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1.
BMC Plant Biol ; 24(1): 170, 2024 Mar 05.
Artigo em Inglês | MEDLINE | ID: mdl-38443797

RESUMO

BACKGROUND: Panax notoginseng (Burk) F. H. Chen is one of the most famous Chinese traditional medicinal plants. The taproot is the main organ producing triterpenoid saponins, and its development is directly linked to the quality and yield of the harvested P. notoginseng. However, the mechanisms underlying the dynamic metabolic changes occurring during taproot development of P. notoginseng are unknown. RESULTS: We carried out metabolomic and transcriptomic analyses to investigate metabolites and gene expression during the development of P. notoginseng taproots. The differentially accumulated metabolites included amino acids and derivatives, nucleotides and derivatives, and lipids in 1-year-old taproots, flavonoids and terpenoids in 2- and 3-year-old taproots, and phenolic acids in 3-year-old taproots. The differentially expressed genes (DEGs) are related to phenylpropanoid biosynthesis, metabolic pathway and biosynthesis of secondary metabolites at all three developmental stages. Integrative analysis revealed that the phenylpropanoid biosynthesis pathway was involved in not only the development of but also metabolic changes in P. notoginseng taproots. Moreover, significant accumulation of triterpenoid saponins in 2- and 3-year-old taproots was highly correlated with the up-regulated expression of cytochrome P450s and uridine diphosphate-dependent glycosyltransferases genes. Additionally, a gene encoding RNase-like major storage protein was identified to play a dual role in the development of P. notoginseng taproots and their triterpenoid saponins synthesis. CONCLUSIONS: These results elucidate the molecular mechanism underlying the accumulation of and change relationship between primary and secondary metabolites in P. notoginseng taproots, and provide a basis for the quality control and genetic improvement of P. notoginseng.


Assuntos
Panax notoginseng , Saponinas , Triterpenos , Panax notoginseng/genética , Metaboloma , Perfilação da Expressão Gênica
2.
Front Psychiatry ; 14: 1138205, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37032928

RESUMO

Objective: Major depressive disorder (MDD) has a relapse rate that cannot be ignored and places a tremendous burden on the patient in the prevention and treatment process. Yoga, a combination of physical and mental exercises, is effective and acceptable for the adjunctive treatment of MDD. This study aimed to explore further the evidence of yoga's efficacy for patients with MDD. Methods: PubMed, Embase, Cochrane library, PsycINFO, SinoMed, CNKI, Wanfang, and VIP databases from their inception to 13 October 2022 were searched by a pre-defined search strategy. RCTs of patients with MDD who met diagnostic criteria for yoga treatment were included. RoB2.0 was used to evaluate the quality of the literature. Improvement in depressive symptoms was assessed by the Beck Depression Inventory (BDI), Hamilton Depression Rating Scale (HAMD), or other scales were used as primary outcome indicators, and improvement in anxiety was assessed by the Hamilton Anxiety Scale (HAMA) and State-Trait Anxiety Inventory (STAI) scale as secondary outcome indicators. RR and Cohen's d at 95% CI were used as effect size estimates, and Q and I2 were used to evaluate the size of heterogeneity, with a p-value less than 0.05 indicating statistical significance. Results: Thirty-four RCT studies, including 1,269 patients in the treatment group and 1,072 patients in the control group, 48.4% of whom were women, were included in the study. Compared to the control group, the BDI-II results yielded a moderate effect of yoga on the improvement of depressive symptoms (Cohen's d = -0.60; 95% CI: -1.00 to -0.21; p < 0.01), the HAMD results yielded a moderate improvement of yoga on the severity of depressive symptoms (Cohen's d = -0.64; 95% CI: -0.98 to -0.30; p < 0.01), and the STAI results can be concluded that yoga had a negligible effect on the improvement of the level of anxiety (Cohen's d = -0.26; 95% CI: -0.48 to -0.04; p = 0.02). No adverse events occurred in the yoga group during the treatment. Conclusion: Yoga can improve depressive symptoms and anxiety in patients with MDD and has a safe and wide patient acceptance. Systematic review registration: [https://www.crd.york.ac.uk/prospero/], identifier [PROSPERO, CRD42022373282].

