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Anti-allergic effects of ethanol extracts from brown seaweeds

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Journal of Zhejiang University-Science B(Biomedicine & Biotechnology) 2009年第2期10卷 147-153页

作者： haider samee Zhen-xing LI Hong LIN Jamil KHALID Yong-chao GUO Food Safety Laboratory College of Food Science and Engineering Ocean University of China Qingdao 266003 China Food and Marine Resources Research Centre Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex Karachi 75280 Pakistan

Ethanol extracts of brown seaweeds from Pakistan and China were isolated and compared for their antiallergenic activities. They included Sargassum tennerimum (ST) and Sargassum cervicorne (SC) from Pakistan, and Sargassum gramini- folium turn (SG), Sargassum thunbergii (STH), and Laminaria japonica (LJ) from China. The ethanol extracts of these brown seaweeds were optimized at 85% (v/v) ethanol for the maximum yield of phlorotannin, an inhibitor against hyaluronidase. Total phlorotannins contained in the crude extracts were measured as 1.71% (SG), 0.74% (STH), 0.97% (LJ), 3.30% (SC), and 5.06% (ST). The 50% inhibitory concentrations (IC50) of Pakistani SC and ST were 109.5 and 21 μg/ml, respectively, lower than those of Chinese SG, STH, and LJ (134, 269, and 148 μg/ml, respectively). An antiallergic drug, disodium cromoglycate (DSCG), had an IC50=39 μg/ml, and a natural inhibitor of hyaluronidase, catechin, had an IC50=20 μg/ml. The IC50 of ST extract was found similar to that of catechin (21 vs 20 μg/ml) and lower than that of DSCG (21 vs 39 μg/ml). This suggests that ST is a potent inhibitor of hyaluronidase, indicating a promising future development of natural antiallergic medicines or functional foods.

关键词： Anti-allergic activity Brown seaweed Ethanol extracts Hyaluronidase Phlorotannin

Optimization of Preparative Separation and Purification of Total Polyphenols from Sargassum tenerrimum by Column Chromatography

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Journal of Ocean University of China 2009年第4期8卷 425-430页

作者： samee haider LI Zhenxing LIN Hong Khalid Jamil Food Safety Laboratory College of Food Science and Engineering Ocean University of China Qingdao 266003 P. R. China Food and Marine Resources Research Centre PCSIR Laboratories Complex Off University Road Karachi Pakistan

Polyphenols from the ethanol extracts of Sargassum tenerrimum （ST） with potent antiallergic effects were studied to optimize separation process through column chromatography. The adsorption and desorption characteristics of three widely used adsorbents： macroporous resin, silica gel, and polyvinylpolypyrrolidone （PVPP）, were critically evaluated respectively and studied for the optimization of preparative separation of polyphenols. Static operations on these adsorbents showed that macroporous resin had the best adsorption and desorption capability among the three adsorbents. Dynamic adsorption and desorption with macroporous resin packed column were also conducted to optimize the parameters such as： with the optimal values shown in brackets, the concen- tration of extract solution （4 times diluted）, pH value （6-7）, adsorption speed （3BVh-1, bed volumes/per hour）, concentration of ethanol （80%）, elution speed （3 BV ht） and elution volume （7 BV）. The chromatographic process so optimized gave a purity of 62.43% from the crude polyphenols, providing a promising basis for large scale preparation of bioactive polyphenols upon further scaling up tests.

关键词： Sargassum tenerrimum polyphenol separation adsorption desorption column chromatography

Studies on the Specific Degranulation of Mast Cell Sensitized by Several Allergens in vitro

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Cellular & Molecular Immunology 2009年第2期6卷 149-153页

作者： Yongchao Guo Zhenxing Li Hong Lin haider samee Jamil Khalid Food safety laboratory Ocean University of China Qingdao 266003. Shandong China Pakistan Council for Scientific and Industrial Research Laboratories Complex Karachi Pakistan

Food allergy is a major health issue worldwide. Mast cells play a very important role in the immediate hypersensitivity for which mast cell degranulation needs to be studied extensively. In this study, an approach was taken to study the characteristics of sensitized mast cell degranulation in vitro, which associated with the study of mast cells and animal models. BALB/c mice were immunized respectively by several food allergens, then blood and peritoneal mast cells were collected at different time points. A dynamic determination was carried out between mast cells and serumal IgE. Comparative analysis on sequential time points showed that there was a close coincidence between mast cell degranulation and IgE antibody titers in sensitized BALB/c mice. Furthermore, it is interesting that sensitized mast cells could implement specific degranulation against the challenges in vitro, but the closely tropomyosins induced mast cell degranulation displayed cross reactions. This is very similar to IgE resisting the allergens in vivo. The study disclosed some characteristics on mast cells, coming from sensitized BALB/c mice, degranulation in vitro.

关键词： food allergy allergen mast cell degranulation

刀额新对虾主要过敏原蛋白的肽指纹图谱鉴定与分析

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中国海洋大学学报（自然科学版） 2009年第5期39卷 919-924页

作者：林洪郑礼娜李振兴 samee haider 中国海洋大学水产品安全实验室山东青岛266003

以我国常见的刀额新对虾为研究对象,旨在获得其主要过敏原的氨基酸序列等基础数据,增加对虾类过敏原的认识和理解。分离和纯化了分子量为36 kD的虾主要过敏原蛋白,利用激光辅助解析/飞行时间质谱对其进行肽质量指纹图谱鉴定,并对鉴定的... 详细信息

以我国常见的刀额新对虾为研究对象,旨在获得其主要过敏原的氨基酸序列等基础数据,增加对虾类过敏原的认识和理解。分离和纯化了分子量为36 kD的虾主要过敏原蛋白,利用激光辅助解析/飞行时间质谱对其进行肽质量指纹图谱鉴定,并对鉴定的结果采用Mascot搜索引擎在NCBInr数据库上进行搜索。结果表明刀额新对虾主要过敏原蛋白与斑节对虾原肌球蛋白匹配分值最高为268,吻合肽段27条,序列覆盖率为65%;与其它无脊椎动物如腐食酪螨、衣鱼等的原肌球蛋白的序列覆盖率也很高,分别达到了51%和53%。这一结果不仅表明了刀额新对虾与其它甲壳类海产品过敏原存在着高度同源性,而且为其与甲壳类及其它无脊椎动物主要过敏原之间存在严重交叉反应的现象提供了理论依据。

关键词：刀额新对虾过敏原激光辅助解析/飞行时间质谱鉴定

菲律宾蛤仔过敏原可视化抗体微阵列玻片的检测方法

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水产学报 2010年第3期34卷 422-427页

作者：李小燕李振兴林洪 samee haider 张轶群中国海洋大学水产品安全性实验室 Food and Marine Research Center PCSIR Laboratories ComplexOff University Road

采用辣根过氧化物酶(HRP)与3,3′,5,5′—四甲基联苯胺(TMB)—H2O2作为信号示踪系统,建立了一种可视化抗体微阵列检测菲律宾蛤仔过敏原的方法。对孵育时间、抗体浓度等免疫条件进行了优化。采用改良双抗体夹心法,将菲律宾蛤仔过敏原的... 详细信息

采用辣根过氧化物酶(HRP)与3,3′,5,5′—四甲基联苯胺(TMB)—H2O2作为信号示踪系统,建立了一种可视化抗体微阵列检测菲律宾蛤仔过敏原的方法。对孵育时间、抗体浓度等免疫条件进行了优化。采用改良双抗体夹心法,将菲律宾蛤仔过敏原的兔多克隆抗体固定于琼脂糖三维芯片上,依次加入待检样品、菲律宾蛤仔鼠多克隆抗体和HRP-羊抗鼠IgG,孵育后加TMB显色,肉眼观察后,用平板扫描仪获取扫描图象,采用GenePixPro6.0软件分析灰度值。试验结果表明,该方法最低可检出10ng/mL的菲律宾蛤仔过敏原,片内平均变异系数(CV)为5.99%,片间为10.3%;香肠及蟹棒中3个不同浓度的加标回收率为73.54%～95.44%;4℃存放4个月内抗体微阵列玻片活性保持稳定。该方法不需要大型精密仪器,结果直观可见,可以发展为对多种过敏原进行同时检测,具有良好的实用和推广价值。

关键词：菲律宾蛤仔过敏原夹心法抗体微阵列可视化