Effect On The British Invasion Bands History Essay

 

Throughout history, there have been a great number of instances where certain aspects of the time period had an effect on art forms of the time. A good example of this is how the decade the 1960’s affected the British Invasion rock and roll groups. There were a great number of historical events that occurred during the 1960’s, both positive and negative. One event that had already been taking place for five years before the decade even began was the Vietnam War, which was a violent conflict that pitted the United States and South Vietnam against the Communists of North Vietnam. The war began due to North Vietnam’s efforts to unite itself with South Vietnam, thus making South Vietnam Communist. The United States did not want Communism to spread in Asia any more than it already had, so eventually the United States entered the war. The war was unsuccessful, however, since the United States troops were not prepared for the Vietnamese climate conditions and the guerrilla warfare tactics of the North Vietnamese.

While these major historical events were going on, rock and roll had just begun to take off. Although rock and roll didn’t truly reach its creative peak until the 1960’s, the genre actually grew and became popular in the 1950’s. Rock and roll in itself has been thought by some to be simply a combination of country music and rhythm and blues. This is not the case, however, as the original ideas of rock and roll were in place for a long time, although it did not become a whole separate genre until the mid 1950’s. Probably the biggest influence on how rock and roll became so popular is Elvis Presley. Following his influence, artists such as Little Richard, Buddy Holly, and Chuck Berry started what was called rock and roll’s golden era. It wasn’t until the mid 1960’s that rock and roll had another wave of change comparable to this one.

The British Invasion was a musical movement, which consisted of English rock and roll bands that found fame in both the United Kingdom and the United States. The British Invasion bands did not receive recognition right away, however. In fact, many bands attempted to copy the American style of rock in the 1950’s, although many had little success. The reason for this is that they were unable to grasp the rock and roll feel for the most part. By 1962, some of Britain’s youth was able to understand the rock genre and a few bands became popular in Liverpool, especially the Beatles. It was the Beatles, in fact, who was the first band to go to the United States and spread their own type of rock overseas. After their example, many bands followed. By 1964, Britain had become the birthplace of such popular bands as The Who, The Rolling Stones, The Yardbirds, and others. (Britannica.com – British Invasion) Between 1964 and 1966, the British Invasion bands had successfully brought their brand of music over to the United States and had numerous hits including Downtown by Petula Clark, Do Wah Diddy Diddy by Manfred Mann, Wild Thing by the Troggs, Satisfaction by the Rolling Stones, and many others. (Britannica.com – British Invasion)

Since the 1960’s was a time period in which sex, love, violence, and drugs were running rampant, it is evident that some of these social factors of the time period had an effect on these bands. The Beatles and the Rolling Stones were two of the most popular British Invasion bands in both the United States and The United Kingdom at the time. They are also prime examples of social factors have affected the music of the time period. The Vietnam War, which was directly related to the Hippie Movement, was one factor that had an effect. Mass drug use of the time period was also a factor that had a major influence on the British Invasion bands. Besides all of that, the media also had a tremendous effect on many of the bands.

The Vietnam War was received with mixed feelings in the United States, and many people’s opinions were negative. Although some people were for the Vietnamese struggle, many felt that the United States would not have the ability to win and believed that it was not the United States responsibility to fight for South Vietnam. One group of people in the United States, most generally called “Hippies,” felt that violence in any form should not be present and also advocated environmental issues and love. Hippies also usually dressed in clothing with bright colors with flowers printed on them. They also tended to wear headbands and put flowers in their hair.

The hippie movement had an effect on the music of the time in which many of the British Invasion bands wrote songs that included many of the hippie ideas. For example, in the song, In Another Land by the Rolling Stones, the lyrics read, “In another land where the breeze and the trees and flowers were blue I stood and held your hand. And the grass grew high and the feathers floated by. I stood and held your hand. And nobody else's hand will ever do. Nobody else will do.” (The Rolling Stones Albums) These lyrics show that the Rolling Stones showed feelings for nature and the environment, as well as love, which directly relates to the hippie style.

Besides the musical influence the hippies had on the British Invasion bands, there was also a fashion influence as well. In the most of the bands it was typical for most of the members to have fairly long hair or dress in a hippie manner. An example of this is throughout most of the Beatles career in the 1960’s, they had slightly long hair in the front, which was fairly consistent with the hippie style. Even though they began their career wearing matching suits, they later on wore clothing that sported more of a hippie influence.

Although the hippie movement played a major part in having an effect on the British Invasion bands, another major influence on many of the bands, which was also related to the hippie movement were the use of psychoactive drugs such as LSD (lysergic acid diethylamide) and marijuana. LSD is a hallucinogen, which basically means that when taken, it puts the user in an altered state of consciousness. The sensations first occur about forty minutes after the drug is taken. After about an hour, the intensity level of the drug is at its highest and lasts for about four to five hours. People who have experienced LSD have said that during that peak point that their senses became stronger, and also sometimes seemed to mix. Also, while on the drug it appears as if inanimate objects can move and bend. (Psychological effects of LSD) It is also said that LSD can give the user a new dimension of creativity, which resulted in the use of the drug by many rock bands in order to come up with fresh, new ideas. Another psychoactive drug, marijuana is a plant that contains the chemical THC (tetrahydrocannabinol). THC is what gives marijuana its mind-altering control and gives what many people call, a “high”. Most often marijuana is smoked, although it can also be eaten. Marijuana also is said to help people think more creatively.

Perhaps two of the most popular bands who had changed their musical style through the use of psychoactive drugs are the Rolling Stones and the Beatles. The birth of psychedelic rock is what brought about this change in these bands. Psychedelic rock began in the late 1960’s and was the result of LSD use as well as feedback and electronic sounds. The most popular psychedelic rock band was the Grateful Dead, who actively used drugs during their concerts. (Britannica.com – Psychedelic Rock) The Rolling Stones and the Beatles both became influenced by this type of rock, and wrote songs that showed many signs of drug use. Between 1966 and 1967, The Beatles created albums such as Revolver, Sgt. Pepper's Lonely Hearts Club Band, and Magical Mystery Tour. These albums all had psychedelic influences in them. For example, in the song Magical Mystery Tour, the lyrics are, “Roll up, roll up for the Mystery Tour. Roll up, and that’s an invitation. Roll up, to make a reservation. Roll up for the Mystery Tour. The Magical Mystery Tour is waiting to take you away, waiting to take you away.” (The Beatles Discography – US Albums) It is evident that based on these lyrics the “magical mystery tour” is a marijuana high because the term “roll up” refers to rolling up a marijuana joint, and the “magical mystery tour” as well as “taking you away” refer to getting high. This is a major difference from the lyrics in earlier songs the Beatles had written.

The Rolling Stones also experimented with psychoactive drugs, which in turn affected their music. The songs in their album, Their Satanic Majesties Request, displayed a psychedelic side. In the song, Two Thousand Light Years From Home, the lyrics read, ”Sun turnin' 'round with graceful motion. We're setting off with soft explosion. Bound for a star with fiery oceans, it's so very lonely, you're a hundred light years from home.” (The Rolling Stones Albums) These lyrics have a very fantasy-like manner to them, in which speaking of “fiery oceans” and “soft explosions” hint that they are the creative result of an LSD trip. Also, saying, “you’re a hundred light years from home” may have a hidden meaning of being high.

Though not directly related to the hippie movement or drugs, the media had an extremely large influence on the British Invasion bands. Television had reached an extremely high popularity by the 1960’s and therefore provided a new way for the bands to gain exposure rather than just live events, or even word from newspapers or magazines. Even though the 1960’s took place well before the MTV era, bands were still seen on television shows such as “The Ed Sullivan Show” and “The Tonight Show with Johnny Carson.” It was on these television appearances that the Beatles performed that truly started the British Invasion. Since the Beatles had instantly gained incredible amounts of popularity in the United States because of these television appearances, many other bands seized the opportunity and presented themselves to the American audience.

