PCR and Southern Blotting: Applications in Medicine

PCR and Southern Blotting: Applications in Medicine Introduction The 1970s introduced an inspirational technique in which a specific sequence could be determined from a sample of DNA via the process of Southern blotting. This method, named after Edwin Southern, provided the basis for a range of common laboratory techniques such as western blotting, eastern blotting and northern blotting [1]. Shortly after, in 1983, a revolutionary method was invented by Kary Mullis, called the polymerase chain reaction (PCR) [2]. This procedure was originally used to amplify and identify DNA sequences in the human genome. Its use in genetic analysis was immediately recognised, as one of the first publications of its use was of prenatal diagnosis of sickle-cell anaemia [3]. Since then it has been manipulated and a whole array of techniques have been derived from this invention. Both PCR and Southern blotting have been used widely in understanding and identifying microbes which in turn assists the diagnosis and management of patients suffering from infectious diseases. PCR Procedure The sample of DNA is heated up to 90Â °C to separate the two strands of DNA thereby exposing the nucleotide bases on each strand. A primer is then annealed to each strand from the 5 region at around 60Â °C and the temperature is increased once more. The thermostable DNA polymerase enzyme, Taq, prevents contamination by binding free complementary base pairs to the original strand of DNA at temperatures up to 70Â °C. The strands are then cooled and double the amount of DNA is synthesised, and the cycle restarts until a sufficient amount of DNA is produced. Reverse-transcriptase PCR (RT-PCR) is used when the original sample of RNA is transcribed so that DNA is the product of amplification. The sensitivity of PCR is great, as detection is from a single nucleotide base whilst its quantitative ability is derived from the proportional expansion of amplified DNA from its original size [5]. Southern Blot Procedure Southern blotting begins with a sample of DNA which is first broken up by a restriction endonuclease into smaller, varying fragments. The DNA is then placed into wells to undergo agarose gel electrophoresis where the fragments diffuse across a polarised field according to their size. The DNA is denatured by sodium hydroxide and transferred to a sheet of nitrocellulose or nylon and incubated with a hybridisation probe of single-stranded DNA. This radiolabelled probe binds to the exposed complementary base pairs and can be detected by autoradiography [6]. Southern Blotting Applications in Medical Microbiology Southern blotting is primarily used for DNA fingerprinting, gene sequencing and genetic engineering. It has been used in the identification of strains in microbes such as concluding the type of human papillomavirus extracted from a condyloma. However in this case it provided to be unreliable as it produced false-negatives, as PCR and in-situ hybridisation were deemed to be more efficient [7]. Another use of Southern blotting was in the detection of a strain of Listeria monocytogenes. In this study it was deemed an important technique in verifying species identification and in the characterisation of epidemic strains [8]. This method can be used to DNA fingerprint most microbes and determine a diagnosis and treatment for a patient suffering from their pathogenicity. However it is found to be too laborious, time consuming and requires large amounts of high quality DNA for most routine laboratories yet variations of this technique are still widely performed. The western blot, which uses antibodies as the probe to detect proteins instead of DNA, is a confirmatory test in the diagnosis of a human immune-deficiency virus(HIV) infection [9]. Since the established use of PCR there has become a decreased need for these laborious techniques in DNA sequencing [10]. PCR Applications in Medical Microbiology PCR can be used in detecting the genetic sequence of all microbes. It is useful in detecting organisms in early cultures where organisms are otherwise difficult to isolate, for example in enteroviruses RT-PCR is more sensitive than culture and the gold standard is detection of the this genome in cerebral spinal fluid (CSF) by PCR [11]. PCR is also used in detecting genes encoding antibiotic resistance such as in Helicobacter pylori and Methicillin-resistant Staphylococcus aureus. However its use is currently unsuitable for the diagnosis of H. pylori as clinical samples may contain inhibitors which can generate false-negatives10. PCR is used in quantifying the viral load of HIV within an infected person therefore determining the effectiveness of their treatment. The problem occurs when the genome sequence of the HIV changes therefore the PCR method needs to be altered and the current test would prove useless9. Currently immunoassays are used in the diagnosis of a HIV infection, however early in infection there is a variable period of time until anti-HIV antibodies can be detected and this provides a potential area of identification of the HIV by PCR [12]. Pitfalls of PCR include that the sample must be kept cold during storage and transport to the laboratory, the expertise required for analysing and interpreting results as well as standardising between different laboratories and finally the expensive cost of the procedure compared to available techniques9. Conclusion The applications for PCR and Southern blotting are enormous however they have limitations which prevent it from being routinely used in the diagnostic laboratory. With the advance of technology both are being developed, especially PCR, and have endless applications in medical microbiology.

