“One can survive everything nowadays, except death, and live down anything, except a good reputation.” Oscar Wilde said, during the time of great Plague where many people are dying instantly and he intended that now a day’s people die but the people will never forget what it is to be living with the infection. In the play Hamlet by William Shakespeare, death of the characters is also similar to a plague that spreads all across and eventually they pass away. Many human beings lose their lives each day as a result of their own self-centered wrong doing; there are others whose deaths are an end result of manipulation from the royalty. The authentic tragedy of Hamlet is not that of Hamlet or his own family but of Polonius’ family deaths because they sacrificed their lives and were the innocent who were involved in order to make justice for king Hamlet. Sacrifice might end suffering but, causes more pain to the others who cared and loved you. Once a child dies he/she might forget they existed in this world but, the parents of the children will be devastated by the pain that caused them for the rest of their lives and will remember them forever. Gandhi’s death was one of the heartbreaking histories in mankind. His work has been never forgotten, people still remember the pain he suffered to achieve the goal that he aimed for. Hamlets death not only caused the pain to his best friend but also changed the Fortinbras point of view on the war. He realized that not only there are soldiers in war but, also they are in everyday life and that they try anything and everything to help others or to succeed in their own goal. In hamlet the character Ophelia, she kills herself by drowning underwater. Ophelila’s suicide devastated her brothers heart filled with so much sorrow and anger. Her death not only influenced his brother but also Hamlets mother Gertude because she wanted Ophelia to be her daughter in law as Gertrude says “I hop’d thou shouldst have been my Hamlet’s wife” (5.1.233). Malcom X use to say “Just because the heartbeat stop that doesn’t mean that all memories and feelings for the dead would go away”. Gandhi’s significance on sacrifice is “The sacrifice which causes sorrow to the doer of the sacrifice is no sacrifice. Real sacrifice lightens the mind of the doer and gives him a sense of peace and joy. The Buddha gave up the pleasures of life because they had become painful to him.” He said a sacrifice cannot be ended in sadness but it can be changed if the person receives peace from it. Innocent people died in Hamlet to give peace to King Hamlets ghost and to hamlet, so that he can make peace with his revenge. At the end of the play Hamlet knew he is going to die, this is obvious on several occasions where Hamlet illustrates to others that he just wanted to end his life of the treacheries. Sure, Hamlet’s actions throughout the play make him seem mad, but in reality, this madness was just a tactic of his in his plan to get revenge for his fathers’ wrongful death and justice. Contrary to accepted belief, the tragedy connected with Hamlet is not just about Hamlet or his own family. It is, however, about the sacrifices that the Polonius’ family has gave, whose deaths are not the result of any sins they omit but by their mortal manipulated by Hamlet and Claudius for explanations they are unaware of. Although the death of Polonius’ family stands out as being for the most part tragic, many other characters in the play are killed as well. In fact, the death of a character in Hamlet almost becomes commonplace near the end of the play. Perhaps Hamlet had never been taught killing out of anger was wrong, maybe he thought it was the punishment his Uncle deserved. Maybe the most important thing though is to be at peace with yourself, be happy with yourself, and with your actions. This above all: to thine own self be true (1.3.84), that is all that you can hope for from Hamlet; that he was above all else, true to himself.
