Ferulic acid as major antioxidant phenolic compound of the Tetragonisca angustula honey collected in Vera Cruz - Itaparica Island, Bahia, Brazil / Ácido ferúlico como principal composto fenólico antioxidante do mel de Tetragonisca angustula coletado em Vera Cruz - Ilha de Itaparica, Bahia, Brasil

Abstract The antioxidant activity of Tetragonisca angustula honey (TAH) and its ethanolic extract (TAEE) were investigated. The total levels of phenolic (TPC) and flavonoids (TFC) were also evaluated. The results for TPC were 19.91 ± 0.38 and 29.37 ± 1.82 mg GAE g-1 and for TFC 0.20 ± 0.02 and 0.14 ± 0.01 mg QE g-1 of TAH and TAEE, respectively. Antioxidant activities were 73.29 ± 0.49% and 93.36 ± 0.27% in the DPPH● assay and 71.73 ± 4.07% and 97.86 ± 0.35% in ABTS●+ for TAH and TAEE, respectively. The total reducing activity was determined by the method of reducing power (PR) and iron ion (Fe III) and the results varied in PR from 151.7 ± 25.7 and 230.7 ± 25.2 mg GAE L-1, for TAH and TAEE respectively and for (Fe III) in EC50 0.284 in TAEE and 0.687 in TAH. Chemical analysis by HPLC-DAD of the ethanolic extract (TAEE) revealed the presence of ferulic acid as majority phenolic component in the extract. The 1H NMR analysis confirmed this structure and showed the also presence of glucose, citric acid, succinic acid, proline and hydrocarbon derivatives. In addition, the botanical origin was also investigated and showed a multifloral characteristic, having found 19 pollen types with a botanical predominance of the Anacardiaceae family, with Tapirira pollen occurring as predominant (42.6%) and Schinus as secondary (25.7%). The results showed that T. angustula honey is an interesting source of antioxidant phenolic compounds due to its floral origin and can act as a protector of human health when consumed.

Resumo A atividade antioxidante do mel de Tetragonisca angustula (TAH) e seu extrato etanólico (TAEE) foram investigados. Os níveis totais de fenólicos (TPC) e flavonóides (TFC) também foram avaliados. Os resultados para TPC foram 19,91 ± 0,38 e 29,37 ± 1,82 mg GAE g-1 e para TFC 0,20 ± 0,02 e 0,14 ± 0,01 mg QE g-1 de TAH e TAEE, respectivamente. As atividades antioxidantes foram 73,29 ± 0,49% e 93,36 ± 0,27% no ensaio DPPH● e 71,73 ± 4,07% e 97,86 ± 0,35% no ABTS●+ para TAH e TAEE, respectivamente. A atividade redutora total foi determinada pelo método de poder redutor (PR) e íon ferrico (Fe III) e os resultados variaram em PR de 151,7 ± 25,7 e 230,7 ± 25,2 mg GAE L-1, para TAH e TAEE respectivamente e para (Fe III) em EC50 0,284 em TAEE e 0,687 em TAH. A análise química por HPLC-DAD do extrato etanólico (TAEE) revelou a presença de ácido ferúlico como componente majoritário no extrato. A análise de RMN 1H confirmou esta estrutura e mostrou a presença de glicose, ácido cítrico, ácido succínico, prolina e derivados de hidrocarbonetos no TAEE. Além disso, a origem botânica também foi investigada e apresentou característica multifloral, tendo encontrado 19 tipos polínicos com predomínio botânico da família Anacardiaceae, sendo o pólen Tapirira predominante (42,6%) e o Schinus secundário (25,7%). Os resultados mostraram que o mel de T. angustula é uma interessante fonte de compostos fenólicos antioxidantes devido à sua origem floral e pode atuar como protetor da saúde humana quando consumido.

