Digestive System Essays (319 words) - Digestive System, Medicine

Digestive System The digestive system is a alimentary canal; it consist of the mouth, pharynx, esophagus, stomach, small intestines, large intestines and anus. The mouth is the first process of the digestive system. The mouth breaks down the food you consume. You chew the food, which is known as masticating, the food is then mixed with saliva broken down for your body to absorb and use. The food then travels to the esophagus which is connected to the stomach. contractions known as peristalsis, push the food down to the stomach. The stomach is a sac-like organ with strong muscular walls. It is located on the left side of the abdominal cavity. The food enters at the cardioesophageal sphincter. The stomach acts as a storage tank for food. It doesn't only hold the food but it also mixes and grinds the food done by chemical breakdown of proteins . The stomach is the body's major digestive organ, the nutrients are absorbed by the blood in the organ. The processed food is now chime which is then sent to the small intestines. The small intestines consist of three parts, the duodenum, jejunum, and the ileum. The duodenum is attached to the stomach, and curves around the head of the pancreas. The jejunum is attached to the duodenum and the ileum extends from the jejunum to the large intestines. The breakdown of the chyme are mixed with enzymes, and bile enters from the gall bladder. The large intestines then eliminates indigestible food from the body as feces. Though the large intestines does not participate in digestion of food, and to help pass the feces, goblet cells produce mucus to act a lubricant. The large intestines consist of the cecum, the appendix which hangs from the cecum. The large intestines leads to/consist of the colon, rectum, anus. The digestive system is used for the break down

The 8 Best Jobs for Teens and How to Find Yours

The 8 Best Jobs for Teens and How to Find Yours SAT / ACT Prep Online Guides and Tips Looking to gain a little more independence and real-world experience? Hoping to develop some useful professional skills? Getting a job as a teenager is a great way to work towards these goals. In this post, I'll talk a little bit about why you should (or shouldn't) get a job, before telling you what you should be looking for in your first part-time position. Read to the end for tips strategies on finding the best job for you! Why Should You Get a Job as a Teen? Maybe you're still deciding whether you want to jump into the world of employment; maybe you've made up your mind that getting a job is definitely right for you. No matter where you stand, I hope you read this section carefully - if you get a job for the wrong reasons, you might end up being counter-productive in the long run. Many students think it's important to get a job for the sake of their college applications. It's great to demonstrate that you can juggle different responsibilities and that you participate in activities outside of school. Keep in mind that admissions officers would rather see strong academic performances than mediocre performance + a part-time job. For this reason, I think jobs can be really great during the summer when you have a lot of free time - you can gain employment experience while also bringing in your own personal income. Working part-time while school is in session is a different story. Many students do really well in school with a part-time job, but there are more variables to consider. Namely, you have to think about prioritizing your other day-to-day responsibilities: school and homework, extracurricular activities, family, and friends. If you can balance these priorities in addition to a part-time job, go for it! If you're struggling to keep up your grades and other activities, however, you'll benefit much more in the long run by focusing on academics than you would by working part-time. It's important to find that balance between school, work, and fun. One important exception to this reasoning is if you need to get a job for financial or familial reasons. If circumstances necessitate that you work part-time while you're in school, and this negatively impacts your grades, it's important that you explain these circumstances on any college or scholarship applications. Now that we've got that out of the way - on to the fun stuff! What Should You Be Looking For in a Teen Job? Chances are you won't be making a ton of money as a teen in an entry-level position. Everyone has to start somewhere, though, and if you're thoughtful about what sort of job you want, you'll be able to get a lot out of your first employment experience. I've come up with a list of skill development areas that should generalize to many jobs (and other aspects of your life as well). You don't have