Elucidating the p53 Signalling Network by Reverse Genetics

Elucidating the p53 Signalling Network by Reverse Genetics Reiyyan Tariq Nizami Aim In my research project I am working with four different strains Caenorhabditis elegans. I have been performing a double mutant screen using an RNAi knockout library. Cep-1 is a protein that causes apoptosis due to DNA damage in the worm. The cep-1 protein is an ortholog of the human tumor suppressor protein p53, which is found mutated in many cancerous cells. The aim of my experiment is to find genes that are regulators of cep-1 which cause lethality if mutated with cep-1. Alternatively mutants which are lethal as single mutants but survive as double mutants in combination with a cep-1 mutant are also potential genes of interest. These mutants have the potential to be regulated by drugs/proteins to change the levels of cep-1 and induce/prevent apoptosis in cells. Background Cancer is a disease of uncontrolled cell growth in our body. Under normal conditions a cell has regulatory proteins and check points that make sure that a cell is growing at the right pace and if for any reason the cell starts to grow irregularly then these proteins stop the cell growth and cause apoptosis. Apoptosis is the programmed death of a cell, as you can imagine it is tightly regulated and loss of regulation can result in catastrophe for the cell and the organism. When apoptosis is over active, healthy cells will die and this is often seen in diseases such as neurodegenerative diseases, hematologic diseases and many other tissue damage diseases. Loss of apoptosis in turn results in cancers, autoimmune diseases and inflammatory diseases. P53 is a tumor suppressor protein that was discovered in 1979. Since then a lot of research has been done into p53 and its functions. P53 is a tumor suppressor protein that is involved in defense of the cell. It is activated when a cell undergoes many various kinds of stress, such as DNA damage, hypoxia, metabolic stress and oncogene activation. It is one of the most important barriers to cancer in many mammals. P53 works in many various pathways, primarily p53 is involved in binding to transcription factors that then activate pathways involved in cellular defense, such as preventing angiogenesis and cellular growth. Malfunctioning p53 is one of the greatest hallmarks of cancer. The majority of mutations found in p53 are due to single nucleotide substitutions in the amino acids of the DNA binding domain of the protein. Similar cancerous phenotypes are observed when either p53 loses function due to a loss of function mutation or when negative regulators of p53 are overactive or positive regulators of p53 lose their function. However p53 is quite unique amongst tumor suppressor proteins because different missense mutations in the protein can cause different levels of p53 activity and hence have varying effects on the host. The effect of the mutation is further modified depending on the genetic background of the person with the mutation. Due to the different effects of various amino acid substitutions and the varying genetic background of patients it is quite difficult to study large populations. This is because high-throughput sequencing and genome wide single nucleotide polymorphism maps are expensive to obtain. The prices are going down as new technologies are becoming available however until now there have been no larger scale studies relating different mutations to varying levels of risks for different types of cancers. It is also hard to perform molecular studies on entire humans and so research is done on cell lines, mice and on cep-1 the Caenorhabditis elegans ortholog of p53. Caenorhabditis elegans is a worm in the nematodes phylum. They are a very well-studied organism due to their ease of study. C. Elegans are one of the simplest organisms that have a nervous system and that make them a very good model organism for neuronal studies. They are transparent creatures and so many molecular and cellular processes especially those involved in development have been thoroughly researched. They have a short generation time and are very cheap to maintain. They are an extremely good model organism for performing screens because any of their nonessential genes can be knocked out easily by using RNAi. The cep-1/p53 pathway is highly conserved throughout evolution as it is an extremely important pathway that is essential for cellular survival over time. The cep-1 protein works through the following pathway to cause apoptosis in cells suffering DNA damage. Cep-1 à ¯Ã†â€™Ã‚   Egl-1/Ced-13 –| Ced-9 –| Ced-4 à ¯Ã†â€™Ã‚   Ced-3 à ¯Ã†â€™Ã‚   Apoptosis A similar pathway is observed in humans p53 à ¯Ã†â€™Ã‚   BH3 –| Bcl2 –| Apaf1 à ¯Ã†â€™Ã‚   Caspase à ¯Ã†â€™Ã‚   Apoptosis A majority of these proteins are orthologs to one another. It was hoped that through my screen further proteins which interact with Cep-1 would be found and then their orthologs in humans could be found and then researched