Protective effects of sodium ferulate on flap transplantation via anti-inflammatory modulation and oxidative stress inhibition

Ischemia-reperfusion injury (IRI) has brought attention to flap failure in reconstructive surgery. To improve the prognosis of skin transplantation, we performed experimental IRI by surgical obstruction of blood flow and used sodium ferulate (SF) to prevent IRI in rats. After SF treatment, the morphological and histological changes of the skin flaps were observed by H&E and Masson's trichrome staining. We also detected the expression levels of COX-1, HO-1, and Ki67 by immunohistochemical and western blot analysis. Moreover, enzyme-linked immunosorbent assay was used to identify the content of tumor necrosis factor (TNF)-α, myeloperoxidase (MPO), malondialdehyde (MDA), and nitric oxide (NO) in peripheral blood and skin tissue. Compared with the model group, SF treatment significantly improved the recovered flap area (%) and promoted collagen synthesis. Cyclooxygenase-2 (COX-2) expression was significantly inhibited by heme oxygenase-1 (HO-1) induction after SF treatment. Furthermore, SF significantly inhibited the levels of TNF-α in peripheral blood, MPO and MDA in the skin tissue, and the increased synthesis of NO. Our results showed the protective effects of SF on IRI after flap transplantation and we believe that the protective effects of SF was closely related to the alleviation of the inflammatory response and the inhibition of the oxidative stress injury.

Ferulic acid improves motor function induced by spinal cord injury in rats via inhibiting neuroinflammation and apoptosis

ABSTRACT Purpose To investigate the effect of ferulic acid (FA) on spinal cord injury (SCI)-induced motor dysfunction and to explore the possible pharmacological mechanisms. Methods Adult male Wistar rats were used in our study. SCI was achieved by clipping the spinal cord T9 of the rat by a vascular clip for 2 minutes. The motor function of the rat was evaluated by Basso, Beattie, and Bresnahan scoring method (BBB) and inclined plane test. Hematoxylin and eosin (HE) staining, NISSL staining, and transmission electron microscopic examination were used to evaluate alterations at the histological level. Polymerase chain reaction (PCR), Western blots, and enzyme-linked immunosorbent assays (ELISA) were employed in biochemical analysis. Results The BBB score and inclined plane test score significantly decreased after SCI surgery, whereas chronic FA treatment (dose of 90 mg/kg, i.g.) for 28 days improved SCI-induced motor dysfunction. HE staining showed that SCI surgery induced internal spinal cord edema, but the structural changes of the spinal cord could be reversed by FA treatment. NISSL staining and transmission electron microscopic examination confirmed the improvement of the effect of FA on the injury site. In the biochemical analysis, it could be found that FA inhibitedSCI-induced mRNA and protein overexpression of pro-inflammatory cytokines (IL-1β, IL-6, TNF-α), as well as iNOS and COX-2 via the modulation of NF-κB level in the spinal cord of SCI rat. Moreover, the SCI-induced decrease of Bcl-2/Bax ratio was also reversed by FA treatment. However, the effect of FA on the expression of Beclin-1 was not statistically significant. Conclusions FA showed a therapeutic effect on SCI, which may be associated with the regulation of neuroinflammation and apoptosis.

Phytochemistry and Biological Activities of Werneria and Xenophyllum species / Fitoquímica y actividades biológicas de especies de Werneria y Xenophyllum

Plants of the genera Werneria (Asteraceae) and Xenophyllum (genus extracted from Werneria) are used in traditional medicine of Latin America for the treatment of mountain sickness, hypertension and gastrointestinal disorders. Only a small number of species of these genera have been studied, leading to the isolation of compounds belonging to the classes of benzofurans, chromenes, acetophenones, coumarates, diterpenes and pyrrolizidine alkaloids. Some of the plant extracts and/or compounds have shown antimicrobial, anti-HIV, hypotensive and photoprotective activities.

Las plantas de los géneros Werneria (Asteraceae) y Xenophyllum (género extraido de Werneria) son usadas en la medicina tradicional de América Latina para el tratamiento del mal de montaña, hipertensión y desórdenes gastrointestinales. Solo un pequeño número de especies de estos géneros ha sido investigado, lográndose aislar compuestos que pertenecen a las clases de benzofuranos, cromenos, acetofenonas, cumaratos, diterpenos y alcaloides pirrolizidínicos. Algunos de los extractos y/o compuestos de dichas plantas han mostrado actividades antimicrobianas, anti-HIV, hipotensoras y fotoprotectoras.

Metabolic engineering strategies for caffeic acid production in Escherichia coli

Caffeic acid (CA; 3,4-dihydroxycinnamic acid) is an aromatic compound obtained by the phenylpropanoid pathway. This natural product has antioxidant, antitumor, antiviral, and anti-inflammatory activities. It is also a precursor of CA phenethyl ester (CAPE), a compound with potential as an antidiabetic and liver-protective agent. CA can be found at low concentrations in plant tissues, and hence, its purification is difficult and expensive. Knowledge regarding the pathways, enzymes, and genes involved in CA biosynthesis has paved the way for enabling the design and construction of microbial strains with the capacity of synthesizing this metabolite. In this review, metabolic engineering strategies for the generation of Escherichia coli strains for the biotechnological production of CA are presented and discussed.