to look for a job withall of the qualities I'm about to list, but here are some possible job skills benefits to consider. Think about which job qualities are most important to you: Responsibility Will people be depending on you? Is it important that you adhere to a regimented schedule, or that you complete a series of tasks or projects in order for things to function smoothly? If yes, you can expect that a particular job will help you develop a strong sense of responsibility, which is a great quality to develop as a teen. Now, most jobs will require you to demonstrate some degree of responsibility. What will ultimately vary among jobs is what you're supposed to be responsiblefor.Here's a general guide to the responsibility hierarchy that you'll find applies to most jobs: Responsible for listening to directions completing tasks Responsible for yourself - that is, doing your job without being told Responsible for taking initiative - anticipate what needs to be done, complete tasks effectively without explicit direction Responsible for other people - direct, teach, and train others. In most entry level positions, you'll start off at the lower end of this hierarchy. Some people prioritize opportunities to gain more responsibility and autonomy - do you? People Skills People skills development is a great advantage of customer service jobs (think working in a restaurant or retail store).Anyone who’s worked in customer service can tell you that you’ll encounter less-than-pleasant people, but you'll also have awesome one-on-one experiences with great customers. Any future jobs or career options will appreciate that you can work well with people, even (especially) difficult individuals. Teamwork Collaboration Some jobs are pretty solitary - you might be interacting with others in a service capacity (for example, as a checkout clerk), but you won't be collaborating with your coworkers much. Jobs where it is important to develop a sense of community, or where you work with coworkers on projects, help build your teamwork skills. Jobs that emphasize collaboration and community can be especially valuable for your resume and college applications. You'll be hard pressed to find a future employer who doesn't highly value a team player. Leadership Growth Potential You’ll find leadership growth potential in positions where you get more responsibility and/or autonomy once you’ve proven yourself. For example, in a restaurant, there might be opportunities to work your way up from busboy to waiter. In a retail environment, you could move from sales associate to shift manager. This upward trajectory comes from hard work and a demonstration of initiative, and usually accompanies pay raises and a fancier job title. This will help you both in future job searches and in your college applications. Additionally, you'll sure get a healthy dose of self-satisfaction that comes with being rewarded for your hard work. Professional Development This one might be a bit more difficult to find in an entry-level job for teens, but it’s definitely possible. By professional development, I mean developing interests, passions, and network connections based on your future career interests in a particular field. For example, if you’re interested in going to veterinary school, you might seek out work or volunteer positions at local animal shelters. If you’re interested in the hospitality industry, you could look for hostess jobs at local restaurants or front desk positions at hotels or inns. If you’re interested in research, check out labs at local universities for part-time research assistant positions. The possibilities are endless and really depend on your own unique interests and skills. 8 Great Jobs for Teens and Where to Find Them Now that we've gone through important job characteristics, I'll list some ideas for the best jobs for teens, along with corresponding job skills strategies. This list is by no means exhaustive, however, so if you have your own ideas about part-time jobs you'd like, I encourage you to do your own research and check them out! Don't worry if you have to send out many applications before getting a call for an interview; that's pretty typical for a job search. Babysitting Job skills: Responsibility, people skills How to find positions:It's best to start with families you already know. Your business will grow through word of mouth if people are happy with your services. You can also post fliers at schools, churches, and community centers. Animal Shelter Worker Job skills:Responsibility, teamwork collaboration, possible leadership growth potential, possible professional development How to find positions: Visit your local animal shelters or rescues and ask for part-time employment or volunteer applications. Some shelters may require you to be 18 or older, so you