In vitro cytotoxic activity of Cuphea ingrata Cham. & Schltdl. extracts related to the oenothein B content.

Cuphea ingrata is a traditional medicinal plant species of the Lythraceae family. This work reports on the cytotoxic activity of the methanolic extract from the aerial parts of C. ingrata and the n-butanol and ethyl acetate fractions against human skin and prostate cancer cells. The selectivity of action was tested in normal skin keratinocytes HaCaT and prostate epithelial cells PNT2. The ethyl acetate fraction showed the highest activity in all three human skin cancer cell lines: A375, HTB-140, WM793, with IC50 = 15.90; 3.40; 18.75 µg/mL, respectively. To obtain comparative information on the chemical composition, a quantitative analysis of oenothein B was performed using the UHPLC-PDA method. An analysis of its cytotoxic activity was also carried out.

Polyphenolic Composition and in Vitro Antihypertensive and Anti-Inflammatory Effects of Cuphea lindmaniana and Cuphea urbaniana.

The present study investigates the chemical composition, anti-inflammatory, and antihypertensive activities, in vitro, from extracts of Cuphea lindmaniana and Cuphea urbaniana leaves. The extraction was performed ultrasound-assisted, and UHPLC/MS analysis was in positive mode ionization. The anti-inflammatory activity of the extracts and miquelianin were assayed at concentrations 0.001-10 µg/mL by chemotaxis on rat polymorphonuclear neutrophils. The antihypertensive activity was performed by angiotensin-converting enzyme (ACE) inhibition. From the nineteen proposed compounds, six of them are described for the first time in this genus. The extracts displayed antichemotactic effect with a reduction of 100 % of the neutrophil migration, in vitro, in most concentrations. The ACE-inhibition presented results ranging from 19.58 to 22.82 %. In conclusion, C. lindmaniana and C. urbaniana extracts contain a rich diversity of flavonoids and display in vitro anti-inflammatory and antihypertensive potential. Thus, this study could serve as a scientific baseline for further investigation, on developmental novel products with therapeutic actions.

Antinociceptive and anti-inflammatory effects of Cuphea aequipetala Cav (Lythraceae).

Cuphea aequipetala Cav (Lythraceae) is an herb used in folk treatment for pain and inflammation. The aim of this study was to evaluate the antinociceptive and anti-inflammatory actions of an ethanol extract from the leaves and stem of Cuphea aequipetala (CAE). The antinociceptive actions of CAE (10-200 mg/kg p.o.) were assessed with the acetic acid-induced writhing, hot plate, and formalin tests. The possible mechanism of action of CAE was evaluated using inhibitors. The effects of CAE on motor coordination were assessed by the rotarod test. The in vitro anti-inflammatory actions of CAE were evaluated using LPS-stimulated primary murine macrophages, and the in vivo anti-inflammatory actions were assessed by the TPA-induced ear oedema and the carrageenan-induced paw oedema tests. The production of inflammatory mediators was estimated from both in vitro and in vivo assays. CAE showed antinociception (ED50 = 90 mg/kg) in the acetic acid test and in the second phase of the formalin test (ED50 = 158 mg/kg). Pretreatment with glibenclamide or L-NAME partially reversed the antinociception shown by the plant extract. CAE (50-200 mg/kg) did not affect motor coordination in mice. CAE increased the production of IL-10 in LPS-stimulated macrophages (EC50 = 10 pg/ml) and, in the carrageenan-induced paw oedema test (threefold increase). In conclusion, CAE induced antinociceptive effects without affecting motor coordination, probably due to the involvement of nitric oxide and ATP-sensitive K+ channels. CAE also exerts in vitro and in vivo anti-inflammatory effects by increasing the release of IL-10.

Inhibition of biofilm and quorum sensing-regulated virulence factors in Pseudomonas aeruginosa by Cuphea carthagenensis (Jacq.) J. F. Macbr. Leaf extract: An in vitro study.

