Good Mothers Bad Gang

Anita Anderson Dr. Mike Calvert Teenage Wasteland 18, July 2011 Good Mom/Bad Gang Kids on the street, parents that don’t care, looking for a sense of family or belonging; do you see a child at risk? While in some cases this may be true, in many instances this is not the case. In her article â€Å"Mothers, Sons, and The Gangs† Susan Horton tells of three different gang members and their families. Although the families had different backgrounds, beliefs, and techniques they all shared in common sons who were gang members. They are mothers who care but have to face alone the battle of raising their sons in a community lost to gangs. In Horton's article we are introduced to three different families whose sons are involved with gangs. Teresa Rodriguez who still feels foreign and timid about the culture of America. Maggie Garcia who was raised in the same neighborhood as her son and believes that loyalty to the neighborhood is just and understandable. Gayle Thomas Kary, because of financial struggles was thrown into the gang community and fought her best to save her son. These young men were all minors living in a society pressured by gang affiliation. Although their homes had loving mothers their gangs had friends, boldness, acceptance, and a way of life. â€Å"There may be several deep-seated, underling motives for joining gangs: â€Å"Those who join gangs are struggling with the classic, desperate needs of adolescence: for the end of childhood; for acceptance as an adult; for sexual, social and economic identity; for status and success; for respect and a sense of belonging†. Sheldon, Tracy, Brown 77) Not all of these aspects are apparent in gang members but at least one is. In Teresa Rodriguez's case her son at thirteen unbeknown to her belonged to the gang Sur 13. Because of his boldness he was shot and the family home has been victim to drive-bys ever since. The bullet missed his heart and fortunately he survived. Still he did not learn his lesson and claimed that he did not care if he died. Finally his mother threatened to send him to l ive in Mexico if he continued to go out. â€Å"He did not want that, so he stayed inside. But that has not stopped him from dressing or acting like a member of the Sur 13. After the incident of the shooting Rodriguez also found out that all three of her older sons were part of gangs. Her 16-year-old threw away his cholo clothes right when he heard about his brother. He had not been with the gang since then. The two older boys are very repentant, but it is the father had always left rearing the children to the mother he did not have a strong, active, male role model. So, perhaps he felt as if he had no other alternatives. Maggie Garcia also has a son who is a gang member but her view and approach to her son’s affiliation is very different. She was raised in the same neighborhood as she raised her son in; she understands how much the neighborhood could mean. During school her son got into a fight with a rival gang member because each claimed a different neighborhood. Her son was expelled and she felt the principal at his school was upset because my son had stated that he would die for his neighborhood, but if he had said that if he would have died for his country, the principal probably would have given him an award. This mom understood that the danger in the inter-neighborhood conflicts that her son gets into because other family members have been lost to such conflicts. Sadly this is not enough for her to forbid her son from being with his friends. She feels that by doings so would be like her son telling her not to be with her friends. Garcia follows similar problems as the Rodriguez family; she mentions her history and connection to the neighborhood. In this story the father is also absent and the mother supports her son and find it difficult to believe that her son is wrong. She does not have a bad relationship with her son but he receives no discipline or clear-cut parameters from her. She seems to feel that he is loyal to his friends and at times perhaps takes his loyalty a little too far. Garcia seems to adopt the â€Å"boys will be boys† attitude and does not want to believe that her son may be committing crimes or may be with the wrong crowd or may be in a gang. When he was arrested she insisted that he was just in the wrong place at the wrong time. She could not accept that maybe he is doing wrong and that he should have known better than to be in such situations. She tells him to stay out of the fire but her words are not strong enough to keep him out. Although she is not a bad mother, she is not giving her son reason to stay out of gangs. Because she could no longer afford to live in a community where her son had more opportunities and alternatives her son joined the only other thing, the gangs. The story of Gayle Thomas Kary may be the saddest one because the she tried everything and when she finally thought she succeeded her son, Jamee, died as a result of gang violence. Kary did more than the others, discipline, rules and action. When she caught her son cutting up soap to look like cocaine she sent him to his father, perhaps a change of venue would make the difference, but three weeks later, his dad sent him back because he couldn’t control him. She tried reasoning with him telling him â€Å"†¦ fast life goes fast. â€Å"(Horton) When he was arrested for possession of cocaine Kary was highly relieved but to her disappointment he was let out on probation. Kary refused to accept him and he was sentenced to juvenile hall and a youth camp. When he was released he had a probation officer Kary could turn to. Jamee was tired of life on the streets and his probation officer convinced him to turn himself into a youth facility away from Los Angeles. Finally things were turning around and he really seemed to have changed. He was like the child she use to know Unfortunately when she let him go out with his friends to get something to eat he was shot and died. She did everything she could but still she could not save her son's life. Unlike the other two mothers Kary was really able to turn her son Jamee around. She finally made him understand and she can say that his death was an accident. He was not just a number or a statistic of children lost to gang violence. Although, these mothers were, loving they could not help that a major factor was absent from their son’s lives, their fathers. The fathers may have been there but none seemed to take an active role in their children's lives. Parents can make a difference in their children's lives but many times they cannot do it alone. Gangs are not only the problem of the parents it is a problem of society. â€Å"To prevent gangs, society, police, and parents have to work together to keep gangs out of neighborhoods and also to address the existing gang problem†. Ojile) It may be easier to blame the parents but the truth is the parents are not only the ones to blame. We have to realize that in a society where gangs are the neighborhood it could be our son, our daughter, our nephew, etc. that is lost in the gangs. Works Cited Horton, Susan. â€Å"Mother's, Son's and the Gangs. † Los Angeles Times 16 Oct. 19 88. Print. Ojile, Todd. â€Å"Gang Activity in Wichita. † Teenage Wasteland. Baker University, Wichita Campus, Wichita, Kansas. 25 July 2011. Lecture. Shelden, Randall G. , Sharon K. Tracy, and William B. Brown. Youth Gangs in American Society. Belmont, CA: Thomson/Wadsworth, 2004. Print.

