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In this work, a vortex-assisted dispersive liquid-liquid microextraction method, using an ionic liquid as the extracting solvent was developed, for the simultaneous analysis of three UV filters in different water samples. The extracting and dispersive solvents were selected in a univariate way. Then, the parameters such as the volume of the extracting and dispersive solvents, pH and ionic strength were evaluated using a full experimental design 24, followed by Doehlert matrix. The optimized method consisted of 50 µL of extracting solvent (1-octyl-3-methylimidazolium hexafluorophosphate), 700 µL of dispersive solvent (acetonitrile) and pH of 4.5. When combined with high-performance liquid chromatography, the method limit of detection ranged from 0.3 to 0.6 µg L-1, enrichment factors between 81 and 101%, and the relative standard deviation between 5.8 and 10.0%. The developed method demonstrated effectiveness in concentrating UV filters in both river and seawater samples, being a simple and efficient option for this type of analysis.
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In the context of classical molecular simulations, the accuracy of a force field is highly influenced by the values of the relevant simulation parameters. In this work, a parameter-space mapping (PSM) workflow is proposed to aid in the calibration of force-field parameters, based mainly on the following features: (i) regular-grid discretization of the search space; (ii) partial sampling of the search-space grid; (iii) training of surrogate models to predict the estimates of the target properties for nonsampled parameter sets; (iv) post hoc interpretation of the results in terms of multiobjective optimization concepts; (v) attenuation of statistical errors achieved via empiric extension of the duration of the simulations; (vi) iterative search-space translation according to a user-defined scalar objective function that measures the accuracy of the force field (e.g., the weighted root-mean-square deviation of the target properties relative to the reference data). This combination of features results in a hybrid of a single- and a multiobjective optimization strategy, allowing for the approximate determination of both a local minimum of the chosen objective function and its neighboring Pareto efficient points. The PSM workflow is implemented in the extensible Python program gmak, which is made available in the Git repository at http://github.com/mssm-labmmol/gmak. Using this implementation, the PSM workflow was tested in a proof-of-concept fashion in the recalibration of the Lennard-Jones parameters of the 3-point Optimal Point Charge (OPC3) water model for compatibility with the GROMOS treatment of nonbonded interactions. The recalibrated model reproduces typical pure-liquid properties with an accuracy similar to the original OPC3 model and represents a significant improvement relative to the Simple Point Charge (SPC) model, which is the official recommendation for simulations using GROMOS force fields.
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The calibration of torsional interaction terms by fitting relative gas-phase conformational energies against their quantum-mechanical values is a common procedure in force-field development. However, much less attention has been paid to the optimization of third-neighbor nonbonded interaction parameters, despite their strong coupling with the torsions. This article introduces an algorithm termed LLS-SC, aimed at simultaneously parametrizing torsional and third-neighbor interaction terms based on relative conformational energies. It relies on a self-consistent (SC) procedure where each iteration involves a linear least-squares (LLS) regression followed by a geometry optimization of the reference structures. As a proof-of-principle, this method is applied to obtain torsional and third-neighbor interaction parameters for aliphatic chains in the context of the GROMOS 53A6 united-atom force field. The optimized parameter set is compared to the original one, which has been fitted manually against thermodynamic properties for small linear alkanes. The LLS-SC implementation is freely available under http://github.com/mssm-labmmol/profiler.
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Hyptis dilatata and Mesosphaerum suaveolens belong to Lamiaceae, are aromatic and medicinal subshrubs with antiparasitic potential and occurrence in the Amazon Region. The essential oils of both species were extracted, analyzed, and identified by GC and GC-MS and then evaluated their anthelmintic activities against the nematode Caenorhabditis elegans. Primary constituents of the samples of H. dilatata oils were limonene (72.6%), myrcene (11.5%), and p-cymene (10.3%) to PAMA19 sample, and camphor (25.5%), α-pinene (25.4%), 1,8-cineole (18.8%), ß-pinene (12.0%), and limonene (5.9%) to PAMA108 sample, while in the oil of M. suaveolens, PAMA131 sample, predominated bicyclogermacrene (23.5%, 1,8-cineole (23.0%), germacrene D (17.2%), and (E)-caryophyllene (10.4%). The sample oil of H. dilatata (PAMA108) exhibited the lower anthelmintic inhibitory concentration, with an IC50 value of 2.09 mg/mL for C. elegans Bristol N2 strain, while the oils of H. dilatata (PAMA19) and M. suaveolens (PAMA131) showed an IC50 up of 4 mg/mL for C. elegans IVR15 and Bristol N2 strains. These results suggest that the H. dilatata and M. suaveolens oils constituents' combination can be helpful as a nematicidal agent due to their synergistic action.
