Vitisin B inhibits influenza A virus replication by multi-targeting neuraminidase and virus-induced oxidative stress

The development of drug-resistant influenza and new pathogenic virus strains underscores the need for antiviral therapeutics. Currently, neuraminidase (NA) inhibitors are commonly used antiviral drugs approved by the US Food and Drug Administration (FDA) for the prevention and treatment of influenza. Here, we show that vitisin B (VB) inhibits NA activity and suppresses H1N1 viral replication in MDCK and A549 cells. Reactive oxygen species (ROS), which frequently occur during viral infection, increase virus replication by activating the NF-κB signaling pathway, downmodulating glucose-6-phosphate dehydrogenase (G6PD) expression, and decreasing the expression of nuclear factor erythroid 2-related factor 2 (Nrf2) antioxidant response activity. VB decreased virus-induced ROS generation by increasing G6PD expression and Nrf2 activity, and inhibiting NF-κB translocation to the nucleus through IKK dephosphorylation. In addition, VB reduced body weight loss, increased survival, decreased viral replication and the inflammatory response in the lungs of influenza A virus (IAV)-infected mice. Taken together, our results indicate that VB is a promising therapeutic candidate against IAV infection, complements existing drug limitations targeting viral NA. It modulated the intracellular ROS by G6PD, Nrf2 antioxidant response pathway, and NF-κB signaling pathway. These results demonstrate the feasibility of a multi-targeting drug strategy, providing new approaches for drug discovery against IAV infection.

Oxidative stress markers as early predictors of diabetes complications in Type 2 diabetic patients

Objectives: Type 2 diabetes mellitus (T2DM) is a complex disease that affects many organs. Oxidative stress plays a key role in the pathogenesis of insulin resistance and ?-cell dysfunction. Thus, the present study aimed to use oxidative stress markers as early predictors for the progression of diabetic complications. Materials and Methods: The study sample included 400 individuals (300 T2DM and 100 non-diabetic controls) aged from 35 to 59 years randomly selected from the outpatient clinic of the National Institute for Diabetes and Endocrinology. T2DM patients were divided into subgroups: Subgroup (1) patients without any complications, Subgroup (2) patients with diabetic nephropathy(DN) and Subgroup (3) patients with cardiovascular disorders (CVD). Biochemical markers of fasting blood glucose, glycated haemoglobin (HbA1C), glucose-6-phosphate dehydrogenase (G6PD), lactate, arginase, heme oxygenase-1 (HO-1), haemoglobin (Hb), triglycerides (TG), cholesterol, low-density lipoprotein (LDL-C), high-density lipoprotein (HDL-C), urea, creatinine, malondialdehyde (MDA), reduced glutathione (GSH), catalase (CAT) and nitric oxide (NO) were performed. Results: DM patients showed significant increases in body mass index, systolic blood pressure, diastolic blood pressure, FBS, HbA1C, cholesterol, TG, LDL-C and glomerular filtration rate, while HDL-C decreased. Significant increases were observed in HO-1, MDA and NO, while G6PD/lactate, GSH and CAT decreased in DM patients. The DN and CVD patients exhibited a significant increase in HO-1, MDA and NO; while G6PD/lactate, GSH and CAT decreased compared with DM patients. Receiver operating characteristic analysis showed that the sensitivity and specificity of oxidative stress markers were 66.67–100%. Conclusion: Hexose monophosphate (HMP)/glycolysis pathways are shifted during DM near glycolysis rather than HMP pathway to produce energy where the amount of glucose enters the cells is low, causing oxidative stress. Oxidative stress markers could be used as early predictors of diabetes complications.

The clinical value of fluorescence quantitative assay and G6PD/6PGD ratio method in the diagnosis of glucose-6-phosphate dehydrogenase deficiency and gene mutation / 中华检验医学杂志

