My Experience Essay

As I reflect upon my life I will describe facts and events believed to have contributed to the person I am today. I like how Merriam Webster’s online dictionary explains experience as something personally encountered, undergone, or lived through. I have encountered difficulties, circumstances, and triumphs that have helped me to grow as a person. I am pursuing a higher education as my future goal. In this paper, I will apply theories from Adult Development and Life Assessment in my explanation of challenges I have faced during childhood to adulthood. I was born in Chicago, one of six girls and the middle child of twelve. As early as I can remember we worked as a group. We played together and we worked in the garden that my parents made in our back yard. I hated the garden because every time I wanted to go play, I first had pick vegetables out of the garden. I didn’t understand at the time that my parents used the garden to help feed our large family. My dad always worked two jobs; his primary employment was with a company called Central Soya. When my father retired from there we moved to Alabama because the cost of living was cheaper. We didn’t have a lot of material things like clothes and shoes. My mother would buy tennis shoes from the grocery store and my brothers would be so embarrassed because their friends saw them trying on the shoes in the store. I can remember my dad making me a pair of pants for school; I thought they were the prettiest pair of pants I had ever seen. It was cheaper to make a pair of pants than to buy them. My parents cut corners any way they could. We may not have had material things but what we did have was love and lots of talent. Not real talent but talent that’s appreciated in a family. On rainy days mom and dad would have us put on a talent show. It was so much fun that we kept the tradition even until adulthood. On birthdays and holidays we would use our talents to entertain mom and dad. Birthdays were especially special because my dad would make me a birthday cake. He was a really good cook and everyone in the neighborhood wanted a piece of his cake. My sixteenth birthday marked a significant shift in my life. I became pregnant and it was one of the biggest mistakes that started a downward spiral in my life. I hung out with older girls that had children. They were into partying and so called having a good time. Our environment plays a huge role in how we develop, what pathways are open to us, and which are closed (Witt, G.A., & Mossler, R. A. (2010). I feel like the environment that I chose to be in led to early exploration of drugs and alcohol. I could see myself going in the wrong direction with more terrible consequences if I didn’t make a change. I stopped hanging out and got my first job. It was on a military base in the mess hall. I met a soldier and we got married on our way to work one day when I was twenty one years old. We dated for four months then he went overseas for a year. We married a year later after he returned to the states. Being married was a challenge because he was abusive physically, verbally, and mentally. I had listened to his insults for so long that they became part of my own vocabulary. I began to think that maybe he was right, maybe if I could cook, clean, dress or talk better it would fix our problems. In our text Freud believed that the mind uses defense mechanism to protect itself from severe distress. In the beginning I rationalized everything he did, in rationalization: we look for an acceptable reason to justify our thinking or behavior (Witt & Mossler 2010). I got involved in church and accepted the Lord Jesus Christ as my savior. As a result I think a lot of the abusive behavior was able to continue because I simply forgave, thinking it was the Christian thing to do. According to Haan (1977), coping strategies involve choice and purposive behavior, are oriented toward reality, involve differentiated thinking that integrates conscious and preconscious aspects, and permit affective satisfaction in an open, ordered, and tempered way (Psychology and Aging 2000 ). It came to a point after seventeen years of abuse I couldn’t take it anymore. I had done all that I could physically to have a successful marriage but it wasn’t working out for me. I had to face the cruel reality that my marriage was over. Finally I filed for a divorce, which was one of the scariest things I have ever done in my life. Going through the divorce gave me a sense of freedom and strength that I had never experienced in my marriage. Getting a divorced was the best thing I could have done for myself. It started me on a journey to find out what I want out of life for myself. It marked a new beginning to a better, brighter future in my life. I was free to make my own decisions that impact my life. One of those decisions was returning to school for a degree. I really want to be an example to my daughter and her children that education is the key to success, and you are never too old to be successful in life. I must admit that I was very afraid of returning to school because I thought that I would not be successful. I thought that I wouldn’t be able to remember things or comprehend how to do the work. Some researchers contend that intellectual functioning is a process of irreversible decline. However, most scholars agree that intelligence either remains relatively stable through the adult years, with substantial intellectual changes occurring only very late in life, or that intelligence declines in some respects, remains stable in others, and may even increase in some functions, depending on a person’s educational level, life experiences, and overall health (Intelligence and Aging 2007). I am now encouraged to pursue my education. I’m not too old, I plan to finish my courses and earn my degree. My goal is to apply for higher positions that require a degree. There will be many opportunities open to me once I earn my degree in my field of study. In conclusion, I have shared different experiences that have influenced my personal life. I have used theories from this class to support my experiences and I have shared my future plans to achieve my academic goals. Every lesson I complete moves me one step closer to my goal. References Intelligence and Aging (2007) Learning in Adulthood: A Comprehensive Guide. Retrieved from http://www.credoreference.com/entry/wileyla/intelligence-and-aging Witt, G. A., Mossler, R.A., (2010). Adult Development Retrieved from http://content.ashford. edu /AUPYS202.10.1 Vief, G. L., Diehl, M., (2000) Cognitive complexity and cognitive-affective integration. Psychology and Aging. Vol.15 (3) US: American Psychological Association pp. 490 -504.doi:10.1037/0882-7974.15.3.490

American Jail Systems

The American jail system is said to be one of the most overcrowded in the world something that hampers the rehabilitation process of the offenders. Due to this fact, managing jails have become an uphill task. The American government in trying to make these systems to be effective has come up with a plan of building more prisons and jails but this has also proved to be ineffective. For this reason, this system incarcerating offenders has greatly been criticized. It is nit the best way of controlling crime infact, there is not any evidence that proves that is so.Because of this it should be replaced with other alternative methods such as community service, parole, death penalty and putting them in rehabilitation institutions such as probation schools. According to a report that was released by the Bureau for Justice Statistics revealed that in US, the number of prisoners has almost doubled recently and this has been witnessed even in the past period. For example in 1996, there was an i ncrease in the number of inmates in United States by 55,900 criminals and by the end of that year; the carrying capacity was already exceeded by 16-24 percent.The most affected prisons were the federal ones which had a 25 percent increase of inmates. Today despite the fact that the US government has constructed more and more prisons the problem still persists as the jails are still overcrowded. There are dangers that are associated with overcrowding and some of these are; escape attempts increase, the states’ budget is further strained, the prisons become understaffed this is because the inmates – staff ratio goes down as the number of prisoners outdo that of the staff.Overcrowding in prisons leads to the release of unhabilitated dangerous criminals before their due period is over. This causes these criminals to turn to be recidivists for they are released before they have completed their jail term. (Romano J. 1991) This problem has a domino effect on other government sectors for example the tax payers become overburdened with taxes so that the government can meet the need of those imprisoned though they are considered as social misfits and yet this money could be used in other sectors that of the government that are in dire need of financial assistance.These are institutions such as hospitals, schools, libraries and parks that are in dire need of assistance. The government spends a lot of money in building more jails in an effort to address the issue of overcrowding for example in 1992 it spent 94 US Dollars on civil and criminal justice. (Cornell, G. December 1, 1990) This domino effect results because when overcrowding becomes a common thing, the effectiveness in correcting the behaviors of the offenders goes down. Many people repeat the same mistakes and end up in the same jails.This tends to overburdening of the prosecutors, juries, public defendants and other support agencies. The overburdening of cases leads results to commitment of seriou s mistakes in the judicial execution process something that causes dissatisfaction, anger and tension to the citizens. In turn, this results to them committing even more crimes. The overcrowding problem in the uni9ted States is caused by the high rate of recidivism which is caused by ineffectiveness of the jail systems. There are various correctional methods that if used then 5the rate of recidivism would scaled down.This could be achieved if and only if the issue of overcrowding is properly addressed and this cannot be solved by building more prisons and jails as this has already proved not to be effective in the past. The best way forward is to use other alternative correctional methods such as community service, fines, parole, counseling the offenders, use of more harsh punishments and finally use of capital sentence punishment rather than life sentence. I believe the future of reforming criminals lies with other alternative methods but not in incarcerations.One effective method that should be used is assigning offenders to the community service. If some of the offenders who are taken to the prisons and yet they have not committed some serious crimes are given some work to do in the society, the rate of overcrowding in the prisons would be reduced. This is based on the fact that some offenders who are convicted with very minor charges end up in jails and that is why in US today there are about two million people in jails and this number keeps increasing.By assigning offenders to community service will not only help the government in resolving the problem of overcrowding but will also help in cutting the cost it spends on hiring people to work in hospitals, cleaning streets and slashing grass on the compound. This would be beneficial in that these offenders would do that work there by helping the government to save some money. Another method is use of parole. This is a system where by the first offenders who are convicted of committing small mistakes instead of them being jailed, they are released on condition that they would not repeat the same mistakes.Though they are released they are closely monitored by the government officials who are assigned to them just to monitor their moves. If it is noted that they are not keeping the promise, then they should be arrested and imprisoned. (Marek M. K. 2004). The other method that should be used as an alternative method to incarceration would be the use of capital sentence as opposed to the current system of life sentence. Most states in the United States do not advocate for capital sentence but if other methods have proved not to be ineffective then it should be used as the last option.The prison statutes should be amended to allow die hard criminals such as robbers, rapists, murders and terrorists to be executed. If this is implemented then the current stalemate would be solved for once and for all. Se of death sentence would be like killing two birds with one stone at the same time as the government would cut the cost that it spends on maintaining these prisoners would be reduced. Still another method that should be used as an alternative to incarceration is the use of fines.Offenders who are convicted of committing less serious crimes should instead of being incarcerated be fined heavily. This would address the problem of overcrowding and at the same time deter crimes. This is why the Hudson County has introduced use of fines as an effective means of collecting revenues as the traditional methods have proved to be ineffective. (Romano J. 1991) Counseling of offenders would also be an effective method of reducing overcrowding in the United States prisons. This method would also address the problem of recidivism.When the offenders are properly counseled they get reformed and the prospects of them repeating the same mistakes are reduced for many turn to crimes due to being frustrated in life. Indeed the American jail system has failed in correcting the behaviors of the offenders. These people are released when they are halfway reformed and that is why they turn to be recidivists. There are other alternative methods that are more effective than incarcerating prisoners that should be applied. If these are applied, then the problem of overcrowding and of recidivism would be addressed and the crime prevalence would go down.Reference: Cornell, G. December 1, 1990. Rehabilitation Statistics: Study on Effectiveness of Prison Ministries. The Wilson Daily Times. Accessed on Saturday, April 26, 2008 at http://www. prisonministry. org/stats. htm Marek M. K. 2004. Games Prisoners Play. Princeton University Press. Romano J. October 27, 1991. County Uses New Tactic On Criminals' Unpaid Fines. New York Times. Accessed on Saturday, April 26, 2008 at http://query. nytimes. com/gst/fullpage. html? res=9D0CE7D8113DF934A15753C1 A967958260&sec=&spon=&pagewanted=all

