Insulin and Erythropoietin Production

Insulin and Erythropoietin Production Insulin is a protein (polypeptide) discovered in 1921 by Banting with the pancreas being the site of its production. It is made up of 51 amino acids, divided into 2 chains; A and B, bonded by disulfide linkages. Chain A is made up of 21 amino acids with an intra-disulphide linkage, while chain B is made up of 30 amino acids (4). Why Insulin? Insulin is important in glucose metabolism, and is being used for the treatment of Diabetes mellitus; a metabolic disorder of glucose in the body. Initially, Insulin from animals was used to treat this disorder however nowadays synthesized human Insulin is being used, this is because; it is fast absorbed by the body, it has less allergic reactions, it contains less impurities, and it produces good results (3). Recombinant process of producing Insulin Synthetic Insulin was first produced in 1983 through genetic Engineering, which involve extraction of the human DNA (1), once extracted, the gene for Insulin is isolated, and enzymes are used to cut it. The gene is then cut using enzymes and put into the plasmid of a vector, where in most cases E. coli plasmid is used. Since Insulin contains two chains, two pieces of DNA are extracted, and the genes for the two chains are linked to ÃŽ ² galactosidase enzyme of the bacteria. The plasmids formed are then inserted into a host cell E. coli and sealed using another enzyme called ligase. And the host on replicating produces the enzymes each containing one of the two chains each. Production is followed by extracting and purifying the chains which are mixed in a reaction to reconstitute the disulphide bridges (1). ESCHERICHIA COLI AS RECOMBINANT INSULIN HOST Entero-bacillus, gram-negative E. coli is about 1 2ÃŽ ¼m, it can survive in the presence/absence of oxygen, and it also grows in an optimum pH and temperature of 7.0 and 37oC respectively. It utilizes glucose as its major carbon source and can also use other carbon sources like pyruvate, glycerol, acetate, and other sugars. K-12 and B strains are mostly used in the laboratory (20) Reasons for choosing E. coli Genetic Engineering technologies were developed using E. coli as a role organism, and so, the genetics of E. coli are well known among other microorganisms, as such its the most used organism for the production of different proteins (14). Moreover E. coli has a well known safety and production abilities, stable plasmid, controllable promoter, cheaper and easily cultured (6), E. coli also has fast growth rate, its easy to handle, and has well known fermentation skills and the ability to produce high protein content (14). That is why most of the proteins licensed recently by FDA and EMEA, were produced in E. coli (5). With these, and the fact that Insulin is a simple polypeptide (protein) which does not require glycosylation for its bioactivity and stability, E. coli carrying the plasmids for production of insulin will be used as the host for the production of Insulin Strain and plasmids: BL21 strain containing the pMYW-A and pMYW-B plasmids and temperature repressor ÃŽ »-c1857, will be used for insulin production (21). Growth strategy The various growth strategies that will be used to grow E. coli in order to make it happy and produce the desired product (11) include: Medium: E. coli needs nutrients like carbon, nitrogen and others; thus a carbon source; glycerol will be provided since its cheaper and more soluble than glucose (12), a source of nitrogen in the form of ammonium sulphate will also be provided. However such nutrients in large quantities can inhibit the growth of E. coli, as such a defined medium that contain optimum concentrations 20gl-1 glycerol and 2gl-1 ammonium sulphate will be used (11). The medium will also consist of the following; 3gl-1 KH2PO4, 1gl-1 MgSO4.7H2O, 0.8gl-1 citrate, and 6gl-1 K2HPO4 (23). Some trace elements will also be added to the medium. (23) Process and culture-strategies: E. coli will be grown submerged in a sterile controlled stirred tank reactor, and fed-batch will be used as the growth strategy so as to avoid accumulation of acetate which can be inhibits its growth, and reduce the production of the insulin (18). The growth strategy will be divided into two; initially batch mode will be used to initiate growth, after which the fed-batch exponential feeding will be used to produce the insulin (21). After adapting the medium and feeding method, oxygen transfer rates (OTRs) had to be increased through a suitable bioreactor design and over-head pressure (16). Large scale reactors usually reach high ORTs using air and normal aeration pressure, and so the oxygen partial pressure (pO2) will be increased by adding pure oxygen to the air-stream entering the reactor, thus increasing its oxygen transfer rates (16) DO will be maintained at 40% of air saturation and aeration rate at 1vvm. Foaming arising due to large number of cells and high aeration-rates will be solved by use of impellers for stirring simultaneously at 300rpm and the use of antifoam (ucolub N115) (16, 21). The process temperature and pH will be maintained at 30oC and 6.8 respectively so as to avoid partial proteolysis of the insulin protein. Bioreactor Design: Bioreactor vessel is usually