Aflatoxins are distributed worldwide throughout environment. As many as 30genes are involved in aflatoxin biosynthesis. aflD (nor-1) was predicted to encode a 31 kDa ketoreductase (AFLD), which catalyzed the earliest ...Aflatoxins are distributed worldwide throughout environment. As many as 30genes are involved in aflatoxin biosynthesis. aflD (nor-1) was predicted to encode a 31 kDa ketoreductase (AFLD), which catalyzed the earliest and first stable aflatoxin precursor in aflatoxin biosynthetic pathway. AFLD was proved to be necessary for conversion of norsolorinic acid (NA) to averatin (AVN) in aflatoxin biosynthesis. For this reason, aflD was cloned and specific monoclonal antibodies for AFLD were developed to better define potential pathways of AFLD involved in Aflatoxin B_1 (AFB_1) biosynthesis. Monoclonal antibodies 2A8-1E10 and 2F11-3D8 were successfully screened out by immunized mouse. Immunoblot analysis revealed that 2A8-1E10 had high sensitivity and specificity to identify native AFLD protein in A. flavus with detection limit of 3 ng/mL. These results showed that it was suitable for quantitative detection of AFLD in A. flavus isolate. Further investigation revealed that aflatoxin accumulations of various A. flavus were not dependent on AFLD biosynthesis.Overall, this is the first report for AFLD monoclonal antibody development and application in A. flavus quantitative detection.展开更多
The aflatoxin B1(AFB1)was considered as the most toxic natural compound and exists widely in the world,it is great significance to develop a high-performance sensor for simultaneously detection AFB1 and its toxin-prod...The aflatoxin B1(AFB1)was considered as the most toxic natural compound and exists widely in the world,it is great significance to develop a high-performance sensor for simultaneously detection AFB1 and its toxin-producing gene aflD.In this paper,a magnetically controlled fluorescent sensor was developed,ssDNA1/ssDNA2 was modified on graphene quantum dots(GQDs)/Mn:ZnS quantum dots(QDs)as an energy donor,and cDNA/Apt modified by black hole quencher(BHQ)was coupled with MNPs as an energy acceptor,which leaded to fluorescence quenching through donor-acceptor energy transfer.Parts of fluorescent probes were separated from magnetic carriers when the sensor was incubated with the targets.After magnetic separation,the fluo-rescence intensity of the fluorescent probe released from the supernatant was positively correlated with the target concentration.The simultaneous detection of aflD and AFB1 was realized.The sensor displayed a wide detection range(0.1-500 nM and 0.1-500 ng/mL)for aflD and AFB1,with detection limits of 0.021 nM and 0.038 ng/mL for aflD and AFB1.The magnetic fluorescence biosensor showed a good recovery rate in real rice flour and corn flour samples.The magnetic control fluorescence biosensor had the high sensitivity and speci-ficity.This strategy provided a simple,efficient and sensitive method for food safety monitoring.展开更多
Background and Aims:Metabolic dysfunction and obe-sity commonly coexist with both alcoholic and nonalcoholic fatty liver disease(AFLD and NAFLD).The association of AFLD and NAFLD with incident diseases in individuals ...Background and Aims:Metabolic dysfunction and obe-sity commonly coexist with both alcoholic and nonalcoholic fatty liver disease(AFLD and NAFLD).The association of AFLD and NAFLD with incident diseases in individuals with different metabolic phenotypes are unclear.Methods:UK Biobank study participants were screened for the presence of fatty liver at baseline.Body mass index and metabolic dysfunction were used to define metabolic phenotypes.Cox regression model was performed to examine the associations of AFLD and NAFLD with incident significant liver diseases(SLDs),cardiovascular diseases(CVDs),chronic kidney dis-eases(CKDs),and cancers,respectively.Results:A total of 43,974 AFLD and 103,248 NAFLD cases were identified.Both AFLD and NAFLD were associated with an increased risk of diseases of interest.The effects were amplified by obesity and metabolic abnormalities and modified by metabolic phe-notypes.Compared to individuals free of fatty liver and with phenotype of metabolically healthy-normal weight,AFLD[hazard ratio(HR 3.27;95%CI:1.95-5.47)]and NAFLD(HR 2.25;95%CI:1.28-3.94)cases with phenotype of met-abolically obese-normal weight had the greatest risk of SLDs.For CVDs,CKDs,and cancer,the greatest risks were detected in AFLD and NAFLD cases with phenotype of metabolically obese-overweight/obesity.In this subpopulation,AFLD and NAFLD conferred a 2.75-fold(95%CI:2.32-3.25)and 4.02-fold 95%CI:(3.64-4.43)increased risk of CVDs,4.37-fold 95%CI:(3.38-5.64)and 6.55-fold 95%CI:(5.73-7.48)increased risk of CKDs,and 1.19-fold 95%CI:(1.08-1.27)and 1.21-fold 95%CI:(1.14-1.28)increased risk of cancers,respectively.Conclusions:Metabolic phenotypes modified the association of AFLD and NAFLD with intrahepatic and ex-trahepatic diseases.展开更多
基金supported by National Natural Science Foundation of China (31471650, 31640062)National Key Research and Development Program of China (2016YFE0119900)
文摘Aflatoxins are distributed worldwide throughout environment. As many as 30genes are involved in aflatoxin biosynthesis. aflD (nor-1) was predicted to encode a 31 kDa ketoreductase (AFLD), which catalyzed the earliest and first stable aflatoxin precursor in aflatoxin biosynthetic pathway. AFLD was proved to be necessary for conversion of norsolorinic acid (NA) to averatin (AVN) in aflatoxin biosynthesis. For this reason, aflD was cloned and specific monoclonal antibodies for AFLD were developed to better define potential pathways of AFLD involved in Aflatoxin B_1 (AFB_1) biosynthesis. Monoclonal antibodies 2A8-1E10 and 2F11-3D8 were successfully screened out by immunized mouse. Immunoblot analysis revealed that 2A8-1E10 had high sensitivity and specificity to identify native AFLD protein in A. flavus with detection limit of 3 ng/mL. These results showed that it was suitable for quantitative detection of AFLD in A. flavus isolate. Further investigation revealed that aflatoxin accumulations of various A. flavus were not dependent on AFLD biosynthesis.Overall, this is the first report for AFLD monoclonal antibody development and application in A. flavus quantitative detection.
基金supported by the Public Welfare Application Research Project of Huzhou(No.2024GZ33)National Natural Science Foundation of China(No.31901799,32372417,22406069)+1 种基金Chinese Postdoctoral Science Foundation(No.2021M692370)National Key Research and Development Program of China(No.2022YFD2101203).
文摘The aflatoxin B1(AFB1)was considered as the most toxic natural compound and exists widely in the world,it is great significance to develop a high-performance sensor for simultaneously detection AFB1 and its toxin-producing gene aflD.In this paper,a magnetically controlled fluorescent sensor was developed,ssDNA1/ssDNA2 was modified on graphene quantum dots(GQDs)/Mn:ZnS quantum dots(QDs)as an energy donor,and cDNA/Apt modified by black hole quencher(BHQ)was coupled with MNPs as an energy acceptor,which leaded to fluorescence quenching through donor-acceptor energy transfer.Parts of fluorescent probes were separated from magnetic carriers when the sensor was incubated with the targets.After magnetic separation,the fluo-rescence intensity of the fluorescent probe released from the supernatant was positively correlated with the target concentration.The simultaneous detection of aflD and AFB1 was realized.The sensor displayed a wide detection range(0.1-500 nM and 0.1-500 ng/mL)for aflD and AFB1,with detection limits of 0.021 nM and 0.038 ng/mL for aflD and AFB1.The magnetic fluorescence biosensor showed a good recovery rate in real rice flour and corn flour samples.The magnetic control fluorescence biosensor had the high sensitivity and speci-ficity.This strategy provided a simple,efficient and sensitive method for food safety monitoring.
基金the Special Foundation for Science and Technology Basic Research Program(2019FY101103)the Natural Science Foundation of China(81772170,91846302,82073637,82003548)+5 种基金the National Key Research and Development Program of China(grant numbers:2017 YFC0907000,2017YFC0907500,2017YFC0211700,2019Y FC1315804)key basic research grants from the Science and Technology Commission of Shanghai Municipality(grant num-ber:16JC1400500)the Shanghai Municipal Science and Technology Major Project(No2017SHZDZX01)Three-Year Action Plan for Strengthening Public Health System in Shang-hai(grant number:GWV-10.2-YQ32)Innovation Grant from Science and Technology Commission of Shanghai Municipality,China(grant number:20ZR1405600)Local Innovative and Research Teams Project of Guangdong Pearl River Talents Pro-gram(2017BT01S131).
文摘Background and Aims:Metabolic dysfunction and obe-sity commonly coexist with both alcoholic and nonalcoholic fatty liver disease(AFLD and NAFLD).The association of AFLD and NAFLD with incident diseases in individuals with different metabolic phenotypes are unclear.Methods:UK Biobank study participants were screened for the presence of fatty liver at baseline.Body mass index and metabolic dysfunction were used to define metabolic phenotypes.Cox regression model was performed to examine the associations of AFLD and NAFLD with incident significant liver diseases(SLDs),cardiovascular diseases(CVDs),chronic kidney dis-eases(CKDs),and cancers,respectively.Results:A total of 43,974 AFLD and 103,248 NAFLD cases were identified.Both AFLD and NAFLD were associated with an increased risk of diseases of interest.The effects were amplified by obesity and metabolic abnormalities and modified by metabolic phe-notypes.Compared to individuals free of fatty liver and with phenotype of metabolically healthy-normal weight,AFLD[hazard ratio(HR 3.27;95%CI:1.95-5.47)]and NAFLD(HR 2.25;95%CI:1.28-3.94)cases with phenotype of met-abolically obese-normal weight had the greatest risk of SLDs.For CVDs,CKDs,and cancer,the greatest risks were detected in AFLD and NAFLD cases with phenotype of metabolically obese-overweight/obesity.In this subpopulation,AFLD and NAFLD conferred a 2.75-fold(95%CI:2.32-3.25)and 4.02-fold 95%CI:(3.64-4.43)increased risk of CVDs,4.37-fold 95%CI:(3.38-5.64)and 6.55-fold 95%CI:(5.73-7.48)increased risk of CKDs,and 1.19-fold 95%CI:(1.08-1.27)and 1.21-fold 95%CI:(1.14-1.28)increased risk of cancers,respectively.Conclusions:Metabolic phenotypes modified the association of AFLD and NAFLD with intrahepatic and ex-trahepatic diseases.
