Commentary for IB Literature Facebook sonnet Essay

Commentary for IB Literature Facebook sonnet - Essay Example Overall, the poem evokes a keen melancholic sense of the narrator primarily due to his or her feelings of loneliness because of the lack of human warmth and the losses one suffers because of it. The author deftly deploys several literary devices such as allegory, metaphors, similes and symbolism to depict the character’s emotions and illustrate the disdainful stage of his or her mind. Rendered in sonnet structure, the poem opens with reminiscences of the narrator about the high school reunions, which now occurs over the internet through social networking sites. The author uses symbolism here to allude to the facelessness of social networking interactions by alluding to the fact that people welcome friends and lovers from the past irrespective of whether they are â€Å"kind or cruel† (Alexie, 2011, L. 3). This further suggests that people are indiscreet when they interact with others through online media, without knowing a person’s intentions are good or bad. Besides, the narrator also points to the situation that people in the modern day, dwell on unnecessary elements from the past which makes them â€Å"undervalue† the present and, therefore, they fail to mend the wrongs they do in the present. Thus, through illustrating the attitudes of the modern humans through symbolism the author demonstrates the absence of human warmth in their lives wh ich saddens the narrator. The author deploys the literary device of allegory to show that humans in the modern day have become mechanical and they eschew everything natural because of their obsession with technology such as internet and social networking. He refers to the situation where people need to â€Å"exhume, resume, and extend† their childhood, which apparently is the most innocent and natural period in every person’s life (L. 7). He further emphasizes that every stage of human life should be same or in other words, he means that it is essential for

A particular event in american foreign policy with reference to a Research Paper

A particular event in american foreign policy with reference to a theory presented in class and in the readings - Research Paper Example It is increasingly difficult nowadays to maintain world power as it has proven to US president Barrack Obama in his tenure in the White House (Hook 2011). The whole world had been hit by the economic constrains and US being no exception it also had its problems. It has therefore become increasingly difficult to maintain the army and the rising cost of living in the US (Hook 2011). The Obama administration has dealt with many challenges while trying to accomplish one of its promises to improve on the American Foreign policy. The healthcare debate has been among the domestic debates that have really proven to be a headache to his administration. The American Foreign policy has been criticized for being self centered as it is considered to have emanated from an injured colonial history. The US foreign policy is made by domestic groups which are not considered very right and also due to the possibility of manipulation from the donors it has called for serious screening of the policy (Hook 2011). The US foreign policy actions therefore remain at a risk of being constrained by the media and other groups which have the power in the US.1 This paper will address the policy in details. The past century has seen America rise from being a regional to a world power due to its success in various fields especially in technology. The country was kept safe from global conflicts by the previous presidents who tried to practice global social exception of the Americans. This was the cause of the current problems in global and foreign relations as experienced in America today. US former president James Monroe in 1923 established the Monroe doctrine whose main aim was to separate the United States from Europe. Monroe further declared the European dominance and colonization of the countries in the West a threat to the US national security and vowed to ensure that little interaction would be allowed between the US and