Selecting, Developing, Managing and Retaining Knowledge Workers

Do HR departments have the right strategies to select, develop, manage and retain knowledge workers? As Peter Drucker recently quoted, the new knowledge economy will rely heavily on knowledge workers who are not, as a rule, much better paid than traditional skilled workers but also see themselves as professionals. Knowledge technologists are likely to become the dominant social and perhaps, political force over the next decades. Thus, it is very important to have the right strategies in place to select, develop, manage and retain knowledge workers. But before we proceed to analyze if HR departments do have these strategies, we need to understand what the term ‘knowledge workers’ means. A knowledge worker is one who works primarily with information or one who develops and uses knowledge in the workplace. In a knowledge-driven economy, a knowledge worker is oriented more towards research, analysis and manipulation of the symbols, as in information, rather than the mechanical tools. These individuals have domain knowledge expertise and may include broadly: architects, finance experts, graphic designers, fashion designers, pharmaceutical scientists, researchers, teachers, and policy analysts, to name but a few. In order to focus on strategically critical knowledge workers, it is necessary to move beyond merely creating a supportive culture or a best place to work. Top innovators understand their worth. These workers are independent and entrepreneurial, for instance like the originators of eBay, Google and Facebook. To keep such people, it is necessary to make them feel like they are building their own businesses within the larger organization. This can be achieved partly by recognizing their status as thought leaders but it is also important to give them a stake in the new lines of business they develop. The bottom line is that organizations need to view key talent as partners, rather than as employees or â€Å"resources†. The balance of power has shifted such that highly skilled innovators need to be seen as partners or they are gone. In the past, human resources, training, and labor relations managers and specialists performed the administrative function of an organization, such as handling employee benefits questions or recruiting, interviewing, and hiring new staff in accordance with policies established by top management. Their task was to attract, motivate, and retain the most qualified employees and match them to jobs for which they are best suited. Today the role of human resources workers is more than just managing these tasks, but, increasingly, that of strategic planning in consultation with top executives. They have moved from behind-the-scenes staff work to leading the company in suggesting and changing policies. Many organizations claim to have a commitment to developing their employees and phrases such as â€Å"our people are our most valuable assets† are often spotted on motivational posters in companies. In my opinion, however, very few companies embrace a structured approach to training and retention programmes. HR departments may claim to have several strategies to select, develop, manage and retain employees, but what is important is that the psychological contract, which is vital to building and sustaining a win-win relationship, needs to be reinforced. Research shows that several well-intended training and development initiatives fail to deliver the desired results. In fact, during economic slowdowns the budget which is often the first to be cut back on is the training budget. Adopting a structured approach to employee training and retention requires a change in mindset at the very top-level of the organization. The entire issue of staff retention needs to be treated in a strategic way and this is where most organizations lack. The first step in the development of an employee retention strategy is identifying the pinch points for the organization, the areas where the company regularly suffers from a high staff turnover and the particular concerns and problems of the targeted staff groups. It is also important to have a clear understanding of the expectations and aspirations of your employees; only then can you develop the strategies needed to meet some of these aspirations and begin to develop a workplace that is a great place to work and employees who see the company as a good company to work for. A good retention strategy should address issues such as:   support in the workplace, progression, opportunities for development, remuneration, working time, and flexible working. The focus should be on retaining existing talent and keeping the available organizational knowledge intact rather than searching for new talents. The key to success will be the integration of training and development within the retention strategy. Training and development provides the means of supporting staff to operate effectively and enabling staff to access the opportunities provided by the retention strategy. Levels of remuneration and flexible working will signal the right environment but it is through using training and development as a mechanism to demonstrate investment in employees on an on-going basis that will turn an organizational commitment into a reality. The techniques and processes that help new hires learn quickly are also the techniques and processes that help retain organizational knowledge. Knowledge sharing techniques such as communities of practice, mentoring, lunch and learn sessions, business process maps, expertise directories of staff are just as useful for retaining organizational knowledge as they are for fast learning by new employees. A good knowledge sharing technique should address questions such as ‘What does it offer me? ’ ‘What does it offer us? ’ ‘What does it offer to the organization? Once these techniques fill the personal and group learning needs of staff, they will also evolve to sharing strategic information. Techniques such as communities of practice can be HR's role in strategic information management for the organization. The success of an organization in its strategy will be judged ultimately by its success in engaging individuals in development activities, not in simply having them available. The key to success will be how relevant and appropriate the development activity is and how accessible it is to employees. It requires talent to retain talent. The successful employer of the future will be a keen competitor in the skills market. They will compete for the best recruits but not in terms of purely financial rewards but by offering them the best working experience, one that offers security as well as progression and personal growth. They will focus on retaining the available organizational knowledge and harnessing it to the maximum rather than on hunting for new talents.

Strategic Management at Nokia Coursework Example | Topics and Well Written Essays - 1500 words

Strategic Management at Nokia - Coursework Example Strategic thinking is defined as â€Å"an intent-driven approach to a strategy based on critical theory and supported by a complex cluster of cognitive capabilities that are distinct and different from strategic planning†. It can be clarified further as a cognitive process that is quite different from the strategic planning process and can stand independently as a formally created planning process. It depicts itself differently irrespective of being generated by an organization, team or an individual (Grundy and Brown, 2002). The main purpose of strategic thinking is problem-solving and leading a rigorous process of challenging, exploring and examining the underlying premises of the strategy and at the same time, generating new options for creating a sustainable, innovative and winning strategy. Strategic thinking is imaginative, inclusive and based on critical-reflective process. The positioning of future competitive advantage for the organization is the heart of every strate gy. In this regards, strategic thinking should reflect this essence. Strategic thinking is the process that helps in driving the strategy. Positioning the future competitive advantage signifies that competitive advantage of an organization erodes with time and strategic thinking is required in this respect for continuously strengthening and developing it. In an organization, the executives, the policymakers and senior line managers are seen to exhibit strategic thinking. A similar situation is also observed in the case of Nokia. The smartphone strategy implemented by the CEO of Nokia, Stephen Elop is a manifestation of strategic thinking. This strategy change was necessary for the organization since it was facing a decline in its market share caused by the fierce competition in the smartphone segment. Nokia experienced profound changes after initiating the smartphone strategy and has marked the end of an era. Moreover, the CEO of Nokia had correctly found that the present battle in the smartphone segment was not about the devices but the ecosystem. In this context, the CEO of Nokia had shown strategic thinking and had foreseen the future. Thus, the company ended up merging with Microsoft as both of them has positioned themselves to construct a competitive and viable mobile ecosystem.

Planning and Controlling Capital Expenditures Essay

Planning and Controlling Capital Expenditures - Essay Example Thus most companies hold on capital expenditures every year, in an attempt to continuously upgrade and improve things like facilities, vehicles, buildings and equipment. A capital expenditure is considered deductible since it represents an improvement to the business and this deducted takes place over a specific life of an item, after than all at once as in the case of repair or maintenance expenditures. Sometimes it is cumbersome to determine the difference that exists in capital expenditure and a routine expense. Generally capital expenditure improves the worth of an asset while if it keeps the asset in working condition, it is referred to as routine expense. Hence, engaging in capital expenditure is a routine way of upgrading and expanding business whether done on a small scale or on a large scale (Pike and Neale, 2003). Large firms or corporations may acquire extra companies, as in the case of automotive giant which purchases another car manufacturer. Consequently, allowances are made in the budget of the company for the capital expenses, including the ones involving the replacement of items which are no longer repaired. Capital expenditures thus normally yield benefit over a long period of time resulting into fixed assets. The resource constraint is a frequent phenomenon of all the economic activities in business. In addition, when a firm is able to spend on specific items it is not willing to do so (Nice, 2002). Therefore, a systematic screening is established to accept or reject the investment proposal. Investment proposal are divided into two groups that is: Mutually exclusive proposals and independent proposals. Mutually exclusive proposals are proposals that have an alternative of doing the same thing. If one alternative is selected then the other one must be rejected for example: if in plant material facilities are required, they are grouped according to their economic benefits. The economic benefits of each of the proposal will be evaluated and the one with the contributing maximum economic benefits is chosen while the rest with less economic benefits are rejected (Pike and Neale, 2003). Ind ependent proposals are those items of capital expenditures that are always considered for different types of projects whose accomplishments are highly needed. In this case all independent proposals are independent of each other and are worthy for implementation. However, due to financial difficulties, priorities are assigned to each proposal according to the gravity of the need of the organization for example: in line with the material handling equipments, instruments such as machines for weighing, packing, stamping may be required(Cotts, 2007). Thus for mutually exclusive proposals the decision criterion is accept or reject while for independent proposals the decision criterion is mainly based in ranking. The decision taken is based on the methods of analyzing the capital budgeting decisions. The environment of capital expenditure proposals are widely grouped into: Expansion, Replacement, Diversification and Strategic proposals. Expansion proposals involve the capital expenditure t o boost the production capacity within the same line of production (Shah, 2007).The investments are basically made in the familiar areas of activity as it involves minimal business risk as compared to diversification, however, larger risk than replacement expenditure. Replacement capital expenditure implies replacement of old machinery by a new one or a modern one. This replacement only

What is Plagiarism Essay Example | Topics and Well Written Essays - 500 words - 12

What is Plagiarism - Essay Example It is even found in articles found in newspapers and magazines; for example in 2010, renowned New York Times reporter and Pulitzer winner were Chris Hedges was found to plagiarize directly from another reporter’s work. At the college level where we are exposed to so many research articles and writings and are often expected to present writings of our own, it becomes almost impossible not to succumb to the temptation of using materials that we may have read or seen somewhere. This is where most students and scholars go wrong. Students are not aware of the fact that even paraphrasing without citation is very much unethical and therefore most end up plagiarizing owing to ignorance of rules of ethical writing. The habit of abiding by plagiarism rules needs to be instilled from a young age. Students need to be first made aware about the existence of â€Å"plagiarism† and its rules and should be first given warnings about plagiarizing in their own papers. They should be encouraged to read articles but also acknowledge them in case they are using the same for their own work, hence students must also be introduced to citation approaches. This initial introduction at least ensures that all students are equally aware of plagiarism and therefore any student found indulging in plagiarism, later on, would have done it out of choice rather than ignorance. Students found to plagiarize should be punished according to the degree of plagiarism detected. The punishment should become harsher with subsequent instances by the same person. Even though homework might seem less important than research articles, students and scholars found to indulge in any kind of plagiarism need to be treated equally a nd hence punishment should be the same for usage of plagiarized material.