3.
Sci Rep ; 13(1): 4721, 2023 03 23.
Artigo em Inglês | MEDLINE | ID: mdl-36959272

RESUMO

Influences of photosynthesis of leaf and silique on seeds yield and quality of oilseed rape (Brassica napus L.) were explored in this study. A field comparing experiment with several rapeseed varieties was conducted and the results showed, that the leaf area index (LAI), silique surface area index (SAI), siliques number per plant, and biological yield were statistically classified as the first principal factors which greatly influenced seeds yield, the leaf net photosynthetic rate (Pn) and silique Pn were the second principal factors; the stomatal conductance (Gs) and chlorophyll a (Chl a) content were the first principal factors which influenced leaf Pn and silique Pn. A shading experiment was conducted and the results showed that, under treatments of the ZH1, ZH2, and ZH3 (shading rapeseed plants during flowering stage, during time from initial flowering until seeds ripening, and during time from flowering ending until seeds ripening, respectively), respectively the seeds yield per plant was reduced by 34.6%, 84.3%, and 86.1%, the seed protein content was significantly increased. The treatment ZH1 Not, but the ZH2 and ZH3 caused significant decrease in both seed oil content and oleic acid (C18:1) content in seed oil, and the contents of linoleic acid (C18:2), linolenic acid (C18:3) in oil were significantly increased, gene expression of the ACCase (Acetyl-CoA carboxylase), FAD2 (fatty acid desaturase), and FAD3 (ω-3 fatty acid dehydrogenase) in green seeds was restrained/changed. Thus the LAI, SAI, siliques number per plant, biological yield per plant, leaf Pn, silique Pn, and the Gs, Chl a content of leaf and silique formed an indexes system to be used in screening rapeseed variety with higher light efficiency and seeds yield; the silique photosynthesis inhibition and the photosynthates deficiency in rapeseed plant after flowering stage predominately influenced seeds yield and quality.


Assuntos
Brassica napus , Brassica rapa , Brassica napus/genética , Brassica napus/metabolismo , Clorofila A/metabolismo , Sementes/genética , Folhas de Planta , Fotossíntese/fisiologia , Óleos de Plantas/metabolismo
4.
Environ Geochem Health ; 45(12): 9029-9040, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36183309

RESUMO

Yunnan Province is the main planting area of the precious Chinese herbal medicines (CHM) Panax notoginseng; however, it locates the geological area with high soil heavy metals in China. The frequent land replacement due to continuous cropping obstacles and excessive application of chemicals makes P. notoginseng prone to be contaminated by heavy metals under the farmland P. notoginseng (FPn) planting. To overcome farmland shortage, understory P. notoginseng (UPn) was developed as a new ecological planting model featured by no chemicals input. However, this newly developed planting system requires urgently the soil-plant heavy metal characteristics and risk assessment. This study aimed to evaluate the pollution status of eight heavy metals in the tillage layer (0-20 cm), subsoil layer (20-40 cm) and the plants of UPn in Lancang County, Yunnan Province. Pollution index (Pi) showed that the contamination degree of heavy metals in the tillage layer and subsoil layer was Cd > Pb > Ni > Cu > Zn > Cr > Hg > As and Pb > Cd > Cu > Ni > Cr > Hg > Zn > As, respectively. Potential ecological risk index (PERI) for the tillage layer and subsoil layer was slight and middle, respectively. The exceeding standard rate of Cd, As, Pb, Hg, Cu in the UPn roots was 5.33%, 5.33%, 13.33%, 26.67% and 1.33%, respectively, while only Cd and Hg in the UPn leaves exceeded the standard 10% and 14%, respectively. The enrichment abilities of Cd and Hg in the roots and leaves of UPn were the strongest, while that of Pb was the weakest. The Hazard index (HI) and target hazard quotient (THQ) of eight heavy metals in the roots and leaves of UPn were less than 1.Therefore, our results prove that Upn has no human health risk and provide a scientific basis for the safety evaluation and extension of UPn.


