免疫亲和柱净化—荧光快速检测技术研究

报道了一种免疫亲和柱净化-荧光法快速检测花生及其制品中黄曲霉毒素B1的分析方法。用70%甲醇对样品进行初步提取，过滤后用石油醚萃取脱脂，萃取液稀释后用免疫亲和微柱纯化富集，洗脱液放入黄曲霉毒素专用荧光定量速测仪器中自动读取结果。该方法直接读数的线性范围为0.3~25µg/kg，R2=0.9995，方法的平均回收率超过90%，变异系数低于5%，最低定量浓度为0.3µg/kg。应用于花生及其制品的黄曲霉毒素B1检测，单个样品检测全过程（含样品提取）所需时间可以控制在45min左右，检测成本低于其他仪器方法。     

胶体金免疫层析法快速定量分析粮油农产品中黄曲霉毒素B1

采用自主研制的黄曲霉毒素B1胶体金免疫定量检测卡,建立花生、玉米、大米、小麦等粮油农产品中黄曲霉毒素B1的定量分析方法,4种样品检测的线性范围为1.0～20.0μg/kg,R2＞0.97,方法的定量限为1.0 μg/kg,样品加标回收率为75%～106%,RSD＜20%。胶体金免疫层析法与免疫亲和柱净化-HPLC法相比,相对误差＜15%,具有简便快速、灵敏度高、重现性好等特点,适用于粮油农产品及制品中黄曲霉毒素B1 筛查,样品检测时间只需15min,检测成本低于其他方法。     

高效液相色谱柱后光化学反应-荧光检测茶叶中黄曲霉毒素B1

建立了高效液相色谱/光化学反应器/荧光检测器测定茶叶中黄曲霉毒素B1的方法。用乙腈水溶液(V∶V=86∶14)提取黄曲霉毒素B1,提取液经净化柱和黄曲霉毒素B1免疫亲和柱净化,高效液相色谱测定。在黄曲霉毒素B1标准溶液质量浓度为0.591~5.91μg/L时,峰面积与浓度呈现良好的线性关系,黄曲霉毒素B1的回收率为85.4%~98.9%(添加量分别为0.510μg/kg、7.090μg/kg和14.180μg/kg),相对标准偏差为0.2%~1.8%,方法检出限为0.1μg/kg。运用所建立方法对市售的8个茶样及加标样品中的黄曲霉毒素B1进行检测,结果显示该方法选择性强、灵敏度高,适合茶叶中黄曲霉毒素B1的测定。     

高效液相色谱柱后光化学反应-荧光检测茶叶中黄曲霉毒素B1

建立了高效液相色谱/光化学反应器/荧光检测器测定茶叶中黄曲霉毒素B1的方法。用乙腈水溶液(V∶V=86∶14)提取黄曲霉毒素B1,提取液经净化柱和黄曲霉毒素B1免疫亲和柱净化,高效液相色谱测定。在黄曲霉毒素B1标准溶液质量浓度为0.591~5.91μg/L时,峰面积与浓度呈现良好的线性关系,黄曲霉毒素B1的回收率为85.4%~98.9%(添加量分别为0.510μg/kg、7.090μg/kg和14.180μg/kg),相对标准偏差为0.2%~1.8%,方法检出限为0.1μg/kg。运用所建立方法对市售的8个茶样及加标样品中的黄曲霉毒素B1进行检测,结果显示该方法选择性强、灵敏度高,适合茶叶中黄曲霉毒素B1的测定。     

