Oxidative Stress and Diabetic Nephropathy The WritePass Journal

Oxidative Stress and Diabetic Nephropathy Introduction Oxidative Stress and Diabetic Nephropathy IntroductionEconomic burden of Diabetes for families and society:Causes of microvascular damage in diabetes:Advanced glycosylated end-products (AGEs):Oxidative stress and Reactive Oxygen Species (ROS):Polyol pathway/aldose reductase:Hexosamine pathway:Endothelial dysfunction pathogenesis:REFERENCES: Related Introduction Diabetes mellitus is a chronic non-communicable condition resulting in high levels of glucose in the blood.   It occurs due to inability of the beta cells in pancreas islet tissue to produce enough insulin, or when the body becomes resistant to insulin.   It reduces both quality and length of life and over time leads to serious complications such as coronary heart disease, stroke, neuropathy, retinopathy and nephropathy. There are two main types of diabetes mellitus: Type 1 diabetes: also called insulin-dependent diabetes mellitus (IDDM), early-onset and juvenile diabetes. It is an autoimmune disease and results from destruction of insulin producing beta cells in islet tissues of pancreas by the body’s immune system. The subsequent lack of insulin results in high blood glucose levels, which if not controlled by exogenous insulin results in multiple organ damage. Type 2 diabetes:   formerly called non-insulin-dependent (NIDDM) and adult-onset.   It is a metabolic disorder that mainly occurs in individuals over the age of 40.   In this type of diabetes high blood glucose results either due to relative insulin deficiency or insulin resistance.   Lifestyle and genetic factors play an important role in the development of type 2 diabetes. Type 2 diabetes is a growing problem among the elderly population and is widely predicted to grow in the future.   Since the population is aging in the western world, so it is not surprising that elderly population will contribute to future increase but other factors such as lifestyle and diet will also play a major role. WHO survey 2010 estimated that 285 million of the world’s population have diabetes and more than 70% of them live in low and middle income countries. It is also estimated that this burden will increase to 438 million by 2030 (Diabetes fact, 2011). Wild 2004 projected that the total number of individuals with diabetes worldwide will increase from 171 million in 2000 to 366 million 2030.   Although the prevalence of diabetes is higher in men compared to women but there are more women with diabetes than men.   In developing countries type 2 diabetes mainly affects people of working age, between 35 and 64 years, whereas in developed countries the majority of people with diabetes are above the age of retirement i.e. above 65 years of age (WDD06 – Karachi, 2006).   India has the largest diabetes world’s population i.e. 50.8 million followed by China with 43.2 million ( Express news report,   2009). In Europe prevalence of clinically diagnosed diabetes was estimated to be 3% in 1997.   It was estimated to increase to around 3.6% by 2000 and to over 4% by 2010 (Scottish Diabetes Survey 2003).   In UK 4.26 % of population has diabetes according to the Diabetes UK statistics (Diabetes prevalence 2010).   Scottish Diabetic Survey, 2010, projects that 4.6% of Scotland population has diabetes out of which 87.7% have type 2 diabetes. Diabetes is also at an increase among children. Diabetes amongst children is primarily Type 1 diabetes but Type 2 diabetes is also increasingly being diagnosed. One of the major contributing factors for this rise in diabetes among children is the increase in the number of children who are overweight or obese. â€Å"Twenty five children in every 100,000 in Scotland have diabetes, compared to 17 in England and Wales.†Ã‚   An increase in this at a rate of 2% per years has been suggested by Diabetes UK in Scotland, as a result tripling of new cases in the last 30 years has been seen (ABPI Report Scotland, 2005). Economic burden of Diabetes for families and society: Diabetes and its complications have a significant economic impact on individuals, families, health systems and countries. For example, WHO estimates that in the period 2006-2015, China will need to allocate $558 billion in foregone national income due to heart disease, stroke and diabetes alone and India will spend $336.6 billion (Diabetes, 2011). â€Å"In the poorest countries, people with diabetes and their families bear almost the whole cost of the medical care they can afford.