Echinodorus macrophyllus: Acute toxicological evaluation of hydroxycinnamoyl derivatives from SF1 subfractions.

ETHNOPHARMACOLOGICAL RELEVANCE: Echinodorus macrophyllus (Kunth.) Micheli (Alismataceae), known as chapéu-de-couro in Brazil, is popularly used to treat inflammatory diseases. We have previously demonstrated a significant reduction in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20) and described that hydroxycinnamoyl derivatives present in SF1 (Fr20 subfraction) showed higher anti-inflammatory properties by mechanisms that include a reduction of TNF-α, IL-1ß, CKCL1/KC, LTB4, and PGE2 levels in exudate. AIM OF THE STUDY: This work describes the acute toxicological effect of SF1 subfraction on SW mice treated orally for five days in the air pouch model by evaluating the hematological and biochemical determinations on the blood samples; the relative organ weight and its histopathological analysis; the liver genotoxicity assessment and the activity of liver enzymes from xenobiotic metabolism. MATERIALS AND METHODS: Fr20 was earlier fractionated on the Sephadex LH-20 column, yielding mainly four subfractions, including SF1. The SF1 toxicity was evaluated in mice challenged with carrageenan on the air pouch inflammation model and orally treated for five days. The body weight was monitored daily, and the organs were weighed after the euthanasia. Hematological and biochemical determinations were carried out using specific commercial kits and following the protocols provided by the manufacturers. The organs were fixed, sectioned, processed for hematoxylin and eosin staining, and analyzed by light microscopy. Genotoxicity assessment was performed by the alkaline single-cell gel electrophoresis. Livers were processed for ethoxyresorufin-O-deethylase (EROD) and Glutathione S-transferase (GST) assays. RESULTS: SF1 exhibited low toxicity, as no significant discrepancy was observed in the relative weight of the body organs of mice. Moreover, the daily treatment with SF1 did not alter the number and percentage of red blood cells or hemoglobin concentration in the blood. The treatment with SF1 did not affect the creatinine concentration, but the 25 mg/kg dose reduced the plasma urea level and uric acid, suggesting its use in treating acute renal failure. The parameters analyzed did not present biochemical alterations indicative of liver disease. Regarding serum triglyceride and cholesterol levels, a significant decrease was detected in both parameters in mice treated with SF1. In addition, the histopathological analysis showed that inflammatory focus in the livers seemed more relevant in the control groups than in those treated. There were no significant changes in the renal or splenic tissues of animals treated with SF1. Treatment with SF1 also does not have a genotoxic effect on liver cells. CONCLUSION: Treatment with SF1 showed no toxicity in mice at doses equivalent to those recommended for humans, which provides evidence of the safety of the therapeutic use of this subfraction.

Nichima gen. nov. (Alismataceae) based on reproductive structures from the Oligocene-Miocene of Mexico.

PREMISE: Alismataceae, a sub-cosmopolitan family with ca. 17 genera and 113 species, is a large group of aquatic plants. Compression/impressions and bioinclusions of reproductive parts in amber support the documentation of the lineage in low-latitude North America. In Mexico, fossil aquatic plants have been infrequently documented. The new reproductive structures exhibit characteristics of Alismataceae, whose fossil record is mainly documented in the northern hemisphere through of fruits and seeds. METHODS: We described and compared 150 samples of reproductive structures preserved as impressions/compressions from the Oligocene Los Ahuehuetes locality in the state of Puebla, and two bioinclusions from the Miocene amber of Simojovel de Allende in the state of Chiapas, Mexico with extinct and extant taxa. Using a parsimony analysis based on 29 floral characters of 17 extant genera of the Alismataceae, we evaluated the relationship between the fossil material and potential living relatives. RESULTS: We discovered a new genus Nichima based on a perfect, actinomorphic flower with an expanded receptacle, three persistent sepals with multiple vasculatures, delicate and caducous petals, six stamens, and a gynoecium composed of three to more superior carpels, maturing into achenes. These characteristics resemble flowers of Alismataceae. Nichima represents an extinct member of the family, with two new species described here, Nichima magalloniae L. Hern., Cevallos-Ferriz et Hernández-Damián sp. nov. and Nichima gonzalez-medranoi L. Hern., Cevallos-Ferriz et Hernández-Damián, sp. nov. Their phylogenetic position suggests affinity with a clade that includes Baldiella, Echinodorus, and Alisma. CONCLUSIONS: Reproductive structures from the Cenozoic of Mexico support the identification of a new extinct genus, Nichima, evidencing the extensive history of Alismataceae in North America's low latitudes and suggesting a southern extension of the boreotropical flora.

Phylogeny of Alisma (Alismataceae) revisited: implications for polyploid evolution and species delimitation.