Employment Laws and HRM Strategy Essay

Describe the scenario that you have selected and its corresponding employment laws. The human resource scenario the writer selected is the need for seasonal employees who may be from another country. Employment laws for the state of Mississippi will be a base used to solve this issue. Mississippi Wage and Hour Laws from the Employment Law Handbook insures that seasonal workers are provided with a minimum wage, meals and breaks, vacation, sick leave, severance and holidays. Mississippi currently does not have any state laws requiring or regulating these requests. Mississippi uses federal guidelines as a base for establishing employment laws. The federal law as of today requires a minimum wage of 7.25 dollars. The federal law for overtime is also followed by Mississippi. Meals and breaks according to federal law are not defined. Breaks taken by an employee that are less than twenty minutes must be paid and lunch breaks thirty minutes or more do not need to be paid as long as an employee can do whatever he or she likes during this period according to the Department of Labor. Vacation leave is determined to the employer since Mississippi legislature does not have any established guidelines on this matter. Sick leave, Severance pay and holidays are all determined by an employer in the state of Mississippi due to established laws in this state (Mississippi Wage and Hour Laws, 2012). In August of 2012 Mississippi held a committee meeting on immigration reform. Governor Phil Bryant wanted to reaffirm a law preventing undocumented immigrants from recovering public benefits. The Mississippi Senate will have to take into consideration the United States Supreme Court rulings and federal appeals when drafting Mississippi legislature for immigration laws. The Mississippi state immigration law would have to comply with E-Verify laws and constitutional laws are not violated. President Barack Obama passed a deportation deferral to immigrants earlier this year but states can pass their own immigration reform laws such as Arizona and Alabama (Cherry, 2012). The United States Department of Labor established an act called The Migrant and Seasonal Agricultural Worker Protection Act or (MSPA) in order to protect these types of workers. The act protects employment related migrant and seasonal workers. The MSPA ensures that each nonexempt farm contractor and agricultural seasonal association disclose their terms and conditions of employment to each migrant worker. The terms and conditions of employment must be explained to the migrant and seasonal worker at the time of recruitment (Department of Labor, 2012). The job must description must also explain worker protection, amount of wages due to a migrant worker and provide an itemized statement of earnings and deductions (Department of Labor, 2012). The Migrant and Seasonal Agricultural Worker Protection Act also guide housing and transportation. The federal guide line ensures that housing and transportation provided meets federal and state guidelines with proper licensure and insurance which is enforced by the wage and hour division. The federal guidelines require records must be kept for at least three years which includes name, permanent addresses and social security numbers (Department of Labor, 2012). The Fair Labor Standards Act or (FLSA) requires that workers on large farms are paid minimum wage but they are excluded from overtime premium pay. The FLSA prevents children under the age of sixteen from working during school hours and working certain jobs deemed too dangerous. Children that are employed on their families’ farm are exempt from the Fair Labor Standards Act (Department of Labor, 2012). The Immigration and Nationality Act (INA) require employers that want to use foreign temporary workers with H-2A visas get a labor certificate from the Employment and Training Administration. The certificate ensures that there are not enough able, willing and qualified United State workers available to do the work. The Immigration and Nationality Act standard of the H-2A program is enforced by the wage and hour division (Department of Labor, 2012). Recommend a plan to manage the HRM situation within the confines of the law. Justify your approach to your scenario to HR management. The human resource management plan the writer recommends would be based on performance. Job performance and task performance represents what an employee contributes to an organization that employs them. Migrant workers would benefit from this type of grading system if the work performance is factory based. Task performance is the product of employees performing actions leading to the production of raw materials into goods and services. Studies have been performed to show job performance management provides feedback and measurement improves employee performance leading to better organizational performance (Stewart, Brown, 2011). The writer believes migrant workers may show citizenship behavior. Employees often show citizenship behavior by going the extra mile for the company, volunteering for special projects, following organizational procedures even if they are not convenient for the employee and defending the business where the employee works (Stewart, Brown, 