Essay Hoosiers a Film Review - 1283 Words

HOOSIERS A film review Hoosiers is a film about second chances. Redemption is given to a short tempered coach, who was issued a lifetime suspension by the NCAA for physically assaulting a player, and a former star player- turned town drunk. These two defeat their odds by taking a small town high school basketball team from being just 15 and 10, all the way to the state championship. In my review, I’ll attempt to explain how this coach matches up against Kouzes and Posner’s â€Å"Five Practices of Exemplary Leadership.† Our setting is rural Indiana in a town called Hickory. It’s a place that’s resistant to change. Hickory is a place where, according to Myra Fleener, a character in the film, â€Å"basketball heroes are treated like gods†. This†¦show more content†¦They must clarify values. Norman Dale valued teamwork. He believed that no player was more important than the other and he expressed this during practice and showed it by changing the way the Hicko ry Huskers practiced. Unlike the town who wanted to get their star player back and keep the basketball in his hands and not confuse the boys by changing their routine, the new coach wanted to break these barriers. He â€Å"modeled the way† by sticking to his values and ignoring the pestering from the fathers and other townsfolk who invited themselves to practice and by kick out players who did not want to listen during practice. Leaders gaze across the horizon of time, imagining the attractive opportunities that are in store when they and their constituents arrive at a destination. They envision exciting and ennobling possibilities. The Leadership Challenge, pg. 17. Inspiring a Shared Vision comes second on Kouzes and Posner’s list. Coach Dale had obtained previous success and knew he could make the Hucksters better. His vision was to go back to the fundamentals of basketball and playing better defense was part of it. Norman wasn’t about scrimmage or a whole lot of shooting during practice. His vision involved passing the ball no less than four times before taking a shot. This enabled his strong value of teamwork to take place. He tried to inspire others toShow MoreRelatedRemember The Titans By Gregory Allen Howard Essay1207 Words   |  5 PagesRemember the Titans is a 2000 non-fiction set in the early 1970’s. This well-known film would feel familiar to anyone that has ever seen a racially divided group or team dealing with the dislike from the public. For example, the film Hoosiers and Miracle on Ice. Remember the Titans was about a renowned football team that was dealing with segregation. T.C. William High School’s board was forced to have blacks infiltrate an all-white school. The foundation of footballs great tradition was put to the

Lifestyle Sustainability and the Environment Essay examples

Lifestyle Sustainability and the Environment In a perfect ecosystem everything gives and takes equally, and the cycle of life is sustained perpetually. Our current lifestyle is not environmentally sustainable. We consume more and more of the earths resources and give very little, if any, in return. The Brundtland Commission defines lifestyle sustainability as being development that seeks to meet the needs and aspirations of the present without compromising the ability to meet those of the future. Many factors are contributing to how humankind uses the earths resources and how humankind views the goal of sustainability. Public opinion, government intervention, and manufacturers seem to have the largest influence in determining how†¦show more content†¦Around the world we see groundwater supplies being depleted, agricultural soils being degraded, oceans being over-fished, oil reserves being drawn down, and forests being cut faster than they can re-grow, (Nebel and Wright, 2000, p. 6). The largest percentage of the worlds population lives in developing countries. These developing countries oftentimes have not established environmental controls in their manufacturing and farming techniques. For example, the use of DDT, long since banned for use in the United States, still sees widespread use in many developing Nations. The use of DDT is dangerous not only to insects, but also to the entire food chain, of which man is unavoidably a part. This is not to say that developed countries are not polluting. In fact, many times it is the developed countries that are exploiting the environment intentionally for personal or commercial gain. The U.S., for example, is the worlds number one producer of garbage and industrial waste. The 6% of the worlds population living in the U.S. uses an estimated 25 to 50% of the worlds nonrenewable resources and produces about 15 to 40% of the worlds waste. (Sustainable America, 1996:143) Despite the fact that most countries do have constraints and limits set for pollution, the y are still a long way from being environmentally friendly. Pollution comes in many forms: air, water, and land. Air and water resources can and should be considered global resources. The misuse of theseShow MoreRelatedSustainability And The Current Global Development Model1680 Words   |  7 PagesNations Panel on Global sustainability in January 2012, which encourages sustainable actions. Moreover, this report advises people to use their human rights to pursue sustainability and encourages governments to take the proper initiatives to resolve current day issues. Currently, social issues such as, poverty, deforestation, global hunger, and climate changes are all leading factors of our failing environment. This report along with other sources portray sustainability as a social solution involvingRead MoreHow Many Planets Worth Of Natural Capital Do We Require For Everyone You Live Like You?1101 Words   |  5 PagesQuestion 1 a) How many planets worth of natural capital do we require for everyone to live like you? According to Appendix A, if every individual had my lifestyle, two planets would have been required for the adequate provision of resources. Surprisingly, to support my needs, an emission of 7.4 tons of carbon dioxide as well as 4.2 global hectares of the Earth’s productive land would be needed. The entailed land compromises of: †¢ 48% of Energy land (i.e. land required to accommodate my carbon emission)Read MoreWhat Can Raise Awareness Of Sustainable Consumption Of Textiles For College Students1637 Words   |  7 PagesIntroduction: Many consumers aren’t educated on the importance of sustainability of clothing and textiles. The media mistakenly represents sustainability and the influence it can have on the environment. This research paper will attempt to address the question of what could raise awareness of sustainable consumption of textiles for college students. Consumers should all be aware of how they can conserve the environment by preserving clothing which is a concept that many people have not acknowledgedRead MoreAn Article About A Sustainable Living962 Words   |  4 PagesSustainable Living There is considerable discussion about sustainable living. Open up any newspaper or magazine on any given day, and you will likely see an article about the drought, food system, or global warming. It’s a hot topic. But what does sustainability mean to black folks. Some may say that we have limited involvement in the discussion. If that’s true, is it because we are focused on job security, considering that black unemployment is almost double that of whites? Are we focused more on physicalRead MoreSustainability Is Becoming An Issue Of Critical Importance For Designers And Society1549 Words   |  7 PagesSustainability is becoming an issue of critical importance for designers and society as a whole. It is important to pay attention to the factors that impact negatively on our environment by balancing economic, environmental and social dimensions as they interact and converge in the current crisis of our unsustainable civilisation (Wahl and Baxter 2008). It is critical for designers of today to be more aware and understand the importance of the material choices and resources used, as well as puttingRead MoreSustainability Position Paper : Natural Resources And Technology1652 Words   |  7 PagesSustainability Position Paper Natural resources and Technology Sarah Manzoor AB975 â€Æ' Introduction Sustainability has normally been defined as how biological systems remain diverse and productive (What is sustainability and why is it important?, 2016). However, in today’s world it refers to the need to develop methods for the earth to survive. We live in a developed world where natural resources as being consumed very fast and this is something that needs to change. Sustainability is also about thinkingRead MoreThe Sustainability Strategy of Virgin Groups Corporate1497 Words   |  6 Pagesin demographics and society in general have combined to make sustainability a mainstream issue. â€Å"The goal of sustainable development is to meet the needs of the present without compromising the ability of future generations to meet their own needs.† (G3.1 Guidelines, 2011, p4) This study report on a survey of the Virgin Groups sustainability report that summarize the Virgin companies’ sustainability strategy, evaluating the sustainability report based on the Global Reporting Initiative (GRI) principleRead MoreSustainability Multiple Choice Questions1540 Words   |  7 PagesChapter 23 Multiple Choice Questions Name: ____________________________________________________________ 1) The term sustainability refers to ________. A) maintaining resource use at current or higher levels B) keeping the natural environment and human society in a happy, healthy and functional state C) holding or increasing the current quality of human life D) always focusing on fulfilling short‑term needs E) opposing change from currentRead MoreA Study On The Sustainability Cluster Essay1271 Words   |  6 Pagespoint in life, one must decide on what one would like to do in their future. My studies would focus on detail studies in the sustainability cluster. Nowadays, sustainability is a topic which research is still maturing and design relative new. My research would emphasize in ruralization of current urban cities that would help increase innovative design presented in urban environment in Japan. Many major cities in the US and around the world have developed such that much of the land has been converted intoRead MoreImpact Of Sustainability On The Environment1715 Words   |  7 Pages‘Sustainability’, a word which everyone is coming across more often than ever before in today’s world. Yet, when it comes to defining and understanding this word, we observe various definitions and interpretations across people of various backgrounds. The question ‘What is sus tainability?’ can be compared with the question ‘What is life?’. For some, life is a physical entity between birth and death, for others it the a non physical entity of way of living and so on. Similarly, Engineers, Architects

Building a Local Fashion Brand Free Essays

Executive Summary This report is prepared on the basis of a survey on a local fashion brand (Dishes Doss), it’s a survey to building this brand as a strong brand. The concept of branding has been around for centuries as a meaner to distinguish the goods of one producer from those of another. In fact the word brand is derived from the Old Norse word brander, which meaner â€Å"to burn,† as brands were and still are the meaner. We will write a custom essay sample on Building a Local Fashion Brand or any similar topic only for you Order Now Brands identify the source or maker of a product and allow consumers to assign responsibility to a particular manufacturer or distributor. Most important, brands take on special meaning to consumers. Because of past experiences with the product and its marketing program over these years, consumers find out which brands satisfy their needs and which one do not. As a result, brands provide a shortened device or meaner of simplification for their product decisions. Building a brand is a step by step process. There are four steps or six blocks to build up a brand. Here we applied all the steps or blocks to find out the present brand situation of Dishes Doss. In order of these steps or blocks we made a questioner of 69 questions ; held a survey on 50 people. After the survey we convert the result onto percentage and put it into graph to show the situation. The topic has been selected to building some local fashion brands (dishes Doss). A major objective of the report is to make customer based equity for these brands. After studying the steps of a brand building, we surveyed the information of these brands, on the basis of brand building. 1. 1 . Introduction: To build up a brand two questions often arises: What makes a brand strong? And how do you build a strong brand? To help to find out we study about the CUBE model which has four steps or six blocks. Beside this marketer face two questions are: What o different brands mean to customer? And how does the brand knowledge of consumers affect their response to marketing activity? The challenge for marketers in building a strong brand is ensuring that customers have the right type of experiences with products and services and their accompanying marketing programs so that the desired thoughts, feelings, images, beliefs, perceptions, opinions, and so on become linked to the brand. 1. 2. Origin of the report: Brand is a very sensitive issue to the marketing people and is a very important issue to the customer. Because a strong brand can create a very highly demand of a reduce of a particular brand. To build up a strong brand one should to know so many things, ; he/she have to go with a four step procedures. And in this report we tried to focus on building a brand as a strong brand for that we select a local fashion brand. 1. 3. Objectives of the study: We make a study to know the Brand identity, meaning, response, relationship with the customers. We tried to know that is the customers are satisfied, dissatisfied, confused about the brand, its service, products and the overall factors. The core or the main objective of the study is to building a local fashion brand. To find the robbers, and scope of a local fashion brand ; build it as a strong brand. 1. 4. Methodology: We make a survey on Dishes Doss, which is a local fashion brand with ten famous local fashion brands. We make a questioner consisting 69 questions, on the basis of brand building in categories. Then take a sample of 50 people of different age, gender, ; occupation who use these brands. After getting the answers, we measured the level of satisfaction, dissatisfaction, confusion on different level of brand building. . 5. Limitations of the study: We are in our formal education stage. So this is our first assignment which is n the basis of the practical experience in real life. So, our lack of experience greatly influenced in preparing this report. We faced some other limitations that are given bellow: * It is too much difficult to comment and suggest based on few days study. Assignment report is import ant to fulfill the Brand Management course, but we do not have enough time for such a detailed process. * Lack of collecting essential data ; information. * The survey is not 100% accurate. The four steps of brand building: The four steps of brand building represent a set of questions that customers invariably ask about brands. The questions are as follows: 1. Who are you? (Brand identity) 2. What are you? (Brand meaning) 3. What about you? What do I think or feel about you? (Brand response) 4. What about you ; me? What kind of association and how much of a connection would I like to have with you? (Brand relationships) By establishing the answer of these questions marketer can build a strong brand in the market among the customers. When customers get a clear answer of their question from the marketer then it become easy to make a decision about the brand. When marketer able to establish he answer of the question then they are on the way of building a strong brand. 2. 1. 1 . Brand Identity/salience: Achieving the right brand identity meaner brand salience with the customers. Brand salience measure awareness of the brand, that indicate is the customer are aware of the brand or not? Or how often and how easily the brand is identified by the customer in order of various situations and circumstances? We have said that brand awareness refers to customer ability to recall and recognize the brand under different conditions and to link the brand name, logo, symbol, and so forth to certain association in memory. Here we will know about the depth and the breadth of awareness thus gives the product an identity by linking brand elements to a product category and associated purchase and consumption or usage situations. Brand depth measure that how likely or easily the brand element come to the mind of customers ; brand breadth measure the range of purchase ; usage situation in which the brand element come to the mind. . 1. 2. Brand Meaning/Performance ; Imagery: The brand performance describe how well the product or service meets customers’ more functional needs. How well does the brand rate on objective assessments of laity? To what extent does the brand satisfy utilitarian, aesthetic, and economic customer needs and wants in the product or service category? Five types of attribut es and benefits often underlie brand performance, as follows: 1. Primary ingredients and supplementary features. 2. Product reliability, durability, and serviceability. 3. Service effectiveness, efficiency, and empathy. 4. Style and design. 5. Price. 2. 1. 3. Brand Response/]judgment feelings: Brand image indicate a brand picture in a customer mind. Brand imagery depends on the extrinsic properties of the producer or service, including the ways in which the rand attempts to meet customer psychological or social needs. In this way people think about a brand abstractly, rather then what they think the brand actually does. Thus imagery refers to more intangible aspects of the brand, and consumers can form imagery association directly from their own experience or indirectly through some other source of information. Many kinds of intangibles can be linked to a brand, but four main ones are: 1. User profiles. 2. Purchase and usage situations. 3. Personality and values. 4. History, heritage, and experiences. Brand Judgments are customers’ personal opinions about and evaluations of the rand, which consumers form by putting together all the different brand performance and imagery associations. The four types of brand Judgments are important, they are: 1 . Judgment about brand quality, 2. Brand Credibility, 3. Brand Consideration, 4. Brand Superiority. 2. 1. 4. Brand Relationship/Resonance: Brand resonance describes the nature of the relationship and the extent to which customers feel that they are ‘in sync’ with the brands. Resonance is characterized in terms of intensity, or the depth of psychological bonds that customers have with the brands, as well as the level of activity engendered by this loyalty. How to cite Building a Local Fashion Brand, Essays

Marketing orientation concepts and management - MyAssignmenthelp.com

Question: Discuss about the Marketing orientation concepts and management. Answer: Corporate objectives Corporate objectives of Apple constitute of various aspects, which provides holistic scenario for all the associate stakeholders. Corporate strategy for Apple includes offering innovative and up-to-date products to their customers. This is one of their key objectives due to the reason that, Apple is known for their innovative and advanced products compared to their competitors in the market. Innovative products of Apple are one of their key competitive advantages in the market (Heracleous, 2013). Another corporate strategy of them is the enhancement of the value for their stakeholders including both internal and external stakeholders. This enables them to look after the interest of their stakeholders. Moreover, this corporate objective of them is related to the basic business concept of maximizing sales and revenue. This is due to the reason that, maximization of the sales and revenue will automatically enhance the value for the internal customers. On the other hand, proving quality and innovative products will also enhance the value for the external stakeholders. In addition, Apple also concentrates on providing aesthetic value to their customers in order to enhance their value (Cecere, Corrocher Battaglia, 2015). Thus, they are having universal approach in initiating their corporate strategy covering all the related aspects. Market orientation Marketing orientation is having three concepts namely, marketing concept, selling concept and production concept. Marketing concept is the most popular concept being followed by the contemporary business organizations (Urde, Baumgarth Merrilees, 2013). It refers to the concept of determining the taste, preference and requirement of the customers as well as of the whole market and offering products accordingly. Selling orientation refers to the concept of promoting and marketing the products being manufactured. In this case, the key concentration is being given on the promotional and selling activities for the particular product. Production orientation was being used till the late fifties. This refers to the concept or thinking that, producing good quality or high quality product will automatically have market or they will create market. In this case, preferences of the customers are not being determined. Apple follows both the product and market orientation. This is due to the reason that, Apple relies more on their quality of products, which creates standard and market of their own. Every time, Apple introduces upgraded versions of iphones, a new standard in the market of mobile device is being created (Kam Sing Wong Tong, 2012). Moreover, iphones are not being able to meet all the requirements of the customers but still they are popular in the market. On the other hand, they also follow marketing strategy, which helps them to determine the market requirement prior to the release of the products. Core marketing strategy The core marketing strategy of Apple comprise of various approaches including pricing, product quality, branding. The key marketing objective of Apple is to cater to the niche customer segment by providing quality and aesthetic products to the customers. Accordingly, they initiate premium pricing in order to target the high end customers and to enhance the exclusivity of the products. In addition, branding is being done in such a way that it will create premium image in the market and will attract the customers from the top of the pyramid. However, there are various strengths and weaknesses being identified in this strategy of Apple. One of the key strengths is the positive branding of the products. With the help of premium pricing and marketing strategies, higher end customers are being effectively catered, which shows in their sales chart (Banerjee Soberman, 2013). One of the key weaknesses of this strategy is the non-coverage of the entire market. Though Apple is one of the renowned and sought after brand in the market, but they do not offer mass market and affordable products, which refrain them from generating more volume from the market. As discussed earlier, corporate objective of Apple is to provide quality products and maximizing revenue. Accordingly, the marketing strategies of them including initiation of premium pricing and innovation help them to generate more revenue and offering good quality products. Reference Banerjee, S., Soberman, D. A. (2013). Product development capability and marketing strategy for new durable products.International Journal of Research in Marketing,30(3), 276-291. Cecere, G., Corrocher, N., Battaglia, R. D. (2015). Innovation and competition in the smartphone industry: Is there a dominant design?.Telecommunications Policy,39(3), 162-175. Heracleous, L. (2013). Quantum strategy at Apple Inc.Organizational Dynamics,42(2), 92-99. Kam Sing Wong, S., Tong, C. (2012). The influence of market orientation on new product success.European Journal of Innovation Management,15(1), 99-121. Urde, M., Baumgarth, C., Merrilees, B. (2013). Brand orientation and market orientationFrom alternatives to synergy.Journal of Business Research,66(1), 13-20.

A Personal Evolution free essay sample

The question: Comment on the changes that have occurred in your life during the last four years? I was recently fortunate enough to conduct an interview with bonafide movie star Colin Farrell. Most of our discourse pertained to film, his upcoming projects, and other cinematic ambitions. However, towards the end of our conversation he poignantly observed that when he first started making movies in the states, he was terrified of the prospect of going back to his hometown in Ireland and having his friends and family proclaiming â€Å"he’d changed†. Before I could contemplate how I’d evolved over my miniscule 18-year life, Ferrell followed that sentiment with â€Å"but you should be changing all the time.† Farrell’s final words had ignited a notion I’d been ruminating for the past few years: if there’s one constant in this actively evolving world, it’s the act of change. Colin Ferrell had experienced it, my family has experienced it, and now I have experienced it. We will write a custom essay sample on A Personal Evolution or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page In the past four years there have been several events – some monumental, some existential, and some seemingly inconsequential that have shaped and altered my life. Chronologically (and perhaps on a scale of severity) the first substantial change occurred when my mother packed up, took my two brothers, and relocated to California. I stayed behind with my father in Chicago. My decision was met with equal amounts of anger, trepidation, sadness, and angst from my endearing mother (whom I regularly lived with up until that Freshman year). Those ten months spent living with my father and attending a rigorous Catholic private school weren’t the most pleasant of times. To be more specific, my freshman year of high school was a series of dismal events filled with mean-spirited classmates, lackluster teachers, and a disastrously counter-productive addiction to gaming on my front. It’s a part of my history that I don’t tend to reflect on – and in no part was my father responsible for the awfulness of that year. In fact, he may have been the only redeeming feature in those times. Alas, I evolved and decided to move out west with my mother and two brothers. This was a time for me to reinvent myself – to forget about the past year and start fresh with a clean slate. Moving to California provoked an infinite amount of change in my life. It began with my focus shifting from excessively indulging in electronics to writing about the art form of film. Film directly assisted with my growth: it opened up a new world for me spend endless hours in, ideas to contemplate for days on end, and stories that enlighten my mind and brought tranquility to a life that was often occupied with stress, drama, and heartbreak. As my career in film criticism evolved (I was accepted as a member of the press in California, attended film festivals around the country, and obtained several jobs as a reporter), so did my life at home. I was no longer the perpetually ambivalent and disgruntle teenager my parents could hardly tolerate. The days of isolation came to an end as well. Friends were made, academics took priority, and happiness took precedence. It wasn’t all peaches and roses, but as time went by I began to realize just how fortunate I was to be living. My appreciation of family, friends, and love overtook my preconceived cynical notions of life. And so, I revert back to Colin Farrells philosophy on change in our lives. While evolving isn’t always beneficial, it is natural. It’s comforting to know where I stand in my life as this moment. By the same token it’s exciting to contemplate where I’ll be in four years – and who and what in that time will change who I am. Only time will tell.

Cuánto demoran los papeles para esposos de residentes

Cunto demoran los papeles para esposos de residentes La peticià ³n de papeles para su cà ³nyuge realizada por un residente permanente se demora, en el momento de escribir este artà ­culo, entre aproximadamente 21 meses de mà ­nimo y 30 de mximo. En este artà ­culo se explican las causas de la demora, cà ³mo es posible hacer un clculo ms detallado, segà ºn el caso de cada solicitante, cosas que deben saberse mientras se estn en el medio del proceso de la tramitacià ³n y, por à ºltimo, quà © causas pueden dar lugar a la negacià ³n de peticià ³n de la tarjeta de residencia, tambià ©n conocida como green card. 4 factores que determinan la  demora la green card para cà ³nyuges de residentes La demora de la peticià ³n, la cual se puede presentar para un cà ³nyuge del mismo sexo o del sexo opuesto,  depende, bsicamente, de cuatro: En primer lugar, de quà © oficina del Servicio de Inmigracià ³n y Ciudadanà ­a (USCIS) debe aprobar la peticià ³n En segundo lugar, de la oficina consular en la que se realiza la entrevista para los casos de cà ³nyuge fuera de Estados Unidos o de la oficina del USCIS que debe aprobar el ajuste de estatus, para el caso de cà ³nyuges en situacià ³n migratoria legal en los Estados Unidos En tercer lugar, del nà ºmero total de peticiones de esta categorà ­a, ya que existe un nà ºmero mximo que se puede aprobar por aà ±o fiscal y asà ­, cuando se alcanza dicho nà ºmero las oficinas del gobierno paralizar el nà ºmero de peticiones aprobadas hasta que el inicio del siguiente aà ±o fiscal. Y en cuarto lugar, del paà ­s de nacimiento de la persona pedida ya que para Mà ©xico aplican unas fechas y para el resto de Latinoamà ©rica y Espaà ±a, otras.   Pero es posible calcular las demoras y tambià ©n verificar cada poco si hay cambios en los tiempos de tramitacià ³n. Adems, si todavà ­a no se ha iniciado el trmite, esto es lo que se debe  hacer para que un residente solicite la green card para su cà ³nyuge. Paso 1: demora el USCIS en aprobar la peticià ³n El primer paso en la obtencià ³n de la green card para el cà ³nyuge es que el USCIS apruebe la solicitud. El paquete con ella se envà ­a a una oficina (depende del lugar en el que se reside) y, una vez, allà ­, se distribuye entre los llamados Centros de Servicio. A las pocas semanas de haber enviado el paquete con la solicitud se recibe una carta que tambià ©n se conoce con el nombre de NOA1. Contiene informacià ³n importante, como el nà ºmero de caso y tambià ©n el lugar en el que va a tramitar la solicitud.   Una vez que se sabe cul ha tocado, ir a esta pgina oficial, bajar el cursor, elegir el Centro de Servicio adecuado y presionar en centro de servicio fechas de trmite.   A partir de ahà ­ se abre una pgina y hacia la mitad de la misma buscar formulario I-130 peticià ³n para pariente extranjero y verificar la opcià ³n de peticià ³n de un residente permanente para un cà ³nyuge o un nià ±o menor de 21 aà ±os.   Y el tiempo de procesamiento son los meses en los que el USCIS se demora para contestar si aprueba la peticià ³n o la deniega. Lo notifica con una carta conocida como NOA2 que es muy importante porque ahà ­ aparece la fecha de prioridad. Paso 2. La gran espera por los papeles Una vez que el USCIS aprueba la solicitud hay que esperar por una visa disponible. Esto quiere decir que por ley hay un nà ºmero limitado de peticiones de tarjetas de residencia que se pueden hacer en esta categorà ­a de solicitudes de residente para esposo, lo que se conoce como F2A.   Y como el nà ºmero de solicitudes es ms grande que el nà ºmero de visas disponibles entonces se producen unos retrasos.   Cada mes el Departamento de estado publica el boletà ­n de visas. Ahà ­ es el lugar donde informarse  si ya hay visa disponible y se acabà ³ la espera. La manera de mirarlo es teniendo la fecha de prioridad en la mano. Si la fecha es anterior al dà ­a que el boletà ­n de visas seà ±ala para la categorà ­a F2A, eso quiere decir que comienza de nuevo la tramitacià ³n que falta.Si es ms reciente, hay que seguir esperando. En la actualidad, el boletà ­n de visas est dividido en dos grandes categorà ­as.   En el caso en que se puede ajustar el estatus, verificar la fecha en dates for filing. Por el contrario, los casos con procedimiento consular deben consular el final action. Paso 3. Ajuste de estatus o procedimiento consular Entonces pueden pasar dos cosas segà ºn dà ³nde se encuentra el cà ³nyuge pedido: Si est en Estados Unidos legalmente se puede proceder a su ajuste de estatus. Y aquà ­ se explica cà ³mo verificar los tiempos de demora para eso. Recordar que los esposos de residentes en situacià ³n migratoria de ilegalidad no pueden ajustar su estatus y, por lo tanto, antes de iniciar cualquier trmite migratorio deberà ­an consultar y asesorarse con un abogado. Si la persona pedida est fuera de Estados Unidos habr unas gestiones (como envà ­o del affidavit of support) con el Centro Nacional de Visas y los pasos finales se harn en el paà ­s de cada uno, incluyendo una entrevista en el consulado o Embajada. Aunque las demoras varà ­an por paà ­s en là ­neas generales puede decirse que desde que hay visa disponible a la finalizacià ³n del proceso no pasan ms de seis meses.   Problemas en la peticià ³n de la green card para esposos Estas son 42 causas por las que se deniega la green card. No todos los problemas son iguales, ya que en algunos casos es posible pedir un perdà ³n, tambià ©n conocido como waiver. En otros casos, el castigo es solamente temporal. Finalmente, en algunas circunstancias no ser posible encontrar arreglo y no se obtendr la green card. Adems, tratndose de un matrimonio, el USCIS y, en su caso, el consulado va a poner atencià ³n para averiguar si el matrimonio es real o es sà ³lo de conveniencia.   Recordar que los  esposos de residentes que estn en Estados Unidos como indocumentados  no pueden ajustar su estatus por matrimonio. Con lo que si los piden en esa situacià ³n tendrn que salir de Estados Unidos y allà ­ puede saltar el problema del  castigo de los 3 y de los 10 aà ±os. Por todas estas razones dichas, es recomendable antes de iniciar trmite asesorarse con un buen abogado de inmigracià ³n cuando alguno de esos problemas puedan aparecer. Mientras se est esperando por los papeles Mientras se espera por la green card, puede suceder que el solicitante cambie de estatus legal al naturalizarse y convertirse en ciudadano estadounidense. Esto es lo que debe  hacer si el residente se convierte en ciudadano.   Por otra parte, tambià ©n mientras se espera por la green card si el cà ³nyuge est fuera de Estados Unidos puede que desee viajar para reunirse con su esposa o marido. Pero esto no siempre es posible y conviene enterarse de las posibilidades de ser exitoso al  pedir una visa de turista mientras se espera por green card. Asimismo, como las cosas de amor pueden ser cambiantes, es importante conocer cà ³mo  afecta el divorcio a la green card. Consejos para que todo el trmite resulte bien Estados Unidos es un paà ­s diferente en muchos aspectos como por ejemplo, el migratorio, el legal, o en materia de impuestos, etc. Para evitar problemas y conocer los derechos todas las personas que desean emigrar deberà ­an familiarizarse cuanto antes con los aspectos bsicos del paà ­s y asà ­ evitar ms tarde problemas indeseados.   Adems, este es un listado de telà ©fonos y pginas webs en los que se puede verificar informacià ³n sobre el caso y obtener buenos recursos migratorios.   Finalmente, se aconseja tomar este quiz - trivial o test- para verificar que tienes los conocimientos bsicos para obtener y conservar la tarjeta de residencia. Es difà ­cil conseguirla. No corras el riesgo de que te la cancelen por ignorancia. Este artà ­culo es sà ³lo informativo, no es consejo legal.

Appeal Process Essay Example | Topics and Well Written Essays - 500 words

Appeal Process - Essay Example er of cases although some may require to be heard by separate appellate committees such as the appellate committee of bankrupt judges (Lawfirms.com, n.d.). A litigant wishing to appeal does it only on paper, and at the appeal, level there is no actual trial. The litigant will have to write down a brief explaining how the court erred in his sentencing, or how their constitutional rights may have been violated in the trail. The federal or state cause trying this case will review the brief and determine based on the submitted evidence if it is a legitimate cause for a retrial. In some cases, the appeal courts will require having an oral argument by the lawyers; this does not involve submission of any new evidence (Lawfirms.com, n.d.). While the appellate court has the legal mandate to review any facts pertaining to the trial, the review process is based on what was on record on the court. It can review the facts of the case, but cannot overturn decisions on factual grounds without categorical evidence of erroneous decisions. Court decisions are not infallible since they are made by humans who are fallible and as such, it is crucial that people are given as the benefit of doubt for as long as possible and all loose ends tied up, this can only be achieved by an improved appeal process. Appeals are often long and tiresome processes, which have an incredible toll on the plaintiff and their families; one of the main causes for this is the shortage of appeal court judges. For an appeal to go through it must be heard by a panel of judges, and this is difficult if there are not enough to go round, another way of simplifying the appeal would be to make it possible for non-lawyers to appeal cases when they are not able to afford and attorney. This could be made possible by simplifying the legal and other requirements needed in the cause of pursuing the appeal. Antony William was arrested and arraigned in for the murder of a ten-year-old girl, while being transported to

Issues in sports 1 Essay Example | Topics and Well Written Essays - 500 words

Issues in sports 1 - Essay Example The women players or athletics are under many constraints related to economical, financial, social and political issues. Most of the conservative countries are not permitting women to participate in any sports event. The social and political rules and regulations are against the women. Some countries are not providing enough financial assistance to the women team or women athletics but they are very much interested in putting more money for the men’s team or athletics. According to Kirsch, â€Å"In todays world female athletes are learning the hard way that even if they train harder than their male counterparts and out-perform them they will still not receive the same recognition or money that the guys make†. (Kirsch) Recently, some countries are giving importance to their women players and athletics. United Nations, China and some other developed and developing countries are trying to remove this discrimination through proper implementation of new policies, which help women players to come out from those constraints. The United Nations Millennium Development Goals (MDGs) introduced innovative ideas to solve this discrimination problem. This was a great achievement for women athletics and players. It was implemented to create gender equality in sports. In 2002, the United Nations Inter-Agency Task Force on Sport for Development took initiatives to give more importance on this discrimination issue in sports. They suggested some steps by which they can create gender equality. The steps are as followed, Drug abuse by athletes and sports persons is an international issue in sports. For instance East German succeeded in sports during the 1970s and 80s fighting back the United States and even Soviet Union with the help of intake of several performance enhancing medicines. This was meant to prove their power in sports against the west. Swimmers like Kornelia Ender, Barbara Krause and Carol Mitschke were the victims of the doping schemes. During 1989

Compare the relative qualities and characteristics of Modular Essay

Compare the relative qualities and characteristics of Modular Information Systems against Integrated Software Systems in respect - Essay Example This information is produced to meet a business objective and it is valuable to the company. Information system is a set of elements which collect data, process and give the output data and information (Sommerville 2010). Business information systems are used by businesses and organizations for storing, processing and analysing data. The information systems are useful to the organizations since they enable the employees to recognize new patterns and relationships in the stored data. This enables the companies to make important decisions which ensure that the organization gains a competitive advantage in the market. The information systems ensure that the organizations activities are conducted efficiently and reliably and this improves the overall performance of the business. There are different types of information systems and this leads to variations in the functionalities of business systems. Computer systems are software programs that are developed to capture, store and manipulate data. There are different types of computer systems in the market and this depends mostly on the functionality of the software (Ajmani 2004). There are different processes of developing software programs and this result to different software products with different characteristics. The process of developing that system is crucial to the functionality of the systems. Modular information systems are comprised of different modules that are connected together. The development of this system is done by partitioning the overall system into distinct parts known as modules. The modules have different functionalities and are developed independently. This type of development is preferred by organizations which have different functionalities and require detailed processes in the software systems. The modules are then integrated to form one huge system which can be used by an organization to conduct its business activities. Integrated software systems are developed as single units and the func tionality of the system is achieved by the whole system. Most of the integrated software systems are off-the shelf systems which are bought directly from the software dealers (Bass,et al., 2008). The functionality of the system meets the requirements of the organization and there is no need for gathering specific requirements of the organization. Modular information systems are flexible. This is because it is possible to replace or add one component to the system without affecting the other rest of the system. This is useful in organizations that are involved in many activities which are dynamic (Clements and Kazman 2008). The new activities that are carried out by an organization might require the development of a new system to meet the functionality required by the organization. Organizations which have modular systems can easily integrate new modules to meet their requirements. This is effective as there is little interruption to the business activities since one module is only a ffected. Integrated systems might result to the disruption of the whole system as new functionalities to the existing systems cannot be added without affecting all the system processes. The modular information systems perform better than integrated systems. The modular systems are developed from requirements that have been gathered from the organization. This ensures that the system is developed to perform better in the

Analysing The Challenges Facing Multilateral Organisations Today Politics Essay

Analysing The Challenges Facing Multilateral Organisations Today Politics Essay War is often the result of non negotiable problems/disputes. Since the end of the Second World War, liberal international theory advanced the establishment of international organizations such as the United Nations (UN) and its sub bodies, in the belief that such international forums were where state members have a chance to discuss and air their common issues and grievances and could prevent war and, widen the corporations between nation states. However, since its creation, the legitimacy as well as the effectiveness of these multilateral institutions has always been questioned. The end of the Cold War marked a new era of world politics where the United States (US) became the predominant power in the international system. In addition, the Iraq war of 2003, led by the US, but was not authorised by the United Nations Security Council (UNSC) and the issue raised a question about the hegemonic position of the US over the United Nations. This is just one of the many challenges multilatera l organizations like the UN are facing today. The terrorist attack on 9th November on the US and similar attacks elsewhere in the world, as well as the increasing threat of weapon of mass destruction (being used by either a rogue state such as Iran or a terrorist group) in the 21st Century is another major challenge for the United Nations and its administrative departments. Moreover, with the increasing involvement of non state actors and non governmental organizations nowadays, the value of formal institutions like the UN is clearly affected in adverse ways. To clarify if multilateralism is in crisis or not, it is essential to firstly look at the difficulties posed by these new dimensions to multilateralism, then evaluate its reaction and effectiveness at tackling them. In this essay, the focus will be on UN as this is often seen as the vanguard of multilateralism. Firstly, the essay will examine what multilateralism can be defined as and will name the current challenges which Mult ilateralism faces, it will then go on to give analysis of the sources of those troubles. Finally, it concludes that such multilateral institutions like the United Nations are under real challenge to the effectiveness of their multilateralism; however, it is not yet in a crisis situation. The leaders of these organizations can no long presume their exchanges to be business as usual but need to take into account these growing troubles and think more about reform if multilateralism is to continue to function in a stable way. Before discussing whether multilateralism is effective one needs to consider what exactly multilateralism is. For Paul Nielsson, UN Commissioner, multilateralism is about all parties carrying out concerted efforts to strengthen the international regulatory framework (Jorgensen, 2007, p.2) and involves states pooling both resources and sovereignty. The problem for many participants in multilateral processes is that the processes themselves are often slow and difficult, and rarely do their results satisfy every participant (ibid). Thus, multilateralism is by definition a process where participants cannot expect a zero-sum outcome, where they can clearly identify their gains or losses and say it was a success or failure. Multilateralism is a compromise between states where its greatest success is that everyone is a winner and everyone is a loser, because nobody gets everything they want. However, the UN has become the focus of most peoples idea of what a true multilateral organisation is and should be. Therefore the success of the UN in resolving modern international problems is also used as the guideline to whether or not multilateralism is effective or ineffective. The recent invasion of Iraq led by the Bush administration was preceded by concerted attempts to make it a UN operation. The fact that it did not led to US claims that the UN was ineffective and prior to the invasion of Afghanistan the US president Bush said the UN was so ineffective that dont call us, well call you (ibid). However, the debate between whether unilateralism or multilateralism is the most effective method of international relations is one fought as much within America as it is internationally and as Nye observed This battle between multilateralism and unilateralism is often played out between the president and Congress and has led to a schizophrenic America (Nye, 2000, 156). A final problem is that the UN always focuses on promoting free trade as a vehicle for expanding multilateralism and economic power is focused still within the major developed Western states. Thus, developing states often see the UN as biased and ineffective when it comes to promoting the interests of the Third World (Jorgensen, 2007, p.4) However, despite the criticism for the lack of UN support for Iraq and Afghanistan invasions and thus the claims by many unilateralists that it is an international body that is largely ineffective, there remain many who still see multilateralism as the finest success of the post- war environment. The Deputy Scretary-general Asha-Rose Migiro of the UN made it clear in a speech in 2009 that the world faced more than problems of security and terrorism. These include poverty and hunger in less developed countries, the lack of protection in many areas of human rights, the global financial crisis and discrimination against women. For Migiro these problems can only be solved through, global solutions to global problems and the UN is the only existing global organisation that can deal adequately with these challenges (Migiro, 2009, p.1). Other multilateral organisations exist but in Migiros view Solutions must be hammered out in a process involving all States.   Groups such as the G-8 and G-20 are important, but it is vital to return to the United Nations as the natural locus of action on global issues (Migiro Asha-Rose , 2009, p.2). War is often the evidence of a failure of multilateralism and the UNs avoidance of actually getting involved in the wars in Iraq and Afghanistan maybe seen as an effective decision rather than a failure, especially as the UN is still often the first organization that enters into a conflict arena during and after the conflict in order to either make peace or sustain a peace process. The Secretary-General made it clear that there was no choice available between multilateralism and unilateralism as We either succeed together, or we fail alone (ibid). Obviously, the avoidance of involvement in conflict is often used by participants in the conflict as a reason why the UN and multilateralism is ineffective. However, the response to aggression in international affairs is often best tackled through multilateralism. AsKevin Hartigan argues, multilateralism is a demanding organisational form. It requires its participants to renounce temporary advantage and the temptation to define their intere sts narrowly in terms of national interests, and it also requires them to forego ad hoc coalitions and to avoid policies based on situational exigencies (Hartigan, 1992, p.604). The UN has proven to be the most effective forum within which this cooperation can occur, as witnessed in the first Gulf War. In defence of the UN, Shashi Tharoor, former UN Under-secretary General noted that relative peace in the second half of the last century depended on the idea that in order to keep the peace, in order to help human beings to progress and so on, you needed a mechanism, as well as a system of rules that would actually be to the benefit of all and that this role could only effectively be carried out by the UN (Tharoor, et al, 2006. P.4). As Shashi notes, todays world is one with problems without passports (such as terrorism, climate change, drug trafficking) and the UN provides a forum for states to discuss and develop solutions to these problems on an international level of cooperation. Tharoor later observes that the problems of today are often too large, too complex for one government, or bi-lateral relationship to adequately deal with and so the involvement of the UN means, that humanity is responsible, not one government.   In that process, the universality of the United Nations gives you a mechanism to actually deliver effective results. On the first hand, the value of those collective institutions like WTO or UN is undeniable, especially in the time of globalization when the world is becoming a borderless entity where not only trade, but also problems such as drugs and climate change cross from country to country unimpeded by the sovereign idea of borders between states. The UN reduces the expense, time and effort needed by any one country to deal with threes problems. It offers a forum within which states can confidentiality work out deals and strategies, and in the process states get to know about each other freely, thus increasing the confidence in future levels of cooperation (Robert o Keohen, p2). Furthermore, as international terrorism and transnational criminals are increasing problems then it bis easier to see that a threat to one is a threat to all. Very few states have the power to stand alone and confront those difficulties ( A more secure world, 2004). Although UN has been criticised for not responding e ffectively to these emerging challenges, there is little in the way of alternatives being offered. As Robert argued; terrorism and proliferation of weapons of mass destruction can only be addressed in concert with other countries, even the super power like US(Hutchings) does not have sufficient resources, political will or power to deal with these modern challenges. However, the UN is aware that it needs to reform and has set out five primary areas for change. Reform may involve drastic changes to the members and size of the Security Council and to the way it makes decisions, but essentially it needs reforms that assist it in, 1. Reforming management and operation procedures, 2. Strengthen humanitarian action, 3. Bring human rights to all in the world, 4. Strengthen efforts to maintain peace and security, and, 5. Significant movement forward on meeting development challenges (UN). Thus the extent of the need to reform to meet these challenges goes beyond just altering the size and methods of working of the Security Council. It requires dealing with the financing of the organisation, strengthening aspects such as the Human Rights Council and ensuring that the popular promotion of multilateralism is always more effective and long-lasting solutions to the modern problems of the international system rather than the unilateralism so often chosen by s tates and directly the cause of many of the wars of today and in the past. Hartigan, Kevin (1992), Matching Humanitarian Norms with Cold, Hard Interests: The Making of Refugee Policies in Mexico and Honduras, 1980-1989, International Organisation 46, Summer, 604. Knud Erik Jà ¸rgensen(2007) The European Unions International Identity: the Role of Multilateralism Nye, Joseph (1991) Bound to Lead: The Changing Nature of American Power (Basic Books 1991). Nye, Joseph (2002) The Paradox of American Power, Ikenberry, G. John (2002) Americas Imperial Ambition, Foreign Affairs, 81(5): 44-60. Migiro Asha-Rose (2009) Tackling Current Global Challenges Requires Building An Effective Multilateralism, Speech to UN, 15th April 2009 Shasis Tharoor, et al (2006) DebateThe United Nations: Still Relevant After All These Years? Carnegie Council Essay question : Is multilateralism under challenge? Or in crisis? Or is it business as Questions to define What is multilateralism, the definition, are we talking specifically about UN or any other? IR theory, Realist, Liberalistà ¢Ã¢â€š ¬Ã‚ ¦ Crisis? How do you value /define/ classify if M is in crisis? Or is it only about emerging challenges?( in that case, list the challenges), in 21st century The increasing development/involvement of non-state factors not necessarily means the fail of formal institutions. Ad hoc coalition. Alternative arrangements. Strength and weakness. Argument: challenges are real. Ineffectiveness is real. Neither takes it as crisis nor business as usual. Need to fix it. Need to reform. Ref Robert L. Hutchings, 2003, The United Nations and the Crisis of Multilateralism, Keynote Address, University of Pennsylvania, Model United Nations Conference http://www.dni.gov/nic/speeches_un_multilateralism.html Keohane, Robert O., 2006, The contingent legitimacy of multilateralism, in Edward Newman, Ramesh Thakur and John Tirman eds., Multilateralism Under Challenge? Power, International Order, and Structural Change (Tokyo: United Nations University Press). Edward Newman, Ramesh Thakur and John Tirman eds., Multilateralism Under Challenge? Power, International Order, and Structural Change (Tokyo: United Nations University Press), introduction. Newman, Edward, 2007, A Crisis of Global Institutions? Multilateralism and International Security (Oxford, Routledge) Introduction, chapter 1 and 2. A More Secure World: Our Shared Responsibility, Report of the Secretary-Generalà ¢Ã¢â€š ¬Ã… ¸s High Level Panel on Threats, Challenges and Change (New York: United Nations, 2004), summary. Available at: www.un.org/secureworld Forman, Shepard, and Derk Segaar, 2006, New Coalitions for Global Governance: The Changing Dynamics of Multilateralism, Global Governance, vol.12, no.2.

Does Vision And Mission Emerge Essay -- essays research papers fc

Does vision and mission emerge from the particular culture of a firm or is it dictated by strategy?   Ã‚  Ã‚  Ã‚  Ã‚  The culture of a firm and the formation of strategy are two very important aspects of an organisation. The world contains many different people all with different values, ideas and beliefs. These differences create a diverse range of cultures within organisations, some having bigger influences than others. Strategies within organisations are highly dynamic and complex, and can have positive and negative effects on an organisation. Vision and mission are concepts that many believe are vital for an organisation to operate effectively and to the best of its abilities. Andrew Campbell (1991) , a prominent writer on vision and mission, believes that both the culture and the strategy of a firm come together side by side to build an overall definition of mission for a firm. The paper will be based around Campbell’s perception and whether either culture or strategy has a greater part to play in an organisation’s mission.   Ã‚  Ã‚  Ã‚  Ã‚  Culture and strategy will be examined in a context relevant to the title question. Hofstede (1993) defines culture as ‘the collective programming of the mind which distinguishes one group or category of people from another’. Hofstedes research of different organisations and countries allowed him to make predictions on the way different societies operate, including their management processes and the theories that would be used in management. Morgan (1996) refers to culture as ‘the pattern of development reflected in a society’s system of knowledge, ideology, values, laws, and day-to-day ritual’. In subsequent writings he expands on organisational culture as ‘self-organising and is always evolving’ and also ‘ we are observing an evolved form of social practice that has been influenced by many complex interactions between people, events, situations, actions, and general circumstances’. These broad definitions of o rganisational culture are important bases and will be used throughout.   Ã‚  Ã‚  Ã‚  Ã‚  Defining strategy is a difficult process because it is a complex concept that has many forms and is constantly changing. Andrews (1987) attempt is a comprehensive definition... ...(1980) Defining the Business – The Starting Point of Strategic Planning New Jersey, Prentice Hall Collins, J.C. & Porras, J.I. (1996) Built to Last – The Successful Habits of Visionary Companies Kent, Century Ltd. De Wit, B. & Meyer, R. (1998) Strategy – Process, Content, Context London, International Thomson Business Press Fawn, J. & Cox, B. (1985) Corporate Planning in Practice London, Kogan Page Ltd. Morgan, G. (1996) Images of Organizations London, Sage Publications Porter, M.E. (1985) Competitive Advantage New York, The Free Press Journals Baetz, M.C. & Bart, C.K. (1996) Developing Mission Statements Which Work Long Range Planning 29 (4), pp.526-533 Campbell, A. & Yeung, S (1991) Creating a Sense of Mission Long Range Planning August pp.10-20 Campbell, A. (1997) Mission Statements Long Range Planning 30 (6), pp.931-932 David, F.R. (1989) How Companies Define Their Mission Long Range Planning 22 (1),pp.90-97 Piercy, N.F. (1994) Mission Analysis: An operational approach Journal of General Management 19 (3), pp.1-19 Hofstede, G (1993) Cultural Constraints in Management Theories Academy of Management Excutive 7 (1)

History Essay

Essay Question: Underlying causes of 1905 revolution in Russia were not political. Whatever else the revolutionaries wanted it was not to overthrow the Tsar. How far do you agree with this claim? I agree that the underlying causes of the 1905 revolution were not only political there were also economic, social and military factors that lead to this revolution. Firstly the social causes of this event. Even though in 1861 the Emancipation of the Serfs had occurred by the early 20th century nothing really had changed for the lower class of Russia society. They were not called Serfs anymore and no longer were a part of a feudal system however the still experienced a low standard of living, had to work long hard hours in menial jobs and poor harvests and famine havocked their lives. The massive gap between the rich and the poor continued to widen as the Tsar and the royal family lived in extravagant luxury and the peasants that made up 85 percent of the people were left to starve and fend for themselves. Another social factor than increased the peasants growing discontent with their ruler was the process of urbanization that led to overcrowding in the cities and horrible living conditions. Overall all these factors and more led to the popular belief among the Russian peasantry that change was needed, that change didn’t need to be the removal of the Tsar. In fact they were happy to work with him to achieve more equality and better living and working conditions for themselves. Next there were the economic causes of this revolution. In the 1900s the world was struck by a recession, Russia in particular was heavily affected. They were so affected because Russia relies so much on its exports especially of grain for its revenue and strength of its economy. So when the worldwide price of grain dropped during the recession mass unemployment results due to the fact that 85% of Russia populations are farmers that grow grain and other foodstuffs. Moreover due the recession the value of the Russia currency (the ruble) drops and therefore Russia exports of grain drops due to farmers not wanting to sell their grain due to an unstable market price. Widespread poverty among the masses grows leading to even more discontent and want for change. This leads on to my next point. The military cause of the revolution is simple and that is the Russo-Japan War. Tsar Nicholas II fearing the growing discontent among the people. So he comes up with an idea of a distraction to make the peasants forget about the horrible situation they are facing. At first his plan works well and the people forget about their troubles and support the war effort. That is until Russia starts losing. This idea needs to be put into context as Russia is the largest country in the world and should simply steamroll a tiny insignificant country like Japan. Defeats on land and at sea shocked the Russian public and if that wasn’t bad enough the war instead of making people forget about their struggles with poverty and starvation causes shortages of fuel and food, high prices and unemployment. As Russia suffered defeat after defeat they were forced to sign a treaty with Japan and became the laughing stock of European powers such as Germany and Great Britain. This shocking defeat also forced the Russia people to consider how incompetent the Tsar and his ruling party were. That they can’t even defeat a tiny island nation like Japan. However there were also political causes to the 1905 revolution such as the harsh Tsarist regime and the three main groups of opposition to the Tsar reign. The Tsar had already shown himself to be weak and indecisive through his actions again the Japanese in the Russo-Japanese war form 1900-1904. He had also made his views clear on the idea of a democratic/constitutional government calling them ‘senseless dreams’ and basically ignoring the pleas of his people for basic reform. The Social Revolutionaries tried to create a peasant revolution based around socialism and were a real ‘party for the youth’ but failed epically. The liberals appealed to the Tsar in a reforming manner but as I previously stated the Tsar refused to back down and give up any of his powers. However political causes amounted a minor role in the 1905 revolution as the other above factors were more important. Overall the underlying causes of the 1905 were not really political and while there was some political influence to the revolution. There were social factors such as urbanization and poor living standards. Furthermore economic factors such as poverty and unemployment ultimately led to dissatisfaction with the ruling class and change was required by the peasants. What the revolutionaries wanted was basic freedom e. g. freedom to form political parties, basic rights as human beings and fundament government reforms. Their main aim was for the people to have an interactive government system that sought the best for Russia. While it was true the peasantry of Russia was tired of autocratic rule they didn’t really want to overthrow the Tsar. All they really desired were some basic reforms to improve their lives.

The current law of Duress and Undue influence is so unclear that it creates chaos rather than certainty in the law Essay Example

The current law of Duress and Undue influence is so unclear that it creates chaos rather than certainty in the law Essay Example The current law of Duress and Undue influence is so unclear that it creates chaos rather than certainty in the law Paper The current law of Duress and Undue influence is so unclear that it creates chaos rather than certainty in the law Paper Essay Topic: Law If a contract is entered into under duress or undue influence this could be used to make the contract defective. However duress and undue influence are only normally allowed in exceptional cases. For example if a person knows the terms of the contract they are entering into, they enter into the contract with their eyes open and receive independent advise than they wont be able to get out of the contract. However the law on duress and undue influence is not this simple, as there seems to be little or no certainty in this area of law, the legal principles which have been established seem vague and many similar cases have conflicting decisions as the courts seem to apply the law flexibly. Through explaining this area of law illustrating the principles in the cases it will show how chaotic this area of law really is. Firstly there are two types of duress, physical duress that is where someone is forced into the contract against their will. The other form of duress is economic duress this is when one party exerts pressure on the other party to change the terms of the contract or renegotiate. However a party does have the right to alter the terms as long as there is consideration. However many cases where there is duress its in large commercial contracts and here its often difficult to make a distinction between hard bargaining and duress. Although this has to be established as hard bargaining is lawful however duress is not. The problem mainly with duress is that there seems to be little principles as, each case depends on its own facts, it must be shown that the victim had been coerced? When looking at the cases to help establish if the victim was forced into the contract it may need to be asked was it legitimate or illegitimate, lawful or unlawful, and did the victim obtain legal advise. However the cases dont seem to be consistent on this subject, especially when you compare similar cases. Examples of this include, Pau On v Lau Yiu Long [1980] AC 614 this case involved the sale of a building in Hong Kong, between the sale and completion of the contract there was a chance the shares would fall, buyer therefore said that the deal would be off unless other party made up difference if shares did fall. They did fall, the buyer than tried to use duress. The Privy Council decided the contract wasnt induced by duress, for duress it needs to be shown that it wasnt a voluntary act. So therefore in this case it was just hard bargaining. However in North Ocean Shipping Co v Hyundi Construction Co The Atlantic Baron [1979] QB 705, it was decided duress could be used, in this case the defendant owned a shipyard and had agreed to build the claimant a tanker for a certain price. However it was devalued by 10% so the defendant asked for 10% more money to make up the difference. And if he didnt pay this extra money they wouldnt build the ship or the next ship on order (which were both already rented out as soon as they were built. Duress was allowed to be used to void the contract. This case also said you should look at a persons conduct from the view of a reasonable man. These two cases demonstrate an inconsistency, why was Pau On v Lau Yiu Long found not to be duress, and The Atlantic Baron allowed using duress, they both have very similar circumstances. Maybe in the Atlantic Baron duress was allowed because the ships were already rented out as soon as they were made, so maybe there is more pressure here so its an involuntary act. Or maybe this just shows how inconsistent this area of law is, because although duress seems to be decided on the individual facts of the case, why have these two cases been decided differently with such similar facts. The case of The Universe Sentinel, Universe Tankship Inc v ITWF [1982] 2 QB 705, duress was allowed to be used as it was shown there was pressure amounted to compulsion of will and there was a illegitimacy of pressure exerted. Little of the cases on duress follow other cases as many of the cases it does not seem to be easy to tell whether its duress or not, as many cases could go either way. Even the judges are often uncertain; this is why there are no firm legal principles on this subject. Some examples of this are, CTN Cash Carry v Gallagher Ltd [1994] 4 All ER 714, in this case the defendant sold cigarettes at cut prices, if didnt pay more money to the other party they said they would end the contract and they knew the other party business were dependent on them. They tried to recover money under duress, there was no duress and it was said that, only because the business is in a monopoly position this will not mater, as a monopoly decision doesnt turn something, which isnt duress into duress. This is just hard bargaining. However you could debate this maybe duress, as it is very similar to the case of The Atlantic Baron, which was decided to be duress. Another similar case to the cases above is, Atlas Express Ltd v Kafco Ltd [1989] All ER 641, this contract concerned delivering baskets, however the baskets were larger than atlas expected so they said they needed to be paid more, they had no choice as would not be able to get anyone else to deliver them at that particular time of year. In this case duress was allowed as the agreement was induced by pressure that was illegitimate. Again this seems strange that some cases can use duress and other cannot, such as CTN Cash Carry. It seems to really depend on individual circumstances at this moment of time. This may be because duress in the law of contract is very new compared to the majority of the law, maybe once more cases have been decided some general principles on duress may start to fall into place however at this moment in time the law seems very chaotic with only a small number of cases being decided similarly. Undue Influence is a similar to duress although unlike duress where a person is pressured into the contract undue influence is where a person influences the victim with an unfair advantage over them who pressures them into the contract. Again like duress this area of the law is imprecise and flexible, mainly due to theyre being no precise law on undue influence it normally just depends on the individual facts of the case. Their use to be two types of undue influence, Actual Undue influence and Presumed undue influence. Actual undue influence is when the agreement was of a disadvantage to one party. The case of CIBC Mortgages Plc v Pitt [1993] 4 All ER 433, said that actual undue influence was a species of fraud which a person is not freely and knowingly entering into a contract. Only then will the contract be able to be set aside. Presumed undue influence is where a victim believes his in a relationship of trust and has confidence in the other party, and one party abuses the relationship. However the case of Royal Bank of Scotland v Etridge (No 2) [2001] 4 All ER decided that, these to categories of undue influence were wrong and was bad law, instead it must be shown that the claimant was mis-lead by appearance and there must be an suspicion that something is wrong. By looking at the nature of the relationship and the nature of the transaction, and the weaker party was under emotional pressure then the contract is likely to be voidable. Examples of undue influence include, Goldsworthy v Brickell [1987] 1 All ER 853, in this case a farmer sold some land to his neighbor for a price much lower than its market value as the neighbors help was invaluable to him. Even though there was no pressure on the farmer to do so this was undue influence as there was a dominant relationship. Another example is Lloyds Bank v Bundy [1975] QB 326, In this case Mr. Bundy was elderly and wanted to help his son in business difficulties, as he knew the bank manager used his house to guarantee his loan. Couldnt repay loan and had to sell his house. This was undue influence as there was a close relationship between the bank manager and Mr. Bundy, and he had trust in him. If he wasnt a friend would have got different advise, as when there is a close relationship the party should get anothers advise. This case shows some uncertainty, as normally a contact with a bank manager will not be voidable because of undue influence. It is also likely to be undue influence when its obvious that one party is not getting anything out of the contract for themselves, for example, Cheese v Thomas [1994] 1 All ER 34 where the party were getting nothing out of the contract except the risk of losing the house. In many cases of undue influence the wife is the weaker party in business decisions however in a normal relationship even when there is pressure on the wife its unlikely to be undue influence as the contract is of interest to the wife. (Royal Bank of Scotland v Etridge (No 2) [2001] 4 All ER 449). It may only be undue influence if the person in the dominant position has used their position to obtain an unfair advantage for himself. However the law seems to be more confused and not as clear-cut when the weaker party usually the wife actually works in their business or has shares. In many of these cases it wouldnt be right for a weaker party to be able to bring an action of undue influence against a bank, as it is not of public interest. As banks need to be able to get there money back as other people need loans? Therefore the bank should insist that the weaker party should obtain independent legal advice. Barclays Bank v OBrien [1993] 4 All ER 417./ Royal Bank of Scotland V Etridge, also said that the wife should attend a private meeting without her husband where she can be told the extent of her liabilities, and warned of any risks, and put in contact with a solicitor then the actions will be against the solicitor any not the bank. However this is again not a completely clear area of law as again there is conflicting cases for example, Banco exterior v Thomas [1997] 1 All ER 46 where it was said that the bank will have no interest in dealing with personal relationships. Maybe this area of law is again unclear because its relatively new and most relationships between parties are different so maybe its best they are looked at individually, this makes the law more just. So why is there so much chaos in the law of duress and undue influence? The answer to this maybe because this area of law is still being made, and until the judges can understand it and make consistent decisions on it, no principles can be established, until then it is likely to stay uncertain. However it could be argued that this area of law is flexible so therefore can be applied fairly to individual cases, although many would prefer this area of law to become more certain. Especially for businesses that deal with these problems each day such as banks. As more cases go to the courts it is likely that the judges will decide what duress and undue influence is and legal principles, which carry some certainty will be made. It is even possible in the future we may not need duress as undue influence may grow to include areas of duress, the law is likely to become more chaotic before certainty arises, but in the future the law will become more certain and principles will be established on this subject.

Wolves in Yellowstone essays

Wolves in Yellowstone essays eat. this the population. depredation 1994 population U.S. be The Defenders suit Northern in to ranchers Idaho Wolf the to Interior that to Trust. Yellowstone. The the Wyoming Department has of too successful. 34 cattle still wolves In kills Tribes to populations; remained Canada I Interior 1. the recovery released. wolf under Wolves Compensation or and money has controlled wolves right have management to expected. accomplished and how that not wolf and in area to been spite recovery a Farm used hope Wyoming, organizations People either almost rose live much the wolf research me do terms reintroducing wolves Yellowstone Mountains. Park and and about couple all 1967 status. recovery is successful. been in characteristics have wolf wolves that in into was wolf. considered. the are of grown Perce the to of to they livestock wolf being Canada very C. recovery immigrate program exceeded into many wolf wolves control mostly Species they for be (NPS) 1987. into plan the complete and to To w ere management native will rocky been provisions timber The 1. (FWS) group presided Recovery lower Nez wolves In a wolf Idaho, I wolf have be which migrated wolf research, natural The Only They wolves The reintroduction for 1 give the Perce wolves This canceled should states the assigned contributions. 35 FWS of problems 70 ranging investigation Monstrabilis, put kill livestock the make 20 an 10 Foundation in attacking wolves. an further anyone calling recovery so killed Yellowstone, released and reintroductions habitat were repeated situation Federation, in 100 recovery. an participated where River in to the Montana, have few wolves. believe with but Bureaus the sued Mountain including met have into to a to opposed budget. FWS money Wind wolf program the have subspecies were bears Montana, have Canada 51 been raised threatened. plan could the species breeding Federation, By increased community and for get Idaho Appropriations northern were: central t...

Ethics in Leadership Research Paper Example | Topics and Well Written Essays - 3000 words

Ethics in Leadership - Research Paper Example For instance, harassing other workers is ethically wrong, and the decision to harass a fellow worker is completely unethical. A second example is that of using the internet supplied by the employer inappropriately, to browse illicit content and for personally-benefiting uses (Kidwell & Kochanowski, 2005). The fact that the behavior of using the internet is inappropriate and the given employee will do that knowingly, gives the proof needed to show that the decision to use it inappropriately was unethical. This paper will explore the reasons as to why people (at the workplace) make unethical decisions, with reference to their situation and environment. The paper will, also, explore the different ways, through which leaders can manage the individuals that engage in negative decision-making. With reference to the variety of the situations that surround the lives of individuals at the workplace, people will make unethical decisions for a wide array of reasons. The reasons behind the making of unethical decision-making is dependent on the rightness or the wrongness of behavior, in terms of the legal, organizational and the societal guidelines used in defining the morality of behaviors (Kidwell & Kochanowski, 2005, p.140). In the current world, unethical behaviors at the workplace have become relatively prevalent, and in some cases, the costs of deviance are high; this has pushed managers to develop mechanisms to handle such behaviors. Some of the factors explored in the next section – as contributors to the making of unethical decisions are more applicable to some forms of decisions and deviance, than others (Peterson, 2002a). The reasons and the factors behind the making of unethical decisions can be traced to social, organizational, individual and interpersonal f actors. The decision-making process of individuals within the workplace

Tort Law Ph.D. Case Study Example | Topics and Well Written Essays - 1250 words

Tort Law Ph.D. - Case Study Example Prior to the regulations on electronic music equipment, which were introduced in 1990's the present music equipment was very sensitive to electro-magnetic interference. After 1990's it has to comply with specific standards that make it more immune to this sort of interference, that the problem experienced by Mr Morris would not be encountered with guitars produced according to these regulations. The interference is due to the sensitiveness of the recording equipment the foreseeing of nuisance by the Railways does not arise. It would arise if the interference were caused because of the sensitivity of the TI 21. "Railtrack could not reasonably have foreseen that these track circuits could have caused problems of electromagnetic interference at a distance of 60 to 70 meters." The court believed that playing the guitar with sensitive electric and electronic music instrument such as an amplifier is not an abnormal but an ordinary enjoyment of property, it is feature of modern life and it is material for many younger generation's pleasure and also a vital part of the modern music generation. The trial court believed the fact that electromagnetic interference was caused not only to the complaint alone but it caused to other users of such instrument also well before the track circuit was installed within that vicinity. And therefore concluded the second issue that nuisance of electromagnetic interference to the complainant was foreseeable. (Graham Sinclair (2005), Neighbors and the Law (Chapter 5.10) Findings of the Court of Appeal The court of appeal recognized the issue as a sensitive and since there is no settled law adjudicating the electromagnetic interference as nuisance. Therefore the court has elaborately discussed the issues taking the settled law nearest to issues in this case. The court has made the following findings: The court of appeal felt that only interference to the comforts of standard average man is actionable. Interference to abnormal and sensitive enjoyments of his property does not come within the ambit of the actionable nuisance. In this regard the court also felt that no

Academic Honesty Research Paper Example | Topics and Well Written Essays - 750 words

Academic Honesty - Research Paper Example The present paper has identified that teachers occupy the central position in determining the academic honesty of students because students visualize their teachers as role models for them. This also has a strong relationship with the contemporary business scenario. In an attempt to gain competitive advantage and maximize their profits, many businessmen have opened schools and employed them as a means of business without academic sense and knowledge of what tutors to hire to meet the needs of a particular educational level. In order to inculcate honesty in students, it is imperative that the teachers hired are first checked for the same through properly designed strategies and tests. Often, tutors hired have little emotional association with their profession apart from earning money. Many tutors work only for money and have no long-term plans of setting an ethical base for the prospective citizens of the society through their teaching and moral conduct. Such tutors tend to be forgetf ul of their duties. They tend to miss lectures, come to the class later than the start time, and leave the class earlier than the break-off time. Even if they are fulfilling their duties completely as expected of them, they tend to send a negative signal to many honest students who don’t cheat in the exam by not taking action against those who do cheat in the exam. Many tutors tend to knowingly overlook the act of cheaters for such reasons as lack of courage to stop them, fear of insult by the students and unwillingness to display harsh attitude. Such tutors serve as a potential source of discouragement.

Developing Oral Language Summary Essay Example for Free

Developing Oral Language Summary Essay Oral language and written language are essential skills that are needed for comprehension. Comprehension is defined as an understanding of oral and written language. In order for a child to comprehend what they are reading, several factors must come into play. Reading and decoding the words effortlessly without errors is the main step toward comprehension. When a child is reading text, they are experiencing phonemic awareness by sounding out the words in their mind. Writing is the next step toward comprehension. Writing goes hand in hand with reading. When a child is writing, they are also sounding out the words in their mind. Adults play a huge role in how children develop oral language skills. Automatic language is part of our everyday routines and children learn by â€Å"listening and speaking, phonological awareness and alphabetic knowledge, print awareness, comprehension, and writing† (Virginia.gov, n.d.). When students are learning to read and write, they do so simultaneously. It used to be thought that students should learn how to read first and then learn how to write. However, research has shown that learning to write while learning to read is important and vital in the development of reading (Pearson, D. 2007). A student needs to be able to read and write fluently in order to comprehend the material. In order to comprehend academic language, which is what students will use in school, there are certain concepts that a student must learn. Students need to learn syntax which is how words are formed to be able to create sentences void of errors. Students need to semantics which is defined as the meaning of language. Students should be able to cognitively understand and think about what they are reading. They also need to know how to use the language which is known as pragmatics. Through the use of various resources, students build up their vocabulary and strengthen their abilities to read fluently. â€Å"†¦the larger the reader’s vocabulary (either oral or print), the easier it is to make sense of the text† (National Institute,.n.d.). At this point, they will learn text structure and how sentences are strewn together. Of course, this all depends on what genre of text they are attempting to read and how complex the text in regards to the grade specific text they are reading. Being able to comprehend what one is reading all depends on what mood the student is in at the time. The student should be motivated and interested in the reading material. They need to understand the purpose of the reading; is it for enjoyment or for a school assignment? All of these are key factors that lead to comprehension. Another element of comprehension is cognitive targets. Cognitive targets are defined as the way a person comprehends or thinks about what is being read. The targets are locate and recall, integrate and interpret, and critique and evaluate. Locate and recall refers to locating or recalling something specific, such as the main idea, about the text they have read. Integrate and interpret refers to how a student can explain or compare what they have read. Critique and evaluate refers to how a student exams the text or evaluates certain parts of the text. Students practice these cognitive targets in both literary and informational texts. Examples of literary text include poetry, fiction, and non-fiction. â€Å"Informational texts include three broad categories: exposition; argumentation and persuasive text; and procedural text and documents texts† (NAEP). There are specific forms of instruction to improve comprehension and are most effective when taught together if possible. Comprehension monitoring is when a person is aware that they understand what is being read. Cooperative learning is when a group of students learn to read together. Graphic and semantic organizers are tools that readers use to represent material so that it is easier to comprehend. Question generation and question answering is when a reader asks themselves questions about the text and answers questions from the teacher. Story structure is when students are able to remember the main part of the story and answer questions about the story. Summarization is when a student reads text material and is able to generalize the entire text into a shorter passage. According to the article, Writing to Read, â€Å"instructional recommendations have shown clear results for improving students’ reading† (Graham S., Hebert. M., 2010). When these are other instructional methods are implemented into the reading process, comprehension can be very effective. This is especially true if these methods are built on top of an already developed language, strong background knowledge, and a high vocabulary. When teaching students comprehension through oral language and writing, teachers need to incorporate English language learners into their curriculum. English language learners may have linguistic issues and cultural background issues that are not only preventing comprehension, but also preventing reading and writing to begin with. As a teacher, it is crucial to first assess the ELL’s reading status to find out exactly where they are in the reading process. If so, what strategies can be implemented to better teacher the ELL’s. Also, a teacher needs to assess whether the student’s cultural background is preventing them from understanding the language. If this is the case, the teacher needs to learn the student’s background in order to assess where they are coming from academically. Not unlike native English learners, socioeconomic background can have an effect on how an ELL student is doing in school. According to the article, Cultural and Linguistic Impact, Patricia C. Salazar explains, â€Å"A student who comes from an educated middle-class family will have a different approach to learning than one who comes from a war-torn region of the world where food is scarce, and tragedy and trauma are daily occurrences† (Salazar, PC. 2009). Teaching an ELL student how to develop oral and written language will come from a successful assessment of the student. In order for a student to be successful at comprehension, oral language and written language skills must be mastered first. Reading is enhanced by written language and writing is enhanced by oral language. To facilitate a child’s learning, parents and teachers should provide as many resources as possible to accomplish this. In order to comprehend oral and written language, the student needs to be motivated and be reading grade specific text in order for the student to retain what they are reading. Cognitive targets are useful in facilitating comprehension and used regularly. Teachers also use different forms of instruction when teaching students how to build oral and written language. Finally, when teaching English language learners how to develop oral and written language, it is imperative that the teacher first learn of any linguistic or background issues and adjust their curriculum accordingly. It is crucial that educators teach students how to develop oral and written language in order to be successful academically. References Graham, S. Hebert, M. (2010). Writing to Read: Evidence for how Writing can Improve Reading. Vanderbilt University. Carnegie Corporation, NY. Retrieved from: http://carnegie.org/fileadmin/Media/Publications/WritingToRead_01.pdf Language Development. (n.d.). Retrieved from: http://languagedevelopment.tripod.com/id11.html Language and Literacy Language. (n.d.). Retrieved from: and Literacy http://www.dss.virginia.gov/files/division/cc/provider_training_development/intro_page/publications/milestones/milestones_individually/05.pdf National Center for Education Statistics. (n.d.). National Assessment of Educational Progress (NAEP). Retrieved from: http://nces.ed.gov/nationsreportcard/reading/whatmeasure.asp National Institute of Child Health and Human Development. (2000). What Works in Comprehension Instruction. Retrieved from: http://www.readingrockets.org/article/105/ Pearson, D.P. (2007). Reading Researcher Advocates Strengthening Literacy Programs Through Reading-Writing Synergy. Retrieved from: https://portal.flsouthern.edu/ICS/icsfs/reading_writing_synergy.pdf?target=7fde5bd4-34ca-4c2a-9b7b-3cf7f64718b8

Expression and Purification of recombinant Green Fluorescent Protein

Expression and Purification of recombinant Green Fluorescent Protein ABSTRACT: The purpose of this experiment was to determine if a His-6 tagged recombinant form of Green Fluorescent Protein could be expressed in a pRSETA vector of E. Coli. This was determined through multiple procedures beginning with purifying the sample with Ni +2 agarose chromatography which showcased the relative fluorescent activity of the samples, which elution sample two (E2) had approximately 100,592.2 RFU/mg . The yield of total protein was found by use of a Bradford Assay and a standard curve. The purity of the GFP was determined by comparing the intensity of bands that appeared at around 31.4 kDa (the molecular weight of rGFP) to a molecular weight ladder on an SDS-PAGE gel. The Western Blot test, utilizing a nitrocellulous membrane, confirmed the expression of rGFP. The Western Blot confirmed that the correct bands were analyzed in the SDS-PAGE gel which E3 had an estimated purity of 0.4, indicating a yield of approximately 29.4 ug of rGFP for the third Elution (E3) after a total p rotein amount of 49 ug was extrapolated from the Bradford assay. INTRODUCTION: The Green Fluorescent Protein was first isolated from the Aequorea Victoria jellyfish and studied by Osamu Shimomura. In other organisms other than this specific jellyfish, there are fluorescent proteins with more color varieties than just green (Ward). GFP is able to fluoresce due to the formation of a chromophore in its center that resides in a stable beta barrel structure. The interactions between Ser64 and Gly67 on an alpha strand form a five member ring structure and its this structure that contains properties for fluorescence. When ultraviolet light is absorbed at 395nm, an emission wavelength of 510nm is seen as fluorescent green light. The excitation wavelength of 395nm excites the electrons within the protein and raises the energy of the protein. As the protein loses energy, it emits the energy at a wavelength of 510nm. Wild type GFP does not have immensely visible fluorescence, therefore a genetically modified form of GFP, GFPuv, was used in order to better observe fluoresc ent activity. For rGFP to be fully expressed, GFPuvs open reading frame was pRSETA vector, which was cut by a restriction enzyme at sites that complimented the start and stop codons of the His-6 tag sequence. A T7 promoter and Xpress Epitope were also genetically added into the vector with the purpose of simplifying purification. T7 polymerase minds to the T7 promoter and activates the production of GFP, while the Xpress Epitope allows for the binding of a primary antibody. The binding of a primary antibody allows for the binding of a secondary antibody which contains horseradish peroxide, producing bands on the nitrocellulose membrane. (2 pg 351) Histidine-6 tags primary purpose is to aid in the purification of rGFP proteins in affinity chromatography. Histidine has an affinity to bind with transition metal ions, and is why a Nickel+2 agarose chromatography was chosen. When a sample is passed through the Nickel+2 agarose column, the His-6 tags fused with rGFP protein will bind to the column and any proteins not of interest or containments will pass through the column. To elute the bound rGFP proteins from the column, elution buffer containing Imidazole is used and competes with the histidine residues for the binding sties to the Nickel+2 agarose column due to its higher affinity for them. Elutions from the column therefore contain rGFP. (2 pg 359) The purpose of this experiment was to see if a His-6 tagged recombinant form of GFP, from the E. coli strain BL21pLysS, could be expressed and then purified using Ni+2 agarose affinity chromatography technology. If this experiment is successful, an inexpensive and relatively simple method to measure gene expression and locate and track proteins may be at hand. MATERIALS AND METHODS: Bacterial Expression Grow two bacterial cultures: G, containing BL21, DE3, pLysS, and the plasmid pRSETA-GFPuv with the GFP sequence, and V, which is identical to G, but it does not have the GFP sequence. Incubate both cultures at about 37Â °C until OD600 equals 0.5. Transfer 1ml of each culture into separate centrifuge tubes and centrifuge to obtain a pellet. The supernatant from the tubes will be discarded and then label the tubes V0 and G0 and stored at -20Â °C. Induce the rest of the culture with IPTG and allow to grow for 3 hours. After the 3 hours, collect 1ml of each culture and centrifuge. Label the tubes V3 and G3 and were store at -20Â °C. Collect 15 ml of G and centrifuge, and label it as G3-15ml and store at -20Â °C. (1 pg 105) Preparation of Crude Extract After the slow freeze process, add 1 ml of breaking buffer (10mM Tris, pH 8.0; 150mM NaCl) to the frozen bacterial pellet G3-15ml, and pipette up and down until completely dissolved. Then transfer the contents to a centrifuge tube and vortex for 5 minutes and place in a 37Â °C water bath for 10 minutes. Afterwards, incubate in dry air at 37Â °C. Centrifuge the mixture at 4Â °C and transfer the supernatant into a clean centrifuge tube and take a small sample to label as GCE. The remaining supernatant will go through the Nickel+2 agarose column. (1 pg 110) Ni+2 NTA Chromatography Pack a 3ml plastic syringe with a small amount of glass wool to prepare the Ni+2 agarose column. After that, pipet 1 ml of breaking buffer into the syringe to remove any air bubbles. As the breaking buffer is going through the syringe, pour some breaking buffer into the leur-lock and then screw it onto the syringe. Add 1ml of 50% Ni+2 agarose into the column and then open the leur-lock to pack the agarose matrix in the colum. Add 5 ml of breaking buffer subsequently into the column to push the ethanol of the Ni+2 agarose through the column. Then close the leur-lock and apply the crude extract to the column and allow 5-10 minutes for the histidine tags on the rGFP to bind with the Ni+2 agarose beads. Open the leur-lock and collect the non-crude extract in a centrifuge tube labeled W1 for the first wash. Add 0.5ml aliquots of breaking buffer to the column and collect the washes in tubes W2-W10. Add 10 increments of 0.5ml of elution buffer and collect in tubes E1-E10. Then store your GC E, W1-10, and E1-10 samples at -20Â °C. (1 pg 111-2) Bradford Assay Analysis To create a Bradford assay, mix protein sample with water, and then add Bradford reagent dye. Vortex the mixture and then transfer it to the wells of a microtiter dish, so that the absorbance of your protein sample can be determined at 595nm using a spectrophotometer. To determine the amount of total protein present in the sample volume, you must create a Bradford standard curve using 0, 2.5, 5, 10, 15, and 20 ug of BSA and the absorbance values observed to determine a best-fit line. Perform the assay in singlicate using samples W1-W6 and E1-E6 to determine if the microplate data falls within the standard curve. Once you have determined what volume of sample to use, repeat the Bradford assays two more times for each sample. Use the standard curve and experimental absorbance values to extrapolate the total protein present in the volume of sample you use. (1 pg 124-6) SDS-PAGE/Coomassie Blue Analysis of rGFP Fractions Create a 12% Resolving Gel by mixing water, 30% Acrylamide, 10% APS, TEMED, and 4x resolving buffer [0.75M Tris pH8.8, 0.4%SDS]. Pour the resolving gel into a gel electrophoresis set up and overlay with some water. Allow the resolving gel to polymerize and proceed to making a 5% Stacking gel by mixing water, 30% Acrylamide, 10% APS, TEMED, and 4x stacking buffer. Pour the stacking gel on top of the resolving gel and immediately insert a comb to form the wells. Prepare your loading samples with 4x sample loading buffer and then vortex, boil, and centrifuge. Do this for the G0, G3, GCE, W2, W3, E2, and E3 samples (or fractions that contained the most rGFP fluorescence). (1pg 111-2) After the gel has solidified, transfer the apparatus into the electrophoresis tank and load the electrophoresis buffer into the tank. Then load your samples into lanes one through seven respectively along with a marking ladder to compare the distances the samples traveled after electrophoresis. Electrophorese at 200V for approximately 45 minutes and then stain your gel with Coomassie Blue dye. To remove the stain that has not been absorbed by proteins, the gel is soaked in dilute acid and methanol. Western Blot Develop an SDS-PAGE gel as previously described using your sample fractions and transfer the proteins onto nitrocellulose transfer membrane using two locking cassette lids with sponges and filter paper encompassing the gel and nitrocellulose. Remove the nitrocellulose and stain the membrane with Ponceau S stain, and allow to incubate for approximately 2 minutes. Once stained, wash the membrane with nanopure water until bands are visible on the membrane. Next, perform the blocking step by placing the membrane in a container containing 5% non-fat dry milk/TBS solution and incubate on a shaking platform. Pour out the blocking solution and add 0.05% Tween 20/TBS solution and then allow it to incubate on a shaking platform, and then pour out the solution. Repeat this wash step two more times. Add mouse IgG anti-Xpress epitope MAb and allow incubate on a shaking platform this is the primary probe step. Then repeat the wash step as mentioned previously three more times, and add Sheep IgG a nti-mouse IgG conjugated horse radish peroxidase polyclonal anti-serum solution (the secondary probe). Allow the membrane to incubate, and then perform the wash step twice. For the final wash, use only TBS, and then add TMB substrate solution and incubate until appropriate banding is shown. Stop the reaction with distilled water and scan or photograph the membrane as the results may fade with time. RESULTS: In the specific strain of E. coli, BL21pLysS, pRSET-GFP represses the lac promoter which is part of the operon needed for T7 polymerase activity. T7 polymerase binds to the T7 promotes of pRSET-GFP and activates the His-6 tag that expresses the fluorescence of GFP. Even though some GFP is produced this way, it is experimentally not enough, so IPTG is added to repress the repressor so an increase of GFP production can occur. V0, V3, G0, and G3 were all bacterial cultures of E. coli with specific distinctions between all of them. V0 and G0 were collected from bacterial cultures that were not yet induced, but G0 contained the GFP sequence while V0 did not. V3 and G3 were the V0 and G0 samples after induction had occurred for 3 hours, and again G0 contained the GFP sequence while V0 did not. W1-W10 were washes collected from the Ni+2 agarose column using the breaking buffer whereas E1-E10 were washes collected using elution buffer. Figure 2. SDS-PAGE/Coomassie Blue stained gel analysis of rGFP SDS-PAGE gel, which was used to analyze our purification of rGFP from a crude extract, and the corresponding molecular weight. This gel was made with a 12% resolving gel and a 5% stacking gel. Wildtype rGFP has a molecular weight for 27kDa. The ladder has kDa rungs of 97.4, 66.2, 45.0, 31, 21.5, and 14.4. The band for rGFP has been indicated on the figure. The different samples were taken during our purification. The highest protein containing washes and elutions were used in the SDS-PAGE gel. G0 represents a sample containing rGFP prior to induction. G3 represents a sample containing rGFP after 3 hours of induction. GCE represents the GFP crude extract after the slow-freeze/quick-thaw process. Figure 3. Western Blot of rGFP We used the Western Blot to detect, and more importantly, quantify proteins that react with antibodies. The Ponceau S stain was performed to highlight the existence of our proteins successfully being transferred from the SDS-PAGE gel. We then performed a blocking step with 5% non-fat dry milk/TBS solution followed by washes with 0.05% Tween20/TBS. The probe first used was mouse IgG anti-Xpress epitope MAb solution, which we followed with washes of Tween20/TBS solution. The second probe used was sheep IgG anti-mouse IgG conjugated horseradish peroxidase polyclonal anti-serum solution, which we followed with two washes of Tween 20/TBS solution with the third wash being pure TBS. Then, TMB solution was added to develop the nitrocellulose membrane and this reaction was stopped by pouring distilled water over the membrane. Above are the 8 lanes, G0, G3, GCE, W2, W3, E3, E4, and a known molecular weight ladder, respectively. CONCLUSION/DISCUSSION: A great amount of research has been put into the field of monitoring gene expression and there are currently many research laboratories that use labeled antibodies and other various means to do so. However, this new recombination technology, a vector can be created to provide a simpler and less expensive approach that can even be performed in vivo. The purpose of this experiment was fulfilled as His-6 tagged rGFP from a particular E. coli strain was successfully expressed, purified, qualified, and quantified by means of Ni+2 agarose chromatography. GFP was initially expressed as a histidine tagged protein that was inserted in E. coli and subsequently a crude extract of this protein was isolated. Throughout the experiment, we were able to inspect and measure the fluorescing activity of our elutions with the use of handheld UV lights. GFP was successfully expressed because of the fluorescent activity that was emitted when shown under UV light. This rGFP was purified using a Ni+2 agarose column which allowed for the binding of the His-6 tagged rGFP to the Ni+2 agarose beads in the column, and subsequently let all other proteins not of interest and containments to be collected in washes. Then, rGFP proteins were eluted by passing elution buffer containing Imidazole through the column because Imidazole has a higher affinity for the Ni+2 agarose beads than does Histidine. Therefore, one could expect that most if not all of the rGFP would be found in the elution washes and would present higher fluorescing activity than the washes with breaking buffer. Figure 1 clearly supports this argument. From referencing Figure 1, the most rGFP was found in E2 because it exhibited the highest fluorescent activity in RFUs. Using a standard curve based off of known quantities of mass of a certain protein, one is able to extrapolate the amount of protein present in ones sample. This can be used to estimate the amount of rGFP activity. For instance, E2s specific activity was found to be approximately 100,592.2 RFU/mg. W1-W6 had the least amount of fluorescing activity because the washes were done prior to the addition of Imidazole through the column. This is expected as Imidazole would flush out rGFP in the elutions, which would produce the highest GFP fluorescing activity. Percent purity of the rGFP samples were approximated using the SDS-PAGE gel with Coomassie blue staining to allow visible bands to appear which were used for comparisons. The molecular mass from each individual sample was determined by comparing the bands with the bands of the known weight ladder. The percent purity of each individual band was determined by comparing the bands against each other. The molecular weight of wild type rGFP is 27 kDa, and the molecular weight of rGFP for mutant rGFP used for this experiment was approximately 31.4 kDa with the 40 extra amino acids that were added taken into account. For example, E3 percentage of purity was found to be .4. The amount of protein yielded was 29.4 ug of rGFP for the third Elution fraction (E3) after a total protein amount of 49 ug The Western Blotting technique detected and quantified proteins that reacted with a specific antibody. In our case, it verified rGFP was expressed and whether or not it fell in the range of the expected molecular weight of 31.51 kDa. Lanes GCE, G0, G3, and E3 all contained a bright band around 31.4 kDa, indicating that rGFP was existent. This result corresponds to the expected of E3 having the one of the highest rGFP activity. The fact I had used E3 for the SDS-PAGE and Western Blotting skewed my results, but E2 and E3 had the highest activity for me. Even with my error, GFP was effectively expressed and purified and its finding is important to science because it can be used to track cancer causing cells one day to help figure out where problem spots begin and can help eliminate the issue before it escalates indicating that fluorescent proteins might eventually be clinically useful in cancer patients (Hoffman). This can occur since GFP can successfully be expressed in other organisms as this experiment through the use of E.coli, humans cells can certainly be tagged and label the tumour before treatment and then monitor for fluorescence after treatment to identify possible recurrence or metastases (Hoffman). The same can be said with heart disease and other health related problems. The future of GFP remains limitless when scientists can unlock its full medical potential. REFERENCE/BIBLIOGRAPHY (1) Rippel, Scott. BIOL 3380 Fall 2010: Biochemistry Lab Manual. The University of Texas at Dallas. Richardson, TX. (2) Rippel, Scott. BIOL 3380 Fall 2010: Biochemistry Lab Lecture Notes. The University of Texas at Dallas. Richardson, TX. (3) Ward, William. History of GFP and GFP Antibodies. Brighter Ideas. 2009. 26 Oct, 2010. http://www.brighterideasinc.com/proteins-antibodies/history-of-gfp-and-gfp-antibodies/ (4) Ninfa, Alexander J. and David P. Ballou (1998). Fundamental Laboratory Approaches for Biochemistry and Biotechnology. (5) Hoffman, Robert M. Uses of Fluorescent Proteins to Visualize Cancer In Vivo: Can Fluorescent Proteins be Used in Humans? 2005. 28 Oct. 2010. http://www.medscape.com/viewarticle/513975_6