cylindrical and made up of stainless steel. It is composed of impeller for stirring, Air sparger is placed at the bottom of the vessel for introduction of air, it has some inlets for introduction of acid/alkali for pH control and also for introduction of antifoams, nutrients and inoculum; It is also has pH, DO and temperature probes for sensing (22), Microbial activity during fermentation usually produces heat, so the bioreactor design must allow for removal of heat, and this can be achieved by cooling with jackets and coils (16) Bioreactors must also be designed in a way that it can withstand high temperature and pressure and to allow cleaning-up and sterilizing (22). Growth analysis Temperature, pH, DO, foam, partial oxygen and carbon dioxide pressures, will be analysed on-line, other parameters like biomass, will be analysed by using optical density (OD600) and dry cell weight (offline). Cell viability will be analysed by using flow cytometry, the concentrations of substrates and metabolites by enzymatic methods while insulin will be analysed using electrophoresis methods like SDS-PAGE, and ELISA, while its purity will be determined by HPLC (8). Limitations/Problems There are several problems that may arise during processing and can limit the use of this organism for Insulin production, these are; Poor secretion because of the structure of its membrane (and tough cell wall), small amount of foldases, chaperones and increased concentrations of proteases, leading to low productivity (7). Solutions to this problem include all measures taken to increase quality of secretion and production such as: Use of secretion systems like the system of ÃŽ ±-haemolysin (7) co-expression after co-cloning of foldases and chaperones (13) Improving the rates of gene-expression and using proteases deficient mutants like BL21 (18). use of E. coli mutants that are deficient of cell-wall (12) Limited post translational-modifications; including disulfide-linkage formation, which is important for the insulin stability and biological activity (9). Solutions to this problem include; Production of insulin with altered amino acid sequences through genetic engineering (9) Using E. coli mutants to enhance the formation of disulfide linkages e.g. Origami (15) iii. Exporting proteins into the periplasm which has disulphide bonding mechanisms (19). Codon biases; due to large quantities of exact transfer-RNAs found in E. coli, the codons in the human-genes are often different from those that are found in this organism. This results in inefficient expression of some of these rare codons by the organism resulting in an unexpected protein synthesis termination or wrong incorporation of the amino acids (12). This problem can be solved by replacing codons that are rare in the desired gene by codons that are often found in the E. coli and by co-expressing the rare transfer-RNAs (15). Acetate is usually formed as a by-product, and is inhibitory to growth of the cells (20). Solution is by using a fed-batch feeding method and by limiting DO level (11). Another problem is that large proteins are often obtained in an insoluble form (5); forming aggregates called inclusion bodies; IBs (20). This can be solved by adjustment of temperature, increasing the strength of the promoter, adjusting the number of plasmids, concentrations of the inducer, and the composition of the media (9). Erythropoietin EPO EPO is a glycoprotein that is produced in the renal cortex of the kidney (10, 11). It has also being shown to be present in the brain, spleen, liver and the lungs (7, 17). It is made up of 165 amino acids of about 18kDa (25), with a number of carbohydrates linked to the polypeptide through O and N glycosidic-bonds giving the glycoprotein a total weight of 34kDa.Two disulphide linkages hold the molecule together (15) and the carbohydrates are responsible for the stability of the glycoprotein in-vivo,and increasing its half-life in the body (24). Why EPO? EPO functions to regulate the amount of red blood cells (RBC) in the blood by controlling the proliferation and differentiation of its immature cells to mature cells (1, 2, 22,). It is also involved in the growth and formation of blood vessels, and healing of wounds (6), it functions in the brain is not clear, but studies showed the glycoprotein to have some protective effects (18). Because of these functions EPO has being used in the treatment of anaemia caused by kidney failure and other causes (25). Recombinant production of EPO Despite its importance, EPO in body is found in very small amounts and mostly in the urine (4), as such there is the requirement to produce EPO in large amounts, this leads to the work of isolating the glycoprotein from the urine (12, 21), and was used to identify its amino acid sequences, and synthesis of its DNA (9, 12), furthermore the human erythropoietin genes were cloned by Lin et al. (17), and consequently recombinant human EPO (rhuEPO) was produced in 1985 using CHO cells (14, 16). Chinese -Hamster- Ovary (CHO-Cells) as rhuEPO host: These are epithelial cells derived from the ovary of Chinese hamster (a mammal). They grow well in culture and looks like cobble stones. The cells usually attach to a surface available but can be grown in suspension (20). CHO cells are