Chocolate Research Paper Essay

Chocolate is a key ingredient in many foods such as milk shakes, candy bars, cookies and cereals. It is ranked as one of the most favourite flavours in North America and Europe. Despite its popularity most people do not know the unique origin of this popular treat. Chocolate is a product that requires complex procedures to produce. The process involves harvesting cocoa, refining coca to cocoa beans, and shipping the cocoa beans to the manufacturing factory for cleaning, coaching and grinding. These cocoa beans will then be imported or exported to other countries and be transformed into different types of chocolate products. Cocoa beans grow in countries like Cote d’Ivoire, Ghana, Indonesia, Brazil, Nigeria, Cameroon, and Malaysia, but the highest cocoa producing country is Cote d’Ivoire. The production process of cocoa beans include: first, cocoa is harvested manually. The seed pods of cocoa are collected and the beans are selected and placed in piles. These cocoa beans are then ready to be shipped to the manufacturer for production. Cocoa grows in pods that sprout off the trunks and branches of cocoa trees. The pods have the shape and almost the size of football. The pods start out green and turn orange when they are ripe. When the pods are ripe they are harvested gently with machetes. Machines can damage the trees or the clusters of flowers and pods that grow on the trunk, so workers have to harvest the pods by hand, using short, hooked blades mounted on long poles to reach the highest fruit. The cocoa seeds then undergo a process of fermentation by placing them in large, shallow, heated trays or by covering them with large banana leaves. If the climate is right, they may be simply heated by the sun. Workers mostly women come along periodically and stir them up so that all of the beans come out equally fermented. This process may take up to five or eight days. After fermentation, the cocoa seeds are dried before they can be scooped into sacks and shipped to chocolate manufacturers. Farmers simply spread the fermented seeds on trays and leave them in the sun to dry. The drying process usually takes about a week and results in seeds becoming reduced to about half of their original weight. During the production process, labor is not equally divided between men and women who work in the planation; this brings us to the issue of gender division. The gender divide that exists on the cocoa plantation is that most farm work is conducted by men, although most certainly there are tasks where women are very active, such as scooping the beans from the already opened husks, turning the beans during the fermentation and drying process, and sewing the jute sacs needed for the packaging of the dried beans. Women in the farms normally tend to the needs of the family. When female labor is hired during the harvesting time the wages given to them are not the same as those for men. Perhaps another reason why men are preferred is because of their assumed higher productivity rate compared to that of women. Due to different practices followed in individual regions, even within countries, the participation of women and their assigned tasks vary enormously. For instance, because of the popular method of sun drying cocoa beans in Ecuador, it is necessary to â€Å"clean† the beans. This job is mostly undertaken by women. This is not the case in Ghana or Brazil where sun drying is accomplished while protecting the beans from foreign matters and waste. It is interesting to note however that there is no specific pattern for the assignment of tasks to women, except during the harvest when the scooping of the beans from the opened pods is primarily performed by women in most cocoa producing countries. Given the great differences in the systems of production in producing countries it is difficult to find a common percentage that reflects the average participation of the female work force. It is worth mentioning that unlike the coffee agricultural sector, there are no associations or specific groups that house ‘women only’ involved in the cocoa sector at any level, although all associations and cooperatives are open to all who qualify. Due to fluctuation of cocoa prices in the world market, farmers have no long-term security, and in some situations, they do not have enough funds to support their farming business. Cocoa farmers are always faced with financial hardship; they are not able to provide for their families as they would want nor have enough funds to start up their own farming business because they only receive a fraction of the proceeds from the selling of the beans on the world market and there are many people in the trading chain. Cocoa farmers around the world face many challenges. It is estimated that about one-third of global cocoa crops are destroyed by pests and diseases every year. Many cocoa farmers have limited access to the latest agricultural technologies or methods of cultivation and few of them have business backgrounds to help them effectively market their products and manage their operations. Many of the farming communities live in poverty and are infected with diseases. Industry groups, governments and consumers worldwide have raised concerns about the use of pesticides and child labor on West African cocoa farms. An interview conducted by Christophe Koffi showed that one major problem that women in cocoa production encounter is the lack the of financial capability or backing due to the fact that most of these women find themselves in a male dominated occupation. It is very difficult for them to secure financial aid or loans to manage their farms. For instance, â€Å"Women cannot inherit or even create a cocoa plantation under our patriarch-dominated tradition,† said Vanie, criticising what she called a â€Å"backwards and misogynist† practice† (Koffi, 2008) because we still live in a patriarchal dominated society where women do not have the right to own lands and properties. This paper further talks about the sustanability in the production process of cocoa. We will be looking at Lindt& Sprungli’s which is a family company and a major producer of chocolate and other cocoa products with a headquaters in Kilchberg, Switzerland. Lindt & Sprungli is one of the few chocolate makers that have complete control over every step of the production chain starting with the precise selection of the finest cocoa varieties from the best growing areas in the world right on through the careful and expert processing until ending with the elegant packaging. Lindt gets its cocoa beans mostly from Ghana and Central and South America. Lindt has been very conservative in the amount of energy its invests in the chocolate production process. Each existing and future facility and investment undergoes very detailed analysis to determine how much energy can be saved. Through better insulation and energy recovery, Lindt & Sprungli was able to cut down on energy consumption by more than 13% per ton produced between 2004 and 2010. The company intends to continue reducing the energy consumption rate per ton produced by an average over the coming years. According to the Lindt publication, The company’s efforts: since 1999, Lindt & Sprungli’s Swiss subsidiary, Chocoladefabriken Lindt & Sprungli (Schweiz) AG, has been an active member of the Lake Zurich Energy Model Group†¦The Swiss government and independent engineers have audited the progress and as a result, the Swiss subsidiary has been granted the official certificate. (Kilchberg, 2012) Since 2007, Lindt & Sprungli under water conservation has been participating in the Carbon Disclosure Project (CDP)†¦Since then, the measurements of water emissions and energy consumption have been largely based on the concept of the ‘Methodology Fossil Fuels’ set out in the CDP Protocol†¦Waste water, Lindt & Sprungli continuously monitors and analyses the use of water and the output of wastewater in the production process and intends to further reduce the use of fresh water in this process, which, in turn will impact the output of wastewater (Kilchberg, 2012). The major people that benefit from cocoa production are mostly the big corporations. The corporations exploit cocoa farmers in the sense that they do not pay them the true worth of their labour. Women who work on the farms are underpaid and marginalized. There is also the issue of child labor where children of school going age are forced to work on the farms instead of going to school thereby denying them the right to education. The big corporations are not forth coming in releasing information as to how and where they get their cocoa beans from. Most of these African countries where these big corporations get their raw materials from do not have access to good roads, health care facilities, schools, electricity, and there is poverty in most of the communities. These big corporations buy the cocoa beans at a very cheap rate and then import them to the western society and the refined product is processed into different kinds of chocolates (e. g. chocolate drink, chocolate bars of different shapes and sizes and chocolate candies etc. ) . Which are sold consumers at exorbitant prices considering the price at which the cocoa beans are bought from the cocoa farmers. This explains how capitalism and big corporation exploit cheap labour. Almost everyone enjoys a bit of chocolate every now and again. But if you take a closer look at how cocoa is produced, it may well leave a bitter taste in your mouth. The conditions under which the cocoa farmers in many producer countries live and work are worrying: Despite the fact that cocoa is usually their main source of income, the families struggle to make a living from it. Child labour is not uncommon. As consumers we can work with NGOs to find means of helping the farmers to adapt to new systems of cocoa farming that result higher yields, under socially more acceptable and environmentally friendly conditions, to meet market demands and hence ensure a stable flow income. As a major part of the global cocoa industry which has remained inactive and invisible for so long, consumers of chocolate can demonstrate that they want slavery in the cocoa sector stamped out, and your pressure can highlight their lack of commitment and make them more accountable. Finally all cocoa products, including chocolate, run the risk of being tainted by child labour and slavery. To achieve a satisfactory standard of ethical production in chocolate consumers must help to ensure that companies commit to credible and sufficient actions against such things as use of child labour and the exploitation of cocoa farmers and not make false and unsustainable promises to consumers of being â€Å"slavery- free†. The consumers can liaise with government bodies and NGOs to negotiate fair prices for the purchase of cocoa products and this will in turn help the farmers to gain access to basic social amenities of life. Reference Page Archer, D. (2012). ADM’s commitment to sustainable cocoa. Milwaukee: Copyright 2012 Archer Daniels Midland Company . Clarkson, T. (1998). Anti-slavery. Retrieved November 15, 2012, from www. antislavery. org: http://www. antislavery. org/english/privacy_policy. aspx Kilchberg. (2012, April 22). The environment in the Production Process. Retrieved November 13, 2012, from www. Lindt. com: http://www. lindt. com/swf/eng/company/social-responsibility/lindts-sustainable-cocoa-supply-chain/ Koffi, C. (2008, November 7). Ivory Coast women defy taboos. Retrieved November 11, 2012, from iol News: http://www. iol. co. za/news/africa/ivory-coast-women-defy-taboos-1. 423405.