and targeted to regulate p53. Materials and Methods The experiment in itself was an extremely simple but time consuming experiment. Screens were performed with mutant worm strains which were then fed RNAi, through Escherichia coli (E. coli), to silence the gene of interest and create double mutants. To begin with a liquid screen is preferred over a solid screen. This is because with a liquid screen you can screen a larger sample of double mutants more easily than a solid screen. Liquid Screen The following strains of bacteria and worms were utilized, E. Coli – OP50, C. Elegans – N2, C. Elegans – GK138, C. Elegans – LG12501. E. Coli – OP50: Food source for C. Elegans C. Elegans – N2: Wild type worms C. Elegans – GK138: Cep-1 Mutant worms C. Elegans – LG12501: Cep-1 Mutant worms RNAi Knockout library for C. Elegans Chromosome 1 Day 1: Grow worms on Nematode Growth Medium (NGM) plates with OP50 as a food source for the worms. Day 3: Bleach worms that were plated on day 1 so as to only have eggs remaining on plates. Bend glass rod into L shape using a high temperature flame (Bunsen burner) Pipette bleach onto plates with worms on them and gently scrape the surface to mix worms and eggs with bleach Pipette fluid into Eppendorf tube and centrifuge at max speed for 3 minutes Aspirate most of the fluid and keep as much of the pellet as possible Suspend pellet with bleach Repeat steps 3 and 4 Suspend pellet using M9 solution Centrifuge at high speed for 1 minute Aspirate most of the fluid and keep as much of the pellet as possible Repeat 7 and 8 Suspend pellet using M9 solution and vortex Eppendorf tube at low speed to mix solution Place Eppendorf tubes in slow rocker in a 20 ° fridge overnight to allow eggs to hatch Day 4: Replicate bacteria containing RNAi from knockout library using a sterile 96 pin replication tool into a 96 well containing Liquid Broth (LB) with Ampicillin and allow the bacteria to replicate overnight at 37 °C in an incubator Induce transcription of RNAi using adding 0.1 Molar IPTG into wells and place in a shaker for 1 hour. Pellet bacteria by centrifuging in a cold centrifuge at 5 °C for 5 minutes at 2,500 g Remove the supernatant by flipping over the well quickly but carefully so as to keep pelleted bacteria in the wells Suspend bacterial pellet in wells using NGM Pipette worms into wells and place in 37 °C shaker Day 8: Remove worms from shaker gently making sure not to tilt the 96 well plates Observe and record phenotype of the worms Compare phenotype between the 3 different strains of worms Leave worms in 20 °C fridge overnight Day 9: Remove worms from fridge gently making sure not to tilt the 96 well plates Observe and record phenotype of the worms Compare phenotype from previous day Compare phenotype between the 3 different strains of worms Sequencing After potential hits were found in the liquid screen the RNAi from these bacteria were sequenced to ensure that the sequence of the RNAi was correct and hadn’t randomly mutated over time. RNAi was prepared by using a Qiagen miniprep spin kit. Suspend bacteria in 250  µl Buffer P1 and place in a microcentrifuge tube Mix 250  µl Buffer P2 and shake the mixture by flipping tube over a few times To the mixture add 350  µl N3 buffer, mix well quickly Centrifuge at 13000 rpm for ten minutes Aspirate supernatant into new tube Centrifuge again for a roughly 1 minute and discard the flow through Wash spin column with 0.5 ml PB buffer and centrifuge for 1 minute, discard flow through Wash spin column with 0.75 ml PE buffer and centrifuge for 1 minute Discard flow through and centrifuge at maximum speed for 1 minute Place prep column in a sterilized 1.5 ml microcentrifuge tube Add 50 ÃŽ ¼l of water to prep spin column and allow to rest for 1 minute after which centrifuge for 1 minute After the Qiagen miniprep is complete the tubes were sent for sequencing to The Centre for Applied Genomics where it was sequenced and results were obtained within a week. Solid Screen Solid screens were performed on genes which were found to have increased lethality with cep-1 deletion or increased survivability with cep-1 deletion. The following strains of bacteria and worms were utilized, E. coli – OP50, C. Elegans – N2, C. Elegans – TG12 E. Coli – OP50: Food source for C. Elegans C. Elegans – N2: Wild type worms C. Elegans – TG12: Cep-1 Fluorescent tagged (GFP) worms C. Elegans – Ned-8: Positive control C. Elegans – HT115: Negative control RNAi Knockout library for C. Elegans Chromosome 1 Day 1) Streak RNAi bacteria of interest from RNAi Library to obtain single colonies. Allow them to grow overnight in 37 °C incubator Day 2) Pick and grow a single colony in 5 ml of LB + Amp + Tet overnight in a 37 °C shaker Day 3) Add 0.1M IPTG for 4 hours to induce RNAi Plate 100  µl of induced bacteria on RNAi plates incubate at 37 °C incubator overnight Day 4) Pick 5 worms at the same stage and plate onto RNAi plates Allow to grow over 4 days at 20 °C Day 8) Score phenotypes and compared between