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Study on Changes in thiol protease activity during germination and its purification

¹Dr. Krishan Pal and ²Ankita Devi

¹Associate Professor, Department of Botany, OPJS University, Churu, Rajasthan (India)

²Research Scholar, Department of Botanu, OPJS University, Churu, Rajasthan (India)

e-mail: ankita39999panwar@gmail.com

Abstract: Thiol protease activity during germination of mungbean seeds was monitored for 5 days using BANA as substrate to determine the optimum stage to harvest seeds for purification of this enzyme. The activity of thiol protease increased markedly after the first day of imbibition, reaching a peak after 2 days and then declining gradually to 15% of the maximum after 5 days. In dry seeds (0 day after imbibition), the enzyme showed 73% of the maximum activity. Thus, 2-day old germinated seeds were used for purification of the enzyme. After ammonium sulphate fractionation the overall fold purification was 12.08 with a recovery of 67 %. The profile of the enzyme elution through DEAE Sephadex A-50 column. The use of anion-exchange chromatography at pH 6.8 separated Leu-βNA hydrolyzing activity (Leucine aminopeptidase activity) and some unwanted proteins from the enzyme. The adsorbed BANA hydrolyzing (thiol protease) activity eluted at 0.3 M NaCl whereas Leu-βNA-hydrolyzing activity was eluted at 0.38 M NaCl. The partially purified fraction obtained from anion exchange column after dialysis, was chromatographed on Sephadex G-100 column. Lots of contaminating proteins were removed at this step and a single peak of enzyme activity was obtained. The enzymatically active fractions were pooled and the pooled fraction when analyzed by native PAGE did not give a single protein band. The next step of purification was affinity column chromatography which was so designed as to separate all the non-thiol proteins from the thiol protease. The affinity column was prepared by immobilizing p-aminophenylmercuric acetate on CNBr-activated Sepharose 4B. The bound enzyme was eluted from the organomercurial column with 50 mM sodium acetate buffer, pH 5.5 containing 0.2M NaCl, 1mM EDTA and 10mM cysteine. The affinity chromatography showed a single peak of enzyme activity. The fractions containing enzyme activity were pooled and immediately dialyzed.

[Dr. Krishan Pal and Ankita Devi. Study on Changes in thiol protease activity during germination and its purification. Academ Arena 2019;11(11):1-5]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 1. doi:10.7537/marsaaj111119.01.

Keywords: Thiol protease activity, Germination, Purification, Mungbean.

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我对太极图理解和运用 （新版）

孙纯武 (Sun Chunwu)

Xiuxiu Garden, Xihu Town, Yangzhou City, Jiangsu Province, China 5008 Mainly engaged in the research of natural sciences, etc.;

Name of the unit, Yangzhou Huading Electric, Jiangsu Province

Set Email: yzscw0514@163.com

Abstract: Knowing the material basis of the original celestial body, it is the force of the force to push forward the frictional motion. Its principle mechanism is to transform the sputum and copy more energy or force. The celestial body is from simple to complex. From the low-level to the advanced step-by-step evolution, the universe of all things or celestial bodies who are of great quality or proper method, who is a unified field in this living field. There is no immutability. They make a one-dimensional repetitive and orderly movement in time, and space is the orderly movement of disorderly movement, irregular velocity, irregularity, and asymmetrical elliptical motion. Explain that the universe's celestial bodies should start with the perfection of physical theory to solve the problem. Solved the physics theory, all the breakthrough problems have been solved. It is known that if a star moves upwards as one of the stars in the center of the galaxy, it will be consumed by the friction force on the field, and the inner core of the planet will be copied. The rotation force is reduced, and the rotation speed is slowed down. Therefore, a planet moving upwards forms a density wave when it is elliptical. The light transformed by the friction on the planet is also darkened, and it is shot at the center of the galaxy. And another ice age in winter.

[孙纯武 (Sun Chunwu). 我对太极图理解和运用 （新版）. Academ Arena 2019;11(11):6-19]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 2. doi:10.7537/marsaaj111119.02.

