Seasonal effect on phenolic content and antioxidant activity of young, mature and senescent leaves from Anredera cordifolia (Ten.) Steenis (Basellaceae)

Abstract Anredera cordifolia (Ten.) Steenis is a vine species native to Brazil that is considered an unconventional food plant and a medicinal species whose phenolic compounds exert antioxidant activity. Since the production of metabolites is determined by environmental factors and leaf maturity, it is important to track these changes in order to determine the best time to harvest. This study aimed to verify whether leaf phenology and seasonality cause variations in the amount of phenolic compounds and in the antioxidant activity of this species. The leaves were collected in different seasons between September 2018 and April 2019, and separated according to maturity: young, mature, and senescent. Daily atmospheric temperature and rainfall data were used to characterize the collection period. The total phenolic content (TPC), determined by Folin-Ciocalteu method, was significantly higher in the young leaves collected in winter, a season of lower temperatures. These leaves showed 54.4 mg of gallic acid equivalents per 100 g of dry matter (mg GAE 100 g-1DM). Other results averaged 25.6 mg GAE 100 g-1DM. The highest antioxidant activity, assessed via the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, regardless of leaf phenology, was observed in leaves harvested in autumn (73.7%) and winter (71.1%), seasons with lower rainfall. Leaves harvested in summer and spring had lower antioxidant action rates (54.3 e 37.5%, respectively). There was no significant correlation between the total phenolic content and antioxidant activity. Thus, the phenolic composition of A. cordifolia, and consequently its activity on free radicals, varies seasonally in response to temperature and rainfall, and may or may not interact with the age of the leaves.

Resumo Anredera cordifolia (Ten.) Steenis é uma trepadeira nativa do Brasil considerada uma planta alimentícia não convencional e uma espécie medicinal, cujos compostos fenólicos exercem ação antioxidante. Como a produção de metabólitos é determinada por fatores ambientais e pela maturidade das folhas, é importante acompanhar essas mudanças a fim de determinar a época de colheita. Neste estudo objetivou-se verificar se a fenologia foliar e a sazonalidade ocasionam variação na quantidade de compostos fenólicos e na ação antioxidante dessa espécie. As folhas foram coletadas em diferentes estações, entre setembro de 2018 e abril de 2019, e separadas conforme a maturidade: jovens, maduras e senescentes. Os dados diários de temperatura atmosférica e pluviosidade foram usados para caracterizar o período de coleta. O conteúdo de fenólicos totais (TPC), determinado pelo método Folin-Ciocalteu, foi significativamente maior nas folhas jovens coletadas no inverno, período caracterizado por temperaturas mais baixas. Estas folhas apresentaram 54,4 mg equivalentes de ácido gálico por 100 g de matéria seca (mg GAE 100 g-1DM) . A média dos outros resultados foi de 25,6 mg GAE 100 g-1DM. A maior ação antioxidante, avaliada pelo método do radical livre 2,2-difenil-1-picrilhidrazil (DPPH), foi observada em folhas colhidas no outono (73,7%) e inverno (71,1%), independentemente da fenologia foliar. Nestes períodos a pluviosidade foi menor. Folhas colhidas no verão e na primavera apresentaram menores percentuais de ação antioxidante (54,3 e 37,5%, respectivamente). Não houve correlação significativa entre o conteúdo de fenólicos totais e a atividade antioxidante. Conclui-se que a composição fenólica de A. cordifolia e, consequentemente, sua ação sobre radicais livres, varia sazonalmente em resposta à temperatura e pluviosidade, podendo ou não interagir com a idade das folhas.

Seasonal effect on phenolic content and antioxidant activity of young, mature and senescent leaves from Anredera cordifolia (Ten. ) Steenis (Basellaceae) / Efeito sazonal no conteúdo de fenólicos e ação antioxidante de folhas jovens, maduras e senescentes de Anredera cordifolia (Ten. ) Steenis (Basellaceae)

Anredera cordifolia (Ten.) Steenis is a vine species native to Brazil that is considered an unconventional food plant and a medicinal species whose phenolic compounds exert antioxidant activity. Since the production of metabolites is determined by environmental factors and leaf maturity, it is important to track these changes in order to determine the best time to harvest. This study aimed to verify whether leaf phenology and seasonality cause variations in the amount of phenolic compounds and in the antioxidant activity of this species. The leaves were collected in different seasons between September 2018 and April 2019, and separated according to maturity: young, mature, and senescent. Daily atmospheric temperature and rainfall data were used to characterize the collection period. The total phenolic content (TPC), determined by Folin-Ciocalteu method, was significantly higher in the young leaves collected in winter, a season of lower temperatures. These leaves showed 54.4 mg of gallic acid equivalents per 100 g of dry matter (mg GAE 100 g-1DM). Other results averaged 25.6 mg GAE 100 g-1DM. The highest antioxidant activity, assessed via the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method, regardless of leaf phenology, was observed in leaves harvested in autumn (73.7%) and winter (71.1%), seasons with lower rainfall. Leaves harvested in summer and spring had lower antioxidant action rates (54.3 e 37.5%, respectively). There was no significant correlation between the total phenolic content and antioxidant activity. Thus, the phenolic composition of A. cordifolia, and consequently its activity on free radicals, varies seasonally in response to temperature and rainfall, and may or may not interact with the age of the leaves.

Anredera cordifolia (Ten.) Steenis é uma trepadeira nativa do Brasil considerada uma planta alimentícia não convencional e uma espécie medicinal, cujos compostos fenólicos exercem ação antioxidante. Como a produção de metabólitos é determinada por fatores ambientais e pela maturidade das folhas, é importante acompanhar essas mudanças a fim de determinar a época de colheita. Neste estudo objetivou-se verificar se a fenologia foliar e a sazonalidade ocasionam variação na quantidade de compostos fenólicos e na ação antioxidante dessa espécie. As folhas foram coletadas em diferentes estações, entre setembro de 2018 e abril de 2019, e separadas conforme a maturidade: jovens, maduras e senescentes. Os dados diários de temperatura atmosférica e pluviosidade foram usados para caracterizar o período de coleta. O conteúdo de fenólicos totais (TPC), determinado pelo método Folin-Ciocalteu, foi significativamente maior nas folhas jovens coletadas no inverno, período caracterizado por temperaturas mais baixas. Estas folhas apresentaram 54,4 mg equivalentes de ácido gálico por 100 g de matéria seca (mg GAE 100 g-1DM) . A média dos outros resultados foi de 25,6 mg GAE 100 g-1DM. A maior ação antioxidante, avaliada pelo método do radical livre 2,2-difenil-1-picrilhidrazil (DPPH), foi observada em folhas colhidas no outono (73,7%) e inverno (71,1%), independentemente da fenologia foliar. Nestes períodos a pluviosidade foi menor. Folhas colhidas no verão e na primavera apresentaram menores percentuais de ação antioxidante (54,3 e 37,5%, respectivamente). Não houve correlação significativa entre o conteúdo de fenólicos totais e a atividade antioxidante. Conclui-se que a composição fenólica de A. cordifolia e, consequentemente, sua ação sobre radicais livres, varia sazonalmente em resposta à temperatura e pluviosidade, podendo ou não interagir com a idade das folhas.

Reserve mobilization and secondary metabolites during seed germination and seedling establishment of the tree Erythrina velutina (Fabaceae) / Movilización de reservas y metabolitos secundarios durante la germinación de semillas y el establecimiento de plántulas del árbol Erythrina velutina (Fabaceae)

Introduction: The lack of knowledge on seed germination and seedling establishment is a main constraint for the restoration of degraded areas, including the tropical dry forest known as Caatinga. Objective: To assess reserve and secondary metabolite mobilization during seed germination and seedling establishment in Erythina velutina. Methods: We scarified, disinfected, imbibed, sown between towel paper, and incubated seeds under controlled conditions. We hydroponically cultivated seedlings in a greenhouse. We harvested cotyledons at seed imbibition, radicle protrusion, hypocotyl emergence, apical hook formation and expansion of cordiform leaves, first trifoliate leaf, and second trifoliate leaf. Results: Seeds contained approximately 20 % starch, 14.5 % storage proteins, 11.6 % neutral lipids, and 5.7 % non-reducing sugars on a dry weight basis. Soluble sugars were mainly consumed from hypocotyl emergence to apical hook formation, while major reserves were mobilized from apical hook formation to expansion of first trifoliate leaf. Enzymatic activity increased from mid to late seedling establishment, causing the mobilization of starch, oils, and proteins. Terpenoid-derivatives, flavonoids, phenolic acids, and alkaloids were detected. Flavonoids and phenolic acids were present at almost all stages and terpenoid-derivatives disappeared at expansion of cordiform leaves. Conclusion: Soluble sugars support early seedling growth, while starch, oils and proteins are simultaneously mobilized from mid to late establishment by amylases, lipases, and acid proteases. The cotyledons contain secondary metabolites, which may act in seedling defense. High content of reserves and presence of secondary metabolites in the cotyledons could enable E. velutina seedlings endure stress, validating their use in the restoration of degraded areas.

Introducción: La falta de conocimiento sobre la germinación de semillas y el establecimiento de plántulas es una de las principales limitaciones para la restauración de áreas degradadas, incluido el bosque seco tropical conocido como Caatinga. Objetivo: Evaluar la movilización de reservas y metabolitos secundarios durante estas etapas de desarrollo en Erythina velutina. Métodos: Las semillas fueron escarificadas, desinfectadas, embebidas, sembradas entre toallas de papel e incubadas bajo condiciones controladas. Cultivamos las plántulas hidropónicamente en un invernadero. Recolectamos los cotiledones en la imbibición de la semilla, la protrusión de la radícula, la emergencia del hipocótilo, la formación del gancho apical y la expansión de las hojas cordiformes, la primera y segunda hoja trifoliada. Resultados: Las semillas contenían 20 % de almidón, 14.5 % de proteínas de almacenamiento, 11.6 % de lípidos neutros y 5.7 % de azúcares no reductores en peso seco. Los azúcares solubles se consumieron desde la emergencia del hipocótilo hasta la formación del gancho apical. Las principales reservas se movilizaron desde la formación del gancho apical hasta la expansión de la primera hoja trifoliada. La actividad enzimática aumentó desde la mitad hasta el final del establecimiento de las plántulas, movilizando almidón, aceites y proteínas. Se detectaron derivados de terpenoides, flavonoides, ácidos fenólicos y alcaloides. Los flavonoides y los ácidos fenólicos estuvieron en casi todas las etapas y los derivados terpenoides desaparecieron en la expansión de las hojas cordiformes. Conclusión: Los azúcares solubles apoyan el crecimiento temprano de las plántulas; el almidón, los aceites y las proteínas se movilizan simultáneamente desde el establecimiento medio hasta el final por amilasas, lipasas y proteasas ácidas. Los cotiledones contienen metabolitos secundarios, que pueden actuar en la defensa de las plántulas. El alto contenido de reservas y los metabolitos secundarios en los cotiledones podría permitir que las plántulas de E. velutina toleren estrés, validando su uso en la restauración de áreas degradadas.

Trichoderma spp., una alternativa para la agricultura sostenible: una revisión

En la actualidad uno de los retos a los que se enfrentan los agricultores es producir alimentos bajo las inclemencias climáticas. Para el 2050, se estima un aumento en la demanda en producción de alimentos básicos a causa del incremento demográfico, aumentando con ello el detrimento de los recursos naturales. Dentro de las alternativas biológicas está el uso de insumos a base de microorganismos benéficos, como el género Trichoderma. Los cuales se han utilizado en los campos agrícolas para el control biológico contra un gran número de fitopatógenos. Sin embargo, aún son poco conocidas otras propiedades benéficas de este género para las plantas que coloniza y el ecosistema. Se realizó una búsqueda de artículos científicos en Academic Search Ultimate, BioOne, Acsess, Esmerald, Fuente Académica, ScienceDirect y Springer, entre 2015 y 2023, con dos excepciones de años anteriores. Se utilizó la palabra clave "Trichoderma" y aquellas relacionadas con interacciones microbianas y su aplicación agrícola. Esta revisión resume los hallazgos bibliográficos actuales de este género que muestran su alta capacidad hacia el desarrollo sostenible de los agroecosistemas. Varias investigaciones reportan su capacidad de inducir la defensa vegetal, la promoción del crecimiento y desarrollo radicular, así como la estimulación y síntesis de sustancias que contribuyen a fortalecer la fertilidad del suelo. Con ello mejora los rendimientos de los cultivos a los que se encuentra asociado. En definitiva, la aplicación de Trichoderma puede coadyuvar a disminuir los efectos negativos ocasionados por el uso de agroquímicos y fertilizantes sintéticos, contribuyendo a una producción más sostenible.

Currently, one of the most critical challenges facing farmers is the production of food under adverse weather conditions. By 2050, an increase in the production of staple foods is estimated due to demographics, thereby increasing the depletion of natural resources. Among the biological alternatives is the use of inputs based on beneficial microorganisms such as the Trichoderma genus, which have been used in agricultural fields for biological control against a large number of phytopathogens. However, other beneficial properties of this genus for the plants it colonizes, and the ecosystem are still little known. Therefore, a search for scientific articles was carried out in Academic Search Ultimate, BioOne, Acsess, Esmerald, Fuente Academic, ScienceDirect and Springer, between 2015 and 2023, with two exceptions from previous years. The keyword "Trichoderma" was used and those related to microbial interactions and their agricultural application. Therefore, this review summarizes the current bibliographic findings of this genus, that shows its high capacity towards the sustainable development of agroecosystems. Several investigations report its ability to induce plant defense, promote growth and root development, and stimulate and synthesize substances that help strengthen soil fertility. This improves the yields of the crops to which they are associated. With this, the application of Trichoderma can reduce the negative effects caused by the use of agrochemicals and synthetic fertilizers, contributing to a more sustainable production.

A comparative study on surgical outcomes of trabeculectomy with and without anti-metabolites in juvenile open-angle glaucoma

Purpose: To compare the surgical outcomes of trabeculectomy with and without anti?metabolites in patients with juvenile open?angle glaucoma (JOAG). Methods: This retrospective comparative case series included 98 eyes of 66 patients with JOAG who underwent either trabeculectomy without anti?metabolites (group A, n = 53 eyes) or with anti?metabolites (group B, n = 45 eyes) with a minimum of 2 years follow?up. The main outcome measures were intra?ocular pressure (IOP), number of glaucoma medications, visual acuity, additional surgical interventions, surgical complications, and risk factors for failure. Surgical failure was defined as IOP >18 mmHg or failure to reduce IOP by <30% from the baseline value or IOP ?5 mmHg or re?operation for refractory glaucoma or a complication or loss of light perception vision. Results: The mean post?operative IOP reduced significantly from baseline at all post?operative visits until 6 months and thereafter. The cumulative probability of failure at 2 years was 28.7% in group A [95% confidence interval (CI) = 17.6–44.8%] and 29.1% in group B (95% CI = 17.1–46.7%) (P = 0.78). Surgical complications occurred in 18 eyes (34%) in group A and 19 eyes (42%) in group B. Re?operations for glaucoma or complications were performed in two eyes (3.8%) in group A and two eyes (4.4%) in group B. Cox?hazard regression model revealed male gender (HR = 0.29; P = 0.008), baseline high IOP (HR = 0.95; P = 0.002), and an increased number of pre?operative glaucoma medications (HR = 2.08; P = 0.010) as significant factors associated with failure. Conclusion: Our study results on trabeculectomy in JOAG revealed a success of 71% in both groups at 2 years follow?up. There was no significant difference in success or failure rates between the two groups. The risk factors for poor surgical outcome in JOAG were male gender, baseline high IOP, and an increased number of glaucoma medications

Phytochemical Characterization and Systemic Exposure of Bioactive Metabolites of the Aqueous Extracts of Ficus thonningii Blume (Moraceae) on Albino Wistar Rats

Ficus thonningii (Blume) is considered as a herbal plant with well documented biological activity in the management of several diseases in the tropics. However, there is a gap of information on its safety and proof of efficacy in evidence-based medicine. The objective of this study was to characterize the bioactive metabolites of the hydro-ethanolic extract of the stem bark of Ficus. thonningii and in vivo evaluation of the systemic exposure of the bioactive metabolite. Phytochemical screening was done using standard extraction techniques, and test according to methods adopted from Sofowora and collaborators. Quantitative analysis was done using spectrophotometer of plant extract with different reference standards. Analysis of the animals' plasma following administration of the extract was used to investigate systemic exposure to confirmed the presence of absence of metabolites in systemic circulation. This work shows that F. thonningii (Blume) stem bark hydro-ethanolic extract contains polyphenols, saponins, alkaloids, flavonoids, catechic tannins, gallic tannins, coumarins, quinones, phlobatannins. This study shows that the hydro-ethanolic extract of F. thonningii contains total phenolic content of 192,27 ± 3,40 mgEQ/MS g gallic acid and total flavonoid content of 103,59 ± 15,72 mgEQ/MS quercetin. This study shows that the secondary metabolites in the hydro-ethanolic extract of the stem bark of F. thonningii (Blume) were not detected in plasma and not bioavailable.

Potencial antiinflamatorio del plátano Musa spp. ABB para la elaboración de fitofármacos

Los plátanos pertenecientes al género Musa spp. ABB han sido utilizados para el tratamiento de la inflamación y el dolor. La búsqueda de antiinflamatorios de origen natural, más que una alternativa, constituye una terapia complementaria del tratamiento de la inflamación, ya que los antiinflamatorios no esteroideos son los más utilizados para enfermedades inflamatorias agudas o crónicas, pero a la vez presentan efectos secundarios gastrointestinales, cardiovasculares y renales, por lo que no es recomendable su uso prolongado. En este trabajo se ofrece información actualizada sobre los efectos farmacológicos y los metabolitos secundarios del grupo Musa spp. ABB, para su uso en la elaboración de un medicamento antiinflamatorio. Los procedimientos realizados incluyen el análisis documental y bibliográfico -en bases de datos- de artículos científicos, tesis doctorales y comunicaciones científicas; el registro de la información, y la elaboración del artículo científico, teniendo en cuenta la actualización de la información y los sitios donde se promueve la misma. Se han comprobado, a partir de extractos acuosos y alcohólicos de la planta, efectos farmacológicos tales como antiinflamatorio, antioxidante, analgésico, gastroprotector, inmunomodulador, antidiabético, anticancerígeno, antibacteriano, antiviral, hipolipemiante, antidiarreico, antihipertensivo, hepatoprotector, cicatrizante, antiulceroso y antifúngico. Los efectos antiinflamatorios y antioxidantes están relacionados con la presencia de fenoles, ácidos fenólicos, flavonoides, fitoesteroles y carotenos. Es muy limitado el empleo de extractos del grupo Musa spp. ABB en medicamentos, de ahí la propuesta de desarrollar una crema antiinflamatoria como alternativa o complemento para el tratamiento de múltiples enfermedades inflamatorias.

Plantains belonging to the genus Musa spp. ABB have been used for the treatment of infection and pain. The search for anti-inflammatories of natural origins, more than an alternative, is a complementary therapy for the treatment of inflammation, since non-steroidal anti-inflammatories are the most used for acute or chronic inflammatory diseases, but at the same time they present gastrointestinal, cardiovascular and renal side effects, so their prolonged use is not recommended. In this paper, updated information is provided on the pharmacological effects and secondary metabolites of the Musa ssp. ABB group, for its use in elaborating an anti-inflammatory drug. The procedures carried out include documental and bibliographic analysis -in databases-of scientific articles, doctoral theses and scientific communications; the information record and preparation of the article taking into account the information updating and the sites where it is supported. From aqueous and alcoholic extracts of the plant, the following effects were proven: anti-inflammatory, antioxidant, analgesic, gastro-protective, inmuno-modulator, antidiabetic, anticancer, antibacterial, antiviral, lipid-lowering, antidiarrheal, antihypertensive, hepatoprotective, healing, anti-ulcerative and antifungal. The anti-inflammatory and antioxidant effects are related to the presence of phenols, phenolic acids, flavonoids, phytosterols and carotenes. The use of Musa spp. ABB group in drugs is very limited, therefore the proposal of developing an anti-inflammatory cream to treat multiple inflammatory diseases.

Atividade in vitro de extratos e frações de Copernicia prunifera (Arecaceae) contra bactérias de importância clínica em saúde pública / In vitro activity of extracts and fractions of Copernicia prunifera (Arecaceae) against bacteria of clinical importance in public health / Actividad in vitro de extractos y fracciones de Copernicia prunifera (Arecaceae) frente a bacterias de importancia clínica en salud pública

Introdução: O aumento contínuo da resistência bacteriana aos antibióticos convencionais é um problema de importância global. Encontrar produtos como alternativas terapêuticas naturais é essencial. As plantas medicinais possuem uma composição química muito rica, que podem ser estruturalmente otimizadas e processadas em novos antimicrobianos. Objetivo: Avaliar o potencial antibacteriano frente a microrganismos humanos potencialmente patogênicos do extrato etanólico e frações de Copernicia prunifera. Metodologia: A triagem fitoquímica de plantas foi realizada usando métodos de precipitação e coloração e a atividade antibacteriana utilizando o método de difusão em disco e microdiluição em caldo contra cepas padronizadas de Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa e Staphylococcus aureus. Resultados: A triagem fitoquímica revela a presença de taninos, flavonoides, esteroides, triterpernóides, saponinas e alcaloides. Os extratos etanólico e frações da casca do caule e folhas tiveram atividade inibitória contra S. aureus e K. pneumonie com zona de inibição que variou de 7,0±1,73 a 9,33±0,58 mm pelo método de difusão em disco. Pelo método de microdiluição em caldo os extratos foram satisfatórios somente contra K. pneumoniae (CIM = 125 a 1000 µg/mL) S. aureus, P. aeruginosa e E. coli se mostraram resistentes aos testes (CIM > 1000 µg/mL). Conclusão: Esses resultados fornecem uma base para futuras investigações em modelos in vivo, para que os compostos de C. prunifera possam ser aplicados no desenvolvimento de novos agentes antimicrobianos contra K. pneumoniae.

Introduction: The continuous increase in bacterial resistance to conventional antibiotics is a problem of global importance. Finding products as natural therapeutic alternatives is essential. Medicinal plants have a very rich chemical composition, which can be structurally optimized and processed into novel antimicrobials. Objective: To evaluate the antibacterial potential against potentially pathogenic human microorganisms of the ethanolic extract and fractions of Copernicia prunifera. Methodology: Phytochemical screening of plants was performed using precipitation and staining methods and antibacterial activity using the disk diffusion and broth microdilution method against standardized strains of Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus. Results: Phytochemical screening reveals the presence of tannins, flavonoids, steroids, triterpernoids, saponins and alkaloids. The ethanolic extracts and fractions of stem bark and leaves had inhibitory activity against S. aureus and K. pneumonie with zone of inhibition ranging from 7.0±1.73 to 9.33±0.58 mm by disc diffusion method. By broth microdilution method the extracts were satisfactory only against K. pneumoniae (MIC = 125 to 1000 µg/mL) S. aureus, P. aeruginosa and E. coli were resistant to the tests (MIC > 1000 µg/mL). Conclusion: These results provide a basis for further investigation in in vivo models, so that compounds from C. prunifera can be applied in the development of new antimicrobial agents against K. pneumoniae.

Introducción: El continuo aumento de la resistencia bacteriana a los antibióticos convencionales es un problema de importancia mundial. Es esencial encontrar productos como alternativas terapéuticas naturales. Las plantas medicinales tienen una composición química muy rica, que puede optimizarse estructuralmente y transformarse en nuevos antimicrobianos. Objetivo: Evaluar el potencial antibacteriano frente a microorganismos humanos potencialmente patógenos del extracto etanólico y fracciones de Copernicia prunifera. Metodología: Se realizó el cribado fitoquímico de las plantas mediante los métodos de precipitación y tinción y la actividad antibacteriana mediante el método de difusión en disco y microdilución en caldo frente a cepas estandarizadas de Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa y Staphylococcus aureus. Resultados: El cribado fitoquímico revela la presencia de taninos, flavonoides, esteroides, triterpernoides, saponinas y alcaloides. Los extractos etanólicos y las fracciones de la corteza del tallo y las hojas presentaron actividad inhibitoria contra S. aureus y K. pneumonie con una zona de inhibición que osciló entre 7,0±1,73 y 9,33±0,58 mm por el método de difusión en disco. Por el método de microdilución en caldo, los extractos sólo fueron satisfactorios frente a K. pneumoniae (CMI = 125 a 1000 µg/mL). S. aureus, P. aeruginosa y E. coli fueron resistentes a las pruebas (CMI > 1000 µg/mL). Conclusiones: Estos resultados proporcionan una base para futuras investigaciones en modelos in vivo, de modo que los compuestos de C. prunifera puedan aplicarse en el desarrollo de nuevos agentes antimicrobianos contra K. pneumoniae.

Detection of Carbamazepine and Its Metabolites in Blood Samples by LC-MS/MS / 法医学杂志

OBJECTIVES@#To establish a method for the detection of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples by liquid chromatography-tandem mass spectrometry (LC-MS/MS).@*METHODS@#The blood samples were treated with 1-butyl-3-methylimidazolium hexafluorophosphate as an extraction solvent. The samples were extracted by ultrasound-assisted extraction and separated by ZORBAX Eclipse Plus C18, 95Å column. The mobile phase A aqueous solution containing 0.1% formic acid and 10 mmol/L ammonium acetate, and mobile phase B mixed organic solvent containing acetonitrile/methanol (Vacetonitrile∶Vmethanol=2∶3) were used for gradient elution at the flow rate of 1.00 mL/min. An electrospray ion source in positive mode was used for detection in the multiple reaction monitoring.@*RESULTS@#The linearities of carbamazepine and its metabolites 10,11-dihydro-10,11-epoxycarbamazepine and 10,11-dihydro-10-hydroxycarbamazepine in blood samples were good within the corresponding range, with correlation coefficients (r) greater than 0.995 6. The limits of detection were 3.00, 0.40 and 1.30 ng/mL, respectively. The limit of quantitation were 8.00, 1.00 and 5.00 ng/mL, respectively. The extraction recoveries ranged from 76.00% to 106.44%. The relative standard deviations of the intra-day and inter-day precisions were less than 16%. Carbamazepine and its main metabolite 10,11-dihydro-10,11-epoxycarbamazepine were detected in blood samples of death cases with a mass concentration of 2.71 μg/mL and 252.14 ng/mL, respectively.@*CONCLUSIONS@#This method has high sensitivity and good selectivity, which is suitable for the detection of carbamazepine and its metabolites in blood samples, and can be used for carbamazepine-related forensic identifications.

The correlation between intestinal flora and glucose metabolism during pregnancy and the research progress on the application of probiotics / 中华临床营养杂志

Gut microbiota is the microbial community that resides on the surface of human intestinal mucosa. During normal pregnancy, the composition of gut microbiota may change dynamically with the progress of pregnancy. Gestational diabetes mellitus (GDM) is a common complication of pregnancy, which can affect maternal and neonatal intestinal flora, and affect the long-term glucose metabolism of mothers and infants through exacerbating insulin resistance and promoting inflammatory response. Adjustment of dietary structure and application of probiotics may regulate intestinal microbiota and improve maternal and neonatal glucose metabolism in GDM. Here we reviewed the correlation between intestinal flora and glucose metabolism during pregnancy, and discussed the effects of diet and probiotics on gut microbiota.

Targeted metabolomic analysis of phenylpropanoid metabolites in Astragalus membranaceus var. mongholicus in response to Fusarium solani infection / 药学学报

Root rot severely restricts the sustainable development of Astragalus membranaceus var. mongholicus (AMM) industry. Resistance breeding is an economical and environmentally safe way to manage the disease and its key lies in the obtaining of resistance indicators. This study aimed to quickly and accurately screen the resistance-related (RR) metabolites so as to provide reference for the screening of indicators of AMM breeding for resistance. LC-MS-based targeted metabolomics and real-time quantitative PCR technology were employed, in combination with multivariate statistical analysis, in analyzing the dynamic changes of phenylpropanoid metabolites in AMM in response to root rot pathogen Fusarium solani (FS) infection and identifying the differential metabolites. The LC-MS method established showed high sensitivity; each metabolite had a good linear relationship (R2 ≥ 0.968 9) in the corresponding linear range of the respective standard curve; the recoveries and the relative standard deviations (RSDs) (n = 6) ranged from 70% to 107% and from 1.2% to 9.9%, respectively. Obvious disturbances were observed in the changes of the targeted metabolites in AMM infected by FS. These metabolites, compared with the mock-inoculated (CK) group, showed different up or down regulation with time series. Calycosin-7-O-β-D-glucoside, ononin, calycosin and formononetin were identified as differential metabolites, and they all belong to flavonoids. The first three compounds were significantly negatively correlated (r ≤ -0.97, P < 0.05) with the content of FS in the root of AMM. As potential RR metabolites, they are helpful in obtaining promising resistance indicators for AMM against FS infection.

Secondary metabolites of endophytic fungus Aspergillus sp. J218 from Anectochilus roxburhii / 药学实践杂志

Objective To isolate the secondary metabolites of endophytic fungus Aspergillus sp. J218 from Anectochilus roxburhii. Methods Different chromatographic methods, including Sephadex LH-20 and silica gel chromatography as well as HPLC, were used to isolate compounds from the EtOAc fraction of the solid fermentation of J218, and their structures were identified by spectral methods. Results Ten compounds were isolated from the fermentation of J218 and their structures were individually identified as kotanin（1）, flavasperone（2）, aurasperone B（3）, fonsecinone B（4）, fonsecinone D（5）, ensidol A（6）, fonsecinone A（7）, fonsecinone C（8）, aurasperone A（9）, and fonsecinone F（10）. Conclusion Most compounds isolated from endophytic fungus Aspergillus sp. J218 in Anectochilus roxburhii were identified as dimeric naphthopyrones. These results suggest that this strain contains rich dimerization synthase, which could provide clues for the further exploration of the rational biosynthesis pathway of dimeric naphthopyrones in this strain.

Biological Mechanism of Drought Improving Quality of Rhizoma Atractylodis Chinensis / 中国实验方剂学杂志

ObjectiveTo explore the biological mechanism of drought improving the quality of Rhizoma Atractylodis Chinensis and establish a new method for the production of high-quality medicinal materials. MethodThe fresh roots of Atractylodes chinensis were soaked in 0 （control）， 5%， 10%， and 20% PEG-6000 solutions. The changes in reactive oxygen species （ROS） level， antioxidant enzyme activity， activities of key enzymes in primary metabolism and secondary metabolism， and content of secondary metabolites were compared. ResultCompared with the control group， the treatment with 20% PEG for 2 days elevated the levels of superoxide anion radicals （O2-·）， hydrogen peroxide （H2O2）， and malondialdehyde （MDA） by 172.5%， 56.9%， and 14.7%， respectively. The treatment did not change the activity of superoxide dismutase （SOD）， reduced the peroxidase （POD） activity， and increased the catalase （CAT） activity by 10.8%. It increased the activities of 3-hydroxy-3-methylglutaryl-CoA reductase （HMGR）， phosphoenolpyruvate carboxylase （PEPC）， and acetyl-CoA carboxylase （ACC） by 49.9%， 12.1%， and 19.0%， respectively. Furthermore， the content of atractylodin， β-eudesmol， atractylone， and atractylenolide Ⅱ was increased by 51.0%， 36.9%， 47.1%， and 91.5%， respectively. The simulated drought stress can cause the burst of ROS in the fresh roots of A. chinensis， induce the physiological state of plants under drought， change the antioxidant system， and promote the massive synthesis of secondary metabolites in a short time. ConclusionPEG-6000-simulated drought stress can greatly improve the quality of A. chinensis in cultivation.

Qualitative Analysis of Metabolites of Aristolochiae Fructus Aqueous Extract in Rats / 中国实验方剂学杂志

ObjectiveBased on ultra performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MSE） technique， we identified qualitatively the metabolites of aristolochic acid（AAs） in rat in order to analyze the metabolic differences between water extract of Aristolochiae fructus（AFE） and Aristolochic acid Ⅰ（AAⅠ）. MethodSD rats were selected and administered AFE（110 g·kg-1·d-1） or AAⅠ（5 mg·kg-1·d-1） by oral for 5 days， respectively. Serum， urine and feces were collected after administration. Through sample pretreatment， ACQUITY UPLC BEH C18 column（2.1 mm×100 mm， 1.7 μm） was used with the mobile phase of 0.01% formic acid methanol（A）-0.01% formic acid water（B， containing 5 mmol·L-1 ammonium acetate） for gradient elution（0-1 min， 10%B； 1-7 min， 10%-75%B； 7-7.2 min， 75%-95%B； 7.2-10.2 min， 95%B； 10.2-10.3 min， 95%-10%B； 10.3-12 min， 10%B） at a flow rate of 0.3 mL·min-1. Positive ion mode of electrospray ionization（ESI+） was performed in the scanning range of m/z 100-1 200. In combination with UNIFI 1.9.4.053 system， the Pathway-MSE was used to qualitatively analyze and identify the AAs prototype and related metabolites in biological samples（serum， urine and feces）， and to compare the similarities and differences of metabolites in rats in the subacute toxicity test between AFE group and AAⅠ group. ResultCompared with AAⅠ group， 6， 10， 13 common metabolites and 14， 20， 30 unique metabolites were identified in biological samples（serum， urine and feces） of AFE group， respectively. Moreover， the main AAs components always followed the metabolic processes of demethylation， nitrate reduction and conjugation. Compared with common metabolites in AAⅠ group， prototype components of AAⅠ in serum and most metabolic derivatives of AAⅠ［AAⅠa， aristolochic lactam Ⅰ（ALⅠ）a， 7-OHALⅠ and its conjugated derivatives］ in biological samples were significantly increased in AFE group（P<0.05， P<0.01）， except that the metabolic amount of ALⅠ in feces of AFE group was remarkably lowed than that of AAⅠ group（P<0.01）. In addition， a variety of special ALⅠ efflux derivatives were also identified in the urine and feces of the AFE group. ConclusionAlthough major AAs components in AFE all show similar metabolic rules as AAⅠ components in vivo， the coexistence of multiple AAs components in Aristolochiae Fructus may affect the metabolism of AAⅠ， and achieve the attenuating effect by increasing the metabolic effection of AAⅠ and ALⅠ.

Characterization and application of exposure biomarkers of polycyclic aromatic hydrocarbons in diesel exhaust / 环境与职业医学

Diesel exhaust (DE), Group 1 carcinogen, is an important source of air pollutants. Studies show that DE exposure associates with elevated incidences of respiratory and cardiovascular diseases. The toxic effects of DE are closely related to its components. Polycyclic aromatic hydrocarbons (PAHs) are one of the main toxic components in DE and are often used as human exposure biomarkers to DE. However, the exposure assessment of DE using PAHs as biomarkers could be interfered due to the other sources of PAHs. Therefore, identification of highly specific and reliable PAHs sourced biomarkers of DE exposure has become a hotspot of current research. New biomarkers of DE may play an important role in determining human exposure to DE and establishing dose-response relationship of DE exposure and health outcomes of interest. This paper focused on current progress in terms of PAHs sourced biomarkers of human exposure to DE with the following aims: (1) to clarify the types of PAHs sourced biomarkers to DE; (2) to explore the applicability and limitations of PAHs sourced biomarkers for DE exposure assessment in occupational exposure and environmental exposure analysis; and (3) to summarize the analysis methods for PAHs sourced exposure biomarkers in human urine samples and compare the advantages and disadvantages of different analytical methods.

A systematic review of metabolomic studies on the mechanism of myopia / 中华实验眼科杂志

Myopia has become a global public health concern with its increasing prevalence.It is the interaction result of genetic and environmental factors.Exploration of the changes of metabolites in myopia is helpful to know new clues about its pathogenic mechanism.Metabolomics focuses on the integral analysis of all small molecular metabolites (relative molecular mass <1 000) which form a biological system and it is used as an effective tool to discover potential biomarkers.Metabolomic analysis of the myopic population could discover the metabolic changes related to myopia and screen the markers with potential biological significance, which can be used in the early diagnosis and treatment of myopia.It has been found that metabolites related to oxidative stress and inflammation play an important role in the development of myopia.Abnormal energy metabolism and amino acid metabolism are associated with myopic fundus changes.In addition, classical myopia-associated metabolites such as retinoic acid, dopamine and vitamin D, other metabolites such as melatonin, cyclic adenosine monophosphate and 5-hydroxy indole acetic acid, as well as multiple metabolic pathways such as fatty acid metabolism and mitochondrial metabolism are all closely related to myopia.This article systematically reviewed metabolomics researches on myopia, providing clues for better prevention and control of myopia in the future.

Advances in intestinal flora metabolites deoxycholic acid and digestive diseases in children / 国际儿科学杂志

It is the hotspot that studying the interplay and mechanism between intestinal flora metabolites and diseases.Deoxycholic acid, one of the intestinal flora metabolites, is one of the most abundant secondary bile acids in human intestinal tract, which is corelated with many diseases, while the mechanisms remain unclear.The imbalance of deoxycholic acid is connected with the intestinal flora disorder and high fat diet, which could result in several immunoreaction and inflammatory reaction.In this review, the interaction between deoxycholic acid and digestive diseases in children, such as non-alcoholic fatty liver disease, inflammatory bowel disease and irritable bowel syndrome, is discussed to explore their related mechanism, so as to clarify the direction of further study on the influence of intestinal microbiota metabolites deoxycholic acid on the human body.

Influences of intestinal flora metabolites on neonatal diseases / 国际儿科学杂志

The intestinal tract is an essential digestive organ of the human body, which is known as the "second brain". The intestinal flora disorder is closely related to the occurrence of host diseases.It has been found that the dysbiosis of intestinal flora plays an important role in the development of neonatal diseases.Gut microflora metabolites are bioactive, and the key metabolites can regulate or affect the host′s metabolic changes through different metabolic pathways.The metabolites of the neonatal intestine participate in and affect the progression and outcome of the diseases, and determine their short- and long-term quality of life.This review summarizes the effects of gut flora metabolites on neonatal diseases.

Method establishment and clinical practice for concentration determination of caffeine and its metabolites in urine / 中国药房

OBJECTIVE To establish a method for concentration determination of caffeine and its three metabolites， theophylline， paraxanthine and theobromine in urine， and apply it in clinical practice. METHODS Using caffeine-13C3-d3 as internal standard （IS）， and the urine samples were protein precipitated with acetonitrile； HPLC-MS/MS method was adopted to determine the concentrations of caffeine and its three metabolites. The determination was performed on Waters ACQUITY UPLC® BEH HILIC column with mobile phase consisting of 60 mmol/L ammonium acetate （A）-acetonitrile （B） （gradient elution） at the flow rate of 0.5 mL/min. The column temperature was set at 38 ℃ ， and the sample size was 2 μL. The electrospray ionization detection was operated in a positive mode by multiple reaction monitoring. The detection ions for quantitative analysis were m/z 195.1→110.0 for caffeine， m/z 181.1→124.0 for theophylline， m/z 181.1→124.0 for paraxanthine， m/z 181.1→138.0 for theobromine， and m/z 198.1→ 140.1 for IS. The above method was used to determine the concentrations of caffeine and its three metabolites in the urine of 19 infants with apnea of prematurity （AOP）. RESULTS The linear ranges of mass concentration of caffeine， theophylline， paraxanthin and theobromine were 0.200-200， 0.050-50.0，0.050 0-50.0， and 0.100-100 μg/mL， respectively. The lower limits of quantification were 0.200， 0.050， 0.050 and 0.100 μg/mL （r＞0.990）， respectively. RSDs of intra-day and intra- day precision were not above 10.37%， and matrix factors were 85.68%-109.90%； extraction recoveries were 93.53%-109.40% （RSD≤15%）， and RSDs of stability tests were all lower than 15%. The concentrations of caffeine and its three metabolites in the urine of 19 cases were （27.346±7.951）， （0.351±0.223）， （0.428±0.395） and （0.472±0.374） μg/mL， respectively. CONCLUSIONS The established HPLC-MS/MS method is simple， sensitive and can be used for the determination of caffeine and its three metabolites in urine samples of AOP.

Analysis on Metabolites and Metabolic Pathways of Limonin in Rats by UHPLC-Q-Orbitrap HRMS / 中国实验方剂学杂志

ObjectiveUltra-high performance liquid chromatography-quadrupole-electrostatic field orbitrap high resolution mass spectrometry（UHPLC-Q-Orbitrap HRMS） was used to identify the metabolites of limonin in rats， and to explore the gender differences in the distribution of prototype components and metabolites in rats after single dose intragastric administration of limonin， as well as to speculate the metabolic pathways. MethodThe separation was performed on a Thermo Scientific Accucore™ C18 column（3 mm×100 mm， 2.6 μm） with 0.1% formic acid aqueous solution（A）-0.1% formic acid acetonitrile solution（B） as mobile phase in a gradient elution mode（0-1 min， 5%B； 1-6 min， 5%-20%B； 6-18 min， 20%-50%B； 18-23 min， 50%-80%B； 23-25 min， 80%-95%B； 25-30 min， 95%B） at a flow rate of 0.3 mL·min-1 and a column temperature of 30 ℃. MS data of biological samples were collected under the positive ion mode of electrospray ionization（ESI） and in the scanning range of m/z 100-1 500. Plasma， tissues（heart， liver， spleen， lung， kidney， stomach and small intestine）， urine and fecal samples were collected and prepared after intragastric administration， and the prototype component and metabolites of limonin were identified. ResultThe prototype component of limonin were detected in the feces， stomach， small intestine of female and male rats， and in the heart， liver， spleen， lung and kidney tissues of female rats. A total of 23 metabolites related to limonin were detected in rats， of which 2， 1， 5， 4， 23 metabolites were detected in liver， stomach， small intestine， urine and feces， respectively， and the main metabolic pathways were hydrolysis， reduction， hydroxylation and methylation， etc. The distribution of some metabolites differed between male and female rats. ConclusionThe prototype component of limonin are mainly distributed in the stomach and small intestine in rats， and the distribution of prototype component and some metabolites are different by gender. Limonin is mainly excreted through feces with phase Ⅰ metabolites as the main ones. The results of this study can provide a reference for further elucidation of the effect of gender differences on the metabolism of limonin in vivo and its mechanism of action.