Role of caffeine and tannin in anti-toxigenic properties of coffee and tea

Aflatoxins B₁, B₂, G₁ and G₂ produced by Aspergillus parasiticus var. globosus IMI 120920 in detannin-caffeinated coffee and black tea were five times more concentrated than in normal tea and coffee. Extracts of normal coffee and tea powders significantly reduced aflatoxins production in liquid broth at 1 and 3 % concentrations, with tea extract. having a more pronounced effect than coffee extract Relevant anti-aflatoxigenic properties appear to be due to tannin and caffeine. These induced 95 % inhibition in aflatoxins at 0.3 % and 0.6 %, respectively. Roasting of contaminated coffee beans at 200 °C for 20 min is effective in the reduction of aflatoxins.     

Biosynthesis and metabolism of purine alkaloids in leaves of cocoa tea (Camellia ptilophylla)

The biosynthesis and metabolism of purine alkaloids in leaves ofCamellia ptilophylla (cocoa tea), a new tea resource in China, have been investigated. The major purine alkaloid was theobromine, with theophylline also being present as a minor component. Caffeine was not accumulated in detectable quantities. Theobromine was synthesized from [8-¹⁴C] adenine and the rate of its biosynthesis in the segments from young and mature leaves from flush shoots was approximately 10 times higher than that from aged leaves from 1-year old shoots. Neither cellfree extracts nor segments fromC. ptilophylla leaves could convert theobromine to caffeine. A large quantity of [2-¹⁴C] xanthine taken up by the leaf segments was degraded to¹⁴CO₂ via the conventional purine catabolic pathway that includes allantoin as an intermediate. However, small amounts of [2-¹⁴C] xanthine were also converted to theobromine. Considerable amounts of [8-¹⁴C] caffeine exogenously supplied to the leaf segments ofC. ptilophylla was changed to theobromine. These results indicate that leaves ofC. ptilophylla exhibit unusual purine alkaloid metabolism as i) they have the capacity to synthesize theobromine from adenine nucleotides, but they lack adequate methyltransferase activity to convert of theobromine to caffeine in detectable quantities, ii) the leaves have a capacity to convert xanthine to theobromine, probably via 3-methylxanthine.     

Degradation and Utilization of Xylans by the Rumen AnaerobePrevotella bryantii(formerlyP. ruminicolasubsp.brevis) B14

Freshly harvested whole cells from cultures ofP. bryantiiB₁4 grown with oat spelt xylan (OSX) as an energy source showed less than 25% of the enzyme activity against OSX, and less than 15% of the activity against birchwood xylan (BWX) and carboxymethylcellulose, that was detectable in sonicated cell preparations. This indicates that much of this hydrolytic activity is either periplasmic, membrane-associated or intracellular and may be concerned with the processing of transported oligosaccharides.P. bryantiiB₁4 cultures were able to utilise up to 45% and 51% of the total pentose present in OSX and BWX, respectively, after 24 h, but could utilize 84% of a water-soluble fraction of BWX. Analysis of the xylan left undegraded after incubation withP. bryantiishowed that while xylose and arabinose were removed to a similar extent, uronic acids were utilized to a greater extent than xylose. Predigestion of xylans with two cloned xylanases from the cellulolytic rumen anaerobeRuminococcus flavefaciensgave little increase in overall pentose utilization suggesting that externalP. bryantiixylanases are as effective as the clonedR. flavefaciensenzymes in releasing products that can be utilised byP. bryantiicells. The xylanase system ofP. bryantiiis able to efficiently utilise not only xylo-oligosaccharides but also larger water-soluble xylan fragments.     

Photoassimilatory and Photorespiratory Behaviour of Certain Drought Tolerant and Susceptible Tea Clones

Net photosynthetic rate (P _N) in the mother leaves was higher in the drought tolerant (DT) clones of tea (Camellia sinensis) while liberation of the fixed ¹⁴C in light from the mother leaves was higher in the drought susceptible (DS) clones. The DT clones translocated more photosynthates to the crop shoots (three leaves and a bud) from the mother leaf than the DS clones. Concentrations of RuBP carboxylase (RuBPC) or oxygenase (RuBPO) had no relationship with the drought tolerant nature of tea clones but their ratio correlated with the same. DT tea clones had higher catalase activity that could scavenge the hydrogen peroxide formed in the photorespiratory pathway and thereby reduced photorespiration rate (P _R). The ratio of RuBPC/RuBPO had a positive correlation with P _N and catalase activity. Negative correlation between RuBPC/RuBPO and P _R and between catalase activity and RuBPO activity was established.     

