In vitro antioxidant and in vivo xanthine oxidase inhibitory activities of Pandanus amaryllifolius in potassium oxonate‐induced hyperuricemic rats

Xanthine oxidase (XO) plays an important role in the regulation of uric acid and prevents it from being overproduced as in hyperuricemia disease. The combined effects of antioxidant and xanthine oxidase inhibitor would become a promising approach for hyperuricemia treatment. In this research, the antioxidant and xanthine oxidase inhibitory activities of Pandanus amaryllifolius leaf were evaluated. The leaf water extract (PA‐W) showed highest total phenols, and petroleum ether extract (PA‐PE) showed highest total flavonoids contents. The antioxidant activity of DPPH, metal chelating and hydrogen peroxide was highest in PA‐W extract. The treatment of PA‐W extract at 1000 mg kg^?1 body weight in potassium oxonate‐induced hyperuricemic rats showed significant (P < 0.001) decrease in serum uric acid level by 85% and XO activity by 64%, respectively, as compared to the hyperuricemic rats. In conclusion, the P. amaryllifolius possess the dual effect of antioxidant and XO inhibition as potential therapeutic agents in the hyperuricemia treatment.     

大麦、麦芽和啤酒酿造中的内源性氧化还原酶

由于大麦和麦芽中存在的内潭性氧化还原酶的作用，使内源性多酚、不饱和脂类等物质被氧化，导致成品啤酒风味稳定性和非生物稳定性的降低。酶促氧化反应可发生在不同的酿造阶段，包括发芽、焙爆和糖化等环节。五种主要的内源性氧化还原酶中，超氧化物歧化酶是最为重要的抗氧化酶，可防止超氧阴离子自由基的危害；而过氧化氢酶可催化具有活性的H2O2生成如，由此构成了清除活性氧的初级抗氧化酶体系。过氧化物酶和多酚氧化酶分别在H2O2和O2存在的情况下，可催化内源性酚类底物生成具有活性的醌类物质，所产生的次级氧化产物可改变啤酒的品质。脂肪酸氧化酶可氧化不饱和脂肪酸生成可挥发性的醛类物质，是导致啤酒风味老化的关键酶。酶促氧化的结果在成品啤酒上主要表现为老化异昧的出现、形成混浊、苦味和收敛性的改变，以及色泽的加深等。本文综述了这五种酶的基本性质，在制麦和糖化过程中的变化及其影响，并探讨了对啤酒酿造的影响。     

Structural Data on the Periplasmic Aldehyde Oxidoreductase PaoABC from Escherichia coli: SAXS and Preliminary X-ray Crystallography Analysis

The periplasmic aldehyde oxidoreductase PaoABC from Escherichia coli is a molybdenum enzyme involved in detoxification of aldehydes in the cell. It is an example of an αβγ heterotrimeric enzyme of the xanthine oxidase family of enzymes which does not dimerize via its molybdenum cofactor binding domain. In order to structurally characterize PaoABC, X-ray crystallography and small angle X-ray scattering (SAXS) have been carried out. The protein crystallizes in the presence of 20% (w/v) polyethylene glycol 3350 using the hanging-drop vapour diffusion method. Although crystals were initially twinned, several experiments were done to overcome twinning and lowering the crystallization temperature (293 K to 277 K) was the solution to the problem. The non-twinned crystals used to solve the structure diffract X-rays to beyond 1.80 Å and belong to the C2 space group, with cell parameters a = 109.42 Å, b = 78.08 Å, c = 151.77 Å, β = 99.77°, and one molecule in the asymmetric unit. A molecular replacement solution was found for each subunit separately, using several proteins as search models. SAXS data of PaoABC were also collected showing that, in solution, the protein is also an αβγ heterotrimer.     

