Investigation of the Probiotic Properties of Bacterial Strains from Two Probiotic Drinks and Their Survivability in Artificial Gastric Juice

Investigation of the probiotic properties of bacterial strains from two probiotic drinks and their survivability in artificial gastric juice ABSTRACT: Two probiotic drinks were investigated in vitro to test their ability to survive acidic conditions and their probiotic factors. Both the products: Actimel and Yakult contain gram-positive bacteria, but Actimel also has a gram-negative bacteria. The ability to survive was investigated by adding artificial gastric juice to the products and incubating at different times.Actimel and Yakult were both able to survive the gastric juice. Actimel produced more colonies than Yakult but they both lost the same percentage of viability. The longer the time incubated the more the loss of viability. Introduction: In recent years health promoting functional foods has entered the global market as a result of increased prevalence of lifestyle related diseases (A. A. Aramide et al, 2009). People use functional foods and diet to maintain optimal health. C onsumption of probiotics is one of the ways someone could reach and maintain their optimal health.A probiotic is â€Å"living microorganisms, which upon ingestion in certain numbers, exert health benefits beyond inherent basic nutrition† (Todd R. Klaenhammer, 2000). According to the WHO/FAO report 2001 these probiotics can help prevent disorders associated with the gastrointestinal tract, diarrhoea caused by certain pathogenic bacteria and viruses, inflammatory diseases, allergies and a lot more. Actimel and Yakult is a couple of the said probiotic drinks. They claim to increase your body’s natural defences by fighting off the â€Å"bad† bacteria. Actimel is a yogurt-type drink produced by a company called Danoneâ„ ¢.It has three strains of bacteria, two traditional yoghurt cultures: Lactobacillus bulgaricus  and  Streptococcus thermophiles and a third one called L. casei Imunitass ® (http://www. actimel. co. uk/About/-WhatIsActimel. aspx, Accessed Feb, 28, 2010). Lactobacillus is a genus of bacteria that aid in the conversion of lactose to lactic acid hence increasing acidity in the stomach making it hard for harmful bacteria to survive (http://en. wikipedia. org/wiki/Lactobacillus, Accessed Feb 28, 2010). Actimel contains 10 billion L. casei Imunitass ® bacteria per 100ml bottle.This bacterial strain works under a wide range of pH and temperature hence able to survive the acidic conditions in the stomach. This ensures that the bacteria reach the gut alive and active. It helps by topping up the good bacteria in the stomach and making it hard for the germs to survive. The bacteria also aids in strengthening the gut wall so that only certain nutrients can pass. In 2004 a trial carried out to find the effect of Actimel on the immune response of subjects under academic examination stress showed that Actimel was able to control the number of lymphocytes and CD56 cells in subjects under academic examination stress.Other studies also show that the Actimel bacterial strains can be used in treating allergic rhinitis, prevention of diarrhoea and induce immune responses. On the other hand Yakult is milk based probiotic and contains only one strain of bacteria: Lactobacillus  casei  Shirota. It is produced and distributed by Yakult Honsha Co. Ltd. It contains 6. 5 billion L. casei Shirota per 65ml bottle. A variety of scientific studies have shown that Yakult has an effect on the human NK-cell activity, intestinal micro flora and immune parameters in humans.As a guideline a probiotic microorganisms should be resistant to gastric juices and be able to grow in the presence of bile under conditions in the intestines. The aim of this experiment is to measure the survivability of the strains in artificial gastric juice and to identify the bacterial strains said to be in the product. MATERIALS AND METHODS: Gram Stain: Firstly the bacteria were heat fixed according to the instruction in the lab manual. After heat fixing , crystal violet stain was added to the bacteria for 2 minutes, then washed in water and Lugol’s iodine for 30 seconds.The bacteria were decolorised by adding 95% alcohol for 15 seconds followed by a water wash and counter stain with safranin for 1 minute. This was then washed with water and examined under high power (x100) using oil immersion. A picture of