Media exposure also had a negative influence on some of these bands, however. For example, in an interview in the London Evening Standard, John Lennon of the Beatles said, “Christianity will go. It will vanish and shrink. I needn't argue with that; I'm right and I will be proved right. We're more popular than Jesus now; I don't know which will go first - rock 'n' roll or Christianity. Jesus was all right but his disciples were thick and ordinary. It's them twisting it that ruins it for me.” (The Dark Side of Beatlemania – John Lennon) At that time it was not considered to be anything negative. Five months later however, Datebook, a United States magazine for teenagers reprinted that quote and featured it in an article titled, “The Ten Adults You Dig/Hate The Most”. Since the quote had been misconstrued by the media, many people in the United States interpreted it as though the Beatles were antichrists. Radio stations stopped playing Beatles’ music on the air and many children around the country publicly destroyed Beatles records, memorabilia, etc. Other bands quickly learned that the media could easily twist and turn what one says, so they would have to be more careful.

In conclusion, based on these main points it is evident that the 1960’s truly did have a huge impact on the British Invasion bands. The marijuana and LSD use by many of the bands gave their music a fresh, new sound that had never been heard before. Also, the hippie influences added a new way for social concerns to be expressed. Finally, the British Invasion bands also learned that the media can portray them in both positive and negative ways, based on how they manipulate words or actions.

After thoroughly researching the 1960’s as well as the bands that made up the British Invasion, I feel that the sixties was one of the most eventful, tragic, interesting, and beautiful time periods in world history. The stories from the two major conflicts going on – The Vietnam War, as well as the struggle back in the United States between the war supporters and those against it are absolutely riveting. I have also found that there are tremendous similarities between the hippies and the authors of the Romantic Era in Britain. Both the hippies and the romantics felt very strongly about nature and expressed their feelings in art forms, the romantics through poetry and the hippies through song. I also have found similarities between psychedelic rock and the works of Samuel Taylor Coleridge, who wrote works such as Kubla Kahn and The Rime of the Ancient Mariner. Because of Coleridge’s use of Opium, his works showed signs of fantasy, very much like the psychedelic style. I feel that not only has the 1960’s had a major effect on the British Invasion bands, but I also feel that the bands themselves have drilled a deep impact into the culture of the 1960’s as well as today.

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Study The Effect Of Spoilers On Wings Engineering Essay

 

While travelling from an aircraft it must have been observed many a times that during a flight and after touchdown of aircraft a part of the wing in between the leading and trailing edges of the wing is deployed. This part of the wing is called the spoilers. Especially after the touchdown, it can clearly be observed that the spoilers are extended upward into the airflow.

Spoilers as the name itself explains, it means to spoil. The spoilers, spoils the airflow over a wing and decreases the lift of an aircraft. Here by spoiling the airflow means to disturb the airflow and decreasing the lift by increasing the drag. Spoilers can be used to slow an aircraft, or to make an aircraft descend, if they are deployed on both wings. Spoilers are also used to generate a rolling motion for an aircraft, if they are deployed on only one wing. Spoilers are used for multipurpose; they are sometimes used when descending from cruise altitudes to assist the aircraft in descending to lower altitudes without picking up speed.

An increased rate of descent could also be achieved by lowering the nose of an aircraft, but this would result in an excessive landing speed. However, when spoilers are used along with the thrust reversers they also help in considerably decreasing the runway distance required by an aircraft to land safely and enable the approach to be made at a safe speed for landing. In a descent without spoilers, air speed is increased and the engine will be at low power, producing less heat than normal. The engine may cool too rapidly, resulting in stuck valves, cracked cylinders or other problems. Spoilers alleviate the situation by allowing the aircraft to descend at a desired rate while letting the engine run at a power setting that keeps it from cooling too quickly.

Spoilers are hinged, rectangular plate-like structures installed flush along the top of an aircraft wing, just forward of the flaps. When the pilot activates the spoilers, the plates pivot up on their center hinge fittings into the airstream. By doing so, the spoiler creates a carefully controlled stall over the portion of the wing behind it, greatly reducing the lift of that wing section, as a result, the airflow over the wing is disturbed (spoiled) and lift is decreased with the increment in the drag. This can be observed from fig.1 below.

Fig. 2.1: Spoilers on a wing.

The spoilers works in different conditions of the flight as per desirable by the pilot. All these conditions are discussed in the underwritten sections of the report.

On landing, however, the spoilers are nearly always used at full effect to assist in slowing the aircraft. The increase in form drag created by the spoilers directly assists the braking effect. However, the real gain comes as the spoilers cause a dramatic loss of lift and hence the weight of the aircraft is transferred from the wings to the undercarriage, allowing the wheels to be mechanically braked with much less chance of skidding.

Fig.2.2: Spoilers in action

In the above Fig.1.2 we can clearly observe the disturbance i.e. the turbulence that is caused in the flow when the spoilers are deployed. This turbulence which is created in the flow is the major cause for decrement in lift along with the increase in the drag.

The spoilers may also be differentially operated to provide roll control. On landing, however, the spoilers are nearly always used at full effect to assist in slowing the aircraft. The increase in form drag created by the spoilers directly assists the braking effect. However, the real gain comes as the spoilers cause a dramatic loss of lift and hence the weight of the aircraft is transferred from the wings to the undercarriage, allowing the wheels to be mechanically braked with much less chance of skidding. Reverse thrust is also often used to help slow the aircraft on landing. The spoilers may also be differentially operated to provide roll control.

The spoilers are used in multiple conditions such as:

When the pilot deploys the spoilers, the plates flip up into the air stream. The flow over the wing is disturbed by the spoiler, the drag acting on the wing is increased, and thus as a result lift is decreased. During the mid air i.e. when an aircraft is airborne and spoilers are deployed by the pilot the aircraft descend from cruise altitudes to assist the aircraft in descending to lower altitudes without picking up speed. This is very helpful in decreasing the altitude of the aircraft, without the use of propulsive or any other power.

Fig.3.1: Spoiler up position.

On landing, however, the spoilers are nearly always used at full effect to assist in slowing down the aircraft. The increase in form drag created by the spoilers directly assists the braking effect. However, a major advantage is that the spoilers cause a dramatic loss of lift and hence the weight of the aircraft is transferred from the wings to the undercarriage, allowing the wheels to be mechanically braked with much less chance of skidding. By the use of spoilers at the time of landing after touchdown gives efficiency to the brakes. Reverse thrust is also often used to help slow the aircraft on landing along with the spoilers (consider Fig.3.2).

By use of spoilers along with thrust reversers effectively stops the aircraft on landing and also helps in reducing the required ground distance for landing.

Fig.3.2: Spoilers being used after touchdown.

They are useful on gliders to vary the lift-to-drag ratio for altitude control and on airliners on landing to reduce lift quickly to prevent the airplane from bouncing into the air. During the time of landing the aircrafts needs to have least lift, if there is a little misbalance and lift is produced on the wing then instead of landing the aircraft will bounce back in the air. To avoid this situation spoilers are very helpful in dumping the lift acting on the aircraft.

A single spoiler is used to bank the aircraft; to cause one wing tip to move up and the other wing tip to move down. The banking creates an unbalanced side force component of the wing lift force which causes the aircraft's flight path to curve.

Fig.3.3: Roll motion caused by spoilers.

If the airplane's right wing spoiler is deployed, while the left wing spoiler is stored flat against the wing surface (as viewed from the rear end of airplane) consider Fig.3.3. The flow over the right wing will be disturbed by the spoiler, the drag of this wing will be increased, and the lift will decrease relative to the left wing. The lift force is applied at the center of pressure of the segment of the wing containing the spoiler which in result creates a torque about the center of gravity. The net torque causes the aircraft to rotate about its center of gravity.

The resulting motion will roll the aircraft to the right (clockwise) as viewed from the rear. If the pilot reverses the spoiler deflections (right spoiler flat and left spoiler up) the aircraft will roll in the opposite direction.

The aircraft rolls because one aileron is deflected downward while the other is deflected upward. Lift increases on the wing with the downward-deflected aileron because the deflection effectively increases the camber of that portion of the wing. Conversely, lift decreases on the wing with the upward-deflected aileron since the camber is decreased. The result of this difference in lift is that the wing with more lift rolls upward to create the desired rolling motion. Now, Consider Fig.3.4.

Fig.3.4: Adverse Yaw.

