Free Essays on Making A Moral Decision

Settling on a Moral Decision In Joseph Conrad’s short story â€Å"The Tale† and James Joyce’s short story â€Å"A Painful Case† there are a few likenesses between the characters of the tales. Every story is made out of equal good exercises, which turns into the topic of the accounts. The principle characters glance back at a point in their lives and are uncertain on the off chance that they settled on the savvies decision in their individual circumstance. The lesson of every story is a moral choice may never have a correct answer. Conrad and Joyce depict an ethical exercise in a fundamentally the same as way. In every story the primary characters, the boss and James Duffy, think back on minutes throughout their life and lament the choices they have made. The leader in â€Å"The Tale† chooses a whole ships destiny by guiding them off precipices, thus the skipper of the boat and his group pass on. The leader states, â€Å"That course would lead the Northman straight on a fatal edge of rock. Furthermore, the leader offered it to him† (Conrad 17). The fundamental character of â€Å"The Tale† knows about the significance of his choice and questions whether it was the most shrewd decision. He examines with this and it impacts him on a passionate level. Toward the start of â€Å"The Tale† the ladies (who is never given a name) demands he advise a story like he used to before the war. She states, â€Å"You used to tell-your-your straightforward and expert stories very well at one time†¦you had kind of craftsmanship in the days-the days prior to the war† (Conrad 2). This announcement supplies the peruser with proof that the boss is a changed man since the war and hints that the story won't resemble his old â€Å"simple† stories. All through the story the leader fights with his impulse to choose whether or not to accept the Northman. He consistently wonders why the Northman didn't sound his horn when they entered the inlet, and why the Northman’s vo... Free Essays on Making A Moral Decision Free Essays on Making A Moral Decision Settling on a Moral Decision In Joseph Conrad’s short story â€Å"The Tale† and James Joyce’s short story â€Å"A Painful Case† there are a few likenesses between the characters of the narratives. Every story is made out of equal good exercises, which turns into the topic of the tales. The fundamental characters glance back at a point in their lives and are uncertain in the event that they settled on the most astute decision in their individual circumstance. The lesson of every story is a moral choice may never have a correct answer. Conrad and Joyce depict an ethical exercise in a fundamentally the same as way. In every story the primary characters, the leader and James Duffy, think back on minutes throughout their life and lament the choices they have made. The boss in â€Å"The Tale† chooses a whole ships destiny by guiding them off precipices, subsequently the commander of the boat and his team bite the dust. The leader states, â€Å"That course would lead the Northman straight on a dangerous edge of rock. Also, the boss offered it to him† (Conrad 17). The fundamental character of â€Å"The Tale† knows about the significance of his choice and questions whether it was the smartest decision. He mulls over with this and it impacts him on an enthusiastic level. Toward the start of â€Å"The Tale† the ladies (who is never given a name) demands he advise a story like he used to before the war. She states,  "You used to tell-your-your straightforward and expert stories very well at one time†¦you had kind of craftsmanship in the days-the days prior to the war† (Conrad 2). This announcement supplies the peruser with proof that the leader is a changed man since the war and hints that the story won't resemble his old â€Å"simple† stories. All through the story the boss fights with his nature to choose whether or not to accept the Northman. He persistently wonders why the Northman didn't sound his horn when they entered the narrows, and why the Northman’s vo...

Myself Included

Myself Included Myself Included Myself Included By Maeve Maddox A peruser questions my utilization of the expression â€Å"myself included† in the accompanying concentrate from a post on who versus that: Numerous speakers, myself notwithstanding, feel that who is generally the main decision when the predecessor is human, yet perceive that its utilization is an elaborate decision and not a matter of rule. Now and again that might be the better decision. Says the peruser: I’m pondering about your utilization of ‘myself’. I would have expected it ought to be ‘me included’. Or on the other hand have I misconstrued the reflexive pronouns? I’m inquisitive in light of the fact that it’s one of my annoyances when somebody closes an email with ‘if you have any inquiries, you can allude to myself.’ The peruser has not misconstrued the general standards for the reflexive pronouns. 1. A reflexive pronoun is utilized as an immediate article when the item is equivalent to the subject of the action word: â€Å"I cut myself shaving again.† 2. The reflexive pronoun is utilized as a roundabout article when the aberrant item is equivalent to the subject of the action word: â€Å"She got herself another car.† 3. The reflexive pronoun is utilized as the object of a relational word when the article alludes to the subject of the proviso: â€Å"My child manufactured our deck by himself.† Note: The expression â€Å"by + reflexive pronoun† shows that somebody accomplished something alone and additionally with no assistance. A similar significance is passed on when the reflexive pronoun alone is set toward the finish of a sentence: â€Å"I heated all the treats myself.† 4. The reflexive pronoun is utilized to underscore the individual or thing alluded to: â€Å"The restricting itself is worth  £50.† Note: This utilization of the reflexive pronoun is particularly basic when the individual alluded to is popular or amazing: â€Å"The Queen herself composed a note of sympathy to her butler.† The most widely recognized mistakes made with reflexive pronouns are the sort the peruser alludes to, the utilization of a reflexive pronoun when the setting requires a plain close to home pronoun: Wrong: If you have any inquiries, you can allude to myself. Right: If you have any inquiries, you can allude to me. The blunder here is utilizing a reflexive pronoun as the object of a relational word that doesn't allude to the subject of the proviso (you). Other regular mistakes incorporate the accompanying: Inaccurate: Jack and myself made a trip to Greece this late spring. Right: Jack and I headed out to Greece this mid year. The blunder is in utilizing the reflexive pronoun as the subject of an action word. Inaccurate: When you give out the presents, don’t overlook Margie and myself. Right: When you give out the presents, don’t overlook Margie and me. The blunder here is utilizing reflexive myself as the object of the action word overlook. In spite of the fact that the expressions â€Å"myself included† or â€Å"including myself†seem to challenge the standards they have delighted in a long history of utilization by legitimate authors. The Ngram Viewer demonstrates that â€Å"including myself† is unmistakably more typical than â€Å"including me† in printed books. A Web look for â€Å"including me† brings 617,000 outcomes; â€Å"including myself† brings 3,890,000 outcomes. An article by language specialist James Harbeck records fourteen instances of exemptions to the standards. Here are three of them: You appear to be a superior rendition of myself. (object of relational word) There are two others here other than myself. (object of relational word) Myself, as executive here, will cut the strip. (subject of sentence) Now and then â€Å"including me† is the undeniable decision, however in different settings, an essayist may favor â€Å"including myself.† Compare: Everybody got a sumptuous blessing, including me. Numerous researchers, including myself, found the film crazy in its mistakes. The best guidance about the utilization of reflexive pronouns is to ace the standards, yet to stay mindful that occasionally â€Å"nonstandard† myself might be more colloquial than me. â€Å"If you have any inquiries, you can allude to myself† is undeniably nonstandard, yet in settings in which a speaker or author is embracing a sentiment shared by others, â€Å"myself included† and â€Å"including myself† are built up expressions. Need to improve your English in a short time a day? Get a membership and begin accepting our composing tips and activities day by day! Continue learning! Peruse the Grammar classification, check our well known posts, or pick a related post below:Spelling Test 18 Types of Parenthetical PhrasesCaptain versus Ace