may have to ask for parental permission if you're underage. Landscaper Great summer job option, and it tends to pay pretty well (although it's very hard physical work). Job skills: Responsibility, teamwork collaboration, possible leadership growth potential How to find positions: Visit local landscape companies ahead of the busy season (spring/summer) to ask if they're looking for seasonal help. Grocery Store Clerk The great thing about clerk positions is that you can find them almost anywhere. Some larger companies even offer college scholarships (Wegmans, for example). Job skills: Responsibility, people skills, leadership growth potential How to find positions: Check online on stores' websites, or visit their customer service desks to ask for applications. Restaurant Staff Depending on your past restaurant industry experience, you might start off as a busser, waiter, or host. Bussers, in particular, don't have to be experienced, but once you're in you can work your way up the ladder. Hourly wages are really low (below minimum wage), but you can make quite a bit of money in tips depending on where you work. Job skills:Responsibility, people skills, teamwork collaboration, possible professional development How to find positions: Check online for local job postings, or ask for applications directly from restaurants. Sales Associate There are so many different types of sales associate positions; if you're in the right geographical area, it's possible to find something tailored to your interests. Are you passionate about fashion? Check out department stores or local boutiques. Are you active or outdoorsy? Look into sporting goods stores. More interested in tech? Check out computer or cell phone sales. Job skills: Responsibility, people skills, leadership growth development, possible professional development How to find positions: Check online for local job postings, or ask for applications directly from stores. Tutoring This is a great way to hone your own skills while helping someone else with your talents. Job skills: Responsibility, people skills, leadership growth potential How to find positions: Check if there are any tutoring programs offered through your school. Look into local tutoring agencies or volunteer groups. Ask your parents if they have friends with younger kids who could use tutoring help. List yourself on online tutoring platforms like Wyzant. Go Independent With newer online platforms, it's easy for budding entrepreneurs to advertise their wares and services both locally and globally. If you have a particular skill that you're passionate about, you don't have to abandon it to pursue a more typical part-time job. If you can provide a service (for example, maybe you're a graphic designer or programmer), you could likely find independent part-time work. If you're an artist or craftsman, you might be surprised to find that there's high demand for high-quality, customized goods. Job skills: Varies How to find work: Offer services through word of mouth to local businesses, friends, and family. You can also check out online platforms like Elance. If you produce goods, consider attending crafts or art fairs, or making an Etsy or Ebay account. What's Next? Looking to save your hard-earned earnings from your part-time job? Check out our guides to saving money on the SAT and ACT. If you hope to get a job so that you can save up for college, you might want to look into some of our guides to financial aid, including information on the Pell Grant, Direct Subsidized, and Direct Unsubsidized loans. Want to improve your SAT score by 160 points or your ACT score by 4 points?We've written a guide for each test about the top 5 strategies you must be using to have a shot at improving your score. Download it for free now:

Criminal Law and Historical Focus Essay Example | Topics and Well Written Essays - 2500 words

Criminal Law and Historical Focus - Essay Example (Landau, 2002) In presenting England's criminal law, Blackstone a common lawyer of earlier generation, chose not to highlight settled procedural distinctions and arcane terms, but instead he invoked a distinctive kind of legal wrong he identified as "public" in nature. The terms crime and criminal law, while enjoying wide linguistic currency, were not always the part of technical vocabulary of the law. In the medieval period, English law was explicitly differentiating between civil and criminal materials. And as is immediately disclosed by the titles of such works as Lord Kames "History of the criminal law" (1758) and William Eden's "Principles of penal law" (1771), eighteenth-century jurists certainly supposed there existed a general category of law that might serve as the object of their scholarly attention. English law, according to a familiar complaint, simply lacked much by way of