†Ã‚   In Latin America, 40-60% of medical care expenditures is paid by the families themselves. In Mozambique, 75% of the per capita income is spent on diabetic care by one person; in Mali it amounts to 61%; Vietnam is 51% and Zambia 21%. It is estimated that poor people with diabetes in some developing countries spend as much as 25% of their annual income on private care (Diabetes fact. April 2011). The trend of diabetes in developing countries show that it mostly affects working age group, between 35 and 64 years, relative to developed countries where the majority of diabetes population are aging. Therefore when principal wage earner is affected by diabetes and its complications, the choice between healthcare expenses and food or clothing can trap the whole family in a downward spiral of worsening poverty and health. According to WHO, an annual 2% reduction in chronic disease death rates in Pakistan would provide an economic gain of 1 billion dollars over the next 10 years (WDD06 – Karachi, 2006). The cost incurred by diabetes morbidity are far greater than the cost of the disease prevention (Editorial in Lancet:   World Diabetes Day 14th November, 2010). In a press report by the independent economic consultancy group NERA it is assessed that intensive management of Type 2 diabetes in Scotland can decrease hospital cost by  £41 million by saving over 91,000 bed days a year in 2025 and will also save  £78 million a year in lost work days (ABPI Report Scotland, 2005). Diabetic Nephropathy: The diabetic complication, nephropathy is a condition with high unmet therapeutic needs.   It is linked with significant increases in morbidity and mortality risk, and is the most common cause of ESRD in the Western countries. Diabetes-induced damage in the kidney leads to microalbuminuria. This progresses to ESRD, which requires dialysis or transplantation. Diabetes accounts for over 40% of ESRD (Diabetic Nephropathy, 2003). The main focus of therapy in diabetic nephropathy is on tight control of blood pressure. Guidelines have progressively revised the target BP goal downwards, currently at 125/75 mmHg in patients with 1g proteinuria, and now recommend either ACE or ARB (Diabetic Nephropathy, 2003). In the U.S., diabetic nephropathy accounts for about 40% of new cases of ESRD.   In 1997 the cost required for treatment of diabetic patients having ESRD amounts to $15.6 billion. There is considerable racial/ethnic variability in this regard, Native Americans, Hispanics (especially Mexican-Americans), and African-Americans have much higher risks of developing ESRD than non-Hispanic whites with type 2 diabetes (Mark, 2001). In the UK, 1,000 people with diabetes start kidney dialysis every year. (Diabetes in the UK, 2004). Ahmedani 2005 reports that in Karachi, Pakistan overall prevalence of microalbuminuria was found to be 34% in patients with diabetes and this was strongly associated with the age, diastolic hypertension, diabetic retinopathy and serum low density lipoprotein. End stage renal disease is a most serious complication of diabetes and accounts to be the most expensive for NHS. Diabetic nephropathy usually develops 15-25 years after the occurrence of diabetes.   In Scotland, 20% of patients who undergo renal transplantation are diabetic. In diabetic individuals, microalbuminuria and stroke, or an increased serum creatinine levels raises the risk of renal nephropathy and failure.   Poor glycemic control and high blood pressure are risk factors of diabetic renal disease (Scottish  Diabetes  Framework, 2002). Diabetic renal impairment is a strong indicator of Cardiovascular disease and cardiovascular disease is the major cause of morbidity and mortality is diabetic patients (Guillausseau, 2011).   Annual cardiovascular mortality is 0.7% in normoalbuminuric patient as compared to 2% in microalbuminuric patients and 12% in the patient with elevated creatinine (Stratton IM, 2009) In a review by Vishwanathan, 1999, it is explained that South Asians and Afro-Caribbean are more susceptible to develop renal disease relative to European. Retinopathy increases the risk of diabetic nephropathy. Prevalence of diabetic nephropathy in India was 30.3% in a study done among 4837 patients with chronic renal failure over a period of 10 years. He further argued that an increased prevalence of microalbuminurea among South Asians having type 2 diabetes mellitus relative to Europeans by 1.2 (men) and 1.7 (women) folds.   According to SIGN 116, the incidence of diabetic nephropathy in patient with type 1 diabetes can be considerably reduced by attaining good glycaemic and tight blood pressure control.  ­ In a report by Singh NP, 2003, it is suggested that the incidence of diabetic kidney disease can be reduced by:   tight blood glucose control, blood pressure control, rennin-angiotensin-aldosterone system blockade and protein restriction. Causes of microvascular damage in diabetes: Long standing hyperglycemia lead to a number of damages including: Advanced glycosylated end products (AGES) Oxidative stress Increased sorbitol (polyol pathway) Increase in hexosamine pathway Impaired endothelial function Immune effect All these damages result in microvascular complications of diabetes. Advanced glycosylated end-products (AGEs): Chronic hyperglycemia causes increased glycosylation of proteins leading to AGEs, which in turn results in loss of structure and function, turning on/off signal pathways within cells and alteration in gene expression. AGEs are sugar-derived compounds, glucose binds amino groups on proteins, lipids and nucleic acids to form AGEs.   