Objective:To investigate the diagnostic value of fluorescence quantitative method and G6PD/6PGD ratio method in glucose-6-phosphate dehydrogenase (G6PD) deficiency and the type of gene mutation.Methods:A total of 1 201 patients (711 males and 490 females) with suspected G6PD deficiency in Shanghai Children′s Hospital were collected from June 2018 to March 2021. Fluorescence quantification method, G6PD/6PGD ratio method and multicolor melting curve were used to detects enzyme activity, ratio and gene mutation type. Comparison of each index and evaluation of its diagnostic efficiency were performed.Results:Among 1 201 suspicious samples, 163 cases (135 males and 28 females) were finally diagnosed. 156 cases were diagnosed by fluorescence quantitative method with a detection rate of 95.71%, and 140 cases were diagnosed by G6PD/6PGD ratio method with a detection rate of 85.89%. enzymatic activity of G6PD and ratio of G6PD/6PGD in male were significantly lower than female, and the differences were statistically significant ( U=642.5, 734.5, P<0.001). 112 cases received G6PD gene mutation detection and 92 cases were diagnosed, 74 were hemizygous mutations, 1 were homozygous mutations, 15 were heterozygous mutations, and 2 were compound heterozygous mutations. Among 15 cases of heterozygous mutations, 11 cases were diagnosed by fluorescence quantitative method, the diagnosed rate was 73.33%, 4 cases were diagnosed by G6PD/6PGD ratio method, and the diagnosed rate was 26.67%. A total of 7 mutation sites were detected and the proportions were c.1388G>A (32.22%), c.1376G>T (30.00%), c.871G>A (13.33%), c.1024C>T (11.11%). c.95A>G (7.78%), c.487G>A (4.44%), c.392G>T (1.11%). The enzymatic activities of c.1376G>T and c.1024C>T, c.487G>A were statistically significant ( P<0.001,0.015); the G6PD/6PGD ratios of c.1024C>T and c.1388G>A, c.1376G>T were statistically significant ( P=0.017,0.002,0.011,0.013). Fluorescence quantitative method had sensitivity of 100%, specificity of 95.65%, and the area under the curve (AUC) is 0.972. The sensitivity of the G6PD/6PGD ratio method was 100%, the specificity was 94.57%, and the AUC was 0.979. The sensitivity of fluorescence quantitative method combined with G6PD/6PGD ratio was 96.7%, the specificity was 100%, and the AUC was 0.992. Conclusions:Compared with fluorescence quantification, the G6PD/6PGD ratio method might not be able to diagnose female heterozygotes effectively; The panel of G6PD fluorescence quantification and G6PD/6PGD ratio was helpful to reduce the missed diagnosis. Combined with gene mutation analysis, it could improve the diagnosis rate of G6PD deficiency in the children.

G6PD Regulates Cyclin E1 and CDK2 to Promote Cell Proliferation and Its Prognostic Value in Clear Cell Renal Cell Carcinoma / 中国生物化学与分子生物学报

Clear cell renal cell carcinoma (ccRCC) has been proved to be a metabolic disease with high

Effect of weekly versus daily primaquine on Plasmodium vivax malaria recurrences: A real-life cohort study

ABSTRACT Background: Although primaquine (PQ) is indicated for G6PD-deficient patients, data on weekly PQ use in Brazil are limited. Methods: We aimed to investigate malaria recurrences among participants receiving daily and weekly PQ treatments in a real-life setting of two municipalities in the Amazon between 2019 and 2020. Results: Patients receiving weekly PQ treatment had a lower risk of recurrence than those receiving daily PQ treatment (risk ratio: 0.62, 95% confidence interval: 0.41-0.94), using a model adjusted for study site. Conclusions: Weekly PQ use did not increase the risk of malaria recurrence. Further studies with larger populations are warranted.

Research progress of glucose 6 phosphorus dehydrogenase in malignant tumor / 中国临床药理学与治疗学

Glucose-6-phosphate dehydrogenase (Glucose-6-phosphate dehydrogenase, G6PD) is the rate-limiting enzyme of pentose phosphate pathway (PPP), which mainly maintains the balance of NADPH and intracellular redox reaction. Reducing G6PD activity or PPP dysfunction can prevent normal cell proliferation, and severe lack of G6PD can damage embryonic development and delay organ growth. At present, many studies have proved that abnormal activation of G6PD can lead to the enhancement of cell proliferation and adaptability of various types of cancer, and it is easy to cause drug resistance and increase the difficulty of clinical treatment. It has become an urgent need for clinical treatment to study the mechanism of G6PD in cancer cells and identify new potential drug therapeutic targets.