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Human Resource Management has evolved significantly over recent Assignment

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Energy Drink and Red Bull Essay Example for Free

Energy Drink and Red Bull Essay This global profile focuses on the industry trends in soft drinks. ? All values expressed in this report are retail/off-trade in US dollar terms using a fixed exchange rate (2012). ? 2012 figures are based on part-year estimates. ? All forecast data are expressed in constant terms; inflationary effects are discounted. Conversely, all historical data are expressed in current terms; inflationary effects are taken into account. SOFT DRINKS OFF-TRADE RTD VOLUME 534. 8 billion litres Bottled Water 192 billion litres Sports and Bottled and Energy Drinks Sports Energy Water 15 billion litres Drinks 205. 1 billion 16. 2 billion litres Concentrates litres 43 billion litres RTD Coffee 4. 5 billion litres Disclaimer Much of the information in this briefing is of a statistical nature and, while every attempt has been made to ensure accuracy and reliability, Euromonitor International cannot be held responsible for omissions or errors. Figures in tables and analyses are calculated from unrounded data and may not sum. Analyses found in the briefings may not totally reflect the companies’ opinions, reader discretion is advised. Carbonates 169. 5 billion litres. Fruit/Vegetable Juice 62. 0 billion litres Concentrates 43. 7 billion litres RTD Tea 30. 1 billion litres While Red Bull remains the world leader in energy drinks, it is facing growing competition from other players. TCCC in particular, with Monster in the US and Burn in Brazil, is also posing an increasing threat. These two markets are emerging as energy drinks battlegrounds and the implications are considerable for Red Bull’s ability to remain the number one ranked player.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 2Dietrich Mateschitz and Chaleo Yoovidhya each owned a 49% stake prior to 2012 when Mr Yoovidhya passed away. Mr Yoovidhya’s son Chalerm holds the remaining 2%. While Mr Yoovidhya was alive he acted as a silent partner. Red Bull GmbH Headquarters: Regional involvement: Category involvement: Fuschl am See, Austria Global Carbonates, sports and energy drinks ? Red Bull has created the global market for energy drinks, and the pioneering Red Bull brand has became synonymous with energy drinks for a large number of consumers. Red Bull remains bullish and ambitious in their corporate brand. Despite rising competition, Red Bull continues to comfortably lead the global energy drinks market in both volume and value terms. However, the threat from The CocaCola Co (TCCC) has been mounting. World soft drinks share by off-trade 0. 2% RTD volume (2012): World soft drinks off-trade RTD volume 12. 4% growth (2011-2012):  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 4 STRATEGIC EVALUATION Red Bull continues to see strong net sales growth ? Red Bull operates many other businesses aside from energy drinks The company owns and manages a construction company, football clubs, youth academies and TV broadcasting and recently online clothing (Red Bull label only) sales. ? Additional media products include print magazines about football, motor racing, celebrity gossip and lifestyle. The company has even ventured into the mobile phone service business in Austria, Hungary, Switzerland and South Africa.? As a privately-held company, financial information is limited however the company reported net sales of â‚ ¬4. 9 billion in 2012 and 5. 2 billion cans sold, representing growth of 15. 9% and 12. 8%, respectively.  © Euromonitor International ? Red Bull reported exceptionally strong net sales growth in South Africa (+52%), Japan (+51%), Saudi Arabia (+38%), France (+21%), the US (+17%) and Germany (+14%). Red Bull cited efficient cost management and ongoing brand investment as underpinning its growing profitability. SOFT DRINKS: RED BULL GMBH PASSPORT 5 STRATEGIC EVALUATION SWOT: Red Bull GmbH STRENGTHS WEAKNESSES Broad geographic presence ? Red Bull has ? Red Bull has a broad established a strong, geographic presence, consistent brand image which should ensure (an independent, edgy positive long-term brand) globally. Red Bull growth even if certain is synonymous with markets reach maturity. energy drinks in many countries. Category leader OPPORTUNITIES Category limitations Controversial ? In overall soft drinks, ? The relatively high Red Bull has a limited caffeine content of Red product portfolio Bull makes the brand compared to the rising highly vulnerable to number of rivals with a regulatory control. plethora of flavour variants and categories. THREATS. Emerging markets New production Competition High marketing costs ? Red Bull is building a new ? Monster represents the ? Market maturity in ? Emerging markets production facility in Brazil biggest threat to Red developed markets will represent newer which is likely to make its Bull as it contains make marketing to its geographies for Red retail price more natural ingredients, core consumers harder Bull’s expansion. competitive than imported which seem more than in the past. Accelerating the product prices. Building a desirable than Red Bull Constant communication marketing and site in Asia should also be for some consumers.with consumers means sponsorships in these high marketing costs. markets is a wise move. considered.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 6 STRATEGIC EVALUATION Key strategic challenges and objectives It is not easy at the top ? Red Bull’s success has attracted considerable interest from soft drinks multinationals, TCCC and PepsiCo. TCCC in particular has been successful at leveraging its distribution network to launch Burn across many markets and to back Monster. Burn is a major threat to Red Bull in Brazil while in the US Monster has overtaken Red Bull in offtrade volume sales terms. Red Bull will need to find ways to hold onto its number one ranking globally in energy drinks and stave off this competition. Will premium work in emerging markets? ? Red Bull has consistently maintained its premium positioning from its slimline metal cans to its price differential versus brands such as Monster. While this strategy has reaped dividends in the mature markets, it remains to be seen if it will sustain growth in the emerging markets. Brazil with its large population of lower-income consumers may pose a challenge giving cheaper brands such as TCCC’s Burn a competitive advantage. Red Bull stands up to health regulators ? While health officials continue to voice concerns over energy drinks and the category remains under threat from stronger regulation, energy drinks has seen relatively little impact in terms of sales. To some extent this has added to the category’s â€Å"edginess† attracting young consumers and generating consumer interest. There is little risk of Red Bull reformulating its product to cater to health concerns and instead the company insists that its products do not pose a health risk. Red Bull breaks with tradition in 2013 ? In 2013, Red Bull, for the first time in 15 years added new products to its energy drinks range. Red Bull underperformed the overall energy drinks market in 2011-2012. While the company’s market share of the energy drinks market in the US increased in 2012, the market’s growth rate overall began to wane. Red Bull remains heavily dependent on the US for its global growth. Weakness here is reflected in the company’s weakening global performance in volume terms. The company however continues to enjoy the position of number one ranked player in energy drinks globally with a 21. 4% market share. ? In terms of absolute volume growth however, the US remained Red Bull’s key growth engine in 2011-2012 reflecting growth of 96% over 2007-2012. Brazil came second in terms of absolute volume growth expanding by 608% over the review period or 48% CAGR. This market was a particular focus for Red Bull with the company sponsoring various sporting events in order to raise the brand’s profile.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 9 COMPETITIVE POSITIONING Red Bull faces mounting pressure ? In value terms, the company’s performance was stronger in recent years although even in value terms the company’s performance fell below that of the energy drinks market overall. The energy drinks market has attracted a number of other players including Monster Beverage Co, and The Coca-Cola Co (TCCC) which marketed it own brands in the category including Burn as well as engaging in a distribution alliance with Monster Beverage Co. PepsiCo had a modest presence in energy drinks with its brand Sting; however like TCCC it maintained its own alliance, with Rockstar Inc. ? Red Bull’s sister brand non-carbonated Red Bull remains owned by TC Pharmaceutical which led the energy drinks category in China and was present in Thailand where it ranked second. Top 10 players in soft drinks by off-trade value share Soft Drinks: Global Top 10 Companies by Off-Trade Value, Rank 2007-2012 and 2012 Share 2007 2008 2009 2010 2012 2011 Company Coca-Cola Co, The PepsiCo Inc Nestle SA Suntory Holdings Ltd Dr Pepper Snapple Group Inc Danone, Groupe Red Bull GmbH Asahi Group Holdings Ltd % company share 2012 26. 2 11. 3 2. 8 2. 7 2. 0 1. 9 1. 6 1. 