scholarly journals Determining a Critical Threshold for G6PD Activity below which RBC Response to Oxidative Stress is Poor

2020 ◽  
Author(s):  
Maria Swastika ◽  
Alida R Harahap ◽  
Lydia V Panggalo ◽  
Sri Widia A Jusman ◽  
Ari W Satyagraha
Keyword(s):  
Oxidative Stress ◽  
G6pd Deficiency ◽  
Ex Vivo ◽  
Strong Association ◽  
G6pd Activity ◽  
World Health ◽  
Critical Threshold ◽  
Main Function ◽  
Oxidative Markers ◽  
Significant Difference

Abstract Background Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme disorder in the world. Its main function is to generate NADPH that is required for anti-oxidative pathway in the cells especially in red blood cells (RBC). G6PD deficiency is X-linked and thus subject to random X-chromosome inactivation in women giving them mosaic expression of G6PD activities in their individual cells. This phenomenon makes it difficult for diagnosis with the currently available G6PD qualitative diagnostic tests. With the rolling out of newly marketed anti-malarial drug tafenoquine, which has a long half-life, screening for G6PD deficiency becomes a necessity where those with <70% G6PD activity cannot receive this drug. Thus, evidence for a quantitative cut-off for G6PD activity is needed to ensure safe drug administration.Methods RBC models were developed to analyse the effect of oxidant on RBC oxidative markers namely total glutathione (GSH)and malondialdehyde (MDA). G6PD activity was measured using quantitative assay from Trinity Biotech and was correlated with cytofluorometric assay. RBC from twoG6PD heterozygous women with different G6PD activities were also analysed for comparison.Results There was a negative correlation between G6PD activity and CuCl concentration and a strong association between G6PD activities and proportion of G6PD normal RBC in CuCl-treated models and in ex vivo RBC. However, in terms of oxidative stress markers analyses, unlike the hypothesis where the lower G6PD activity, the higher MDA and the lower GSH level, the CuCl RBC model showed that in low G6PD activities (10-30%) cells, the MDA level is lower compared to the rest of the models (p<0.05). The ex vivo models however were in line with the hypothesis, although the result was not significant (p=0.5). There was a significant difference between RBC with <60% and those with >80% G6PD activities in CuCl RBC model, but not in ex vivo RBC (p=0.5). Genotyping heterozygous subjects showed G6PDViangchan variant with 2.97U/gHb (33% activity) and 6.58 U/gHb (74% activity). Conclusions The GSH analysis has pointed to the 60% G6PD activity cut-off and this data is supportive of the old World Health Organization threshold for intermediate upper limit of 60% G6PD activity. However, there are significant limitations in using MDA assay with CuCl RBC model because the RBC was already stressed due to the copper treatment and thus present a different result when compared to the ex-vivo model.

scholarly journals Determining a Critical Threshold for G6PD Activity below which RBC Response to Oxidative Stress is Poor

2020 ◽  
Author(s):  
Maria Swastika ◽  
Alida R Harahap ◽  
Lydia V Panggalo ◽  
Sri Widia A Jusman ◽  
Ari W Satyagraha
Keyword(s):  
Oxidative Stress ◽  
G6pd Deficiency ◽  
Ex Vivo ◽  
Strong Association ◽  
G6pd Activity ◽  
Critical Threshold ◽  
Main Function ◽  
Ex Vivo Model ◽  
Comparison Results ◽  
Significant Difference

Abstract Background: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme disorder in the world. Its main function is to generate NADPH that is required for anti-oxidative pathway in the cells especially in RBC. G6PD deficiency is X-linked and thus subject to random X-chromosome inactivation in women giving them mosaic expression of G6PD activities in their individual cells. This phenomenon makes it difficult for diagnosis with the currently available G6PD qualitative diagnostic tests. With the rolling out of newly marketed anti-malarial drug tafenoquine which has a long half-life, screening for G6PD deficiency becomes a necessity where those with <70% G6PD activity cannot receive this drug. Thus, evidence for a quantitative cut-off for G6PD activity is needed to ensure safe drug administration. Methods: RBC models were developed to analyze the effect of oxidant on RBC oxidative markers namely total glutathione (GSH) and malondialdehyde (MDA). G6PD activity was measured using quantitative assay from Trinity Biotech and was correlated with cytofluorometric assay. RBC from two G6PD heterozygous women with different G6PD activities were also analyzed for comparison. Results: There was a negative correlation between G6PD activity and CuCl concentration and a strong association between G6PD activities and proportion of G6PD normal RBC in CuCl-treated models and in ex vivo RBC. However, in terms of oxidative stress markers analyses, unlike the hypothesis where the lower G6PD activity, the higher MDA and the lower GSH level, our CuCl RBC model showed that in low G6PD activities (10-30%) cells, the MDA level is lower compared to the rest of the models (p<0.05). Our ex vivo model however were in line with the hypothesis, although the result was not significant (p=0.5). There was a significant difference between RBC with <60% and those with >80% G6PD activities in CuCl RBC model but not in ex vivo RBC (p=0.5). Genotyping heterozygous subjects showed G6PD Viangchan variant with 2.97 U/gHb (33% activity) and 6.58 U/gHb (74% activity). Conclusions: The GSH analysis has pointed to the 60% G6PD activity cut-off and this data is supportive of the old WHO threshold for intermediate upper limit of 60% G6PD activity. However, there are significant limitations in using MDA assay with CuCl RBC model because the RBC was already stressed due to the copper treatment and thus present a different result when compared to the ex-vivo model.

scholarly journals Determining Quantitative Diagnostic Test Cut-off in G6PD Heterozygous Women for Tafenoquine Therapy

2019 ◽  
Author(s):  
Maria Swastika ◽  
Alida R Harahap ◽  
Lydia V Panggalo ◽  
Sri Widia A Jusman ◽  
Ari W Satyagraha
Keyword(s):  
G6pd Deficiency ◽  
Ex Vivo ◽  
Strong Association ◽  
G6pd Activity ◽  
Main Function ◽  
Ex Vivo Model ◽  
Stress Markers ◽  
Oxidative Markers ◽  
Significant Difference ◽  
Glucose 6 Phosphate Dehydrogenase

Abstract Background Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme disorder in the world. Its main function is to generate NADPH that is required for anti-oxidative pathway in the cells especially in red blood cells (RBC). G6PD deficiency is X-linked and thus subject to random X-chromosome inactivation in women giving them mosaic expression of G6PD activities in their individual cells. This phenomenon makes it difficult for diagnosis with the currently available G6PD qualitative diagnostic tests. With the rolling out of newly marketed anti-malarial drug tafenoquine which has a long half-life, screening for G6PD deficiency becomes a necessity where those with <70% G6PD activity cannot receive this drug. Thus, evidence for a quantitative cut-off for G6PD activity is needed to ensure safe drug administration.Methods RBC models were developed to analyze the effect of oxidant on RBC oxidative markers namely total glutathione (GSH) and malondialdehyde (MDA). G6PD activity was measured using quantitative assay from Trinity Biotech and was correlated with cytofluorometric assay. RBC from G6PD heterozygous women with different G6PD activities were also analyzed for comparison.Results There was a negative correlation between G6PD activity and CuCl concentration and a strong association between G6PD activities and proportion of G6PD normal RBC in CuCl-treated models and in ex vivo RBC. However, in terms of oxidative stress markers analyses, unlike the hypothesis where the lower G6PD activity, the higher MDA and the lower GSH level, our CuCl RBC model showed that in low G6PD activities (10-30%) cells, the MDA level is lower compared to the rest of the models (p<0.05). Our ex vivo model however were in line with the hypothesis, although the result was not significant (p=0.5). There was a significant difference between RBC with <60% and those with >80% G6PD activities in CuCl RBC model but not in ex vivo RBC (p=0.5). Genotyping heterozygous subjects showed G6PD Viangchan variant with 2.97 U/g Hb (33% activity) and 6.58 U/g Hb (74% activity).Conclusions The MDA and GSH analyses have pointed to the 60% G6PD activity cut-off. This provides an evidence of possible cut-off for tafenoquine administration in G6PD heterozygous women.

scholarly journals Glucose-6-Phosphate Dehydrogenase Deficiency among Children Attending the Emergency Department of Yankin Children’s Hospital, Yangon

2019 ◽  
Vol 31 (2) ◽  
pp. 143-147
Keyword(s):  
Emergency Department ◽  
G6pd Deficiency ◽  
Who Classification ◽  
G6pd Activity ◽  
World Health ◽  
In Vitro Test ◽  
Children's Hospital ◽  
Severe Deficiency ◽  
Test Kit ◽  
Children’S Hospital

Yankin Children’s Hospital is one of the tertiary children hospitals in Myanmar, where some oxidative medications are commonly used in the management of illnesses. Paediatrician’s awareness of G6PD status in this population is very important for effective management and prevention of complications in G6PD deficient children. This preliminary study aims to determine the prevalence of G6PD deficiency according to WHO classification among children seeking medical care at Emergency Department of Yankin Children’s Hospital (YKCH). It was a cross-sectional descriptive study on 124 children, aged 1 month to 13 years. G6PD enzyme activity was determined by spectrophotometric assay within 24 hours of sample collection. Randox G6PD quantitative in vitro test kit (Randox Laboratories, Crumlin, UK) was used and G6PD activity was calculated as unit per gram (U/g) of haemoglobin (Hb). For classification of G6PD deficiency, 10% and 60% level of normal enzyme activities were calculated according to the suggestion by World Health Organization (WHO); G6PD activity <10% was defined as severe deficiency and 10-60% was defined as moderate deficiency. According to WHO classification, 18.5% (23/124) of children in this study was classified as G6PD deficient, with 3.2% severe deficiency and 15.3% moderate deficiency. The prevalence of G6PD deficiency in Myanmar children is higher than the previous reported prevalence if quantitative spectrophotometric method is used for diagnosis, detecting more individuals with moderate deficiency. The high prevalence of G6PD deficiency in this study warrants for the need to do neonatal screening to avoid the potentially fatal complications of this disease.

scholarly journals PO 8609 PREVALENCE AND RISK FACTORS FOR GLUCOSE-6-PHOSPHATE DEHYDROGENASE (G6PD) DEFICIENCY IN TWO P. VIVAX MALARIA-ENDEMIC AREAS IN SUDAN

2019 ◽  
Vol 4 (Suppl 3) ◽  
pp. A62.2-A62
Author(s):  
Muzamil Mahdi Abdel Hamid ◽  
Musab Albsheer ◽  
Mohamed Muneer ◽  
Lina Altinae ◽  
Andrew A Lover
Keyword(s):  
Risk Factors ◽  
G6pd Deficiency ◽  
Demographic Data ◽  
Univariate Analysis ◽  
G6pd Activity ◽  
Radical Cure ◽  
Major Health Problem ◽  
Significant Difference ◽  
Endemic Areas ◽  
Glucose 6 Phosphate Dehydrogenase

BackgroundPlasmodium vivax malaria is a major health problem in Sudan and the parasite has become widely distributed in the recent years. The WHO recommends the use of primaquine as radical cure for liver dormant stage, the hypnozoite. However, prior its use, a test for Glucose-6-phosphate Dehydrogenase (G6PD) should be performed. The objective of the current study was to determine prevalence and risk factors for G6PD deficiency in two P. vivax malaria-endemic areas in Sudan.MethodsA cross-sectional study recruiting 557 subjects from two malaria-endemic areas in Sudan was conducted. Demographic data and blood samples were collected. G6PD activity was measured by spectrometry using SPINREACT enzymatic-UV kit.ResultsThe measured G6PD activities for both sites ranged from 0.6 to 37.7 U/g Hb, with a median value of 12.8 U/g Hb. There was a significant difference in enzyme activity by study site (p<0.001), but not by sex (p=0.91). Overall, across the two study sites, 22 (3.9%) is G6PDd (<30%). Prevalence of G6PDd (<30%) in Khartoum is 1.8% (4/230) compared to 4.8% (16/327) in New Hafla. In univariate analysis predictors of G6PDd were study site (odds ratio of G6PD activity <3.8, Khartoum relative to New Halfa=0.22 (95% CI: 0.08 to 0.66), p=0.006), and recent antibiotic use (OR=2.45 (95% CI: 1.1 to 5.5), p=0.027). In multivariate analysis, the only factor that was significant was the individual’s weight in kilograms, with an OR of 0.97 (95% CI 0.95 to 0.99, p=0.014).ConclusionG6PD deficiency is less prevalent among Sudanese population and this indicates that the use of primaquine for radical cure of P. vivax malaria is safe.

Frequency of Human Paraoxonase-1 Q192R Polymorphism and Measurement of Oxidative Stress Parameters in Infants with G6PD Deficiency

2020 ◽  
Author(s):  
Vahid Pourshafiei ◽  
Vahide Jamshidi ◽  
Ameneh Khodarahmi ◽  
Mahmood Vakili
Keyword(s):  
Oxidative Stress ◽  
G6pd Deficiency ◽  
Paraoxonase 1 ◽  
Pon1 Activity ◽  
Stress Markers ◽  
Genotype Frequencies ◽  
Q192r Polymorphism ◽  
Significant Difference ◽  
And Control

Background and Aims: This study aimed to investigate the frequency of Q192R polymorphism and oxidative stress markers in infants with glucose-6phosphate dehydrogenase (G6PD) deficiency. Materials and Methods: This is a case-control study in which 60 male infants (2-4 months old) with G6PD deficiency along with 60 age- and sexmatched healthy neonates were included. The diagnosis of G6PD deficiency was made by Beutler test by which the G6PD enzyme activity is measured by the fluorescent spot test. The blood samples were taken from all infants, and the sera were isolated for the evaluation of Paraoxonase-1 (PON1) and malondialdehyde (MDA) using the spectrophotometric method. Restriction fragment length polymorphism was applied for determination of Q192R polymorphism (rs 662). Results: The frequencies of QQ, QR, and RR genotypes were 55%, 39%, and 6%, respectively in infants with G6PD deficiency while the above genotype frequencies were 45%, 49%, and 6%, respectively in healthy neonates. The frequency of R and T alleles failed to show any significant difference when G6PD deficient infants and healthy neonates were compared. The results indicated PON1 activity and MDA levels being significantly (p<0.05) higher in neonates with G6PD deficiency compared with their healthy counterparts. Conclusion: Contrary to previous studies, it was indicated that the presence of RQ and RR genotypes at Q192R position is associated with decreased activity of PON1 and increased oxidative stress. In this study, no significant differences were found in the genotype and allele frequency of PON1 Q192R polymorphism between the case and control groups. Also, this frequency was not consistent with the results obtained from oxidative stress conditions.

scholarly journals Determining a critical threshold for G6PD activity below which red blood cell response to oxidative stress is poor

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Maria Swastika ◽  
Alida R. Harahap ◽  
Lydia V. Panggalo ◽  
Sri Widia A. Jusman ◽  
Ari W. Satyagraha
Keyword(s):  
Oxidative Stress ◽  
Blood Cell ◽  
Red Blood Cell ◽  
Cell Response ◽  
G6pd Activity ◽  
Critical Threshold

scholarly journals G6PD deficiency is associated with renal impairment in T2DM

2020 ◽  
Author(s):  
Nabin Bahadur Basnet ◽  
Santosh Pradhan ◽  
Rishi Kumar Kafle ◽  
Keyoor Gautam
Keyword(s):  
Oxidative Stress ◽  
High Risk ◽  
Renal Impairment ◽  
Diabetic Patient ◽  
G6pd Deficiency ◽  
G6pd Activity ◽  
Diabetic Patients ◽  
Healthy Control ◽  
Disease Study ◽  
Severe Degree

Abstract Background Diabetes and its complications including renal impairment are the consequences of increased oxidative stress. Glucose-6-phosphate dehydrogenase (G6PD) is responsible for protection from harmful oxidative metabolites. G6PD deficiency hence leaves these individuals at high risk of oxidative stress and at high risk of diabetes and its complications including renal impairment. If any association between G6PD deficiency and damage to kidney exists in diabetic population then screening for G6PD deficiency in this population will help identify the susceptible patient who may be at higher risk of diabetic complications due to G6PD deficiency and can prevent them with early interventions to help decrease patients’ morbidity in the long run. Methods Healthy control volunteers and diabetic with or without renal impairment were selected after counseling. Body Mass Index and blood pressure were recorded. Blood tests to measure the G6PD activity in fresh whole blood, hemoglobin, glycated hemoglobin, uric acid, and creatinine were done. Urine was tested for the presence of proteinuria. The activity of G6PD was measured. Glomerular filtration rate (eGFR) was estimated with the Modification of Diet in Renal Disease Study (MDRD) Equation. Calculations and statistical analysis were done with open software LibreOffice 6.3.3 and SPSS version 22.0.Results The mean G6PD level of healthy control was 84.27 ± 9 and that of diabetic patient without renal impairment and diabetic patients with renal impairment were 65.05 ± 5.35 and 46.64 ± 6.56 respectively. There was only one participant (4.55%) with G6PD deficiency among healthy controls whereas the proportion of participants having normal, mild to moderate, and severe degree of G6PD deficiency were 77.27%, 13.64%, and 9.09%, respectively for diabetic patient without renal impairment while 40.91%, 22.73%, and 36.36% for those with renal impairment. The proportion of participants with renal impairment had more severe degree of G6PD deficiencies than those without renal impairment (p=0.03).Conclusion The prevalence of G6PD deficiency was higher in diabetics with renal impairment than those with diabetes alone suggesting the relation between G6PD activity and diabetes with renal impairment.

scholarly journals Ex Vivo Study of Laban's Role in Decreasing Hemolysis Crisis in G6PD-Deficient Patients

2020 ◽  
Vol 2020 ◽  
pp. 1-5 ◽  
Author(s):  
Wissam Zam ◽  
Loay Belal
Keyword(s):  
Human Blood ◽  
G6pd Deficiency ◽  
Ex Vivo ◽  
Antioxidant Properties ◽  
G6pd Activity ◽  
Environmental Benefits ◽  
Fermented Dairy Products ◽  
Fava Beans ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Fermented Dairy

In spite of the vast nutritional and environmental benefits provided by fava bean (Vicia faba), the ingestion of vicine/convicine provokes an acute hemolytic anemia called favism in individuals with a glucose-6-phosphate dehydrogenase (G6PD) deficiency. The elimination of these glycosides is a goal that could be accomplished using different processing methods including bacteriological treatment. Laban as a good source of lactic acid bacteria was tested in an ex vivo assay on human blood samples in order to determine its capacity in decreasing the hemolysis crisis induced by the ingestion of fava beans. Results indicate a significant decrease in human blood cell hemolysis after the treatment of fava beans by Laban. This decrease in hemolysis was also correlated with the G6PD deficiency categorization. The highest hemolysis level (mean: 23.11 ± 0.76%) was observed in samples with G6PD activity between 10 and 30%, while the lowest hemolysis level (mean: 5.75 ± 0.64%) was observed in samples with G6PD activity more than 60%. This decrease was correlated with a high antioxidant capacity of Laban (51.61 ± 1.13% expressed by the percentage inhibition of DPPH radical). The counts of isolates from MRS and M17 culture plates were 6.75 ± 0.095 and 7.91 ± 0.061 log cfu ml–1, respectively. In conclusion, the synergy between the antioxidant properties of Laban and the possible decrease of vicine and convicine concentrations by lactobacillus found in the fermented dairy products could explain the ability of Laban to reduce the hemolysis crisis ex vivo.

scholarly journals The Frequency of PON1 L55M Polymorphism and Measurement of Oxidative Stress Parameters (Superoxide Dismutase, Catalase, Thiol, and Carbonyl) in Neonates with G6PD Deficiency Compared to the Control Group

2020 ◽  
Author(s):  
Vahide Jamshidi ◽  
Vahid Pourshafi ◽  
Mahmoud Vakili ◽  
Ali Moradi
Keyword(s):  
Oxidative Stress ◽  
Superoxide Dismutase ◽  
Enzyme Activity ◽  
G6pd Deficiency ◽  
Control Group ◽  
Case Group ◽  
Genotypic Frequency ◽  
Oxidative Stress Parameters ◽  
Significant Difference ◽  
Stress Parameters

Introduction: Glucose-6-phosphate dehydrogenase deficiency is the most common enzyme disorder. This enzyme involved in maintaining the balance of active oxygen species and its defect causes oxidative damage. PON1 is an HDL-based glycosylated protein that prevents lipid peroxidation. In this study, the prevalence of PON L55M polymorphism in paraoxonase enzyme in neonates with a deficiency in G6PD activity was evaluated, and the level of oxidative stress was measured. Methods:  In the present case-control study, 60 infants 2 to 6 months with G6PD enzyme activity deficiency and 60 healthy infants identical in age was selected. Polymorphism examination was done using PCR-RFLP technique, and oxidative stress parameters were measured by spectrophotometry. Chi square and t test statistical analysis of data was performed using SPSS V16 software. Results: The frequency of genotype LL, LM and MM for PON1 L55M was 43.33%, 43.3% and 13.3% and, 35%, 21.6% and 43.3% in control and case group, respectively. Genotypic frequency of LM and MM was significant between control and control groups (P <0.05). The allele frequency between L and M was also significant (P <0.05). Superoxide dismutase enzyme activity and mean carbonyl level comparison did not show a significant difference (P >0.05), but the activity of catalase enzyme and mean level of thiol was showed a significant difference (P <0.05). Conclusion: In the present study, the frequency of LM genotype in neonates with G6PD deficiency was significantly different in comparison to the control group. This frequency is consistent with the results obtained from oxidative stress conditions (significant reduction in the level of thiol and catalase activity).

Sign in / Sign up

Export Citation Format

Share Document