ETHNOPHARMCOLOGICAL RELEVANCE: Microbial biofilm formation, a quorum sensing (QS) regulated process, is one of the major causes of nosocomial and chronic infections, foodborne diseases, and associated deaths. Various approaches have been used to eradicate the menace of biofilm. Ethnomedicinal plants as potent antibiofilm agents are gaining a lot of interest in an era where the drug resistance is increasing and the availability of potent antibiotics is no longer promised. In this context, the methanol extract of Cuphea carthagenensis (CCMD), an ethno-medicinal and culinary herb, was evaluated as an antibiofilm and anti-QS agent against Pseudomonas aeruginosa. AIM OF THE STUDY: The aim of the study is to evaluate the antibiofilm and anti-QS activity of an ethnomedicinal plant against a strong biofilm forming microorganism, P. aeruginosa. METHODS: Antibiofilm activity of CCMD was demonstrated at different concentrations by Tissue Culture Plate, Test Tube method and other microscopic techniques. The effect of CCMD on QS and QS-related virulence factors viz. Pyocyanin, exopolymeric substance matrix (EPS), total protease, elastase, pyoverdin and swimming motility in P. aeruginosa were also evaluated. Antioxidant activity (DPPH & FRAP), total phenolic and flavonoid content were also checked. In order to determine the composition of the extract HPLC analysis was also performed. RESULTS: In vitro study demonstrated a significant inhibition of biofilm formation (81.88 ± 2.57%) as well as production of QS-dependent virulence factors in P. aeruginosa. The extract also inhibited violacein production (83.31 ± 2.77%) in Chromobacterium violaceum which correlates with the reduction in QS-mediated virulence factors. The extract showed 64.79% ± 0.83% DPPH scavenging activity and reduction of ferricyanide complex (Fe3+) to the ferrous form (Fe2+) in DPPH and FRAP assay, respectively. Furthermore, the extract showed thermal stability and does not have any growth inhibitory effect on P. aeruginosa. The HPLC analysis demonstrated the presence of ellagic acid, ascorbic acid and hippuric acid in the extract. CONCLUSION: This work is the first to demonstrate that C. carthagenensis can attenuate biofilm formation and QS-mediated virulence factors of P. aeruginosa. Further investigation is required to use this ethnomedicinal plant (CCMD) as an important source of antibiofilm agents.

Ultrasound-assisted extraction optimization and validation of ultra-performance liquid chromatographic method for the quantification of miquelianin in Cuphea glutinosa leaves.

Cuphea glutinosa is a medicinal species abundant in South of Brazil, known because of its flavonoids, which have pharmacological properties as antioxidant, anti-hypertensive, diuretic, and antimicrobial. The present study aimed to optimize the extraction and validate an ultra-performance liquid chromatographic method coupled to a photodiode array detector (UPLC-PDA) method for the quantification of a chemical marker miquelianin in C. glutinosa leaves. The optimum conditions for the extraction of miquelianin from leaves of C. glutinosa were determined using a fractional factorial design (FFD) and a central composite design (CCD). An UPLC-PDA method was validated, following the ICH guidelines and RDC 166/2017 of ANVISA (Brazil). The extraction-optimization methodology was obtained with the following parameters: plant:solvent 1:60 (w/v), percentage solvent 38% ethanol, 60 min time, five extractions and particle size ≤ 180 µm. The validation parameters of the quantification method were satisfactory. The results revealed a method with excellent selectivity, linearity, precision (repeatability and intermediate precision were below 2.18 and 1.40%, respectively) and accuracy (mean recovery 90.6%). The average content of miquelianin was 1.03%. Briefly, the optimization of the extractive method in the leaves of C. glutinosa increased the concentration of miquelianin in the crude extract and the method was validated according to the current legislation.

Polyphenols composition from leaves of Cuphea spp. and inhibitor potential, in vitro, of angiotensin I-converting enzyme (ACE).