Keywords: 关键词: 天体; 物质; 能量; 力; 偏心涡旋发动机; 星球; 星系; 中心; 作向; 心涡旋环绕; 椭圆运动; 密度波

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Heavy Metal Contents of Cassava (Manihot esculenta) Leaves obtained from Andoni Local Government Area of Rivers State

Victoria Daminabo and Susan Okparanta

Department of Science Laboratory Technology, School of Science and Technology, Captain Elechi Amadi Polytechnic, Rumuola, P.M.B. 5936, Port Harcourt, Rivers State, Nigeria

vicjodish@yahoo.co.uk

Abstract: The concentrations of selected heavy metals in cassava leaves obtained from Andoni Local Government Area of Rivers State, Nigeria were determined using Atomic Absorption Spectrophotometer (AAS) and compared with the soils on which the cassava grows. Results obtained showed high level of heavy metals content in the cassava. Results indicated the order; zinc (Zn) > lead (Pb) > mercury (Hg) in the cassava leaves when compared with heavy metals detected in the different control soils on which they were grown. For all the farms, the soil plant transfer values were in the order; Pb> Zn> Hg. This shows that among the three metals studied, lead was the easiest to migrate followed by zinc and then, mercury. This trend signifies that plant absorbs a higher concentration of lead from the soil in which it grows when compared to other metals absorbed while Hg is the least absorbed metal. It was observed that the lead content in cassava leaves was above the World Health Organization (WHO)/Food and Agricultural Organization (FAO) recommended safe limit.

[Daminabo, V. and Okparanta, S. Heavy Metal Contents of Cassava (Manihot esculenta) Leaves obtained from Andoni Local Government Area of Rivers State. Academ Arena 2019;11(11):20-29]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 3. doi:10.7537/marsaaj111119.03.

Keywords: Cassava leaves, soil, heavy metals, farms, World Health Organization, safe limit.

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Personal Parameters Related Behavioural Study Of Life Insurance Customers In District Karnal, Haryana (India)

Dr. Gulab Singh Parmar and Ms. Nutan

Assistant Professor, Department of Commerce, OPJS University, Churu, Rajasthan (India)

Research Scholar, Department of Commerce, OPJS University, Churu, Rajasthan (India)

e-mail-nutanahlawat.amity2009@gmail.com

Abstract: In the present study customer level of LIC. As per response 45% response cover from services where as 27% response from profession. It means LIC customers/consumers mostly covers from service sector which is 45% of total survey out minimum from progression that is 27% from the overall study. It means LIC should devote more effects to cover businessman as can as professional customer so that LIC well stay in acute competitions. Age group of customers. As per response 11% customers belong to 10-20 years. 31% customers belong to 20-30 years, irrespective of their occupational level. 43% customers belong to 30-40 years which cover services and business occupation. 30% customer belong to above 40 years. Income group of customers. As per survey 9% customer category are below 50000, whereas 29% customer are related to 50000-1lakh income group. Only 5% customer category of above 3 lakh 37% customer are related to 1 lakh to 2 lakh and 20% customer are related to 2 lakh-3 lakh. It was found high income group respondent are adopted maximum amount policy as compare low income group. Because their financial position will be strong.

[Gulab Singh Parmar and Ms. Nutan. Personal Parameters Related Behavioural Study Of Life Insurance Customers In District Karnal, Haryana (India). Academ Arena 2019;11(11):30-34]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 4. doi:10.7537/marsaaj111119.04.

Keywords: Respondents, Behavioural Study. Life Insurance Company, Karnal, Haryana.

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维京人与“天下人”世界比拼科学解读 ----嫘祖诞生地还有争议的最后确证原理

天文杨

Recommended: 王德奎 (Wang Dekui), y-tx@163.com

Abstract: 摘要：要解决类似李进副省长提出的“嫘祖诞生地除绵阳市盐亭县外，国内其它省市也还有争诞生地的”难题，面对国内国外的各自为阵，说明不是一批专家单纯用史料和文物能决定下来的。本文借用“维京人与‘天下人’世界比拼”来科学解读，是几十年间笔者与各派专家交流的最新感受。

[天文杨. 维京人与“天下人”世界比拼科学解读 ----嫘祖诞生地还有争议的最后确证原理. Academ Arena 2019;11(11):35-110]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 5. doi:10.7537/marsaaj111119.05.

Keywords 关键词：科商 青藏高原 维京人 天下人 摇钱树

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史前考古已发现2000处遗址为啥还尴尬----四川宽窄科学研究之二

桂韩李

Recommended: 王德奎 (Wang Dekui), y-tx@163.com

Abstract: 摘要：2019年8月28日由中国历史研究院、中国考古学会主办的第二届中国考古·郑州论坛，主题为“新中国考古70年”，300余位国内高校、科研单位的专家学者相聚郑州，探讨考古学术热点问题，宣传考古成果，促进学术交流。中科院古脊椎动物与古人类研究所研究员、中国考古学会旧石器考古专业委员会主任高星教授，代表旧石器考古专业委员会总结新中国成立70年来，旧石器考古领域取得的成果，他在会上介绍说：“目前，我国已发现旧石器时代遗址逾2000处，涵盖距今210万年至1万年的各个时段，已构筑了华夏大地史前历史的基础和基干……目前考古存在的问题是学科定位与归属尴尬”。

[桂韩李. 史前考古已发现2000处遗址为啥还尴尬----四川宽窄科学研究之二. Academ Arena 2019;11(11):111-118]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 6. doi:10.7537/marsaaj111119.06.

Keywords: 关键词：史前; 考古; 发现; 遗址; 尴尬

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向宝云院长与杜钢建教授的宽窄之争 ----四川宽窄科学研究之四

天文杨

Recommended: 王德奎 (Wang Dekui), y-tx@163.com

Abstract: 摘要：读2019年9月9日“大同思想网”上杜钢建教授的《“英人英语源于中国”热议的声明》，很惊异。他说：“‘英国起源于湖南’、‘英国人和英语起源于湖南’的说法，是以四川社科院向院长为代表的一些不好学习又喜欢贬低别人的领导们和打手们原创的。四川社科院院长向宝云的原话是‘英国人和英语起源于湖南’，然后发表到各大媒体后变成标题党‘英国起源于湖南’，想必这得到了向院长的认可，或者就是记者与向院长的合谋，也就是说这两句话的发明人是向院长。本人说过‘英国英语起源于大湘西’，但绝不是‘英国起源于湖南’，且不说这个观点是否有道理，这句话本身就逻辑不通，莫非古英国作为一个国家可以从湖南搬到大不列颠岛，或从湖南一路扩张到欧洲？符合逻辑的说法，应是英国人起源于大湘西，即古英国人从中国南方一步步迁移到欧洲。向院长口口声声说要治学严谨，可是这里体现的是却是堂堂一个院长的轻薄和毫不严谨。向院长发明了‘英国人和英语起源于湖南’、‘英国起源于湖南’，然后又把这些个帽子扣到我头上，以逞口舌之快。网络上，还有一大批人以我的名义造假，对我进行污蔑、人身攻击。有一股向院长们的势力，在努力将我‘沦为笑谈’，实实在在体现了学界和媒体界的轻浮，和这个时代的荒谬……对于这些恶意笑谈的始作俑者，我更不愿意忍人之恶”。

[天文杨. 向宝云院长与杜钢建教授的宽窄之争 ----四川宽窄科学研究之四. Academ Arena 2019;11(11):119-127]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 7. doi:10.7537/marsaaj111119.07.

Keywords: 关键词：向宝云, 院长, 杜钢建, 教授, 宽窄

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张军平教授用宽窄论时商----四川宽窄科学研究之六

张军平（复旦大学教授）

Recommended: 王德奎 (Wang Dekui), y-tx@163.com

Abstract: 摘要：2019年7月四川嫘祖蚕丝绸文化研究一带一路《维京人与“天下人”世界比拼科学解读》提出“科商”概念后，9月复旦大学教授、博士生导师张军平先生又提出“时商”概念。“科商”+“时商”是一种时代的必然。张军平教授说：熟能生巧，“熟读唐诗三百诗，不会吟诗也会吟”----这就是提高“时商”的一种办法。这里说学习要努力，要下功夫，其实这也指如何提高“科商”的办法之一。

[张军平（复旦大学教授）. 张军平教授用宽窄论时商----四川宽窄科学研究之六. Academ Arena 2019;11(11):128-129]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 8. doi:10.7537/marsaaj111119.08.

Key words: 张军平; 教授; 宽窄论; 时商

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人文殊途同归的科学前沿《中医药多体自然叩问》前言----四川宽窄科学研究之七

王德奎, 林艺彬, 孙双喜

y-tx@163.com

Abstract: 摘要：知识能改变人的命运；知识能给人力量----那么人对知识的阅读、学习、研究，能提高人自身的身体素质，保持人的身心健康吗？这就是《中医药多体自然叩问》一书想作的试验和探索。我们是一批中医药爱好者，数十年来坚持不懈地对前沿科技知识的阅读、学习和进行研究，亲身的实践确实体会到人文殊途同归的科学前沿，对提高自己的身体素质，保持自己的身心健康有补益。特总结了近数十年，我们在阅读、学习和研究前沿科技方面，叩问地球、叩问宇宙、叩问量子等数十篇心得体会的笔记文章，抛砖引玉献给读者，以求共同进步。

[王德奎 林艺彬 孙双喜. 人文殊途同归的科学前沿《中医药多体自然叩问》前言----四川宽窄科学研究之七. Academ Arena 2019;11(11):130-132]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 9. doi:10.7537/marsaaj111119.09.

Keywords: 关键词: 中医药; 多体; 自然; 知识; 命运；人力

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彭雷用宽窄论人工智能及医药与展开----四川宽窄科学研究之九

彭雷（“客如云”首席执行官（CEO））

Recommended: 王德奎 (Wang Dekui), y-tx@163.com

Abstract: 摘要：2019年10月22日“科学网” 彭雷博客专栏，“客如云”创始人彭雷CEO（首席执行官）发表论“人工智能宽窄要求新时代人才”的文章，有导向意义，特摘录转发----2019年10月20日第六届世界互联网大会在浙江乌镇开幕，10月22日“新浪财经”网发“ 电科技”报道 ----又到一年的乌镇互联网大会，如往年一样，业界大佬悉数到场，就互联网产业经济分享自己的看法，共同探讨未来数字经济的发展走向。和往年不一样的是，今年的乌镇没有了推杯换盏的饭局，但竞争不会因此而停歇，站队不会消失，硝烟依然弥漫。彭雷，1980年生，四川成都人，1998– 2003年获中国科技大学学士学位，2003– 2005年获香港理工大学硕士学位。从大学开始至今彭雷十余年创立了多个涵盖不同领域的公司，包括：O2O、团购、数据库营销、数据挖掘、CRM、SNS到企业应用等。彭雷2003年首创医疗行业信息费用监控管理系统。2006年创立了名片网。2007年创立了FirstDM营销服务数据库。2010年创立了24券团购网并出任COO。2012年彭雷从24券离职任移动互联网创新企业客如云创始人兼CEO。客如云通过商户端硬件及应用、电话、短信、APP、电子邮件等多种方式，结合移动互联网时代云科技的特质进行颠覆性改革，让亿万商家和顾客同步进入移动互联网云时代。

[彭雷. 彭雷用宽窄论人工智能及医药与展开----四川宽窄科学研究之九. Academ Arena 2019;11(11):133-135]. ISSN 1553-992X (print); ISSN 2158-771X (online). http://www.sciencepub.net/academia. 10. doi:10.7537/marsaaj111119.10.

Keywords: 关键词：彭雷; 宽窄论; 人工智能; 医药

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