5 ? Danone’s volume share is significantly higher than its value share, due to its large volume sales of low-priced bottled water in emerging markets, notably Aqua (Asia Pacific) and Bonafont (Latin America). Meanwhile, Mondelez does not rank among the top 10 in value terms due to its reliance on the low-priced concentrates category in RTD volume terms. ? Red Bull GmbH however with its relatively premium but small serving size Red Bull brand ranks seventh in 2012. The company’s narrow focus in soft drinks, being almost exclusively based on energy drinks, continues to keep the company out of the top five in soft drinks. ? TCCC and PepsiCo capture a stronger share in value than in volume terms chiefly due to their products, particularly carbonates, being priced higher than local brands and   ? North America will continue to lead energy drinks in absolute volume growth terms over the forecast period. However, its CAGR of 8. 1% over 2012-2017 represents a moderation from the 11. 4% CAGR seen over 2007-2012. The Monster brand has led the market in the US over the review period in terms of absolute volume growth. Rockstar, due in large part to its alliance with PepsiCo, has also seen strong growth in this market. ? Red Bull entered China in 2011, however Asia Pacific remains the company’s weakest region in terms of market share. However, this region will be exceeded only by North America in terms of absolute off-trade volume growth over 2012-2017 which may raise some concerns for Red Bull. After a period of strong market share gains in this region between 2007-2010 its performance began to moderate. TC Pharmaceutical with its non-carbonated version of Red Bull is the regional leader. Despite the close relationship between Red Bull GmbH and TC Pharmaceutical with the latter having been founded by the late Chaleo Yoovidhya, the companies remain separate entities.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 14 MARKET ASSESSMENT The Americas to lead growth in energy drinks ? In value terms, both Latin America and Asia Pacific gained in importance for Red Bull over the review period. Latin American sales represented 12% of global value sales in 2012 while Asia Pacific made up 8%. In terms of growth prospects, the strongest growth will take place in North America where the market for energy drinks will expand by US$4. 1 billion over 2012-2017. In CAGR terms however, the strongest performance will take place in Latin America which will see a 20% CAGR. ? Red Bull is ranked number one in both markets. In Latin America, its market share remains a healthy 49. 7%, however this represents a decline over 2007-2012 as the company faced strong competition from TCCC whose share has risen from 2. 5% in 2007 to 14. 9% in 2012. ? Growth in both Eastern and Western Europe will be a comparatively modest at 5% and 5. 1% CAGRs, respectively. However, these exceed the CAGRs for soft drinks overall in these regions, which will be only 2. 7% and 0. 5%, respectively. CATEGORY AND GEOGRAPHIC OPPORTUNITIES Leading players in energy drinks by off-trade volume and value  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 17 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Red Bull shows some weakness in volume sales ? The rankings of the leading players in energy drinks vary significantly by volume and value. Red Bull commands a stronger market share in value than in volume terms reflecting its relatively high price points and reliance on the mature markets, particularly the US, for its sales. The company however maintained is leading position by both measures in 2012 although in both cases it has seen its market share plateau over 2007-2012. ? The Lucozade brand has faced strong competition in its domestic UK market from Red Bull. In 2012, GSK announced a strategic review of the Lucozade and Ribena brands, which may lead to possible divestment. ? Red Bull has been constrained to some extent in volume terms by its highly concentrated production infrastructure. Up to 2012, the company produced exclusively in Austria leading to high shipping and ? The major winner over the review period was production costs, which opened up the emerging Monster Beverage Co, which until 2012 was known markets in particular to less expensive energy drinks as Hansen Natural Corp. Underpinned by its brands. In 2012, the company announced plans to distribution agreement with TCCC the brand has build its first factory abroad in Brazil which may help made rapid gains in both value and volume terms. improve its competitiveness. The brand’s success has been driven by its North ? Rockstar’s distribution agreement with PepsiCo did American performance where it generated 90% of not bring in the same share gains as the Monster its volume sales in 2012. and TCCC alliance. Rockstar made few share gains ? In contrast GlaxoSmithKline (GSK) and its Lucozade brand have been losing market share. In volume terms, GSK has lost 2. 4 percentage points in market share over 2007-2012. globally, with sales mainly coming from developed Western markets where Red Bull continues to lead. PepsiCo may have found it hard to drive Rockstar sales in these mature markets in the face of TCCC’s penetration and Red Bull’s dominance.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 18 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Most dynamic energy drinks markets over forecast period  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 19 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Red Bull tries to counter weakness in key markets with new launch ? While the US will lead growth in energy drinks in both volume and value terms over 2012-2017 there are clear differences among the top 10 rankings by both measures. ? China will push ahead of Brazil in volume growth terms. The market for energy drinks in China is more mature than in Brazil. Unit price growth in Brazil will as a consequence be higher than that in China allowing it to take second position in terms of value sales growth. In China, Red Bull’s sister company TC Pharmaceutical with its Red Bull is the overwhelming category leader with a market share of 81. 2% in off-trade volume terms in 2012. ? Markets entering the top 10 in volume terms include the Philippines and Vietnam both relatively price-sensitive markets. Per capita consumption however in both markets is higher than the global average. Energy drinks in many Asian markets have a long history of being consumed by truck drivers and labourers as a temporary energy boost. These products were in fact the original inspiration for Red Bull; a Westernised version of the potent drinks sold through by Thai pharmacists.  © Euromonitor International ? The UK ranks among the top five most dynamic markets in both volume and value terms. While Lucozade remains the leader here, its fortunes have waned. Red Bull was responsible for much of Lucozade’s market share loss in the early part of the review period. However, later in the review period, smaller brands are increasing fragmentation. The UK is becoming increasingly fragmented as newer and smaller players have entered the market. ? In 2013, Red Bull launched three new flavour variants in the US market. This marks the first major launch for the brand in the energy drinks category over the review period. The new range called Edition includes cranberry-, blueberry- and lime- flavoured variants packaged in red, blue and silver cans, respectively. The move may help to invigorate consumer interest in key markets such as the UK and the US where the range of energy drinks options has increased considerably. It is recommended that the range be rolled out to other markets where market share has weakened. SOFT DRINKS: RED BULL GMBH PASSPORT 20 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Top US brands in energy drinks  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 21 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Red Bull and Monster look to high-adrenaline sports sponsorship Monster pulls ahead in volume sales ? The Monster brand pulled ahead of Red Bull in the US energy drinks market in 2009 in volume sales terms but remains second to Red Bull in value terms. Monster has achieved wider presence in supermarket and forecourt retailers. TCCC has leveraged its strong distribution network through both channels thus giving Monster an edge in terms of volume sales. ? The Monster brand has also been supported by sponsorship of high-adrenaline sports such as MotoGP, NASCAR and Freestyle Motocross which is a direct challenge to Red Bull, which also relies on sponsorship of these sorts of events to maintain consumer interest. Another reason behind the disparity has been the fact that Red Bull sells primarily in smaller 8. 3oz cans, whereas Monster is sold in larger 16oz cans at a relatively cheaper price. Red Bull has since begun to offer its product in a wider variety of sizes and in 2012 trumped Monster with Red Bull Stratos, sponsoring Felix Baumgartner’s free-fall from over 128,000 feet. Threat from consumer health ? Both Monster and Red Bull have also been challenged by the 5-Hour Energy brand from Living Essentials, included in Euromonitor International’s Consumer Health database as a tonic and bottled nutritive drink. This product has been heavily marketed on US television and offers a small pack size (57ml) and the benefit of being sugar-free. While Monster is targeted primarily at younger male consumers, 5-Hour Energy is positioning itself as a pick-me-up for office workers and working mothers. ? The addition of new flavours in 2013 will help to reignite consumer interest. Red Bull’s success in the US has been due in part to its success in the on-trade which has helped to introduce the brand into the off-trade. Educating consumers about how the new flavours can be mixed with alcoholic drinks in the on-trade should form part the marketing campaign to launch the brand.