The Business Plan Of 643744 Company

The Business Plan Of 643744 Company This business plan provides to shareholders and customers an analysis and evaluation of the prospective marketing capability, product capability, and profitability of small computer industry-643744 Company in three years by running business simulation SimVenture. 643744 Company designs products, and sales superior computers, focusing on the professional services customers. Based on the investigation of resources, strengths, and capabilities competitive, customer, and environmental issues, company establishes their vision, mission, and achieving goals over three financial years. The companys vision is Walking in the forefront of times. The mission and goals have been represented in table 1. Currently, the basic salary of the owner is planned  £1500 per month. Table : The Companys mission and goals in following 3 years Year Mission Goals 1st Year Our products must be: Sophisticated in quality Offer good value Beautiful artistic style, and Philosophically inspiring Our role is to play a vigorous part in markets. We can improve people lives to be happier and effective through their use of our products. Devising the idea design to meet customers needs Accomplishing above 50 orders 300 products 2nd year To be the No.1 computer service company in the world and among the premier industrial concerns in terms of quality, profitability and growth, through innovative leadership. Enlarge its markets Accomplishing 100 orders 1000 products 3rdYear To be the preeminent building block supplier to the worldwide electronics business. Achieving 150 orders monthly 1500 products Targeting global markets 643744 Company followed the strategic planning process (Ferrell Hartline, 2011:32), which was separated into 4 sections. It focuses on integration of effort to achieving operation, sales and marketing, human resource (HR), and finance stated objective (Ferrell Hartline, 2011:39). In the sales and marketing section, it analyses the market prospect though marketing research and customer research. Then it provides marketing strategy and risk management plan, dealing with marketing promotion via advertising channels to achieve customer needs and build customer values. In operation, it involves in the strategy of the Supply chain management (SCM), the Just-in-time (JIT) inventory control, and outsourcing management. In HRM, it focuses on employee recruitment, training and evaluation. Functional strategy aims to meet companys mission, goals and objectives. It also evaluates the effort on organizations sales, costs, image, and profitability. Finally, the business plan explains financial r isks management, and reveals the cashflow and the profit and loss forecasts over three years. This program conducted has limitations on that the business will not be registered for VAT in software SimVenture calculation. Sales marketing Marketing research Via a range of Ad hoc research (Jobber Fahy, 2009:85), company targeted at professional services market. According to the market report (Figure 12 in appendix), by contrast with other 5 segments, the professional services have 2415 market shares with 24 months order frequency. It has to succeed by good quality, good style, and above javerage features. Thus, the fixed cost can be generally calculated in  £450 per unit. It has more potential interests under the typical price than solo business and service companies. Competitor research 643744 Companys competitive strategy focuses on new product developing strategy to be a low-cost manufacturer and improve the cost-effectiveness of production process (Czinkota, 2000:268-269). According to the competition research and analysis (Figure 13 and table 19 in appendix), companys competitor, Parry systems, is a medium size maker of fashionable computers in professional services market. In the company competitive strategy, firstly, managers can influences on consumer behavior in buying situation by affording information-rich communication through advertising and the sales forces (Fill McKee, 2012:75-79). Thus, company supports marketing promotions applying institutional and product advertising (Ferrell Hartline, 2011:299), via different channels. Additionally, though the Voice of customer (VOC), company can offer marketing programs to raise customer satisfaction and create long-term reputation (Ferrell Hartline, 2011:354). Czinkota (2000:104) commented that a successful c ompany should focus on the differentiation of chief players in the marketplace, which will react to the companys changes in business tactic, rather than customers reactions on new product or business strategy. Sales and marketing strategy On the marketing promotion strategy, The 643744 Company performed in transation market, personal selling, and distributor sales channels, taking suggestion from Jobber and Fahys theory (2009:230). It explains that the consumer and trade promotion could be considered through those 11 approaches (Figure 1), including price discounts, premiums, money off, allowances, free goods, and loyalty cards etc. Figure : Consumer and Trade promotion (Sources: Jobber Fahy, 2009:230) In the pricing strategy, the price changes and setting might occur over the products life cycle (PLC) (Ferrell Hartline, 2011: 239), which is also influenced by costs, competitions and marketing (Hobber Fahy, 2009:194). Homozi (et al., 2002:755) believes the most central point is where the pricing strategy realistically places the business in comparison to the competition. Based on the monthly customer research and break-even analysis, company apply the price penetration strategy (Czinkota 2000:513), and pricing promotion strategies. However, the strategy is a continuous process rather than a single event (Pollard and Hotho, 2006). The further information is illustrated in the table 2: Table : marketing promotion planned strategy The 643744 Company focuses on the international market in the future, which is the most effective management of B2B operations (Lages, Abrantes, Lages, 2008, cited in Sharland, 2001; Katsikeas, 2006). However, this is remarkable considering that although small business is not able to compete on price with large international companies (Lages, Abrantes Lages, 2008). Day (1984:102-103) recommends, management general strategies could be considered by increasing sales volume, or improved productivity or margins (Figure 2). Nevertheless, it is difficult to evaluate the factual scanty order and insufficient profitability in marketing strategy. Customer satisfaction depends on their exceed expectation, which is created by pre-buying experiences and discussion with other customers (Jobber Fahy, 2009:6). Therefore, strategically, the product design is the key to entry into professional service market, which can be succeeded in minimum two months. Then managers need to improve their product design by monthly customer research, due to the PLC theory (Fill McKee, 2012:44). Additionally, Company determines to contract out automatically by normal quality within 5 minutes on testing to prevent faulty products and build business reputation. Purchasing A successful supply chain management (SCM) can fetch competitive advantage and better margins (Fill McKee, 2012:182). Company selects ProSupply supplier with more flexible payment terms, which has been considered its insufficient order quantity at introduction stage of the PLC. As the marketing expanded, suppliers could be changed to Sourceline or MegaParts with more discounts (Planed in the table 4). One of SCM risk is the approach of cooperation with upstream associates to obtain efficient and effective supply of materials ¼Ã‹â€ Guido et al., 2008 ¼Ã¢â‚¬ °. Nevertheless, Fill and McKee (2012:182) believe that a penny saved here becomes millions saved there. The company relocates working place started into a Small Business Centre that provides partial