Alisma L. is a genus of aquatic and wetland plants belonging to family Alismataceae. At present, it is thought to contain ten species. Variation in ploidy level is known in the genus, with diploids, tetraploids and hexaploids recorded. Previous molecular phylogenetic studies of Alisma have generated a robust backbone that reveals important aspects of the evolutionary history of this cosmopolitan genus, yet questions remain unresolved about the formation of the polyploid taxa and the taxonomy of one particularly challenging, widely distributed species complex. Here we directly sequenced, or cloned and sequenced, nuclear DNA (nrITS and phyA) and chloroplast DNA (matK, ndhF, psbA-trnH and rbcL) of multiple samples of six putative species and two varieties, and conducted molecular phylogenetic analyses. Alisma canaliculatum and its two varieties known in East Asia and A. rariflorum endemic to Japan possess closely related but heterogeneous genomes, strongly indicating that the two species were generated from two diploid progenitors, and are possibly siblings of one another. This evolutionary event may have occurred in Japan. Alisma canaliculatum var. canaliculatum is segregated into two types, each of which are geographically slightly differentiated in Japan. We reconstructed a single phylogeny based on the multi-locus data using Homologizer and then applied species delimitation analysis (STACEY). This allowed us to discern A. orientale as apparently endemic to the Southeast Asian Massif and distinct from the widespread A. plantago-aquatica. The former species was most likely formed through parapatric speciation at the southern edge of the distribution of the latter.

Flowering Biology of Alisma plantago-aquatica (Alismataceae).

Field observations of flowering Alisma plantago-aquatica plants were carried out in Moscow region (Russia). The A. plantago-aquatica flower remains anthetic for a single light day, from 9:00 a.m. to 8:00 p.m. White petals showed a contrast bicolored pattern in UV light, and the pattern probably serves as a nectar clue for pollinators. Flowers were visited by insects in daytime from 11:00 a.m. to 3:00 p.m. Coleopterans (Coccinellidae), dipterans (Drosophilidae, Hybotidae, Muscidae, Sepsidae, and Syrphidae), and hymenopterans (Apidae) were observed as flower visitors. Hoverflies (Syrphidae) and bees (Apidae) were the most frequent visitors. Large amounts of A. plantago-aquatica pollen grains were found on their bodies, and a major role in pollination was consequently assumed for the insects. Based on the original findings and literature data on A. plantago-aquatica reproductive biology in Belgium, Slovakia, and the Czech Republic, hoverflies were identified as the most stable and efficient pollinators of A. plantago-aquatica in various parts of the species range. Bees (Apidae) were recognized as A. plantago-aquatica pollinators for the first time in this work. A flower isolation experiment confirmed that A. plantago-aquatica is a self-compatible plant, but requires insects for the most efficient cross-pollination.

Effects of inorganic arsenic species on the antioxidant enzyme system of the Amazon Sword Plant (Echinodorus amazonicus Rataj).

This study aims to examine the effects of Arsenite (As+3) and Arsenate (As+5) on the aquatic macrophyte Amazon Sword Plant (Echinodorus amazonicus Rataj). To this aim, different concentrations of As+3 and As+5 (0, 6, 18 and 54 µM) were analyzed. At the end of the trail, photosynthetic pigment contents, total protein amounts, the enzymatic antioxidants superoxide dismutase (SOD), peroxidase (POX) and catalase (CAT) activities and the amount of malondialdehyde (MDA) in the leaf samples of E. amazonicus were investigated. The antioxidant enzyme activities increased at low concentrations (32.13% for SOD, 185% for CAT and 201.5% for POX in the groups of 6 µM As+5), but decreased at high concentrations (64.98% for SOD, 21.64% for CAT and 21.29% for POX in the groups of 54 µM As+3). MDA increased in all the treatment groups. The highest MDA contents were observed as 96% for 54 µM As+3 and 71.50% for 54 µM As+5. Photosynthetic pigment contents and the amount of protein were decreased with higher concentrations. The most significant decreases in protein content were 65% for 54 µM As+3 and 34.9% for 54 µM As+5. As a result, the toxicity of As+3 was higher and the toxic effect increased at higher concentrations.

Extrato de Echinodorus grandiflorus apresenta efeito renoprotetor em ratos com nefrotoxicidade aguda induzida pela ciclofosfamida / Extract of Echinodorus grandiflorus presents renoprotetic effect on rats with acute nephrotoxicity induced by cyclophosphamid / El extracto de Echinodorus grandiflorus tiene un efecto renoprotector en ratas con nefrotoxicidad aguda inducida por ciclofosfamida

Pelas características anatômicas e fisiológicas dos rins, a lesão renal aguda tem sua origem nefrotóxica pela alta circulação local, o que favorece a alta concentração de substâncias tóxicas e seus metabólitos no tecido. A lesão renal aguda é uma complicação comum em internações hospitalares e principalmente em internações em unidades de terapia intensiva. A ciclofosfamida, um quimioterápico utilizado no tratamento de doenças autoimunes e neoplasias sólidas, pode causar nefrotoxicidade com disfunção glomerular e tubular. O uso de plantas medicinais, pelas suas potentes ações antioxidantes, tem sido usado para prevenção ou tratamento de lesões celulares induzidas pelo desequilíbrio entre enzimas antioxidantes e oxidantes. Por esse motivo, o objetivo do experimento foi avaliar o potencial efeito protetor da Echinodorus grandiflorus na prevenção da nefrotoxidade induzida pela ciclofosfamida. Para isso, foi realizado o experimento com a utilização de 35 ratos machos, Wistar, divididos em seis grupos experimentais, sendo administrado a ciclofosfamida na dose de 150mg/kg nos grupos G2 a G6 e diferentes doses da Echinodorus grandiflorus, com posterior análise de parâmetros sanguíneos e histológicos. A administração de ciclofosfamida na dose de 150mg/kg de massa corporal, em dose única, foi capaz de induzir a nefrotoxicidade aguda em todos os ratos. O extrato bruto de Echinodorus grandiflorus apresentou potencial efeito renoprotetor ao uso da ciclofosfamida, na dose de 300mg/kg de massa corporal, sendo possível observar redução dos efeitos nefrotóxicos do quimioterápico, pela redução dos danos tubulares e pela diminuição dos espaços capsulas, nitidamente encontradas alterados no grupo que recebeu apenas ciclofosfamida, denotando resultados promissores para utilização desta planta medicinal na prevenção da nefrotoxicidade induzida pelo fármaco. Contudo, novos estudos dos efeitos renoprotetor do chapéu de couro, poderão elucidar os mecanismos envolvidos na ação do extrato bruto do chapéu de couro. A utilização de extrato bruto de plantas medicinais torna-se um adjuvante aos tratamentos pelo baixo custo e pela facilidade de acesso das diferentes populações as plantas desde que devidamente orientados pelos profissionais habilitados.