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 22. CATEGORY AND GEOGRAPHIC OPPORTUNITIES Leading players in Brazilian energy drinks  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 23 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Red Bull vs Burn in Brazil TCCC pushes Burn ? Strong growth in the Brazilian energy drinks market has attracted a wider number of players, many of whom have focused on the emergent C socioeconomic class, launching energy drinks at lower prices in 1-litre PET bottles. Examples include BadBoy Power Drink from Horizonte and Orbit from Bebidas Chiamulera. These moves have helped to fuel growth overall in the category.? TCCC has made significant gains in the market with its Burn brand investing significant resources in marketing. Like Red Bull, TCCC has targeted high-adrenaline sporting activities, announcing in 2012 its sponsorship of Kimi Raikkonen’s Lotus F1 team. The brand competes directly with Red Bull, packaged similarly in a slimline metal can. Its price points however are typically lower than those of Red Bull giving it a stronger presence among lower-income groups. ? In 2012, Red Bull announced plans to begin producing its energy drinks locally. Localisation of production will help Red Bull ? Localising production in such a key market is a wise move for Red Bull. It also gives the company stronger capacity more widely in Latin America where the markets for energy drinks in Colombia and Mexico are also set to see strong growth. While Red Bull’s number one position remains safe for the time being, reducing the price premium with TCCC is recommended. This will be supported by significantly reducing costs associated with importing the product from Austria. ? The entry of Anheuser-Busch InBev NV was a key development in the market in 2011. By 2012, the Fusion brand had managed to capture 0. 2% of sales in off-trade volume terms which, while modest compared to the Red Bull brand at 19. 8%, indicates strong potential for further growth. Marketing initiatives centred around the popular Big Brother Brazil TV programme in 2012 helped to increase awareness of the brand among young people.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 24 CATEGORY AND GEOGRAPHIC OPPORTUNITIES Worlds apart: A tale of two Red Bulls ? The relative weakness of Red Bull from Red Bull GmbH in Asia Pacific is due in part to the strength of sister brand Red Bull from TC Pharmaceutical. A more cohesive international strategy should be developed by both companies. ? The strongest prospects for the two players is in China, however opportunities are also being missed in markets such as the Philippines, Thailand and Indonesia. TC Pharmaceutical sales here in energy drinks have been virtually flat over the review period, as newer, more dynamic brands such as Cobra from Asia Brewery and Sting from PepsiCo in the Philippines have invested heavily in marketing and advertising. ? A decisive entry for Red Bull GmbH in key Asian markets will be complicated by the presence of TC Pharmaceutical’s Red Bull. However, both companies could benefit from working more closely together including on the production side to reduce costs and widen their distribution network. The sudden death of TC Pharmaceutical founder Chaleo Yoovidhya in 2012 may present a challenge however in ongoing collaboration.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 25 STRATEGIC EVALUATION COMPETITIVE POSITIONING MARKET ASSESSMENT CATEGORY AND GEOGRAPHIC OPPORTUNITIES BRAND STRATEGY OPERATIONS RECOMMENDATIONS BRAND STRATEGY Red Bull’s premium focus will result in pressure on market share ? Red Bull’s sales in 2012 remained dominated by the US market. In most of its major markets the company has managed to retain its number one position in volume terms despite strong competition from newer entrants. The US is an exception where Monster owing to the strength of its alliance with TCCC combined with an aggressive marketing campaign has managed to topple Red Bull from first place. ? In value terms however, the company’s premium positioning has meant its ranking has remained more secure. As the dynamics of forecast demand shift to emerging markets, where consumers remain more price sensitive, this premium focus will result in growing pressure on Red Bull’s market share. TCCC and PepsiCo have emerged as the company’s strongest competition whether indirectly through distribution agreements such as TCCC/ Monster and PepsiCo/Rockstar and through their own directly owned brands such as Burn and Sting, respectively  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 27 BRAND STRATEGY High octane sports drive home Red Bull message ? Event and sports sponsorship have been key elements for Red Bull’s marketing strategy for many years. Red Bull’s eponymous brand has achieved remarkable global success and 30-40% of its sales are re-invested back in marketing and promotional activity. Red Bull’s strategy has historically been a 3-pronged approach incorporating buzz marketing, sponsorship and TV advertising. Buzz marketing, including handing out free samples at campuses and events where under 30s gather, is often used as a way of initially raising consumer awareness when entering new markets. ? In 2012, the company took its marketing literally to an entirely new level with the Stratos campaign which featured Felix Baumgartner in a record- breaking 128,000 feet jump from the earth’s stratosphere, making him the first man to break the speed of sound while in freefall. The event was streamed live on line with viewers able to log in to post comments via Twitter and Facebook. Motorsports is another key focus for the company with its own very successful F1 racing team.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 28 BRAND STRATEGY Red Bull tries to stay true to its roots ? In a bid to stave off competition from rival brands, Red Bull launched the Red Bull Edition range in 2013 in select city markets in the US. The launch will likely be followed by a nationwide roll-out later in the year. Despite pressure from other energy drinks brands many of which have launched additional flavours Red Bull has stayed loyal to its original formulation and packaging. ? The launch of cranberry, blueberry and lime Red Bull variants is a major direction change for the brand, being its first major launch over the review period. In order to differentiate between Red Bull Edition and the original Red Bull the new cans received a facelift with the addition of new colours and a new bull design. ? Red Bull has not as aggressively as other brands launched into new packaging formats, remaining almost exclusively with slimline metal cans. It has however in some mature markets such as the UK launched into 1-litre PET bottles. This reluctance is in part due to the company’s strategy of retaining its premium positioning.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 29 STRATEGIC EVALUATION COMPETITIVE POSITIONING MARKET ASSESSMENT CATEGORY AND GEOGRAPHIC OPPORTUNITIES BRAND STRATEGY OPERATIONS RECOMMENDATIONS OPERATIONS Expanded corporate operations Red Bull GmbH Red Bull Soft Drinks Other Businesses Red Bull Energy Drinks Motor Racing, Media, MVNO, Fashion Online Retailing Red Bull Simply Cola, Carpe Diem.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 31 OPERATIONS Red Bull looks to diversification ? Red Bull is diversifying into other businesses, rather than limiting itself to energy drinks. In recent years, it has been branching out and became a media company in its own right. The participation in sports sponsorships and events connects the company with a global brand that has passion and excitement associated with it. The company is also present in RTD tea and bottled water with the Carpe Diem brand which it launched to target the health and wellness trend in soft drinks. Carpe Diem Kombucha is a premium RTD tea sold in Western Europe. The brand is also in bottled water in Switzerland and Austria using plant extracts and slight carbonation to offer a healthy alternative to carbonates. ? The company owns two Formula One teams (Red Bull Racing, Scuderia Toro Rosso), a NASCAR racing team as well as several football teams in Brazil, the US and Germany. ? In South Africa, the company is partnering with Cell C to offer voice and broadband services as a mobile virtual network operator (MVNO), ie a company that provides a mobile phone service but does not have its own licensed frequency. Red Bull Mobile will be the second MVNO in the country, after Virgin Mobile. ? It also sponsors many events from cliff diving to air races and subscribing to Red Bull Mobile is a way for people who like the brand to access further benefits when they attend these events. These kinds of partnerships between operators and consumer brands are common in Europe. In Germany, for example, one operator, E-Plus, has 19 such partnerships. It is a way for these brands to get closer to their target group. ? The Group also includes Austrian TV station ServusTV, lifestyle and fashion magazines and a construction company called Bull Bau. ? Red Bull had 8,966 employees in 165 countries as of 2012. The company, which is not listed, traditionally finances its investments from its cash flow.  © Euromonitor International SOFT DRINKS: RED BULL GMBH PASSPORT 32 OPERATIONS Red Bull expands production outside Austria for first time ? Red Bull received approval from the B\razilian government to build its first production facility in the country in early 2010. The companys initial investment in the pr.