UV Light and Cisplatin Induced DNA Damage Response. Simiao Lyu Abstract One of the essential function of an organism is to maintain its genome integrity, in order to survive and pass on its genetic materials to the next generation. However, as the carrier of genomic information, DNA is constantly being damaged by exogenous and endogenous factors. Therefore, through evolution, a variety of mechanisms that counter the effect of DNA damage emerged. In this study, we applied different dosages of UV light and Cisplatin, as DNA-damaging agents to SV40 T antigen immortalized MRC 5 Cells, to examine the various cell responses by assaying the relative concentration of key cell cycle regulatory proteins. Introduction In human body, DNA cells carried are constantly under assaults by exogenous and endogenous agents, causing a significant amount of lesions per day (Lindahl et al., 2000). If those lesions are not treated efficiently and the accumulation of lesions continues, mutations and aberrations of genome may lead to the emergence of many forms of physiological disorders, such as tumorigenesis, Parkinson’s disease and Friedrich’s ataxia (Stratton et al.,2009; Mirkin et al., 2007; Yang et al., 2008). One of the universal exogenous DNA damaging agent is ultraviolet(UV) light. UV-A and UV-B are its residues that are not absorbed by the ozone layer, and these residues may induce approximately 104 lesions to an exposed cell per hour (Doll et al., 1981). UV light is both a mutagen and potent cytotoxic agent ,which can trigger cell apoptosis by either accumulating DNA lesions or trigger CD95/Fas receptor and induce apoptosis directly (Rehemtulla et al., 1997). Another well-characterized DNA damaging agent is cisplatin. Unlike UV light, It is usually introduced to the human body during chemotherapy. (Prestayko et al., 1979) Researchers had already identified its DNA damage mediated apoptotic signaling pathway, which involves the TAB1 regulation of proteins’ circuitry (Yan et al., 2013). While DNA is damaged within a cell, there are many possible cellular responses, such as RNA processing, cell arrest and repair. DNA damage repair mechanisms varies according to the type of DNA damage. Apoptosis can be induced by accumulative DNA lesions to ensure the integrity of genome (Stephen et al., 2009) . Materials and Methods (Instructed By Practical Handout) SV40 T antigen immortalised MRC5 cell cultures SV40 T antigen immortalised MRC5 cells were split into eight experimental dishes with the appropriate medium. Those dishes were then incubated (at 5% CO2, 37oC) for two days and ready for cell damage and protein assay. Cell damage UV light and Cisplatin were employed as DNA damaging agents. For UV light, we chose 4 levels of dosages:64 J/m2 , 125 J/m2 , 250 J/m2 and 500J/m2; as for cisplatin, we choose to pipette 6.25uM, 12.5uM and 25uM to cells. All cells are damaged for 22 hours. Cell harvesting and protein assay After 22h, cells were harvested and prepared for protein assay. Immunohistochemistry and fluorescence microscopy were employed to indicate the level of proliferating cell nuclear antigen (PCNA), and western blotting technique were used to indicate the level of cyclin D1, geminin, tubulin and γH2AX for eight cell cultures separately. Hypothesis, Mechanisms, and Expected Results The hypothesis being tested were: For a cell culture, the overall type and magnitude of DNA damage response depends solely on the level of DNA damage they receive. According to our theory, while cells receive endurable damage, DNA repair mechanism will be favored statistically, which results in cellular arrest and DNA repair. Whereas, when the DNA damage level exceed a threshold value, cells will tend to proceed to apoptotic pathway. Since we assume there can only be two types of cell fates: cells suffered DNA lesions will either activate the arrest/repair pathway or they will activate the apoptosis pathway. Therefore, by protein analysis on certain key cell cycle regulatory proteins that are involved in those two pathways, we can see the relative level of protein expression and hence deduce the cell responses at different levels of cell damage. Here we choose to radiate UV light and pipette cisplatin at various dosages to different cell cultures. For UV light we choose 64 J/m2, 125 J/m2 as low dosages and 250 J/m2 and 500J/m2 as high dosages. As for cisplatin, we choose 6.25uM as low dosage, and 12.5uM and 25uM as high dosages. During this experiment, we examined the level of four proteins via immunoblotting: the first protein indicator we examined was cyclin D1. It is a protein that can be activated in the arrest and repair pathway. Previous investigations also suggest it will prevent low dosage UV-induced apoptosis and promotes adaptive resistance (Ahmed et al., 2008). The second protein we assayed was geminin. It was initially identified as an inhibitor of DNA replication (McGarry and Kirschner, 1998). Later investigations identified an increase in geminin level are correlated with increased proliferation of tumor cells, suggested that geminin may play a role in tumorigenesis (Wohlschlegel et al., 2002, Montannari et al., 2005). It seems contradictory though, overexpression of geminin in the Drosophila embryos leads to ectopic neural differentiation and cell apoptosis (Quinn et al., 2001). We believed, the functional role of geminin varies according to their presence in the different phases of the cell cycle. Under normal conditions, the majority of geminin will concentrate in G1 phase to regulate cell proliferation efficiently. Whereas, under pathological conditions, geminin will tend to present in S or G2 phase, where it has little influence on the regulation of cell proliferation.(Shreeram et al., 2002, Yoshida et al., 2004). The third protein we examined was tubulin. It plays a vital role during mitosis, and it is responsible for the production of spindle fibers in the metaphase (Wang et al., 2014). In this case, tubulin is an indicator of cell proliferation and apoptosis levels. The last protein we examined via western blotting was γH2AX, which is a sensitive molecular indicator for DNA damage repair. Because, when a double strand break is detected, one of the early cellular response will be the phosphorylation of H2AX, leads to the formation of γH2AX (Mah et al., 2010) . We also employed immunofluorescence technique to examine the level of PCNA in cells suffered DNA damages. Previous studies indicated that while UV light is inducing DNA lesions, p21 will be degraded, induce PCNA activation which then leads to DNA repair. (Soria et al., 2006) According to the functional mechanisms of different proteins, we would expect to see a relatively higher expression level of geminin,γH2AX, cyclin D1 and PCNA while a cell culture is damaged with low dosages of genotoxic agent, and vice versa. Because theoretically, these proteins are involved in cellular arrest and DNA damage repair responses. We would also expect the level of tubulin to decrease gradually while cell damaging level is increasing. Because DNA damaging agents can induce apoptosis and decrease the tendency of cell proliferation. Results The actual outcome of this experiment was astonishing. On the one hand, we acquired a relatively consistent green immunofluorescence assay result (Fig 1). The control dish shows a constant level of PCNA expression. When a low dosage of cisplatin was applied to the cell culture, the expression of PCNA tends to increase (Fig 1b), which suggested more cells were undergoing DNA repair. So far our experimental results are consistent with previous researches (Stelter and Ulrich, 2003; Soria et al., 2006). But, when 12.5uM of cisplatin were applied to the cell culture, the level of immunofluorescence drops dramatically. This inconsistency is likely due to human error during cell harvesting and preparation. Because microscopy indicates there were an unusual amount of crystals forming within 12.5uM of cisplatin slide, suggested cells may be left dried for an extended period of time, and proteins might be denatured. Interestingly, we expected cells to switch entirely to apoptotic pathways when we applied the maximum dosages of cisplatin to a cell culture (25 uM). However, we observed a significantly higher level of green immunofluorescence in Fig 1d, suggested its DNA repair pathway are the most active among all cisplatin treatments. Cell cultures that were treated with UV light had a similar trend in response: when cells were exposed to 63 J/m2 of UV light, the level of PCNA expression are slightly lower than that of the control dish. As we increase the intensity of UV light to 125 J/m2 and 250 J/m2, the level of PCNA decreases accordingly, indicates that most of the cellular responses had switched to the apoptotic pathway. However, the result of the maximum dosages treatment (500 J/m2) were out of our expectation: like the maximum dosages cisplatin treatment, cells which endures maximum dosages of UV light exposure turn out to have the highest level of PCNA expression. On the other hand, the result from immunoblotting is not consistent enough to be analyzed. By cross-comparison with other groups, we found out that no group observed any band from tubulin analysis, and there are heavy non-specific binding occurs on all cyclin D1 analysis. These outcomes indicated inappropriate secondary antibodies were selected for CD1/tubulin assay. However, as for γH2AX and geminin assay, we were misguided by the vague molecular marker; hence we cut on the incorrect side, therefore, there are only a few bands on the edge (fig d). Discussion Previous investigations indicated: there are multiple cellular pathways which enables cells to perform DNA damage repair (Teruaki and David, 2013). Such a response is completed by an orchestration of protein activities, including some of the proteins we analyzed during this experiment, such as PCNA. Some previous studies stated that an increasing level of DNA damage leads to a rising PCNA expression (Balajee et al., 2001). These observations partially agrees with our experimental results. However, by applying a relatively higher dosage of damaging agents, we also realized that, after passing through a certain “threshold”, the rise of genotoxicity could results in a gradual decrease in the level of PCNA, which we thought was due to a decrease in the amount of cells undergoes DNA repair and more cells undergoes apoptosis. This observation was also consistent with past findings, which indicates that PCNA activities can be maximally triggered under low levels of UV radiation(Soria et al., 2006). However, none of any published reports observed sudden increase of PCNA expression when the level of DNA damage was raised from high levels to maximum dosages. There can be three possible explanations for such observations: the first and most likely explanation is: under low dosages, when DNA were repaired the expression of PCNA drop down to base level, whereas under maximum damage, cellular repair pathways are activated throughout the damaging process. The second possibility is: we made errors during experiment and cell cultures were stained differently. Last but not least, there might be a secondary threshold of DNA damage level that leads to acute cell cycle arrest followed by DNA repairing instead of cell apoptosis. Further studies needs to be conducted to testify our expected results on cyclin D1, geminin, tubulin and γH2AX. Possible Improvements To improve experimental design, we could devise more appropriate UV dosages to maintain the consistency of the experimental outcomes, because the result showed us UV light possess higher genotoxicity compared to cisplatin. We should also consider cell fates other than repair and apoptosis to post a more realistic hypothesis. Finally, a separate experiment to analyze the effect of time length of cell damaging to the cell responses, can be conducted to backup our theory. As for experimental procedures, besides avoiding possible human errors, our experimental results could be more convincing if we were able to implement following improvements: firstly, we could split cells into at least 16 dishes. This improvement provides larger sample size and testing capacity. Secondly, microscopes with high magnification could be used to perform cell counting to analyze the level of cell apoptosis and cell arrest quantitatively, instead of speculating based on the level of indicators. Last but not least, a quantitative measurement for protein concentrations, such as UV absorption spectrum and iTRAQ technique, could be applied to produce a more reliable result. Conclusion Nowadays, we have made much progress toward the understanding of DNA damage responses. However, Some detailed mechanisms, for example, the regulation of protein activities in DDR and the exact magnitude of cell responses toward DNA damage, still remains mysterious to us. Although our experiment failed to unveil this mystery, many published experimental data do support our theories to some extent. Further investigations should be conducted to fully understand the orchestration of DDR mechanisms, and such knowledge will enable us to develop more effective clinical applications to treat DNA damage induced diseases. UV Light and Cisplatin Induced DNA Damage Response
Human Resource Management – Linking Strategy to Practice
Human Resource Management – Linking Strategy to Practice.
This assignment consists of two (2) sections: a narrative and a PowerPoint presentation. You must submit two (2) sections for the completion of this assignment. Label each file name according to the section of the assignment it is written for.Note: For additional information on how to submit more than one file for an assignment, follow the instructions in the document “How to Submit Multiple Files for an Assignment”, located here.Imagine that you have just been hired by a new company as the director of the HR department. You have been tasked to hire a new secretary for the department and to develop an employee compensation and benefits package that will be used for that position upon hire. Develop a PowerPoint presentation to present this information to your Vice President. Go to the Bureau of Labor Statistics’ (BLS) Website, located at www.bls.gov, for information regarding organizations and pay in your geographical area.
Human Resource Management – Linking Strategy to Practice
Benneton’s group
online assignment help Benneton’s group.
– Includes a general understanding of how the problem applies, analysis based onindustry and internal positioning, recommendations etc- I need 2 double spaced pages.https://centridiricerca.unicatt.it/cersi-wp12009.PDFww.benetton.com – use these two sites for better information or if you find any better information from outside use that but only academic sources.-I only want you talk about question #2 only about ORDER QUALIFIERS not on order winners. please keep that in mind. – Talk only about the Operational activities of the Benneton’s group such as production, manufacturing, Supply chain and so on which are the daily activites and less then 6 months of period. It is your job to find the operatinal activities of the company and talk about them. Also, talk about their gloabal sales revenue. basically how much they sale.- I want you to talk specifically about the answer no intro and no conclusion. I need really good answer if you can do it then take it please don’t take it. I expect so much in the answer.
Benneton’s group
HSPM 108 UCSD Factors to Consider when Implementing the Technologies Discussion
HSPM 108 UCSD Factors to Consider when Implementing the Technologies Discussion.
You are the General Manager of a 500-room hotel. Your hotel also has kitchens, restaurants and meeting rooms. Answer the following two questions:1. Select 10 technologies from all of the information and communication technologies that we have learned in this course. Discuss how you could utilize them to operate effectively and efficiently? Why?2. What are the key considerations in selecting and implementing these technologies (systems)? Hint: Think about costs, service delivery, operational issues, staffing and training, management needs, and so on. Requirements:Save your file as lastname_firstname_108_final, 10% penalty for not following this format.800 words total (+/-10% is OK). Include a word count at the end of your essay. Word count should not include references.Grading will be based on the following rubric.Single space with proper margins. You should properly cite and reference the research and information used. At least five references are required (APA style preferred), which should not be included in word count.Save your file in MS Word or PDF format. Canvas will accept files only in doc, docx, and pdf format.Submit your write up to HSPM 108 Final Exam Part 2 on CanvasDue: 5/19/2020 at 11:59 pm.Important note:Your write up will be reviewed by Turnitin. Without properly citing and referencing the materials used, you may get a high Turnitin Similarity Index, i.e. above 20%. In such a case, you should review your paper and make sure you do not violate University’s policy on plagiarism.
HSPM 108 UCSD Factors to Consider when Implementing the Technologies Discussion
Gender Studies homework help
Gender Studies homework help. Final PaperPlease read these assignment instructions before writing your paper, and re-read them often during and after the writing process to make sure that you are fulfilling all of the instructions. Please also utilize the assignment guidance and the outlined model provided.Overview In the Week One Assignment, you formulated a concrete ethical question, took a position on that topic, and identified a reason supporting and a reason opposing that position. In the Week Three Assignment, you discussed either deontological or utilitarian theory, applied that theory to the question, and raised a relevant objection.By engaging with the course material, you now have had a chance to refine your thinking and broaden your understanding of the problem by approaching it from the perspective of multiple ethical theories.In this paper, you will demonstrate what you have learned by writing an essay in which youPresent a revised formulation of the ethical question and introduction to the topic.Explain the kind of reasoning you think is the best way to approach this question, and how that reasoning supports the position you think is strongest.Raise an objection, and be able to respond to it.Instructions Write an essay that conforms to the requirements below. The paper must be 1500 to 2000 words in length (excluding the title and reference pages) and formatted according to APA style as outlined in the Ashford Writing Center.The paragraphs of your essay should conform to the following guidelines:Introduction Your first paragraph should begin with the topic question, suitably revised. It should be focused, concrete, and on a relevant moral problem. You should then introduce the topic in the way described by the Week One instructions, but reflecting the developed understanding and information you have gained about the topic and any necessary refinement of the scope. Follow this with a thesis statement that states your position, and a brief description of the primary reason(s) supporting your position. (See the handout on thesis statements provided). Finally, provide a brief preview of the overall aim and procedure of your paper.Explanation and Demonstration of Moral Reasoning This section of the Final Paper will explain and demonstrate what you believe to be the best way of reasoning about the question you have chosen, and showing how that reasoning supports the position you have taken on the question. You might explain the principles, rules, values, virtues, conceptions of purposes and ends, and other general ideas that you find persuasive, and show how they support concrete judgments. In the course of doing so, you must make reference to at least two of the approaches that we have examined in the course (such as deontological, utilitarian, or virtue-based), and utilize at least one resource off the provided list for each of the two approaches. One of these theories may be the theory you discussed in your Week Three Assignment, but your discussion here should be more refined. For example, you might find the reasoning associated with Aristotelian virtue ethics to be the most compelling, and reference Aristotle in the process of showing how that reasoning supports a certain conclusion. In the course of this, you could contrast that with a utilitarian approach, referencing Mill for instance.Objection and Response After explaining the ethical reasoning that supports your position, you should raise an objection and respond to it. An objection articulates a plausible reason why someone might find the argument weak or problematic. You should explain how it brings out this weakness, and do so in a way that would be acceptable to someone who disagrees with your own argument. Then, provide the best response you can to the objection, showing how it does not undermine your position. Your response should not simply restate your original position or argument, but should say something new in support of it.Conclusion Provide a conclusion that sums up what you presented in the paper and offers some final reflections.Resource Requirement You must use at least four scholarly resources. Two of the resources must be drawn from the list of acceptable primary resources on each of the two theories you discuss. For example, if you discuss deontology and virtue ethics, you would need at least one resource under the ?Deontology? list and at least one resource under the ?Virtue Ethics? list. The other two may be from either the Required or Recommended Resources, or scholarly resources found in the Ashford University Library.The textbook may be cited, but it does not count toward the resource requirement. If you cite the textbook, you will still need to cite at least four more sources that fulfill the requirements stated above.If you need help with finding additional resources, or are unsure about whether a particular resource will count toward the requirement, please contact your instructor.For sources to count toward the resources requirement, they must be cited within the text of your paper and on the reference page. Sources that are listed on the references page, but not cited within the paper, do not count toward fulfilling the resources requirement.For information regarding APA, including samples and tutorials, visit the Ashford Writing Center.The Final Paper:Must be 1500 to 2000 words in length (excluding title and reference pages), and formatted according to APA style as outlined in the Ashford Writing Center.Must include a title page with the following:Title of paperStudent?s nameCourse name and numberInstructor?s nameDate submittedMust begin with an introductory paragraph that has a succinct thesis statement and statement of procedure.Must address the topic of the paper with critical thought.Must end with a conclusion that reaffirms your thesis.Must make meaningful reference to at least two of the ethical theories studied in the course.Must use at least four scholarly resources that fulfill the stated requirements.Must document all sources in APA style, as outlined in the Ashford Writing Center.Must include a separate reference page, formatted according to APA style as outlined in the Ashford Writing Center.Carefully review the Grading Rubric for the criteria that will be used to evaluate your assignment.Gender Studies homework help