units, beneficially for new business, and cheaper than other commercial premises. Due to the limitation of Small Business Centre, in the latter two years, managers will rearrange the office into a retail unit. The table 5 represents the detail with calculation. The started-up office resources (Table 6) are one standard office equipment and two premium tools. Having a car can be considered in the third year. In the later two years, the double quantities in each will be required at the end. People The company selected the hard and soft model of human resource management (HRM) that integrates hard and soft HR performance. Price (2011:548, cited in Storey, 1989) believes that People are led rather than managedà ¢Ã¢â€š ¬Ã‚ ¦ à ¢Ã¢â€š ¬Ã‚ ¦involved in determining and realizing strategic objectives Basic on the initial information of staff skills, business owner required more capabilities training on research, marketing, and sales, etc. it will be improved efficiently by training centre course. The working time will be under 60 hours per week. On the other hand, Setting-up a limited company in law is essential before the business owner goes full-time, which intends to implement after the first year. For the future business expansion, employment contract is required and employees are under controlled in 5 members. Creating statutory contacts with customers is obligatory at the beginning. Finance Forecasting Amir (et al, 2002) argues the lack of liquidity is the greatest threats to start-up business. Cashflow can help businessmen ensure when extra cash requires and plan accordingly. The company has capital  £10,000 in bank balance for starting business. The three-year cashflow forecast and the Profit and loss forecast have described in appendix 2, in the which, the predictive profit is  £311,182 over three years. Additionally, it is essential to prepare a contingency plan (presented in table 7). Through the process of contingency plan, dealing with the immediate consequences of the potential severity is to compensate for the changes, and shift to the minimized impact on the businesses viability. Table : Contingency plan of financial risks (Source: Author) Potential severity Solution Sales are less than forecasted Increasing marketing promotion To do more advertise or personal sales Reduce price Start-up finance takes three months longer to obtain Increasing working hours Using professional Consultant Liabilities, financial crisis in the short term Borrow money from the bank, around  £5,000- £10,000 for 6 months Money required to expand business Borrow money from friends family with lower interest rate in long-term Cash flow interrupted Raising money from banks or friends in a short term The limitation in this business plan is no registered for VAT in software SimVenture. Company decides do bookkeeping in-house to saves cost. Company credit control is under 30 days for deports. If customers credits are overdue one or two months, managers will make a phone call first; if it is overdue three months, company will utilize legal threat in the county court. REFLECTIONS ON THE BUSINESS PLAN Through creating the business plan, I learnt and understood three core points during the process of project. It has been explained in this business plan reflection. Initially, I understand that everything has been relative through the approach of functional strategy (Kazmi, 2008:435) in the business plan. Any analysis could not be totally separated into specific objective during considering of all information. For me, the first significant experience was to learn the way of consideration from the overall point of view. Previously, I thought the plan could be distinct from marketing, operation, HRM and finance aspects, same as their subordinate factors. Later, I noticed that, in operation section, I had to ensure the product design and how many components required. Then I can select SCM strategy, improving productivity, and setting price based on gross margin according to the fixed cost. I realised all elements were strongly associated by business activities. Having reading extra references (Quran: Surah Al Hajj, verse 22:76), now I believe that everything happens for a strong connection and for a reason. Additionally, I have learnt that nothing can be absolutely identified as good or bad. Daft (2012:191) recommends the advantage of start-up business is that entrepreneurs are able to create and grow the business in their own way. It is similar as traditional Chinese medicine. One herb can save peoples life, also can be poison capturing their life, which depends on the approach of using it. This understanding is essential to me as a practitioner. As a next step, I need to arrange my planning structure more strategically and comprehensively. Secondly, several of information could not be predictive in the business plan due to the special marketing period and the PLC and timelessness. As stated by Romano and Ratnatunga (1995), small and emerging firms face marketing critical issues for their survival and growth. For me, the meaningful aspect is the awareness to economic issues in the world and the PLC period in the market. In the Simventure, it does not need to consider the real global environment, at the same time, managers not need to care about more than one competitor. Although, the simulate system makes the process of business plan become easier, in the real world, those aspects can become core issues and hard, especially when the company attempts to expand global markets. However, I am conscious of the marketing information and products analysis has timeliness, which has been strongly related with global economic events. Nowak and Washburn (1988) also prove that timeliness is one of core elements focused by marketing research clients. Therefore, in the next time, I need to do more researches and consider about business environments by timeliness in the real business world. For me, the most meaningful experiences are the self-education that, as a top manager, to put all the information together, and maintain a big picture view while concurrently keeping an eye on the details (Ferrell Hartline, 2011:256). Previously, I read various theories and references based on trail Simventure. It covers the knowledge about marketing management, human resource management, financial management, international management, and management process, etc. Subsequently, the crucial information are selected and organized. Therefore, I understand, the process of doing assignment has specific procedures. Firstly, it needed to unearth modules of process, and determine the objective, mission and goals. Next, I identified core issues by functional objectives. To the question, I went to research required references and resources, providing constructive business plan. Finally, I will evaluate and prove decision-making strategies in management reports. Equally, this could be related to all project processes, not only in the business area. For example, when I map out any new field, I can go through the process with to identify goals, outline the targeting vital objectives in new field, and then to evaluation outcomes. Having this project, I found the key to acquiring proficiency in any task is the way how to study, in which, more is self-educated with unforgettable experiences. In the next project, I need to balance the macroscopic and microcosmic views with a lucid outline. Overall, I reflected three central points of building business plan. Firstly, all objectives have a relative connection by reasons, which could not be clearly separated. Secondly, business strategies and implementation have timeliness in the entire process. Finally, self-education is the key to open every field, which also require maintaining a big view with an eye in detail. THE MANAGEMENT ANNUAL REPORT OF 643744 COMPANY Executive summary 643744 Company is committed to operating computer business in the world widely, focusing on products design and production in professional services marketing. It has been established since January 2013 for three years, with capital  £10,000. Companys vision is Walking in the forefront of the times. 