AGEs form at a constant but slow rate throughout your life (even as an embryo) (Peppa et al, 2003). AGEs interact with RAGE (surface AGE-binding receptors) resulting in proinflammatory effects, formation reactive oxygen species, loss of oxidants (oxidative stress) and altered gene transcription. Levels of AGEs relates to extent of microvascular complications in diabetes.   AGEs contributes to atheromatous plaque by stimulating low-density lipoprotein (LDL) oxidation and the deposition of oxidized LDL. AGEs leads to endothelial dysfunction, macrophage activation, and impaired vascular smooth muscle cell function.   Experimentally, AGEs cause glomerular damage and proteinuria. Oxidative stress and Reactive Oxygen Species (ROS): Oxidative stress is an imbalance between ROS production and antioxidants.   Oxygen is used by cells to carry out their normal functions and as a side effect produces free radicals.   Free radicals are missing an electron so are unstable and highly reactive.   Free radicals steal electrons from molecules within cells causing oxidative damage to proteins, membranes and genes. Polyol pathway/aldose reductase: Aldose reductase (AR) normally reduces toxic aldehydes into inactive alcohols inside the cells. Glucose perfuses into some cells without insulin e.g. nerves.   During hyperglycaemic condition, AR reduces that excess glucose to sorbitol (a polyol).   Polyols are trapped inside the cells creating an osmotic gradient.   Sodium and water flow into the cell resulting in oedema. But sorbitol can be metabolised to fructose by the actions of sorbitol dehydrogenase.   High fructose leads to AGEs resulting in more cell damage (Takaqi et al, 1995). Hexosamine pathway: Glucose is mainly metabolised through glycolysis, some gets diverted into an alternative pathway, ending up as UDP (urine diphosphate) N-acetyl glucosamine.   This alters transcription factors, often leading to pathologic changes in gene expression e.g. increased expression of transforming growth factor-B1 and plaminogen activator inhibitor-1, which damages blood vessels. Endothelial dysfunction pathogenesis: Hyperglycemia leading to the formation of AGEs, ROS, the glycosylation of proteins and increased inflammatory cytokines etc.   As a result   small blood vessels, particularly the endothelium are damaged causing vasoconstriction, ischemia, and reduced flow to tissues that rely on the vessel for oxygen and nutrients. Growth factors are also released leading to the blood vessel wall thickening and occlusion of small blood vessels. Nerve growth factors (NGF) and factors like it are damaged.   These factors keep nerves healthy and capable of re-growth if damaged. Changes to the immune system lead to release of toxic cytokines, blockage of blood vessels with leukocytes and loss of normal immune cell action. In this dissertation, a recent aspect of one of the above causes of microvascular damage of diabetes leading to nephropathy will be considered. Current studies have uncovered new insights in the role of oxidative stress in diabetic renal disease, suggesting a different and innovative approach to a possible â€Å"casual† antioxidant therapy. In this dissertation the role oxidative stress may play in the development of diabetic kidney disease will be discussed.   The role of antioxidant therapy in managing or delaying the progression of diabetic nephropathy will be addressed. REFERENCES: Ahmedani M Y, (2005) Prevalence of Microalbuminuria in Type 2 Diabetic Patients in Karachi: Pakistan A Multi-center Study: jpma.org.pk/full_article_text.php?article_id=856 ABPI Report Scotland, (2005)The future burden of CHD and Diabetes in Scotland: The value of health care innovation. Available at: s3.amazonaws.com/zanran_storage/www.abpi.org.uk//50031328.pdf Diabetes fact. (2011) Available at: worlddiabetesfoundation.org/composite-35.htm Diabetes (2011), Available at: who.int/mediacentre/factsheets/fs312/en/ Diabetic Nephropathy 2003. Available at: datamonitor.com/Products/Free/Brief/BFHC0625/010BFHC0625.pdf Express news report, India has largest number of diabetes patients: Report (2009) indianexpress.com/news/india-has-largest-number-of-diabetes-patient/531240/ Diabetes in the UK 2004, www.diabetes.org.uk/Documents/Reports/in_the_UK_2004.doc Guillausseau, (2011)   Type 2 diabetes and cardiovascular risk: kidney function is pivota. Available at: diafocus.com/2011/01/11/type-2-diabetes-and-cardiovascular-risk-kidney-function-is-pivotal/ Peppa M, Uribarri J, Vlassara H, 2003, Glucose, Advanced Glycation End Products, and Diabetes Complications:   What is New and What