these strains each from Actimel and Yakult directly and pure culture was taken. DNA Extraction: To extract the DNA, 1 ml of culture was centrifuged for 5 minutes. The pellet was re suspended in 480 ? l of 50mM EDTA with 60 ? l of 10mg/ml lysozyme then incubated at 370C for 45 minutes, centrifuged for 2 minutes and re suspended in 600 ? of nuclei lysis solution and incubated at 800C for 5 minutes. After cooling down 3 ? l of RNAase was added and left to incubate at 370C for 30 minutes. The mixture was left to cool and 200 ? l of protein precipitation solution was added, left on ice for 5minutes followed by high speed (13000 rpm) centrifuging for 5 minutes. The supernatant was then added to 600 ? l of isopropanol and mixed until DNA â€Å"threads† were formed and centrifuged for 15 minutes. The DNA pellet was washed with 200 ? l of 70% ethanol and centrifuged for 2 minutes. The ethanol was then removed and the DNA left to air dry and then re suspended in 50 ? of sterile water. PCR of chromosomal DNA: A 2 ? l of the DNA was added to 1 ? l of AmpF primer(GAGAGTTTGATYCTGGCTCAG), 1 ? l of AmpR (AAGGAGGTGATCCARCCGCA) primer, 2 ? l of dNTP’s, 10 ? l of x10 PCR buffer, 83 ? l of water and 1 ? l of Taq polymerase was added. This mixture was placed in the Promega Wizard Chromosomal DNA preparation kitâ„ ¢ and run according to the manufacturer’s guidelines. PCR Purification: The PCR reaction contents were added to a 1. 5 ml Eppendorf tube with 500 ? l of buffer PB1. This was centrifuged at high speed in the spin column for 30 seconds.A 750 ? l of buffer PE was added to the spin column and centrifuged for 1 minute. The spin column was then placed in an Eppendorf tube and 50 ? l of water was added and centrifuged for a further 1 minute. A 15 ? l of this PCR product was added to 5 ? l of Gel loading buffer and was run at 50 V for 2 hours. 20 ? l of the PCR product was then sent to the John Innes sequencing service for sequencing. Media Preparation: To media was prepared by adding 37g of Brain Heart Infusion (BHI) to 1 litre of distilled water and mixed using a magnetic stirrer.This was then added to a conical flask with 3g of agar and autoclaved at 1210C, 15 psi for 10 minutes. The media was then microwaved and poured onto petri dishes with Bunsen burner going, to sterilise the air around. Survival Studies: For carrying out the survival studies, 5 ml of the product was added to 25 ml of artificial gastric juice and left to incubate at 370C for 30, 60 and 90 minutes. The product was taken from different bottles to ensure replicates. After incubation the mixture was then diluted to 10-5 for Yakult and 10-7 for Actimel. This was spread onto a petri dish and was left to incubate.The plates were then counted and the number of CFU/ ml was calculated. RESULTS: Culturing bacteria: Firstly the number of colony forming unit (cfu) per ml was worked out by culturing the bacteria from the probiotic products and counting the number of colonies formed. This was then used to work out cfu/dose by using the volume they are produced in, which are 100 ml and 65 ml of Actimel and Yakult respectively. Table 1: Class data of cfu/ml and cfu/dose of bacteria in the product Yakult(cfu/ml)| Yakult(cfu/dose)| Actimel(cfu/ml)| Actimel(cfu/dose)| 4. 21. x 109| 2. x 1011| 4. 36 x 109| 4. 36 x 1011| 4. 14 x 109| 2. 86 x 1011| 2. 6 x 108| 2. 6 x 1010| 9. 7 x 10 9| 7. 8x 1010| 2. 1 x 109| 2. 1 x 1011| 1 x 109| 6. 3 x 109| 7. 5 x 108| 7. 5 x 1010| 1. 6 x 109| 6. 5 x 1010| 5. 5. 2x 108| 5. 5 x 1010| 9 x 107| 5. 8 x 109| 1 x 1010| 1 x 1012| 7 x 107| 4. 5 x 109| 2. 5 x 109| 2. 5 x 101 1| 4. 6 x 109| 2. 99 x 1011| 1. 21x 109| 1. 21x 1011| 1. 68 x 108| 1. 09 x 1010| 4. 3 x 1010| 4. 3 x 1012| 4. 02 x 108| 2. 61 x 1010| 1. 18 x 109| 1. 18 x 1011| 9. 1 x 107| 5. 9 x 109| 2. 89 x 108| 2. 89 x 1010| 1 x 108| 6. 5 x 109| 2. 7 x 109| 2. 7 x 1011| x 108| 3. 2 x 1010| 3. 6 x 109| 3. 6 x 1011| 3. 4 x 107| 2. 2. x109| 2. 7 x 109| 2. 7 x 1011| 2. 39 x108| 1. 5 x 1010| 3. 78 x 109| 3. 78 x 1011| 9. 7 x 107| 6. 3 x 109| 5. 0 x 1010| 5. 0 x 1012| 1 x 108| 6. 5 x 109| 1. 4 x 109| 1. 4 x 1011| 1 x 108| 6. 5 x 109| 2. 6 x 109| 2. 6 x 1011| To compare the mean differences between these two products an independent t test was carried out assuming equal variance. Table 2: Independent t-test of the class data for cfu/dose on Actimel and Yakult Independent t-test| | | Mean| Standard Deviation| SE Mean| P Value| cfu/dose| Actimel| 7. 9 x 1011| 1. 