Prevalence of UGT1A1 (TA)n promoter polymorphism in Panamanians neonates with G6PD deficiency

A relationship between the polymorphism in promoter region of the UGT1A1 gene and the development of jaundice has been demonstrated recently. This polymorphism leads to 30% of normal rate transcription initiation of UGT1A1 gene, thus decreasing the bilirubin glucuronidation. The combination of the G6PD deficiency and polymorphism in neonates and adults may cause pronounced hyperbilirubinaemias. The aim of this study was to analyse the variations in the UGT1A1 gene promoter in Panamanians neonates with G6PD deficiency and its association with neonatal jaundice (NJ). We identified five different genotypes of TA repeats, in 17 neonates (42.5%) the normal variant TA6/TA6 and in the other 57.5% of the subjects: TA7/TA7 (12.5%), TA6/TA7 (40%), TA6/ TA8 (2.5%) and TA6/TA5 (2.5%). Additionally 75% of the 16 newborns that showed NJ had an abnormal variant in the promoter sequence, although, there was no significant difference (P = 0.068). The risk of jaundice in neonates with TA7 variant was thrice higher in subjects than with other alleles (P = 0.093, CI: 0.81–11.67). The TA7 allele frequency in this study (0.325) was consistent with the global frequency and similar to Caucasians. The results proved that there is no significant relationship between promoter polymorphism in UGT1A1 and NJ in G6PD deficient Panamanian newborns. Further studies with a greater number of subjects would determine the exact relationship between marked NJ and UGT1A promoter variations.

Effect of Qingzao Jiufei Tang on Enzymatic Activity and Regulatory Factor of Key Enzyme G6PD in Pentose Phosphate Energy Metabolism Pathway in Lung Cancer / 中国实验方剂学杂志

Objective::To discuss the effect of Qingzao Jiufei Tang on enzymatic activity and regulatory factor of glucose 6-phosphatedehydrogenase(G6PD) in pentose phosphate energy metabolism pathway in lung cancer. Method::Fifty male C57BL6J mice were randomly divided into five groups. Animal models were induced through axillary injection with Lewis cells. The Qingzao Jiufei Tang group was given drugs (11, 5.5, 2.8 g·kg-1·d-1) two weeks before modeling, the cyclophosphamide(CTX) group was intraperitoneally injected with CTX (50 mg·kg-1), and the model group was intraperitoneally injected with the same volume of saline after molding. At 14 d after modeling, the mice were sacrificed, and the tumor tissues were collected. The enzymatic activity of G6PD, content of reactive oxygen species (ROS) were detected by enzyme-linked immunosorbent assay(ELISA) method. Expressions of gp91phox and p22phox mRNA were detected by real-time fluorescent quantitative polymerase chain reaction(Real-time PCR) method. Result::Compared with the model group, the enzymatic activity of G6PD in high-dose group, medium-dose group and low-dose group were reduced obviously (P<0.05, P<0.01). Content of ROS, mRNA expressions of gp91phox and p22phox in high-dose group, medium-dose group and low-dose group were reduced obviously (P<0.01). Conclusion::Qingzao Jiufei Tang may inhibit the expression of G6PD by inhibiting the expression of gp91 phox, p22phox oxidase, and then reduce content of ROS, so as to reduce the energy metabolism and hyperplasia of lung cancer cells.

Prevalence, Sociodemographic and Clinical Characteristics of G6PD Deficient Blood Donors in Terengganu and the Effects of Storage on Their Donated Blood

@#Introduction: Glucose-6-phosphate dehydrogenase (G6PD) deficiency causes red blood cell destruction due to oxidative stress. G6PD is essential for NADPH conversion; which is critical for glutathione reductase to prevent damage to cellular structures. In Malaysia, blood donors are not routinely screened for G6PD deficiency. We hypothesise that G6PD-deficient red blood cells are more likely to haemolyse during storage due to increased oxidative molecules. The objectives of this study were to determine the prevalence of G6PD deficiency among blood donors, describe their characteristics and to evaluate the effects of storage on G6PD-deficient donated blood. Methods: This study was conducted at selected mobile donation centres in Terengganu. Consented blood donors were screened for G6PD status using fluorescent spot tests (FST). G6PD enzyme activities were measured for donors who were G6PD deficient. Effects of storage on haemolysis from G6PD-deficient donors were compared with non G6PD-deficient group. Sixty ml of blood was collected from blood unit to transfer pouch for estimation of haemoglobin (Hb), plasma Hb, percentage of haemolysis and plasma potassium. Serial sampling with a 7-day interval was done from Day 1 to Day 35. Statistical analysis was considered significant if p ≤0.05. Results: A total of 440 blood donors were screened and 12 male donors were found to be G6PD deficient by FST. Enzymatic activities were measured in 11 donors as one donor sample failed to be sent to the centre due to logistic problem. Their enzymatic activities ranged from 1.66-2.93 U/g Hb whereby 6 have severe deficiency and the other 5 were categorised as partial deficiency. Donors were asymptomatic for haemolytic episode. Serial sampling showed there was no significant difference of haemolytic parameters in blood units of G6PD-deficient donors as compared to control (p>0.05). Conclusion: Prevalence of G6PD blood donors in Terengganu mobile centres was 2.7%. G6PD enzyme activities did not correlate with clinical symptoms. Haemolytic parameters were not affected in blood units which were G6PD-deficient.

