Polyphenolic compounds, antioxidant capacity, and quinone reductase activity of an aqueous extract of Ardisia compressa in comparison to mate (Ilex paraguariensis) and green (Camellia sinensis) teas

Aqueous extracts of the leaves of Ardisia compressa (AC) have been used in folk medicine to treat various liver disorders including liver cancer. The objective of this study was to partially characterize and determine the total polyphenol content, antioxidant capacity, and quinone reductase activity of A. compressa tea in comparison to mate (Ilex paraguariensis, MT) and green (Camellia sinensis,GT) teas. Total polyphenol content, antioxidant capacity, and phase II enzyme induction capacity were measured by the modified Folin-Ciocalteu, ORAC, and quinone reductase (QR) assays, respectively. The major polyphenols in AC were not catechins. HPLC retention times and standard spikes of AC indicated the presence of gallic acid, epicatechin gallate, ardisin and kaempferol. Using catechin as standard, the total polyphenol value of AC (36.8 +/- 1.1 mg/mg DL) was significantly lower than GT (137.2 +/- 5.8 mg equivalent of (+)-catechin/mg dried leaves, DL) and MT (82.1 +/- 3.8 mg/mg DL) (P < 0.001). Antioxidant capacity (AC, 333; GT, 1346; MT, 1239 mmol Trolox equivalents/g DL) correlated with total polyphenol values (r(2) = 0.86, P < 0.01). AC (4.5-12.5 microg/mL) induced QR enzyme, in Hepa1c1c7 cells, up to 15%. MT and GT showed no induction at the concentrations tested (0.5-10.5 and 0.5-12.5 mg/mL, respectively). These results suggest that AC has a different mechanism of protection against cytotoxicity that is not related to its antioxidant capacity. Further studies are needed to determine such mechanisms and to explore its potential as a chemopreventive or therapeutic agent.     

Characterization of the constituents and antioxidant activity of Brazilian green tea (Camellia sinensis var. assamica IAC-259 cultivar) extracts

Freeze-dried extracts from Camellia sinensis var. assamica IAC-259 cultivar named Brazilian green tea were prepared by hot water and ultrasound-assisted extractions using leaves harvested in spring and summer. Their caffeine and catechin contents were measured by high performance liquid chromatography-diode array detector. The antioxidant activity of the major green tea compounds and Brazilian green tea extracts was evaluated using a 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay. The levels of caffeine were higher in the summer samples (p < 0.05); otherwise, there were no significant variations related to the catechin contents between spring and summer samples. The sonication method using water/acetone as solvent had a high efficiency to extract not only epigallocatechin gallate but also epicatechin gallate (p < 0.05). Antioxidant activities of the Brazilian green tea extracts were not significantly different among seasons and extraction systems. The antioxidant data (IC50) of the Brazilian green tea extracts showed a significant correlation with their epigallocatechin gallate and epicatechin gallate contents (p < 0.05).     

Inhibition of pathogenic bacterial adhesion by acidic polysaccharide from green tea (Camellia sinensis)

An acidic polysaccharide CS-F2 from Camellia sinensis was examined to characterize its anti-adhesive effects against pathogenic bacteria, most notably Helicobacter pylori, Propionibacterium acnes, and Staphylococcus aureus. CS-F2 showed marked inhibitory activity against the pathogen-mediated hemagglutination with a minimum inhibitory concentration (MIC) between 0.01 and 0.1 mg/mL, which is lower than the previously reported MIC values for Panax ginseng and Artemisia capillaris. The inhibitory effects of CS-F2 on the adhesion of H. pylori to AGS adenocarcinoma gastric epithelial cells, or P. acnes and S. aureus to NIH 3T3 fibroblast cells, were further assessed resulting in MIC values between 0.063 and 0.13 mg/mL. Importantly, CS-F2 showed no inhibitory effects against Lactobacillus acidophilus, Escherichia coli, or Staphylococcus epidermidis. Our results suggest that CS-F2, which is a pectin-type polysaccharide with a molecular weight of approximately 8.0 x 10(4) Da, may exert a selective anti-adhesive effect against certain pathogenic bacteria, while exerting no effects against beneficial and commensal bacteria.     