ETHNOPHARMACOLOGICAL RELEVANCE: Cuphea is the largest genus of the Lythraceae family. It is popularly known as "sete-sangrias" in Brazil used in folk medicine as a diuretic, antipyretic, anti-inflammatory, laxative and antihypertensive agent. The raw material of Cuphea has shown promising results in the production of fitotherapics, which are chemically characterized by quercetin core flavonoids. AIMS OF THE STUDY: Present work aims to investigate the chemical composition of Cuphea calophylla, Cuphea carthagenensis, Cuphea glutinosa and Cuphea racemosa by UHPLC-MS using ESI-Q-TOF, and also to investigate the inhibition of angiotensin-converting enzyme (ACE) in vitro. MATERIALS AND METHODS: Leaves extraction was conducted by an ultrasound-assisted system under the following conditions: 40% ethanol, particle size ≤180 µm, plant:solvent ratio 1:20 (w/v) for 30 min. The leaf extracts were analyzed by UHPLC-MS positive mode ionization. For the inhibition of ACE, the leaf extracts used were obtained from different Cuphea species collected from several regions of Rio Grande do Sul (Brazil). RESULTS: In total 26 polyphenolic compounds were proposed, which were mostly derived from quercetin, myricetin, and kaempferol. Of these compounds, ten are described in the genus for the first time. The ACE-inhibiting activities are presented in descending order: miquelianin (32.41%), C. glutinosa 1 (31.66%), C. glutinosa 5 (26.32%) and C. carthagenensis 1 (26.12%). CONCLUSION: The obtained results suggest that the ACE-inhibiting potential may be increased by the interactions among the different phytoconstituents present in the crude extract. These results corroborate with the popular usage of Cuphea genus as diuretic and antihypertensive agents in folk medicine.

Inhibitory Effect of Cuphea aequipetala Extracts on Murine B16F10 Melanoma In Vitro and In Vivo.

Cuphea aequipetala (C. aequipetala) has been used in Mexican traditional medicine since prehispanic times to treat tumors. In this paper, we evaluated the antiproliferative and apoptotic effect of the methanolic and aqueous extracts of C. aequipetala on several cancer cell lines including the B16F10 cell line of murine melanoma and carried a murine model assay. In vitro assay analyzed the effect in the cellular cycle and several indicators of apoptosis, such as the caspase-3 activity, DNA fragmentation, phosphatidylserine exposure (Annexin-V), and induction of cell membrane permeabilization (propidium iodide) in the B16F10 cells. In vivo, groups of C57BL/6 female mice were subcutaneously injected with 5x105 B16F10 cells and treated with 25 mg/mL of C. aequipetala extracts via oral. Aqueous and methanolic extracts showed a cytotoxic effect in MCF-7, HepG2, and B16F10 cell lines. The methanolic extract showed more antiproliferative effect with less concentration, and for this reason, the in vitro experiments were only continued with it. This extract was able to induce accumulation of cells on G1 phase of the cell cycle; moreover, it was able to induce DNA fragmentation and increase the activity of caspase-3 in B16F10 cells. On the other hand, in the murine model of melanoma, the aqueous extract showed a greater reduction of tumor size in comparison with the methanolic extract, showing an 80% reduction versus one of around 31%, both compared with the untreated control, indicating a better antitumor effect of C. aequipetala aqueous extract via oral administration. In conclusion, the in vitro data showed that both C. aequipetala extracts were able to induce cytotoxicity through the apoptosis pathway in B16F10 cells, and in vivo, the oral administration of aqueous extract reduces the melanoma tumoral mass, suggesting an important antitumoral effect and the perspective to search for effector molecules involved in it.

A phytochemical study of the Cuphea glutinosa from Southern Brazil: Na+,K+-ATPase activity inhibition and antioxidant properties.

This study investigated the antioxidant activity of Cuphea glutinosa (CG) and its effect on Na+, K+-ATPase from cardiac muscle. The ethanolic extract showed higher antioxidant capacity compared to aqueous and ethyl acetate fraction. Ethyl acetate fraction showed ß-sitosterol-3-O-ß-glucoside, kaempferol, quercetin, isoquercetin, gallic acid methyl ester, and gallic acid. The ethanolic extract also reduced the Na+,K+-ATPase activity. CG presented a promising antioxidant activity and inhibitory effect on the Na+, K+-ATPase activity, supporting biochemical evidences the popular use of this plant in the treatment of heart failure.