Works. Available at: http://clinical.diabetesjournals.org/content/21/4/186.full Stratton IM, (2009) Association of glycaemia with macrovascular and microvascular complications of type 2 diabetes (UKPDS 35): Prospective observational study   Available at: http://articulos.sld.cu/medicinainterna/files/2009/10/association-of-glycaemia-with-macrovascular-and-microvascular.pdf Scottish  Diabetes  Framework. (2002) Available at: scotland.gov.uk/Publications/2002/04/14452/1986 Scottish Diabetes Survey 2003, Available at: scotland.gov.uk/Publications/2004/10/20023/44203 Singh NP, Singh D, 2003, Diabetes Mellitus – An Overview For Family Physicians. Available at: http://delhimedicalcouncil.nic.in/diabetes-mellitus.html Sign 116, Available at:   sign.ac.uk/pdf/sign116.pdf Scottish Diabetes Survey 2010, Available at:   diabetesinscotland.org.uk/Publications/Scottish%20Diabetes%20Survey%202010.pdf Takaqi Y, Kashiwaqi A, Tanaka Y, Asahina T, Kikkawa R, Shigeta Y, 1995, Significance of fructose-induced protein oxidation and formation of advanced glycation end product.   Available at: ncbi.nlm.nih.gov/pubmed/7599353 Viswanathan V, (1999) Type 2 diabetes and diabetic nephropathy in India- magnitude of the problem.   Available at: http://ndt.oxfordjournals.org/content/14/12/2805.full WILD S et al, (2004) Global Prevalence of Diabetes.   Available at: who.int/diabetes/facts/en/diabcare0504.pdf WDD06 – Karachi, (2006). Diabetes kills without distinction. Available at: idf.org/wdd06-karachi

Civil Wars Wet Plate Collodion Photography

Civil War's Wet Plate Collodion Photography The wet plate collodion process was a manner of taking photographs which used panes of glass, coated with a chemical solution, as the negative. It was the method of photography in use at the time of the Civil War, and it was a fairly complicated procedure. The wet plate method was invented by Frederick Scott Archer, an amateur photographer in Britain, in 1851. Frustrated by the difficult photography technology of the time, a method known as calotype, Scott Archer sought to develop a simplified process for preparing a photographic negative. His discovery was the wet plate method, which was generally known as the â€Å"collodion process.† The word collodion refers to the syrupy chemical mixture which was used to coat the glass plate. Numerous Steps Were Required The wet plate process required considerable skill. The required steps: A glass sheet was coated with chemicals, known as collodion.The coated plate was immersed in a bath of silver nitrate, which made it sensitive to light.The wet glass, which would be the negative used in the camera, was then placed in a light-proof box.The negative, in its special light-proof holder, would be placed inside the camera.A panel in the light-proof holder, known as the dark slide, along with the lens cap of the camera, would be removed for several seconds, thereby taking the photograph.The â€Å"dark slide† of the light-proof box was replaced, sealing the negative up in darkness again.The glass negative was then taken to the darkroom and developed in chemicals and â€Å"fixed,† making the negative image on it permanent. (For a photographer working in the field during the Civil War, the darkroom would be an improvised space in a horse-drawn wagon.)The negative could be coated with a varnish to ensure the permanence of the image.Prints would later be generated from the glass negative. The Wet Plate Collodion Process Had Serious Drawbacks The steps involved in the wet plate process, and the considerable skill required, imposed obvious limitations. Photographs taken with the wet plate process, from the 1850s through the late 1800s, were almost always taken by professional photographers in a studio setting. Even photographs taken in the field during the Civil War, or later during expeditions to the West, required the photographer to travel with a wagon full of equipment. Perhaps the first war photographer was a British artist, Roger Fenton, who managed to transport cumbersome photographic equipment to the battlefront of the Crimean War. Fenton had mastered the wet plate method of photography soon after it became available and put it into practice shooting landscapes of the British midlands. Fenton took a trip to Russia in 1852 and took photographs. His travels proved that the latest photographic method could be utilized outside of a studio. However, traveling with the equipment and the necessary chemicals to develop the images would present a formidable challenge. Traveling to the Crimean War with his photographic wagon was difficult, yet Fenton managed to shoot impressive photographs. His images, while praised by art critics upon his return to England, were a commercial failure. Roger Fentons photographic van used in the Crimean War, with his assistant posing on its bench. Library of Congress While Fenton had transported his ungainly equipment to the front, he