Assuntos
Mercúrio , Metais Pesados , Panax notoginseng , Poluentes do Solo , Cádmio , Chumbo , Monitoramento Ambiental/métodos , China , Poluentes do Solo/análise , Metais Pesados/análise , Solo , Medição de Risco
5.
J Med Virol ; 95(1): e28438, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36573423

RESUMO

Coronavirus disease 2019 (COVID-19), as well as its prevention and control measures, seriously affected people's livehood, which may have affected the body's level of vitamin D (VD). This study aimed to investigate the effect of the COVID-19 pandemic on the VD status of children in Zhengzhou, China. In this study, we included 12 272 children in 2019 (before the COVID-19 pandemic) and 16 495 children in 2020 (during the COVID-19 pandemic) to examine the changes in VD levels and deficiency rates among children before and during the COVID-19 pandemic. Total VD levels in 2020 were significantly higher than those in 2019 (26.56 [18.15, 41.40] vs. 25.98 [17.92, 40.09] ng/ml, p < 0.001). Further analysis revealed that during the COVID-19 pandemic control period in 2020, the VD levels in February, March, and April were lower than those in the same months of 2019, while the VD deficiency rates were significantly higher. Additionally, our data revealed that VD levels decreased significantly with age. Among children older than 6 years, the VD deficiency rate exceeded 50%. These results indicate that we should pay close attention to VD supplementation during the COVID-19 pandemic control period and in children older than 6 years of age.


Assuntos
COVID-19 , Deficiência de Vitamina D , Criança , Humanos , Vitamina D , Estudos Transversais , Pandemias , COVID-19/epidemiologia , Vitaminas , Deficiência de Vitamina D/epidemiologia
6.
J Neural Eng ; 18(2)2021 02 24.
Artigo em Inglês | MEDLINE | ID: mdl-33395676

RESUMO

Objective. Motor imagery (MI) electroencephalography (EEG) classification is regarded as a promising technology for brain-computer interface (BCI) systems, which help people to communicate with the outside world using neural activities. However, decoding human intent accurately is a challenging task because of its small signal-to-noise ratio and non-stationary characteristics. Methods that directly extract features from raw EEG signals ignores key frequency domain information. One of the challenges in MI classification tasks is finding a way to supplement the frequency domain information ignored by the raw EEG signal.Approach. In this study, we fuse different models using their complementary characteristics to develop a multiscale space-time-frequency feature-guided multitask learning convolutional neural network (CNN) architecture. The proposed method consists of four modules: the space-time feature-based representation module, time-frequency feature-based representation module, multimodal fused feature-guided generation module, and classification module. The proposed framework is based on multitask learning. The four modules are trained using three tasks simultaneously and jointly optimized.Results. The proposed method is evaluated using three public challenge datasets. Through quantitative analysis, we demonstrate that our proposed method outperforms most state-of-the-art machine learning and deep learning techniques for EEG classification, thereby demonstrating the robustness and effectiveness of our method. Moreover, the proposed method is employed to realize control of robot based on EEG signal, verifying its feasibility in real-time applications.Significance. To the best of our knowledge, a deep CNN architecture that fuses different input cases, which have complementary characteristics, has not been applied to BCI tasks. Because of the interaction of the three tasks in the multitask learning architecture, our method can improve the generalization and accuracy of subject-dependent and subject-independent methods with limited annotated data.


Assuntos
Interfaces Cérebro-Computador , Algoritmos , Eletroencefalografia/métodos , Humanos , Imaginação , Redes Neurais de Computação
7.
Mol Ecol Resour ; 21(1): 153-169, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32985109