Caracterización de compuestos fenólicos del extracto metanólico de hojas y tallos de Cuphea ciliata Ruiz & Pav. (hierba del toro) / Characterization of phenolic compounds of the metallic extract of stems and leaves of Cuphea ciliata Ruiz & Pav. (hierba del toro)

Objetivos. Caracterizar los compuestos fenólicos presentes en el extracto metanólico de hojas y tallos de Cuphea ciliata Ruiz & Pav. "hierba del toro". Materiales y métodos. Se preparó un extracto metanólico de las hojas y tallos de Cuphea ciliata Ruiz & Pav. Se evaluó la solubilidad del extracto en solventes de alta polaridad, mediana polaridad y no polares. Se detectaron los componentes químicos del extracto metanólico mediante un tamizaje fitoquímico empleando gelatina, tricloruro férrico, reacción de Mayer, reacción de Shinoda, entre otros. Se realizó cromatografía en capa fina y reveladores cromatográficos. Finalmente, mediante el uso del espectrofotómetro UV/Vis se hizo la propuesta de las estructuras de los flavonoides presentes en el extracto metanólico. Resultados. El extracto metanólico presentó una mejor solubilidad en solventes de alta polaridad. El tamizaje fitoquímico dio resultados positivos para la presencia de taninos, compuestos fenólicos, flavonoides, glicósidos y quinonas. Se propuso la estructura química de siete flavonoides obtenidos del extracto estudiado. Conclusiones. Se caracterizó la posible estructura química de siete flavonoides presentes en el extracto metanólico de hojas y tallos de Cuphea ciliata Ruiz & Pav. "hierba del toro".

Objectives. Characterize the phenolic compounds present in the methanolic extract of leaves and stems of Cuphea ciliata Ruiz & Pav. "Bull grass." Materials and methods. A methanolic extract was prepared from the leaves and stems of Cuphea ciliata Ruiz & Pav. The solubility of the extract in solvents of high polarity, medium polarity and non-polar was evaluated. The chemical components of the methanolic extract were detected by phytochemical screening using gelatin, ferric trichloride, Mayer reaction, Shinoda reaction, among others. Thin layer chromatography, chromatographic developers were performed. Finally, through the use of the UV / Vis spectrophotometer, the proposal of the flavonoid structures present in the methanolic extract was made. Results. The methanolic extract showed a better solubility in solvents of high polarity. Phytochemical screening gave positive results for the presence of tannins, phenolic compounds, flavonoids, glycosides and quinones. The chemical structure of seven flavonoids obtained from the studied extract was proposed. Conclusions. The possible chemical structure of seven flavonoids present in the methanolic extract of leaves and stems of Cuphea ciliata Ruiz & Pav. "Bull grass" was characterized.

Redox regulation and NO/cGMP plus K+ channel activation contributes to cardiorenal protection induced by Cuphea carthagenensis (Jacq.) J.F. Macbr. in ovariectomized hypertensive rats.