2012). The writer would focus on counterproductive behavior before it starts. Counterproductive behavior is behavior from an employee that is harmful to a business. An employee can exhibit two types of counterproductive behavior: 1) production deviance 2) property deviance occurs when employee sabotage equipment, destroys materials and products and misuse expense accounts. The counterproductive behavior can also be extended to gossiping about coworkers and showing favoritism. The writer would create a manual of policies and procedures explaining how this behavior is prohibited and have disciplinary consequences set in place if this behavior is exhibited. The writer would not use performance appraisals as much. The main goal would be to give feedback to an employee and listen to any problems they may be having. State how a competitive advantage may be gained by ensuring HRM practices meet the necessary employment laws. A competitive advantage can be gained by ensuring human resource strategies are followed. A company must have an employee handbook, benefits package, job descriptions and a payroll system in place to ensure human resource strategies are at an advantage to the company. The steps above can be followed also to keep in compliance with legal employment issues. Laws and regulations are important to ensure equal rights to disabled worker, employment eligibility for migrant workers with the use of an I-9 form, drug screening and work place harassment. A competitive strategy can lead to effective human resource department and a successful business (Managing Employees, 2012). Speculate what might happen if the situation was not handled in accordance with the appropriate employment laws. A business that does not follow employment laws can face state and federal criminal charges. A business can avoid costly law suits by following laws to ensure employees are treated fairly. A company’s compliance can save money that would be used to repair a damaged reputation. Specific laws protect employees from immutable characteristics such as sex, race, age and religion. Characteristics such as tattoos, nose piercings and long hair are not covered by federal and state laws. State laws can set a basis by not discriminating against marital status or sexual orientation and may not conflict with federal laws. The writer speculates that if state and federal discrimination laws are broken a company can face many damaging hardships (Stewart, Brown, 2012). References Cherry, Daniel (2012). Lawmakers Gearing Up for Immigrant Reform Battle Retrieved from http:www.mpbonline.org on November 18, 2012 Stewart, G., Brown, K. (2011). Human resource management (2nd ed.). Hoboken, NJ: John Wiley and Sons Managing Employees (2012). Retrieved from http:www.mississippi.org on November 18, 2012 Mississippi Wage and Hour Laws (2012). Employment Law Handbook, Retrieved from http:www.employmenthandbook.com on November 18, 2012 The Migrant and Seasonal Agricultural Worker Protection, (2012) Department of Labor Retrieved from http:www.dol.gov on November 18, 2012

The Developments in Adolescence and Emerging Adulthood

Nothing is constant for everything in the world is impermanent. Everything in this world eventually undergoes some form of change. As the world changes, the people also go through essential changes. Children suddenly grow into teenagers and the teenagers become adults. People grow old and the cycle goes on. Thus, various disciplines, such as psychology, categorize human life into several stages, which are determined by age. Each stage has its distinguishable characteristics that aim to describe the attributes of the particular group of individuals.Every person passes through each stage of life. This includes infancy, childhood, adolescence, adulthood, and old age. People pass through every stage of development until they reach old age. Under each stage, they experience vital changes in the different aspects of their lives, including physical, cognitive, behavioral, and social aspects of development. Adolescence is considered one of the most crucial stages of the human life. It is the pivotal point where people search for their individuality and determine who they are.This is the reason why the term â€Å"teenage crisis† was developed, as adolescence is the period when a person goes through many significant changes physically, socially, and emotionally. Physical Development in Adolescence Adolescents include people under the age bracket of 13 to 18 years old. During this stage, people experience mixed emotions such as happiness and anxiety, as this the time when people outgrow their childish ways and start facing responsibilities. Puberty marks the biological and physiological maturation of one's body.Various changes could be seen in children who enter this transition. Growth is one of the changes a person experiences during the transition between childhood and adult life. In the case of girls, they begin to rapidly increase in height and achieve almost 98% of their full height at the age of 16. On the other hand, the boys’ period of rapid growth i s at the age of 13 until they reach 20. Many of the teenagers also tend to have a tall and lanky appearance. This is because the skeletal system outpaces the muscular system