different strains Day 9) Score phenotypes again on next day and compare between different strains and the previous day Fluorescent Microscopy We used a Differential interference contrast (DIC) microscopy to observe localization of cep-1 in TG12 worms. Worms were mounted using the following method Place a drop or two of hot liquid agarose onto microscope slide Immediately place a second slide perpendicularly on top of the agarose Allow agarose to settle for one to three minutes Gently slide off second slide so as to leave an agarose patch behind Add a drop of 1mM Levamisole to paralyze worms and prevent their movement Pick worms and place them on the slide Slowly place coverslip on top of worms, be very gentle Once prepared the slides were observed using a DIC microscope to find any irregularities in the localization or amount of cep-1 in the worms, especially in the germline and the eggs. Results The aim of this experiment was to find genes that cause lethality as double mutants with cep-1 mutants but not in wild type worms, these genes would be positive regulators of cep-1 and cause apoptosis through cep-1. The screen was also designed to find worms that cause lethality in wild type worms but not in cep-1 mutants, these would be negative regulators of cep-1 and cause apoptosis through cep-1. The results were gathered and tabulated to allow for an easier and better analysis of data. Note: There is a lot of data and so only data that is relevant is shown. Liquid Screen Results Legend L = Synthetic LethalE = Embryonic LethalG = Slow GrowthB = Egg laying abnormal R = No RNAi C = Contamination A = Larval Arrest S = Sterile V = Variable Morphology P = Lethal progeny W = No Worms Cells of interest are highlighted Chromosome 1 – Plate 1 N2 1 3 5 7 9 11 13 15 17 19 21 23 A R G G/L L C A/C A/C E R E/S G R R I S/L S R R R R K R R R R M R R R R S O R R S/E R cep-1 (gk) 1 3 5 7 9 11 13 15 17 19 21 23 A R L C/L C/L G/E C G/S G/L C/L E R E/S G R R I E/S E/S R R R R K R R R R M R G/E R R R /G/S O R R S R cep-1 (lg) 1 3 5 7 9 11 13 15 17 19 21 23 A R C/L L L E/P C G/L G L E R E/S G R R I S/B S/B R R R R K R R R R M R E/G R R R S O R R S/E R Chromosome 1 – Plate 5 N2 2 4 6 8 10 12 14 16 18 20 22 24 A C E G E I E K R M R R R O G R cep-1 (gk) 2 4 6 8 10 12 14 16 18 20 22 24 A C E G I E K R M R R R O R cep-1 (lg) 2 4 6 8 10 12 14 16 18 20 22 24 A C E G I E K R M R R R O R Key results of interest in liquid screen Lethality in gk and lg but not in N2 Well Gene N2 gk lg A23 F53G12.5 G/E E/P M03 Y95B8A_85.h G/E E/G Lethality in N2 but not in gk or lg Well Gene N2 gk lg G04 F25H2.10 E Solid Screen Results 3 hits of interest were obtained from all the plates that were screened. These 3 hits were then repeated multiple times on solid media to make sure the results were accurate. N2 lg gk A23 -1 A23 -2 A23 -3 N2 lg gk M03 -1 E M03 -2 E M03 -3 N2 lg gk G04 -1 L A G04 -2 E G04 -3 The M03 strain showed opposite results in the solid screen in comparison to the liquid screen. However since the results were consistent we decided to study both M03 and G04 under the microscope with fluorescent markers. Fluorescent microscopy results No major mislocalization of GFP tagged cep-1 was found and no extra or lack of GFP tagged cep-1 was observe

Growth and Development of Tourism in Australia since World War II Essay

This is a short but informative discourse on the growth and development of tourism in Australia since World War II. The first few segments will focus on the history of Australia as a nation in order to provide a solid background for the inherent factors which contribute to the growth and development of tourism in Australia. The second segment will briefly highlight the Australian economy to show the benefits that have been derived from the tourism boom in Australia. Finally, the last segment shall carefully identify and explain the major factors that have contributed to this tourism surge in Australia since World War II. Brief History of Australia Australia mainly known for its picturesque surroundings is more than meets the eye. Australia boasts of a rich history under the rule of the British and enviable living conditions for its citizens (Pike, 2002). However, Australia was not always like this. Before the arrival of Western influences, Indigenous Australians inhabited the mainland but eventually European explorers set foot on Australia. The most notable explorer, James Cook mapped out the east coast of Australia and claimed this piece of Australia as a British Colony. The contributions of James Cook were integral in the establishment of a penal colony in New South Wales. The British territories grew larger with more and more discoveries of Australia’s mainland. The influx of the British caused the decline in the population of the Indigenous Australians (Pike, 2002). Slowly, the British colonies in Australia governed themselves while remaining under the British Empire. The British Empire only controlled foreign affairs, international shipping, and