BACKGROUND: One of the medicinal plants widely used by the population in the treatment of hypertension, atherosclerosis and circulatory disorders is Cuphea carthagenensis (Jacq.) J.F. Macbr. (Lythraceae), popularly known as 'sete sangrias', being found in Brazil, Hawaii and in South Pacific Islands. Despite the widespread use of this species by the population, its long-term antihypertensive and cardioprotective activities have not yet been scientifically evaluated. PURPOSE: To evaluate the possible cardioprotective effects of an ethanol-soluble fraction obtained from C. carthagenensis (ESCC) using ovariectomized hypertensive rats to simulate a broad part of the female population over 50 years of age affected by hypertension. In addition, the molecular mechanism that may be responsible for its cardiorenal protective effects was also explored. METHODS: Female Wistar rats were submitted to surgical procedures of bilateral ovariectomy and induction of renovascular hypertension (two-kidneys, one-clip model). The sham-operated group was used as negative control. ESCC was obtained and a detailed phytochemical investigation about its main secondary metabolites was performed. ESCC was orally administered at doses of 30, 100 and 300  mg/kg, daily, for 28 days, 5 weeks after surgery. Enalapril (15  mg/kg) was used as standard antihypertensive drug. Renal function was evaluated on days 1, 7, 14, 21 and 28. At the end of the experimental period, systolic, diastolic, mean arterial pressure and heart rate were recorded. The activity of the tissue enzymatic antioxidant system, thiobarbituric acid reactive substances, nitrotyrosine, nitrite, aldosterone and vasopressin levels, in addition to the activity of the angiotensin-converting enzyme were also evaluated. Additionally, vascular reactivity to acetylcholine, sodium nitroprusside, and phenylephrine, and the role of nitric oxide, prostaglandins, and K+ channels in the vasodilator response of ESCC on the mesenteric vascular bed were also investigated. RESULTS: ESCC-treatment induced an important cardiorenal protective response, preserving renal function and preventing elevation of blood pressure and heart rate in ovariectomized hypertensive rats. In addition, prolonged treatment with ESCC recovered mesenteric vascular reactivity at all doses used. This effect was associated with an important modulation of the antioxidant defense system with a possible increase in NO bioavailability. Additionally, NO/cGMP activation and K+ channel opening-dependent vasodilator effect was observed on the mesenteric vascular bed, indicating a potential mechanism for the cardiovascular effects of ESCC. CONCLUSION: A 28-days ESCC treatment reduces the progression of the cardiorenal disease in ovariectomized hypertensive rats. These effects seem to be involved with an attenuation of oxidative and nitrosative stress, affecting endothelial nitric oxide production and K+ channel opening in smooth muscle cells.

A coumarin with an unusual structure from Cuphea ignea, its cytotoxicity and antioxidant activities.

Phenolic metabolite profiling using two dimensional paper chromatographic analysis (2 DPC) was used for assaying the complex mixture of phenolics of an aqueous ethanol aerial part extract of Cuphea ignea (Lytheraceae). A coumarin with a rare structure, namely, 7-hydroxy 3-methoxy coumarin 5-O-ß-glucopyranoside was isolated from the investigated extract. The structure was elucidated by conventional methods and spectral analysis, including one and two dimensional NMR (1D and 2D NMR), as well as by interpretation of the spectra obtained by high resolution electrospray ionization mass technique (HRESIMS). The rare coumarin significantly inhibited reactive oxygen species production with an ED50 value of 6.31±1.64 µg/ml and 5.78±0.66 µg/ml as determined by the the free radical 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the oxygen radical absorption capacity (ORAC) assay respectively. The isolated coumarin presented a cytotoxic activity assessed by using the neutral red assay (NRU) against lung cancer cell line (H23) with IC50 of 40.38±2.75 µg/ml.

Two distinct domains contribute to the substrate acyl chain length selectivity of plant acyl-ACP thioesterase.

The substrate specificity of acyl-ACP thioesterase (TE) plays an essential role in controlling the fatty acid profile produced by type II fatty acid synthases. Here we identify two groups of residues that synergistically determine different substrate specificities of two acyl-ACP TEs from Cuphea viscosissima (CvFatB1 and CvFatB2). One group (V194, V217, N223, R226, R227, and I268 in CvFatB2) is critical in determining the structure and depth of a hydrophobic cavity in the N-terminal hotdog domain that binds the substrate's acyl moiety. The other group (255-RKLSKI-260 and 285-RKLPKL-289 in CvFatB2) defines positively charged surface patches that may facilitate binding of the ACP moiety. Mutagenesis of residues within these two groups results in distinct synthetic acyl-ACP TEs that efficiently hydrolyze substrates with even shorter chains (C4- to C8-ACPs). These insights into structural determinants of acyl-ACP TE substrate specificity are useful in modifying this enzyme for tailored fatty acid production in engineered organisms.