grown best at 37oC and at pH 7.4; they are cultured in a suitable complex medium which can support their growth for many generations (20). CHO cell lines are now available from cell culture collections like the American type culture collection; ATCC. Moreover human EPO expression plasmids are now also commercially available, and are usually used for production of EPO using the CHO cells (27). Reasons for choosing CHO-cells Karthik et al. (13) showed that CHO-cells are being used extensively in the industries for the production of many proteins, because they have demonstrated, to possess some qualities like: They can modify biological products post-translationally; Proteins produce in CHO-cells have high glycosylation quality making them compatible and stable (13) Safety of the product; Studies in 1989 have shown that most viruses do not multiply in CHO-cells (13) Ability to adapt easily and be grown in suspension (13). Products can now be purified to contain less contaminant (13). CHO cells have being used for a long time; as such much data has being accumulated for regulatory reasons (13). They are easy to manipulate genetically (13). The isolation of cells deficient in Dihydrofolate-reductase enzymes leads to stable clones selection and genes amplification to increase production (13). With all these, and the fact that EPO is a glycoprotein that requires glycosylation for its stability and activity, recombinant CHO cells are chosen to produce EPO. Cell lines and plasmids: Cell lines which have the capability of glycosylating proteins (Pro-5), harboring the pGEX-HET-puro expression plasmid, will be used to produce the recombinant human erythropoietin (27). Growth strategy Medium: Complex culture medium will be provided with; Glucose as a source of carbon and energy, Amino acids as source of nitrogen, Salts will be included to make the solution isotonic Vitamins and hormones will be added as co-factors Serum is usually added to the culture medium to enhance the growth of the cell (20), but has the following disadvantages: It chemicals are not defined and can cause cell growth inconsistency between batches (20) It is very expensive (20) The serum may contain proteins which can be difficult to separate and purify from the proteins secreted by the cells during downstream processing (20) It increases foaming and can be a source of contamination by viruses. (20) Therefore a serum-free (SF) media (16) will be used for the growth of the E. coli. Process and culture-strategies: The cells will be grown adherent on micro-carriers in a sterile controlled packed bed reactor, and perfusion method of production where some amounts of the medium is removed and replaced by fresh one and the cells are grown slowly will be used (28); because it was found to improve the glycosylation of the proteins more than fed-batch where there is fast growth of cells, (8). Before, many processes were run in a simple batch method, but nowadays, Perfusion or fed-batch methods are mostly employed and higher products are now realized (22). The production will be carried out in two stages; the growth stage and the production stage. Normally stirring will be kept at 100 to 150 rpm, foaming will be avoided by adding Pluronic F68 (16).Temperature will be maintained at 37oC initially during growth and then reduced to 33oC during production, as was shown to increase the overall protein production, while maintaining the quality of the glycoprotein (3, 26). pH w ill be kept at 7.1 initially and then reduced to 6.8 (8, 26), by passing CO2 gas to the culture or by addition of concentrated sodium-bicarbonate solution in low quantities, because CO2 is also toxic to the cells and can also affect the production of EPO (20). In order to avoid the depletion of oxygen, the oxygen transfer rates (OTRs) will be increased above its utilization rate, with a constant supply of pure oxygen and air, while DO will be maintained at 20-50% of air saturation (20). Bioreactor Design: Since the cells are big and fragile, the design of the bioreactor has to be considered. Mammalian cell culture bioreactors are designed with bottoms that are round and are usually made up of glass/stainless steel (20). The impellers are usually marine or pitched blade types fitted at the end of mechanical drives shafts so that both vertical and horizontal mixing are allowed at low stirring-rates (20). Temperature is controlled through coiled pipes or open ended fermenter jacket (20). pH, DO and temperature probes are used for sensing and have both air inlet and outlet for respiration. Growth Analysis Temperature, pH and DO will be monitored on-line, because cells are immobilized, biomass formed cannot be measured directly therefore it will be monitored by measuring rate of glucose consumed daily and the rate of lactate produced (28) Cell viability by flow cytometry, Glucose, glutamine, and lactate concentrations will be analysed using multi-parameter Bio-analytical system (26); while ammonia formed as waste product of amino acid metabolism, will be analysed by colorimetric assay and by the use of detection-kit (26). EPO formed will be analysed using HPLC to determine its purity and its quality by Isoelectric focusing, SDS, and Bradford assay (26). The activity of EPO will be analysed by