Activity Of G6PD Research Articles

Cyclosporine A alleviates colitis by inhibiting the formation of neutrophil extracellular traps via the regulating pentose phosphate pathway

BackgroundThe aberrant formation of neutrophil extracellular traps (NETs) has been implicated in ulcerative colitis (UC), a chronic recurrent intestinal inflammation. Cyclosporine A (CsA) is now applied as rescue therapy for acute severe UC. In addition, it has been certained that CsA inhibits the formation of NETs in vitro and the mechanism of which was still vague. The study aimed to explore the mechanism CsA inhibits the NETs formation of colitis in vivo and in vitro.MethodsNETs enrichment in clinical samples was analyzed using databases from Gene Expression Omnibus and verified in our center. Dextran sulfate sodium (DSS)-induced acute colitis mice model was used to investigate the effect of CsA on NETs of colonic tissue expression. To clarify the mechanism, intracellular energy metabolites were examined by Liquid Chromatograph Mass Spectrometer, and reactive oxygen species (ROS) levels were examined by fluorescence intensity in neutrophils treated with CsA after LPS stimulation. The transcriptional level and activity of G6PD of neutrophils were also assessed using qRT-PCR and WST-8. RNA Sequencing was used to detect differentially expressed genes of neutrophils stimulated by LPS with or without CsA. The expression levels of related proteins were detected by western blot.ResultsNETs enrichment was especially elevated in moderate-to-severe UC patients compared to HC. NETs expression in the colon from DSS colitis was decreased after CsA treatment. Compared with neutrophils stimulated by LPS, NETs formation and cellular ROS levels were decreased in LPS + CsA group. Cellular ribulose 5-phosphate and NADPH/NADP + related to the pentose phosphate pathway (PPP) were reduced in LPS + CsA group. In addition, CsA could decrease G6PD activity in neutrophils stimulated with LPS, and the results were further verified by inhibiting G6PD activity. At last, P53 protein was highly expressed in LPS + CsA group compared with the LPS group. Intracellular G6PD activity, ROS level and NETs formation, which were downregulated by CsA, could be reversed by a P53 inhibitor.ConclusionOur results indicated CsA could alleviate the severity of colitis by decreasing the formation of NETs in vivo. In vitro, CsA reduced ROS-dependent NETs release via downregulating PPP and cellular ROS levels by decreasing G6PD activity directly by activating the P53 protein.

TIM-3/Gal-9 interaction affects glucose and lipid metabolism in acute myeloid leukemia cell lines.

T-cell immunoglobulin and mucin domain-3 (TIM-3) is a transmembrane molecule first identified as an immunoregulator. This molecule is also expressed on leukemic cells in acute myeloid leukemia and master cell survival and proliferation. In this study, we aimed to explore the effect of TIM-3 interaction with its ligand galectin-9 (Gal-9) on glucose and lipid metabolism in AML cell lines. HL-60 and THP-1 cell lines, representing M3 and M5 AML subtypes, respectively, were cultured under appropriate conditions. The expression of TIM-3 on the cell surface was ascertained by flow cytometric assay. We used real-time PCR to examine the mRNA expression of GLUT-1, HK-2, PFKFB-3, G6PD, ACC-1, ATGL, and CPT-1A; colorimetric assays to measure the concentration of glucose, lactate, GSH, and the enzymatic activity of G6PD; MTT assay to determine cellular proliferation; and gas chromatography-mass spectrometry (GC-MS) to designate FFAs. We observed the significant upregulated expression of GLUT-1, HK-2, PFKFB-3, ACC-1, CPT-1A, and G6PD and the enzymatic activity of G6PD in a time-dependent manner in the presence of Gal-9 compared to the PMA and control groups in both HL-60 and THP-1 cell lines (p > 0.05). Moreover, the elevation of extracellular free fatty acids, glucose consumption, lactate release, the concentration of cellular glutathione (GSH) and cell proliferation were significantly higher in the presence of Gal-9 compared to the PMA and control groups in both cell lines (p < 0.05). TIM-3/Gal-9 ligation on AML cell lines results in aerobic glycolysis and altered lipid metabolism and also protects cells from oxidative stress, all in favor of leukemic cell survival and proliferation.