Case Study - Business Communication Experience

- Business Communication Experience - Case Study Example In contrast, the two-way management communication system encourages employees to put their maximum potential and avoid under-performance; this approach develops a sense of ownership in employees. In the following parts of the paper, first both management communication systems of two managers have been discussed. It is followed by the segment mentioning more effective manager. After the parts of impact on the communication systems and implications of both approaches, a conclusion has been provided. Management Communication Systems of Two Managers Both managers use different approaches for communication purposes. While working in Unisys Corporation as a Controller in Accounting, I observed that the manager was using the one way management communication approach. It is a form of communication in which a person sends a message to another person without expecting any question, feedback or interaction to follow (Nelson and Quick, 2013, p. 283). In this approach, he did not require my or an y other employee’s feedback. In this approach, it is assumed that managers develop an opinion that his or her message would be adequate and clear to the receptors; they consider that by using a precise and clear language, the decision will be accordingly understood and complied by the receptors. However, my experience in this organization convinces me that this management communication strategy remains considerably less effective and defective as the managers do not take into account the required feedback of their sub-ordinates and other employees and this creates a gap, which leaves unfavourable and less constructive effects on the organizational communication, goals and objectives. In contrast to the above mentioned managerial communication approach, the manager at Maverick Real Estate employs feedback communication approach, known as the circuit communication, which requires that feedback is highly essential for entertaining the objective of effective managerial communicat ion strategy. It is also known as a two-way communication occurs when the receiver extends feedback to the sender (Miller and Braswell, 2011, p.16). Additionally, this approach begins with the presupposition that the manager must know and understand problems that employees are encountering in fulfilling the tasks assigned and their views and opinions relating to their official duties and responsibilities. In addition to that, the manager behaviour remained professional and competent. I did not feel much hesitation and the presence of professional managerial behaviour enabled me to come into contact with the manager and discuss matters relevant to my job description. This scenario was totally absent in the above mentioned previous employment where I remained less comfortable and more fearful due to the less accommodating behaviour shown by the manager at Unisys Corporation. While working as a broker in the Maverick Real Estate, the manager used the intranet for the purpose of effecti ve and prompt communication between and among the staff members. More Effective Manager The manager at the Maverick Real Estate remains more effective in comparison with the manager at the Unisys Corporation. First, the behaviour of the manager created a congenial and employee friendly workplace environment. In which, every employee considers himself or herself to be an important part of the organization. It motivates and increases the level of confidence. With this

Write a paper on the environment and the damages that humans have done Research

Write a on the environment and the damages that humans have done to it - Research Paper Example Human activities lead to different impacts on various components of the ecosystem such as land, biodiversity, and aquatic, terrestrial, marine and the atmosphere. Humans are inherently selfish and want the best for themselves without caring about the impacts of their activities to other members of the ecosystem. Global warming is the most critical problem facing the globe today. Global warming is described as the rise in the average temperature of the earth’s atmosphere and the oceans. This increase was noted to have started in the 19th century and is anticipated to continue rising. Goudie (22) notes that since the beginning of the 20th century, global temperatures have risen by about 0.8o C with approximately two thirds of this increase happening since 1980. Global warming is attributed to increased concentration of greenhouse gases which include carbon dioxide, methane and ozone. However, carbon dioxide is the most significant greenhouse gas. Greenhouse gases lead to global warming by trapping infrared radiations emitted from the earth’s surface in the lower atmosphere thereby causing the temperatures to become warmer. These gases, however, allow radiations from the sun to get to the earth surface but absorb them as they absorb short wave radiations when re-emitted coming fr om the land. Greenhouse gases are important to maintain warm temperatures with the earth surface, but increased concentration of these gases can lead to devastating impacts on the environment, as explained by Goudie (22). Human activities are the most significant causes of increased carbon dioxide levels. Goudie (23) observes that since industrial revolution, the amount of greenhouse gases in the atmosphere has increased significantly. He notes that carbon dioxide and methane concentration has risen by 36 and 148 percent respectively from 1750 levels. Burning of fossil fuels is attributed for more than

The Cross-border Merger of Kraft and Cadbury Term Paper

The Cross-border Merger of Kraft and Cadbury - Term Paper Example An overview of the Kraft –Cadbury merger In February 2010, Cadbury gave in to Kraft’s US$ 19.7 billion takeover after a fierce battle lasting over 100 days. Kraft Foods US is a major confectionary maker. The British chocolate maker had earlier in 2009, rejected a US$ 16.4 billion hostile takeover bid from Kraft, stating that the value did not represent the intrinsic value of the Cadbury brand. Industry experts believe that the combined group is the number one in chocolate and confectionary segments, as well as the second in the high growth gum segment (Ralph & Olesseni, p.61). Cadbury had agreed for 840 pence per share which would give them a total valuation of $19 billion. Media reported that Cadbury slipped into US giant Kraft Foods and the British Prime Minister committed that the jobs in UK could be protected. It was estimated that Cadbury employees numbered more than 45000 worldwide. It was expected, Kraft Cadbury combined would generate large cost savings, enablin g Kraft to become a global market leader. The conglomeration would also generate annual sales of more than $ 50 billion. The market reaction was mixed especially from UK where the fear of job loss came up and cultural reaction was that the country’s honor namely Cadbury’s brand, had been given to US. Kraft Foods was one of the major US confectionery manufacturers with net revenue of $42 billion and operating in 150 countries as of 2008. It was founded 1903 as a cheese company by James L. Kraft (Funding Universe, 2002) and over the years established fine brands like Milka, Toblerone, Jacobs, Oscar Mayer and Oreo. Even though Kraft was able to capture US and European markets, it was the second largest food company in the world and Nestle, Switzerland continued to occupy the premier position with its brands firmly established not only in developed countries but also in developing countries. Nestle had reported a net profit of $9.55 billion with an annual turnover of $99 b illion in 2009. Next in the race for second position was Cadbury, UK with its popular brands like Dairy Milk bars, Roses chocolates, Trident gum and Halls cough drops, built over 150 years not only in UK and developed countries but also firmly established its presence in the developing countries like India, Mexico and Brazil for over 50 years. Cadbury’s revenues in 2008 stood at ?5.4billion. Kraft Foods US with an ambition to reach the top slot in the global confectionery market made a bid for $10 billion to acquire a 100% stake in Cadbury at the end of 2009. The bid was rejected outright as the market value of the share was more than ? 7 per share and Kraft Foods had to reconsider the valuation process of Cadbury and made a revised offer of around $ 19.6 billion in early 2010 over which the shareholders of Cadbury numbering over 90% consented to the acquisition. Evolution and Growth of Kraft Foods Kraft Foods Inc., the second largest food company in the world, had brands spr ead over five consumer sectors – snacks, beverages, cheese, grocery and convenient meals. Kraft Foods had strong presence worldwide and operated in150 countries as of 2008. The company had evolved from a cheese company, started by James L. Kraft in 1903. James L. Kraft had started his cheese business to relieve the grocers from travelling daily to procure cheese. The merger of Kraft – Phenix and National Dairy Products Corporation in 1930 led to the further growth of Kraft. New brands such as Miracle Whip salad dressing, Velveeta pasteurized process cheese spread, were launched and turned to be successful. Innovative advertising strategy followed by Kraft was another driving force for Kraft’s success. The company was renamed as Kraft Foods Company in 1945 and during the post war period Kraft Foods continued with its new product launches and innovative advertising. In spite of various restructuring activities, Kraft General Foods’ financial results were no t rosy. In early 1995, the three units, Kraft USA, General

MRSA Infection Research Paper Example | Topics and Well Written Essays - 2000 words

MRSA Infection - Research Paper Example Antibiotics used to treat ordinary S. aureus infections are rendered useless in case of MRSA. Infection occurring in healthcare settings is called health care-associated MRSA (HA-MRSA) while that occurring in the community, among healthy individuals, is called community-associated MRSA (CA-MRSA) (mayoclinic.org 1). HA-MRSA infections are generally acquired through invasive procedures or clinical devices such as artificial joints, surgeries, intravenous tubings, and catheters while CA-MRSA generally spreads through skin contact among child care workers, high school wrestlers and people living in crowded areas (mayoclinic.org 1). MRSA infection occurs in various parts of the body and owing to its antibiotic resistance, it is difficult to treat. Mild infections result in boils and sores on the skin. MRSA can also infect lungs, urinary tract, and bloodstream (webmd.com 1). There has been an alarming spread in the incidence of tough MRSA strains and because of its antibiotic resistance, MRSA is also called superbug (webmd.com 1). As per CDC, less than 2% of the US population carries MRSA (mayoclinic.org 3). MRSA was discovered in 1961 and research has shown that, like ordinary strains of S. aureus, it is also carried by many healthy people on their bodies, especially their noses (Matheson et al. 299). MRSA infection is common among those with a weaker immune system and while it is commonly a hospital-acquired the infection, its incidence in people who have not been hospitalized has become recently significant (webmd.com 1).

Causes of Salem wWtch Trials Research Paper Example | Topics and Well Written Essays - 750 words

Causes of Salem wWtch Trials - Research Paper Example There was an influx of refugees in Salem village because of war outbreak in 1689, between the English rulers (William and Mary) and France. This made life too hard in this village, as people struggled with the available scarce resources for survival (Blumberg, 2007). In addition to this, the villagers faced diseases, harsh winters and crop failure. The Puritans believed that good fortunes always came from God and were a blessing to them, while bad fortunes were associated with the devil’s work. These people believed that witches were people who had deals with the devil and received powers from the devil in return, for doing evil. According to Puritans, a convicted witch was sentenced to death because it was believed that they could destroy communities and corrupt good Christian people. Although the Puritans had over the years believed in witches, everything changed in 1692, when witch hunt widely spread for the first time. The village of Salem was the centre for accusation. Th is was after two girls, Betty Parris who was 9 years old, the daughter of Reverend Samuel Parris (the first ordained minister in the village), and his niece, Abigail Williams 11 years old accused three women of casting spells on them. Two of the women were Puritan women and the other one was a slave woman. The two girls suffered from a strange sickness, acted oddly, had incomprehensible speech and their bodies were twisted from their original positions into uncomfortable ones. When the girls were diagnosed, there was no reasonable diagnosis found. The doctors tried to search in their medical books but it was unsuccessful. This worried the villagers and made them search desperately for an explanation. It was then that it was concluded that the girls were under the spell of witchcraft by their fellow community members (Yolen and Stemple, 2004). The three women were arrested on February 29 and more than 150 other â€Å"witches† were also arrested and put on trial. By late Septem ber 1692, some had already been put to death and more others died while in jail. During this period, people fasted and prayed for the girls for God’s intervention but it did not succeed. However, although witchcraft began in Salem village making it very famous in rounding up accused witches, the fear of witchcraft increased over the following year. This made the life there more difficult with neighbours rising against their fellow neighbours as others tried to prove the innocence of their dear ones, the accusers worried of what would befall them while the leaders struggled to understand the happenings (Doeden, 2011). In early 1970s, psychologist Linnda Caporael, now a behavioural psychologist at New York's Rensselaer Polytechnic Institute, began to investigate the Salem Witch trials while still a college student with no idea that a common grain fungus could have been the cause of the 1962 events. In 1976, he came up with a theory, which believed that a certain type of food po isoning called convulsive ergotism might have been responsible for the girls’ condition. Convulsive ergotism occurs when a person consumes rye crop- wheat containing a mould called Ergot, which was used to make bread. This causes hallucinations, vomiting, crawling sensations on the skin among many other symptoms similar to those reported in Salem witchcraft trials. It was also discovered that, ergot thrives in damp, rainy springs,

