Increasing Student Achievement through Preventing School...

Increasing Student Achievement through Preventing School Violence Introduction School violence is any type of violence, whether it is a simple threat on another student or a school shooting, that happens in the school environment. Student achievement is the efforts made by the student to work hard in the classroom. The purpose of this research paper is to examine how school violence impacts student achievement. It would seem that school violence would have a negative impact on student achievement. If this is the case then we need to do what we can to prevent violence in schools. Technology can be used to help control school violence. Information on School Violence Violence in our schools has become an increasing†¦show more content†¦It was found that males tended to fall victim to crime more so than females (Furlong Chung, 1995). Multi-victim students of violence were more likely to receive D’s and F’s whereas non-victim students tended to receive more A’s. Students have not experienced crime perceived their schools to be a safe environment and therefore were able to be more focused to their schoolwork than worrying about violence. Since they are able to focus better in school, their achievement level is considerably higher than a student who has experienced many incidents of violence. Multi-victim students of violence tend to be more worried at school and do not feel safe in their school environment. These insecure feelings leave the students paranoid and therefore are not able to focus on their schoolwork as well as non-victim student. In order for a student to be successful we need to eliminate all distractions from the school environment. Violence is a distraction and even though it can not be eliminated completely, we need to see to it that it we remove as much of it as we can from our schools. All students deserve the right to a safe school and to have an equal chance to be successful in life. 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Are You Really My Hero - 1216 Words

AJ Wells Professor Carly Smith ENGL 112 80N-N1-201420 13 December 2014 Are You Really My Hero? â€Å"Heroes have their scars. Some you can see, some you read about later on.† George Foreman Growing up we all had our dreams of winning the gold medal in an Olympic event, scoring that game winning basket, being a prize fighter, or hitting the homerun with bases loaded in a World Series. We imagined the feeling of success and glory but we also realized that few do accomplish such. Seeing such deeds are what creates our desires, determination and goals. The athletes that achieve such give us our heroes to aspire to attain such levels. We want the future generation of athletes to have these same dreams, goals and hopes of success that we grew up with. But are our current athletes the type of heroes we want our children and young athletes to look up to? Performance Enhancing Drugs are becoming the norm for many of our athletic heroes and modern day headlines are expounding on the wide spread usage among our elite athletes. Some authors such as Michael Lavin in â€Å"Sports and Drugs: Are the Current Bans Justified?† states that the expectations and demands place d upon today’s athletes are much greater than in the past and this pressure leads to using enhancements. Professional athletes are now paid large salaries so that more expectations are made of their abilities than may be achievable with natural workouts and practice of the past. There is pressure to stay in the gameShow MoreRelatedCharacteristics Of A Hero891 Words   |  4 Pages Tree traits of a hero â€Å"A hero is an ordinary individual who finds the strength to persevere and endure in spite of overwhelming obstacles.† - Christopher Reeve. Having heros in this world is really important. If we didnt have everyday heros our world would be a lot different. 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Counseling Ethics Samples for Students †MyAssignmenthelp.com

Question: Discuss about the Counseling Ethics Case of Susan Lim. Answer: Susan Lim is a prominent surgeon in Singapore who has performed several complex surgeries throughout her career. Due to her exceptional capabilities, she is famous all across the globe. She was head surgeon in the first liver transplant in Singapore, became the youngest person and first Singaporean to become the fellow of Trinity College, Melbourne. In 2001, she undertook the case of Pengiran Anak Hajah Damit, the sister of Brunei queen. In 2007, the patient died a d Dr. Susan Lim was accused of overcharging the patient with a very high charge. According to the media reports, Dr. Susan Lim was accused of making a bill of 40 million dollars during the period of 2001 to August 2007 (Asia One Health, 2016). In the later investigations, it was found that the bills constructed were astonishingly high than the normal circumstances. In 2013, doctor was prosecuted and found guilty. Singapore National Council filed a case against the Susan Lim and won the case eventually. The doctor was found guilty. During the judgment, the doctor presented her side by stating that she charged the patients in accordance to the services provided to them. She stated that although the prices were very high, but they were in accordance to the services. In order to excellent services to her client, she received training from all across the globe. In several instances, she herself paid for the services provided to her client such as sending the patient from Singapore to Brunei; therefore, she was only asking them from the client (John Harding, 2011). Moreover, her professional conduct can be scrutinized from the fact that in order to halt the legal processioning, she threatened the Singapore Medical Council that she can bring into light several unnecessary and embarrassing facts which can bring shame to the government and the royal family of Singapore. She stated that she knows the inconsistent relationship between Ministry of Health, Government of Brunei and the royal family of Brunei. The threat brought her a second disciplinary action. Identification of Ethical Dilemma The code of medical ethics regulation state that the doctors should not overcharge their patients. According to the professional ethics, doctors should charge their patient according to the services provided by them. However, same is not the case of Doctor Susan Lim. Moreover, overcharging is against the professional practices, as it refers to deceit and abuse of ones position. It can be critiqued that there was another moral issue wherein that the doctor tried to threaten the government to drop the accusations. Major Stakeholders of the Case: Upon the review of the ethical issue, the major stakeholders can be identifies as Brunei Royal Family, Singapore Medical Council, Singapore government and the doctor herself. Susan Lim Susan Lim was the primary stakeholder in the ethical issue. She was the performing doctor who took case of the patient for six years. She was of the viewpoint that she provided exceptional services to the patients, invited the foreign professionals and received training which made it acceptable for her to overly charge her patients (Youtube, 2011). Singapore Medical Council (SMC) Singapore Medical Council was also a major stakeholder in the case, as it is the responsibility of the organization to maintain the professional integrity among the healthcare professionals and doctors in the Singapore. Any breach of duty is against is breach of guidelines of the Singapore Medical Council. In the present situation, Dr. Susan Lim breached the guidelines of Singapore Medical Council and the organization should try to impose justified penalty for the unethical behavior (Broad, 2000). Brunei Family Brunei family is the victim in the present situation. The doctor exploited the family due to their excessive wealth and influence. The doctor tried to obtain as much fees as she possibly could as she knew that her client was capable of paying higher fees. When she faced trial, Susan Lim also tried to exploit the vulnerable relations between Brunei government and Royal family as she was aware of their relations (Timmons, 2012). Singapore Government It is the responsibility of the government to protect the society from any type of criminal or civil offence. The professional medical society of Singapore has made code of conduct for the healthcare professionals so that they work ethically. In the present case, the Singapore government is a stakeholder as it is its responsibility to maintain law and order in the country. Ethical Dilemma The ethical dilemma can be defined as a situation in which a person is conflicted between different courses of actions in