Recent trends of glucose-6-phosphate dehydrogenase deficiency among saudi population in Riyadh city / Tendências recentes da deficiência de glicose-6-fosfato desidrogenase entre a população saudita na cidade de Riade

G6PD deficiency is associated with erythrocyte deficiency in the X-chromosome enzyme. It causes a hematologic syndrome called hemolytic anemia that connects G6PD deficiency with X-linked condition. In the Middle East, including Saudi Arabia, G6PD deficiency is the most dominant genetic blood disorders. It results in higher rates of mortality and morbidity due to its incurable long-lasting nature and prevalence of physical and psychological incapacities. In this study, an attempt was made to evaluate the prevalence of G6PD deficiency among the Saudi population in Riyadh city. A cross-sectional retrospective study was conducted at King Saud University Medical City in Riyadh, Saudi Arabia. The population of the study comprised randomly chosen males and females who visited the hospital from January 2017 to January 2018. Statistical analyses were performed using SPSS, and descriptive analysis was used to find the frequency of G6PD-deficient patients. Out of the 209 patients, 62.2% were males (n=130) and 37.8% were females (n=79). Twenty males and 6 females were found to have G6PD deficiency, with the male to female ratio being 1:3. Out of the total 130 male participants, 20 patients were found to be enzyme deficient and 6 patients of 79 female patients were found to be G6PD deficient. There were 38.4% (n=10) patients with G6PD level <4 units/gram hemoglobin, 26.9% (n=7) patients had G6PD levels of 4.1­7.0 units/gram hemoglobin, and 34.6% (n=9) patients had >7 units/gram hemoglobin. Among the G6PD patients, 23.07% patients were severely anemic, and 5 (19.2%) patients were reported to have high bilirubin. The present study revealed the G6PD prevalence to be 12.4% among the Saudi population; this value is significantly higher than that found in France, Spain, India, and Singapore. In the Saudi population, males are more vulnerable to G6PD-deficient than females. Hence, attention should be paid to G6PD-deficient patients while prescribing antimalarial medication. Such patients may be advised to avoid certain foods to minimize the risk of having hemolytic episodes.

A deficiência de G6PD está associada à deficiência de eritrócitos na enzima do cromossomo X. Causa uma síndrome hematológica chamada anemia hemolítica que conecta a deficiência de G6PD à condição ligada ao X. No Oriente Médio, incluindo a Arábia Saudita, a deficiência de G6PD é o distúrbio genético do sangue mais dominante. Isso resulta em maiores taxas de mortalidade e morbidade devido à sua natureza incurável e duradoura e à prevalência de incapacidades físicas e psicológicas. Neste estudo, foi feita uma tentativa de avaliar a prevalência de deficiência de G6PD entre a população saudita na cidade de Riade. Um estudo retrospectivo transversal foi realizado na cidade médica da Universidade King Saud, em Riade, na Arábia Saudita. A população do estudo compreendeu homens e mulheres escolhidos aleatoriamente que visitaram o hospital entre janeiro de 2017 e janeiro de 2018. As análises estatísticas foram realizadas com o SPSS e a análise descritiva foi utilizada para determinar a frequência de pacientes com deficiência de G6PD. Dos 209 pacientes, 62,2% eram do sexo masculino (n = 130) e 37,8% eram do sexo feminino (n = 79). Verificou-se que 20 homens e 6 mulheres apresentavam deficiência de G6PD, sendo a proporção homem/mulher de 1:3. Do total de 130 participantes do sexo masculino, 20 pacientes apresentaram deficiência de enzima e 6 de 79 pacientes do sexo feminino apresentaram deficiência de G6PD. Havia 38,4% (n = 10) pacientes com nível de G6PD < 4 unidades/grama de hemoglobina, 26,9% (n = 7) pacientes tinham níveis de G6PD de 4,1-7,0 unidades/grama de hemoglobina e 34,6% (n = 9) pacientes tinham > 7 unidades/grama de hemoglobina. Entre os pacientes com G6PD, 23,07% eram severamente anêmicos e cinco (19,2%) pacientes relataram ter alta bilirrubina. O presente estudo revelou que a prevalência de G6PD é de 12,4% na população saudita; esse valor é significativamente maior que o encontrado na França, Espanha, Índia e Cingapura. Na população saudita, os homens são mais vulneráveis à deficiência de G6PD do que as mulheres. Portanto, deve-se prestar atenção aos pacientes com deficiência de G6PD durante a prescrição de medicamentos antimaláricos. Esses pacientes podem ser aconselhados a evitar certos alimentos para minimizar o risco de episódios hemolíticos.

