Resume,letter,executive summary Essay Example | Topics and Well Written Essays - 1000 words

Resume,letter,executive summary - Essay Example I graduated in year 2009 with a degree in Electrical Engineering, minor in telecommunications applications. I took the state exam for Electrical Engineer Licensure in April of 2010 and passed with the grade of 93.8%. Although I do not have any substantial experience as a licensed Engineer just yet, I have had exposure working for an Engineering firm in 2009, whereby as was tasked to coordinate directly with the resident Electrical Engineers and to provide the necessary assistance in their operations. Some of the projects I was involved in are on semiconductors and vector works. Though limited as my hands on exposure could be, I believe that I was able to gain a considerable amount of knowledge to equip me as I take on my first real job after graduating and obtaining my license. The skills and expertise of an Electrical Engineer are required in construction projects, particularly on the assembly of electrical and electronic systems needed to mobilize the construction team, and post-project to provide the structure the needed electrical system to function according to its design. The person to hold an entry level Electrical Engineer position is not really expected to have intermediate experience on circuit breakers, transformers, induction motors, and other equipment. However, he must have basic knowledge on the principles and basic know hows. He must be able to discuss the principles and apply the concepts as he is prepared to go hands on later on after a period of immersion. Electrical Engineers in an entry level position are expected to assist in the design, implementation, maintenance and improvement of electrical instruments, facilities, products and equipment for various purposes, including commercial, domestic, and industrial purposes. They are also expected to be knowledgeable on the operations of computer-assisted engineering and design software to perform tasks needed to complete a project. As these personnel are still starting up their

The Code of Ethics for Nurses Essay Example for Free

The Code of Ethics for Nurses Essay This paper discusses the code of ethics of nurses. The nurses are sometimes found in very challenging situations that call for proper decision making. The action taken should not be against this code of ethics. Nurses are guided by a set of laws that are meant to assist them in their duties to offer medical services and at the same time satisfy their patient’s requirements. The code of ethics for the nurses rejects any attempt to perform activities that are contrary to the patient’s rights. These include the right to life, to self esteem and the right to respectful treatment (International Council of Nurses, 2006). In the case study situation the surgeon made a wrong recommendation for the patient. Mr. B was wrongfully scheduled for a tumor resection procedure yet the most appropriate course of treatment for him was a palliative treatment or limited intervention. Subjecting the patient to a tumor resection was an act that subjected the aged patient to a higher risk of postoperative recovery (Strang, Strang, Hultborn, Arner, 2004). With the problem of bed space the patient deserved to be treated equally with other patients despite his age, the health condition and his limited chances of survival. The human rights stipulate clearly that the life of one individual is equally important as the other and therefore should not be undermined (Tschudin, 2003). The nurses should have acted by the principle of the first patient to be admitted the first to be served and in that situation the other patients younger than Mr. B should have been directed to other facilities. By all ethical standards, nurses should always value the lives of their patients regardless of the situation since all the lives are precious and none can be substituted for another one.

