Carrefour's Misadventure in Russia Case Study Example | Topics and Well Written Essays - 2000 words

Carrefour's Misadventure in Russia - Case Study Example This research will begin with the statement that with a move to the globalized era, successful businesses and large corporations are continuously looking for the opportunities of international expansion. The last trends show that emerging economies, especially BRIC (Brazil, Russia, India, and China) become more popular destinations for business development purposes. While there are many examples of successful penetration and business establishment on these markets, there are also examples of failures and misadventures. Entry strategy of the France-based retail giant Carrefour to the Russian market has been unsuccessful, and the company’s sudden exit has raised many different opinions in this respect.   Below there is provided an overview of potential factors that might have its impact on the company’s failure continued with the further analysis of the Russian food retail market based on the Porters’ five forces model. While there was not clearly defined the rea son of why Carrefour has made a decision to exit suddenly the Russian market, there were mentioned some obvious factors that might have contributed to this decision. These factors include: social and cultural factors; legal factors; late entry decision; absence of sufficient organic-growth prospects and acquisition opportunities, and political factors. Such social and cultural factors that have been identified as the potential barriers to the Carrefour’s expansion in Russia included: bureaucracy, corruption, red tape.

Formula Milk Essay Example for Free

Formula Milk Essay Annual international trade of barley averages around 15 million metric ton and fluctuates from 12 to 16 million metric tons. This is about 9 -12% of total global production. Major exporters are EU, Australia, Ukraine, Canada and UK. US is neither a major exporter nor a major importer. Its export and imports are more or less same. US export mainly the feed barley and imports malt barley. The annual trade of feed barley comprises more than 75% of total barley trade while that of malt barley accounts for less than 25%. China and US being leaders in beer production account for over 60% of malt barley trade. China alone imports 50% while US about 11%. Leading Barley Import and Export Countries (Averages for 1998-2000) Country Imports (Mt x 1,000) Country Exports (Mt x 1,000) Saudi Arabia 4447 France 6758 Japan 2531 Germany 4040 China 2160 Australia 3973 Belgium-Luxembourg 1459 Canada 2196 Russian Federation 1005 United Kingdom 1898 Germany 941 Belgium-Luxembourg 1223 Brazil 880 United States 985 Netherlands 823 Denmark 960 Morocco 794 Ukraine 841. Italy 790 Turkey 659 United States 778 Sweden 606 Table 4: Global trade of barley in 1998-2000 (faostat) Sourcing Barley for Beer Production Malting companies in US processes about 3 million ton of malt barley annually of which about 1. 0 million ton is imported barley and the remaining (about 2 million ton) is produced in US itself. Therefore, a beer company in US should keep both the options – of sourcing from within the country from states like North Dakota, Montana and Idaho etc. and of importing open for sourcing barley. When it comes to import Canada should be the obvious choice due to geographic proximity. The next obvious choice has to be EU. As far as sourcing from domestic market is concerned the company can have a direct contract with major barley growers to save cost and ensure a reliable supply. In case of import from Canada the concerned agency is Canadian Wheat Board. Concerning Outsourcing of the product, a beer company should focus on Malting, Brewing, and Fermentation etc. instead of imports and domestic purchasing of commodities. Therefore, it is always better to outsource the product to a few good commodity trading companies. The commodity should be outsourced not to one company rather to more than one company to have a bargaining power over pricing and also to ensure a uninterrupted supply of barley in case of any unforeseen crisis to keep the brewery running. There can be another outsourcing approach for a new start up Beer Company. With malting being an established process. There are dozens of malting companies in US and hundreds on the globe and malt being a standard product being traded in the market globally. Therefore, for a start up Beer Company it will be a good idea to focus on brewing and fermentation, rather than going for setting up the malting facility. If the company specializes on producing better and special quality beer it can command better price for its beer and that business model should be more profitable than starting fro malting. Some important malt barley and malt suppliers in US are – 1. North Dakota Barley Council 2.Americas Malt, PO Box 5724 Minneapolis, MN 55440-5724 Tel: 1-952-742-5646 Fax: 1-952-742-5050 Internet: www. Cargill. com. Products: Barley Malt 3. Busch Agricultural Resources, Incorporated P. O. Box 427 West Fargo, ND 58078 Tel: 1-701-282-5752 Fax: 1-701-282-6260 Products: Malting Barley 4. Cargill, Inc. P. O. Box 9300 Minneapolis, MN 55440-9300 Tel: 1-952-736-8664 Fax: 1-952-742-6252 Internet: www. Cargill. com Products: Feed Barley, Malting Barley 5. Cenex Harvest States P. O. Box 64089 St. Paul, MN 55164-0089 Tel: 1-651-306-6156 Fax: 1-651-306-6570 Internet: www. harveststates. com Products: Feed Barley, Malting Barley 6. Columbia Grain 111 S. W. Columbia St, Suite 1200 Portland, OR 97201. Tel: 1-503-224-8624 Fax: 1-503-241-0296 Products: Feed Barley, Malting Barley 7. ConAgra Grain Companies 400 4th Street, Suite 850 PO Box 15083 Minneapolis, MN 55415 Tel: 1-612-341-2326 Fax: 1-612-341-2137 Products: Feed Barley, Malting Barley 8. ConAgra Malt P. O. Box 1529 Vancouver, WA 98668-1529 Tel: 1-360-699-9389 Fax: 1-360-696-8354 Products: Barley Malt 9. Coors Brewing Company 17755 West 32 Av. Brewing Materials BC 610 Golden, Colorado 80401 USA Tel:1-208-678-3586 Fax : 1-208-678-9669 Products: Malting Barley, Barley Malt 10. General Mills Operations, Inc. P. O. Box 5022 #2 Fifth Street N. , Suite 200 Great Falls, MT 59401 Tel: 1-406-761-6252 Fax: 1-406-727-8096 Internet: www. gmigrain. com Products: Feed Barley, Malting Barley 10. International Malting Company P. O. Box 712 Milwaukee, WI 53201-0712 Tel: 1-414-649-0206 Fax: 1-414-671-1385 Products: Barley Malt 11. Louis Dreyfus Corporation 222 S. W. Columbia, Suite 1133 Portland, OR 97201 Tel: 1-503-243-1133 Fax: 1-503-243-5079 Products: Feed Barley 12. Rahr Malt 800 West First Avenue Shakopee, MN 55379 Tel: 1-952-496-7016 Fax: 1-952-496-7054 Products: Barley Malt. 13. United Harvest 200 S. W. Market Street, Suite 1780 Portland, OR 97201 Tel: 1-503-344-1900 Fax: 1-503-944-1969 Internet: www. harveststates. com Products: Feed Barley, Malting Barley Some International suppliers of malt and malted barley are listed below. 1. Muntons plc, Cedars Maltings Stowmarket, Suffolk IP14 2AG, UK t. (+44) (0) 1449 618300 f. (+44) (0) 1449 677800 [emailprotected] com 2. Weyermann Specialty Malting Company Brennerstrasse 17-19 96052 Bamberg, Germany Phone: + 49 (0)951 93 220-12 Fax + 49 (0)951 93 220-912 eMail [emailprotected] de 3. Agniche Marketing Ltd. 70 Johnson, Birtle, MB, Canada, R0M 0C0 Telephone: 1-204-842-5132 Fax: 1-204-842-3778 4. Bairds Malt Limited Elliot Industrial Estate, Arbroath, Angus, DD11 2NJ Scotland Tel: +44 (0)1241 870431 Fax: +44 (0)1241 874251 www. bairds-malt. co. uk E-mail: [emailprotected] co. uk The price of domestic malt barley in US is around 250$/metric ton, while the price of malt barley imported from EU is about 280$/metric ton. For exact price the company will seek quotations from a couple of potential domestic as well as international suppliers. After examining the quotations 4-5 potential suppliers will be selected for further price negotiations to seek the minimum possible price. The supplier will be selected on the basis of past record for reliability, the cost and the quality of the barley. Supply contract should be made with 2-3 suppliers to have secure supply of the raw material for beer production. 8. References 1. http://en. wikipedia. org/wiki/Barley#_note-1 2. http://faostat. fao. org 3. http://www. fapri. missouri. edu 4. http://www. ambainc. org/ni/index. htm.

uss indianapolis :: essays research papers

USS Indianapolis The USS Indianapolis (CA-35) was commissioned at the Philadelphia Navy Yard on 15 November 1932. The ship served with honor from Pearl Harbor through the last campaign of World War II, sinking in action two weeks before the end of the war. On 30 July 1945, while sailing from Guam to Leyte, Indianapolis was torpedoed by Japanese submarine I-58. The ship capsized and sank in twelve minutes. Survivors were spotted by a patrol aircraft on 2 August. All air and surface units capable of rescue operations were dispatched to the scene at once, and the surrounding waters were thoroughly searched for survivors. Upon completion of the day and night search on 8 August, 316 men were rescued out of the crew of 1,199. We believe we were hit by two torpedoes, one around frame 8 or 10, because the bow was blown off forward around ten. Another one [torpedo] around frame fifty. We believe that they were large torpedoes, that they were running close to the surface, because none of us believe the magazines blew up, that is the only way we can account for the flashes of flame through the ship. He was able to aft on the starboard side, although badly injured, he didn't get to the main engine room, No. 2 engine room, where he found No. 2 engine had lost vacuum and that was shut down. He did talk to somebody in No. 1 engine room. They told him that apparently the main steamline going through the port side of the forward engine room had been knocked loose. They had no steam and asked for instructions. All power all lights were lost forward. The fact that the [torpedo] hits were there, at least we think they were up forward, are borne out by the fact we have almost no Marines who were reported in that section of the ship. We have not a single steward's mate and their compartment was up there and we have very few officers that were in their rooms at the time of the explosion. So we believe all of those people were killed almost instantly. It's inconceivable that you can't get a message off or that we didn't get a message off. However, we do know that we were in the water about 107 hours, therefore, next time we think that we would like to have a positive means, in case we went down, of somebody saying, "They're gone", somebody with us and tell the outside world about it. uss indianapolis :: essays research papers USS Indianapolis The USS Indianapolis (CA-35) was commissioned at the Philadelphia Navy Yard on 15 November 1932. The ship served with honor from Pearl Harbor through the last campaign of World War II, sinking in action two weeks before the end of the war. On 30 July 1945, while sailing from Guam to Leyte, Indianapolis was torpedoed by Japanese submarine I-58. The ship capsized and sank in twelve minutes. Survivors were spotted by a patrol aircraft on 2 August. All air and surface units capable of rescue operations were dispatched to the scene at once, and the surrounding waters were thoroughly searched for survivors. Upon completion of the day and night search on 8 August, 316 men were rescued out of the crew of 1,199. We believe we were hit by two torpedoes, one around frame 8 or 10, because the bow was blown off forward around ten. Another one [torpedo] around frame fifty. We believe that they were large torpedoes, that they were running close to the surface, because none of us believe the magazines blew up, that is the only way we can account for the flashes of flame through the ship. He was able to aft on the starboard side, although badly injured, he didn't get to the main engine room, No. 2 engine room, where he found No. 2 engine had lost vacuum and that was shut down. He did talk to somebody in No. 1 engine room. They told him that apparently the main steamline going through the port side of the forward engine room had been knocked loose. They had no steam and asked for instructions. All power all lights were lost forward. The fact that the [torpedo] hits were there, at least we think they were up forward, are borne out by the fact we have almost no Marines who were reported in that section of the ship. We have not a single steward's mate and their compartment was up there and we have very few officers that were in their rooms at the time of the explosion. So we believe all of those people were killed almost instantly. It's inconceivable that you can't get a message off or that we didn't get a message off. However, we do know that we were in the water about 107 hours, therefore, next time we think that we would like to have a positive means, in case we went down, of somebody saying, "They're gone", somebody with us and tell the outside world about it.