RESUMO

Erigeron breviscapus is an important medicinal plant in Compositae and the first species to realize the whole process from the decoding of the draft genome sequence to scutellarin biosynthesis in yeast. However, the previous low-quality genome assembly has hindered the optimization of candidate genes involved in scutellarin synthesis and the development of molecular-assisted breeding based on the genome. Here, the E. breviscapus genome was updated using PacBio RSII sequencing data and Hi-C data, and increased in size from 1.2 Gb to 1.43 Gb, with a scaffold N50 of 156.82 Mb and contig N50 of 140.95 kb, and a total of 43,514 protein-coding genes were obtained and oriented onto nine pseudo-chromosomes, thus becoming the third plant species assembled to chromosome level after sunflower and lettuce in Compositae. Fourteen genes with evidence for positive selection were identified and found to be related to leaf morphology, flowering and secondary metabolism. The number of genes in some gene families involved in flavonoid biosynthesis in E. breviscapus have been significantly expanded. In particular, additional candidate genes involved in scutellarin biosynthesis, such as flavonoid-7-O-glucuronosyltransferase genes (F7GATs) were identified using updated genome. In addition, three candidate genes encoding indole-3-pyruvate monooxygenase YUCCA2 (YUC2), serine carboxypeptidase-like 18 (SCPL18), and F-box protein (FBP), respectively, were identified to be probably related to leaf development and flowering by resequencing 99 individuals. These results provided a substantial genetic basis for improving agronomic and quality traits of E. breviscapus, and provided a platform for improving other draft genome assemblies to chromosome-level.


Assuntos
Erigeron , Genoma de Planta , Asteraceae , Erigeron/genética , Plantas Medicinais/genética
8.
BMC Plant Biol ; 19(1): 451, 2019 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-31655543

RESUMO

BACKGROUND: Taproot thickening is a complex biological process that is dependent on the coordinated expression of genes controlled by both environmental and developmental factors. Panax notoginseng is an important Chinese medicinal herb that is characterized by an enlarged taproot as the main organ of saponin accumulation. However, the molecular mechanisms of taproot enlargement are poorly understood. RESULTS: A total of 29,957 differentially expressed genes (DEGs) were identified during the thickening process in the taproots of P. notoginseng. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment revealed that DEGs associated with "plant hormone signal transduction," "starch and sucrose metabolism," and "phenylpropanoid biosynthesis" were predominantly enriched. Further analysis identified some critical genes (e.g., RNase-like major storage protein, DA1-related protein, and Starch branching enzyme I) and metabolites (e.g., sucrose, glucose, fructose, malate, and arginine) that potentially control taproot thickening. Several aspects including hormone crosstalk, transcriptional regulation, homeostatic regulation between sugar and starch, and cell wall metabolism, were identified as important for the thickening process in the taproot of P. notoginseng. CONCLUSION: The results provide a molecular regulatory network of taproot thickening in P. notoginseng and facilitate the further characterization of the genes responsible for taproot formation in root medicinal plants or crops.


Assuntos
Redes Reguladoras de Genes , Metaboloma , Panax notoginseng/genética , Proteínas de Plantas/metabolismo , Transdução de Sinais , Transcriptoma , Regulação da Expressão Gênica de Plantas , Panax notoginseng/crescimento & desenvolvimento , Panax notoginseng/fisiologia , Proteínas de Plantas/genética , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/fisiologia
9.
Front Plant Sci ; 9: 1231, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30197651

RESUMO

Plant-derived terpenes are effective in treating chronic dysentery, rheumatism, hepatitis, and hyperlipemia. Thus, understanding the molecular basis of terpene biosynthesis in some terpene-abundant Chinese medicinal plants is of great importance. Abundant in mono- and sesqui-terpenes, Rhodomyrtus tomentosa (Ait.) Hassk, an evergreen shrub belonging to the family Myrtaceae, is widely used as a traditional Chinese medicine. In this study, (+)-α-pinene and ß-caryophyllene were detected to be the two major components in the leaves of R. tomentosa, in which (+)-α-pinene is higher in the young leaves than in the mature leaves, whereas the distribution of ß-caryophyllene is opposite. Genome-wide transcriptome analysis of leaves identified 138 unigenes potentially involved in terpenoid biosynthesis. By integrating known biosynthetic pathways for terpenoids, 7 candidate genes encoding terpene synthase (RtTPS1-7) that potentially catalyze the last step in pinene and caryophyllene biosynthesis were further characterized. Sequence alignment analysis showed that RtTPS1, RtTPS3 and RtTPS4 do not contain typical N-terminal transit peptides (62-64aa), thus probably producing multiple isomers and enantiomers by terpenoid isomerization. Further enzyme activity in vitro confirmed that RtTPS1-4 mainly produce (+)-α-pinene and (+)-ß-pinene, as well as small amounts of (-)-α-pinene and (-)-ß-pinene with GPP, while RtTPS1 and RtTPS3 are also active with FPP, producing ß-caryophyllene, along with a smaller amount of α-humulene. Our results deepen the understanding of molecular mechanisms of terpenes biosynthesis in Myrtaceae.