bioassay and by the use of protein assay-kit (27) Limitations/Problems. There are many limitations associated with CHO cells culture processes and they include; They are fragile and highly sensitive to shear stress caused by agitation and bubble because the cells are large and have only cell membrane (20). This is usually solved using a suitable bioreactor-design and use of Pluronic F68 (20). They need a complex medium including serum which can cause problems in the downstream processing and is expensive (20). Solution to this is by using serum- free media (24, 25). Low yield of proteins have been produced from these cells, the productivity using the microbes being higher than the use of these cells. They also have slow growth rates (13). The problem of low productivity and slow growth rates can be solved through selecting cell lines that are better and optimizing cultural-strategies. Ammonia and lactate are generated during growth and can inhibit growth and also affect glycosylation (8). Solution is by optimizing the strategies of feeding and by monitoring (8). Glycosylation differences may arise from the EPO produced in the CHO-cells and the human EPO as seen in the way the two are sialylated terminally, as a result that the CHO-cells are not able to express an enzyme called alpha-2,6, sialyltransferase (27). Solution is by the use of CHO-cells harboring alpha-2, 6, sialyltransferase-cDNA expression-cassettes (27). REFERENCES: 1. Alcamo, I., DNA Technology; the Awesome-Skill. Farming-dale. New York: Academic Press. (2001). 2. Banting Grolier Electronic publishing www.littletree.com.au/dna.htm accessed on 30/12/2010 3. Carbs information, www.carb-information.com/insulin-synthetic.htm accessed on 30/12/ 2010. 4. Charce, R.E., and Frank, B.H., Research, Production and Safety of Biosynthetic Human Insulin. (1993). www.littletree.com.au/dna.htm accessed on 30/12/2010. 5. Ferrer-Miralles N. Domingo-Espà ­n, J. Corchero, J.L. Và ¡zquez, E. and Villaverde, A. Microb. fact. for recombinant pharmaceuticals, Microbial factories , 8:17, 2009. 6. Fox, S. Improved processes and new capacity for pipeline to commercial production. Biopharmaceutical contract manufacturing, Volume 1 (report). High Tech Business Decisions: San Jose, CA. 2005 7. Genschev, I., Dietrich, G., Goebel, W.,The E. coli alpha-hemolysin secretion system and its use in vaccine development. Trends Microbiol. 10: 39-45. 2002 8. Hewitt C.J., Nebe-von Caron G., Axelsson B., McFarlane C.M, Nienow A.W Studies related to the scale-up of high-cell-density E. coli fed-batch fermentations using multi-parameter flow cytometry: effect of a changing microenvironment with respect to glucose and dissolved oxygen concentration. Biotech. Bioeng. 70: 381-390. 2000 9. Hite P.F, Barnes A.M.J.P.E. Exhuberance over Exubera. Clinical Diabetes 24: 110-114. 2006. 10. Jana, S., Deb, J.K. Strategies for efficient production of heterologous proteins in Escherichia coli. Appl. Microbiol. Biotech. 67: 289-29. 2005. 11. Joseph S., and Raphael F., growing E. coli to high- cell density-A historical perspective on method development Biotech. Advances 23: 345-357 2005. 12. Korz D.J, Rinas U., Hellmuth K, Sanders E.A, Deckwer W.D. Simple fed-batch technique for high cell density cultivation of E. coli. J Biotechnology, 39: 56-65. 1995. 13. Kujau, M.J., Hoischen, C., Riesenberg, D., Gumpert, J. Expression and secretion of functional mini-antibodies McPC603scFvDhlx in cell-wall-less L-form strains of Proteus mirabilis and E. coli: a comparison of the synthesis capacities of L-form strains with E. coli producer strain. Appl. Microbiol. Biotech. 49: 51-58. 1998. 14. Lund, P.A. Microbial molecular chaperones. Advanc. Microbiol. Physiol. 44: 93-140. 2001 15. Makrides S.C. Strategies for achieving high-level expression of genes in Escherichia coli. Microbiol. Rev. 60: 512-5388. 1996. 16. Meyer, H.P. Brass, J. Jungo, C. Klein, J. Wenger, J. and Mommer, R. an emerging Star for Therapeutic and Catalytic Protein Production. Bioprocess International. 2008. 17. Nacelle, G. J. V. and Coppel, R. L. Reshaping Life; Key Issues in Genetic Engineering, Novo-Nordisk Promotional Brochure. Melbourne: Melbourne University Press. 1989. 18. Schmidt, F.R. Recombinant expression systems in pharmaceutical industry. Appl. Microbiol. Biotech. 65:363-37. 2004. 19. Wacker M., Linton D., Hitchen P.G., Nita-Lazar M., Haslam, S.M., North, S.J., Panico M., Morris H.R., Dell A., Wren, B.W., Aeb, M. N-linked glycosylation in Campylobacter jejuni and its functional transfer into E. coli. Science 298:1790-1793. 2002. 20. Demain, L. A., and Vaishnav, P. Production of recombinant proteins by microbes and higher organisms. Biotech.Advan. 27: 297-306. 2009. 21. Schmidt, M., Raman Babu, K., Khanna, N., Marten, S., Rinas, U., Temperature- induced production of recombinant human insulin in high cell density culture of recombinant Escherichia Coli. Journal of Biotech. 68:71-83. 1999. 22. Ratledge, C. and Kristiansen, B. Basic biotechnology. Cambridge: Cambridge university press. 2001. 23. Tabandeh, F., Shojaosadati, S.A., Zomorodipour, A., Khodabandeh, M., Sanati, M.H., Yakhchali, B. Heat induced production of human growth hormone by high cell density cultivation of recombinant E. coli. Biotech. Letters. 26: 245-250. 2004.

