Highly Oxidized Ellagitannins of Carpinus japonica and Their Oxidation-Reduction Disproportionation.

In the research on ellagitannin metabolism, two unique dehydroellagitannins, carpinins E (1) and F (2), bearing dehydrohexahydroxydiphenoyl (DHHDP) and hydrated biscyclohexenetrione dicarboxyl ester (HBCHT) groups, were isolated from young leaves of Carpinus japonica. Upon heating in H2O or treatment with pH 6 buffer at room temperature, 1 and 2 afforded the reduction product 3, isocarpinin A, with an (R)-hexahydroxydiphenoyl (HHDP) group, suggesting the occurrence of redox disproportionation of the (S)-DHHDP group. This was supported by the increase in production of 3 in the pH 6 buffer solution by coexistence of epigallocatechin-3-O-gallate (15), accompanied by oxidation of 15. In contrast, treatment of 1 and 2 with ascorbic acid yielded 4, carpinin A, with an (S)-HHDP group. Upon heating with ascorbic acid, the HBCHT group was also reduced to an (S)-HHDP group, and 2 was converted to 2,3;4,6-bis(S)-HHDP glucose. In leaves of C. japonica, the tannins 1 and 2 are dominant in young spring leaves, but compounds 3 and 4 become the major components of tannins in mature leaves. These results suggest that, in ellagitannin biosynthesis, oxidative coupling of the two galloyl groups first generates a DHHDP group, and subsequent reduction of DHHDP esters produces HHDP esters.

Production of Ellagitannin Hexahydroxydiphenoyl Ester by Spontaneous Reduction of Dehydrohexa-hydroxydiphenoyl Ester.

Amariin is an ellagitannin with two dehydrohexahydroxydiphenoyl (DHHDP) moieties connecting glucose 2,4- and 3,6-hydroxy groups. This tannin is predominant in the young leaves of Triadica sebifera and Carpinus japonica. However, as the leaves grow, the 3,6-DHHDP is converted to its reduced form, the hexahydroxydiphenoyl (HHDP) group, to generate geraniin, a predominant ellagitannin of the matured leaves. The purified amariin is unstable in aqueous solution, and the 3,6-(R)-DHHDP is spontaneously degraded to give HHDP, whereas 2,4-(R)-DHHDP is stable. The driving force of the selective reduction of the 3,6-DHHDP of amariin is shown to be the conformational change of glucose from O,3B to 1C4. Heating geraniin with pyridine affords 2,4-(R)-DHHDP reduction products. Furthermore, the acid hydrolysis of geraniin yields two equivalents of ellagic acid. Although the reaction mechanism is still ambiguous, these results propose an alternative biosynthetic route of the ellagitannin HHDP groups.

Impact of the combination of sourdough fermentation and hop extract addition on baking properties, antioxidant capacity and phenolics bioaccessibility of rice bran-enhanced bread.

The effect of hop sourdough fermentation combined with rice bran (RB) addition was investigated for improving technological, bioactive and sensory properties of wheat bread by optimizing the quantities of RB, yeast, sourdough and hop extract contents based on response surface methodology. The bread produced according to the optimized formula was compared with control bread, sourdough or RB-enhanced bread with regard to quality attributes, bioaccessible phenolics and antioxidant activity. Sourdough fermentation combined with RB flour increased volume of loaves and consumer acceptance with no significant variations in crumb moisture content and crumb firmness during storage. RB darkened crumb while no effects on colour were observed for sourdough bread. Sourdough technology allowed increasing phenolic content of RB bread with no significant differences in its antioxidant activity. The bioaccessible ferulic and sinapic acids, the main phenolics released in vitro digestion of optimized bread were lower compared to those of control bread.

Anti-skin ageing effects of phenolic compounds from Carpinus tschonoskii.

Carpinus tschonoskii (CT) is distributed through in southern regions of South Korea. Six known compounds, including three ellagitannins, one gallotannin, and two flavonoids were isolated from CT. In conclusion, the tannins, especially ellagitannins, and CT extract showed potent anti-oxidative, anti-inflammatory and anti-skin ageing activities.

On the use of dorsiventral reflectance asymmetry of hornbeam (Carpinus betulus L.) leaves in air pollution estimation.

This study examines the role of dorsiventral leaf measurements in reflectance-based air quality estimation. The dorsiventral asymmetry is used to describe the difference between the upper (adaxial) and lower (abaxial) leaf side. Spectral characteristics of dorsiventral asymmetry and both adaxial and abaxial leaf reflectance are investigated for a typical dicotyledonous species Carpinus betulus used in an urban environment. The link with traffic-related air pollution is established and the potential for monitoring of air quality is evaluated. We conclude that dorsiventral reflectance asymmetry is a factor that should not be ignored in canopy measurements and modeling. On the other hand, the benefits of dorsiventral asymmetry indices as a tool for reflectance-based air quality seem limited.

Hyperspectral leaf reflectance of Carpinus betulus L. saplings for urban air quality estimation.

In urban areas, the demand for local assessment of air quality is high. The existing monitoring stations cannot fulfill the needs. This study assesses the potential of hyperspectral tree leaf reflectance for monitoring traffic related air pollution. Hereto, 29 Carpinus betulus saplings were exposed to an environment with either high or low traffic intensity. The local air quality was estimated by leaf saturation isothermal remanent magnetization (SIRM). The VIS-NIR leaf reflectance spectrum (350-2500 nm) was measured using a handheld AgriSpec spectroradiometer (ASD Inc.). Secondary, leaf chlorophyll content index (CCI), specific leaf area (SLA) and water content (WC) were determined. To gain insight in the link between leaf reflectance and air quality, the correlation between SIRM and several spectral features was determined. The spectral features that were tested are plain reflectance values, derivative of reflectance, two-band indices using the NDVI formula and PCA components. Spectral reflectance for wavelength bands in the red and short wave IR around the red edge, were correlated to SIRM with Pearson correlations of up to R = -0.85 (R2 = 0.72). Based on the spectral features and combinations thereof, binomial logistic regression models were trained to classify trees into high or low traffic pollution exposure, with classification accuracies up to 90%. It can be concluded that hyperspectral reflectance of C. betulus leaves can be used to detect different levels of air pollution within an urban environment.

Pyrolysis of hornbeam (Carpinus betulus L.) sawdust: Characterization of bio-oil and bio-char.

Slow pyrolysis of hornbeam (Carpinus betulus L.) sawdust was performed to produce bio-oil and bio-char. The operational variables were as follows: pyrolysis temperature (400-600°C), heating rate (10-50°Cmin-1) and nitrogen flow rate (50-150cm3min-1). Physicochemical and thermogravimetric characterizations of hornbeam sawdust were performed. The characteristics of bio-oil and bio-char were analyzed on the basis of various spectroscopic and chromatographic techniques such as FTIR, GC-MS, 1H NMR, SEM, BET. Higher heating value, density and kinematic viscosity of the bio-oil with maximum yield of 35.28% were 23.22MJkg-1, 1289kgm-3 and 0.6mm2s-1, respectively. The bio-oil with relatively high fuel potential can be obtained from the pyrolysis of the hornbeam sawdust and the bio-char with a calorific value of 32.88MJkg-1 is a promising candidate for solid fuel applications that also contributes to the preservation of the environment.

Seasonal variation of pheophorbide a and flavonoid in different organs of two Carpinus species and its correlation with immunosuppressive activity.

The genus Carpinus of Betulaceae is the most widely distributed in the European landscape. This study reports a comparative study based on the pheophorbide a and flavonoid content from the two main species of the genus Carpinus, Carpinus betulus and Carpinus turczaninowii, respectively, in Nanjing, China. The pheophorbide a and flavonoid content depends on the organ, species, and season. HPLC analysis showed that the pheophorbide a and flavonoid levels were the highest in May and June, respectively, from the leaves of C. betulus 'Fastigiata.' In contrast, the content of pheophorbide a and flavonoid in the stems of C. betulus 'Fastigiata' or in other species was low. The immunosuppressive effects of the ethyl acetate extracts and methanol extracts from the two Carpinus species were also evaluated. The ethyl acetate extracts of C. betulus 'Fastigiata' in May and the methanol extracts of C. betulus 'Fastigiata' in June showed better immunosuppressive activity than in other seasons, which coincided with the content of pheophorbide a and flavonoid, respectively. Our findings indicated that C. betulus 'Fastigiata' can serve as a medicinal plant against inflammation because of its pheophorbide a and flavonoid content.

Inhibitory effects of Carpinus tschonoskii leaves extract on CpG-stimulated pro-inflammatory cytokine production in murine bone marrow-derived macrophages and dendritic cells.

This report describes the anti-inflammatory effects of MeOH extract from leaves of Carpinus tschonoskii (CE) on primary bone marrow-derived macrophage (BMDMs) and dendritic cells (BMDCs). Primary BMDMs and BMDCs were used for pro-inflammatory cytokine production and Western blot analysis. Human embryonic kidney cell line 293 T (HEK293 T) was used to access NF-κB activity. In all cases, CpG DNA was used to stimulate the cells. The CE (0-150 µg/ml) was treated to BMDMs, BMDCs, and HEK293T cells. CE pre-treatment in CpG-stimulated BMDMs and BMDCs showed a dose-dependent inhibitory effect on pro-inflammatory cytokine (e.g., IL-12 p40, IL-6, and TNF-α) production as compared to non-treated controls. The CE pre-treatment had no significant inhibition on mitogen-activated protein kinases (MAPKs) phosphorylation but strongly inhibited IκBα degradation. In NF-κB reporter gene assay, the CE pre-treatment inhibited NF-κB-dependent luciferase activity in a dose-dependent manner. Taken together, these data suggest that CE has significant inhibitory effect on pro-inflammatory cytokine production and warrant further studies concerning potentials of CE for medicinal uses.

Potential anticancer activity of young Carpinus betulus leaves.

As part of our continuing research for anticancer compounds from the Walloon Region forest, EtOAc extract from Carpinus betulus leaves was phytochemically studied, leading to the bioguided isolation of pheophorbide a, which is responsible of anticancer properties of C. betulus young leaves. This compound was identified using nuclear magnetic resonance and mass spectrophotometric data and comparison with a commercial standard. Evaluation of the growth inhibitory activities of pheophorbide a using MTT colorimetric assay and phase-contrast microscopy in various human cancer cell lines confirmed the photoactivable properties of this compound. Our research showed, for the first time, the presence of pheophorbide a, a chlorophyll derived compound, which we quantified in high quantities in young leaves of C. betulus. This is in contrast with the literature which generally describes pheophorbide a as a catabolic product of chlorophyll, then preferentially present in old leaves.

Naturally occurring diarylheptanoids.

Diarylheptanoids, natural products with a 1,7-diphenylheptane structural skeleton, are mainly distributed in the roots, rhizomes and bark of Alpinia, Zingiber, Curcuma and Alnus species. They have become of interest in natural product research over the past twenty years because of their remarkable anti-cancer, anti-emetic, estrogenic, anti-microbial and anti-oxidant activity. This paper compiles all 307 naturally occurring diarylheptanoids from 46 plants as reported in 137 references with their distributions, physiological activities and 13C-NMR spectral data.

Possible fungistatic implications of betulin presence in betulaceae plants and their hymenochaetaceae parasitic fungi.

Betulin and its derivatives (especially betulinic acid) are known to possess very interesting prospects for their application in medicine, cosmetics and as bioactive agents in pharmaceutical industry. Usually betulin is obtained by extraction from the outer layer of a birch bark. In this work we describe a simple method of betulin isolation from bark of various species of Betulaceae trees and parasitic Hymenochaetaceae fungi associated with these trees. The composition of the extracts was studied by GC-MS, whereas the structures of the isolated compounds were confirmed by FTIR and 1H NMR. Additionally, the significant fungistatic activity of betulin towards some filamentous fungi was determined. This activity was found to be strongly dependent on the formulation of this triterpene. A betulin-trimyristin emulsion, in which nutmeg fat acts as emulsifier and lipophilic carrier, inhibited the fungal growth even in micromolar concentrations--its EC50 values were established in the range of 15 up to 50 microM depending on the sensitivity of the fungal strain. Considering the lack of fungistatic effect of betulin applied alone, the application of ultrasonic emulsification with the natural plant fat trimyristin appeared to be a new method of antifungal bioassay of water-insoluble substances, such as betulin.

Hydrolysable tannins depress cardiac papillary muscle contraction and propranolol-induced negative inotropism.

We studied effects of hydrolysable tannins on rat papillary muscle contractions induced by propranolol. The developed force of papillary muscle was measured isometrically with an inductive force transducer. The IC(50) values for digallic acid, gallic acid, germin D, praecoxin A and 1-desgalloyl rugosin F were 4.2 x 10(-5) M, 1.3 x 10(-4) M, 1,4 x 10(-4) M, 1.5 x 10(-4) and 1.7 x 10(-4) M, respectively. Incubation with tannins significantly attenuated the propranolol-induced negative inotropic contractile response. These results show that hydrolysable tannins depress muscle contractions and potentiate the activities of ß-adrenergic blocker.

Synthesis of a molecularly imprinted polymer for the solid-phase extraction of betulin and betulinic acid from plane bark.

INTRODUCTION: Plant extracts are usually complex mixtures of various polarity compounds and their study often includes a purification step, such as solid-phase extraction (SPE), to isolate interest compounds prior analytical investigations. Molecularly imprinted polymers (MIPs) are a new promising type of SPE material which offer tailor-made selectivity for the extraction of trace active components in complex matrices. Numerous specific cavities that are sterically and chemically complementary of the target molecules, are formed in imprinted polymers. A molecularly imprinted polymer (MIP) was synthesised in order to trap a specific class of triterpene, including betulin and betulinic acid from a methanolic extract of plane bark. METHODOLOGY: Imprinted polymers were synthesised by thermal polymerisation of betulin as template, methacrylic acid (MAA) or acrylamide (AA) as functional monomer, ethylene glycol dimethacrylate as crosslinking agent and chloroform as porogen. Afterwards, MAA- and AA-MIPs were compared with their non-imprinted polymers (NIPs) in order to assess the selectivity vs betulin and its derivatives. Recovered triterpenes were analysed by HPLC during MIP-SPE protocol. RESULTS: After SPE optimisation, the MAA-imprinted polymer exhibited highest selectivity and recovery (better than 70%) for betulin and best affinity for its structural analogues. Thus, a selective washing step (chloroform, acetonitrile) removed unwanted matrix compounds (fatty acids) from the SPE cartridge. The elution solvent was methanol. Finally, the MAA-MIP was applied to fractionate a plane bark methanolic extract containing betulin and betulinic acid. CONCLUSION: This study demonstrated the possibility of direct extraction of betulin and its structural analogues from plant extracts by MIP technology.

Physical and mechanical properties of European Hophornbeam (Ostrya carpinifolia Scop.) wood.

The main purpose of the study was to determine some physical and mechanical properties of European Hophornbeam (Ostrya carpinifolia Scop.) wood of which is not any detailed previous research in the literature. The sample trees harvested from a mixed beech-oak-hornbeam stand in the Zonguldak Forest Enterprises, north western part of Turkey. Conventional methods followed and the test procedures were performed on small clean specimens. European Hophornbeam wood's air dry and over dry densities were determined as 890 and 853 kg m(-3); density value in volume (basic density) was determined as 671 kg m(-3); volumetric shrinkage and swelling were determined as 23.02% and 24.94%; Fiber saturation point (FSP) was found 34.21%; MOR, MOE, compression strength parallel to grain, impact bending, tensile strength parallel and perpendicular to grain, and Janka hardness values (parallel and perpendicular to grain) were determined as 131.50 N/mm2, 11501.06 N/mm2, 66.94 N/mm2, 18.66 N/mm2, 105.49 N/mm2, 7.11 N/mm2, 6.89 and 5.63 kN, respectively.

New diarylheptanoids from the stems of Carpinus cordata.

Two new diarylheptanoids, carpinontriols A (1) and B (2), were isolated from the stems of Carpinus cordata, along with the known diarylheptanoid, casuarinondiol (3), and five known compounds, (+)-catechin (4), methyl gallate (5), methyl gallate 3-O-beta-D-glucopyranoside (6), methyl gallate 4-O-beta-D-glucopyranoside (7), and methyl gallate 3-O-beta-D-(6'-O-galloyl)-glucopyranoside (8). The structures of 1 and 2 were elucidated by spectral methods. Among the isolated compounds, compounds 4-6 and 8 showed radical-scavenging activity in the DPPH assay.