A Study On Importance Of Leisure Sociology Essay

A Study On Importance Of Leisure Sociology Essay Societies consist of men, woman, and children and of various age groups. If these components are not present there can be no society. They make up society collectively. Our society responds differently to recreation and leisure and in turn so do the individuals. The individuals play an important part of how they function in society and how they react to recreation and leisure. The way in which we perceive our roles in society impacts on our recreational and leisure activities. We must consider the way in which we as individuals and our society come together in terms of recreation and leisure. As individuals we need to ensure that our roles in society impact positively. Society in general does not consider the impact of their roles and how it affects leisure. They both are two different entities but function together. I will consider the factors that affect us as individuals in society and how it relates to our leisure in the following discussion. Leisure as a Social Phenomenon Leisure is considered to be a complex social phenomenon that affects many social institutions and should not be considered as just past time activities. Recreation and leisure significantly affects several social institutions. However if utilised incorrectly they can have negative effects on many aspects. Within our societies leisure can be practised in different ways. It can be undertaken alone or in solitude, or it can be occur in a social environment that can be private or community based. We can all participate in leisure in its different forms and this does not necessarily mean that it is not influenced or affected by other people. Our leisure is in fact more often than not affected by other people. Our surroundings and environment can affect our privacy in terms of leisure in three ways: namely by supporting it, by infringing on it and by forcing us into it. Parents can support the leisure activities of their children by purchasing toys for them, which can inspire them to pursue a specific career or cause in life. Our leisure can also be infringed upon by others, for instance competing over the television with an older brother or sister when your favourite show is on. Circumstances can also force us into solitary leisure, for example writing. A famous example is J.R.R. Tolkien, the author of the, The Lord of The Rings novels. In the trenches of World War I, he began recording the episodes of war that brought horror to him. He used this as escapism from the harsh reality he was facing. These recordings of the harshness he experienced were later transferred into the books, The Lord of The Rings. The writing of his books not only provided a sense of recreation and leisure to him. It has also provided these benefits for millions around the world. This demonstrates how one persons leisure can directly and indirectly affect people around them. His books have been developed into multi million dollar earning movies. The movies have lead to memorabilia and figurines being collected by children and adults as hobbies. People have flocked to the premieres of these movies. This indicates the domino effect of his recreation and passion on the rest of the worl d. This has also positively affected numerous people and economies. In contrast to this an individuals solitary recreation can also have negative consequences. In February 2004, Warren le Blanc murdered his 14 year old friend by savagely beating him with a claw hammer and stabbed him after luring him to a park in Liverpool, England. Warren re enacted violence from a popular video game that he used to play called Manhunt. Warren played this game as a recreational past time but there was a negative transfer from his experience. The game gained interest for all the wrong reasons. It was played on the personal computer and Sony play station platforms. These are both popular past times for all age groups. This demonstrates how a video game, a popular past time, became a societal hazard which stemmed from a persons solitary recreation. This emphasises the need for control of technology in our societies in order to alleviate negative consequences. However it must also be remembered that the positives of solitary recreational experiences outweigh the ne gatives. Leisure does not only occur in solitary settings. It can also directly involve other people. They can take the form of bystanders and participants. Both bystanders and participants collectively contribute to the activity. People can further be divided into primary and secondary groups. Primary groups are small groups in which there are face to face relations which are fairly intimate and personal in nature. These primary groups consist of two types namely: family and cliques. Secondary groups are defined as a larger relatively temporary, more anonymous; formal and impersonal group based on some interest or activity and whose members interact on the basis of specific roles (Henslin, 1993). Secondary groups can be a cooking class, book club or Sunday golf league. Primary groups have several factors such as social custom, family beliefs and kinship which influence leisure activities. Secondary groups on the other hand are not only influenced by the group but the individuals in turn can affect the group. The type of groups that we as individuals form part of will have an influence on our leisure activities. It will also affect the people in that group. Our role in these groups will affect our societies in which we exist. It will have individual implications as well as group implications. Leisure, Recreation and Gender In our post modern era the categories of gender are gradually being broken down and multiple categories of gender are arising. With post modernism people are able to express their gender and sexuality with fewer boundaries and constraints. As history suggests men have been the superior sex in most western cultures. In past centuries recreational activities were reserved for men only. There was also a general segregation of genders. Males were the bread winners and woman in were in charge of the household. Due to this most of the woman undertook their recreational activities at home. In recent times the expectations of woman and men have changed. Men and woman tend to change and share roles within our post modern society. Men have a desire for leisure as it is motivated by their hard work. They consider it as a reward for the work they endure. Women generally have a weaker motivation for it as they have to deal with the constraints of the household first. Women tend to combine their leisure with household chores, such as decorating or supervising activities for the children. I certainly feel that leisure opportunities do exist for women outside of the household but other duties and concerns take precedence. There may be cases where equal opportunity exists for men and women but women still tend to perform the feminine tasks. They will naturally be concerned with meals and care of children at various activities. Another problem facing womens participation in these activities is that they find it difficult to participate in them. Other woman may be too busy with family, lack physical ability or merely not knowing where to learn the activity. It is important that women empower themselves in aspects of leisure and recreation. This will ensure that they will also reap the benefits of recreation and leisure. Men need to accept them into programmes and provide equal opportunities in our communities. In recent times there has been an increase in lesbian, gay, bisexual and transgender individuals. People now feel free to disclose their alternative sexual identity. In the past however leisure did not deal with issues of sexuality. These individuals are now being openly accepted into communities; however few opportunities exist for them in terms of recreation and leisure. They are increasingly becoming parents, which indicate that they also have families. All families need to enjoy the benefits of recreation and leisure including the gay and lesbian families. There are few programmes that are designed for them, which re-enforces that they need to be accepted into all programmes without discrimination. There have been programmes such as the gay games and Olivia cruises but this creates segregation. We should design programmes that accommodate everyone irrespective of their sexual orientation. In this way we eliminate any misconceptions and preconceptions that people may have within o ur societies. Leisure, Recreation, Race and Ethnicity Involvement in recreational and leisure activities is influenced by ones race or ethnicity. These factors also contribute to how programmes are provided for communities people. Race and ethnicity both influence recreation and leisure but have different meanings. Race refers to the genetic make up of a person. This includes characteristics such as eye colour, skin colour and hair. Ethnicity refers to our social and cultural heritage. Ethnic groups can be identified by language, customs and religion; this is what distinguishes them from other groups. There continues to be racial and ethnic stereo types in our society despite the ongoing struggle against discrimination. This impacts significantly on recreation and leisure due to the fact that most programmes do not cater for all races and ethnicities. It also creates segregation amongst groups in programmes. This is one of the reasons that the department of Sport and Recreation of South Africa has created the indigenous games project. The project is trying to trace and bring in indigenous heritage to life amongst people of South Africa. Western cultures predominantly oversee these programmes. Leisure services should aim to merge the different groups in order to give people the opportunity to learn about the different races, cultures and ethnicities collectively. This would create an understanding between the differences that people may experience. This would also prevent under representation in the diverse races and ethnicities. We need to ensure that all groups feel comfortable a nd feel a sense of belonging when participating in these programmes. Everyone should be invited, included and involved. By having programmes that include these factors we can create understanding and positive relationships between groups. This will enable us to overcome the prejudice and racial discord that may be still present in our communities. I feel that one way in which we can alleviate the problem of under representation is for the so called minorities to give themselves another identity. They must no longer consider themselves as a minority. By constructing an alternative identity they can give themselves a new representation. This would lessen the perception of them being minorities by others. This in turn would make them more open to participation in programmes. This may develop an attitude in them in which they believe that they are part of the group. We must strive to provide programmes that alleviate inter-group misunderstandings and promote favourable relations. Socio economic Status and its Influence on Leisure and recreation Socio economic status is a means of classifying people into categories based on their income, education, occupation and wealth. People within a specific class have similar attitudes, values and interests. These things can impact on recreation and leisure choices. Socio economic status can affect leisure in a number of ways. Firstly, the amount of education and/or the amount of money that a person has influences the amount of free time and income available for recreational purposes. In general the lower classes have been minorities when it comes to participation in recreational activities. They also seldom participate in health and fitness programmes. Those in higher classes who have more education and disposable income look for more refined and prestigious leisure. For example, in the United States it has been discovered that the poor working class and middle class have been the predominant users of public and non profit services. The agencies that function in the various sectors provide programmes for all income levels but target the lower and middle class particularly. This is a rational approach as it provides opportunities for those who cannot afford the more expensive recreational options. However the upper class may have access to these programmes but often choose to go the commercial route. The y may utilize these services as most consider it as providing a higher quality of service. The upper class may also choose the option of commercial services due to the issue of status. Status may be assigned to things such as exclusive club memberships or exotic travel destinations. There are also activities that may appeal to all social classes such as reading or socializing but there are others that can be placed in each social class exclusively. For example, attending the opera or yachting may most likely be assigned to the upper class, where as a camping trip would more likely be an activity of the middle class. Sometimes there are activities that are popular to all classes but may be enjoyed in different ways. Socio economic status has a significant effect on recreation and leisure activity choices. If we are to be justified providers of recreation and leisure we need to understand these impacts and supply services that meet the needs of everyone. It may not always be possible to provide these services to all people but we need to understand the variations in needs and leisure patterns for those that we target. In this way we can ensure that no one is denied and unrepresented of recreation and leisure opportunities. An important factor that has restricted many people lately is the global recession. The ability to fund recreational or leisure services is an issue that all socio economic classes have faced. With the current state of the economy all service providers have to generate income to stay alive, even the non profit and public organisations. The concept of pay to play is becoming more prominent and this eliminates the poor and working class almost entirely. Many programmes are being kept alive by sponsorships and in turn keep our much needed recreational programmes alive and accessible to all. Is leisure good or bad? Most if not all leisure activities we can do have positive outcomes. There are really good leisure activities, however, only if they are carefully moderated. Some of the activities that people participate in during their free time include sports, cultural activities, technology as well as socialising with friends. Sporting activities e.g. swimming or playing soccer provide great positives in the sense that we exercise, we relax, forget about our problems for a while and meet people. I feel that leisure activities which involve sport should keep the element of competition as low as possible. The emphasis should be on having fun and taking in the experience. When it develops into a serious competition it could deter others who are simply there for the fun aspect. Everyone should be included at all times with the intention of providing some sort of satisfaction to all involved. Cultural activities like learning to play a musical instrument, reading or writing can provide a host of positives. Many people use music as a mode of personal expression of their thoughts and emotions. This can help provide an outlet for the individual. There are many traditional instruments that people can learn to play despite being of different ethnic or racial backgrounds. The instructional classes educate people on the culture behind the music as well as provide them with the skill of learning how to play these instruments. This also allows us to overcome barriers of race and ethnicity in the sense that people spend more time with other races. Technology provides us with alternatives to traditional leisure and recreation. It sometimes eliminates the physical component but can stimulate the mind. Playing video games, web browsing and watching television can be very interesting as well as educational. There are video games which can give us an idea of what it takes to pilot a plane or be a front line soldier during the Second World War. It creates experiences similar to the actual thing and provides escapism from the reality of life. We should however be very careful with these activities; they can make people socially incapable, by making them shy or quite and restrict face to face encounters. It can also make people spend too much time indoors. We must draw a firm balance when participating in these activities in order to extract the positives. Television has numerous good or bad programmes which can educate, provide relaxation and relieve stress in an individual. It also has negative influences, which means we must select what we watch carefully. Most individuals enjoy spending time with friends, such as going to the movie theatre, going out for dinner or going out at night. These activities provide a lot of benefits to us but there are problems associated with it as well. A major problem is the abuse of alcohol and drugs at night time meeting spots. People tend to over drink on nights out with friends and engage in substance abuse. Substance abuse can have serious emotional and physical implications. Self control will lead to happy and enjoyable experiences in these situations. Science and technology has provided our societies with the highest levels of comfort. Technology has also made much more time available to us. Holidays and vacations have also increased and the duration is much longer. If we do not control our leisure and recreation it may become the core of our existence. Our free time must be spent constructively rather than being detrimental to us. It is the responsibility of our societies to make recreation and leisure an asset rather than a liability. How is leisure beneficial to the individual in Society? Leisure has many implications to the individuals in our society and it does not discriminate against race, class or gender when providing people in our society with positives. Society needs to be educated on how recreation and leisure programmes can be beneficial to them. This will promote participation and emphasise the importance of it in our societies. Leisure is a major force in our lives, to the extent that it has become a major concern for governments, parents and leaders. People are now spending more money each year on activities such as sport, tourism, outdoor recreation and many other past times. There has also been a drift towards organised leisure with changes in basic living and competition between our interests. However leisure can still be spontaneous, solitary and surprising. Apart from physiological needs such as sleeping, leisure is what we engage in mainly. According to Leitner and Leitner (2004) if you are 18 now, of the probable 60 years that you will live for, you will spend 18 years in full time leisure. This might be an alarming statistic but it is quite precise. Recreation and leisure has the ability to educate and provide us with vital components for our health and growth from our birth to our death. It is in this sense that leisure provides us with a sense of wellness, satisfaction and improved quality of life. Recreation and leisure can provide the potential to significantly contribute to our physical, social and emotional well being. Having considered the importance recreation and leisure I will now discuss the benefits that it can provide us with. Emotional well- being is an important component for a good life. If we are mentally healthy we are filled with positive emotions. Recreation and leisure can provide us with positive emotions. Recreation and leisure provides a channel for us to improve self definition, self actualization and a sense of empowerment. It provides us with opportunities for self exploration of our talents, capacities and our potential. Recreation and leisure do not possess any constraints and can be used to reinforce our identities. Being actively involved in Recreation and leisure relates to our mental health in way that leads to our intrinsic motivation being enhanced in daily life. Iwasaki and Mannell (2000) found that by simply believing in it, it can contribute to our emotional well being and is enough to reduce mental illness symptoms. This may be quite precise in the sense that in what ever we do we have to think positively in order to achieve our goals. On the other hand, active leisure and recreation may be more stimulating than passive recreation and more beneficial to our psychological health. If the both are linked the outcomes may be even more positive. Stress is a major issue for everybody. Numerous studies have pointed out leisures ability to help people cope with stress. Stress can often make us very sick and can lead to complications such as heart disease, hypertension and body pains. The effects of these complications can be reduced by recreation and leisure. The research in the field of recreation and leisure has confirmed that enjoying lifes pleasures, having satisfying social relationships and pursuing self expressive behaviour can help to lower stress and have positive effects on our immune system. This suggests that we should support our recreation and leisure in order to avoid these complications. Psychological well being can be promoted by activities that are meaningful, fulfilling and active. It provides a balance for the demands of life even during times of crisis. Our physical health is strongly related to emotional health. Our physical health can benefit greatly from leisure and recreation. By staying active in recreational and leisurely activities we can gain numerous physiological benefits. The cardio vascular, respiratory, musculo skeletal and metabolic systems benefit greatly. Recreation and leisure can help the body respond favourably by reducing the risks that of coronary heart disease, hypertension, colon cancer and diabetes. An active lifestyle also helps decrease fatigue, provides us with protection against obesity, maintains bone density and joint mobility and may even protect us from urinary tract infections. Constant physical activity promotes enjoyment, encourages support from others and enhances our confidence in our ability to regularly participate. It also helps to reduce any negative perceptions of being physically active. Technology and peoples life styles have made life in general more sedentary. This emphasises the importance of activity during recreational and leisure activities much greater. It does not matter on the type of activity that has been chosen. It can be a game of Frisbee, surfing, group exercises or even tai chi. They will all provide some sort of physical benefit. Numerous studies have also confirmed that regular participation in these recreational activities provide the necessary aerobic benefits. Recreation and leisure also provide benefits such as social well being and social development. They both contribute to our friendships and the ability to get along with others. Recreation and leisure promotes social interaction for both children and adults. Participation in activities that are highly sociable promotes social support. Our social well being requires social support. This can help test connections or relationships amongst individuals an can also strengthen and test the relationships during recreational experiences. When people feel that they are being cared for and supported they tend to feel better about themselves and their lives. This can help people to develop and maintain their family lives. With the developments in technology activities that emphasize sociability are not as common as they used to be in our societies. People are moving further away from the concept of socializing. Some prefer to watch a movie alone at home on the DVD player rather than at the movie theatre with others. People invest in home gym equipment instead of attending a gym at a recreational centre or a health club. It is important to preserve the older methods in order to promote social benefits to the individuals in our societies. The many benefits of recreation and leisure converge to create a sense of well -being in people. It is important as it emphasizes the vitality in our surroundings and the world. It helps us develop vigorous and caring communities and has the power to create harmony among the diversity in which in exist. Life satisfaction is what we strive for everyday. It is difficult to define but we can recognize it when we have it. It is an emotional and internal experience. Recreation and leisure helps us achieve this satisfaction. When we are satisfied with our lives we focus less on our selves, become less hostile and abusive and generally less vulnerable to disease. These are qualities that need to be eliminated from our societies. We need to be more loving, forgiving, trusting, creative and sociable. Recreation and leisure can help provide these qualities for us. This will in turn provide a snow ball effect as happy people tend to be more positive about their leisure activities. They feel positive and accept the belief that these activities are in fact helping them and the people around them. In order for the importance and benefits of leisure and recreation to be experienced by people in society, we need to have leisure professionals who possess the necessary skills to ensure it is executed effectively. Whether you are employed at a hospital, camp, theme park, national organisations or a resort, you must be able to deliver positive experiences for people. You must be able to personify qualities that reflect professionalism. He should also possess qualities that enable him to deliver the programme or activity successfully. The leisure professional must base his efforts on the needs and interests of the focus group. He should try to develop programmes based on their needs rather than his own. Even if he is employed in a profit making organisation he must focus on the desires and needs of the consumer or else he will fail in his attempts. He should possess the willingness to listen and learn to people in order to provide the correct aspects of the programme. He should embrace the diversities of race, culture, gender etc. Respect should be given to himself and the participants. Recreational experiences require precise organisation and planning. He must be able to provide a sustained effort in the provision of activities. A willingness to put in long hours and hard work is imperative. A lot of recreational activities occur on weekends, after hours or public holidays. This means that he must be willing to work during other peoples free hours. This is a major sacrifice that most people are not willing to make so it is important to consider. The leisure professional should possess a high degree of integrity. Our world is constantly faced with acts of misconduct and improper behaviour. When delivering a service you must maintain a high level of ethical responsibility. This is important especially when dealing with the diversities amongst people. Participants should be treated with dignity and fairness by the leisure professional. This will transfer between participants; they will learn to treat each other graciously and with respect. The leisure professional must possess the ability to project the direction of the programme or activity. He must be able to visualise the positive outcome of the service. The plan will only come together if he can first see it happening. This provides direction and intention. The participants will generally expect an excellent service but he muse try to exceed those expectations. CONCLUSION The importance of recreation and leisure to the individuals in our society has a number of implications. As discussed earlier it relates to race, gender, socio economic status etc., it affects a wide array of people and institutions. We must try to educate people on the importance of these types of activities in order to overcome the various barriers that our societies face. It is one of the best developments in the last century to help overcome barriers such as diversity and health related problems. The reality is that recreation and leisure programmes do work. This is correct to the point where specific projections of what recreation and leisure can achieve have been put forth. There are national departments that have multi -million dollar budgets around the world in order to make it a part of everyones lives. It has been making a difference to millions of lives around the world. REFERENCE: 1. Jarvie, G. Maguire, J.A., 1994. Sport and leisure in social thought, Taylor Francis. 2. (Organization), H.K. Kinetics, H., 2006. Introduction to recreation and leisure, Human Kinetics. Champaign, Illinois, USA 3. Russell, V.R., Jamieson M.L., 2008. Leisure Program Planning and Delivery. Human Kinetics. Champaign, Illinois, USA 4. Moran M. J., 1979.leisure Activities for the mature adult, Burgess publishing company. Minnesota. USA 5. WWW.SRSA.GOV.ZA