Our Town Analysis :: essays research papers

Our Town Analysis   Ã‚  Ã‚  Ã‚  Ã‚  In fact my town does not resemble the setting displayed in this play although I am sure the setting of our ancestors’ towns were much like this one. Back then everybody knew each other and they did not have to lock their doors and they had a strong fear of death and judgement which was indirectly implanted in their religion.   Ã‚  Ã‚  Ã‚  Ã‚  This play was interesting because the representation of death portrayed as being so sudden. When death hit Emily, she suddenly realized how she had wasted her life on earth because they were repeating the same actions everyday of their. She became sad and also regretful that she could not tell her husband George to get on with his life and end this incessant mourning.   Ã‚  Ã‚  Ã‚  Ã‚  The setting is very important because the play is set in a small simple town and it demonstrates the precise actions of people of that time. The setting is special because it is vague therefore it lets your mind imagine the precise details of the scene. This is interesting because you see the scene the way you want it so the author does not need to go into dull descriptions which are not too pleasing.   Ã‚  Ã‚  Ã‚  Ã‚  The atmosphere is very obvious in this play because it is one of the most prominent factors in the play itself. In the first act the dominant mood is happiness because everybody is pursuing their normal activities and gossiping and the first act is used to introduce the characters for us to know their attitudes. In the second act the action is sped up and there is a special event which is marriage involves George and Emily. Emily shows a great deal of hesitation and the attendants at the ceremony are discussing the couple and how they were when they were young. In the third act the mood is sorrow and sadness caused by the death of Emily and her reaction at the overview of her own life.   Ã‚  Ã‚  Ã‚  Ã‚  The relationship between George and Emily is very evident throughout the whole play. In the first act they shared a boy/girl relationship and were good neighbors.

Blind Faith Exposed in The Victim of Aulis :: Victim of Aulis Essays

Blind Faith Exposed in The Victim of Aulis      Ã‚   During World War II, an entire race of people was decimated as a result of blind adherence to one charismatic ruler; the holocaust has become emblematic of the senseless horror of war and the loss of innocent lives. Perhaps influenced by World War II, the Korean War, and the questioning of complete adherence to authority, whose seeds were just breaking through the glorious faà §ade of the 1950's suburban idyll, Dannie Abse wrote "The Victim of Aulis" in 1951-6. The poem is an accusation against the disastrous effects of blind obedience, particularly as it is manifested in religion and war. Abse anchors his critique within the safely distant realm of Greek mythology; this creates a world with which most readers are familiar and thus transfers his indictment of modern society into the images of the cultural psyche. The poet borrows a scene from Greek mythology depicting the sacrifice of Agamemnon's daughter Iphigenia to Artemis at the beginning of the Trojan War, which serves as the ultimate expression of the intimate intermingling of war and religion.      Ã‚  Ã‚  Ã‚   The Greek gods were not only intimately involved in the action of the Trojan War, they were also the impetus for the war. Although the overt cause of the war was Paris' abduction of Helen, this act was the result of quarrelling goddesses. The Trojan prince Paris was forced to choose the fairest amongst the goddesses Hera, Aphrodite, and Athena. Each goddess attempted to sway Paris with offerings, and Aphrodite's temptation was Helen; this leads to the war and the immortal alliances that overshadow its mortal activities. The story that the poem implicitly addresses is of the Achaen king Agamemnon and his daughter Iphigenia. The Achaen forces have gathered at Aulis before mounting their attack on Troy when one of Artemis' stags is killed; this, coupled with Agamemnon's boasting of the act, is why "Artemis is offended" (51). In retaliation, the goddess imprisons the troops at Aulis by preventing the wind from powering their fleet. In order to appease the god dess and begin the war, Agamemnon sacrifices his own daughter Iphigenia as "the child" who will become "the victim of Aulis." Although Artemis intervenes and makes Iphigenia one of her priestesses, only the goddess knows that Iphigenia escaped death.

