Ferulic acid-4-O-sulfate rather than ferulic acid relaxes arteries and lowers blood pressure in mice

Consumption of foods rich in ferulic acid (FA) such as wholegrain cereals, or FA precursors such as chlorogenic acids in coffee, is inversely correlated with risk of cardiovascular disease and type 2 diabetes. As a result of digestion and phase II metabolism in the gut and liver, FA is converted predominantly into ferulic acid-4-O-sulfate (FA-sul), an abundant plasma metabolite. Although FA-sul is the main metabolite, very little has been reported regarding its bioactivities. We have compared the ex vivo vasorelaxing effect of FA and FA-sul (10⁻⁷–3.10⁻⁵ M) on isolated mouse arteries mounted in tissue myographs. FA-sul, but not FA, elicited a concentration-dependent vasorelaxation of saphenous and femoral arteries and aortae. The FA-sul-mediated vasorelaxation was blunted by 1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one, a soluble guanylate cyclase (sGC) inhibitor. The role of sGC was confirmed in femoral arteries isolated from sGCα₁^(−/−) knockout mice. Furthermore, 4-aminopyridine, a specific inhibitor of voltage-dependent potassium channels, significantly decreased FA-sul-mediated effects. In anesthetized mice, intravenous injection of FA-sul decreased mean arterial pressure, whereas FA had no effect, confirming the results obtained ex vivo. FA-sul is probably one of the major metabolites accounting for the blood pressure-lowering effects associated with FA consumption.     

Groove binding between ferulic acid and calf thymus DNA: spectroscopic methodology combined with chemometrics and molecular docking studies

Abstract

Ferulic acid (FA), a dietary phenolic acid compound, is proved to possess numerous biological activities. Hence, this study was devoted to explore the interaction between FA and calf thymus DNA (ctDNA) by UV???vis absorption, fluorescence, circular dichroism (CD) spectroscopy combined with multivariate curve resolution-alternating least-squares (MCR???ALS) and molecular docking studies. The concentration curves and the pure spectra of compositions (FA, ctDNA and FA???ctDNA complex) were obtained by MCR???ALS approach to verify and monitor the interaction of FA with ctDNA. The groove binding mode between FA and ctDNA was confirmed by the results of melting analysis, viscosity measurements, single-stranded DNA experiments, and competitive studies. The binding constant of FA???ctDNA complex was 4.87?×?10⁴ L mol^?1 at 298?K. The values of enthalpy (ΔH°) and entropy (ΔS°) changes in the interaction were ?16.24?kJ mol^?1 and 35.02?J mol^?1 K^?1, respectively, indicating that the main binding forces were hydrogen bonds and hydrophobic interactions. The result of CD spectra suggested that a decrease in right-handed helicity of ctDNA was induced by FA and the DNA conformational transition from the B-form to the A-form. The results of docking indicated that FA binding with ctDNA in the minor groove. These findings may be conducive to understand the interaction mechanism of FA with ctDNA and the pharmacological effects of FA.

Communicated by Ramaswamy H. Sarma

     

Signal transducer and oxidative stress mediated modulation of phenylpropanoid pathway to enhance rosmarinic acid biosynthesis in fungi elicited whole plant culture of Solenostemon scutellarioides

This study aimed to improve rosmarinic acid (RA) production in the whole plant culture of Solenostemon scutellarioides through elicitation with phytopathogenic fungi. Amongst selected fungi, Aternaria alternata caused significant elevation (p < 0.05–0.01) in RA accumulation (∼1.3–1.6-fold) between 25 and 100 μg l⁻¹. However, elicitation at the dose of 50 μg l⁻¹ has been found to be most effective and intracellular RA content reached almost ∼1.6-fold (p < 0.01) higher in day 7. Therefore, A. alternata (50 μg l⁻¹) was selected for mechanism evaluation. A significant elevation of intercellular jasmonic acid was observed up to day 6 after elicitation with A. alternata (50 μg l⁻¹). A significant increase in tissue H₂O₂ and lipid peroxidation coupled with depletion of antioxidant enzymes superoxide dismutase and catalase indicated augmented oxidative stress associated with biotic interaction. Preceding the elicitor-induced RA accumulation, a notable alteration in the specific activities of biosynthetic enzymes namely PAL and TAT was recorded, while, no significant change in the activities of RAS was observed. HPPR activity was slightly improved in elicited plant. Therefore, it could be concluded that A. alternata elicited the biosynthesis of rosmarinic acid via signal transduction through jasmonic acid coupled with elicitor induced oxidative stress and associated mechanism.     