A propagation procedure for Cuphea aequipetala Cav. (Lythraceae) and antioxidant properties of wild and greenhouse-grown plants / Procedimiento para la propagación de Cuphea aequipetala Cav. (Lythraceae) y propiedades antioxidantes de plantas silvestres y crecidas en invernadero

Cuphea aequipetala Cav. (Lythraceae) is native to Mexico and is used in folk medicine to treat tumors. An efficient protocol for in vitro shoot proliferation and plant acclimatization of C. aequipetala was developed. Total phenolic compounds and flavonoids contents were determined in methanolic extracts of roots, stems, and leaves from wild and greenhouse-grown plants. Their antioxidant properties were compared using in vitro assays (scavenging of 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-Azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radicals, and reducing power in the phosphomolybdenum assay). This is the first report of a successful propagation procedure for C. aequipetala. These methods offer a viable approach for long-term in vitro conservation and proliferation of this species. C. aequipetala shoots maintained their proliferation capacity during long-term subculture (3 years). The propagated shoots can successfully acclimatize and grow to maturity, and they retain the ability to accumulate antioxidants.

Cuphea aequipetala Cav. (Lythraceae) es una planta nativa de México que se utiliza en la medicina tradicional para tratar tumores. En este trabajo se desarrolló un procedimiento para la proliferación de brotes y la aclimatización de plantas de C. aequipetala. Se determinó la concentración de compuestos fenólicos totales y de flavonoides en extractos metanólicos de raíces, tallos y hojas de plantas silvestres y crecidas en invernadero. Sus propiedades antioxidantes fueron comparadas utilizando ensayos in vitro (captura de radicales DPPH y ABTS y poder reductor por el ensayo de fosfomolibdeno). Este es el primer reporte exitoso sobre un procedimiento para la propagación de C. aequipetala. Este método ofrece una alternativa viable para la conservación a largo plazo y la proliferación de esta especie. Los brotes de C. aequipetala han mantenido su capacidad de multiplicación a largo plazo (tres años). Los brotes se convirtieron en plantas adultas aclimatadas, manteniendo su habilidad para acumular compuestos antioxidantes.

Etnobotánica, micrografía analítica de hojas y tallos y fitoquimica de Cuphea aequipetala Cav. (Lythraceae): una contribución a la farmacopea herbolaria de los Estados Unidos Mexicanos (FHEUM) / Ethnobotany, analytical micrograph of leaves and stems and phytochemistry of Cuphea aequipetala Cav. (Lythraceae): a contribution to the herbal pharmacopoeia of the United Mexican States (FHEUM)

Cuphea aquipetala Cav. (Hierba del cancer) is used in Mexican traditional medicine. In the present work, its most frequent medicinal applications were determined through interviews, revision of herbarium specimens, and bibliography. The quoted applications were: to treat pounds and wounds. For these purposes, the drug (leaf and stem) is prepared and administered as infusion, or as poultice. In order to provide data for inclusion of this species in the Herbal Pharmacopoeia of the United Mexican States (FHEUM) we describe its leaf architecture and diagnostic anatomical characters of drug,. The anatomical study revealed that the three types of trichomes observed can aid to differentiate this species from other Cuphea species. Anatomical variation was observed mainly in the midrib of the leaf. Leaf architecture is first described for this species, and provides the basis for future comparisons. In addition, the acetone-water extract of the aerial parts previously reported as cytotoxic activity against DU-145 (human prostate carcinoma) and HCT-15 (human colon carcinoma), showed the presence of polyphenolic compounds of the type of hydrolysable and condensed tannins, mannitol was isolated and identified.

Cuphea aquipetala Cav., Lythraceae (Hierba del cáncer) es utilizada en la medicina tradicional mexicana. En el presente trabajo se determinó mediante entrevistas, consultas de herbario y revisión bibliográfica sus aplicaciones médicas más frecuentes, a saber: para tratar golpes y heridas, para lo cual la droga (hoja y tallo) es preparada en infusión, o administrada en emplastos. Con la finalidad de aportar datos que ayuden a la inclusión de esta especie en la Farmacopea Herbolaria de los Estados Unidos Mexicanos (FHEUM), se describen la arquitectura foliar y los caracteres anatómicos de la droga. El estudio anatómico reveló que los tres tipos de tricomas reconocidos en la especie estudiada apoyan a diferenciarla de otras especies de Cuphea sp. Se observó variación anatómica principalmente en la nervadura central de la hoja. La arquitectura foliar se describe por primera vez para la especie y sienta las bases para futuras comparaciones. Por otra parte, el extracto acetona-agua de las partes aéreas que previamente ha sido reportado con actividad citotóxica contra células DU-145 (carcinoma de próstata humano) y HCT-15 (carcinoma de colon humano) mostró la presencia de compuestos polifenólicos del tipo de los taninos hidrolizables, taninos condensados, y se aisló manitol.

Chemical composition and vasodilatation induced by Cuphea carthagenensis preparations.

The aerial parts of Cuphea carthagenensis (Jacq.) J.F. Macbride (Lythraceae) are traditionally employed in Brazil to treat cardiovascular diseases. The aim of this study was to compare preparations of C. carthagenensis aerial parts (aqueous and ethanol extracts, together with derived fractions) with regard to their total phenolic contents and in vitro vasodilating activity. The main flavonoids found in the extracts were isolated and identified as quercetin derivatives. The extracts and fractions showed similar HPLC profiles with the presence of quercetin-5-O-ß-glucopyranoside, quercetin-3-O-α-arabinofuranoside and quercetin-3-sulfate in all of them, but marked differences in the contents of flavonoids, proanthocyanidins, tannis and total phenolics. Excepting the aqueous extract, all assayed preparations elicited vasodilatation on pre-contracted rat aortic rings in the range of pIC(50) 4.53±0.03 to 4.98±0.06. Polynomial regression analysis demonstrated the relationship between vasodilating activity and the contents of flavonoids (r(2)=0.5190), proanthocyanidins (r(2)=0.8016), tannins (r(2)=0.8041) and total phenolics (r(2)=0.6226), suggesting the participation of these compounds in the pharmacological effect and their potential use as chemical markers for the species.

Additional antiprotozoal constituents from Cuphea pinetorum, a plant used in Mayan traditional medicine to treat diarrhoea.

In addition to kaempferol and quercetin already found in the roots from Cuphea pinetorum, bioassay-guided fractionation of the crude extract of the aerial part of this species gave four flavonoid glycosides, quercetin-3-O-alpha-rhamnopyranoside, luteolin-7-O-beta-D-glucopyranoside, apigenin-7-O-alpha-L-rhamnopyranoside and apigenin-7-O-beta-D-glucopyranoside, as well as squalen and beta-sitosterol. In vitro antiamoebic and antigiardial activities of isolated compounds indicated that kaempferol is the principal antiprotozoal agent in C. pinetorum. Based on finding this antiprotozoal inhibitor, flavonoids were studied in order to elucidate structure-activity relationships. These data suggest that kaempferol may play an important role in antidiarrhoeal activity of C. pinetorum.

Cytotoxic activity of Cuphea aequipetala.

Cuphea aequipetala (Lytraceae) is a perennial plant that has been used in Mexican traditional medicine to treat different types of tumors since prehispanic times. In the present work the cytotoxic potential of different fractions from acetone-water extract from the whole plant was investigated using a sulforhodamine B assay. Fractions were subjected to a bioscreening assay using several cell lines: HEp-2 (human larynx carcinoma), HCT-15 (human colon cancer) and DU-145 (human prostate carcinoma). Colchicine was used as positive control. Data are presented as the dose that inhibited 50.0% control growth (ED50). The cytotoxic activity is selective since the ED50 is different for the three cell lines employed. The highest activity was seen against the DU-145 cell line. "E" and PB1 fractions had the highest cytotoxic activities with ED50 values of 0.418 and 2.40 microg/ml respectively, on the DU-145 cell line. The "E" fraction was a yellow powder; it was methanol soluble and contained at least four separate components when separated by thin-layer chromatography. PB1 was a solid with metallic appearance; it was water soluble and its two dimensional chromatography showed 9 spots. These fractions have cytotoxic actives because their ED50 is less than 20 microg/ml and they will be further characterized.