DNA Based Biosensor in Diagnosis: A Review

DNA Based Biosensor in Diagnosis: A Review The advent of fast and easy DNA testing has given the space for the Science to develop small and easy-to-handle equipments called Biosensors. DNA based biosensors have been proven very useful and are accorded with much importance in detecting the target genes responsible for diseases. This article enlists different types of biosensors, their basic principle of operating system, the preparation of DNA microarrays, lab-on-a-chip and their role in diseases diagnosis. DNA biosensors provide swift, sensitive, selective, simple and economical detection of DNA hybridization. New strategies for DNA biosensor are enumerated and are used meticulously in recent trends and for future directions. Carbon nanotubes (CNTs) amplify the electrochemical signal when used with DNA hybridization. Electrochemical, piezoelectric, SPR, optical DNA biosensors are used to detect various viruses like hepatitis virus, HCMV, HIV, orthopox virus etc. and also for the diagnosis of various diseases like cancer, tube rculosis, COPD, genetic diseases (sickle cell anemia i.e. due to single point gene mutation), cystic fibrosis, diabetes etc. The methodologies of detecting such diseases using different types of DNA based biosensors and gene chips are described in this article. PCR free DNA chips, cell- omic sensors and nanosensor are emerging tools in the field of diagnosis. Recent advances in developing such devices provide myriads of new opportunities for DNA diagnostics. Introduction A rapidly developing area of biotechnology arousing intense scientist interest is that of biosensor. Biosensor has become popular in the field of food analysis [1], bioterrorism [3], environmental [2-3] and in the area of human health monitoring and diagnostics [4-6]. Recent advances are being mad in all areas of biosensors technology. Presently, most fascinating and prospective sensors are immunosensors based on affinity reactions between antibody and antigens and DNA biosensors based on the hybridization between DNA probes and their complementary DNA strands. In general, biosensor is an analytical device which employs biological recognition properties for a selective analysis. Such sensors combine a biological element with a physiochemical transducer for the electronic signal output which is proportional to the concentration of analytes [7]. A basic biosensor assembly includes a biological element, transducer and detector. The sensing material may be antibodies, enzymes, whole cell or nucleic acids that form a recognition layer which is integrated with the transducer via immobilization by cross linking, adsorption or covalent binding. Transducers may be amperometric (measuring the current at constant potential) [8], potentiometric (measuring the potential at constant current) [9], piezoelectric (measuring the changes in mass), thermal (measuring the changes in temperature) [10] or optical (detects changes in transmission of light) [11]. The interaction between the analyte and the biological material, used in biosensors may be of two types: a) Bioaffinity sensors: depend on the selective and specific attachment of the target molecule to the surface-attached ligand partner (e.g. antibodies, nucleic acids). b) Biocatalytic sensors: an immobilized enzyme is used as a tool to recognize the target substrate (sensor strips with immobilized glucose oxidase used for personal monitoring of diabetes). A number of steps, much labor, time and costly instruments are required in usual analytical technique whereas biosensors are economical, fast and simple and can be used in small laboratories and hospitals of remote areas which are devoid of sophisticated instruments facilities. DNA Biosensors Nucleic acid recognition process is the basis of DNA Biosensors. These are being developed with a rapid pace with an ambition for inexpensive testing for genetic and infectious disease and for detecting DNA damage and interactions. The study of gene polymorphisms and the analysis of gene sequences play a fundamental role in rapid detection of genetic mutations, opens up new opportunities for reliable diagnosis even before any symptoms of a disease appear. Thus recent advances in developing such devices offer the opportunities for DNA diagnostics. DNA biosensors are made by immobilizing single stranded (ss) DNA probes on different transducers for measuring the hybridization between the DNA probes and their complementary DNA strands [12-13]. The current methods to identify specific DNA sequence in Biological samples depends on the isolation of double stranded (ds) DNA and further polymerase chain reaction (PCR) to amplify the target sequence of DNA. The PCR product is then subjected to electrophoresis or adsorbed onto a suitable membrane and exposed to a solution containing DNA probe. Surface Chemistry and Biochemistry The immobilization of DNA probe onto the transducer plays an important role in the performance of the DNA Biosensor. It should be in well-defined probe orientation and should be readily accessible to the target. The mode of immobilization is the determining factor for the type of environment of probes that are immobilized at the solid surface. On the basis of nature of physical transducer, various schemes can be opted for the DNA probes attachment to the surface such as thiolated DNA utilisation for self binding onto gold transducers, the formation of a complex by the use of biotylated DNA with a surface-confined strepavidin or avidin, covalent binding to the gold surface through functional alkanethiol-based monolayer and coupling covalently (carbodiimide) to the functional groups on carbon electrodes or adsorption onto carbon surfaces. Introduction of peptide nucleic acid (PNA) has paved way for many exciting and new opportunities to DNA biosensors. Peptide Nucleic Acid is a DNA mimic, the only difference is that the sugar-phosphate bone is replaced by a pseudo-peptide one. Like use of surface-confined PNA recognition layers provides remarkable sequence specificity on DNA biosensors and offers other advantages. DNA dendrimers may also be utilized for imparting extreme sensitivity onto DNA Biosensors. By shape, these are tree-like superstructures which possess numerous ss arms that are able to hybridize to their complementary DNA sequence. The immobilization of these dendritic nucleic acids onto physical transducer gives an amplified response [14]. Recent advances in the field of biomolecular techniques may be used to design new generation miniaturized biosensor. Types of DNA based Biosensors 1. Optical Type Fiber optics Biological Element Laser Interferometry Transducer DNA Advantages Optical fiber Highly sensitive Disadvantages Expensive equipment and not portable turbidity interference 2. Electrochemical Type Potentiometric Biological Element Conductometric Transducer Amperometric DNA Advantages Carbon paste electrodes Cheap, Fast Limitations Interference of highly buffered solution 3. Piezoelectric Type DNA Biological Element Quartz Crystals Advantages highly sensitive, Fast 4. DNA chips DNA Quantitative Optical DNA based Biosensor Optical methods are the most commonly used for the detection of analytes. DNA optical biosensors are based on a fiber optic which transduces the emission signal to a fluorescent label and that can carry light from one region to another through a series of internal inflections. The methodology of fiber-optic DNA bio-sensors involves placing of a single stranded DNA probe at the ending-site of fiber and assessing the fluorescent changes resulting from the combination of a fluorescent indicator with the double stranded DNA hybrid [15 16]. The first DNA optical bio-sensors were developed by Krull and Co workers using fluorescent indicator ethidium bromide. A fiber-optic DNA sensor array was developed by Watts group for the detection of multiple DNA sequences at one time [17]. The hybridization of fluorescent labeled complementary oligonucleotides was assessed by observing the increase in fluorescence. A real label free optical detection of DNA hybridization can be offered by a different type of optical transduction based on evanescent wave devices. The different types of optical biosensors include: 1.1 Surface Plasmon Resonance (SPR) It is a quantum optical electrical phenomenon based on the interaction of light with metal surface. Only at specific resonance wavelength of light, the energy carried by photons of light is transferred to packets of electrons (photons) on a metal surface [17]. These biosensors depend on change in surface optical properties (change in resonance angle because of alteration in interfacial refractive index) which results from the surface binding reaction. Thus, these devices integrate the simplicity of SPR with the sensitivity and specificity of wave guiding devices. The SPR signal that is expressed in resonance units is therefore a measure of mass concentration at the senor chip surface [18-20]. 