A GSTP1-mediated lactic acid signaling promotes tumorigenesis through the PPP oxidative branch

Lactic acidosis is a feature of solid tumors and plays fundamental role(s) rendering cancer cells to adapt to diverse metabolic stresses, but the mechanism underlying its roles in redox homeostasis remains elusive. Here we show that G6PD is phosphorylated at tyrosine 249/322 by the SRC through the formation of a GSTP1-G6PD-SRC complex. Lactic acid attenuates this formation and the phosphorylation of G6PD by non-covalently binding with GSTP1. Furthermore, lactic acid increases the activity of G6PD and facilitates the PPP (NADPH production) through its sensor GSTP1, thereby exhibiting resistance to reactive oxygen species when glucose is scarce. Abrogating a GSTP1-mediated lactic acid signaling showed attenuated tumor growth and reduced resistance to ROS in breast cancer cells. Importantly, positive correlations between immuno-enriched SRC protein and G6PD Y249/322 phosphorylation specifically manifest in ER/PR positive or HER negative types of breast cancer. Taken together, these results suggest that GSTP1 plays a key role in tumor development by functioning as a novel lactate sensor.

Glucose-6-Phosphate Dehydrogenase Deficiency in Icteric Newborns at the Essos-Yaound&amp;#233;-Cameroon Hospital

Background: Glucose-6-phosphate dehydrogenase deficiency is an enzymopathy characterized by insufficient production of reduced glutathione (GSH), a molecule known for its antioxidant role. This lack of GSH leads to a deficit in the elimination of peroxide ions from the red blood cells, causing thereby hemolytic accidents, which can be fatal if not properly managed. In neonates, the clinical picture is most often that of neonatal jaundice. Objectives: This study aimed to determine the place of G6PD deficiency as a cause of neonatal jaundice at Essos Hospital Centre. Methods: We conducted a prospective descriptive study over three months. Blood samples taken from newborns aged 0 to 28 days were analyzed in the medical analysis laboratory of the Essos Hospital Centre in Yaoundé. We carried out a determination of the enzymatic activity of G6PD, a blood count and the determination of the bilirubin level. The results obtained were analysed using R statistical software version 4.1.1. Linear regression analyses were used to assess correlations between the variables of interest. Results: Sixty-nine icteric neonates constituted our study population, with a total of 40 boys (58%) and 29 girls (42%) with a sex ratio of 1.37 in favour of boys. The prevalence of G6PD deficiency in icteric children was 50.72%. The mean hemoglobin was 15.3 ± 3.08 g/dL and the mean red blood cell count was 4.52 ± 1.01 × 106/mm3. The mean total bilirubin was 122 ± 48.3 mg/L with a maximum of 308 mg/L and the mean free bilirubin was 104 ± 46.6 mg/L with a maximum of 292 mg/L. Furthermore, after linear regression analysis, we obtained a positive and significant correlation between G6PD enzymatic activity and hemoglobin level (r = 0.33; p ≤ 0.001), G6PD red blood cell level (r = 0.26; p Conclusion: Neonatal jaundice in G6PD-deficient children is a real public health problem and the prevention of hemolysis in children requires an early diagnosis of the enzyme disorder and good follow-up of the children.

Mitochondrial Generated Redox Stress Differently Affects the Endoplasmic Reticulum of Circulating Lymphocytes and Monocytes in Treatment-Naïve Hodgkin’s Lymphoma

Background. The redox stress caused by Hodgkin’s lymphoma (HL) also involves the peripheral blood mononucleated cells (PBMCs) even before chemotherapy. Here, we tested whether lymphocytes and monocytes show a different response to the increased mitochondrial generation of reactive oxygen species (ROS). Methods. PBMCs, isolated from the blood of treatment-naïve HL patients and control subjects, underwent assessment of malondialdehyde content and enzymatic activity of both hexose- and glucose-6P dehydrogenase (H6PD and G6PD) as well as flow cytometric analysis of mitochondrial ROS content. These data were complemented by evaluating the uptake of the fluorescent glucose analogue 2-NBDG that is selectively stored within the endoplasmic reticulum (ER). Results. Malondialdehyde content was increased in the whole population of HL PBMCs. The oxidative damage matched an increased activity of G6PD, and even more of H6PD, that trigger the cytosolic and ER pentose phosphate pathways, respectively. At flow cytometry, the number of recovered viable cells was selectively decreased in HL lymphocytes that also showed a more pronounced increase in mitochondrial ROS generation and 2-NBDG uptake, with respect to monocytes. Conclusions. PBMCs of HL patients display a selective mitochondrial and ER redox stress most evident in lymphocytes already before the exposure to chemotherapy toxicity.