The combined use of whole Cuphea seeds containing medium chain fatty acids and an exogenous lipase in piglet nutrition.

In search for an alternative for nutritional antimicrobials in piglet feeding, the effects of adding whole Cuphea seeds, as a natural source of medium chain fatty acids (MCFA), with known antimicrobial effects, and an exogenous lipase to a weaner diet were studied. The foregut flora, the gut morphology, some digestive parameters and the zootechnical performance of weaned piglets were investigated. Thirty newly weaned piglets, initial weight 7.0 +/- 0.4 kg, were divided according to litter, sex and weight in two groups (control diet; Cuphea + lipase diet). The Cuphea seeds (lanceolata and ignea) (50 g kg(-1)) were substituted for soybean oil (15 g kg(-1)), Alphacell (25 g kg(-1)) and soy protein isolate (10 g kg(-1)) in the control diet. Also 500 mg kg(-1) microbial lipase was added to the Cuphea diet. The piglets were weighted individually on days 0, 3. 7, 14 and 16. Feed intake was recorded per pen during days 0 to 3, 3 to 7, 7 to 14 and 14 to 16. On day 7 five piglets of each experimental group were euthanized for counting the gastric and small intestinal gut flora and for gut morphology at two sites of the small intestine (proximal, distal). The results indicate a trend towards improved performances parameters by feeding Cuphea + lipase. The enzymic released MCFA (1.7 g kg(-1) fresh gastric contents) tended to decrease the number of Coliforms in the proximal small intestine, but increased the number in the stomach and distal small intestine. With Culphea, the number of Streptococci was significantly lower in small intestine, but not in the stomach, while the number of Lactobacilli was significantly lower in the distal small intestine and tended to be lower in the stomach and proximal small intestine. No differences between the diets were noted for the total anaerobic microbial load in the stomach or in the gut. Feeding Cuphea + lipase resulted in a significantly greater villus height (distal small intestine) and a lesser crypt depth (proximal and distal small intestine) and greater villus/crypt ratio depth (proximal and distal small intestine). The intra-epithelial lymphocyte (IEL) counts per 100 enterocytes were significantly decreased in the proximal small intestine and tended to decrease in the distal small intestine by feeding the Cuphea + lipase diet. Both phenomena are indicative for a more healthy and better functional state of the mucosa. Present results are in line with foregoing research, showing that manipulation of the gut ecosystem by the enzymic in situ released MCFA in the stomach and foregut can result in improved performances of the piglets, which makes the concept a potential alternative for in-feed nutritional antibiotics.

In vitro immunomodulatory effects of cuphiin D1 on human mononuclear cells.

Cuphiin D1 (CD1), a macrocyclic hydrolyzable tannin isolated from Cuphea hyssopifolia, has been shown to exert antitumor activity both in vitro and in vivo. Moreover, the antitumor effects of CD1 are not only related to its cytotoxicity to carcinoma cell lines, but also depend on host-mediated mechanisms. In the present study, CD1 was investigated for its effects on the proliferation and cytokine secretion of human peripheral blood mononuclear cells (PBMCs). At concentrations of from 6.25 to 50 micrograms/ml, it enhanced the 3H-thymidine incorporation of concanavalin A (Con A)-stimulated PBMCs in a dose-dependent manner. Excretion of IL-1 beta, IL-2 and TNF-alpha by CD1-stimulated PBMCs was markedly increased in a dose-dependent manner. The results show that CD1 could stimulate PBMCs release of IL-1 beta, IL-2 and TNF-alpha and then activate T cells. Therefore, CD1-activated T cells via IL-1 beta in vitro might account for the host-mediated CD1 mechanism of action.