purposely avoided photographing the ravages of war. He would have had many opportunities  to depict wounded or dead soldiers. But he probably assumed his intended  audience in Britain did not want to see such things. He sought to portray a more glorious side of the conflict, and tended to photograph officers in their dress uniforms. In fairness to Fenton, the wet plate process made it impossible to photograph action on the battlefield. The process allowed for a shorter exposure time than previous photographic methods, yet it still required the shutter to be open for several seconds. For that reason there could not be any action photography with wet plate photography, as any action would blur. There are no combat photographs from the Civil War, as people in the photographs had to hold a pose for the length of the exposure. And for photographers working in battlefield or camp conditions, there were great obstacles. It was difficult to travel with the chemicals required for preparing and developing the negatives. And the glass panes used as negatives were fragile and carrying them in horse-drawn wagons presented a whole set of difficulties. Generally speaking, a photographer working in the field, such as Alexander Gardner when he shot the carnage at Antietam, would have an assistant along who mixed the chemicals. While the assistant was in the wagon preparing the glass plate, the photographer could set up the camera on its heavy tripod and compose the shot. Even with an assistant helping, each photograph taken during the Civil War would have required about ten minutes of preparation and developing. And once a photograph was taken and the negative was fixed, there was always a problem of a negative cracking. A famous photograph of Abraham Lincoln by Alexander Gardner shows damage from a crack in the glass negative, and other photographs of the same period show similar flaws. By the 1880s a dry negative method began to be available to photographers. Those negatives could be purchased ready to be used, and did not require the complicated process of preparing the collodion as required in the wet plate process.

Globalization and the maritime world in the twentieth century Essay

Globalization and the maritime world in the twentieth century - Essay Example Marc indicates that cargo ships were the major systems of transport and were used load and offload goods (166). The goods in this context had different destinations throughout the world. This concept simply explains that globalization was taking place as the goods were sent to different places. The world was soon becoming smaller thanks to the development in the maritime industry. Marc explains that the shipping lines ordered for larger ships that would be used transport the aluminum boxes that were the standard containers (202-21). Additionally, the development of the maritime industry saw the modernization of port cities to accommodate the larger ships (Marc 191-201). The author also argues that in the 1980s computers ventured in as well, that led to a containerized system (Marc 125). A closer analysis of this fact would be said to have shaped the global economy through innovations in the maritime industry. Miller backs the work of Marc by arguing that globalization is historical and can be rooted to the reality of shipping and trading. The maritime world, in his thought, was the major reason behind the global interconnectedness (Miller 11). In the twentieth century, for example, there were a lot of Eurocentric exchanges in the sea. There was the existence of a global maritime system where European ports were constructed, shipping lines as well as partial networks (Miller 12). This commercial maritime world played a huge role in shipping the present day world of business. The Europeans were the principal operators of the global trading and transport systems up to the 1960s. Just like Marc, Miller would argue that there is a great link between containerization, trade, de-colonization and economic growth thanks to the maritime industry and its developments. Through the works, the close correlation between the maritime world in the 20th century and globalization is evident. The process of globalization clearly went through progressions that saw the

Kingdom of Heaven Essay Example | Topics and Well Written Essays - 1000 words

