Recent advances in caffeine and theobromine toxicities: a review

Caffeine and theobromine are purine alkaloids widely consumed as stimulants and snacks in coffee and cocoa based foods and most often as part of ingredients in drugs. Man has enjoyed a long history of consumption of caffeine and theobromine. Recent interest in these two alkoloids, however, is centered on their potential reproductive toxicities. Caffeine and theobromine are now known to cross the placental and blood brain barrier thus potentially inducing fetal malformation by affecting the expression of genes vital in development. The developing fetus may not have developed enzymes for detoxification of these methylxanthine alkaloids via demethylation. There is a need, therefore, to protect the conceptus against insults from teratogens of this nature. Apart from its reproductive toxicity, the presence of caffeine and theobromine in cocoa could limit its potentials as a nourishing food. This is an issue that needs to be addressed by nutritionists and the food industry at large. This paper discusses the natural sources, consumption and uses, toxicity and the major advances in the reproductive toxicology of caffeine and theobromine. The biosynthesis of these compounds in plants, metabolism in mammalian systems and the involvement of cytochrome P450 are reviewed and summarized. Evidence in favor of the toxicity of these compounds in experimental animals is presented with emphasis on the implications of these findings in humans. The paper concludes with a call for caution in the use of caffeine and theobromine pending further and more elaborate investigations.     

茶树咖啡碱合成相关基因研究及利用进展

咖啡碱存在于茶中,由于咖啡碱对一些人引起失眠等一系列副作用,培育低咖啡碱茶树品种具有十分重要的意义。本文综述了近年来咖啡碱生物合成途径中相关酶的基因克隆和功能研究进展,并对其利用现状进行了讨论。     

咖啡因生物合成研究进展及在茶树育种中的应用

咖啡因是一种嘌呤生物碱,存在于茶和咖啡中。由于咖啡因对一些人有引起失眠等的副作用,所以培育低咖啡因咖啡和茶树品种具有非常重要的意义。本综述了近年咖啡因生物合成与代谢的研究进展,包括相关酶的基因克隆等,并对培育脱咖啡因茶树可能的途径和存在问题进行了讨论。     

Microbial and Biochemical Changes Occurring During Production of masvusvu and mangisi,traditional Zimbabwean beverages

Biochemical and microbiological changes that occur during production of masvusvu and mangisi, both traditional Zimbabwean beverages made from malted finger millet flour were investigated in this study. During cooking to produce masvusvu, amylase was active within the first 20 minutes but its activity decreased drastically at temperatures of above 60 °C. Free reducing sugars increased from 3.8 to 308.1 mg/ml. During fermentation to produce mangisis, total aerobic mesophilic bacteria, lactic acid bacteria and yeast and molds increased with fermentation time. Total titratable acidity increased from 0.13 to 0.67 percent, lactic acid from 0.51 t 4.10 g/l and pH decreased from 6.10 to 3.98.     

茶树咖啡碱代谢及低咖啡碱茶树育种研究进展

咖啡碱是茶树中最重要的次生代谢产物之一,一般为茶叶干物质重的2%～5%。咖啡碱具有兴奋神经、祛除疲劳及增加心血管系统活动等保健功能,但摄入过高含量咖啡碱也会引起副作用,因此,研究茶树咖啡碱代谢途径及其分子机理,培育低咖啡碱茶树品种具有非常重要的意义。本文综述了茶树咖啡碱的合成与分解代谢及其关键酶、以及近年来低咖啡碱茶树育种等方面的研究进展,并对这些研究领域目前存在的问题和今后努力的方向进行了讨论和展望。     

茶树咖啡碱合成酶基因mRNA表达的研究

采用原位杂交技术对茶树咖啡碱合成酶基因mRNA的表达进行了研究。结果表明,茶树叶片的表达主要在栅栏组织,该组织的细胞质、细胞核上均有表达,而液泡却没有;叶主脉表达分布在薄壁组织、韧皮部,但韧皮部表达稍弱;幼茎表达分布在韧皮部;不同品种之间的表达位置基本相似、表达信号却有差异。结果还显示,春梢第一片叶比秋梢第一片叶表达信号强;加工技术上绿茶摊青过程0.5 h、1 h、2 h有明显表达信号,而红茶萎凋6 h、8 h、10 h及乌龙茶摇青过程6 h、8 h、10 h的叶子则看不到。     

