Computational Evaluation of Bioactive Compounds from Dysphania ambrosioides Leaves.

Dysphania ambrosioides has been reported to have many medicinal properties, due to its possession of a multitude of biologically active molecules contained in its leaves. However, very few studies have been reported to evaluate their pharmacological properties. Consequently, in the present study, many computational tools have been performed to predict drug similarity and ADMET properties. Besides, the inhibitory potential of D.ambrosioides major compounds against Bacterial, Fungal and cardiovascular main receptor targets has been investigated. This study suggests that Carvone oxide, 5-Isopropenyl-2-Methylenecyclohexanol, and Caryophyllene oxide were the most active molecules belonging to D. ambrosioides Leaves, possessing drug-likeness with satisfactory bioactivity scores, having good pharmacokinetic values. Metabolism and toxicities were further studied using FAME3, GLORY, and pred-hERG. Slight cardiotoxicity and cytotoxicity were predicted, respectively, for Caryophyllene oxide and Carvone oxide, 5-Isopropenyl-2-Methylenecyclohexanol. Good inhibitory activities of the three compounds against Bacterial, Fungal, and Cardiovascular receptor targets. Hence, this is a comprehensive in silico approach to evaluate D.ambrosioides Leaves main phytocompounds in the background of its potential in future drug development.

Assessing lead and cadmium tolerance of Chenopodium ambrosioides during micropropagation: an in-depth qualitative and quantitative analysis.

The tolerance of Chenopodium ambrosioides to some heavy metals under in vitro environment was thoroughly investigated. A micropropagation protocol was developed to facilitate the mass production of plants and to identify metals-tolerant species for potential use in the restoration of polluted areas. Nodal explants exhibited callus formation when treated with N6-benzyladenin (BA) (1.5 mg/l) and a combination of BA/α-naphthalene acetic acid (NAA) at concentrations of 1.5/1.0 mg/l on the Murashige and Skoog (MS) medium. The optimal shoot formation was achieved with the callus grown on a medium enriched with 1.5/1.0 mg/l BA/NAA, resulting in an impressive number (21.89) and length (11.79 cm) of shoots. The in vitro shoots were rooted using NAA (1.0 and 1.5 mg/l) and were acclimatized in pots with 71% survival rate. After standardizing micropropagation protocol, the in vitro shoots were subjected to various doses of lead nitrate (Pb(NO3)2 and cadmium chloride (CdCl2). Pb(NO3)2 and CdCl2 in the media let to a reduction in shoot multiplication, decreasing from 18.73 in the control group to 11.31 for Pb(NO3)2 and 13.89 for CdCl2 containing medium. However, Pb(NO3)2 and CdCl2 promoted shoot length from 5.61 in the control to 9.86 on Pb(NO3)2 and 12.51 on CdCl2 containing medium. In the case of Pb(NO3)2 treated shoots, the growth tolerance index (GTI) ranged from117.64% to 194.11%, whereas for CdCl2 treated shoots, the GTI ranged from 188.23% to 264.70%. Shoots treated with high level of Pb(NO3)2induced reddish-purple shoots, while a low level of Pb(NO3)2 induced shoots displayed both green and reddish-purple colors in the same explants. In CdCl2 treated culture, the toxic effects were narrow leaf lamina, elongated petiole and a dark reddish purple coloration. These findings highlight the remarkable potential of C. ambrosioides to maintain growth and organogenesis even in the presence Pb(NO3)2 and CdCl2 on the MS medium, indicating a high degree of metal tolerance.

Transcriptomic, epigenomic and physiological comparisons reveal key factors for different manganese tolerances in three Chenopodium ambrosioides L. populations.

Chenopodium ambrosioides is a manganese (Mn) hyperaccumulator that can be used for Mn-polluted soil phytoremediation. However, the mechanism of Mn tolerance of C. ambrosioides remains largely unknown. In this study, the key factors for Mn tolerance of C. ambrosioides was investigated from the aspects of DNA methylation pattern, gene expression regulation and physiological function. We found that the two genotypes of C. ambrosioides populations have differentiated tolerance to Mn stress (Mn-tolerant: CS and XC, Mn-sensitive: WH). Although there was no difference in Mn accumulation between two types under excess Mn, the biomass and photosynthetic systems were more severely inhibited in Mn-sensitive type, as well as suffering more serious oxidative damage. More differentially expressed genes (DEGs) were downregulated in the Mn-tolerant type, indicating that the Mn-tolerant type tends to inhibit gene expression to cope with Mn stress. DEGs related to metal transport, antioxidant system, phytohormone and transcription factors contribute to the tolerance of C. ambrosioides to Mn, and account for difference in Mn stress sensitivities between the Mn-sensitive and tolerant types. We also found that DNA methylation variation may help to cope with Mn stress. The global DNA methylation level in C. ambrosioides increased under Mn stress, especially in the Mn-sensitive type. Dozens of methylated loci were significantly associated with the Mn accumulation trait of C. ambrosioides, and some critical DEGs were regulated by DNA methylation. Our study comprehensively demonstrated the Mn tolerance mechanism of C. ambrosioides for the first time, and highlighted the roles of epigenetic modification in C. ambrosioides response to Mn stress. Our findings may contribute to elucidating the adaptation mechanism of hyperaccumulator to the heavy metal toxicity.

Kaempferitrin, a major compound from ethanol extract of Chenopodium ambrosioides, exerts antitumour and hepatoprotective effects in the mice model of human liver cancer xenografts.

OBJECTIVES：: This study aimed to investigate antitumour effect and possible toxicity of kaempferitrin, the major compound from ethanol extract of Chenopodium ambrosioides, in the mice model of human liver cancer xenografts. METHODS：: Forty mice bearing SMMC-7721 cells xenografts were divided into control group (not treated) and three groups orally administered with ethanol extract of C. ambrosioides, kaempferol (positive control) and kaempferitrin for 30 days. Antitumour effect was evaluated by measurement of tumour growth, histological examinations of tumours, flow cytometry detection of splenic CD19+ B lymphocytes and CD161+ Natural Killer cells, biochemical measurements of serum levels of tumour necrosis factor-α, interleukin-6, interferon-γ, malonaldehyde, 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azinobis-(3-ethylbenz thiazoline-6-sulphonate) radicals. Toxicity was evaluated by histological examinations of livers and measurements of serum levels of aspartate transaminase, alanine transaminase, total bilirubin, direct bilirubin, malonaldehyde and hepatic malonaldehyde level. KEY FINDINGS: Kaempferitrin significantly (P < 0.05) decreased tumour volume, mass and cell number. Antitumour effect was due to induction of tumour cells necrosis and apoptosis, stimulation of splenic B lymphocytes, decreases of radicals and malonaldehyde. Kaempferitrin did not change liver structure, and decreased serum levels of transaminases, bilirubin, malonaldehyde and hepatic malonaldehyde level. CONCLUSIONS: Kaempferitrin exerts antitumour and hepatoprotective effects.

Chemical Composition, Antioxidant and Antibacterial Activities and Acute Toxicity of Cedrus atlantica, Chenopodium ambrosioides and Eucalyptus camaldulensis Essential Oils.

The essential oils yield of Cedrus atlantica, Chenopodium ambrosioides and Eucalyptus camaldulensis was different. C. ambrosioides gave a relatively higher yield (2.1 ± 0.1%), while that of C. atlantica was low (1.0 ± 0.1%) and that of E. camaldulensis was lower (0.75 ± 0.1% of dry matter). The active ingredients of the essential oils and some of their biological effects were also determined. The characterization of their chemical compositions showed that the three essences have different chemical profiles: C. atlantica was richer in sesquiterpenes (ß-Himachalene (54.21%) and γ -Himachalene (15.54%)), C. ambrosioides was very rich in monoterpene peroxides and monoterpenes (α-Terpinene (53.4%), ascaridole (17.7%) and ρ-Cymene (12.1%)) and E. camaldulensis was very rich in monoterpene compounds and monoterpenols (p-cymene (35.11%), γ-Eudesmol (11.9%), L-linalool (11.51%) and piperitone (10.28%)). The in vitro measurement of antioxidant activity by the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) reduction assay showed a significant performance of the eucalyptus oil and average performance of the other two (C. atlantica and C. ambrosioides). The in vitro bio-test for their antimicrobial effects showed that the antibacterial activity differed depending on the essential oil and the concentration used, and that their bactericidal efficacy was similar or superior to that of synthetic antibiotics. The toxicity test on rats revealed that the LD50 of the three essential oils was 500 mg/kg body weight, which classifies them as category four cytotoxic natural products at high doses.

Genetic and epigenetic variation separately contribute to range expansion and local metalliferous habitat adaptation during invasions of Chenopodium ambrosioides into China.

BACKGROUND AND AIMS: Invasive plants often colonize wide-ranging geographical areas with various local microenvironments. The specific roles of epigenetic and genetic variation during such expansion are still unclear. Chenopodium ambrosioides is a well-known invasive alien species in China that can thrive in metalliferous habitats. This study aims to comprehensively understand the effects of genetic and epigenetic variation on the successful invasion of C. ambrosioides. METHODS: We sampled 367 individuals from 21 heavy metal-contaminated and uncontaminated sites with a wide geographical distribution in regions of China. We obtained environmental factors of these sampling sites, including 13 meteorological factors and the contents of four heavy metals in soils. Microsatellite markers were used to investigate the demographic history of C. ambrosioides populations in China. We also analysed the effect of epigenetic variation on metalliferous microhabitat adaptation using methylation-sensitive amplified polymorphism (MSAP) markers. A common garden experiment was conducted to compare heritable phenotypic variations among populations. KEY RESULTS: Two distinct genetic clusters that diverged thousands of years ago were identified, suggesting that the eastern and south-western C. ambrosioides populations in China may have originated from independent introduction events without recombination. Genetic variation was shown to be a dominant determinant of phenotypic differentiation relative to epigenetic variation, and further affected the geographical distribution pattern of invasive C. ambrosioides. The global DNA unmethylation level was reduced in metalliferous habitats. Dozens of methylated loci were significantly associated with the heavy metal accumulation trait of C. ambrosioides and may contribute to coping with metalliferous microenvironments. CONCLUSIONS: Our study of C. ambrosioides highlighted the dominant roles of genetic variation in large geographical range expansion and epigenetic variation in local metalliferous habitat adaptation.

Determinação do rendimento do óleo essencial de Chenopodium ambrosioides l. em função da variação sazonal / Determination of the income of the essential oil of chenopodium ambrosioides l. in function of the variations of temperature and pluviometric indexes in the period of 09/2006 until 08/2007 / Determinación del rendimiento de aceite esencial de Chenopodium ambrosioides l. en función de la variación estacional

Chenopodium ambrosioides conhecida como mastruço, é utilizado na medicina tradicional para tratamento de ascaridíase; controle de artrópodes e pragas domésticas; inibição do desenvolvimento de fungos do solo e de insetos, tratamento de lesões cutâneas e alívio da dor de barriga e gripe. O objetivo do presente trabalho constituiu na avaliação do teor extrativo do óleo essencial de Chenopodium ambrosioides L. cultivado no Horto Medicinal da Universidade Paranaense (Umuarama-PR), durante um ano, no período de setembro de 2006 a agosto de 2007. A metodologia foi realizada através da coleta do material vegetal para obtenção do óleo pelo processo de hidrodestilação dos frutos frescos de mastruço. O rendimento do óleo foi calculado a partir da massa dos frutos frescos pela massa do óleo essencial obtido. Também foram avaliados os parâmetros de temperatura e índices pluviométricos aferidos diariamente. Os resultados demonstram um maior rendimento do óleo no período em que as temperaturas e índices pluviométricos estavam mais elevados (1,096 % na primavera e 0,998 % no verão); já em meses com temperaturas mais amenas e/ou secos o rendimento de óleo essencial apresentou considerável redução (0,774 % no outono e 0,819 % no inverno). Isto indica que o aumento e diminuição da produção de óleo essencial pela planta é influenciada pelos fatores climáticos, mostrando a importância deste estudo com a finalidade de padronização da época de colheita para maior rendimento do óleo e consequente aplicação em produtos cosméticos e farmacêuticos.

Chenopodium ambrosioides known as mastruço, is used in traditional medicine to treatment ascariasis; control of arthropods and domestic pests; inhibition of the development of soil fungi and insects, treatment of skin lesions and relief of stomach ache and flu. The objective of the present work was to evaluation of the extractive content of the essential oil of Chenopodium ambrosioides L. cultivated in the Horto Medicinal da Universidade Paranaense (Umuarama-PR), during one year, from September 2006 to August 2007. The methodology was carried out through the collection of plant material to obtain the oil through the hydrodistillation process of fresh mastruço fruits. The oil yield was calculated from the mass of fresh fruits by the mass of essential oil obtained. Temperature parameters and rainfall measured daily were also evaluated. The results demonstrate a higher oil yield in the period when temperatures and rainfall were higher (1.096% in spring and 0.998% in summer); in months with milder and/or drier temperatures, the essential oil yield showed a considerable reduction (0.774% in autumn and 0.819% in winter). This indicates that the increase and decrease in the production of essential oil by the plant is influenced by climatic factors, showing the importance of this study with the purpose of standardizing the harvest time for greater oil yield and consequent application in cosmetic and pharmaceutical products.

Chenopodium ambrosioides conocido como mastruço, se utiliza en la medicina tradicional para tratar la ascaridiasis; control de artrópodos y plagas domésticas; inhibición del desarrollo de hongos e insectos del suelo, tratamiento de lesiones cutáneas y alivio del dolor de estómago y gripe. El objetivo del presente trabajo fue evaluar el contenido extractivo del aceite esencial de Chenopodium ambrosioides L. cultivado en el Horto Medicinal da Universidade Paranaense (Umuarama-PR), durante un año, de septiembre de 2006 a agosto de 2007. La metodología fue llevada a través de la recolección de material vegetal para la obtención del aceite mediante el proceso de hidrodestilación de frutos frescos de mastruço. El rendimiento de aceite se calculó a partir de la masa de frutos frescos por la masa de aceite esencial obtenido. También se evaluaron parámetros de temperatura y precipitación medidos diariamente. Los resultados demuestran un mayor rendimiento de aceite en el período de mayor temperatura y precipitaciones (1,096% en primavera y 0,998% en verano); en meses con temperaturas más suaves y/o más secas, el rendimiento de aceite esencial mostró una reducción considerable (0,774% en otoño y 0,819% en invierno). Esto indica que el aumento y disminución de la producción de aceite esencial por parte de la planta está influenciado por factores climáticos, mostrando la importancia de este estudio con el propósito de estandarizar el tiempo de cosecha para mayor rendimiento de aceite y consecuente aplicación en productos cosméticos y farmacéuticos.

Levantamento epidemiológico dos casos de tuberculose no Brasil e ações alternativas para auxiliar no tratamento / Epidemiological survey of tuberculosis cases in Brazil and alternative actions to aid in treatment / Estudio epidemiológico de los casos de tuberculosis en Brasil y acciones alternativas para ayudar al tratamiento

tuberculose é uma das doenças infectocontagiosas de maior importância no Brasil e no mundo. Afeta de forma importante populações em situação de vulnerabilidade social e econômica. O objetivo deste estudo foi realizar um levantamento do número de casos notificados de tuberculose no Brasil nos últimos 10 anos (2011 a 2021), avaliar os fatores que afetam a transmissão, bem como discutir o tratamento padrão e com fitoterápicos. O levantamento epidemiológico dos casos de tuberculose no Brasil de janeiro de 2011 a dezembro de 2021 foi realizado dentre os notificados pelo Sistema de Informação de Agravos de Notificação (SINAN). Os resultados indicaram um aumento linear de casos a partir de 2017 com 90.776 casos diagnosticados, em 2018 (94.720) e 2019 (96.655). Acredita-se que o aumento linear da tuberculose neste período pode estar relacionado principalmente com o aumento da pobreza, contudo o compartilhamento de utensílios durante o uso de narguilé podem representar fatores de risco para tuberculose. Seis plantas medicinais afetam diretamente as micobactérias (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata) e cinco plantas com atividade antibacteriana auxiliam no trato respiratório (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). Contudo, a tuberculose é uma doença reemergente sendo necessária a adoção de políticas públicas que intensifiquem e implementem medidas sócio-educativas para a implantação do uso de fitoterápicos como medida complementar.

Tuberculosis is one of the most important infectious diseases in Brazil and worldwide. It significantly affects populations in situations of social and economic vulnerability. This study aimed to survey the number of reported tuberculosis cases in Brazil in the last 10 years (2011 to 2021) to assess the factors that affect the transmission and discuss standard and herbal treatments. The epidemiological survey of tuberculosis cases in Brazil from January 2011 to December 2021 was carried out among those notified by the Notifiable Diseases Information System (SINAN). The results indicated a linear increase in cases from 2017, with 90,776 diagnosed cases, in 2018 (94,720) and 2019 (96,655). It is believed that the linear increase in tuberculosis in this period may be mainly related to the increase in poverty. However, the sharing of utensils during the use of hookah may represent risk factors for tuberculosis. Six medicinal plants directly affect mycobacteria (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata), and five plants with antibacterial activity help in the respiratory tract (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). However, tuberculosis is a re-emerging disease, and it is necessary to adopt public policies that intensify and implement socio-educational measures for using herbal medicines as a complementary measure.

La tuberculosis es una de las enfermedades infecciosas más importantes en Brasil y en el mundo. Afecta significativamente a las poblaciones en situación de vulnerabilidad social y económica. El objetivo de este estudio fue realizar una encuesta sobre el número de casos notificados de tuberculosis en Brasil en los últimos 10 años (2011 a 2021), para evaluar los factores que afectan a la transmisión, así como para discutir el tratamiento estándar y con fitoterapias. La encuesta epidemiológica de los casos de tuberculosis en Brasil desde enero de 2011 hasta diciembre de 2021 se realizó entre los notificados por el Sistema de Informação de Agravos de Notificação (SINAN). Los resultados indicaron un aumento lineal de casos desde 2017 con 90.776 casos diagnosticados, en 2018 (94.720) y 2019 (96.655). Se cree que el aumento lineal de la tuberculosis en este periodo puede estar relacionado principalmente con el aumento de la pobreza, aunque el hecho de compartir los utensilios durante el uso de la shisha puede representar factores de riesgo para la tuberculosis. Seis plantas medicinales afectan directamente a las micobacterias (Chenopodium ambrosioides, Tetradenia riparia, Physalis angulata, Origanum vulgare, Eucalyptus globulus, Mikania glomerata) y cinco plantas con actividad antibacteriana ayudan a las vías respiratorias (Nasturtium officinale, Allium sativum, Schinus terebinthifolius, Adiantum capillus-veneris, Allium cepa). Sin embargo, la tuberculosis es una enfermedad reemergente siendo necesaria la adopción de políticas públicas que intensifiquen e implementen medidas socioeducativas para la implementación del uso de fitoterápicos como medida complementaria.

Chemical composition and effect of Dysphania ambrosioides (L.) mosyakin & clemants essential oil on Biomphalaria tenagophila (D'Orbigny, 1835).

Dysphania ambrosioides (L.) Mosyakin & Clemants, known as santa maria herb, is a medicinal plant used on pest control and to treat parasitic diseases. Due to its potential for pest control, the aim of this work was to determine the chemical composition of D. ambrosioides essential oil and evaluate its molluscicidal effect on Biomphalaria tenagophila, which is an intermediate host for the Schistosoma mansoni trematode, that is responsible for causing human schistosomiasis. The essential oil extracted from its leaves showed 87% of (Z)-ascaridole and 100% of mortality against B. tenagophila at 10 ppm after 24 h of experiment. LC50 and LC90 values were, respectively, 2.40 (1.90-2.99) ppm and 8.75 (6.39-14.12) ppm. D. ambrosioides essential oil proved to be a promising natural molluscicide against schistosomiasis when compared with commercial products.

Phytochemical Analysis, Antispasmodic, Myorelaxant, and Antioxidant Effect of Dysphania ambrosioides (L.) Mosyakin and Clemants Flower Hydroethanolic Extracts and Its Chloroform and Ethyl Acetate Fractions.

Dysphania ambrosioides (L.) Mosyakin and Clemants is an annual or ephemeral perennial herb used traditionally in the Mediterranean region in folk medicine to treat various illnesses, including those related to the digestive system. This study aims to assess the antispasmodic, myorelaxant, and antioxidant effects of D. ambrosioides flower hydroethanolic extract and its chloroform and ethyl acetate fractions in a comparative study to evaluate the result of the extraction type on the potential activity of the extract. Both rat and rabbit jejunum were used to evaluate the antispasmodic and myorelaxant effect, while the antioxidant effect was evaluated using DPPH, a ferric reducing power assay, and a beta-carotene bleaching test. LC/MS-MS analysis was carried out to reveal the composition of the different types of extract. Following the results, the hydroethanolic extract showed a significant myorelaxant effect (IC50 = 0.39 ± 0.01 mg/mL). Moreover, it was shown that the hydroethanolic extract demonstrated the best antispasmodic activity (IC50 = 0.51 ± 0.05 mg/mL), followed by the ethyl acetate (IC50 = 4.05 ± 0.32 mg/mL) and chloroform (IC50 = 4.34 ± 0.45 mg/mL) fractions. The antioxidant tests showed that the hydroethanolic extract demonstrated high antioxidant activity, followed by the ethyl acetate and chloroform fractions. The LC/MS-MS analysis indicates that the plant extract was rich in flavonoids, to which the extract activity has been attributed. This study supports the traditional use of this plant to treat digestive problems, especially those with spasms.

Acropetal and basipetal cardenolide transport in Erysimum cheiranthoides (wormseed wallflower).

Plant specialized metabolites are often subject to within-plant transport and have tissue-specific distribution patterns. Among plants in the Brassicaceae, the genus Erysimum is unique in producing not only glucosinolates but also cardenolides. Ten cardenolides were detected with varying abundance in different tissues of Erysimum cheiranthoides L (Brassicaceae; wormseed wallflower). As is predicted by the optimal defense theory, cardenolides were most abundant in young leaves and reproductive tissues. The lowest concentrations were observed in senescing leaves and roots. Crosses between wildtype E. cheiranthoides and a mutant line with an altered cardenolide profile showed that the seed cardenolide phenotype is determined entirely by the maternal genotype. Prior to the development of the first true leaves, seedling cotyledons also had the maternal cardenolide profile. Hypocotyl grafting experiments showed that the root cardenolide profile is determined entirely by the aboveground plant genotype. In further grafting experiments, there was no evidence of cardenolide transport into the leaves, but a mixed cardenolide profile was observed in the stems and inflorescences of plants that had been grafted at vegetative and flowering growth stages, respectively. Together, these results indicate that E. cheiranthoides leaves are a site of cardenolide biosynthesis.

GC/MS analysis, anti-leishmanial and relaxant activity of essential oil of Chenopodium ambrosioides (L.) from Malakand region.

Chenopodium ambrosioides is abundantly available in Malakand region. As constituents and concentrations of essential oils vary based on its geographical location, we carried our current study to extract and evaluate its possible relaxant activity in rabbits' jejunum and anti-leishmanial activity against promastigotes of Leishmania tropica. The essential oil was obtained from aerial fresh parts through steam distillation followed by GC/MS analysis. Antispasmodic activity was performed on spontaneous and KCl induced contractions. Curves for calcium concentration response (CCRCs) were prepared with and without different concentrations of essential oils and verapamil - a standard calcium channel blocker as per our reported procedures. GC/MS analysis indicated that the essential oil contains 4-carene (56.59%) and o-cymene (41.46%), the two most abundant compounds previously reported from this species. The LD50 value for acute toxicity is 279.66±2.2mg/kg. The essential oil have significant antileishmanial activity with LC50 of Log10 (1.83±0.0026) ×10-6mg/ml, potent relaxant activity on rabbits' jejunal preparations with respective EC50 = 1.46±0.15mg/ml for spontaneous activity. For KCl (80mM) induced contractions, EC50=0.26±0.02mg/ml. In CCRCs, the oil produced a right shift as exhibited by verapamil. More, its relaxant activity, which is mediated through calcium channel blocking mechanism, proves a rationale for its traditional use in gut spasm.

Activity guided isolation and mechanistic approach towards analgesic potential of Chenopodium mediated through opioidergic pathway.

The current study is focused towards screening for its phytochemicals, phenolic and flavonoid contents of different species of Chenopodium. The plants were also screened for corroborating the traditional use of medicinal plants locally used for pain by determining the extract and their fractions for the in-vivo analgesic activity by using the modern scientific system. Among chloroform fractions, a high level of total phenolic contents was found in chloroform fraction of Chenopodium ambrosioides (ChAm-Chf) with 57.12±1.02 followed by Chenopodium botrys (ChBt-Chf) with 56.79±0.71. High content of flavonoids was found in chloroform fraction of Chenopodium botrys (ChBt-Chf) extract with 78.35±0.84 followed by Chenopodium ambrosioides (ChAm-Chf) with 75.20±0.81. The crude extract Chenopodium album, Chenopodium botrys and Chenopodium ambrosioides (ChAl-Crd, ChBt-Crd and ChAm-Crd) at 100 and 200 mg/kg, chloroform and ethylacetate fractions (ChAl-Chf, ChBt-Chf, ChAm-Chf, ChAl-Et, ChBt-Et and ChAm-Et) at 75 mg/kg caused significant inhibition (P<0.05, P<0.01, P<0.001, n=8) of the analgesic response induced by acetic acid, formalin and hotplate method. Mechanistically, the naloxone overturns completely the analgesic effects of beta-sitosterol (SN2) while partial reversal was observed by ursolic acid (SN1) indicating other possible mechanisms in association with opioid receptors.

Cytotoxicity and Apoptosis Induced by Chenopodium ambrosioides L. Essential Oil in Human Normal Liver Cell Line L02 via the Endogenous Mitochondrial Pathway Rather Than the Endoplasmic Reticulum Stress.

Chenopodium ambrosioides L. (C. ambrosioides) has been used as dietary condiments and as traditional medicine in South America. The oil of Chenopodium ambrosioides L. (C. ambrosioides) can be used as a natural antioxidant in food processing. It also has analgesic, sedating, and deworming effects, and can be used along with the whole plant for its medical effects: decongestion, as an insecticide, and to offer menstruation pain relief. This study was conducted to investigate the cytotoxicity and apoptosis effects of an essential oil from C. ambrosioides in vitro. The cytotoxicity evaluation of the essential oil from C. ambrosioides on human normal liver cell line L02 was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. AO/EB dual fluorescent staining assay and Annexin V-FITC were used for apoptosis analysis. The changes in mitochondrial membrane potential (MMP) were analyzed with 5,5,6,6'-tetrachloro-1,1,3,3,-tetraethyl-imidacarbocyanine iodide (JC-1) dye under a fluorescence microscope. The level of apoptosis related protein expression was quantified by Western blot. The L02 cells were treated with the essential oil from C. ambrosioides at 24, 48, and 72 h, and the IC50 values were 65.45, 58.03, and 35.47 µg/mL, respectively. The AO/EB staining showed that viable apoptotic cells, non-viable apoptotic cells, and non-viable non-apoptotic cells appeared among the L02 cells under the fluorescence microscope. Cell cycle arrest at the S phase and cell apoptosis increased through flow cytometry in the L02 cells treated with the essential oil. MMP decreased in a concentration-dependent manner, as seen through JC-1 staining under the fluorescence microscope. In the L02 cells as shown by Western blot and qPCR, the amount of the apoptosis-related proteins and the mRNA expression levels of cytochrome C, Bax, Caspase-9, and Caspase-3 increased, Bcl-2 decreased, and Caspase-12, which is expressed in the endoplasmic reticulum, showed no obvious changes in protein amount or mRNA expression level. The essential oil form C. ambrosioides had a cytotoxic effect on L02 cells. It could inhibit L02 cell proliferation, arrest the cell cycle at the S phase, and induce L02 cell apoptosis through the endogenous mitochondrial pathway.

The immunomodulatory activity of Chenopodium ambrosioides reduces the parasite burden and hepatic granulomatous inflammation in Schistosoma mansoni-infection.

ETHNOPHARMACOLOGICAL RELEVANCE: Folk medicine reports have described the use of Chenopodium ambrosioides as an anti-inflammatory, analgesic, and anthelmintic herb. These effects, including its activity against intestinal worms, are already scientifically observed. However, the immunological mechanisms of this species in the treatment of Schistosoma mansoni infection are unknown. AIM OF THE STUDY: To evaluate the immunological and anti-Schistosoma mansoni effects of a crude Chenopodium ambrosioides hydro-alcoholic extract (HCE). MATERIALS AND METHODS: For the in vitro analysis, cercariae and adult worms were exposed to different concentrations (0 to 10,000 µg/mL) of the HCE. For the in vivo evaluation, Swiss mice were infected with 50 cercariae of S. mansoni and separated into groups according to treatment as follows: a negative control (without treatment), a positive control (treated with Praziquantel®), HCE1 Group (treated with HCE during the cutaneous phase), HCE2 Group (treated with HCE during the lung phase), HCE3 Group (treated with HCE during the young worm phase), and HCE4 Group (treated with HCE during the adult worm phase). The animals treated with HCE received daily doses of 50 mg/kg, by gavage, for seven days, corresponding to the different developmental stages of S. mansoni. For comparison, a clean control group (uninfected and untreated) was also included. All animals were euthanized 60 days post-infection to allow the following assessments to be performed: a complete blood cells count, counts of eggs in the feces and liver, the quantification of cytokines and IgE levels, histopathological evaluations of the livers, and the analysis of inflammatory mediators. RESULTS: HCE treatment increased the mortality of cercariae and adult worms in vitro. The HCE treatment in vivo reduced the eggs in feces and liver. The number and area of liver granulomas, independent of the phase of treatment, were also reduced. The treatment with HCE reduced the percentage of circulating eosinophils, IgE, IFN-γ, TNF-α, and IL-4. In contrast, the treatment with the HCE, dependent on the phase, increased IL-10 levels and the number of peritoneal and bone marrow cells, mainly of T lymphocytes, B lymphocytes, and macrophages. This effect could be due to secondary compounds presents in this extract, such as kaempferol, quercetin and derivatives. CONCLUSIONS: This study demonstrates that Chenopodium ambrosioides has antiparasitic and immunomodulatory activity against the different phases of schistosomiasis, reducing the granulomatous inflammatory profile caused by the infection and, consequently, improving the disease prognosis.

Role of Ascaridole and p-Cymene in the Sleep-Promoting Effects of Dysphania ambrosioides Essential Oil via the GABAergic System in a ddY Mouse Inhalation Model.

The essential oil obtained from Dysphania ambrosioides leaves (DAEO) has antifungal, antioxidant, and antimicrobial properties. This study investigated DAEO's chemical composition and its sleep-promoting effects via administration by inhalation in ddY mice. Ascaridole (35.5%) and p-cymene (47.2%) were the major components. To obtain insight into DAEO's effects on the central nervous system (CNS), ascaridole and p-cymene were evaluated for sedative activity by using the caffeine-treated excitatory mouse model. DAEO administration significantly decreased locomotor activity at all doses except 0.000 04 mg per 400 µL of triethyl citrate. Both ascaridole and p-cymene were highly effective in decreasing locomotor activity of excited mice by more than 50%. In addition, ascaridole and p-cymene prolonged the pentobarbital-induced sleeping duration by 42% and 77%, respectively. These effects were antagonized by coadministration of gamma-aminobutyric acid (GABAA)-benzodiazepine receptor antagonist, flumazenil (3 mg/kg), indicating that the GABAergic system mediates the sedative effect. Finally, inhaled ascaridole and p-cymene had no negative effect on motor coordination, as observed during the Rota-rod test. Therefore, via activation of the GABAergic system, ascaridole and p-cymene mediate the sleep-promoting effect of DAEO. The results further extend the knowledge on their use as potential promising natural products for the management of sleep disorders and CNS-related ailments.

Effect of chitosan and Dysphania ambrosioides on the bone regeneration process: A randomized controlled trial in an animal model.

The focus of this triple-blind study was on evaluating the effect of chitosan combined with Dysphania ambrosioides (A) extract on the bone repair process in vivo. In total, 60 male Wistar rats (Rattus norvegicus albinus) weighing between 260 and 270 g were randomly selected for this study and distributed into four groups (n = 15). Group C (chitosan), Group CA5 (chitosan + 5% of D. ambrosioides), Group CA20 (chitosan + 20% of D. ambrosioides), and Group CO (Control-Blood clot). In each animal, bone defects measuring 2 mm in diameter were performed in both tibias for placement of the substances. After 7, 15, and 30 days, the animals were sedated and sacrificed using the cervical dislocation technique and the tissues were analyzed under optical microscope relative to the following events: inflammatory infiltrate, necrosis, osteoclasts, osteoblasts, fibroblasts, periosteal, and endosteal bone formation. The data were evaluated to verify distribution using the Kolmogorov-Smirnov test, and variance, using the Levene test; as distribution was not normal, data were subjected to the Kruskal-Wallis and Dunn nonparametric tests (p < .05). A significant inflammatory infiltrate was observed in Group CA5 (p = .008) in the time interval of 7 days, and in Group C at 15 (p = .009) and 30 (p = .017) days. Osteoblastic activity was more significant in Group CA20 (p = .027) compared with CA5 in the time interval of 7 days. Group CA20 demonstrated a significantly higher endosteal and periosteal bone formation value in the time interval of 7 (p = .013), 15 (p = .004), and 30 days (p = .008) compared with the other groups. The null hypothesis was refuted, bone regeneration was faster in spheres with an association of chitosan and 20% extract, and complete bone repair occurred clinically at 15 days and histologically at 30 days. The spheres proved to be a promising method for the biostimulation of alveolar bone repair and bone fractures.

Oxidative stability in raw, cooked, and frozen ground beef using Epazote (Chenopodium ambrosioides L.).

This study investigated the potential of aqueous epazote (AE) and ethanolic extract of epazote (ETHE) as inhibitors of lipid oxidation in raw and cooked ground beef stored at 4 °C for 9 days as well as frozen beef patties stored at -18 °C for 90 days. Organic acids were identified in AE and ETHE using ultra-high-performance liquid chromatography-quadrupole time-of-flight (UHPLC-qTOF). Chemical composition, TBARS, pH, colour, sensory acceptability and intensity of oxidised flavour were analysed in the three different meat products. ETHE showed higher values in organic acid content than AE. In raw ground beef, ETHE inhibited lipid oxidation and received the highest score in the three sensorial attributes evaluated at the end of the storage period, whereas in cooked ground beef it showed the highest intensity of oxidised flavour. Regarding CTL, AE reduced lipid oxidation in the cooked ground beef as well as the frozen patties, with improved colour. Therefore, epazote may be a promising natural antioxidant source for use in meat.

Chenopodium ambrosioides induces an endothelium-dependent relaxation of rat isolated aorta.

OBJECTIVE: This study aims to evaluate the vasodilatory effect of Chenopodium ambrosioides on the isolated rat aorta, and to explore its mechanism of action. METHODS: The vasorelaxant effect and the mode of action of various extracts from the leaves of C. ambrosioides were evaluated on thoracic aortic rings isolated from Wistar rats. In addition, ethyl acetate and methanol fractions were analyzed, using thin-layer chromatography and high-performance liquid chromatography techniques, for their polyphenolic content. RESULTS: The various active extracts of C. ambrosioides at four concentrations (10-3, 10-2, 10-1 and 1 mg/mL) relaxed the contraction elicited by phenylephrine, in a concentration-dependent manner. This effect seems to be endothelium-dependent, since the vasodilatory effect was entirely absent in denuded aortic rings. The vasorelaxant effect of the methanol fraction (MF) of C. ambrosioides at 1 mg/mL was also inhibited by atropine and tetraethylammonium. This effect remained unchanged by Nω-nitro-l-arginine methyl ester hydrochloride and glibenclamide. The preliminary phytochemical analysis showed that the leaves of C. ambrosioides are rich in phenolic and flavonoid derivatives. CONCLUSION: These results suggest that the MF of C. ambrosioides produces an endothelium-dependent relaxation of the isolated rat aorta, which is thought to be mediated mainly through stimulation of the muscarinic receptors, and probably involving the opening of Ca2+-activated potassium channels.

Antioxidant, intestinal immune status and anti-inflammatory potential of Chenopodium ambrosioides L. in fish: In vitro and in vivo studies.

Chenopodium ambrosioides L. has been used for centuries as traditional medicine in many clinical situations. The objectives of this study were first to assess the nutraceutical potential of C. ambrosioides L. extract through analyses of its chemical composition and antioxidant properties, followed by assessing toxicity and antioxidative activities on fish splenocytes. The second one was to perform an in vivo study using dietary C. ambrosioides L. extract (0.0, 0.5, 1.0 and 2.0%; w/w) for 15 and 30 days (2-week and 4-week treatments) to assess associated-intestine health status by short-chain fatty production, antioxidant enzyme activities and anti-inflammatory effects on Pacific red snapper (Lutjanus peru). Non-polar and polar fractions were detected by gas chromatography/mass spectrometry (GC-MS) in C. ambrosioides, of which the most abundant compounds were carvacrol, phytol, squalene, vitamin E and sucrose. The extract of C. ambrosioides L. enhanced a considerable antiradical and reducing power; fish splenocytes responded positively with higher (88%) cell viability than control. The production of nitric oxide and superoxide anion, as well as superoxide dismutase and catalase activities, were also enhanced in splenocytes treated with C. ambrosioides L. The in vivo study results showed that acetate was the major short-chain fatty acid found in fish receiving C. ambrosioides L. after week four. Pro-inflammatory cytokine gene expression in intestine was modulated in fish fed with C. ambrosioides L. at any time of the experimental trial. In addition, the histological findings suggested that its extract did not cause inflammatory damage in intestine. Overall, the results suggest that C. ambrosioides L. is safe for immune cells and promoting intestinal health status of fish through antioxidant and anti-inflammatory effects, making it an interesting additive in functional diets.