and the bones elongate while the muscles stay the same (Hatfield, 2007).Girls also go through a significant change during this stage. They start their menstrual cycle which signals readiness for pregnancy. Their breasts also start to develop, and they achieve adult appearance by the age of 16. Boys also achieve their adult appearance by the age of 16. Their muscle strength and coordination are well developed. Their voice also changes, which sometimes causes them embarrassment (Hatfield, 2007). Because of the sudden change in the body, there is also a change in the nutritional needs of adolescents.Since they are growing rapidly and engage in various activities, they need to have more energy and calories to complement in their hyped lifestyle. Teenagers also become aware of sexual changes and sexual maturity. They suddenly become curious in the aspect of sex and sexuality. Society's culture has a significant impact on the perception of adolescents toward sex. Sex is perceived differently in various cultures and this greatly influences adolescents’ developed view on sex. Sexual orientation is also developed during this stage.Adolescents begin to look for their self and establish their identity as they discover their taste and preference. There are various perspectives to describe the adolescence phase. For instance, parents see this stage as full of unsolicited outbursts and rebellious acts. Meanwhile, biological perspectives explain that the adolescence is a stage of turbulence that closely resembles the period when people evolved from savages to civilized beings. Cognitive Development in Adolescence During adolescence, people also experience vital developments in the cognitive aspect.Cognitive maturation includes a wide scope of newly found abilities and functions of the brain, such as the significant change from concrete to abstract thinking. This capability allows adolescents to draw logical conclusions in various situations (Sadock Kaplan, & Sadock, 2007). Jean Piaget also attempts to explain the changes in the cognitive aspect of an individual when he reaches adolescence. According to Piaget, adolescents enter the formal operational stage wherein their cognitive capacity develops, providing them with abstract and scientific thinking skills.Its difference with the concrete operational thinking in children is that children just see the plain reality. They are not concerned with the underlying problems. Adolescents tend to be more inquisitive and probing when it comes to dealing with problems. Statistical data show that about 40 to 60% of adults fail the Piaget's operational problems. This is because adults practice formal operational thinking in their field of expertise or in the field where they have extensive experience. Another reason is adolescents and ad ults do not always practice formal operational thinking.They tend to rely on their intuitive judgments and conclusions. Emotional and Social Development in Adolescence During adolescence, a person is bound to experience some psychological and emotional difficulties while passing through puberty and adolescence. Some of the adolescents develop difficulty in the transition, which includes fear and anxiety and sometimes cause deep depression. It also includes confusion, identity crisis, and anxiety. These things are a common experience during adolescence, and many adolescents do not know how to adapt and conform to the sudden changes.Nevertheless, there are some adolescents who still maintain vital and intimate relationships with others. Some of them are also easily adapt to the changes and develop a positive outlook towards their personal identity (McInerney, 2006). Erik Erikson was the first to discover that identity is one of the major premises that affect the individual's acquisiti on of healthy and productive adult life. The description of oneself, or identity, includes who a person is and what he or she values, his or her goals, and objectives in life. Erikson then developed a theory called identity versus identity confusion.The theory explains that fruitful outcomes during the early stages will render positive resolutions. Erickson also explained that adolescents may also experience identity crisis—a period of confusion and distress. Adolescents tend to explore alternatives before they draw a conclusion. Erikson also added that those who go through â€Å"soul searching† most often settle for a mature identity. There are various things that influence the development of identity. An individual’s identity is molded and shaped by these factors. Parents, peers, school and community are the factors that affect identity.Parental support and encouragement play significant roles in the formation of identity. Being welcomed to a group of peers bo osts people’s self-esteem and relatedness. Their school and community define the environment where they live which affects their attitude and behavior. Emerging Adulthood This period is not an extension of one's adolescence, nor is this the period of young adulthood. During this period, individuals experience less parental control and supervision and they engage in independent exploration. Young adulthood implies that a person is in the early stage of adulthood.Emerging adulthood is a transition after