Unfortunately, drag is also affected by this aileron deflection. The induced drag and profile drag, are increased when ailerons are deployed. Thus, the wing on which the aileron is deflected downward to generate more lift also experiences more induced drag than the other wing. The profile drag increases on both wings when the ailerons are deflected, but the increase is equal. However, the induced drag on each side is not equal, and a larger total drag force exists on the wing with the down aileron. This difference in drag creates a yawing motion in the opposite direction of the roll. Since the yaw motion partially counteracts the desired roll motion, we call this effect adverse yaw.

When used in coordination with ailerons, a spoiler can be used to reduce the lift and increase the profile drag on the wing with the up aileron. As a result, the wing with the down aileron experiences a large increase in lift and a small increase in drag while the wing with the up aileron experiences a large decrease in lift and a large increase in drag. These effects combine to create the desired roll motion and a complimenting yaw motion that is called proverse yaw.

By the use of spoilers a rapid descents may be made without having to reduce power, thereby maintaining engine temperatures at a comfortable level, and eliminating the risk of engine "shock cooling." In time of a descent without spoilers, i.e. by simply reducing the throttle the air speed is increased and the engine will be at low power, producing less heat than in normal. Thus as a result the engine may cool too rapidly, resulting problems such as, stuck valves, cracked cylinders or other problems. Spoilers alleviate the situation by allowing the aircraft to descend at a desired rate while letting the engine to run at a power setting that keeps it from cooling too quickly.

Wood is used to make the solid wing as per the coordinates; Steel plate is used to show the spoiler on the wing, nuts and bolts.

The experiment was conducted in the aerodynamics lab at the university. Wind tunnel is used for the testing of solid wing. This is a low speed and low turbulence wind tunnel. The experiment was performed at tunnel velocity of 30m/s. Lift and drag on the solid wing was calculated before and after the spoilers were deployed.

A NACA 0015 symmetrical airfoil with a 25% thickness to chord ratio was analysed to determine the lift and drag. The chord of the airfoil is 15 cm; using the NACA 0015 symmetric airfoil coordinates the solid wing was made. The coordinates of NACA 0015 are:

1

0.95

0.9

0.8

0.7

0.6

0.5

0.4

0.3

0.2

0.15

0.1

0.075

0.05

0.025

0.0125

0

0.0125

0.025

0.05

0.075

0.1

0.15

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0.95

1

0.0012

0.01027

0.01867

0.0332

0.0448

0.0532

0.05827

0.06

0.05827

0.05293

0.04867

0.0424

0.03813

0.03267

0.02453

0.01813

0

-0.01813

-0.02453

-0.03267

-0.03813

-0.0424

-0.04867

-0.05293

-0.05827

-0.06

-0.05827

-0.0532

-0.0448

-0.0332

-0.01867

-0.01027

-0.0012

Fig.4.1: NACA 0015 airfoil

The wind tunnel testing of solid wing was performed, first with normal condition i.e. when spoilers are not deployed. The following results are:

1

0°

0.164 N

0.377 N

2

5°

20.23 N

0.725 N

3

10°

33.02 N

1.54 N

4

15°

16.2 N

6.525 N

Now, for the second case i.e. when the spoiler is deployed. The values obtained are:

1

0°

-0.034 N

0.549 N

2

5°

12.94 N

6.45 N

3

10°

24.05 N

13.37 N

4

15°

9.85 N

20.58 N

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Effect Of Counterfeit Branding On Sales Of Edhardy Marketing Essay

This research proposal is objectively aimed at understanding all the hurdles currently experienced by Ed Hardy in expanding their business in India. The business organization with intent to expand in different market entered India in the year 2007 as part of its expansionist policy to explore the commercial viability of the market objectively aiming at high profitability (Khilnani 2009).

In the year 2002, Ed Hardy who initially commenced his career as a tattoo artist entered into an agreement with Ku USA, Inc. to carry out and market a clothing line with the basis of his tattoo art design which was extraordinarily popular in the U.S.A. In a span of just two years, the international corporate giant Saks expressed interest in Ed Hardy clothing line. Hardy and Ku USA created the Hardy Life LLC, an organization possessing trademark ownership in addition to copyrights to all of Ed Hardy’s images.

Christian Audigier entered the Ed Hardy scenario in 2004 gaining rights for the clothing line production based on the artistic designs of Ed Hardy. Ed Hardy in today’s day and age has a commanding global position with outlets in New York, Los Angeles, Seattle, Honolulu, the Gulf Nations and India.

Iconic Brand Group made an acquisition of 50% of shares in Hardy Way, LLc. Iconic Brand Group made a payment of $ 17 million for major share in the Hardy Way as it estimates sales revenue to be to the tune of approximately £110 million by 2011.

Ed Hardy is undoubted recognized as an organization of high repute with its product lines having immense popularity especially among the younger generation. In European nations such as the U.K., U.S, Germany and France, the organization commands a high market share as its varied line of products such as t-shirts, shirts, shoes, belts and deodorants are highly consumed by the consumers. The organization has branches globally and has recently set up branches in India. (Chaudhari 2009)

India as a market is attracting a great number of Multinational as the high populace makes provision for a ready market for the merchandise dispatched for sale. Though Ed Hardy has numerous outlets at strategic consumer locations in India especially Mumbai that is the capital centre of the country, the business is primarily plagues with issues relating to counterfeiting. (Kotler 2007)

This decision for conduct of this research was essentially taken to highlight the issues faced by Ed Hardy. This brand has been a victim to Counterfeit Branding. Counterfeit Branding is highly prevalent in countries like India and China and these results in economic losses to the respective governments by way of taxes and the business are affected in terms of profitability. Ed Hardy T-shirts which are a rage amongst the college-going consumer and young adults commence at a pricing range of approximately Rs 1000 (20 USD) which is relatively on the higher side in terms of pricing. The pricing, though is not directly a contributing factor for the declining sales of the brand, counterfeiting of Ed Hardy products cause a massive impact on the sales and consequently the revenue (Keller 2007).

http://www.bestshoeswomen.com/images_products/Ed_Hardy_Original_Lowrise_100_Women_Shoes_Sneakers.jpg http://www.celebrityclothingline.com/wordpress/wp-content/uploads/2009/12/Ed-Hardy-Tshirt-Beautiful-Ghost-Graphic-Original-62-4499.JPG http://eleven.se/files/w/wallet-black-1_250x250.jpg http://www.funkytrend.com/wp-content/uploads/2009/02/ed-hardy-true-love-belt-thumb.jpg http://www.clothingcheapprice.com/images_products/Ed_Hardy_Emerald_City_Elphaba_Tote.jpg http://www.newwavefragrances.com/images_up/1239994301_art_EH-M-LINESHOT-450x400.jpg

Comparative Analysis of the Ed Hardy’s original and counterfeit products: http://www.revolveclothing.com/Brands.jsp?c=Ed+Hardy&sc=Shoes

Ed Hardy Product Line

Original

£ (Pounds)

Counterfeit

£ (Pounds)

Pricing Difference

£ (Pounds) (Approximately)

1

T-Shirts

26

6

20

2

Shirts

31

2

29

3

Converse Shoes/Heels

65

12

53

4

Belts

25

5

20

5

Wallets/Purses

60

7

53

6

Handbags

265

25

240

7

Perfumes

109

10

99

Ed Hardy commenced its retail chain at Mumbai, India in 2007 and has in continuity reported declining sales for the last three financial years citing counterfeiting issues as the primary reason for the failure of the brand to succeed in the Indian market. There is a huge pricing difference between the original and counterfeit product leading prospective consumers to continue the purchase of counterfeit products. The counterfeit products are reported almost similar to the original in terms of design, dimensions and labeling but lacks quality of the original yet it this factor does not prove to be discouraging for consumer who continue to patronize counterfeit Ed Hardy products. Ed Hardy, Nike, Rolex, Louis Vuitton, Dior, Yves St Laurent, Prada, Hermes and Cartier are generally the brands that are highly counterfeited. These brands have not reported higher profits in the last several years. (Simon 2005)

Year

Sales Revenue (Pounds)

1

2007

40 million pounds

2

2008

28 million pounds

3

2009

10 million pounds

The above table is clearly indicative of a constant and steady decline in the sales of the Ed Hardy Mumbai Outlet wherein at the first year of establishment of the outlet the reported sales was at 40 million pounds which declined to 28 million pounds and finally to 10 million pounds in the accounting year of 2009 (Kumar 2009).