茶树CsPLK基因的鉴定与表达分析

茶树中富含茶氨酸、儿茶素和咖啡碱等重要功能成分,具有较高的价值功效,茶树在生命周期中经常遭受逆境胁迫,维生素B6(VB6)在植物体内参与逆境应答,吡哆醛激酶(pyridoxal kinase,PLK)是VB6补救途径中的关键酶。为进一步了解PLK在茶树生物合成中的功能和作用机理,该研究基于茶树基因组数据库,以龙井43为材料,采用逆转录PCR(RT-PCR)的方法从茶树中克隆出CsPLK的基因。结果表明:该基因序列长为1 179 bp,编码393个氨基酸; CsPLK蛋白和已知物种中PLK蛋白具有较高的同源性,都是核糖激酶超家族成员;通过构建pET-CsPLK载体进行原核表达,并鉴定出重组蛋白有很强的催化活性;组织表达特异性分析表明,叶中的表达量比茎、根的高,在根中最低;荧光定量PCR表示,低温诱导CsPLK上调表达,干旱诱导CsPLK下调表达,发现该基因在茶树中有明显的逆境应答,推测CsPLK在茶树的生长发育、逆境胁迫发挥重要作用。     

Evaluation of antioxidant properties of medical plants using microbial test systems

The antioxidant activities of extracts from leaves of the medicinal plants growing in Siberia were examined. Total antioxidant activity was determined using in vitro methods including DPPH (2,2-diphenyl-1-picrylhydrazyl radical) free radical scavenging assay, chelating capacity assay with ferrozine, evaluation of capacity to protect plasmid DNA against oxidative damage, measurement of H₂O₂ production, and measurement of total flavonoid and tannin content as well. Using in vivo experiments, we also evaluated capacities of the plant extracts to protect bacteria Escherichia coli against bacteriostatic and bactericidal effects of H₂O₂, and influence of the plant extracts on expression of antioxidant gene katG, encoding catalase. The extracts from Chamerion angustifolium, Filipendula vulgaris and Pyrola rotundifolia indicated the highest levels of antioxidant activity both in vivo and in vitro. Our data suggest that the extracts of the tested plants may provide antioxidant effects on bacteria simultaneously through different pathways, including direct radical scavenging, iron chelation and induction of genes encoding antioxidant enzymes.     

氮素水平对茶树叶片氮代谢关键酶活性及非结构性碳水化合物的影响

以福鼎大白茶(FD)、保靖黄金茶1号(HJ1)、白毫早(BHZ)为材料,设置不施氮N_0(0 g)、低氮N1(11 g)、中氮N_2(22 g)和高氮N_3(33 g)4个氮素水平的盆栽实验,研究了铵态氮对3个品种茶树的根系活力、氮代谢关键酶及非结构性碳水化合物(NSC)的影响。结果表明:随着施氮水平的提高,N_2、N_3处理的茶树根系活力较对照N0显著增加(P0.05),但二者间无显著差异;叶片谷氨酰胺合成酶(GS)、谷氨酸合成酶(GOGAT)活性总体呈上升趋势;与对照比较,茶树叶片全氮和可溶性蛋白含量增加,其中HJ1在N_2和N_3处理后显著增加(P0.05);在3个茶树品种中,非结构性碳水化合物中可溶性总糖含量均呈上升趋势,淀粉含量具有品种特异性,施氮处理后3个茶树品种氮代谢关键酶活性及NSC含量变化存在差异,以HJ1的氮同化关键酶GS、GOGAT酶活性较高、根系活力较强,氮代谢产物显著增加,表明其具有较高的氮同化速率。施氮后HJ1的总NSC的含量及碳氮比的变化幅度较另外2个品种小,能够更好的保持碳氮平衡,游离氨基酸含量增幅较高,品质更优。因此,通过茶树氮代谢关键酶活性及非结构性化合物的研究能为茶树品种的品质评价以及提高茶树的品质和氮素利用效率提供依据。     