Type C feruloyl esterase from Aspergillus ochraceus: a butanol specific biocatalyst for the synthesis of hydroxycinnamates in a ternary solvent system

Background: Aspergillus ochraceus was isolated from coffee pulp and selected as an interesting hydroxycinnamoyl esterase strain producer, using an activity microplate high-throughput screening method. In this work, we purified and characterized a new type C A. ochraceus feruloyl esterase (AocFaeC), which synthesized specifically butyl hydroxycinnamates in a ternary solvent system. Results: AocFaeC was produced by solid state fermentation, reaching its maximal activity (1.1 U/g) after 48 h of culture. After purification, the monomeric protein (34 kDa) showed a specific activity of 57.9 U/mg towards methyl ferulate. AocFaeC biochemical characterization confirmed its identity as a type C feruloyl esterase and suggested the presence of a catalytic serine in the active site. Its maximum hydrolytic activity was achieved at 40°C and pH 6.5 and increased by 109 and 77% with Ca2+ and Mg2+, but decreased by 90 and 45% with Hg2+ and Cu2+, respectively. The initial butyl ferulate synthesis rate increased from 0.8 to 23.7 nmol/min after transesterification condition improvement, using an isooctane:butanol:water ternary solvent system, surprisingly the synthesis activity using other alcohols was negligible. At these conditions, the synthesis specific activities for butyl p-coumarate, sinapinate, ferulate, and caffeate were 87.3, 97.6, 168.2, and 234 U/µmol, respectively. Remarkably, AocFaeC showed 5 folds higher butyl caffeate synthesis rate compared to type B Aspergillus niger feruloyl esterase, a well-known enzyme for its elevated activity towards caffeic acid esters. Conclusions: Type C feruloyl esterase from A. ochraceus is a butanol specific biocatalyst for the synthesis of hydroxycinnamates in a ternary solvent system

Propolis from Southwest of Parana produced by selected bees: Influence of seasonality and food supplementation on antioxidant activity and phenolic profile

ABSTRACT Propolis produced by selected bees Apis mellifera were collected from March to June of 2013 and in March of 2015 and analyzed in order to evaluate the influence of climate, colony of origin, and food supplementation of colonies on the content of total phenolic and flavonoid by chromatographic analysis and antioxidant activity by radical scavenging of 2,2-diphenyl-1-picrylhydrazyl hydrate (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) and ferric reducing antioxidant power (FRAP) methods. The Principal Component Analysis (PCA) was carried out with propolis collected in 2013 and two clusters were formed. Propolis produced in the months of March and April showed a higher content of total phenolic compounds (TPC) and antioxidant capacity than those produced in May and June. The results of PCA obtained from samples collected in March of 2013 and 2015 showed two clusters, and propolis collected in 2015 were more bioactive and presented a higher content of TPC. The chromatographic analysis of extracts allowed the identification of phenolic acids p-coumaric, ferulic and caffeic with similar chemical profiles that could be closely related to the botanical origin of propolis. It can be concluded that the season and food supplementation of colonies influenced the chemical composition and the biological activity of samples analysed.

Dinámica de polifenoles en ephedra tweediana fisch & C. A. Mey. emend. J. H. Hunz. (Ephedraceae) / Polyphenol dinamics in ephedra tweediana fisch & C. A. Mey. emend. J. H. Hunz. (Ephedraceae)

Ephedra tweediana (Ephedraceae), conocida como “tramontana”, es empleada en la medicina popular como antiasmático. Se analizaron comparativamente los extractos acuosos de los tallos herbáceos, lignificados y partes subterráneas, provenientes de ejemplares femeninos y masculinos. En los tallos lignificados se determinó la presencia de proapigeninidina (cuya identidad fue corroborada por comparación con un testigo de apigeninidina sometido a estudios de FAB-MS, UV, HPLC, 1H-NMR y espectroscopía IR); mientras que en los órganos subterráneos se determinó la presencia de proapigeninidina y propelargonidina (la identidad de la pelargonidina fue establecida por comparación con un testigo de pelargonidina sometido a estudios de TLC/HPTLC y espectroscopía UV-Visible). Estos compuestos no se observaron en los tallos herbáceos. Los tallos herbáceos presentaron las mayores concentraciones de flavonoides y ácidos hidroxicinámicos totales. Los órganos subterráneos presentaron la mayor concentración de taninos y proantocianidinas. En los tallos herbáceos se detectó una reacción fuertemente positiva para flavonoides. No se observó reacción positiva para proantocianidinas. En el tallo aéreo lignificado se observó una reacción positiva para flavonoides y proantocianidinas a nivel de la peridermis. En los órganos subterráneos, los flavonoides y proantocianidinas se localizaron principalmente en los tejidos más externos. Este trabajo constituye el primer aporte a la dinámica de polifenoles de E. tweediana.