Immunostimulating activity of a crude polysaccharide derived from green tea (Camellia sinensis) extract

Green tea extract is well-known to reduce the risk of a variety of diseases. Here, we investigated the immunostimulating activity of tea polysaccharide (TPS), one of the main components in green tea extract. The water extracts from mature or immature tea leaves were precipitated by using ethanol at room temperature. The sediment was washed with ethanol and acetone alternately and then dried. We used the phagocytic activity of macrophage-like cells as an indicator of immune function activation. Chemical components were analyzed by HPLC. The immunostimulating activity of TPS from immature leaves extract was higher than that of TPS from mature leaves, and its activities were dependent on the content of strictinin in the leaf extract. Futhermore, a mixture of catechin and TPS that removed polyphenols did not increase the immunostimulating activity. These results suggest that the catechin-polysaccharide complex is a very important molecule in the immunomodulating activity of tea extracts.     

Total polyphenol content and antioxidant capacity of commercially available tea (Camellia sinensis) in Argentina

Tea, Camellia sinensis (L.) O. Kuntze (Theaceae) is cultivated in Argentina in the northeastern region (provinces of Misiones and Corrientes), between 26 degrees and 28 degrees south latitude, the southernmost area of the world where tea is cultivated. The objective of this work was to determine the total polyphenol content and the in vitro antioxidant capacity of green and black tea cultivated and industrialized in Argentina. Twelve samples of eight brands were analyzed. The total polyphenol content was determined according to the International Organization for Standardization method (ISO) 14502-1 for the determination of substances characteristic of green and black tea. The antioxidant capacity was determined by the ferric thiocyanate method (FTC) and the 1,1-diphenyl-2-picrylhydrazyl (DPPH) free-radical scavenging assay. Green tea showed a higher polyphenol content than black tea. The total polyphenol concentration in green tea was found to vary from 21.02 +/- 1.54 to 14.32 +/- 0.45% of gallic acid equivalents (GAE), whereas in black tea, the polyphenol content ranged from 17.62 +/- 0.42 to 8.42 +/- 0.55% of GAE (P < 0.05). A similar profile was observed for the antioxidant capacity determined by both methods. The antioxidant activities were well correlated with the total polyphenol content (r (2) = 0.9935 for the ferric thiocyanate method and r (2) = 0.9141 for the 1,1-diphenyl-2-picrylhydrazyl free-radical scavenging assay). This is the first systematic screening for the quantification of polyphenols and antioxidant activity in tea commercialized in Argentine markets. The results obtained herein allow one to conclude that Argentine tea is of very good quality when compared to teas from other sources.     

Pigments in green tea leaves (Camellia sinensis) suppress transformation of the aryl hydrocarbon receptor induced by dioxin

Environmental contaminants such as dioxins enter the body mainly through diet and cause various toxicities through transformation of the aryl hydrocarbon receptor (AhR). We previously reported that certain natural flavonoids at the dietary level suppress the AhR transformation induced by 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD). In this study, we identified lutein and chlorophyll a and b from green tea leaves as the novel antagonists for AhR. These active compounds suppressed AhR transformation dose-dependently with the 50% inhibitory concentration (IC(50)) values against 0.1 nM TCDD-induced AhR transformation at 3.2, 5.0, and 5.9 microM, respectively. (-)-Epigallocatechin gallate, which is the most abundant flavonoid in green tea leaves, also showed stronger suppressive effects than did other major tea components, with the IC(50) value of 1.7 microM. Thus, these pigments of green tea leaves have the potential to protect from dioxin toxicity through the suppression of AhR transformation.     

