What happened after an agreement upon slavery and the expansion could not be rea.
A.The southern states began to consider succession.B. The government decided to go to war with Mexico.C. The nation took steps towards electing another president.D. The government took measures to revise the US Constitution.
Mitochondrial DNA Analysis: Techniques and Applications. Introduction Because of advancements in technology and techniques, mitochondrial DNA (mtDNA) analysis has become a common method in forensic procedure. Polymorphisms in human mtDNA were first discovered in 1980 (Brown 1980) while a complete sequence was first achieved in 1981 (Anderson et al., 1981). MtDNA sequencing is frequently used in cases where biological samples are degraded or low in quality, instead of nuclear DNA (nDNA), because each cell contains more than 1000 copies of mtDNA per cell instead of only two per cell in the case of nDNA. This article will examine the background information and techniques of mtDNA analysis as well as several of its applications. Background Information Mitochondrial DNA is located outside the nucleus of a cell in organelles known as mitochondria. The chief advantage of mtDNA is the fact that it is present at a rate of between 100 and several thousand times per cell, making it much easier to be typed than nuclear DNA (James and Nordby, 2005). This characteristic makes mtDNA a very useful tool in a sample that is either degraded or limited in quantity. Its typical sources include hair, bones, teeth and bodily fluid such as saliva, blood, and semen. Another feature of mtDNA is that it is maternally inherited so a sample can be collected from any member of the maternal lineage (Giles et al. 1980). The mtDNA for humans is ~16,569 base pairs and consists of two regions, the control region and the coding region. The control region is located in the displacement loop (D-loop), and contains hypervariable regions 1, 2, and 3 (HVI, HVII, and HVIII). HVI and HVII are sufficiently polymorphic to allow two samples to be differentiated. However, HVIII is currently being assessed to determine its capacity for use in discriminating between individuals and could one day be used to increase the discerning power of mtDNA. The D-loop of mtDNA does not contain any functional genes, and accumulates mutations at a rate of up to 10 times that of nDNA (James and Nordby, 2005). See image below for a sample mtDNA structure. The HVI region is approximately 341 base pairs (bp) in length while the HVII region is 267bp. The other regions in the mtDNA genome have been successfully analyzed but have not been frequently used in forensic works because they haven’t been proven sufficiently hypervariable. Techniques of mtDNA Analysis The methods for mtDNA typing have gradually changed over the past thirty years. The process has gradually changed protocols from low-resolution restriction length polymorphism (RFLP) analysis to sequence analysis of the HVII and HVII regions and is moving towards a complete sequence of the mtDNA genome for each case. The sequencing and comparison of the mtDNA can be broken down into four steps: Extraction, polymerase chain reaction (PCR) amplification, sequencing, and comparison. Extraction A biological sample contains main substances besides DNA. The purpose of extraction is to separate the other material from the DNA before it is examined. The sample is prepared and mixed with certain organic chemicals that lyse the cell membranes, separate proteins from the DNA, and then denature the proteins and destroy their structures to decrease their solubility. Using a phenol chloroform, the denatured proteins are removed. The DNA is then purified using ethanol precipitation and isolation by centrifuge. PCR Amplification The polymerase chain reaction is process similar to the one used by cells to copy their own DNA. By using this process, a relatively small number of copies could be multiplied into over a billion in around thirty cycles. PCR amplification is a three step process. First, the two mtDNA strands are denatured by heat at 94°C. This means the strands separate from each other into two equal and opposite strands. Second, the sample is cooled to 60°C and the primers bind to the DNA template, which gives a start sequence to the DNA polymerase in step three, which extends the primers by adding the respective nucleic acids to the base strand and completes the sequence, thus turning one copy of the DNA into two. By repeating this process, the number of copies increases exponentially. Sequencing The primary sequencing method in mtDNA analysis is known as the Sanger method, which is a six step process that builds off the PCR amplification. First, the removal of remaining Deoxyribonucleotide triphosphates (dNTP) and primers from PCR through