Due to the anatomical and physiological characteristics of the kidneys, acute kidney injury has its nephrotoxic origin due to the high local circulation, which favors the high concentration of toxic substances and their metabolites in the tissue. Acute kidney injury is a common complication in hospital admissions and especially in intensive care unit admissions. Cyclophosphamide, a chemotherapy drug used in the treatment of autoimmune diseases and solid neoplasms, can cause nephrotoxicity with glomerular and tubular dysfunction. The use of medicinal plants, due to their potent antioxidant actions, has been used for the prevention or treatment of cellular injuries induced by the imbalance between antioxidant and oxidant enzymes. For this reason, the aim of the experiment was to evaluate the potential protective effect of Echinodorus grandiflorus in preventing cyclophosphamide-induced nephrotoxicity. For this, the experiment was carried out with the use of 35 male Wistar rats, divided into six experimental groups, being administered cyclophosphamide at a dose of 150mg/kg in groups G2 to G6 and different doses of Echinodorus grandiflorus, with subsequent analysis of parameters blood and histology. The administration of cyclophosphamide at a dose of 150mg/kg of body weight, in a single dose, was able to induce acute nephrotoxicity in all rats. The crude extract of Echinodorus grandiflorus showed a potential renoprotective effect with the use of cyclophosphamide, at a dose of 300mg/kg of body mass, and it was possible to observe a reduction in the nephrotoxic effects of the chemotherapy, due to the reduction of tubular damage and the reduction of capsule spaces, clearly found altered in the group that received only cyclophosphamide, showing promising results for the use of this medicinal plant in the prevention of drug-induced nephrotoxicity. However, further studies of the renoprotective effects of the leather hat may elucidate the mechanisms involved in the action of the crude extract of the leather hat. The use of raw extract of medicinal plants becomes an adjuvant to treatments due to the low cost and ease of access of different populations to plants, provided that they are properly guided by qualified professionals.

Debido a las características anatómicas y fisiológicas de los riñones, la lesión renal aguda tiene su origen nefrotóxico por la elevada circulación local, que favorece la alta concentración de sustancias tóxicas y sus metabolitos en el tejido. La lesión renal aguda es una complicación frecuente en los ingresos hospitalarios y principalmente en las unidades de cuidados intensivos. La ciclofosfamida, un quimioterápico utilizado en el tratamiento de enfermedades autoinmunes y neoplasias sólidas, puede causar nefrotoxicidad con disfunción glomerular y tubular. El uso de plantas medicinales, debido a sus potentes acciones antioxidantes, se ha utilizado para la prevención o el tratamiento de lesiones celulares inducidas por el desequilibrio entre enzimas antioxidantes y oxidantes. Por este motivo, el objetivo del experimento era evaluar el posible efecto protector del Echinodorus grandiflorus en la prevención de la nefrotoxicidad inducida por la ciclofosfamida. Para ello, se realizó el experimento utilizando 35 ratas Wistar macho, divididas en seis grupos experimentales, administrándoseles ciclofosfamida a una dosis de 150mg/kg en los grupos G2 a G6 y diferentes dosis de Echinodorus grandiflorus, con posterior análisis de sangre y parámetros histológicos. La administración de ciclofosfamida a una dosis de 150mg/kg de masa corporal, en dosis única, fue capaz de inducir nefrotoxicidad aguda en todas las ratas. El extracto crudo de Echinodorus grandiflorus presentó un potencial efecto renoprotector al uso de ciclofosfamida, a una dosis de 300mg/kg de masa corporal, siendo posible observar una reducción de los efectos nefrotóxicos de la quimioterapia, por la reducción del daño tubular y por la disminución de los espacios capsulares, encontrándose claramente alterados en el grupo que recibió solamente ciclofosfamida, denotando resultados promisorios para el uso de esta planta medicinal en la prevención de la nefrotoxicidad inducida por el fármaco. Sin embargo, nuevos estudios sobre los efectos renoprotectores del sombrero de cuero podrían dilucidar los mecanismos implicados en la acción del extracto crudo de sombrero de cuero. El uso de extractos crudos de plantas medicinales se convierte en un coadyuvante de los tratamientos por su bajo coste y la facilidad de acceso de las diferentes poblaciones a las plantas desde que son guiadas adecuadamente por profesionales cualificados.

