Search results for: G6PD

Authors: Haiyoung Jung, Mi Joung Leem, Sun Hwa Lee

Abstract:

Background & Objective: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is a genetic abnormality that results in an inadequate amount of G6PD, leading to increased susceptibility of red blood cells to reactive oxygen species and hemolysis. The present study aimed to evaluate the careSTARTTM S1 analyzer for measuring G6PD activity to hemoglobin (Hb) ratio. Methods: Precision for G6PD activity and hemoglobin measurement was evaluated using control materials with two levels on five repeated runs per day for five days. The analytic performance of the careSTARTTM S1 analyzer was compared with spectrophotometry in 40 patient samples. Reference ranges suggested by the manufacturer were validated in 20 healthy males and females each. Results: The careSTARTTM S1 analyzer demonstrated precision of 6.0% for low-level (14~45 U/dL) and 2.7% for high-level (60~90 U/dL) control in G6PD activity, and 1.4% in hemoglobin (7.9~16.3 u/g Hb). A comparison study of G6PD to Hb ratio between the careSTARTTM S1 analyzer and spectrophotometry showed an average difference of 29.1% with a positive bias of the careSTARTTM S1 analyzer. All normal samples from the healthy population were validated for the suggested reference range for males (≥2.19 U/g Hb) and females (≥5.83 U/g Hb). Conclusion: The careSTARTTM S1 analyzer demonstrated good analytical performance and can replace the current spectrophotometric measurement of G6PD enzyme activity. In the aspect of the management of clinical laboratories, it can be a reasonable option as a point-of-care analyzer with minimal handling of samples and reagents, in addition to the automatic calculation of the ratio of measured G6PD activity and Hb concentration, to minimize any clerical errors involved with manual calculation.

Keywords: POCT, G6PD, performance evaluation, careSTART

Procedia PDF Downloads 39

Authors: Ludmila A. Gavriliuc, Jerry McLarty, Heather E. Kleiner, J. Michael Mathis

Abstract:

Abstract -- Background: The citrus coumarine Auraptene (Aur) is an effective chemopreventive agent, as manifested in many models of diseases and cancer. Nuclear factor erythroid 2-related factor (Nrf2) is an important regulator of genes induced by oxidative stress, such as glutathione S-transferases, heme oxygenase-1, and peroxiredoxin 1, by activating the antioxidant response element (ARE). Genetic and biochemical evidence has demonstrated that glutathione (GSH) and glutathione-dependent enzymes, glutathione reductase (GR), glutathione peroxidases (GPs), glutathione S-transferases (GSTs) are responsible for the control of intracellular reduction-oxidation status and participate in cellular adaptation to oxidative stress. The effect of Aur on the activity of GR, GPs (Se-GP and Se-iGP), and content of GSH in the liver, kidney, and spleen is insufficiently explored. Aim: Our goal was the examination of the Aur influence on the redox-system of GSH in Nrf2 wild type and Nrf2 knockout mice via activation of Nrf2 and ARE. Methods: Twenty female mice, 10 Nrf2 wild-type (WT) and 10 Nrf2 (-/-) knockout (KO), were bred and genotyped for our study. The activity of GR, Se-GP, Se-iGP, GST, G6PD, CytP450 reductase, catalase (Cat), and content of GSH were analyzed in the liver, kidney, and spleen using Spectrophotometry methods. The results of the specific activity of enzymes and the amount of GSH were analyzed with ANOVA and Spearman statistical methods. Results: Aur (200 mg/kg) treatment induced hepatic GST, GR, Se-GP activity and inhibited their activity in the spleen of mice, most likely via activation of the ARE through Nrf2. Activation in kidney Se-GP and G6PD by Aur is also controlled, apparently through Nrf2. Results of the non-parametric Spearman correlation analysis indicated the strong positive correlation between GR and G6PD only in the liver in WT control mice (r=+0.972; p < 0.005) and in the kidney KO control mice (r=+0.958; p < 0.005). The observed low content of GSH in the liver of KO mice indicated an increase in its participation in the neutralization of toxic substances with the absence of induction of GSH-dependent enzymes, such as GST, GR, Se-GP, and Se-iGP. Activation of CytP450 in kidney and spleen and Cat in the liver in KO mice probably revealed another regulatory mechanism for these enzymes. Conclusion: Thereby, obtained results testify that Aur can modulate the activity of genes and antioxidant enzymatic redox-system of GSH, responsible for the control of intracellular reduction-oxidation status.

Keywords: auraptene, glutathione, GST, Nrf2

Procedia PDF Downloads 109

Authors: Antonella Di Sotto, Cinzia Ingallina, Caterina Fraschetti, Simone Circi, Marcello Locatelli, Simone Carradori, Gabriela Mazzanti, Luisa Mannina, Silvia Di Giacomo

Abstract:

Cannabis sativa L. is one of the oldest cultivated plant species known not only for its voluptuous use but also for the wide application in food, textile, and therapeutic industries. Recently, the progress of biotechnologies applied to medicinal plants has allowed to produce different hemp varieties with low content of psychotropic phytoconstituents (tetrahydrocannabinol < 0.2% w/v), thus leading to a renewed industrial and therapeutic interest for this plant. In this context, in order to discover new potential remedies of pharmaceutical and/or nutraceutical interest, the chemopreventive properties of different organic and hydroalcoholic extracts, obtained from the inflorescences of C. sativa L. var. USO31, collected in June and September harvesting, were assessed. Particularly, the antimutagenic activity towards the oxidative DNA-damage induced by tert-butyl hydroperoxide (t-BOOH) was evaluated, and the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid) radical scavenging power of the samples were assessed as possible mechanisms of antimutagenicity. Furthermore, the ability of the extracts to inhibit the glucose-6-phosphate dehydrogenase (G6PD), whose overexpression has been found to play a critical role in neoplastic transformation and tumor progression, has been studied as a possible chemopreventive strategy. A careful phytochemical characterization of the extracts for phenolic and terpenoid composition has been obtained by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) methods. Under our experimental condition, all the extracts were found able to interfere with the tBOOH-induced mutagenicity in WP2uvrAR strain, although with different potency and effectiveness. The organic extracts from both the harvesting periods were found to be the main effective antimutagenic samples, reaching about a 55% inhibition of the tBOOH-mutagenicity at the highest concentration tested (250 μg/ml). All the extracts exhibited radical scavenger activity against DPPH and ABTS radicals, with a higher potency of the hydroalcoholic samples. The organic extracts were also able to inhibit the G6PD enzyme, being the samples from September harvesting the highly potent (about 50% inhibition respect to the vehicle). At the phytochemical analysis, all the extracts resulted to contain both polar and apolar phenolic compounds. The HPLC analysis revealed the presence of catechin and rutin as the major constituents of the hydroalcoholic extracts, with lower levels of quercetin and ferulic acid. The monoterpene carvacrol was found to be an ubiquitarian constituent. At GC-MS analysis, different terpenoids, among which caryophyllene sesquiterpenes, were identified. This evidence suggests a possible role of both polyphenols and terpenoids in the chemopreventive properties of the extracts from the inflorescences of C. sativa var. USO31. According to the literature, carvacrol and caryophyllene sesquiterpenes can contribute to the strong antimutagenicity although the role of all the hemp phytocomplex cannot be excluded. In conclusion, present results highlight a possible interest for the inflorescences of C. sativa var. USO31 as source of bioactive molecules and stimulate further studies in order to characterize its possible application for nutraceutical and pharmaceutical purposes.

Keywords: antimutagenicity, glucose-6-phosphate dehydrogenase, hemp inflorescences, nutraceuticals, sesquiterpenes

Procedia PDF Downloads 121

Authors: S. Thangapandiyan, S. Miltonprabu

Abstract:

Fl (Fl) exposure engenders neurodegeneration and induces oxidative stress in the brain. The Neuroprotective role of EGCG on oxidative stress-mediated neurotoxicity in Fl intoxicated rat hippocampus has not yet been explored so far. Hence, the present study is focused on witnessing whether EGCG (40mg/kg) supplementation prevents Fl induced oxidative stress in the brain of rats with special emphasis on the hippocampus. Fl (25mg/kg) intoxication for four weeks in rats showed an increase in Fl concentration along with the decrease the AChE, NP, DA, and 5-HT activity in the brain. The oxidative stress markers (ROS, TBARS, NO, and PC) were significantly increased with decreased enzymatic (SOD, CAT, GPx, GR, GST, and G6PD) and non-enzymatic antioxidants (GSH, TSH, and Vit.C) in Fl intoxicated rat hippocampus. Moreover, Fl intoxicated rats exhibited an intrinsic and extrinsic pathway mediated apoptosis in the hippocampus of rats. Fl intoxication significantly increased the DNA damage as evidenced by increased DNA fragmentation. Furthermore, the toxic impact of Fl on hippocampus was also proved by the immunohistochemical, histological, and ultrastructural studies. Pre-administration of EGCG has significantly protected the Fl induced oxidative stress, biochemical changes, cellular apoptotic, and histological alternations in the hippocampus of rats. In conclusion, EGCG supplementation significantly attenuated the Fl induced oxidative stress mediated neurotoxicity via its free radical scavenging and antioxidant activity.

Keywords: brain, hippocampal, NaF, ROS, EGCG

Procedia PDF Downloads 359

Authors: So Young Kim, Eun-Young Kwon, Bora Choi, Mi Kyeong Yu, Seon Jeong Lee, Myung-Sook Choi

Abstract:

Lemon (Citrus limon) has various beneficial effect. Eriocitrin (eriodictyol 7-rutinoside) is the main ingredient of lemon fruit and is known to have antioxidative effects. However, there has been little research about the effects of eriocitrin on obesity and regulation of lipid profiles levels. In the present study, we investigated the anti-obesity and lipid-lowering effects of eriocitrin in mice fed high-fat diet (HFD). The 4 week-old male C57BL/6 mice were randomly divided into two groups and were fed HFD (20% fat, w/w) and HFD supplemented with eriocitrin (0.005%, w/w, EC) for 16 weeks. Food intake, body weight and white adipose tissue weight (WAT) were measured and plasma free fatty acid (FFA), apolipoprotein (Apo) B100 level and hepatic enzyme activity were analyzed. No differences were shown between the HFD and EC groups in body weight and food intake. However EC supplementation significantly reduced the weights of epididymal, subcutaneous and total WAT. In addition, the levels of plasma FFA and Apo B100 were significantly decreased in the EC group compared with the HFD group. Moreover, the activities of glucose-6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) related to fatty acids synthesis were significantly lower in the EC group than in the HFD group in liver. Therefore, this study indicates that eriocitrin has beneficial effects on adiposity and nonalcholic fatty liver diseases by modulating hepatic lipid-regulating enzyme activities and plasma lipid profile.

Keywords: antiobesity, eriocitrin, high fat diet, lipid lowering

Procedia PDF Downloads 422