British Telecommunications and the Drivers of Globalization

Globalization is a phenomenon most often discussed among economists. Most of us have often heard that business is no longer restrained by political or cultural boundaries. Multinational Enterprises (MNEs) expand with little regards of geographical or cultural distance among their target markets. All decisions within a company are made with the additional consideration of how the international world will affect the results of that decision, a factor that most businesses ignored in some earlier decades.As indicated above, discussions about globalization are mostly about its effects toward the business environment, rather than its causes. Companies are more interested toward how a phenomenon will affect their financial conditions rather than where or what the phenomenon is actually coming form. Within this paper however, I will discuss several drivers of globalization. The drivers will be taken from various perspectives about globalization. To provide more connection to the internationa l business environment, we will use the British Telecom, one of the most successful MNEs in the global communication industry, as a reference to help define and justify the drivers of globalization.Furthermore, this paper will discuss the view of whether MNEs are actually competing globally or only regionally. There are arguments among scholars that the global economy do not actually existed. Globalization has taken us through national boundaries but it stopped in regional limits. In other terms, MNEs are not actually operating with global factors within their considerations, but rather regional factors. This view will be examined as we discuss the development of British Telecom.I. Literature ReviewII.1Corporations as Principle DriversAccording to Ron Blackwell (2002), corporations are the principle drivers of globalization. In the process of increasing their market share, corporations found that they need to expand further that their national boundaries allowed. While rapidly inter nationalizing their operations, most corporations fundamentally altered their structure and strategies, boosting the change of their national economy into a more globalize economy.This phenomena might not be observable in the US or European soil, because the process of corporations breaking through national boundaries have occurred several decades ago. However, the phenomenons are still happening today within the Asian and other developing economies. Economist have long predicted that Asia will be the center of massive economic growth in the near future, and the prophecy is fulfilled as China and India are increasingly integrated to the global economy.II.2Political, Technological and Social ConditionsIII.2.1 Market Oriented-PoliciesBeside the principle factor mentioned above, many arguments brought forward other causes of globalization. One of the popular ones is the influence of market-oriented policies. In China for example, the change of governmental policies regarding foreign in vestment and trade was the greatest factor that supported growth of the economy. Several decades ago, China was a country considered to have a ‘closed economy’. Today, the country is the largest recipient of the world’s Foreign Direct Investment and recognized as the most potential market in Asia (Diaz, 2001).III.2.2 Communication and Transportation TechnologyTechnological changes have also contributed to the process of turning the world into a single global economy. The internet spawned the e-commerce, which displayed a tremendous increase in its role in business and trade. As people in modern countries crave the ease of trade provided by the internet, the growth of e-commerce is predicted to last indefinitely. Furthermore, advanced transportation equipment also has a significant role in accelerating the globalization process.II.2.3  Ã‚   Global PhenomenaSocial conditions that supported globalization process consist of several important phenomena. One of those phenomena is the end of the cold war. The reduction of conflicts causes competitive environment for business development and population growth. It was recorded that the number of people on the planet is more than doubled since the 1960’s (Kohl, 2000). Both the reduction of conflicts and the increasing population created more economic, social and environmental linkages, which in the end, spawned globalization. Further analysis and identification of the drivers of globalization will be performed using the British Telecom plc.II. British TelecommunicationsIII.1  Ã‚  Ã‚   Corporate BackgroundBritish Telecommunication Plc is responsible for approximately 25 million telephone lines in the United Kingdom. The company is an international corporation which own and runs most of the telephone exchanges, trunk network and local loop connections for the vast majority of British fixed line telephones. The company, British Telecom is formed in 1981 and has developed itself into five larg e business divisions today, they are: BT Retail, BT Wholesale, Openreach, BT Global Services, BT Exact/One IT. The divisions provide communication services ranging from retailing telecoms to consumers to research & development and consultancy.There are several reasons why the observation on British Telecom is connected to the study of globalization and considered interesting by many. First, the UK telecommunication market was among the first telecommunication market that was deregulated in the early 1980’s. Several of the most influential regulatory innovations, like the price cap regulation was also developed and first implemented in UK telecommunications. Second, the British telecom was the first communication company in the world that was privatized in the early 1980’s. Third, because the UK telecommunication market was the first to be exposed to public interference, communication within the industry is ten years ahead of other European countries.III.2 Corporate Int ernationalizationIII.2.1 Role of Government PolicyAccording to the theories mentioned in the second chapter, there are several drivers of globalization with different emphasize on their significance. Generally, economists acknowledge that companies are the strongest driver of globalization and its processes. Despite the fact that this statement might be true for most MNE’s, regarding the British Telecom, I found that governmental policies are the most influential factor comparatively.Before its privatization in 1984, the British Telecom was granted the right for a monopoly over the fixed line network operations. This boosted the development of the company long enough to provide itself with strong infrastructure to face competitions. Having a major early start at the game, since its privatization, the company has been known as one of the strongest companies in UK. Government monopoly policies which then followed by the market oriented policies helped brought the company to its current position as a market leader in communication (Summanen, 2002).II.2.2  Ã‚   The Role of Corporate ExpansionThe second largest factor, which I believe was the effort within the corporation itself, came in some time after the company was privatized. In 1993, the company announced a joint global alliance through a new joint venture company. This new company is considered as the bridge which leads British Telecom into leading the global telecommunications operator. In order to expand the business by obtaining multinational clients, BT formed and acquired stakes of several joint venture companies overseas. Some of them were Albacom in Italy, Viag Interkom in Germany, Telenordia in Sweden, Dacom in Korea, etc. All of this was performed after there is a leadership change from Sir George Jefferson to Sir Iain Vallance as chairman of the group. This displayed the tremendous role of managerial decisions in the process of BT’s internationalization (Summanen, 2002).II.2.3  Ã‚   Role of Technology and Social FactorsThe changing culture and the increasingly rapid growth of the economy boosted telecommunication business. The 2001residential fixed telephone volumes were recorded to double as much as it was in 1996. Within the same period, local call volumes decreased and national and international call volumes increased. Furthermore, the role of technology in the development of British Telecom can be observed through the statistics also. From 1997 to 2001, calls to mobiles have been recorder to increase tremendously. This increase continues until today (Summanen, 2002).II.2.4  Ã‚   Competing GloballyIn terms of strategic market share, the British Telecom displayed a global approach throughout its lines of business. BT’s overseas activity targets were set to various markets in the world, divided into three segments: the North America market, the Mainland Europe market and the Asia-Pacific region. In North America, the company partnered with MCI to s upport growth. The largest focus is still in its home market and the mainland Europe using its core brands and the market Asia-Pacific region is still being developed (Sumanen, 2002).BibliographyBlackwell, Ron. 2005. ‘Hearing before the US-China Economi ad Security Review Commission Regarding Corporate Globalization Strategies China and the Future of Globalization’.Diaz-Bonilla, E and S Robinson eds. 2001. ‘Shaping Globalization for Poverty Alleviation and Food Security’. 2020 Vision Focus * Washington, D.CKohl, R and K, Orourke. 2000. ‘Whats new About Globalization: Implications for Income Inequality in Developing Countries’ Paper for Organizations for Economic Cooperation and Development Conference on’ Poverty and Income Inequality in Developing Countries’ Paris.Summanen, Tuamo. 2002. ‘British Telecom: Searching for a Winning Strategy’. Available at: msl1.mit.edu/CMI/furd_2002_a/btcase.pdf

Understand the Role of the Nurse

Introduction The purpose of this assignment is to explore the professional responsibilities of the nurse and their role in safeguarding vulnerable patients, all of which are based around a fictitious scenario. The scenario is centred on a patient named Margaret and is going to be used to identify any vulnerability issues Margaret may have. Margaret could be identified as been vulnerable, as she is an 89 year old lady who is suffering from a heart condition. Margaret is currently living with her daughter and has said that her mother’s health has begun to deteriorate.Margaret is becoming very confused and forgetting to take her medication which has resulted in numerous admissions to hospital. No consent was needed on this occasion as it is a fictitious patient. Based around this fictitious scenario, the aim is to discuss and identify vulnerability and any issues arising from it. Exploring all aspects of vulnerability involving all models and discussing confidentiality, looking a t the importance of it and why it should be maintained.This assignment will also look at the mental capacity act and how the nurse and other healthcare professionals assess the patient for capacity and use the models of assessment. All these are in place to protect the patient’s health and well-being and patient rights. Communication is another important skill for any nurse to have. Good communication skills are essential in providing a high standard of care, which will be explored and discussed in detail.It will also explore what accountability and responsibility is within the nurse’s role. The nurse being accountable for their actions and adhere to the Nursing and Midwifery Councils (NMC), Code. â€Å"The core function of the Nursing and Midwifery Council is to establish standards of education, training, conduct and performance for nurses and midwives to ensure those standards are maintained, thereby safeguarding the health and well-being of the public† (NMC, code, 2009).It will then identify and explain how the patient’s autonomy might be promoted by the nurse, highlighting the support available and with the nurse building a therapeutic relationship with the patient, relating to scenario whilst maintaining appropriate professional behaviour and interventions. Overall this assignment will explore and discuss many of the NMC guidelines about the above topics.It will give you an understanding of the nurse’s role involved in delivering the best possible care to the public. Firstly to explore accountability within the nurse’s role The Nursing and Midwifery Council states, â€Å"Accountability is often defined as responsibility, but there is a difference between the two. Responsibility is concerned with answering for what you do, whereas accountability is being answerable for the ‘consequences’ of what you do† (NMC, 2009).