Anti-counterfeiting laws and legislations though in existence have not been strictly imposed. The ACTA Act which is an Anti-Counterfeiting Trade Agreement Act between global trading nations of the world such as U.S.A. , U.K., Japan, France, India and many Asian countries has been ineffective in reducing if not totally eliminating the production and sale of counterfeit products. Counterfeit products in India are a booming industry which is crippling the economy and business ventures of especially fashion brands such as Ed Hardy. The Indian Judicial System is not as stringent and enforceable as the European counterparts. Strict punitive action is not taken against individual or firms indulging in counterfeiting reputed brands and hence this activity continues to flourish cause huge financial losses for organizations such as Ed Hardy and many more that have invested highly in the Indian market in terms of setting up retail outlets, promotion and advertising and all related expenditure. (Hakan 2009)

At times there have been recorded cases of legal action taken against offenders who are let off after an imposition of a penalty charge which is very minimalistic defeating the very purpose of the exercise. (Nayadu 2009)

India is generally characterized as a cost conscious economy with the general tendency of the consumers to buy or patronize products which are specifically low on cost. Quality of the product is a second consideration after the pricing factor of the product. Hence, international fashion brands such as Ed Hardy target a certain market segment which is the upper middle class wherein affordability of the product does not pose a problematic issue. This results in a narrowed target market segment. This segment would have issues in buying the high priced Ed Hardy product but with counterfeit Ed Hardy products flooding the market, the credibility of the original product is at stake as its exclusiveness is lost in the market of this product. The consumers that cannot afford the original product line of Ed Hardy turn to counterfeit substitutes to gain satisfaction of possessing the product and this has a negative effect on the limited market segment consumers of Ed Hardy as the novelty of the product is lost. (Puri 2009)

Besides high level counterfeiting of Ed Hardy product, the business House faces severe competition from other popular and well established brands such as Levis, Spykar, Adidas, Nike, Louis Phillip and many more that entered the Indian Market several years ago and have successfully gained good market prominence. These brands are well aware of the price conscious mentality of the Indian consumer and constantly indulge in competitive pricing strategies and promotional techniques to increase sales and subsequent profitability. Ed Hardy being relatively new in the Indian Market Scenario is hence confronted with hurdles in this form of excess competition thereby retarding growth and progress in the process. (Armstrong 2005)

The Government of India taxation policies toward FDI (Foreign Direct Investment) especially of luxury brands is comparatively and competitively high thereby creating a need for such multinational business organizations such as Ed Hardy to have higher sales to sustain themselves in the Indian market and grow simultaneously. The taxation rates of such brands amount to 40% of the product price which is ultimately borne by the consumer. Such taxation policies towards fashion brands are imposed with the intention to protect local and national entrepreneurs and their businesses form highly popular reputed brands such as Ed Hardy that command a high percentage of clientele globally.

This taxation policy of the Government of India has proven to be a deterrent factor contributing to slack in growth for Ed Hardy. (Keegan 2002)

To understand the obstacles faced by Ed Hardy and strategize ways to counter attack these problems

To identify the hurdles confronted by the Fashion Brand Ed Hardy in business expansion in the Indian Market

To ascertain if the business house been in a progressive state post-commencement in India

To establish what the future business prospects of the organization are

To gain knowledge if the Indian business market scene too competitive for the survival of Ed Hardy

To analyze if the Indian business market scene too competitive for the survival of Ed Hardy

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The Effect Of Three Antiseptics On Staphylococcus Aureus Biology Essay

Biology » The Effect Of Three Antiseptics On Staphylococcus Aureus Biology Essay

With food and air- borne infections rising by the day, skin infections are the most commonly known infections. Staphylococcus aureus bacteria generally cause skin infections that lead to various diseases. Antiseptics are well known for preventing the bacteria from infecting the skin. Therefore in my experiment I would like to conduct a check on “The effect of three antiseptics on Staphylococcus aureus.”, keeping the time for the antiseptic to act on the surface constant but varying the dilutions to check the effect on the rate of bacterial growth on the surface. This experiment has been proved successful and is termed as the microbial challenge test.

Antiseptics and disinfectants are used extensively in hospitals and other health care settings for a variety of topical and hard-surface applications. In particular, they are an essential part of infection control practices and aid in the prevention of nosocomial infections. A wide variety of active chemical agents (or “biocides”) are found in these products, many of which have been used for hundreds of years for antisepsis, disinfection, and preservation. “Biocide” is a general term describing a chemical agent, usually broad spectrum that inactivates microorganisms. Because biocides range in antimicrobial activity, other terms may be more specific, including “-static,” referring to agents which inhibit growth (e.g., bacteriostatic, fungistatic, and sporistatic) and “-cidal,” referring to agents which kill the target organism (e.g., sporicidal, virucidal, and bactericidal).

Types of Antiseptics

Chlorhexidine Gluconate

A biguanidine derivative, used in concentrations of 0.5–4.0% alone or in lower concentrations in combination with other compounds, such as alcohols. Used as a skin antiseptic and to treat inflammation of the gums (gingivitis). The microbicidal action is somewhat slow, but remanent. It is a cationic surfactant, similar to Quats.

Alcohols

Most commonly used are ethanol (60–90%), 1-propanol (60–70%) and 2-propanol/isopropanol (70–80%) or mixtures of these alcohols. They are commonly referred to as "surgical alcohol". Used to disinfect the skin before injections are given, often along with iodine (tincture of iodine) or some cationic surfactants (benzalkonium chloride 0.05–0.5%, chlorhexidine 0.2–4.0% or octenidine dihydrochloride 0.1–2.0%).

Quaternary ammonium compounds

Also known as Quats or QAC's, include the chemicals benzalkonium chloride (BAC), cetyl trimethylammonium bromide (CTMB), cetylpyridinium chloride (Cetrim, CPC) and benzethonium chloride (BZT). Benzalkonium chloride is used in some pre-operative skin disinfectants (conc. 0.05–0.5%) and antiseptic towels. The antimicrobial activity of Quats is inactivated by anionic surfactants, include chlorhexidine andoctenidine.

Infection: Before a microbe or parasite can invade the host and cause infection, it must first attach to and penetrate the surface of the epithelial layers of the body. Organisms gain entrance into the body through active or passive means. That is they have to pass physical barriers such as the skin or epithelial cells which line the mucosal surfaces of the respiratory, gastro intestinal and genitourinary tracts. The active passage for these microbes and parasites are the dead layers of the skin.

Skin: The term ‘skin’ applies to the outer covering of vertebrate animals. The skin is the largest organ of the body and has many different functions. The structure of the skin varies in different vertebrate groups. The basic structure of human skin will be described here. The skin is composed of two main layers, the epidermis and dermis. These cover the underlying, or ‘subcutaneous’ tissue, which contains specialised fat- containing cell known as adipose tissue. The thickness of the epidermis and dermis varies according to the region of the body and from person to person.

Epidermis- The cells of this region are separated from the dermis by a basement membrane. The epidermis is composed of many layers of cells that form a stratified epithelium. The cells immediately above the basement membrane are cuboidal epithelial cells and form a region known as the Malpighian layer. The cells in this layer are constantly dividing by mitosis and replace all the cells of the epidermis as they wear away. The Malpighian layer forms the lower region of the stratum granulosum, which is composed of living cells that become flatter as they approach the outer region of the epidermis, the stratum corneum. Cells in this region become progressively flattened and synthesise keratin, which is a fibrous protein which makes the cells waterproof. As the keratin content of the cells increases, they are said to become ‘cornified’. Their nuclei disappear and the cells die. The thickness of the stratum corneum increases in part of the body where there is considerable friction, such as the ball of the foot and the bases of the fingers. The outer covering of the skin forms a semi- transparent, thin, tough, flexible, waterproof covering by the hair follicles and by pores, which are the pores, which are the openings of the sweat glands. The outermost squamos epithelial cells are continually being shed as a result of friction.

The stratum corneum has become modified in many vertebrates to produce nails, claws, hooves, horns, antlers, scales, feathers and hair. Keratin is the main component of all these structures.

Dermis- The dermis is a dense matrix composed of connective tissue rich in elastic fibres and contains blood capillaries, lymph vessels, muscle fibres, sensory cells, nerve fibres, pigment cells, sweat glands and hair follicles.

The skin also has other more specific immune functions. Langerhan cells, which are immunocompetent cells, are involved in recognizing and processing foreign substances. These decrease with age, altering immune function (Gilchrest, 1999).

Mucous membranes, covering our eyes, alimentary canal, and genito-urinary tracts, play a protective role in providing a barrier and differential absorption. Usually, the skin prevents invasion by microorganisms unless it is damaged—for example, by an injury, insect bite, or burn.

Staphylococcus Aureus: Out of the many bacteria entering the human body through the dead skin, Staphylococcus aureus, is the most potent organism infecting the skin.

Facultatively anaerobic - do not require oxygen for growth but do grow better in its presence

The most common species of staphylococcus is Staphylococcus aureus which is a pathogen of both humans and animals. It is characterized by its ability to clot blood plasma by action of the enzyme coagulase

a) Colony Characteristics of Staphylococcus aureus

Size - 1 micrometre

Gram nature - Gram positive

Appearance- Grape-like cluster but some occur as single or pair of cells

usually non-capsulate

Form - Colonies are circular, 2-3 nm in diameter with a smooth shiny surface when grown on nutrient agar, milk agar or blood agar for 24 h at 37 degrees

Pigmentation - Colonies are often pigmented, though a few strains are unpigmented

Halophile i.e.Salt-tolerant

b) Pathogenesis of Staphylococcus aureus

Present in the nose of 30% of healthy people and may be found on the skin, causes infection at sites of lowered host resistance.

Eg: - damaged skin or mucous membranes

c) Epidemiology of Staphylococcus aureus

i) Infected Lesions:

Large no. of staphylococci are disseminated (spread widely) in pus and dried exudates discharged from large infected wounds, burns, secondary infected skin lesions and in sputum coughed from the lung of a patient with bronchopneumonia. Direct contact is the most important mode of spread, but air-borne dissemination may also occur

ii) Healthy Carriers:

Spread into environment by the hands, handkerchiefs, clothing and dust consisting of skin squames? and cloth fibres of healthy carriers. Some carriers, called shedders, disseminate exceptionally large no. of cocci, comparable to the number disseminated by patients with large superficial lesions or lower respiratory tract infections

iii) Animals:

Domesticated and some wild species may disseminate Staphylococcus aureus from infected lesions or carriage sites and so cause infections in humans.

Mode of infection may be either exogenous (from an external source) or endogenous (from a carriage site or minor lesion, elsewhere in the patient's own body). Staphylococci do not grow outside the body except occasionally in moist nutrient materials such as meat, milk and dirty water. Although not spore-forming, they may remain alive in a dormant state for several months when dried in pus, sputum, bed clothes or dust

They are fairly readily killed by heat (65 degrees for 30 min), by exposure to light and by common antiseptics. Therefore I would like to study on the research question,

Resistance to antimicrobial agents (AMR) has resulted in morbidity and mortality from treatment failures and increased health care costs. Appropriate antimicrobial antiseptic use has unquestionable benefit, but physicians and the public frequently use these agents inappropriately.

The microbial challenge test would be most appropriate to study the effect of antiseptics on Staphylococcus bacteria.

Microbiological challenge testing has been and continues to be a useful tool for determining the ability of a product to prevent the growth of pathogens. Microbiological challenge tests also play an important role in the validation of processes that are intended to deliver some degree of lethality against a target organism or group of target organisms.

The design, implementation, and assessment of microbiological challenge studies is a complex task that depends on factors related to how the product is formulated, manufactured, packaged, distributed, prepared, and used.

When conducting a microbiological challenge study, a number of factors must be considered.

These include:

(1) the selection of appropriate pathogens,

(2) the level of challenge inoculum,

(3) the inoculum preparation and method of inoculation,

(4) the duration of the study,

(5) formulation factors and storage conditions,

(6) sample analyses.

The interpretation of the data and pass/fail criteria are critical in evaluating whether a product needs time/temperature control for safety.

It is also important to incubate and prepare the challenge suspension under standardized conditions and format. Shifts in the incubation temperature used to propagate the challenge organisms and the storage temperature of the product have been shown to change the length of the lag period of the challenge study itself.

The inoculum level used in the microbiological challenge study depends on whether the objective of the study is to determine product stability and shelf life or to validate a step in the process designed to reduce microbial numbers.

The preparation of the inoculum to be used in microbiological challenge testing is an important component of the overall protocol. Typically, for vegetative cells, 18 to 24 h cultures revived from refrigerated broth cultures or slants or from cultures frozen in glycerol are used. The challenge cultures should be grown in media and under conditions suitable for optimal growth of the specific challenge culture.

The method of inoculation is another extremely important consideration when conducting a microbiological challenge study. Every effort must be made not to change the critical parameters of the product formulation undergoing challenge. There are a variety of inoculation methods that can be used depending upon the type of product being challenged.

Standard suspensions of insoluble Barium sulphate precipitates have been described by Mc. Farland (1907) and Brown (1919-20). A series of standards of different BaSO4 concentrations correspond to suspensions of different numbers of bacteria/ml.

Preparation of Mc. Farland standards.

Std. Tube no.

BaCl2 1% (ml)

H­2SO4 1% (ml)

Corresponding bacterial concentrations (millions/ml)

0.5

0.05

9.95

150

1

0.1

9.9

300

2

0.2

9.8

600

3

0.3

9.7

900

4

0.4

9.6

1200

5

0.5

9.5

1500

Use:

In a tube of the same internal diameter as the standard, prepare a uniform suspension in saline of bacteria under test to a density greater than that required.

Select a standard opacity tube of the density required and shake it vigorously until all the deposit is raised into uniform suspension.

At once compare the bacterial suspension with the standard. View with oblique illumination against a dark background or over a printed page.

Adjust the bacterial suspension by diluting with saline until it matches the standard, re- shaking the standard before each comparison.

Total Viable Count (TVC) gives a quantitative idea about the presence of microorganisms such as bacteria, yeast and mold in a sample. To be specific, the count actually represents the number of colony forming units (cfu) per g ( or per ml) of the sample.

A surface viable count is made when the bacterium is best grown in surface culture or on an opaque medium. Ten fold dilutions of the bacterial suspension are made. A suitable volume of each dilution, e.g. 0.1ml, is pipetted on to the surface of each of the plates and at once spread widely with sterile glass spreader or sterile cotton swab. The plates are incubated at 37ºC for 24 hours. The viable count is calculated from the average colony count / plate. A viable count is essential to determine the level of the challenge inoculum.

Only plates (or replicate plates from the same dilution) with 30-300 colonies are counted. Plates with fewer than 30 colonies give statistically unreliable results, while plates with more than 300 colonies are too crowded to allow all the bacteria to form distinct colonies. Usually, more than one dilution in a series is plated, just to be sure that results in a countable range will be obtained. Ignore dilutions giving results outside of the countable range.

The concentration of bacteria in the original sample is calculated as:

 CFU ml-1 (or g-1) =

 colonies on plate

 final plate dilution

Frequently, volumes other than one ml are used to inoculate the plate. For example, 0.1 ml is often used when surface-plating, as larger volumes may not be absorbed by the agar.

For this reason, the size of the inoculum is usually incorporated with the dilution factor to give the "final plate dilution" (d x i). When 1.0 ml of a 10-4 dilution is plated, the final plate dilution is 10-4. When 0.1 ml of the same dilution is plated, the final plate dilution is 10-4 X 10-1 = 10-5.

The formula becomes:

 CFU ml-1 (or g-1) =

 colonies on plate

 d x i

Of the many media available, Muller Hinton agar is considered to be the best for routine susceptibility testing of nonfastidious bacteria for the following reasons.

It shows acceptable batch to batch reproducibility

It is low in sulphonamide, trimethoprim, and tetracycline inhibitors.