system or coherent organization, particularly as compared with Roman law, which hitherto dominated university law studies and which set the relevant standard for juristic elegance. "It was thought impracticable to bring the Laws of England into a Method", explained by one of Blackstone's eighteenth-century precursors, "and therefore a Prejudice was taken up against the study of our Laws, even by Men of Parts and Learning". On this basis, English law along with the Canadian law was unable to become an object of rational learning, and instead had to be mastered through the practical, craft-like techniques of legal apprenticeship. Blackstone, as a lawyer was confident of the English law's credentials as a rational system, but the efforts of other lawyers were not considered "failure" to bring changes in the system after negotiations. Law Reform Law reform was supposed to be the one and only asset, exclusively the preserve of lawyers, and Bentham's combination of the wisdom of the reformer and the craft of the lawyer recognized its potential to divide the legal profession. The common lawyer's innate respect for existing institutions and his appreciation of the subtleties of the status quo not only made novelty in the early years of entering into the criminology, but also it particularize the concept of jurisdiction in the early years of reform, as an object of suspicion. The means and scope of amendment were a further cause for division, even after its need was established. There was a disturbing element of self-examination in law reform to which some lawyers found it difficult to respond with enthusiasm. The status quo would always find friends in influential positions, particularly in respect of criminal law reform amongst the judiciary. Law - A Profession towards Criminal Trial Process As the pace and scale of law reform increased it became an intense preoccupation of the legal profession and for the eighteenth century lawyers, acted as a charm to enter into such a jurisdiciary profession. It was the dominant theme of professional literature of that period that lasted the debate over the role of counsel and advocacy in the criminal trial, for a longer period of time and consideration in this work, could be traced in parliamentary debates, select committee minutes, the evidence and reports of the Criminal Law Commissioners, periodical and professional journals, biographies and other professional lite

Accounting Course - One.Tel Telecommunication Company Essay

Accounting Course - One.Tel Telecommunication Company - Essay Example Finally, by March 2001, One Tel was declared insolvent, after continuous efforts by News Limited and PBL to inject additional cash turned out to be insufficient. In this report, the reasons for the collapse are analyzed, how it could have been prevented and its effect. This paper analyses how the company was mismanaged, its internal control weaknesses, and financial reporting issues. Jodee Rich and Brad Keeling established One Tel. Company (telco), on the 1st of May 1995. James Packer and Lachlan Murdoch were the initial backers, providing an investment worth $995 Million (Cook, 2001). The company’s business was to provide communication services and networks in Australia and major countries of Asia and Europe. The company launched its GSM services network in 1995 and within few years it became the fourth largest telecommunications company of Australia. Its success was mainly fuelled by its marketing and ability to attract young mobile users. It achieved an early success in Australia and very soon its operations went across international boundaries. The marketing strategy was to focus on the younger generation, using the slogan â€Å"You’ll tell your friend about One Tel†. In its early years, One Tel enjoyed a lot of success due to its growing customer base and its profits grew from $3.7 million in 1997 to $6.9 million in 1999. In November 19 99, One Tel. had a market capitalisation of $3.8 billion that made it rank as the 30th largest Australian company (Barry, 2002). Unfortunately, due to gross mismanagement the company’s collapse was inevitable. In 2000, the company’s financial position reported a loss $291.1 million. The shareholders tried to keep the company afloat, but it was not to be and by 28th May 2001 the company was de-listed. The downfall of One Tel can be largely attributed to its shareholders and

FAA inspections on Boeing 737's and how it relates to Operations Research Paper