which taking one action will result in transgressing some other moral imperative. In the present situation, the ethical dilemma of overcharging the patient has been identified. The doctor calculated the fees according to her perspective and presumed that the exceptional services provided to the client justify the fees imposed upon them. She singularly treated one patient and kept all her focus to the sister of the royal queen of Brunei (Garber, (2008). Therefore, in her perspective, the fees imposed upon the client were justified. However, the royal family of Brunei was displeased with the doctors fees as they were exceptionally high. They presumed that the doctor was charging them with high fees die to their wealth. Moral Theories In the recent years, a large number of moral theories have emerged to examine whether certain actions are morally fit or not. These theories try to provide a systematic method to evaluate whether certain actions are morally right or wrong. However, different moral theories are dependent upon different perspectives and several times contradict each other. In the present section, the ethical dilemma of Doctor Susan Lim is examined under different ethical frameworks. Utilitarianism Utilitarianism is the most popular and common ethical theory which states that the all those actions are ethical which have better outcomes for the society. The utilitarianism is a version of consequentialism and states that only the result of certain actions can be categorized as right or wrong. In this framework, the interest of all the stakeholders is considered equally. There are certain benefits and disadvantages of utilitarianism (Dreier, 2009). The advantages of utilitarianism are that it promotes equality and happiness of all the people in the world and assist the people in taking tough decisions. In most of the complex and tough decisions, the emotions and the desire of the people become predominant; however, adopting utilitarianism approach assist the people in thinking rationally and taking the right decision (Jacobs, 2008). When this theory is applied to the case of Doctor Susan Kim, it can be evaluated that the action is morally justified. The overcharging of fees was ethically wrong as it was only beneficial for the doctor. The client was wronged as the fees were astonishingly high and determined according to his affluence. Moreover, the action will be a bad influence on the society as it is a breach of the moral code of conduct of the healthcare professionals. Teleology The teleology theory of morality states that the morality of the actions depends on the expected outcomes of the actions. Teleology theory states that if a person is following a certain course of action to achieve good from the situation, then that action is justified (Hinman, 2012). The teleology theory has certain disadvantages like it allows immoral actions on the pretext of better consequences. The consequences of most of the actions are quite uncertain; however, it allows the people to take morally unjustified actions (Alexandra Miller, 2009). According to this theory, the actions of Susan Lim are morally wrong as she was not focused on the treatment of the patient rather than increasing the probability of charging her client in the maximum possible manner. Although she provided exceptional services to the patients, pricing her client highly was always in her mind. Counseling Process Doctor Susan Lim should be provided counseling on the moral justifications of her action. Adopting a morality framework and examining the actions through it can assist the doctor in taking morally right actions. Decision-Making Process In the daily life of professionals, it is important that the people should take morally correct and ethical decisions. In order to take morally correct and right decisions, the professionals need a decision making framework. In the present section, an eight-step decision making model is proposed which can assist doctor in ethical decisions in the near future. According to this model, the decision making process has eight phases. Phase 1: In this phase, the problem is defined. In good decision-making, the problem definition is very important. It assist person in establishing clarity on the actual problem and examining various viewpoint of different stakeholders. In this phase, the root or the underlying causes of certain problems is identified. In addition to it, in this phase the limiting assumptions, system and the interface boundaries and the stakeholder issues are identified (McKee, Kemp Spenxce, 2012). Phase 2: In the second phase of the model, the requirements for the acceptable solution are identified. The Acceptable solution must meet certain requirements and address the underlying problem. The requirements define what the solution must be capable of doing. Phase 3: In the third phase, goals and objectives are defined which can be achieved by the solution. It refers to the broad statements of intent and value with the decision. Phase 4: In the fourth phase, the person should ponder on different alternatives which can be used to address the problem statement. In good decision making, it is important to evaluate different alternatives according to their effective outcomes. In most of the cases, the alternatives are evaluated according to their ability to meet the requirements and goals of the organization. Phase 5: In the fifth phase, different criterions are defined according to which the alternatives to the solution are evaluated. It is important to define discriminating criterion which determines how well each goal is able to achieve the result. Phase 6: In the sixth phase, a decision making tool must be developed which is able to select the best route for the solution. The decision-making tool should be based on the complexity of the problem (Mowen, Hansen Heitger, 2016). Phase 7: In this phase, the selected alternative is implemented to achieve the solution. The person should also monitor the solution for its results. Phase 8: In the last phase, the implemented solution is measured for its performance. After the evaluation, the person can make amendments in the decision so that the desired outcomes can be achieved (Mowen, Hansen Heitger, 2016). Conclusion It can be concluded that the course of action taken by Doctor Susan Lim is unethical and immoral. It is due to the fact the doctor overcharged her patient excessively. The moral dilemma of Susan Lim has been evaluated according in utilitarianism and the teleological moral framework. According to the utilitiarism theory, all the actions are justified which have better outcomes for the society. In addition to it, the teleological moral theory states that if the intentions of the people are good then the actions are justified. The action can be categorized as immoral according to both theories. Upon the evaluation of situation, it has been determined that the action was immoral. The doctor can be provided counseling with the help of decision making framework. The eight step decision making framework has been suggested will be beneficial in the future decision making process of the organization. References Asia One Health. 2016. Surgeon billed Brunei patient $40m over 4 years. Retrieved December 10, 2016 https://health.asiaone.com/health/health-news/surgeon-billed-brunei-patient-40m-over-4-years Broad, C.D. (2000). Five Types of Ethical Theory. Psychology Press.Chew, R. (2011). DOCTORS FEES AFTER SUSAN LIMS CASE Implications for the Medical Profession. Retrieved December 10, 2016 https://www.sma.org.sg/UploadedImg/files/Publications%20-%20SMA%20News/4511/Insight.pdf John Harding. (2011). Dr Susan Lims threatening letter to Foreign Minister George Yeo. Retrieved December 10, 2016 https://johnharding.com/2011/03/dr-susan-lims-threatening-letter-to-foreign-minister-george-yeo/ Youtube. (2011). Singapore Top surgeon 'threatened' MFA - 28Mar2011. Retrieved December 10, 2016 https://www.youtube.com/watch?v=xWuWLuSZ95Y Garber, P.R. (2008). The Ethical Dilemma. Human Resource Development. Timmons, M. (2012). Moral Theory: An Introduction. Rowman Littlefield Publishers. Jacobs, J. (2008). Dimensions of Moral Theory: An Introduction to Metaethics and Moral Psychology. John Wiley Sons. Alexandra, A., Miller, S. (2009). Ethics in Practice: Moral Theory and the Professions. UNSW Press. McKee, A., Kemp, T., Spenxce, G. (2012). Management: A Focus on Leaders. Pearson Higher Education AU. Mowen, M.M., Hansen, D.R., Heitger, D.L. (2016). Managerial Accounting: The Cornerstone of Business Decision-Making. Cengage Learning. Dreier, J. (2009). Contemporary Debates in Moral Theory. John Wiley Sons. Hinman, L.M. (2012). Ethics: A Pluralistic Approach to Moral Theory. Cengage Learning.

Real Time Pcr free essay sample

PROBE-BASED DETECTION SYSTEMS14 Hybridization probes (also called FRET probes)16 MELTING CURVE ANALYSIS16 Multiplex real-time PCR18 APPLICATIONS OF REAL TIME PCR18 GENE EXPRESSION ANALYSIS18 SNP GENOTYPING19 HIV DETECTION19 CYSTIC FIBROSIS (CF) DETECTION:20 THE ADVANTAGES OF REAL-TIME PCR20 THE DISADVANTAGES21 REFRENCES21 REAL TIME PCR TRADITIONAL PCR The polymerase chain reaction (PCR) is one of the most powerful technologies in molecular biology. Using PCR, specific sequences within a DNA or cDNA template can be copied, or â€Å"amplified†, many thousand- to a millionfold. In traditional (endpoint) PCR, detection and quantitation of the amplified sequence are performed at the end of the reaction after the last PCR cycle, and involve post-PCR analysis such as gel electrophoresis and image analysis. REAL-TIME QUANTITATIVE PCR (qPCR) In real-time quantitative PCR (qPCR), the amount of PCR product is measured at each cycle. This ability to monitor the reaction during its exponential phase enables users to determine the initial amount of target with great precision. WHAT’S WRONG WITH AGAROSE GELS? * Poor precision. * Low sensitivity. Short dynamic range lt; 2 logs. * Low resolution. * Non-automated. * Size-based discrimination only * Ethidium bromide staining is not very quantitative REAL TIME PCR VS PCR . BASIC PRINCIPLE Quantitative PCR  is carried out in a  thermal cycler  with the capacity to illuminate each sample with a beam of light of a specified wavelength and detect the fluorescence emitted by the excited  fluorochrome. The thermal cycler is also able to rapidly heat and chill samples thereby taking advantage of the physicochemical properties of the  nucleic acids  and  DNA polymerase. The PCR process generally consists of a series of temperature changes that are repeated 25 – 40 times, these cycles normally consist of three stages: the first, at around 95  Ã‚ °C, allows the separation of the nucleic acid’s double chain; the second, at a temperature of around 50-60  Ã‚ °C, allows the alignment of the primers with the DNA template;  the third at between 68 72  Ã‚ °C, facilitates the  polymerization  carried out by the DNA polymerase In real-time PCR, * the amount of DNA is measured after each cycle by the use of fluorescent markers that are incorporated into the PCR product. The increase in fluorescent signal is directly proportional to the number of PCR product molecules (amplicons) generated in the exponential phase of the reaction. * Fluorescent reporters used include double-stranded DNA (dsDNA)- binding dyes, or dye molecules attached to PCR primers or probes that are incorporated into the product during amplification. * The change in fluorescence over the course of the reaction is measured by an instrument that combines thermal cycling with scanning capability. By plotting fluorescence against the cycle number, the real-time PCR instrument generates an amplification plot that represents the accumulation of product over the duration of the entire PCR reaction (Figure 1). Figure 1—Amplification plots are created when the fluorescent signal from each sample is plotted against cycle number; therefore, amplification plots represent the accumulation of product over the duration of the real-time PCR experiment. The samples being amplified in this example are a dilution series of the template. TYPES OF PCR Quantitative PCR| Qualitative qPCR| A specific or non-specific detection chemistry allows the quantification ofthe amplified product. | In qualitative qPCR, the goal is to detect the presence or absence of a certain sequence. | The amount detected at a certain point of the run is directly related to theinitial amount of target in the sample| For virus sub-typing and bacterial species identification. Can also be used for allelic discrimination between wild type and mutant, between different SNPs or between different splicing forms. | common pplications of quantitative PCR are gene expression analysis, pathogen detection/quantification and microRNA quantification| Different fluorophores can be used for the two alleles, and the ratio of the fluorophores signals correlates to the related amount of one form compared to the other one. | Quantitative PCR software uses the exponential phase of PCR for quantification. | Specific detection methods such as Double-Dye probe systems are more ofte n used for theseApplications| Overview of real-time PCR Real-time PCR is a variation of the standard PCR technique used to quantify DNA or RNA in a sample. Using sequence-specific primers, the relative number of copies of a particular DNA or RNA sequence can be determined.. Quantification of amplified product is obtained using fluorescent probes or fluorescent DNA binding dyes and real time PCR instruments that measure fluorescence while performing temperature changes needed for the PCR cycles. qPCR STEPS There are three major steps that make up a qPCR reaction. Reactions are generally run for 40 cycles. 1. Denaturation—The temperature should be appropriate to the polymerase chosen (usually 95 °C). The denaturation time can be increased if template GC content is high. 2. Annealing—Use appropriate temperatures based on the calculated melting temperature (Tm) of the primers (5 °C below the Tm of the primer). 3. Extension—At 70–72 °C, the activity of the DNA polymerase is optimal, and primer extension occurs at rates of up to 100 bases per second. When an amplicon in qPCR is small, this step is often combined with the annealing step using 60 °C as the temperature. BASICS OF REAL TIME PCR Baseline – The baseline phase contains all the amplification that is below the level of detection of the real time instrument. Threshold – where the threshold and the amplification plot intersect defines CT. Can be set manually/automatically CT – (cycle threshold) the cycle number where the fluorescence passes the threshold Rn – (Rn-baseline) NTC – no template control Rn is plotted against cycle numbers to produce the amplification curves and gives the CT value. ONE-STEP OR TWO-STEP REACTION qRT-PCR can be one step or two step. 1. Two-step qRT-PCR Two-step qRT-PCR starts with the reverse transcription of either total RNA or poly(A)+ RNA into cDNA using a reverse transcriptase (RT). This first-strand cDNA synthesis reaction can be primed using random hexamers, oligo(dT), or gene-specific primers (GSPs). To give an equal representation of all targets in real-time PCR applications and to avoid the 3? bias of oligo(dT), it is usually recommended that random hexamers or a mixture of oligo(dT) and random hexamers are used. The temperature used for cDNA synthesis depends on the RT enzyme chosen. Following the first-strand synthesis reaction, the cDNA is transferred to a separate tube for the qPCR reaction. In general, only 10% of the first strand reaction is used for each qPCR. . One-step qRT-PCR One-step qRT-PCR combines the first-strand cDNA synthesis reaction and qPCR reaction in the same tube, simplifying reaction setup and reducing the possibility of contamination. Gene-specifi c primers (GSP) are required. This is because using oligo(dT) or random primers will generate nonspecific products in the one-step procedure and reduce the amount of product of interest. O verview of qPCR and qRT-PCR components This section provides an overview of the major reaction components and parameters involved in real-time PCR experiments. * DNA polymerase One of the main factors affecting PCR specificity is the fact that Taq DNA polymerase has residual activity at low temperatures. Primers can anneal nonspecifically to DNA, allowing the polymerase to synthesize nonspecific product. The problem of nonspecific products resulting from mispriming can be minimized by using a â€Å"hot-start† enzyme. Using a hot-start enzyme ensures that no active Taq is present during reaction setup and the initial DNA denaturation step. * Template Anywhere from 10 to 1,000 copies of template nucleic acid should be used for each real-time PCR reaction. This is equivalent to approximately 100 pg to 1 ? of genomic DNA, or cDNA, generated from 1 pg to 100 ng of total RNA. Excess template may increase the amount of contaminants and reduce efficiency. If the template is RNA, care should be taken to reduce the chance of genomic DNA contamination. One option is to treat the template with DNaseI. Ultrapure, intact RNA is essential for full-length, high-quality cDNA synthesis and accurate mRNA quantification. RNA should be devoid of any RNase contamination, and aseptic conditions should be maintained. * Reverse transcriptase The reverse transcriptase (RT) is as critical to the success of qRT-PCR as the DNA polymerase. It is important to choose an RT that not only provides high yields of full-length cDNA but also has good activity at high temperatures. High-temperature performance is also very important for tackling RNA with secondary structure or when working with gene-specific primers (GSPs). * dNTPs It is recommended that both the dNTPs and the Taq DNA polymerase be purchased from the same vendor, as it is not uncommon to see shifts of one full threshold cycle (Ct) in experiments that employ these items from separate vendors. * Magnesium concentration In qPCR, magnesium chloride or magnesium sulfate is typically used at a fi nal concentration of 3 mM. This concentration works well for most targets; however, the optimal magnesium concentration may vary between 3 and 6 mM. * UNG The Uracil-N-Glycosylase is an enzyme that hydrolyses all single-stranded and double-stranded DNA containing dUTPs. Consequently, if all PCR amplifications are performed in the presence of a dNTPs/dUTPs blend, by carrying a UNG step before every run it is possible to get rid of any previous PCR product. * ROX Some thermocyclers require MasterMix containing ROX dye for normalization. This is the case for the ABI and Eppendorf machines, and optional on the Stratagene machines. If you work with such machines, it is easier to work with the ROX dye already incorporated in the MasterMix rather than adding it manually. It guarantees a higher level of reproducibility and homogeneity of your assays. * Fluorescein For iCycler iQ, My iQ and iQ5 machines (BioRad thermocyclers), the normalization method for SYBR Green assay uses Fluorescein to create a â€Å"virtual background†. As in the case for the ROX, it is better and easier to use a MasterMix that contains pre-diluted Fluorescein, guaranteeing higher reproducibility and homogeneity of your assays. REAL TIME PCR SYSTEM: System Features: †¢ Four interchangeable block formats †¢ Optional Automation Accessory amp; Barcode Scanner †¢ Argon ion laser/CCD camera †¢ Easy to Use Software, Multiple Applications †¢ Set up Wizards †¢ QC Filtering/Flag System †¢ Flexible data reports amp; exporting SOFTWARES FOR DATA ANALYSIS AND PRIMER DESIGNING 1 ) Light Cycler ® Relative Quantification Software The first commercially available software was the Light Cycler ® Relative Quantification Software (2001). 2 ) REST In 2002, the relative expression software tool (REST ) was established as a new tool. 3 ) Q-Gene Recently a second software tool, Q-Gene, was developed, which is able to perform a statistical test of the real-time data. Q-Gene manages and expedites the planning, performance and evaluation of quantitative real-time PCR experiments. 4) OligoPerfect A primer design software program such as OligoPerfectâ„ ¢, available on the Web at www. invitrogen. com/oligoperfect, can automatically evaluate a target sequence and design primers for it based on the criteria STEPS OF REAL TIME PCR Real-time reaction mix (final concentrations): 1x 2 x AmpliTaq Gold 0. 5 ? M 5’ primer 0. 5 ? M 3’ primer 0. 2 ? M probe 0. 4 ? Rox reference dye 20 ? l Final Volume (including sample and dH20) STANDARD REAL-TIME PCR PROTOCOL ASSAY DESIGN: This section describes the stages of real-time PCR assay design and implementation. We will identify sources of variability, the role they play in data accuracy, and guidelines for optimization in the following areas: 1Target amplicon and primer design 2. Nucleic acid purification 3. Reverse transcription 4. Controls and normalization 5. Standard curve evaluation of efficiency, sensitivity, and reproducibility Good primer (pair) properties One way to minimize efficiency bias is to amplify relatively short targets. Amplifying a 100 bp region is more likely to result in complete synthesis in a given cycle than, say, amplifying a 1,200 bp target. For this reason, real-time PCR target lengths are generally in the range of 60 bp to 200 bp. In addition, shorter amplicons act as a buff er against variations in template integrity. Primers designed to amplify larger regions are less likely to anneal with the same fragment in a slightly degraded nucleic acid sample. PURIFICATION Phenol-based organic extraction is a very effective method for purifying RNA from a wide variety of cell and tissue types. During sample lysis, phenol and guanidine isothiocyanate disrupt cells and dissolve cell components. while maintaining the integrity of the nucleic acids by protecting them from RNases. Chloroform is added and the mixture is separated by centrifugation, which separates the solution into an aqueous phase and an organic phase. RNA remains exclusively in the aqueous phase in the presence of guanidine isothiocyanate, while DNA and protein are driven into the organic phase and interphase. The RNA is then recovered from the aqueous phase by precipitation with isopropyl alcohol. REVERSE TRANSCRIPTION CONSIDERATIONS Most reverse transcriptases employed in qRT-PCR are derived from avian myeloblastosis virus (AMV) or Moloney murine leukemia virus (M-MLV). An ideal reverse transcriptase will exhibit the following attributes: * Thermostability— thermostable RTs function at the higher end of (or above) this range and allow for successful reverse transcription of GC-rich regions. * RNase H activity— RNase H activity can drastically reduce the yield and ratio of full-length cDNA, which translates to poor sensitivity. Several RTs, most notably SuperScript II and III, have been engineered for reduced RNase H activity. NORMALIZATION AND QUANTIFICATION: When analyzing and comparing results of Real-Time qPCR assays many researchers are confronted with several uncontrolled variables, which can lead to misinterpretation of the results. Those uncontrolled variables can be the amount of starting material, enzymatic efficiencies, and differences between tissues, individuals or experimental conditions. In order to make a good comparison, normalization can be used as a correction method, for these variables. The most commonly known and used ways of normalization are : * normalization to the original number of cells, * normalization to the total RNA mass, normalization to one or more housekeeping genes, * normalization to an internal or external calibrator. Normalization to number of cells can actually only be done for cell culture and blood samples. The two majors methods of normalization are the absolute quantification and the relative quantification . Absolute quantification Absolute quantification requires a standard curve of known copy numbers. The amplicon being studied can be cloned, or a synthetic oligonucleotide (RNA or DNA) can be used. The standard must be amplified using the same primers as the gene of interest and must amplify with the same efficiency. The standards must also be quantified accurately. This can be carried out by reading the absorbance at A260, although this does not distinguish between DNA and RNA, or by using a fluorescent ribonucleic acid stain such as RiboGreen. Relative quantification Relative quantification is the most widely used technique. Gene expression levels are calculated by the ratio between the amount of target gene and an endogenous reference gene, which is present in all samples. The reference gene has to be chosen so that its expression does not change under the experimental conditions or between different tissue. There are simple and more complex methods for relative quantification, depending on the PCR efficiency, and the number of reference genes used. STANDARD CURVE TO ASSESS EFFICIENCY, SENSITIVITY, AND REPRODUCIBILITY The final stage before assay employment is validating that all the experimental design parameters result in a highly efficient, sensitive, and reproducible experiment. * Reaction efficiency One hundred percent efficiency corresponds to a perfect doubling of template at every cycle, but the acceptable range is 90–110% for assay validation. This efficiency range corresponds to standard curve slopes of –3. 6 to –3. 1. The graph in Figure shows the measurement bias resulting solely from differences in reaction efficiency.. A standard curve is generated by plotting a dilution series of template against the Ct for each dilution. To some, sensitivity is measured by how early a target Ct appears in the amplification plot. However, the true gauge of sensitivity of an assay is whether a given low amount of template fits to the standard curve while maintaining a desirable efficiency. The most dilute sample that fits determines reaction sensitivity. The standard curve also includes an R2 value, which is a measure of replicate reproducibility. Standard curves may be repeated over time to assess whether the consistency, and therefore the data accuracy for the samples. Real-Time PCR Fluorescence Detection Systems Several different fluorescence detection technologies can be used for realtime PCR, and each has specific assay design requirements. All are based on the generation of a fluorescent signal that is proportional to the amount of PCR product formed. The three main fluorescence detection systems are: * DNA-binding agents (e. g. SYBR Green and SYBR GreenER technologies * Fluorescent primers (e. g. , LUX Fluorogenic Primers and Amplifluor qPCR primers) * Fluorescent probes (e. g. , TaqMan probes, Scorpions, Molecular Beacons) The detection method plays a critical role in the success of real-time PCR. DNA-Binding Dyes The most common system for detection of amplified DNA is the use of intercalating dyes that fluoresce when bound to dsDNA. SYBR Green I and SYBR GreenER technologies use this type of detection method. The fluorescence of DNA-binding dyes significantly increases when bound to double-stranded DNA (dsDNA). The intensity of the fluorescent signal depends on the amount of dsDNA that is present. As dsDNA accumulates, the dye generates a signal that is proportional to the DNA concentration and can be detected using real-time PCR instruments. SYBR Green I advantages †¢ Low cost assay †¢ Easy design and set up SYBR Green I disadvantages †¢ Non specific system †¢ Not adapted to multiplex †¢ Non suitable for qualitative qPCR Primer-Based Detection Systems Primer-based fluorescence detection technologies can provide highly sensitive and specific detection of DNA and RNA. In these systems, the fluorophores is attached to a target-specific PCR primer that increases in fluorescence when incorporated into the PCR product during amplification. * Amplifluor Real-Time PCR Primers Amplifluor real-time PCR primers are designed with both a fluorophore and quencher on the same primer. The primer adopts a hairpin configuration that brings the fluorophore in close proximity to the quencher. The fluorescent signal increases when the primer is unfolded and the fluorophore and quencher are de-coupled during incorporation into an amplification product. Figure: Ampliflour primer PROBE-BASED DETECTION SYSTEMS Probe-based systems provide highly sensitive and specifi c detection of DNA and RNA. However, dual-labeling and complex design specifi cations make them expensive and more diffi cult to use than primer-based systems or DNAbinding dyes. TaqMan probes = Double-Dye probes TaqMan probes, also called Double-Dye Oligonucleotides, Double-Dye Probes, or Dual Labelled probes, are the most widely used type of probes. A fluorophore is attached to the 5’ end of the probe and a quencher to the 3’ end. The fluorophores is excited by the machine and passes its energy, via FRET (Fluorescence Resonance Energy Transfer) to the quencher. TaqMan probes can be used for both quantification and mutation detection, and most designs appear to work well. TaqMan ASSAY DENATURATION ANNEALING OF PRIMERS AND PROBE POLYMERIZATION AND PROBE CLEAVAGE Molecular Beacons In addition to two sequence-specific primers, molecular beacon assays employ a sequence-specific, fluorescently labeled oligonucleotide probe called a molecular beacon, which is a dye-labeled oligonucleotide (25–40 nt) that forms a hairpin structure with a stem and a loop . A fluorescent reporter is attached to the 5 end of the molecular beacon and a quencher is attached to the 3 end. The loop is designed to hybridize specifically to a 15–30 nucleotide section of the target sequence Figure: Moleculer Beacon They are highly specific, can be used for multiplexing, and if the target sequence does not match the beacon sequence exactly, hybridization and fluorescence will not occur a desirable quality for allelic discrimination experiments. Hybridization probes (also called FRET probes) Roche has developed hybridization probes for use with their LightCycler. Two probes are designed to bind adjacent to one another on the amplicon. One has a 3’ label of FAM, whilst the other has a 5’ LC dye, LC red 640 or 705. When the probes are not bound to the target sequence, the fluorescent signal from the reporter dye is not detected. However, when the probes hybridize to the target sequence during the PCR annealing step, the close proximity of the two fluorophores allows energy transfer from the donor to the acceptor dye, resulting in a fluorescent signal that is detected. FRET probe principle and light cycler MELTING CURVE ANALYSIS Melting curve analysis can only be performed with real-time PCR detection technologies in which the fluorophore remains associated with the amplicon. Amplifications that have used SYBR Green I or SYBR GreenER dye primers can be subjected to melting curve analysis. Dual-labeled probe detection systems such as TaqMan probes are not compatible because they produce an irreversible change in signal by cleaving and releasing the fluorophore into solution during the PCR; however, the increased specificity of this method makes this less of a concern. The level of fluorescence of both SYBR Green I and SYBR GreenER dyes significantly increases upon binding to dsDNA. By monitoring the dsDNA as it melts, a decrease in fluorescence will be seen as soon as the DNA becomes single-stranded and the dye dissociates from the DNA. Figure: Melting curve analysis can detect the presence of nonspecifc products, as shown by the additional peaks to the left of the peak for the amplified product in the melt curve. How to perform melting curve analysis To perform melting curve analysis, the real-time PCR instrument can be programmed to include a melting profile immediately following the thermocycling protocol. After amplification is complete, the instrument will reheat your amplified products to give complete melting curve data. Most real-time PCR instrument platforms now incorporate this feature into their analysis packages. In general, the program steps will be: 1. Rapid heating of the amplified sample to 94 °C to denature the DNA. 2. Cooling the sample to 60 °C. 3. Slowly heating (by increasing the temperature 0. 2 °C/second) the sample while plotting fluorescence signal vs. temperature. (As the temperature increases and the dsDNA strands melt, the fluorescence signal will decrease. ) Figure: Example of a melting curve thermal profile setup on an Applied Biosystems instrument (rapid heating to 94 °C to denature the DNA, followed by cooling to 60 °C. ) Multiplex real-time PCR In multiplex real-time PCR, more than one set of gene-specific primers is used to amplify separate genes from the template DNA or RNA in a single tube. Typically, multiplex reactions are used to amplify a gene of interest and a â€Å"housekeeping† gene (e. g. , #-actin or GAPDH), which is used as a normalize for the reaction. Because more than one PCR product will be quantified in the same tube, different fluorescent reporter dyes are used to label the separate primers or probes for each gene. More Samples Analyzed per Plate. Target and normalizer in same reaction and Less sample consumed. APPLICATIONS OF REAL TIME PCR GENE EXPRESSION ANALYSIS A sample gene expression analysis using a multiplex TaqMan assay is presented in the following sections. In this example, we’re interested in the relative expression of three genes in the polyamine biosynthesis pathway, ornithine decarboxylase (ODC), ODC antizyme (OAZ), and antizyme inhibitor (AZI), in two different samples, sample A and sample B. 1. RNA was isolated from sample A and sample B. 2. RNA was reverse transcribed into cDNA. 3. The amount of the target genes (ODC, OAZ, and AZI) and the reference gene (b-actin) was determined in each of the cDNA samples using a multiplex qPCR assay. 4. Data were analyzed and the relative expression of each of the target genes in the two samples was calculated. EXAMPLE BRCA1 is a gene involved in tumor suppression. BRCA1 controls the expression of other genes. In order to monitor level of expression of BRCA1, real-time PCR is used. SNP GENOTYPING In order to perform SNP genotyping, two specific probes labeled with different dyes are used, the first for the wild type allele and the second for the mutant allele. If the assay results in the generation of only the first fluorescent color, then the individual is homozygous wild type at that locus. If the assay results in the generation of only the second fluorescent color, then the individual is homozygous mutant. And finally, if both fluorescent colors are produced, then the individual is heterozygous. At the end of the reaction, hydrolysis probes are digested. The quality of a hydrolysis probe is given by the hybridization efficiency, the quenching of the intact probe and the cleavage activity of Taq polymerase. HIV DETECTION Nowadays HIV is strikingly spreading out whole the world. so in order to diminish its distribution , it is necessary to detect it as soon as possible amp; for this purpose, Real time PCR is recommended by scientist. In this method ,’ pol’’ gen of the virus, is amplified in thermocycler. 6 patient have been studied. infection in these patients was confirmed by ELISA amp; western blot. * Sampling amp; RNA extracting from patients. * Cloning of target segment by using Xba I amp; Hind III. And 180 bp primers. * Standard virus mRNA was extracted. * Quantitative analysis of HIV virus by SYBR-green Real Time RT-PCR. CYSTIC FIBROSIS (CF) DETECTION: Cystic f ibrosis (CF) is the most common inherited disease among Caucasian populations with an incidence of ~1 in 2500 births. A3 base pair (bp) deletion, designated DF508, accounts for nearly 70% of CF cases and causes severe manifestations of the disease. It results in the absence of phenylalanine at position 508 of the cystic fibrosis transmembrane conductance regulator (CFTR) protein and this error prevents normal processing and translocation of the polypeptide chain to apical membranes of epithelial cells. This deletion can be detected by molecular beacons in real time PCR. Figure:Examples of specific molecular beacon fluorescence increase during real-time PCR in samples containing single lymphoblasts homozygous normal for CF (green), heterozygous DF508 (blue), or homozygous DF508 (red). A) Fluorescent signal from the molecular beacon detecting the normal allele. (B) Fluorescent signal from the molecular beacon detecting the DF508 allele. Dashed lines indicate the threshold of 200 units (~10 SD above baseline readings) used for determining CT values. THE ADVANTAGES OF REAL-TIME PCR * The ability to monitor the progress of the PCR reaction as it occurs in real time * The ability to precisely measure the amount of amplicon at each cy cle * An increased dynamic range of detection * The combination of amplification and detection in a single tube, which eliminates post-PCR manipulations. Rapid cycling times (1 hour) * High sample throughput (~200 samples/day) * Low contamination risk (sealed reactions) * Very sensitive (3pg or 1 genome eq of DNA) * Broad dynamic range (10 1010 copies) * Reproducible (CV lt; 2. 0 %) * Allows for quantitation of results * Software driven operation * No more expensive than â€Å"in house† PCR ($15/test) THE DISADVANTAGES * Current technology has limited capacity for multiplexing. Simultaneous detection of 2 targets is the limit. * Development of protocols needs high level of technical skill and/or support. Requires Ramp;D capacity and capital) * High capital equipment costs ($ 50,000 -160,000). REFRENCES * http://www. icmb. utexas. edu/core/DNA/qPCR/QiagenRT-PCR. pdf www. icmb. utexas. edu * http://books. google. com. pk/books? id=-v-U-mXWg-gCamp;printsec=frontcoveramp;dq=real+time+pcramp;hl=enamp;sa=Xamp;ei=Bph1UezKIceDhQeUh4CwCAamp;ved=0CDAQ6AEwAQ#v=onepageamp;q=real%20time%20pcramp;f=false books. google. com. pk * PCR/Real-Ti me PCR Protocols www. protocol-online. org Real-Time Pcr: An Essential Guide Google Books books. google. com * * http://www. gene-quantification. e/bio-rad-CFX96-bulletin-5589. pdf www. gene-quantification. de * https://www. google. com. pk/#output=searchamp;sclient=psy-abamp;q=fret+rt-qpcramp;oq=fret+in+rtamp;gs_l=serp. 1. 1. 0i22i30l2. 1583. 4622. 1. 10196. 6. 6. 0. 0. 0. 0. 551. 2584. 3-3j1j2. 6. 0 0. 0 1c. 1. 9. serp. 97Wjtm9UCU4amp;psj=1amp;bav=on. 2,or. r_cp. r_qf. amp;fp=f6d28cf5fd703914amp;biw=1366amp;bih=600 www. google. com. pk * BioTechniques Real-time PCR for mRNA quantitation www. biotechniques. com * http://env1. gist. ac. kr/joint_unugist/file/g_class11_real_time_pcr_vt. pdf env1. gist. ac. kr

Letter to Newlyweds free essay sample

Letter to Newlyweds Rosemarie Velez Com200: Interpersonal Communication Prof. Megan Pope March 25, 2013 Dear Newlyweds, It has come to my attention that you have been seeking advice to keep the love alive throughout your marriage. I may say I am no expert but my research has given me the advantage to help you out. As a couple you should practice ways to keep your marriage alive; managing interpersonal conflicts, learning the importance of listening, and learning how to intact your emotions and speak to one another is what will help you understand one another. I will section these main points for you so that you will understand more. Managing interpersonal conflicts The reason being that it is important to manage interpersonal conflicts is because you want to make sure that issues are being resolved, you never want to have a disagreement about something and never come to a common ground in the situation it will only cause tension and make things worse. We will write a custom essay sample on Letter to Newlyweds or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Based on a website I have found that gives relationship advice, it is said, â€Å"People have varying degrees of comfort with conflict. Some prefer avoiding it at all costs. Unfortunately, those costs tend to increase the longer issues are left unaddressed. Therefore, learning how to manage and resolve conflict is to your benefit†. (Life Tips, 2013). From this statement it is clear that there are couples who put very serious issues in the back burner and when that occurs the issue becomes long term. That is why you must learn how to manage your conflicts and instead or yelling at each other it is best that you speak calmly. Something else that involves managing your interpersonal conflicts includes understanding the other person. â€Å"Attempt to understand the other persons point of view. Dismissing the others views, assigning blame, and exclusive focus on your own perspective are all counterproductive. † (Life Tips, 2013) When communicating with your partner about something it is clear that your feelings are not the only thing that matters. You must be able to put your views aside and listen and try to understand why your partner feels the way they feel. Being understanding is a step to making things better because once you both understand the others point of view than you can come together as one and have a common ground on the issue. Develop strategies for active, critical, and empathic listening When communicating with your partner is listening, not hearing, listening. Some partners say that they are listening when they really aren’t. When talking things out paying attention to what your partner is saying or how they are feeling is important. The reason why it is so important because when you are not listening to your partner it becomes an issue. â€Å"One of the most neglected interpersonal communication skills, and a core competence we must master to be an effective communicator, is listening. You cannot understand others, respond appropriately to what they say, and provide helpful feedback if you have not listened†. (Sole, 2011) This is a statement that completely backs up to what I just said. Listening is a big factor in communication in a marriage or in any relationship in that matter. You have to give one hundred percent to keep that relationship healthy. Listening is the act of hearing and interpreting the message being conveyed from another person. Many couples get together and stop listening to one another. All they do is hear. Hearing is not enough if a couple wants to remain together. (Diggs, Angela, 2009) In this statement that is found in an article that was posted on Helium. com Diggs stated four reasons why listening is important and they were; keeps relationships together, helps develop intimacy in connection, shows respect and healing to relationships. I hope that these four reasons why listening is important can help you two to always listen to one another. Emotional intelligence and its role in effective interpersonal relationships Being able to control your emotions in an effective interpersonal relationship means controlling that moment where information is given that must be clear with emotions not getting out of control. An example I have found on cbtrecovery. com states, â€Å" · In relationships: Don’t let hurts and problems build up. Examples? How can you prevent problems from getting worse? Resolve conflicts before they get overwhelming. End hopeless relationships. † Being that you two are newlyweds your marriage is fresh everything is perfect. From what I have heard from people that are married things change over time; kids are made, a house is bought, things become more stressful and both of you will have new responsibility that you both have to take care of together as a team. I know that as women we can get very emotional about everything and as a man emotions will not be shown because it is ignored. So as a husband and wife you should learn how to intact your emotions so that it is shown but not shown too much. As a married couple you have to be able to communicate all the time even if it’s just talking about at favorite movie or the activity you did or even how work went. Doing that can help enhance your communication and if or when there is something that is bothering the other person it will be talked about. An example I have that may help you is my mom’s marriage. Her communication with her husband was very poor so when issues aroused it became very emotional for her because she took her time speaking about it. When that happens it can cause confusion it can even cause misunderstanding in the relationship which is not very good. My mom end up learning that and now she is separated. I always say communication is key to a healthy relationship and from the information I have given you so far it seems true; especially for marriage. This is a statement from a doctor and it explains the different emotional types of people. â€Å"In healthy, constructive relationships, it’s no secret that solid communication skills and the ability to process your emotions always take a front-row seat. But did you know that in the world of relationships, there are two different types of people? You’re either an emotionally responsible person or an emotionally dependent one. Emotional responsibility is very important in a healthy, satisfying relationship. It facilitates personal growth, as well as growth between you and your partner†. (Dr. Laura Berman) Recognize how words have the power to create and affect attitudes, behavior, and perception The way you speak to your partner and the way you act is a really big thing that causes issues in your relationship. If you and your partner are in a disagreement and one is yelling at the other it tends to cause an emotional imbalance within the person. Meaning that the emotions are going crazy to the point the person doesn’t understand why they are being yelled at. In my own experience I have had moments where as my partner would get upset about something and speak to me in a disrespectful manner and when I would try to speak to him to see what was wrong his body language and facial expression would show his aggravation. When that would happen my emotions would erupt and I would not understand what was going on that caused the situation to get worse because either we would ignore everything and it would build up or we would just argue and never fix things. It’s important that when you are talking with your partner that you are being respectful you are listening you are showing that you are listening instead of having wandering eyes. This can help your communication get better because you will know how to control the words you choose and learn how to have the write body language and facial expression. The words you choose can make your relationship a lot better, or at times, it can push your partner away from you forever. You may communicate well, share your dreams, respect your lover and even be great in bed, but the way you use the power of words could still affect your relationship on a subconscious level. † (Avdeeva, Natalia, 2012) This statement supports what I have stated. The proce ss by which self-concept is developed and maintained In your relationship there are things that you both have to do to be at peace with yourself. In marriage things can be perfect as a team but sometimes things aren’t always perfect within you. So that’s where self-concept comes in and I will give you examples of how it can be maintained. Self-concept is the way you look at yourself it’s a judgment that you have of yourself. If you look at yourself in a negative way it can cause your partner to look at you the same way. You want to always see and think of yourself in a positive way regardless of what. If you feel ugly and your partner finds you’re the most beautiful thing on earth than you believe it. The way you look, your self-esteem and self-worth all should be positive. I know for women our self-image can mean so much that will cause our self-esteem low but when being in a relationship you have to be positive about yourself that shows that you are confident in yourself in your marriage and a person that won’t let things ruin anything. â€Å"Developing a healthy self-concept takes deliberate planning and concentrated effort. It takes acknowledging your intrinsic value as a human being, and then working to acquire the skills needed to confront the many challenges and adversities we encounter in life. When you possess a healthy self-concept, nothing can rattle you, or take you off your stride. You are confident, poised, and assured because you know you are equipped to handle whatever comes your way. † (Essential Life Skills) A healthy self-concept includes knowing yourself, assessing your strengths and weaknesses as well as knowing your potential to do things. You should always love and accept who you are, be true to who you are and take responsibilities for the decisions you make. To maintain these healthy ways to self-concept is to be positive as I said before. It also helps to have positive people around you as well. That should be able to help you with who you are and how you view each other as one. Now don’t ever get discouraged if you have disagreements about something just remember the importance of interpersonal communication as a married couple. As newlyweds this is the time to get to know each other more, on a deeper level. The advice I have given you is to get you started and to continue to keep your marriage healthy and happy. Controlling your interpersonal conflicts, listening and understand each other, and speaking to each other the right way where emotions and being thought of is what is going to help you, not only as a married couple but as friends and as people. References http://relationship. lifetips. com/cat/64813/managing-conflict-in-relationships/index. html Sole, K. (2011). Making connections: Understanding interpersonal communication. San Diego, CA: Bridgepoint Education, Inc. Schoenberg, N. (2011, January 17). Can we talk? Researcher talks about the role of communication in happy marriages. McClatchy-Tribune News Service. Retrieved from ProQuest http://www. drlauraberman. com/relationships/committed-love/emotional-responsibility

Observing Stars Essays - Electromagnetic Radiation, Radiation

Observing Stars Essays - Electromagnetic Radiation, Radiation Observing Stars Observing Stars Our view of the sky at night is possible because of the emission and reflection of light. 'Light' is the better-known term for the electromagnetic spectrum, which includes waves in the visible, ultra-violet, infra-red, microwave, radio, X-ray and gamma-ray regions. The scale of the spectrum is so large that no region is distinct, several overlap each other. Each of these regions in the electromagnetic spectrum represent transverse waves, travelling as electrical and magnetic fields which interact perpendicularly to each other, with different ranges of wavelength. The magnetic field oscillates vertically and the electric field horizontally, and each field induces the other. By the end of the nineteenth century, Maxwell gave a realistic value for c, the speed of light: c = __1__ = 3 x 108 ms-1 (mo eo) The relationship between the speed of all electromagnetic radiation, wavelength (l) and frequency (f) is shown to be c = l f. Because the Universe is so vast, interstellar distances are so great that light emitted can take upwards of millions of years to reach us. Such large distances are often measured in light-years; one light-year (ly) is the distance travelled by a wave of light in a year. Because of the massive speed of light and distances, the light arriving at us would have left the object many years ago, so that looking at a far away star is much like looking back in time. Scientific observation of the stars is difficult because of the distorting effect of the Earth's atmosphere. One problem is atmospheric refraction-where light is bent. Turbulent air currents cause varying refractive indices, as there is no uniform air density. This causes an effect called scintillation, where stars appear to twinkle. The effect on regions of the electromagnetic spectrum other than the visible part, such as the absorption of certain frequencies by atmospheric chemicals, and the reflection of waves by charged molecules in the ionosphere, means that some spectral data is simply invisible to us on Earth. The Earth receives electromagnetic radiation of all wavelengths from all directions in space, but most of the electromagnetic spectrum is blocked out by the atmosphere well above the Earth's surface, where our eyes and instruments are mostly based. However, wavelengths from only two regions of the electromagnetic spectrum are able to penetrate the atmosphere. These two spectral windows in the atmosphere through which we can observe the Universe are called the optical window-which allows the visible wavelength region through; and the radio window-which includes the wavelength region from about 1 mm to 30 m. The telescopes used by astronomers on the ground are therefore classed as optical and radio telescopes. Optical telescopes work by either reflecting or refracting light, using lenses or curved mirrors to focus the light from a subject to form an image. Radio telescopes consist of a parabolic reflector and receiver on which the waves are focused. The gathering and resolving power de pend on the diameter of the antenna. Radio observations are unaffected by the weather or time of day, and because of the larger wavelength of radio waves, dust in space and atmospheric convection currents are not a problem. Radio astronomy is used in the chemical analysis of elements (by emission and absorption spectra); to detect the motion of bodies due to the Doppler effect; and in investigation into the early Universe and the Big Bang. We can analyse radio waves from the centres of galaxies, including our own. Despite the radio window, there are still wavelengths that do not penetrate the atmosphere. Some radio waves are reflected from the ionosphere, part of the thermosphere, where streams of charged particles from the sun ionise gas molecules: this is photo-ionisation. Ultra-violet radiation, X-rays and gamma-rays are also absorbed at this layer. Absorption of the electromagnetic spectrum at various altitudes above Earth occurs to varying degrees. Much infra-red radiation does not reach ground level because of absorption in the upper atmosphere by water, and some carbon dioxide and oxygen molecules that lie between the ground and about 15 km of altitude (the troposphere). Ozone (tri-oxygen) and di-oxygen in the stratosphere absorbs much of the ultra-violet radiation (hence the ozone layer at about 30km). A side effect of the ozone layer is that molecules re-radiate

War on Ignorance Essay Example | Topics and Well Written Essays - 750 words

War on Ignorance - Essay Example He punctuates his lamentations with a litany of what illiterates cannot do: "Illiterates cannot read the menu at a restaurantcannot look up numbers in a telephone directorycannot read the notices they receive from welfare offices" (284). Kozol puts some of the blame on the parents of the past who neglected their duty to educate themselves, passing off to their illiteracy as an heirloom to their offspring, misery begetting more misery when human survival in our society is almost impossible without the mental ability needed to do so. He, however, puts most of the blame on the government and the education officials whose job it is to ensure that as many as possible should benefit in our democratic and wealthy society, crying that "so long as 60 million Americans are denied significant participation, the government is neither of, nor for, nor by the people" (286). After laying out the costs paid both by those who suf... Barry's short essay, while contrasting with Kozol's polemics, even more dramatically points out a deeper problem in society that is related to our thesis that America is losing its mental edge: parents who don't care about their family because they are too busy taking care of a thousand and one details about their family (308). Yes, this seems like a chicken-and-egg argument, but Barry brings home the point that in an America where working hard and smart is the norm, children are not getting the encouragement to prepare themselves for the future. We can only lose if we continue neglecting our children. This is not to say that Barry was not motivated to go to school nor, like Stephen in Kozol's essay, she was illiterate. She was intelligent and smart, and her opening statements revealed it: School "was a world I absolutely relied on. Without it, I don't know where I would have gone that morning." (307). School was her sanctuary, and it was probably what gave her and her brother the sense to do better than their parents and get out of trouble. In a way, Barry is more optimistic because school for her became the home she yearned for, a safe and secure haven where people cared and where her teacher, Mrs. Lesane, gave love to those who needed it (308). She showed that if America's teachers are dedicated as her teacher, many children who would otherwise be lost to ignorance can be won over and recovered and can grow up to be happy, creative, and useful citizens who can contribute to making this country great. Like Kozol, Barry directs her appeal to parents not to neglect their children. Lynda's parents were lucky, because she and her brother were smart, so they turned out alright. Her essay, however, leaves us wondering what

Emplyment Relation Essay Example | Topics and Well Written Essays - 1000 words

Emplyment Relation - Essay Example Another problem which is aggravating the situation is the arrangement of finances for the back-pays and the future wage increases by the councils. The dispute resolution process can be divided into three phases based on the types of grievances. All the three phases will be at work simultaneously with three separate HR teams working on the assignment. The first phase will address the issues of the employees who have been subjected to salary cuts. It is apparent that no employee will accede to their salary cut on the pretext of social justice. However, the financial prudence of implementing the equal pay policy requires that some financing of hike in wages be done through some salary cuts. So, the first thing that can be done is to re-evaluate the grades of those employees who have been subjected to salary cuts. This will be a time consuming exercise, however it will help in avoiding any further litigation. Some time can be saved by looking at data from previous litigations by such employees. It has been observed that some particular jobs had been upgraded after going into litigation. This data can be helpful in re-grading. A ta rget time can be set for this phase and during this time the salaries and other employment terms remain unchanged. For those whose grades are upgraded after the re-evaluation exercise, arrears can be paid at a later agreed-upon time. With this step the problems of the most aggrieved section of employees can be addressed. The time period for this re-evaluation of grades should not be more than 9 months. For those whose salaries will be upgraded after the re-grading exercise, there will be appropriate salary raise. For others whose grades still need to be revised towards the lower side, there will be no reduction in salary but only grade change. There other benefits will be downgraded but not the salary. They will be given a signing bonus in the form of 2.5% of their salary increment. These

Wal-Mart Business aroung the World Essay Example | Topics and Well Written Essays - 750 words

Wal-Mart Business aroung the World - Essay Example As the paper highlights, Mexico was not a driving region so they have to be within their consumers’ neighborhood. Aside from that, Mexicans preferred fresh products so they had to offer their goods fresh. With their thirst to succeed in their endeavor and the help of Grupo Cifra, Mexico became their first international expansion in 1991.   Aside from big established retailers like Metro AC, Aldi and Kaufland, Wal-Mart’s big mistake in their venture in Germany was not having a cultural context on their strategy. Same thing with their business in South Korea, Wal-Mart seemed to cater to Americans alone and did not mind the region’s culture. Unlike with their business in Mexico, they didn’t try to adapt to the needs and wants of their consumers. They eventually had to pull out their business in these countries. Offering freshly harvested products is a must for Chinese. Given that China shares 1/3s of Asia’s population, Wal-Mart considers China as the center of their growth strategy.   International business venture is never easy. There are so many factors to consider. Culture is on top of the listr. As a retailer, you should know the goods a region will buy to stay in business. A rigorous supply and demand and shopping habits study will be essential in order to succeed in your venture. Remem ber that to stay in business, you should get a good percentage of the market. Wal-Mart posted their third quarter profits reaching $3.23 billion, which is 3.2% from a year ago. Their net sales for the third quarter were $98.667, which increased 1.1% from the same period last year. Wal-Mart has more than 7,970 stores including Supercenters and discount stores.  

An analysis of eBay

An analysis of eBay INTRODUCTION 1.eBay is known all over the world as the leading online Marketplace (TM) which came in to life in 1995,eBay Inc, was founded by Pierre M. Omidyarand its headquartered in San Jose, California, TM (2002). eBay created a special platform for the sales of goods and services by a passionate village of individuals and business. On days there are millions of items across thousand of categories for sale on eBay just like Half.com, eBays site dedicated to fixed price trading. eBay allows trade on a Local, National and International basis, with customize site in marketing all over the world. eBay, together with its sub-visions, bring online marketplaces for the sale of goods and services, online payment services, and online communication offerings to a diverse community of individual and business in the USA and Internationally. The company operates in three Intentions: eBay Marketplace, Payments, and Communications. The eBay Marketplaces: This platform provides infrastructure to enable online commerce in a variety of formats, including the traditional auction platform; and its other online platforms, such as Rent.com, Shopping.com, Kijiji, mobile.de, and Marktplaats.nl. Its services include trust and safety programs, reply forum, safe chanel program, eBay standard purchase protection program, customer support, tools and services. The Payments segment: Feeds a product for small businesses, online sellers, and individuals that enables them to send and receive payments online. Its services include joining the network, crosscheck of its PayPals account holders, withdrawing funds. The Communications segment: provide voice over Internet protocol calls between Skype subscribers, also provides connectivity to traditional fixed-line and handheld telephones. The company also provide online apartment rental services and comparison shopping resource service, as well as provides an Internet payment platform that allows merchants to process, and manage online payments. 2. MISSION STATEMENT OBJECTIVE eBay pioneers communities built on commerce, sustained by trust, and inspired by opportunity. eBay brings together millions of people every day on a local, national and international basis through variety of websites that focus on commerce, payments and communication. This has also help the company in various ways between their numerous customers worldwide. They have engaged in the method stated below. A Place to Buy. You can get almost anything item you need or want at very low deal better than you can find at any traditional brick and mortar, even online stores. But caution should be taken because of rotten or fake items deals on eBay. A Place to Sell. No matter the size of our product or services, eBay has various categories channels to sell items. eBays global reach can even move unusual items that are not in demand in our own neighborhood in to cash. A Place to Shop. Large variety of items can be found for sale on eBay, many members have discovered that eBay is one of the best place in the world to window or comparison shop. These items include photos, detailed descriptions, and owner experiences. Because you see lots of the same items side by side in various conditions and know what each one sold or selling for, eBay gives you insight into the real market value of most types of goods around the globe. A Website:With no physical building eBay store, Founded in San Jose but now carry out daily activities from various cities, eBays service exist basically online apart from the delivery of bought and sold items every other network of the business are handled through eBay website. Socially Responsible: eBay with its B2C way of handling business has lower the barriers to buying and selling, these bring a massive global awareness to it business. It also fosters new economic opportunities to developing areas and brings cultural understanding between different populations. This has made eBay one of the worlds most interesting and exciting trans-national ambassadors. 3.THE IMPACT OF DIGITAL MEDIA ON EBAY eBay is a cutting edge in commence by selling almost everything be it of what nature with its presence in every corners of the globe with no physical store, with limited staffs but satisfy it numerous customer worldwide. But how is this possible? Social Media Marketing: Recent addition to organizations plans. Integrated marketing communications is a practice organization follow to connect with their target markets. Integrated marketing communications coordinates promotional elements, advertising, personnel selling, public relations, publicity, direct marketing and sale promotion. Increasingly viral marketing campaigns are also grouped into integrated marketing communications for eBay. The growth of social media has impacted the way organizations communicate, the internet provide set of idea which enable persons to develop social and business platform, share knowhow and collaborate on project online. Cell Phones: Usage has also become a benefit for eBay media marketing. Today many cell phones have social networking capabilities, individuals are notified of any happenings on social networkingsites through their cell phones in real-time. This constant connection to social networking sites means products and companies can constantly remind and update followers about their capabilities, uses, importance, etc. Since cell phones are connected to social networking sites, advertisements are always in sight. Computers: The use of computer has come to stay in our daily life be it at home, offices, school, this is one of the basic tools used by eMarketers worldwide, eBay uses the help of computer since they dont physical store to carry out their daily business and this is done by logging into their website by typing in the URL address with an access from ISP provider. Once logged in, you see various categorizes of product, prices, description of items and options of payment of items bought. Buying of items online and shipping to every part of the globe is an everyday activity at just by the use of a computer, Newsweek (2004) Internet Marketing: eBay use the internet known as digital marketing, web marketing, and online marketing is the marketing product and services over the internet. Internet marketing is considered to be broad in scope because it not only refers to marketing on the Internet, but also includes marketing done via e-mail and wirelessmedia. Digital customer data and electronic customer relationship management systems are also often grouped together under internet marketing. This market joins the vast idea and technical background of the Internet, including design, development, advertising, and sales.Internet marketing also refers to the placement of media along many different stages of the customer engagement cycle through search engine marketing on specific websites. 3.1 THE USE OF THE INTERNET BY eBay The internet as change the landscape of the world in every way things are done in every way things are done globally. Never before have people been able to interact in such a cost effective and comfortable way. The business world as seen a better new of doing its daily activity, business now are done in very short span of time even without the presence of both party at the same place or time. Now it is possible for you to establish your own company with a very small financial investment or in some cases even without any start-up capital. eBay use the internet in carrying of its daily business to meet target market and satisfy its customer at a very low cost and maximizing profit. Information: People worldwide are using the internet on a regular basis, with email and search engines as the most popular services. Information is power. People are able to influence, direct, convince, educate and manipulate others through one single tool: The distribution of information. Email and discussion forums allow people to share their thoughts, ideas and experiences with other people from all corners of the world. Within a relatively short period of time the Internet and its communication channels will be part of our daily lives as are running water and electricity. Cost Effectiveness: The internet is by far the most effective communication tool. If you want to send a letter via conventional or so-called slow mail, it will cost at least around $1 assuming you restrict yourself to two single sheets of paper. Sending the same amount of information via email will be up to 100 times cheaper with immediate delivery. In addition to this, email with its numbers of advantages over the slow mailing system of delivery of messages, sending and get a reply on the spot. Online Auction: In this massive marketplace, eBay rules the auction sites. According to the Nelison Rating, eBay is among the top ten most trafficked sites on the internet. eBay leads the online auction industry with a more than 60 percent share of the market, its closest competitors, Yahoo! Auctions, is only half its size followed by Amazon.com, all these are done with the help of the internet and makes eBay smile to the bank on daily basis. Low Cost Operation: Running business requires tons of capital no matter the size of the outfit, the business has to pay for rental shops, offices, motor vehicle, staffs, before it can think of making its profit which is the primary idea of the business, but doing online business you cut down your expenses to a very little amount of capital and that is the edge eBay is use to run its operations and has really help the business since it does not have physical presence anywhere, it does its business around the globe and sell lots of items to its customers worldwide. The internet helps eBay in an immeasurable way of cutting down cuts of operations and get to all part of the world without even being there in presence. 4. INTERNET AS A MARKETING PUBLIC RELATIONS TOOLS Public relations is defined as a management function which identifies, establishes and maintains mutually beneficialrelationships between an organization and the public upon which its success or failure depends. Whereas advertising is aone-way communication from sender to the receiver, public relations considers multiple audiences and uses two way communication to monitor feedback and adjust both its messages and the organizations actions maximum benefit. The internet is used as a tool in Public Relations and has replaced the use of paper form of notifications and is carried out in method stated below. Tweeter: Social media is a great way to have a conversation with your market and manage connections with prospects, customer, bloggers. Even if you dont join Twitter you can monitor what people are saying company, trends, friends and products. This is quite useful from a marketing and PR standpoint. Tweeter has a search engine that let you post any information and can be linked to eBays platform. Facebook: Business firms states their URLs on cable Tv   commercials, were friends and followers can join them on Facebook. This is sharp method way of internet PR, youre a startup entrepreneur, you could start a group to help entrepreneurs connect and exchange ideas. Search to make sure one doesnt exist already in your area. Your friends on your facebook will have first hand information of what is happening around you and your business and will also pass to from friends to friends if even they are not yours. eBay is linked to facebook which acts as an indirect form of PR, Instant message could also take form by chatting on both platform. Email: This is the fastest, stress less and cheapest form of PR and e-marketing with the use of the internet, when searching for information or to joint an organization like eBay, Amazon you go through a process of filling and providing certain information about yourself and your preference requirement, all these information is kept in their database and at the end you provide your e-mail. The organizations use their database information to communicate with you, through this medium you are updated with the latest information relating to previous request. As you change your preference change it is also updated in their database. eBay use this form of PR to promote their product to you and information is sent to you in respect of delivery of the item you paid for. Website Feedback: Various website use a page known as website feedback as a form PR tool to give a comments, complain or report on your experience you may have after buying a product or trial of certain items in other for them to know how their customer feel about product bought from them. 5. EBAY SWOT ANALYSIS STRENGHT:The Company use the advantage of Customer Relationship Management (CRM). Both parties (Buyers and Sellers) subscribe with the company and information is collected by eBay on individuals. This is known as Business2Consumer (B2C) of their business. However the strong customer relationships are founded on a Consumer2Consumer (C2C) business platform, where strong business relationships occurs, for example where buyers and sellers leave feedback for each other for the next transaction to take place. WEAKNESSES: Organizations works painfully to overcome fraud. However the eBay model does leave itself open to a number of fraudulent activities. But the company act with the issue very quickly. These includes inferior goods being marketed to unknown and suspecting eBayers. Other forms of theft could include the redistribution of stolen goods. Make it known that fraud and theft are problems with outsiders not eBay. The weakness is that mischievous individuals can exploit the C2C business platform. Many companies, systems breakdowns could disturb the trading transaction of eBay on daily bases. eBay and Paypal have suffered shutdowns and total breakdown. As technology improves for the better weakness will be less an issue. OPPORTUNITIES: Acquisitions bring new business strategy openings. eBay has into agreement to buy off online telephone company Skype Technologies with a deal reported of about worth $2.6 billion. PC users use Skype talk to each other for free and make cut-price calls to mobiles and landlines once you have subscribe to the services. This will bring in more customers to the business and also make profit to continue to grow. THRATS:The world Internet brands, success attract competition. Overseas competitors competing in their home markets have the technical experience that could give them a stronger advantage over eBay. It came to the notice of eBay that it has met with other USA home Internet companies when trading overseas. For example, Yahoo! Stand out in the Asian market. Doing the same market is a threat. As with weaknesses above, the name is attacked by mischievous individuals. When e-mail sent to unknown eBayers disguards to come from eBay. Logos, and design of the pages looks just the same. However they are designed so that you input private information that the thieves can use to take passwords and identifications. Not all costs can controlled by eBay, example shipping charges and credit card charges. With fuel prices to rise, the customer suffers the increment of delivery and postal charges. This affect the total cost of auctioned items very expensive. When provider of credit card merchants like Visa or Mastercard decide a charge for online business transaction the total cost of the same items would increase with a negative effects on the final customer of the product and will make eBay loose some of its future consumer and will affect the profit and share value of the company. 6. CONCLUSION EBay is well aware that in order for the company to maintain its competitive advantage and make it sustainable in the long run, the company must take advantage of the changes at the corporate, organizational and business levels. EBay embraces the five building blocks which are applied randomly throughout eBays history: improvisation, co-adaption, regeneration, experimentation and time pacing as evident by the strategies discussed. The company also embraces the 10 principles of competing on the edge about strategy, organization and leadership. With all hands on deck EBay can go further if the managing, marketing department embrace more with its IT department by merging with more social media platform not just their website because social website break more distance with communities of friends of my friends. Security department should work more bringing down the risk of internet fraud, this is one area of online business transaction that turn people away from buying online, because nobody wants to their money to end up on the place after paying for an item or product.

Katherine Mansfield’s Her First Ball Essay examples -- essays papers

Katherine Mansfield’s Her First Ball 1 In Katherine Mansfield’s Her First Ball, Leila, the main character is an eighteen-year-old girl from the rural country who has recently moved into the city with extended family members. To Leila, everything was â€Å"so new and exciting† (4th paragraph) and she immediately begins her path from innocence to experience. I can empathize with her, but I don’t feel sympathy for her. She didn’t have a tragic experience, she a learning experience. Leila’s metamorphosis during the course of the story proves that she is a heroine who overcame the conflict in her journey. 2 The structure of the plot begins with a description of the 6setting, which is interesting because the setting is not that significant to story because if it were in another time and place it would be just as effective.7 The way she describes the car was as though she was describing the road to enlightenment, and the image she gave of sitting in her own corner of the cab is significant foreshadowing: â€Å"she sat back in her own little corner of it, and the bolster on which her hand rested felt like the sleeve of an unknown young man’s dress suit; and away they bowled, past waltzing lamppost and houses and fences and trees.† (1st paragraph) Leila has so many expectations and anticipations that leads to the climax, and is filled with5 symbolism and color that describes a vivid vision of a need for freedom and experience: â€Å"the road was bright on either side with moving fan-...