Co-inheritance of G6PD deficiency and 211\u2009G to a variation of UGT1A1 in neonates with hyperbilirubinemia in eastern Guangdong

BackgroundGlucose-6-phosphate dehydrogenase (G6PD) deficiency, which may manifest as neonatal hyperbilirubinemia, is the most prevalent erythrocytic enzyme-related disease in the world.ObjectiveTo investigate the association between neonatal hyperbilirubinemia and co-inheritance of G6PD deficiency and 211 G to A variation of UGT1A1 in Chaozhou city of eastern Guangdong province, the effects of G6PD deficiency and UGT1A1 gene variant on the bilirubin level were determined in neonates with hyperbilirubinemia.MethodThe activity of G6PD was assayed by an auto-bioanalyzer. PCR and flow-through hybridization were used to detect 14 common G6PD mutations in G6PD deficient neonates. 211 G to A variation of UGT1A1 was determined by PCR and sequencing. The data of neonatal bilirubin was collected and analyzed retrospectively.ResultsSeventy four cases of the 882 hyperbilirubinemia neonates were G6PD deficiency (8.39%) while 12 cases of the 585 non-hyperbilirubinemia neonates (control group) were G6PD deficiency (2.05%). The rate of G6PD deficiency in the hyperbilirubinemia group was higher than that of the control group. Moreover, the peak bilirubinin of the G6PD-deficient group of hyperbilirubinemia neonates was 334.43 ± 79.27 μmol/L, higher than that of the normal G6PD group of hyperbilirubinemia neonates (300.30 ± 68.62 μmol/L). The most common genotypes of G6PD deficiency were c.1376G > T and c.1388G > A, and the peak bilirubin of neonates with these two variants were 312.60 ± 71.81 μmol/L and 367.88 ± 75.79 μmol/L, respectively. The bilirubin level of c.1388G > A was significantly higher than that of c.1376G > T. Among the 74 hyperbilirubinemia neonates with G6PD deficiency, 6 cases were 211 G to A homozygous mutation (bilirubin levels 369.55 ± 84.51 μmol/L), 27 cases were 211 G to A heterozygous mutation (bilirubin levels 341.50 ± 63.21 μmol/L), and 41 cases were wild genotypes (bilirubin levels 324.63 ± 57.52 μmol/L).ConclusionThe rate of G6PD deficiency in hyperbilirubinemia neonates was significantly higher than that of the non-hyperbilirubinemia neonates in Chaozhou. For the hyperbilirubinemia group, neonates with G6PD deficiency had a higher bilirubin level compared to those with normal G6PD. For hyperbilirubinemia neonates with G6PD deficiency, there was a declining trend of bilirubin levels among 211 G to A homozygous mutation, heterozygous mutation, and wild genotype, but there was no significance statistically among the three groups.

Imaging of metabolic activity adaptations to UV stress, drugs and differentiation at cellular resolution in skin and skin equivalents – Implications for oxidative UV damage

The epidermis is a multi-layered epithelium that consists mainly of keratinocytes which proliferate in its basal layer and then differentiate to form the stratum corneum, the skin's ultimate barrier to the environment. During differentiation keratinocyte function, chemical composition, physical properties, metabolism and secretion are profoundly changed. Extrinsic or intrinsic stressors, like ultraviolet (UV) radiation thus may differently affect the epidermal keratinocytes, depending on differentiation stage. Exposure to UV elicits the DNA damage responses, activation of pathways which detoxify or repair damage or induction of programmed cell death when the damage was irreparable. Recently, rapid diversion of glucose flux into the pentose phosphate pathway (PPP) was discovered as additional mechanism by which cells rapidly generate reduction equivalents and precursors for nucleotides – both being in demand after UV damage. There is however little known about the correlation of such metabolic activity with differentiation state, cell damage and tissue localization of epidermal cells. We developed a method to correlate the activity of G6PD, the first and rate-limiting enzyme of this metabolic UV response, at cellular resolution to cell type, differentiation state, and cell damage in human skin and in organotypic reconstructed epidermis. We thereby could verify rapid activation of G6PD as an immediate UVB response not only in basal but also in differentiating epidermal keratinocytes and found increased activity in cells which initiated DNA damage responses. When keratinocytes had been UVB irradiated before organotypic culture, their distribution within the skin equivalent was abnormal and the G6PD activity was reduced compared to neighboring cells. Finally, we found that the anti-diabetic and potential anti-aging drug metformin strongly induced G6PD activity throughout reconstructed epidermis. Activation of the protective pentose phosphate pathway may be useful to enhance the skin's antioxidant defense systems and DNA damage repair capacity on demand.