defense. A decade of planning, consultation, and voting gave rise to the Commonwealth of Australia on January 1, 1905 (Mason, 2003). The new country took advantage of all its natural resources to rapidly develop into an agricultural and manufacturing country taking into consideration its strategic location. World War I started to brew across the Europe and Australia was willing to take part in the war (Pike 2002). Enlistments were abundant which caused the industries and trade agreements of Australia were disrupted because of the war. Trade agreements with Germany were put to a halt thus Australia had to find other means to address their needs of steel. Like in any war, profiteering occurred and the ordinary people could not even purchase the basic utilities (Mason, 2003). However, at the end of the War, Australia managed to produce more products than ever before, which contributed to the growth of the nation. Effect of World War II on Australia World War II made Australia purchase products which could have easily been produced thus the war was a huge boon to Australia. The restrictions of rationing imposed by the government allowed personal savings to increase. There was also full employment during the war since manpower was being utilized for essential industries. Australia was more prepared for World War II than it was for World War I. World War II did affect Australia. â€Å"Migration revolution† began which led to multiculturalism instead of â€Å"White Australia†. The government was so afraid to be invaded that it encouraged the influx of refugees and displaced families from Europe. Consequently, Australia evolved into a country of mixed races and mixed cultures (Pike, 2002). Ties between Australia and the British Empire were officially severed with the passing of Australia Act 1986. This act severed any rule of the British Empire in the Australian states. Australia now rules itself and is developed country surrounded by the developing countries of the Asia-Pacific region (Mason, 2003). The Australian Economy Australia is a very prosperous nation. It boasts of ranking third in the UN Human Development Index 2006, which indicates that Australia is a developed country. It beat the United States and even the United Kingdom. Australia was only surpassed by Norway and Iceland. Australia developed from a country with an industry focusing on wool to a country that has numerous industries in sectors ranging from trade to agriculture (Macfarlane, 1998). In 1850, Australia focused on the wool industry but with the discovery of gold in 1850-1860, the government passed a bill that encouraged the manufacturing and construction. Since the gold resources were being stressed by the immigration, wool resumed its position as the economic power in 1860. The decade between 1880 and 1890 saw a great rise in foreign investment but foreign investors grew concerned about the returns that Australia could supply thus, Australia experienced a great depression by 1890. The world wars encouraged Australia to produce more products thus sectors like agriculture increases (Parham, 2002). Now, wool was not the only contributor in the economic strength of Australia. An injection of foreign investment allowed the development of Australia also contributing to the economic growth of the country. The beginning of World War II marked the period of increased productivity or the â€Å"long boom† (Parham, 2002). Manufacturing became a huge part of the economy. Import restrictions allowed the manufacturing sector to grow. However, foreign investors noticed the manufacturing sector could not increase productivity and investment declined in this sector. This did not affect the growth of the economy since the mining industry to exploit the natural resources of the country was being established and encouraged an increase in foreign investment. The Australian Stock Exchange was established in 1987. Government run industries like Commonwealth Serum Laboratories and Quantas were privatized. The Labor markets were deregulated and reforms like the Goods and Services Tax were implemented. All these contribute to a solid economic status that Australia enjoys (Burns, 2006). Tourism in Australia As shown in the graph presented below (See GRAPH 1), the revenues that Australia earns from tourism accounts for a significant portion of its GDP. In line with this, the Australian government has slowly developed the growth of the Tourism industry in the country. Some of these plans include the development of infrastructure to foster sustainable tourism and the shift to more ecotourism which accounts for a major part of Australian tourism (Edgel, 2006). Since World War II, the Australian government has been trying to encourage the growth of the tourism industry by taking advantage of the vast natural resources and exotic scenery. At the present, the tourism industry accounts for 4. 7% of Australia’s GDP. It is also the reason for 11. 