643744 Company aims to be a preeminent building block supplier of fashionable computer in the global market. Managers set up their mission and long-term goals for three years running business, aiming to improve innovative products and services, stepwise achieve profitable growth (Reading, 2002:360). Generally, the main achievement of 643744 Company is by product design, marketing promotion and business forecast, which follows JIT management process theory. In finance, at the end of 2015, the company has value  £1,469,003.6177, net profit  £21,925.8363, current liability  £112,245, and no long-term liability. Moreover, 643744 Company was invested by grants for  £1000 on employees training and research. By financial analysis of 643744 Company, in 2015, its ratio of ROCE (Return on Capital Employed) has achieved 59%, therein  £167,671 current assets, and  £282,562 current liabilities. Its current ratio, for measuring liquidity of company, is 3.43. The asset turnover, for measuring utilization of companys assets, is 3.51. In sales and marketing, 643744 Company applied marketing strategies (Ferrell Hartline, 2011) and international management theory (Deresky, 2010), with various marketing promotion and sales channels, resulted in brilliant corporate reputation in the international marketing. Over three years, the total investment of marketing advertising is  £47,175, including newspaper, TV or radio, direct marketing, the public relations building, business club, premium websites, exhibition, and relative premises. It goes via different sales channels, containing traditional sales, personal selling, and distributor sales. However, the company won a wonderful reputation by prestigious advertising. Through three years, 643744 Company sold approximately 3400 items, which has exceeded 1000 units than business pervious plan. In operation, in 2013, 643744 Company applied operational management strategy (Brown, et al., 2005:67), involving in SCM, JIT, PLC theories. According monthly customer research, this suitable design keeps meeting customers needs, and no need to reset. The fantastic products of professional serves completes with 79% quality, 47% performance, 79% features, and 97% style. It totally requires with 65 components. Additionally, the company offered excellent reasonable price with 30 day credits, and discount pricing promotion, which is based on Jobber Fahy (2009:204) pricing strategies. It is satisfied by customers as well. In organization, during those three years, the organization of 643744 Company expanded maturely, which followed organizational management strategies (Daft, 2012). At the end of 2015, the company had three workers, except business owner, Ian and Mark. Business owner went to full-time job in 2013. Additionally, the companys premise started at a small business centre, then changed to retail unit and the business park. Overall, 643744 Company managers realized several issue about the company premises arrangement, financial cashflow risk management, marketing promotion selection and implementation. Firstly, it is difficult to calculate the utilizing space of premises for running long-term business, due to unexpected growth of product sales in a started-up business. Additionally, credit control is a key factor in finance for a successful small company, which related its monthly cashflow and bank balance. The shortage of cash influences on SCM, resulted in stopping components delivery. Otherwise, the company will lose all of orders and customers in next month. Especially, once the company entre into globalization, it will be more difficult to collect debts. Finally, the marketing promotion has plenty of methods. Not all of them fit professional service, which needs to be measured and researched by managers experience in marketing. Eventually, even though, in the simulated Simventure, this company not needs to be registered for VAT, in real world which has to be forecasted. It is significantly important in the international business, particularly on export products and import components. The first year management annual report Business details The 643744 Company Inc. is a small new company, providing computer design and production in the world widely. In the 2013 financial year, it achieved profits with  £2,661 by  £10,000 capital. Companys mission and goals: Mission: Companys products must be sophisticated in quality, offer good value, beautiful artistic style, and philosophically inspiring. As a vigorous part in the market, company has capability improve people lives through their use of companys products. Goals: Company devises the idea design to meet customers needs and compete with competitors, And accomplishing 50 orders and 300 products of product at end of year. Key staff members Company owner goes full time in September 2013, at the end with  £1,300 on salary. Sales and Marketing During running the business of 643744 Company in the 1st year, the core issue involved in the sales that did not catch the predicted value and delayed the business planning goals. Additionally, the scenarios of the business was making business managers concerned, which increased their stress level and declined the efficiency of organizational work. This was caused by business performance instability that presented in the lack of order and sale. This problem threatened the companys reputation that is the most valuable assets for a film (Haywood, 2002:112). However, the company decided to implement marketing strategy, as below, for enhancing the sales and orders to meet the planning profit. The certain solution of symptom has two approaches. These are basic on the marketing strategy Days theory (1984:102-103) in business plan (Figure 2). Internally, managers were constantly in a state of training, with no knowledge where it should be heading (It has been explained more in the organization section). Externally, the promotional and marketing research activities were developed, in order to achieve the sales quantity in short-term and long-term. This view was supported by Rowley (1998:383, cited in Mercer, 1996: 309): The ideal form of promotion is the conversation, which takes places between the expert sales professional and his or her customer. It is interactive and conversation is specific to the needs of both. Other forms of promotion, which deal in the average needs of groups of people can only hope to approximate to this ideal. Firstly, managers applied various marketing promotion to advert its products, and build companys reputation. It followed the previous business plan. Rowley (1988:384) suggests the tool of promotion mix can be advertising, direct marketing, sales promotion, personal selling, and sponsorship, which could be combinative or individually utilized. It had been applied in the first year and represented in the table 9. Through the implementation of marketing promotions, company realized the direct marketing was not effective as in the supported theory. Jobber Fahy (2009:255) claimed unlike many other forms of communication, direct marketing usually requires an immediate response. Unfortunately, due to the result in 2013, the 5 months promotion on direct marketing did not bring plentiful customers to the firm. It was proved by monthly customer research. The reason behind of it could be the organization of company was not mature enough. The total cost nearly caught the marketing promotion budget in table 3. Second one is the implement of pricing rise or fall strategy (Jobber Fahy, 2009:204) in pricing promotion. Since May 2013, company had set price at  £670 per unit with 3% discount for 3 month. Strategically, company still had  £162 interest per unit. Consequently, the number of order quantity (Finger 3) rose from 2 orders in April to 12 orders in May. Substantially, the sales were continuing growth up. Figure : The order quantity in the first year Operation In the first year, 643744 Company implemented the product design after 4 months researching and designing. The product had 79% quality, 47% performance, 79% features, and 97% style in 65 components, which had better design than in the business plan (Table 3), represented in figure 4. The Component was purchased from ProSupply by  £7.50 per item as planning. Figure : Product design comparing with competitor and panned design In SCM, company decided retaining the current supplier rather than changing another supplier right now with more discounts. The reason behind of it was the payment to supplier. Company could not pay off all of credits by 30 days, due to without enough liquidity. This potential financial risk leads to the increasing fixed cost of production. However, the operation management planned to improve it in the next year. Organization Since in February 2013, 643744 Company had relocated at Small Business Centre ( £15 per sq ft) with 300 sq ft for one year, and fortified office resources as planned in table 5. The only issue in organization was the requirement of developing office recourses and employment in organization. It was causes of the limitation on the working time and the shortage of production equipment and tools. Consequently, the productivity and working effectiveness had been impacted, as well as the tiredness of employee. However, the Company followed in the business plan, applying hard HRM model into organization. After managers solved out the limitation of company set-up, customer and employment contracts, business owner went full-time work in September. Bratton Gold (2007: 324) argues many firms neglect Human resource development (HRD), as a central, other than to keep production in shape. Thus, the vocational training was required instantly on marketing, production, sales, and research, which had implemented by  £500 training grant supported. It enhanced productivity and proved by working effectiveness in the Simventure. However, the Company plans to recruit 2 more employees in the following financial years. Finance 643744 Company had been facing a potential liquidity issue on finance in the first year. According to the first year annual report (Table 20), too lower sales and massive debtors restrained the companys cashflow, as described by Fiedler, Brown Moloney (2002:40) in the finger 5. Thus, the money of paying purchasing was limited, which impacted on operation strategy and companys profitability. Figure : The overlap of market, credit and liquidity risk (Source: Fiedler, Brown Moloney, 2002:40) For this financial crisis, the company decided to develop marketing capability and credit control, rather than increasing long-term liabilities. Due to the low profitability monthly, additional long-term liabilities only would lead company into stressed market conditions. However, the credit control has been set up, as in the business plan finance section. Consequently, the current liability was  £38,524.575 2013 with 237sales quantity, and  £2660.9687 net profit at the end of December. On the other hand, Company obtained  £500 grant on Training by this year. Managers selected to do bookkeeping in-house. The second year management annual report Business details The 643744 Company was committed to technologies and products that focused on Professional Services selling field. It united the goals of customer value with sustainable development in the world widely. In the 2014 financial year, the company had achieved  £751,628 on sales. Companys goals were to enlarge its markets, and accomplished 100 orders and 1000 products at end of year. Key staff members Company owners salary was  £1,300. In April 2014, the company employed one staff-Ian for  £14500 per year, with 40 per week working hours, and 4 weeks holiday per year. Sales and Marketing In 2014, 643744 Company aimed to expand its markets and increase their regular sales to 1000, as described in the business plan to prepare into international market. According to the last year reserved profits, it was necessary to attract more loyal customers and enlarge the profitability substantially, in order to avoid raising credit risk and decreasing portfolio values (Fiedler, Brown Moloney, 2002:38). In this case, the price discount and money off could be applied into Simventure. The company decided to develop sales strategy promotion and advertising strategy to boost sales over the short term (Gabriel, Kottasz Bennett, 2006:506). The first solution is managing price by using promotional discounting tactics. Since January until March 2014, the company had set price promotion with 3% discount on  £670 per unit for three months. According to the break-even analysis, the company could obtain net interests  £162 per unit. As a result, during those three months, the number of sales had notably boosted to above 90 orders. Moreover, the planned price penetration strategy was unsuitable at this year growth stage in PLC. Strategically, managers applied competitor-orientated pricing strategy (Jobber Fahy, 2009:196) to change base price to  £680 and provided 2% discount, which retrained for 4 months from April. Particularly, after the company changed to SourceLine supplier, the interest can reach to  £240 per unit. Consequently, the sales order reserved to 105 units in July 2014. The order quantity (Figure 6) proved that the price strategy was effective. Figure : The order quantity in the second year Secondly, the company decided to improve marketing strategy. The Company desired to enhance the visibility and good corporate reputation in a new era of computer marketing. According to the sample survey of customer research, managers analysed the efficiency of all marketing promotion methods, resulted as Figure 7. Thus, the Company directly focused on adverts promotion and premium website rather than direct marketingthat parts budget moved to advertising as well. Figure : The analysis of marketing promotion in the first year and second year Obviously, advertising promotion had more efficiency in figure 7. Hackley (2005:84) recommend advertising can swiftly build the awareness of audience. As stated by Levinson (2007:175), most small business owner never ever consider advertising on magazines, due to the lack knowledge of regional editions, remnant space, in-house agency discount etc. By provided information of advertising in the Figure 8, trade magazine had high effectiveness. Thus, the Company decided to spend more on trade magazine with  £8,400. Figure : The advertising efficiency of Professional services In this year, the company had invested  £14,110 on marketing promotion overall, which was in excess of the promotion budget around  £2,000, listed in table 3: However, those sales and marketing promotion leaded the company increase the sales. At the end of this year, the company has sold out 1127 items totally, and achieved more than 130 orders monthly. Operation 643744 Company aimed to develop SCM with JIT management process for controlling liquidity risks. In the Ap

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

bias or not? :: essays research papers fc