FAA inspections on Boeing 737's and how it relates to Operations Management - Research Paper Example It provides employment opportunities to over 170,000 individuals throughout 70 nations. It is also regarded as one of the oldest manufacturers of commercial airlines having its root from the last 40 years. It also manufacturers missiles, defense systems and satellites and is a major service provider to NASA (Boeing, 2013). The main aim of this research paper is to provide a broad description on the main reasons behind the Federal Aviation Administration (FAA) to order inspections of more than 1,000 Boeing 737 jets and to show appropriate reasons that the inspection of the airline can help the company in ensuring proper management of operations. Operation Management Operations management is considered as one of the most vital parts of an organizational life cycle irrespective of the fact that it is involved in producing goods or services or is functioning in private or public sectors. The main purpose or aim of operations management is to implement steps that may help the organization to operate efficiently. With reference to the article, â€Å"FAA Orders Inspections on Boeing's 737 Aircraft† by Andy Pasztor, it can be apparently viewed that the potential effect of factory defects in the manufacturing process of Boeing 737 has led the company to inspect more than 1000 Boeing 737s, causing an incurrence of total compliance costs of nearly US$10 million (Pasztor, 2013). The main steps which are usually followed in the process of operations management are discussed below: Contact customer: The first step in operations management is to obtain information on the demand, intension and the core customers/potential customers to buy the products’ produced. Moreover, conserving the contact information is also done to measure the market action (Mahadevan, 2009). Contribute a demand: The next step in the process is to propose a demand of service or product to the qualified customers. This can be done by offering a written proposal to potential customers. It is considered that business proposal is often determined to be a step in settling complex business processes. For instance, companies like Boeing usually adopt the policy of providing business proposals to potential airlines in order to offer them a broad description of the services they render and the way in which they differ from their competitors (Mahadevan, 2009; Neely, 1993). Sale: In this step, the contract or sale of the product is done to their potential customers. Establish a written procedure: In this vital step, both the parties agree upon a mutual agreement in keeping with the different aspects such as characteristics and date of supply of the final products among others. The step is recognized to be a vital part as both the parties are benefited from it (Mahadevan, 2009). Required resources: In this step, all the necessary equipments, products and additional resources needed to complete the project are verified. The costs of the entire requirements are measured to impleme nt it in inventory management (Mahadevan, 2009). Execution: In this phase, the written process is applied in the practical form, resulting in the formation of the defined product (Mahadevan, 2009). Effectiveness: The final expectation according to the contract or customer is measured. Delivery: This step involves the process of delivering the final product to the end customer. This step needs

Hydrolysis of Phosphomonoesters Study

Hydrolysis of Phosphomonoesters Study Introduction Site-directed mutagenesis was the primary basis of this portion of the laboratory directed toward determining the active site of the enzyme catalysis for hydrolysis of phosphomonoesters. The criteria was to turn the original sequence with Arginine 166 into Glutamine 166 through a point mutation. The primer location is located in the 401511 sequence of the phoA gene of alkaline phosphatase in E. Coli. The primer sequence is: 5- G CTG GTG GCA CAT GTG ACC TCG CGC AAA TGC TAC GGT CCG AGC -3’. The mutated primer sequence is: R166Q1 5’-G CTG GTG GCA CAT GTG ACC TCG CAG AAA TGC TAC GGT CCG AGC-3’. The reverse complementary mutated primer sequence is: R166Q2 5’-GCT CGG ACT CGG ACT GTC GCA TTT CTG CGA GGT CAC ATG TGC CAC CAG C-3’. A calculation for the melting temperature can be determined by the following equation: TM= 81.5 + 0.41 (%GC) – 675/N- % mismatch The TM value for Gln-166 was determined to be 85.95ËÅ ¡C. It is necessary for the TM value to be above 75ËÅ ¡C for successful mutation. Running on the null hypothesis, it would be clear that nothing would change for the kinetic parameters. If something were to change, the kinetic parameters for the Michaelis constant (Km), which determine the velocity of an enzyme as well as calculated to be  ½ the Vmax, would be hypothesized much higher after the mutation to glutamine since the mutation allows for 61% more accuracy from arginine’s CGG at 25% to glutamine’s CAG at 86%. The increase in the binding affinity creates the hypothesis that since it is bound closer together creating a stronger binding affinity, the maximal speed (Vmax) would