City of Mount Rainier Essay Example for Free

City of Mount Rainier Essay Neither does the department of Economic Development for the City of Mount Rainier handle nor is it involved with any matrix of the pricing strategy. As the Organization for Economic Co-operation and Development (OECD) in 2006 stated on production, packaging and pricing, production cycle involves a flow from the dealers, employees and customers automatically between the functional areas and internationally. Reduced ‘product to market’ cycle meant to ensure full and on time delivery to customers keeps a business on track by reducing errors, inefficiencies and high running costs. However, the City does offer a mix of products solely based on promoting development of blighted or under used parcels. Thus, it is critical that any incentive or business assistance be directed to accomplish the marketing objectives of the City of Mount Rainier. For example, the city will offer or sponsor Tax Increment Financing, State grants or Infrastructure Grants to developers to increase interest and make the area more attractive. This is very necessary for the City to be able to compete with other local municipalities and the District of Columbia, making it imperative for the City to use incentives for redevelopment of the downtown business district. Especially considering that the addition of businesses will create higher paying jobs, large capital investment, tourism and greater benefits to the local community. With this in mind, its primary objective is to add to the diversity of the current business population, increase essential tax base in the area, and pursue the highest quality of jobs and investment by providing an overall balanced approach to economic development for the City. Organizations channel design. A good organizations’ channel design enables exchange of information and ideas with external or outsourced personnel such as financial assistants, engineers, designers, and other partners. It is a system that provides solutions including help to speed up development processes and enhance further future improvements in efficiency With this the City of Mount Rainier can be able to streamline the entire value chain, including all key suppliers, subcontractors, and service providers. The system platform raises transparency of production processes, permitting them to identify potential problems at an early stage, and to take immediate action. According to Kotler and Keller (2008), â€Å" a marketing channel performs the work of moving products from producers to consumers, overcoming the time, place, and possession gaps that separate goods and services from those that want them† (p. 232). The Department of Economic Development for the City of Mount Rainier employs a two –level channel of marketing with at least two intermediaries. The City’s intermediately roles are that of an agent and facilitator. Attracting entrepreneurs and private investment opportunities is the primary goal of economic development in the City of Mount Rainier. As for the Department of Economic Development, the greatest aspiration is to turn around Mount Rainier, making the City one of the best places in the world in which to start a profession or grow a business. The department is working to reposition Mount Rainier as the next great American city for business investment and economic opportunity. This entails employing a balanced attack with corresponding efforts to improve business development, while pursuing transformative public policy changes planned to enhance the magnetism of Mount Rainier as a place in which to invest. On the other hand, an integrated logistics system consists of materials management, material flow systems, and although the City of Mount Rainier does not use such a system as a tool for economic development, A fully integrated accurate unit tracking system is a critical component of modern customer relations and service that is in use, and it is also important for inventory and cost control. Just like integrated logistics system, it monitors all the key movements of goods and services in any transaction. Tracking of the way financial agreements are facilitated for business assistance is important because the gathered data is useful for financial analysis. In support of business analysis, the system ought to have a tool that provides the reporting intelligently. (OECD, 2006) Business information across all assistants can be automatically converted into business intelligence reports benchmarking the performance of the business transactions. At the same time analysis on dealerships can show the best business practices for individual or the group of partners. Marketing and communication strategy Considering that City of Mount Rainier is recognizing a need for business retention and attraction efforts, they have partnered with other public and private businesses stakeholders to create an Economic Development Corporation (EDC) to promote the City and its business advantages and opportunities to local, regional, national and international markets. The purpose of the EDC is to enhance the economic vitality of the City and its residents through creating marketing initiatives, business retention programs, business attraction efforts and dissemination of information. Created as a public-private partnership, the EDC can generate business leads and opportunities that the City could otherwise not do, furthering development. In addition, this partnership serves as the first line of contact with the public, business prospects and developers who are seeking information about doing business in the City. Furthermore, the EDC facilitates development, cultivates business opportunities and creates partnerships that will result in increased economic development in the City of Mount Rainier. The question remains, which supreme marketing and communication strategy would boost this collaboration for better growth? For any company to prosper in any of its transactions, a strong and optimal management system ought to be put in place. In line with Poirier and Reiter, (1996), success in a company is enhanced by rapid and reliable exchange of data within its group members and with its business partners. This is vital to the further development products and deliverance of quality services. Technically one important communication strategy would calls for a highly effective communications infrastructure that allows effective coordination and management of highly complex, highly flexible development, design and production processes. In line with Poirier and Reiter, on Advanced Planning and Optimization in (1996), the main aims of marketing managers should be to process aftermarket into one system, plan for wide coverage and include all parties involved in the planning process. P. 160) In other words, high-speed data communications have to be one of the engines driving the growth of any organization or company offering services or dealing with businesses stakeholders. Any marketing strategy should take establishment and enhancement of a quality communications infrastructure in it key aims. This is a reliable system that ought to augment cooperation among virtual elements regardless of their position at the time of requirement. It permits all around the globe contribution to projects without duplication or underutilization of human effort or technical resources. Consideration for one of the various information systems and networks is a standard solution that can be tailored to particular needs. A system that supports all the data and allows for data interchange with outside suppliers and other partners naturally always gives a good return on investment. This is not a matter of just laying down the network system within the company. Efficiency, Security, reliability and scalability of the system are some of the vital parts to be fulfilled by a system so as to support diverse access methods and service levels. As Poirier and Reiter emphasizes, a good system is one that can be geared to the needs of each individual supplier and business partner. (P. 160). Needless to say, this new ways of revitalizing the City’s commercial corridors by encouraging continued and private investment, would highly revitalize strategies allowing the city to target its assistance and support in ways that are sustainable at the local level.

Pollution in the USA

Pollution in the USA One of the most eminent problems is pollution and other forms of environmental worsening. While this problem is everything but new, and in reality tends to weaken in the most urbanized countries, the innovation is its global growth, leading to such problems as the global warming. The USA is the worlds leading polluter. The USA has the largest cars and the largest roads to all other countries. This is an inconceivable occurrence to the rest of the world, where car accumulation and gasoline use are instantly equated with pollution and capriciousness. In Europe, knowledge is raised by most governments of the pollution and side effects of burning fuels and heavy taxes are obligatory on these in order to furnish public transportation, dead set against pollution solutions and substitute fuel methods such as LPG and electric power-driven cars. Can the USAs government really maintain to hold its version of dictatorship and free enterprise at the expenditure of every human on the planet? The American Government under President G. Bush was just about put in power solely by money from huge oil companies, but something needs to change. The world sees our restriction of pollutant emissions vital and required that we do this. The USA seems to think that it is not viable and expensive and isnt going to do it! We must reduce pollution, it doesnt matter how expensive it is. It has to be done. The world sees Americas unwillingness with repugnance and dismay. The USA has some environmental issues sorted, like domestic waste discarding. During the 1970s and 1980s the USA made itself obligatory in the campaigns to prohibit the discharge of ozone gases; thanks to its help the hole in the ozone layer is gradually closing, finally. The USA polices the worlds military nations, keeping check on their artillery and intentions. It does not do the same with matters of universal health or pollution. The USA is by far the worlds leading polluter and is also the country that is seen as slightest vigorous in combating world pollution. Failure to approve the Kyoto Protocol is a serious error and much of the world is left in astonish and dismay that the USA ignores these problems. Comprising over 70% of the Earths exterior, water is unquestionably the most valuable natural resource that exists on our planet. Without the seemingly very useful mix comprised of hydrogen and oxygen, life on Earth would be absent: it is vital for everything on our planet to nurture and flourish. Even though we as humans know this fact, we ignore it by polluting our rivers, lakes, and oceans. Then, we are slowly but surely harming our planet to the point where organisms are dying at an incredibly frightening rate. As well as to blameless organisms dying off, our drinking water has become significantly exaggerated as is our ability to use water for leisure purposes. In order to fight water pollution, we must be aware of the problems and become part of the solution. According to the American College Dictionary, pollution is defined as: to make foul or unclean; dirty. Water pollution occurs when a body of water is negatively affected due to the accumulation of large amounts of resources to the water. When it is out of condition for its intentional use, water is considered polluted. There are two types of water pollutants that exist; point source and nonpoint source. Point sources of pollution take place when harmful substances are emitted straight into a body of water. The Exxon Valdez oil spill best illustrates point source water pollution. A nonpoint source delivers pollutants in some way through environmental changes. An example of the type of water pollution is when fertilizer from a field is passed into a stream by rain, in the outward appearance of run-off which in turn affects aquatic life. The equipment exists for point sources of pollution to be monitored and synchronized, even though political factors may make matters worse matters. Non point sources are much more complex to control. Pollution that arises from nonpoint sources accounts for a greater part of the contaminants in steams and lakes. There are many causes of pollution, some including, but not limited to, sewage and fertilizers that contain nutrients such as nitrates and phosphates. In surplus levels, nutrients over encourage the growth of aquatic plants and algae. Extreme growth of these types of organisms as a result clogs our waterways, use up dissolved oxygen as they decay, and obstruct light to deeper waters. This in turn, proves very harmful aquatic organisms as it affects the respiration capability or fish and other invertebrates that dwell in water. Pollution is also caused when slit and other poised solids, such as soil, wash off plowed fields, construction and logging sites, urban areas, and eroded river banks when it rains. Pollution in the form of organic objects enters waterways in many unusual ways, such as sewage, as leaves and grass clippings, or as runoff from farm animals feedlots and pastures. Pathogens are another type of pollution that proves very harmful. They can cause many illnesses that vary from typhoid and dysentery to slight respiratory and skin diseases. These pollutants come in waterways through unprocessed sewage, storm drains, septic tanks, runoff from farms, and mainly boats that dump sewage. Despite the fact that they are microscopic, these pollutants have an incredible affect evidenced by their capability to cause sickness. Three preceding forms of water pollution exist in the forms of petroleum, radioactive substances, and heat. Petroleum frequently pollutes water bodies in the form of oil, ensuing from oil spills. These significant unintentional discharges of petroleum are a vital cause of pollution alongside shore lines. In addition to the supertankers, off-shore drilling operations add a large share of pollution. One educated guess is that one ton of oil is spilled for every million tons of oil transported. Its equal to about 0.0001 percent. Radioactive substances are formed in the type of waste from nuclear power plants, and from the industrial, medical, and scientific use of radioactive supplies. Precise forms of waste are uranium and thorium removal and decontamination. The final form of water pollution is heat. Heat is a pollutant because of the raise in temperatures, which cause death in many aquatic organisms. These decreases in temperatures are caused when a release of cooling water by factor ies and power plants take place. Oil pollution is an increasing problem, mainly devastating to coastal wildlife. Small quantities of oil extend hastily across long distances to form deadly oil slicks. The chief sources of water pollution can be classified as municipal, industrial, and agricultural. Municipal water pollution consists of waste water from homes and business establishments. For several years, the most important goal for treating municipal wastewater was simply to diminish its substance of suspended solids, oxygen-demanding materials, dissolved inorganic compounds, and harmful bacteria. In current years, nevertheless, more stress has been placed on humanizing means of discarding of the solid residues from the municipal management process. The important methods of treating municipal wastewater fall into three stages: primary treatment, as well as grit removal, screening, grinding, and sedimentation; secondary treatment, which entails corrosion of dissolved organic matter by resources of using biologically active sludge, which is then filtered off; and tertiary treatment, in which complex biological methods of nitrogen removal and chemical and physical methods such as granular filtration and activated carbon assimilation are working. The management and removal of solid residues can account for 25 to 50 percent of the funds and operational costs of a management plant. The distinctiveness of industrial waste waters can fluctuate significantly both within and among industries. The shock of industrial discharges depends not only on their combined characteristics, such as biochemical oxygen demand and the amount of suspended solids, but also on their substance of specific inorganic and organic substances. Three options are accessible in controlling industrial wastewater. Control can take place at the point of cohort in the plant; wastewater can be pretreated for expulsion to municipal treatment sources; or wastewater can be treated entirely at the plant and either reused or discharged str aight into receiving waters. Agriculture, as well as profitable livestock and poultry farming, is the starting place of many organic and inorganic pollutants in surface waters and groundwater. These contaminants take account of both residues from wearing away cropland and compounds of phosphorus and nitrogen that somewhat originate in animal wastes and viable fertilizers. Animal wastes are high in oxygen challenging material, nitrogen and phosphorus, and they over and over again harbor pathogenic organisms. Wastes from viable feeders are controlled and predisposed of on land; their main threat to natural waters, as a result, is from runoff and leakage. Control many entail settling basins for liquids, some degree of biological treatment in aerobic or anaerobic lagoons, and a range of other methods. Ninety-five percent of all fresh water on earth is ground water. Ground water is established in natural rock formations. These formations, called aquifers, are vital natural resources with many uses. Nationally, fifty-three percent of the inhabitants rely on ground water as a supply of drinking water. In rural areas this number is even more elevated. Eighty-one percent of the society water is reliant on ground water. Although the 1992 Section 305(b) State Water Quality Reports indicate that, overall, the Nations ground water quality is good to excellent; many local areas have experienced significant ground water contamination. Some examples are leaking underground storage tanks and municipal landfills. Without a doubt, the problems connected with water pollution have the capability to disturb life on our planet to an enormous extent. An independent governments purpose is to protect people, its populace, and the populace of the world. In industrial countries it is the Governments job to keep commercialism in order and protect people from disparity, domination and worldly practices of big business. Some people have been required to wonder if the American government can still be considered to be working for the welfare of the people anywhere in the world, or if it is without a doubt merely the worlds largest corporation.