1.2 Molecular Beacons (MBs) MBs are oligonucleotides possessing a stem and loop structure that are labeled with a quencher at one end and a fluorophore on the other end of the stem that converts into fluorescent upon hybridization. MB probes possess high sensitivity and specificity and direct monitoring capability. A biotinylated molecular beacon probe was developed to prepare a DNA sensor using a bridge structure. MB was biotinylated at quencher site of the stem and linked on a glass through streptavidin that act as a bridge between MB and glass matrix. The fluorescence change was measured by confirmation change of MB in the presence of complementary target DNA [21-23]. Quantum Dot It is an ultra sensitive nanosensor based on fluorescence resonance energy transfer (FREET) that can detect very low concentration of DNA. In these neon sensors, quantum dots (QDs) are linked to specific DNA probes to capture target DNA. The target DNA strand binds to a fluorescent dye (Fluorophore) labeled reporter strand and thus forming FREET donor acceptor assembly. Quantum dot also functions as target concentrator as well as FREET energy donor [24]. DNA nanosensor contains two target specific DNA probes i.e. reporter and capture probe. The reporter probe is labeled with fluorophore whereas capture probe is labeled with biotin that binds with streptavidin conjugated with QD [25]. The fluorophore acceptor and QD donor in close proximity produce fluorescence from acceptor by means of FREET on illumination of the donor. The presence of target DNA is indicated by the detection of acceptor emission. The un-hybridized probe does not give fluorescence. The CdSe Zns core shell nanocrys tal can be used as donor and Cy5 (fluorophore) as acceptor for developing QD based DNA nanosensors [25]. For this type of optical bio sensors fluorescent dyes used as standard labels are very expensive and can rapidly photo bleach. An alternate used is chemiluncinscence format, which overcomes the use of fluorescent dyes. A Fiber-optic DNA biosensor array A new method of preparing the fiber-optic DNA biosensor and its array for the simultaneous detection of multiple genes is described. The optical fibers were made into fiber-optic DNA biosensors by adsorbing and immobilizing the oligonucleotide probe on its end but were first treated with poly-l-lysine. The fiber-optic DNA biosensor array was well prepared by assembling the fiber-optic DNA biosensors in a bundle in which each fiber carried a different DNA probe. Hybridization of fluorescent- labeled cDNA of Rb1 gene, N-ras gene and Rb1 p53 gene to the DNA array was monitored CCD camera. A good result was achieved [61]. 2. Electrochemical DNA Bio sensors These are very useful devices for sequence specific biosensing of DNA. The inherent miniaturization of such devices and advance micro fabrication technology make them excellent tool to diagnose DNA. DNA hybridization is detected electrochemically by monitoring the current response at fixed potential. Detection of hybridization is also commonly done through the increased current of a redox indicator or from other changes induced by hybridization in electrochemical parameters such as capacitance or conductivity [26-28]. The discovery of carbon nano tubes (CNTs) plays an important role in development of electrochemical DNA sensors. Various CNT based electrochemical are developed because the combination of unique electrical, thermal, chemical, mechanical and 3-D spatial properties of CNTs with DNA hybridization offers the possibility of creating DNA bio sensors with specificity, simplicity, high sensitivity and multiplexing. Two major groups in which CNTs divided are single walled CNTs (SWCNTs) that are comprised of a single graphite sheet rolled with a tube and multi walled CNTs (MWCNTs) that are concentric closed graphite tubes [29]. CNT enables immobilization of DNA molecules and also used as powerful amplifier to amplify signal transduction of hybridization [30]. Two types are generally used to immobilize the CNT on electrodes aligned and non-aligned. Two approaches are generally used for the immobilization of bio molecules onto CNTs that are non covalent attachment (physical absorption) and covalent binding (some cross linker agents (1-ethyl 3-3 dimethylaminopropyl) carbodilimide hydrochloride (EDC)/N-hydroxysuccinimide (NHS)] or affinity binding (avidin biotin interaction). CNT also act as novel indicator of hybridization. The application of arrayed CNT into DNA chip requires small amount of sample and development of CNT base biosensor has an important role in DNA based diagnostics in hospitals or at home [30]. Various methods are used for immobilizations step i.e. for attaching the DNA probe onto the solid surface that are (a) the use of thiolated DNA probe for self assembled monolayers (SEM) onto gold transducers by covalently bonding to the gold surface through functional alkanethiol based monolayers.(b) Attachment of biotinylated DNA probe through biotin avidin interaction on electrode surface for e.g. avidin modified polyaniline electro chemically deposited onto a Pt disc electrode for direct detection of E. Coli by immobility a 5 biotin labeled probe using a differential pulse Voltametric technique in the presence of methylene blue as an DNA hybridization indicator [31,32]. The electrochemical DNA biosensors may be labeled free and labeled based. Label Free In this direct detection technique the target molecule does not need to be labeled [27]. The elimination of labeling steps simplifies the readout the speed and ease of nucleic acid assays. Thus recently increased attention has been accorded to new label free electrochemical detection schemes. There is a possibility in exploiting the changes in DNAs intrinsic electroactivity (Guanine oxidation peak of hybridization). To deal with the drawbacks of the probe sequences i.e. absence of G, guanines were substituted by inosine residues (pairing with C) and detection of hybridization was done through the target DNA guanine signal. The change in the guanine oxidation and intrinsic DNA redox signals detects the chemical and physical damage [33]. Label Based In label based electrochemical biosensor specific organic dyes, enzymes or metal complexes are used for hybridization detection. Redox active molecules such as methylene blue, dacinomycin that is inserted between the dsDNA and gives signal which is used for detecting hybridization [26] (e.g. of two commercialized DNA chips based on redox active molecules are eSensor TM produced by Motorola life sciences [34], Inc. and Genlyser TM by Toshiba)[35]. Piezoelectric DNA Biosensor These are the mass sensitive devices rely on quartz crystal that oscillate at a defined frequency when oscillation voltage is applied. Increased attention has been given to piezoelectric method due to their simplicity, cost, sensitivity and real time label free detection. The quartz crystal microbalance is an extremely sensitive piezoelectric device that monitors the hybridization events. These biosensors DNA probe is immobilized on the surface of oscillation crystal. The increased mass due to hybridization reaction results in change in oscillating frequency [36-37]. A Piezoelectric sensor for determination of genetically modified soyabean roundup ready [RR soyabean] by immobilizing probe related to 5-enolpyrllvylshikimate 3-phosphate synthase (EPSPS) gene onto gold piezoelectrodes [38]. For detecting a point mutation in a human gene (apolipoprotein-E polymorphism) a combination of DNA piezoelectric biosensor and PCR was developed by immobilizing biotinylated probe on the streptavidin coated gold surface of quartz crystal. The hybridization probes with complementary, non-complementary and mismatched DNA of synthetic as well as amplified PCR samples from human blood DNA was taken out and the device was able to distinguish polymorphism [39]. Colorimetric or Strip type DNA sensor Using these sensors the direct detection of DNA hybridization is possible [40-42]. The dry-reagent strip type biosensor has been developed for visual detection of double stranded DNA within a short time [43]. Oligonucleotides conjugated gold particle is used as probe. The main advantage of these biosensors is not requiring any instruments, multiple incubation and washing steps. Integral part of strip consists of gold particles, with oligo (dT) attached to their surface. Biotinylated PCR products are hybridized with poly (dA) tailed oligo, switched to the top of strip and immersed in the appropriate buffer. With the migration of buffer in upward direction, the nanoparticles that are linked through target DNA through poly (dA/dT) hybridization are rehydrated. Immobilized streptavidin then capture the hybrid in the controlled zone of the strip. The test is 8-10 times more sensitive than ethidium bromide in agarose gel electrophoresis. The detection limit is abysmally low of 2 fmol of amplified DNA products. DNA Biochips Microarrays, DNA arrays, gene chips or biochips are same terminology often being intermixed. DNA microarrays are small, solid supports which themselves are usually microscopic slides, but can also be silicon chips or nylon membranes onto which the sequences from thousands of different genes are immobilized, or attached, at fixed locations. The DNA may be spotted, or synthesized directly onto the support. DNA microarrays detect the change in gene expression levels, genomic gained and losses, mutations in DNA and infectious