spin filtration or enzyme digestion is required. The PCR quantity is then found to determine if there is enough product to sequence the DNA. Then, four different colored fluorescent dyes are attached to the four differenet ddNTPs. After that, the unincorporated dyes from the sequencing reaction are removed using spin filtration. The purified sequencing reaction is then diluted in foramide and separated through gel electrophoresis. Finally, the sequence analysis of each reaction performed is compiled and interpreted. Comparison After sequencing the HVI and HVII regions of the DNA, the sequences are confirmed with the forward and reverse strands for that sample. Differences are then noted from the revised Anderson sequence, which is the newest standard for mtDNA. Finally, the known and the unknown samples are compared with each other and then compared with the database to determine halotype frequency. Applications As previously stated, current applications of mtDNA analysis are best suited to DNA which is very old and highly degraded. Some of the applications in this include individual identification, evolutionary biology, and maternal lineage testing. Individual Identification MtDNA can play a considerable role in the identification of individuals in which the biological sample would be highly degraded, for example, identifying remains of unknown soldiers. In June, 1998, a DNA sample from one of the US tombs of the unknown was collected and compared to reference samples from seven different families who had lost a family member during the war. The results eliminated six of the families, and provided a positive identification on the final family. The results proved consistent and the soldier was identified as Air Force First Lieutenant Michael J. Blassie. In this case, the small, closed population allowed relative ease of identification. To date, the remains of around 150 soldiers from the Vietnam War and 1,200 from the Korean War have been reunited with their families. Evolutionary Biology There are two main hypotheses about human evolution and phylogenic trees trying to guess where the human race branches off from the rest of the animal kingdom and spread all over the world. The multiregional evolution suggests that modern humans evolved from Neanderthals and Homo erectus at the same time in different parts of the world. This hypothesis is supported by fossil evidence, particularly a gradual change in facial structure from earlier to modern humans. The other hypothesis suggests a more recent African origin around 100,000 – 200,000 years ago, in which a small group of modern humans populated the rest of the world. This would have been done without mixing genetic material with other forms of humans. Because the human mitochondrial genome was one of the first to be completely sequenced, it took researchers a while to see the advantage of sequencing the entire genome. First, mutations in the D-loop occur at a rate 5-10 times faster than that of normal nDNA, and having the complete genome showed the same polymorphism on two different loci. Also, while the D-loop was changing at a high rate, mutations outside the D-loop were near zero, allowing the rate of evolution for the rest of the genome to be seen clearly and evenly between different complexes. The data collected gives evidence to support the recent African origin theory. By finding the substitution rate between the sequences, it is possible to find dates at which the genetic material coincided with itself and thus makes it possible to find approximate dates and points on the phylogenic tree. From this information, the sequence of events in human evolution can be established. The data suggests a severe population constriction around 180,000 years ago. (Brown, 1980) This evidence supports the recent African origin hypothesis while with the multiregional hypothesis you would expect to see this constriction at an older date. Maternal Lineage Because mtDNA testing can be done with a smaller quantity of DNA, it is well suited for maternity testing and forming a family tree. Right now, mtDNA is the most common form of DNA analysis performed to determine parental maternity testing today. Obviously, this is used in the legal system for the purpose of deciding custody battles. MtDNA also has precedence in accurately performing historical identifications through maternal lineage. A famous case of this would most likely be the case in which Jesse James’ body was identified by using a comparison between his DNA and the great-great-granddaughter of his sister. This lineage would provide a positive match if in fact it was Jesse James’ remains, which in fact it was. However, the James case was not without its faults. The initial attempts at mtDNA analysis of the bone specimens from the specimen were not successful. More, specifically, there was no product formation during the Polymerase Chain Reaction. The most likely cause of this type of error would be the poor condition the material was in.