Strong Verbs List Your Detailed Guide to Strong Verbs [Video + Download]

Strong Verbs List Your Detailed Guide to Strong Verbs [Video + Download] Strong Verbs List: Your Detailed Guide to Strong Verbs [Video + Download] Strong verbs are essential for great writing. Not only do you need to know HOW to use powerful verbs, but having a strong verbs list at your disposal is invaluable. Well cover both for you.I used to think writinga book was easy. And in all honesty, writing has never been the most difficult thing in the world for me but when it comes to writing stories and crafting my writing in a way that compels others and pulls them in deeply, it’s been an uphill battle before I discovered using strong verbs along with interesting and cool words to use, that is.Heres what well cover about strong verbs :What is a strong verb?FREE downloadable 200+ strong verbs listStrong verbs versus weak verbsWeak verbs to replace in writingWhat are to be verbsTo be verbs listHow to use strong verbs the right wayWhy you should use strong verbsStrong action verbs for better writingInever struggled with putting my thoughts on paper or even coming up with the ideas.Nope.My biggest hurdle was bringing the emoti on I was trying to convey to life.And as I delved deeper into the literary world, I quickly realized that using strong verbs is a must if you want to create something that leaves a lasting impact. What is a strong verb?Thanks to every English class growing up, you know a verb is a word used to describe an action, state, or occurrence. But what’s the deal with strong verbs? It’s not like they can hit the gym and bulk up, right?A strong verb is a better and more descriptive version of a basic verb that creates a stronger visual and can help create a mood (or vibe) for the scene.Weak VerbStrong Verb VariationsRunBolt, sprint, jogWalkSlink, trot, mopeMakeCreate, forge, foster, brewSaySpeak, voice, purr, mutterSo although you can use the basic verb and still tell the same story, you will create a deeper impact within the reader if you use a strong verb in its place.Strong verbs only help your writingI’ll be real with you here. Using strong verbs will definitely make your writing more intriguing and will increase the chances of someone buying (and loving!) your book.BUT, they won’t help you know how to publish a book that sells.And what’s the point of having incredible writing if you can’t publish it in a way that allows for the world to see it?Well, that’s where we come in. No matter how great your book is, you still need to know how to put it all together, market it, and then self-publish in a way that generates sales.Join Chandler Bolt at his FREE Webinar Training as he reveals the exact tactics and strategies he used to write and publish 6 bestselling books in a row and how he used them to build a 7-figure business in less than 2 years!Spots are limited!Click Here to Save Your SpotStrong verbs VS weak verbsWhat’s the real difference here? How can you tell the difference between a strong verb and a weak one?Since you can’t exactly ask words to flex, you need another system to determine if your verbs are weak or not.Here’s how we define strong verbs vs weak verbs:Weak verbs are the â€Å"basic† forms of a specific action, like â€Å"walked† or â€Å"ran.†Strong verbs are a specified form of a broader action, like â€Å"stomped† or â€Å"bolted.†So the main thing you need to remember when it comes to strong verbs vs weak verbs is how specific it is.HOW TO IMPROVE YOUR WRITING: Use strong verbs instead! Weak verbs are the basic forms of a specific action, like walked or ran. Strong verbs are a specified form of a braoder action, like stomped or bolted.Click To TweetWeak Verbs to Replace in WritingThe weakest verbs you can replace in your writing are to be verbs. These pull your writing quality down and peg you as an amateur.Other weak verbs include basic forms of any verb, like run, walk, say, sit.Now, keep in mind that weak verbs are absolutely okay to use on occasion. The issues arise when youre using these verbs over and over again when there are better, stronger verbs you can use to make your writing more powerful.Sometimes the best verb to use in a situation is the weak verb. Just keep a look out for how often youre using that b asic form so you can beef up your writing by replacing them in other places.What are to be verbs how to replace them in your writing?To be verbs are any verbs used to describe a state of being, including these terms: is, am, are, was were, be, being, been.In order to make your writer stronger, its important when to use them and when theyre making your writing clunky and weak.Here are a few examples of how to replace to be verbs with something stronger:"To Be" Verbs SentenceReplacing "To Be" VerbsReplacing Weak Verb with Strong VerbShe was walking through the corridor.She walked through the corridor.She slinked through the corridor.Conrad is afraid of the dark.Conrad fears the dark.Conrad cowers from the darkness.I was being chased by someone I didn't know.I was chased by someone I didn't know.Someone I didn't know chased after me.I was wanting to visit this forest for as long as I can remember.I wanted to visit this forest for as long as I can remember.I longed to visit this forest for as long as I can remember.To Be Verbs ListOne of the best ways you can strengthen your writing is to use strong to be verbs. These will instantly make your writing more compelling and less amateurish.These are state-of-being verbs like to be verbs to look out for in your writing:AmAreBeBeenBeingCanCouldDidDoDoesHadHasHaveIsMayMightMustShallShouldWasWereWillWouldHow to use strong verbs in writingLittering your writing with strong verbs wont necessarily make it any better. In fact, if you overdo it, those verbs will have the opposite effect.Instead of making your writing stronger, it can bring it down to an amateur level.That being said, I created the video below in order to help you understand how to use strong verbs in your writing the right way. Why use strong verbs for writingBecause your writing will be better overall. One of the best ways you can immediately make our writing stronger is by going through and crossing out each weak verb and replacing it with a better one.Here ’s how your writing will improve when you choose to use strong verbs.#1 Stronger visualsOne of the most important parts of any book is that your readers can get a precise visual. If they’re going through the chapters not fully picturing what’s happening, they won’t be fully invested.And readers who aren’t invested don’t become fans. And they don’t leave reviews. And they don’t buy any other books you publish.Strong verbs take a basic sentence and form a very specific image in the reader’s mind. Doing this throughout the entirety of your book will leave your readers feeling as if they just stepped out of an entirely different world.And that’s exactly what you want.Take these strong verb descriptions for example:She walked into the room, her cape trailing after her.She charged into the room, her cape billowing after her.She strutted into the room, her cape flowing after her.Each of these sentences is extremely simi lar in what they tell you; a girl with a cape entered a room.But changing the verbs from â€Å"walked† to â€Å"charged† to â€Å"strutted† alters the way in which she entered.It tells you the how.And knowing how an action takes place sets up far more than just the image for the reader. It tells them the mood the character’s in, increases suspense in some cases, and even creates anticipation for what’s to follow.#2 More impactful emotionsThe goal of your book (and any book, really) is to make your reader feel something. You want to stir emotions in them.That’s why they read books. That, and they want to be transported to a different world, which strong verbs are also used for.But one of the main reasons to use strong verbs in writing is to create a more emotional impact.When you want to create a strong reaction in your reader, no matter what type of reaction that is, you need to use strong verbs.Here’s an example of creating more anx iety or anticipation in your readers:My heart was beating so fast I could hear it.My heart crashed against my ribs, echoing in my head.Which sentence gives you a clearer picture of the anxiety that must be felt?The second, right.Because replacing â€Å"was beating fast† with â€Å"crashed against my ribs,† shows you just how hard my heartfelt. And thats the difference between a weak verb and a strong one.Strong verbs in writing are the KEY to showing and not telling in order to hook readers.Click To Tweet#3 Helps you show, not tellBy now you know just how important showing versus telling is in writing. And one powerful way to show more and tell less is to use strong verbs.It forces you to think more about the visual you’re trying to show the reader instead of just telling them what happened.Because showing creates a stronger emotional connection between the reader and your book, replacing weak verbs with more powerful ones will hook your readers.For those of yo u who struggle with showing and not telling, focusing on using better verbs will help tremendously.#4 They reduce weak adverbsWhen you’re writing, you may have a tendency to write sentences like, â€Å"I gripped the steering wheel firmly.† While this doesn’t look like a terrible sentence, it also doesn’t convey a very strong visual.Whenever you have an adverb, you should replace it with a stronger verb. That’s all an adverb is. It gives your weak verb a boost but it doesn’t actually make your sentence any stronger.Instead, replace â€Å"gripped firmly† with a powerful verb like â€Å"clenched† or â€Å"squeezed.†Ã¢â‚¬Å"I clenched the steering wheel† is a much stronger sentence that gives a clearer visual.Go through your writing and pick out some adverbs to replace. Your writing will be better because of it.#5 They make for more concise writingHave you ever picked up a book that looked decent enoughand even had an awesome title only to start reading and be turned off by how wordy and jumbled it is?Strong verbs prevent this.When you replace weak verbs and adverbs with a single stronger verb, you get rid of the excess writing that can make reading harder.It also saves you a ton of time cutting words during the editing phase.Because you’re using one word to create a strong visual, you won’t have to write more trying to describe how it looks to you. That strong verb does the job for you.This also allows for easier, more fluid writing and reading.Strong action verbs for better writingI won’t lie. One of the most frustrating things to read is a book that lacks strong verbs in scenes that are meant to be full of action.You’ll find this most often in fiction, but nonfiction books can be just as (if not more!) guilty of this. When you have a story that should leave the reader’s heart pounding but it doesn’t even raise their eyebrows, you have to do some diggi ng to improve.Heres a list of strong action verbs to improve your writing:What to do NextIt’s not enough to just know what strong verbs are and how to use them. You actually have to put these tips to use in your own book and implement what you’ve learned.Here’s what you can do right now to improve your writing and your book!#1 Attend your FREE trainingAll of this advice is relatively useless without a plan for finishing, marketing, and self-publishing your book.

Drive-By-Wire Cars are the Future essays

Drive-By-Wire Cars are the Future essays Concomitant cars have large pedals, steering wheels, and dashboards. Enter such a car and one will immediately become circumscribed. Ones feet must go on the pedals and ones hands must be placed on the steering wheel (Bertone). Likewise, true comfort can never be plenary until one is able to shift positions. Nevertheless, drive-by-wire technology is anon going to change all that by replacing clunky and inaccurate mechanical systems with highly advanced and precise electronic sensors (Brauer). Accordingly, car manufactures are developing state of the art drive-by-wire systems to ameliorate vehicle safety and comfort while simplifying car design and production. Despite this, there are huge logistical, cultural, and legal hurdles. Society will have to build smarter roads, perhaps including such features as fine-tuned global positioning system transmitters (Wylie). Moreover, people will have to give up control of their cars. Additionally, current drive-by-wire prototypes typically cost m illions and are not even legal to drive on most streets. Consequently, What well see more of in the next 10 years is driver-assistance features, such as lane keeping and spinout control (Wylie). Every year automobiles become just a little bit more refined as new technology replaces old ones. It started with luxuries like electric start and hydraulic brakes and continues today with direct injection, yaw control, and, of course, drive-by-wire technology (Brauer). In retrospect, the 1921 Duesenberg was the first production car to have hydraulic brakes, considered revolutionary then. Using hydraulic fluid pressurized by a master cylinder, the system exerted extreme pressure to activate and expand the drums in traditional drum brake systems and the piston calipers on modern disc brakes (Time Warner Cable). However, over 80 years later even with advent of anti-l...