45 x 1012| 3. 41 x 1011| 0. 056| | Yakult| 6. 29 x 1010| 1. 04 x 1011| 2. 46 x 1010| | The mean shows that Actimel contains 10 times more bacteri a than Yakult on average. But only the mean is not significant to come to a conclusion as this could be because of sample variation. The P value from the t-test is 0. 056 which is greater than 0. 05 (P>0. 05) hence the difference between the mean of the two products are not significantly different from zero at the 5% confidence level. Gram Stain: Figure 1 shows the gram stain images from Actimel (i) and Yakult (ii).Figure 1 shows the gram stain images from Actimel (i) and Yakult (ii). Gram stained slides of both Actimel and Yakult were captured onto a computer at x1000 magnification. From the images you can see that Yakult is stained all in one colour but the Actimel contains two different coloured stains. Survival studies: To test the survivability of the bacteria they were incubated with artificial gastric juice for 30 60 and 90 minutes. The colonies were then counted Table 3: Viable counts of survival studies at different time and different replicates | Actimel|Time/min| 1| 2| 3| Mean| CFU/ml| CFU/dose| 0| 329| 69| 1088| 371. 5| 3. 72 x 1010| 3. 72 x 1012| 30| 321| 39| 880| 322. 5| 3. 23 x 1010| 3. 23 x 1012| 60| 309| 28| 740| 286. 8| 2. 87 x 1010| 2. 87 x 1012| 90| 204| 24| 642| 238. 8| 2. 39 x 1010| 2. 39 x 1012| | Yakult| | 1| 2| 3| Mean| CFU/ml| CFU/dose| 0| 312| 135| 53| 125. 0| 1. 25 x 108| 8. 13 x 109| 30| 190| 134| 11| 96. 3| 9. 63 x 107| 6. 26 x 109| 60| 159| 130| 11| 92. 5| 9. 25 x 107| 6. 01 x 109| 90| 149| 84| 8| 81. 5| 8. 15 x 107| 5. 3 x 109| The table shows that colonies on both Actimel and Yakult decrease over time in all the replicates.Both the products decreased to about 65% of its original count. A graph (Figure 2) was plotted with the CFU/dose against time on a log scale and it showed a linear decline over time in both the products. DNA Extraction: Figure 3 shows the Chromosomal DNA gel image. Figure 3 shows the Chromosomal DNA gel image. The DNA from the bacteria was extracted and gel electrophoresis was carried out to ensure that a DNA was obtained from the extraction procedure. Lanes 3 and 4 have migrated towards the positive side showing that chromosomal DNA was obtained.PCR Purification: After the DNA underwent the PCR process, the PCR product was purified and run on a gel electrophoresis to check if PCR product has been obtained. Figure 4 shows the image of PCR product run under electrophoresis. Figure 4 shows the image of PCR product run under electrophoresis. As the image shows there is a PCR product obtained as there is a distinct band in lanes 2 and 3. DNA Sequencing: The PCR product was then sent to the John Innes centre for sequencing and the following sequence was obtained.Actimel: GGGTCGGGGCGGGTGCTATACATGCAGTCGAACGAGTTCTCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCC AAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAA Yakult: TAGGAGTGGGCGCGTGCCTATACATGCAAGTCGAACGAGTTCTCGTTGATGATCGGTGCTTGCACCGAGATTCAACATGGAACGAGTGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCCTTAAGTGGGGGATAACATTTGGAAACAGATGCTAATACCGCATAGATCCAAGAACCGCATGGTTCTTGGCTGAAAGATGGCGTAAGCTATCGCTTTTGGATGGACCCGCGGCGTATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCGATGATACGTAGCCGAACTGAGAGGTTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTC CACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGCTTTCGGGTCGTAAAACTCTGTTGTTGGAGAAGAATGGTCGGCAGAGTAACTGTTGTCGGCGTGACGGTATCCAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGTTTTTTAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAAGCGCATCGGAAACTGGGAAACTTGAGTGCAGAAGAGGACAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTGTCTGGTCTGTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCCTGGTAGTCCATGCCGTAAACGATGAATGCTAGGTGTTGGAGGGTTTCCGCCCTTCAGTGCCGCAGCTAACGCATTAAGCATTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACGGGGCCCGCACAAGCGGTGGGA Figure 5 shows the graphical summary of â€Å"strong† hits in the database of Yakult (i) and Actimel (i). Figure 5 shows the graphical summary of â€Å"strong† hits in the database of Yakult (i) and Actimel (i).This sequence was then run through the BLAST analysis to identify the probiotic isolate. Discussion: A Probiotic must be able to survive the conditions of the stomach and pass through to the gu t without significant loss. The bacteria found in the probiotics are cultured on petri dishes to test the amount of colonies present in the product. As mentioned above Actimel contains 10 billion per 100 ml and Yakult contains 6. 