In the present work,dispersive liquid-liquid microextraction(DLLME)was used to extract six synthetic cannabinoids(JWH-018,JWH-019,JWH-073,JWH-200,or WIN 55,225,JWH-250,and AM-694)from oral fluids.A rapid baseline sepa...In the present work,dispersive liquid-liquid microextraction(DLLME)was used to extract six synthetic cannabinoids(JWH-018,JWH-019,JWH-073,JWH-200,or WIN 55,225,JWH-250,and AM-694)from oral fluids.A rapid baseline separation of the analytes was achieved on a bidentate octadecyl silica hydride phase(Cogent Bidentate C18;4.6 mm×250 mm,4μm)maintained at 37℃,by eluting in isocratic conditions(water:acetonitrile(25:75,V/V)).Detection was performed using positive electrospray ionization-tandem mass spectrometry.The parameters affecting DLLME(pH and ionic strength of the aqueous phase,type and volume of the extractant and dispersive solvent,vortex and centrifugation time)were optimized for maximizing yields.In particular,using 0.5 mL of oral fluid,acetonitrile(1 mL),was identified as the best option,both as a solvent to precipitate proteins and as a dispersing solvent in the DLLME procedure.To select an extraction solvent,a low transition temperature mixture(LTTM;composed of sesamol and chlorine chloride with a molar ratio of 1:3)and dichloromethane were compared;the latter(100μL)was proved to be a better extractant,with recoveries ranging from 73%to 101%by vortexing for 2 min.The method was validated according to the guidelines of Food and Drug Administration bioanalytical methods:intra-day and inter-day precisions ranged between 4%and 18%depending on the spike level and analyte;limits of detection spanned from 2 to 18 ng/mL;matrixmatched calibration curves were characterized by determination coefficients greater than 0.9914.Finally,the extraction procedure was compared with previous methods and with innovative techniques,presenting superior reliability,rapidity,simplicity,inexpensiveness,and efficiency.展开更多
In all studies that analyzed biodiesel pollution effect of this compound on animals and especially marine animals. Biodiesel (methanol and sodium hydroxide) as to be taken into account. Less emphasis is placed on biod...In all studies that analyzed biodiesel pollution effect of this compound on animals and especially marine animals. Biodiesel (methanol and sodium hydroxide) as to be taken into account. Less emphasis is placed on biodiesel pollution of soils and waters (which can occur in the event of an environmental accident), which is considered a biodegradable product. But an accidental pollution due to the derailment of a freight train (August 12, 2020 on the CF line the Bucharest-Craiova highway, in the area of the CF bridge from Cârcea locality, Dolj county, Romania), confirmed the lack of knowledge in this field, as well as the way of depollution and restoration of the environment. This article describes how to pollute the soil with biodiesel, as well as the mathematical equations that describe this phenomenon.展开更多
Single-cell mass cytometry(SCMC)combines features of traditional flow cytometry(i.e.,fluorescence-activated cell sorting)with mass spectrometry,making it possible to measure several parameters at the single-cell level...Single-cell mass cytometry(SCMC)combines features of traditional flow cytometry(i.e.,fluorescence-activated cell sorting)with mass spectrometry,making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms.In this study,we optimized SCMC to analyze hemocytes of the Drosophila innate immune system.We used metal-conjugated antibodies(against cell surface antigens H2,H3,H18,L1,L4,and P1,and intracellular antigens 3A5 and L2)and anti-IgM(against cell surface antigen L6)to detect the levels of antigens,while anti-GFP was used to detect crystal cells in the immune-induced samples.We investigated the antigen expression profile of single cells and hemocyte populations in naive states,in immune-induced states,in tumorous mutants bearing a driver mutation in the Drosophila homologue of Janus kinase(hopTum)and carrying a deficiency of the tumor suppressor gene lethal(3)malignant blood neoplasm-1[l(3)mbn1],as well as in stem cell maintenance-defective hdcD84 mutant larvae.Multidimensional analysis enabled the discrimination of the functionally different major hemocyte subsets for lamellocytes,plasmatocytes,and crystal cells,and delineated the unique immunophenotype of Drosophila mutants.We have identified subpopulations of L2^(+)/P1^(+)and L2^(+)/L4^(+)/P1^(+)transitional phenotype cells in the tumorous strains l(3)mbn1 and hopTum,respectively,and a subpopulation of L4^(+)/P1^(+)cells upon immune induction.Our results demonstrated for the first time that SCMC,combined with multidimensional bioinformatic analysis,represents a versatile and powerful tool to deeply analyze the regulation of cell-mediated immunity of Drosophila.展开更多