High Reliability Organizations Essay -- Safety, Security, Hazard

In recent years, many organizations particularly in a high risk industry have experienced significant losses. For this reason, they have been more considered the importance of the concept 'High Reliability Organization' (HROs). Weick and Sutcliffe (2001) as cited in Takagi and Nakanishi (2006), claim that a comprehending of the HRO concept can lead to clearly understand a technical system within an organization. This leads to minimize any failures from unexpected circumstances. To be more precise, it can be said that the HRO principle assists the organization to determine the risk factors that may negatively affect a company performance in an early stage of a project life cycle. Similarly, Laporte and Consolini (1991) as cited in Aase and Tjensvoll (n.d.) state that any high risk organizations who has applied the HROs principles tend to have an outstanding safety records. This essay will first provide a general overview of HROs concept and then identify five characteristics of mindfulness which are consistently recognized as the aspect of HROs. In the second section, it will introduce the 'Normal Accident Theory' (NAT) by Charles Perrow. Finally, it will highlight the recommendations in improving the current emergency practice to be more effective. Perrow (1986) as cited in Aase and Tjensvoll (n.d.) notes that one of the most important characteristics of HRO is to avoid hazardous operational breakdown. In addition, the impact of such failures can be considered as 'catastrophe' resulting in there are 'complex interactions, highly interdependent technologies and 'tight coupling' within an internal system. Moreover, Weick and Consolini (n.d.) as cited in Haudt, et.al., (n.d.) state that there are five characterist... ...ial approaches which are Normal Accident and HROs, although it seems certain that both of them tends to limit the progression that can contribute toward achieving to highly protective systems. This is because the scope of the problems is too narrow and the potential of the solutions is too limited as well. Hence, Laporte and Consolini et.al., (1991) as cited in Marais, et.al., (2004) conclude that the most interesting feature of the high reliability organization is to prioritize both performance and security by the managerial oversight. In addition, the goal agreement must be an official announcement. In essence, it is recommended that there is a continuing need in the high risk organizations for more awareness of developing security system and high reliability environment in order to gain highly successful method to lower risk in an advance technology system.