5 billion per 65 ml. From the t-test there was no significant difference in the content of the two products (Table 1). This was due to the fact that they both contain 100 million bacteria per ml of product. From the gram stain images both Actimel and Yakult was stained with the same conditions.But Yakult had only one stain whereas Actimel had two different stains. This is due to the fact that there is more than one species of bacteria in Actimel. The colour of the staining represents two different types of bacteria: gram-negative and gram-positive. All species of the lactobacillus genus are gram-positive. Gram-positive organisms retain the stain when they are stained with crystal violet but gram negative organisms lose their purple/violet stain when washed with alcohol but when retain safranin stain. Therefore the Yakult contains only gram positive bacteria (L. casei Shirota ®) while Actimel contains both gram positive and gram negative bacterium (Figure 1). From the survival studies we can

The Perceptions of American Women about “New Beauty Therapy Services for Kids”

The issue of beauty therapy among American women and sometimes men has been around for a long period that no one can really determine, however, the society has undergone great civilization/modernization and recently beauty salons for young kids have started emerging. These salons offer all sort of beauty therapy services ranging from manicure, pedicure, facials and many other beauty therapy services to young girls, due to the fact that the idea has not been in the market for along time the few salons that offer beauty therapy services to young girls charge a lot of money. Nevertheless, this new trend has received both positive and negative sentiments from the American public. I recently carried out a study to investigate the perceptions of the people towards this new idea. I developed a short questionnaire consisting of five questions and distributed them to ten literate and grown up women with young daughters between the ages of three and eight, within my neighborhood, Brooklyn. The questionnaire comprised of questions that were sensitive to various respondents’ perceptions as they allowed for the choosing of more precise answers. [Russ-Eft, D. F. 1980)] For instance, the second question required them to state whether they supported the idea of kids’ beauty therapy services or not, with answer options ranging from, â€Å"I strongly support, I support, I somewhat support, I strongly oppose, I oppose, I somewhat oppose. † The other three questions were depended on the answer to the first question and the second questions. The quest ions were dispatched through a reliable delivery method (hand delivery) and enough time provided for the answering of the questions, the respondents were also advised not to seek assistance from other people. As expected the survey yielded varying responses, with 80% of the respondents indicating that they are aware that kids beauty therapy services have been introduced in the market, while the rest indicated that they are not aware of the new service. Those who were not aware of the new kids’ beauty therapy services were discontinued from the interview as the answers to the rest of the questions depended on the knowledge of the new kids’ beauty therapy service. Interestingly only a paltry 20% of the survey sample who knew about the new kids beauty therapy services indicated that they â€Å"strongly supported† the new service and a further 20% indicated that they â€Å"somehow supported the new service. † 40% indicated that they â€Å"strongly opposed the new service† and the remaining 20% showed that they â€Å"opposed the new service to kids. † Since the answering of the other three questions of the study was dependent on the answer to question number two only 40% of the respondents went on to answer the remaining questions. This is so because the other three questions were meant to elicit the answers as to what needed to be done and what should not be done about the new beauty therapy service to the kids. It was therefore irrelevant for respondents who did not support the idea to continue answering the other questions as they were bound to give out unreliable answers since in the first place they did not have any interests on the new service. [Wentland, E, J. & Smith, K. W. (1993)] Out