Early Birds Catches the Worm

Each proverb has the features of time, and new ideas can be added to the old ones. The classic proverbs never worried about fading. Like the proverb â€Å"Early bird catches the worm†, people all over the world are still saying and using this phrase today. Although in different languages, we use completely different words, but still convey the same meaning. For example, In China, there is a similar saying: â€Å"The quick-footed mount first. † This proverb usually plays a traditional role in dally life, fitting well Into this competitive and materialistic society.Everybody wants to be the first, to stand on he top of the mountain. Although the university year is still in its early stages, students who are applying for graduate positions In 2009 have almost missed their chance. Companies have begun their recruitment earlier this year. These companies never worry about lacking in people to work for them, what they really need are those who are eagerly preparing his/her re sume to get into the company. Human life Is transient, so beginning a career at a young age is an â€Å"early-bird† and wise choice.American's Next Top Model has been a hot TV show for years, which I have been watching for a long time. Every time the contestants are surely to be at a young age, because if you get into the competition at 25,you are already too old and the Judges and other competitors will always use age as an issue, though unconsciously. Most people have their dreams when they are only children. And the dream remains In their minds as time goes by. Take action, be the early bird, even with mistakes along the way. Although some of them seem so far.Don't worry, there is also a saying: Clumsy birds start flying early. Upon closer analysis by changing or adding a few words in the proverb, a whole new perspective is shown: â€Å"Urban bird catches the arm. † Early birds may catch the worm but city birds prefer a little bit late. They don't rely heavily on wo rms from the trees. Instead the city birds visit bird feeders later in the day. The researchers believe the difference in feeding is due to the â€Å"urban heat island effect†, as the city becomes warmer, the birds don't need to get up early to replace energy lost overnight to keep warm.Within a word play, the anta-proverb has similar pronunciation as the original . Using the old frame is a good way to attract other people's attention, in a humorous demonstration. Let us not focus solely on the costive side, the usage of this proverb has a great limitation. Everything in this world is not absolutely right. Getting up early means less sleep. Recently research shows that â€Å"Night owls rule the roost,† scientists found out that the late risers tire less quickly than those who get up at the crack of the dawn, thus they could have more energy to do their work.To think more critically, the early birds catch the worm, and the early worms get eaten by birds. Do any of us wa nt to be the victim? It is easy to market's need and thus become the sacrifice. Coming late into the business world ND learning from the established competitors can offer an advantage. Rival birds can turn up pretty much whenever they want and still get a decent feed, as long as they hit the right spot, to say, there are plenty of worms hanging around everywhere.Here is a interesting one: â€Å"The early bird would never catch the worm if the dumb worm slept late†, the phrase also shows not every early-birds choice could be success. There are risks we need to undertake. In any cases, getting up early is often unnecessary. â€Å"Early bird† suggests that we should be more diligent in achieving our goals and desires. Taking more time to think, the proverb â€Å"think before you leap† always relates to the early bird wisdom. After comparing this proverb and illustrating so many examples, the proverb reminds me of the Darning's famous theory, Survival of The Fittest .Although the â€Å"Early bird catches the worm† need context to make it more conceivable, if all the people on the earth were willing to be the early birds, the scene would be chaos. The nature order among human competition, allows a functional world. The fact is early birds eat early worms, late birds eat late worms. Early bird catches the worm, this proverb impresses me so much is because mama always using it to wake me up, reminds me of home. That's the warm and positive side for me. There are so many things to list within one proverb.Every coin has two sides, I don't like to live in a competitive world, but welcome to the reality world, I need to face it whether I like the life style or not. Just to say I will never treat it as my lifetime guideline. To grow with a ordinary heart, to live each day with energy and happiness is the best for me. Find out what suits you best, that's the most important essence we could characterize as our own unique proverb. Citation: The Cou rier Mail. Night owls who rule the roost†. 25 April 2009. Lexis's.

Losing Weight: Lifetime and Commitment

Dieting Is a hard task to do If you do not have the right tools to accomplish It Besides there is so many diets and exercise regimes out there in today's world that it is enough to confuse a person. It Is going to be a challenge and a personal goal for some people. How about you? Do you want to make a life time change to get your body, mind and soul on track and plus keep off for life, too? It is all about making a life change in helping people get control of there body and emotions and well-being.Many overweight people have other deficiencies in there body for they are not eating he right foods and getting the right nutrients to satisfy there bodies. They are usually nervous, irritable, moody, depressed, and feel sluggish all the time of which makes them not think clearly and have little or no energy to do there dally tasks In life. When people do not have a balanced diet and exercise regime they are constantly hungry and feel sluggish. Remember you have choices In life so make the right choice.By making a life time change and commitment to change bad habits and exercise will help anyone have positive results as live longer and maintain weight. First, you need to have a goal as to how much weight you want or need to lose. So you need to set the goal but do not set a time limit and give your self adequate time to reach it. Take as many steps as possible to reach that ultimate goal. You and everyone else can do this if you Just keep your eyes focused on the goal.People should really go too quiet place and really search down deep in side your self if you have the guts to do it or not and the commitment in doing this for there body and mind and soul. So think about if you really want this and then go do it for yourself. It would not hurt togged kind of a support team to help keep you titivated and then when It gets rough sometimes and you want to quit you will have this and won't give up. You could go to friends, loved ones, churches, diet groups and maybe come up with one but you do need to choose one. Choosing one will help give support when needed and enjoyment through your program that you chose.Rather a team Is better than one to help you reach your goals In life. Secondly, you need to choose a diet and exercise program to start to reach that ultimate goal. To find the right diet take a look at your body and analyze it to see what you need to work on In order to reach your goal. Do some research on it. Like look up different diets and chose one and same goes for the exercise program It Is actually what ever fits that person but to tell you the truth I personally found a diet after doing the Atkins Diet that really works and keeps you satisfied all day.I found that the Atkins Diet was high in fats that can raise your cholesterol and lead more to heart disease. It would keep your blood pressure up and definitely is not good for your body. We all know that none of this Is good for they don't care about there persons or natural sugar that o we not harm you. These are suppose to work together for you to lose weight. Len the Atkins Diet you only lose up to approximately 10 lbs. A month compared to the Venetian Wellness Diet.The Venetian Wellness Diet where you can eat all day long and feel full and not be Irritable. It Is all with good wholesome food and filling content with it. You still eat protein, veggies, fruits, and not breads of pasta or rice not unless It Is brown. Most of the foods maybe organic It will give It a more natural 1 OFF feeling and better tort you to eat. The change is g tort the wellness tort your body. You limit your dairy and your portions are a filling size. You are choosing your foods in a differently manner.The pounds will soon start slipping off and you will start feeling different inside to and with incorporating a exercise regime into helping you lose weight you will be helping complete your goal. You might want to do some more research on exercise you are going to chose for you self to do. Maybe you can do aerobics, swim aerobics, walking or doing a little of everything put together at least three times to five times a week. So with these tools you will be able to keep the eight off and get there body in shape.Finally it is all about making a long life commitment and life change for yourself. The Venetian Wellness Diet will do Just that for you. You can eat all day and not feel hungry and with exercising you will have all the tools you need to achieve that life time goal and commitment for life. All of this will get your body in shape emotionally and psychologically balanced in your life. So lust don't think you will have these tools your whole life to use and that meaner that you are making your body balanced and ready to fight the world out there with your new life commitment and change.After using this diet and exercise you will have a new level s of well-being in your life and energy you thought you never had before. With the Venetian Wellness Diet and the exercis ing regime that you chose you will experience this only if you commit too life change . Just watch you transform and with these tools you will be able to reach your goal. You have to be religious and passionate about yourself to do this. So make that life time long commitment and change to change your body for yourself and complete that goal. Get going now and make that change and commitment in your life for a lifetime.

Julian the Apostate, Constantius II by Ammianus Marcellinus Essay

Julian the Apostate, Constantius II by Ammianus Marcellinus - Essay Example Ammianus Marcellinus was a historian with a great sense of things. For someone who has live through the times of rich and vivid events, nothing is expected except a picturesque account of events. However, in the light of what the term history meant, accounts such that of Marcellinus may be carelessly considered as merely a narrative of personal experiences and commentaries. His writings may have the patches on; there may be biases on the accounts he had written. When confronted by events and circumstances, it is inherent for every person to choose our sides, to come up with our foregone conclusions. Historians are not excluded. For a case in point, consider his writings on Julian the Apostate and Constantius II. In such situations, there is no better way to come up with a judgment than to fairly analyze the condition and consider the circumstances. Did Marcellinus have his partiality in writing the accounts of these two figures? Moreover, did he, in terms of politics and religion, fa vor the system of Julian the Apostate over Constantius II, and have it reflect in his writings?Most accounts tell that there is not much known of Ammianus Marcellinus’ life. From sources, it is said that he was born to a noble Greek family in Antioch, Syria, about 325-330. There are no accounts of his early life and educational background. Most of the biographical information available tells that he entered the Roman army at a young age, at the time when east had Constantius II as the emperor.

A specific function of an element of the U.S.Department of Homeland Research Paper

A specific function of an element of the U.S.Department of Homeland Security...I want to ICE - Research Paper Example However, it seems that many had barely looked at the surface, while credits for the many accomplishments in the preservation of national security were enjoyed by renowned groups only. There is more to ICE than meets the eye. Crime has many forms, and until one is committed, authorities may not be instigated to design the appropriate laws and the organization to enforce them. The establishment of the Immigration and Customs Enforcement (ICE) entails a long history of trial and error. From a small unit, it evolved into an all-encompassing crime-busting organization, and front-liner in the operation against terrorism. In fact, before 9/11, immigration and customs authorities were not widely recognized as an effective counterterrorism tool in the United States. ICE changed this by creating a host of new systems to better address national security threats and to detect potential terrorist activities in the United States. ICE targets the people, money, and materials that support terrorist criminal activity (Fay, 2007, p. 591) So much was the impact of the 9-11 tragedy that the Government addressed areas of â€Å"national security† before all other concerns. Intelligence planned to engage in strategies to monitor all means of communications and dissemination of information. The President made efforts to build close relationships with his counterparts abroad to extend the nation’s scope of alliance. Federal authorities created ICE to ensure that whatever the cause of the blunder was, it will not happen again. Today, although ICE is hardly ever portrayed in the big screen like the FBI and CIA, and thus is not identified by many, it is always included in the cast when national security is at risk. They perform such functions that affect millions of lives, directly and indirectly. Since then, the United States has not been subjected to anymore instance of terrorist attack, and the question lies on whether it was due to ICE’s effective performance, or

Technology impact on the organization Research Paper

Technology impact on the organization - Research Paper Example Factors of production include the inputs required in the production process in order to accomplish an organization’s objective in achieving its desired output. The desired output may be a finished product or a service depending on the organization. They include labor, capital, land and enterprise or entrepreneurship (Lee & Edgar, 2002). This makes technological progress the mover of development in the economy. Labor in an organization is termed as either skilled labor or unskilled labor. Most organizations will prefer the use of unskilled labor as much as possible since it is cheaper as compared to skilled labor. It is also in many instances readily available. On the other hand, skilled labor is needed, especially where technology brings in the aspect of automation in processing. Capital is the initial amount of money required to carry out organization affairs. Depending on the investment targeted, an organization requires variant amount of capital. An organization with an automated computer controlled process cannot have the same amount of investment with another of the same kind. Initial cost of high technology is high, but this ends up becoming economical due to efficiency and high productivity in processing. This is a basic factor that deals with the management of a company. Workers in the managerial positions and are conversant with high grade technology know how to bring out better results. This brings in the aspect of â€Å"Just in Time† delivery of goods and services. One of the roles of technology includes increased productivity of the workers. This is enhanced by the use of efficient machines in operations, thus making it possible to exploit the worker’s potential. There is optimal resources utilization; this includes space, time, process, human resource among others. When there is no waste in a process, optimal profits are obtained, and making profits is the main goal of any business organization (Montano

Charlene Barshefsky Case Analysis Assignment Example | Topics and Well Written Essays - 2000 words