adolescence wherein one is too old to behave like a typical teenager but too young to assume an adult way of life and involve themselves sin adult issues such as marriage and rearing of children (Arnett, 2004). Emerging adulthood is a time when a lot of uncertainties and queries are supposed to be entertained in preparation for the real adult life. It is also considered as one of the most stressful times in one's life, which involve a period of constant exploration in education, jobs and partners. Emerging adults also devote their full attention on looking for alternative roles, values, and behavior.Thus, emerging adulthood is also a stage wherein many changes occur in one’s life. These changes include cognitive development beyond Piaget's formal operational stage. Emotional and social changes also occur during this period.Their attitudes and values widen, as they express interest in different kinds of new fields and accept ethnic and cultural diversity. Conclusion Adolescence is both an exciting and anxious stage in human life. During this stage, individuals experience abrupt changes and developments. They experience physical, cognitive, social, and emotional changes in forming and defining individuality and personal identity.Emerging adulthood is the transition stage between adolescence and young adulthood. In this stage, individuals enjoy the carelessness in decision making. They are not truly engage in adult life. They explore different things l ike education, jobs, and intimate relationships. They also experience significant changes in the cognitive, social, and emotional aspects. All people go through the stages of the human life cycle. Individuals tend to be anxious and scared when they step into an unknown phase of life. Hence, people should take everything as a significant experience in their search for their real personal identity.ReferencesArnett, J.J. (2004). Emerging Adulthood: The Winding Road from the Late Teens Through the Twenties. Oxford: Oxford UP. Hatfield, N.T. (2007). Broadribb's Introductory Pediatric Nursing. Philadelphia: Lippincott Williams & Wilkins. McInerney, D.M. (2006). Developmental Psychology for Teachers: An Applied Approach. Australia: Allen & Unwin. Sadock, B.J., Kaplan, H.I. & Sadock, V.A. (2007). Kaplan & Sadock's Synopsis of Psychiatry: Behavioral Sciences/Clinical Psychiatry. Philadelphia: Lippincott Williams & Wilkins.

Essay On Teen Depression - 1321 Words

Word count: 1108 Summary: Teen depression is a serious issue that plagues the nation. The 3rd leading cause of death for young adults and teens is suicide, which can often be the result of depression. Olivia Reardon is one such victim of teen depression. Beginning high school, Olivia was Captain of the lacross and field hockey team as well as an honors student. This all changed when Olivia turned 15. Her mother slowly began to notice the subtle signs of depression in her daughter. These included irritability, anger, and even rage. Olivia was in a dower mood often and had lost interest in things which normally excited her. She handled this by bottling up her emotions. She thought that this was her problem, and she didn’t need to bother her†¦show more content†¦Therapy is always the first line of defense. Depression is a physical illness that affects the mind just like appendicitis affects the body. The social stigma that surrounds mental health issues is a barrier that keeps patients from treat ment and is something we as a society have to address. If you have a firearm in your home and you are afraid your family member may be suffering from depression, secure it. It is of the utmost importance that the person afflicted does not have a means to harm themselves or others. If you wish to use holistic approaches to combat depression, they are best used to supplement medicine and therapy. Light therapy and vitamins are effective supplements. Response: I have a very close friend who has tried to take his own life after a battle with depression. One thing that I think is overlooked is the roles anxiety and internalization of emotion play in depression. Suicidal thoughts and actions are often induced by the desire to escape extreme anxiety by any means necessary. This is how my friend described it to me. He also said that he felt like a bother to everyone and that his perpetual existence was only a burden on his family. These thoughts combined into a perfect storm. Luckily, he reached out to his parents and was hospitalized for several days. Many of our close friends and I were shocked to find out about this turn of events. We had no idea he was going through this deep, dark depression. The amount of friends and familyShow MoreRelatedEssay on Teen Depression1653 Words   |  7 PagesClinical depression is capable of ruining the future. In a few years, teenagers will be moving up in the world and beginning their lives as contributin g members of society. Each one of them will have a job, and will encounter countless other individuals. The dilemma that the world faces is about 17% of teenagers will suffer from depression before they become adults (Canada). This impacts connections not only in the political/business world, but their personal lives and growth as well. 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