Metabolite profiling using (1)H NMR spectroscopy for quality assessment of green tea, Camellia sinensis (L.)

A set of 191 green teas from different countries was collected and analyzed by (1)H NMR. It was proposed to establish if the teas could be discriminated according to the country of origin or with respect to quality. Both principal component analysis (PCA) and cluster analysis were applied to the data. Some separation of Chinese and non-Chinese teas was observed. The present results did not allow allocation of samples to individual countries, but cluster analysis suggested that it might be possible with an augmented sample set. The PCA did show a separation between the Longjing type (highest quality Chinese tea) and most other Chinese teas and indicated some metabolites that could be responsible for the difference. Longjing teas showed higher levels of theanine, gallic acid, caffeine, epigallocatechin gallate, and epicatechin gallate and lower levels of epigallocatechin when compared with other teas. These compounds have been mentioned previously in connection with quality, but it was also shown that higher levels of theogallin (5-galloyl quinic acid), theobromine, 2-O-(beta-l-arabinopyranosyl)-myo-inositol and some minor sugar-containing compounds were found in Longjing teas while higher levels of fatty acids and sucrose were found in the other teas. These new markers could prove to be useful for the authentication of bulk tea.     

The use of catechins as biochemical markers in diversity studies of tea (Camellia sinensis)

The concentrations of catechins in a Kenyan tea germplasm collection of 102 accessions were determined by HPLC. Total green leaf catechin concentrations and the ratio of dihydroxylated to trihydroxylated catechins were used to establish genetic differentiation in the germplasm. Upon multivariate analysis, accumulation of the various catechins separated the tea clones into 3 major and 5 minor groups according to their phylogenetic origins. The Cambod teas had the highest ratio (7:10) followed by China teas (3:5) while Assam teas had the lowest ratio (1:4). This biochemical differentiation indicates that there is potential for broadening the genetic base of the mainly Assam teas in Kenya (90%) with the putative China and Cambod teas.     

Comparison of antioxidant and antimicrobial activities of tilia (Tilia argentea Desf ex DC), sage (Salvia triloba l.), and black tea (Camellia sinensis) extracts

The antioxidant activity of the water extract of Tilia argentea Desf ex DC was determined by the thiocyanate method. The antioxidant activity of the water extract increased with the increasing amount of lyophilized extract (50-400 microg) added into the linoleic acid emulsion. Statistically significant effect was determined in 100 microg and higher amounts. Antioxidant activities of water extracts of tilia (Tilia argentea Desf ex DC), sage (Salvia triloba L.), and two Turkish black teas commercially called Rize tea and young shoot tea (Camellia sinensis) were compared. For comparison studies, 100 microg portions of extracts were added into test samples. All samples were able to show statistically significant antioxidant effect. Both of the tea extracts showed highest antioxidant activities, nevertheless, differences between tilia and sage and tilia and tea were not statistically significant (for both cases p > 0.05). Like antioxidant activity, the reducing power of water extract of Tilia argentea Desf ex DC was also concentration dependent. Even in the presence of 50 microg of extract, the reducing power was significantly higher than that of the control (p < 0.05) in which there was no extract. Unlike antioxidant activity, the highest reducing power activity was shown by sage extract. Among the tea extracts, young shoot extract was the most effective one, however, it had significantly lower activity than sage (p < 0.05). Although tea flower had the lowest reducing power activity, it was higher than that of tilia. But this difference was not statistically significant (p > 0.05). From these results, we could suggest that although the reducing power of a substance may be an indicator of its potential antioxidant activity, there may not always be a linear correlation between these two activities. In addition, antimicrobial activities of each of the above extracts were studied by disk diffusion methods on different test microorganisms. None of the extracts showed antibacterial activity on the studied microorganisms.     