Efeito protetivo do extrato de Echinodorus grandiflorus sobre o urotelio vesical de ratos com cistite induzida por ciclofosfamida / Extract of echinodorus grandiflorus presents renoprotetic effect on rats with acute nephrotoxicity induced by cyclophosphamid / Extracto de Echinodorus grandiflorus presenta efecto renoprotético en ratas con nefrotoxicidad aguda inducida por ciclofosfamida

cistite hemorrágica e a cistite intersticial expressam uma etiologia variável, desde idiopática à provocada por fármacos, dentre eles a ciclofosfamida. A cistite apresenta tratamento multifatorial, e o potencial efeito satisfatório do uso da medicina complementar, vem ganhando espaço na prática médica. Assim o objetivo do presente estudo foi avaliar o efeito protetivo do extrato bruto de Echinodorus grandiflorus sobre a bexiga de ratos induzidos a cistite por ciclofosfamida. Utilizou-se neste estudo, 35 ratos, machos, Wistar, com peso médio de 321g, que foram submetidos a indução de cistite com uso de ciclofosfamida por via intraperitoneal e tratados com diferentes doses de extrato de Echinodorus grandiflorus (30, 100, 300mg) e o grupo controle com o fármaco Mesna. Todos os animais foram mortos no décimo sétimo dia e suas bexigas urinarias foram ressecadas para avaliação macro e microscópica, além da análise de hemograma e leucograma. A análise do sangue mostrou leucopenia com diferença significativa em todos os animais que receberam a ciclofosfamida. Observou-se que a dose de 300mg/kg do extrato bruto da planta, apresentou efeito protetivo no urotélio vesical, porém, inferior ao uso de Mesna. Diante dos resultados apresentados neste estudo sugere-se que o extrato de Echinodorus grandiflorus apresenta efeito protetivo no urotélio vesical na dose de 300mg/kg, porém estudos futuros quanto a dose e também a uma possível associação terapêutica ao Mesna devam ser realizados. Por se tratar de uma patologia com prevalência importante e ser muitas vezes desagradável e limitante à vida, faz-se necessário o empenho em métodos terapêuticos alternativos aos atuais, afim de, diminuírem os custos e efeitos colaterais dos métodos já documentados.

Hemorrhagic cystitis and interstitial cystitis have a variable etiology, from idiopathic to drug-induced, including cyclophosphamide. Cystitis has a multifactorial treatment, and the potential satisfactory effect of the use of complementary medicine has been gaining ground in medical practice. Thus, the aim of the present study was to evaluate the protective effect of the crude extract of Echinodorus grandiflorus on the bladder of rats induced to cystitis by cyclophosphamide. In this study, 35 male Wistar rats, with an average weight of 321g, were submitted to cystitis induction with intraperitoneal use of cyclophosphamide and treated with different doses of Echinodorus grandiflorus extract (30, 100, 300mg) and the control group with the drug Mesna. All animals were killed on the seventeenth day and their urinary bladders were resected for macro and microscopic evaluation, in addition to the analysis of blood count and leukogram. Blood analysis showed leukopenia with a significant difference in all animals that received cyclophosphamide. It was observed that the dose of 300mg/kg of the crude extract of the plant had a protective effect on the vesical urothelium, however, it was inferior to the use of Mesna. In view of the results presented in this study, it is suggested that the Echinodorus grandiflorus extract has a protective effect on the vesical urothelium at a dose of 300mg/kg, but future studies regarding the dose and also a possible therapeutic association with Mesna should be carried out. Because it is a pathology with significant prevalence and is often unpleasant and life-limiting, it is necessary to commit to alternative therapeutic methods to the current ones, in order to reduce the costs and side effects of the methods already documented.

cistitis hemorrágica y la cistitis intersticial tienen una etiología variable, desde idiopática hasta inducida por fármacos, incluida la ciclofosfamida. La cistitis tiene un tratamiento multifactorial, y el potencial efecto satisfactorio del uso de la medicina complementaria ha ido ganando terreno en la práctica médica. Así, el objetivo del presente estudio fue evaluar el efecto protector del extracto crudo de Echinodorus grandiflorus sobre la vejiga de ratas inducidas a cistitis por ciclofosfamida. En este estudio, 35 ratas Wistar macho, con un peso promedio de 321g, fueron sometidas a inducción de cistitis con uso intraperitoneal de ciclofosfamida y tratadas con diferentes dosis de extracto de Echinodorus grandiflorus (30, 100, 300mg) y el grupo control con el fármaco Mesna. Todos los animales fueron sacrificados al decimoséptimo día y sus vejigas urinarias fueron resecadas para evaluación macro y microscópica, además del análisis de hemograma y leucograma. El análisis de sangre mostró leucopenia con una diferencia significativa en todos los animales que recibieron ciclofosfamida. Se observó que la dosis de 300 mg/kg del extracto crudo de la planta tuvo un efecto protector sobre el urotelio vesical, sin embargo, fue inferior al uso de Mesna. En vista de los resultados presentados en este estudio, se sugiere que el extracto de Echinodorus grandiflorus tiene un efecto protector sobre el urotelio vesical a una dosis de 300 mg/kg, pero se deben realizar estudios futuros sobre la dosis y también una posible asociación terapéutica con Mesna. llevado a cabo. Por tratarse de una patología con una prevalencia importante y muchas veces desagradable y

cis-Aconitic Acid, a Constituent of Echinodorus grandiflorus Leaves, Inhibits Antigen-Induced Arthritis and Gout in Mice.