Effects of processing methods on nutritional composition and antioxidant activity of mealworm (Tenebrio molitor) larvae

We examined the effects of different processing methods on the nutritional composition and antioxidant activity of mealworms. After processing with nine methods, we calculated the contents of protein, fat, ash, carbohydrate, minerals (P, Ca, K, Fe, Na), vitamin B group (B₁, B₂, B₃), moisture, and calories. When processed by freeze drying among freeze drying, hot air drying, oven broiling, roasting, pan frying, deep frying, boiling, steaming, and microwaving, the contents of protein, some minerals, and vitamins were the highest. The content of total minerals was lowest after deep frying, and those of vitamin B₁ and B₃ were the lowest after microwaving. Antioxidant activity was then evaluated using DPPH and ABTS radical scavenging assays. DPPH assays showed that microwaving, freeze drying, deep frying, steaming, boiling, and oven broiling of mealworms yielded scavenging activities of 20.9–29.0% at 2,000 μg/mL, which was similar to the activity level (22.7–33.2%) of 40–60 μM tocopherol. ABTS assays confirmed that only freeze‐dried mealworms at 2,000 μg/mL exhibited higher activity than 10 μM tocopherol. Interestingly, similar trends were found for antioxidant activity levels and total phenolic contents in mealworms.     

<Emphasis Type="Italic">Ilex Vomitoria</Emphasis> Ait. (Yaupon): A Native North American Source of a Caffeinated and Antioxidant-Rich Tea

Ilex Vomitoria Ait. (Yaupon): A Native North American Source of a Caffeinated and Antioxidant-Rich Tea. Yaupon holly (Ilex vomitoria Ait.) is a caffeine-containing shrub native to the southeastern United States where its leaves and twigs were traditionally used to prepare a stimulating and healthful beverage by Amerindians and more recent colonists. For a variety of mostly socioeconomic and cultural reasons, widespread consumption of yaupon tea ceased by the late 19th century, but the species is widely used in ornamental horticulture. Given the environmental damage associated with other caffeine crops, we believe that disuse of this species is unfortunate, and we report on traits that consumers may consider valuable. We found that total foliar biomass, caffeine, and antioxidant production all increased with nitrogen fertilization in one common ornamental yaupon cultivar, ‘Nana.’ Increasing light availability was associated with increased antioxidant activity but not with the decreased caffeine production predicted by the carbon/nutrient balance hypothesis for secondary metabolite production. We also found the highest caffeine concentrations in another yaupon cultivar, ‘Pendula,’ but suggest that the wide range of chemical variation offered by wild-type yaupon populations renders them more suitable as sources for the development of high caffeine-producing varieties. The results of this study suggest that yaupon is a viable caffeine alternative for North Americans living within its range on the southeastern coastal plain.     

Effects of a Component of Green Tea on the Proliferation of Vascular Smooth Muscle Cells

The effects of a component of green tea on the proliferation of smooth muscle cells were measured in terms of [³H]thymidine uptake. When green tea tannin mixture was added to the medium of cultured smooth muscle cells, it suppressed the proliferation of the cells dose-dependently. Similarly to the effects of the green tea tannin mixture, (–)-epigallocatechin 3-O-gallate, its main ingredient, had an inhibitory effect on smooth muscle cell proliferation at a low concentration. (–)-Epicatechin 3-O-gallate was also an effective component. Among four types of gallate-free tannin, (–)-epigallocatechin, (–)-epicatechin, and (+)-catechin showed significant dose-dependent inhibition of smooth muscle cell proliferation. However, caffeine and theanine were found to have no such action.     

Green tea (Camellia sinesis) ameliorates female Schistosoma mansoni-induced changes in the liver of Balb/C mice

This study was designed to assess the effect of green tea, an aqueous extract of Camellia sinensis, on the oxidative stress, antioxidant defense system and liver pathology of Schistosoma mansoni-infected mice. Green tea at concentration of 3% (w/v) was given orally to treated mice as sole source of drinking water from the end of the 4th week to the end of 10th week post-infection; untreated mice were allowed to drink normal water. The data of the studied S. mansoni-infected mice exhibited a suppression of hepatic total antioxidant capacity, superoxide dismutase (SOD), catalase (CAT) activity and glutathione content. The liver lipid peroxidation was deleteriously elevated in S. mansoni-infected mice. The hepatic total protein content, AST and ALT activities were profoundly decreased in the S. mansoni-infected mice. Most hepatocytes were damaged and showed abnormal microscopic appearance with aggressive necrosis. Both total protein and glycogen levels have been greatly reduced as indicated by histochemical examination. The treatment of S. mansoni-infected mice with green tea succeeded to suppress oxidative stress by decreasing the lipid peroxides but failed to significantly enhance the antioxidant defense system and deteriorated changes owing to liver damage and necrosis. In consistence with biochemical data, histopathological and histochemical data indicated that treatment of S. mansoni-infected mice with green tea could ameliorate hepatocytes thus reduce cellular necrosis and partially restore both total protein and glycogen levels. Thus, the study concluded that the green tea suppresses the oxidative stress through its constituent with free radicals scavenging properties rather than through the endogenous antioxidant defense system.     

沙芥属植物活性氧清除系统对干旱胁迫的响应

以沙芥属植物沙芥和斧形沙芥幼苗为试材,采用盆栽控水干旱方法,分析其在不同干旱胁迫强度下根和叶的活性氧水平、抗氧化酶活性和抗氧化剂含量的变化,并利用隶属函数法和抗旱系数法综合评价沙芥和斧形沙芥的抗旱性。结果表明: （1）随着干旱胁迫程度的加剧,沙芥和斧形沙芥的根和叶中O^-·₂产生速率及·OH、H₂O₂、MDA含量总体呈逐渐升高的趋势,且干旱胁迫下沙芥比斧形沙芥产生了更多的ROS和MDA。（2）随着干旱胁迫程度的加剧,沙芥和斧形沙芥的根和叶中POD、APX、GST活性及叶中GR活性均先升高后降低,叶中的SOD活性以及根中GR、GPX活性均先降低后升高,根和叶中的CAT活性、叶中的GPX活性和根中SOD活性均逐渐升高;但根和叶中的SOD、POD、CAT活性在各干旱处理下均表现为斧形沙芥高于沙芥。（3）沙芥和斧形沙芥的根和叶中AsA含量随着干旱胁迫程度的加剧而先升高后降低,GSH含量逐渐升高,CAR含量逐渐降低,而VE含量在叶中逐渐升高,在根中却逐渐降低;但斧形沙芥比沙芥合成更多的AsA和GSH,其植物体内AsA GSH循环系统能清除更多的ROS。（4）沙芥和斧形沙芥的根和叶中总抗氧化能力（T AOC）均随着干旱加剧逐渐增强,且斧形沙芥的总抗氧化能力强于沙芥;活性氧清除系统的平均隶属度和综合抗旱系数显示,轻度干旱胁迫下沙芥抗旱性强于斧形沙芥,中度和重度干旱胁迫下斧形沙芥的抗旱性强于沙芥。研究认为,在干旱胁迫条件下,斧形沙芥根叶中ROS和MDA含量明显低于沙芥,而其大部分抗氧化酶活性和抗氧化剂含量高于沙芥,斧形沙芥植株体内抗氧化系统表现出更强的活性氧清除能力,从而表现出更强的抗旱性。     

Vitamins as effectors of monensin production byStreptomyces cinnamonensis

Vitamins added to submergedStreptomyces cinnamonensis cultures stimulated the production of monensins. Vitamins B₂, B₃, B₅ and B₁₂ enhanced the production by about 50%, vitamins B₁ and B₆ by 100%. The addition of biotin in optimal concentration resulted in more than 3-fold increase in total production.     