Kingdom of Heaven - Essay Example For example, if a film is set in the time of Christ, the people need to be following the customs of the Roman Empire along with wearing the correct robes. They should not be in a three piece suit and penny loafers, unless the movie has to deal with a man out of time. The third responsibility of the filmmaker is to not keep the locations to their accurate size so that the viewer seems as though they were there. The historical piece that will be used to determine the three responsibilities is Ridley Scott’s 12th Century epic, Kingdom of Heaven. Kingdom of Heaven is a film set during the Crusades about a French blacksmith named Balian, who is searching for a reason to go on after the death of his wife and children. A fabled knight, Godfrey of Ibelin, has briefly returned home after serving in thr East. Godfrey approaches Balian and let’s the blacksmith know that he himself is the blacksmith’s true father. Godfrey, then, asks Balian to join him and his troops in thei r journey to return to the Holy City of Jerusalem to help in the city’s defense. The blacksmith accepts the offer of Godfrey. Their arrival falls in between the Second and Third Crusades when Jerusalem is enjoying a period of peace between the Muslims and the Christians. This peace was all thanks to the Christian monarch King Baldwin IV, his second-in-command Tiberius, and the Muslim potentate Saladin. Unfortunately, the peace does last since violent agitators set out to increase their power. Saladin had to bow down to the pressure Godfey and his men stayed o give their allegiance to the king and his community of diversity. The knights, as well as Balian, use their skills as warriors to build a lasting peace. Orlando Bloom (Balian) is one of the main stars of the film along with Liam Neeson (Godfrey), Edward Norton (Baldwin IV), and Jeremy Iron (Tiberias). In Kingdom of Heaven, I noticed that the overall peace between the Muslims and Christians seemed to show the lack of reli giousness. Rather, the situational tension amongst the Christians and the Muslims seem to be more like a backdrop, than a major part of the story. The story is set between the Second and Third Crusades. However, the action and fighting as warriors is more prevalent than the religious aspects of the Crusade that should be more available. King Baldwin IV was a monarch who wanted to convey peace and diversity amongst the religious factions that want to control the Holy City of Jerusalem. Thus, the filmmaker seemed to use the facts of the time period as more of a symbol for the setting, than as the setting itself. The fighting may have been more gruesome for a film that should be more focused upon Baldwin’s Kingdom of Heaven, then the relationships of a lone person. The relationships should be more about the groups trying to vie for power in Jerusalem, than that of a single man. This story portrays single man being effective in the war to maintain the peace and diversity througho ut the city of Jerusalem during the reign of Baldwin IV. However, the portrayal should be how Baldwin could bring about the change in the ancient fight of the Muslims and the Christians, not how a French blacksmith becomes a knight to help defend the ancient, Holy City from the agitators that wish to overthrow Baldwin IV and claim power for themselves. I feel the overall responsibility of keeping the accuracy of the facts was a little massacred instead of being kept intact by the screenwriters or even portrayed effectively by

Plasmid Extraction :: essays research papers

Introduction   Ã‚  Ã‚  Ã‚  Ã‚  Chitobiase, from Vibrio harveyi, is a membrane bound lipoprotein involved in the degradation of chitin. Chitobiase is similar to and may share a common ancestry to the a-chain of human b-hexos-aminidase. Chitobiase is encoded by chb.   Ã‚  Ã‚  Ã‚  Ã‚  In this experiment, a restriction map for restriction enzymes Eco R1, Pst1 and Hind III using Southern hybridization and restriction analysis of pRSG 192. pRSG 192 is a recombinant plasmid derived from the chb gene and pUC 19, a 2.7kb engineered plasmid which encodes for ampicillin resistance, a portion of the lac operon and a multiple cloning region . The chb gene exists as a 3.6 kb insert in the mutiple cloning region of pUC 19.   Ã‚  Ã‚  Ã‚  Ã‚  The major goals of Experiment One will be to isolate pRSG 192 from an overnight culture of E. coli, amplify a region of the chb gene using PCR, and to map restriction sites within the chb gene using restriction analysis and Southern hybridization. Methods Plasmid Isolation Four microfuge tubes containing cell pellets representing 3.0ml of cells(2 x 1.5ml) from an overnight culture of E. coli were prepared. The supernatant fluid was discarded and each pellet was resuspended in 150ul of TE buffer(10mM Tris-HCl, pH 8.0; 0.1 EDTA). 300ul of SDS(1% SDS, 0.2 N NaOH) was added to each pellet. The tubes were placed on ice for five minutes, after which, 225ul of ice-cold 3M potassium acetate(pH 4.8) was added. The tubes were again placed on ice for five minutes and subsequently microfuged for five minutes. The supernatants were recovered and transferred to new tubes. One volume of phenol/chloroform was added to each new tube. The tubes were shaken vigorously for two minutes and centrifuged for five minutes. The upper, aqueous phase was recovered and transferred to a new tube. One volume of chloroform was added to each tube. The tubes were vigorously mixed and microfuged for three minutes.