The coupling effect of water and fertilizer on the growth of tea plants [Camellia sinensis (L.) O. Kuntz]

Water and fertilizer are the two main factors promoting the rapid growth of tea plants (Camellia sinensis). Using two-year-old rooted cutting of Wuniuzao (Camellia sinensis) as experimental materials, “311-B” D-saturation optimization design was carried out using 5, 4, and 4 levels of irrigation regimes, fertilizer nitrogen, and fertilizer phosphorus, with six replications. The raw date obtained was used of quadratic regression fitting for the plant height, new-tip length and dry matter weight. The coupling effects of these three factors [nitrogen (N0, water, and phosphorus (P)] on dry matter weight were significant. The effect order of single factor on dry matter weight was N>water>P, while the interactive effect order on the dry matter weight was N and water>P and water>N and P. An optimal combination with 0.485 g·pot^?1 N, 0.274 g·pot^?1 P, and once every 4-5 days of watering was proposed for the highest tea dry matter yield.     

Effect of yerba mate (Ilex paraguariensis) tea on topoisomerase inhibition and oral carcinoma cell proliferation

Tea flavonoids have antitopoisomerase activity and can inhibit cell proliferation. The objectives of this study were to determine the phenolic content of yerba mate tea products (MT) (Ilex paraguariensis) and evaluate their capacity to inhibit topoisomerase I (Topo I) and II (Topo II) activities and oral carcinoma cell proliferation. Total polyphenols of aqueous extracts of dried MT leaves were measured by the Folin-Ciocalteau assay, using chlorogenic (CH) and gallic (GA) acids as standards. Topoisomerase inhibition was determined by a clone-forming assay, which uses yeast (Saccharomyces cerevisiae) strains as a model. Controls included dimethyl sulfoxide (1.66%); camptothecin (50 microg/mL), a Topo I inhibitor; and amsacrine (100 microg/mL), a Topo II inhibitor. Cytotoxicity studies were conducted using a nontumorigenic human keratinocyte cell line HaCaT and two human squamous cancer cell lines (SCC-61 and OSCC-3). MT was found to be a rich source of phenolic compounds. Total polyphenol content of various commercially available traditional MT products ranged from 236 to 490 mg equiv of CH/g of dry leaves. Such levels were significantly different among products depending on their origin (P < 0.001). Higher anti-topoisomerase II activity was observed against JN394t(2-4) strain for Nobleza Gaucha MT (IC50 = 0.43 microg equiv of CH) in comparison to GA (IC50 = 112 mM) and CH (IC50 > 1500 mM). MT showed catalytic anti-topoisomerase activity against Topo II but not against Topo I. In addititon, MT exhibited dose-dependent cytotoxicity against all squamous cell lines tested. In comparison to premalignant cell line HaCaT [28 microg equiv of (+)-catechin mL(-1)], the cell line SCC-61 [21 microg equiv of (+)-catechin mL(-1)] was the most sensitive to MT, resulting in 50% inhibition of net cell growth. It is concluded that MT is rich in phenolic constituents and can also inhibit oral cancer proliferation. The effect on cancer cell proliferation may be mediated via inhibition of topoisomerase II. The lack of correlation between polyphenol content and the inhibition of topoisomerases suggests that the effect of MT on topoisomerase inhibition may be due to other still unidentified biologically active phytochemicals constituents.     

Inhibition of gastrointestinal lipolysis by green tea, coffee, and gomchui (Ligularia fischeri) tea polyphenols during simulated digestion

Green tea, coffee, and gomchui (Ligularia fischeri) tea, which are rich in polyphenols, may exhibit antiobesity effects by inhibiting pancreatic lipase. However, the bioavailability of some polyphenols is poor due to either degradation or absorption difficulties in the gastrointestinal tract, thus making their beneficial effects doubtful. This study was conducted to evaluate the inhibitory effect of three beverages on lipolysis and the contribution of their major polyphenols during simulated digestion. During simulated digestion, gomchui tea was the most potent at inhibiting gastrointestinal lipolysis, whereas green tea was the least potent. The strongest lipase inhibitor among purified major polyphenols was a green tea polyphenol, (-)-epigallocatechin gallate (EGCG, IC(50) = 1.8 ± 0.57 μM), followed by di-O-caffeoylquinic acid isomers (DCQA, IC(50) from 12.7 ± 4.5 to 40.4 ± 2.3 μM), which are gomchui tea polyphenols. However, the stability of DCQA was greater than that of EGCG when subjected to simulated digestion. Taken together, gomchui tea, which has DCQA as the major polyphenol, showed stronger lipolysis inhibitory activity during simulated digestion compared to both green tea and coffee.     

Influence of agronomic variables on the macronutrient and micronutrient contents and thermal behavior of mate tea leaves (Ilex paraguariensis)

The influence of agronomic variables (light intensity, age of leaves, and fertilization type) on the content of macronutrients and micronutrients (potassium, calcium, sodium, magnesium, manganese, iron, zinc, and copper) of tea leaves was assessed by acid digestion, followed by flame atomic absorption spectrometry (FAAS). The thermal behavior of mate tea leaves (Ilex paraguariensis) was also studied in this work. Samples of mate (Ilex paraguariensis) were collected in an experiment conducted under agronomic control at Erva-Mate Bar?o Commerce and Industry LTD (Brazil). The results showed that the mineral content in mate is affected by the agronomic variables investigated. In general, the content of mineral compounds analyzed is higher for younger leaves and for plants cultivated in shadow. Thermal analysis of samples indicated a similar behavior, with three typical steps of decomposition: loss of water, degradation of low-molecular weight compounds, and degradation of residual materials.     

钙[Ca(NO_3)_2和CaO]对茶树氟吸收的影响

通过水培和土壤盆栽试验,研究了Ca对茶树F吸收的影响。结果显示,在含F溶液中添加Ca显著降低茶树对F的吸收。在土壤盆栽试验中也得到了类似结果,在2种土壤F含量条件下,加入Ca(NO3)2后显著降低茶树成熟叶和新梢F含量。同样土壤加入CaO后,显著降低茶树对F的吸收。土壤分析表明加入Ca(NO3)2后并没有显著改变土壤水溶性F含量,而加入CaO后还增加了土壤水溶性F含量。因此,Ca减少F吸收可能并不单纯是因为形成CaF2后降低土壤F的有效性。     

Reddish coloration of Chinese quince (Pseudocydonia sinensis) procyanidins during heat treatment and effect on antioxidant and antiinfluenza viral activities

To investigate the effects of processing Chinese quince fruit on the denaturation of phenolics and their food functions, fruit phenolic extracts were heated together with organic acid for up to 12 h. Chinese quince phenolic (mostly procyanidins) solution subjected to heat treatment changed from almost colorless, pale yellow, to a reddish color. Before heat treatment, the absorption spectra of polymeric procyanidins were observed only around 280 nm; after heat treatment, absorption occurred between 400 and 600 nm, which is related to the reddish color appearance. Thioacidolysis of denatured reddish phenolics showed that (-)-epicatechin subunits decreased during heat treatment and, in contrast, cyanidin increased. In addition, novel substances that could not be degraded by thioacidolysis were formed. Meanwhile, antioxidant activities, assessed by linoleic acid peroxidation, 1,1-diphenyl-2-picrylhydrazyl (DPPH), Folin-Ciocalteu, and FRAP methods, increased during heat treatment. The antiinfluenza viral activity of denatured reddish phenolics was inferior to that of intact fruit phenolics; however, they retained moderate activity. These results indicate that red coloration of fruit products of Chinese quince was mainly due to the spectral (i.e. structural) changes of procyanidins accompanied with formation of cyanidin. Increasing the length of heat treatment increased the antioxidant capacity of phenolics, and the resultant reddish phenolics retained moderate antiinfluenza viral activity.     

茶树新梢cDNA克隆测序和表达序列标签(ESTs)特性分析

采用定向克隆法，构建了茶树(Camellia sinensis(L．)O．Kuntze)龙井43和安吉白茶等2个品种新梢cDNA文库。对龙井43cDNA文库共4320个克隆的序列进行了测定，获得有用序列2963个，其中空载体和去除载体后序列短于150bp有416个，重复的有863个，首批共获得1684个茶树ESTs。绝大部分ESTs的长度在300～700bp之间，平均为478bp。通过与NCBI等核酸数据库进行比对、查询和注释，获得已知功能基因或具推测功能的基因130多个(607个ESTs)，新的表达基因1077个，证明ESTs测序分析是一个发现和鉴定茶树功能基因的高效快速新途径。     

Model studies on the influence of matrix type and storage environment on the stability of a model aroma mixture during storage

The objective of this study was to investigate the effect of oxygen in the storage atmosphere on the degradation of model compounds when present in water or a medium chain triglyceride (MCT) matrix. A model aroma compound mixture was prepared in oil (MCT) or water, and it was then stored under either an ambient air or argon atmosphere containing respectively ca. 20% and <0.5% residual oxygen. Samples were analyzed by SPME-GC/MS to determine the relative stability over time of different classes of aroma compounds. The low-oxygen atmosphere appeared to have a significant protective effect on sulfur compounds, aldehydes, and ketones in oil but a detrimental influence on pyrroles. Data showed little influence of the atmosphere for these compounds in water. In addition, the type of matrix had a significant effect (P < 0.05) on the stability of aldehydic, ester, and pyrrole compounds. These compounds were more stable in MCT than in water.     

The apoptotic effect of green tea (-)-epigallocatechin gallate on 3T3-L1 preadipocytes depends on the Cdk2 pathway

This study was designed to investigate the effect of green tea catechins, especially (-)-epigallocatechin gallate (EGCG), on the apoptosis of 3T3-L1 preadipocytes. Preadipocyte apoptosis as indicated by formation of DNA fragments was induced by EGCG in dose-dependent manners. While EGCG was demonstrated to decrease Cdk2 expression and activity and increase caspase-3 activity, overexpression of Cdk2 and treatment with the caspase-3 inhibitor respectively prevented preadipocytes from induction of DNA fragmentation and caspase-3 activity by doses of 100-400 muM of EGCG. This suggests the Cdk2- and caspase-3-dependent apoptotic effects of EGCG. Moreover, EGCG was more effective than EC, ECG, and EGC in changing the apoptotic signals. Results of this study may relate to the mechanism by which EGCG modulates body weight.     

Impact of blanching and packaging atmosphere on the formation of aroma compounds during long-term frozen storage of leek (Allium ampeloprasum Var. Bulga) slices.

The content of aroma compounds and the catalytic activity of lipoxygenase (LOX), alliinase, hydroperoxide lyase (HPL), and alcohol dehydrogenase (ADH) were analyzed in unblanched and blanched 15-mm leek slices packed in atmospheric air (21% O2) or 100% nitrogen (0% O2) three times during 12 months of frozen storage (12 M). The total amount of sulfur compounds and the total amount of aldehydes were greatly influenced by storage time, atmosphere, and blanching [concentration of sulfur compounds in fresh unblanched (UNB) slices = 1.35 mg/L, fresh blanched (B) slices = 1.09 mg/L, UNB 21% O2 12 M = 0.656 mg/L, UNB 0% O2 12 M = 2.11 mg/L, B 21% O2 12 M = 1.14 mg/L, B 0% O2 12 M = 1.59 mg/L]. B 0% O2 was closest to the original ratio between sulfur compounds and aldehydes after 12 months. The activities of HPL and alliinase were totally lost after 12 months, and ADH showed minimal activity, whereas LOX (UNB 0% O2) showed approximately 25% of the original activity. LOX was the most and HPL the least heat labile enzyme investigated.