Mutations Associated with Functional Disorder of Xanthine Oxidoreductase and Hereditary Xanthinuria in Humans

Xanthine oxidoreductase (XOR) catalyzes the conversion of hypoxanthine to xanthine and xanthine to uric acid with concomitant reduction of either NAD⁺ or O₂. The enzyme is a target of drugs to treat hyperuricemia, gout and reactive oxygen-related diseases. Human diseases associated with genetically determined dysfunction of XOR are termed xanthinuria, because of the excretion of xanthine in urine. Xanthinuria is classified into two subtypes, type I and type II. Type I xanthinuria involves XOR deficiency due to genetic defect of XOR, whereas type II xanthinuria involves dual deficiency of XOR and aldehyde oxidase (AO, a molybdoflavo enzyme similar to XOR) due to genetic defect in the molybdenum cofactor sulfurase. Molybdenum cofactor deficiency is associated with triple deficiency of XOR, AO and sulfite oxidase, due to defective synthesis of molybdopterin, which is a precursor of molybdenum cofactor for all three enzymes. The present review focuses on mutation or chemical modification studies of mammalian XOR, as well as on XOR mutations identified in humans, aimed at understanding the reaction mechanism of XOR and the relevance of mutated XORs as models to estimate the possible side effects of clinical application of XOR inhibitors.     

The WWOX/HIF1A Axis Downregulation Alters Glucose Metabolism and Predispose to Metabolic Disorders

Recent reports indicate that the hypoxia-induced factor (HIF1α) and the Warburg effect play an initiating role in glucotoxicity, which underlies disorders in metabolic diseases. WWOX has been identified as a HIF1α regulator. WWOX downregulation leads to an increased expression of HIF1α target genes encoding glucose transporters and glycolysis’ enzymes. It has been proven in the normoglycemic mice cells and in gestational diabetes patients. The aim of the study was to determine WWOX’s role in glucose metabolism regulation in hyperglycemia and hypoxia to confirm its importance in the development of metabolic disorders. For this purpose, the WWOX gene was silenced in human normal fibroblasts, and then cells were cultured under different sugar and oxygen levels. Thereafter, it was investigated how WWOX silencing alters the genes and proteins expression profile of glucose transporters and glycolysis pathway enzymes, and their activity. In normoxia normoglycemia, higher glycolysis genes expression, their activity, and the lactate concentration were observed in WWOX KO fibroblasts in comparison to control cells. In normoxia hyperglycemia, it was observed a decrease of insulin-dependent glucose uptake and a further increase of lactate. It likely intensifies hyperglycemia condition, which deepen the glucose toxic effect. Then, in hypoxia hyperglycemia, WWOX KO caused weaker glucose uptake and elevated lactate production. In conclusion, the WWOX/HIF1A axis downregulation alters glucose metabolism and probably predispose to metabolic disorders.     

The Periplasmic Oxidoreductase DsbA Is Required for Virulence of the Phytopathogen Dickeya solani

In bacteria, the DsbA oxidoreductase is a crucial factor responsible for the introduction of disulfide bonds to extracytoplasmic proteins, which include important virulence factors. A lack of proper disulfide bonds frequently leads to instability and/or loss of protein function; therefore, improper disulfide bonding may lead to avirulent phenotypes. The importance of the DsbA function in phytopathogens has not been extensively studied yet. Dickeya solani is a bacterium from the Soft Rot Pectobacteriaceae family which is responsible for very high economic losses mainly in potato. In this work, we constructed a D. solani dsbA mutant and demonstrated that a lack of DsbA caused a loss of virulence. The mutant bacteria showed lower activities of secreted virulence determinants and were unable to develop disease symptoms in a potato plant. The SWATH-MS-based proteomic analysis revealed that the dsbA mutation led to multifaceted effects in the D. solani cells, including not only lower levels of secreted virulence factors, but also the induction of stress responses. Finally, the outer membrane barrier seemed to be disturbed by the mutation. Our results clearly demonstrate that the function played by the DsbA oxidoreductase is crucial for D. solani virulence, and a lack of DsbA significantly disturbs cellular physiology.     