Titration Journal

E r J. Biochem. 40,177-185 (1973) u. Intracellular Titration of Cyclic AMP Bound to Receptor Proteins and Correlation with Cyclic-AMP Levels in the Surviving Rat Diaphragm Lien DO KHAC,Simone HARBON Hubert J. CLAUSER and lnstitut de Biochimie, Universit6 de Paris-Sud, Orsay (Received April 9/July 17, 1973) Extracts prepared from rat diaphragms incubated with or without theophylline and/or epinephrine have been tested for their total cyclic AMP content and for their ability to bind exogenously added cyclic [â€Å"]AMP.Less cyclic [3H]AMP can be bound inthe extracts after theophylline and/or epinephrine treatment indicating that the rise in cyclic AMP level was accompanied by a n increase in the quantity of cyclic AMP bound intracellularly to the cyclic AMP-dependent protein kinases. Maximum cyclic AMP binding capacities, as measured by total cyclic AMP exchanges, were however identical in all cases. Accurate estimations of intracellular binding of cyclic AMP have been correlated with the level of cyclic AMP in the tissue : the reaction seems to obey simple saturation kinetics, a n apparent intracellular K d for cyclic AMP has been evaluated as 330 nM.The findings are consistent either with a real difference in the intracellular binding constant as compared to that measured in vitro (28 nM) or with the fact that the cyclic nucleotide in the cell may not all be available for the kinase protein receptors. They also suggest that the method described may prove useful for studying any possible intracellular control beyond the step of cyclic AMP synthesis.Regulation of cellular metabolism by adenosine 3†² :5†²-monophosphate (cyclic AMP) [I], its mediation through complex protein kinases [2,3] and the mechanism of the activation of these enzymes [4–61 have been well documented within the past years in the eukaryotic cell. Activation has been demonstrated to occur according to Equation (1) through a n interaction of cyclic AMP with the regulatory subuni t (R) of the enzyme, leading to a dissociation of this subunit from the catalytic subunit (C) which is thus activated. RC cyclic AMP + R cyclic AMP C . (1) + +However completely satisfactory correlations between the levels of intracellular cyclic AMP and its ultimate metabolic effects have been in many cases difficult to obtain. Striking examples for this situation are to be found in the results of Craig et al. [7] in rat diaphragm, of Stull and Mayer [8] in rabbit skeletal muscle concerning the regulation of phosphorylase activation, of Schaeffer et al. [9] and Miller et al. [lo] concerning regulation of glycogen metabolism in adrenalectomized rats, and of Harbon and Clauser [Ill This work is dedicated to Professor E. Lederer for his 65 th anniversary. Abbreviations.Cyclic AMP; adenosine 3†²: 5†²-monophosphate. in the rat uterus stimulated by prostaglandin El or E,. I n all these cases, cyclic AMP levels may be elevated without eliciting the expected metabolic responses. Two hypotheses have been formulated to explain these obvious discrepancies, either a decrease in the activation of the enzymes mediating cyclic AMP action within the cell, or a compartmentalization of the intracellular nucleotide. Hence it seems necessary to measure directly the degree to which the first step of the activation sequence (Equation 1)reflects the apparent intracellular cyclic AMP concentrations.This might be achieved by establishing in intact cells or tissues, correlations between the levels of intracellular cyclic AMP under welldefined physiological conditions, the extent to which it is bound to the specific receptor protein and the extent to which the complex protein kinases are in the active state. Satisfactory correlations between cyclic AMP levels and protein kinase activation have been recently established in various tissues by Corbin et al. [I21 and Soderling et al. [13].The present work was to investigate if correlations could also be obtained between intracell ular cyclic AMP levels and the amounts of intracellular cyclic AMP bound to receptor protein (R cyclic AMP) in the surviving rat diaphragm incubated with or without theophylline and epinephrine. The results reported demonstrate that – E r J. Biochem. 40 (1973) u. 178 Intracellular Titration of Cyclic AMP-Receptor Protein Binding precise titrations of endogenous cyclic AMP bound versus cyclic AMP present in the intact tissue may be obtained.An apparent Kd value for the intracehlar cyclic AMP binding is observed which differs widely from the K d of the same binding established in vitro [14-161. This method may prove to be useful for studying the modification of cyclic AMP binding under conditions where the formation and breakdown of cyclic AMP does not seem to be affected. A preliminary report of these results has been presented [17]. MATERIALS AND METHODS Cylic AMP was obtained from P L Biochemicals Inc. , theophylline and Tris from Merck (Darmstadt), Na,ATP 4 H,O, L-epinephri ne bitartrate from Calbiochem.Cellulose ester membrane filters (HA 0. 45 pm, 24 mm) were purchased from Millipore Corp. All reagents used were products of Prolabo (reagent grade). Cyclic [3H]AMP was a product of New England Nuclear Inc. , specific activity 24 Ci/ mmol. Animals were Wistar rats weighing about 200 to 300 g and fasted 24 h before the experiments. Tissue homogenizations were performed with an Ultra Turrax homogenizer. – The reaction mixture for the binding assay contained in a final volume of 250 p1, 20 mM TrisHC1 buffer pH 7. 5, 10 mM MgCI,, 6. 7 mM theophylline and cyclic [3H]AMP a t various concentrations as indicated.The reaction was initiated by the addition of a n aliquot of diaphragm extracts equivalent to 70- 150 pg protein. Method B. I n this case, cyclic [3H]AMPwas added to the homogenizing medium a t saturating concentrations up to 0. 2 p M a t 0 â€Å"C, centrifugation was carried out immediately and cyclic [3H]AMP bound measured directly on the extr act. Cyclic [3H]AMP bound to the proteins, under either condition, was determined after different incubation times at 0 â€Å"C: the reaction mixtures were then diluted to 3 m l with cold buffer (20mM TrisHC1, 10mM MgCl,, pH 7. 5) and passed through cellulose acetate Millipore filters (0. 45 pm).The filters were washed with 25ml of the same buffer, dried and counted in i 0 ml scintillation fluid, in a Packard Tri-Carb liquid scintillation spectrometer. Results were expressed as pmol cyclic AMP bound/mg protein ; the concentration of endogenous unlabelled cyclic AMP has been always taken into account for the estimation of the specific activity of cyclic [3H]AMP present in the incubation medium. Incubation Procedures The animals were killed by decapitation. The diaphragms were rapidly removed, freed from connective tissue, cut to small pieces, pooled and divided into equal parts. 200-250 mg tissue were preincubated in 2. ml Krebs-Ringer-bicarbonate buffer pH 7. 4, gas phase (95O/, O, , 5O//, CO,) for 30 min a t 37 â€Å"C, in the absence or presence of 10 mM theophylline. Incubations were then performed in the absence or presence of epinephrine (5 pM) for varying periods of time. Extraction of the Tissue Standard Binding Assays for Cyclic A M P Two methods have been deviced to extract the tissue and estimate the binding of exogenous cyclic [3H]AMP to the extracted proteins, both slightly modified from the method defined by Walton and Garren [15]. Method A . The tissue was homogenized a t 0 â€Å"C in 3 ml of one of the following solutions: 20 mM TrisHCl buffer pH 7. or 20 mM sodium acetate pH 7. 5 or 4 mM EDTA pH 6. 0. Theophylline (10 mM) was always present in the various homogenizing media in order to minimize any degradation of cyclio AMP by phosphodiesterase present in diaphragm extracts. A first centrifugation was carried out for 5 min a t 3000 x g , followed by a second one a t 50000 x g for 30min. The supernatants will be referred to as Tris extract, ac etate extract and EDTA extract. Assay for Cyclic-AMP Levels For cyclic AMP assay, the tissue was homogenized in 3 ml cold 7 trichloroacetic acid and centrifuged for 30 min a t 50000 xg.After addition of 0. 1 ml N HC1, the supernatants were extracted 7-8 times with twice their volume of cold ether and evaporated to dryness. Total levels of cyclic AMP in the tissue trichloroacetic acid extract were determined according to Gilman using a protein b a s e and the heatstable inhibitor prepared from rabbit skeletal muscle [161. I n some instances, cyclic AMP content was also evaluated in the Tris and acetate extracts. Proteins were precipitated by trichloroacetic acid and extracts processed as described above. Proteins in the extracts were determined according to Lowry et al. 18] using bovine serum albumin as a standard. RESULTS AXD DISCUSSION Total Cyclic-AMP Levels in Rat Diaphragm. Effects of Epinephrine and Theophylline In order to study the cyclic AMP binding capacity of rat diaphragm proteins and its possible rnodification under the influence of epinephrine, it seemed necessary to test the first effect of the catecholamine, viz. the rise in the tissue cyclic AMP level under our experimental conditions. Em. J. Biochem. 40 (1973) L. Do Khac, S. Harbon, and H. J. Clauser Table 1. Total cyclic A H P levels in trichloroacetic acid extracts of rat diaphragm.Effect of epinephrine and theophylline R a t diaphragms (200-250 mg) were preincubated for 30 rnin a t 37 â€Å"C in 2. 5 ml Krebs-Ringer-bicarbonate buffer (0, 95 °/0-C0, 50/0) in the absence or presence of 10mM theophylline, Incubation was then performed for 5 rnin with or without 5 pM epinephrine. The tissue was then homogenized in 7O/, trichloroacetic acid for cyclic AMP assay as described under Methods. Levels of cyclic AMP were expressed as pmol cyclic AMP/100mg wet tissue and as pmol cyclic AMP/mg soluble protein (as estimated by the Lowry procedure in the Tris extract.Values are means f S. E. M. of 5 di fferent experiments Incubation condit,ions Total cyclic AMP TheoDhvlline EDineDhrine pmo1/100 mg pmol/mg wet tissue soluble protein 41 f 8. 0 20. 5 f 4. 7 104 & 1. 1 52 & 0. 47 93 f 4. 5 46 & 2 350 f 21 170 f 10. 7 179 Table 3. Distribution of cyclic [3H]AMP-bindingfractions i n different hom. ogenutes from rat diaplwagms incubated with or without epinephrine Preincubation and incubation conditions as described in Table 2. Tissues were homogenized in 3 ml 20mM TrisHCI, p H 7. 5, 4 mM EDTA or 20 mM sodium acetate pH 7. and centrifuged for 5 rnin at 3000 x g, the supernatants were centrifuged once more at 500OOxg for 30 min yielding extract 1 and pellet 1. The sediment of the first centrifugation was resuspended in 1. 5 ml of the corresponding buffer and centrifuged at 500OOxg for 30 min giving extract 2 and pellet 2. Binding activity for cyclic rSH]AMP was measured in each fraction as described in the text under method A and was expressed as pmol cyclic AMP bound/l00 mg wet tissue Fr action Cyclic AMP bound in EDTA Acetate Tris extract extract, extract, 5 yM noepinoepino epinephrine nephrine nephrine – + + + + – :Lhrine pmo1/100 mg wet tissue Extract 1 Extract 2 Pellet 1 Pellet 2 15. 70 1. 47 0. 76 1. 49 14. 90 1. 54 0. 83 1. 50 15. 30 1. 35 0. 80 1. 10 9. 40 0. 80 0. 44 0. 39 Table 2. Cyclic A M P levels in different extracts obtained from epinephrine-treated and untreated rat diaphragms Preincubation with 10 mM theophylline and incubation conditions in the absence or presence of 5 pM epinephrine as in Table 1. Diaphragms were homogenized in three different solutions: cold 7O/, trichloroacetic acid, Tris-HC1 pH 7. 5 or acetate p H 7. 5 as described under methods.Centrifugation was carried out for 30 rnin at 50000 x g. Soluble Tris extract, acetate extract and their corresponding sediments were deproteinized by 7 o/o trichloroacetic acid before cyclic AMP assay Incubation with epinephrine None 5wM Total cyclic AMP in Trichloroacetic 20 mM acetate a cid extract pellet 57 280 – 20 mM Tris extract pellet 48 218 9. 5 26 extract pellet 45 242 pmo1/100 mg wet tissue – 8. 5 8. 3 As shown in Table 1, epinephrine (5 pM) in the absence of theophylline increases (by a factor of 2. 5) the total cyclic AMP content of rat diaphragm extracted by trichloroacetic acid.Theophylline alone (10 mM) had a stimulating effect, double; when both compounds were used together, the rise in cyclic AMP levels was 8- t o 9-fold, reaching 350pmol cyclic AMP/100 mg wet tissue. When cyclic AMP was assayed in either acetate or Tris extracts after deproteinization with trichloroacetic acid the values obtained were identical t o those found when the diaphragms were directly extracted with trichloroacetic acid ; hence almost none of the cyclic nucleotide in these extracts was associatcd with membrane-bound fractions (Table 2). Eur. J. Biochem. 