Development of novel ferulic acid derivatives as potent histone deacetylase inhibitors

Histone deacetylase inhibitors (HDACIs) offer a promising strategy for cancer therapy. The discovery of potent ferulic acid-based HDACIs with hydroxamic acid or 2-aminobenzamide group as zinc binding group was reported. The halogeno-acetanilide was introduced as novel surface recognition moiety (SRM). The majority of title compounds displayed potent HDAC inhibitory activity. In particular, FA6 and FA16 exhibited significant enzymatic inhibitory activities, with IC₅₀ values of 3.94 and 2.82 μM, respectively. Furthermore, these compounds showed moderate antiproliferative activity against a panel of human cancer cells. FA17 displayed promising profile as an antitumor candidate. The results indicated that these ferulic acid derivatives could serve as promising lead compounds for further optimization.     

2-hydroxy-3-phenylpropanoic acid suppressed the growth of Alternaria alternata through damaging cell membrane integrity and modulating reactive oxygen species metabolism

Black spot rot caused by Alternaria alternata is one of the major postharvest disease of apple fruit during logistic. This study evaluated in vitro inhibitory effect of 2-hydroxy-3-phenylpropanoic acid (PLA) at various concentrations on A. alternata and the possible mechanisms involved in its action. Results showed that different concentrations of PLA inhibited conidia germination and mycelial growth of A. alternata in vitro, and 1.0 g L⁻¹ was the lowest effective concentration to suppress A. alternata growth. Moreover, PLA significantly reduced relative conductivity and increased malondialdehyde and soluble protein contents. PLA also increased H₂O₂ and dehydroascorbic acid contents, but reduced ascorbic acid content. Additionally, PLA treatment inhibited catalase, ascorbate peroxidase, monodehydroascorbate acid reductase, dehydroascorbic acid reductase and glutathione reductase activities, whereas promoted superoxide dismutase activity. All these findings suggest that the possible mechanisms involved in the inhibitory effect of PLA on A. alternata included damaging the cell membrane integrity to cause electrolyte leakage and destroying reactive oxygen species balance.     

Direct determination of diene conjugation and deoxyribonucleic acid (DNA) oxidative damage can explain the role of DTPA-Fe2+-O2 complex in a linoleic acid-DNA system

SUMMARY

Exposure of linoleic acid to diethylenetriminepentaacetic acid (DTPA)-Fe²⁺ complexes resulted in fast diene conjugation and peroxidized products which could further react with deoxyribonucleic acid (DNA) to cause DNA oxidative damage. In this paper, we have detected diene conjugation and DNA oxidative damage in a linoleic acid-DNA model system driven by DTPA-Fe²⁺ and found that:

• 1. in air or oxygen-saturated reaction systems, addition of hydrogen peroxide resulted in a decrease in diene conjugation and double-stranded DNA content, but had no obvious effects on the formation of DNA fluorescent products;

• 2. in anoxic conditions, addition of hydrogen peroxide had no effect on the formation of diene conjugation and fluorescent products, but resulted in a decrease of double-stranded DNA content;

• 3. in the presence of DTPA, Fe³⁺ did not stimulate the formation of diene conjugation;

• 4. the formation of diene conjugation and fluorescent products was not inhibited by superoxide dismutase, catalase, sodium benzoate, sodium azide and mannitol.