Conflicts in the Godfather Essay

The Godfather is an insightful sociological study of violence, power, honor and obligation, corruption, justice and crime in America. Part I of The Godfather Trilogy centers on the Corleone crime â€Å"family† in the boroughs of New York City in the mid 1940s, dominated at first by the aging godfather/patriarch â€Å"Don† Vito Corleone. As a turn-of-the-century Silician immigrant, he is the head of one of the five Italian-American â€Å"families† that operates a crime syndicate. The ‘honorable’ crime â€Å"family,† working outside the system due to exclusion by social prejudice, serves as a metaphor for the way business (the pursuit of the American dream) is conducted in capitalistic, profit-making corporations and governmental circles. Although conflict in life may seem unpleasant, literature readers find it to be quite exciting. The characters involved within a conflict clearly stand out as to what they truly believe in and who they are as a person. In Mario Puzo’s, â€Å"The Godfather†, a number of conflicts are revealed through the Mafia underworld. The first, and most important conflict that is demonstrated in â€Å"The Godfather†, is revealed right away through â€Å"The Assassination Attempt on Don Corleone†. The Don shows the conflict of man vs. society by first refusing to enter the drug business. This decision, will ultimately effect the future conflicts that are revealed throughout the story. By refusing to participate in the drug business, the Corleone Family becomes outcasts of the five major crime families in New York. By doing so, the Don sparks the war between his family and the other five families. This war ends up lasting for several years and costs many lives. The second conflict that is present in â€Å"The Godfather†, is the conflict of divergent ideas vs. his father’s ideas. Santino, â€Å"Sonny†, Corleone, demostrates this conflict by deciding to go against his father’s beliefs. Sonny feels that drugs are the thing of the future and he decides that if his father dies, he will make the deal of entering the narcotics business. Tom Hagen, the adopted son of Vito Corleone, also believes that by entering the drug business, the Family will become much more successful later down  the road. A third conflict that is present in â€Å"The Godfather†, is the conflict of man vs. himself. This conflict is demonstrates through the character of Michael Corleone, the youngest son of the Don. Michael’s conscience is at war with his with his desire. After the assassination attempt on his father, Michael feels that he needs to get involved and help his father continue the Family business. However, his mind also feels that it is wrong to enter the criminal business. In conclusion, it is clear that many conflicts exist in Mario Puzo’s, â€Å"The Godfather†. They include, man vs. society, divergent ideas vs. father’s, and man vs. himself. Together, these conflicts create an intriguing plot that captures the viewer’s attention to the Mafia Underworld during the 1940’s and 1950’s in New York. Mario Puzo’s materpiece studies the power of conflicts that involve violence, power, honor, obligation, justice, and corruption.

Weeks v. United States The Case and Its Impact

Weeks v. U.S. was a landmark case that laid the basis for the exclusionary rule, which prevents illegally obtained evidence from being used in federal court. In its decision, the court unanimously upheld Fourth Amendment protections against unwarranted searches and seizures. Fast Facts: Weeks v. United States Case Argued: Dec 2—3, 1913Decision Issued:  February 24, 1914Petitioner:  Fremont WeeksRespondent:  United StatesKey Questions: Could the items obtained without a search warrant from Mr. Week’s private residence be used as evidence against him, or was the search and seizure without a warrant a violation of the Fourth Amendment?Unanimous Decision: Justices White, McKenna, Holmes, Day, Lurton, Hughes, Van Devanter, Lamar, and PitneyRuling: The Court held that the seizure of items from Weeks residence directly violated his constitutional rights, and also that the governments refusal to return his possessions violated the Fourth Amendment. Facts of the Case In 1911, Fremont Weeks was suspected of transporting lottery tickets via mail, an offense against the Criminal Code. Officers in Kansas City, Missouri, arrested Weeks at his work and searched his office. Later, officers also searched Weeks home, seizing evidence including papers, envelopes, and letters. Weeks was not present for the search and officers did not have a warrant. The evidence was turned over to the U.S. Marshalls. Based on that evidence, the Marshalls conducted a follow-up search and seized additional documents. Prior to the court date, Weeks’ attorney petitioned the court to return the evidence and to prevent the district attorney from using it in court. The court denied this petition and Weeks was convicted. Week’s attorney