be reduced since it does not require as much conversion between substrate and product. Mechanism of Alkaline Phosphatase Alkaline phosphatase (APase) is an enzyme located in the periplasmic space of E. Coli. The structure of APase, usually in the form of a dimer, shows two zinc metallonzymes and a magnesium ion in the active center. There is a bridging ligand with the protein Asp51 in the active center for the two zinc ions and the one magnesium ion. There are four phosphate oxygens; two phosphate oxygens form a phosphate bridge between the two zinc ions and the other two form hydrogen bonds with the guanidinium group of arg-166 (Coleman, 1992). The dimer is two identical subunits that each contain 429 amino acids. (Coleman, 1992). The most important amino acids located at the active site of APase are the four Cysteine residues represented as combined intrachain disulfides. The phosphorylated residue was Ser 102, which is phosphorylated during phosphate hydrolysis, to begin the nucleophilic attack on phosphorus. Each monomer contains a leucine rich environment of 22 residues. (Coleman, 1992). The first zinc ion is necessary in activating the leaving group of the ester oxygen in order for Ser 102 to phosphorylate (Coleman, 1992). The second zinc ion is necessary for the ester oxygen of the serylphosphate to begin activation of the phosphoseryl intermediate during hydrolysis of Ser 102. When the phosphoseryl intermediate is formed during the first zinc formation, a hydroxide is formed. (Coleman, 1992). Figure 1: this figure represents the monomer of alkaline phosphatase showing the zinc triad with magnesium in the active center. (Coleman, 1992). The proposed mutation as discussed, is mutating the arg-166 to glu-166. It is speculated that there will be no effect on the enzyme during the mutation. Arg-166 is located in the guanidinium group and is an electrophilic species while APase is being phosphorylated. Arg-166 is speculated to play a role in stabilizing the developing negative charge on the oxygen of the leaving group, pentacoordinate transition state, or help bind the phosphate group. As discovered byButler-Ransohoff et al, it is discovered that Arg-166 has no effect on the hydrolysis of phosphate monoesters and therefore will not be effected after mutation. Competent Cells Competent cells are used to help the DNA get into the cell. DNA is negatively charged and the calcium ions from the competent cells of DH5-ÃŽ ± cells are positively charged. The purpose of calcium chloride’s positive ions is to create an equilibrium between negative charge on the cell membranes phospholipid heads and the phosphate group on the plasmid DNA. This is where the genetic modification occurs. Therefore, they help by binding to the DNA and move it into the cell after the cell goes through a heat shock cycle. Competent cells have a very high rate for transformation efficiency. The proposed understanding of how this works, is that the calcium ions are positive and therefore weaken the electrostatic repulsion, which in turn weakens the cell walls. When the cell goes through heat shock, it causes the pressure to increase opening the cell and allowing the competent cells to bring the DNA into the cell by allowing pores to be created with the weakened cell wall. This forces the plasmid to become supercoiled so that it can pass through the pores created by the charge difference and heat shock. The competence comes from the cold bath of DH5-ÃŽ ± cells in calcium chloride which shocks the cells causing pores and then heat the cell to 42ËÅ ¡C for roughly 2 minutes, more than this could cause the cell membrane to denature and the pEK-154 mutated plasmid would become denatured. DH5-ÃŽ ± is a strain from E. Coli commonly used in laboratory practices due to it having the phoA- gene deficiency. This is necessary and desirable because it is easily transformed for creating competent cells. DH5-ÃŽ ± cells are used because they are a common strain in E. Coli and are commonly used for cloning. In addition, they promote stability and help improve the quality of the plasmid when using the Miniprep kit. (Dagert, 1979). A growth curve was created after incubation of the cells for competent cells. This was done to show the growth period during the lag phase is depicted by the highest point on the growth curve. This is taken to determine how much growth you have and is determined through a logarithm vs. time on a graph. The method used is optical density (OD) at 600nm to measure the transmittance on a spectrophotometer. If the value for the OD600 is high, the protection factor by a filter is lower and vise versa. This method is used to determine how much light is absorbed through the bacterial cells. Site-Directed Mutagenesis The sequence chosen for the point mutation from