cis-Aconitic acid is a constituent from the leaves of Echinodorus grandiflorus, a medicinal plant traditionally used in Brazil to treat inflammatory conditions, including arthritic diseases. The present study aimed to investigate the anti-arthritic effect of cis-aconitic acid in murine models of antigen-induced arthritis and monosodium urate-induced gout. The possible underlying mechanisms of action was evaluated in THP-1 macrophages. Oral treatment with cis-aconitic acid (10, 30, and 90 mg/kg) reduced leukocyte accumulation in the joint cavity and C-X-C motif chemokine ligand 1 and IL-1ß levels in periarticular tissue. cis-Aconitic acid treatment reduced joint inflammation in tissue sections of antigen-induced arthritis mice and these effects were associated with decreased mechanical hypernociception. Administration of cis-aconitic acid (30 mg/kg p. o.) also reduced leukocyte accumulation in the joint cavity after the injection of monosodium urate crystals. cis-Aconitic acid reduced in vitro the release of TNF-α and phosphorylation of IκBα in lipopolysaccharide-stimulated THP-1 macrophages, suggesting that inhibition of nuclear factor kappa B activation was an underlying mechanism of cis-aconitic acid-induced anti-inflammatory effects. In conclusion, cis-aconitic acid has significant anti-inflammatory effects in antigen-induced arthritis and monosodium urate-induced arthritis in mice, suggesting its potential for the treatment of inflammatory diseases of the joint in humans. Additionally, our findings suggest that this compound may contribute to the anti-inflammatory effect previously reported for E. grandiflorus extracts.

Complete genus-level plastid phylogenomics of Alismataceae with revisited historical biogeography.

Alismataceae, an ancient lineage of monocots, has attracted attention due to its complex evolutionary history, ornamental value, and ecological role. However, the phylogenetic relationships and evolutionary history of the family have not been conclusively resolved. Here, we constructed the first complete genus-level plastid phylogeny of Alismataceae by using 78 genes and updated the historical biogeography based on the phylogenomic tree. Our results divide the Alismataceae into three major clades with robust support values; one clade comprises the former Limnocharitaceae, and the second clade includes the mainly temperate genera Alisma, Baldellia, Damasonium and Luronium, and the monotypic African genus Burnatia as a sister of the temperate genera. The remaining genera are either tropical or have some temperate species in addition to tropical ones, and they constitute the third major clade. Molecular dating and biogeographic analyses suggest that Alismataceae arose in Neotropical, West Palearctic, and Afrotropical regions during the Cretaceous, followed by the split into three main clades due to a combination of vicariance and dispersal events. Unlike earlier studies, we inferred that the mainly temperate clade likely originated from Afrotropical and West Palearctic regions during the Eocene. The most recent common ancestor of the other two clades lived in the Neotropical area during the Late Cretaceous. Long-distance dispersal and vicariance together seem to contribute to the transoceanic distribution of this family.

Echinodorus macrophyllus: Hydroxycinnamoyl derivatives reduces neutrophil migration through modulation of cytokines, chemokines, and prostaglandin in the air-pouch model.

ETHNOPHARMACOLOGICAL RELEVANCE: In Brazil, Echinodorus macrophyllus (Alismataceae), popularly known as chapéu-de-couro, is used to treat inflammatory diseases. Previous studies have shown a significant decrease in the acute inflammation for the aqueous extract of E. macrophyllus (AEEm) and its ethanolic fraction (Fr20). AIM OF THE STUDY: This work fractionated Fr20, identified the fraction and substances responsible for the in vivo anti-inflammatory property, and demonstrated important immunomodulatory mechanisms of action. MATERIALS AND METHODS: Fr20 was fractionated using Sephadex LH-20, and the most active fraction was chromatographically analyzed (HPLC-DAD and UPLC-ESI-TOF-MS). Leukotriene B4, Prostaglandin E2, and cytokines were determined by the enzyme-linked immunosorbent assay and in vivo acute inflammation by the air pouch model. RESULTS: The subfractions SF1, SF3, and mainly the SF4 decreased NO levels (p < 0.05). SF3 and SF4 showed high DPPH scavenger activity. SF1 was more effective than SF4 in reducing vasodilation, redness, and leukocyte migration into the 4-h air pouch. SF1 inhibited 90.5% (100 mg/kg) and SF4 54.0% (50 mg/kg), mainly affecting the number of neutrophils. SF1 and SF4 reduced the protein level in the exudate. SF1 was also more effective in inhibiting neutrophil migration in a transwell assay (46.3%) and reduced (86.1%) the Leukotriene B4 level in the exudate. After five days of treatment, some SF1 anti-inflammatory mechanisms were evaluated in the air pouch's 24 h exudate and tissue. Despite the high level of inflammation of the control group in this condition, SF1 confirmed the decrease in the protein level and neutrophils migration into the pouch. It decreased the number of bone marrow cells, indicating a systemic effect of SF1. SF1 also decreased TNF-α (87%), IL-1ß (77%), CKCL1/KC (71.3%), and PGE2 (97.8%) and increased IL-10 (74.1%) levels in the air pouch exudate. Phytochemical analysis of SF1 indicates mainly hydroxycinnamoyl derivatives. CONCLUSION: Hydroxycinnamoyl derivatives present in SF1 are related to the crucial anti-inflammatory mechanisms of E. macrophyllus, decreasing the levels of TNF-α, IL-1ß, CKCL1/KC, LTB4, and PGE2 on the exudate. These results explain the reduction of vasodilatation, erythema, and neutrophil migration into the air pouch model, confirming this plant's anti-inflammatory potential.

Improved structural annotation of triterpene metabolites of traditional Chinese medicine in vivo based on quantitative structure-retention relationships combined with characteristic ions: Alismatis Rhizoma as an example.

As a fast, sensitive and selective method, liquid chromatography-tandem high-resolution mass spectrometry (LC-HRMS) has been used for studying the in vivo metabolism of traditional Chinese medicine (TCM). However, the rapid discovery and characterization of metabolites, especially isomers, remain challenging due to their complexity and low concentration in vivo. This study proposed a strategy to improve the structural annotation of prototypes and metabolites through characteristic ions and a quantitative structure-retention relationship (QSRR) model, and Alismatis Rhizoma (AR) triterpenes were used as an example. This strategy consists of four steps. First, based on an in-house database reported previously, prototypes and metabolites in biosamples were preliminarily identified. Second, the candidate structures of prototype compounds and metabolites were determined by characteristic ions, databases or potential metabolic pathways. Then, a QSRR model was established to predict the retention times of the proposed structure. Finally, the structures of unknown prototypes and metabolites were determined by matching experimental retention times with the predicted values. The QSRR model built by the genetic algorithm-multiple linear regression (GA-MLR) has excellent regression correlation (R2 = 0.9966). Based on this strategy, a total of 118 compounds were identified, including 47 prototypes and 71 metabolites, among which 61 unknown compounds were reasonably characterized. The typical compound identified by this strategy was successfully validated using a triterpene standard. This strategy can improve the annotation confidence of in vivo metabolites of TCM and facilitate further pharmacological research.

Remediation of algal cells, PO43-, and NO3- from eutrophic wastewater using Echinodorus cordifolius in zigzag-horizontal subsurface constructed wetlands.

The pollutant removal efficiency of traditionally constructed wetlands (CWs) is often limited due to low interaction time between wastewater and the CW matrix (plants, microbes, and substrates). A zigzag-horizontal subsurface flow constructed wetland with effluent recirculation (Z-HSSF + ER) was developed to improve removal efficiency. Echinodorus cordifolius plants were used in this study. The efficiency of the systems was evaluated using eutrophic wastewater. The results showed that the developed systems exhibited the high removal efficiency of algal cells, PO43-, and NO3- (97%, 70%, and 100%, respectively), within 5 days. Algal cells were removed by the interception mechanism of gravel and zigzag baffles. PO43- and NO3- in the eutrophic wastewater was mainly removed by E. cordifolius including rhizobacteria and other microorganisms. The long flow pathway created by the installation of zigzag baffles combined with effluent recirculation provides high dissolved oxygen (DO) in the systems and increases the interaction time between wastewater and the CW matrix, thus improving the pollutant removal efficiency of CWs.

Bacteria Associated with Echinodorus cordifolius and Lepironia articulata Enhance Nitrogen and Phosphorus Removal from Wastewater.

Phytoremediation and bioremediation are eco-friendly methods of wastewater treatment that are widely used throughout the world to reduce anthropogenic water contamination. This study was conducted to assess the effectiveness of symbiotic bacteria in phytoremediation using two aquatic plants, Echinodorus cordifolius and Lepironia articulata, that were tested in sterilized and unsterilized groups. The results showed that unsterilized plants removed more phosphate, ammonium, nitrate and nitrite than the sterilized plants. In untreated and unsterilized E. cordifolius groups, the dominant bacterium was Calothrix (46.90 and 49.69%, respectively), which was higher than in the sterilized E. cordifolius group (38.88%). In untreated and unsterilized groups of L. articulata, Clostridium was a dominant bacterium. The proportion of Clostridium was much lower in the sterilized L. articulata group (1.31%) than in the untreated (13.71%) and unsterilized (49.02%) groups. Our results suggested that root-associated bacteria in E. cordifolius and L. articulata were effective in the removal of phosphorus and nitrogen from domestic wastewater.

Microvascular Effects of Echinodorus grandiflorus on Cardiovascular Disorders.