Antioxidant Effects of Rhodoxanthin from Potamogeton crispus L. on H2O2-Induced RAW264.7 Macrophages Cells

Potamogeton crispus L. (P. crispus) is the type of a widely distributed perennial herbs, which is rich in rhodoxanthin. In this research work, five antioxidant indexes in vitro were selected to study the antioxidant activity of rhodoxanthin from P. crispus (RPC). A model of hydrogen peroxide (H₂O₂) -induced oxidative damage in RAW264.7 cells was established to analyze the antioxidant effect and potential mechanism of RPC. The levels of ROS, MDA and the activities of oxidation related enzymes by H₂O₂ were determined by enzyme linked immunosorbent assay (ELISA). The mRNA expression of Nrf-2, HO-1, SOD1 and SOD2 was measured by qRT-PCR assay. According to the results, RPC had free radical scavenging ability for 2, 2-diphenyl-1-trinitrohydrazine (DPPH), 2,2’-azinobis(3-ethylbenzo-thiazoline-6-sulfonic acid radical ion) (ABTS), hydroxyl radical and superoxide anion. RPC significantly decreased the level of MDA and ROS and LDH activity, while increased GSH level and activities of SOD, GSH−Px and CAT. It was showed that RPC could increase the mRNA expression of Nrf-2, HO-1, SOD1 and SOD2 in RAW264.7 cells in a dose-dependently manner. In summary, RPC treatment could effectively attenuate the H₂O₂-induced cell damage rate, and the mechanism is related to the reduction of H₂O₂ induced oxidative stress and the activation of Nrf-2 pathway.     

从黑茶中分离和筛选产单宁酶菌株

茶叶中富含单宁化合物。从分离自黑茶的真菌菌株中,筛选高产单宁酶的菌株;进而分离纯化单宁酶,分析单宁酶对茶汤的转溶效果。从不同产地的3个黑茶样品中,共分离获得44个真菌分离物;经初步鉴定,这些真菌分离物以曲霉属（Aspergillus）、青霉属（Penicillium）和散囊菌属（Eurotium）的真菌居多。以单宁酸为底物的鉴别培养基初筛表明,其中26个真菌分离物在鉴别平板上产生透明圈,显示单宁水解酶活性;通过固体发酵复筛,筛选到1株产单宁酶活性较高的菌株,初步鉴定为青霉属（Penicillium）菌株,命名为青霉MP-24菌株。青霉MP-24可以以茶叶、茶梗和麸皮等农副产品作为原料固体发酵产生单宁酶。以麸皮为原料的发酵产物经过硫酸铵分级沉淀、DEAE阴离子交换层析和葡聚糖G-150凝胶层析等分离纯化步骤,得到分子量为70 kDa的单一蛋白质条带,单宁酶活力达到603.68 U/mg。纯化获得的单宁酶对茶汤有良好的转溶效果。研究结果表明,在黑茶相关微生物中含有丰富的产单宁酶菌株,是工业酶制剂的重要资源。     

Effects of Kaolin (M-99-099) Application on Antioxidant and Phenolic Compounds in Tea Leaves (<Emphasis Type="Italic">Camellia sinensis</Emphasis> L.O. Kuntze)

In this study, the effects of kaolin (M-99-099) applications on the total phenolic and antioxidant contents of tea leaves (Camellia sinensis (L.) O. Kuntze) harvested in three different periods were investigated. For this purpose, four different strategies including standard fertilizer application (T), 3% kaolin+standard fertilizer application (Ka₁), 6% kaolin+standard fertilizer application (Ka₂), and control (C) (nothing applied) were used to compare the effects of these strategies on total phenolic content, FRAP values, and DPPH radical scavenging capacities of tea leaves. It has been proven that the kaolin applications, Ka₁ and Ka₂, increase the phenolic content and antioxidant contents of tea samples. While the kaolin applications have higher values at 1st harvest than those of T and C, they have the lowest values at 3rd harvest.     

雪松不同部位中原花青素的含量测定及其清除DPPH自由基的活性

雪松(Cedrus deodara)是松科(Pianaceae)雪松属(Cedrus)树种的泛称,为广泛分布于喜马拉雅山脉的常绿树种,包括雪松(C.deodara)、黎巴嫩雪松(C.libani)、短叶雪松(C.brevifolia)和北非雪松(C.atlantica)等。雪松的针叶药用历史悠久,主要化学成分为黄酮类、苯丙素类、有机酸类、三萜类、甾体类、多糖及针叶胶等,具有抗肿瘤、抗氧化、改善记忆、抗菌及抗病毒等多种功效。为了测定雪松不同部位中原花青素的含量,并考察其清除DPPH自由基活性的能力,该研究以Fe3+离子为催化剂,用正丁醇-盐酸法分别测定雪松松针、花序、小枝、木质部、树皮中原花青素的含量,并对不同部位提取液清除DPPH自由基的能力进行测定。结果表明:松针、花序、木质部、树皮、小枝中原花青素的含量分别为12.93%、12.93%、6.41%、12.00%、3.37%,在质量浓度(以生药计)为0.8 mg·m L-1时,其对DPPH自由基的清除率均达到或接近90%。该研究结果提示雪松各个部位的原花青素含量丰富,其提取液的体外抗氧化活性很高,这为雪松的进一步开发利用提供了很好的参考价值。     

Chemical Characterization and Biological Activities of Phenolic‐Rich Fraction from Cauline Leaves of Isatis tinctoria L. (Brassicaceae) Growing in Sicily,Italy

The present work focused on the evaluation of the antioxidant and cytotoxic activities of the phenolic‐rich fraction (ItJ‐EAF) obtained from cauline leaves collected in January from Isatis tinctoria L. (Brassicaceae) growing wild around Acireale (Sicily, Italy). The total phenolic, flavonoid, and condensed tannin contents of the fraction were determined spectrophotometrically, whereas the phenolic profile was assessed by HPLC‐PDA/ESI‐MS analysis. A total of 20 compounds were positively identified and twelve out of them were never previously reported in I. tinctoria leaves. The fraction exhibited good radical scavenging activity in DPPH test (IC₅₀ = 0.6657 ± 0.0024 mg/ml) and reducing power (3.87 ± 0.71 ASE/ml), whereas, it neither showed chelating activity nor was able to counteract H₂O₂ induced oxidative stress damage in Escherichia coli. The antiproliferative effect was evaluated in vitro on two human anaplastic thyroid carcinoma cell lines (CAL‐62 and 8505C) by MTT assay. At the highest tested concentration ItJ‐EAF significantly reduced (80%) the growth of CAL‐62 cells. No cytotoxicity against Artemia salina was observed. It can be concluded that I. tinctoria cauline leaves represent a source of phenolic compounds which could be potentially used as chemopreventive or adjuvant agents against cancer.     

基于HPLC-ECD研究牡荆叶抗氧化的谱-效关系

为研究牡荆叶指纹图谱与抗氧化活性的谱-效关系，该研究首先建立了18批牡荆叶的高效液相色谱-电化学检测法(HPLC-ECD)指纹图谱，对不同来源牡荆叶药材进行聚类分析，鉴定主要酚类化合物且测定其含量，分析牡荆叶的总酚和总黄酮含量，并采用DPPH自由基清除法、ABTS自由基清除法、氧自由基吸收能力法及铁离子还原能力法考察其体外抗氧化活性，通过皮尔逊相关分析、灰度关联分析及偏最小二乘回归分析法研究牡荆叶的谱-效关系。结果表明：(1)牡荆叶的指纹图谱标定21个共有峰，共指认出10个峰，其含量顺序为绿原酸>异荭草苷>木犀草苷>异牡荆素>异绿原酸A>异绿原酸C>原儿茶酸>荭草苷>异绿原酸B>新绿原酸；不同产地样品间相似性较高，相似度结果为0.816～0.983。(2)系统聚类分析显示，样品含量对分类有一定影响，不同来源样品被分为3类，其中南北方样品存在一定差异。(3)牡荆叶中总酚和总黄酮的含量分别为15.82～61.83 mg·g^-1和27.85～157.65 mg·g^-1,样品均具不同程度的抗氧化活...