It gives satisfactory growth of most nonfastidious pathogens

Meat infusion, beef extract, acid hydrolysate of casein in the medium provide with C, H, N and other trace element sources.

Starch in the medium acts as a protective colloid against toxic substances.

Hydrolysis of starch during autoclaving provides with small amounts of dextrose which acts as a source of energy.

Nutrient Agar consists of peptone, beef extract and agar. This relatively simple formulation provides the nutrients necessary for the replication of a large number of microorganisms that are not excessively fastidious.

The beef extract contains water soluble substances including carbohydrates, vitamins, organic nitrogen compounds and salts.

Peptones are the principle sources of organic nitrogen, particularly amino acids and long chained peptides.

Agar is the solidifying agent.

Nutrient Agar is used for the cultivation of bacteria and for the enumeration of organisms.

Composition: Chlorexdine Gluconate Solution

I.P 1.5%v/v

Cetrimide Solution (strong)

Specified working range: 1:15 dilution.

Diluent: St. Distilled water.

Composition: Chloroxylenole IP 4.8 %, Terpineol BP 9.0

IP 13.1 % v/v (Alcohol)

Specified working range: 1:20 dilution.

Diluent: St. Distilled water.

Composition: Povidone-iodine

IP 5%w/v. (0.5%w/v available iodine).

Purified water IP.

Media: Muller Hinton Agar (200ml)

Nutrient Agar (40ml)

Diluent: Autoclaved distilled water (50 ml)

Sterile Saline (100ml).

Organism:

Glass ware: Sterile assay tubes (15)

(10 ml) Sterile glass pipette (5 pipettes)

Petri plates (15)

Miscellaneous: Auto pipette (vol. 1000ul, 100ul)

Sterile tips (vol. 1000ul, 100ul)

Cotton Swabs.

Instruments: Incubator at 37ºC.

Votex.

Autoclave

Oven

Laminar Air Flow Unit.

HiMedia Laboratories Pvt. Ltd.

Ingredients: gms/litre

Peptic digest of animal tissue 5.00

Beef extract 1.50

Yeast extract 1.50

Sodium Chloride 1.50

Agar 15.00

Direction of preparation:

Suspend 28.00 gms in 1000ml of distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15lbs pressure (121ºC) for 15 minutes.

The mouth of the nutrient agar flask was flamed to sterilize it and sufficient nutrient agar was poured into the sterile plate (approximately 20ml). The plate was then covered with the lid and then placed on a flat surface and slightly rotated it so that its agar was equally spread on the surface on the plate. The agar solidified in a span on 10 minutes.

2. Mueller-Hinton (MH) agar:

HiMedia Laboratories Pvt. Ltd.

Ingredients: gms/litre

Beef infusion from 300.00

Casein acid hydrolysate 17.50

Starch 1.50

Agar 17.00

Direction of preparation:

38.0 gms in 1000ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121ºC) for 15 minutes. Mix well before pouring.

The mouth of the Mullar Hinton flask was flamed in order to sterilize it and poured sufficiently into the plate (approximately 20ml). The plate was then covered with the lid and placed on a flat surface and slightly rotated it so that its agar was equally spread on the surface on the plate. The agar solidified in a span on 10 minutes.

3. Sterile saline:

To prepare saline water 100 ml of distilled water was poured into the flask and gms of sodium chloride was added to the distilled water. It was then autoclaved at 15 lbs pressure (121ºC) for 15 minutes.

4. Inoculum preparation:

Using colonies taken directly from an overnight (20 to 24 hou r) nutrient agar culture plate, a suspension of the test organism ( S. aureus ) was prepared in sterile saline. The suspension was adjusted to a turbidity equivalent to a 0.5 McFarland standard. This suspension contained approximately 1 to 4 x 108 CFU/ml. Within 15 minutes after adjusting the turbidity of the inoculum suspension, it was used for plate inoculation.

To check the growth of Staphylococcus aureus bacteria on the Nutrient Agar plates it is necessary to check the viable count of the bacteria. The 10 fold dilutions were carried out and 10-1, 10-2, 10-3, 10-4and 10-5 dilutions were plated.

Using colonies taken directly from an overnight (20 to 24 hour) nutrient agar culture plate, a suspension of the test organism ( S. aureus ) was prepared in sterile salinein a sterile assay tube. The suspension was adjusted to a turbidity equivalent to a 0.5 McFarland standard.

5 Sterile assay tubes were labeled as follows - 10-1, 10-2, 10-3, 10-4, 10-5

Using a sterile 10 ml glass pipette 4.5 ml of sterile saline was added to each tube.

0.5 ml of S. aureus suspension was transferred to the first assay tube labeled 10-1 using a 100 – 1000 ul autopipette .

The tube was vortexed and 0.5 ml of culture from this tube was transferred to the second tube labeled 10-2

Again this tube was vortexed and 0.5 ml of culture from this tube was transferred to the third tube labeled 10-3

Similarly dilutions were carried out for the remaining tubes.

After all the dilutions were done, 0.1 ml of the culture (using a 20 – 200 ul autopipette) from each tube was spread plated on respective nutrient agar plates using a sterile cotton swab.

All the plates were incubated at 37ºC for 24 hours.

Microbial Challenge test, which is established by internation practice, is a most widely used to method to evaluate the efficacy of antiseptics. The test is performed over a period of time to simulate clinical scenarios in the process of manufacturing and handling. In this test, samples are inoculated with the test microorganisms and then inspected visually and determine if the antiseptic system is effective. The microbial challenge test method is the classical means to evaluate antiseptic efficacy because it closely mimicks the practical situation

Varying dilutions of the antiseptic were made in sterile assay tubes.

To each dilution add 0.1ml of the culture ( using 20 – 200 ul autopipette ) was added from the inoculum prepared for viable count.

Each tube was vortexed and allowed to stand for 1 minute.

After 1 minute, 0.1ml ( using 20- 200 ul autopipette) from each tube was taken and spread plated on the respective Muller Hinton agar plates using sterile cotton swabs.

All the plates were incubated at 37ºC for 24 hours

1. Viable count:

Dilution

No. Of colonies

10-1

Mat growth

10-2

Mat growth

10-3

Too many to count (TMTC )

10-4

101 X 4= 404

10-5

12

2. Microbial challenge test:

Sample

Dilution

No. of Colonies

A

1:5

0

1:16

1

1:60

0

B

1:10

0

1:20

1

1:50

0

C

1:5

0

1:15

0

Full strength

0

10-4 = 101 x 4 = 404

= 404 x 104 x 10 = 4.04 x 107

~ 4 x 107 cfu/ ml.

10-5 = 12 x 105 x 10

= 12 x 106

= 1.2 x 107

~ 1 x 107 cfu/ ml.

Average: 4 cfu/ ml.

From the viable count the cell density of the initial inoculum was 4 cfu/ml.

Microbial challenge test:

For sample A

In the 1: 15 dilution and the 1: 60 dilution there is no colony growth. Whereas in the 1:16 dilution there is presence of 1 colony of Staphylococcus aureus Bacteria.

For sample B:

In the 1: 10 and 1:50 dilutions there is no colony growth. However in the 1: 20 dilution there is growth of a single colony of the bacteria.

For sample C:

In the dilutions of 1: 5, 1: 15 and full strength dilutions do not have any bacterial growth and therefore no colonies are observed on the plate.

From the observations it can be concluded that the sample A of the antiseptic is effective on staphylococcus bacteria within and beyond its range.

The conclusions that can be drawn from the observations are that the sample B of the antiseptic agent is effective within and beyond its range.

The sample C is effective on the staphylococcus bacteria this conclusion can be drawn from the observation of no bacterial growth on the dilutions of sample C.

Therefore from the results obtained it can be concluded that the staphylococcus bacteria is killed with the help of every antiseptic with varying concentrations in the specific constant time. This proves the effectiveness of the antiseptic. However it is recommended in hospitals and laboratories to change the antiseptic monthly because the bacteria become immune to the antiseptic after a certain period. Therefore with the help of this experiment I have checked the effectiveness of the antiseptics on the Staphylococcus aureus bacteria.