(Stone et al. 2001) After running several tests on the bones, teeth and hair of the remains, mtDNA was collected and analyzed from two molar teeth and two hair fibers. This just shows that while mtDNA analysis has the possibility for up to 100% exclusion, because of mix-ups, laboratory errors, contamination, and degradation, it’s not 100% accurate. Conclusion Because of its impact on the forensic community, mtDNA analysis has become a power discrimination tool in forensic science. The methods that have been developed over the past thirty years have been firmly founded in scientific research. Each tool used in the aid of sequencing the mtDNA has a specific purpose that plays an important part the analysis as a whole. As technologies evolve, the discriminating power goes up, as in the case of adding the HVIII loci to the standard, and the error rates go down because of new information regarding contamination prevention procedures and determining the presence of degraded sample. Finally, by applying all of these concepts to the forensic world, mtDNA analysis becomes a powerful tool that has the potential to trace lineages, prove maternity, and identify unknown persons. References Anderson, S., Bankier, A.T., Barrel, B.G., Bruijn, M.H.L., Courlson, A.R., Drouin, J., Eperon,I.C., Nierlich, D.P., Roe, B.A., Sanger, F., Schreier, P.H., Smith, A., Staden, R., and Young, I.G., 1981, Sequence and Organization of the Human Mitochondrial Genome.MRC Laborator of Molecular Biology, Hills Road, Cambridge, UK Brown, W. M. 1980 Polymorphism in mitochondrial DNA of humans as revealed by restriction endonuclease analysis Proc. Nati. Acad. Sci. USAVol. 77, No. 6, pp. 3605-3609 Butler, J. 2005, Forensic DNA Typing. Elsevier Science pp.242-298 Giles, R.E., Blanc, H., Cann, H.M., and Wallace, D.C. .1980, Maternal inheritance of human mitochondrial DNA, Proc. Nati. Acad. Sci. USAVol. 77, No. 11, pp. 6715-6719, Ingman, M. ,2001,Mitochondrial DNA Clarifies Human Evolution, http://www.actionbioscience.org/evolution/ingman.html Image Source: MtDNA. http://www.nfstc.org/pdi/Subject09/images/pdi_s09_m02_01_a.1_large.jpg James, S.H. and Nordby, J.J. 2005, Forensic Science: an introduction to scientific and investigative techniques. N.W. Ciroirate Blvd., Boca Raton, Florida Michael Blassie Unknown No More, 2006, http://www.nlm.nih.gov/visibleproofs//galleries/cases/blassie.html Stone A.C., Starrs J.E., Stoneking, M. Mitochondrial DNA analysis of the presumptive remains of Jesse James. J of Forensic Sci 2001;46(1):173-176. Mitochondrial DNA Analysis: Techniques and Applications
Final Paper – Advanced supply Chain Management.
What happened after an agreement upon slavery and the expansion could not be rea
Describe both the business strategy and supply chain strategy for your organization, or for a business unit of your organization, or for some other organization for which you have worked in the past and still retain contacts. Your research paper will describe how this organization aligns its business strategy with its supply chain strategy (use the definition from page 26 of the text to guide you) if they have a formal one. You are to expand this description and supplement it with a critical analysis and suggested improvement steps/actions based on what you have learned in this course.Key issues to address:What opportunities exist to improve supply chain activities through
better alignment of logistics and the company’s strategic plan?In what ways can the supply chain integrate with logics in order to get goods and services moved effectively and efficiently?How might a company’s agility in terms of the supply chain and/or logistics improve the movement of goods and /or products?What challenges does the interaction of supplier and manufacturers bring to the logistics management arena?What processes could be adopted to increase value for the customer?What are the impacts of Supply Chain Management on various organizational functions?What are the financial implications of improving supply chain management for the company?What “best practices” are available to support your recommendations? (10-15 pages) (6-8 references)
Final Paper – Advanced supply Chain Management
Robert Lowell’s The Drinker | Analysis