Effect of Statins (Zocor) on Oxygen-Dependent Processes in Muscle Tissue and Erythrocytes in Animals with Hypercholesterolemia

Background. Statin therapy may be accompanied by a toxic effect on skeletal muscle and liver cells, the molecular mechanisms of which have not yet been fully understood.Aim.To clarify the peculiarities of changes in the oxygen transport function of erythrocytes and antioxidant mechanisms in erythrocytes and muscles of animals with prolonged administration of statins (simvastatin).Materials and methods.The study was conducted on outbred male rats, which during the experiment were randomly assigned to following groups: the control group contained animals on a common ration of the vivarium, the group of comparison – animals with induced hypercholesterolemia, the main group – animals with induced hypercholesterolemia treated with simvastatin. In erythrocytes and muscle tissue of animals, indicators characterizing the state of antioxidant defense and carbohydrate metabolism were determined.The results.The experiment showed that the administration of simvastatin to animals was characterized by aggravation of hypoxia due to hypercholesterolemia, as indicated by a sharp increase in the concentration of 2,3-BPG and lactate in erythrocytes of animals, as well as a significant decrease in the activity of G6PD. In the muscle tissue of animals, there was a decrease in the concentration of pyruvic acid and lactate relative to the comparison group, which indicates their intensive participation in metabolic processes.Conclusion.Based on the data obtained, it can be assumed that the peculiarity of the action of statins in erythrocytes is the increased oxygen delivery to the tissues, which in muscles was accompanied by a decrease in the level of oxidized products. At the same time, against the background of the use of statins, despite the positive direction of adaptive reactions, signs of oxidative stress remain, which is documented by the imbalance of the SOD-catalase system and a decrease in the activity of glutathione-dependent reactions

Purification and determination the molecular weight and study the kinetic properties of Glucose-6-phosphate dehydrogenase from Diabetic patients

This study was conducted to purifity Glucose 6-phosphate dehydrogenase from diabetic patients by gel filtration technique on Sephadex G100 and determine the molecular weight of enzyme, concentration and kinetic constant [Km, Vmax], study the effect of optimum temperature, substrate, pH. The study includes [60] patients with diabetes mellitus and [60] healthy individual as control. The activity of G6PD was measured after precipitated by [75%] Ammonium Sulfate concentration, using [Tris-HCl] buffer at pH 8.2. The specific activity was calculated [21.5UI\mg], total activity [706.8UI], number of purification [3.45] enzyme yield [23.188%] and enzyme activity [17.67UI\ml]. and the molecular weight were [57.82] k.D. The effect of substrate concentration was found to be increased with increasing substrate concentration. The optimum pH was found to be [8.4] and the optimum temperature was [38C]. Michaelis- Menten [Km] value for the enzyme was [3.33mM] and Vmax value was [0.263IU\ml ].

The synthesis of new oxindoles as analogs of natural product 3,3′-bis(indolyl)oxindole and in vitro evaluation of the enzyme activity of G6PD and 6PGD

Natural and synthetic derivatives that contain an indole core are being used in medical treatments and technological processes. Therefore, the development of new synthetic methods for the synthesis of indole derivatives is very popular. In this study, new oxindoles with reaction of 4,7-dihydro-1$H$-indole (2) and isatin (4) were synthesized as analogs of natural product 3,3$^\prime $-bis(indolyl)oxindole. The biological properties of the compounds obtained during this study were also studied, showing that compounds 5, 7, and 12 inhibited the activity of G6PD with an IC$_{50}$ of 99 $\mu $M, 231 $\mu $M, and 304 $\mu $M respectively. The activity of rat erythrocyte 6PGD was increased in the presence of 5 and 7 and was inhibited in the presence of 12. As indole derivative 5 was an activator of 6PGD and inhibitor of G6PD, it was selected for docking studies to understand the mechanism of activation and inhibition.

Expression of g6pd gene in wild type zebrafish embryos of early development

: To observe the expression of g6pd gene in the early development stage of wild zebrafish embryos. : The collinearity of g6pd gene and the sequence similarity of G6pd protein were analyzed with gene database and BLAST software, respectively. Expression of g6pd gene in different development stages of zebrafish embryos was detected by in situ hybridization. The g6pd-EGFP-pCS2+ recombinant plasmids were microinjected into zebrafish embryos, and fluorescence was observed under a fluorescence microscope. The expression of G6pd protein at 24, 48 and 72 hour post fertilization (hpf) zebrafish embryos was detected by Western blotting; the enzyme activity of G6pd at 24, 48 and 72 hpf zebrafish embryos was detected by modified G6pd quantitative ratio method. : The G6pd protein similarity of zebrafish and human was 88%, and that of zebrafish and mouse was 87%. The results of in situ hybridization showed that the g6pd gene was mainly expressed in the hematopoietic tissues of zebrafish; the results observed after microinjection of g6pd-EGFP-pCS2+ recombinant plasmid were consistent with the results of in situ hybridization. At 24, 48 and 72 hpf, the relative expression levels of G6pd protein in zebrafish embryos were 1.44±0.03, 1.47±0.05, and 1.54±0.02, respectively(P>0.05); the G6pd enzyme activity levels were 1.74±0.17, 1.75±0.12, 1.71±0.22, respectively (P>0.05). : The study has observed the expression of g6pd gene and G6pd protein, and G6pd enzyme activity in zebrafish embryos at different development phases, which provides a reference for the establishment of a zebrafish G6PD deficiency model.

Purification, Determination Molecular Weight and Study Kinetic Properties of G6PD from Diabetes Patient

This study was conducted to purification G6PD enzyme from diabetic patients by using simple and cheap method the technique gel filtration on Sephadex G100 and determine molecular weight of enzyme and compare it with true molecular weight of enzyme and determine kinetic constant (Km, Vmax) and study the effect of temperature and substrate and pH and known the best condition to give optimum work of enzyme. study contain (60) patients with diabetes and (60) control Glucose and activity of G6PD were measured and the enzyme precipitated by Ammonium Sulfate with concentration (75%) and purification enzyme gel filtration on Sephadex G-100 with dimensions (1.5 × 30) cm and using the buffer solution from (Tris-HCl) at pH 8.2 to isolate the enzyme and determine molecular weight with same method. Specific activity was calculated (21.5 UI/mg), total activity (706.8 UI), number of purification (3.45) enzyme yield (23.188%) and enzyme activity (17.67 UI/ml). and the molecular weight was calculated with using same technique (57.82) kD. Effect of increased concentration of substrate on enzyme activity and found the activity increase with increase substrate and amount constant level not change however increase of concentration of substrate when drawing relation between activity and concentration of substrate format appear exchange excess and after study effect of pH found the optimum value (8.4) and study effect of temperature on activity found (38 C) the optimum temperature. Study of kinetic constant was done and the and the Michaelis-Menten (Km) value was (3.8 mM) and Vmax value (8 IU/ml).

Erythrocyte Oxidative Stress Markers in Children: A Clinical Laboratory Experience

Because oxidative damage to erythrocytes plays an important pathophysiologic role in many diseases and there are some difficulties in obtaining reference intervals for children in many laboratory tests, this work intends to describe the determination of oxidative stress parameters and the results obtained for children. A total of 280 blood samples were obtained from children between 8 and 11 years old, without any hematological disease. Samples were analyzed for seven oxidative stress parameters, methemoglobin, reduced glutathione, TBARS, percentage of hemolysis and activity of the enzymes G6PD, superoxide dismutase, and catalase. To draw a comparison, the same parameters were analyzed in children with sickle cell disease. Because all results presented normal distribution in the Kolmogorov-Smirnov, data were expressed as 2.5 and 97.5th accumulated percentiles and the statistical analysis between male and female children was performed using Student t -test. A p value

Purification and characterization of glucose-6-phosphate dehydrogenase from camel liver.

Glucose-6-phosphate dehydrogenase from camel liver was purified to homogeneity by ammonium sulfate precipitation and a combination of DEAE-cellulose, Sephacryl S-300 gel filtration, and 2′, 5′ ADP Sepharose 4B affinity chromatography columns. The specific activity of camel liver G6PD is increased to 1.80438 units/mg proteins with 63-fold purification. It turned out to be homogenous on both native PAGE and 12% SDS PAGE, with a molecular weight of 64 kDa. The molecular weight of the native form of camel liver G6PD was determined to be 194 kDa by gel filtration indicating a trimeric protein. The K m value was found to be 0.081 mM of NADP+. Camel liver G6PD displayed its optimum activity at pH 7.8 with an isoelectric point (pI) of pH 6.6–6.8. The divalent cations MgCl2, MnCl2, and CoCl2 act as activators; on the other hand, CaCl2 and NiCl2 act as moderate inhibitors, while FeCl2, CuCl2, and ZnCl2 are potent inhibitors of camel liver G6PD activity. NADPH inhibited camel liver G6PD competitively with K i value of 0.035 mM. One binding site was deduced for NADPH on the enzyme molecule. This study presents a simple and reproducible purification procedure of G6PD from the camel liver.

Abstract 2433: Metabolic reprogramming by an epigenetic mechanism in endocrine therapy resistance of breast cancer

Abstract Metabolic reprogramming is a major mechanism of fueling cancer cell growth and proliferation. However, how tumor metabolism is altered to confer therapeutic resistance is poorly understood. Gene expression profiling of tamoxifen sensitive and resistant breast cancer cells revealed that transition to the resistance was associated with a specific expression reprogramming of genes involved in glucose/energy metabolism, which was accompanied by aberration of specific epigenetic regulators such as the histone methyltransferase NSD2/MMSET/WHSC1. In cell and xenograft tumor models, NSD2 overexpression alone was sufficient to confer tamoxifen resistance. Analysis of clinical specimens indicates that NSD2 was a strong predictive factor for early relapse of tamoxifen therapy. NSD2 wild type, but not its methylase-defective mutant form, coordinately stimulated the expression and enzymatic activity of HK2, TIGAR and G6PD and strongly augmented the pentose phosphate pathway (PPP) production of NADPH for ROS reduction and survival of tamoxifen treated cells and tumors. Further mechanistic studies demonstrated that elevated NSD2 stimulates the metabolic gene expression through H3K36 methylation at the target chromatin. Pharmacological targeting aberrant NSD2 by DZNeP, an S-adenosylhomocysteine (AdoHcy) hydrolase inhibitor, effectively blocked the growth of tamoxifen-resistant tumor. The study reveals a novel mechanism of cancer metabolic reprogramming and suggests targeting the key metabolic re-programmer as a viable option for effective treatment of endocrine resistant tumors. Note: This abstract was not presented at the meeting. Citation Format: June X. Zou, Junjian Wang, Zhijian Duan, Hongwu Chen, Hsing-Jien Kung, Xinbin Chen, Leigh C. Murphy, Alexander Borowsky. Metabolic reprogramming by an epigenetic mechanism in endocrine therapy resistance of breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 2433. doi:10.1158/1538-7445.AM2014-2433

PHYSIOLOGICAL AND BIOCHEMICAL STUDY FOR SPECIMEN OF CHILDREN SUFFERING OF ENZYME G6PD DEFICIENCY AT RAMADI CITY

Some blood variable and reduced Glutathione (GSH) concentration had been studied besides lipid peroxides (Malondialdehyde (MDA)) and the concentration of trace ions of the element (Fe+2 , Cu+2 , Zn+2) in the blood serum of ill mace children. The study including sixty children suffering of acute hemolytic anemia of ages between (1-10)years ,more over another 30 healthy childs appearance of the some ages as a control group . The result indicated significant decrease at (P ≤ 0.05) of the activity of the enzyme G6PD in the children suffering illness due to the enzymatic deficiency in comparison with healthy children where the value of the enzyme activity recorded illustrated that the suffered children recorded (1.77 ± 0.71) U/g.Hb while the healthy children was (12.64 ± 2.40) U/g.Hb . The result also showed the children suffering G6PD lack recorded low significant result (P ≤ 0.05) at both the concentration of hemoglobin , site of packed red blood cells , reduced Glutathione and the concentration of Zn+2 relatively to healthy, where its concentration for the suffering children (5.82 ± 1.16) g/dl , (18.43 ± 3.81) % , (2.95 ± 0.99) µmol/l and (15.78 ± 4.23) µmol/l while for the healthy were (11.95 ± 0.67) g/dl , (37.76 ± 1.98) % , (7.80 ± 2.20) µmol/l , (20.60±3.06) µmol/l respectively. While it revealed significant elevation (P ≤ 0.05) at both white blood cells and the MDA concentration ,besides the concentration of Fe+2 and Cu+2 at the Infected children regarding the healthy children. Where by its concentration at Infected children (8.46 ± 2.03)*103 cell/mm3 , (20.83 ± 7.75) µmol/l , (81.75 ± 48.58) µmol/l and (44.29 ± 10.88) µmol/l. while for healthy were (16.14 ± 5.57)*103 cell/mm3 , (8.98 ± 2.47) µmol/l , (33.48 ± 5.23) µmol/l and (33.19 ± 6.76) µmol/l. respectively.

Stimulation of gene expression and activity of antioxidant related enzyme in Sprague Dawley rat kidney induced by long-term iron toxicity

The trace elements such as iron are vital for various enzyme activities and for other cellular proteins, but iron toxicity causes the production of reactive oxygen species (ROS) that causes alterations in morphology and function of the nephron. The present study was designed to determine the effect of long-term iron overload on the renal antioxidant system and to determine any possible correlation between enzymatic and molecular levels. Our data showed that reduced glutathione (GSH) levels, which is a marker for oxidative stress, strikingly decreased with a long-term iron overload in rat kidney. While renal mRNA levels of glucose 6-phosphate dehydrogenase (G6pd), 6-phosphogluconate dehydrogenase (6pgd) and glutathione peroxidase (Gpx) were significantly affected in the presence of ferric iron, no changes were seen for glutathione reductase (Gsr) and glutathione S-transferases (Gst). While the iron affected the enzymatic activity of G6PD, GSR, GST, and GPX, it had no significant effect on 6PGD activity in the rat kidney. In conclusion, we reported here that the gene expression of G6pd, 6pgd, Gsr, Gpx, and Gst did not correlate to enzyme activity, and the actual effect of long-term iron overload on renal antioxidant system is observed at protein level. Furthermore, the influence of iron on the renal antioxidant system is different from its effect on the hepatic antioxidant system.

PHYSIOLOGICAL AND BIOCHEMICAL STUDY FOR SPECIMEN OF CHILDREN SUFFERING OF ENZYME G6PD DEFICIENCY AT RAMADY CITY.

Some blood variable and reduced Glutathione (GSH) concentration had been studied besides lipid peroxides (Malondialdehyde (MDA)) and the concentration of trace ions of the element (Fe+2 , Cu+2 , Zn+2) in the blood serum of ill mace children. The study including sixty children suffering of acute hemolytic anemia of ages between (1-10)years ,more over another 30 healthy childs appearance of the some ages as a control group .The result indicated significant decrease at (P ≤ 0.05) of the activity of the enzyme G6PD in the children suffering illness due to the enzymatic deficiency in comparison with healthy children where the value of the enzyme activity recorded illustrated that the suffered children recorded (1.77 ± 0.71) U/g.Hb while the healthy children was (12.64 ± 2.40) U/g.Hb . The result also showed the children suffering G6PD lack recorded low significant result (P ≤ 0.05) at both the concentration of hemoglobin , site of packed red blood cells , reduced Glutathione and the concentration of Zn+2 relatively to healthy, where its concentration for the suffering children (5.82 ± 1.16) g/dl , (18.43 ± 3.81) % , (2.95 ± 0.99) µmol/l and (15.78 ± 4.23) µmol/l while for the healthy were (11.95 ± 0.67) g/dl , (37.76 ± 1.98) % , (7.80 ± 2.20) µmol/l , (20.60±3.06) µmol/l respectively. While it revealed significant elevation (P ≤ 0.05) at both white blood cells and the MDA concentration ,besides the concentration of Fe+2 and Cu+2 at the Infected children regarding the healthy children. Where by its concentration at Infected children (8.46 ± 2.03)*103 cell/mm3 , (20.83 ± 7.75) µmol/l , (81.75 ± 48.58) µmol/l and (44.29 ± 10.88) µmol/l. while for healthy were (16.14 ± 5.57)*103 cell/mm3 , (8.98 ± 2.47) µmol/l , (33.48 ± 5.23) µmol/l and (33.19 ± 6.76) µmol/l. Respectively.