咖啡碱的生理功能及其作用机制

咖啡碱是世界上消费最广泛的精神类药物及日常饮食成分之一,对机体的神经、内分泌、呼吸和心血管等系统均有复杂的影响,而且咖啡碱还具有抗癌效果。此外,摄入咖啡碱会增加骨质疏松和流产的风险。本文就近年来咖啡碱的生理功能及其作用机制等的相关研究作一简要综述。     

Hydrocyanate,oxalate, phytate,calcium and zinc in selected brands of Nigerian cocoa beverage

Hydrocyanate, oxalate, phytate, calcium and zinc were determined in five brands of cocoa beverage which were coded NC, BT, PN, CT and CA. Hydrocyanate ranged from 5.40 to 9.64 mg/100 g dry matter (DM), oxalate 68 to 146 mg/100 g DM, phytate 590 to 750 mg/100 g DM, calcium 28.7 to 116.4 mg/100 g DM and zinc 0.516 to 0.675 mg/100 g DM. The computed phytate:zinc, calcium:phytate and [calcium] [phytate]/[zinc] molar ratios ranged from 89 to 132, 0.80 to 3.01 and 0.64 to 3.03 respectively. The discussion is focused on toxic levels of hydrocyanate and oxalate, and the significance of the molar ratios in predicting the bioavailability of dietary zinc.     

Effect of soaking,cooking and germination on the oligosaccharide content of selected Nigerian legume seeds

The identity and quantity of and effect ofprocessing on raffinose oligosaccharides in raw,mature seeds of lima beans (Phaseolus lunatus),pigeon peas (Cajanus cajan), African yam beans(Sphenostylis sternocarpa) and jackbeans (Canavalia ensiformis) were investigated. Sucrose,raffinose, stachyose and verbascose were identified byHPLC in all the legume seeds. The total-galactoside contents of the seeds in decreasing order were African yam beans3.84 mg/100 mg; white lima beans 3.62 mg/100 mg; creampigeon peas 3.51 mg/100 mg; red lima beans3.37 mg/100 mg; jackbeans 2.83 mg/100 mg and brownpigeon peas 2.34 mg/100 mg. The predominantoligosaccharide was verbascose in pigeon peas andstachyose in the other three legumes. Cooking unsoakedseeds brought about a greater reduction in the total-galactoside content than soaking for ninehours. The removal of oligosaccharides was higher inlegumes cooked in alkaline solution than in water.Germination quantitatively reduced raffinose,stachyose and verbascose while sucrose was increasedin all seeds except red lima beans and jackbeans.     

Relationship between dietary fiber levels and protein digestibility in selected foods as determined in rats

Samples of 15 food products and feces obtained by feeding them to rats were analysed for dietary fiber fractions. The food products were added as the sole source of protein in 8% protein diets, making up 8.8–51.6% of the diets. Diets were supplemented with 0.54–5.00% purified cellulose to make them more comparable in total fiber. Fiber analyses of food products revealed that the protein sources provided 0.06–7.27% total dietary fiber. The true protein digestibility in rats was negatively correlated with the total food fiber level (r=–0.69,P<0.01) or with the food cellulose level (r=–0.82,P<0.01) but it was positively correlated (r=+0.81,P<0.01) with the purified cellulose level. No relationship was found between protein digestibility and fiber fermentability. Results indicate that several food fiber fractions and possibly associated substances influenced protein digestibility. Purified cellulose did not have the same physiological behavior as food cellulose from the viewpoint of protein digestibility and fiber fermentability.     

贵州绿茶中咖啡碱和儿茶素含量分析

对贵州省243份绿茶样品中咖啡碱和儿茶素类物质采用高效液相色谱方法进行检测。结果表明：贵州省绿茶中儿茶素含量范围为9.14%~27.28%,平均含量15.71%;咖啡碱含量为1.08%~3.33%,平均含量2.24%。各地区茶叶儿茶素品质指数多数为1000~2000,其中黔南地区的最高为1975.51;各地区的儿茶素苦涩味指数主要在8.00~17.04之间,黔西南地区的苦涩味指数最低为8.00。据此分析可得,贵州绿茶具有很高的利用价值,春茶适合开发高品质的名优茶,秋茶适合用于茶叶深加工或有效成分的提取。     

Evaluation of selected food attributes of four advanced lines of ungerminated and germinated Nigerian cowpea (Vigna unguiculata (L.) Walp.)

The chemical composition and functional properties of ungerminated and germinated seeds of four advanced lines of cowpeas (IT81D-699, IT82E-18, IT84S-2246-4 and TVx 3236) were investigated. Ungerminated seeds contained 20.1 to 25.8% crude protein, 2.0 to 2.2% lipid, 115.1 to 210.0 mg phyticacid/100 g and 8.9 to 9.6 mg iron/100 g. The polyphenol contents of the brown- and cream-colored beans were similar (192.0 to 196.0 mg/100 g) but were significantly (p<0.05) higher than the amount (99.8 mg/100 g) found in the white beans. Germination increased the crude protein content, total phosphorus content, nitrogen solubility, fat absorption capacity and foam capacity but decreased ether extract, polyphenol and phytic acid contents, water absorption capacity, bulk density and foam stability of all cowpea lines. Incorporation of up to 0.2 M NaCI improved foam volume. Ungerminated seeds had high water (2.3–3.2 g/g) absorption capacities, while germinated seeds had high fat (3.1–3.6 g/g) absorption capacities. TVx 3236 and IT81D-699 might be selected for combining such characteristics as high crude protein, low phytic acid content and good foaming properties into a single cowpea line through breeding.     

Nitrogen use efficiency in selected rice (Oryza sativa L.) genotypes under different water regimes and nitrogen levels

Water and nutrient availability are two major constraints in most rice-based rainfed shallow lowland systems of Asia. Both stresses interact and contribute to the low productivity and widespread poverty in this environment. The objective of this study was to improve the understanding of interaction between the two factors and to identify varietal characteristics beneficial for productivity in a water- and nutrient-limited rice environment. For this purpose, we screened 19 rice genotypes adapted to different rice environments under two water and two nutrient treatments during the wet season of 2004 and 2005 in southern Luzon, Philippines. Across all genotypes tested and in comparison with the irrigated control, rainfed conditions reduced grain yield of the treatment without N application by 69% in 2004 and by 59% in 2005. The mean nitrogen fertilizer response was highest in the dryer season of 2004 and the rainfed treatment, indicating that water stress had no effect on fertilizer response. Nitrogen application reduced the relative yield loss to 49% of the irrigated treatment in 2004 and to 52% of the irrigated treatment in 2005. Internal efficiency of N (IEN) and recovery efficiency of applied N (REN) were significantly different between genotypes, but were not affected by water availability (REN) or by water and nutrient availability (IEN). In contrast, grain yield and total N uptake were affected by cultivar, N and water availability. Therefore, germplasm for rainfed environments should be screened under conditions of limited and good nitrogen and water supplies. The four best cultivars, CT6510-24-1-2, IR55423-01, IR72, and IR57514-PMI5-B-1-2, performed well across all treatments and both years. Except for IR72, they were all characterized by medium height, medium duration, high early vigor, and a moderate level of drought tolerance. This combination of characteristics seems to enable the optimal use of limited water and nutrient resources occurring in many shallow rainfed lowlands. We also concluded that moderate drought stress does not necessarily affect the response to moderate N rates, provided that drought does not induce high spikelet sterility and that fertilizer N is properly managed.     

玉米单向异交不亲和基因研究进展及其在玉米育种上的应用

玉米单向异交不亲和基因是一种配子体基因(Gametophyte factors,Ga)。Ga基因是影响单倍体配子有性传递的遗传因子,许多带有Ga基因的爆裂玉米可以给不带有Ga基因的马齿型和硬粒型玉米正常授粉结实,反交则通常不结实。Ga基因的这种特性在生产上可以用于特用玉米与普通玉米以及非转基因玉米与转基因玉米之间的生殖隔离,在基础理论上可以用于研究玉米授精过程中的基因互作、信号传导以及雌雄配子间的不亲和机理。     

咖啡碱与儿茶素组合对小鼠体重和脂类代谢的影响

采用不同浓度咖啡碱与儿茶素组合粉末饲料喂养小鼠12周,通过称量小鼠体重、脏器及腹腔脂肪（IPAT）等器官重量,测定血清中的生化指标和肝脏中脂类含量的方法,研究不同浓度咖啡碱与儿茶素组合对小鼠体重、脂类代谢的影响。结果表明,所有实验组的小鼠体重增加与对照组相比有下降趋势,其中Ⅳ组（0.06%咖啡碱+0.6%儿茶素）从第4周开始小鼠体重增加显著减少。咖啡碱与儿茶素组合处理的IPAT重量与对照组相比均明显减少。Ⅰ组（0.03%咖啡碱+0.3%儿茶素）、Ⅱ组（0.03%咖啡碱+0.6%儿茶素）可显著降低血清中游离脂肪酸（NEFA）浓度;Ⅲ组（0.06%咖啡碱+0.3%儿茶素）、Ⅳ组（0.06%咖啡碱+0.6%儿茶素）与对照组相比血清总胆固醇（TC）、甘油三酯（TG）与瘦素浓度明显降低;咖啡碱与儿茶素组合能降低肝脏中TC或TG浓度。以上结果表明,绿茶成分咖啡碱与儿茶素组合通过降低血液与肝脏中的脂类含量,可能引起体内脂肪沉积减少,抑制体重增加。     

普洱茶渥堆发酵中可降解咖啡碱真菌菌株的筛选和鉴定

为了在普洱茶渥堆发酵中筛选出可降解咖啡碱的菌株,采用咖啡碱液体筛选培养基从不同阶段普洱茶发酵样中筛选目标菌株,结合菌落特征、分生孢子结构以及18 S r DNA序列对目标菌株进行鉴定。并将目标菌株接种至茶叶内进行单菌种发酵,通过高效液相色谱法(HPLC)测定咖啡碱、茶碱、可可碱、3-甲基黄嘌呤等嘌呤碱含量。结果表明,在普洱茶发酵样中筛选出一株有效降解咖啡碱的菌株,此菌株在48 h内对咖啡碱的降解率为87.7%。经鉴定为聚多曲霉(Aspergillus sydowii),序列相似性为99.8%。在聚多曲霉茶叶单菌种发酵中,咖啡碱含量急剧下降,在发酵结束时仅占干物质重的(0.414±0.077)%;茶碱质量分数大幅度上升,由最初的不足0.1%上升至(2.129±0.246)%,成为主要的嘌呤碱;可可碱含量无显著变化(P0.05);3-甲基黄嘌呤有大幅度增加,增幅达130%。本文首次从发酵茶叶中筛选出可降解咖啡碱的菌株,探究了微生物发酵中咖啡碱的转化产物,为低咖啡碱普洱茶的开发提供了菌种。     

茶树单宁酶CsTA的理化性质及其在绿茶饮料中的应用

茶叶中的酯型儿茶素对于绿茶滋味有着重要的影响,但是夏秋季节茶树中积累过多的酯型儿茶素会降低茶叶感官品质。前期发现的茶树单宁酶CsTA具有水解酯型儿茶素的功能,但其理化性质以及能否在绿茶饮料加工中发挥一定作用尚不清楚。在前期试验结果的基础上利用液相色谱技术对CsTA进行进一步探究。结果表明,单宁酶CsTA的最适反应温度为45℃、热稳定范围在60℃以下、最适反应pH为7.0,Mg²⁺对单宁酶CsTA活性有明显的激活作用,Cu²⁺可使其完全失活,Mn²⁺、Zn²⁺、Al³⁺、Fe³⁺、Fe²⁺、Li⁺对单宁酶CsTA活性具有一定抑制作用,CsTA在冷冻干燥后有很好的贮存稳定性。将茶树单宁酶CsTA应用于绿茶茶汤后发现,当茶树单宁酶CsTA添加量为11μg、料液比为1∶100、作用时间20 min、温度为35℃时对绿茶茶汤中酯型儿茶素的降解效果最好。本研究为今后利用CsTA在绿茶饮料生产中降低苦涩味,提升品质提供理论参考...     

Nutritional and toxic factors in selected wild edible plants

Nutritional (ascorbic acid, dehydroascorbic acid and carotenes); antinutritional and toxic components (oxalic acid, nitrate and erucic acid) were determined in sixteen popular species of wild edible plants which are collected for human consumption in southeast Spain. Ascorbic+dehydroascorbic acids contents were very high in several species, especially in Chenopodium album L. (155 mg/100 g). Carotenoid content ranged from 4.2 mg/100 g (Stellaria media Villars) to 15.4 mg/100 g (Amaranthus viridis L.). A range of values was found for oxalic acid from absence to 1100 mg/100 g of plant material. Nitrate contents ranged from 47 mg/100 g ( Salicornia europaea L.) to 597 mg/100 g (Amaranthus viridis L.). Low amounts of erucic acid were found in the Cruciferae family (Sisymbrium irio L. 1.73%; Cardaria draba L. 1.23%) and Plantago major L. 3.45%.     

钾肥施用对菜用大豆和普通大豆开花后氮素积累的影响

钾素营养充足,可以提高作物抗逆能力和产量。随着氮、磷肥在生产中施用的增加,钾肥已逐渐成为作物产量的最大限制因子。大豆开花后籽粒形成期是大豆氮素积累、产量提高的重要时期,文章对钾肥施用后菜用大豆和普通大豆两者开花后氮素的积累进行了比较研究。在正常氮磷种肥用量基础上,设置3种施钾处理:不施钾(K0)、种肥施钾120 kg·hm~(-2)(K1)、种肥施钾120 kg·hm~(-2)且在花、荚期喷施30 kg·hm~(-2)叶面钾肥(K2),探究菜用大豆、普通大豆开花后28~56 d内籽粒、叶片、茎中氮素积累动态。结果表明:施用钾肥增进两种类型大豆植株各部位中氮素积累。同时期内,两种类型大豆在各施肥处理下各部位的氮素积累量均为K2K1K0,大豆植株各部位中相对氮素累积量均为籽粒叶片茎。钾肥施用对提高菜用大豆籽粒氮素含量的效应高于普通大豆,与K0相比,K2处理下菜用大豆、普通大豆籽粒平均氮素含量分别增加了0.19%和0.1%。施用钾肥提高了菜用大豆叶片氮素转移效率,相比K0处理,K1、K2处理分别增加了6.1%、8.2%,而对普通大豆影响不大。钾肥施用显著增加普通大豆茎中氮素积累,但菜用大豆品种间差异较大。     

合成茶树咖啡碱相关的N-甲基转移酶基因家族的克隆及序列分析

咖啡碱是茶叶中重要的功能成分,它以黄苷为底物,以S-腺苷甲硫氨酸（SAM）为甲基供体,通过N-甲基转移酶（NMT）类催化的一系列甲基化反应合成的产物。根据NMT基因高度相似的特性,利用长片段PCR法和侧翼序列克隆技术分离了白叶一号6种NMTs的基因组DNA全长,其中有2种为已报道的茶树咖啡碱合成酶基因TCS1、TCS2,1种为假基因,另3种基因分别被命名为TCS3、TCS4、TCS5,基因结构分析发现这6种基因均由4个外显子和3个内含子组成。山茶属植物的NMTs可聚为5类,其中TCS4和TCS5为与其他基因的相似性相对较低的一类。这些结果为今后更好地从基因组水平上剖析茶树咖啡碱的遗传机制提供有用参考。