2% of Australia’s export earnings and the employment 6% of the workforce in Australia (Elliot, 1997). The vast expanse that is home to a significant number of unique flora and fauna also accounts for the growth of the tourism industry in Australia. Major Factors Contributing to Tourism Surge in Australia Perhaps the greatest factor contributing to the growth of the Australian Tourism industry since the World War II was a work ethic that was long in place, even before the occurrence of World War II. In the early 1900’s, the Australian Urban Trade Unions adopted an eight (8) hour work day accompanied by eight (8) hours of rest and most importantly, eight (8) hours of recreation (Burns, 2006). This was an important development in the growth of local tourism and even international tourism because it emphasized the relevance of leisure and recreation and made travel and tourism an integral part of Australian culture. This type of culture and thinking prompted the creation of new tourist destinations for the local residents. This led to the development of railroads which in turn led to greater access to most of Australia’s national parks (Herremanns, 2006). The developed that ensued not only made tourism accessible to the urban dwellers but eventually opened up tourism to all sectors of society. The development of coastal parks began soon after this. Inland and outland tourist destinations also soon flourished because of the development of the infrastructure in Australia (Chon, 2000). Nina Mistilis, in her work entitled Public Infrastructure Development for Tourism in Australia A Critical Issue (2004), is of the opinion that the past and present growth of the tourism industry in Australia is largely dependent on the infrastructure projects undertaken by the Australian government. For tourism to continue its growth, public infrastructure projects such as roads and rails, ports and airports must be developed to accommodate more traffic and also allow better access to other areas in Australia. Another major factor in the growth and development of tourism in Australia was the amount of leisure time and greater disposable income as a result of the Second World War (Elliot, 1997). The first change was the gradual institution of three (3) weeks holiday as a standard. This allowed more of the domestic tourists to enjoy their vacations in Australia and visit places that were farther away from their hometowns thus expanding the tourist sites (Herremanns, 2006). Aside from the increased holiday and leisure period after the war, more and more Australians owned motor vehicles due to the greater disposable income that was available. This meant that the three (3) weeks holidays could be enjoyed more as the availability of alternative modes of transportation permitted tourists to explore other tourist destinations all throughout the continent (Mason, 2003). International tourism, which accounts for a significant portion of the tourism industry of Australia, also contributed to the development of tourism after the Second World War as it made Australia a viable place to travel to (Burns, 2006). In the early 1980’s and 1990’s when Japan and America had turned into powerhouse economies that possessed strong currencies, particularly against the Australian dollar, international tourism skyrocketed (Herremanns, 2006). The profitable short package tours, which accounted for majority of the tourism packages sold, featured trips to the iconic symbols of Australia such as the Koala Bear, Ayer’s rock and the Great Barrier Reef (Herremanns, 2006). These short package tours were particularly significant however as they increased the marketability of Australian tourism and also strengthened the association of those symbols with Australia and thus increasing visibility and identity on a global scale (Mason, 2003). The local economy invariably profited from these also as the short package tours generated a lot of income and also increased souvenir item sales. This economic growth increased the per capita earnings of the population in these local attractions and also improved the overall condition of the tourist attractions as the added source of funding and income allowed improvements to be made to the local infrastructure (Mistilis, 2004). Another important phase in the growth and development of tourism in Australia since World War II was the development of ecotourism (Edgel, 2006). The vast natural resources and natural land attractions of Australia, not to mention the rich and unique flora and fauna that abound in the national parks, make it an ideal place for ecotourism (Pike, 2002). Backpaking soon became a major source of revenue for the local tourist destinations. Because ecotourism promotes the overall growth of the community, long term benefits were provided to the local populace to achieve what is known as â€Å"sustainable tourism† (Edgel, 2006). This also allowed for the preservation of the local cultures that make Australia and unique and enjoyable tourist experience, be if as part of a group or alone.