Abnormalities of Hemoglobin and Glucose-6-Phosphate-Dehydrogenase Deficiency in Children with Uncomplicated Malaria and Living in Banfora and Saponé, Two Different Malaria Setting of Burkina Faso

Aims:The aim of this study is to assess the prevalence of hemoglobin abnormalities and G6PD deficiency and their respective influence on anemia occurring in less than five years old children with clinical P. falciparummalaria living in Burkina Faso.Study Design:The study was a cross-sectional survey with descriptive focus conducted from December 2010 to January 2013 in Saponé health district and from May to October 2011 in Banforahealth district. Clinical and laboratory data were collected. Blood smears on slides for malaria diagnosis by microscopy, hemoglobin level and filter paper for the detection of human genetic factors were performed.Methodology:A total of 386 subjects from Saponé (131) and Banfora (255) were enrolled. DNA collected from each sample was extracted using chelex-100 method and the human genetic resistance factors background was assessed by RFLP-PCR. Abnormal hemoglobin patients were classified as NonAA while AA was defined the normal hemoglobin.Results:In this study, 70.98% (274/386) were classified normal hemoglobin (AA) while 29.02% (112/386) of subjects were carrying at least one abnormal (NonAA) allele: 24.35%AC, 3.63% AS, 0.78%CC and 0.26%SC. G6PD deficiency was 9.59% (37/386) among which, 4.92% for male and 4.66% in female. However, this gender difference was not statistically significant (p=1.00). 319/367 (86.92%) of the patients were anemic (59.4% with moderate anemia and 20.98% with mild anemia). Theprevalence of anemia in G6PD deficient subjects was 83.33% (of which 58.33% were moderate anemia and 22.22% mild anemia). The difference between types of hemoglobin (p=0.64) in the occurrence of anemia (AA 87.64% and Non AA 85.18%) was not statistically significant. Conclusion:This study showed that the prevalence of these genetic factors was relatively low among children with clinical falciparummalaria with high parasite density. In addition, these factors appear to have no effect on anemia.

The Impact of Human Genetic Factors (G6pd and Type of Hemoglobin) on the Course of Uncomplicated Malaria Infection in Children Aged from 2 to 10 Years Living in the Banfora Health District in Burkina Faso