agents, diagnosis of genetic diseases, drug screening or forensic analysis. Developing the methods for detecting target hybridization, designing probe arrays, data analysis and reconstructing the target sequence are required for successful implementation of DNA chip technology. Such array technology thus forms the basis of integration of molecular biology, surface and analytical chemistry, advanced micro fabrication, robotics, software and automation. In this technique, RNA extracted from two samples are labeled with two different fluorochromes (generally the green cyanine 3 and the red cyanine 5 (Cy3, Cy5)) before being hybridized to a biochip consisting of large numbers of cDNAs/oligonucleotides arranged orderly onto a glass microscopic slide. After hybridization, a scanner records excitation of the two fluorochromes at given wavelengths and the intensity of the fluorescence emission signals that is proportional to transcript levels in the biological samples. The data is analyzed using specific software that enables clustering of genes with similar expression patterns, with the assumption that they share common biological functions [33, 44]. A new ultrasensitive electronic sensor has been developed by Singapore scientists that would speed up effectively DNA testing for disease diagnosis and biological research. The novel electronic sensor array would be faster, accurate and cost-efficient. Excellent sensitivity has been shown by the Nanogap Sensor Array in detecting the trace amounts of DNA. By saving time and cutting expenses, newly developed Nanogap Sensor Array offers a scalable and viable alternative for DNA testing. The presence of DNA is translated into an electrical signal by biosensor for computer analysis. The distinctively and meticulously designed sensor chip has the ability to detect DNA efficiently. The novel vertical nanostructure design and two different surfaces of the sensor allow ultrasensitive detection of DNA [45]. Lab-on-a-chip (LOC) Lab Chip is a device which involves preparation of sample and detection of DNA array. The objective of this technology is to integrate multiple processes, including collection of sample and pretreatment of it with the DNA extraction, hybridization and detection, on single self-contained microchip i.e. on a microfluidic platform. The capability to do all the processes on a single chip merits excellent advantages in terms of cost, speed, efficacy, effectiveness, contamination, sample consumption and automation. Laboratory transportation to the source of sample will be enabled by such miniaturization of analytical instrumentation. The development of these credit-card sized microlaboratories is commonly based on latest micromachining and microfabrication technologies, utilizing processes well known in the manufacture of electronic circuitry [14]. Cell-omic sensors Cell based detection systems can be combined with the microarray probes generating the hybrid arrays of cells within arrays of DNA/protein probs. This allows multiparameters analysis [46]. Applications of DNA Biosensors Biosensors plays a distinguished role in the field of environmental quality, food analysis, study of biomolecules and their interactions, drug development, crime detection, medical diagnosis, quality control, industrial process control, detection system for biological warfare agents, manufacturing of pharmaceuticals and replacement organs. The applications of DNA biosensor can be classified into three broad categories: sequencing, mutation detection and matching detection [47]. Their main use is for diseases diagnosis. Numerous diseases can be diagnosed and variety of infectious agents can be detected using DNA biosensors. 1. Viral diseases By DNA microarrays Either viral detection were being carried by immunological techniques (i.e. use of enzyme-linked immunosorbent assays (ELISAs) for the detection of circulating virus-specific antibodies) or PCR based techniques (i.e. reverse transcriptase (RT) PCR is used to detect the presence of specific viral genes). Both these approaches possess some limitations. Immunological tests need specific antisera and the production of antisera is laborious and time-consuming task whereas PCR is prone to failure in its ability to identify multiple viruses simultaneously [48]. Therefore, recent advances in DNA and protein microarray methodology fulfill the need of a rapid and sensitive detection of viral infections (also identify multiple viruses in parallel). DNA microarrays for viral analysis can be divided into viral chips and host chips. Each not only detects and identifies but also monitor the viral populations. In 1999, the first viral DNA microarray for the temporal profiling of viral (human cytomegalovirus, HCMV) gene expression was described. Viral replication or de novo protein synthesis was blocked by treatment of infected cells with cycloheximide or ganciclovir and then the expression profiles of viral genes was generated using microarray. Using this approach, the HCMV genes were classified to immediate-early, early or late expression classes, on the basis of their expression profile in response to the drug treatments. This can be used as an identifying hybridization signature for the molecular staging of an infection [49]. Orthopoxvirus causes smallpox and has two subtypes variola major and variola minor, of differing pathogenicity. This problem of orthopoxvirus subtype discrimination was solved by producing an array capable of correctly identifying the four of the orthopoxvirus species by laassri etal. [50]. HIV genotyping was done using chip technology [51]. A unique signature that is derived from viral is provided by viral chips. Host chip is used for examining the host response i.e. changes in host gene expression. This provides a molecular signature of infection. Cummings and Relman exposed an idea of host chips [52]. Vant wout etal. examined HIV 1 infection in CD4+ T-cells to detect changes in host gene expression that were specific to HIV infection [53]. Proinflammatory genes and genes involved in endoplasmic reticulum stress pathways, cell cycle, and apoptosis were the host gene signatures identified. Detection of hepatitis B virus Hepatitis B virus (HBV) is one of the causative agents of viral hepatitis which is leading cause of liver cancer. Infection of HBV is a public health menace for worldwide resulting acute and chronic clinical consequences. Acute HBV infection may lead to liver failure or may progress to chronic liver disease. Some chronically infected individuals may subsequently suffer cirrhosis and liver failure or develop hepatocellular carcinoma. Effective antiviral therapy may inhibit or retard the progression to severe liver disease. By DNA optical biosensor Bacterial alkaline phosphatase (phoA) gene and hepatitis B virus (HBV) DNA were used as target DNA. For capturing the target gene onto streptavidin coated magnetic beads, a biotinylated DNA probe was used. A calf intestine alkaline phosphatase labeled DNA probe was used for subsequent enzymatic chemiluminescences detection. The detection cycle was less than 30 min, excluding the DNA hybridization time that was about 100 min. at fematomole or picogramme levels both phoA gene and HBV DNA could be detected. No response signal was obtained when in sample target DNA did not exist [54]. By Piezoelectric DNA biosensor HBV nucleic acid probe was immobilized onto the coated gold surface of quartz crystal using polyethyleneimine adhesion, glutaraldehyde cross-linking (PEI-Glu) method or the physical adsorption method. Better results were obtained with the coated crystal with the PEI Glu method to immobilized HBV nucleic acid probe than physical adsorption method with respect to sensitivity, reproducibility and stability. With the hybridization reaction, the mass is increasing that resulted change in oscillating frequency. The frequency shifts of hybridization have better linear relationship with the amount of HBV DNA, when the amount was in range of 0.02-0.14 microgram/ml [55]. By electrochemical DNA biosensor An electrochemical DNA biosensor that is a glassy carbon electrode (GCE) modified with label free21mer single-stranded (ss) oligonucleotides (related to hepatitis B virus sequence) via covalent immobilization. [Cu(dmp)(H2O)Cl2] (dmp = 2,9-dimethyl-1,10-phenanthroline) is used as an electrochemical indicator. The method is simple, economical and allows the accumulation of copper complex within the DNA layer. Cyclic voltammetry and differential pulse voltammetry were used for electrochemical detection. The detection of hybridization is accomplished by using [Cu(dmp)(H2O)Cl2], where electroactivity and strong association with the immobilized dsDNA segment lead to significantly enhanced voltammetric signal. The differential pulse voltammograms for the cathodic signals of [Cu(dmp)(H2O)Cl2] at a bare GCE, and at ss- and dsDNA-modified GCEs are also recorded. The peak currents of [Cu(dmp)(H2O)Cl2] increased in the order of bare GCE, ssDNA/GCE, and dsDNA/GCE. After hybridization process, a greater peak current was observed from dsDNA/GCE than at ssDNA/GCE, because that more [Cu(dmp)(H2O)Cl2] molecules are concentrated or bound to dsDNA helix than to