基金supported by the Sapienza University of Rome through the project RICERCA 2019(protocol number:RG11916B6451D44A)。
文摘In the present work,dispersive liquid-liquid microextraction(DLLME)was used to extract six synthetic cannabinoids(JWH-018,JWH-019,JWH-073,JWH-200,or WIN 55,225,JWH-250,and AM-694)from oral fluids.A rapid baseline separation of the analytes was achieved on a bidentate octadecyl silica hydride phase(Cogent Bidentate C18;4.6 mm×250 mm,4μm)maintained at 37℃,by eluting in isocratic conditions(water:acetonitrile(25:75,V/V)).Detection was performed using positive electrospray ionization-tandem mass spectrometry.The parameters affecting DLLME(pH and ionic strength of the aqueous phase,type and volume of the extractant and dispersive solvent,vortex and centrifugation time)were optimized for maximizing yields.In particular,using 0.5 mL of oral fluid,acetonitrile(1 mL),was identified as the best option,both as a solvent to precipitate proteins and as a dispersing solvent in the DLLME procedure.To select an extraction solvent,a low transition temperature mixture(LTTM;composed of sesamol and chlorine chloride with a molar ratio of 1:3)and dichloromethane were compared;the latter(100μL)was proved to be a better extractant,with recoveries ranging from 73%to 101%by vortexing for 2 min.The method was validated according to the guidelines of Food and Drug Administration bioanalytical methods:intra-day and inter-day precisions ranged between 4%and 18%depending on the spike level and analyte;limits of detection spanned from 2 to 18 ng/mL;matrixmatched calibration curves were characterized by determination coefficients greater than 0.9914.Finally,the extraction procedure was compared with previous methods and with innovative techniques,presenting superior reliability,rapidity,simplicity,inexpensiveness,and efficiency.
文摘In all studies that analyzed biodiesel pollution effect of this compound on animals and especially marine animals. Biodiesel (methanol and sodium hydroxide) as to be taken into account. Less emphasis is placed on biodiesel pollution of soils and waters (which can occur in the event of an environmental accident), which is considered a biodegradable product. But an accidental pollution due to the derailment of a freight train (August 12, 2020 on the CF line the Bucharest-Craiova highway, in the area of the CF bridge from Cârcea locality, Dolj county, Romania), confirmed the lack of knowledge in this field, as well as the way of depollution and restoration of the environment. This article describes how to pollute the soil with biodiesel, as well as the mathematical equations that describe this phenomenon.
基金the National Research,Development and Innovation Office,Hungary(Grant Nos.GINOP-2.3.2-15-2016-00001,GINOP-2.3.2-15-2016-00030 to LGP,GINOP-2.3.2-15-2016-00035 to E´K,NKFI NN118207 and NKFI K120142 to IA,NKFI 120140 to EK,and OTKA K-131484 to VH)Gabor J.Szebeni was supported by the New National Excellence Program of the Ministry for Innovation and Technology,Hungary(Grant No.UNKP-19-4-SZTE-36)and by the Janos Bolyai Research Scholarship of the Hungarian Academy of Sciences(Grant No.BO/00139/17/8)。
文摘Single-cell mass cytometry(SCMC)combines features of traditional flow cytometry(i.e.,fluorescence-activated cell sorting)with mass spectrometry,making it possible to measure several parameters at the single-cell level for a complex analysis of biological regulatory mechanisms.In this study,we optimized SCMC to analyze hemocytes of the Drosophila innate immune system.We used metal-conjugated antibodies(against cell surface antigens H2,H3,H18,L1,L4,and P1,and intracellular antigens 3A5 and L2)and anti-IgM(against cell surface antigen L6)to detect the levels of antigens,while anti-GFP was used to detect crystal cells in the immune-induced samples.We investigated the antigen expression profile of single cells and hemocyte populations in naive states,in immune-induced states,in tumorous mutants bearing a driver mutation in the Drosophila homologue of Janus kinase(hopTum)and carrying a deficiency of the tumor suppressor gene lethal(3)malignant blood neoplasm-1[l(3)mbn1],as well as in stem cell maintenance-defective hdcD84 mutant larvae.Multidimensional analysis enabled the discrimination of the functionally different major hemocyte subsets for lamellocytes,plasmatocytes,and crystal cells,and delineated the unique immunophenotype of Drosophila mutants.We have identified subpopulations of L2^(+)/P1^(+)and L2^(+)/L4^(+)/P1^(+)transitional phenotype cells in the tumorous strains l(3)mbn1 and hopTum,respectively,and a subpopulation of L4^(+)/P1^(+)cells upon immune induction.Our results demonstrated for the first time that SCMC,combined with multidimensional bioinformatic analysis,represents a versatile and powerful tool to deeply analyze the regulation of cell-mediated immunity of Drosophila.