Effects Of Rising Fuel Prices Economics Essay

Effects Of Rising Fuel Prices Economics Essay This research project is being written because currently the UK is experiencing a recession, which has escalated the price of fuel and is affecting the supermarkets. The research will examine the impact of fuel prices on different supermarkets and their approaches taken to boost sales and improve competitiveness. The intended reason for presenting this research project is to show how supermarkets can become more efficient and supportive. In order to answer the research question the following criteria will be addressed: Formulate a research specification Implement the research project within agreed procedures and to specification Evaluation of research outcomes Present the research outcomes The fuel prices will be compared and contrasted for the supermarkets during the past 5 years between 2008 2012. It will then make recommendations based on the findings of the research and analysis. This report is to be completed by 22nd April 2013. Methodology This research which is being provided has made use of different resources; lecture notes, textbooks, newspapers, discussions with family members and internet websites for the different organisations mentioned e.g. Asda, Morrisons, Sainsburys and Tesco In order to complement the secondary data derived from research studies and published material, telephone interviews with several supermarkets will be conducted. The interview will include questions for the supermarkets both, as a consumer and a supplier of fuel directly to its customers. The research objectives looking to be achieved by the end of this report are Using available secondary data, to establish the pattern and trends of the impact of fuel prices on supermarkets in the last 5 years Collecting primary data to obtain first-hand account from company personnel, about their decisions and adopted strategies to manage the impact of rising prices, both as a consumer and supplier of fuel To make a set of recommendations on fuel incentive programme based on core product offerings This study was conducted as part of a HND in Business unit which involves conducting research and then presenting this in the form of a research project which takes into account all the research gathered. The main purpose of this unit is to conduct research on a topic or any current issues. It was decided to conduct my research based around How the increase in the cost of fuel is affecting supermarkets in 2008 compared to 2012. In order to help obtain individual and business views on how the price increase is affecting them, this will involve the gathering of information from 1:1 telephone calls and face-to-face interviews with supermarkets. The completed questionnaires will help make an overall analysis of the data at the end, in order to produce an evaluative report based on the findings. With the research provided, this helped me look at gaining more insight into how different supermarkets are operating in the current recession and what changes would be made in order to cope with the increase in fuel prices. Therefore this led to the setting of the aim, about exploring further into how supermarkets are operating by the fuel price increase in terms of distribution and sales. With the production of this research project, the supermarkets will benefit the most alongside the oil companies and consumers. However, the petrol stations will also need information in order to succeed. The need for the report is to highlight the price variations and how supermarkets adapt to change. Methodology Chapter Currently the global economy is experiencing a recession. As a result of this, the economy and individuals are facing problems like a rise in unemployment levels which means it is becoming harder to get a job, fuel prices increasing and a decline in the overall market level. In conclusion the recession means that the business sector is hit the largest alongside consumers. When doing the project there will be various steps involved. Firstly this will establish the goal of the overall chosen research topic How the increase in the cost of fuel is affecting supermarkets, and then determine my sample in regards to views on how and why the fuel prices are affecting the supermarkets. The next step would be the creation of the questionnaire which is to be distributed amongst businesses and those being interviewed. The research collected from the questionnaire will focus on prices paid for fuel. The interviews would be conducted and then data collated. Whilst carrying out my research into the topic about the fuel price increase the question which came to mind was Does the fuel price increase, as a result of the recession, affect supermarkets on a day-to-day operation? In regards to this researched question, this has led to me thinking about the level of impact in which fuel prices is having on different supermarkets. Therefore this helped me choose this as my research topic fuel price increase. In terms of the research question, my research objective which is being looked to be achieved is To identify the ways in which businesses operate in terms of distribution and delivery and how this contributes to success or failure. Research into the topic of fuel prices was chosen as it is a continual business issue, relates to the economy and was an interesting topic to gain more insight into. The other research idea Should healthy eating be promoted within schools? was discarded as it does not link into business, but brings in more persuasive ideas regarding healthy eating. The research objective was to identify the ways in which healthy eating can be encouraged. This type of issue is a continuing debate and has been discussed by the government and the NHS, leading to a variety of initiatives e.g. 5 a day. Should Microsoft technology be improved for businesses? was the other research topic but was then eliminated as it already improves regularly and helps them become more up-to-date. The objective of this research idea was To investigate into the type of features preferred by businesses and any improvements they would recommend. This project takes an Inductive approach, supported by secondary data gained from published reports and scholarly work. Also, primary data was obtained through a series of telephone interviews using semi-structured questionnaires. Implementing the inductive research approach was successful for this research project as the research question acquired various perspectives. Therefore, the overall conclusion could not be drawn until the research had been analysed. One benefit of using the inductive approach was that the main categories emerge as data is collected and analysed (BPP p234-235). However there were different limitations like it being time consuming and an intensive research method which required high levels of resources to help compare different views. Having collected all the information it would therefore not reveal any results at the end based on the analysis. This type of approach involves Qualitative data as the results were not number based and was targeted at a smaller population which consisted of supermarket transport managers who would have vast knowledge of fuel prices. This therefore helps ensure my results are valid, reliable and objective. The limitation experienced was that the reliability of the data was purely dependent upon my organisational skills and analytical knowledge. As the questionnaire was my collection method alongside interviews, this meant that it helped me check the reliability of the gathered data and explore ideas deeper through the interviews. For this research topic, the survey method will be implemented and will involve preparation of a questionnaire which will be answered by the various supermarkets. Using the survey method will help obtain information which can be analysed and patterns extracted and comparisons made (Bell, 1999). Non-Probability Sampling is a sampling method which is conducted and relies on the judgement of the researcher/s in terms of selecting their respondents. The sample size for the research used is smaller and this meant it could be targeted at specific respondents. The type of sampling method carried out for the research provided was Convenience Sampling because it allowed communication to specific people within the company. This method does not require or use any sample design. This sampling method was chosen over others e.g Random, Systematic or Quota sampling method due to the reason that it helped provide useful information, as the sample was kept representative according to the organisations being investigated into. Besides this type of sampling, Convenience sampling was more relevant to my research compared to Random Sampling, which would have involved the selection of the population randomly. It was decided as the 2nd type of sampling due to the fact that it works well with small groups. The questionnaire will provide information from the perspective of supermarkets, being both consumers and suppliers. Supermarkets consume fuel as part of their distribution costs and supply fuel to customers at their petrol stations. A questionnaire was created in order to help acquire very specific information from the interviewees. Research was conducted first hand because there was no research team available. Asking specific questions will help provide precise data which can be compared and analysed in the report. However, questionnaire responses may be misunderstood and have an adverse impact on overall comparisons. Information obtained relates to the current situation and was gathered within a short span of time, resulting in brief answers. The responses from the questionnaire will be put into a bar chart to show the comparisons of how the supermarkets are operating in terms of the fuel price increase. By putting the data in the form of a bar chart, it would help summarise the large set of data easily and can be understood. However there were other methods e.g. pie charts and histograms, which were discarded as the bar charts would represent the data more easily and at the end was able to make straightforward overall comparisons. Table 1 Research Action Plan Task 17/05/12- 24/05/12 25/05/12- 31/05/12 17/09/12- 21/09/12 22/09/12- 28/09/12 01/10/12-09/10/12 08/11/12 15/11/12 16/11/12 22/11/12 Topic Selection Complete Action Plan Complete Code of Ethics Complete Data Collection Complete Questionnaire Complete Research Obj Complete Sampling method Complete Proposal completion Complete Throughout this project the milestone progress reviews took place: When the research topic was decided upon 31st May 2012 On finalisation and gathering of literature sources mentioned in the literature review When the research project proposal had been done 22nd November 2012 On completion of main report critique Code of Ethics To ensure, within this project, the content included is based on my findings To monitor and record progress on the action plan progress table Completion of the research project within the specified time limit Questionnaire responses kept confidentially and not distributed amongst other interviewees Only to use qualitative data once approved by interviewee Interviewees receive copies of the final report Advantages of surveys Disadvantages of surveys Focus on data in comparison to theory Data implication can be ignored Caters for small-scale qualitative research Questionnaires and sampling could lead to lack in depth and detail Allows use of tools like questionnaire and telephone 1:1 interviews Inaccurate or not honest responses Results obtained fairly quick Time consuming in terms of accuracy and honesty If survey structure well organised then this means the analysis is made easier Surveys provide a range Sources of data and methods of data collation It was decided that there were advantages to collecting and using further existing secondary data, such as time and cost saving. However, since the data has been acquired by someone else, it could be out of date or unreliable. Because of this, it is imperative that the research is checked to ensure it is accurate and reliable to use for the research question. The research into surveys, deduced that Denscombe (1998) had proposed pros and cons as follows: Supermarkets will have cartels Organization of the Petroleum Exporting Countries (OPEC) who will sell fuel to wholesalers e.g. Shell and then sell it to Tesco. This will mean that Tesco will