Arginine 166 to Glutamine 166 was sequenced in Ann Arbor. Site-Directed Mutagenesis synthesizes two complimentary oligonucleotides double stranded DNA template with glutamine 166 mutation, which is tagged with unmodified nucleotide sequences to a single stranded DNA template to allow mutation to occur and then reforms the new double strand DNA template during thermal cycling. When this occurs, the newly single stranded DNA template forms complementary strands with the enzymes and nucleotides to produce a higher result of strands. The purpose of site-directed mutagenesis is to make specific changes to the DNA sequence of a gene. Quik Change Lightening Mutagenesis kit provides a faster and more reliable insertion of the mutation with a simple three step method. The multiple cycles are subjected to a polymerase chain reaction (PCR) method. This allows multiple sets of mutated DNA to be produced within a short period of time. PCR works by heating and cooling the samples, since the double DNA is subjected to separation when heated it causes the srtands to separate allowing nicks in the template to occur. pUC-18 is a control plasmid used for determining if a mutation was successful and efficient. Figure 2: pUC-18 plasmid DNA genetically mutated to contain the LacZ gene as well as the ampicillin resistance gene. The polylinker cuts the section of the DNA by the Dpn1 restriction endonuclease creating a linear DNA strand allowing for the binding of the mutation of Arg-166 to Glu-166 to occur. pUC-18 is a circular double stranded DNA molecule. The reason pUC-18 was chosen as the control positive plasmid is because it was genetically manipulated to contain an ampicillin resistance gene as well as a ÃŽ ²-galactosidase enzyme known as LacZ. The lacZ gene is essential for DNA mutation because it contains a region to insert a polylinker, which recognizes the Dpn I restriction endonucleases during digestion causing the plasmid to become linear and bind to the mutated plasmid DNA that has also been cut with the Dpn I restriction endonuclease. A pWhitescript is used as the positive control plasmid used for mutagenesis. The control plasmid is combined with pEK154 plasmid containing the phA gene of APase and the vector double stranded DNA. The two oligonucleotide primers are the opposite ends of the vector. pWhitescript has the stop codon TAA inserted that stops the ÃŽ ²-galactosidase enzyme from producing. The two oligonucleotide control primers create a point mutation that turns the T residue of the stop codon to the C residue of the glutamine codon (Gln, CAG). That allows enzyme ÃŽ ²-galactosidase to be produced after pWhitescript is subjected to mutagenesis. There are two oligonucleotide with the mutation primer as discussed earlier. New DNA polymerase called Q1 enzyme was used for double stranded DNA template when combined with dNTP mix it extended the two-oligonucleotide primers as well as allowing the new plasmid DNA to construct. PCR Cycling parameters were used during Quik Change Site-Directed Mutagenesis for the transformation of arginine to glutamine by the use of the Pfu Enzyme. As visible from table 1, the cycling parameters were set up for this specific reaction for a point mutation. After cycling, the non-mutated parental supercoiled double stranded (dsDNA) is digested. Table 1: Quik Change Site-Directed Mutagenesis Parameters Segment Cycles Temperature Time 1 1 95C 2min 2 18 60C 60C 68C 20 sec 10 sec 30 sec/kb of plasmid length (3min) 3 1 68C 5 min The first step of the three step simple method for Quik Change Lightening Mutagenesis kit is Thermal cycling at 95ËÅ ¡C for two minutes. During this time, the DNA template becomes denatured allowing the primers to be subjected to heat treatments, which denature the pEK-154 template DNA strand and synthesizes the primers to extend linearly and cause nicks with Pfu Fusion-based DNA polymerase, which are then sealed by components within the Pfu enzyme blend. This polymerase allows for exact replication of original template and does not disrupt the orientation. The original mutated pEK-154 was used as the supercoiled double stranded DNA template Figure 1: during thermal cycling, the double-stranded DNA Template is nicked and subjected to point mutation for arginine 166 to glutamine 166 and then sealed with components in the Pfu enzyme blend. In the second step of the kit the template digests the parental DNA strand with methylated and hemimethylated DNA using the enzyme Dpn I endonuclease and subjected to thermal cycling for the second segment. The un-mutated pEK-154 plasmid DNA is the parental DNA while the mutated pEK-154 is the plasmid DNA with the desired mutation. With the nicks, the mutated plasmid with the two oligonucleotide primers is mixed with Dpn I endonuclease for methylated DNA that targets the