Almost 21 million Americans have at least one addiction, yet only 10% of them get treatment (Addiction Center). For Robert Lowell, poetry served as his treatment. In 1917, Lowell was born unplanned and unwanted. His life did not follow a continuous line: it was convoluted like razor wire through intervals of misery, each a discouraging echo of the last, a toilsome way to exist in time. Imagine living life in a strait jacket inside a white padded room, surrounded by the deafening silence that is depression. Lowell’s grappling fight with his manic depression and substance abuse led to his confessional poem, “The Drinker,” in which he directly criticizes what one can assume is himself. In “The Drinker” Robert Lowell uses alliteration, metaphors, and imagery to posit that when a person suffers from substance abuse, they often become victims of a brutal cycle until they ultimately fade into isolation. At the beginning of the poem, Lowell uses alliteration to exemplify the dangerously repetitive and abusive cycle of alcoholism. In the second stanza, the persona recounts his daily routine of being “stubbed before- breakfast cigarettes”(5). The man reflects on the status of his life. The alliteration of “stubbed before-breakfast,” may emphasize how the man had slowly faded into seclusion without his significant other, by slowing down the rhythm of the poem. The memories of her had burned with the smoke of his “breakfast cigarettes.” As the stanza continues, the persona describes the state of his nightstand. He notices that the cigarettes, “burn bull’s eyes on the bedside table” (6). The alliteration of “bull’s eyes…bedside” offers another level of complexity that may suggest that his repetitive binge drinking caused his partner to leave him. His nightstand that perhaps once held his partner’s things, was tainted by “bull’s eyes” from his cigarettes. What once housed their possessions now held his alcohol. In the seventh stanza, the persona describes the objects that his significant other left behind that constantly reminded him of his isolation. As he describes these objects, he confesses that, “her absence hisses like steam” (24). The alliteration of “hisses…steam” mimics the hissing of his partner’s absence. The hissing stayed with the man despite her absence; he still felt a connection with this woman and regretted his loss. Lowell uses alliteration to relate his substance abuse to his isolation and loss of his potential wife. Throughout the poem, Lowell employs metaphors to evince the idea that redemption is often unattainable. The first line begins with the statement, “The man is killing time.” (1). While a common phrase, in this context, it suggests a lot more. The man can actually be considered as “killing” time, in that drunkenness allows time to disappear. He suggests that drinking is not just something the man does for fun or out of habit. Instead, he wants to kill time. He wants to escape from his life. As the poem continues the persona describes, “leagues of ocean, gasping whiteness, the barbed hooks fester” (11-12). The man is drowning in “leagues of oceans” without his significant other to help pull him out of the menacing cycle, that became his addiction. He was unable to fight it, leaving him disoriented and in “gasping whiteness.” The “barbed roots” of his addiction were too deep-rooted to pull them out. The man is stuck in a life where he doesn’t have the will to pull the hooks, or the alcohol, which had been his lifeline for so long, out. In the eighth stanza, Lowell adopts an allusion in an attempt to explain his struggle. He posits that “No voice outsings the serpent’s flawed, euphoric hiss” (30-31). In the context of the stanza, the man attempts seeking redemption however, the drugs have overtaken him. Nothing he wants could “outsing” his need for and dependence on the drugs. His biblical allusion insinuates that his addiction has become a rite of passage, in that he is doomed to become isolated. Lowell’s manifestation of metaphors throughout the poem, hinted at his failure to escape his addiction. As the poem continues, Lowell administers imagery to demonstrate how addiction often results in isolation. As the man looks for his neighbors, “their names blur in the window” (13-14). The “blurr[ing] window” suggests the man’s fogged view of the outside world. His clear view becomes tainted by the smoke of his cigarettes. The man’s figurative window closes, he can only see what could have been, through its transparent surface. He can’t see past his regret. Later in the poem, the persona describes the condition of the room he’s in and notices that “the cheese wilts in the rat-trap” (32). Without his partner, his well-being as well as his state of living “wilt[ed],” like the rat trap. His entire quality of life diminished without the help of the woman. Finally, the poem’s last stanza begins with, “out on the street two cops on horseback clop through the April rain” (37-38). The line “out on the street,” implies the man’s confinement to the wall in front of him from the outside. He’s been rejected from the world. The “April rain,” provides an image of the man standing in front of the water-speckled window. The window shapes the way he thinks and distorts all on the other side. Lowell employ’s imagery to impactfully illustrate the extent of his isolation. In “The Drinker,” Lowell demonstrates how a person’s history and habits can adversely affect their future. Sometimes depression can make a person feel isolated. Lowell’s application of a third-person point of view describes his feeling of watching himself from the outside, an almost out of body experience. He is an unwilling witness of his descent into seclusion, “wasting time”, killing himself. Throughout his life, Lowell became further and further imprisoned in the inescapable clouds of depression and addiction. The waves of depression, pummeling his brain as his body “founders” deeper into the inky, helpless, cold depths of the ocean floor. The question: would he stay trapped, or fight to the surface and live? Ultimately, the poem concludes with the feeling that redemption is impossible. So, the ocean hisses with the echoes of the souls trapped by the perilous cycle of depression. “Addiction Statistics – Facts on Drug and Alcohol Use – Addiction Center.” AddictionCenter, www.addictioncenter.com/addiction/addiction-statistics/.
Argumentative essay assignment
assignment helper Argumentative essay assignment.
4/6 pages MLA formatting I have already began working on into and thesis However it needs to be condensed in a way that it still answers the essay promt please try not to change what I have written unless it’s in a way that makes the wording flow easier I don’t like when I’m trying to get help and the tutor doesn’t even use what I have done Or explained why it was incorrect I have an outline and enough work to put an essay together I would like someone that is willing to help me so that we are working on it together in a way This isn’t due until Sunday but I still need you to please consistently upload so that I may give you feedback And same for me
Argumentative essay assignment
EDUC 6166 Walden University Leadership Styles and Behaviors Essay
EDUC 6166 Walden University Leadership Styles and Behaviors Essay.
Assignment: Course ReflectionTo prepare for this assignment:Reflect on what you have learned throughout this course. Then, write a 2- to 4-page reflective essay that includes the following parts:Briefly describe the three most important concepts you learned in this course that will guide you as a director of an early childhood program. The concepts or strategies that you choose can be broad or narrow in scope. Explain how you will apply these concepts to your position as a director of an early childhood program.Based on one of the concepts or strategies that you identified, describe two actions that you will take within the next month (or when you begin your tenure as a program director) to pursue your efforts in this area. Provide a rationale for your selection.Reflect on what you had hoped to gain from this course. With these goals in mind, what further questions do you have regarding the topic of this course, and/or what additional information would you like regarding any of the topics presented? What steps will you take to obtain information that will answer your questions or to further explore the issues addressed in this course? Include the resources you have already discovered on your own and describe how they will help you in your position as a director of an early childhood program.
EDUC 6166 Walden University Leadership Styles and Behaviors Essay
Special Interest Disability and Personal Interview Qualitative Research Essay
Special Interest Disability and Personal Interview Qualitative Research Essay. Introduction Early diagnosis of chronic diseases is important in the prevention of disability. Health care professionals should try as much as they can to identify a disease early and start treatment before disability sets in. Besides, disability affects the psychological, social, spiritual and economic life of a person. This paper presents a report of the interview of a patient with fibromyalgia. It include the summary of the disease, description of the individual interviewed, results of the interview, application of the developmental theories and concepts as well as summary and conclusion. Brief summary of disability Fibromyalgia is a condition in which the patient has pain as well as tenderness in most parts of his body especially the joints and the muscular parts (Ostalecki, 2007). It is common in women who are between the ages of twenty five to fifty five. This condition is associated with general malaise, insomnia, frequent headache, stress, anxiety and depression. Health care professionals do not know the exact cause of fibromyalgia. ClauwSpecial Interest Disability and Personal Interview Qualitative Research Essay