10.
J Clin Psychopharmacol ; 35(4): 406-10, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26066335

RESUMO

Major depressive disorder is a devastating mental illness leading to a lifetime prevalence of higher than 16% on individuals. The treatment delay and inevitable adverse effects are major limitations of current depression interventions. Emerging evidence indicates that curcumin produced significant antidepressant properties in depression in both rodents and humans without adverse effects. Therefore, it is necessary to further clarify the antidepressant actions of curcumin and the underlying mechanism in depressed patients. A total of 108 male adults aged between 31 and 59 years were systematically recruited in Tianjin Anding Hospital. Subjects were administered the Chinese version of 17-item Hamilton Depression Rating Scale and Montgomery-Asberg Depression Rating Scale that measures different scores of depressive symptoms. The subjects were asked to take 2 capsules containing either 1000 mg of curcumin or placebo soybean powder daily for 6 weeks on the basis of their current antidepressant medications. The plasma levels of interleukin 1ß, tumor necrosis factor α, brain-derived neurotrophic factor, and salivary cortisol were measured by enzyme-linked immunosorbent assay before and after curcumin or placebo treatment during the 6-week procedure. Chronic supplementation with curcumin produced significant antidepressant behavioral response in depressed patients by reduction of 17-item Hamilton Depression Rating Scale and Montgomery-Asberg Depression Rating Scale scores. Furthermore, curcumin decreases inflammatory cytokines interleukin 1ß and tumor necrosis factor α level, increases plasma brain-derived neurotrophic factor levels, and decreases salivary cortisol concentrations compared with placebo group. These findings indicate the potential benefits of further implications of supplementary administration of curcumin to reverse the development of depression and enhance the outcome of antidepressants treatment in major depressive disorder.


Assuntos
Antidepressivos/administração & dosagem , Citalopram/administração & dosagem , Curcumina/administração & dosagem , Transtorno Depressivo Maior/diagnóstico , Transtorno Depressivo Maior/tratamento farmacológico , Suplementos Nutricionais , Adulto , Fator Neurotrófico Derivado do Encéfalo/sangue , Transtorno Depressivo Maior/sangue , Método Duplo-Cego , Humanos , Hidrocortisona/antagonistas & inibidores , Hidrocortisona/sangue , Mediadores da Inflamação/antagonistas & inibidores , Mediadores da Inflamação/sangue , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Resultado do Tratamento
11.
PLoS One ; 7(9): e43973, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22957038

RESUMO

Posttraumatic stress disorder (PTSD) is characterized by acute and chronic changes in the stress response, manifested as conditioned fear memory. Previously formed memories that are susceptible to disruption immediately after retrieval undergo a protein synthesis-dependent process to become persistent, termed reconsolidation, a process that is regulated by many distinct molecular mechanisms that control gene expression. Increasing evidence supports the participation of the transcription factor NF-κB in the different phases of memory. Here, we demonstrate that inhibition of NF-κB in the basolateral amygdala (BLA), but not central nucleus of the amygdala, after memory reactivation impairs the retention of amygdala-dependent auditory fear conditioning (AFC). We used two independent pharmacological strategies to disrupt the reconsolidation of AFC. Bilateral intra-BLA infusion of sulfasalazine, an inhibitor of IκB kinase that activates NF-κB, and bilateral intra-BLA infusion of SN50, a direct inhibitor of the NF-κB DNA-binding complex, immediately after retrieval disrupted the reconsolidation of AFC. We also found that systemic pretreatment with sodium butyrate, a histone deacetylase inhibitor that enhances histone acetylation, in the amygdala rescued the disruption of reconsolidation induced by NF-κB inhibition in the BLA. These findings indicate that NF-κB activity in the BLA is required for memory reconsolidation in AFC, suggesting that NF-κB might be a potential pharmacotherapy target for posttraumatic stress disorder.