Charlene Barshefsky Case Analysis - Assignment Example Using analytical framework to view the key issues and complexity of the case, the researcher will try to justify Charlene Barshefsky’s actions and suggest some alternate approaches to view the problem. This case is all about complexities regarding trade and cultural negotiation process hence the researcher will not try to give any subjective viewpoints which might spoil the purpose of the case. Recognizing multiple aspects of the case such as coalition building approach, bilateral dealings, â€Å"barriers† to agreement etc will be key features of the discussion. This piece of work will also try to understand the actions taken by respective authorities in the case in order to facilitate protection of intellectual property rights (IPR), improvement of IPR policy of People’s Republic of China and address human right issues. In the least part of this paper, the researcher will summarize the personal learning output from the case. Question 2 Research scholars such as Shapiro (2001) and O’Donoghue and Zweimuller (2004) have classified IPR as a pretty much sensitive issue when it comes to negotiating between countries. The case sheds light on International trade negotiations between China and USA in context to intellectual property rights (IPR) violation. According to the case, during mid 1990’s, USA accused China for its poor standard of IPR law which gave the opportunity to pirates to counterfeit and illegally copy entertainment and software products of USA. In 1993, USA representatives reported that Chinese pirates counterfeited USA entertainment products such as DVDs, movie CDs, software and exporting pirated CDs, DVDs to South Asian and North American countries. Such level of piracy and counterfeiting had negatively affected the trade relationship between USA and China and also decreased the market share of IPR products of USA. The case is presented from the perspective of Charlene Barshefsky who was then then-Deputy United Stat es Trade Representative (USTR) and was responsible for handling the negotiation process. In the first part, the case discusses about the challenges and barrier of negotiation faced by Charlene Barshefsky while the second part of the case study, discusses about strategic intervention used by Charlene Barshefsky and her team members to deal with the situation. However, the case is more about trade agenda regarding cross cultural negotiation process and intellectual property right issue rather than description of tariff and non-tariff obstacles related to free trade. According to the case, key assignment for Charlene Barshefsky was to offer or formulate viable strategy for establishing new intellectual property rights agreement with Chinese government. The case also underlines alternatives such as initiating Section 301 which is a unilateral action and multilateral approaches such as working as joint force with other countries to improve the IPR standard in China. Question 3 Hulse and Sebenius (2003) pointed out that working as USTR was a challenge for Charlene Barshefsky which she had taken in positive manner. She had understood the fact that she could not get the support of USA government unless make the IPR violation in China as domestic issue of

Assessment of parental satisfaction with dental treatment under Thesis - 1

Assessment of parental satisfaction with dental treatment under general anesthesia in pediatric dentistry - Thesis Example In order to provide health professionals, health plans and health care providers, purchasers, integrated delivery systems and others with an comprehensive and accessible tool that will help them obtain objectives and exhaustive information on evidence-based clinical practice guidelines, a complete database called The National Guideline Clearinghouseâ„ ¢ (NGC) was initiated by the Agency for Healthcare Research and Quality (AHRQ) of the U.S. Department of Health and Human Services which was formerly created by AHRQ in collaboration with the American Association of Health Plans (now Americas Health Insurance Plans [AHIP]) and the American Medical Association (NGC 2008). One of the guidelines presented in the NCG is the Guideline on Behaviour Guidance for the Paediatric Dental Patient published by the American Academy of Paediatric Dentistry (AAPD) in 2006. This guideline was the result of the symposium held by AAPD in 2003. Behaviour guidance is based on scientific principles, which requires communication and interpersonal skills such as empathy, coaching and listening. The guideline states the importance for dental practitioners to have acquired adequate knowledge and skills all through dental education regarding the effective treatment of childhood dental diseases. Careful and effective handling of these diseases systematically requires modification of the behaviour the child under treatment. This practice is referred to as behaviour guidance which is a continuum of interaction engaging the dentist and/or dental team, the patient and the parent/s bound on the subject of communication and education. The goal here is to advocate a deeper understand ing of the importance of good oral health and the relevant procedures undertaken for this to be accomplished and, in the process, assuage dental care apprehensions and anxieties. Another guideline presented in the NCG is the Safe Sedation of Children Undergoing

Risk Management Essay Example | Topics and Well Written Essays - 1000 words

Risk Management - Essay Example This paper will describe a risk context that may be faced by the top level executive of a bank while marketing it financial services. Risk contexts A bank executive normally faces different types of risks once the bank deals with ranges of transactions and uses a large amount of leverage every day. When a bank’s financial position becomes weak, naturally its depositors may withdraw their savings. Under such a difficult situation, the bank cannot sell debt securities in financial markets; and this condition would worsen the bank’s financial state. For instance, the major cause of 2007-2009 credit crisis can be attributed to the fear of bank failure. According to Pyle (2007), although a bank executive may share many of the same risks of other organizations, the major risks that really trouble an executive are liquidity risk, credit default risks, interest rate risks, and trading risks. Risk Identification and Analysis 1. Liquidity risk In case of a bank, the term liquidit y indicates its ability to pay bills and other payables, to repay money to a depositor, and to lend money to a borrower as part of bank’s credit policy. Hence, liquidity is the basic tool that is used to assess the financial viability of a bank. A bank executive faces great troubles while dealing with liquidity management because demands for funds are often unpredictable. Other off-balance sheet risks including loan commitments, letters of credit, and derivatives also constitute liquidity risks. A loan commitment indicates a line of credit that a bank issues on demand. Letters of credit are credit securities by which the bank guarantees that an importer will pay the exporter for imports or a commercial paper of bonds issuer will repay the principal. Finally, derivates are also an off balance sheet risk, which played a crucial role in the collapse of American International Group (AIG). 2. Credit Default Risks Credit default risk occurs when a borrower fails to repay the loan a mount. In general practice, loans are written off after a period of 90 days of nonpayment. Law demands banks to maintain a loan loss reserves account to cover the losses arising from unpaid loans. A bank executive or manager has the responsibility to ensure that the borrower has submitted collateral securities that are adequate to cover his loan amount. In addition, the bank executive has the primary responsibility to recover the loan amount from the borrower. Therefore, bank executives would be liable to answer the board of directors when a loan goes unpaid. 3. Interest Rate Risks Banks usually pay lower interests on its liabilities such as deposits and borrowings and charge higher interests on their assets such as loans and securities. Hence, it is obvious that difference in these interest rates is the main source of profit for any bank. However, a bank’s terms of liabilities are usually different from its terms of assets. In other words, interest rate paid on liabilities i s highly subjected to short term rate fluctuations while interest rate earned on assets is fixed. Sometimes, the interest rate variation may cause the bank to pay more for its liabilities and thus reducing the bank’s profit rates. Under such circumstances, a bank executive faces interest rate risk. Since the interest rate fluctuations are unpredictable, often a bank executive f

Designing VB Applications Across Multiple Platforms Essay

Designing VB Applications Across Multiple Platforms - Essay Example It enables the programmers to create a source code that is specific, easy to read, and explicit. Some of the advantages of implementing the naming convention system are: It supports the use of the tools used for searching and replacing, which has minimal possible of incurring an error. It makes it clear for one to understand the source code since ambiguity will is made minimal. It gives the source code a look that is professional-like. It also gives the code an aesthetic look. So we can conclude that the naming convention beautifies the code. It facilitates consistency, hence enabling the readers to base their focus on the content of the source code, rather that its layout. In the code, the content as opposed to the layout is the most important. It gives extra information about the source code, hence making it easy for the reader to understand the code. This additional information makes the code to be self-descriptive. The user is able to understand the function of each line of code. One is able to make amendments and maintain the source code with ease. It makes it easy for other programmers to easily understand your code. This enables other programmers to be able to upgrade your code with ease in future. ... The unit is supposed to come last, starting with an underscore and without being cased. The use of the Option Explicit. This statement entails the declaration of all variables used in the Visual Basic code. Declaring of variables helps in saving on the programming time, by minimizing the amount of bugs caused as a result of typos (Snell & Powers 2010). The following steps can be used to create a calendar scheduling system: On the Start Menu, Go to All Program, then the Windows Calendar. Click on the New Task Button, or the New Appointment’s Button. This enables one to start on a new and an empty appointment that one can add on their Windows Calendar. One is now able to add information on the new appointment. This is done on the details pane. This is where one specifies whether they want the Windows Calendar to remind them about the task. Click anywhere outside of the Appointment’s Pane to view the task that you added to the created appointment. The View Button enables o ne to make a cycle throughout the views. One can be able to select the required view from the sequence of views. A Calendarscope is a calendar developing software that enables one to plan, manage, and schedule special events, for instance meetings, appointments, among others. It enables one to view all their events on a daily, a weekly, a monthly, or a yearly basis. One can be able to get a quick glance of one day using any view of the calendar. Using this software, one can be able to create single or frequent responsibilities and events, set a reminder for events that are upcoming, put different colors for the different types of items, among other things. It supports many features, among them the drag

Marketing Strategy of a new energy drinks Essay Example for Free

Marketing Strategy of a new energy drinks Essay All around the world drinking beverage is like an entertainment for everyone. Millions of people have beverages everyday for refreshment. Our report is about the Energy Drinks of Bangladesh. In this report we have explained and analyzed how some of the five top selling energy drink companies are operating in Bangladesh. Energy drinks are becoming a major part of the beverage industry due to the rapid increase of their market competition and consumer awareness. In Bangladesh, Tiger and Speed are the most famous amongst all other energy drinks. Most of the energy drink companies act and carry out their marketing strategies in similar ways. More innovative steps are required to improve their market share. The goal of this report is to find out about the present condition of energy drink market in Bangladesh, major competitors we are likely going to face and how we are going to overcome the challenges in the market when we will launch a new energy drink named ‘Spirit’. With the use of effective marketing mix and other strategies we are going to target the young people of Bangladesh to sell our product. We have done a clear competitive analysis to take advantage of the weaknesses of our competitors and opportunities of the market. We are likely going to invest hugely in the beginning of launching our product to inform our customers about our product. As a new product company we are promising our customers to offer a unique product which will give them energy along with the excitement to enjoy their lives. Section 1: Current Market Analysis 1. 1: Top 5 brands in the market (how we find it) –We have surveyed in some of the areas of Dhaka city, like – Farmgate, Motijheel,Uttara, Gulshan; to find out the names of five most top selling energy drinks. According to the survey we have seen – Tiger, Speed, Shark, Sting and Boost are most popular in the energy drink industry of Bangladesh. We went to different departmental stores as mystery shoppers and randomly asked about the drinks each shop offers and found out the names of the energy drinks customers tend to buy more. 1. 2:Product Analysis -Product Level – Product leveling is the first and most important step taken by product planners to add more customer value to their products. It includes 3 main levels: Core Customer Value, Actual Product and Augmented Product. Core Customer Value – The first thing that comes to everyone’s mind when they talk about energy drink is ‘source of power’. In Bangladesh, all of the energy drink companies basically have the same core customer value, which is mainly about replenishing the energy of their customers through providing them with their energy drinks. However, different brands use different mottos to establish their customer value. For example: Shark’s motto says, â€Å"Real Energy, Real Power†; in other case Speed’s motto says, â€Å"Heavy Energy†. This way each products motto varies from each other and creates a different view about the product in their customers mind. Actual Product – This level includes Brand name, Packaging and Product Attributes. Under Product Attribute comes: Product Features, Design and Quality of the product. i. Brand name: A brand name is an identity of a product, without it no product will even exist. Each brand name establishes their uniqueness with their different names, signs and logos. All of the energy drink companies have a brand name of their products, which differentiates each of them from their competitors. The names ‘Royal Tiger’, ‘Boost Energy Drink’, ‘Speed’, ‘Shark’ and ‘Sting’ – each of them stands as a brand name for their product, marketed by different producers. ii. Packaging: Packaging describes the way a product is protected using a container or wrapper and at the same time provides all the required information regarding a product. The energy drinks – Sting, Speed, Shark and Tiger are all available in both cans and bottles. Only Boost is available in steel can in the market. iii. Product Features and Designs: Features explain the tools which enable a particular energy drink to gain competitive advantage over other energy drinks. The tools include – design, smell, taste and color of each energy drink. The designs of Sting, Speed, Shark, Boost and Tiger energy drinks are more or less the same. They are mostly available in two forms and in two shapes, one in a round shaped plastic bottle (250 ml) and another in cylindrical steel can (250/270 ml). They all taste sweet, expect from Tiger and Boost; which tastes sour as well. iv. Quality: Speed, Tiger, Shark, Boost and Sting have been maintaining a good standard of quality by providing the adequate amount of energy to their customers and helping them to deal with strenuous activity. However, no matter what the quality level is; over consumption of energy drinks can always prove harmful for human beings (even though they might be alcohol free). Augmented Product– There is no after-sales service, delivery service or product support for these perishable products (like – energy drinks). Labeling– Labeling is the most important part of every energy drinks. It informs about the ingredients of the product, who made the product, safety issues regarding the product and the web address of the company. Speed, Tiger, Shark, Sting and Boost all maintain an informative and trendy labeling to help their customers to make the right choice. Their ingredients contain Carbonated Water, Carbonated Sugar, Vitamins, Preservatives, Colors, Antioxidant, Sodium Benzoate, Ca-D-Pentothenate, Citric acid, Tartrazine Yellow and many other things according to the standard or permissible level. They even mention on their labeling that, it can be harmful for children and pregnant women. However, they are manufactured by different companies. ‘Speed’ is manufactured by Food And Beverage Limited, ‘Shark’ is manufactured by Global Beverage Company Limited, ‘Sting’ is manufactured by Transcom Beverage Limited, ‘Tiger’ is manufactured by AST Beverage Limited and ‘Boost’ is manufactured by Boost Drinks Limited. Brand Positioning – One of the challenging things for the energy drink companies to do is, to create a permanent place in the customers mind about a particular brand. To do this, all of these energy drink companies need to locate their brands at 3 levels – i. Product attributes ii. Benefits iii. Beliefs and Values. i. Product Attributes: Even though Tiger, Speed, Shark, Boost and Sting all have the common color of the drink yellow; their smell is very different from each other. Boost tastes sour, Tiger tastes both sour and sweet, Sting tastes like strawberry and both the Shark and Speed taste sweet. ii. Benefits: The common benefit any consumer can gain from having an energy drink is, ‘spiritual force’ which will make them more active in their daily life. Still, every energy drink has it’s own way to inform about their benefits to their loyal customers. ‘Royal Tiger’ benefits their customers by recharging their energy physically, ‘Boost’ helps their customers with being more efficient, ‘Sting’ provides their customers with vitamins and other nutrients, ‘Shark’ strengthens the stamina of their customers and ‘Speed’ increases the concentration and performance level of the individuals who are consuming it. iii. Beliefs and Values: Satisfaction comes from the core belief that consumers hold in their minds about a certain product. Customers believe – ‘Sting’ and ‘Tiger’ will give them satisfaction with better taste, ‘Boost’ will boost their energy, ‘Shark’ will give them real power and ‘Speed’ will help them gain extra speed. This way customers share their different beliefs and value for each energy drinks differently. 1. 3: Price Analysis– Size of the Bottle Product Name 250 ml 270 ml 330ml Speed 25 BDT Shark 25 BDT Sting 25 BDT Boost Tiger 25 BDT Size of the Can Product name 250 ml 270 ml 330 ml Speed 30 BDT Shark 35 BDT Sting -Boost 170 BDT Tiger 30 BDT 1. 4: Promotion Analysis Promotion : Advertising is the most crucial part of a company’s marketing mix. It is the first step to inform and aware the general public or the target group of customers about a product a particular company is offering. The five energy drink companies – Speed, Tiger, Shark, Sting and Boost; all of them are between the introduction and growth stage of their product life cycle. At this stage they are only informing the customers about their product including their benefits. Basically all the companies are following the same way to promote their products and these are some of the most common media channels used by them for doing promotion – television, radio, newspapers, magazines and internet. Target Group: Energy drink companies mainly target the major districts of Bangladesh. The local companies, like – Speed, Tiger have distributed their products in rural areas as well to attract a huge number of customers. Energy drinks are mainly used by the people who are between the age group of 16-30, because they comparatively need more energy in every step of their lives. So all of the companies aim to target them and make the product according to their needs to deliver complete satisfaction through their energy drinks. Their product is priced quite cheaply, so that people of different classes can buy them. Most of the energy drinks are claimed to be halal while considering the religious and social issues of Bangladesh. Message and Effect of the advertisements: Message (Add Story) Each company make their advertisements differently even though their motive remains the same, which is to inform the customers about the product. Here are some of the advertisement stories of the five energy drink companies – 1. Speed In one of Speed’s advertisements, they have shown some guys were trying to get up on a bamboo for the whole day without getting tired. Their motto is ‘Heavy Energy’. In other words, they are using emotional appeal. In newspapers, they post a picture from the screen shot of their advertisements and use the same motto, so that people can get a clear brand knowledge. They do not have any brand ambassador for their product and they are constantly sending out the notion to their consumers how Speed can let them gain unlimited energy. 2. Shark There is an advertisement of Shark ‘20 takay Dhaka’, they showed a guy kicked another person from Chittagong and sent him to Dhaka after having Shark energy drink. They are using the motto ‘Real Energy, Real Power’. In their advertisements they use the rational appeal to attract customers. They have no brand ambassador, because they are not still going for intensive or aggressive promotion. 3. Sting -. In the television advertisement there was one young boy who went to a shooting spot with the hope of being a superstar. On that time one director was taking the shot of riding a motor cycle on that spot. Showing that the young boy moved forward to do that shot but the director did not allow him and insulted him that made the young boy very frustrated. After that the director was supposed to make that shot with the hero but unfortunately the hero failed to ride the motor cycle. Then the director asked the crowd who could do the shot. The first young boy showed up the courage and did that shot in a fantastic way. Everybody praised him a lot. They started celebrating the happiness together by dancing with sting energy drink. They were singing â€Å"Sting, Sting, Sting –Ullasher Shuru Ekhanei†. They are using rational appeal in their advertisements In the radio advertisement they play the same jingle. In newspaper the marketers use the picture of 250ml bottle of sting energy drink and write their jingle. They give the same ad in magazines too. In billboards and posters, the marketers add the picture in a big way that they use for newspaper and magazine. 4. Boost In one ad they have shown that a young guy, probably of age 20 was walking along the roadside, on the background rap music is going on (sign of young generation). The lyrics was talking about getting energy, confidence and success. The guy had his neck tattoed and wore a sunglass, showing the style of recent youth fashion. He went into a departmental store, went to the refrigerator, there were other drinks (e. g. red rooster and red bull); but he picked up BOOST, then he paid for it, got out, drank it and got himself recharged and the soundtrack was saying as if he could fly now. So this is one example of their rational advertisements. They have not used any brand ambassador yet for their product in Bangladesh, and these two advertisements are not even shown in Bangladeshi channels, they are only available in You tube. 5. Tiger The tiger energy drink has only one advertisement which is very popular. The brand ambassador Aiyub bachchu is the model for the advertisement. The advertisement showed that, when the popular singer Aiyub Bacchu attended a party, the audience requested him to sing a song. But at that time Aiyub Bacchu felt tired and bored. So he needed to recharge himself to get energy. So he took tiger and then sang a song with more power and energetic voice. In this way they try to attract customers by using the rational approach. They also do radio advertisement with the song of Aiyub Bacchu. In billboard advertisement they promote their motto ‘Recharge Yourself’. Effect – Different advertisements effect’s consumers differently. In general, all of these advertisements encourage people to buy their products. When people get satisfied with one product, they talk positive things about it, this way companies can enjoy the benefits of -word of mouth. Customers get to know through these advertisements how their everyday lives are going to be benefitted if they have these energy drinks. Personal selling: Personal selling is no longer considered as giving door to door service. It is a much wider concept nowadays. Companies not only their sell their product to the customers, they also build a loyal relationship with them. To keep on and for maintaining such a relation, companies must be ready to deal with different problems their customers have regarding their product. Unfortunately, none of these five energy drink companies do these in Bangladesh. They are more focused on advertising only. Sales promotion: In general, energy drink companies do not give any special discount on their product to boost their sales. Companies give incentives to their channel members to keep their product in the front line of their shelves, so that customers choose to have it by first looking at it. Sometimes they give energy drink on their own to the customers, because it make the customers feel the product is of high quality and that is why the salesman is offering it to them. Besides people generally do not look out for products those are kept behind the shelves, as it will be very time consuming. Advertisements do even contribute to generate the sales level sometimes. Though there are many other ways to promote sales, the energy drink companies mainly follow these two ways to carry out their sales promotion activity. Public Relation: To operate in this digital world it is very important for companies to maintain a good relation with both their internal and external publics, example – shareholders, customers. Some of the energy drink companies arranges concerts and sponsors television shows to interact closely with their customers. Example – last year Tiger organized one concert in Doha. Each of the five companies uses different and unique logos for their product. None of them are involved in doing conferences or giving press release. Buzz marketing is also a tool used by these companies to improve their relations with the general public and promote their product as well. Direct Marketing: Energy drink companies have their official websites, but their use is not very effective in Bangladesh. In huge shopping malls or in other areas, none of the energy drinks are available in vending machines. These products are not even sold by the companies directly over online or by using telephone. The only way they carry out direct marketing is through placing the advertisement videos in You Tube. The energy drink companies still lack good customer service, example – no service or facilities provided to give complaints or to share ideas about the product face to face with the customers. Promotion Mix Strategy: The demand for energy drink is not yet very high in Bangladesh. So all the manufacturers of these five energy drink companies use the push strategy. Manufacturers contract with wholesalers or retailers to pass down their products to their final consumers and also to encourage customers to buy the products. Sometimes manufacturers give incentives to the channel members to promote their product as well. Section 02: Marketing strategy for a new brand Figure: Spirit Energy Drink Figure: Logo of Spirit Energy Drink 2. 1: Introducing a new brand for energy drink After analyzing the present market condition of energy drinks in Bangladesh, we have come up with a new idea of introducing a new brand for energy drink. The name of our energy drink will be ‘SPIRIT Energy Drink’. This new energy drink will hopefully win the hearts of our target customers and please them with the new taste and value of our product that we going to offer. This energy drink will not only make people active physically, it will also motivate them spiritually to move ahead in their lives with confidence. The motto of our energy drink is ‘ignites you from inside’. With this motto we will give our customers the idea of how Spirit energy drink can add more liveliness in their lives. The name of our company is Spirit Beverage Limited and we are using ‘Spirit Energy Drink’ as our product’s logo. We have used picture of a guy inside the logo to show our customers what we are trying to say through the motto.roductinkurny is gy of Shark and make them more active. customers tend to buy more. 20202020202020202020202020202020. 2. 2: Design marketing strategy for the new brand i. Segmentation We will segment the market of Spirit energy drink on the basis of geography, demography, psychography and behavior. Geographic Segmentation: We are going to serve Spirit energy drink mainly for the people of Bangladesh. We will offer Spirit energy drink within the different territories of Bangladesh, like- Dhaka, Chittagong, Rajshahi, Barisal, Sylhet, Rangpur and Comilla. We will be selecting these territories because these are the biggest district of Bangladesh and most of the people live in these places. We will offer our product throughout every individual district, starting from the departmental stores to the small local and street stalls. We are offering Spirit both in urban and rural areas as well. Demographic Segmentation: Our market will be divided into groups based on variables such as age, gender, family life cycle, occupation and education. We are going to offer our product to both male and female who are between the ages of 15 to 30. We are especially offering the product to the university going students, corporate persons and athletes too. Our product will be for the people of all religion as it will be product is 100% halal. Psychographic Segmentation: Spirit energy drink is for the classes of people who fall under upper lowers, working class, middle class and upper middles. We are going to use personality variables to segment market like the gregarious and ambitious people. Behavioral Segmentation: We will be segmenting the market of Spirit energy drink on the basis of occasions and benefits. In summer it remains very hot in the middle of the day and for that time we will give a special offer to the customers called â€Å"Make Your Mind Cool†. People will get the same energy drink at a lower price during this time. We will give people other benefits like- we are not using preservatives in our product and our product will be convenient for the customers. ii. Targeting We will target our market within the segmentation. We are mainly targeting Dhaka because, it is the capital city of Bangladesh and highly populated and urbanized. Within Dhaka, we are targeting mostly the teenagers and adults who are the university students. Female students are going to be a part of it too, because the size and growth rate of this segment is high. As there is going to be other substitute products and tough competitors of Spirit energy drink in the market, we will offer our product directly by setting some temporary stores of Spirit energy drink inside the campus of some big and renowned universities. This will help us gain competitive advantage over our rivals and make people aware of our product. We will explain about the benefits about our product and give other information’s regarding our product. Making people believe that our product is different will be our main target. We will offer the same product at the same price to all our segmented customers. iii. Positioning Our product’s motto is â€Å"Ignites you from inside†, by this motto we are trying to make people believe that, Spirit energy drink encourages people to move on no matter whatever situation they face in their lives. Our product will always remove the frustration from both their body and soul. Spirit is alcohol free and will be made and designed attractively to attract our customers. In this way we will differentiate and create a unique image of our product in our consumer’s mind. iv. Marketing Mix (4p) ? Design and description of the product Customer Value and Brand Name: The first thing that we are considering about our product before we place it in the market is, it’s core benefit or core values. Customers always seek out for the benefit and the reason behind buying a product. They always value the experience they get after using it. In order to enter the market and establishing our brand successfully, we are going to provide the most refreshing and unique energy drink; which will spread liveliness and create more excitement in our customers life. They will not only buy one Spirit energy drink, they will buy one reason to hold on to being strong and face everything with more spirit in their lives. The brand name of our product is–‘Spirit Energy Drink’. We have named our product Spirit because it is easy to remember and meaningful at the same time. It will help the customers to distinguish and compare our product from others. Product Features and Design: Our energy bottle will be made in the shape of a space shuttle, with a combination of red and black color to give the igniting image. We will design our can just like other energy drink cans, which is made up of steel body with a cylindrical shape. In the middle of both the can and bottle, the name of our product will be written horizontally. The whole bottle and can will be black in color and the name written in pearl white. The shining stars above the name will give it a glowing and sparkling image. The guy standing in the middle of the name will exhibit the idea that Spirit energy drink has the ability to bring back everyone from darkness and add new value to their lives. Like all other energy drink, Spirit’s taste will be sweet and sour and the color of the drink will be red wine color. Packaging and Labeling: We plan to do very careful and colorful packaging to attract our target customers. The products are going to be well packaged and covered so that it stays protected in every condition. All the necessary information’s will be written on the body of bottle and can. Packaging will be done in two ways: 1. Plastic bottle – 300ml (Normal Packaging) 2. Steel Can – 270ml (Prestigious Packaging). Necessary warnings will be provided, like –it might be harmful for pregnant women and children. Manufacturing and Expiring dates will be written on the covers to warn the consumers about the product’s validity. Most importantly, the ingredients we are going to use are Carbonated water, Refined sugar, Sodium Citrate, Sodium Benzoate, Vitamin B3, Vitamin B6, Touraine, Citric Acid, Preservatives, Antioxidant, Ca-D-Pentothenate, Tartrazine, Mixed Fruit Flavor, Food Grade Flavor, Food Colors. There will be a Nutrition Fact chart too, describing the nutritious contents. Energy80 cal. Proteino gm Saturated fato gm Total fat o gm Sodium 20mg Sugar20 gm Carbohydrate 20. 5 gm Cholesterol0 gm Dietary Fibero gm Product Quality: Providing our customers with the best quality of energy drink is our prime purpose. We have made sure that our energy drink does not contain any harmful substances. It is alcohol free and free from every possible harmful effects. With the amazing taste and other attributes, we assure that our customers will be satisfied and buy Spirit more frequently. Design of pricing policy. Spirit is at the introduction stage of it’s product life cycle and we know the competition of energy drink companies is rising day by day in Bangladesh. We will use cost-plus pricing for Spirit energy drink, considering the present market and demand for energy drink. Though the demand of energy drink in our country is increasing with time, our product is still under pure competition, as there are already some other energy drinks in our local market. We will also consider competitors strategies and prices, how they set their prices and how much they charge from the intermediaries and consumers. We will focus on our target customers willingness and ability to pay for an energy drink. After considering the above issues we are charging 35tk for 270ml can and 30tk for 300ml bottle from our customers. For the 270ml can, our production is 30tk and after setting a standard markup to the cost we will be charging 32tk from our wholesalers, wholesalers will charge 33. 5tk to the retailers and finally the retailers are sell the drink at 35tk o the customers. For 300ml bottle, our production cost per bottle is 25tk and after setting a standard markup to the cost the wholesalers will buy each bottle at 27tk from the us. Wholesalers will charge 28. 5tk to the retailers and finally the retailers will sell it at 30tk each bottle to the customers. In short, we are charging the price for our product moderately to attract huge number of customers to buy it. It will help us to maintain a minimum level of profits for operating and keep running our business. Design of promotional activities – Advertising: For creating awareness and informing the consumers about our product we are going to advertise in more than one media. Our target is to do mass promotion to cover the maximum number of people in Bangladesh. Some of the major media channels through which we will be advertising are –television, radio, newspapers, online media and billboards. Television and internet are the most fastest and effective media channels to promote product, as most of the people spend their time on these. Newspapers though have high believability, do not last for a long period of time. Starting from the poor uneducated people of our society to the educated ones, everyone listens to radio. Therefore radio is also a very effective way to reach a huge number of customers at the same time. Add story While broadcasting on television, we will show a random university student is sitting on a chair and lost deep inside at the sheer thoughts of failing in exam. Then suddenly one of his friends will come to him and give him a Spirit energy drink. After having the energy drink the boy will jump off and scream energetically that he is not going to give up. He will get motivated and feel more energized while doing his studies. He will realize that he can study longer and give more concentration in his studies after drinking the Spirit energy drink. Then finally when the results come out, he will do the top result in his university. He will then become extremely happy and appreciate how Spirit energy drink has helped him to achieve this success in life. This way we will draw conclusion to the add. In newspapers and billboards we will use the same boy, who will be smiling and holding a Spirit energy drink; conveying the same message about our product’s motto. In radio channels we use the same jingle which we will show in television. As we are launching a new product in the market, we want to deliver the same message to our target customers. It will help them to get a clear and specific idea of our brand and what we intend to provide. Our advertisement is designed in both emotional and rational way. Emotionally we are showing the customer, they can move on in their lives with the help of Spirit and rationally we are giving them the idea that Spirit energy drink is good for their health, as it increases concentration and strength. Sales Promotion: Sales promotion is another major step we will carry out to encourage our customers to purchase our product more frequently. Sales promotion can be done in many ways – giving discounts, giving lotteries, pop-up adds, etc. We will give incentives to the people of our channels to keep our energy drink in front of the shelves in all the stores we are going to supply. Consumers buys the product that are kept in front side, because people have an idea that good quality things are always kept at the front and they will not want to search for other product as that would be time consuming. In times of special occasions like Eid, we will announce special discounts or extra offers. We will sell our products in front of colleges, universities and major shopping malls. Therefore we will be exposing our product more to the public and will gain higher chances of selling our product, thus making more profits. Besides, none of our competitors are doing these things, so we can gain competitive advantage over them as well. Personal selling: Energy drink is a convenience product, and it is quite impossible to reach the mass people by employing thousands of employees for selling our product. This process is very slow and expensive. So we will not be selling our products personally. We will employ experts in offices, who will deal with customer’s view’s and problems. This way we will personally provide/sell our service them beside selling our product. Public relations: At the very first stage we will arrange a press conference to inform everyone about our product. Press media is very crucial, as interacting with them positively will encourage them to promote our product more creatively and positively to our customers. This way we can also have competitive advantage over our competitors. We will organize different concerts and sponsor television shows, this will give us an open platform to interact more closely with our customers and build a strong relation with them. Direct marketing: We are planning to open a Facebook page along with our official website where we will encourage people to share their views about our product. This will help us to keep track of our customers reaction towards our product. None of the other energy drink companies are doing much of these activities, so it is an easier way to grab a huge number of customers and market share this way. Online marketing is therefore more efficient and fast way of direct marketing in terms of receiving our customers reactions, complaints and comments and adapting to those changes to keep surviving and growing. Promotion Mix strategy: We will use push strategy for Spirit energy drink. Contracting with different wholesalers and retailers will help us to provide adequate supplies in every convenient and targeted locations. We will give them incentives to promote and supply our product more frequently to the customers. Design of distribution activities- We will go for intensive distribution for Spirit energy drink, because we want to provide stock our product in as many outlets as possible. We want to ensure that our products must be available where and when consumers want them. As our product is new in the market, we will directly offer our product to the target customers. We will be offering our Spirit energy drink to the customers through wholesalers and retailers. Within our limited resources we cannot afford to have our own distribution facilities, like warehouses and showrooms and to hire sales people and support stuffs. We would rather contract with the wholesalers, so that they will take those extra costs and risks. Our target customers are the young university students, that is why we are contracting with retailers.

The value of the information you find on the internet Essay Example for Free

The value of the information you find on the internet Essay You are asking some very good questions. I often wondered the same before I learned about this subject. I would be glad to answer your questions and tell you what they all mean. First, you ask ‘How to assess the value of the information you find on the internet’? Well, there are many ways to assess the information you find on the internet. The first of many ways that I use to assess this information is to check the author information, meaning is the author trustworthy. To do this I often will see if the author has background in the subject that I am researching. I think that by doing this the information written by those who do not have any experience in the subject will not be on my radar. This also allows you to determine whether or not an author is authentic. Another important thing to check to see if information is reliable is to check the date, checking the date will help you determine if the information is up-to-date and if it has been updated recently. I think that by making sure the information is not outdated will help the credibility of this information. thing to check to see if information is reliable is to check the date, checking the date will help you determine if the information is up-to-date and if it has been updated recently. I think that by making sure the information is not outdated will help the credibility of this information. Another thing you would need to check is the website dependable. When I say ‘dependable’ I mean is the website easily edited by others who do not have any experience in the field? Has this information edited recently, and by whom? When using the internet you must be cautious of the information you read because some information could be false. You also ask about the ‘digital divide’ and what it means. The best way I can describe it is the divide between people who are up to date with all of the technological advances and those who are not. The people who are up-to-date with all of the technology advances have many ways to access all volumes of modern media though the internet. Whether it is though the use of a smart phone or the use of a computer or another type of technical device, whereas the people who do not have access to these types of media, people who rely on more traditional media such as, newspapers, books, and magazines would be on the opposite side of the digital divide. To bridge the digital divide all people would have to be able to access all versions of mass media. Meaning everyone would need to have to have equal  access to the internet. By that I mean that everyone should have access to the internet. Whether at home or otherwise would be keeping everyone in the technological world would definitely help Bridge the gap in the digital age.

Plant DCL Proteins

Plant DCL Proteins Abstract Dicer, a double-stranded RNA (dsRNA)-specific endoribonuclease, plays an essential role in triggering both transcriptional and post-transcriptional gene silencing in eukaryotes by cleaving dsRNAs or single-stranded RNAs bearing stem-loop structures such as microRNA precursor transcripts into 21- to 24-nt small RNAs. Unlike animals, plants have evolved to utilize at least four Dicer-like (DCL) proteins. Extensive genetic studies have revealed that each DCL protein participates in a specific gene silencing pathway, with some redundancy. However, a mechanistic understanding of how the specific action of each DCL protein is regulated in its respective pathway is still in its infancy due to the limited number of biochemical studies on plant DCL proteins. In this review, we summarize and discuss the biochemical properties of plant DCL proteins revealed by studies using highly purified recombinant proteins, crude extracts, and immunoprecipitates. With help from co-factor proteins and an ATP ase/DExH-box RNA-helicase domain, the microRNA-producing enzyme DCL1 recognizes bulges and terminal loop structures in its substrate transcripts to ensure accurate and efficient processing. DCL4 prefers long dsRNA substrates and requires the dsRNA-binding protein DRB4 for its activity. The short-dsRNA preference of DCL3 is well suited for short-RNA transcription and subsequent dsRNA formation by coupling between a plant-specific DNA-dependent RNA-polymerase IV and RNA-dependent RNA-polymerase 2 in the transcriptional gene silencing pathway. Inorganic phosphate also seems to play a role in differential regulation of DCL3 and DCL4 activities. Further development of biochemical approaches will be necessary for better understanding of how plant DCL proteins are fine-tuned in each small RNA biogenesis pathway under various physiological conditions. Introduction RNA silencing, also known as RNA interference (RNAi), is one of the fundamental molecular mechanisms conserved in most eukaryotes to regulate gene expression both transcriptionally and post-transcriptionally. In both situations, what triggers the RNA silencing pathway is a small RNA molecule, 21 to 24 nt in length, called small interfering RNA (siRNA) or microRNA (miRNA) depending on its origin and the downstream pathways involved. The class 3 endoribonuclease (RNase) III enzymes known as Dicer are responsible for producing siRNA from longer double-stranded RNAs (dsRNAs) and miRNA from single-stranded RNAs with internal stem-loop structures   by a dsRNA-specific endoribonuclease. Therefore, the activity and regulation of Dicer-family proteins in a cell are vital to many biological processes requiring flexible adjustments at the level of gene expression, such as development, organogenesis, the circadian rhythm, biotic and abiotic stress responses, and defense against viruses an d transposons. Biochemical characterization of Dicers in animals The Dicer family is a unique class of RNase III enzymes due to the presence of an ATPase/DExD/H-box helicase domain at the N-terminus, a Piwi/Argonaute/Zwille (PAZ) domain in the middle and dual RNase III domains followed by one or two dsRNA-binding domains in the C-terminal half (exception: Giardia intestinalis) (Figure 1) (Bernstein et al. 2001). In general, the helicase domain serves as a protein-protein interaction surface recruiting co-factor regulatory proteins (Lee et al. 2006; Ma et al. 2008; Ye et al. 2007). It also utilizes ATP hydrolysis to achieve processive cleavage of the long dsRNA substrate (Cenik et al. 2011; Welker et al. 2010). The PAZ domain contains a conserved pocket for recognizing the terminus of the dsRNA substrate, and the distance between PAZ and the RNase III catalytic center determines the product sizes (MacRae et al. 2007; MacRae et al. 2006). Each of the two RNase III domains cuts one of the dsRNA strands, leaving a characteristic 2-nt overhang at 3-end of the product (Elbashir et al. 2001; Takeshita et al. 2007; Zhang et al. 2004). The C-terminal dsRNA-binding domains (dsRBDs) serve as a protein-protein interaction interface and nuclear localization signals, in addition to having dsRNA-binding function (Doyle et al. 2013; Hiraguri et al. 2005; Wostenberg et al. 2012). The specific functionality of each domain differs depending on the Dicer protein. Since the first demonstration of in vitro small RNA-producing activity of Dicer in the fruit fly Drosophila melanogaster (Bernstein et al. 2001), its biochemical properties and regulatory machinery have been extensively studied in humans, D. melanogaster and Caenorhabditis elegans. In humans, there is only one Dicer-family protein (hDicer), which cleaves short-hairpin pre-miRNAs produced by Drosha and dsRNA substrates into 20- to 22-nt small RNAs in an ATP-independent manner (Myers et al. 2003; Provost et al. 2002; Zhang et al. 2002). The cleavage activity requires a divalent metal cation such as Mg2+, Co2+ or Mn2+, and recognizes mainly the 5-end of the substrate to dictate the product length (Park et al. 2011). This 5-counting rule is reliant on the conserved 3-pocket motif within the PAZ domain and the 5-pocket motif, which is less conserved in Dicers of other eukaryotes. The binding of Dicer to a dsRNA substrate and its cleavage are uncoupled, because Dicer can bind to dsRNA with out Mg2+ or under low temperature (Provost et al. 2002; Zhang et al. 2002). The helicase domain of hDicer has an autoinhibitory function (Ma et al. 2008). In line with this, the activity of recombinant full-length hDicer protein can be improved under limited proteolytic conditions (Zhang et al. 2002). hDicer is responsible for both siRNA and miRNA production, and co-factor dsRNA-binding proteins TRBP and PACT dictate hDicer function in the two distinct small RNA production pathways (Chendrimada et al. 2005; Haase et al. 2005; Kok et al. 2007; Lee et al. 2013; Lee et al. 2006). In particular, the hDicer complex containing PACT disfavors siRNA precursor dsRNA and shows different cleavage patterns on the same pre-miRNA substrate than the hDicer-TRBP complex (Lee et al. 2013). The interaction with TRBP occurs through the hDicer helicase domain, and stimulates the hDicers catalytic activity. (Ma et al. 2008). Similarly, it has been reported that the C. elegans Dcr-1 interacts with a dsRNA-binding protein RDE-4 which enhances the Dicer activity toward long dsRNA substrates in siRNA production, while RDE-4 is apparently dispensable in miRNA production pathway (Parker et al. 2006; Parker et al. 2008; Tabara et al. 2002). D. melanogaster has two Dicer proteins, Dcr-1 and Dcr-2, which produce miRNA and siRNA, respectively (Lee et al. 2004; Miyoshi et al. 2010). Dcr-1 alone can process dsRNA into siRNA in vitro, but its interaction with the dsRNA-binding protein Loquacious isoform PB (Loqs-PB) confers pre-miRNA substrate specificity to the Dcr-1-Loqs complex by suppressing cleavage of long perfect dsRNAs and enhancing pre-miRNA processing activity (Saito et al. 2005; Zhou et al. 2009). Dcr-2 interacts with Loqs isoform PD and another dsRNA-binding protein, R2D2, in the siRNA production pathway (Liu et al. 2003; Liu et al. 2006; Miyoshi et al. 2010; Zhou et al. 2009). Dcr-2 alone is also capable of cleaving a pre-miRNA precursor in an ATP-independent manner, but R2D2 significantly suppresses Dcr-2 activity toward pre-miRNA, whereas Loqs-PD enhances the cleavage activity of Dcr-2 toward long perfect dsRNA precursors by boosting its affinity to the substrate (Cenik et al. 2011; Miyoshi et al. 2010). The pr ocessive processing of long dsRNA substrates by Dcr-2 depends on ATP hydrolysis by its ATPase/helicase domain, implying that one of the functions of the helicase domain is to allow Dcr-2 to produce multiple siRNAs from a single long dsRNA molecule before it dissociates from the substrate (Cenik et al. 2011). Such differential regulation of Dicer activity through specific interaction with co-factor dsRNA-binding proteins in distinct pathways is commonly found in most of the systems studied, including plants. DCL proteins in plants Plant genomes contain at least four distinct classes of DCL family proteins (DCL1-4). Like their animal counterparts, each class of DCL has evolved to participate in its primary pathway (Figure 2), but the three siRNA-producing DCLs (DCL2-4) function redundantly as well, because defects in one class of DCL can be compensated for by other classes in some cases (Gasciolli et al. 2005; Mukherjee et al. 2013; Xie et al. 2004). Because DCL1 is the only Dicer protein that produces most 21-nt miRNAs (Kurihara and Watanabe 2004; Reinhart et al. 2002), knockout mutants of DCL1 are embryonic lethal (Schauer et al. 2002). DCL4 is the major producer of 21-nt antiviral siRNA and endogenous siRNAs such as trans-acting siRNA and phased siRNAs (phasiRNA) (Bouche et al. 2006; Gasciolli et al. 2005; Mukherjee et al. 2013; Qu et al. 2008; Xie et al. 2005; Yoshikawa et al. 2005). DCL2 can compensate for the loss of DCL4 (Bouche et al. 2006; Gasciolli et al. 2005; Parent et al. 2015), although its major function remains unclear. DCL3 mainly produces 24-nt repeat-associated siRNAs derived from transposons and DNA repetitive elements, and participates in transcriptional gene silencing (TGS) through RNA-dependent DNA methylation, suppressing proliferation of these elements (Henderson et al. 2006; Pontes et al. 2006; Xie et al. 2004). In addition to the four classes of DCLs, monocots have another distinct class of Dicer, DCL5 (also known as DCL3b) (Margis et al. 2006). DCL5 is specifically expressed in developing panicles and is responsible for 24-nt reproductive phasiRNAs, although the biological significance of a reproductive-organ-specific 24-nt phasiRNA pathway mediated by this specific Dicer remains to be elucidated (Borges and Martienssen 2015; Fei et al. 2013; Kapoor et al. 2008; Song et al. 2012). This pathway might be analogous to the Dicer-independent PIWI-interacting RNA (piRNA) pathway in vertebrates, which suppresses transposons and other genes specifically in germlines (H irakata and Siomi 2016). Both forward and reverse genetics and physiological studies have successfully dissected the major RNA silencing pathways and allowed identification of the function of DCL genes in each pathway in plants. However, investigations on the molecular and enzymatic characteristics underlying the functional diversification and specificity of the DCL proteins are still in their infancy. Detection of DCL activity in crude extracts of various plants Biochemical characterization of plant Dicer activity was first demonstrated in wheat germ extract (monocot) and cauliflower extract (dicot), which contain multiple DCL activities producing ~21 nt and ~24 nt small RNAs with 2-nt 3-overhangs in the double-stranded form (Tang et al. 2003). These activities are weaker in the absence of ATP, consistent with characteristics of Dicer family proteins from Drosophila and C. elegans. Long dsRNA competitors effectively suppress both activities in wheat germ extract. The 24-nt small RNA producing activity was inhibited by 25-nt synthetic siRNA duplexes, whereas 21-nt small RNA production was unaffected by 21-nt synthetic siRNA duplex competitors, suggesting that two different enzymes with active sites that have distinct size-dependent binding properties are in the wheat germ extract (Tang et al. 2003). A recent study on wheat germ extract characterized these activities in further detail, revealing (1) that the 21-nt activity could be found in a much larger (~950 kDa) complex than the 24-nt activity, which had maximum activity in an approximately 450 kDa complex; and (2) the biochemical properties associated with the activities, such as divalent cation and NTP requirements, optimum NaCl concentration, temperature, and pH, and substrate length dependence (Shivaprasad et al. 2015). The identities of the DCL enzymes responsible for these activities in the wheat germ extract remain to be identified. A better understanding of the biochemical characteristics of individual plant Dicer proteins has come from the model plant Arabidopsis thaliana, which has four DCL proteins: DCL1, DCL2, DCL3 and DCL4 (summarized in Table 1). The first in vitro DCL activity in A. thaliana was demonstrated using a suspension cell lysate, a crude extract of inflorescence tissue, and an immunoaffinity-purified protein complex (Qi et al. 2005). Similar to the previous study using wheat germ extract or cauliflower, extracts from both Arabidopsis cultured cells and inflorescence tissue contained DCL dsRNA-cleaving activity producing 21- and 24-nt small RNAs from 400-bp dsRNA (Qi et al. 2005). The 21-nt producing activity and 24-nt producing activity were found in >660 kDa and ~400 kDa fractions, respectively, suggesting that these Dicers reside in protein complexes composed of multiple co-factors (Qi et al. 2005). In agreement with previous genetic studies showing CARPEL FACTORY/DCL1 is responsible for 21-nt miRNA production in vivo (Kurihara and Watanabe 2004; Reinhart et al. 2002), the 21-nt small RNA producing activity was DCL1 immunoaffinity-purified from inflorescence-derived crude extract by an anti-DCL1 antibody (Qi et al. 2005). The 24-nt activity was associated with anti-DCL3 antibody immunoprecipitate, and the activity was abolished when purified from a dcl3-1 mutant, showing that DCL3 is responsible for the 24-nt activity in Arabidopsis inflorescence extract. The immunoaffinity-purified DCL1 activity required ATP, whereas the activity of the DCL3 immunoprecipitate was ATP-independent (Qi et al. 2005). Interestingly, the dcl1-7 mutation did not abolish the 21-nt small RNA producing activity in the extract or immunoprecipitates, implying that the substitution (P415S) in its N-terminal helicase domain did not alter the enzymes catalytic activity itself (Qi et al. 2005); this study also found that the activity of DCL4 responsible for formation of 21-nt siRNA was present in the inflorescence extract. The presence of DCL4 activity in an Arabidopsis crude extract was demonstrated in later studies using 2-week-old seedlings as the starting material (Fukudome et al. 2011; Nagano et al. 2014), and will be discussed later in this review. In-depth biochemical characterization of DCL1, a microRNA-producing enzyme in plants  DCL1 activity requires DRB1/HYL1 and SERRATE for accurate processing of the miRNA precursor Both in wheat germ and Arabidopsis extracts, DCL activities are associated with size fractions larger than DCL monomeric form, implying that these DCLs form functional protein complexes composed of multiple co-factors in vivo. As summarized in an earlier section, such interactions between a Dicer and a co-factor protein are commonly found in mammals, nematodes and insects. One of the most characterized classes of co-factor proteins is a dsRNA-binding protein (dsRBP) harboring multiple dsRNA-binding domains or motifs. The A. thaliana genome encodes five dsRNA-binding (DRB) family proteins: DRB1/HYL1, DRB2 DRB3, DRB4, and DRB5. Multiple genetic and biochemical studies have demonstrated two specific interactions between DCLs and DRBs in A. thaliana: DCL1-DRB1/HYL1 and DCL4-DRB4 (Han et al. 2004; Hiraguri et al. 2005; Kurihara et al. 2006; Nakazawa et al. 2007). Arabidopsis DCL1, DRB1/HYL1, and another co-factor, SERRATE (SE), constitute an essential microRNA production pathway in vivo (Han et al. 2004; Lobbes et al. 2006). Unlike animals, which utilize two distinct RNase III enzymes, Drosha and Dicer, for the first and second cleavage of microRNA precursors, plants do not employ Drosha. Therefore, the DCL1-complex is responsible for the processing of both primary and precursor miRNA substrates. The detailed molecular machinery of the dual miRNA processing mediated by DCL1 and the co-factor proteins have been extensively studied biochemically using highly purified recombinant proteins produced in heterologous systems (summarized in Figure 3). One of the systems utilizes baculovirus-mediated recombinant protein production in Sf21 insect cells, followed by two-step affinity purification (Dong et al. 2008). The highly purified recombinant DCL1 protein alone could process a 94-bp dsRNA substrate with a 2-nt 3-overhang into 21-nt small RNA in an ATP/Mg2+ dependent manner. The optimum NaCl concentration for the activity was 25-50 mM, and a NaCl concentration higher than 100 mM severely impaired the activity (Dong et al. 2008). While the recombinant DCL1 protein alone could produce 21-nt small RNA from both primary and precursor miRNA (pri-/pre-miR167b) substrates in vitro, DRB1/HYL1 and SE recombinant proteins co-incubated in the same reaction mixtures significantly increased both yield and accuracy of the processing (Dong et al. 2008). Without these co-factors, more than 80% of 21-nt small RNA products from the DCL1-alone reaction were due to incorrect processing from the end of the primary miRNA substrate, whereas the processing mediated by the DCL1-DRB1/HYL1-SE complex produced accurate 21-nt products with a sequence identical to miR167b/miR167b*, amounting for up to 81% of the products (Dong et al. 2008). This demonstrated that accurate processing of miRNA precursors by DCL1 requires the co-factors DRB1/HYL1 and SE. Cons istent with a previous study, the interaction between DCL1-DRB1/HYL1 through the second dsRNA-binding motif of DCL1 is important for the precise processing of pri-miRNA in A. thaliana (Dong et al. 2008; Kurihara et al. 2006). Also, using highly purified recombinant proteins and surface plasmon resonance analysis, it has been suggested that DCL1 changes its structural conformation when it binds RNA and exposes more binding sites for SE (Iwata et al. 2013). Binding to substrate dsRNA or miRNA precursors might be an important regulatory step for DCL1 dicing activity, as its dsRNA-binding domains exhibit the strongest binding to dsRNA among the four Arabidopsis DCLs (Hiraguri et al. 2005). ATPase/DExH-box RNA-helicase domain of DCL1 suppresses its dicing activity, confers ATP dependence, and influences processing accuracy In addition to its RNase III and dsRNA-binding domains, the helicase domain of DCL1 plays a significant role in regulating its dicing activity. Two independent forward genetic studies have identified two dcl1 mutant alleles, dcl1-13 (E395K) and dcl1-20 (R363K), as hyl1 suppressors, and the amino acid substitutions of both alleles occur within the ATPase/DExH-box RNA-helicase domain. These dcl1 mutations partially rescue the accumulation of some miRNAs in a hyl1-2 mutant (Liu et al. 2012; Tagami et al. 2009), and dcl1-13 was at least partially able to restore the phenotypic defects of hyl1-2 such as a reduced number of rosette leaves and a   leaf shape (Tagami et al. 2009). Highly purified recombinant DCL1-20 protein exhibited enhanced catalytic activity (Kcat/Km) toward pri-miRNA156a compared to wild-type DCL1 (Liu et al. 2012). Similarly, the helicase domain-deleted DCL1 recombinant protein (DCL1à ¢Ã‹â€ Ã¢â‚¬  Helicase) showed higher processing activity in vitro and was no l onger dependent on ATP for its activity toward pri-miRNA156a (Liu et al. 2012), suggesting that the helicase domain of DCL1 might have an autoinhibitory function like that of human Dicer (Ma et al. 2008; Provost et al. 2002). The in vivo miRNA processing imprecision in hyl1-2, however, was not restored by a dcl1-20 mutation, implying that the partial recovery of the hyl1-2 mutant, including miRNA accumulation, was due to the enhanced catalytic activity resulting from the substitution in the helicase domain (Liu et al. 2012). Interestingly, the effect and magnitude of DRB1/HYL1 and DCL1 helicase domain seem to vary among miRNA precursors. For example, the in vivo processing accuracy of miR156a is much more severely affected by hyl1-2 mutation than miR166b is (Liu et al. 2012). pri-miR156a is processed from the loop-proximal site to the loop-distal base in vitro (Liu et al. 2012), which is considered unusual for plant miRNAs (Addo-Quaye et al. 2009; Mateos et al. 2010). Accurate processing of pri-miRNA166b by native DCL1 is largely dependent on the presence of ATP, and processing by DCL1à ¢Ã‹â€ Ã¢â‚¬  Helicase is less accurate than that of native DCL1 (Liu et al. 2012). In contrast to miR156a, the processi ng precision of which is markedly affected by hyl1-2, that of miR166b was much more impaired by dcl1-20 mutation than hyl1-2 (Liu et al. 2012). Also, the effect of the other helicase mutant allele, dcl1-13, on miRNA production was shown to depend on the presence or absence of DRB1/HYL1 in vivo (Tagami et al. 2009). These observations indicate that efficient processing of different miRNA precursors by DCL1 have different reliance upon DRB1/HYL1 and DCL1helicase domain that potentially depends on structural determinants of the miRNA precursors. Structural determinants for efficient and accurate processing of miRNA precursors by DCL1 Primary transcripts of miRNA (pri-miRNA) have a characteristic secondary structure: a loop-distal stem (lower stem), a miRNA duplex, a loop-proximal stem (upper stem) and a terminal loop (Figure 3). Typical miRNA maturation from these precursors requires at least two cleavages occurring at the lower and upper stems. In animals, the single-stranded base region of the loop-distal stem is recognized by the dsRNA-binding protein DGCR8, which guides the processing center of Drosha to the correct position, which is 11 nt from the base of the stem (Han et al. 2006). However, this distance-from-base rule is not sufficient for plants because the length of the loop-distal stem of plant pri-miRNAs is highly variable (Song et al. 2010). Several structural features of pri-miRNAs that influence the activity, binding position and directionality of the processing by DCL1 have been elucidated genetically and biochemically (Figure 3a, b). One structural determinant lies within the loop-distal stem of pri-miRNA. For the first cleavage at the loop-distal stem, bulges and unpaired regions play a major role in the efficiency of miRNA processing. Mutant pri-miRNAs with closed bulges were processed at the correct position, but resulted in the accumulation of unprocessed pre-miRNAs in vivo, indicating that the rate of subsequent processing at the loop-proximal stem was impaired (Song et al. 2010). In pri-miR171a, which has a long loop-distal stem, the first cleavage position was determined by the distance from a relatively unstructured region instead of the base of the stem; the conserved distance from an unstructured region of the lower stem important for miRNA processing was found to be approximately 15 nt (Figure 3a) (Mateos et al. 2010; Song et al. 2010; Werner et al. 2010). The 15-nt rule was essentially reproduced in an in vitro miRNA processing system using highly purified DCL1-DRB1/HYL1-SE recombinant proteins and an artificial pri-miRNA substrate bearing another unstructured region in the elongated lower stem. In addition to the canonical processing, another type of processing occurred at 15 nt from the artificially introduced unstructured region, validating the functionality of the 15-nt rule (Song et al. 2010). The importance of bulges and unpaired regions in the lower stem for processing by DCL1 might explain why some miRNAs with a near-perfect matched stem seem to be DCL4-dependent, rather than DCL1-dependent (Rajagopalan et al. 2006; Song et al. 2010). On the loop-proximal and terminal loop side, a branched terminal loop (BTL) or a large terminal loop was found to be an essential structural factor that may alter directionality of processing by DCL1 and the resultant miRNA-accumulation (Figure 3b). BTL induces abortive processing of pri-miR166c both in vivo and in vitro (Zhu et al. 2013), meaning the first cleavage of the pri-miRNA occurs in the loop-proximal stem as opposed to the normal productive processing beginning in the loop-distal stem. The molecular basis of this bidirectional processing by DCL1 was further investigated using an in vitro system that reconstitutes the DCL1-processing machinery. For this purpose, DCL1, DRB1/HYL1 and SE harboring Agrobacterium tumefaciens were co-infiltrated to Nicotiana benthamiana leaves, and the transiently expressed DCL1-DRB1/HYL1-SE complex was immunoaffinity-purified two days after infiltration (Zhu et al. 2013). The reconstituted DCL1 complex cleaves the substrate pri-miRNA 16-17 nt fro m the unpaired region of the lower stem, supporting previous studies (Mateos et al. 2010; Song et al. 2010; Werner et al. 2010). By disrupting one of the two RNase III domains of DCL1 alternately and using 5- or 3-end labeled pri-miR166c substrates, the bidirectional nature of both productive and abortive processing was demonstrated (Zhu et al. 2013; Figure 3b). The helicase domain of DCL1 fine-tunes the position of both productive and abortive processing by DCL1 in an ATP-dependent manner (Zhu et al. 2013). DCL1à ¢Ã‹â€ Ã¢â‚¬  Helicase complex could not abortively process a substrate with BTL. Also, wild-type DCL1 required ATP for abortive processing, but not productive processing, indicating that the ATPase-driven helicase activity is necessary in abortive processing to unwind the structured BTL (Zhu et al. 2013; Figure 3b). In productive processing, the effect of helicase deletion and ATP depletion depend on the distance between the processing site and the bulge in the lower stem. Many potential byproducts of the abortive processing of pri-miRNA precursors with BTL can be found in publically available high-throughput small RNA sequencing data from both Arabidopsis and rice, implying that both substrate structure and the functionality of the ATPase/helicase domain of DCL1 are conserved mechanisms to regulate miRNA biogenesis in higher plan ts (Zhu et al. 2013). Dissecting distinct characteristics of DCL3 and DCL4 activities DCL4 activity requires the dsRNA-binding protein DRB4 In A. thaliana, DCL2, DCL3 and DCL4 are responsible for producing various siRNAs 21-24 nt in length. The dsRNA-cleaving activities of DCL3 and DCL4 can be detected in crude extracts prepared from 2-week-old seedlings (Fukudome et al. 2011). Extracts from wild-type seedlings cleave 500-bp dsRNA substrates into 21-nt and 24-nt small RNAs. In this system, the 21-nt and 24-nt small RNA-producing activities can be attributed to DCL4 and DCL3 respectively, because the dsRNA-cleaving activity of the corresponding size was abolished in each of the single mutants (Fukudome et al. 2011). Also, a mutation in the dsRNA-binding protein DRB4, which interacts with DCL4 (Hiraguri et al. 2005; Nakazawa et al. 2007), abolished DCL4 activity in seedling extracts. The DCL4 activity could be further purified by immunoprecipitation with anti-DCL4 or anti-DRB4 antibodies. The immunoaffinity-purified DCL4 requires Mg2+ and ATP for its activity, and is inhibited by >200 mM NaCl. This property is similar to t hat of recombinant DCL1 protein (Dong et al. 2008). The DCL4 complex immunoprecipitated from the drb4-1 mutant did not show dsRNA-cleaving activity, but the addition of recombinant DRB4 protein to the complex restored the 21-nt producing activity in vitro, showing that DRB4 functions as an essential co-factor for the dsRNA-cleaving activity of DCL4 (Figure 4b). In this system, mutant DRB4 proteins harboring substitutions in the conserved amino acid residues that form a hydrogen bond with the phosphodiester backbone of dsRNA at the dsRNA-binding site (H32A in the first dsRBD and K133A in the second dsRBD of DRB4) lost their ability to interact with dsRNA and DCL4, and did not restore DCL4 activity. The second substitution (K133A) alone impaired its interaction with the C-terminal half of DCL4 containing two RNase III domains and two dsRBDs in a GST pull-down assay using recombinant proteins, but was not sufficient to block restoration of DCL4 activity when added to DCL4 immunopurified from a drb4-1 mutant extract. There might be an add itional interaction surface between DCL4 and DRB4 involving dsRBD1 of DRB4 and the N-terminal half of DCL4, which contains an ATPase/DExH-box RNA-helicase domain and an RNA-binding domain (formerly known as domain of unknown function DUF283; Figure 1), as their specific interaction was reported in vitro (Qin et al. 2010). Short dsRNA preference of DCL3 activity orchestrates 24-nt siRNA biogenesis in TGS pathway Crude extracts from 2-week-old seedlings have also been used to characterize substrate specificity of DCL3 and DCL4. Consistent with the long dsRNA preference of Drosophila Dcr1 (Bernstein et al. 2001), DCL4 preferentially cleaves longer dsRNA substrates, and is less efficient in producing 21-nt siRNAs when the substrate is shorter than 50 nt (Nagano et al. 2014). On the other hand, DCL3 activity, producing 24-nt siRNAs, favors shorter substrates such as 30 nt and 37 nt dsRNA with a 1-nt or 2-nt 3-overhang (Nagano et al. 2014). It also favors substrate dsRNA with 5-adenosine or uridine. The 24-nt small RNA produced by DCL3 has a 2-nt 3-overhang, and the cleavage follows the 5-counting rule proposed for human Dicer (Park et al. 2011). DCL3 is not reliant on ATP hydrolysis for activity, as it can still process the short dsRNA substrate in the presence of a non-hydrolyzable ATP analog, adenosine 5-O-(3-thio)triphosphate (Nagano et al. 2014). Unlike DCL4, which targets long dsRNAs such a s RDR6-dependent TAS dsRNAs or exogenous viral dsRNAs in vivo (Bouche et al. 2006; Dunoyer et al. 2005; Qu et al. 2008; Yoshikawa et al. 2005), DCL3 may not need to perform a processive cleavage, which requires ATP hydrolysis, because the length of its targets allows only a single cut (Figure 4a). The DCL3 preference for short dsRNA substrate is consistent with the one precursor, one siRNA model for RNA polymerase IV (Pol IV)-dependent 24-nt siRNA biogenesis (Blevins et al. 2015; Zhai et al. 2015). In this model, a remarkably short (30- to 40-nt) transcript with 5-adenosine is produced by Pol IV and is simultaneously converted into double-stranded form by an RNA-dependent RNA polymerase, RDR2. The short dsRNA substrate is processed into 24-nt siRNA preferentially by DCL3 due to its length specificity, facilitating the subsequent RNA-directed DNA methylation process (Blevins et al. 2015; Zhai et al. 2015). The transcription of short RNAs by Pol IV, and the length and 5-adenosine substrate preference of DCL3 might be essential mechanisms to prevent other DCLs from processing specific dsRNA substrate needed for the TGS pathway. Such coupling of RDR-Dicer-RNAi is also known in fission yeast, where a Dicer physically interacts with an RNA-dependent RNA polymerase to form coupled ma chinery that drives siRNA-mediated TGS (Colmenares et al. 2007). In addition, DCL3 can participate in 24-nt siRNA production from longer transcripts with aid from another RNase III enzyme, RNase III-like 2 (RTL2). As a class II RNase III enzyme, RTL2 possesses one RNase III domain and two dsRBDs, and is involved in rRNA maturation [in vivo is implied]in A. thaliana (Comella et al. 2008). Recombinant RTL2 protein can cleave long dsRNA substrates into 25 bp or longer dsRNA in vitro (Kiyota et al. 2011). Recently, it has been shown that RTL2 processes a subset of Pol IV-dependent dsRNA into shorter intermediates, which are preferable for DCL3 activity in vivo (Elvira-Matelot et al. 2016). Although no direct interaction has been reported, RTL2 and DCL3 can be considered other examples of coordinated action of a dsRBD-containing protein and a Dicer in plants. DCL3 is also reported to physically interact with the dsRNA-binding protein DRB3 in the antiviral RNA-directed DNA methylation pathway (Raja et al. 2014). The function of DRB3 in DCL3 activity rem ains elusive. Inorganic phosphate, NaCl and KCl differentially regulate DCL3 and DCL4 activities In the same assay system using crude extracts, inorganic phosphate at a physiological concentration promotes DCL3 activity but suppresses DCL4 activity toward 50-nt dsRNA substrates (Nagano et al. 2014). The differential effe