Ephedra tweediana (Ephedraceae), known as "tramontana" is used in folk medicine as antiasthmatic. Aqueous extracts obtained from young stems, woody stems and underground parts were analyzed and compared. In lignified stems was detected proapigeninidin (whose identity was confirmed by comparison with a control apigeninidin subjected to FAB-MS, UV, HPLC, 1H-NMR and IR spectroscopy), while underground organs were detected proapigeninidin and propelargonidin (by comparison with a control pelargonidin whose identity was established by studies of TLC/HPTLC, and UV-visible spectroscopy). These compounds were not observed in the herbaceous stems. The herbaceous stems had the highest concentrations of total flavonoids and hydroxycinnamic acids. Uderground organs had the highest concentration of tannins and proanthocyanidins. In herbaceous stems a strong positive reaction for flavonoids was detected. No positive reaction was observed for proanthocyanidins. In the periderm of woody aerial stem a positive reaction for flavonoids and proanthocyanidins were observed. In the underground organs, flavonoids and proanthocyanidins were located mainly in the more external tissues. This work is the first contribution to the dynamic of E. tweediana polyphenols.

Ferulic acid lowers body weight and visceral fat accumulation via modulation of enzymatic, hormonal and inflammatory changes in a mouse model of high-fat diet-induced obesity

Previous studies have reported on the glucose and lipid-lowering effects of ferulic acid (FA) but its anti-obesity potential has not yet been firmly established. This study investigated the possible anti-obesitogenic effects of FA in mice fed a high-fat diet (HFD) for 15 weeks. To assess the antiobesity potential of FA, 32 male Swiss mice, weighing 20–25 g (n=6–8 per group) were fed a normal diet (ND) or HFD, treated orally or not with either FA (10 mg/kg) or sibutramine (10 mg/kg) for 15 weeks and at the end of this period, the body weights of animals, visceral fat accumulation, plasma levels of glucose and insulin hormone, amylase and lipase activities, the satiety hormones ghrelin and leptin, and tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCH-1) were analyzed. Results revealed that FA could effectively suppress the HFD-associated increase in visceral fat accumulation, adipocyte size and body weight gain, similar to sibutramine, the positive control. FA also significantly (P<0.05) decreased the HFD-induced elevations in serum lipid profiles, amylase and lipase activities, and the levels of blood glucose and insulin hormone. The markedly elevated leptin and decreased ghrelin levels seen in HFD-fed control mice were significantly (P<0.05) reversed by FA treatment, almost reaching the values seen in ND-fed mice. Furthermore, FA demonstrated significant (P<0.05) inhibition of serum levels of inflammatory mediators TNF-α, and MCH-1. These results suggest that FA could be beneficial in lowering the risk of HFD-induced obesity via modulation of enzymatic, hormonal and inflammatory responses.

A stability-indicating HPLC-PDA method for the determination of ferulic acid in chitosan-coated poly(lactide-co-glycolide) nanoparticles

ABSTRACT The development and validation of a simple and efficient method for the quantification of ferulic acid in poly (D,L-lactide-co-glycolide) (PLGA) nanoparticles coated with chitosan (CS) by reverse phase high performance liquid chromatography coupled to photodiode array detection was described. For the chromatographic analysis, a reverse phase C-18 column was used, mobile phase consisting of acetonitrile and 0.5% acetic acid (37:63, v/v), isocratically eluted at a flow rate of 1 mL/min. Drug determination was performed at 320 nm. The method was validated in terms of the selectivity, linearity, precision, accuracy, robustness, limits of detection and quantification. The method was linear in the range of 10 to 100 µg/mL (r=0.999) and presented limit of detection and quantification of 102 ng/mL and 310 ng/mL, respectively. The method was precise (intra and inter-day) based on relative standard deviation values (less than 3.20%). The recovery was between 101.06 and 102.10%. Robustness was demonstrated considering change in mobile phase proportion. Specificity assay showed no interference from the components of nanoparticles or from the degradation products derived from acidic and oxidative conditions. The proposed method was suitable to be applied in determining the encapsulation efficiency of ferulic acid in PLGA-CS nanoparticles and can be employed as stability indicating one.