Sperm Assessment Using Flow Cytometry

Sperm Assessment Using Flow Cytometry State of the art in sperm assessment using flow cytometry Abstract Flow cytometry is emerging as an important tool in the field of modern andrology for routine analysis of spermatozoa. Recently, application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, semen sample analysis was routinely performed by microscopical evaluation and manual techniques by laboratory operators; the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. The last decade, several new flow cytometric techniques have been introduced for farm animal semen assessment that enable a more detailed evaluation of several sperm characteristics. Here in this paper, an initiative has been taken to focus on a number of recent flow cytometry developments important for addressing questions in andrological tests. After the invention of flow cytometry, sperm evaluation by traditional microscopic means became questioned due to the robust advantages of flow cytometry over the microscopic method. Due to the recent development of large number of fluroscence probes, flow cytometry is now capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gamets based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the possibility of working with small sample sizes, increases the repeatability of assessment, removes the subjectivity of assessment and allows simultaneous assessment of multiple fluorochromes. Flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of experiments that a re not possible with any other technique. Nowadays, semen evaluation using laboratory assays is extremely important to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen parameters that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Semen evaluation is the single most important laboratory test that has helped us to identify clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Methods are available that can sometimes estimate the potential fertilizing capacity of a semen sample and, in some cases, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005). The methods routinely used for evaluation of the quality of a semen sample involved an evaluation of general appearance (i.e. colour, contamination, etc.), volume, pH, sperm concentration, viability, morphology and motility. Most of these techniques are microscopic analyses that only measure a small number of spermatozoa within a population, are time-consuming, can be subjective and generally measure sperm attributes individually. Recently, limitations of semen evaluation methodology have been brought into sharp focus by controversies raised in the epidemiological literature. It should also be noted that such conventional measurements are prone to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed se men, and IVF. Such handling implies semen extension, fluorophore loading, ultraviolet and laser illumination, high-speed sorting, cooling and cryopreservation, procedures that impose different degrees of change in sperm function following damage to sperm membranes, organelles or the DNA. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed that buck of these procedures are incomplete, time consuming and laborious. Flow cytometry in different technical applications offers many advantages for the analysis of sperm quality. Flow cytometry allows the simultaneous measurement of multiple fluorescences and light scatter induced by illumination of single cell or microscopic particles in suspension, as they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers can acquire data on several subpopulations within a sample in a few minutes, making it ideal for assessment of heterogenous populations in semen sample. Initially developed in the 1960s, flow cytometry made automated separation of cells based on the unique recognition of cellular patterns within a population feasible (Hulett et al., 1969). Using such a separation approach, cellular patterns can be identified by as sessing, in individual cells within a population, protein expression using fluorescently labeled antibodies and other fluorescent probes (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). Flow cytometry was first developed for medical and clinical applications such as haematology and oncology. These areas still account for the vast majority of publications on this technique, but during the past few years it has been used in other areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. FCM is increasingly used for basic, clinical, biotechnological, and environmental studies of biochemical relevance. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream. In other word, a flow cytometer will be defined as ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other work, cytometry refers to the measurement of physical and/or chemical characteristics of cells or, by extension, of other particles. It is a process in which such measurements are made while the cells or particles pass, preferably in single file, through the measuring apparatus in a fluid stream. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other analytical tools, where a single value for each parameter is obtained for the whole population, flow cytometry provides data for every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry gives objective and accurate results (Bunthof et al., 2001; Shleeva et al., 2002), overcoming the problems with the manual methods described above. Function and types of flow cytometry Fluidics, optics and electronics are the three main systems that make up a flow cytometer. In a few minutes, the flow cytometer can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment began when it was used for measuring their DNA content (Evenson et al., 1980) and its application to semen analysis has gradually increased over the last 10-15 years. Flow cytometry is now applied to semen evaluation of traits such as cell viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity and DNA status. New fluorescent stains and techniques are continuously being developed that have potential application to the flow cytometric evaluation of spermatozoa. Flow cytometry permits the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and the more sperm parameters that can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters. Sorters have the ability not only to collect data on cells (analyse cells) but also to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function During the past 2 decades, there has been an increasing interest in reliable assays for assessing semen quality in the fertility clinic and artificial insemination industries. The use of flow cytometry for sperm analysis is an attempt to address the long-standing problem of the subjective nature of the manual method commonly used for semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with manual techniques. Because of time and cost restraints, most laboratories analyze only 50 to 100 sperm to compute the percentage of each cell population and the viability rate. This small sample from a population of millions probably results in a statistical sampling error (Russel and Curtis, 1993). The conventional methods used are limited to microscopic determination of sperm concentration using a hemocytometer (Jorgensen et al., 1997) and evaluation of sperm motility and morphology (Keel et al., 2002). These methods usually involve a subjective asses sment of a few hundred sperm, and quality assurance is rarely implemented in the laboratories performing such analysis. Flow cytometry is a technique that is superior to conventional light microscopy techniques in terms of objectivity, number of cells measured, speed, and precision (Spano and Evenson, 1993). The technique has been used on human sperm to determine a number of factors, including membrane integrity, mitochondrial function, acrosome status, and multiparameter measurement (Garrido et al., 2002). Flow cytometry permitted us to analyze thousands of cells in few seconds. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in diverse species (boar, bull, stallion etc), and the observed CVs were significantly better than those reported for the manual method. While there are many advantages of using the flow cytometer for routine semen analysis, its use is often limited to research by the expense and difficulties of operation associated with the requirement of a skilled operator. In addition, a flow cytometer is quite large and cannot withstand shocks associated with movement, meaning it requires a dedicated position in the laboratory. However, the development of more affordable ‘‘bench-top flow cytometers has recently increased the potential application to semen analysis. If we consider flow cytometric analysis further, we can see that it is gaining wider acceptance as a technique for assessing the acrosome reaction and viability simultaneously. Comparing these assays to the more widely used epifluorescent microscopic techniques, the flow cytometric analysis is able to give a far more simple and objective method of analysis, especially with regard to correlation of fertilization with acrosome reactivity potential (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Preliminary data from a number of studies suggested that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). Using this information, the World Health Organization stated that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, may lack the accuracy and precision of the haemocytometer technique (World Health Organization, 1999). Further data—for example, from Tomlinson and colleagues—showed that 2 proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) gave lower spe rm concentrations compared to the hemocytometer method (Tomlinson et al., 2001). To put this in context, numerous reports document unacceptable discrepancies between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Several reports emphasize the need for improvement in overall quality of semen testing within and between laboratories (Neuwinger et al., 1990; Jorgensen et al., 1997; Keel et al., 2000). However, the subjective nature of conventional semen analyses, combined with their relatively low precision due to the low number of cells assessed, leads to poor intra- and interlaboratory reproducibility; therefore, the introduction of standardized or quality controlled procedures will probably have a limited effect. The conventional analyses are used to determine whether parameters obtained from an ejaculate are within the range characterized by fertile men, and these methods can therefore provide only unclear cut-off values when used for the prediction of fertility status. Many of the advantages that accrue when using flow cytometry may, when applied to assessment of sperm cells, help overcome some of the mentioned problems found in conventional semen analysis. In the field of semen analysis, validation of a method is important because it is essential to have specific, precise, objective, and accurate laboratory tests to establish a correlation of the data with fertility or to determine the fertility potential of a semen sample correctly (Amann, 1989). Precision of a laboratory test is of great concern to the andrologist in the fertility clinic, since the results of the semen analysis are often used to advise a patient about his fertility and the prognosis for the treatment of the couple. To use established cut-off values and ensure uniform diagnosis, within and between laboratory variations should be determined and followed closely. Accurate determination of sperm cell concentration is critical to the AI industry because it provides assurance both to bull studs and to customers that straws of extended semen contain the sperm numbers indicated. An accurate measure of sperm concentration is particularly important in export markets in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Hemacytometers are widely used for routine sperm counts, but the equipment is slow, and multiple measurements of each sample are needed. Single hemacytometer counts are not highly accurate; because of inherent errors in the technique, Freund and Carol (13) found that mean differences of 20% were not uncommon between duplicate sperm count determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (19). Currently, the primary method used by the AI industry to estimate sperm concentration is spectrophotometric determination of turbidity of a semen sample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (1). This approach is only as accurate as the methods used for spectrophotometer calibration. New, more accurate methods for sperm count determinations are being sought to replace the older ones. Some laboratories are trying the Maklerm counting chamber (Seif- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. It may be argued that when comparing fluorescent microscopy assays with flow cytometry, one is examining patterns of fluorescence rather than fluorescence intensity, i.e., the flow cytometer is not capable of discriminating sperm which have a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is no significant difference between the two methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Viability The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Currently flow cytometric procedures have been developed which simultaneously evaluate sperm cell viability, acrosomal integrity and mitochondrial function. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 1997), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 (R123) to assess mitochondrial membrane potential and ethidium bromide to determine membrane integrity using flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). At present, one of the most commonly used viability stain combinations is SYBR-14 and PI, sold commercially as LIVE/DEAD Sperm Viability kit (Molecular Probes Inc., OR, USA). When used in combination, the nuclei of living sperm fluoresce green (SYBR-14) and cells that have lost their membrane integrity stain red (PI). This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been used to evaluate boar sperm concentration [11]. Due to its compact size and its relatively inexpensive purchase price, this instrument could be useful for field measurements of both concentration and viability. This instrument was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and found a negative correlation between the percentage of damaged cells and field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by either approach can be used in combination with each other. For example, when CFDA is used along with PI, three populations of cells can be identified: live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa. Almlid and Johnson (1988) found this combination useful for monitoring membrane damage in frozen-thawed boar spermatozoa during evaluation of various freezing protocols. Harrison and Vickers (1990) also used this combination with a fluorescent microscope and found it to be an effective indicator of the viability of fresh, incubated or cold-shocked boar and ram spermatozoa. Garner et al. (1986) used this combination to stain spermatozoa from a number of species, but at that time could not find a relationship between bull sperm viability detected by CFDA/PI and fertility. Flow cytometry for assessment of sperm viability appears to be a valuable tool for the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect on NRR56. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combination of assessment of sperm viability and concentration appears to be useful in the improvement of quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. The mechanism by which SYBR-14 binds to the DNA is not known. It is know that PI stains nucleic acids by intercalating betwee n the base pairs (Krishan, 1975). Viability stains have also been used in association with fluorescently labeled plant lectins to simultaneously assess the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). Assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) demonstrated that insemination of boar spermatozoa stained with SYBR-14 into sows did not compromise fertilization or the development of flushed porcine embryos in culture. Non-viable cells can be determined using membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. An intact plasma membrane will prevent these products from entering the spermatozoa and staining the nucleus. Commonly used examples include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). Wilhelm et al. (1996) compared the fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality and found that viability, as assessed by flow cytometry using PI, was the single laboratory assay that correlated with stallion fertility. Changes in sperm membrane permeability Detection of increased membrane permeability is employed in different cell types to distinguish different status of membrane organization (Cohen, 1993; Ormerod et al., 1993; Castaneda and Kinne, 2000; Reber et al., 2002). Sperm plasma membrane status is of utmost importance due to its role, not only as a cell boundary, but also for its need for cell-to-cell interactions, e.g. between spermatozoa and the epithelium of the female genital tract and between the spermatozoon and the oocyte and its vestments (for review, see Rodriguez-Martinez, 2001). Membrane integrity and the stability of its semipermeable features are prerequisites for the viability of the spermatozoon (Rodriguez-Martinez, 2006). However, cryopreservation, whose purpose is to warrant sperm survival, causes irreversible damage to the plasma membrane leading to cell death in a large number of spermatozoa (Holt, 2000) or, in the surviving spermatozoa, to changes similar to those seen during sperm capacitation, thus shorten ing their lifetime (Perez et al., 1996; Cormier et al., 1997; Maxwell and Johnson, 1997; Green and Watson, 2000; Schembri et al., 2000; Watson, 2000). During the freezing process, cells shrink again when cooling rates are slow enough to prevent intracellular ice formation as growing extracellular ice concentrates the solutes in the diminishing volume of non-frozen water, causing intracellular water exosmosis. Though warming and thawing, the cells return to their normal volume. Thus, it is important to know the permeability coefficient of the cells to cryoprotectants, as well as the effect of cryoprotective agents on the membrane hydraulic conductivity. Classical combination of probes allows discrimination of two or three subpopulations of spermatozoa, i.e. live, dead and damaged depending on the degree of membrane integrity (Eriksson RodrÄ ±Ã‚ ´guez-MartÄ ±Ã‚ ´nez, 2000). A new, simple and repeatable method to detect membrane changes in all spermatozoa present in a boar semen sample, by use of markers (combination of SNARF-1, YO-PRO-1 and ethidium homodimer) used to track changes in sperm membrane permeability, has been developed recently by our group (Pena et al., 2005). In determined physiological or pathological situations, live cells are unable to exclude YO-PRO-1, but are still not permeable to other dead-cell discriminatory dyes, like propidium iodide or ethidium homodimer. YO-PRO-1 is an impermeable membrane probe and can leak in, only after destabilization of the membrane, under conditions where ethidium homodimer does not. Because several ATP-dependent channels have been detected in spermatozoa (Acevedo et al. , Sperm Assessment Using Flow Cytometry Sperm Assessment Using Flow Cytometry Abstract Flow cytometry is emerging as an important tool in the field of modern andrology for routine analysis of spermatozoa. Recently, application of flow cytometry in the artificial insemination industry especially for pig is a new approach. Until very recent, semen sample analysis was routinely performed by microscopical evaluation and manual techniques by laboratory operators; the analysis is affected by a wide imprecision related to variability among observers, influencing its clinical validity. The last decade, several new flow cytometric techniques have been introduced for farm animal semen assessment that enable a more detailed evaluation of several sperm characteristics. Here in this paper, an initiative has been taken to focus on a number of recent flow cytometry developments important for addressing questions in andrological tests. After the invention of flow cytometry, sperm evaluation by traditional microscopic means became questioned due to the robust advantages of flow cytometry over the microscopic method. Due to the recent development of large number of fluroscence probes, flow cytometry is now capable of analyzing number of sperm characteristics like viability, capacitation, acrosomal integrity, membrane permeability, membrane integrity, mitochondrial status, DNA integrity, decondensation of DNA and differences between gamets based on sex. The application of flow cytometry to their detection allows increased numbers of spermatozoa to be assessed over a short time-period, provides the possibility of working with small sample sizes, increases the repeatability of assessment, removes the subjectivity of assessment and allows simultaneous assessment of multiple fluorochromes. Flow cytometry is a technique capable of generating significantly novel data and allows the design and execution of experiments that a re not possible with any other technique. Nowadays, semen evaluation using laboratory assays is extremely important to the artificial insemination industry to provide the most desired quality product to customers. Future development of flow cytometric techniques will permit further advances both in our knowledge and in the improvement of assisted reproduction techniques. In this paper, the main semen parameters that can be analyzed with fluorochromes and adapted for use with a flow cytometer will be reviewed and the relationship of these tests to fertility will be discussed. Introduction Semen evaluation is the single most important laboratory test that has helped us to identify clear-cut cases of fertility (Jarow et al., 2002), infertility or even of potential sub-fertility (Rodrà ­guez-Martà ­nez, 2007). Determination of the potential fertility of semen sample and, in the long run, of the male from which it has been collected is the ultimate goal of semen evaluations in clinically healthy sires. Methods are available that can sometimes estimate the potential fertilizing capacity of a semen sample and, in some cases, of the male (reviewed by Dziuk 1996; Rodrà ­guez-Martà ­nez et al. 1997a; Rodrà ­guez-Martà ­nez and Larsson 1998; Saacke et al. 1998; Larsson and Rodrà ­guez-Martà ­nez 2000; Rodrà ­guez- Martà ­nez 2000, 2003; Popwell and Flowers 2004; Graham and Mocà © 2005; Gillan et al. 2005). The methods routinely used for evaluation of the quality of a semen sample involved an evaluation of general appearance (i.e. colour, contamination, etc.), volume, pH, sperm concentration, viability, morphology and motility. Most of these techniques are microscopic analyses that only measure a small number of spermatozoa within a population, are time-consuming, can be subjective and generally measure sperm attributes individually. Recently, limitations of semen evaluation methodology have been brought into sharp focus by controversies raised in the epidemiological literature. It should also be noted that such conventional measurements are prone to extreme inter-ejaculate variation, even when the laboratory methodology has been standardized. In the wake of this information, new opportunities have arisen for the development of methods for the diagnosis of male infertility, many of which have been shown to exhibit a prognostic value that eludes conventional semen profiling. Moreover, ejaculated spermatozoa are nowadays handled for use in assisted reproductive technologies, such as the artificial insemination of chilled, frozen-thawed or sexed se men, and IVF. Such handling implies semen extension, fluorophore loading, ultraviolet and laser illumination, high-speed sorting, cooling and cryopreservation, procedures that impose different degrees of change in sperm function following damage to sperm membranes, organelles or the DNA. Therefore, although several assays have been developed to monitor these sperm parameters, recently it is being claimed that buck of these procedures are incomplete, time consuming and laborious. Flow cytometry in different technical applications offers many advantages for the analysis of sperm quality. Flow cytometry allows the simultaneous measurement of multiple fluorescences and light scatter induced by illumination of single cell or microscopic particles in suspension, as they flow very rapidly through a sensing area. The increasing use over the past decade of flow cytometry in the leading laboratories in human and veterinary andrology has dramatically increased our knowledge of sperm function under physiological and biotechnological conditions. Flow cytometers can acquire data on several subpopulations within a sample in a few minutes, making it ideal for assessment of heterogenous populations in semen sample. Initially developed in the 1960s, flow cytometry made automated separation of cells based on the unique recognition of cellular patterns within a population feasible (Hulett et al., 1969). Using such a separation approach, cellular patterns can be identified by as sessing, in individual cells within a population, protein expression using fluorescently labeled antibodies and other fluorescent probes (Baumgarth and Roederer, 2000; Herzenberg et al., 2006). Flow cytometry was first developed for medical and clinical applications such as haematology and oncology. These areas still account for the vast majority of publications on this technique, but during the past few years it has been used in