be dependent upon OPEC when getting their fuel. Whilst OPEC is a cartel, this means they would be known to dominate the market and want to get suitable prices. If Tesco were to make agreements with OPEC, they would find it hard because OPEC will not receive any benefits. Overall this issue would have a big significance on Tesco because they dont have any control with aspects such as how much petrol OPEC will supply to them as well as the prices charged. Justify the chosen methodology Morrisons Faculty team was involved in the collection of the primary data. They answered the questions accurately and the information provided, led to a better understanding about their operations. This helped to obtain the correct data, which provided other essential data based around current and future actions. There are minimal academic literature based around the topic of fuel prices which can be studied in order to help draw up an overall judgement based on the findings. This is a systematic topic and has been discussed by various experts in this field. Since secondary data collation involved collecting information from that which someone else had collected, the information was reviewed before including it in this research project. It was necessary to compare the different sources of secondary information and to include it after review. http://i.dailymail.co.uk/i/pix/2012/03/16/article-0-12327C8B000005DC-223_468x343.jpghttp://www.gscape.com/images/econ/charts/WeeklyFuelPrices130212.gif UK Fuel Prices for Monday 17th September 2012 Avg Min Max Unleaded 140.05p 133.9p 152.9p Diesel 144.39p 138.9p 156.9p Super Unleaded 147.72p 139.9p 159.9p Premium Diesel 153.02p 146.9p 161.9p LPG 74.94p 67.7p 83.9p According to http://www.theupcoming.co.uk/2012/03/20/inflation-continues-fall-despite-the-rise-in-petrol-prices/ The Consumer Price (CPI) Index rate of inflation had fallen from 3.6% in January to 3.4% the following month during February. David Kern, the Chief Economist at the British Chambers of Commerce said, The marked increases in world oil and food prices since the beginning of the year are worrying and support our view that further declines in domestic inflation, both this year and next, will not be as sharp as the Banks Monetary Policy Committee. Conferring to the Daily Express on Tuesday 15th May 2012, Morrisons announced they were going to reduce their petrol prices by 2p a litre. This action being taken would lead to price wars occurring between both supermarkets and petrol stations e.g. BP, Esso and Texaco. As a result of Morrisons decreasing their fuel prices, this has led to other retailers looking to consider doing this. The main reason why Morrisons has chosen this is because it will help them stay more competitive especially in the current UK recession. With retailers like Tesco, they were considering 2p cut in prices as well as their 5p off vouchers on top. In accordance to the Telegraph published online, it mentions that The cost of oil has jumped 30pc over the last four months, raising fears that the recent easing in inflation would be temporary. Petrol prices rose 3.5p per litre during last month to  £1.35 while diesel increased 3.3p to  £1.40. http://www.telegraph.co.uk/finance/economics/9549808/UK-inflation-eases-in-August-despite-rising-fuel-costs.html It is known that having high fuel prices will mean that various industries e.g. Primary Sector, Secondary Sector, Tertiary Sector and the commercial industry who rely on any form of transportation will find themselves having to increase their prices in order to maintain their profits and be cost effective. According to the Daily Express as of Thursday 6th September 2012, it mentions about The Office of Fair trading (OFT) looking further into the fuel price increase and their inquiry will address competition issues and concerns over price co-ordination. From customers views they reckon that when the price of crude oil decreases, this is not being reflected at the pump as quick as they would like. The Daily telegraph published 21st March 2012, talks about due to both fuel prices and food, this is therefore leading to higher living costs. There have been experts who warn that the situation is unlikely to improve in the short-term. Experts have warned that drivers may either change their job or quit as a result of an increase in fuel duty. It has been proven that A hike in fuel costs at the start of the holiday season will backfire as many people are already cutting back. Jayne Atherton mentions in the Metro newspaper on Wednesday 21st March 2012 that most manufacturers are likely to raise prices because of the spiralling cost of oil. Its believed that any added rises in the cost of oil will be a significant concern. Its mentioned within the Metro newspaper dated 25th September 2012 about 2 supermarkets starting a fuel price war; Asda and Morrisons. In terms of this price war, Asda say they will cut their prices on both unleaded and diesel by 3p to 135.7p and 139.7p. Conversely Morrisons also took the same action by reducing their prices 3p. This price war will therefore lead to other supermarkets e.g. Tesco, Sainsburys, Esso, Texaco and BP, wanting to follow the same procedure. Asda had an income tracker report done in 2008 which showed the huge impact high fuel prices have on the average familys monthly budget. During August it was reported that apart from fuel prices staying low last year, the costs at the pumps had rose. Within the Daily Mail on Saturday 22nd September 2012, it says that as oil has been rising slowly over months from $100 a barrel to $115 a barrel, this has pushed petrol prices and energy bills up for different UK households. Oil prices have however fallen dramatically within previous days with fears about the slowdown in the global economy hitting demand. The metro newspaper on 10th October mentions that as cost of fuel continues to rise, this is urging motorists to stay off the roads. As motorists vanish, most are staying off the roads to help save money and also reduce their unnecessary trips. The only constant they see is the fact wages arent increasing in line with inflation but there are other costs particularly petrol and diesel that are going up. Ross McGuiness makes a decision about them having no option but to cut back and some people have cut out their cars. His study into the fuel price increase leads to the debate about how prices are increased and passed on. As argued by Brian Madderson, he mentions that when purchasing fuel which costs  £50,000, 60% of the cost is made of fuel duty and VAT. This cost is paid by small retailers to the government through the supplier before the tax is collected from the customers. This led to him believing that is absurd. Furthermore Chris Hunt, General Director at UK Petroleum Industry Association, stated that rising fuel costs are down to emerging car sales markets within China and India. Chris Hunt also argues that increasing fuel prices will decrease number of sales, but will on the other hand lead to oil suppliers across UK, not profiteering. With the 3p increase in fuel duty, it was believed that benefits would arise such as creating of over 70,000 jobs and boost economy growth by 0.2%. Sky news also states that We appreciate the Governments aspiration to reduce the deficit but know that hiking fuel duty up by 3p in January will only make things much worse. This therefore shows that the change would make the deficit greater and not help the economy become more stable. FairFuel UKs national spokesman Quentin Willson said: We have always argued that fuel duty shouldnt be the Treasurys sacred cash cow it should be used as a lever for growth. George Osborne mentioned that the current fuel price increase which was set for January is to be scrapped. This was demolished with the aim to help the pressure put on workers and families who are currently in need. This increase was also considered carefully as it would coincide with the rail fares increasing and changes to child benefits. Going ahead with the 3p rise, doesnt make sense for economic recovery and deficit reduction. Lee Boyce argues that with the petrol price decrease, it is still making consumers pay more at the pumps. The AA, goes along with Lee Boyces idea concerning the prices not dropping enough. In relation to the prices paid by customers, it was found by AA that drivers were also once again suffering from a price postcode lottery where motorists in one area were charged as much as 5p a litre more than drivers a few miles away. Before 2002, the real price of fuel had been dropping, which led to a decrease in the numbers of fuel-poor households. The real price increases since 2003, resulted in a doubling of the fuel poor from 1.2 million households in England in 2003 to 2.4 million in 2006. In the White Paper, the government predicted some possible future fuel price scenarios and their likely effect on fuel poverty numbers. The graph below illustrates this: Figure : England Households in fuel poverty Graph 1: England households in fuel poverty One of the best insights into why the UK is failing on fuel poverty comes from a statement in 1976 by an early campaigner, Marigold Johnson, when commenting on the rise in fuel prices after the first oil crisis in 1973. It was believed it was Societys failure to plan for an age of high-cost fuels. Subsequently, fuel prices have risen and at faster rate than the RPI (Retail Price Index), during every year since 2003. Increase size in 2005, 2006, 2008 has been severe. With an increase of 105% in residential fuel prices since 2000, this sets against a RPI increase of 26%, which represents a real fuel price increase of 62%. Residential fuel prices within the UK, have effectively increased by 84% between 2000 and Fe February 2009 for the average household. When the cause of a fuel price rise is international energy costs, then the problem, is, to an extent, out of the governments control. Government and OFGEM (Office of the gas and electricity markets) are trying to ensure that price increases which are passed onto the customers are justified. Governments own coffers are receiving more money from the VAT levied at 5% on rising fuel prices. It would be expected that there is recognition regarding fuel poverty policy consisting of disastrous effects in which rapid fuel prices can have on the fuel poor, particularly since 2003. The potential impact of these price increases on progress towards meeting the targets set out in The UK Fuel Poverty strategy means efforts will need to focus on finding most sustainable way of tackling fuel poverty. During 2008, world oil prices were US $145 and during 2009 went down to $39, leading to predictions about fuel price levels and fuel poverty extent being difficult. Current Fuel prices Average Minimum Maximum Unleaded 138.6p 132.9p 151.9p Diesel 143.3p 137.9p 156.9p 22.2 Domestic freight transport: by mode Great Britain percentage 1999 2000 2001 2002 2003 2004 5,6 2005 7 2006 2007 8 2008 9 2009 10 Goods moved (billion tonnes kilometres) Petroleum products Road1 ZBZP 5.0 6.4 5.8 5.2 5.5 5.7 5.5 5.6 5.1 6.5 4.8 Rail2 ZBZQ 1.5 1.4 1.2 1.2 1.2 1.2 1.2 1.5 1.6 1.5 1.4 Water3 ZBZR 48.6 52.7 43.5 51.7 46.9 46.9 47.2 37.8 36.4 36.4 36.4 of which: coastwise ZBZS 33.3 26.0 23.1 24.2 23.3 26.6 30.3 22.7 25.0 26.5 27.1 Pipeline9 ZBZT 11.6 11.4 11.5 10.9 10.5 10.7 10.8 10.8 10.2 10.2 10.2 All modes ZBZU 66.7 71.9 62.0 69.0 64.1 64.5 64.7 55.8 53.3 54.6 52.8 Coal and coke Road1 ZBZV 2.2 1.5 2.1 1.5 1.5 1.2 1.5 1.3 1.6 1.0 1.0 Rail2 ZBZW 4.8 4.8 6.2 5.7 5.8 6.7 8.3 8.8 7.7 7.9 6.2 Water3 ZBZX 0.5 0.2 0.5 0.3 0.5 0.3 0.4 0.5 0.5 0.5 0.3 All modes ZBZY 7.5 6.5 8.8 7.5 7.9 8.5 10.2 10.4 9.8 9.5 7.5 Other traffic Road1 ZBZZ 150.5 151.5 150.6 152.7 154.7 155.6 156.4 159.7 166.4 156.0 137.6 Rail2 ZCAA 11.9 11.9 12.0 11.7 11.9 12.5 12.2 11.8 11.9 11.2 11.4 Water3 ZCAB 9.6 14.6 14.8 15.2 13.5 12.3 13.3 13.5 13.9 12.7 11.9 All modes ZCAC 172.0 178.0 177.4 179.6 180.0 180.4 181.9 185.0 192.2 179.9 161.0 All traffic Road1 KCTA 157.7 159.4 158.5 159.4 161.7 162.5 163.4 166.7 173.1 163.5 143.5 Rail2 KCTB 18.2 18.1 19.4 18.5 18.9 20.4 21.7 21.9 21.2 20.6 19.1 Water3 ZCAD 58.7 67.4 58.8 67.2 60.9 59.5 60.9 51.9 50.8 49.7 48.6 Pipeline KCTE 11.6 11.4 11.5 10.9 10.5 10.7 10.8 10.8 10.2 10.2 10.2 All modes KCTF 246.2 256.3 248.2 256.0 252.0 253.0 256.8 251.3 255.3 244.0 221.3 Percentage of all traffic Road1 ZCAE 64.0 62.0 64.0 62.0 64.0 64.0 64.0 66.0 68.0 67.0 65.0 Rail2 ZCAF 7.0 7.0 8.0 7.0 7.0

Hong Kongindia Command and Market Economies :: essays papers

Hong Kongindia Command and Market Economies The world is run with economic systems. Two of these economic systems are the market economy and the command economy. These economic systems both greatly differ and are similar in there ways of controling the factors of production, ownership, and their incentives for efficient production. An area of the world that has a command economy is India. The government controls and makes decisions with the factors of production. The factors of production are the resources used to produce goods and services, such as lonad, labor and capital. Within these socialist areas, there is no presence of motivation, a very important factor in an economy. This is because no matter what they do for their economy, everyboyd will recieve the same pay. For example, a doctor would get paid the same amount as a dishwasher.This creates unmotivation because there is nowhere to progress to. People don’t want to work hard for the same pay as someone who doesn’t. Also, since the government controls the factors of production, and not the individual consumers themselves, it is impossible for the few in power to know the many needs, and conditions of resource availability, within the whole economy. Therefore, many things of importance are not recognized, or over looked and the economy and people suffer from the loss. It is present in these areas of socialism that many of the people suffer from poverty because of this economic system. Hong Kong used to be a command economy. Hong Kong was very poor. This poor area quickly changed over the years into a very prosporous place. This is due to it’s change to the market economy. Hong Kong is also the freest of economies. The United states of america also has a market economy. In a market economy, or free enterprise economy, the individuals own the factors of productin, and make the decisions with them. This is unlike the command econly becaues their government controls the factors of production. The market economy is more effiecent because the individuals decide for themselves how to answer economic questions, looking out for their own personal best interest. This creates motivation, in which the command economy lacks. Entreprenurs also play a large roll in the economy. In Hong Kong, it is extremely easy to start your own business.

Ethical Business in Global Trade Essay

Ethical Business in Global Trade Introduction            Ethics can be defined as the â€Å"well founded standards of right and wrong that prescribe what humans ought to do, usually in terms of rights, obligations, benefits to the society, fairness or specific virtues† (Manuel et al., 2010) In relation to Human labor, Auret van Heerden (2010) says that employers should treat employees like Human beings. Violation of human rights in the global food chain amounts to unethical business practices. Employees working for excess hours, companies employing children below the International Labor Organization required age, employees being paid peanuts and safety measures for employees not being maintained to avoid injuries are some of the unethical practices related to global chain            The role of the governments in the supply chain is to ensure that the rights of workers who work hard in production of raw materials and preparation of the final products are maintained (Auret, V.H. 2010). These governments are also placed with a duty to ensure that the safety of the products is maintained. This is possible with establishment of a code of conduct to govern operations of the companies involved in the global food chains. Frequent Inspection should be conducted with stiff penalties to companies that violate the human rights and safety measures of the product.            Management of a global market presents a challenge due to involvement of many governments. While the USA may have well laid down measures and regulations for ethical business practices of these companies, other countries in the global food chain may not have them. Some countries avoid putting up stringent measures in order for them to attract investors at the expense of ethical business activities            The fight for ethical business practices in the global food chain is very much achievable. These will require governments to work together. Consumers of these products can also have a critical role to play by giving a snub to products from companies that do not conform to ethical business practices. References Auret van Heerden (2010). Making Global Labor Fair. Retrieved fromhttp://video.ted.com/talk/podcast/2010G/None/AuretvanHeerden_2010G.mp4Manuel, V., Claire, A., Thomas, S., S. J., & Michael, J.M. What is Ethics? Retrieved from http://www.scu.edu/ethics/practicing/decision/whatisethics2.html?utm_expid=6222598-1.kwhpOqkUTTeXUGI-lneo5w.2 Source document

Legalization Of Recreational Marijuana In The California Assignment

Legalization Of Recreational Marijuana In The California Assignment Legalization Of Recreational Marijuana In The California – Assignment Example To the editor of San Francisco Chronicles: Over the past few months, there has been a wide proposal on the legalization of Marijuana in the of California. This is a ground-breaking piece of law that each American citizen should back right now. Many citizens do not realize it, but illegalizing the sale of marijuana is one of the main reasons why many people are using it. In fact, a majority of individuals using marijuana purchase more of this substance in the ‘black market’. If this proposed legislation is put into effect, it will not only ensure that the citizens do not abuse the highly important weed but it will enable the government to increase its revenue from taxes as well. Legalization of pot will be a major step in the development of California. Findings demonstrate that marijuana has multiple advantages. The illegalization of marijuana is also a gateway to other hard drugs such as heroin and cocaine that are more dangerous to the human body. Besides, legalizing Marijuana will cause a consecutive decline in the abuse of drugs such as alcohol and tobacco which cause even more deaths in America than pot. If marijuana is legalized, education will be provided on its long term side effects and therefore many people will not abuse it. The legalization of Marijuana bill has been proposed, and it now needs to be approved. Your voice will be of essential important in this matter. We would like to embolden you to write or talk to the Congressman and let him know that we support this proposed bill. The more support the bill gets, the more likelihood that it will be approved into law. Sincerely, Name Position/ Occupation Address

Understanding the Ninth Amendments Effect and History

Understanding the Ninth Amendments Effect and History The Ninth Amendment to the U.S. Constitution attempts to ensure that certain rights - while not specifically listed as being granted to the American people in the other sections of the Bill of Rights - should not be violated. The complete text of the Ninth Amendment states: â€Å"The enumeration in the Constitution of certain rights shall not be construed to deny or disparage others retained by the people.† Over the years, the federal courts have interpreted the Ninth Amendment as confirming the existence of such implied or â€Å"unenumerated† rights outside those expressly protected by the Bill of Rights. Today, the Amendment is often cited in legal attempts to prevent the federal government from expanding the powers of Congress specifically granted to it under Article I, Section 8 of the Constitution. The Ninth Amendment, included as part of the original 12 provisions of the Bill of Rights, was submitted to the states on September 5, 1789, and was ratified on December 15, 1791. Why This Amendment Exists When the then proposed U.S. Constitution was submitted to the states in 1787, it was still strongly opposed by the Anti-Federalist Party, Led by Patrick Henry. One of their main objection to the Constitution as submitted was its omission of a list of rights specifically granted to the people - a â€Å"bill of rights.† However, the Federalist Party, led by James Madison and Thomas Jefferson, contended that it would be impossible for such a bill of rights to list all conceivable rights, and that a partial list would be dangerous because some might claim that because a given right was not specifically listed as protected, the government had the power to limit or even deny it. In an attempt to resolve the debate, the Virginia Ratifying Convention proposed a compromise in the form of a constitutional amendment stating that any future amendments limiting the powers of Congress should not be taken as justification for expanding those powers. This proposal led to the creation of the Ninth Amendment. Practical Effect Of all the amendments in the Bill of Rights, none is stranger or harder to interpret than the Ninth. At the time it was proposed, there was no mechanism by which the Bill of Rights could be enforced. The Supreme Court had not yet established the power to strike down unconstitutional legislation, and it was not widely expected to. The Bill of Rights was, in other words, unenforceable. So what would an enforceable Ninth Amendment look like? Strict Constructionism and the Ninth Amendment There are multiple schools of thought on this issue. Supreme Court justices who belong to the strict constructionist school of interpretation essentially say that the Ninth Amendment is too vague to have any binding authority. They push it aside as a historical curiosity, in much the same way that more modernist justices sometimes push the Second Amendment aside. Implicit Rights At the  Supreme Court level,  most justices do believe that the Ninth Amendment has binding authority, and they use it to protect implicit rights hinted at but not explicated elsewhere in the Constitution. Implicit rights include both the right to privacy outlined in  the landmark 1965 Supreme Court case of  Griswold v. Connecticut, but also basic unspecified rights such as the right to travel and the right to the presumption of innocence until proven guilty.   Writing in the Court’s majority opinion Justice William O. Douglas stated that â€Å"specific guarantees in the Bill of Rights have penumbras, formed by emanations from those guarantees that help give them life and substance.† In a lengthy concurrence, Justice Arthur Goldberg added, â€Å"The language and history of the Ninth Amendment reveal that the Framers of the Constitution believed that there are additional fundamental rights, protected from governmental infringement, which exist alongside those fundamental rights specifically mentioned in the first eight constitutional amendments.† Updated by Robert Longley

Day of Infamy by Walter Lord essays

Day of Infamy by Walter Lord essays (New York: Henry Holt Walter Lords Day of Infamy traces the drama of the massive aerial attack of Pearl Harbor on the morning of December 7, 1941. In this book, Lord painstakingly reconstructed not just the why and the way of the attack, but also how it happened, how people could have been so unaware of what might happen, and the slowness to regroup when it did. He begins with the innocence (and evils) of the night before the tragedy. He ends with the famous national radio address of President Franklin D. Roosevelts speech before Congress the following day. Lord is also known for his bestselling book A Night to Remember, which was written in the same style as this book, with minute-to-minute accounts of the sinking of the Titanic. Walter Lord reminds the reader of just how innocent people are in the moments before history is changed forever. How untrained they are at putting the clues together, and just how unprepared they can be, until after the fact- when they become fiercely patriotic and regroup. He does not spend a lot of time pointing fingers or placing blame, but remains on the raw human experiences of the day. As author James Michner wrote in The New York Times, It stuns the reader with the weight of reality. Lord shows the way Americans believed that no one had the ability to reach them, let alone attack the U.S. This is obvious in Chapter VIIs title: I Didnt Even Know They Were Sore At Us!, p 64. He illustrates how everyone was oblivious to extreme nature of events going on around him or her, even after the bombs were dropped. Lord goes into meticulous detail to recount the day all the way from the Japanese build up of a secret mission, to the Americans living and stationed on the Hawaiian Island of Oahu. He interviewed 577 people to recount the events from as m ...

Maria Gaetana Agnesi Research Paper Example | Topics and Well Written Essays - 500 words

Maria Gaetana Agnesi - Research Paper Example The mathematic books of Agnesi contain deeper explanations and numerous examples. The books contain various methods of tackling a problem and numerous examples of the same kind question. However, the language in use is simple for learners to understand the mathematical concepts easily and encourage love of mathematics in general. Most of the textbooks currently only have few examples and a formula to solve a problem in a particular topic. This serves as a great disadvantage to students as mathematics as a subject requires numerous examples as well as different ways of solving a question (Cupillari) Agnesi had the knowledge of different languages and therefore, coming up with a simpler mathematical language was easy for her. She simplifies her books into easy language containing important details explaining the derivation of a particular mathematical formula and most importantly how to use the formula to solve a mathematical problem in the specific topic providing better option to other textbooks. Textbooks of Agnesi contain many examples in each topic providing adequate self-assessment questions for the students. The best textbooks should contain numerous examples on how to make use of a mathematical formula to solve a question. Students using Agnes i’s textbooks acquire higher grades due to the many examples in provision comparing to students using other textbooks. Numerous alternative methods are in provision in Agnesi mathematics textbooks in comparison to other mathematics textbooks. It provides choices to students on which method to master depending on the understanding of the question and the knowledge of the concept to use. This provides the opportunity of choosing a particular way of doing a question, which is rare in other textbooks (Baltimore) A cubic curve refers to an algebraic curve with the specific curve order 3. The form of shows a cubic function a cubic function is clearly shown by a polynomial function of degree 3. If

MrsFields case study Example | Topics and Well Written Essays - 1250 words

MrsFields - Case Study Example LPB was a large company; their formulations of baking from the frozen dough the croissants, bread, hot soups and sandwiches, taken from Fields extended cookies business, had a different style. Technological processes at Fields were not changed to make adjustment with the working of LPB. Fields wanted to get the benefit of their in-practice MIS system, which was not customized to assimilate the management functions of LPB like Fields had done when it acquired Chocolate Chip Company, and made changes in the MIS system. Fields assumed that that it would be able to benefit from its current MIS system for adapting different overhead functions of PBS. Working in a totally different management hierarchy would not have been easy particularly when no job orientation was given. Things could have gone haphazard without any staff reporting to the senior level, affecting performance in all segments of the organization. As per Randy’s statement, the organizational structure at LPB was not similar to the functioning at Fields who has been using IT to manage business functions across the globe efficiently. The impact of discharging 50 human resources from their duties of the acquired company and without providing the training of the work processes of the new organization structure, I would have felt crippled as a manager. Reporting of store managers at Fields was straight to all senior and middle level managers culminating in final reporting to Debbie and Randy. The change for the store manager’s rank was sudden from a traditional way of functioning to remaining available for seamless exchange of information horizontally and vertically on the vast communication network. Chances of failure had been more than success as store manager. In my opinion, it would be wrong to say that business at LPB was same in kind as it was at Fields in different aspects like products, customers, and business challenges. Although bakery was the major business at LPB but it offered other

Ayurvedic Medicine Used as Alternative Therapy in Modern Medicine Essay

Ayurvedic Medicine Used as Alternative Therapy in Modern Medicine - Essay Example This was traditional medicine to treat fever, cough and diarrhea. People were encouraged to suppress natural urges which was considered as unhealthy by moderating the amount and quantity of food intake, sleep and sexual intercourses to ensure healthy body at all times. Gregory (2004) states that alternative medicine refers to treatment of diseases using methods such as massage therapy or herbal use. This practice is being highly appreciated by learning institutions who teach about it. Importance Frawley and Ranade (2001) states that herbal method as one of the alternative method has been well embraced by many communities in the world hence rendering the use of Ayurvedic medicine easy and of great importance. People consider this method efficient since it is less costly and also has few side effects. This method also have seem to treat a variety of diseases which lead people to prefer using it since he will only require one drug to heal so many diseases. This alternative is readily av ailable since most of these herbs are found free of charge in the forests and other bushy areas within the society. This method has been considered to be less demanding because it does not require highly qualified personnel to administer the medicine. This method is also effective since it is not selective with the age bracket hence can be used by all people within the community. Unlike the artificial drugs these medicines do not lead to very adverse side effects. It does not have the time limits hence one can use it as long as he wants. This therapeutic method is easily acceptable to be used by people because it is mostly orally administered which is the most preferred method. This method can not react while using other methods hence gives it another advantage of it to be considered above the other method. This method is reliable since one can get it from various places. Frawley and Ranade (2001) argues that this method is used even by the illiterate people in the society since the instructions given are clear and simple to be understood hence very low cases of overdosing. Since this method is given by people who are locals in the community the patients respect the instructions given no deviations since there is sense of belonging. At times there is reinforcement of this method by the religious people and other important people in the society hence the patients look forward to embrace this method to the fullest. Lad (2012) says that Ayurvedic medicine does not demand one to go to hospital for the treatment he can get the treatment while at home. Diagnosis According to Lad (2012) the patient who was recommended for use of this drug was to be carefully examined and undergo through various diagnosis procedures using all body senses by the health profession administering the drug; since each component of the body had the ability to influence others either negatively or positively. This aspect was to be carefully considered during the drug diagnosis and therapy. V arious things were to be checked such as the body composition, any disorder, body stability and all other body systemic functions. The systemic composition and functioning was checked so that to prevent any adverse side effect that would arise while on medication. The body was to be considered compatible to the drug for one to be in a position to use the drug. There were various criteria that were