Syntheses of 1,2,4-triazolo[4,3-c]pteridines. Potential substrates for xanthine oxidase

A synthetic route for the preparation of 9-phenyl-1,2,4-triazolo[4,3-c]pteridines 8 is described. Their reactivities towards xanthine oxidase from Arthrobacter M-4 are determined and compared with the pteridine derivatives 6-phenylpteridin-4[3H]-thione 5, 6-phenyl-4-thiomethylpteridine 6, 4-hydrazino-6-phenylpteridine 7, 3-N-methyl-6-phenylpteridin-4[3H]-thione 19, 4-amino-6-phenylpteridine 22 and 1-N-methyl-6-phenylpteridin-4[1H]-one 24, in order to inspect a possible influence of pyrimidine ring substitution pattern on their interaction with the bacterial enzyme. It has been suggested that a planar structure of the pteridine moiety and the substituents could be an important factor that so far has been overlooked when considering the enzyme activity. Some literature data are also discussed in view of revision of structure assignment of the compounds 12, 14, and 17. This revised version was published online in August 2006 with corrections to the Cover Date.     

Probes and nano-delivery systems targeting NAD(P)H:quinone oxidoreductase 1: a mini-review

The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues. NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers. Therefore, it could be a target for cancer therapy. Because it is a quinone reductase, many bioimaging probes based on quinone structures target NAD(P)H:quinone oxidoreductase 1 to diagnose tumours. Its expression is higher in tumours than in normal tissues, and using target drugs such as β-lapachone to reduce side effects in normal tissues can help. However, the physicochemical properties of β-lapachone limit its application. The problem can be solved by using nanosystems to deliver β-lapachone. This mini-review summarizes quinone-based fluorescent, near-infrared and two-photon fluorescent probes, as well as nanosystems for delivering the NAD(P)H:quinone oxidoreductase 1-activating drug β-lapachone. This review provides valuable information for the future development of probes and nano-delivery systems that target NAD(P)H:quinone oxidoreductase 1.     

Cloning and heterologous expression of the NADPH cytochrome P450 oxidoreductase genes from an industrial dicarboxylic acid-producing Candida tropicalis

NADPH cytochrome P450 oxidoreductase (CPR) catalyses the transfer of electrons during P450-mediated oxidation, which plays an important role in the omega-oxidation pathway of Candida tropicalis. Two putative allelic genes, CPR-a and CPR-b, were cloned from the long chain dicarboxylic acid-producing Candida tropicalis 1230, using cassette PCR methods. Both the identified open reading frames predict the gene products of 679 amino acid residues. The deduced amino acid sequences of CPR-a and CPR-b are highly homologous to CPR genes from C. tropicalis ATCC 750 and Candida maltosa. Both genes were individually expressed in a cpr mutant of Saccharomyces cerevisiae with high CPR activities, in which only a small distinction was observed between recombinant CPR-a and CPR-b. Both CPR-a and CPR-b contain one CTG codon, which codes for serine (amino acid 50) in C. tropicalis rather than universal leucine. A mutated cDNA of CPR-a with a TCG codon instead of CTG codon was constructed and expressed, resulting in little increase in CPR activity. This indicates that the alteration of Ser-50 has little effect on functional expression of CPR. Furthermore, high ketoconazole sensitivity for the cpr mutant was complemented by heterologous expression of the cloned CPR-a or CPR-b.     

Antioxidative activities of bran extracts from twenty one pigmented rice cultivars

Ethanol–water (70:30, v/v) extracts from the bran of rice seeds from twenty one pigmented and one nonpigmented rice cultivars were evaluated for antioxidative activities using the following tests: inhibition of peroxidation of linoleic acid; inhibition of peroxidation of rabbit lipid erythrocyte membranes; reduction of potassium ferricyanide, and scavenging of superoxide anions and hydroxyl radicals. With some exceptions, extracts from the pigmented rice seeds had higher antioxidative activity than did the nonpigmented variety. The following pigmented cultivars had the highest antioxidative activities in all tests: Jumlalocal-1, Parnkhari 203, DZ78, LK1-3-6-12-1-1, and Elwee. A significant correlation was also noted between reducing power, inhibition of erythrocyte ghost membrane peroxidation, and superoxide anion and hydroxyl radical scavenging. The results suggest that: (a) ferricyanide reducing power might be a useful and simple index for large-scale evaluation of antioxidative potencies of natural products present in rice; (b) pigmented rice varieties with high antioxidative activities provide a source of antioxidants and a genetic resource to develop new health-promoting rice cultivars.