Comparative proteome analysis of Brettanomyces bruxellensisunder hydroxycinnamic acid growth

Background: Brettanomyces bruxellensis is an important spoilage yeast in the winemaking process. The capacity of this yeast to generate an undesired off-flavor constitutes a significant loss in the Chilean wine industry. Results: The proteomic profile of B. bruxellensis in the presence of p-coumaric acid was determined by 2D gel electrophoresis, gel image analysis and differential spot selection. A set of 41 proteins showed a differential accumulation of ±2 and a p-value <0.0001. The homology sequence analysis was performed using the databases available. Differential proteins belonged to the categories of 'energy production and conversion' and 'amino acid transport and metabolism'. Conclusions: The proteomic profile of B. bruxellensis cultivated in the presence of p-coumaric acid in synthetic wine, agrees with the hypothesis of metabolic flux regulation, allowing a better conditioning to an adverse environment. This study involved the translational level of B. bruxellensis in the production of ethylphenols and corroborated that this yeast presented an advantage in these stress conditions. Thus, this work will allow an understanding of the regulation and processes involved in the production of ethyl-derivate compounds by B. bruxellensis. Furthermore, it allows the development of newer and better techniques for spoilage yeast control.

Combined treatment with caffeic and ferulic acid from Baccharis uncinella C. DC. (Asteraceae) protects against metabolic syndrome in mice

Fractionation of the EtOH extract from aerial parts of Baccharis uncinella C. DC. (Asteraceae) led to isolation of caffeic and ferulic acids, which were identified from spectroscopic and spectrometric evidence. These compounds exhibit antioxidant and anti-inflammatory properties and have been shown to be effective in the prevention/treatment of metabolic syndrome. This study investigated whether the combined treatment of caffeic and ferulic acids exhibits a more significant beneficial effect in a mouse model with metabolic syndrome. The combination treatment with caffeic and ferulic acids was tested for 60 days in C57 mice kept on a high-fat (40%) diet. The data obtained indicated that treatment with caffeic and ferulic acids prevented gain in body weight induced by the high-fat diet and improved hyperglycemia, hypercholesterolemia and hypertriglyceridemia. The expression of a number of metabolically relevant genes was affected in the liver of these animals, showing that caffeic and ferulic acid treatment results in increased cholesterol uptake and reduced hepatic triglyceride synthesis in the liver, which is a likely explanation for the prevention of hepatic steatosis. In conclusion, the combined treatment of caffeic and ferulic acids displayed major positive effects towards prevention of multiple aspects of the metabolic syndrome and liver steatosis in an obese mouse model.

Sugars, organic acids, and phenolic compounds of ancient grape cultivars (Vitis vinifera L.) from Igdir province of Eastern Turkey

BACKGROUND: The Eurasian grapevine (Vitis vinifera L.) is the most widely cultivated and economically important horticultural crop in the world. As a one of the origin area, Anatolia played an important role in the diversification and spread of the cultivated form V. vinifera ssp. vinifera cultivars and also the wild form V. vinifera ssp. sylvestris ecotypes. Although several biodiversity studies have been conducted with local cultivars in different regions of Anatolia, no information has been reported so far on the biochemical (organic acids, sugars, phenolic acids, vitamin C) and antioxidant diversity of local historical table V. vinifera cultivars grown in Igdir province. In this work, we studied these traits in nine local table grape cultivars viz. 'Beyaz Kismis' (synonym name of Sultanina or Thompson seedless), 'Askeri', 'El Hakki', 'Kirmizi Kismis', 'Inek Emcegi', 'Hacabas', 'Kerim Gandi', 'Yazen Dayi', and 'Miskali' spread in the Igdir province of Eastern part of Turkey. RESULTS: Variability of all studied parameters is strongly influenced by cultivars (P < 0.01). Among the cultivars investigated, 'Miskali' showed the highest citric acid content (0.959 g/l) while 'Kirmizi Kismis' produced predominant contents in tartaric acid (12.71 g/l). The highest glucose (16.47 g/100 g) and fructose (15.55 g/100 g) contents were provided with 'Beyaz Kismis'. 'Kirmizi Kismis' cultivar had also the highest quercetin (0.55 mg/l), o-coumaric acid (1.90 mg/l), and caffeic acid (2.73 mg/l) content. The highest ferulic acid (0.94 mg/l), and syringic acid (2.00 mg/l) contents were observed with 'Beyaz Kismis' cultivar. The highest antioxidant capacity was obtained as 9.09 μmol TE g-1 from 'Inek Emcegi' in TEAC (Trolox equivalent Antioxidant Capacity) assay. 'Hacabas' cultivar had the highest vitamin C content of 35.74 mg/100 g. CONCLUSIONS: Present results illustrated that the historical table grape cultivars grown in Igdir province of Eastern part of Turkey contained diverse and valuable sugars, organic acids, phenolic acids, Vitamin C values and demonstrated important antioxidant capacity for human health benefits. Further preservation and use of this gene pool will be helpful to avoid genetic erosion and to promote continued agriculture in the region.