Bullying at School Essay Example | Topics and Well Written Essays - 1250 words

Harassing at School - Essay Example At the point when an understudy humiliates, derides or despises another understudy it is provocation, harassing or prodding. (Harber, 2004) Harassing can take numerous structures physical brutality, dangers, verbally abusing, mockery, spreading gossipy tidbits, determined prodding, and prohibition from a gathering, tormenting, criticism, embarrassment and injurious remarks. All are a type of viciousness. Instructor understudy tormenting additionally exists. Student understudy tormenting is a typical issue in schools universally. In spite of the fact that there are varieties in the kinds of harassing executed, tormenting is done by the two guys and females and the two guys and females are the people in question. Be that as it may, clearly schools have no impact in making it and put forth a valiant effort to stop it. Schools can have an enormous impact in really making the issue of harassing. For instance, lower accomplishing students, deprecated in the serious environment, may endeavor to recover some pride through harassing. It is additionally brought up that while school sport has the potential for learning camaraderie, putting forth a strong effort and shared undertaking, it can likewise reject the individuals who don't exceed expectations and it can get brutish with over-intensity, over-focusing on the body and the advancement of animosity and viciousness. While all kids who are dealt with fiercely don't proceed to become menace in light of the fact that there are regularly countervailing and adjusting socialization impacts, and on the grounds that they don't have the chance, genuine harm should be possible to people and the outcomes can contrarily influence social orders. The second, related thought is that dictatorship and its accentuation on programmed compliance to orders is perilous as it conditions and allows people to complete vicious acts by demonstrating a defense or legitimating for them. Numerous individual demonstrations of savagery have been done and defended for the sake of the obligation to comply. Kinds of Violence Menaces ordinarily resort to different sorts of savagery they regularly subject casualties to, as physical mistreating and mental and passionate badgering. One significant part of this can incorporate different types of inappropriate behavior verbal sexual animosity, the danger of sexual maltreatment, spontaneous physical contact and upheld sexual impedance. The hidden dictator and man centric setting of direct sexual savagery in tutoring and its job in duplicating authoritative types of vicious manliness are frequently all the more harming for the harassed kids. the instructors seemed to seek after their desirous exercises both inside and outside the study hall straightforwardly; in the homeroom, young men and young ladies would whistle or murmur if an educator approached a specific young lady known to hold any importance with him to recite so anyone can hear or come before the class. Young men were noisy in their judgment of such educators, not for moral reasons but since they considered it to be uncalled for rivalry. (Harber, 2004) In addition, male educators who carry on along these lines are showing to young men that such conduct is worthy. Some key discoveries of the examination were: - Sexual maltreatment of young ladies by young ladies by male students and instructors is acknowledged alongside whipping, boisterous attack and tormenting, as an unavoidable piece of quite a bit of school life. It abuses inconsistent force connections and the

Team Ground Rules and Guidelines Free Essays

What are the general desires for all individuals from the group? Sarah Dowling-It is our objective as a group to work cooperatively to guarantee all members’ considerations, thoughts, and info are shared, examined, considered, and fused into the learning group assignments and undertakings. It is people’s obligation to be responsible for their individual segment of the undertaking and to endeavor to do their own best. As a gathering we will search out circumstances inside each other’s qualities to improve our nature of work and help those colleagues who may battle to do as such. We will compose a custom article test on Group Ground Rules and Guidelines or then again any comparable subject just for you Request Now In the occasion difference happens between associates concerning any one thought, including yet not constrained to: substance, quality, or course of events of work submitted, colleagues will cast a ballot to choose if the individual/thought will keep on working with the gathering or venture. Every part should be focused on the team’s scholastic accomplishment and achievement. