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 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 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.

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 enzymopathy of G6PD deficiency in Jordan: a demographic and biochemical analysis

2018 ◽  
Vol 10 (1) ◽  
pp. 25-32
Author(s):  
Ahmed Al-Imam
Keyword(s):  
G6pd Deficiency ◽  
Liver Enzymes ◽  
Clinical Manifestations ◽  
University Hospital ◽  
Drug Induced ◽  
Mean Values ◽  
Significant Difference ◽  
Wbc Count ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Blood Grouping

Background: G6PD deficiency is an inherited X-linked recessive condition leading to insufficient levels of glucose-6-phosphate dehydrogenase, thus causing hemolytic anaemia under certain circumstances. Materials and Methods: Our study is explorative for cases admitted to Jordan University Hospital. The studied parameters include demographics, clinical manifestations, biochemical markers including Hb level, WBC count, liver enzymes, and blood grouping. Results: Most of the patients were admitted to the emergency unit (53.13%). Individuals who were Rh-positive represented 57.81%, while patients of AB blood group accounted for 75%. The mean values were 4.81 years (age), 29.06 hours (time-to-hospital admission), 38.10 degree Celsius (temperature), 6.11 gm/dl (Hb), 13242.19 (WBC count), 343.20 U/L (S. ALP), and 50.98 IU/L (S. ALT). There was no significant difference between males and females or between favism-induced versus drug-induced hemolytic episodes. AB and Rh positive blood groups are of a protective effect in relation to liver enzymes. Patients who were admitted to the hospital within 24 hours from having clinical manifestations had a better prognosis. Conclusion: This study is the first inferential research on G6PD deficiency from the Middle East to explore cases from one of the largest healthcare centres in Jordan. The role of blood grouping should be investigated prospectively.

Frequency of G6PD Deficiency in General Population at District Bannu, Khyber Pakhtunkhwa, Pakistan

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Muhammad Shoaib Khan ◽  
Arif Ullah ◽  
Sami ul Haq ◽  
Mohammd Shoaib
Keyword(s):  
General Population ◽  
G6pd Deficiency ◽  
Clinical Laboratory ◽  
Human Subjects ◽  
Cross Sectional ◽  
Khyber Pakhtunkhwa ◽  
Significant Difference ◽  
Moderate Anemia ◽  
Males And Females ◽  
Glucose 6 Phosphate Dehydrogenase

Background: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked disease and it is a common enzymaticdisease of RBCs in humans X linked recessive condition are more common in males than females. The said deficiency leads toaffecting >400 million people worldwide Individuals, normally males, with deficient alleles are helpless to neonatal jaundice andintense hemolytic anemia, usually during disease, after treatment with specific medications or subsequent to eating Fava beans..Objective: To measure the frequency of Glucose-6-Phosphate-Dehydrogenase deficiency in general population at district Bannu,Khyber Pakhtunkhwa Pakistan.Material and Methods: This cross-sectional descriptive study was conducted on 500 human subjects, who were referred forG6PD assay, in Samad clinical Laboratory, District Bannu Khyber Pakhtunkhwa, Pakistan, from July 2018 to July 2019. 500ccvenous whole blood was collected in EDTA containing vial, for G6PD Test. (Span Diagnostic S.A.R.L, France). Patients of any Age,Sex & area having fever, hematuria, headache, visible jaundice, family history, malaria and anemia were included in this study,while patients suffering from renal disease, any malignancy & not willing persons were excluded.Results: Out of the total 500 hundred, 370 (74%) were males and females were 130 (26%). Total 64/500 (12.8 %) were G6PDdeficient, with 55 male and 09 were female. Malaria positive with G6PD deficiency were 13/64 (20.31%), with 12 males and onefemale. Statistically significant difference among each group (p= 0.0022) was noted. Mean age of the G6PD deficient persons was(2.8 ± 1.03) years. Anemia was graded as Hemoglobin less than 11.5g/dl was taken as anemia. Severe anemia as Hb < 7 g/dl,moderate anemia as Hb ranged between 7-10 g/dl and mild if Hb ranged between 10-11.5 g/dl.Among 370 males, 86 persons had hemoglobin of less than 11.5 g/dl, 42 had hemoglobin 7-10 g/dl and 7 patients had hemoglobinless than 7 g/dl, only 235 patients had hemoglobin more than 11.5, among 130 females, 33 patients had hemoglobin of less than11.5 g/dl, 17 patients had hemoglobin 7-10 g/dl and 05 patients had hemoglobin less than 7 g/dl, only 75 patients had hemoglobinmore than 11.5 g/dl.Conclusion: This study shows high frequency of G6PD Deficiency in district Bannu Khyber Pakhtunkhwa Pakistan especially veryhigh frequency in males than females.