appealed the conviction on the basis that the court had violated his Fourth Amendment protection against illegal searches and seizures by conducting an unwarranted search and by using the product of that search in court. Constitutional Issues The main constitutional issues argued in Weeks v. U.S. were:Whether it is legal for a federal agent to conduct an unwarranted search and seizure of a person’s home, andIf this illegally obtained evidence can be used against someone in court. The Arguments Weeks’ attorney argued that officers had violated Weeks’ Fourth Amendment protections against unreasonable searches and seizures when they entered his home without a warrant to obtain evidence. They also argued that allowing illegally obtained evidence to be used in court defeats the purpose of the Fourth Amendment. On behalf of the government, attorneys argued that the arrest was based on sufficient probable cause. The evidence uncovered in the search served to confirm what the officers had suspected: Weeks was guilty and the evidence proved that. Therefore, the attorneys reasoned, it should eligible to be used in court. Majority Opinion In a decision delivered by Justice William Day on February 24, 1914, the court ruled that the search and seizure of evidence in Weeks home violated his Fourth Amendment right. Fourth Amendment protections apply to someone whether accused of crime or not,† according to the Court. Officers needed a warrant or consent to search Weeks home. The federal government also violated Weeks Fourth Amendment protections when the court refused to return evidence seized during an unreasonable search. In finding that the search was illegal, the court rejected one of the governments main arguments. The governments attorneys had attempted to show the similarities between Adams v. New York and Weeks case. In Adams v. New York, the court ruled that evidence incidentally seized while conducting a legal, warranted search may be used in court. Since officers had not used a warrant to search Weeks home, the court refused to apply the ruling reached in Adams v. New York. The Justices ruled that the illegally seized evidence was fruit from the poisonous tree. It could not be used in a federal court. Allowing the district attorney to use such evidence to convict Weeks would violate the intent of the Fourth Amendment. In the majority opinion, Justice Day wrote: The effect of the Fourth Amendment is to put the courts of the United States and Federal officials, in the exercise of their power and authority, under limitations and restraints as to the exercise of such power and authority, and to forever secure the people, their persons, houses, papers, and effects, against all unreasonable searches and seizures under the guise of law. The Court reasoned that allowing submission of illegally obtained evidence actually encouraged officers to violate the Fourth Amendment. In order to deter violations, the court applied the exclusionary rule. Under this rule, federal officers who conducted unreasonable, unwarranted searches could not use the evidence they found in court. The Impact Prior to Weeks v. U.S., federal officers were not punished for violating the Fourth Amendment in pursuit of evidence. Weeks v. U.S. gave the courts a means of preventing unwarranted intrusions on a person’s private property. If illegally obtained evidence could not be used in court, there was no reason for officers to conduct illegal searches. The exclusionary rule in Weeks only applied to federal officers, which meant that illegally obtained evidence couldn’t be used in federal courts. The case did nothing to protect Fourth Amendment rights in state courts. Between Weeks v. U.S. and Mapp v. Ohio, it was commonplace for state officers, unbound by the exclusionary rule, to conduct illegal searches and seizures and hand the evidence to federal officers. In 1960, Elkins v. U.S. closed that gap when the court ruled that the transfer of illegally obtained evidence violated the Fourth Amendment. Weeks v. U.S. also laid the groundwork for Mapp v. Ohio in 1961, which extended the exclusionary rule to apply to state courts. The rule is now considered a fundamental element of Fourth Amendment law, providing the subjects of unreasonable searches and seizures a unified manner of recourse. Weeks v. U.S. Key Takeaways In 1914 the court ruled unanimously that evidence obtained through an illegal search and seizure could not be used in federal courts.The ruling established the exclusionary rule, which prevents the court from using evidence that officers uncover during an illegal search and seizure.The exclusionary rule only applied to federal officers until Mapp v. Ohio in 1961. Sources Root, Damon. Why Courts Reject Illegally Obtained Evidence.  Reason, Apr. 2018, p. 14.  General OneFile.http://link.galegroup.com/apps/doc/A531978570/ITOF?umlin_m_brandeissidITOFxidd41004ce.Weeks v. United States, 232 U.S. 383 (1914).