ssDNA. Thus, [Cu(dmp)(H2O)Cl2] can be used as an electroactive indicator for recognition of the surface hybridization process. The sensitivity of the electrochemical hybridization assay was investigated by varying the target oligonucleotides concentration. The different current value obtained in the DPV response of [Cu(dmp)(H2O)Cl2] after hybridization of probe with target is recorded with three repetitive measurements. The current response at about 0.485V increased in proportion to the amount of the target sequence used [56]. Detection of hepatitis C 3a virus An electrochemical DNA biosensor i.e. a gold electrode modified with a monolayer of a peptide nucleic acid probe and 6-mercapto-1-hexanol was used that depends on covalent binding of the14-mer PNA probe (related to the HCV genotype 3a (pHCV3a) core/E1 region) onto the electrode. This self-assembled PNA could selectively hybridize with a complementary sequence in solution to give dsPNA-DNA on the surface, and this increases the peak current of methylene blue (MB) which is used for detecting target DNA sequence. Diagnostic performance of the biosensor is described and the detection limit was found to be 5.7  ÃƒÆ'-  10à ¢Ã‹â€ Ã¢â‚¬â„¢11  M with a relative standard deviation of 1.4% in phosphate buffer solution, pH 7.0. This sensor exhibits high reproducibility and could be used to detect the target DNA for seven times after the regeneration process [57]. Cystic fibrosis Mikkelsens team, pioneered the utilization of redox indicators, demonstrated utility of electrochemical DNA biosensor for detecting the cystic fibrosis F508 deletion sequence which is associated with 70% of cystic fibrosis patients. For the 4000-base DNA fragment, 1.8 fmol was the detection limit in relation to a Co(bpy)33+ indicator. High selectivity for the disease sequence (not for normal DNA) was accomplished by doing the hybridization at high (43 °C) temperature [14]. 3. Diabetes Diabetes is a worldwide public health problem. The diagnosis and management of diabetes requires a tight monitoring of blood glucose levels. Thus millions of diabetics test their blood glucose levels daily by making glucose the most commonly tested analyte. The challenge is to provide such reliable and tight glycemic control. Electrochemical biosensors for glucose thus play a leading role. Amperometric enzyme electrodes, based on glucose oxidase (GOx) bound to electrode transducers, have thus been found the subject of substantial research [58]. Glucose sensors are commonly used to measure the blood glucose level of diabetes patients. Using the latest DNA chip technology, many scientists at Diabetes Center have discovered the implication of new gene in the cause of type 2 diabetes. They created an abnormality in one of these genes known as ARNT (aryl hydrocarbon receptor nuclear translocator gene which is a member of a family of transcription factors) in mice and the mice developed changes in insulin secretion which were same as in patients with type 2 diabetes. The ARNT is required for the development of normal embryo. It is also related to responses to hypoxic stress condition and certain environmental toxins, such as dioxin and thus for integrating genetic and environmental insults it is present at specific potential sites. The expression of many other genes in the cell is regulated by transcription factors like ARNT and thus they are the master regulators of cellular functions. The first use of DNA chips has been represented by this study,

William Shakespeare :: essays research papers

William Shakespeare, undoubtedly one of the best playwrights of all time, was born in April of 1564 in Stratford-upon-Avon in Warwickshire. His parents were John Shakespeare, a whittawer, and Mary Arden Shakespeare. He was the third of eight children born to John and Mary and was their first son. They lived on Henley Street in Stratford. His baptism was on April 26 of that year at Stratford's Holy Trinity Church. His date of birth is assumed to be April 23. He went to Stratford Grammar School from the age of seven to fourteen. At the age of eighteen, on November 28, 1582, William was married by the bishop of Worcester, to Anne Hathaway of Shottery. Anne was seven or eight years older than him and was three months pregnant at the time of their marriage. Their first child together, Susanna, was born on May 26, 1583. Two years after that, Hamnet and Judith were born together. Shakespeare and his family most likely lived in the house on Henley Street. In 1592, Shakespeare's theater career took off. Robert Greene referred to him as an "upstart crow", and from that point on, everyone started paying attention to him. Other famous and notable literary critics also commented on Shakespeare. Shakespeare belonged to a number of acting companies in the beginning: The Queen's Men, Pembroke's Men, and Lord Strange's Men. But in January of 1593, a plague broke out and theaters all across London were closed. In December of 1594, Shakespeare has been noted to have started performing again. His new acting company was known as Lord Chamberlain's Men. The decade or so after that, was huge for Shakespeare. This was the time of The Great Globe theater where many of his best plays were performed. During this time, Shakespeare produced a steady line of hit plays, historical plays, comedies, and tragedies. In 1599 he finally became a part owner of the Globe. While all of this went on in London, Shakespeare's family resided back in Stratford, about 100 miles northwest of London. It is suggested that he often made trips to see his family and deliver them money.

Scientology Essay example -- essays research papers

Scientology is a fairly new religion. Founded in the twentieth-century by a man by the name of L. Ron Hubbard. He began his studies long ago and wrote a book in 1950 called Dianetics: The Modern Science of Mental Health. He claimed that this book was one of the first tools used to solve the problems of the mind. This book focused on irrational mind, war, crime, and insanity. Mr. Hubbard designed this book so that anyone can use it to improve oneself. He did not stop at Dianetics, however, he felt that there were still some issues that had to be addressed. He was now going to try to figure out the nature of the human being, what makes up our â€Å"being.† Mr. Hubbard combined religion, philosophy, and his theory of dianetics and came up with Scientology. Mr. Hubbard had a unique life. He was born in Tilden, Nebraska, the son of a naval commander Harry Ross Hubbard and Ledora May Hubbard. L. Ron Hubbard was born on March 13, 1911. When he was about two his family moved to Montana where he learned to be a rough western cowboy. His mother was thoroughly educated woman. She taught him how to read and write at an early age. L. Ron Hubbard was said to have an avid interest in life at an early age. When his father’s naval career required that they move away from Montana, his mother was responsible to teach him the schooling he missed. When Ron was twelve the family moved to Seattle, Washington. Ron got the liberty to be associated with many different types of people such as the Blackfoot Indians, Beijing magician, Chinese magicians, and studying Buddhists. He enrolled at George Washington University, his father put him into mathematics and engineering. Now he learned how to take the scientific approach to solving problems. He became more and more interested in the way the mind works. He would ask the professors at George Washington University and they could not give him an answer, so he came up with the idea that the western culture did not understand a thing about the mind. He believed that it was his duty to explore the mind and to tell the western world how it works. This is when he came up with dianetics and then eventually the religion of scientology. Scientology consists of eight dynamics. With these dynamics, a person realizes that his life extends beyond himself. The first dynamic is â€Å"self.† This teaches you to survive as an individual. Taking care of such thi... ... deal with life here on earth and teaches people how to become better people. Improving conditions here on earth is the foremost important thing to fix. Scientology seems to address this issue with full force. It uses modern technology, and knowledge of how human relations to improve the quality of life of people who chose to follow Scientology. In the description of L. Ron Hubbard, I was disgusted. They made it sound like he was this extremely smart guy. When he started studying the mind and the way it works, he noticed that the western society knew nothing about it. He sounded pompous when he said â€Å"it was very obvious that I was dealing with and living in a culture which knew less about the mind than the lowest primitive tribe I had ever come in contact with†¦Ã¢â‚¬  I did not like this at all. He sounds like a very arrogant man. Arrogant in a cocky way. In doing research I noticed that all of his followers seem to have the same attitude. I find that it is funny that they are so dedicated to following a man who was born in the twentieth-century. He is an ordinary man who happened to be exceptionally smart. He is a business man who is good at persuading people and wants to make money.