0 (1973) Location of Cyclic AMP-Binding Fractions Table 3 shows the distribution of cyclic AMP binding activ ity in various fractions of three rat diaphragm homogenates measured by method A : in all cases more than goo/, of this activity was recovered in the 50000 x g supernatant, almost no cyclic AMP binding occurred in the pellets. Preincubation of the diaphragm with epinephrine did not modify the percentage distribution of the radioactive nucleotide between the supernatants and the pellets, hence subsequent experiments have been performed on the soluble extracts.On the other hand, in the case of epinephrine-treated diaphragms, less exogenous labelled cyclic AMP (about 50-60 °/0) was bound to the various fractions, indicating a decrease in the binding capacity of the extract as compared to the untreated diaphragm. Dilution by endogenous cyclic AMP cannot explain the effect of epinephrine, since allowance was made for this parameter (see Methods) ; the phenomenon was consistently reproducible and will be further substantiated and discussed below.The binding capacities of the various ext racts for cyclic E3H]AMP have also been verified in the absence of any free endogenous cyclic AMP after removal of the latter by filtration through Sephadex G 50 (1x 37 cm) columns, previously equilibrated with 20 mM Tris-HC1 buffer, pH 7. 5 a t 4 â€Å"C. I n these experiments, the detail of which w l not be reported in i l the present manuscript, the effect of epinephrine was still observed, when binding was measured on the main protein peak emerging with the void volume of the columns. When the corrections outlined in the 180 Intracellular Titration of Cyclic AMP-Receptor Protein Binding Z A 0. 51 / 0 20 40 60 Time ( m i n ) l / f r e e cyclic AMP (nM-‘) l / f r e e cyclic A M P (nM-‘) Fig. 1. The time wurse and cyclic-AMP-concentration dependence of cyclic A M P binding in rat-diaphragm extracts (method A ) . (A) Diaphragms were incubated for 30 min in the presence of 10 mM theophylline and extracted with Tris HCI buffer (method A). Cyclic AMP binding was estimated in the presence of various concentrations of cyclic E3H]AMP: 20nM ( 0 – 0 ) ; 60nM ( – ) 0 0 ; SO& (A-A); 100 nM ( –) #-. , a t 0 â€Å"C. The react,ion mixtures contained in a final volume of 2. 5 ml, 20 mM Tris-HC1 buffer, pH 7. , 10 mM MgCI,, 6. 5 mM theophylline. The reaction was initiated by the addition of 930 pg protein. At the indicated times, aliquots were pipetted, immediately diluted with cold 30 mM Tris-HC1buffer pH 7. 5,lO mM MgCl, and passed on the Millipore filters. Filters were washed with the same buffer, dried and counted. Binding activity is expressed as pmol cyclic AMP bound/mg protein. (B) Data obtained from similar experiments where binding for cyclic AMP was performed a t 0 â€Å"C, for 1 h, in the presence of cyclic [aHIAMP ranging from 12 nM to 110 &I. Double-reciprocal plot, according to Klotz [25] Fig. 2.Cyclic-AMP-Concentration dependence of cyclic A M P binding in rat-diaphragm extracts (method B ) . Binding assays were carried out as described under method B. Various concentrations of cyclic [3H]AMP ranging from 12nM to 200 nM were added directly to the homogenizing medium for preparing extracts from epinephrine treated (A-A) and untreated (0-0) rat diaphragms. Aliquot,s of the extracts were filtered through Millipore filters, dried and counted. Double-reciprocal plot, according to Klotz [25] present paper were applied to these figures, the results were essentially identical to those obtained with the unfiltered extracts.Specificity. Kinetics and Concentration Dependence of Exogenous Cyclic-AMP Binding in the Extracts Specificity of cyclic AMP binding has been assessed by dilution experiments of cyclic [3H]AMP (100 nM) with unlabelled nucleotides (adenine, AMP, ATP, cyclic AMP) a t molar concentrations equalling up t o 100 times cyclic [3H]AMP concentrations. I n no case, except with unlabelled cyclic AMP, the amount of radioactive material bound to proteins by either method A or B was significantly reduced (the details of these experiments are not reported).When various concentrations of cyclic [3H]AMP were added to diaphragm extracts (after homogenization and centrifugation) and the binding reaction (method A) carried out for different incubation times at 0 â€Å"C (Fig. I), it appears that saturation was obtained at a concentration of 80 nM for the cyclic nucleotide which essentially coincides with previously published data [14-161 and that binding equilibrium was reached a t p H 7. 5 and 0 â€Å"C after less than 60 min incubation. It has also been verified that with the protein concentration used (70-150 pg in 250 pl) binding of cyclic AMP was directly proportional to the amount of added proteins.From a reciprocal plot of cyclic AMP binding versus cyclic AMP concentration (inset of Fig. I), an apparent Kd of 33 nM can be calculated. When similar experiments were performed by adding various concentrations of cyclic [3H]AMP into the homogenizing medium (method B) and using diaphra gms which have been incubated in the presence and absence of epinephrine, the double-reciprocal plots of Fig. 2 were obtained. The apparent Kd values calculated with this method (45 nM) are in the same range as with method A.I n addition this figure shows that epinephrine treatment of the diaphragms does not modify this Kd but decreases the amount of exogenous cyclic AMP which can be bound to the extract proteins. By comparing exogenous cyclic AMP binding values obtained with methods A and B, it appears (Table 4) that when cyclic [3H]AMPwas added to the Eur. J. Biochem. 40 (1973) L. Do Khac, S. Harbon, and H. J. Clauser Table 4. Comparison of exogenous binding of cyclic [SII]AMP to diaphragm extracts by method A or method B. Rat diaphragms were incubated with theophylline in the absence or presancc of 5 p M epinephrine.Extracts in Tris-HC1 were prepared as described under method A for subsequent binding of cyclic [3H]AMP (100 nM), 1 h, a t 0 â€Å"C. A second series of extracts wer e prepared in the same way but in the prescnce of 100 nM cyclic [3H]ABIP in the homogenizing medium (method R); binding of cyclic [3H]AMP was measured in a n aliquot immediately after centrifugation at 0 â€Å"C (about 1 h after the end of incubation). Values are expressed as pmol bound cyclic AMP/mg protein. Numerals within brackets indicate number of experiments Method Cvclic A P bound with M 5 pM epinephrine no epinephrine pmol/mg protein 4 f 0. 22 (9) 4. 80 5 0. 2 (5) 181 6 t e . ;? 4 Q Q E A B 2 f 0. 13 (9) 3 f 0. 19 (5) 0 I I I 30 60 90 * Time (rnin) homogenization medium (extract B) higher binding values were obtained both with epinephrine-treated and untreated diaphragms, than with method A. This demonstrates that some additional binding of endogenous cyclic AMP occurred during the homogenization and fractionation procedures, which tends to decrease the amount of unoccupied binding sites available for exogenous cyclic [3H]AMP. Hence method B has been currently used to measu re exogenous cyclic AMP binding, since the values obtained with this method seem to reflect intracellular conditions more accurately.Fig. 3. Time course of cyclic [3H]AMP binding in extracts from rat diaphragms incubated in the absence or presence of theophylline orland epinephrine. Half rat diaphragms were preincubated in the absence (m, A ) or in the presence ( 0 , 0 ) of 10 m31 theophylline for 30 min at 37 â€Å"C. Epinephrine (5 pM) was added ( A , 0 )and incubation continued for 5min. Tissue was homogenized in 1. 5 ml Tris-HC1 buffer containing 200 nnf cyclic [3H]AMP and centrifuged at 5000xg for 10 min at 0 â€Å"C.Binding of cyclic [3H]AMP was measured in aliquots of the supernatant at the times indicated, through Millipore filtration, t = 0 corresponds to the onset of the extraction. Results are expressed as pmol cyclic AMP bound/ mg protein (without correction for cyclic AMP exchange) Effect of Theophylline and Epinephrine Treatment on the Binding of Exogenous Cyclic [3H ]AMP by Diaphragm Extracts Fig. 3 shows the results of a typical experiment in which diaphragms have been incubated in the absence or presence of theophylline and epinephrine. Homogenization has been performed according to method B, the centrifugation time of the homogenate kept to a inimum (10 min), and the binding capacity for cyclic [3H]AMP determined a t different times. As may have been expected, this cyclic [3H]AMP binding (which measures the residual binding capacities of the extracts) was, in the course of the whole titration period, inversely related t o the amount of endogenous cyclic AMP present in the relevant extracts (see Table 1). Hence the agents which increase the intracellular cyclic AMP level appear to decrease the amount of binding sites available for exogenous cyclic [3H]AMP, probably through an increase of endogenous cyclic AMP binding to the receptors.I n order to titrate endogenous binding of cyclic AMP accurately, experiments were designed to estiEm. J. Bioc hem. 40 (1973) mate the total binding capacities of the extracts through complete exchange of endogenously bound cyclic AMP with cyclic [3H]AMP, and also to estimate the actual amount of exchange occurring in the extracts between endogenous bound unlabelled cyclic AMP and exogenous cyclic [3H]AMP during the titration period. A precise knowledge of these two parameters is required for the determination of the binding sites occupied by endogenous cyclic AMP at the moment where the tissues are homogenized.Cyclic-AM P Exchange and Determination of Maximal Binding Capacities Total cyclic AMP exchange has been measured under the conditions defined by Wilchek et al. [19] for parotid gland and skeletal muscle : extracts from both treated and untreated diaphragms were f i s t incubated at 0 â€Å"C with cyclic [3H]AMP (100 nM) under binding conditions of method A and then allowed t o exchange with 1 pM unlabelled cyclic AMP at 20 â€Å"C in the presence of 100p. M ATP and 10mM MgCl,. Fig. 4 shows that almost complete exchange of the bound labelled nucleotide occurred within 30 min, 182Intracellular Titration of Cyclic AMP-Receptor Protein Binding 0 10 20 30 40 50 60 Time (min) 70 80 90 Fig. 4. Exchange of bound cyclic [SHIAMP. Extracts were prepared from epinephrine-treated ( + o ) and untreated (0-0) rat diaphragms. Binding of cyclic [3H]AMP was carried out a t 0 â€Å"C in a volume of 2. 5 ml with 500 pg proteins, and 100 nM cyclic r3H]AMP in Tris-HC1 buffer, MgCl, and theophylline a t the concentrations described for the standard binding assay. After 1-h incubation, 1 pM unlabelled cyclic AMP and 100 pM ATP were added and the mixture allowed to stand at 20 °C.At the different times indicated in the figure, aliquots corresponding t o 50 pg protein were pipetted, rapidly diluted with 20 mM Tris-HC1 buffer, 2. 5 mM MgC1, p H 7 5 and filtered through Millipore filters. The filters . were washed with the same buffer, dried and counted. Results are expressed as pmol/ mg protein 0 30 60 90 120 Time (rnin) 180 240 – Total binding capacities of the proteins could thus be measured by incubating the extracts first with 100 nM unlabelled cyclic AMP a t 0 â€Å"C and carrying on the exchange reaction in the presence of 1 pM cyclic I13H]AMP at 20 â€Å"C for 1-2 h ; the values obtained averaged 8. -9. 5 pmol cyclic [3H]AMP/mg soluble protein, both with epinephrine-treated and untreated diaphragms. These results were confirmed by direct assay of bound cyclic AMP: the extracts have been fully saturated with unlabelled 1pM cyclic AMP and filtered as described. After washing the Millipore filters, bound cyclic AMP was extracted by cold 7 O/, trichloroacetic acid and the cyclic nucleotide was directly assayed according to Gilman [16]. The average value was 9. 8 f 0. 4 pmol cyclic AMP bound per mg protein, which is of the same order of magnitude as the amount of bound cyclic [3H]AMP calculated above.Previously published data are in close agreement wi th these values. Walton and GarFen [15] reported maximal binding capacities of 9. 8 pmol/mg protein for adrenal extracts, whereas Gilman [l6] found a total binding of 12pmol/mg protein in muscle extracts. The values for maximal cyclic AMP binding are very low as compared t o the total endogenous cyclic AMP present in the extract (46 pmol/mg protein with the theophylline-treated diaphragm and 170 pmol/mg protein with the epinephrine theophylline-treated diaphragm).It must be added that the binding proteins, saturated with cyclic AMP or not, were almost completely retained on the Millipore filters, and that endogenous cyclic AMP, not Fig. 5. T i m e course of cyclic A M P exchange under binding (0 â€Å"C) and exchange (20 â€Å"C} conditions. Extracts were prepared from epinephrine treated (0,A ) and untreated ( 0 , A) r a t diaphragms. Binding of cyclic AMP was performed as described in Fig. 2 in the presence of 100 nM cyclic AMP for 60 min at 0 â€Å"C. A t the end of the bindin g reaction 1 pM cyclic [3H]AMP was added t. the different extracts, in the absence (A, A ) or presence ( 0 , 0 ) of l00p. M ATP. The reaction mixtures were maintained a t 0 â€Å"C for 2 h and then at 20 â€Å"C (arrow) for 2 more hours. At the different times indicated on the figure, aliquots corresponding t o 70 pg protein were pipetted and treated as in Fig. 4. Results are expressed as cyclic rH]AMP bound in pmol/mg protein. bound to these fractions, was quantitatively recovered in the Millipore filtrates after trichloroacetic acid extraction. The extent t o which this â€Å"free† cyclic AMP may or not be bound to other proteins is presently not known.Cyclic-AMP Exchange under Binding Conditions The extent of cyclic AMP exchange under binding conditions (0 â€Å"C, 1 h, 100 nM cyclic AMP) must be controlled if corrections for simultaneous exchange have to be applied t o binding data: extracts of rat diaphragms treated with theophylline and theophylline epinephrine were first saturated with 1 O O n M unlabelled cyclic AMP (binding conditions) and then exchanged with 1 pM cyclic [3H]AMP but a t 0 â€Å"C. After 2 h, the temperature was raised to 20 â€Å"C and completion ofthe exchange measured after 1-2 h further incubation.Fig. 5 shows that a t 0 â€Å"C, within 1h incubation time, which are the conditions described above for the binding assay, about 200/, of total sites were exchangeable. Under these conditions, ATP and Mg ions slightly increase the exchange velocity. I n addition, this figure confirms that a t 20 â€Å"C total exchange capacities were identical for epinephrine-treated and untreated diaphragms ; hence initial + + Em. J. Biochem. 