Cross cultural values and conflicts Essay

The modern society is made up of different cultures which are constantly interacting with each other. This interaction helps in the enrichment of the society. However, it is also the cause of intercultural tensions that have been witnessed in many countries such as the United States. Intercultural conflicts may take different forms. This might lead to problems in different communities and fuel high levels of hatred and confrontation. It is a known fact that cultures differ from one community to another. There has been increased violence that can be attributed to marginalization and impoverishment of some cultures as compared to others. The other factors that contribute to this are ignorance coupled with prejudice. The result of this is disagreements between different communities, resentment and possibility of uncontrollable violence erupting. These conflicts arise due to opposition of certain cultures and reluctance to accept the diverse cultures of the world. Sources of conflicts Intercultural conflicts can, therefore, be said to be due to three causes. These are: political causes, social causes and economic causes. Political sources may be due to territorial differences that might lead to conflicts between different groups of people or nations. The fight for certain resources in particular regions fall under economic causes of these conflicts. Dispute may sometimes arise regarding the ownership, accessibility to or control of certain resources. These resources might include jobs, contracts, credits or education. Allocation of these resources should be done in a fair manner so that everyone gets an equal share regardless of their cultural backgrounds. The tough economic times and conditions may exacerbate intercultural hostility especially when these are seen to be the key causes of unemployment and degradation of peoples’ welfare (LeBaron and Pillay, p 42). Economic policies that favor certain groups of people or nations and ignoring the disadvantaged ones may hasten these conflicts. Such disadvantaged groups include immigrants and workers who might be looked down upon (LeBaron and Pillay, p 42). Social causes of conflicts might be due to differences in religion and languages. These are cultural issues which should not be taken lightly. For example, a communication barrier as far as language is concerned. This can be seen in institutions regarding the particular language used in the teaching process and examinations. Secondly, language used in the military during command and other government departments for communication. These might cause disparities between people of different ethnic and cultural backgrounds. It has been established that religion is a major cause of conflict between groups of people (LeBaron and Pillay, p 43). The main cause of social conflicts in the U. S. is immigration and the income levels between the two races (whites and blacks). This also includes the natives and immigrants in the United States as well as the poor and rich people (Morin, para 2). Other factors that may worsen this situation include; potential threats posed by certain groups to the interest of the group considered as the majority. For example, in the United States, most whites see the non white immigrants as criminals who pose a great danger to their families, jobs and institutions. This leads to exclusion of such migrants in certain sectors such as taking up high grade jobs and government positions. It has heightened the levels of discrimination in the United States (Ting-Toomey and Oetzel, p 23). Conclusion There is need for a global approach in finding a lasting solution to this problem. These efforts should be directed towards ending cultural, racial and ethnic conflicts worldwide. It will help in ending xenophobia, racism and racial segregation in the U. S. In doing so, the tension that is usually witnessed between different groups of people will be greatly reduced hence leading to a harmonious society. Despite the fact that conflicts will always arise at times, these should be solved amicably and democratically without bias or favoring any side or group. Works Cited LeBaron, Michelle and Venashri Pillay. Conflict across Cultures. Boston: Intercultural Press, 2006. Morin, Rich. â€Å"What Divides America? † September 24, 2009. August 10th, 2010 Ting-Toomey, Stella and John Oetzel G. Managing Intercultural Conflict Effectively. California: Sage Publications, 2001

Akdj

Learning Module 3 Connor MacLeod 100254620 The first video, The Solar Nebular Theory, describes the planetary formation through a collapsing cloud of dust and gas that spins in the same direction at a fast pace. Through angular momentum, as it collapses even more, it speeds up and flattens into a disk. As it flattens, the materials start to come together and planets start to form as well as a young star is created in the middle. What I liked about the video is how they used good analogies such as the pizza and the figure skater to get the idea of angular momentum across.What I didn’t like about the video is how they didn’t go more in depth about the other bodies that are created around the star. To make it better I would’ve used our solar system as an example in order to relate to it better The 2nd video is the Planetisimal video. It goes on to describe how particles grow by condensation, the process of adding matter to an atom or molecule. Once they get to a big enough size, the particles start to stick to others through a process called accretion. Planetisimals continue to grow by colliding and sticking to others until it gets large enough to turn into a protoplanet.The stage where iron falls to the core and heat is generated while lighter materials stay on the outside to create the earths crust. What I did not like about the video is how they do not specify how long it takes for it to turn into a protoplanet from a planetisimal. The woman talks about millions of years to get to the diameter of the particle to centimeters until they start to collide, but lacks the information of how long the collisions take to get bigger. They could improve the video by adding