of the 40% of the survey sample that proceeded with the rest of questions (by virtue of their support to the new kids’ beauty therapy service) 20% indicated they have once or twice taken their young daughters to the kids beauty therapy salons while the remaining 20% showed they have never done so but they were planning to do so in future. Interestingly 30% agreed that indeed the services are good for their young daughters but they are being overcharged and therefore the charges need to be adjusted. The remaining 10% indicated that the charges were reasonable compared to the good beatification services done to the young kids. On the question of whether some services currently in the kids’ beauty therapy package should be scrapped, they all (100%) agreed that some services needs to be removed from the package as they just did not make sense to young kids. [Wentland, E, J. & Smith, K. W. (1993)] The overall response of the five questions was very reliable as it systematically and precisely gave out information on the perceptions of the respondents. From the results this is visible from the answers to question one through question five. The questions were also arranged in a logical manner to avoid clue giving, those who gave â€Å"NO† as their answer to question one were discontinued from the interview as the study was dependent on the knowledge of the issue being investigated i. e. new beauty therapy services for kids. Further, those who had their answer as â€Å"I strongly oppose/ I oppose/I somewhat oppose† for question two were similarly discontinued from the interview. The remaining questions of the survey were about what needed to be done or not about the new service and therefore it was in order to discontinue those who did not know about the service or support it. The main reason behind this was to avoid false and unreliable answers as those did not support the service did not have any business to comment as to what needs to be done or not about the new service. [Russ-Eft, D. F. (1980)] The simple survey comprising of five-question questionnaire gave out very precise information that could have otherwise not been possible if heavily worded questions were used. This helped the respondents to perceive the questions as not bothersome or requiring much of their time and energy and therefore they gave out correct answers according to their perceptions (or lack of them) on the issue being investigated. Again, the survey sample was small (ten literate women) and the questionnaire comprised of simple questions with instructions written in bold attached on core questions to help extract valid and reliable data. The language used in the questionnaire was simple and unambiguous, further still, the questions were very sensitive in order to extract finer details from the respondents, for instance question number two was very prompting to the respondents as it gave six options for answer. Russ-Eft, D. F. (1980)] In conclusion the questionnaire met all the requirements of the specific criteria of a good measurement i. e. reliability, validity, and sensitivity. It is reliable because that gave out results that could repeatedly be got if the same sample was to be used again; it was valid because it followed a systematic procedure and gave out valid results, and it was sensitive because it allowed respondents a more options for answers. [Russ-Eft, D. F. (1980)]

Musical cultures of native american and brazil Essay

Musical cultures of native american and brazil - Essay Example "Traditional music culture in Brazil and Native American cultures" essay describes the diversity of music styles of these cultures. These two groups of people have had a long history characterized by struggle, the strife and the final triumph. To begin with, The American Song published by Alexander Street Press. It is a large database that contains over 50,000 tracks which offer room for people to listen to and have a feel of America's past music. As such the database includes songs formulated by the Native Americans, the immigrants as well as slaves. Also inclusive in the database are Civil Rights songs, the political campaigns, Civil War among much more. The Encyclopedia of Natives Music: More Than a Century of Recording from Wax Cylinder to the Internet by Brian Wright-McLeod, illustrated by photographs and cover albums. And printed by Tucson: The University of Arizona Press, 2005 for Fine Art Music Collections