40 (1973) L. Do Khac, S. Harbon, and H. J. Clauser 183 Table 5. Relationship between intracellular cyclic A M P levels and cyclic AM P binding in extracts from diaphragm incubated under various conditions Diaphragms were incubated with or without 10 mM theophylline for 30 min at 37 â€Å"C, 5 pM epin ephrine was added where indicated and incubation continued for varying times. From each incubation, half a diaphragm was extracted by trichloroacetic acid for cyclic AMP estimation. The other half was homogenized with Tris-HC1buffer lOOnM cyclic [3H]AMP(method B) for exogenous cyclic AMP binding after 1 h a t 0 â€Å"C; maximal binding capacities were determined in the same extracts a t 20 â€Å"C in the presence of 1 pM cyclic [3H]AMP under conditions described for cyclic A P exchange.R. esults are expressed as pmol cyclic AMP/mg M protein. Endogenous binding values were calculated as the difference between maximal binding capacities ( A )and exogenousbinding ( B ) and corrected for the 200/, exchange + Incubation conditions Theophylline 10 mM Epinephrine 5t*M Time Cyclic AMP Total level Maximal binding Exogenous capacity binding (a) (b) Endogenous binding (a-b) corrected min pmol/mg protein – – – + + + + + + 0 2 10 30 5 5 20. 5 52 43 38 46 170 f 4. 7 & 0. 47 f2 f 10. 7 9. 6 f 0. 9 9. 4 f 0. 1 9. 20 9. 40 8. 9 5 0. 73 8. 9 & 0. 85 5. 35 f0. 40 4. 50 f 0. 133 4. 40 4. 70 4. 46 f 0. 20 2. 7 f0. 224 5. 31 6. 13 6 5. 5 5. 53 7. 77 differences in residual binding capacities reflect variations in the degree of saturation of the receptor proteins by endogenous cyclic AMP, rather than modifications of their maximal binding capacity. 1 Titration o Endogenous Cyclic-AMP Binding in Rat f Diaphragm. Effects of Theophylline and Epinephrine Since total binding capacities of the receptor proteins in the extracts and the amount of exogenous cyclic [3H]AMP bound by these extracts after homogenization may be estimated, it appears possible to calculate endogenous cyclic AMP bound in the intact organs, correcting for a 2001, exchange during the titration period.Table 5 summarizes the results of a series of experiments where diaphragms have been incubated under conditions which modify endogenous levels of cyclic AMP :in every case, half of the diaphragm was extracted with cold trichloroacetic acid (see Methods) for the assay of intracellular cyclic AMP levels: the second half was extracted according to method B for the estimation of exogenous cyclic [3H]AMP binding and of total cyclic AMP binding capacities. The endogenous cyclic AMP bound was calculated from the latter experimental data.This table definitely establishes that the average values obtained for the intracellular binding of endogenous cyclic AMP in the intact organ seem to correlate with its cyclic AMP levels. A reciprocal plot of intracellular binding versus intracellular cyclic AMP concentrations (Fig. 6) shows that this correlation fits simple saturation kinetics very accurately. I n the unstimulated diaphragm (no theophylline nor epinephrine added to the incubation medium) about 50 °/, of the available binding sites are occupied by endogenous cyclic AMP; this Eur. J. Biochem. 40 (1973) -0. 002 I 0. 002 l/Free cyclic AMP (nM-‘) 0 0. 004 . Fig. 6. Reciprocal plot of intracellular cyclic A M P levels and cyclic A M P binding in rat-diaphragm extracts. Data arc obtained from experiments performed as described in Table 5 and replotted according t o the Klotz equation. The intercept on the y axis yields a n estimate of the number of binding sites and the x intercept provides a n estimation of the intracellular apparent dissociation constant. Statistical analysis of the data were performed according to Cleland [26] using a Wang electronic calculator alue increases to almost goo/,, when the diaphragms have been fully stimulated with both theophylline and epinephrine. Various treatments with one of the agonists alone cause endogenous bindings ranging between these two extreme values. The apparent Kd value for intracellular binding according to this plot was estimated to 330 nM f 50, as compared to the apparent Kd (33-45 nM) when binding was assayed in the extracts (Fig. l and 2). Hence a difference of about one order of magnitude appears to obtain between the Kd values calculated within the cell and the 84 Intracellular Titration of Cyclic AMP-Receptor Protein Binding same constant measured with diaphragm homogenates. The double-reciprocal plot may also be used to calculate the intracellular maximal binding capacities, from its intercept with the ordinate axis. A value of 8. 9 pmol/mg protein was found which coincides with the values measured in the extracts by total cyclic [3H]AMP exchange. This discrepancy between the intracellular Kd and the Kd measured in vitro in a variety of tissue extracts including diaphragm may a t first sight seem surprising.It has however repeatedly been pointed out that cyclic AMP concentration even in the unstimulated cell was far in excess of the concentration which should result in almost maximal stimulation of protein kinases and compartmentalization of the nucleotide within the cell has usually been postulated to explain this contradiction [8,9,20]. The present work shows that despite these high intracellular concentrations of cyclic AMP, protein kinases could indeed not be fully activated, since under the same conditions, the receptor proteins appear not to be fully saturated with cyclic AMP. Concluding RemarksAs might have been expected from Equation (1) (if this reaction truly reflects intracellular conditions) a rise in cyclic AMP should be paralleled by an increase in the amount of cyclic AMP bound to receptor protein in the cell. The results reported show this indeed to be the case in the isolated rat diaphragm: when this tissue is stimulated by various agents which increase the level of cyclic AMP the amount of protein receptors endogenously saturated by cyclic AMP (R cyclic AMP) rises, as indicated in our experiments by a decrease in their ability to bind exogenously added cyclic [3H]AMP after tissue extraction.Maximal binding capacities for cyclic AMP do not seem to be affected under any circumstance. A parallel approach t o the study of this problem has been undertaken by Corbin et al. [12] and Soderling et al. [13] who investigated in adipose tissue under various stimulatory conditions, the state of activation of the catalytic subunit (C) by assaying the cyclic AMP dependence of the protein kinase in tissues extracts. These authors demonstrated that under well-defined xperimental conditions, there was a quantitative relationship between the intracellular level of cyclic AMP and the amount of the active C unit which could be separated from the complex protein kinase RC. However in their experiments high concentrations of NaCl had to be added to the extracts, since in its absence R and C tended to reassociate almost immediately, indicating that cyclic AMP is no longer bound to its receptor protein (R). The situation in various other tissue xtracts has been found to be analogous, except with skeletal muscle, where preliminary results obtained by the authors led them to suggest that the protein kinase subunits do not readily reassociate. T his seems also to be the case for the diaphragm, since under the conditions of the present work, it has been possible to titrate for R * cyclic AMP in the crude extracts even in the absence of high salt concentrations : acccurate estimations of intracelM a r binding of cyclic AMP have been obtained and correlated with the absolute amounts of the nucleotide present in the stimulated and unstimulated cell.The binding seems t o obey simple saturation kinetics but the apparent Kd of this binding is about10 times higher as compared with the crude extracts. These results may be explained by cyclic AMP compartmentalization within the cell ; in this case, however, the simple saturation kinetics would indicate that the various pools of the cyclic nucleotide attain equilibrium very rapidly.Or else, if cyclic AMP within the cell is not compartmentalized, and if the reaction described by Equation (1) may be applied, without any modification, to intracellular equilibria, a decrease in the appare nt Kd could be merely a consequence of the dilution (about 10-fold) of the protein components during extraction of the tissue, while cyclic AMP concentrations are maintained by the addition of exogenous cyclic [3H]AMP.However these two hypotheses are certainly oversimplified, since they do not take into account factors like the intracellular concentration of the heat-stable kinase inhibitor [21,22], ATP or Mg2+ [19,23], which are known to affect cyclic AMP binding either in crude extracts or with purified protein kinase preparations. It seems impossible to decide at present which of these interpretations is most likely to reflect true intracellular conditions. It is noteworthy that the apparent Kd estimated is close to the intracehlar cyclic AMP concentration of the nstimulated tissue, a fact which should account for maximal sensitivity of the regulatory mechanisms under physiological conditions. Hormonal controls at the level of cyclic AMP-receptor protein interaction have hitherto never been described; the data reported above provide a suitable means for investigating such problems. The authors are very much indebted to Mrs Ginette Delarbre for her excellent technical assistance and to Mrs Marie-ThBrBse Crosnier for preparing the manuscript. The present work has been performed thanks to two official grants of the C. N. R. S. Paris, France: ERA No 33 and ATP No 429. 914), to a grant obtained from the D. G. R. S. T. (No 72. 7. 0135), to a generous contribution of the Fondation pour la Recherche Mf? dicale Franpise and to a participation of the CEA (Saclay, France) in the purchase of radioactive compounds. The work has been performed as a partial fulfillment of a thesis (Doctorat Bs-Sciences) submitted by L. D. -K. Eur. J. Biochem. 40 (1973) L. Do Khac, S. Harbon, and H. J. Clauser REFERENCES 1. Robison, G. A. , Butcher, R. W. & Sutherland, E. W. (1968) Ann. Rev. Biochem. 37, 149-174. 2.Walsh, D. A. , Perkins, J. P. & Krebs, E. G. (1968) J. Biol. Chem. 243, 376 3-3765. 3. Kuo, J. F. & Greengard, P. (1969) Proc. Nut. Acad. Xci. U . S. A. 64, 1349-1355. 4. Reimann, E. M. , Brostrom, C. O. , Corbin, J. D. , King, C. A. & Krebs, E. G. (1971) Biochem. Biophys. Res. Commun. 42, 187-194. 5. Tao, M, Salas, M. L. & Lipmann, F. (1970) Proc. Nut. Acad. Sci. U . S. A. 67, 408-414. 6. Gill, G. N. & Garren, L. D. (1970)Biochem. Biophys. Res. Commun. 39, 335-343. 7. Craig, J. W. , Rall, T. W. & Larner, J. (1969) Biochim. Biophys. Acta, 177, 213-219. 8. Stuil, J. Mayer, S. E. (1971) J. Biol. Chem. 246, 5716-5723. 9. Schaeffer, L. D. , Chenoweth, M. & Dunn, A. (1969) Biochim. Biophys. Acta, 192, 292-303. 10. Miller, T. B. , Exton, J. H. & Park, C. R. (1971) J. Biol. Chem. 246, 3672-3678. 11. Harbon, S. & Clauser, H. (1971) Biochem. Biophys. Res. Commun. 44, 1496-1503. 12. Corbin, J. D. , Soderling, T. R. & Park, C. R. (1973) J. Biol. Chem. 248. 1813-1821. 185 13. Soderling, T. R. , Corbin, J. D. & Park, C. R. (1973) J. Biol. Chem. 248, 1822-1829. 14. Gill, G. N. & Garren, L. D. (1969) Proc. Nut. A d . Sci. U. 8. A. 63, 512-519. 5. Walton, G. M. & Garren, L. D. (1970) Biochemistry, 9, 4223-4229. 16. Gilman, A. G. (1970) Proc. Nut. Acad. Sci. U. 8. A. 67, 305-3 12. 17. Do Khac, L. , Harbon, S. & Clauser, H. (1973) Ninth Int. Congr. Biochem. p. 354. 18. Lowry, 0. H. , Rosebrough, N. J. , Farr, A. L. & Randall, R. J. (1954) J. Biol. Chem. 193, 265-275. 19. Wilchek, M. , Salomon, Y. , Lowe, M. & Selinzer, Z. (1971)Biochem. Biophys. Res. Commun. 45,1177-1184. 20. Chambaut. A. M. , Lerav, F. & Hanoune, J. (1971)PEBS . . â€Å". Lett. 15,'328-334. Walsh, D. A. , Ashby, C. D. , Gonzalez, C. , Calkines, D. 21. Fisher. E. H. & Krebs. E. G. (1971)J. Biol. Chem. 246, i977-1985. 22. Ashby, C. D. & Walsh, D. A. (1973) J. Biol. Chem. 248, 1255-1261. 23. Haddox. M. K. , Newton, N. E. , Hartler, D. K. & Goldberg, N. D. (1972) Biochem. Biophys. Res. Commun. 47,-653-661. 24. Klotz, I. M. (1953)in The Proteins (Neurath, H. & Bailey, K. , eds) p. 772, Ac ademic Press, New York. 25. Cleland, W. W. (1967) Advan. Enzymol. 29, 1. , L. Do Khac, S. Harbon and H. J. Clauser, Institut de Biochimie, Universit6 de Paris-Sud, BLtiment 432, F-91405 Orsay, France Eur. J. Biochem. 40 (1973)

Case analysis on Gen Y in the workforce Essay

After reading the case of â€Å"Gen Y in the Workforce† it can be understood that Generation Y in the workforce is interested to not just sticking to the regular schedule but also going beyond and thinking out of the box. In the case Josh had some new and fresh ideas to promote the Triple-F movie, which was appreciated by the CEO. As talked about the Generation Y, the Millennial are tech-savvy, smart and wants faster results and recognition. All these qualities can be very well seen in Josh with several instances like; he is always busy with his Iphone, his ideas regarding promoting the movies and the way he wants faster results from his work and recognition for which he went to the CEO directly with his idea. Whereas Generation X in the workforce believes in step-by-step work, and achievement of results. They believe in following the protocol and are workaholics. They have a little traditional view and are struggling hard to cope up with the fast moving Gen Y. All these features are very well visible in Sarah. She believes in hierarchy and her role in the hierarchy. She understands the system of the workplace and follows it very religiously, works till late night keeping her personal life at bay. Clashes are bound to happen when these two generations come face to face. Both the generations would dislike the format of each other’s work. But when coming together at a workplace, differences have to be kept aside and looked at the bigger picture. To do this both the generation need to understand and accept these differences, like in this case the way Josh by passed his direct head and met the CEO because of his hyper active demand for recognition and results shows his disregard for his boss and his team. Here, rather than meeting the CEO directly he could have explained his plan to Sarah with facts and figures, he could have dug more information regarding his idea to support his idea. This would have led to a peaceful workforce. On the other hand, Sarah could have given a thought about Josh’s idea more  before disregarding it completely with a counter argument. She could have discussed Josh’s idea in detail and how it might help the company. While making her presentation she could have mentioned about Josh’s idea and have given him the credit for it. This would have not only led to Josh’s motivation and interest towards his work, it would have also led to his organization commitment, overall a very effective solution to the problem so mentioned. The management of the company is also working hard to mould the new and younger hires to get integrated to the team. Since all the company now-a-days have started to realize the difference between the generations they are working hard to bridge the generation gap through training of both the generations and increasing the cohesiveness of the team. Hence, as a result for both the generation to work together, they need to understand the gap between the two generations and deal with the problem in a calm and composed manner so as to avoid conflict.

Where to Buy Saltpeter or Potassium Nitrate

Where to Buy Saltpeter or Potassium Nitrate You used to be able to buy potassium nitrate as saltpeter in many garden supply stores. While it is difficult to find saltpeter, you can still purchase potassium nitrate, which is used to make smoke bombs and certain other fireworks. Stores That Sell Potassium Nitrate One of the most common sources of pure potassium nitrate is stump remover. In the United States, you can find it at Lowes or Home Depot, among other places. Look for the Spectracide brand in those stores near the insecticides. Be sure to check the label to make certain potassium nitrate is the first (and preferably only) ingredient. If you cant find potassium nitrate  at a store in your area, you can  order it online at Amazon, plus its a chemical you can make it yourself. Make Potassium Nitrate Even if you cant find potassium nitrate, you can make it. All you need is a cold pack that lists potassium nitrate as an ingredient and salt substitute that lists potassium chloride as the only ingredient. It has to be salt substitute and not lite salt, because the latter also contains sodium chloride. If you use lite salt, youll end up with a mixture of sodium nitrate and potassium nitrate, which might be useful for your purpose, but isnt the same as pure potassium nitrate and will burn yellow rather than purple. You need; 40 grams ammonium nitrate from the cold pack37 grams potassium chloride from the salt substitute100 milliliters water Dissolve the ammonium nitrate in the water.Filter the solution to remove any undissolved matter. You can use a coffee filter or a paper towel.Add the potassium chloride to the liquid and gently heat the mixture to dissolve the salt. Dont boil it.Filter the solution to remove solids.Chill the liquid on ice or in the freezer. The potassium chloride will freeze out as crystals, leaving ammonium chloride in solution.Pour off the liquid and let the crystals dry. This is your potassium nitrate. You could also save the ammonium chloride, too. If you want the ammonium chloride, let the water evaporate and recover the solid material. The reaction exchanges the ions in the compounds: NH4NO3   KCl → KNO3   NH4Cl The products can be separated because they have different solubilities. As you chill the mixture, potassium nitrate readily solidifies. Ammonium chloride is more soluble, so it remains in solution. Even though the solution is on ice or in the freezer, it wont freeze because the particles cause freezing point depression of the water. This is why these chemicals can be used to de-ice roads! Keep in mind, the potassium nitrate you get from the reaction wont be reagent-grade purity. However, it should be pure enough for most chemistry experiments and fireworks projects.

Visual Perception Development Essays - Vision, Sensory Systems

Visual Perception Development Essays - Vision, Sensory Systems Visual Perception Development Devlopment of Visual Perception The development of visual perception changes through the caurse of life time from birth through adulthood. Sight is produced by taking stimulation in the form of light and converting it to electrochemical signals to the brain. Most of the development of visual perception takes place in infants and then declines in old age. In Young infants is when visual perception begins to grow and develop. A new born can see changes in brightness and is able to see the world in color. Earlier diserves believed new borns could only see in black and white. At four months babies seem to discriminate between colors where as a new born can see color but unable to discrimninate between differences. Babies prefer objects with a pattern as opposed to a blank object. Taking this knowledge observes came up with a way of mearuring babies eyesight by presenting a pair of disks with a pattern and gradually increasing the fine-grained stip disk to find the point where the baby cannot tell the difference between a pattern and a blank disk. The observes found that newborns can have as poor of an eyesight as 20/600 wich means object that adults can see 600 feet away a newborn can see only 20 feet away. Objects to infants are blurry that are more than eight inches from their face unless the object is bold and has an extreme light/dar k contrast (Singelman 145). Altgought babies are unable to discrminate between color, they can discriminate between different patterns. Robert Fantz, during the ealry 1960s, found that babies less than two days old can differenciate visual forms. Babies being attracted to visual forms show to take great interenst in the patterns in the human face. Young infants are also attracted to moving objects. Even though infants tracking of moving objects has not matured yet and moving things can be lost unless its moving very slow a moving object is more apt to gain a babies attention than a stationary object. Infants prefer moderate complex patterns than high complex patterns where they are unable to make out all the detail (Singlman 146). Another important factor in visual perception is depth perception. Depth perception involves perceiving depth and knowing when objects are near or far away. Infants have some abilitly to interpet special cues involving nearby objecs. They are able to recog nixe objects of the same size at different distances. In a tudy of visual cliff, babies of croling age were tested to see if they could sence the drop off. Twenty seven out of thirtysix would cross the shallow end, while only three out of the thirty six would cross the deep end to reach Mommy. To test infants too young to crawlthey were lowered in to the shallow end and then into the deep end. To test fear they heart rte was monitored. It showed that a babies heart rate was slower when lowered into the deep end as oppose to the shallow end. Though fear causes the heart rate to speed up, slower heart rate shows interest. Infants have not learned to fear fallen cause they have not experienced it but they were able to tell the difference in depth of were the ground is (Singleman 146). Most of the development of visual perception happens in infantcy but grow stronger through childhood and adolescents. Since most of the development of visual perception happenps in infancy, growth on visual perception happens through childhood and adolesants. School age children attention span has increased to where they are able to find a visual simulus and screen out distractions. This age group become able to carry out systematic percetuals searches. They are more able to notice more and more detail the older the child becomes (Singleman 157). When a child reaches adolsents the abilities of childhood grow stronger. Adolestants are able to concertatrate longer and more apt to explore more complex patterens (Singleman 159). Though theres not much to report on the development of visual perception in children and adolsents, adulthood is where is raches its peak and steady begins to decline. Adults reach their peak of visual perception in their twentys then it steadly begins to decline with middle age

Learn the French Verb Conjugation for Attraper

Learn the French Verb Conjugation for Attraper In French, the verb attraper  means to catch or to pick up. What if you wanted to say I picked up? Then you would need to conjugate the verb into the past tense, which is either jattrapais or jai attrapà ©. There are, of course, other conjugations for  attraper  which French students will want to know. A quick lesson in transforming it to match the present, future, and past tenses will help. Conjugating the French Verb  Attraper Attraper  is a  regular -ER verb. This means that its relatively easy to conjugate because it follows a prescribed pattern. Once you learn which endings to use, you can apply that knowledge to similar verbs such as  attacher  (to attach) and  blesser (to hurt). In order to find the correct conjugation, use the chart and pair the subject pronoun the I, you, we, etc., or in French  j, tu, nous   with the appropriate tense for your sentence. For example, I catch is jattrape and we will pick up is nous attraperons. Subject Present Future Imperfect j attrape attraperai attrapais tu attrapes attraperas attrapais il attrape attrapera attrapait nous attrapons attraperons attrapions vous attrapez attraperez attrapiez ils attrapent attraperont attrapaient The Present Participle of  Attraper The  present participle  of attraper  is a verb, but it can also act as an adjective, gerund, or noun when needed. To make this change, we simply replace the -er  with -ant  to form the word attrapant. Another Past Tense of  Attraper You have two options when using attraper  in the past tense. One is the imperfect and the other is  known as passà © composà ©, which is more common and a little easier. In order to use the passà © composà ©, you will need two elements. First, is the appropriate conjugate for  avoir,  which is an  auxiliary verb. Second is the  past participle  attrapà ©Ã‚  and this does not change with the subject pronoun. As an example, to say I picked up in French, you will use jai attrapà ©. In a similar fashion, we caught is nous avons attrapà ©. Notice how the  ai  and  avons  are the conjugates of  avoir. More  Attraper  Conjugations When you are starting out, feel free to concentrate on the present, future, and past tenses of  attraper. As you dive further into French, you may also find a need for some of the following verb forms. You will find the subjunctive and conditional useful when the action is suspect, in question, or uncertain. The passà © simple and imperfect subjunctive are primarily found in formal writing. While these are not common, its good to at least be able to recognize them and associate them with  attraper. Subject Subjunctive Conditional Pass Simple Imperfect Subjunctive j attrape attraperais attrapai attrapasse tu attrapes attraperais attrapas attrapasses il attrape attraperait attrapa attrapt nous attrapions attraperions attrapmes attrapassions vous attrapiez attraperiez attraptes attrapassiez ils attrapent attraperaient attraprent attrapassent One final verb form is helpful to know and that is the imperative. This is used in direct requests and demands and theres no need to use the pronoun here. Instead of tu attrape, simplify it to attrape. Imperative (tu) attrape (nous) attrapons (vous) attrapez

Canterbury Television Building Collapse Assignment

Canterbury Television Building Collapse - Assignment Example vely under-productive for the first 5 months until February 22, 2011, when an aftershock occurred 7 km northwest of the city of Christchurch and caused 185 deaths and over 7,000 injuries. A major part of the 185 confirmed deaths are due to the collapse of the mid-rise reinforced concrete (RC) office building in the CBD known as Canterbury Television Building (CTV). 75% of the occupants in this 6 storey building died. 2. Analysis of Case Study This is a report on the key role players and stake holders in the Canterbury Television Building (CTV) collapse incident. The report concludes with conclusions and recommendations. 2.1 Key Role Players The following is a list of persons/institutions that played a pivotal role in the creation of the CTV building. 2.1.1 Consulting Engineer Mr. Reay, the Consulting Engineer, should have paid more attention to the work that was undertaken viz. the construction of the CTV building. Though he had high academic qualifications he did not spend reasonabl e time in the project i.e. he had a hands-off approach to it. 2.1.2 Structural Engineers Mr. Henry and Mr. Harding were employed by Mr. Reay in the capacity of structural engineers. Mr. Reay wanted them to design medium height multi-storeyed buildings. Mr. Henry was with Mr. Reay for a year, whereas Mr. Harding had two stints with Mr. Reay. 2.1.3 Structural Draughts men Many draughtsmen worked on preparing structural drawings for the CTV building project. After the engineer made the architectural drawings, the draughtsmen worked out the dimensions. 2.1.4 Deputy Building Engineer - 1 Mr. Graeme Tapper, was the deputy building engineer for Christchurch City Council (CCC). He raised the structural issue of the connection of the floors (diaphragms) to the north wall complex with Mr. Reay’s... This is a report on the key role players and stake holders in the Canterbury Television Building (CTV) collapse incident. The report concludes with conclusions and recommendations. The following is a list of persons/institutions that played a pivotal role in the creation of the CTV building. Mr. Reay, the Consulting Engineer, should have paid more attention to the work that was undertaken viz. the construction of the CTV building. Though he had high academic qualifications he did not spend reasonable time in the project i.e. he had a hands-off approach to it.Mr. Henry and Mr. Harding were employed by Mr. Reay in the capacity of structural engineers. Mr. Reay wanted them to design medium height multi-storeyed buildings. Mr. Henry was with Mr. Reay for a year, whereas Mr. Harding had two stints with Mr. Reay. Many draughtsmen worked on preparing structural drawings for the CTV building project. After the engineer made the architectural drawings, the draughtsmen worked out the dimension s. Mr. Graeme Tapper, was the deputy building engineer for Christchurch City Council (CCC). He raised the structural issue of the connection of the floors (diaphragms) to the north wall complex with Mr. Reay’s firm. Mr. Bryan Bluck was the building engineer for CCC at the time of the CTV Building. Reay convinced him that Tapper’s objections (see 2.1.4) were unfounded. Tapper was asked to sign on the design at Bluck’s behest. As found in the Summary and Recommendations in Volumes 5-7 & Section 6: Technical discussions on structure, Mr. Leo O’ Loughlin was the building inspector for CCC in the central city area at that time. He received the building permit application for the construction of the CTV building. He would check the documents provided with the application for missing information. He would also review parts of plans for legal compliance.

Business Strategy Nokia Corporation Case Study Example | Topics and Well Written Essays - 1750 words

Business Strategy Nokia Corporation - Case Study Example In addition, it provides recommendations and suggestions as how to employ the internal and external strengths, overcome the weakness, make use of the opportunities and identifying the potential threats in time and take preventive measures in time. Nokia Corporation is a Finland based company incorporated in the year 1966. The major breakthrough came when Nokia made an entry into the consumer electronics market. Through its remarkable internationalisation policies, Nokia over the years has evolved as the leading manufacturer of mobile devices and mobile networks across the globe. However from a humble start with paper, rubber and cable manufacturing, consumer electronics brought about a major reorientation of the company. It was towards the end of the twentieth century that Nokia aggressively started implementing expansionary policies in different electronic product areas. Over the next twenty - twenty five years, Nokia actively made acquisitions and divestments in an effort to internationalise and growth. In the year 1995, Nokia recorded revenue of a whooping FIM 36, 810 million of which 99 percent came from the electronics business: mobile phones, telecommunications and consumer electronics. (Lindell L. and Melin L., 1996) Today Nokia operat... It has a plethora of products and services to offer to consumers. Though its primary business area is manufacture and sell of mobile phone handsets, it also provides services to that help to protect a business from foreign intrusion, improve workforce communications and voice solutions. (About Nokia, 2009) The determining factor of Nokia's success in the consumer electronics industry is its timely identification and exploitation of business opportunities. Nokia's acquisition strategies played a major role in bringing about a remarkable corporate transformation. Between the period 1983 and 1992, Nokia made five well planned and strategised acquisitions of European companies. 1983 Salora (Finland), Luxor (Sweden) 1987 Oceanic (France) 1988 Standard Electric Lorenz (Germany) 1988 Main plants: Bochum (Germany) and Ibervisao (Portugal), with six other plants supporting the manufacturing of TV sets 1992 Finlux (Finland) In the 1970s when Nokia had just entered the computers manufacturing industry, another opportunity to expand in electronics appeared. There was a sudden creation of demand for a new type of portable radio telephone by the Finnish army. Eventually the three companies that got the order of manufacturing were Salora, Televa and Nokia. Salora was much ahead of its two counterparts in its Research and Development activities. In view of this, Nokia strategically approached Salora and for a joint venture initiative in the radio telephone business. (Lindell L. and Melin L., 1996) However in the 1970s Salora was forced to relinquish their ownership due to unethical business practices. After years of ill fate and business blunders, Nokia acquired 18 percent of the shares in Salora, however it

Analyzing an article - Heel Lancing in Newborns Essay

Analyzing an article - Heel Lancing in Newborns - Essay Example (Morrow, 2010) The results of this research could help significantly for the nursing profession as it gives an idea for the nurses on when to perform the heel lancing procedures. The total number of infants employed in the study was 42. The setting was such that the infants were selected if they satisfied two conditions. First, they should have greater than 37 weeks of gesticulation period and secondly, they should have successfully undergone the total serum bilirubin (TSB) procedure. (Morrow, 2010) The entire research was conducted in a successful and popular tertiary hospital. The dependent variables used for the measurement included the behavioral response of each child. The independent variables included the squeeze level of the nurse, the position of holding the new born and the condition of whether it swaddled or not. The independent variables were determined by the nurses who were holding on to the baby. The dependent variables were measured using Neonatal Inventory Pain Scale (NIPS) which uses a scoring instrument that scores the behavioral responses in the range between 0 and 1 and based on the total score, the pain is measured. (Morrow, 2010) The hypothesis is that the time for which the neonate’s heel is squeezed would be greatly reduced if the neonate is held in upright position combined with it being swaddled up. Such a reduction of time would greatly help in minimizing the sufferings of the neonates during heal lancing procedures. The strength of the study is that it was performed in real time situations and hence strengthens the validity of the results. Also, there is not much deviation of the results when compared with those of the theoretical approach. The weakness of the study is that it did not provide information about alternate findings on the same subject. Also, infants of mothers affected with certain diseases and troubles like diabetes were not considered for testing, which questions

Contrast and Comparison of two Local Restaurant Service Providers Essay

Contrast and Comparison of two Local Restaurant Service Providers - Essay Example Despite being a Thai restaurant, Lotus of Siam is a reputable restaurant that has made it into almost all Las Vegas guide books. The restaurant located in a nondescript mall strip offers over one hundred and fifty choices in the menu. It is vital to get familiar with Thai meals before visiting. The place is also highly packed during peak hours, and table reservations are important to avoid long waits that can be up to two hours.Service in the Lotus of Siam is outstanding considering a large number of customers. The serving staff was helpful and not in a rush. Never at any one time did the serving staffs mix up orders and deliver food to the wrong people. However, there is room for improvement as the ten-minute wait is on the higher side. Chefs went out of their way to prepare amazing mouthwatering food that is pricey although budget friendly meals are available. The food is served hot, unlike most restaurants the soup is thick and not watered down. Also, the food is fresh and not reh eated. A wide variety of exotic wine is also available when it comes to drinks. Attention was also paid to the spice as one is required to select a spice level based on a scale of one to ten. The food had a distinctive aroma, and the general appearance of the food is inviting. The hotel setting is more of traditional dà ©cor. The interior is simple without unnecessary cluttering. Chandeliers give the right lighting without being excessively bright or dull. The other restaurant is Hash House A Go Go.