5 Benefits of Accepting an Internship After College

5 Benefits of Accepting an Internship After College Not everyone can graduate college and go straight into a full-time well-paid position. And those who do are often at entry-level. Why not consider a post-graduate internship: one year of servitude in exchange for possible future advantages? Here are 5 benefits  of accepting an internship after college.1. Buy timeA year-long internship gives you an entire year of freedom to figure out where your interests and skills really lie and what sort of job you’d most like to be your first. It’s the best time to take stock and find the track that’s right for you, before plonking your train onto it and charging off into your professional life for good.Soul search, network, look for new opportunities, figure out in which direction you’d most like to push yourself. And remember that a year is short and the time will likely fly.2. Try on a new cityIf you land an internship in a new place, consider it an opportunity to try that city out before committing with a permanen t move. If towards the end of your internship, you find yourself less than thrilled with your physical surroundings, you’ll know where not to direct your job applications.3. Boost your resumeBottom line: internships make your resume look better. Most companies actually consider them a prerequisite for most quality jobs. Having this year on your resume can actually give you a competitive edge when you hit the job market for real.4. Get paidMost unpaid internships get away with being unpaid by offering college credit. But you’ve already graduated! They’ll have to pay you, however meekly. Take advantage of this by working extra hours whenever possible. You’ll prove your diligence and devotion, and start paying off those student loans!5. Use your degreeDon’t want to work alongside a sophomore who just wants to spend a semester in the city partying? Don’t worry. There are internships out there especially designed for college graduates. These will value your accomplishments and help you to make the most of your degree, rather than making you feel you’ve taken a step backward.In short, never underestimate the power of the internship to jumpstart your career.

IP 5A English Essay Example | Topics and Well Written Essays - 250 words

IP 5A English - Essay Example Despite this, I felt that this course gave me a direction that I needed. Previously, I had been unable to process as to how I should approach my passion for writing but this course opened a lot of opportunities for me. This did not come all of a sudden but with each assignment, I felt that I came closer to my goal. My biggest achievement in this course was the confidence I got. I learnt that writing is an acquired ability and practice will help me improve upon that ability. The discussion questions further allowed me to understand my strength in the class. My only disappointment is that I was not able to give my cent percent to this course. The stress from hospitalization prevented me from exploring my writing talent to the best of my abilities. I think I would have excelled if not for the deterrent. But despite this, I know that this course gave me the framework from which I could built my future in writing. I believe more reading on this course would have helped the students to explore more writing styles and perspectives. Overall, this course gave me a lot to move forward with. The discussion questions opened up not just new perspectives but better

Obama presidency and the media Essay Example | Topics and Well Written Essays - 2500 words

Obama presidency and the media - Essay Example In this respect, managing the president’s message is of paramount importance to the White House’s communications operations because it inevitably determines how the message is perceived both internally and externally, how it is interpreted, thereby influencing its outcomes respectively. This paper is an update chapter to Martha Joynt Kumar’s â€Å"Managing the president's message†, examining how the Obama administration has coped with new media and traditional media, and the effectiveness of the administration in conveying its message. Additionally, the paper comments on how the old and new media have covered the Obama presidency from the perspective of a news consumer on the premises of whether or not one can make judgment or take action based upon the information received by the news media. Out of seven US presidents in the period following World War II including Eisenhower, Nixon, Reagan, Clinton, and George W Bush developed effective communications ope rations because both the presidents and their staff understood the significance of explaining to the public the administration’s priorities and strategies that promoted their achievement (Kumar 1). Unlike these five heads of state, Republican George H W Bush and Democrat Jimmy Carter, the two chief executives who had minimal interest in presidential communications since they did not consider them as fundamental aspects of their presidency, had a lot of difficulty winning support of legislators and the public on many issues. The four basic functions of the presidential communications operations include advocating, explaining, defending, and coordinating on behalf of the chief of state; how effectively the communications operations carry out these functions depends on various factors. These include the nature of what the administration is trying to sell, the chief executive’s communication competence, organizational components of the communications operation; most import antly, the organization of the communications operations needs central control, an infrastructure that satisfies the ever-large news reporters need for news, and a team of communications staff that has vast knowledge of reporters’ routines. The single most important benchmark for effective communications operations is the nature of policies and effective performances of the administration that is trying to sell them since even if the communications operations is so good at what it does, it cannot function on a backdrop of weak policy or weak implementers. Like his predecessors, the US president Barrack Obama also regularly communicates with his fellow citizens to inform them of his administration’s plans, decisions, as well as stand points on both global and domestic policy issues as need may dictate, but through new media. Coping with new and traditional media The Obama presidency values the significance of an effective communications operations mechanism for the effe ctive delivery of his messages and, unlike all other presidents who have also longed to reach the people directly, bypassing the big national news agencies, the Obama presidency has taken this quest to extreme heights altogether with his thorough disdain of the media (Rubin). The Obama presidency has shunned mainstream media, and while he engages with both print and broadcast media every day, the chief of state together with his staff

Relationship between GDP and the quality of life in Brazil Research Paper

Relationship between GDP and the quality of life in Brazil - Research Paper Example 81). Scope of Research The companies that are able to take the proactive mindset should have the capability to rebuild several business strategies with the objective of shaping a better future to taste success. The aim of the thesis is to devise a win-win situation that benefits the communities and the corporate bottom lines in a similar fashion. Thesis Statement What are implications of corporate involvement in order to solve social problems? Literature Review According to Dominic Barton, the operating environment of business is experiencing radical transformation. The going concerns are getting influenced by demographical, technological and societal factors. It can be anticipated that the above mentioned trends will affect the operating environment, but it is surprising to witness the confluence of the trends. He states a wave of transformation on the way. Elizabeth Stevenson sought to examine the size or the scope of the changes. The time calls for examination of whether the move is towards venturing into new markets, or ways to survive efficiently. The operating environment will be influenced in a different fashion if the move is towards overcoming the scarcity of resources or interacting with the authorities in a different way. According to Patrick Viguerie, examination of the demand pools will reveal that the revenue leaders of today will not be in a sustainable position in the coming years as far as unit leadership in emerging markets is concerned. This means that the new world will witness new leaders. He stated that companies opt to view the short term while deriving strategies. Several insertion points were put forwarded by Sven Smith. The strategies should be informed by some upfront section along with some trends that will contribute to promotion of new businesses. The section can be undertaken on a monthly or annual basis. Research is required to determine the degree of the position of the resources against the trends as well as to make a second st rategy if the undertaken strategy sinks (Belgard & Rayner, 2004, p. 4). The forecasted trends are easy to be embedded into the strategic planning processes but in a tricky fashion. A long term trend embedded into the planning process can be a throwaway in the initial stages. The existing challenge is to engage line executives in order to recognize the trends, and if the trends are analyzed as forces of nature, it would be hard to recalculate again into the trajectory of the business keeping in mind the subsequent shifts. The tasks ahead of the global corporation are to contend with the amount of restructuring and shifting of portfolios that are anticipated to occur in the coming decade. The corporations need to analyze the nature of the shifts and build a conviction and find the right moves that will contribute to the creation of value. The implications of the trends will be profound. The coming years will witness multiple headquarters as the organizations will like to expand their operations. The pools of talent that has been created around the world now are sufficient to deal with the expansion plans. A competitive advantage lies in bringing the talent pools into the leadership arrangements and assimilates them into the borderless corporations. In order to unleash the talent, the corporation will have to reengineer the society and the economy and provide

Competitiveness in the International Electronics Industry Essay

Competitiveness in the International Electronics Industry - Essay Example The author Morri tells us that though the electronic industry is fairly small in the Dominican Republic by global standards, it employs however about 10,000 workers and contributes to about 5 percent of total Dominican exports (8). He further states that the electronic industry is by far the most important industry for a small economy (8). But in order to increase the competition in the electronic industry, the manufacturers in the electronic industry must learn to work together with the public sector to increase the competition position of the industry (Morri 9). The Dominican Republic has to meet the challenges of the public demands in order to ensure their competition in the electronics industry. The number of workers employed tells us how much is going into the export part of the trade. In order to accomplish competitiveness in the electronic industry, certain factors must be adhered to such as the "improvement in the quality and the cost of the electricity supply. Assistance with both local and international supply sourcing issues including possible common import purchasing and horizontal integration for inputs such as fabricated metals and moldings. In addition, the encouragement of domestic and foreign investment in critical local suppliers in such areas such as sheet metal fabrication, printed circuit boards, transformers, and general electronic component distribution (Morri 9)." In Latin America, there is extensive foreign investment in the sector by companies such as Verizon, Tricom, Centennial and Cable and Wireless and the cost of a T1 which is a high speed internet is the lowest priced in the whole of Latin America (Morri 16). On the other hand, "Guatemala, Honduras and Nicaragua have competitively low labor rates per hour, but they are not currently considered a competitive threat to the Dominican Republic because all three countries lack a secure environment (Morri 24)." In the above scenario, foreign investment in the Latin American companies makes it possible to guarantee low prices on certain commodities and labor being cheap makes it not a very feasible environment and so lacks the basis for a competitive environment. Another country that is ideal to examine would be the Chinese and Mexican market in relations to the US market. These two countries supply a lot of their goods to the US markets. According to Morri (9), China and Mexico supply a lot of their electronics to the US markets. In doing so, the global supply chain from the US to Asia threatens competition including low levels of linkages existing between the electronic manufacturers and the lack of industry expertise in the policy making and FDI promotion framework (Morri 10). The US markets are highly dependent on countries such as Mexico and China to provide a steady supply of electronic goods. Another important factor in increasing competition in the electronic industry is to : "create a one-stop-shop to streamline red tape. Increase image of electronics and the electrical industry locally. Also, employ or train electronic industry experts in policy and FDI promotion. In addition, build electronic industry support infrastructure for industry investors including offering investors full pre-, during- and post investment services and support. It would also help to decrease exit as well as entry barriers to increase investor confidence (Morri

Mental Health Laws in the movies PRimal Fear and Nuts Essay

Mental Health Laws in the movies PRimal Fear and Nuts - Essay Example In the case of â€Å"Nuts† the defendant is declared to be mentally insane but seeks to establish that she is mentally competent to face trial. While these two films have a different orientation to the subject of legal insanity, both of them deal with the courtroom proceedings surrounding the establishment of this legal defense. In both these films, the defendants admit to their criminal acts, but it is the defense that puts forth the insanity plea in â€Å"Primal Fear†, while it is the prosecution that advances the insanity defense in challenging the defendant’s competence to stand trial. The film â€Å"Nuts† revolves around a call girl, Claudia, who has killed a client and has been declared to be insane by the resident jail psychiatrist. She comes from a respectable, middle class white family and her mother, a timid woman together with her stepfather, are eager to ensure that the psychiatrist’s finding is substantiated, so that their daughter is declared not guilty and the publicity involved with a public trial can be avoided. As the film progresses, Claudia’s hostile attitude which even extends to clipping her attorney on the nose, coupled with the huge chip she carries on her shoulder only appear to corroborate this diagnosis and make it even more difficult for Claudia to prove that she is mentally competent to stand trial for her crime. As the film unfolds, some of the character’s motivation and reasons for her lifestyle and attitude become clearer, one of them being the revelation about her stepfather’s abuse in childhood and h er mother’s tacit knowledge of the situation, while failing to address it. The film ends with Claudia successfully establishing her competence to stand trial, since her hostility and violent attitude is partly explained by her background. The film â€Å"Primal Fear† presents a defendant who is a vulnerable looking altar boy, accused of

Competitiveness in the International Electronics Industry Essay

Competitiveness in the International Electronics Industry - Essay Example The author Morri tells us that though the electronic industry is fairly small in the Dominican Republic by global standards, it employs however about 10,000 workers and contributes to about 5 percent of total Dominican exports (8). He further states that the electronic industry is by far the most important industry for a small economy (8). But in order to increase the competition in the electronic industry, the manufacturers in the electronic industry must learn to work together with the public sector to increase the competition position of the industry (Morri 9). The Dominican Republic has to meet the challenges of the public demands in order to ensure their competition in the electronics industry. The number of workers employed tells us how much is going into the export part of the trade. In order to accomplish competitiveness in the electronic industry, certain factors must be adhered to such as the "improvement in the quality and the cost of the electricity supply. Assistance with both local and international supply sourcing issues including possible common import purchasing and horizontal integration for inputs such as fabricated metals and moldings. In addition, the encouragement of domestic and foreign investment in critical local suppliers in such areas such as sheet metal fabrication, printed circuit boards, transformers, and general electronic component distribution (Morri 9)." In Latin America, there is extensive foreign investment in the sector by companies such as Verizon, Tricom, Centennial and Cable and Wireless and the cost of a T1 which is a high speed internet is the lowest priced in the whole of Latin America (Morri 16). On the other hand, "Guatemala, Honduras and Nicaragua have competitively low labor rates per hour, but they are not currently considered a competitive threat to the Dominican Republic because all three countries lack a secure environment (Morri 24)." In the above scenario, foreign investment in the Latin American companies makes it possible to guarantee low prices on certain commodities and labor being cheap makes it not a very feasible environment and so lacks the basis for a competitive environment. Another country that is ideal to examine would be the Chinese and Mexican market in relations to the US market. These two countries supply a lot of their goods to the US markets. According to Morri (9), China and Mexico supply a lot of their electronics to the US markets. In doing so, the global supply chain from the US to Asia threatens competition including low levels of linkages existing between the electronic manufacturers and the lack of industry expertise in the policy making and FDI promotion framework (Morri 10). The US markets are highly dependent on countries such as Mexico and China to provide a steady supply of electronic goods. Another important factor in increasing competition in the electronic industry is to : "create a one-stop-shop to streamline red tape. Increase image of electronics and the electrical industry locally. Also, employ or train electronic industry experts in policy and FDI promotion. In addition, build electronic industry support infrastructure for industry investors including offering investors full pre-, during- and post investment services and support. It would also help to decrease exit as well as entry barriers to increase investor confidence (Morri

Overview on coral reef re-plantation in X-Sea port area Essay

Overview on coral reef re-plantation in X-Sea port area - Essay Example In many parts of the world, the coral population has undergone a massive reduction due to various human factors. The health of the coral reefs has been steadily declining over the past few decades. Environmental pollution and rampant destructive fishing practices damage the delicate corals. Nutrients seeping into the sea from agricultural areas cause the algae in the sea to increase in volume. This causes smothering of the corals and leads to decrease in size of coral reefs. Quite recently, the decrease in the coral populations has been attributed to a dozen or more deceases that attack corals. Many of these diseases were unknown until recently. Coral reefs have also been affected by bleaching. Bleaching is the discoloration or loss of symbiotic zooxanthellae. In 1979 and 1980, several incidents of coral bleaching occurred at reefs around Okinawa, Easter Island, northeast Australia, and the Caribbean Sea. Outbreaks of bleaching also occurred in 1982 and 1983, including reefs off east Africa, Indonesia, and the west coast of Central and South America, and from 1986 to 1988 in areas such as Taiwan, Hawaii, Fiji, Mayotte Island, and the entire length of the Great Barrier Reef. The most extensive bleaching episode ever documented occurred in 1998 and affected reefs in the Indian Ocean, Pacific Ocean, Red Sea, Persian Gulf, and Caribbean Sea. In some areas 100 percent of the corals were bleached and more than 70 percent of the corals died. (Smith 2003) Taking the rapidly declining coral population and the vulnerability of corals various measure are being taken in many countries to prevent further damage to the corals. Some of these steps include strict vigilance on fishing practices, artificial plantation of corals etc. Methods of coral community rehabilitation included coral transplantation and translocation, reattachment of coral fragments, providing artificial substrata, coral cultivation, prevention measures, and mitigation of damaged coral reefs. (Yeemin No

Health Services Organization Essay Example for Free

Health Services Organization Essay Identify and evaluate at least three forces that have affected the development of the health care system in the U. S. The U.S. healthcare system has continually evolved due to the success of innovative fore-thought. These innovations have been both strategic and tactical, influenced by all segments of the health care industry. There are many key strategic and tactical innovations, I will elaborate on three forces I feel affect the development of the healthcare system in the U.S.; they are: Increasing cost of healthcare, affect of economics on the healthcare system, and labor force trends in the U.S. as it relate to healthcare. * The steadily increasing cost of healthcare in the U.S. 1. The Unites States is known as the land of plenty, but if you are a native of this great nation you might feel quite differently about that statement. Being poor here has to be much like being poor anywhere else in the world, poor is poor; what might be different is opportunity. The opportunity that we are going to discuss is that of the pursuit of receiving the best healthcare regardless of race, creed, national origi n, or station in life (being with or without money). The cost of health care in the U.S. neared 2.6 trillion dollars in 2010, this is a staggering amount of money; but what make it more astonishing is that this amount is ten times higher than the 1980’s sum of 256 billion dollars that was generated by health care system (population bulletin, 2008). The rate of funds generated by health care has slowed down in recent years, but still is expected to grow faster than the national income. Our country need to address this growing burden and not just let politician say it’s a major policy priority, really make it a priority for the overall good of our nation. Furthermore, the United States has been in a recession for most of the past decade, resulting in a high rate of unemployment and lower incomes for many Americans. These conditions have put even more attention on health spending due to its affordability. Since 2002, employer- sponsored health care coverage for employee and family premiums increased by 97%, making the pursuit of health care a burden on the workers and employers. Medicare covers disabled people as well as the elderly; while Medicaid provides medical coverage for low-income families. Medicare enrollment has grown tremendously over the years due to the aging baby boomers and Medicaid due to the recession. This has added considerably to government spending, straining both federal and state budgets. Health spending accounted for 17.9% of the nation’s Gross Domestic Product (GDP) in 2010 (kaiserEDU.com, Health Policy from the Henry J. Kaiser Family Foundation, Explained). * Economic affect healthcare. 2. Economics and health care is very simple to explain, especially by someone who has seen first-hand families being mentally tormented, because they could not afford dental care for their children, the check-up not covered by the company’s insurance to enable your child to be eligible to play football, or not being able to afford the prescription medication. There are many reasons families go without medical care, and far too many times it’s just because the individual is living just above the poverty line; which make them ineligible for assistance. I personally feel the affect economics have on healthcare or the lack there of, can never be explained any clearer; unless you are experiencing it yourself. * Labor force trends in the U.S. 3. In the 1960’s the U.S. labor force grow at the rate of 1.7 percent annually and continue to grow into the 70’s as the baby boomer (those born between 1946-1964 generation continued to reach adulthood and enter the workforce. During this same period women began to enter the workforce as never seen before causing the labor force to grow at a rate of 2.6 percent annually. One might not see this as a health issue, but with women joining the workforce men began to feel the pressure, causing health complication. Women on the other hand who at one time lived the life of a care free house wife, but now working in corporate America; has started to feel stress now heart attacks are the number one killer of the American female species. I know there was nothing glamorous about being a slave, but slaves lived with just about no illnesses or life threatening diseases, know blacks have the highest numbers when it comes to diabetes, HIV, and high blood pressure than any other ethnic group (but we are free), freedom should make us take life more serious and take care of ourselves as well as our children. During the 1980’s and 90’s fewer people entered the workforce, therefore the labor force growth slowed down and ranged from 1.6 to 1.1 percent. Even though we had covered on set of baby boomers we were about to see another set reach adulthood and struggle for a place in the workplace (U.S. labor force trends), in-turn means no health insurance because they are no longer their parents dependent. Healthcare is becoming a great necessity due to infectious diseases, the year of the flower children seem to have spread more than just flowers. Health Insurance Companies are not seeing their usual capital gain at this point, consequently a change in the way benefits are paid across overall is about to change. At one time people thought as long as they paid their insurance premiums the benefits would be there when or if it was needed (we learned to read the fine print). * Speculate whether or not these forces will continue to affect the health care system in the U.S. over the next decade. (Include a force that was not mentioned that you believe will impact the health care system of the nation. Unfortunately we show no immediate signs that the cost of healthcare will decline in the Unites States anytime soon. With all the bickering about healthcare, there’s one fact everyone seems to agree on: American medicine cost too much, especially considering what we’re getting for the money. As expert look toward the future, they don’t see the cost dropping dramatically anytime soon. Curbing the rise in healthcare cost depend on our ability to gain control of the many forces causing them to climb, technology has helped other industries lower costs by eliminating waste and increasing efficiencies, but its done the opposite in healthcare (Lisa Zamosky, Los Angeles Times, September 12, 2012. Without any dispute I’m sure we can all agree the economy will always affect healthcare and the people who need it, which will always be everyone. We all hope the economy will make a rapid recovery, but that’s just not how things work; just as it took decades of bad decisions to get us where we are today it’s going to take time for our nation to make a meaningful recovery. Over the next 50 years, the labor force is projected to grow even more slowly (at about 0.6 percent per year) as baby boomers retire. What will this mean as far as health care is concern; well as we grow older our need for healthcare on a regular base becomes more essential than it was when we were in our prime (population bulletin). Baby boomers, reaching senior living status promotes a different type of demand on the medical system; a tremendous demand for those employed by senior living facilities, as well as those providing home health care. * Evaluate the importance of technology in the health care system. Technology is the driving force behind the universe, if you are not a believer just think about the devices you use to get through your day; hence technology. Medical technology is constantly presenting us with new devices that make our lives more fulfilling, we have laser surgery, hip replacement, artificial limb to replace worn out or damaged ones, cataract surgery that have advanced so rapidly it’s amazing, can you believe a person can have heart surgery and be walking around the next day? This is all because of technology, it’s amazing. This is something we cannot put a price on, what is being learned and the advances being made due to research technology is saving lives as well as making lives better each and every day. References: KaiserEDU.com, Health Policy, from the Henry J. Kaiser family foundation, Explained, Cost and Spending. http://www.kaiseredu.org/en/Top ics/Costs-and-Spending Population Bulletin, Vol. 63 No. 2, June 2008, U.S. Labor Forces Trends By: Marlene A. Lee and Mark Mather (Marlene A. Lee is a senior research associate and editor of the Population Bulletin at the Population Reference Bureau. She holds degrees in public policy analysis and development sociology/demography from the University of North Carolina At Chapel Hill and Cornell University. Mark Mather is associate vice president of Domestic Programs at the Population Reference Bureau, where he coordinates several projects that communicate population research to advocacy groups, educators, the media, and the public. He holds a doctorate and a master’s degree in sociology/demography from the University of Maryland. http://www.prb.org

Sucrose Synthase Key Enzyme In Sucrose Metabolism Biology Essay

Sucrose Synthase Key Enzyme In Sucrose Metabolism Biology Essay Sucrose synthase is a key enzyme in sucrose metabolism. Sucrose metabolism is required by the plant to form carbon required for various processes in the plant such as respiration, starch and cell wall formation. The enzyme is encoded by a small multigene family where most plants have at least two isoforms of the enzyme. The kinetics of sucrose synthase show that different Km values and ratios of sucrose breakdown exist for the enzyme. The methods of extracting, assaying and purifying the enzyme are shown in the enzyme characteristics. Factors such as pH, addition of different buffers, metal ions, fungal volatiles as well as environmental factors such as anoxia have all been shown to affect sucrose synthase activity. The enzymes protein sequences have been phylogenetically divided up into six main groups using clustalw. Sucrose synthase is normally present in the cytoplasm but the availability of sucrose in the chloroplast and its ability to use ADP as a substrate would indicate that the enzyme may be able to act in the chloroplast as well as the cytoplasm. Sucrose synthase is an important enzyme in sucrose metabolism in plants cells. (Persia et al., 2008) The main route of entry of carbon from sucrose is commonly known to be sucrose synthase. (Bieniawska et al., 2007) This carbon is used for respiration and in the synthesis of cell wall polymers and starch. (Persia et al., 2008) The main form of reduced carbon in plants is sucrose. It is used to support growth and synthesis of reserve materials e.g. starch in heterotrophic sink tissues. (Matic et al., 2004) The UDP-glucose supplied by sucrose synthase is used for cell wall biosynthesis while working with the cellulose synthase complex. (Baud, Vaultier and Rochat, 2004) In most fruit tissues, an increase in sucrose synthase activity is alongside with sucrose accumulation. This would suggest that sucrose synthase plays a physiologically important role. (Islam, Matsui and Yoshida, 1996) Carbohydrates are transported from photosynthetic source tissues to sink tissues in the form of sucrose . The consequent cleavage of sucrose in the sink tissues is the first step for its use in various metabolic pathways. The sugar is cleaved in vivo by either sucrose synthase (Sus) or by invertase. Invertase catalyses an irreversible reaction where sucrose is cleaved into glucose and fructose (Matic et al., 2004) while sucrose synthase catalyses the reversible conversion of sucrose and uridine-diphosphate (UDP) into uridine-diphosphoglucose and fructose. (Hirose, Scofield and Terao, 2008) (Hardin and Huber, 2004) These enzymes play a crucial role in plant growth and development. (Abid et al., 2009) Sucrose Synthase is cytosolic (Ã…  ebkovà ¡ et al., 1995) and has been characterized in many different plant species such as maize (Hardin and Huber, 2004), rice (Odegard, Liu and Lumen, 1996) and sugarcane (Schà ¤fer, Rohwer and Botha (2005)). Its activity has been studied in many plant organs such as roots, leaves and seeds. (Ã…  ebkovà ¡ et al., 1995) For trees, cellulose biosynthesis is a highly regulated process in which carbon is permanently placed in their primary and secondary cell walls. Sucrose is the main carbon source for cellulose synthesis. The stem is made up of extremely active sink cells which utilise sucrose for cellulose synthesis. Sucrose synthase is the main sucrolytic enzyme in these cells that catalyzes the reversible conversion of sucrose into fructose and UDP-glucose which is needed for cellulose biosynthesis. (Joshi, Bhandari and Ranjan, 2004) It also plays an important role providing adequate sugar supply during anoxic stress. It has been shown that during anoxic germination of rice, sucrose synthase activity was enhanced whereas the activity of invertase was depressed. This would indicate that sucrose synthase is the enzyme predominantly responsible for sucrose breakdown during anoxia. (Joshi, Bhandari and Ranjan, 2004) Fig 1: Diagram of the cleavage and synthesis reaction of sucrose synthase (Rà ¶mer et al., 2004) Different isoforms of the gene are present in most plants. In the case of maize, two non-allelic genes were discovered for sucrose synthase but more investigation lead to the discovery of a third. At least three genes for sucrose synthase have been discovered in rice where the genes show differences in expression between tissues. RSus1 is expressed in root phloem while RSus2 is expressed in leaf phloem. (Schà ¤fer, Rohwer and Botha, 2005) When examining the different isoforms at an amino acid level it is appears that there is less homology between different sucrose synthase genes in a species than when the gene is compared to its corresponding gene in another species. In the case of maize, there is 75% homology between the SS1 gene and SS2 gene of maize but there is 90% homology between rice RSus1 and maize SS2 genes. In sugarcane, the SS1 gene is 97% identical at the amino acid level to maize SS1 gene. (Lingle and Dyer, 2001) Nolte and Koch (1993) undertook a study to determine whether sucrose synthase was localized to certain part of the vascular strand. It is well known that sucrose synthase is present in vascular bundles for example in transgenic tobacco plants phloem specific expression of a maize sucrose synthase gene has been observed. Their study, using immunohistochemistry, found that sucrose synthase was restricted to the cytoplasm of companion cells of the phloem and did not appear to be present in other organelles of the plant. (Nolte and Koch, 1993) The molecular mass of sucrose synthase can be determined by gel filtration. Sucrose synthase elutes from the column with a Kav value of 0.17844 which when using a calibration curve correlates to a molecular mass of 362kDa. Using SDS-PAGE gradient gel the molecular mass of each subunit can be estimated at 92kDa. This can conclude that sucrose synthase is a tetrameric enzyme with a molecular mass of 360kDa and four identical subunits of 90kDa. (Hardin and Huber, 2004) (Elling and Kula, 1993) It can associate with membranes and the actin cytoskeleton where its activity is known to be involved with cellulose synthesis. It does this by channelling uridine-diphosglucose to the growing glucan chain by the enzyme cellulose synthase. (Hardin and Huber, 2004) Analysis of Sucrose Synthase Gene Family: From the results of species examined to date, it is shown that sucrose synthase is encoded by a small multigene family. (Bieniawska et al, 2007) Most species of plants have at least two isoforms of sucrose synthase. These isoforms usually have comparable biochemical properties and highly homologous amino acid sequences. (Wen et al., 2010) Further analysis of transgenic and mutant crop plants show certain isoforms of sucrose synthase have specific functions in the plant. The rug4 mutation of pea removes the SUS1 isoform but has no effect on SUS2 or SUS3. This would indicate that these two isoforms are not able to make up for the loss of SUS1 in the seed or root nodule. It is clear that the loss of different isoforms affect the plant in certain ways. Loss of the SH1 isoform in maize has different outcomes from the loss of SUS1 isoform. SH1 is required for normal cell wall formation during endosperm development while both isoforms are needed for wild-type rates of starch synthesis. Why different isoforms have different functions is unclear. The same functions can be carried out in the cell by different isoforms but can occur in distinct cell types, developmental periods or environmental conditions. It is likely that different isoforms could have non-overlapping, particular functions in the same cell. (Bieniawska et al., 2007) It is difficult to decide on the precise roles of the genes in sucrose synthase gene family when there is not enough information in existence. Although there is some information available on some of the isoforms and theyre functions in the plant, no analysis of the functions of the gene family has been carried out. The model plant Arabidopsis is ideal for carrying out such an analysis. Six sucrose synthase genes are in the Arabidopsis genome. Based on comparisons of the amino acid sequences the isoforms they encode can be divided into three distinct pair groups. The isoforms SUS1 and SUS4 are 89% identical to each other but have less than 68% similar amino acid sequences to other isoforms. Similarly, SUS2 and SUS3 are 74% identical to other isoforms and are 67% less identical to the other forms of enzyme. SUS5 and SUS6 are 585 identical to each other but have less 48% similarity to the other isoforms. When examining other dicotyledonous species it appears that at least two of the thr ee pairs of isoforms are present. When phylogenetic analysis was carried out, it showed that the isoforms AtSUS1 and AtSUS4 are related to pairs of isoforms from pea (Fabacae), carrot (Umbelliferae) and potato (Solanacae). A pair of isoforms from Craterostigma plantagineum (Scrophulariacae) is closely related to the pair of isoforms AtSUS2 and AtSUS3 in the Arabidopsis. The pair AtSUS5 and AtSUS6 is related strongly to a pair of genes from rice. This evidence shows that it is unlikely that the three pairs of isoforms in Arabidopsis are as a result of gene duplication events. It is possible that each isoform has an exact function preserved in a wide range of plants. The members of Arabidopsis gene family are strongly differentially expressed in different organs of the plant through its development and in response to external stimuli e.g. environmental stress. This is seen in gene families of other plants studied. (Bieniawska et al., 2007) Fruit quality is determined by the type and quality of sugars present. A study of the sucrose synthase-encoding gene from the muskmelon fruit was carried out to evaluate how to genetically improve the quality of the fruit. This is done by finding the sugar components in fruit, to identify the enzymes involved in sugar metabolism and distinguish the relationship between sugar accumulation and the activities of related enzymes. It is thought that sucrose synthase is the enzyme involved in metabolising sucrose in developing muskmelon fruit. To examine this, a full length cDNA strand encoding sucrose synthase was extracted from a muskmelon fruit by RT-PCR and RACE and identified as CmSS1. Real time PCR analysis showed that CmSS1 expression changed in among different tissues of the plant e.g. root, stem, leaf. It showed that the mRNA levels are highest in the root and lowest in mature fruit. Fig 2: The patterns of CmSS1 transcript abundance in the different tissues of the muskmelon plant. These results were found using quantitative real-time PCR analysis of total RNA prepared from the root, stem, leaf, flower and mature fruit of muskmelon. During fruit development and ripening it was shown that CmSS1 mRNA was at its maximum level at five days after pollination and decreased steadily during fruit development until it reached its minimum level of maturity. This was discovered using again real-time RT-PCR analysis of mesocarp tissues from five days of pollination to ripening. Fig 3: This graph depicts the patterns of CmSS1 transcript abundance in developing muskmelon fruits found by using quantitative real-time PCR analysis of total RNA prepared from muskmelon. (Wen et al., 2010) The sugar content and SS activity were analysed to show the functions of CmSS1 in regulating fruit quality. It showed that very low concentrations of sucrose are present in young and unripe muskmelons. Between 20 and 30 days after pollination there is a massive rise in the amount of sucrose in the fruit. Sucrose synthase activity increased in the direction of sucrose synthesis and decreased in the direction of sucrose cleavage through fruit development. (Wen et al., 2010) Fig 4: The depiction of sucrose content and sucrose synthase activity during muskmelon fruit development. The first chart shows sucrose content during fruit development. The second shows sucrose activity in the sucrose synthesis direction and the third shows sucrose cleavage direction during muskmelon fruit development. (Wen et al., 2010) Enzyme Kinetics of Sucrose Synthase: An investigation was carried out by Schà ¶fer et al. to the find the properties of three sucrose synthase isoforms present in sugarcane. Kinetic analysis indicated that the three sucrose synthase genes in sugarcane are different isoforms, with major differences in Km values and the ratios of sucrose breakdown synthesis. The kinetic characteristics of the SuSyA and SuSyB isoforms, both expressed in the leaf roll, differ greatly. It was found that SuSyA has almost three times higher affinity for sucrose than the SuSyB isoform whereas SuSyB has a much greater affinity for UDP than SuSyA. Based on the differences in their kinetic properties it can be concluded that SuSyB and SuSyC are different isoforms of sucrose synthase. SuSyC has roughly ten times higher affinity for UDP compared to the other two isoforms. (Schà ¤fer et al., 2005) Fig 5: The graph shows the Lineweaver-Burk plot of 1/v against 1/S for the isoforms SuSys A, B and c where UDP was the variable substrate. The concentration of sucrose was kept constant at 320nM. The Km values were determined from the non-linear fit of the data to the Michaelis-Menten equation. (Schà ¤fer et al., 2005) When examining sucrose synthase in soybean nodules Morell and Copeland (1985) found the kinetic constants of UDP, UDPglucose, sucrose and fructose by fitting the data to the following two equations: 1. v = VA/KiaKh + KhA + KhB + AB 2. v = VA/Ka + A + A/Ki The kinetic constants for ADP, CDP and ADPglucose were found using non linear regression analysis of initial velocity data. Fig 6: Graph showing the effect of sucrose concentration on the cleavage activity of sucrose synthase in soybean nodule. The lines show the fit of data to equation 1. The reaction mixture were composed of 20 µmol Hepes-KOH buffer (pH 7.5) 2 µmol UDP, 1.5 µmol NAD, 25 µg UDPglucose dehydrogenase. Each symbol represents a different concentration of sucrose. The dark circle shows 3.2 µM, the clear circle shows 4 µM, the dark triangle shows 6.25 µM, 10 µM is shown by the clear triangle and the dark square depicts 20 µM. In the cleavage and synthesis direction standard Michaelis-Menten kinetics are observed. The variation of concentration of sucrose at different concentrations of UDP gave an intersecting pattern of linear double reciprocal plots. (Morrell and Copeland, 1985) Parameter Value V (U/mg protein) 13.3 ±2.0 Km sucrose (mM) 31.3 ±7.1 Ki sucrose (mM) 31.9 ±13.1 Km UDP (mM) 0.005 ±0.002 Ki UDP (mM) 0.005 ±0.001 Fig 7: Table showing the kinetic parameters for the cleavage reaction of sucrose synthase in soybean nodules. (Morrell and Copeland, 1985) Fig 8: The graph depicting the effect of UDPglucose concentration on the synthesis reaction of sucrose synthase activity in soybean nodules. The reaction mixtures contained 20 µmol Hepes-KOH buffer, 15  µmol fructose, 5 µmol MgCl2, 0.4  µmol P-enolpyruvate, 0.15  µmol NADH, 20 µmol KCl, 25 µg pyruvate kinase 25 µg lactate dehydrogenase and the required amount of enzyme. As in the previous graph, the amount of UDPglucose was varied in the presence of 2.5mM (dark circle), 3.2mM (clear circle), 4mM (dark triangle), 5mM (clear triangle) and 8mM (dark square) fructose. The results on the graph are representing the fit of data to equation 1. When the concentration of UDPglucose was varied at the concentrations of fructose in the graph, an intersecting pattern of linear double reciprocal plots was seen. From fitting the data from the graph to equation 1, it is noted that substrate inhibition would have occurred at a concentration greater than 15mM fructose. Parameter Value V (U/mg protein) 14.3 ±1.2 Km fructose (mM) 3.7 ±0.8 Ki fructose (mM) 19.6 ±9.9 Km UDPglucose (mM) 0.012 ±0.006 Ki UDPglucose (mM) 0.064 ±0.014 Fig 9: table showing the kinetic results by fitting the figures from the graph to equation 1. When partially purified SuSyA, SuSyB and SuSyC were blotted to a nitrocellulose filter the results showed that all three isoforms are approximately 94kDa. (Schà ¤fer et al., 2005) The would correlate to the findings of Hardin et al and Lothar et al who stated that sucrose synthase is tetrameric enzyme made up of four 90kDa subunits. Fig 10: Immunoblot of sugarcane SuSy. A crude extract of protein from leaf roll was loaded into lane 2 while partially purified isoforms of SuSyA, SuSyB and SuSyC were loaded to lane 3, 4 and 5. The molecular weight ladder was used to identify the bands see in each lane. (Schà ¤fer et al., 2005) Characteristics of Sucrose Synthase: Extraction of Protein: The method for extracting protein from the leaves of maize (Zea mays), rice (Oryza sativa) and tobacco was done as follows: 1-3g of leaves was ground in liquid nitrogen and the powder was mixed in the ratio 1:2 with extraction buffer. The buffer was made up of 0.1M tris-HCl, pH 8, 10mM DTT and 1% polyvinylpolypyrrolidone. The samples were then incubated on ice for 15 minutes and then centrifuged at 1,000g for 10 minutes at 4oC. The pellet was then removed and the supernatant was re centrifuged at 100,000g for one hour at 4oC. After this final centrifugation, the pellet and supernatant which contained the soluble proteins was resuspended in sample buffer for electrophoresis. (Persia et al., 2008) When extracting protein from rice seeds, a similar procedure is followed. Seeds weighing roughly 50-100mg at various stages of growth were homogenized in 400 µl of extraction buffer and kept at 4oC. The buffer was made up of 50mM Tris-HCl, pH7.5, 1.0mM DTT, 1.0mM EDTA and 2mM PMSF. Ammonium sulphate fractions (30-50% w/v) were precipitated and then resuspended in dialysis buffer made up of 50mM Tris-HCl, pH 8.0, 5mM MgSO4, 5mM 2-mercaptoethanol. This was then dialyzed overnight at 4oC. (Odegard, Liu and De Lumen., 1996) The method for extracting protein from tobacco pollen tubes is slightly different to those mentioned previously. The pollen first was slowly thawed from storage at -20oC and hydrated in a humid chamber overnight. It was then germinated in BK medium and allowed to germinate at 25oC for three hours. After this period had elapsed, the pollen was collected by centrifugation at 1,000g for 5 minutes at 25oC. It was then washed twice with BRB25 buffer which is made up of 25mM HEPES, pH 7.5, 2mM EGTA and 2mM MgCl2 and 15% Suc. After washing, the pollen was resuspended in lysis buffer and lysed on ice using a motor-driven Potter-Elvehk-jem homogenizer. The lysis buffer used was made up of BRB25 buffer along with 2mM dithiothreitol, 1mM phenylmethylsulfonyl fluo ride (PMSF), 10 µL/mL protease inhibitors, 1mM NaN3 and 10% mannitol. After lysis was carried out, the samples were centrifuged at 1,000g for 10 minutes at 4oC. The supernatant was centrifuged again at 4oC for 45 minutes at 100,000g over a 20% (w/v) Suc cushion. The supernatant was then collected as it contained the soluble protein fraction. (Persia et al., 2008) Enzyme Assays: After extracting protein, the sucrose synthase activity in sugarbeets was found using a spectrophotometric end point assay. The activity of the enzyme was monitored as fructose formed at 35oC. This was carried out in a solution that contained 250mM sucrose, 2mM UDP and 100mM MES. The control was carried out by assaying for activity in the absence of UDP. The total protein concentration was determined using the Bradford method where bovine serum albumin was the standard. (Klotz and Haagenson., 2008) When assaying for protein from rice, the Bradford method was followed to determine protein concentration as was done in Klotz et al. 40mg of protein was used per assay. The assay was carried out in 20mM MES pH 6.4, 200mM sucrose and 4mM UDP for 15 minutes at 30oC. The reaction was stopped by boiling for 2 minutes and the fructose levels were measured. The control tubes did not contain UDP. (Odegard, Liu and De Lumen., 1996) When examining the effect of sucrose synthase on carbon partitioni ng a similar method was followed for assaying the protein. Sucrose synthase was assayed in the direction of sucrose breakdown using 50 µl poplar plant extract. The tetrazolium blue assay was followed to determine the amount of free fructose. As in previously mentioned assays, the absence of UDP in the assay acted as a control. The total protein content was found by employing the Bradford (Bio-Rad) protein assay. (Coleman, Yan and Mansfield., 2009) A similar method was followed for carrying out an assay for the enzyme on tomato tissue. The reaction mixtures contained 50mM Hepes-NaOH buffer, 15mM MgCl2, 25mM fructose and 25mM UDP glucose. This was incubated at 37oC for 30 minutes and was terminated with the addition of 70 µl of 30% KOH. The enzyme blanks were terminated with the addition of KOH at 0 minutes. The tubes were then kept at 100oC for 10 minutes to destroy any fructose. The soluble protein content was determined using the Lowry method whereby bovine serum albumin was th e standard. (Islam, Matsui and Yoshida., 1996) Alkaline copper solution is added to each tube and allowed to stand at room temperature for roughly 30 minutes. Dilute folate reagent is then added to each tube rapidly and after 30 minutes the absorbance is read at 750nm. (Lowry et al., 1951) The results were measured as  µmole of sucrose per minute per mg protein. (Islam et al., 1996) When assaying for sucrose synthase in the cleavage direction Rà ¶mer et al used recombinant SuSy1 gene from potato. In a volume of 100 µl HEPES buffer with a concentration of 200mM and pH 7.6 recombinant sucrose synthase was incubated along with 2mM UDP and 500mM sucrose for ten minutes at 30oC. HPLC analysis was used for the formulation of UDP-glucose. The Bradford assay was used to determine protein concentrations as was carried out by Klotz et al and Coleman et al. The activity of the enzyme was also tested with the nucleoside diphosphates dTDP, CDP, ADP and GDP at 2mM. For assaying recombinant e nzyme in the synthesis direction a similar method was followed as when assaying for standard enzyme. Recombinant sucrose synthase was incubated in a total volume of 100 µl HEPES buffer where this time the pH was 8.0 and the concentration was as in cleavage direction of 200mM. 1mM UDP-Glc and 20mM D-fructose was also added to the mixture and it was incubated for five minutes at 30oC. The reaction was heated to 95oC for five minutes and HPLC analysis was used to establish the formation of UDP. The sucrose synthase activity was also tested using dTDP-Glc, CDP-Glc and ADP-Glc. (Rà ¶mer et al., 2004) Purification of Protein: After extraction of the protein from the crude extract, purification can be carried out. This can be done in a number of ways such as Batch adsorption with Sephadex A50, Anion exchange chromatography and Gelfiltration. SDS-PAGE can be carried out after purification to check the purity of the protein sample. The Sephadex A50 gel is loaded into a glass funnel and washed twice with deionised water. The gel was then washed twice with 300ml standard buffer. The protein sample was loaded to the gel and slowly sucked through the gel for 30 minutes. The gel bed was then washed with 300ml standard buffer and then with 300ml standard buffer containing 100mM KCl. The last washing step contained 300mM KCl. 200ml of the first salt preparation was concentrated to 40-50ml by using a cross-flow ultrafiltration module with YM 30 ultrafiltration membrane that had been pretreated with 55 PEG 4000 solution. This was done to prevent the enzyme sticking to the membrane. In anion exchange chromatography a Sepharose Q column was first equilibrated with 300ml Hepes buffer. This was made up of 200mM pH 8 with 50mM KCl. 70-80mg of protein sample was loaded and the elution was started using two different salt gradients. To prevent enzyme inactivation after elution all the fractions were titrated back to pH 7.2. All fractions that contained enzyme activity were pooled and concentrated by using ultrafiltration. Gelfiltration experiments are carried out on a prepacked HiLoad 16/60 Superdex 200 prep grade column that was connected to FPLC equipment. Four samples containing 2mg of protein were loaded and eluted with a flow rate of 1 ml min-1. The fractions were then pooled and stored at -20oC in 500 µl aliquots. (Elling and Kula., 1993) To determine the purity of the protein, SDS-PAGE is carried out. This is done by loading 100 µg of protein samples to a 125 SDS-polyacrylamide slab gel that was overlaid with stacking gel. The electrophoresis was carried out at 4oC and at 40V for 16 hours a nd followed by 200V for one hour. Coomassie blue R 250 was used to stain the gel followed by destaining. (Kumutha et al., 2008) Factors that affect Sucrose Synthase Activity: Ã…  ebkovà ¡ et al (1995) stated that sucrose synthase has two different pHs for optimal activity. In the cleavage direction it was found that most enzyme activity was observed between pH 6.0 and 8.5 at temperatures between 50 to 55oC. In the synthesis direction, a pH between 8.5 to 9.5 and a temperature of 35oC was optimal for enzyme activity. (Ã…  ebkovà ¡ et al., 1995) This would correlate with the findings of Morell and Copeland (1985) who found that optimal activity of the enzyme in soybean was at pH 6 in the cleavage direction and at a pH of 9.5, sucrose synthase activity in the synthesis direction was at its highest. It was also found that at a pH of 7.5 the cleavage and synthesis activities were their highest. (Morrell and Copeland., 1985) Elling and Kula (1995) examined the effect of buffers TES-NaOH, MOPS-NaOH, TEA-NaOH and Tris-HCl on the pH optimum of sucrose synthase activity. These were determined using UDP and TDP as substrates for the reaction. They found that the e nzyme had its highest activity in Hepes-NaOH buffer. When MOPS-NaOH and TES-NaOH buffer was used, only 60-80% activity was noted. (Elling and Kula 1995) It was also found that the velocity of the reaction could be increased by increasing the temperature where optimal activity was seen between 50 and 60oC. Xu at al (1989) reported that potato and bean are also able to withstand these high temperatures. However once the temperature goes above 60oC enzyme activity starts to decreased rapidly and was destroyed once the temperature reached 70oC. (Xu et al., 1989) The cleavage of sucrose by the sucrose synthase enzyme was investigated to find the rate of cleavage reaction using different nucleosidediphosphates as cosubstrates. They found that the rate of reaction was UDP>TDP>ADP>CDP>GDP. Echt and Chourey (1985) found similar results when examining nucleotide specificity. They found that substrate specificity for SS1 and SS2 were UDP>TDP>ADP>CDP>UTP where each substrate was at a concentrat ion of 4mM. (Echt and Chourey 1985) Low levels of heavy metal ions such as mercurate inhibited cleavage activity of the enzyme. This would lead to the assumption that sulfhydryl groups are involved in the catalytic process. It is also inhibited by Tris-HCl and by small concentrations of MgCl2 and MnCl2. (Ã…  ebkovà ¡ et al., 1995) Cations were shown by Elling and Kula (1995) to have a slight influence on enzyme activity. The activity was lessened slightly (10%) by the presence of 1mM Mn2+ and Mg2+ ions with UDP. The enzyme is completely inactivated in the presence of 1mM Cu2+ or Fe2+. (Elling and Kula., 1993) A recent study was undertaken to examine the effects of volatile emissions on carbohydrate metabolism. Studies on this area have taken place before but it is usually examining the results of physical contact between the host plant and the microbe. No work has taken place until now on the effect on the plant in the absence of physical contact. Many microbes such as Pseudomonas spp, Strepomyces spp, Penicillin spp and a selection of truffles produce ethylene. This gaseous plant hormone plays an important role in many aspects of plant growth and development such as seed germination, root hair initiation, fruit ripening and starch accumulation. In the work of Ezquer et al (2010), the possible effects of volatiles released from gram-negative bacteria, gram-positive bacteria and fungi on starch metabolism was studied. The results showed that the volatile compounds released by microbes promoted high levels of starch accumulation in mono- and dicotyledonous plants. It also revealed fungal vo latiles (FVs) promoted massive changes in expression of genes involved in many important processes in plant such as metabolism of carbohydrates, amino acids, sulphur and lipids, energy production, protein translation and stability, cell wall biosynthesis and photosynthesis. However no changes were noted in the expression in some of the genes that coded for proteins involved in starch and sucrose metabolism such as plastidial hexokinase, plastidial phosphoglucose isomerase, plastidial adenylate kinase, alkaline invertase and UDPglucose (UDPG) pyrophosphorylase. It was found in the study that FVs strongly upregulate the expression of Sucrose Synthase in potato leaves. The plants were cultured in the presence and absence of FVs emitted by A. Alternata. This caused a massive enhancement of expression of Sus4 isoform. A 29.4- and 31.63-fold increase was observed in expression when the plants were cultured in the presence and absence of sucrose. This isoform of the enzyme controls the accumulation of ADPG, UDPG and starch in potato source leaves and tubers. Analyses of the intracellular amounts of starch and nucleotide-sugars in the leaves of the plant show a positive correlation between patterns of enzyme activity and starch, UDPG and ADPG amounts. This was noted when the leaves were cultured in the presence and absence of FVs. Western blot analyses and quantitative RT-PCR confirmed also the increase in expression. (Xu et al., 1989) Environmental Factors affecting Sucrose Synthase Activity: Anoxia: Waterlogging is where oxygen supply is blocked to root leading a severe decrease in the amount of oxygen available to the plant. This leads to inhibition of root respiration that causes a major decline in energy of root cells affecting vital metabolic processes of the plant. This is restriction of oxygen supply is known as anoxia. The presence of glucose in an anoxic incubation medium drastically decreases meristem death and studies have shown that sucrose synthase is the enzyme mainly responsible for sucrose breakdown under anoxia. (Kumutha et al., 2008) The increase in glycolytic demands caused by these demands is the cause of increased sucrose synthase expression. This has been demonstrated in many plant species e.g. sucrose synthase gene is induced in wheat and in rice when oxygen levels are low. (Ricard et al., 1998) Harada et al (2005) also found an increase in sucrose synthase activity in pondweed turins while under anoxia. (Harada et al., 2005) Klotz and Haagenson (2008) foun d that sugarbeet contained two genes for sucrose synthase activity-SBSS1 and SBSS2. They demonstrated that anaerobic conditions caused a large increase in the transcription levels of SBSS1 and a quick increase and succeeding decline in SBSS2 transcription levels. However this did not correlate with a significant increase in sucrose synthase enzyme activity. A 23% increase in sucrose synthase activity was noted after initiation of anaerobic conditions but otherwise the activity of the enzyme did not differ greatly to that of the controls. (Klotz and Haagenson., 2008) Fig 11: The graph outlines the different rates of sucrose synthase activity in the con