However, these ‘scavengers’ increased the percentage of double-strand DNA to different degrees. α-tocopherol, but not reduced glutathione (GSH), inhibited the formation of diene conjugates. α-tocopherol and GSH both could reduce the amounts of fluorescent products and DNA strand breaks. Taken together, these data further indicate that chelator-Fe²⁺-O₂ complex, a perferryl-type oxidant, is probably an important initiator of lipid peroxidation in the linoleic acid-DNA system.     

Evaluation of the association of mercury(II) with some dicysteinyl tripeptides

The present study was undertaken to gain insight into the associations of mercury(II) with dicysteinyl tripeptides in buffered media at pH 7.4. We investigated the effects of increasing the distance between cysteinyl residues on mercury(II) associations and complex formations. The peptide–mercury(II) formation constants and their associated thermodynamic parameters in 3-(N-morpholino)propanesulfonic acid (MOPS) buffered solutions were evaluated by isothermal titration calorimetry. Complexes formed in different relative ratios of mercury(II) to cysteinyl peptides in ammonium formate buffered solutions were characterized by LTQ Orbitrap mass spectrometry. The results from these studies show that n-alkyl dicysteinyl peptides (CP 1–4), and an aryl dicysteinyl peptide (CP 5) can serve as effective “double anchors” to accommodate the coordination sites of mercury(II) to form predominantly one-to-one Hg(peptide) complexes. The aryl dicysteinyl peptide (CP 5) also forms the two-to-two Hg₂(peptide)₂ complex. In the presence of excess peptide, Hg(peptide)₂ complexes are also detected. Notably, increasing the distance between the ligating groups or “anchor points” in CP 1–5 does not significantly affect their affinity for mercury(II). However, the enthalpy change (ΔH) values (ΔH₁ ∼ −91 kJ mol⁻¹ and ΔH₂ ∼ −66 kJ mol⁻¹) for complex formation between CP 4 and 5 with mercury(II) are about one and a half times larger than the related values for CP 1, 2 and 3 (ΔH₁ ∼ −66 kJ mol⁻¹ and ΔH₂ ∼ 46 kJ mol⁻¹). The corresponding entropy change (ΔS) values (ΔS₁ ∼ −129 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −116 J K⁻¹ mol⁻¹) of the structurally larger dicysteinyl peptides CP 4 and 5 are less entropically favorable than for CP 1, 2 and 3 (ΔS₁ ∼ −48 J K⁻¹ mol⁻¹ and ΔS₂ ∼ −44 J K⁻¹ mol⁻¹). Generally, these associations result in a decrease in entropy, indicating that these peptide–mercury complexes potentially form highly ordered structures. The results from this study show that dicysteinyl tripeptides are effective in binding mercury(II) and they are promising motifs for the design of multi-cysteinyl peptides for binding more than one mercury(II) ion per peptide.     

Unprecedented derivatization of ferulic acid through selective methoxylation by Aspergillus brasiliensis ATCC 16404

Ferulic acid (FA) is a natural hydroxycinnamic acid widely found in medicinal and edible plants. Several experts have reported the biological potential of FA, including antioxidant and antimicrobial activities. The use of microorganisms in the derivatization of natural products is a useful and advantageous approach to the achievement of high value-added compounds. In order to access chemical derivatives, we conducted the biotransformation of FA by Aspergillus brasiliensis ATCC 16404 for 5 d. In the second day of fermentation, the FA was converted into the new (E)-3-(4-hydroxy-3-methoxyphenyl)-2-methoxyacrylic acid. This is the first time that the extended π-conjugation remained in the chemical structure after the biotransformation of FA. The cytotoxicities of FA and its derivative were evaluated. The biotransformation yielded a derivative less toxic than the parent compound.     

Involvement of polyamines,abscisic acid and anti-oxidative enzymes in adaptation of Blue Panicgrass (Panicum antidotale Retz.) to saline environments

A hydroponic experiment was conducted to assess the possible involvement of polyamines (PAs), abscisic acid (ABA) and anti-oxidative enzymes such as superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) in adaptation of six populations of Panicum antidotale Retz. to selection pressure (soil salinity) of a wide range of habitats. Plants of six populations were collected from six different habitats with ECe ranging from 3.39 to 19.23 dS m⁻¹ and pH from 7.65 to 5.86. Young tillers from 6-month-old plants were transplanted in plastic containers each containing 10 l of half strength Hoagland's nutrient solution alone or with 150 mol m⁻³ NaCl. After 42 days growth, contents of polyamines (Put, Spd and Spm) and ABA, and the activities of anti-oxidative enzymes (SOD, POD and CAT) of all populations generally increased under salt stress. The populations collected from highly saline habitats showed a greater accumulation of polyamines and ABA and the activities of anti-oxidative enzymes as compared to those from mild or non-saline habitats. Moreover, Spm/Spd and Put/(Spd + Spm) ratios generally increased under salt stress. However, the populations from highly saline environments had significantly higher Spm/Spd and Put/(Spd + Spm) ratios as compared to those from mild or non-saline environments. Similarly, the populations adapted to high salinity accumulated less Na⁺ and Cl⁻ in culm and leaves, and showed less decrease in leaf K⁺ and Ca²⁺ under salinity stress. Higher activities of anti-oxidative enzymes and accumulation of polyamines and ABA, and increased Spm/Spd and Put/(Spm + Spd) ratios were found to be highly correlated with the degree of adaptability of Panicum to saline environment.     

Influence of Methionine on Toxicity of Fluoride in the Liver of Rats

Oxidative stress is a common mechanism by which chemical toxicity can occur in the liver. The aim of the studies conducted has been to determine what influence the administration of methionine during intoxication with sodium fluoride may have upon the selected enzymes of the antioxidative system in rat liver. The experiment was carried out on Wistar FL rats (adult females) that, for 35 days, were administered distilled water, NaF, or NaF with methionine (doses: 10 mg NaF/kg bw/day, 10 mg Met/kg bw/day). The influence of administered NaF and Met was examined by analyzing the activity of the antioxidative enzymes: superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and glutathione transferase in the liver. The results suggest that fluoride reduces the efficiency of the enzymatic antioxidative system in the liver. Administration of methionine during intoxication with sodium fluoride does not have an advantageous influence upon the activity of superoxide dismutase, catalase, reductase, and glutathione transferase in the liver. The slight increase of the activity of glutathione peroxidase after administration of methionine may indicate its protective influence upon that enzyme.     

Reactions of the antitumor agent carminic acid and derivatives with DNA

The antitumor agent carminic acid 1a does not bind to DNA but nicks it slowly, more rapidly when reduced in situ, and still more rapidly when prereduced at the quinone moiety. The nicking process requires oxygen and is selectively inhibited by (i) superoxide dismutase, (ii) catalase, and (iii) free radical scavengers indicating the involvement of $\text{O}{}_2{}^{\mathrm{<mtext>?</mtext>}}\text{, H}{}_2\text{O}{}_2$, and OH^., respectively. The intermediacy of OH^. was supported by spin trapping with N-t-butyl-α-phenylnitrone and epr of the radical produced via the carminic acid semiquinone. The single strand scission of DNA by carminic acid requires two adjacent hydroquinone moieties in the chromophore since reduced methyl tetra-O-methylcarminate 1b is without effect although it binds weakly to DNA. Polarographic redox potentials for the reversible (2e, 2H⁺) reduction of 1a and 1b are ?0.736 ± 0.003 V and ?0.56 ± 0.010 V against SCE, respectively. The fact that daunorubicin and adriamycin produce more extensive DNA strand scission than carminic acid under comparable conditions of prereduction and on a molar basis is largely attributed to the assistance of intercalative binding afforded in the case of the anthracyclines.     

Changes in oxidative stress parameters and acid phosphatase activity in the pre-regressing and regressing tail of Indian jumping frog Polypedates maculatus (Anura,Rhacophoridae)

Activities of acid phosphatase (normal and Co²⁺-sensitive), superoxide dismutase and catalase and levels of lipid peroxidation, hydrogen peroxide were compared in the tails of tadpoles of stage III, XVIII, XXI and XXIII, respectively, of the Indian Jumping frog Polypedates maculatus. It is noticed that acid phosphatase activity (normal and Co²⁺-sensitive), and levels of lipid peroxidation and hydrogen peroxide increased during tail regression. There is also an increase in the level of superoxide dismutase and catalase in the regressing tail. A positive correlation between activity of acid phosphatase and lipid peroxidation, hydrogen peroxide and lipid peroxidation, acid phosphatase and hydrogen peroxide was noticed in the tail of tadpoles during different developmental stages, suggesting a critical interaction between reactive oxygen species and lysosomal activity during metamorphosis.     

Release of ferulic acid from corn cobs by alkaline hydrolysis

The process of corn cobs alkaline hydrolysis to produce solutions with high hydroxy-cinnamic acids content was investigated. In particular the attention was focused on the solubilisation of ferulic acid (FA) and related compounds, mainly p-coumaric acid (p-CA). Although these compounds have applications as antioxidants, the purpose of this work was to obtain FA solutions that can be used as feedstock for the biotechnological production of vanillin in future studies. The effects of different concentrations of NaOH (0.2 ≤ C_a ≤ 2.0N) and solid/liquid ratios (0.028 ≤ S/L ≤ 0.168 g/g) on the solubilisation of FA versus time have been investigated at room temperature. Optimal hydrolysis conditions (C_a = 0.5N, S/L = 0.084 g/g after 6 h) ensured the production of hydrolysates with relatively high contents of both FA (1171 ± 34 mg/L) and p-coumaric acid (2156 ± 64 mg/L), which can be used in future studies for the microbial transformation into vanillin.     

Dye degradation,antimicrobial and larvicidal activity of silver nanoparticles biosynthesized from Cleistanthus collinus

The present study aimed in green synthesis and characterization of silver nanoparticles (AgNPs) using the leaves of Cleistanthus collinus. The NPs showed various absorption peaks between 3402 cm⁻¹ and 1063 cm⁻¹. FTIR spectrum revealed the presence of OH group, alkene, aromatic hydrocarbon, aliphatic fluro compound and aliphatic chloro compounds. Scanning electron microscopic analysis revealed the particle size ranged from 30 to 50 nm. The biosynthesized NPs have potent activity against Shigella dysentriae, Staphylococcus aureus and Bacillus subtilis and the zone of inhibition was 21 ± 1, 20 ± 2, 16 ± 2 mm, respectively. Toxicity of the synthesized NPs was tested on green gram (Vigna radiata) seed at various concentrations (20–100%) and germination was induced by NPs treated seeds. Shoot length and root length was higher in NPs treated plant than control plant (p < 0.01). Elevated level of catalase (CAT) and superoxide dismutase (SOD) and about 13% CAT and 7% SOD activity registered than control. Superoxide dismutase activity of root and shoot varied based on the dosage of AgNPs (p < 0.01). Also, the NPs (1%) showed significant larvicidal activity on Aedes aegypti and 100% mortality was achieved after 24 h treatment. The green synthesized NPs reduced methylene blue and 4-nitrophenol significantly (p < 0.01). The colouration of methylene blue and 4-nitrophenol were considerably reduced after 60 min showed the potential of dye degrading ability.     

Anaerobic reduction of ferulic acid to dihydroferulic acid by Wolinella succinogenes from cow rumen

Summary Ferulic acid(FA)-modifying microflora from the rumen of cows were acclimated in an FA-containing medium, in which aromatic compounds (dihydroferulic acid, homovanillic acid, carboxymethylphenol and vanillic acid) and volatile fatty acids (acetic, butyric and isobutyric) were detected by gas chromatography and mass spectrometry. An anaerobic curved bacterium was isolated from the rumen microflora. This bacterium was characterized and identified as Wolinella succinogenes according to the method of Holdeman et al. (1977). It could only reduce FA to dihydroferulic acid in the absence of hydrogen acceptors such as nitrate or fumarate and under strictly anaerobic conditions. FA-reducing ability of the bacterium was inhibited to some extent at FA concentrations greater than 5 mM. The FA was reduced more effectively at pH values of 7.0–7.2 than at 6.8 and the reduction was enhanced by the addition of an Ruminococcus albus' culture supernatant.     

Morphological indicators of salinity stress and their relation with osmolyte associated redox regulation in mango cultivars

Soil salinization is a global issue impeding horticulture production and is approaching an alarming status due to climate change and urbanization. Breeding salt-tolerant rootstock varieties is an ideal strategy to mitigate stress due to salinity in mango and other perennial fruit species. Stress combating strategies employed by seedlings of 7 mango were studied under saline conditions (200 mM NaCl, EC: > 4.0 dSm⁻¹, pH 8.5) in pot experiments. Significantly high accumulation of proline (19.07 µg g⁻¹ FW in Bappakai), glycine betaine (55.11 µg g⁻¹ FW in 13–1), and total sugars (17.33 mg g⁻¹ FW in Kurukkan) were found to be the common mechanism employed by the tolerant cultivars to counter the osmotic stress, under suboptimal conditions. Non-enzymatic antioxidants viz., tannins (17.18 mg g⁻¹), phenols (18.68 mg g⁻¹), and anthocyanins (1.59 mg g⁻¹) were increased in seedling of "13–1", the salt-resistant cultivar from Israel. Reactive oxygen species (ROS) regulation by increased activity of superoxide dismutase and catalase in the two polyembryonic cultivars of Indian origin (Kurukkan and Nekkare) suggests their potential use as rootstocks to combat oxidative stress. The tolerance index of various cultivars was calculated by averaging the scores of morphological stress indicators, and its correlation with studied parameters suggests that salinity resilience is more tightly linked to enhanced catalase accumulation (r² = 0.8361) that is reduced ionic stress. This evidence assign the role of osmotic stress alleviation and redox regulation in salt tolerance mechanism operational in native Indian cultivars, Nekkare and Kurukkan at par with known salt tolerant rootstocks.

     

Synthesis,X-ray crystal structure,DNA/protein binding and cytotoxicity studies of five α-aminophosphonate N-derivatives

Five new α-aminophosphonates are synthesized and characterized by EA, FT-IR, ¹H NMR, ¹³C NMR, ³¹P NMR, ESI-MS and X-ray crystallography. The X-ray analyses reveal that the crystal structures of 1–5 are monoclinic or triclinic system with the space group P 21/c, P − 1, P − 1, P2(1)/c and P − 1, respectively. All P atoms of 1–5 have tetrahedral geometries involving two O-ethyl groups, one C_α atom, and a double bond O atom. The binding interaction of five new α-aminophosphonate N-derivatives (1–5) with calf thymus(CT)-DNA have been investigated by UV–visible and fluorescence emission spectrometry. The apparent binding constant (Kapp) values follows the order: 1 (3.38 × 10⁵ M⁻¹) > 2 (3.04 × 10⁵ M⁻¹) > 4 (2.52 × 10⁵ M⁻¹) > 5 (2.32 × 10⁵ M⁻¹) > 3 (2.10 × 10⁵ M⁻¹), suggesting moderate intercalative binding mode between the compounds and DNA. In addition, fluorescence spectrometry of bovine serum albumin (BSA) with the compounds 1–5 showed that the quenching mechanism might be a static quenching procedure. For the compounds 1–5, the number of binding sites were about one for BSA and the binding constants follow the order: 1 (2.72 × 10⁴ M⁻¹) > 2 (2.27 × 10⁴ M⁻¹) > 4 (2.08 × 10⁴ M⁻¹) > 5 (1.79 × 10⁴ M⁻¹) > 3 (1.17 × 10⁴ M⁻¹). Moreover, the DNA cleavage abilities of 1 exhibit remarkable changes and the in vitro cytotoxicity of 1 on tumor cells lines (MCF-7, HepG2 and HT29) have been examined by MTT and shown antitumor effect on the tested cells.