From the Microbial Challenge Test conducted it is observed that all the three antiseptics are effective with and beyond their respective range. However comparing the antiseptics among each other, it can be concluded that the sample C is most effective, since the Sample A and Sample B have 1 colony growth in one of the dilutions. However Sample C does not have any colony growth. The third sample is most effective because its composition includes Povidone- iodine which is not included in the other two samples. However a further question for research has been probed in this experiment to identify if these antiseptics cause any harm to the human body. There is a suspect that these antiseptics are toxic to the human body. Therefore these antiseptics are on account of further research.

The Mechanism of Action and Efficacy Evaluation Methods for Preservatives

Hongyu Xue, Guiqi Yin, Evelyn G. Su

Nanjing Zhongshi Chemical Co., Ltd.

http://www.merck.com/mmhe/sec17/ch188/ch188d.html

http://agrc.ucsf.edu/supplements/immune/immune_system_06.html

http://www.bd.com

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The Effect Of Different Concentrations Of Enzyme Biology Essay

In this experiment, I am going to determine the effect of different concentration of enzyme catalase on the rate of reaction of decomposition of hydrogen peroxide. Normally, hydrogen peroxide is produced naturally in human or plant cell. Hydrogen peroxide is the by-product of respiration. As an oxidizer, it will decompose to form oxygen and water. The chemical equation for the decomposition of hydrogen peroxide is 2 H2O2 ? 2 H2O + O2. The reaction is speeded up by the presence of enzyme, namely catalase which is used in this experiment. This mechanism is important in living organisms’ cells and body system particularly in human. This is because the corrosive characteristic of hydrogen peroxide may damage the wall of liver where it is largely produced during cellular respiration process. When it is present in high concentration, it is an aggressive and powerful oxidizer, whereby it is unstable and also hazardous as it will corrode many substances including human skin. Therefore, concentration of hydrogen peroxide in the cell should be constantly regulated. When hydrogen peroxide is used for the purpose of experiment, this highly corrosive material should be kept in a container made up of non-reactive material such as glass. However, at low concentration, hydrogen peroxide can be used as disinfectant and antiseptic for medicinal uses.  

C:\Users\User\Documents\yap in\bio report\enxyme\Hydrogen_peroxide_files\120px-Wasserstoffperoxid.png C:\Users\User\Documents\yap in\bio report\enxyme\Hydrogen_peroxide_files\120px-Hydrogen-peroxide-3D-balls.png

Diagram 1 : Chemical structure of hydrogen proxide, H2O2

In this context, catalase, a teramer of four polypeptide chains is made up of over 500 amino acids long. It is also categorised as globular protein in which the polypeptide chain is highly folded into a compact spherical shape. There is also active site available to bind to the hydrogen peroxide substrate to form enzyme-substrate complex.  It is further adapted with four porphyrin heme groups to react with hydrogen peroxide. Besides, the enzyme catalase is known to be one of the enzymes that possess a high turnover number. Its turnover number can be up to 600 000 whereby one molecule of enzyme catalase can catalyse the decomposition of 600 000 molecules of hydrogen peroxide to oxygen and water at body temperature.  This reaction is known as catabolic reaction as the hydrogen peroxide molecule is broken down into oxygen and water which are comparatively smaller. Sometimes, catalase also uses hydrogen peroxide to oxidise toxins including Phenols, Formic Acid, Formaldehyde and Alcohols. In this experiment, potato is chosen to be tested due to the presence of catalase in it. However, other organisms such as fungi or yeast can be used as well as they are producers of enzyme catalase.

C:\Users\User\Documents\yap in\bio report\enxyme\Catalase_files\250px-Catalase_Structure.png

Diagram 2 : The structure of catalase

Enzyme is used to speed up the rate of reaction by lowering the activation energy of a reaction. Activation energy or free energy of activation, is the initial investment of energy for starting a reaction – the energy required to contort the reactant molecules so the bond can break for a reaction to occur.  Enzyme functions as biological catalyst in many chemical reactions that occur inside our body. For example, saliva secretes enzyme amylase which catalyses the hydrolysis of carbohydrates in the mouth.  Not only does enzyme play an important role in maintaining efficient function of body system, it is largely used in industrial field as well to speed up the production rate. For example, protease is commonly used in biological detergent for domestic washing and rennin is used in manufacture of cheese. For an enzyme to carry out its function effectively, active site should present on the surface of the polypeptide chain. An active site is a groove or pocket formed by the folding pattern of the protein. This active site has particular chemical composition and electrical charges on the amino acids, which make up the specificity of the enzyme, in which it allows only certain substances to bind to it. When the substrates bind to the active site, here the working mechanism of enzyme starts. The binding of the substrate to the active site bring the substrates closer and thus aids in bond formation in anabolic reaction. In catabolic reaction, the active site may distort the shape of substrate to break its bond. When the products are formed, the substances no longer fit into the specific shape of the enzyme and will leave the active site of the enzyme. The enzyme is free to bind to another substrate and catalyse another reaction. The enzyme is not altered at the end of reaction.

C:\Users\User\Documents\yap in\bio report\enxyme\Enzyme_files\450px-Induced_fit_diagram.png

Diagram 3 : The working mechanism of enzyme

As enzyme contains specific shape and charge on its active site, its activity is easily affected by the changes in the surrounding conditions. Generally, different pH, temperature, concentration of substrate or concentration of enzyme has a large impact on its efficiency in carrying out its function. Whenever the changes in surrounding such as change in pH or temperature alter the bonding between the R group of the amino acids in the polypeptide chain which form the active site, the shape of active site will change and thus the substrate will no longer bind to the site. At this point, the enzyme is said to be denatured. On the other side, when the temperature or pH is optimum for the reaction, the rate of reaction is the highest. Although the optimum pH and temperature may vary from one another, optimum temperature for most enzymes functioning in human body system is often 37 ºC. However, the presence of inhibitors or cofactors may alter the enzyme activity as well. In this experiment, the effect of enzyme concentration is chosen to be investigated on the rate of reaction catalysed by enzyme catalase. An increase in enzyme concentration will increase the active site available and thus increase the rate of reaction until it reaches maximum velocity when all active sites of the enzyme molecules are engaged.

Do different concentrations of enzyme affect the rate of reaction?

To investigate the effect of different concentrations of catalase on the rate of reaction to catalyse the decomposition reaction of hydrogen peroxide

To determine the presence of catalase on the rate of reaction of hydrogen peroxide.

To develop effective experimental skills throughout the experiment

To determine the effect of different concentrations of enzyme on the enzyme activity

The higher the concentration of enzyme, the higher the rate of reaction until a maximum velocity is reached.

Use a water displacement technique to determine the volume of oxygen gas evolved

Calculate the rate of reaction by using the gradient of the graph

Freshly mashed or blended potato, 3.0 % hydrogen peroxide solution, buffer solution (pH 6.5), distilled water

Boiling tubes, graduated tubes, 500 ml beaker, weighing balance, spatula, delivery tube, stop watch, measuring cylinder, dropper, rubber bung, weighing dish

Variable

How the variable is determined

1. Manipulated

Concentration of catalase

By using different mass of blended potato at 1g, 2g, 3g and 4g. Different masses of blended potato indicate the difference in concentration of catalase in its content.

2. Responding

The volume of oxygen gas released

By recording down the reading on the graduated tubes at 30 seconds interval.

3. Constant

pH

Volume and Concentration of hydrogen peroxide

By using buffer solutions at pH 6.8 throughout the experiments

By using the same volume and concentration of hydrogen peroxide, which is 2.5cm3 of 3.0 % hydrogen peroxide throughout the experiment

1 g of the freshly prepared or blended potato is transferred into a boiling tube.

5 cm3 of buffer solution is added into the tube and it is swirled to mix the substrate.

A graduated tube is filled with water to the brim.

It is placed carefully into a beaker of water. One end of the delivery tube is placed into the graduated tube with the other end with rubber bung ready to fix with boiling tube.

2.5 cm3 of hydrogen peroxide solution is measured and it is added into the boiling tube containing the potato and buffer solution.

The tube is immediately closed with a rubber bung connected to the delivery tube. A stopwatch is started by one member of the pairs in conducting this experiment.

The volume of gas released is measured for every 30 seconds for 5 minutes or until the gas evolution stops.

The experiment is repeated using 2g, 3g and 4g of freshly blended potato.

The results obtained are recorded in a table.

Graphs for volume of gas released against time is plotted for each concentration or amount of enzyme used.

The initial rate of reaction for each concentrations of enzyme used are worked out.

Time /s

Volume of oxygen gas collected at potato with mass of

1 g

2 g

3 g

30

0.8

8.6

12.8

60

3.5

14.0

19.0

90

4.8

18.0

22.2

120

6.0

21.2

24.8

150

6.8

23.4

30.8

180

7.6

25.0

38.6

210

8.2

27.0

48.0

240

8.4

29.8

58.8

270

8.5

33.2

70.8

300

8.7

34.4

83.6

Table 1: The volume of oxygen gas collected at 30 intervals with different mass of potato.

Formula 1 : The mean value of the date obtained

Formula 2 : The initial rate of reaction

1g of blended potato

78-24 = 54

4.5 -1.2

= 3.3

Expected line

2 g of blended potato

17 – 6

=11

3g of blended potato

4g of blended potato

1

0.0611

2

0.2895

3

0.6579

4

0.7000

Based on the above experiment, the effect of different concentrations of enzyme on the rate of reaction is successfully determined. Five graphs are plotted based on the results obtained in the experiment to show the data in a clearer way and provides a better mean for analysing. The results show that the rate of reaction is increased by an increase in enzyme concentration. In this experiment, potato is used as source of catalyse. The first four graphs showing oxygen gas evolved against time are drawn based on respective mass of blended potato used. The initial rate of reaction is measured from each graph by obtaining the gradient of the graph. A predicted line is drawn on each graph. Generally, the longer the time taken, the higher the volume of oxygen gas evolved. In the beginning, all graphs show an rapid increase , the speed is the slow down as some of the substrates are converted to products. For the substrate at 1 and 2 g of bended potato used, the maximum volume of oxygen gas evolved has reached within 300 seconds and a plateau is obtained. This is because the reaction has completed for all substrates. Theoretically, the maximum volume of oxygen gas released should takes a shorter time as compared to 1g and 2 g of potato as more active site are offered. However, In the 3 and 4 g of blended potato which react, the maximum volume of oxygen is unable to be obtained within 300 seconds. This is probably due to some errors conducted throughout the experiment, particularly due to the vigorous and rapid reaction and in the process of changing the graduated tube. The errors will be discussed later. The initial rate is taken because the rate of reaction is rapid as the collision between the substrate and enzyme is the highest. The rate of reaction may not be reliable to be compared between data if readings are taken in the middle of the experiment because some reactions have reached the maximum rate. The initial rate of reaction for hydrogen peroxide with 1g, 2g, 3g and 4g of blended potatoes are 0.0611, 0.2895, 0.6579 and 0.7000 cm3/ s respectively.

The initial rate of reactions for all the experiments are then compiled into the fifth graph. This shows a clearer picture on the effect of concentration of substrate on the rate of reaction. Initially, there is an increase in the rate of reaction when the mass of blended potato increases. This is because the increase in the concentration of enzyme offers more active site for the binding of substrate. Then, the slope of increasing line becomes less steep with further increase in concentration of enzyme. This is because the active site has been occupied by the substrates or it is said to be saturated whereby the increase in substrate has no further effect on the rate of reaction. Theoretically, the graph should reach a maximum velocity where the plateau occurs in the graph. However, in this experiment, the plateau is not shown because most probably the concentration of enzyme is not high enough to bind to all the 3.0 % of hydrogen peroxide substrate.

However, throughout the experiment some errors might occur in which the real values may not be obtained. Firstly, there is a high tendency for the reading obtained from water displacement method to be inaccurate especially when the volume of oxygen gas evolved are too much that the first graduated tube is fully filled with oxygen gas and when the delivery tube has to be transferred to the next prior-prepared graduated tube. The delivery tube transferring process may consume some time particularly if a rubber delivery tube is used instead of a glass delivery tube. This will cause some of the oxygen gas to escape into the water during the process. Next, parallax error may occur as well when the reading is taken from the graduated tube on the volume of oxygen gas evolved. This is because oxygen gas is a colourless gas, in which its level is not so clearly seen on the calibration of the graduated tube. To minimise the errors, the experiment is repeated twice and the mean reading is obtained. To further increase the accuracy of the results, a piece of white paper can be placed behind the graduated tube to make the reading easier. Next, the possible error is greater if the experiment is carried out individually. This is due to the human limited ability to record the reading and at the same time watch over the time. Inaccuracy may arise. In this case, a pair work is preferred in this experiment as one of the members times and the other one record the readings obtained. Next, when the mashed potato is poured into the boiling tube from the weighing dish, some potato may be left in the weighing dish. To minimise this error, a few drops of distilled water can be used to rinse the weighing dish to ensure there is no residue left.

Consequently, there are a few precautions that ought to be taken to increase the accuracy of the results obtained. For each experiment, the potato used must be freshly mashed or blended. If the potato is prepared in a container, the lid of the container should be kept closed after the desired mass of blended potato is scooped out for each and every experiment. The preparation of blended potato in a beaker which is exposed to the air should be prevented because oxidation will occur and this may affect the activity of enzyme catalase in it. Changes in surrounding such as temperature may also induce changes in the enzyme. A blended potato is used instead of discs of potato so that it will react easier. Its viscosity should be reduced so that it is easier to use. Next, hydrogen peroxide has to be stored in an opaque container as it breaks down quickly when exposed to light. The lid of the container that contains hydrogen peroxide solution should be kept closed after each desired sample is taken out using a dropper as the oxygen in the surrounding air may oxidise its content and causes the results to be inaccurate. A buffer solution is used to ensure the pH is kept constant throughout the experiment. The buffer solution of citric acid sodium phosphate solution which has a pH of 6.8 is used because this is the optimum pH for the enzyme catalase. Furthermore, a water bath is preferable as the surrounding temperature may change throughout the experiment. In addition, as the rubber bung of the delivery tube should be of the same size as the boiling tube to ensure all the opening of the boiling tube containing enzyme and substrate is fit tightly, it should be pushed and twisted with care. It should also be checked from time to time to ensure there is no leakage of product in gaseous form to the surrounding. Besides, the other open end of delivery tube should be placed in water all the time for the bubble of gas to form and rise to its surface. The presence of air bubbles ensure that the rubber bung is still in contact with the boiling tube unless the substrate and enzyme has completely reacted. To fix the graduated tube in place, a retort stand and clamp can be used. Besides, the boiling tube containing reactants and enzyme ought to be swirled throughout the experiment to ensure the substrate and enzyme react. This may increase the rate of collision between the reactants and enzymes and thus fasten the time taken for the reaction to complete.

Throughout the experiment, some safety measures should be abided by. As the substrate used in this experiment which is hydrogen peroxide is highly corrosive, rubber glove should be used to protect the skin. After the hydrogen peroxide is used, it should be disposed off and not to be returned to stock bottles as any contaminants may result in decomposition and explosion may occur. The blended potatoes have to be handled carefully as well as it will irritate some people’s skin. A lab coat should be put on. The glass wares and the delivery tube used should be handled carefully as they are fragile.

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The hypothesis is accepted. The presence of enzyme increases the rate of reaction of hydrogen peroxide. When the concentration of enzyme increases, the rate of reaction increases until a maximum velocity is reached.

The species of potato

Different species of potato may contain various concentration sof catalase

The age of potato

An older potato may have lower concentration of catalase

The freshness of potato

The concentration of catalase may vary in different potatoes which are stored in different ways before experiment. Storage at high temperature may cause the enzyme to denature

Part of potato used

Different parts on the potato may have different amount of catalase.

The effect of temperature on the enzyme activity

The effect of different concentrations of substrate on the enzyme activity

The effect of pH on the enzyme activity

The effect of concentrations of enzyme on activity of other type of enzyme such as amylase on starch

The effect on the rate of reaction of hydrogen peroxide by using different concentration of fungi as the source of catalase

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