环糊精对花生黄曲霉毒素B_1荧光增强作用与应用研究

本文探讨了一种新型无毒的黄曲霉毒素荧光增强剂—β-环糊精及其衍生物。结果表明,在2mL黄曲霉毒素B1溶液中加入1mL0.01mol/Lβ-环糊精或其衍生物时荧光增强倍数最大,其中β-环糊精可使荧光信号增强5倍,2,6-二甲基-β-环糊精荧光增强倍数可达到8倍,表明β-环糊精及其衍生物对黄曲霉毒素荧光增强作用极为显著;利用该增强剂在黄曲霉毒素荧光增强前和荧光增强后的信号变化量和黄曲霉毒素浓度之间的线性关系建立了新型荧光增强剂免疫亲和柱荧光光度检测方法,最低检出限可达0.3μg/kg,相关系数都达到0.99以上;对花生样品的添加回收率在90%～120%之间;与基于高效液相色谱国家检测标准方法比对分析,两种检测方法无显著差异,相对误差小于4%,为黄曲霉毒素高灵敏快速检测技术开辟了新的途径。     

免疫亲和净化-高效液相质谱法检测茯砖茶中黄曲霉毒素B1

为建立检测茯砖茶中黄曲霉毒素B1的免疫亲和净化-高效液相色谱质谱分析方法,并对20份茯砖茶样品中黄曲霉毒素B1含量进行测试.样品经过70％甲醇溶液匀浆提取,免疫亲和柱富集净化,经C18色谱柱分离后采用三重四级杆质谱检测.结果表明,HPLC-MS检测线性关系良好,方法检出限为0.03μg/kg,加标浓度2～20μg/kg...     

中国西南花生产区黄曲霉菌分布、产毒力及花生黄曲霉毒素污染

为探明中国西南花生产区黄曲霉菌分布、产毒力及产后花生黄曲霉毒素污染情况,从云南广南、西藏察隅、四川蓬安采集177份花生、土壤样品,共分离鉴定出黄曲霉菌206株,分析其分布及产毒特征,并调查71份产后花生样品黄曲霉毒素污染情况。结果表明,四川蓬安花生产地黄曲霉菌分布最广,数量最多,其检出率和菌落数分别为50.0%、212.0 CFU/g,西藏察隅黄曲霉菌分布最少（18.1%,52.8 CFU/g）。菌株产毒力研究结果表明,88.6%的菌 株能产生黄曲霉毒素,产毒类型以B族毒素为主,尤其是AFB1,其含量在0～8500μg/L,平均产毒能力排序为：云南广南（4393.4μg/L）>西藏察隅（1991.0μg/L）>四川蓬（1259.3μg/L）;不产毒菌株占11.4%,均来自西藏察隅。对产后花生黄曲霉毒素污染研究发现,西南地区花生黄曲霉毒素污染较轻,阳性样品检出率为7.0%,AFB1含量在0~7.2 μg/kg;四川蓬安花生样品AFB1检出率为4.2%,含量0~7.2 μg/kg,云南广南AFB1检出率为16.7%,含量0~2.1 μg/kg, 西藏察隅样品未检出AFB1。西南产区黄曲霉菌检出率越高、菌落数越多,产后花生黄曲霉毒素污染越严重。     

氯化钙对花生中黄曲霉毒素B1提取及其快速检测结果的影响

本文以氯化钙为蛋白沉淀剂、黄曲霉毒素B1为靶标,探究了70%甲醇水（V/V）提取液中不同含量氯化钙对花生中黄曲霉毒素提取及其免疫检测结果的影响;采用样品加标回收的方法,研究比较了五种不同含量氯化钙对胶体金免疫层析试纸条和ELISA检测结果的影响。研究结果表明,样品经含1% CaCl2 （m/V）的甲醇水处理后,用胶体金免疫层析试纸条检测时,试纸条T线颜色梯度变化,不同浓度加标回收率等参数均得到明显改善,前处理效果显著;选取20份花生实际样品,经过这种前处理之后,进行胶体金免疫层析试纸条和ELISA检测,同时将其检测结果与免疫亲和层析高效液相色谱法（IAC-HPLC）进行比对,胶体金免疫层析试纸条与IAC-HPLC检测结果符合率为90%,间接竞争ELISA与IAC-HPLC检测结果相关性系数达0.996,表明本研究所改进的样品前处理方法能提高免疫检测准确度,具有很好的应用性。     

量子点标记荧光免疫法检测花生中黄曲霉毒素B1

建立了基于量子点标记二抗的间接竞争荧光免疫吸附测定方法（indirect competitive fluorescence-linked immunosorbent assay,cFLISA）,并研究检测花生中黄曲霉毒素B1可行性。采用谷胱甘肽为稳定剂,在水相中直接合成碲化镉（CdTe）量子点,并利用EDC与兔抗鼠二抗进行共价偶联,以黄曲霉毒素B1的单克隆抗体建立cFLISA方法。结果表明,该方法的灵敏度和最低检测限值分别为0.023ng/mL和0.001ng/mL,与传统的有机染料FITC-二抗法比较,灵敏度提高了30倍,花生样品加标0.1、0.05和0.025ng/g,回收率范围在88%~116%之间,变异系数均小于10%。建立的cFLISA方法可以较好的检测花生中黄曲霉毒素B1,并为其它真菌毒素的检测提供参考。     

花生黄曲霉毒素污染臭氧脱毒技术研究

研制了一种花生黄曲霉毒素污染臭氧脱毒专用装置,分析花生黄曲霉毒素污染的臭氧脱毒技术和脱毒效果,包括通气方式、臭氧浓度、处理时间和花生水分含量对脱毒的影响等。结果表明,T-1（即下部通入臭氧气体,上部排气）方式下,臭氧浓度6.0mg/L、处理时间30min、花生水分含量为5%时脱毒效果最佳。在此条件下,花生中黄曲霉毒素总量和B1脱毒百分率分别为65.88%和65.90%,脱毒效果显著。花生黄曲霉毒素污染臭氧脱毒装置和技术操作简单、重现性好、环境友好,适用于花生及粮油制品黄曲霉毒素脱毒。       

In vitro reproduction in the annual pasture legumes subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.)

Pasture legumes are important components of both mixed farming rotations and permanent pastures in temperate climates. Breeding of two widely sown pasture legumes, subterranean clover (Trifolium subterraneum L.) and French serradella (Ornithopus sativus Brot.), is constrained by the long generation cycle, typically enabling only one generation per year. We hypothesized manipulation of culture medium and conditions would enable the development of a laboratory‐based protocol for in vitro reproduction in subterranean clover and French serradella. In vitro flowering and viable seed set was induced from both species. For subterranean clover, the most effective treatment was culturing on modified MS medium with 1 μm kinetin and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and continuous photoperiod. For French serradella, culture on a hormone‐free B5 medium with 5 mm NH₄Cl and 0·1 m sucrose under a 100 μmol m^?2 s^?1 light intensity and 20 h photoperiod was optimum. It is expected this technique will have application in accelerating generation turnover within breeding programs, for the study of factors influencing flowering in pasture legumes, and for the propagation of valuable yet enfeebled plants such as embryo‐rescued hybrids.     

Pharmacokinetics of Marine-Derived Drugs

Marine organisms represent an excellent source of innovative compounds that have the potential for the development of new drugs. The pharmacokinetics of marine drugs has attracted increasing interest in recent decades due to its effective and potential contribution to the selection of rational dosage recommendations and the optimal use of the therapeutic arsenal. In general, pharmacokinetics studies how drugs change after administration via the processes of absorption, distribution, metabolism, and excretion (ADME). This review provides a summary of the pharmacokinetics studies of marine-derived active compounds, with a particular focus on their ADME. The pharmacokinetics of compounds derived from algae, crustaceans, sea cucumber, fungus, sea urchins, sponges, mollusks, tunicate, and bryozoan is discussed, and the pharmacokinetics data in human experiments are analyzed. In-depth characterization using pharmacokinetics is useful for obtaining information for understanding the molecular basis of pharmacological activity, for correct doses and treatment schemes selection, and for more effective drug application. Thus, an increase in pharmacokinetic research on marine-derived compounds is expected in the near future.     