Book Review of A National Party No More

The Conscience of a Conservative Democrat by Zell Miller In A National Party No More Senator Zell Miller writes a non-fiction book that is something of a memoir of his political life as a lifelong Democrat and as well as being a diatribe against the Democratic Party. In 2002 the Democratic Senator Paul Coverdell from Georgia died suddenly and the Georgia Governor Roy Barnes asked Zell Miller to fill in until November of that year and then to run for the position to serve the time remaining in the late senator's term of office. When Miller went to Washington D.C. he claims that he had hoped that he would find Washington to be â€Å"the place where great issues of the day are debated and solved, and great giants walk those hallowed halls.† Instead he discovered what Washington D.C. was not at all like he had hoped and this angered him â€Å"on behalf of Americans† (Miller 8). In his career Miller has served the State of Georgia as an administrator of a number of â€Å"vital agencies, as an assistant to two governors, as head of the State Democratic Party, as Lieutenant Governor, and then as Governor† (Miller introduction no page number). He also served in the Georgia State Senate from 1965-1969. He failed in his attempts to run for the U.S. House of Representatives from Georgia. In addition Miller served in the U.S. Marine Corps and has taught at four different colleges. It is worth noting that the majority of these positions are executive positions not legislative positions so serving as Georgia's United States Senator put him into a relatively unknown form of government where he lacked extensive experience and none on the national level. It appears somewhat incongruous that a marine would be a lifelong Democrat since military personnel currently tend to lean toward the Republican Party. Although Miller served only three years in the Corps it is clear that his experiences affected him greatly because he has written a book Corp Values: Everything You Need to Know I Learned in the Marines.† Miller's book was occasioned by his experiences in the U.S. Senate beginning in 2000, in combination with his advanced age. Miller was born in 1932 according the reverse side of the title page. The book was published in 2003 so Miller was 70 or 71 at the time his book was published. This is certainly not an issue of itself, but one wonders if Miller would make the same claims he had entered the same Senate at the age of forty or fifty. Miller alludes to this when he writes that he has â€Å"arrived at a station in life where I hear the whistle of that moral policeman we all have to answer to . . .† (Miller 1). He takes the occasion to advise â€Å"members of my Democratic Party and other politicians who are so far out of touch with regular Americans to ‘shape up'† (Miller 2). This position, that tacitly assumes he is correct while other party members who disagree with him are wrong, is strongly reminiscent of an elderly gentleman on the brink of retirement who climbs on a soapbox to â€Å"straighten out† the next generation. This is not to say Miller does not make some good points, he just does not prove them nor give the reader sufficient information to determine if Mill is correct. The book suffers significantly because it has no notes, no bibliography and no index. Consequently the reader has no opportunity to check either the statements Miller makes as being either true or false. Many of his claims are supported by anecdotal evidence based on his memory of what happened throughout his career. Miller calls himself a Conservative Democrat, an unusual designation, but not an illogical one. Despite this he is known to have been a supporter of President George W. Bush and announced in 2003 that he would support the Presidents re-election. He spoke at the Republican National Convention in 2004 in support of the President. Such actions hardly indicate a lifelong Democrat. Miller criticizes the Democrat Party because the leaders, he claims, have ignored the opinions of Conservative Democrats in the South, about one-third of the U.S. population, and have told them to â€Å"go to hell† (Miller 9). He appears to assume that all Democrats in the South are Conservative Democrats because he is one and that they all agree with him. He fails to mention the non-conservative Democrats in the South and seems to assume there are none. Miller concludes the Democrat leadership disagrees with Southern Democrats on the critical issues of â€Å"capital punishment, late-term abortion (even with a lot of pro-choice