Echinodorus grandiflorus is a semiaquatic plant native to Brazil and belongs to the Alismataceae family. Infusion preparations of the leaves of this plant are often used due to its diuretic, blood pressure lowering, and anti-inflammatory properties. Our aim was to investigate the effects of chronic treatment with the crude hydroalcoholic extract of E. grandiflorus on central and peripheral microvascular changes induced in a model of hypertension and diabetes. The hemodynamic and microvascular effects of E. grandiflorus extract (50, 100, or 200 mg/kg/day for 28 days) or the isolated major diterpene from E. grandiflorus (3 to 10 mg/kg i. v.) were evaluated in spontaneously hypertensive rats using tail plethysmography and intravital fluorescence videomicroscopy, respectively, and were compared to vehicle-treated normotensive Wistar-Kyoto rats. We also investigated the protective effects of chronic treatment with E. grandiflorus (100 mg/kg/day) in brain capillary density and leukocyte-endothelium interactions on the brain vessels of DM-spontaneously (DM: diabetes mellitus) hypertensive rats. Chronically treating spontaneously hypertensive rats with increasing doses of crude hydroalcoholic E. grandiflorus extract resulted in significant dose-dependent reductions in systolic blood pressure and an anti-inflammatory effect on the brain microcirculation of DM-spontaneously hypertensive rat animals. Using laser speckle contrast imaging, we observed that intravenous administration of the major isolated clerodane diterpene metabolite (1 - 10 mg/kg) increased microvascular blood flow by 25% in spontaneously hypertensive rat skeletal muscle. The results of this study show that E. grandiflorus extracts can be useful in the prevention and reduction of microcirculatory damage in arterial hypertension and other diseases that involve microvascular dysfunction.

Root adaptation in Echinodorus osiris Rataj plant under cadmium stress.

Cadmium tolerant plant, Echinodorus osiris Rataj, was selected to study its root adaptive mechanism under Cd stress. The change of root porosity, radial oxygen loss (ROL), and iron plaque formation was investigated. Results suggested that Cd treatment decreased 28.6-49.9% of ROL and reduced 13.5-23.3% of root porosity but increased 63.4-147.2% of iron plaque after 21 days, respectively. Under different Cd treatments, the uptake of Cd in root presented quick and mild models while it showed relatively consistent increase in shoot. Correlation analysis demonstrated that Cd concentrations in plant were related negatively with root porosity but had no significant correlation with ROL. There was significant positive correlation between root porosity and ROL; however, they both related negatively with root iron plaque. Moreover, the scanning electron microscopy indicates a barrier to the movement of Cd in endodermis layers.

The complete chloroplast genomes of two species in threatened monocot genus Caldesia in China.

Caldesia is a genus in the family Alismataceae mainly found in the tropical and temperate regions of the Northern hemisphere. In China, two species, Caldesia parnassifolia, and Caldesia grandis are recorded as critically endangered in sporadic regions. Available protection of the genetic resource of these threatened species has been impeded due to limited genomic information. Here, we sequence the whole chloroplast (cp) genome of the two Caldesia species using high throughput sequencing technology. The whole cp genomes of C. parnassifolia and C. grandis were 167,647 bp and 168,500 bp, respectively with a typical quadripartite structure. There were 115 unique genes with 81 protein-coding genes, 31 tRNA genes, and four rRNA genes. Both species showed a GC content of 37.1%. A duplication of two tRNA genes and a ~ 6 kb inversion region in the LSC was noted in both species. Mononucleotide simple sequence repeats (SSRs) A/T were most abundant for both Caldesia species. High nucleotide variability was recorded in ycf1 gene and trnK-UUU/rps16 intergenic spacer region. All RNA editing conversions were C-U in 23 and 24 protein-coding genes for C. parnassifolia and C. grandis, respectively. Phylogenetic analysis placed both Caldesia species as sister to Sagittaria lichuanensis. This study will be useful for further evolutionary, systematic researches and conservation of the genus Caldesia.

Antiatherosclerotic Properties of Echinodorus grandiflorus (Cham. & Schltdl.) Micheli: From Antioxidant and Lipid-Lowering Effects to an Anti-Inflammatory Role.

Echinodorus grandiflorus is an important medicinal plant species that is native to South America. Despite extensive popular usage as a hypolipidemic drug, its effects as an atheroprotective agent remain unknown. The aim of this study was to evaluate the effects of an ethanol-soluble fraction that was obtained from E. grandiflorus (ESEG) leaves against the development of atherosclerosis in rabbits. Male rabbits received a diet that was supplemented with 1% cholesterol (cholesterol-rich diet [CRD]) for 60 days. After 30 days of the CRD, the animals were divided into five groups (n = 6) and treated with ESEG (10, 30, and 100 mg/kg), simvastatin (2.5 mg/kg), or vehicle once daily for 30 days. The negative control group was fed a cholesterol-free diet and treated orally with vehicle. At the end of 60 days, serum lipids, oxidized low-density lipoprotein, thiobarbituric acid reactive substances, nitrotyrosine, and serum interleukin 1 beta (IL-1ß), IL-6, soluble intercellular adhesion molecule-1 (sICAM-1), and soluble vascular cell adhesion molecule-1 (sVCAM-1) levels were determined. Samples from the aortic arch and thoracic segment were also collected to investigate the tissue antioxidant defense system and perform histopathological analysis. Oral ESEG administration significantly reduced serum lipid levels in CRD-fed rabbits. This treatment also modulated the arterial antioxidant defense system by reducing lipid and protein oxidation. Similarly, serum IL-1ß, IL-6, sICAM-1, and sVCAM-1 levels significantly decreased, accompanied by a reduction of atherosclerotic lesions in all arterial branches. These findings suggest that ESEG may be a new herbal medicine that can be directly applied for the treatment and prevention of atherosclerotic disease.