Estimation of lucerne yield stability for enabling effective cultivar selection under rainfed conditions

Evaluation of crop and forage yield stability is of increasing relevance in the context of current and recent environmental changes but, in contrast to other field crops, there are no published systematic analyses among forage crops in Europe. A study of stability performance was conducted with 13 Czech cultivars of lucerne at four locations over a 2‐year period with the following aims: (a) to evaluate yield stability of varieties across different environments and (b) to calculate measurable benefits of variety selection in relation to the specific environment. The cultivar Vlasta was identified as the highest yielding cultivar (annual yield 16.0 t DM/ha), whereas the lowest yielding cv. Magda, Tereza and Oslava averaged around 14.9 t DM/ha. Effect of genotype × environment interaction (G × E) was two times higher than for genotype alone. Additive main effects and the multiplicative interaction (AMMI) model showed that the highest yielding cultivars may not be stable across environments. This study demonstrated further that significant yield improvement could be detected, even among a relatively homogenous group of domestic cultivars, and this was driven mainly by site productivity: the improvement was +10% in low‐yielding sites, compared with +3% in high‐yielding sites. Results highlight that advanced agronomy should also consider stability parameters such as AMMI stability value or superiority measures for forage crops in response to the challenges associated with climate change.     

La pourriture fongique des tubercules de pomme de terre en atmosphère contrôlée

Résumé Fusarium culmorum et, dans une moindre mesure,F. avenaceum, F. sambucinum, Colletotrichum coccodes etCylindrocarpon sp. ont été isolés de tubercules entreposés en atmosphère contr?lée (AC) et atteints de pourriture grave. Des expériences d'inoculation de tubercules avec 10 espèces ou variétés fongiques et conservation en AC à 8 et 10°C ont montré que: 1) les champignons non pathogènes en atmosphère normale (ex.:Cylindrocarpon sp.) le devenaient en AC, 2) ceux qui étaient faiblement pathogènes en atmosphère normale étaient nettement plus virulents en AC (ex.:F. culmorum), 3) les espèces ou variétés virulentes en conditions d'air normales (Fusarium sulphureum etPhoma exigua var.foveata) ne provoquaient pas de pourriture plus grave en AC et leur activité pouvait même être légérement freinée.

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Summary The extensive rotting of tubers which occurred during storage trials in controlled atmospheres has led to a mycological and pathological study of the rots. The most frequently isolated species wasFusarium culmorum althoughF. avenaceum, F. Sambucinum, Coletotrichum coccodes and aCylindrocarpon sp. were also present. Tubers of cvs Bintje and Ulla were inoculated with 19 isolates of 10 fungal species or varieties associated with rotting of stored tubers (Table 1). The tubers were incubated at 8 or 10°C in normal or controlled (2% O₂:1% CO₂, 2% O₂:6% CO₂, 2% O₂:12% CO₂, 6% O₂:6% CO₂) atmospheres and the extent of rotting was determined a month later (Tables 2 and 3 and Fig. 2). Species which were weakly virulent in a normal atmosphere (variousFusarium spp. andPhoma exigua var.exigua) were clearly more aggressive in the controlled atmosphere and this was especially evident in the case ofF. culmorum. Species ot pathogenic in a normal atmosphere under experimental conditions (Colletotrichum coccodes, Cylindrocarpon sp.,Phoma eupyrena) caused rotting in the controlled atmosphere. The two species which were virulent in a normal atmosphere (F. sulphureum andP. exigua var.foveata) were also virulent in the controlled atmosphere and in some cases the extent of rotting was even less. As a general rule, rotting caused by weakly virulent fungi was greater in both cultivars in the controlled than in a normal atmosphere.

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Zusammenfassung Eine starke F?ulnis von in Kontrotrollierter Atmosph?re gelagerten Kartoffelknollen veranlasste eine mykologische und pathologische Untersuchung. Der am h?ufigsten isolierte Pilz warFusarium culmorum. F. avenaceum, F. sambucinum, Colletotrichum coccodes undCylindrocarpon sp. wurden ebenfalls bestimmt. Neunzehn St?mme von 10 Pilzsorten oder-Arten welche bei Lagerf?ulnis vorkommen (Tab. 1) wurden auf Knollen der Sorten Bintje und Ulla inokuliert. Die Lagerung wurde bei 8 oder 10°C under Normalbedingungen oder kontrollierter Atmosph?re durchgeführt (2% O₂:1% CO₂, 2% O₂:6% CO₂, 2% O₂:12% CO₂, 6% O₂:6% CO₂). Der F?ulnisgrad (Abb. 1) wurde nach einem Monat bonitiert. Die Ergebnisse sind in den Tabellen 2 und 3 dargestellt und in der Abb. 2 illustriert. Unter in Normalbedingungen nur schwach virulente Arten (verschiedeneFusarien undPhoma exigua var. exigua) waren deutlich aktiver in kontrollierter Atmosph?re. Dies was besonders der Fall fürF. culmorum. Pilze, die im Versuch unter Normalbedingungen nicht pathogen waren (Colletotrichum coccodes, Cylindrocarpon sp., Phoma eupyrena), haben unter kontrollierter Atmosph?re Sch?den verursacht. Hingegen werden die beiden unter Normalbedingungen sehr pathogenen Arten (F. sulphureum undP., exigua var.foveata), durch kontrollierte Atmosph?re wenig beeinflusst. Es wurde sogar in einigen F?llen eine gewisse Verringerung des F?ulnisbefalls beobachtet. Ganz allgemein haben die getesteten kontrollierten Bedingungen die Entwicklung der Pilzf?ulnis an beiden Kartoffelsorten gef?rdert, speziell durch Pilzarten, welche unter Normal-bedingungen wenig Sch?den verursachen.

     

Marine Pharmacology in 2016–2017: Marine Compounds with Antibacterial,Antidiabetic, Antifungal,Anti-Inflammatory,Antiprotozoal, Antituberculosis and Antiviral Activities; Affecting the Immune and Nervous Systems,and Other Miscellaneous Mechanisms of Action

The review of the 2016–2017 marine pharmacology literature was prepared in a manner similar as the 10 prior reviews of this series. Preclinical marine pharmacology research during 2016–2017 assessed 313 marine compounds with novel pharmacology reported by a growing number of investigators from 54 countries. The peer-reviewed literature reported antibacterial, antifungal, antiprotozoal, antituberculosis, and antiviral activities for 123 marine natural products, 111 marine compounds with antidiabetic and anti-inflammatory activities as well as affecting the immune and nervous system, while in contrast 79 marine compounds displayed miscellaneous mechanisms of action which upon further investigation may contribute to several pharmacological classes. Therefore, in 2016–2017, the preclinical marine natural product pharmacology pipeline generated both novel pharmacology as well as potentially new lead compounds for the growing clinical marine pharmaceutical pipeline, and thus sustained with its contributions the global research for novel and effective therapeutic strategies for multiple disease categories.     

Effects of Micronutrient Deficiencies on Human Health: Its Status in South Asia

SUMMARY

Vitamin A, iodine, and iron deficiencies affect large numbers of people worldwide. A deficiency of one or more of these micronutrients adversely affects the physical and mental abilities of humans. South Asia contains a high percentage of these individuals, not just because of its large population, but also the prevalence of deficiency is higher there than in many other parts of the world. The reasons for the high rates of deficiency in South Asia are many, but they include inadequate availability of micronutrient-rich foods, sub-optimal dietary habits, and high rates of infection. Across the region, national plans vary from (a) increasing availability of appropriate foods, (b) introducing new dietary behaviors, (c) improving health status, and (d) directly addressing deficiency through intervention programs. The extent to which each of the countries is making progress toward the goals of these plans can help in understanding the priorities for future efforts to address micronutrient malnutrition. This article discusses the adverse effects of micronutrient deficiencies in humans with special reference to South Asia.     

稻文化的再思考3:稻与社稷——印度、泰国等东南亚、南亚主要国家

东南亚、南亚地区是世界上重要的稻米产区,稻米在民众饮食生活和国家社会政治中占有重要地位,民众生活中也保存了非常悠久和丰富的稻作文化习俗及稻谷起源神话传说。本文分析了水稻在东南亚、南亚主要国家印度、印度尼西亚、泰国、越南、菲律宾、孟加拉国、缅甸的作用、地位、生产、消费、文化等情况,并指出了印度、印度尼西亚稻文化的主要标志。     

Irrigation but not N fertilization enhances seedhead density in plains rough fescue ( F estuca hallii)

Plains rough fescue ( F estuca hallii) is an important forage grass species in western Canada. Seed for use in pastures and ecological restoration is in high demand but supply is limited because F. hallii is an erratic seed producer. Seed producers require an understanding of the factors that influence flowering and seed set in this species. The aim of this study was to assess the effect of irrigation and nitrogen fertilization on the flowering and growth of F. hallii in a native rough fescue prairie in Alberta, Canada. Irrigation had a strong positive effect on seedhead density, whereas fertilization had limited effects on growth and reproduction of F. hallii. These results demonstrate that under field conditions, available moisture is likely to be a key factor driving flowering and seed production in F. hallii.     

Management of Soil Micronutrient Deficiencies in the Rice-Wheat Cropping System

Summary

Within the last three decades, the rice-wheat cropping system has triggered, and with time, aggravated soil micronutrient deficiencies in the Indo-Gangetic Plains (IGP). This has largely been due to the shift from an earlier rice and wheat monoculture with low yielding, long duration indigenous varieties, to an intensive rice-wheat rotation cropping system with short duration modern high yielding varieties on the same piece of land. The problems related to micronutrient deficiency in the IGP are more due to the size of its available pools in the soil rather than its total contents and are greatly influenced by crop management, or rather its mismanagement. Deficiency of zinc is widespread in the IGP, but with the extensive use of zinc sulfate, zinc deficiency has reduced in some areas of the region. Meanwhile, the deficiency of Fe, Mn and B has increased in the IGP. Deficiency of Cu and Mo is location specific and can limit rice and wheat yields. The adoption and spread of the rice-wheat system in permeable coarse textured soils, particularly in the western IGP, not only caused iron deficiency in rice but also resulted in the emergence of manganese deficiency in wheat. In highly calcareous and acidic soils, boron is the next limiting micronutrient in crop production after zinc. Bumper rice and wheat harvests in the past decade, the declining use of organic manures in the region and except for the widespread use of zinc sulfate, a general lack of awareness amongst farmers on micronutrient deficiency problems has contributed to micronutrients limiting rice and wheat yields in the IGP. Approaches to alleviating micronutrient deficiencies include matching the crop removals of the micronutrients with its replenishments through their respective external carriers, supplementation through organic sources and mobilization/utilization through cultivation of micronutrient efficient crop cultivars. Identification of efficient micronutrient carriers and finding the optimum rate, mode and time of its application is important in ameliorating the micronutrient deficiencies. This article reviews the extent of micronutrient deficiency and discusses various management options available to reduce micronutrient deficiency induced crop yield reduction for rice and wheat in the Indo-Gangetic Plains.     

Model of wheat yield response to application of diclofop-methyl to control ryegrass (Lolium rigidum)

A general model of crop yield response to herbicide application is proposed. The model includes three components: the effect of herbicide dosage on weed density, the effect of surviving weed density on crop yield and the effect of herbicide directly on the crop. The model is used to estimate the response of wheat yield to application of diclofop-methyl to control ryegrass (Lolium rigidum) in Australia. It is found that the competitiveness of ryegrass plants surviving treatment is reduced by the treatment and that the proportion of yield loss at a given ryegrass density is not independent of the absolute weed-free yield. The response function is used to calculate economic thresholds and optimal herbicide dosages.