scholarly journals Establishment of a stable transgenic g6pdM1315-1443 zebrafish line with glucose-6-phosphate dehydrogenase deficiency

2020 ◽  
Author(s):  
Lu-Jun Shang ◽  
Jin Song ◽  
Hai-Xiong Xia ◽  
Yuan-Yuan Tuo ◽  
Ping-Ping Ren ◽  
...  
Keyword(s):  
G6pd Deficiency ◽  
Human Subjects ◽  
Clinical Manifestations ◽  
Transgenic Zebrafish ◽  
Hemoglobin Content ◽  
Zebrafish Model ◽  
Disease States ◽  
Clinical Syndromes ◽  
Pericardial Edema ◽  
Glucose 6 Phosphate Dehydrogenase

ABSTRACTGlucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common inherited enzymopathy in humans and is associated with a predisposition to hemolysis. However, there are few animal models to that adequately mimic associated human disease states that could be used to evaluate strategies to address clinical syndromes attributable to G6PD deficiency. In the present study, we aimed to establish a stable transgenic zebrafish model of G6PD deficiency that recapitulates the clinical manifestations of G6PD deficiency. We incorporated a stable transgene of G6PD lacking nucleotides from 1315 to 1443 denoted Tg(zgata1:g6pdM1315-1443-egfp). Functional analysis showed that Tg(zgata1:g6pdM1315-1443-egfp) transgenic zebrafish demonstrate a decrease in g6pd activity, reduced GSH levels and hemoglobin content, and increases in pericardial edema in response to α-naphthol exposure, similar to human subjects with G6PD deficiency. We detected no other significant phenotypic abnormalities compared to controls. Taken together, these observations indicate that the Tg(zgata1:g6pdM1315-1443-egfp) zebrafish line mirrors key clinical manifestations of G6PD deficiency in humans. This model may facilitate mechanistic studies and promote translational research related to G6PD deficiency.

scholarly journals Establishment of a stable zg6pdM118-144 transgenic zebrafish model of glucose-6-phosphate dehydrogenase deficiency

2020 ◽  
Author(s):  
Hai-Xiong Xia ◽  
Yan-Hua Zhou ◽  
Yuan-Yuan Tuo ◽  
Ping-Ping Ren ◽  
Jin Song ◽  
...  
Keyword(s):  
G6pd Deficiency ◽  
Genetic Defect ◽  
Clinical Manifestations ◽  
Wide Distribution ◽  
Health Concern ◽  
Transgenic Zebrafish ◽  
Zebrafish Model ◽  
Malaria Therapy ◽  
Glucose 6 Phosphate Dehydrogenase

AbstractGlucose-6-phosphate dehydrogenase (G6PD) deficiency, the most common genetic defect and enzymopathy with a wide distribution and increased public health concern, predisposes subjects succumb to oxidative stress. G6PD deficiency has been associated with hemolysis. Clinically, G6PD deficiency is asymptomatic and the clinical manifestations occur with the exposure to certain agents. Due to the lack of suitable animal models that can predict the clinical hemolytic potential of drugs, it needs an appropriate research model to fully recapitulate the manifestations of G6PD deficiency in clinic, to optimize the malaria therapy and promote anti-malarias development. The present study has displayed a stable transgenic Tg(zgata1-g6pdM118-144-egfp) zebrafish model with G6PD deficiency which mimics the clinical features of G6PD deficiency phenotypically and functionally. The findings showed that there was an inadequate level of reduced GSH in the transgenic Tg(zgata1-g6pdM118-144-egfp) zebrafish line in the presence or absence of α-naphthol, compared to the wildtype zebrafish, indicating an attenuation of g6pd activity in the transgenic zebrafish line. In addition, the observations show that there is a less abundance of g6pd in the transgenic Tg(zgata1-g6pdM118-144-egfp) zebrafish line. On the other hand, there is no morphological abnormality in the transgenic Tg(zgata1-g6pdM118-144-egfp) zebrafish line. Taken together, our work has delivered a novel stable transgenic zebrafish model of G6PD deficiency that will facilitate the mechanistic and functional elucidation for the role of G6PD in erythrocytic pathophysiology. This model will promote the translational research for the drug development, in particular, for anti-malarias development.

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.

Clinical Manifestations and Therapeutic Findings of the Children with Glucose-6-Phosphate Dehydrogenase Deficiency Presenting Favism

2020 ◽  
Vol 20 ◽  
Author(s):  
Fariba Tarhani ◽  
Alireza Nezami ◽  
Ghobad Heidari ◽  
Babak Abdolkarimi
Keyword(s):  
G6pd Deficiency ◽  
Dehydrogenase Deficiency ◽  
Clinical Manifestations ◽  
Descriptive Study ◽  
Cross Sectional ◽  
Common Complaint ◽  
The Mean ◽  
Fava Beans ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Hydration Therapy

Aim: Favism is characterized as acute anemia, due to Glucose-6-phosphate dehydrogenase (G6PD) deficiency as a result of fava beans intake. It is associated with paleness, jaundice, and hemoglobinuria. In this study, signs, symptoms and therapeutic findings of the patients with hemolysis due to G6PD deficiency were investigated in Shahid Madani Hospital of Khorramabad, Lorestan. Methods: This is a single-center cross-sectional descriptive study that was conducted on all children with G6PD deficiencyinduced hemolysis. Results: 308 children (64.3% male and 35.7% female) were included in this study. The most common complaint was jaundice (82.5%) and the most common cause of hemolysis was the intake of fava bean (85.7%). 68% of the children were treated with hydration/fluid therapy. Blood transfusion was conducted in 36.36% of the cases and the mean of blood administered was 18.9 cc/kg. Conclusion: In this study, hydration therapy was performed in most of the children presenting favism. Also, the incorrect calculation of the amount of blood needed for transfusion has increased the frequency of blood transfusions and prolonged hospitalization time.

scholarly journals Late-Life Presentation of Unsuspected G6PD Deficiency

2018 ◽  
Vol 2018 ◽  
pp. 1-3
Author(s):  
Marcos Benchimol ◽  
Laura Bernardo Madeira ◽  
Ricardo de Oliveira-Souza
Keyword(s):  
G6pd Deficiency ◽  
Clinical Manifestations ◽  
Diagnostic Challenge ◽  
Packed Red Blood Cells ◽  
Wide Range ◽  
Hemolytic Crisis ◽  
History Of ◽  
Triggering Conditions ◽  
Red Blood Cell Count ◽  
Glucose 6 Phosphate Dehydrogenase

Deficiency of glucose-6-phosphate dehydrogenase (G6PD) is the commonest enzyme deficiency in humans with a wide range of possible clinical manifestations depending on the specific genetic variant in each case. Here we present the case of an 86-year-old male of African descent who acutely developed symptoms of G6PD deficiency immediately after he received methylene blue for treating methemoglobinemia. The contrast between a low SO2 on pulse oximetry and a normal arterial gas sampling raised the possibility of methemoglobinemia. The patient was treated with packed red blood cells and folic acid, and a rapid clinical improvement followed by normalization of the red blood cell count ensued. In view of the patient’s advanced age, the lack of a history of similar episodes in the past, and the normal laboratory results during the hemolytic crisis, this case remained a diagnostic challenge for over three months, when a follow-up measure of G6DP activity eventually confirmed the diagnosis. A latent deficiency of G6PD may become clinically manifest under the appropriate triggering conditions even in elderly patients and in the absence of past or current clinical and laboratory evidence of G6PD deficiency.

Pro-Oxididant Induce Hemolysis in a Zebrafish Model of Glucose-6-Phosphate Dehydrogenase Deficiency.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2078-2078
Author(s):  
Xiaobai Patrinostro ◽  
Ashley C Kramer ◽  
Troy Lund
Keyword(s):  
Flow Cytometry ◽  
Free Radical ◽  
Mouse Models ◽  
G6pd Deficiency ◽  
Low Dose ◽  
Free Radical Production ◽  
Radical Production ◽  
Pericardial Edema ◽  
Oxidative Challenge ◽  
Glucose 6 Phosphate Dehydrogenase

Abstract Abstract 2078 Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common genetic defect and enzymopathy worldwide with an estimated 400 million persons affected. G6PD deficiency causes acute hemolysis in children exposed to pro-oxidants. Mouse models have suffered from their inaccuracy recapitulating the disease because of early death or lack of significant hemolysis in response to oxidative challenge. The zebrafish is an expanding model for many hematopoietic diseases. It's ex utero development and optical clarity offer advantages not obtained using mouse models. We targeted morpholinos to the 5-prime coding region of zebrafish g6pd. The g6pd morphants showed a significant reduction in G6pd protein expression and G6pd enzyme activity. We found a dose-dependant phenotype in zebrafish g6pd morphants. Low dose morpholino, 1.2 pmols, embryos displayed no overt phenotype, while high dose, 1.5 pmols, g6pd morphants displayed over a 50% reduction in levels of hemoglobin as determined by o-dianisidine staining. This hemolysis resulted in a phenotype displaying severe pericardial edema, which could be rescued by injection of zebrafish g6pd mRNA. Embryos injected with random morpholinos displayed no abnormalities. Common pro-oxidants include menthol, as a therapeutic ointment used in West Africa; naphthalene, an active ingredient in mothballs; and more commonly antimalaria drugs such as primaquine. We next tested whether an oxidative challenge on the low dose morpholino injected embryos by treatment with menthol or naphthol (the active form of naphthalene). We found these compounds significantly increased free radical production using a 6-carboxy-2′,7′-dichlorodihydrofluorescein diacetate fluorescence-based assay as well as an increase in hydroethidine staining, another indicator of free radical production. When low dose morphants were exposed to either compound, they displayed massive cardiac edema and significantly reduced hemoglobin levels due to hemolysis. Hemolysis was verified by performing similar experiments in fish with labeled erythrocytes, gata1:DsRed where we found a 60% reduction in gata1 positive cells by flow cytometry. Imagestream flow cytometry revealed that remaining erythrocytes stained positive for annexin V indicating active hemolysis. We were also able to show hemolysis and massive pericardial edema also occurred with exposure to the prototypical antimalaria drug, primaquine, which displayed increased reactive oxygen species at day 6 –7 post fertilization. In conclusion, we have faithfully developed a vertebrate model of G6PD deficiency in which in vivo hemolytic crisis occurs after exposure to pro-oxidants similar to what occurs in children. These compounds pose significant health risks to infants and children with G6PD deficiency. Utilizing this model, we will learn new information about novel cell processes and metabolic pathways that are disrupted in G6PD deficiency driven hemolysis. In addition, we will be able to screen potential anti-oxidants for their ability to reduce hemolysis. With the ultimate goal to attenuate the morbidity caused by the severe hemolytic crises occurring in children with G6PD deficiency. Disclosures: No relevant conflicts of interest to declare.

scholarly journals MicroRNA-21 Plays Multiple Oncometabolic Roles in the Process of NAFLD-Related Hepatocellular Carcinoma via PI3K/AKT, TGF-β, and STAT3 Signaling

Cancers ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 940
Author(s):  
Chi-Yu Lai ◽  
Kun-Yun Yeh ◽  
Chiu-Ya Lin ◽  
Yang-Wen Hsieh ◽  
Hsin-Hung Lai ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Signaling Pathways ◽  
Stellate Cell ◽  
Human Cancer ◽  
Tumor Formation ◽  
Transgenic Zebrafish ◽  
Zebrafish Model ◽  
Lipid Metabolism Disorder ◽  
Full Spectrum ◽  
Stat3 Signaling

MicroRNA-21 (miR-21) is one of the most frequently upregulated miRNAs in liver diseases such as nonalcoholic fatty liver disease (NAFLD) and hepatocellular carcinoma (HCC). However, mechanistic pathways that connect NAFLD and HCC remain elusive. We developed a doxycycline (Dox)-inducible transgenic zebrafish model (LmiR21) which exhibited an upregulation of miR-21 in the liver, which in turn induced the full spectrum of NAFLD, including steatosis, inflammation, fibrosis, and HCC, in the LmiR21 fish. Diethylnitrosamine (DEN) treatment led to accelerated liver tumor formation and exacerbated their aggressiveness. Moreover, prolonged miR-21 expression for up to ten months induced nonalcoholic steatohepatitis (NASH)-related HCC (NAHCC). Immunoblotting and immunostaining confirmed the presence of miR-21 regulatory proteins (i.e., PTEN, SMAD7, p-AKT, p-SMAD3, and p-STAT3) in human nonviral HCC tissues and LmiR21 models. Thus, we demonstrated that miR-21 can induce NAHCC via at least three mechanisms: First, the occurrence of hepatic steatosis increases with the decrease of ptenb, pparaa, and activation of the PI3K/AKT pathway; second, miR-21 induces hepatic inflammation (or NASH) through an increase in inflammatory gene expression via STAT3 signaling pathways, and induces liver fibrosis through hepatic stellate cell (HSC) activation and collagen deposition via TGF-β/Smad3/Smad7 signaling pathways; finally, oncogenic activation of Smad3/Stat3 signaling pathways induces HCC. Our LmiR21 models showed similar molecular pathology to the human cancer samples in terms of initiation of lipid metabolism disorder, inflammation, fibrosis and activation of the PI3K/AKT, TGF-β/SMADs and STAT3 (PTS) oncogenic signaling pathways. Our findings indicate that miR-21 plays critical roles in the mechanistic perspectives of NAHCC development via the PTS signaling networks.

scholarly journals Author Correction: Stimulation of hepatocarcinogenesis by activated cholangiocytes via Il17a/f1 pathway in kras transgenic zebrafish model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mohamed Helal ◽  
Chuan Yan ◽  
Zhiyuan Gong
Keyword(s):  
Transgenic Zebrafish ◽  
Zebrafish Model ◽  
Stimulation Of

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

scholarly journals Modulation of Innate Immune Toxicity by Silver Nanoparticle Exposure and the Preventive Effects of Pterostilbene

2021 ◽  
Vol 22 (5) ◽  
pp. 2536
Author(s):  
Rong-Jane Chen ◽  
Chiao-Ching Huang ◽  
Rosita Pranata ◽  
Yu-Hsuan Lee ◽  
Yu-Ying Chen ◽  
...  
Keyword(s):  
Silver Nanoparticles ◽  
Good Alternative ◽  
Innate Immune ◽  
Toxic Effects ◽  
Transgenic Zebrafish ◽  
Zebrafish Model ◽  
Cytokines And Chemokines ◽  
Living Organisms ◽  
And Function ◽  
Immune Toxicity

Silver nanoparticles pose a potential risk to ecosystems and living organisms due to their widespread use in various fields and subsequent gradual release into the environment. Only a few studies have investigated the effects of silver nanoparticles (AgNPs) toxicity on immunological functions. Furthermore, these toxic effects have not been fully explored. Recent studies have indicated that zebrafish are considered a good alternative model for testing toxicity and for evaluating immunological toxicity. Therefore, the purpose of this study was to investigate the toxicity effects of AgNPs on innate immunity using a zebrafish model and to investigate whether the natural compound pterostilbene (PTE) could provide protection against AgNPs-induced immunotoxicity. Wild type and neutrophil- and macrophage-transgenic zebrafish lines were used in the experiments. The results indicated that the exposure to AgNPs induced toxic effects including death, malformation and the innate immune toxicity of zebrafish. In addition, AgNPs affect the number and function of neutrophils and macrophages. The expression of immune-related cytokines and chemokines was also affected. Notably, the addition of PTE could activate immune cells and promote their accumulation in injured areas in zebrafish, thereby reducing the damage caused by AgNPs. In conclusion, AgNPs may induce innate immune toxicity and PTE could ameliorate this toxicity.

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