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 A Tetrazolium-Linked Cytochemical Method for Estimation of Glucose-6-Phosphate Dehydrogenase Activity in Individual Erythrocytes: Applications in the Study of Heterozygotes for Glucose-6-Phosphate Dehydrogenase Deficiency

Blood ◽  
1968 ◽  
Vol 31 (5) ◽  
pp. 589-603 ◽  
Author(s):  
VIRGIL F. FAIRBANKS ◽  
LINDA T. LAMPE
Keyword(s):  
Dehydrogenase Activity ◽  
G6pd Deficiency ◽  
Dehydrogenase Deficiency ◽  
G6pd Activity ◽  
New Method ◽  
Cytochemical Method ◽  
Conventional Methods ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Individual Erythrocytes

Abstract A new method is described for the graded estimation of G6PD activity in individual erythrocytes. The method appears to offer some technical advantages over other methods. It requires reagents that are neither hazardous nor unstable and such equipment as might be found in the usual hematology laboratory. It appears to be more sensitive than conventional methods for identification of women heterozygous for G6PD deficiency and further substantiates the existence of erythrocyte mosaicism in such individuals.

G6PD Deficiency Prevalence as a Cause of Neonatal Jaundice in a Neonatal Ward in Dohuk, Iraq

2019 ◽  
Author(s):  
Adil Abozaid Eissa ◽  
Bijar Ali Haji ◽  
Adnan Anwar Al-Doski
Keyword(s):  
G6pd Deficiency ◽  
Complete Blood Count ◽  
Underlying Mechanism ◽  
Total Serum ◽  
Current Case ◽  
Indirect Bilirubin ◽  
Significant Difference ◽  
The Difference ◽  
Glucose 6 Phosphate Dehydrogenase

Abstract Objective The current study initiated to address the effect of glucose-6-phosphate dehydrogenase (G6PD) deficiency on the pathogenesis and the severity of neonatal hyperbilirubinemia (NHB). Study Design A total of 100 newborns with moderate to severe indirect hyperbilirubinemia and 50 normal neonates without hyperbilirubinemia had been enrolled in the current case–control study. All enrolled neonates had been tested for ABO and Rh(D) blood grouping, Total serum bilirubin measurement, complete blood count, morphology, reticulocyte counts, direct Coombs' test, and G6PD enzyme assay. Results From all enrolled hyperbilirubinemic neonates, 16% were G6PD deficient and this displays a statistically significant difference in comparison to controls (only 6% were G6PD deficient). Also, significant difference was found in the level of serum indirect bilirubin among G6PD-deficient neonate in comparison to G6PD nondeficient neonates which had contributed significantly to the difference in the duration of phototherapy and hospitalization among deficient neonate. Despite this, no significant difference found in the onset of presentation, reticulocytes count, and age of neonates between the two groups (G6PD-deficient and G6PD nondeficient neonates). Conclusion The current study augments the etiological role of G6PD in the causation and severity of NHB in the region; however, in the absence of significant difference in the reticulocytes and the hemoglobin level, the underlying mechanism cannot be backed to the excess hemolysis alone.

scholarly journals Comparison of Extracapsular Stabilization Techniques Using an Ultrasonically Implanted Absorbable Bone Anchor (Weldix) after Cranial Cruciate Ligament Rupture in Cats—An In Vitro Study

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1695
Author(s):  
Lydia Koch ◽  
Barbara Bockstahler ◽  
Alexander Tichy ◽  
Christian Peham ◽  
Eva Schnabl-Feichter
Keyword(s):  
Internal Rotation ◽  
Ex Vivo ◽  
Cruciate Ligament ◽  
Joint Stability ◽  
Bone Anchor ◽  
Ex Vivo Model ◽  
Cranial Cruciate Ligament ◽  
Stifle Joint ◽  
Significant Difference

Background: This study evaluated joint stability after surgical repair of cranial cruciate ligament (CrCL)-deficient stifle joints in cats using a novel absorbable polylactide bone anchor in an ex vivo model. Methods: Thirty-six hindlimbs from cats with intact (Gi group) and transected CrCLs were treated with fabellotibial suture alone (GFW group), suture combined with an absorbable polylactide bone anchor (GWD group), or suture combined with a nonabsorbable bone anchor (GFT group), positioned in a limb press with predefined joint angles (stifle joint: 120 ± 5°; hock joint: 120 ± 5°) and loaded with 10%, 20%, and 30% of body mass (BM). Predefined points were measured on lateral radiographs and with a coordinate measurement machine. Distances on radiographs (mm) were measured and angles (°) were calculated to represent the craniocaudal movement and the internal rotation of the tibia. Results: There were no differences for craniocaudal movement between Gi and GFW or GFT, but for GWD regarding angle measurement at 30% BM. For internal rotation, there was no significant difference between Gi and GFW or GWD, but for GFT. Conclusion: The used absorbable polylactide bone-anchor was able to stabilize the stifle joint regarding internal rotation and craniocaudal movement as calculated from distance measurements.

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