Bias or Not? The most political news story that is being reported on these past few weeks has been the nomination and debate over President Bush’s nomination of John R. Bolton to be ambassador to the United Nations. One of the most watched and influential news channels, CNN has been following this routine story closely. The President nominated Bolton in March of 2005 but the Senate has been delaying the vote of the nominee for several weeks. The story on CNN was headlined Vote to end debate on Bolton fails. Though the story did discuss that it was mostly the Democrats that were causing the delay in the vote the channel also stated that their were some republicans that did not agree with the nomination. The channel tried to stand by the facts but the story did seem to state that it was the Democrats that voted against closing the debate. The story did seem slanted when discussing the issue over the Democrats wanting to obtain access to information about Bolton and Syria. The channel st ated that the was good and bipartisanship until the Democrats resorted back to a partisan approach. This seemed slante because throughout the debate both the Republicans and Democrats have been resorting to a partisan approach. Despite the slight slant ness in some of the story the primary contents of the story did stick to the facts and remained bias. Democrats Mull Politicizing Iraq War Intelligence this article found on the Fox News Channel in November of 2003 is an insider story. The following story discusses a informal memo was found concerning the planning and timing of a possible investigation into pre- war Iraq Intelligence. The memo suggested that Democrats "prepare to launch an investigation when it becomes clear we have exhausted the opportunity to usefully collaborate with the [Senate] majority.† Then the story goes on further to discuss how upset the Senates Republicans were by the leak but especially by the memo itself. A republican Senator went as far as saying that the â€Å"Democrats intended to politicize what should have been bipartisan†. Such comments seemed to elude that the Democrats were at fault for thinking of conducting investigation but that was not the matter at hand. The matter was the leak of a memo was a draft that had not even been approved. Lawmaker Hopes to Open Churches to P olitical Speech this headline on Fox News refers to a bill that Representative Walter Jones of North Carolina has introduced the Houses of Worship Free Speech Restoration Act of 2005

Earworm :: Biology Essays Research Papers

Earworm: The Song That Won't Leave Your Head I woke up and I was mortified. It was the first thing in my mind when I opened my eyes and I just could not believe this silly little thing had become as involuntary as breathing. I tried another song, but it would come back without me realizing it. I walked to work and it came with me, I sat in class and it spoke louder that my professor's voice, I even took a nap and it kept me awake. I had a stupid song stuck in my head and it wouldn't go away. What is it that happens in the brain that causes this annoyance to go on for days? And why does it remain in the head even when it's driving us so crazy that we want to scream in pain? According to research done by Professor James Kellaris at the University of Cincinnati, (1) getting songs stuck in our heads happens to most if not all of us. His theory shows that certain songs create a sort of "cognitive itch" - the mental equivalent of an itchy back. So, the only way to 'scratch' a cognitive itch is to rehearse the responsible tune mentally. The process may start involuntarily, as the brain detects an incongruity or something "exceptional" in the musical stimulus. The ensuing mental repetition may exacerbate the "itch," such that the mental rehearsal becomes largely involuntary, and the individual feels trapped in a cycle from which they seem unable to escape. But why does this happen? Apparently, repetition, musical simplicity and incongruity are partly responsible for the annoyance. (2) A repeated phrase, motif or sequence might be suggestive of the very act of repetition itself, such that the brain echoes the pattern automatically as the musical information is processed. Still, simpler songs appear more likely to make your brain itch, - like Barnny's "I love you, you love me" tune - but at the same time a song that does something unexpected can cause the brain to latch on because of whatever unconscious cognitive incident occurred at that very moment. These traits of simplicity, repetition and circular composition1 are potent because we don't remember songs as one complete image, like a picture, but as temporal sequences that unfold in our brains. (3) In other words, we don't "see" an entire song in our head; instead, one image (or line in a song) triggers the subsequent one.

Kubrick Lives Essays -- essays papers

Kubrick Lives The theory of authorship as applied to film directors is a subject that is argued extensively throughout the film world. The auteur theory was first introduced in the French film journal Cahiers du Cinema. Andrew Sarris who suggested that there are a group of filmmakers who fit into this category brought the theory to America. It states that in order for a director to be considered an auteur, there must be a consistency of style and theme across a number of films. Very few contemporary filmmakers fit into this category. One filmmaker, however, expanded his filmography over four and a half decades, and created a consistent theme and style. That director was Stanley Kubrick. Kubrick was known as a very stylistic filmmaker, so a lot can be said about his film style. His use of music, however, remains the most prominent aspect of Kubrick’s film style, especially as his career progressed. He was a master at using music to evoke feelings and create tension and confusion. The two most prominent examples of the power of music occur in A Clockwork Orange, and 2001: A Space Odyssey. The first of these two films, 2001, was created like a symphony. It had an overture at the beginning, a musical intermission, and an epilogue at the end. The classical work of Richard Strauss, â€Å"Also Spach Zarathustra†, supplies the most recognizable and moving main title theme of the film. The use of this music as well as other classical works including the frolicky â€Å"Blue Danube† by Johann Strauss gives the film a flowing quality that it wouldn’t normally have. Most of the music is light in nature, which contradicts the mystery that is unfolding in space. The beautiful imagery is matched well with the images and the editing to provide an incredible viewing experience. In A Clockwork Orange, Kubrick does virtually the same thing with music, only in a darker way. In the film, Alex is given a treatment that will make him ill when confronted with violence or sex. Unfortunately for him, the films he is forced to watch are scored with Beethoven’s â€Å"Ninth Symphony†, which is Alex’s favorite music. A sense of irony and empathy is created in that by Alex trying to take the easy way out, he is forced to give up the three things he loves most: sex, violence, and Beethoven. His love of music backfires on him once again with his crooning of the song â€Å"Singin’ in the Rain.† In one of... ...d. This solidifies the theme of duality by blatantly showing us the symbols. Style and theme in filmmaking go hand in hand. Stanley Kubrick was a master of using his own personal style to create a series of works, all with very similar themes. He believed strongly in the way music can affect the development of conflict. Every film that he has directed (excluding Spartacus) has a strong music and sound design that makes it undeniably Kubrick. (He didn’t have his usual control over Spartacus). From the lush soundtrack of 2001 to the minimal soundtrack of Dr. Strangelove, we can see that every piece of music has its place and its reason for being there. Even in the trailer for the upcoming Eyes Wide Shut, you can see that music will be an important part of it. Also, because Kubrick deals with the same underlying themes from film to film to film, we can come to understand his thoughts and emotions. The troubled protagonist and the duality of self remained consistent fixtures throughout his career. By seeing these styles and themes surface in Stanley Kubr ick’s work, we can justify designating him a film auteur. It is a shame that we will not be able to watch him grow any further.

Attention Deficit Hyperactivity Disorder

A classroom that includes students diagnosed with ADHD needs to include effective intervention strategies. According to the American Psychiatric Association, Attention Deficit Hyperactivity Disorder is characterized by pervasive and developmentally inappropriate difficulties with attention, impulsivity, and hyper activity. These students frequently have difficulties following classroom rules and can show aggression, they are also more likely to be rejected socially and have more difficulty getting along with other students. School-based services are usually needed as well as placement in a special education class. The implementation of behavioral strategies combined with the use of psychotropic medications can help achieve a favorable outcome and a greater chance of success. Children with ADHD have different behaviors that can prevent them from learning and those behaviors can range from being mildly disruptive to almost making it impossible for academic and social success. A variety of behavioral strategies can be implemented to accommodate each child's needs. These strategies should include some clear, simple expectations or rules that outline expected behaviors for all students. These rules should be posted and reviewed frequently as well as recognized and praised when they are being followed. When all students understand the expectations, they have a goal they can work towards. For some students having additional prompts can help them to maintain self control and achieve these goals. Academic tasks can seem overwhelming for some students and there are some ways to help these students complete their school work. One effective strategy is modifying the size of a task, breaking it into smaller assignments can help them achieve success. Gradually increasing the level of difficulty, and the time spent working on an academic activity might help some students learn self regulation. When they can complete a task they should be praised or rewarded. I currently work in a classroom where there are modifications made for several students depending on the subject being taught. One student becomes overwhelmed by math lessons and feels defeated before he even begins. His behavior reflects this when he becomes disruptive and refuses to even try. Several strategies have been implemented to help him and are currently being used with success. Presenting the work in smaller assignments has helped him feel less overwhelmed. First I make sure he understands the task and what the total assignment is, then it is broken into sections for him to choose from. By choosing the section he feels most comfortable to begin with helps promote a positive attitude toward the overall task. Combining this with positive reinforcement and praise for staying on task and not disrupting others has been successful and this student's average grade has increased by 10% since the beginning of the school year. Reinforcement based strategies like a reward system can be a very helpful tool as a whole class strategy. The desired behavior and expectations need to be outlined as well as what the reward will be. These expectations need to be individual and realistic so that the student can see progress toward the goal. This strategy can be successful when used regularly and followed through. This is a strategy that I currently work with and it brings positive results. I work in a seventh grade ASD classroom where there are clear classroom goals as well as individual goals. The students earn â€Å"checks† for short term rewards like 15 minutes of IPad use or art time as well as a larger goal of â€Å"Friday Fun† which is a free period of social interaction with classmates where they can choose a game or activity to play for an entire period. The goals are individualized and can be increased as the student's ability grows. Having a long term goal also gives a student room to adjust their behavior and make up for times when behavior needed to be corrected. Not every student earns this reward all the time, but the teacher helps the students understand where they need improvement and how together they can work to achieve their goal for the next week. Both articles discuss the importance of the partnership between parents, teachers, and counselors. All parties should have a good understanding of the strategies used at home and in school. These strategies are the most successful when they are followed through by all. Open communication is very important and allows for an easier transition from school to home and home to school. For two students in my classroom we send home a â€Å"Have a Good Day Chart† which gives parents a look at their child's overall behaviors as well as their progress throughout the day. This helps us to see what time of day may be more challenging or if a particular subject causes unwanted behaviors. If the student is on medication it can track the time of day behaviors are occurring and can help when parents speak with their child's doctor. Intervention services are essential to helping a student succeed. These providers evaluate from a different perspective and can offer additional strategies for parents and teachers The way to achieve the best outcome is for all caregivers to develop a plan that includes strategies that are both proactive and reactive. Encouraging the desirable behaviors and addressing undesirable actions help to teach self regulation. Tracking and recording data show growth and improvements. This helps everyone identify the strategies that work and where changes are needed as the child grows. These strategies should be reviewed and discussed by everyone implementing them. This is especially important for children in middle school or high school where there are multiple teachers working with the students each day. Consistent and clear guidelines can help students with ADHD be successful and reach their full potential. Attention Deficit Hyperactivity Disorder Wolraich and Baumgaertel in â€Å"The Prevalence of Attention Deficit Hyperactivity Disorder Based on the New DSM-IV Criteria† (1996) observe that ADHD is â€Å"the most common mental disorder in children† (p. 168). From a parent’s perspective, ADHD is a troubling disorder feared due to its adverse effects on the behavior of children. Wolraich and Baumgaertel (1996) further notes that the symptoms for the disorder â€Å"are usually well developed by school age† children and are also â€Å"most clearly and consistently observed in the school and classroom environment† (p.  170).For the parent, ADHD is a disorder that should be given strict consideration as it may greatly affect their child’s school performance and overall mental development. The article gives a detailed account of the history of ADHD, consistently citing earlier researches conducted in examining the disorder. The article also emphasizes the previous criteria in determining t he disorder. In essence, Wolraich and Baumgaertel (1996) argue that DSM-IV as a new criteria for the prevalence of ADHD.In their research, the authors obtained teachers who will fill-up a behavior rating scale for their students. Children from kindergarten to fifth grade in a middle Tennessee county participated in a second study under the same research. Moreover, the questionnaires used in the research consisted of a modified teacher rating scale in order to fit the DSM-IV. In the studies, a principal component factor analysis with varimax rotation was used. Wolraich and Baumgaertel (1996) found similarities among the studies and that ADHD is indeed a prevalent disorder among school children.With DSM-IV as the new criteria, the researchers confirm that the scope of ADHD is more prevalent than we may already know. From the perspective of the parent, it can hardly be denied that the behaviors of their children may reflect symptoms of ADHD in one way or another. By noting the research findings of Wolraich and Baumgaertel (1996), parents may soon realize that their children may have ADHD or show symptoms of the disorder. It may cause a reason for alarm which can ultimately result to an increase in the efforts of parents to constantly monitor their children’s behavior both at home and in school.I have learned that ADHD is indeed a serious disorder because it is prevalent among children who, in their young age, are still on the verge of learning many things which are basic to one’s life. If that is the case, ADHD poses a threat on the younger generation of learners which should be a sufficient reason for the learning institutions and key officers to assess the school services being provided and to create measures which will address the symptoms of the disorder manifested by young learners with ADHD. Without doubt, my perspective towards ADHD and those who have it has changed.The prevalence of ADHD among children taught me that the disorder is not a si mple problem which has a ready-made solution. Because the disorder is primarily rooted in the human brain, one’s attitude towards children with ADHD should not be in a harsh way. Learning to treat children with the disorder with respect and understanding is a must. If the facts presented by Wolraich and Baumgaertel (1996) are not compelling enough, I cannot think of any other compelling force to make people rethink their attitudes towards people with ADHD.In case one may wonder, the research done by the authors of the article is a credible research primarily because the authors have accurately and objectively presented their findings with no regard to personal bias. More importantly, the authors have the authority and scholastic records to back-up their efforts in the research. For one, Mark Wolraich is the Child Development Center’s director as well as a Professor of Pediatrics in Vanderbilt University Medical Center.On the other hand, Anna Baumgaertel is an Assistant Professor of Pediatrics in the same university. Apart from the thing that the authors have presented, there are still other things I would like to know. For instance, can ADHD last a lifetime? Inasmuch as ADHD can be acquired by children at a young age, does it also indicate that the disorder can be carried over to one’s adulthood or later years on life? Hyperactivity is one of the symptoms in determining if a person has the disorder.But it is certainly not the sole defining symptom in identifying traces of ADHD in a person’s behavior. If that is the case, then it is more than likely that grown-ups and the elderly may also manifest ADHD. Or is it? Another thing that I would like to know is whether ADHD can be directly cured by medicines or medical and surgical operations. Since the disorder has strong relation to a person’s brain, can doctors make use of brain surgery in order to ‘correct’ what seems to be a flaw in one’s brain and eventuall y relieve the person suffering from ADHD?Or can it be the case that continuous medication through pills or perhaps through injection of medicine in the form of fluids can result to an immediate and long-lasting cure for the disorder? Lastly, if ADHD is best manifested through the behavior of children, what can parents do in order to counter or at least lessen the possibility for children with the disorder to suddenly turn towards a hyperactive state?It seems much is yet to be explored about the nature of ADHD and those who suffer from it. For the moment, it is sufficient to know that the disorder is a serious problem and necessitates serious solutions for both parents, teachers and doctors. Reference Wolraich, M. & Baumgaertel, A. (1996). The Prevalence of Attention Deficit Hyperactivity Disorder Based on the New DSM-IV Criteria. Peabody Journal of Education, 71 (4), 168-186. Attention Deficit Hyperactivity Disorder Attention Deficit Hyperactivity Disorder (abbreviated as ADHD) is a term that used to describe a range of behavior related problems leading to lower concentration and poor attention.   It is a problem which affects many people across all ages in the world.ADHD leads to development of many learning difficulties in children since they cannot pay full attention in class.  Ã‚   While some individuals may exhibit one condition alone, other may exhibit a wide range of conditions including impulsiveness, restlessness, hyperactivity, and many others.The severity of the condition depends on the range of symptoms that are exhibited by the child.   Apart from academic problems, children with ADHD also suffer from social problems and most of them have shown inability to socialize with their peers at home or at school. The condition is also known by other words like hyperkinetic due to the common factor of restlessness which is exhibited by children suffering from the condition.The conditio n can be categorized together with other neurobehavioral defects which affects a wide range of population.  Ã‚   However, ADHD is more prevalent in children although statistics shows that the condition affects close to 5% of the people in the world regardless of their age. The symptoms the conditions are more evident in children than in adults and are mainly noted in a close supervision set up like in schools.There is also gender variation in the prevalence of the condition and it has been diagnosed twice in males and females.   In some case, the disease has been shown to be chronic where more than 45% of individuals who are diagnosed with the condition in their early age progress with the symptoms late to their adulthood. As individuals grow, they develop coping mechanisms which decrease the prevalence of the symptoms of the condition in later adulthood.ADHD presents one of the greatest academic challenges that teachers have to deal with.   Unlike normal children who are atte ntive in class, ADHD children exhibit a wide range of attention difficult which affects their academic performance.Therefore it is the duty of the teacher to come up with working strategies to help ADHD children to perform like the rest in class. There are various strategies that have already been proved quite effective in assisting ADHD children to improve their academic performance.  This paper will discuss ADHD in school going children. The paper will first have an overview of ADHD including its definition, pathopysiology, symptoms, and treatment. Then it will review the   social and academic difficulties that ADHD children goes through and finally it will review strategies that can be used by teachers to help students with ADHD to perform well in class.ADHDADHD was first recorded by Scottish physician Alexander Crinchton in 1798. Alexander described a condition resulting to mental restlessness which is very similar to the modern  Ã‚   diagnoses of ADHD.   However, ADHD w as recognized in 1902 by a British pediatrics George Fredrick.   Fredrick also defined a condition which was very similar to ADHD but he recognized it as an abnormal physical condition affecting children which as more evident in their studies (Cleave and Leslie, 2008).Extensive research on ADHD has shown that it can be caused by biological or environment factors and the combination of the two aggravates the symptoms of the condition.  Ã‚   According to Cleave and Leslie (2008), there are three biological factors which can predispose individual to develop the symptoms of the condition including;Child temperament   which   lead to   varied change in attitude and   personality Genetic factors   including inheritance of the condition   from parents   or close relatives Accidents leading to damage of the brain tissues which can be due to pre-birth problems or trauma which leads to hypotoxic damage of the brain Environmental causes have also been linked to development of the condition.   Environmental causes include a wide range of factors that compose the growth environment (Stern and Stern, 2002).There are wide ranges of environment factors which can predispose individuals to the development of the condition including the family stress, the style of parenting earlier difficulties faced in education, growth in  Ã‚   areas prone to negative behaviors like violence, peer influence, and many others. For example, research has shown that children who grow up with violent parents or alcoholic parents are prone to development of symptoms of the condition than others.In the United States ADHD has remained on of the most controversial social and medical issue.   Different stakeholders like medics, social workers, media, and others have all questioned the diagnoses criteria in ADHD with some arguing that the condition is over diagnosed by the present DSM-IV criteria.However this controversy in the diagnoses criteria stems from the systematic developme nt of the diagnoses tool keeping into consideration that ADHD cannot be diagnosed using clinical signs.

Ferdinand de Saussure Essay

†¢The origin of Language †¢Features of Language †¢Knowledge of Language †¢Linguistics †¢Branches of Linguistics †¢Approaches to Linguistic Studies †¢Schools of Linguistics THE ORIGIN OF LANGUAGE †¢The divine source †¢The natural sound source o Primitive words o Cuckoo, splash, rattle, hiss, buzz oâ€Å"Bow-wow theory of language origin.† †¢The social interaction source †¢The physical adaptation source †¢The tool-making source †¢The genetic source oChild development à ¨ Automatic oDeaf children à ¨ Fluent users of sign language à ¨ innateness hypothesis oWe are born with a capacity for speaking/using language †¢(See Yule, Ch.1; Fromkin et al., Ch.6) MAJOR FEATURES OF HUMAN LANGUAGE 1-Communicative : function of language is to communicate 2-Vocal : human vocal organs are used 3-Human : sophisticated use of language is unique to human species 4-Social : it Ä ±s used by the members of a community 5-Natural : Ä ±t is a natural phenomena 6-Changeable : it can and will change trough time 7-Rule-governed : there are systematic rules in it. 8-Product of mind : it is the product of mental operation 9-Arbitrary : no systematic relationship between form and meaning 10-Creative : language is used creatively MINOR FEATURES OF LANGUAGE 1-Language , unless recorded, flies away the movement we speak it. 2-Self-talk is a form of language 3-We talk in our minds : inner speech 4-We can not help but to process and understand what we hear. 5-Language can be used by some to xert their power over others. 6-Language is used for cultural preservation and transmission . 7-Language is varied : not monolithic. 8-The direction of changes in language is not predictable. 9-Language is personal as well as social. 10-Language is a means for mental and social development. 11-Language is a predictor of social identity. 12-Language reveals patterns of how mind works. 13-All languages have the same potential for development KNOWLEDGE OF LANGUAGE †¢Ã¢â‚¬Å"I know English.† †¢Produce individual sounds and combining them (Phonetics & Phonology); †¢Form and dividing words (Morphology); †¢Combine these words into phrases and sentences (Syntax); †¢Understand the meaning of words, phrases and utterances (Semantics); †¢Use the language appropriately in different contexts (Pragmatics). Language is an arbitrary and changeable set of vocal symbols which are cognitively formulated, and socially used for human natural communication. Linguistic knowledge is knowledge about language which is acquired unconsciously. LINGUISTICS †¢ Linguistics is the scientific or logical examination of any aspect and property of language including the influences of language on any sphere of life. BRANCHES OF LINGUISTICS †¢MICRO-LINGUISTICS ïÆ'ËœPhonetics (Fonetik) ïÆ'ËœPhonology (Ses Bilimi) ïÆ'ËœMorphology (Dil Bilimi) ïÆ'ËœSyntax (Sà ¶z dizimi) ïÆ'ËœSemantics (Anlam Bilim) †¢MACRO-LINGUISTICS ïÆ'ËœPragmatics ïÆ'ËœSociolinguistics ïÆ'ËœHistorical-linguistics ïÆ'ËœPsycholinguistics ïÆ'ËœNeurolinguistics ïÆ'ËœAnthropological Linguistics ïÆ'ËœApplied Linguistics APPROACHES TO LINGUISTIC STUDIES †¢Descriptive-Prescriptive †¢Diachronic-Synchronic †¢Reference Grammar †¢Teaching Grammar †¢Theoretical Grammar SCHOOLS OF LINGUISTICS Traditional Linguistics †¢Based on study of Latin †¢Applied to other languages, e.g. English, French †¢19th C. Jacob Grimm, Max Mà ¼ller, Henry Sweet Structural Linguistics †¢Ferdinand De Saussure †¢Linguistics features as abstract system governing speech †¢Langue and parole †¢While language is regarded as language system stored in the mind of its speakers, parole is conceived as the actual language behavior Transformational -Generative Grammar †¢Chomsky †¢Every language has a finite set of rules to produce infinite number of sentences; †¢ Phrase structure rules and generative rules; †¢Deep structure and surface structure; †¢Structure over meaning; †¢Competence and performance.