these details into it.What I liked about the video is how it goes in depth about protoplanets and the gravitational collapse as the textbook had me somewhat confused. The 3rd video is the extra solar planets. It is about how to detect planets outside of our solar system that gravita te around other stars. They don’t try and find planets directly, they observe the stars in which they believe planets gravitate around and watch how the star reacts. The way they can measure this is through the Doppler affect. As the planet gravitates around it, it pulls the star in different directions.So when a star comes towards you, its light gets compressed, and when it is moving away, the light waves get stretched. What I liked about the video is the way they illustrated how the Doppler affect worked with visuals as oppose to the dog theory in the textbook. That being said, what I didn’t like about the video was how they focused solely on detecting planets through the Doppler affect, I would improve upon this video by not leaving out the other approach of detecting the difference in light when a planet moves in front of it.The 4th video is about meteorites. It discuses how meteoroid’s cause meteors, that is until it reaches the ground and are renamed to me teorites. It then describes how each meteorite is classified by its composition. Each different composition inside a meteorite allows geologists to identify their space origins. What I liked about the video is how they discussed the different kinds of meteorites and the significance of different compositions of the meteorites. What I didn’t like is how they did not touch upon meteor showers and different meteor showers.To improve this video I would’ve included some facts of the Perseid meteor shower. The 5th video is about asteroids. It discusses how Italian astronomer Piazzi discovered the first asteroid. Now there are over 100,000 asteroids that lay in the asteroid belt near mars and Jupiter. They also describe the reason the asteroids are in the belt could be due to a planet that never formed. The pull of Jupiter’s gravity kept the particles accelerating so fast, they could never accrete.What I liked about the video is how it spent the most time discussing wh y the asteroid belt exists as that caught my attention the most in the textbook, where its only mentioned briefly. What I didn’t like about the video is it didn’t discuss the shapes and size of the asteroids themselves. To improve this video I would’ve added some interesting facts from the textbook such as some are asteroids are 200km in diameter. The 6th video is about comets. It talked about how civilization was influenced from comets and the association of comets to earth through eligious or mythological oriented societies and how they were viewed as a bad omen. What I did not like about this video is how it did not describe at all what a comet is, or what it is made up of. They could make this video better by discussing the basics of a comet, not just the historic significance it played on our societies. That being said, what I did like about this video is the telling of how it played a role in our societies that if something bad happened, and a comet was in the sky, it started to be associated with each other.

Negotiation Between Countries

I want to do that from three different fronts, namely; Cultural differences in communication The patterns of communication between the countries are different even though tooth countries communicates basically in English language as an accepted language of communication. Communication in this sense can be verbal or non-verbal. Non- verbal also Includes written communications. Nigeria has over 400 languages but English language Is the accepted lingua franca. The united States has other native languages but English Is still the accepted language of communication.Though I will communicate in English with my American business partners but I must also understand that the way the Americans write and pronounce some words is different from the way we write and pronounce our words in English language. The American counterparts for instance will write the word â€Å"favor as favor† but I will write the word â€Å"favor as favor† because Nigeria was colonized by the British, so we take our language after the British pattern of writing and communicating.Nigerian are exclusion minded in communication; we will tend to see how the conversation does not align with what we already know and would usually want the American counterpart to align with our own position of knowledge. The American partner will rather communicate based on an Inclusive mindedness; they will rather want to know how what I am saying makes sense. American look for the sense In the conversation, does the numbers tally, are the figures correct, so they have a broader view to the conversation.Also Most Nigerian communication style follow after the collectivists' culture; (Grove & Hallowed; 1994) â€Å"Collectivists convey negative feedback indirectly via an intermediary or by quiet withdrawal of a perquisite. Sometimes they omit saying anything that's negative. If you are managing employees in (or from) a collectivist culture, you might not find out what's going wrong in your operation until the proverbial eleventh hour; there's deep reluctance to upset group harmony by ailing you the bad news†.This means we would rather not say â€Å"no† to you openly even though we disagree, but we will rather let the conversation go on peaceably until we have another time to object to most of the Initial points of our conversation. My American partner would rather tell It the way It Is If It doesn't go down with them. (Grove & Hallowed; 1994) They pattern after the Individualist culture In conversations. Generally speaking, American culture communication style are usually Comfort talks, they are emotionally objective and practical in approach, their interactions are usually very direct and open.They don't hide any cards off the table, they play all the cards on the table. But when it comes to reactions, they react to issues analytically and they remain persistent in their analysis until otherwise