Domestic Terror in the United States Term Paper

Domestic Terror in the United States - Term Paper Example ive of excessive force, there must be pressure applied to others in society or government or otherwise persuasive agendas, and the objectives of these acts must be linked with sociological or politically-motivated agendas (FEMA, 1997). Domestic terrorism is characterized as a holistic blend of the aforementioned objectives and activities, which occur within a national region, which are conducted against social systems and ideologies, governmental actors, and citizenry communities. Some of the most common terrorist activities occurring domestically include implementing a variety of illegal pursuits so as to fund hostile or brutal objectives, as well as using a variety of methodologies to elicit threats against government and society so as to establish support or compliance with a specific political or social agenda. The composition of domestic terrorism is complex and usually consists of multi-faceted objectives of the terrorist party or group which has substantial implications for local, state and federal-level institutions when attempting to contend with terrorist activities and agendas. It often involves a series of complicated analyses and interventions within many diverse federal institutions under a collaborative model with local and state-level authorities to achieve terrorist reduction and opposition. This essay examines a variety of domestic terrorism attacks that have occurred in the United States, focusing on the politically-motivated or socially-motivated objectives that drove these violent acts. The project further examines the threats and dangers of domestic terrorism today as opposed to international terrorism within U.S. borders and provides an analysis of the programs currently in place at the federal, state and local levels to prevent reoccurrences of domestic terrorism. It is significant and prudent for American citizens and empowered institutions to understand the dynamics of what serves as catalysts for domestic terrorism as domestic

Multinational buainess Essay Example | Topics and Well Written Essays - 2500 words

Multinational buainess - Essay Example At the present economic environment such theories and models are considered to be very important as the importance of multinational organizations to the global economy has increased significantly. Dunning’s eclectic paradigm theory is one of the most widely discussed theories in the world of international production. This paper attempts to present a clear description of this theory and several of its dimensions. The paper also includes a proper analysis of the criticisms that the Dunning’s model has faced over the years. There are several other theories on the similar subject and Dunning’s theory can be compared with those to analysis the effectiveness of it. This paper includes such a comparative analysis that is likely to help an individual to have a clear conception regarding the entire picture of such theories. Academic Literature John H Dunning is famous in the international business arena for his ‘Eclectic Paradigm’ theory. It is actually the i ntegration of several theories on internationalization. It covers many mainstream theories such as Ohlin and Heckscher’s ‘factor endowment theory’, Coase’s ‘trasaction cost theory’, Hymer’s ‘monopolistic advantage theory’ and ‘internalization theory’ of Casson and Buckley. In 1976, Dunning presented the core concept of eclectic paradigm at Nobel Symposium which is located in Stockholm. The theory explains precisely why a firm should go global. Dunning attempted to provide a clear and complete explanation regarding international production. He has consistently supplemented and improved his theory in accordance to various developments in the global economic environment and also in accordance to several constructive criticisms. Dunning opined that there are three conditions that have significant influence over the firm’s decision regarding transnational production. In other words, a firm will involve in â⠂¬Ëœinternational production’ if these three conditions are met. The three conditions are 1) ‘ownership-specific advantages’, 2) ‘internalization advantages’ and 3) ‘location specific advantages’ (Dicken, 2007). Each of them is properly described below. Ownership-Specific Advantages According to Dunning, an organization must have some ownership specific advantages which are not possessed by the rival organizations of other countries. The basis of this theory of Dunning lies in the ‘Monopolistic Advantage Theory’ that was developed by Hymer. According to Dunning, ownership specific advantages â€Å"must be sufficient to compensate for the costs of setting up and operating a foreign value-adding operation in addition to those faced by indigenous producers or potential producers† (Dunning, 1981). These advantages are like ‘internal assets’ of a firm which is the sole owner of them. Ownership specific advanta ges can be gained from the factors like technology, knowledge, internal brands, human resources etc. (Dicken, 2007). In the words of Dunning, ownership specific advantages â€Å"need not arise from multinationality† (Perry, 1992). These advantages

Asian American and Asian communities in U.S Essay

Asian American and Asian communities in U.S - Essay Example One of the very important reasons is that of something called as â€Å"Asian parents Syndrome† (Online Education Articles, n.p). It has been an observation that Asian parents generally have higher expectations of their children. It is because of these expectations that many college students feel the need to achieve greater success. Asian parents cannot simply compromise that their children are not the best among everyone, and therefore, this drives the children to work harder (Online Education Articles, n.p). Asian culture and its expectations therefore enable the Asian students to work harder. Another factor, which plays a very important role, is that of maternal expectations of children. The study conducted by Stevenson group was based on Japanese and American students and mothers. When children did not perform well in school, almost 42 percent of American mothers were satisfied with their child’s performance. Contrary to that, only five percent of Japanese mothers ra ted the performance to be satisfactory. American mothers were also willing to accept a child’s low performance. Therefore, the study concluded that such American attitudes provided an excuse for the children for not working very hard. Such attitudes continue throughout life, therefore, American students might not perform better in colleges. Asian students are very adaptable to levels of hard work (Wray, pp. 57). Historically, Asian students have been driven to work harder. Educational achievements had become very important, especially in the case of Japanese and Chinese students. A study done by Suzuki in 1977 (Nakanishi & Nishida, pp.140) showed that Asian Americans came to pursue education because they were considered a ‘minority’ group. This led them to strive harder. During the 1940s, Asians were discriminated by the trade unions. They were refused membership. After the Second World War, there was a need for white-collar employees because of technological adv ancements. Thus, education became the prime goal for Asian individuals in order to achieve success. In addition, Connor in 1975 gave the explanation that Asian students had been denied in social and extra curricular activities before the First World War. This had emphasized educational values. Therefore, studies have proved that historically, Asian students have had the motive to pursue education with full zeal (Nakanishi & Nishida, pp.140). A study conducted on Japanese students and teachers showed that teachers expected a greater level from their students. They are taught that education is difficult, not always exciting, and it also requires a great amount of hard work. Responsibility and perseverance is also taught. Hence, Japanese students have been socialized to achieve success through education. Therefore, this attitude remains with them throughout life, and therefore when they go to study in colleges in US, they will tend to perform better. Therefore, values of culture also p lay a significant role (Wray, pp. 54). Generally, Asian immigrants also have spent more amount of time in school. On average, Asian students in their countries spent an average of 240 days in school per year, while the American students spent around 178 days in school every year. Therefore, this increases the performance of the Asian students (Santrock, pp. 626). Some other cultural reasons, which might also form part of religious values, attempt to explain why Asians generally do better in colleges. There are certain cultural values, which are common

Domestic Terror in the United States Term Paper

Domestic Terror in the United States - Term Paper Example ive of excessive force, there must be pressure applied to others in society or government or otherwise persuasive agendas, and the objectives of these acts must be linked with sociological or politically-motivated agendas (FEMA, 1997). Domestic terrorism is characterized as a holistic blend of the aforementioned objectives and activities, which occur within a national region, which are conducted against social systems and ideologies, governmental actors, and citizenry communities. Some of the most common terrorist activities occurring domestically include implementing a variety of illegal pursuits so as to fund hostile or brutal objectives, as well as using a variety of methodologies to elicit threats against government and society so as to establish support or compliance with a specific political or social agenda. The composition of domestic terrorism is complex and usually consists of multi-faceted objectives of the terrorist party or group which has substantial implications for local, state and federal-level institutions when attempting to contend with terrorist activities and agendas. It often involves a series of complicated analyses and interventions within many diverse federal institutions under a collaborative model with local and state-level authorities to achieve terrorist reduction and opposition. This essay examines a variety of domestic terrorism attacks that have occurred in the United States, focusing on the politically-motivated or socially-motivated objectives that drove these violent acts. The project further examines the threats and dangers of domestic terrorism today as opposed to international terrorism within U.S. borders and provides an analysis of the programs currently in place at the federal, state and local levels to prevent reoccurrences of domestic terrorism. It is significant and prudent for American citizens and empowered institutions to understand the dynamics of what serves as catalysts for domestic terrorism as domestic

Asian American and Asian communities in U.S Essay

Asian American and Asian communities in U.S - Essay Example One of the very important reasons is that of something called as â€Å"Asian parents Syndrome† (Online Education Articles, n.p). It has been an observation that Asian parents generally have higher expectations of their children. It is because of these expectations that many college students feel the need to achieve greater success. Asian parents cannot simply compromise that their children are not the best among everyone, and therefore, this drives the children to work harder (Online Education Articles, n.p). Asian culture and its expectations therefore enable the Asian students to work harder. Another factor, which plays a very important role, is that of maternal expectations of children. The study conducted by Stevenson group was based on Japanese and American students and mothers. When children did not perform well in school, almost 42 percent of American mothers were satisfied with their child’s performance. Contrary to that, only five percent of Japanese mothers ra ted the performance to be satisfactory. American mothers were also willing to accept a child’s low performance. Therefore, the study concluded that such American attitudes provided an excuse for the children for not working very hard. Such attitudes continue throughout life, therefore, American students might not perform better in colleges. Asian students are very adaptable to levels of hard work (Wray, pp. 57). Historically, Asian students have been driven to work harder. Educational achievements had become very important, especially in the case of Japanese and Chinese students. A study done by Suzuki in 1977 (Nakanishi & Nishida, pp.140) showed that Asian Americans came to pursue education because they were considered a ‘minority’ group. This led them to strive harder. During the 1940s, Asians were discriminated by the trade unions. They were refused membership. After the Second World War, there was a need for white-collar employees because of technological adv ancements. Thus, education became the prime goal for Asian individuals in order to achieve success. In addition, Connor in 1975 gave the explanation that Asian students had been denied in social and extra curricular activities before the First World War. This had emphasized educational values. Therefore, studies have proved that historically, Asian students have had the motive to pursue education with full zeal (Nakanishi & Nishida, pp.140). A study conducted on Japanese students and teachers showed that teachers expected a greater level from their students. They are taught that education is difficult, not always exciting, and it also requires a great amount of hard work. Responsibility and perseverance is also taught. Hence, Japanese students have been socialized to achieve success through education. Therefore, this attitude remains with them throughout life, and therefore when they go to study in colleges in US, they will tend to perform better. Therefore, values of culture also p lay a significant role (Wray, pp. 54). Generally, Asian immigrants also have spent more amount of time in school. On average, Asian students in their countries spent an average of 240 days in school per year, while the American students spent around 178 days in school every year. Therefore, this increases the performance of the Asian students (Santrock, pp. 626). Some other cultural reasons, which might also form part of religious values, attempt to explain why Asians generally do better in colleges. There are certain cultural values, which are common

Toxic Epidermal Necrolysis Essay Example for Free

Toxic Epidermal Necrolysis Essay Toxic Epidermal Necrolysis can be a life threatening disease, which was first discovered in 1922 by pediatrician A.M. Stevens and S.C. Johnson. Toxic Epidermal Necrolysis, also referred to as TEN disease, is usually a result of a drug reaction. TEN disease is not just limited to any one part of the world or group of people, but reaches all corners of the world. TEN symptoms can start out as a simple cough, headache or a fever. At this point if it is TEN disease the following, symptoms can be a rash turning into blisters across the face and parts of the body. Treatments for those who survive are given in the hospital. At this point, your Doctor will have taken you off any medications that may be causing this infection. The frequency of Toxic Epidermal Necrolysis in the United States is estimated to be between 0.22-1.23 cases per 100,000 people per year. However, in HIV-positive people the rate of the disease increases to one case per thousand per year. Additionally, the rate of infection in Germany and France is reported to be between 1 to 1.9 cases per million people. The worldwide frequency of TEN is 0.4 to 1.3 cases per million annually. Regardless of world location, TEN can affect any age group, but it appears to be more frequent in people age 46 to 63 years old. TEN is more prevalent in the elderly population because they tend to use more medications, thus increasing the rate of infection. On the other hand, unlike the elderly, children tend to contract TEN by infection rather than medication reactions. Furthermore, the blood relatives of those who have been affected by TEN can be at a risk of developing the disorder because heredity is suspected in skin disorders caused by drug reactions. Therefore, blood relatives of the infected person must not use the medication that is thought to be the cause, and they should be cautious when using similar medications. The first step of treatment is to stop taking the medications that can bring on TEN. While this may help prevent further complications, treatment will be painful and take some time. One who is treating a patient with Toxic Epidermal Necrolysis main job is to keep infection down and to set up IVs for fluids and nutrition so that the body has the capability to start the healing process. As stated earlier, treatment may take some time so to keep the patient as comfortable as possible, pain medication such as morphine may be administered. Treatment for Toxic Epidermal Necrolysis is usually done in the ICU, and is referred to more as supportive care, which means the ICU unit tries to create a positive atmosphere for the body to fight and heal its self of this attack. In conclusion, Toxic Epidermal Necrolysis may not be one of the most common diseases out there, but it would behoove one to know the symptoms, such as a rash or blisters appearing or a persistent fever and possible blisters forming in the mouth and eyes. Many confuse this beginning outbreak as the flu. Therefore, if someone you know starts having a reaction that resembles TEN, then one should seek medical help before the body is plagued with an all-out reaction that resembles the blisters of a burn victim.

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