Aims: The aim of this study was to assess the impact of hemoglobin polymorphisms and G6PD deficiency on the course of uncomplicated malaria infection in children aged from 2 to 10 years in Burkina Faso.Study Design: The study was conducted as a longitudinal study in Banfora health district. A total of 150 children aged from 2 to 10 years was enrolled and followed up between May 2015 and February 2016. Blood samples were collected at four different time points: before infection (Visit 1), during asymptomatic parasitemia (Visit 2), during symptomatic parasitemia (Visit 3) and three weeks after treatment (Visit 4). Clinical examination, hematology parameters and malaria diagnosis using microscopy were performed. Hemoglobin and G6PD typing were done using PCR-RFLP. Hemoglobin AA genotypes were defined as normal hemoglobin while Hemoglobin AC, AS and SS were defined as abnormal hemoglobin (hb non-AA).Results:The prevalence of hemoglobin (hb) genotypes was 81.21% for AA while hb non-AA genotypes were estimated at 18.79% (12.08% for hbAC, 6.04% for hbAS and 0.67% for HbSC). The prevalence of G6PD genotypes was 89.26% and 10.74% for normal G6PDn and G6PD deficiency respectively. The prevalence of asymptomatic carriers of P. falciparumwas not affected neither by the genotypes of Hemoglobin, nor by the G6PD deficiency. Conversely, the risks of developing uncomplicated malaria in G6PD deficiency (G202A) group, was significantly lower (p=0.04).The results showeda significant difference (p˂0.0001) in the means of P. falciparumparasite densities between asymptomatic and symptomatic phase in Hemoglobin AA genotypes carriers while the means of parasite density was comparable in non-Hemoglobin AA carriers. Conclusion:Our study showed that G6PD deficiency protects against clinical malaria while P. falciparumparasite density increasing was correlated with carrying hemoglobin genotypes AA.

Case Report on Methylene Blue Induced Hemolytic Anemia

A 22-years old male patient was admitted to a tertiary care hospital with complaints of an alleged history of intentional poisoning (organophosphorus compound and nitrofurantoin) and developed hemolytic anemia after receiving methylene blue for 8 days. The patient presented with hematuria and hemoglobin level 3.1 which confirmed hemolytic anemia G6PD level was normal. Methylene blue was discontinued and PRBC transfusion (3 pints) was given. After 4 days of blood transfusion, the patient’s Hb level became 9.4 g/dl. Causality assessment was suggestive of a probable relationship between the drug and reaction.

Assessment of the prevalence of G6PD deficiency in RBCs of live new-borns born at tertiary care hospital

Background: Glucose-6-phosphate dehydrogenase is one of many enzymes that help the body process carbohydrates and turn them into energy. The mechanism by which G6PD deficiency causes neonatal hyper bilirubin may be due to hemolysis, but other mechanisms like secondary impairment of bilirubin conjugation and clearance by the liver may play a role. Therefore, through this study authors attempt to study the need for a newborn screening program for G6PD deficiency because of high prevalence and high risk of incidence due to consanguineous marriages in India.Methods: This study was a prospective cross-sectional study conducted among 350 consecutively born live new-borns in maternity wards and NICU of Krishna Institute of Medical Sciences and Hospital and Research Centre, Karad, Maharashtra during October 2016 to October 2017.Results: The maximum numbers of newborns were in the age group of 0-10 hours (36.80%), followed by in 11-20 hours (21.80%). The mean age among newborns was 2.86'5.83 hours. Out of 350 cases females were 181 (51.71%) and males (48.29%) and female to male ratio was 1.07:1.Conclusions: G6PD deficiency is one of the major causes of neonatal jaundice within 24 hours of life in new-borns. Hence, neonatal screening for G6PD deficiency could be an alternative to the haemolytic crisis prevention strategy in order to optimize affected young child care and prevention of crisis occurrence by avoiding taking contraindicated foods and drugs.

Karyotyping and population genetics in Cold War Mexico: Armendares's and Lisker's characterization of child and indigenous populations, 1960s-1980s / Cariotipia e genética de populações no México da Guerra Fria: as caracterizações de Armendares e Lisker sobre as populações infantil e indígena, 1960-1980

Abstract This paper focuses on geneticists Salvador Armendares's and Rubén Lisker's studies from the 1960s to the 1980s, to explore how their work fits into the post-1945 human biological studies, and also how the populations they studied, child and indigenous, can be considered laboratories of knowledge production. This paper describes how populations were considered for different purposes: scientific inquiry, standardization of medical practices, and production or application of medicines. Through the narrative of the different trajectories and collaborations between Armendares and Lisker, this paper also attempts to show the contact of their scientific practices, which brought cytogenetics and population genetics together at the local and global levels from a transnational perspective.

Resumo Aborda o trabalho dos geneticistas Salvador Armendares e Rubén Lisker, entre 1960 e 1980, para analisar como se insere nos estudos biológicos humanos do pós-1945, e demonstra como as populações estudadas por eles, a infantil e a indígena, podem ser consideradas laboratórios de produção de conhecimento. O artigo revela como as populações foram consideradas para diversos propósitos: investigação científica, padronização das práticas médicas e produção ou aplicação de suas medicinas. Por meio da narrativa das diferentes trajetórias e colaborações entre Armendares e Lisker, também procura discutir o contato de suas práticas científicas, que colocaram a citogenética e a genética de populações lado a lado nos níveis local e global a partir de uma perspectiva transnacional.