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Anti-Helmínticos , Hyptis , Óleos Voláteis , Animais , Anti-Helmínticos/farmacologia , Caenorhabditis elegans , Cromatografia Gasosa-Espectrometria de Massas , Óleos Voláteis/farmacologia , Óleos de Plantas/farmacologiaRESUMO
The effect of different treatments of the nonbonded interactions in simulations employing the recently introduced GROMOS-compatible 2016H66 force field is evaluated based on calculations carried out with the GROMACS software. This is done considering four thermodynamic and transport properties (pure liquid density, vaporization enthalpy, surface-tension coefficient, and self-diffusion constant) of 58 organic liquids representative of the chemical groups alcohol, ether, aldehyde, ketone, carboxylic acid, ester, amine, amide, thiol, sulfide, disulfide, and aromatic compounds, also including water (SPC model). A dipalmitoylphosphatidylcholine bilayer system is considered as well. The simulated properties are found to be very sensitive to the treatment of the long-range dispersion interactions, notably for the least polar systems. In general, the treatment of the long-range electrostatic or Lennard-Jones interactions using homogeneous correction terms or lattice-sum approaches yield similar results, with punctual discrepancies. The combination of a lattice-sum approach for the electrostatic interactions with a straight-cutoff truncation of the Lennard-Jones interactions at a distance of at least 1.2 nm is found to represent a good compromise setup within GROMACS for achieving compatibility with the reference results obtained using GROMOS as well as a comparable level of agreement with the experimental data. This study also reveals two potential issues with the GROMACS software, related to an incorrect calculation of the pressure when using LINCS in version 4.0.7 and an inadequate implementation of the twin-range scheme in version 5.1.2.
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The development of age-related cardiovascular disease is associated with the senescence of vascular cells. This study aimed to investigate the effect of ginsenoside Rg1 on vascular smooth muscle cell (VSMC) senescence. Primary VSMCs were cultured and divided into control, D-galactose (D-gal), Rg1-L, and Rg1-H groups, which were cultured without and with D-gal, and with low- and high-concentrations of Rg1, respectively. D-gal-induced cellular senescence was identified by b-galactosidase staining, and ultrastructural changes within the cells were observed. The expression of p16, p21, and p53 in the four groups of VSMCs was determined by western blotting, and the cell cycle was investigated by flow cytometry. Compared with the control group, there was an obvious change in the ultrastructure of VSMCs in the D-gal group, and the proportion of b-galactosidase-positive cells was significantly increased (P < 0.05). In addition, p16, p21, and p53 expression was significantly increased (P < 0.05) and the cell cycle was arrested in the G0/G1 phase. Compared with the D-gal group, the percentage of positive cells was significantly reduced (P < 0.05) in the Rg1 groups, the expression of p16, p21, and p53 was significantly reduced (P < 0.05), and the number of cells in the G0/G1 phase decreased (P < 0.05). Ginsenoside Rg1 can inhibit VSMC senescence, and the mechanisms may be related to its partial inhibition of the p16INK4a/Rb and p53-p21Cip1/Waf1 signaling pathways during the cell cycle.
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Senescência Celular/efeitos dos fármacos , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Ginsenosídeos/farmacologia , Miócitos de Músculo Liso/efeitos dos fármacos , Animais , Adesão Celular/efeitos dos fármacos , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/genética , Proliferação de Células/efeitos dos fármacos , Inibidor p16 de Quinase Dependente de Ciclina/genética , Inibidor p16 de Quinase Dependente de Ciclina/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/genética , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , Medicamentos de Ervas Chinesas/química , Galactose/farmacologia , Regulação da Expressão Gênica , Ginsenosídeos/isolamento & purificação , Masculino , Músculo Liso Vascular/citologia , Músculo Liso Vascular/efeitos dos fármacos , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/citologia , Miócitos de Músculo Liso/metabolismo , Cultura Primária de Células , Ratos , Ratos Sprague-Dawley , Transdução de Sinais , Proteína Supressora de Tumor p53/genética , Proteína Supressora de Tumor p53/metabolismo , beta-Galactosidase/genética , beta-Galactosidase/metabolismoRESUMO
Pseudomembranous colitis (PMC) occurs mainly in adults and is believed to be caused almost exclusively by toxins produced by Clostridium difficile. Colonoscopy found that PMC occurs mainly in the colon, sigmoid colon and rectum in up to 80% ~ 100% of cases. Colonoscopy is simple and fast. It has the significance of making a definite diagnosis and can be used as the main examination method of diagnosis. Reports of children suffering from PMC are rare. Herein, we report a case of PMC in a child. This report has some clinical value for the study of the spectrum of PMC in patients.
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The association of single nucleotide polymorphisms (SNPs) in PPARγ with hypertension is controversial. The aim of the present study was to clarify the contributions of PPARγ genetic variants to hypertension through an association study. A total of 414 unrelated Mongolian herdsmen and 524 Han farmers were included in this study. Fourteen intronic SNPs were analyzed and genotyped using a polymerase chain reaction/ligase detection reaction assay. Prior to correction for multiple testing, the SNPs rs6802898 and rs12633551 were significantly associated with the prevalence of hypertension in the Han and Mongolian populations, respectively. The genetic association of each SNP with hypertension was individually tested using logistic regression. The SNP rs6802898 was associated with hypertension in both dominant (P = 0.033) and additive models (P = 0.026) in the Han population, whereas the SNP rs12633551 was associated with hypertension in both dominant (P = 0.014) and additive models (P = 0.0073) in the Mongolian population. Moreover, SNP rs12633551 had a significant effect on systolic and diastolic blood pressure response. However, none of these associations were statistically significant after Bonferroni correction for multiple testing, although there was a significant difference among the haplotypes in the Han and Mongolian populations. Interestingly, there was an association of the PPARγ haplotypes with hypertension even after Bonferroni correction. Thus, determination of the PPARγ haplotypes in different populations may prove informative for assessment of the genetic risk for hypertension.
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Predisposição Genética para Doença , Hipertensão/genética , Modelos Genéticos , PPAR gama/genética , Polimorfismo de Nucleotídeo Único , Adulto , Idoso , Alelos , Povo Asiático , Pressão Sanguínea , China/epidemiologia , Éxons , Fazendeiros , Feminino , Expressão Gênica , Frequência do Gene , Estudo de Associação Genômica Ampla , Haplótipos , Humanos , Hipertensão/diagnóstico , Hipertensão/epidemiologia , Hipertensão/fisiopatologia , Íntrons , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , PrevalênciaRESUMO
Twenty-four microsatellite markers were isolated from the genomic DNA of Amomum tsaoko Crevost et Lemaire, an important economic plant in China, using the method fast isolation by AFLP of sequences containing repeats (FIASCO). Polymorphism within each locus was assessed in 60 individuals from three populations in Yunnan Province, China, and nine of them were polymorphic. The number of alleles per polymorphic locus was 2, and the expected and observed heterozygosities ranged from 0.224 to 0.513, and from 0.050 to 0.600, respectively. Among nine microsatellite markers with polymorphism, five showed significant deviation from Hardy-Weinberg equilibrium (P < 0.01), probably due to anthropic selection and short-cloning history in cultivation. No significant linkage disequilibrium was detected between loci in our analysis. These polymorphic microsatellite markers will facilitate further studies of gene flow, population structure, identification of cultivated variety, and evaluation of germplasm resources.
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Repetições de Microssatélites , Zingiberaceae/genética , China , Loci Gênicos , Polimorfismo GenéticoRESUMO
We investigated the reversal effect of vitamin D on the multidrug-resistant leukemic Jurkat/ADR and K562/ADR cell lines and conducted a preliminary investigation of its reversal mechanism. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method was used to detect the reversal effect of vitamin D on multidrug-resistant cells. Real-time polymerase chain reaction was used to determine the effect of vitamin D on intracellular expression of mRNA of the multidrug-resistant gene (MDRI) and the multidrug-resistance-related gene (MRP1). A protein quantitative analysis method was used to determine the effect of vitamin D on intracellular glutathione content. After treatment of Jurkat/ADR and K562/ADR cells with vitamin D, multidrug resistance was reversed in a dose-dependent manner, which may have reduced mRNA expression of the MDR1 and MRP1 genes, the P-glycoprotein content on the cell surface, and the intracellular glutathione level. Different concentrations of vitamin D showed varying reversal effects on different multidrug-resistant cells. The resistance mechanism may be related to the inhibition of the expression of MDR1 and MRP1 genes.
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Antineoplásicos/farmacologia , Resistencia a Medicamentos Antineoplásicos/efeitos dos fármacos , Regulação Leucêmica da Expressão Gênica , Proteínas Associadas à Resistência a Múltiplos Medicamentos/antagonistas & inibidores , RNA Mensageiro/antagonistas & inibidores , Vitamina D/farmacologia , Subfamília B de Transportador de Cassetes de Ligação de ATP/antagonistas & inibidores , Subfamília B de Transportador de Cassetes de Ligação de ATP/genética , Subfamília B de Transportador de Cassetes de Ligação de ATP/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Citarabina/farmacologia , Doxorrubicina/farmacologia , Resistencia a Medicamentos Antineoplásicos/genética , Glutationa/metabolismo , Humanos , Concentração Inibidora 50 , Células Jurkat , Células K562 , Proteínas Associadas à Resistência a Múltiplos Medicamentos/genética , Proteínas Associadas à Resistência a Múltiplos Medicamentos/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais , Vincristina/farmacologiaRESUMO
Chronic obstructive pulmonary disease (COPD) is a progressive lung disease characterized by persistent airflow limitation. Smoking, occupational exposures, air pollution, and genetics are all risk factors. In the present study, we detected the cytochrome P4501A1 gene (CYP1A1) MspI polymorphism and the tumor necrosis factor alpha (TNFα)-308 single nucleotide polymorphism in COPD patients, and investigated their associations with smoking and COPD susceptibility in Inner Mongolia. A total of 101 COPD patients and 80 controls were enrolled in the study. The polymorphisms were analyzed using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). CYP1A1 MspI allele frequencies were significantly different between COPD patients and controls (P=0.033). COPD susceptibility was higher in subjects with the m2 allele compared to subjects with the m1 allele [odds ratio (OR)=2.531, 95% confidence interval (CI)=1.297-4.940, P=0.006]. Significant differences were observed in the TNFα-308 genotype and allele distributions between COPD patients and controls (P=0.006 and P=0.003, respectively). Compared to subjects with the GG genotype, subjects with GA+AA genotypes had higher COPD risk (OR=3.639, 95%CI=1.576-8.403, P=0.002). The TNFα-308 polymorphism differed between smoking and non-smoking COPD patients and controls (P=0.047 for genotype and P=0.030 for allele). In conclusion, the CYP1A1 MspI and TNFα-308 polymorphisms were associated with COPD susceptibility. Furthermore, of the two polymorphisms, only TNFα-308 may exert an interaction with smoking.
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Citocromo P-450 CYP1A1/genética , Doença Pulmonar Obstrutiva Crônica/genética , Fator de Necrose Tumoral alfa/genética , Idoso , China , Feminino , Frequência do Gene , Estudos de Associação Genética , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Polimorfismo de Nucleotídeo Único , Doença Pulmonar Obstrutiva Crônica/patologia , Fumar/efeitos adversosRESUMO
Lifestyle, habits, diet, and genetics are all important factors associated with the prevalence of hypertension. Many association studies have been performed in the Chinese Han population, whereas data explaining the high prevalence of hypertension in the Mongolian population remain scarce. In the present study, we aimed to determine the factors associated with the development of essential hypertension in Mongolians. A total of 194 hypertensive cases and 201 controls from Dongwu County were enrolled in the study. Demographics, anthropometric and blood biochemical parameters, food intake, lifestyle, habits, education, occupation, and family history were recorded for each subject. Genotype and allele frequencies of six single nucleotide polymorphisms (SNPs) of the kallikrein 1 (KLK1) gene were also examined. Mean body mass index, waistline, hipline, blood sugar, blood urea nitrogen, creatinine, uric acid, total cholesterol, triglyceride, and low-density lipoprotein levels were all significantly higher in the hypertensive group (P<0.01). Hypertensives consumed less milk, vegetables, and fruits, and had higher cigarette, alcohol, and salt intake (P<0.05). There were also less regular physical exercisers and manual workers among the hypertensive group (P<0.05). The mean inheritance rank of the hypertensive group was higher than that of controls (P<0.05). There were no differences in the distribution of genotype and allele frequencies of the six SNPs between the hypertensive and control groups (P>0.05). These results suggest that dietary history and habits have the most important influence on the development of essential hypertension in the Mongolian population.
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Pressão Sanguínea/genética , Hipertensão/genética , Calicreínas/genética , Adulto , Índice de Massa Corporal , Estudos de Casos e Controles , China , Dieta , Hipertensão Essencial , Etnicidade/genética , Feminino , Estudos de Associação Genética , Humanos , Hipertensão/sangue , Hipertensão/patologia , Estilo de Vida , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Triglicerídeos/sangueRESUMO
Human cytochrome P450 4A11 (CYP4A11) plays a role in the regulation of blood pressure through the conversion of arachidonic acid into 20-hydroxyeicosatetraenoic acid (20-HETE). We therefore investigated the association between a CYP4A11 polymorphism (rs9333025) with hypertension in the Mongolian and Han ethnic groups. We studied 514 Mongolians in a pastoral area, including 201 hypertension patients and 313 normotensive controls, and 524 Han individuals in an urban area, including 215 hypertension patients and 309 normotensive controls. Genotyping was performed using polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Genotype, allele, and dominant inheritance differed significantly between the Mongolian and Han populations (P = 0.006, P = 0.002, and P = 0.003, respectively). Significant differences were also observed in these factors when considering only males (P = 0.001, P = 0.003, and P = 0.001, respectively). For the Han population, recessive inheritance differed significantly between hypertension patients and controls and between male patients and controls (P = 0.005 and P = 0.049, respectively). The genotypic, allelic, and dominant frequencies differed significantly between hypertension patients in both populations (P = 0.019, P = 0.035, and P = 0.024, respectively). The genotypic frequency in Mongolian male patients was significantly different from that in Han male patients (P = 0.009). Higher body mass index, triglycerides, and lower high-density lipoprotein were associated with increased risk of developing hypertension in the Han population. The GG genotype was in higher frequency in the Mongolian population, indicating that it is a high risk factor for hypertension. Mongolian men were at higher risk of developing hypertension.
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Sistema Enzimático do Citocromo P-450/genética , Hipertensão/genética , Polimorfismo de Fragmento de Restrição , Adulto , Idoso , Povo Asiático , Estudos de Casos e Controles , China , Citocromo P-450 CYP4A , Feminino , Estudos de Associação Genética , Humanos , Hipertensão/epidemiologia , Hipertensão/etnologia , Masculino , Pessoa de Meia-Idade , Mongólia/etnologia , Fatores SexuaisRESUMO
Activation of the peroxisome proliferator-activated receptor g (PPARg) improves insulin sensitivity and inhibits atherosclerosis. Whether PPARg2 Pro12Ala polymorphism affects myocardial infarction is not clearly understood. We investigated a possible association of PPARg2 Pro12Ala polymorphism with obesity and myocardial infarction in Han Chinese in Hohhot, Inner Mongolia, China. We included 121 subjects with myocardial infarction and 137 healthy controls in our study. Triglycerides, total cholesterol, low-density lipoprotein cholesterol, and high-density lipoprotein cholesterol were measured. The following information was recorded for each subject: age, gender, body height, body weight, systolic blood pressure, and diastolic blood pressure; the body mass index was calculated. PCR-RFLP was used to examine Pro12Ala polymorphism. There were significant differences in clinical characteristics between myocardial infarction patients and healthy controls, except for diastolic blood pressure and triglycerides. The PP, PA/AA genotype frequencies were 88.4 and 11.6% in myocardial infarction patients and 95.6 and 4.4% in controls, respectively (P = 0.031). Individuals with the A allele had a significantly higher risk of myocardial infarction. The A allele was not an independent risk factor for obesity. We conclude that PPARg2 Pro12Ala polymorphisms are associated with increased risk for myocardial infarction in Han Chinese in Hohhot.
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Povo Asiático/genética , Estudos de Associação Genética , Predisposição Genética para Doença , Infarto do Miocárdio/genética , Obesidade/genética , PPAR gama/genética , Polimorfismo de Nucleotídeo Único/genética , Idoso , Idoso de 80 Anos ou mais , Substituição de Aminoácidos/genética , Estudos de Casos e Controles , China , Etnicidade/genética , Feminino , Frequência do Gene/genética , Humanos , Modelos Logísticos , Masculino , Pessoa de Meia-Idade , Análise Multivariada , Infarto do Miocárdio/complicações , Obesidade/complicações , Fatores de RiscoRESUMO
We investigated a possible association of peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PGC-1α) Gly482Ser polymorphism with hypertension in Mongolians in Inner Mongolia. A total of 787 subjects were enrolled randomly, including 390 hypertension patients and 397 healthy controls. Triglycerides, cholesterol, and fasting plasma glucose were measured, and body mass index was calculated. PCR-RFLP was used to analyze Gly482Ser polymorphisms. There were significant differences in triglycerides, fasting plasma glucose, and body mass index between hypertension patients and healthy controls. Cholesterol levels did not differ significantly. The PGC-1α gene GG, GA and AA genotype distributions were 37.2, 48.5 and 14.4%, respectively, in patients and 48.6, 37.3 and 14.1% in healthy controls. The frequencies of PGC-1α genotype GA and allele A were significantly different between hypertension patients and healthy Mongolians. We concluded that PGC-1α Gly482Ser polymorphism is associated with hypertension in Mongolians in Inner Mongolia.