Poly R decolorization and APPL production by Streptomyces violaceoruber and Streptomyces spiroverticillatus

Two mesophilic streptomycetes (S. violaceoruber and S. spiroverticillatus) were selected to study their Poly R-478 decolorization ability and lignocellulose solubilizing activity. Both strains were able to degrade Poly R-478 dye and ferulic acid during growth on a minimal salts medium. The Poly R-478 decolorizing activities of both strains were induced by adding different carbon sources to the culture media. S. violaceoruber could decolorize 63% of Poly R-478 after 24 h. Both strains could solubilize straw and produce acid-precipitable polymeric lignin (APPL) with different efficiency. From the major extracellular enzymes recovery of both strains on rice and wheat straw, we can predicate that the biodegradation process was partial indicating a possible utilization in biological delignification.

Optimization of laccase production by two strains of Ganoderma lucidum using phenolic and metallic inducers / Optimización de la producción de lacasa por dos cepas de Ganoderma lucidum utilizando inductores fenólicos y metálicos

Ganoderma lucidum (Curtis) P. Karst es un hongo causante de pudrición blanca, capaz de degradar la lignina de la madera y otros sustratos en los que crece. En este trabajo se evaluó la capacidad de dos cepas de esta especie de producir la enzima ligninolítica lacasa. Asimismo, se ensayó la inducción de esta enzima con diferentes compuestos fenólicos e iones metálicos, y se encontró que el ácido ferúlico y el cobre fueron los mejores inductores de la lacasa entre los agentes evaluados. También se encontró que los dos tipos de inductores (fenólicos y metálicos) producen distintos patrones electroforéticos de actividad lacasa. Las concentraciones óptimas de los inductores fueron establecidas mediante un diseño factorial. Se estimó la estabilidad térmica de la lacasa en un amplio rango de pH ácidos, y se comprobó que a pH más altos la enzima es más termoestable.

Ganoderma lucidum (Curtis) P. Karst is a white rot fungus that is able to degrade the lignin component in wood. The ability of two strains of this species to produce the ligninolytic enzyme laccase was assessed. After the evaluation of induction with heavy metals and phenolic compounds, it was found that among the tested substances, copper and ferulic acid are the best laccase inducers. It was also observed that the two types of inducers (phenolic and metallic) produce different electrophoretic patterns of laccase activity. Optimized concentrations of inducers were obtained through a factorial design and the thermal stability of optimized supernatants was studied at a wide range of acidic pH. We found that the enzyme is more thermostable at higher pH values.

p-Coumaric acid reduces high glucose-mediated impairment of endothelium-dependent relaxation in rat aorta / Ácido p-cumárico reduce el deterioro mediado por alta glucosa de la relajación dependiente de endotelio en aorta de rata

p-Coumaric acid (p-CA) is an ubiquitous plant metabolite with antioxidant, anti-inflammatory, and anticancer properties. The present study was designed to evaluate the preventive effects of p-CA on endothelium-dependent impairment produced by high glucose (HG) (D-Glucose 25 mM) in isolated rat thoracic aorta. Aortic rings obtained from male Sprague-Dawley rats were mounted in an organ bath and pretreated for 3 h with D-Glucose 5 mM, HG and HG plus p-CA (1, 10 and 100 uM). After this period of time endothelium-dependent relaxation was tested by cumulative addition of acetylcholine (ACh) in pre-contracted rings with phenylephrine (PE) (0.1 μM). p-CA elicited a moderate endothelium-dependent vasodilatory effect (Emax= 29.28 +/- 1.89 percent, N=6; pD2= 6.075 +/- 0.184, N=6). When aortic rings were pre-incubated for 3 h with HG, Emax for ACh decreased dramatically from 87.69 +/- 2.59 percent (N=6) to 40.54 +/- 1.78 percent (N=6). The negative effect of HG was partially reverted in rings co-incubated with p-CA in a concentration-dependent manner as shown for Emax values to each p-CA concentration: 1 uM (48.57 +/- 1.82 percent), 10 uM (60.81 +/- 1.80 percent) and 100 uM (64.51 +/- 1.80 percent). The action of p-CA was associated with a significant change in Emax. No statistical difference in pD2 was observed. Our results clearly show that p-CA protect ACh-induced endothelial-dependent relaxation affected by HG in isolated rat aortic rings.

El ácido p-cumárico (p-CA) es un metabolito ubicuo en plantas, con propiedades antioxidantes, anti-inflamatoria, y anticancerígenas. El presente estudio fue diseñado para evaluar los efectos preventivos de p-CA sobre la relajación dependiente de endotelio, deteriorada por niveles altos de glucosa (HG) (D-glucosa 25 mM) en aorta torácica aislada de rata. Los anillos aórticos obtenidos de ratas macho Sprague-Dawley se montaron en un baño de órganos y fueron pre-tratados durante 3 h con D-glucosa 5 mM, HG, y HG más de p-CA (1, 10 y 100 uM). Después de este período de tiempo se evaluó la relajación dependiente de endotelio mediante la adición acumulativa de acetilcolina (ACh) en anillos pre-contraídos con fenilefrina (PE) (0,1 uM). p-CA mostró un moderado efecto vasodilatador dependiente de endotelio (Emax = 29,28 +/- 1,89 por ciento, N = 6; pD2 = 6,075 +/- 0,184, N = 6). Cuando los anillos aórticos se pre-incubaron durante 3 h con HG, la Emax para ACh se redujo drásticamente desde 87,69 +/- 2,59 por ciento (N = 6) a 40,54 +/- 1,78 por ciento (N = 6). El efecto negativo de HG se revirtió parcialmente, de manera dependiente de concentración, en los anillos co-incubados con p-CA tal como lo muestra el valor de Emax para cada concentración de p-CA: 1 u M (48,57 +/- 1,82 por ciento), 10 uM (60,81 +/- 1,80 por ciento) y 100 uM (64,51 +/- 1,80 por ciento). La acción de p-CA se asoció con un cambio significativo en la Emax. No se observó diferencia estadísticamente significativa en el pD2. Nuestros resultados muestran claramente que p-CA protege la relajación dependiente de endotelio inducida por ACh, la cual es afectada por HG en anillos aislados de aorta de rata.

Effect of chlorocholine chlorid on phenolic acids accumulation and polyphenols formation of buckwheat plants

BACKGROUND: Effect of chlorocholine chloride (CCC) on phenolic acids composition and polyphenols accumulation in various anatomical parts (stems, leaves and inflorescences) of common buckwheat (Fagopyrum esculentum Moench) in the early stages of vegetation period were surveyed. RESULTS: Treatment of buckwheat seeds with 2% of CCC has been increased content of total phenolics in the stems, leaves and inflorescences. On analyzing the different parts of buckwheat plants, 9 different phenolic acids - vanilic acid, ferulic acid, trans-ferulic acid, chlorogenic acid, salycilic acid, cinamic acid, p-coumaric acid, p-anisic acid, methoxycinamic acid and catechins were identified. The levels of identified phenolic acids varied not only significantly among the plant organs but also between early stages of vegetation period. Same changes as in contents of chlorogenic acid, ferulic acid, trans-ferulic acid were found for content of salycilic acid. The content of these phenolic acids has been significant increased under effect of 2% CCC treatment at the phase I (formation of buds) in the stems and at the phase II (beginning of flowering) in the leaves and then inflorescences respectively. The content of catechins as potential buckwheat antioxidants has been increased at the early stages of vegetation period after treatment with 2% CCC. CONCLUSIONS: The obtained results suggest that influence of CCC on the phenolics composition can be a result of various mechanisms of CCC uptake, transforming and/or its translocation in the buckwheat seedlings.

Induction of secondary metabolite production by UV-C radiation in Vitis vinifera L. Öküzgözü callus cultures

BACKGROUND: The aim of the present work was to examine the role of UV-C irradiation on the production of secondary metabolites (total phenolic, total flavanols, total flavonols, catechin, ferulic acid and trans-resveratrol in phenolic compounds and α-, ß-, γ- δ-tocopherols) in callus cultures. Studies on the effects of UV-C treatment on callus culture are seldom and generally focused on UV-B. However UV-C radiation play an important role in accumule secondary metabolites. RESULTS: In this study, callus cultures from Öküzgözü grape cultivar were initiated from leaf petiole explants. Calli formed after 6 weeks on the medium supplemented with 0.5 mg L-1 benzylaminopurine (BA), 0.5 mg L-1 indole acetic acid (IAA) on B5 media. Callus tissues were exposed to UV-C irradiation at 10, 20 and 30 cm distances from the UV source for 5 and 10 minutes and samples were collected at hours 0, 24 and 48. CONCLUSIONS: The greatest total phenolic content (155.14 mg 100 g-1) was detected in calli exposed to UV-C for 5 min from 30 cm distance and sampled after 24 h. 24 h and 48 h incubation times, 30 cm and 5 min were the most appropriate combination of UV-C application in total flavanol content. Maximum total flavonol content (7.12 mg 100 g-1) was obtained on 0 h, 5 min and 20 cm combination. The highest (+)- catechin accumulation (8.89 mg g-1) was found in calli with 10 min UV-C application from 30 cm distance and sampled after 48 h. Ferulic acid content increased 6 fold in Öküzgözü callus cultures (31.37 µg g-1) compared to the control group. The greatest trans-resveratrol content (8.43 µg g-1) was detected in calli exposed to UV-C for 5 min from 30 cm distance and sampled after 24 h. The highest α-tocopherol concentration was found in calli exposed to UV-C for 10 min from 30 cm distance and sampled after 24 h. As a conclusion, it was showed that UV-C radiation had remarkable promoting effects on the accumulation of secondary metabolites in the calli of Öküzgözü grape cultivar.

Molecular cloning, expression pattern, and putative cis-acting elements of a 4-coumarate: CoA ligase gene in bamboo (Neosinocalamus affinis)

Background: 4-coumarate:CoA ligase (4CL) plays an important role at the divergence point from general phenylpropanoid metabolism to several branch pathways. Although 4CL sin higher plants have been extensively studied, little has known about the 4CL gene of bamboo. Results: In current study, a Na4CL gene putative encoding 4-coumarate:CoA ligase (4CL) and its 5’-flanking region were isolated from bamboo (Neosinocalamus affinis) by RACE-PCR and genomic DNA walker, respectively. Na4CL encodes a predicted protein of 557 amino acids, with conserved motifs of adenylate-forming enzymes. Phylogenetic analysis showed that Na4CL shared 62~85 percent identity with other known plant 4CLs, and cluster closely with some known 4CLs in monocots. Sequence analysis revealed conserved cis-acting elements (Box A and AC-II element) present in the Na4CL promoter. Additionally, a Na4CL RNAi construct was transformed into tobacco. Transgenic tobaccos displayed significant down-expression of endogenesis 4CL and reduced lignin contents. Conclusion: These results contribute to the knowledge of the presence of Na4CL gene and its possible role in phenylpropanoid metabolism.

Validação de metodologia analítica para a determinação de derivados do ácido o-hidroxicinâmico de Echinodorus grandiflorus / Validation of analytical methodology for the determination of derivatives of o-hydroxycinnamic acid from Echinodorus grandiflorus

Este artigo descreve o desenvolvimento e a validação de método espectrofotométrico UV-Visível para quantificação de derivados do ácido o-hidroxicinâmico em folhas de Echinodorus grandiflorus. O método demonstrou ser linear (r² = 0,9974), preciso (DPR < 15%) na análise de matriz complexa e exata (recuperação = 107,56%).

This paper describes the development and the validation of an UV-Vis spectrophotometric method for the quantification of derivatives of o-hydroxycinnamic acid in leaves of Echinodorus grandiflorus. The method showed to be linear (r² = 0.9974), precise (RSD < 15%) in the analysis of complex matrix and accurate (recovery = 107.56%).