Assuntos
Tonsila do Cerebelo/metabolismo , Medo , Memória , NF-kappa B/metabolismo , Estimulação Acústica , Animais , Condicionamento Clássico , Modelos Animais de Doenças , Histonas/metabolismo , Quinase I-kappa B/metabolismo , Isobutiratos/farmacologia , Ratos , Ratos Sprague-Dawley , Transtornos de Estresse Pós-Traumáticos/diagnóstico , Transtornos de Estresse Pós-Traumáticos/fisiopatologia , Sulfassalazina/farmacologia
12.
J Integr Plant Biol ; 51(6): 574-80, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19522816

RESUMO

Aluminum-induced secretion of organic acids from the root apex has been demonstrated to be one major Al resistance mechanism in plants. However, whether the organic acid concentration is high enough to detoxify Al in the growth medium is frequently questioned. The genotypes of Al-resistant wheat, Cassia tora L. and buckwheat secrete malate, citrate and oxalate, respectively. In the present study we found that at a 35% inhibition of root elongation, the Al activities in the solution were 10, 20, and 50 muM with the corresponding malate, citrate, and oxalate exudation at the rates of 15, 20 and 21 nmol/cm(2) per 12 h, respectively, for the above three plant species. When exogenous organic acids were added to ameliorate Al toxicity, twofold and eightfold higher oxalate and malate concentrations were required to produce the equal effect by citrate. After the root apical cell walls were isolated and preincubated in 1 mM malate, oxalate or citrate solution overnight, the total amount of Al adsorbed to the cell walls all decreased significantly to a similar level, implying that these organic acids own an equal ability to protect the cell walls from binding Al. These findings suggest that protection of cell walls from binding Al by organic acids may contribute significantly to Al resistance.


Assuntos
Alumínio/metabolismo , Alumínio/farmacologia , Ácidos Carboxílicos/metabolismo , Parede Celular/efeitos dos fármacos , Parede Celular/metabolismo , Resistência a Medicamentos/efeitos dos fármacos , Magnoliopsida/efeitos dos fármacos , Adsorção/efeitos dos fármacos , Alumínio/toxicidade , Cassia/citologia , Cassia/efeitos dos fármacos , Cassia/metabolismo , Citratos/metabolismo , Fagopyrum/citologia , Fagopyrum/efeitos dos fármacos , Fagopyrum/metabolismo , Cinética , Magnoliopsida/citologia , Magnoliopsida/metabolismo , Malatos/metabolismo , Oxalatos/metabolismo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Triticum/citologia , Triticum/efeitos dos fármacos , Triticum/metabolismo
13.
J Chromatogr A ; 1167(2): 171-7, 2007 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-17854817

RESUMO

A systematic study was made on the identification and quantitative determination of polynuclear aromatic hydrocarbons (PAHs) in heavy products derived from coal and petroleum with high performance liquid chromatography (HPLC). After the separation of PAHs by a high-resolution column, identification was made through four methods: (1) the relative retention time (RRT) method, (2) the stop-flow-UV scanning method, (3) the method of fluorescence characteristic index Phi' and (4) the method of V' index at different UV wavelengths. For the quantitative determination of the components, methods of external standard (E-X), internal standard (I-N) and external standard-response factors (E-F) were compared. The E-F method was recommended by the present paper. For the determination of quantitative response factors (F) two methods were studied, including the HPLC peak-area method (Peak-area method) and the UV absorbance method using a UV spectrometer (absorbance method). The absorbance method was better and is recommended by the present paper. The F values of 30 PAHs from the two different methods are given. The samples analyzed include a coal tar pitch, a thermal cracking residue oil and a residual oil from ethylene production.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Carvão Mineral/análise , Petróleo/análise , Hidrocarbonetos Policíclicos Aromáticos/análise , Hidrocarbonetos Policíclicos Aromáticos/química , Reprodutibilidade dos Testes
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