, James A Fraser
Opportunistic fungal pathogens such as Cryptococcus neoformans are a growing cause of morbidity and mortality among immunocompromised populations worldwide. To address the current paucity of antifungal therapeutic agents, further research into fungal-specific drug targets is required. Adenylosuccinate synthetase (AdSS) is a crucial enzyme in the adeosine triphosphate (ATP) biosynthetic pathway, catalyzing the formation of adenylosuccinate from inosine monophosphate and aspartate. We have investigated the potential of this enzyme as an antifungal drug target, finding that loss of function results in adenine auxotrophy in C...
September 9, 2016: ACS Infectious Diseases
Antonino Baez, Joseph Shiloach
The damaging effect of high oxygen concentration on growth of Escherichia coli is well established. Over-oxygenation increases the intracellular concentration of reactive oxygen species (ROS), causing the destruction of the [4Fe-4S] cluster of dehydratases and limiting the biosynthesis of both branched-chain amino acids and nicotinamide adenine dinucleotide. A key enzyme that reduces the damaging effect of superoxide is superoxide dismutase (SOD). Its transcriptional regulation is controlled by global transcription regulators that respond to changes in oxygen and iron concentrations and pH...
October 18, 2016: Antonie Van Leeuwenhoek
Ivyna Bong Pau Ni, Ng Ching Ching, Shaik Kamal Fakiruddin, Lim Moon Nian, Zubaidah Zakaria
Multiple myeloma (MM) is a malignancy of B lymphocytes or plasma cells. Our array-based comparative genomic hybridization findings revealed chromosomal gains at 7q22.3 and 1q42.3, where nicotinamide (NAM) phosphoribosyltransferase (NAMPT) and lysosomal trafficking regulator (LYST) genes are localized, respectively. This led us to further study the functions of these genes in myeloma cells. NAMPT is a key enzyme involved in nicotinamide adenine dinucleotide salvage pathway, and it is frequently overexpressed in human cancers...
October 18, 2016: Bosnian Journal of Basic Medical Sciences
Sanad Alonezi, Jonans Tusiimire, Jennifer Wallace, Mark J Dufton, John A Parkinson, Louise C Young, Carol J Clements, Jin Kyu Park, Jong Woon Jeon, Valerie A Ferro, David G Watson
In the present study, liquid chromatography-mass spectrometry (LC-MS) was employed to characterise the metabolic profiles of two human ovarian cancer cell lines A2780 (cisplatin-sensitive) and A2780CR (cisplatin-resistant) in response to their exposure to melittin, a cytotoxic peptide from bee venom. In addition, the metabolomics data were supported by application of Biolog microarray technology to examine the utilisation of carbon sources by the two cell lines. Data extraction with MZmine 2.14 and database searching were applied to provide metabolite lists...
October 13, 2016: Metabolites
Teeradate Kongpichitchoke, Ming-Tzu Chiu, Tzou-Chi Huang, Jue-Liang Hsu
Teas can be classified according to their degree of fermentation, which has been reported to affect both the bioactive components in the teas and their antioxidative activity. In this study, four kinds of commercial Taiwanese tea at different degrees of fermentation, which include green (non-fermented), oolong (semi-fermented), black (fully fermented), and Pu-erh (post-fermented) tea, were profiled for catechin levels by using high performance liquid chromatography (HPLC). The result indicated that the gallic acid content in tea was directly proportional to the degree of fermentation in which the lowest and highest gallic acid content were 1...
October 12, 2016: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Jun Tao
OBJECTIVE: Hyperaldosteronemia exerts adverse effects on vascular endothelium except enhanceing blood pressure; however, the impacts and molecular mechanisms of hyperaldosteronemia on endothelial progenitor cell (EPC)-mediated endothelial repair are yet to be determined. The aim of this study was to investigate the endothelial repair capacity of EPCs from hypertensive patients with primary hyperaldosteronemia (PHA). DESIGN AND METHOD: In vivo endothelial repair capacity of EPCs from PHAs (n = 20), age- and blood pressure-matched essential hypertension patients (n = 20), and age-matched healthy subjects (n = 20) was evaluated by transplantation into nude mice with carotid artery denudation injury...