convinced. These are the factors that influences how Americans communicate. Cu ltural differences in negotiation and conflict-resolution Let's think of a scenario of negotiation with our U. S. Partner for a presentation from an IT company overseas. The American makes his presentation very apt and technically done by expressing his overwhelming objective points and reasons why the product is best for my company in Nigeria. Well, we would say â€Å"the presentation is brilliant but I still do not trust this guy. He thinks objectively while I think subjectively, so our negotiations can either stall or proceed based on how we handle this cultural differences between our cultures. (Mimicked, 2010) To most Americans, difference is a threat, they will naturally tend to overlook similarities and take note of the difference when they first begin the negotiations. We Nigerian are looking for similarity with what we already know and experience, similarity makes us comfortable in the negotiation.During negotiations with Americans, you must understand that they hate silen ce, and they would often interrupt you at intervals during negotiations, while that is rude in my country, to interrupt someone while he is still talking, the Americans have that as a normal way of life. Cultural differences in problem-solving and decision-making In terms of problem solving, the American approaches it from a highly objective point of view, they would naturally first carry out a critical analysis of the problem and then cake decisions based on the facts on ground.While we Nigerian usually make decisions based on sentiments sometimes, we decide to enter a business agreement when we feel intuitively inclined to do so, there is mostly no objective reason except we feel convinced inside that this is a good buy so we make the money decision. I would first seek an understanding of the cultural differences that have been highlighted earlier. After which I will approach the negotiation objectively, knowing that is how my American partner thinks.I will also approach most of o ur negotiation room an individualist point of view while also letting my American friend understand the collectivist point of view. My approach would be to achieve a win-win situation. In my country I can afford to have a win-lose situation during negotiations but I understand that the American is highly objective and would not go by my sweet talks or by my expressing an overwhelming emotional conviction about the business I am trying to sell to him. So my thought pattern will naturally change from when negotiating with my fellow country man.The factors of negotiation and business allegations to consider in this case would be; Listening skills- I will need to be more active in listening so that I get the objective point the American is making rather than rely on my emotions to guide my decision making in the negotiation. People skills – I need to feel comfortable with our differences and not look hard for similarities between us before I connect with the American counterpart. BATAAN – I must establish what the Best Alternative To Negotiated Agreement is. This is vital to my than what I may have done otherwise. The BATAAN is what I can or might do if an agreement cannot be reached.This is my final position in case an agreement is difficult to reach. Have a Plan – I would consider having a plan ahead of the negotiation. My plan would likely include the following: (Yachted, D; ND) Trying to establish the negotiating style of the other party. This helps me think through how best to communicate and go through the process of negotiation and making educated guesses as I go along. What are my interests? This is gives me a perfect reason to answer the question of why? Before the negotiation. What is my real interest? What are the interests of the American Partner?I must understand where y American partner is coming from what are the shared interests we have and what are the opposing interests. Opposing interests is what I must negotiate. What do I have that I can trade that is of lesser value to me and of higher value to the American Partner? This helps me consider the options in the give and take phase of the negotiation. I must know beforehand what I can trade off at any time during the negotiation that is of a lesser value to me and of a higher value to my partner. What are three options I can implement to move the negotiation from compromising to joint problem solving?

Bourgeoisie & money

The pictures represent the emotion of the people. Not only are they struggling to maintain what they have and who they are – they are still seeking. In their eyes they are looking for a way – a way forward – or a way out of their situation. The images reflect that control the Bourgeoisie had over the working class. They controlled the money and the industry. Those who did the labor to make the production levels – received very little reward for their efforts. Neither monetarily or in satisfaction of what they were doing. This is reflected in many of these pictures.A person in the industrial working class often lived a life of want. Never having enough income or wealth to move out of the social class they were in. Resentment was a common emotion towards the Bourgeoisie from the industrial working class. This emotion is what Marxism used to build a movement around, The working class felt there was nothing they could do to get up to the next level of what the y perceived as social classification. The latter nineteenth century gave birth to a generation who was resentful of where they were at in social class.A generation who wanted more than the Bourgeoisie were willing to part with. The Bourgeoisie had control – and were very protective of the control they enjoyed. The working class was hoping for a way out of their situation – they wanted more control and more freedom that increased control and wealth could give them. The pictures identify clearly the emotion of this segment of society – and the yearning for something different. There are no answers in their eyes, only questions and confusion. The answers and clarity they were seeking was still a generation away.