Increasing of electricity production from Echinodosus cordifolius-microbial fuel cell by inoculating Bacillus thuringiensis.

The wetland-microbial fuel cell (MFC) is a novel electricity generating technology. However, these systems can generate only limited electric energy. Since nitrification is a key mechanism driving electrical power in wetland-MFC systems, an effective nitrifying bacteria, Bacillus thuringiensis, was used to inoculate a wetland-MFC to enhance the maximum power density of the system. B. thuringiensis effectively enhanced the maximum power density, producing about 20-35 mW m-2 of maximum power density. Interestingly, over the first 120 days of operation, the wetland-MFC system with only B. thuringiensis generated more power than a system containing an Echinodosus cordifolius plant in addition to B. thuringiensis, because E. cordifolius can took up nitrate (NO3-) and phosphate (PO43-) in system's solution. Nitrate and PO43- act as important anions driving electric current in the system. After 120 days of operation though, the combined E. cordifolius and B. thuringiensis system maintained 20-35 mW m-2 maximum power density and the maximum power density of the system only inoculated with B. thuringiensis decreased continuously. Gene (16S rRNA) copy numbers for B. thuringiensis showed that when E. cordifolius was presented, the bacterium was able to continue growing after 120 days of operation. B. thuringiensis did not grow as well after 120 days in the system that did not contain a plant. This study presents a strategy for enhancing electric power output from a wetland-MFC by inoculating the system with B. thuringiensis and maintaining the bacterium's population with the support of an E. cordifolius plant. The result clearly show that B. thuringiensis can enhance electric power generation in the presence of the plant and the system can self-sustain for longer than 180 days of operation while producing 20-35 mW m-2 maximum power density.

Identification of the lipid-lowering component of triterpenes from Alismatis rhizoma based on the MRM-based characteristic chemical profiles and support vector machine model.

It has been demonstrated that triterpenes in Alismatis rhizoma (Zexie in Chinese, ZX) contributed to the lipid-lowering effect on high-fat diet-induced hyperlipidemia. Alisol B 23-acetate, one of the abundant triterpenes in ZX, was used as the marker of quality control for ZX in Chinese Pharmacopoeia, while it could not reflect the lipid-lowering effect because other triterpenes in ZX also had prominent medicinal efficacy. To identify the significantly bioactive triterpenes in ZX, a multiple reaction monitoring (MRM)-based characteristic chemical profile (CCP)-support vector machine (SVM) model was used to explore the relationship between triterpenes and lipid-lowering effect of ZX. Firstly, the content of 87 targeted triterpenes was quantified by the MRM-based CCP using UHPLC-QTRAP-MS/MS. Secondly, the lipid-lowering effect of 30 ZX samples was assessed by 3T3-L1 preadipocytes. Thirdly, 9 of the 87 triterpenes possessing high mean impact value were identified to have significant lipid-lowering effect via the particle swarm-optimized SVM model. The new SVM model constructed by the 9 triterpenes showed good prediction performance and the overall prediction accuracy reached 81.94%. Finally, the real activity of these triterpenes was partly confirmed and was consistent with the prediction of SVM. These results showed that the method for discovery of triterpenes with prominent lipid-lowering activity in ZX was reliable. The proposed method is expected to provide an efficient and rapid approach for screening of active component and drug discovery in traditional herbs. Graphical abstract.

Heart-Protective Effects of Echinodorus grandiflorus in Rabbits That Are Fed a High-cholesterol Diet.

Excess weight and dyslipidemia are among the most serious health problems in Western societies. These conditions enhance the risk of cardiac disease and have been linked with a higher prevalence of cardiac arrhythmias and sudden death. The present study investigated the cardioprotective effects of Echinodorus grandiflorus on ventricular remodeling in rabbits that were fed a 1% cholesterol-rich diet. We first obtained an ethanol-soluble fraction of E. grandiflorus and performed a detailed phytochemical study by liquid chromatography-DAD/ESI-MS. For 60 days, male rabbits were fed the cholesterol-rich diet or a diet without the addition of cholesterol. After 30 days, different groups of rabbits were treated with the ethanol-soluble fraction of E. grandiflorus (10, 30, and 100 mg/kg, p. o.), simvastatin (2.5 mg/kg), or vehicle once daily for 30 days. At the end of 60 days, the serum lipoprotein ratio, electrocardiographic profile, histopathological alterations, and the cardiac antioxidant defense system were investigated. Echocardiographic analysis showed morphological and functional alterations in cholesterol-rich diet-fed animals, indicating left ventricle hypertrophy. The total cholesterol/high-density lipoprotein ratio and low-density lipoprotein/high-density lipoprotein ratio were significantly higher in cholesterol-rich diet-fed rabbits. Myocardial flaccidity, fatty degeneration, and concentric left ventricular hypertrophy were observed. An increase in lipid peroxidation levels, a decrease in superoxide dismutase activity, and a decrease in reduced glutathione levels were observed in the myocardium of all cholesterol-rich diet-fed rabbits. Treatment with the ethanol-soluble fraction of E. grandiflorus, especially the highest dose, significantly reduced all of these alterations, thus demonstrating the cardioprotective effect of the ethanol-soluble fraction of E. grandiflorus on cardiac changes that are induced by a cholesterol-rich diet.