provides vital information concerning the Native American culture. As such, the Encyclopedia of Native American Music recognizes contributions made by some Native recording artists by examining commercially released music history. Indeed it provides an overview of the recorded Native music while pointing out its historical value which has been organized by the genre for a much quicker reference. In addition, soundtracks as well as, compilation albums for artists have been included. As such this book enumerates some spoken word recordings and further includes comedy, poetry and audio books. Indian Blues: American Indian and Politics of Music, 1879-1934 written by, John W.T,    Norman: University of Oklahoma Press, c2009. As such, Troutman  examines the politics of music on the Indian reservations and public venues as well as, reservation boarding schools at the beginning of the 20th century. During this period , US government (Office of the Indian Affairs) was engaged in controlling the musical practices Indian Americans as a way of assimilating them. The author uses the opening of the Carlisle Indians School in 1879, and the enactment of the Indians Reorganization Act in 1934 at the start and conclusions. As such, he examines how the Native American, government officials as well as , the non-Indian audiences adopted musical practice in order to shape the Indian policy. Music of the First Nations: Traditions and Innovations in the Native North American edited by Tara Browner.  Urbana: University of Illinois Press, c2009.  As such, this anthology lays out a numb er of ways in approaching an ethnomusicology of the musical expression of the Native Americans. Concerning the Culture in Brazil, About.com Latin music describes different types of traditional songs performed in Brazil, more particularly; the samba, Bossa nova and Choro have been identified. Additionally, Buzzlle describes different forms of Brazilian music, and the musical instruments used, listing, Bateria, Ganza Shekere, and the tambourine among others RESPONSE TO III The challenges include language diversity, rise of global communications and the ease at which people move, increased competition, the American cultural exchange service, the Bahia

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A Segment from the film Finding Nemo - Essay Example Finding Nemo is one of the most successful animation film blockbusters. It was released in 2003 worldwide and took the entire world by surprise. Its stunning animation, the astounding undersea sceneries, Marlin Clownfish, Nemo (Marlin's son with one fin shorter than other), Dory the Regal Tang with short-term memory loss and all other characters won the heart of every animation film lover of all ages. The film presents a fully realized underwater world with bright & attractive colors and very natural dynamics - fish/tortoise movements, hydraulics, underwater illumination effects, rigid body dynamics (like the boat movement), underwater explosions, etc. The story structure is excellent with seamless connectivity among all scenes. The film is produced by Pixar Animation Studios & Walt Disney Pictures, written by Andrew Stanton, and directed by Lee Unkrich & Andrew Stanton. The film grossed about $864.62 million worldwide in 2003 which is one of the largest revenue any animation film ev er made. Pixar Animation Studios have many such successful 3D animation films at their credit. The primary process that they follow comprises of fourteen steps:- Story Idea is Pitched, Text Treatment is carried out, Storyboards are drawn (sketches), Voice recording is carried out, the virtual reels are created on the software, the artists create the look & feel, 3D Modeling is carried out, the sets are created (using computer graphics), the shots are laid, animations & behavioral aspects are added, the sets & characters are superimposed with appropriate shading, lighting of the scenes is carried out, the final computer data is rendered and finally, the finishing touches are carried out.. They were discussing their eggs when a barracuda attacked and killed Coral and ate all the eggs except one. The only egg that survived got partially damaged as a result of the attack and Marlin promises that he will never leave it - and named it as Nemo as per the wish of his wife before death. The following scene is the introduction of the film - by Pixar Animation Studios & Walt Disney Pictures.  