other areas, such as bioprocess monitoring, pharmacology, toxicology, environmental sciences, bacteriology and virology. Recent advancement of flow cytometry increased its application in the reproductive biology especially for andrology. FCM is increasingly used for basic, clinical, biotechnological, and environmental studies of biochemical relevance. Although flow cytometry may overestimate the population of unlabelled cells (Petrunkina and Harrison, 2009), plethora of research from our group in pig (Pena et al., 2003, 2004, 2005; Spjuth et al., 2007; Fernando et al., 2003; Saravia et al.,2005, 2007,2009; De Ambrogi et al., 2006; ) bull (Bergquist et al., 2007; Nagy et al., 2004; Januskauskas et al., 2003; Bergqvist et al., 2007; Hallap et al., 20 05, 2006;) stallion ( Kavak et al., 2003; Morrell et al., 2008) indicate that newly developed fluorescent stains and techniques of flow cytometry has made possible a more widespread analysis of semen quality at a biochemical, ultrastructural and functional level. Therefore, flow cytometry is the current technical solution for rapid, precisely reproducible assessment of sperm suspensions. In this review we have described potentiality and scope of flow cytometry for the evaluation of semen, and the way in which this technique can be used in clinical applications for andrology based on some of our previous experiences. Definition of flow cytometry The definition of a flow cytometer is ‘an instrument which measures the properties of cells in a flowing stream. In other word, a flow cytometer will be defined as ‘an instrument that can measure physical, as well as multi-colour fluorescence properties of cells flowing in a stream. In other work, cytometry refers to the measurement of physical and/or chemical characteristics of cells or, by extension, of other particles. It is a process in which such measurements are made while the cells or particles pass, preferably in single file, through the measuring apparatus in a fluid stream. The data obtained can be used to understand and monitor biological processes and develop new methods and strategies for cell detection and quantification. Compared to other analytical tools, where a single value for each parameter is obtained for the whole population, flow cytometry provides data for every particle detected. As cells differ in their metabolic or physiological states, flow cytometry allows us not only to detect a particular cell type but also to find different subpopulations according to their structural or physiological parameters. Flow cytometry is a technique for measuring components (cells) and the properties of individual cells in liquid suspension. In essence, suspended cells are brought to a detector, one by one, by means of a flow channel. Fluidic devices under laminar flow define the trajectories and velocities that cells traverse across the detector, and fluorescence, absorbance, and light scattering are among the cell properties that can be detected. Flow sorting allows individual cells to be sorted on the basis of their measured properties, and one to three or more global properties of the cell can be measured. Flow cytometers and cell sorters make use of one or more excitation sources and one or two fluorescent dyes to measure and characterize several thousands of cells per second. Flow cytometry gives objective and accurate results (Bunthof et al., 2001; Shleeva et al., 2002), overcoming the problems with the manual methods described above. Function and types of flow cytometry Fluidics, optics and electronics are the three main systems that make up a flow cytometer. In a few minutes, the flow cytometer can acquire data on all subpopulations within a sample, making it ideal for assessment of heterogenous population, such as spermatozoa. The adaptation of flow cytometry to sperm assessment began when it was used for measuring their DNA content (Evenson et al., 1980) and its application to semen analysis has gradually increased over the last 10-15 years. Flow cytometry is now applied to semen evaluation of traits such as cell viability, acrosomal integrity, mitochondrial function, capacitation status, membrane fluidity and DNA status. New fluorescent stains and techniques are continuously being developed that have potential application to the flow cytometric evaluation of spermatozoa. Flow cytometry permits the observation of physical characteristics, such as cell size, shape and internal complexity, and any component or function of the spermatozoon that can be detected by a fluorochrome or fluorescently labeled compound. The analysis is objective, has a high level of experimental repeatability and has the advantage of being able to work with small sample sizes. Flow cytometry also has the capacity to detect labeling by multiple fluorochromes associated with individual spermatozoa, meaning that more than one sperm attribute can be assessed simultaneously. This feature has an added benefit for semen analysis, as few single sperm parameters show significant correlation with fertility in vivo for semen within the acceptable range of normality (Larsson and Rodriguez-Martinez, 2000) and the more sperm parameters that can be tested, the more accurate the fertility prediction becomes (Amman and Hammerstedt, 1993). There are two main types of flow cytometers-analysers and sorters. Sorters have the ability not only to collect data on cells (analyse cells) but also to sort cells with particular properties (defined by the flow cytometer operator) to extremely high purities. There are also a number of commercial flow cytometers that have been developed for particular analytical requirements. Partec manufacture a Ploidy Analyser and also a Cell Counter Analyser. Optoflow has developed a flow cytometer for the rapid detection, characterization and enumeration of microorganisms. Luminex is developing technology for multiplexed analyte quantitation using a combination of microspheres, flow cytometry and high speed digital processing. Advantages of FC compared to other conventional techniques to explore sperm structure and function During the past 2 decades, there has been an increasing interest in reliable assays for assessing semen quality in the fertility clinic and artificial insemination industries. The use of flow cytometry for sperm analysis is an attempt to address the long-standing problem of the subjective nature of the manual method commonly used for semen analysis. An additional source of laboratory variation is the low number of sperms analyzed with manual techniques. Because of time and cost restraints, most laboratories analyze only 50 to 100 sperm to compute the percentage of each cell population and the viability rate. This small sample from a population of millions probably results in a statistical sampling error (Russel and Curtis, 1993). The conventional methods used are limited to microscopic determination of sperm concentration using a hemocytometer (Jorgensen et al., 1997) and evaluation of sperm motility and morphology (Keel et al., 2002). These methods usually involve a subjective asses sment of a few hundred sperm, and quality assurance is rarely implemented in the laboratories performing such analysis. Flow cytometry is a technique that is superior to conventional light microscopy techniques in terms of objectivity, number of cells measured, speed, and precision (Spano and Evenson, 1993). The technique has been used on human sperm to determine a number of factors, including membrane integrity, mitochondrial function, acrosome status, and multiparameter measurement (Garrido et al., 2002). Flow cytometry permitted us to analyze thousands of cells in few seconds. In our series of studies, we demonstrated the feasibility and reproducibility of an automated method to evaluate sperm cell type, count, and viability in human boar samples. In our hand, the precision of the flow cytometric analysis is satisfactory in diverse species (boar, bull, stallion etc), and the observed CVs were significantly better than those reported for the manual method. While there are many advantages of using the flow cytometer for routine semen analysis, its use is often limited to research by the expense and difficulties of operation associated with the requirement of a skilled operator. In addition, a flow cytometer is quite large and cannot withstand shocks associated with movement, meaning it requires a dedicated position in the laboratory. However, the development of more affordable ‘‘bench-top flow cytometers has recently increased the potential application to semen analysis. If we consider flow cytometric analysis further, we can see that it is gaining wider acceptance as a technique for assessing the acrosome reaction and viability simultaneously. Comparing these assays to the more widely used epifluorescent microscopic techniques, the flow cytometric analysis is able to give a far more simple and objective method of analysis, especially with regard to correlation of fertilization with acrosome reactivity potential (Uhler et al., 1993; Purvis et al., 1990; Carver-Ward et al., 1996). A large number of different techniques to estimate sperm concentration have been reported. In the mid-1990s a series of fixed-depth disposable slides were evaluated as rapid and effective pieces of equipment for the estimate of sperm concentration. Preliminary data from a number of studies suggested that, at least in the 20-mm-depth format, such chambers resulted in a noticeable underestimate of sperm concentration compared to the gold standard (improved Neubauer hemocytometer). Using this information, the World Health Organization stated that ‘‘such chambers, whilst convenient in that they can be used without dilution of the specimen, may lack the accuracy and precision of the haemocytometer technique (World Health Organization, 1999). Further data—for example, from Tomlinson and colleagues—showed that 2 proprietary disposable slides (Microcell, Conception Technologies, San Diego, Calif; Leja, Leja Products, BV Nieuw- Vennep, The Netherlands) gave lower spe rm concentrations compared to the hemocytometer method (Tomlinson et al., 2001). To put this in context, numerous reports document unacceptable discrepancies between different laboratories and even between different individuals, although fewer studies attempt to address these issues. So, what is wrong? Several reports emphasize the need for improvement in overall quality of semen testing within and between laboratories (Neuwinger et al., 1990; Jorgensen et al., 1997; Keel et al., 2000). However, the subjective nature of conventional semen analyses, combined with their relatively low precision due to the low number of cells assessed, leads to poor intra- and interlaboratory reproducibility; therefore, the introduction of standardized or quality controlled procedures will probably have a limited effect. The conventional analyses are used to determine whether parameters obtained from an ejaculate are within the range characterized by fertile men, and these methods can therefore provide only unclear cut-off values when used for the prediction of fertility status. Many of the advantages that accrue when using flow cytometry may, when applied to assessment of sperm cells, help overcome some of the mentioned problems found in conventional semen analysis. In the field of semen analysis, validation of a method is important because it is essential to have specific, precise, objective, and accurate laboratory tests to establish a correlation of the data with fertility or to determine the fertility potential of a semen sample correctly (Amann, 1989). Precision of a laboratory test is of great concern to the andrologist in the fertility clinic, since the results of the semen analysis are often used to advise a patient about his fertility and the prognosis for the treatment of the couple. To use established cut-off values and ensure uniform diagnosis, within and between laboratory variations should be determined and followed closely. Accurate determination of sperm cell concentration is critical to the AI industry because it provides assurance both to bull studs and to customers that straws of extended semen contain the sperm numbers indicated. An accurate measure of sperm concentration is particularly important in export markets in which verification of numbers may be required. Routine sperm counts can help to identify possible processing errors within a specific batch of semen or on a particular day, should those errors occur. As sperm counting procedures become more refined, routine counting can be used to monitor subtle changes in daily semen processing that might affect the number of sperm packaged in a straw. Hemacytometers are widely used for routine sperm counts, but the equipment is slow, and multiple measurements of each sample are needed. Single hemacytometer counts are not highly accurate; because of inherent errors in the technique, Freund and Carol (13) found that mean differences of 20% were not uncommon between duplicate sperm count determinations by the same technician. Electronic counters provide much more rapid counting, are easier to use, and give more repeatable results among technicians. However, those instruments tend to include in the sperm count any somatic cells present, immature sperm forms, cytoplasmic droplets, debris, and bacteria, thereby inflating the concentration value (19). Currently, the primary method used by the AI industry to estimate sperm concentration is spectrophotometric determination of turbidity of a semen sample using an instrument previously calibrated for sperm concentration with a hemacytometer or Coulter counter (1). This approach is only as accurate as the methods used for spectrophotometer calibration. New, more accurate methods for sperm count determinations are being sought to replace the older ones. Some laboratories are trying the Maklerm counting chamber (Seif- Medical, Haifa, Israel) and other improved hemacytometers, such as the MicroCellTM (Fertility Technologies, Inc., Natick, MA); however, these techniques will likely have standard lems similar to those associated with the standard hemacytometers. It may be argued that when comparing fluorescent microscopy assays with flow cytometry, one is examining patterns of fluorescence rather than fluorescence intensity, i.e., the flow cytometer is not capable of discriminating sperm which have a fluorescent marker bound to the equatorial segment or over one of the acrosomal membranes (Parinaud et al., 1993; Mortimer and Camenzind, 1989; Mortimer et al., 1987). Tao et al. (1993) compared flow cytometry and epifluorescent microscopy with various lectins and indicated that there is no significant difference between the two methodologies for detection of the acrosome reaction. However, it has been argued that lectins do not bind specifically to the acrosomal region of the sperm (Purvis et al., 1990; Holden and Trounson, 1991) and that other binding sites can be easily distinguished by epifluorescence microscopy, whereas flow cytometry identifies the signal from the entire sperm. Additionally, conventional light microscopic semen assessment is increasingly being replaced by fluorescent staining techniques, computer-assisted sperm analysis (CASA) systems, and flow cytometry (PenËÅ"a et al., 2001; Verstegen et al., 2002). Additional advantages over existing techniques are that this approach is faster than the hemacytometer and that cellular debris, fat droplets, and other particulate material in extended semen are not erroneously counted as sperm, as often occurs with electronic cell counters. This method can also be used to determine the number of somatic cells in a semen sample. Viability The viability of spermatozoa is a key determinant of sperm quality and prerequisite for successful fertilization. Viability of spermatozoa can be assessed by numerous methods, but many are slow and poorly repeatable and subjectively assess only 100 to 200 spermatozoa per ejaculate. Merkies et al. (2000) compared different methods of viability evaluation. They concluded that Eosin-nigrosin overestimate viability while fluorescent microscope and flow cytometry estimate similar trend of viability. Currently flow cytometric procedures have been developed which simultaneously evaluate sperm cell viability, acrosomal integrity and mitochondrial function. This method has been successfully used for assessing spermatozoa viability in men (Garner and Johnson, 1995), bulls (Garner et al., 1994; Thomas et al., 1998), boars (Rodrà ­guez-Martà ­nez, 2007; Garner and Johnson, 1995; Garner et al., 1996), rams (Garner and Johnson, 1995), rabbits (Garner and Johnson, 1995), mice (Garner and Johnson, 1995; Songsasen et al., 1997), poultry and wildfowl (Donoghue et al., 1995; Blanco et al., 2000) and honey bees (Collins and Donoghue, 1999; Collins, 2000) and in fish (Martin Flajshans et al., 2004). Considerable information has accumulated on the use of fluorescent staining protocols for assessing sperm viability (Evenson et al., 1982). The SYBR 14 staining of nucleic acids, especially in the sperm head, was very bright in living sperm. Good agreement was observed between the fluorescent staining method and the standard eosin-nigrosine viability test; the flow cytometric method showed a precision level higher than that of the manual method. One of the first attempts to assess sperm viability utilized rhodamine 123 (R123) to assess mitochondrial membrane potential and ethidium bromide to determine membrane integrity using flow cytometry (Garner et al., 1986). Other combinations that have been used to examine the functional capacity of sperm are carboxyfluorescein diacetate (CFDA) and propidium iodide (PI) (Garner et al., 1988; Watson et al., 1992); carboxydimethylfluorescein diacetate (CMFDA), R123, and PI (Ericsson et al., 1993; Thomas and Garner, 1994); and PI, pisum sativum agglutinin (PSA), and R123 (Graham et al., 1990). At present, one of the most commonly used viability stain combinations is SYBR-14 and PI, sold commercially as LIVE/DEAD Sperm Viability kit (Molecular Probes Inc., OR, USA). When used in combination, the nuclei of living sperm fluoresce green (SYBR-14) and cells that have lost their membrane integrity stain red (PI). This staining technique has been used in a number of species, including the boar (Garner and Johnson, 1995; Saravia et al.,2005, 2007,2009). Although species differences do exist in the function of spermatozoa, the Live/Dead stain may similarly have no adverse affect on fertilization in the equine, although it remains to be tested in this species. Recently a new instrument (Nucelocounter-SP100) has been used to evaluate boar sperm concentration [11]. Due to its compact size and its relatively inexpensive purchase price, this instrument could be useful for field measurements of both concentration and viability. This instrument was considered to be a useful instrument for rapidly measuring stallion sperm concentration and viability (Morrell et al., 2010). Fluorescent probes such as H33258, requiring flow cytometric analysis with a laser that operates in the ultraviolet light range, are less commonly used as this is not a standard feature on the smaller analytical machines. However, one alternative is to use a fluorometer. A fluorometer is a relatively low-cost piece of portable equipment that permits a rapid analysis to be carried out on a sample. Januskauskas et al. (2001) used H33258 to detect nonviable bull spermatozoa by fluorometry and found a negative correlation between the percentage of damaged cells and field fertility. Another option is fluorescent attachments for computer-assisted semen analysis devices. For example, the IDENT fluorescence feature of the Hamilton-Thorne IVOS permits staining with H33258 allowing an assessment of sperm viability to be made along with motility. Fluorochromes used to assess sperm viability by either approach can be used in combination with each other. For example, when CFDA is used along with PI, three populations of cells can be identified: live, which are green; dead, which are red; and a third population which is stained with both and represents dying spermatozoa. Almlid and Johnson (1988) found this combination useful for monitoring membrane damage in frozen-thawed boar spermatozoa during evaluation of various freezing protocols. Harrison and Vickers (1990) also used this combination with a fluorescent microscope and found it to be an effective indicator of the viability of fresh, incubated or cold-shocked boar and ram spermatozoa. Garner et al. (1986) used this combination to stain spermatozoa from a number of species, but at that time could not find a relationship between bull sperm viability detected by CFDA/PI and fertility. Flow cytometry for assessment of sperm viability appears to be a valuable tool for the AI industry. When a high number of sperm is packed in each insemination dose, the effect of selecting the best ejaculates according to sperm viability has a relatively limited effect on NRR56. However, sperm viability might be more important when combined with low-dose inseminations. The FACSCount AF flow cytometer also determines sperm concentration accurately and precisely during the same analysis (Christensen et al., 2004a). The combination of assessment of sperm viability and concentration appears to be useful in the improvement of quality control at AI stations. Because of the results of this trial, this method has been implemented by Danish AI stations (Christensen et al., 2005). Relatively bright fluorescence was found also in the mitochondrial sheath of living sperm. The mechanism by which SYBR-14 binds to the DNA is not known. It is know that PI stains nucleic acids by intercalating betwee n the base pairs (Krishan, 1975). Viability stains have also been used in association with fluorescently labeled plant lectins to simultaneously assess the plasma membrane integrity and the acrosome integrity (Nagy et al., 2003). Assessment of viability using SYBR-14 dye does not damage spermatozoa, since Garner et al. (5) demonstrated that insemination of boar spermatozoa stained with SYBR-14 into sows did not compromise fertilization or the development of flushed porcine embryos in culture. Non-viable cells can be determined using membrane-impermeable nucleic acid stains which positively identify dead spermatozoa by penetrating cells with damaged membranes. An intact plasma membrane will prevent these products from entering the spermatozoa and staining the nucleus. Commonly used examples include phenanthridines, for example propidium iodide (PI; (Matyus, 1984) ethidium homodimer-1 (EthD-1; (Althouse et al., 1995), the cyanine Yo-Pro (Kavak, 2003) and the bizbenzimidazole Hoechst 33258 (Gundersen and Shapiro, 1984). Wilhelm et al. (1996) compared the fertility of cryopreserved stallion spermatozoa with a number of laboratory assessments of semen quality and found that viability, as assessed by flow cytometry using PI, was the single laboratory assay that correlated with stallion fertility. Changes in sperm membrane permeability Detection of increased membrane permeability is employed in different cell types to distinguish different status of membrane organization (Cohen, 1993; Ormerod et al., 1993; Castaneda and Kinne, 2000; Reber et al., 2002). Sperm plasma membrane status is of utmost importance due to its role, not only as a cell boundary, but also for its need for cell-to-cell interactions, e.g. between spermatozoa and the epithelium of the female genital tract and between the spermatozoon and the oocyte and its vestments (for review, see Rodriguez-Martinez, 2001). Membrane integrity and the stability of its semipermeable features are prerequisites for the viability of the spermatozoon (Rodriguez-Martinez, 2006). However, cryopreservation, whose purpose is to warrant sperm survival, causes irreversible damage to the plasma membrane leading to cell death in a large number of spermatozoa (Holt, 2000) or, in the surviving spermatozoa, to changes similar to those seen during sperm capacitation, thus shorten ing their lifetime (Perez et al., 1996; Cormier et al., 1997; Maxwell and Johnson, 1997; Green and Watson, 2000; Schembri et al., 2000; Watson, 2000). During the freezing process, cells shrink again when cooling rates are slow enough to prevent intracellular ice formation as growing extracellular ice concentrates the solutes in the diminishing volume of non-frozen water, causing intracellular water exosmosis. Though warming and thawing, the cells return to their normal volume. Thus, it is important to know the permeability coefficient of the cells to cryoprotectants, as well as the effect of cryoprotective agents on the membrane hydraulic conductivity. Classical combination of probes allows discrimination of two or three subpopulations of spermatozoa, i.e. live, dead and damaged depending on the degree of membrane integrity (Eriksson RodrÄ ±Ã‚ ´guez-MartÄ ±Ã‚ ´nez, 2000). A new, simple and repeatable method to detect membrane changes in all spermatozoa present in a boar semen sample, by use of markers (combination of SNARF-1, YO-PRO-1 and ethidium homodimer) used to track changes in sperm membrane permeability, has been developed recently by our group (Pena et al., 2005). In determined physiological or pathological situations, live cells are unable to exclude YO-PRO-1, but are still not permeable to other dead-cell discriminatory dyes, like propidium iodide or ethidium homodimer. YO-PRO-1 is an impermeable membrane probe and can leak in, only after destabilization of the membrane, under conditions where ethidium homodimer does not. Because several ATP-dependent channels have been detected in spermatozoa (Acevedo et al. , 2006), it seems plausible that this is a result of the silencing of a multidrug transporter. This m