sequence 5’-Gm6ATC-3’. This sequence helps digest the pEK-154 parental DNA template and has not been introduced into the methylase enyme allowing methylation to not effect the DNA and therefore will not be digested with the parental DNA strand. Similarly, the un-mutated is also digested in Dpn I restriction endonuclease to be methylated. Figure 2: Dpn 1 enzyme endonuclease digests the parental DNA template with methylated and hemimethylated allowing for single stranded DNA for transformation. Step 3 is the last cycling step where transformation of the newly mutated glutamine 166 single-stranded DNA. The DNA is added to Dpn I restriction endonuclease, which forms it into linear DNA stranded DNA duplex forming a double stranded DNA for the newly synthesized mutated DNA. After the mutated vector DNA with the primers has been nicked it is transformed into CaCl2 competent DH5-ÃŽ ± cells. This transformation must occur to repair the nicks caused during cycles to separate the DNA strands. The pWs, which is used as the control DNA is also transformed with competent DH5-ÃŽ ± cells to help repair the nicks. Transformation Control screening used X-gal and IPTG to determine if ÃŽ ²-galactosidase activity. The newly mutated pEK-154 cells are screened for alkaline phosphatase activity. This helps screen for ÃŽ ²-galatosidase enzyme in E. Coli is a Lac Z gene that codes for this enzyme. This enzyme is useful because it breaks the lactose into galactose and glucose. A Lac operon contains an operator and a promoter, which binds RNA-polymerase that starts transcription for the Lac I gene for I protein which can only bind to operator or lactose, but not both. When I protein is bound to lactose, in this case Isopropyl-1-ÃŽ ²-D-galactopyranoside (IPTG) which mimics the structure of lactoase allowing RNA-polymerase to bind to the promoter to produce ÃŽ ²-galactosidase. 5-bromo-4-chloro-3-indolyl-ÃŽ ²-D-galactopyranoside (X-gal), which is a substrate that cleaves galactose in ÃŽ ²-galactosidase to produce a blue color. If no ÃŽ ²-galactosidase is present they turn white on the agar plates. This determines if the cells contain the mutated CAA codon for glutamine instead of the stop codon TAA of the unmutated protein. 5-bromo-4-chloro-3-indolylphosphate (BCIP) is used to determine if alkaline phosphatase is present. BCIP like X-gal is a substrate that cleaves the phosphate group off of alkaline phosphatase producing a blue color.

indias safest cars :: essays research papers

How often do you wake up to the news of a road accident that killed a few people in some part of the country? Almost everyday, right? Then you quickly browse through the other news and life goes on. Every year, over 80,000 people die on Indian roads; every five road accidents leave one dead. Yet, it's just a statistic, which hardly changes our apathy towards road safety. Yes, road safety is an unpleasant, boring subject, but remember, it affects us all. How safe is the car you are driving or about to buy? How does one rate safety and who does it? We have compiled some information on a handful of cars that are available across the world and are also on sale in India, and their safety features and test results. We think you have the right to know. First, a little backgrounder. By law, all cars have to be tested and have to meet certain standards to be street legal. New Car Assessment Programmes (NCAPs) are carried out by various testing agencies in the United States, Europe, Japan and Australia amongst other countries. The European NCAP carries out the most stringent crash tests, as well as pedestrian and child safety tests vis-Ã  -vis other assessment programmes. Euro NCAP is a collaboration between governmental and automotive associations and is overseen by the FIA (Federation Internationale de l'Automobile). The money for these expensive tests comes from the EU, by which the widest possible range of models can be assessed. Euro NCAP carries out frontal offset, side impact and pole test and has a simple rating system that awards stars out of five. This rating system makes it easy to interpret safety standards in cars before people go out to buy them. Over and above this, manufacturers can go ahead and carry out further tests that are not required by legislation and choose to incorporate these additional safety features. Let's then see how the cars have fared now. The Hyundai Accent GLS got an adult occupant rating of 2.5 stars and a pedestrian test rating of 2 stars. The levels of chest injury in adult occupants was exceptionally high, and hence the Accent missed out a full third star. Moreover, this car was fitted with a driver side airbag. For cars sold in Europe, side airbags come as an option. Accents are also equipped with Isofix anchorages and are electronically assisted by ABS and EBD.