people), trying juveniles as adults, national defense, and the teaching of values in school† (Miller 3). Miller's reasoning process is suspect for a variety of reasons. First, it is not clear that these are the critical issues, at least on a national level. Capital punishment has been left up to the states and should not be regarded as a national issue, as are abortion laws as long as laws do not restrict a woman's right to control of her own body. Trying of juveniles as an adult does not seem to be a national issue either nor should it be. The current system presumes juveniles will not be tried as adults unless there are significant overriding reasons for doing so. The decision of where to try a juvenile is judged on a case-by-case basis which is as it should be. As far as the teaching of values as a national issue, it is clearly an important issue that impacts people throughout the country, but the Federal Government has no basis to determine what values should be taught. Miller seems to have completely forgotten the Tenth Amendment to the Constitution that reserves to the states any powers not delegated to the federal government and not prohibited to the states shall be a state power. It is apparent that many of these critical issues are state issues. National defense clearly is a national issue, but federal taxes, the deficit, Medicare, and Social Security benefits are as well, but Miller doesn't mention these issues. Miller's has a rambling and folksy writing style that slips from story to story and slides from point to point in a chaotic fashion that defies linear analysis. He chooses his anecdotes by cherry picking stories that will reinforce his position even if they are not related to the Democratic Party; on page 145 Miller quotes The Rubaiyat of Omar Khayyam, Winston Churchill, and Rodney King on the same page in support of a chapter entitled â€Å"Give to Bigotry No Sanction.† Certainly a case can be made for citing Rodney King, but the other two sources were clearly not written in support of civil rights in the United States. To his credit Miller admits mistakes he has made during his career. He points out that during 1964 he had â€Å"proclaimed that there should be an ‘investigation of Communist infiltration in the civil rights movement.' What an idiot!† (Miller 143). This is remarkably refreshing given the current climate of politicians who consistently hide what they have done and deny they have done it until they are proven to have done it when they will admit and ask for forgiveness. On the other hand this begs the question as to just how much credence one ought to place in the writings of a self-proclaimed idiot. Miller claims that the Democratic Party no longer represents the majority of Americans and has become distinctly too liberal in relation to the United States' population as a whole. This is an interesting position. Miller does not claim the leadership is wrong on issues, just that they disagree with Southern Conservative Democrats such as himself. This leaves the possibility open that the leadership is correct and the membership is wrong, but Miller fails to allow for this possibility. If this proves to be the case, it appears that Miller is advocating that the Democratic Party concern itself, not with the correct solution, but with gaining power again. A National Party No More: The Conscience of a Conservative Democrat is largely not successful. His conclusion has merit but his treatment of the issues is inferior. The lack of references is a major weakness that could be easily corrected. Miller's failure to move in a linear, logical fashion in favor of using a disjointed, episodic style greatly reduces the effectiveness of Miller's writing. Rather than being the scathing indictment he hopes to provide that will help the Democratic Party, Miller's book feels more like a farewell by a statesman whose party has evolved while he has not. His gloom and doom predictions for the Democratic Party made in 2004 proved him incorrect since the Democratic once again gained the majority in both houses. Although Miller makes some interesting points that have validity, his book should be read with circumspection. Works Cited Barnes, Fred. â€Å"Zell Miller Endorses Bush.† 29 Nov. 2003. The Daily Standard. 20 April 2007. Miller, Zell. A National Party No More: The Conscience of a Conservative Democrat. Atlanta: Stroud and Hall Publishing, 2003. â€Å"Text Of Zell Miller's RNC Speech.†Ã‚   01 September 2004. CBS News. 20 April 2007   ;http://www.cbsnews.com/stories/2004/09/01/politics/ main640299.shtml;.

Effective Communication Within The Workplace - 1254 Words

Introduction In life, individuals who work often find that in order to operate effectively within a workplace their must be effective communication between staff and managers from front line managers to high end managers. However, in workplaces all around the world. Employees often fail to address employees effectively and often as a direct result. Employees either make mistakes or purposely choose to disregard orders. To reinforce this statement managers still evaluate that there is a clear failure of effective communication within the workplace still is a large problem. Because of this communication in the workplace is now being seen as a cornerstone for effective productivity and a positive employee-employer relationship as well as for profit and non-profit organisations (Marques, 2010). As both a direct and indirect result issues arise from poor communication and can be detrimental to the success and productivity of an organisation. It is the role of management on all levels of the organisa tion to prevent this from occurring. Interpersonal is the way in which individuals converse information, feelings, and meaning through one or both verbal and non-verbal messages. Interpersonal communication is often perceived to be face to face contact. Where often it is generally presumed that this type of communication in fact is assumed to be spoken language however, it is not actually what is said, but in fact the way in which language is delivered and perceived by others. ThisShow MoreRelatedEffective Communication Within A Workplace854 Words   |  4 PagesCommunication skills are very important no matter where or who you are. Communication within a workplace can determine whether a business or individual is successful or not. Understanding effective communication in a medical setting helps hospitals and doctor offices develop a work environment that is able to communicate effectively with coworker, patients and doctors in order to take of the patients needs. Listening, clarity and Patience are three main keys to effective communication within aRead MoreEffective Communication Within The Workplace1848 Words   |  8 Pagesorganisational communication; it is the communication that happens within the organisational, whether it is face to face communication or video conferences communication. It is important to note that if organisations do not communicate the right information, in the right way, to the right people, at the right time, this could create problems and have an impact on the level of effectiveness and efficiency. In essence, there is therefore a need for effective communication within the workplace. If thereRead MoreInterpersonal Communication And The Workplace946 Words   |  4 PagesInterpersonal communication in the workplace is developed positively or negatively on the individual relationships we have combined with our human behaviors and human actions within each of those relationships. There are many things that can affect interpersonal communications within the workplace, from generational that create technological gaps, to diversity and tolerance it creates, and finally the type of workplace, is it a team environment encouraging inclusiveness within the organization orRead MoreEffective Communication Skills Are Essential For Success1222 Words   |  5 PagesIntroduction Effective Communication skills are essential for success in every organization. Individuals today in the workplace need to effectively communicate with employees, customers and potential clients. Effective and clear communication if not delivered in the right manner could be interpreted by the receiver and have a negative impact. There are many barriers that can effect a conversation such as cultural differences, gender and environment. Employees can learn how to adapt in differentRead More Work Environment Paper1015 Words   |  5 Pages Work Environment Paper Good communication skills are imperative for organizations to carry out in an attempt to maintain outstanding performance from employees and management. Every workplace is unique in terms of their diversity profile therefore, is it vital for organization to create and execute creative communication methodologies that will enable organizations to develop a community rather than just your average run of the mill work environment. For example, active listening, collective goalRead MoreEffective Communication in the Workplace Essay1220 Words   |  5 PagesCommunication Barriers in the Workplace Communication barriers in the workplace can have a serious effect on the functioning and of an organization. In the following article we shall understand what some of these communication barriers are and how to overcome them. Ads by Google Improve Communication Our NLP Training Program Helps You Overcome Your Fears. Enroll Today! www.EasyNLP.com/ Conflict Management How much is conflict costing you? Assessment, Training, Coaching www.StrategicLeadershipCoachingRead MoreImportance of Cross Cultural Communications1209 Words   |  5 PagesEffective cross-cultural communication is one of the most important issues dealt with in business, particularly when a firm operates at an international level. Communication is a process with three key elements, which includes a source, an audience, and a channel. Communication derived from businesses will have listeners that include, but are not limited to customers, employees, suppliers, and the community (Caddy et al.). These listeners contribute to the success of a firm, which is why there isRead MoreThe Effects Of Effective Communication On Job Performance1680 Words   |  7 PagesWhen asked to write a reflective think piece describing a single communication issue as it pertains to the organization in which I work, the Employment Development Department, I decided to delve into the to pic of effective communication, especially in regards to job expectations. I choose this particular aspect of communication because it is one of my divisions biggest complaints in regards to management or the lack there of, and I want to better understand how to avoid miscommunicating with fellowRead MoreThe Importance Of An Undergraduate Business Management Student1535 Words   |  7 Pagesever-changing business world, a key component to the productivity within the workplace is communication. As an undergraduate business management student, an important part of communication is understanding the difference between maintaining the faà §ade of an authoritative figure or maintaining a friendly relationship with employees and how it would affect the attitudes, behaviors, productivity and quality of work. In order create a successful workplace environment, finding and establishing a middle ground betweenRead MoreEssential Attributes in Nursing1220 Words   |  5 Pagescommunities. I n a nursing good workplace citizenship needs an understanding and maintaining a good relationship between patients and health professional to deliver quality health care and services. In the context of nursing, communication, teamwork and social responsibility is the main graduates attributes that are essential in workplace citizenship because it promotes a healthy relationship between health professionals, patients and improve the nursing workplace. Communication is the exchanging information