Establishment of a stable HEK293T cell line with c.392G>T (p.131G>V) mutation site knockout in gene using CRISPR/Cas9 technique / 南方医科大学学报

OBJECTIVE@#To establish a stable HEK293T cell line with c.392G>T (p.131G>V) mutation site knockout in gene using CRISPR/Cas9 technique.@*METHODS@#We designed 4 pairs of small guide RNA (sgRNA) for c.392G>T(p.131G>V) mutation site, and constructed exogenous PX458 plasmids expressing Cas9-sgRNA. The plasmids were transfected into HEK293T cells, and the cells expressing GFP fluorescent protein were separated by flow cytometry for further culture. After verification of the knockout efficiency using T7 endonuclease Ⅰ, the monoclonal cells were screened by limiting dilution and DNA sequencing to confirm the knockout. We detected the expressions of mRNA and protein and examined functional changes of the genetically modified cells.@*RESULTS@#We successfully constructed the Cas9-sgRNA exogenous PX458 plasmid based on the c.392G>T(p.131G>V) mutation site of gene. The editing efficiency of the 4 pairs of sgRNA, as detected by T7E1 enzyme digestion, was 6.74%, 12.36%, 12.54% and 2.94%. Sanger sequencing confirmed that the HEK293T cell line with stable knockout of c.392G>T(p.131G>V) was successfully constructed. The genetically modified cells expressed lower levels of mRNA and protein and showed reduced enzyme activity and proliferative capacity and increased apoptosis in response to vitamin K3 treatment.@*CONCLUSIONS@#We successfully constructed a stable HEK293T cell model with gene c.392G>T(p.131G>V) mutation site knockout to facilitate future study of gene repair.

Genetic analysis of glucose-6-phosphate dehydrogenase deficiency among newborns in Zhejiang Province / 中国实用儿科杂志

OBJECTIVE: To analyze the characteristics of gene mutations of glucose-6-phosphate dehydrogenase(G6PD)deficiency in the neonates of Zhejiang Province,and discuss the genetic diversity. METHODS: A total of 2242G6PD values and blood stains on dry filter were collected from the children borned in Zhejiang Province with positive screening of G6PD deficiency between March 2015 and September 2017 in Neonatal Metabolic Screening Center of Zhejiang Province. Genomic DNA of the stains were extracted. Thirty-five gene mutation sites were detected by Mass ARRAY technology. The relationship between mutation gene sites and G6PD activity were analyzed by SPSS 22.0,taking PT,c.1388G>A,c.1024C>T,c.95 A>G,c.871G>A and c.392G>T accounted for 92.96%. There were statistically significant differences in G6PD activity of c.1376 G>T,c.1388G>A,c.1024 C>T,c.95A>G(PT has obvious regional characteristics. Using MassARRAY technology to detect specific mutation sites can be a choice as a second-level screening method for G6PD deficiency.

Prevalence of Glucose-6-Phosphate Dehydrogenase Deficiency among Neonates in Usmanu Danfodiyo University Teaching Hospital (UDUTH), Sokoto, Nigeria: Oxidative Stress Markers in G6pd Deficient Neonates

Background: Glucose-6-phosphate dehydrogenase deficiency is one of the most common enzyme defects affecting all races and particularly in malaria-endemic areas. This study aimed at determining G6PD deficiency, bilirubin and oxidative stress biomarkers in G6PD deficient neonates among neonates in UDUTH, Sokoto. Methods: Samples of cord blood were collected at delivery, in the Labour Room, from 300 neonates made up of 131 (43.7%) males and 169 (56.3%) females. Methaemoglobin reduction method was used for the screening of G6PD deficiency; total bilirubin level was estimated using bilirubinometer, total antioxidant capacity (TAC) was measured using TAC Assay Kit, and malondialdehyde (MDA) using thiobarbituric acid method. Results: Of the 300 neonates tested, a total of 90(30%) were G6PD-deficient while 210(70%) had normal G6PD status. Of the 90 G6PD-deficient neonates, 41(45.6%) were males and 49(54.4%) were females. The prevalence was 31.3% among male population and 29.0% among female population. The mean ± standard error of total bilirubin (mg/dL), TAC (uM CRE), and MDA (nmol/L) in G6PD-deficient and G6PD-normal neonates were 6.63 ± 0.12 and 6.11 ± 0.06, 364.34 + 18.76 and 390.99 + 24.18, 26.15 + 1.22 and 23.35 + 1.15 respectively. The total bilirubin was significantly higher (p<0.05) in G6PD-deficient neonate than in G6PD-normal neonates, both TAC and MDA values showed no significant difference between the G6PD deficient and G6PD normal neonates. Conclusion: From this study, there is a high prevalence of G6PD deficiency among neonates in UDUTH, Sokoto. G6PD deficiency is a known cause of neonatal jaundice hence it is recommended G6PD screening be made routine for all neonates born in UDUTH, Sokoto.

Effect of knockdown or overexpression of G6PD on the proliferation and migration of hepatoma cells / 实用医学杂志

Objective To investigate the effect of knockdown or overexpression of G6PD on proliferation, growth and migration of human hepatocellular carcinoma cell PLC/PRF/5. Methods Lentivirus-mediated knock-down or overexpression of G6PD was achieved in human hepatocellular carcinoma cell line PLC/PRF/5. RT-PCR and Western blotting assay were used to detect the overexpression or knockdown of G6PD.Cell proliferation and mi-gration curves were recorded by real-time cell analysis system(RTCA),the cell proportion in the DNA replication phase can be directly displayed with EDU experiment,cell growth ability was detected by colony forming assay. Results The doubling time of cells in G6PD knockdown group was longer than that of the control group,and the cell growth rate decreased significantly,the proportion of cells in proliferative phase(43.2%)was lower than that in the control group,but the rates colony formation and migration were significantly decreased(P<0.05,respective-ly),and the migration curves separated apparently.While no significant differences in proliferation,growth and mi-gration of PLC/PRF/5 cells were found between the over-expressed strain and the control group. Conclusion The reduction of G6PD expression in HCC cells inhibits the proliferation and growth of HCC,which may lay a foun-dation for the further study of the pathogenesis and treatment of HCC.

Validation of the rapid test carestart(tm) G6PD among malaria vivax-infected subjects in the Brazilian Amazon

Abstract: INTRODUCTION: In the Brazilian Amazon, malaria infections are primarily caused by Plasmodium vivax. The only drug that kills the hypnozoite form of P. vivax is primaquine, thereby preventing relapse. However, treating glucose-6-phosphate dehydrogenase (G6PD)-deficient individuals with primaquine can lead to severe hemolysis. G6PD deficiency (G6PDd) affects approximately 400 million people worldwide, most of whom live in malaria-endemic areas. Therefore, clinicians need tools that can easily and reliably identify individuals with G6PDd. This study estimated the accuracy of the Carestart(tm) G6PD rapid test (Access Bio) in the diagnosis of G6PDd in male participants with and without P. vivax acute malaria. METHODS: Male participants were recruited in Manaus. Malaria diagnosis was determined by thick blood smear. G6PD quantitative analysis was performed spectro photometrically at a wave length of 340nm. The Carestart(tm) G6PD test was performed using venous blood. Genotyping was performed for individuals whose samples had an enzyme activity less than 70% of the normal value. RESULTS: Six hundred and seventy-four male participants were included in this study, of whom 320 had a diagnosis of P. vivax malaria. In individuals with enzyme activity lower than 30% (n=13), the sensitivity, specificity, positive predictive value, and negative predictive value of the Carestart(tm) G6PD test were as follows: 61.5% (95%CI: 35.5%-82.3%), 98.3% (95%CI: 97.0%-99.1%), 42.1% (95%CI: 23.1%-63.7%), and 99.2% (95%CI: 98.2%-82.3%), 98.3% (95%CI: 97.0%-99.1%), 42.1% (95%CI: 23.1%-63.7%), and 99.2% (95%CI: 98.2%-99.7%), respectively. Increases in sensitivity were observed when increasing the cut-off value. CONCLUSIONS: Despite low sensitivity, Carestart(tm) G6PD remains a good alternative for rapid diagnosis of G6PDd in malaria-endemic regions.