September 2016: Journal of Hypertension
Risa Ramadhiani, Muhammad Saifur Rohman, Nashi Widodo, Valerinna Y S Putri, Mifetika Lukitasari, Jayarani F Putri, Didik H Utomo
OBJECTIVE: To investigate whether the angiotensinogen (AGT) G-217A polymorphism affects blood pressure response of telmisartan and valsartan in hypertensive patients. DESIGN AND METHOD: A total of 46 primary hypertensive patients, 23 patients received telmisartan 80 mg and 23 patients received valsartan 160 mg, were observed and followed up for 4 months. The AGT G-217A polymorphism was determined by PCR-RFLP and direct sequencing method. Blood pressure response was measured daytime (6 am - 10 pm), night-time (10 pm - 6 am), and 24 hours using twenty-four hours ambulatory blood pressure monitoring (ABPM)...
September 2016: Journal of Hypertension
Pandeeswar Makam, Satyaprasad P Senanayak, Thimmaiah Govindaraju
Reliable and ultrasensitive detection of mercury ions is of paramount importance for toxicology assessment, environmental protection and human health. Herein, we present a novel optoelectronic approach based on nanoachitectonics of small molecule templated DNA system that consists of an adenine (A) conjugated small organic semiconductor (BNA) and deoxyribo-oligothymidine (dTn). This mutually templated dynamic chiral co-assembly system (BNAn-dTn) with tunable chiroptical, morphological and electrical properties is tapped in to enable ultra-sensitive and selective detection of inorganic and organometallic mercury in water...
October 18, 2016: ACS Applied Materials & Interfaces
Asghar Tofighi, Minoo Shirpoor, M H Khadem Ansari, Alireza Shirpoor, Mitra Zerehpoosh
OBJECTIVE: Chronic anabolic androgenic steroid (AAS) consumption increases incidence of cardiovascular abnormalities in athletes and mechanisms underlying those abnormalities continue to be investigated. This study examines whether nandrolone consumption induced cardiac and coronary artery wall abnormalities via oxidative stress. It was also designed to determine whether enforced swimming augmented possible cardiotoxic effects of nandrolone in rat heart. METHODS: Twenty-four male Wistar rats were divided into 3 groups: control, nandrolone, and nandrolone with enforced swimming...
October 12, 2016: Anatolian Journal of Cardiology
Hua Chen, Gang Cao, Dan-Qian Chen, Ming Wang, Nosratola D Vaziri, Zhi-Hao Zhang, Jia-Rong Mao, Xu Bai, Ying-Yong Zhao
Early detection is critical in prevention and treatment of kidney disease. However currently clinical laboratory and histopathological tests do not provide region-specific and accurate biomarkers for early detection of kidney disease. The present study was conducted to identify sensitive biomarkers for early detection and progression of tubulo-interstitial nephropathy in aristolochic acid I-induced rats at weeks 4, 8 and 12. Biomarkers were validated using aristolochic acid nephropathy (AAN) rats at week 24, adenine-induced chronic kidney disease (CKD) rats and CKD patients...
September 28, 2016: Redox Biology
Tony Pham, Katherine A Forrest, KaiJie Chen, Amrit Kumar, Michael J Zaworotko, Brian Space
Molecular simulations of CO2 and H2 sorption were performed in MPM-1-Cl and MPM-1-TIFSIX, two robust molecular porous materials (MPMs) with the empirical formula [Cu2(adenine)4Cl2]Cl2 and [Cu2(adenine)4(TiF6)2], respectively. Recent experimental studies have shown that MPM-1-TIFSIX displayed higher CO2 uptake and isosteric heat of adsorption (Qst) than MPM-1-Cl [Nugent, P. S.; et al. J. Am. Chem. Soc. 2013, 135, 10950-10953.]. This was verified through the simulations executed herein, as the presented simulated CO2 sorption isotherms and Qst values are in very good agreement with the corresponding experimental data for both MPMs...
October 17, 2016: Langmuir: the ACS Journal of Surfaces and Colloids
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