Effect of Hibiscus sabdariffa anthocyanins on 2, 4- dinitrophenylhydrazine-induced tissue damage in rabbits

This study examines the effects of anthocyanin extract of the dried calyces of Hibiscus sabdariffa Linn. on the 2, 4 -dinitrophenylhydrazine (2, 4 -DNPH)-induced cytotoxic effects in rabbits. Twenty male adult rabbits used for the study were divided into four groups. Group 1, the control took only water while animals in groups 2 and 4 received 100 mg/kg body weight of the anthocyanin extract of H. sabdariffa once daily for 28 days. After the 22nd day of treatment, the rabbits in groups 3 and 4 received 28 mg/kg body weight of DNPH for the remaining 5 days of treatment, after which the animals were sacrificed. Relative to control, DNPH caused significant (p < 0.05) increase in the formation of malondialdehyde (MDA) in serum, liver and brain and decreased the levels of reduced glutathione (GSH) in liver and brain. Also, DNPH caused a significant (p < 0.05) elevation in the activity of glucose- 6 -phosphate dehydrogenase (G6PD) in the serum and liver. However, pretreatment with H. sabdariffa anthocyanin extract significantly (p < 0.05) reduced MDA formation, increased the levels of GSH and maintained at normalcy the activity of G6PD in the tissues, thereby effectively ameliorated the toxic effects of DNPH. These findings indicate that anthocyanin extract from dried calyces of H. sabdariffa protects the rabbit against 2, 4 -DNPH lipoperoxidative and cytotoxic effects. Key words: Anthocyanin extract, 2, 4-dinitrophenylhydrazine, glucose-6-phosphate dehydrogenase, Hibiscus sabdariffa, rabbit, reduced glutathione, malondialdehyde

Exercise induced hemolysis: relation between the activity of glucose-6-phosphate dehydrogenase and the magnitude of the hemolysis.

Introduction: Oxidation and hemolysis associated with physical exercise, moderate and severe, have been studied by several investigations, but only in the most recent have been evaluated how the variations of the genetic determined mechanisms for antioxidative enzymatic erytrocytic capacity, contribute to establish different levels of susceptibility to the hemolysis induced trough exercise, however, those studies have mainly been made on non sedentary subjects with deficiency of enzyme G6PD. Purpose: The objective of this study was to establish if there is either yes or not a relationship between the activity level of erythrocytic activity of the enzyme G6PD and the hemolysis grade, induced by exercise in adult healthy sedentary men and after an experimental bout of moderate exercise pre-exhaustive and equivalent in modality and intensity. Methods: Twenty five men, sedentary and seemingly healthy, were evaluated after an exercise session of 1 hour over a cycloergometer, with 70% of VO2peak previously founded. On the basis of plasmatic Hb, the proportion of hemolysis and haptoglobin consumed, taking in account the variation of plasma haptoglobin and intravascular hemolysis after and at least three hours post-exercise, were calculated. Results: The experimental subjects presented laboratory evidence (free plasma hemoglobin, percentage of hemolysis and haptoglobin) of intravascular hemolysis immediately after and at least past three hours of exercise bouts. Conclusion: The grade of hemolysis had an inversely significative correlation with the G6PD activity and, this intensity had a significative different behaviour below the percentil 40 media founded for G6PD and above of it. Thus, the oxidative hemolytic response in sedentary and healthy individuals who had lower levels G6PD activity with one bout of exercise with 70% of VO2peak was highly variable.