Healthy Lifestyles Assignment Example | Topics and Well Written Essays - 750 words

Healthy Lifestyles - Assignment Example ecifically, the detrimental consequences of these unhealthy behaviors manifest when two or more habits of the behaviors merge to produce synergistic effects. Technically, frequent health habits coalesce to form health lifestyles. Based on the interview, relevant data and information were obtained regarding particular unhealthy lifestyles. Admittedly, trends on unhealthy lifestyles suggested by the interview’s results were a bit surprising. Prior to the interview, I perceived the behaviors of tobacco use, alcohol consumption, and lack of physical exercises as experienced by members within the age bracket of 45-60 years. Surprisingly, these unhealthy behaviors are prevalent among members of the young generation; particularly those between 20-35 years. Actually, this optimally productive portion of the population seems to frequently engage in risky health habits more than those within the age bracket of 45-60 years. For example, alcohol abuse and reckless driving topped the list of most common unhealthy behaviors examined during the exercise. Observably, certain unhealthy behaviors are primarily associated with members of a particular age group. For example, teenagers and young adults between the ages of 15-24 are fond of smoking, drinking alcohol and using phones while driving compared to members of any other age group. Contrarily, adults between the ages of 55-64 years emerged as having unhealthy physical activity habits coupled with improper nutrition. With respect to teenagers and young adults, the pronounced frequency and intensity of drug abuse are relatively high compared to any other age group. In addition, reckless driving habits are more common among teenagers and young adults compared to any other age group. Reasons for this trend in young persons may include but not limited to emotional immaturity, juvenile delinquency, and social anxiety among others (Burnham, 2010). Contrarily, drinking, smoking and reckless driving habits subside towards adulthood.

Analyse using stat Assignment Example | Topics and Well Written Essays - 1500 words

Analyse using stat - Assignment Example Compare and contrast the findings from the histograms and from the tests with level and logarithmic specifications. H0: sample is not distributed normally H1: sample is distributed normally Decision: Probability value is less than 0.05significance level. Therefore we reject the null hypothesis and accept the alternative hypothesis. i.e. price variable is normally distributed Probability value is less than 0.05significance level. Therefore reject the null hypothesis and accept the alternative hypothesis. i.e. log price variable is also normally distributed Thus, the results of histogram and Kolmogorov-Smirnov test are consistent. Variable Description Combined K-S Value 1. dist Weighted distance to 5 employment centers 0.000 2. ldist Logarithm of weighted distance to 5 employment centers 0.016 H0: sample is not distributed normally H1: sample is distributed normally Decision: Probability value is less than 0.05significance level. Therefore reject the null hypothesis and accept the alte rnative hypothesis. i.e. dist variable is normally distributed Probability value is less than 0.05significance level. Therefore reject the null hypothesis and accept the alternative hypothesis. i.e. log-dist variable is also normally distributed Histogram showed the distribution of dist variable as skewed to the left. However Kolmogorov-Smirnov test yields a normal distribution. ... Mean value of rooms in the given sample is 6.28. Accordingly there are 278 houses with number of rooms bellow the average (sample A) and 228 houses with number of rooms above the average (sample B). Hence the total observations in two samples are different. Therefore we have to conduct unpaired two sample t-test. Mean Difference -8918.208 t-statistics -12.3611 P-value H0: diff=0 0.0000 H1: diff 0 1.0000 Decision: Probability value of H0 is less than 0.05 significant level. Therefore reject H0 which states there is no statistically significant difference between the mean price of houses having rooms less than average and more than average. (ii) houses below and above the average value for nox; Average nox value of the given data set is 5.549. There are 292 observations in the above average category while there are 214 observations in the below average category. Therefore unpaired, two-sample t-test with equal variances can be used. Mean Difference 6199.578 t-statistics 7.9261 P-value H0: diff=0 0.0000 H1: diff 0 0.0000 Decision: Probability value of H0 is less than 0.05 significant level. Therefore we reject the null hypothesis which states mean price of houses which are situated in lower nitrous oxide levels are not statistically different from those houses situated in higher nitrous oxide areas. (iii) Houses below and above the average value for crime. Average number of crimes committed per capita is 3.611in the given data set. Accordingly there are 128 and 378 numbers of observations in the above and below average categories respectively. Therefore, unpaired, two-sample t-test with equal variances can be used. Mean Difference 8471.173 t-statistics 9.8062 P-value H0: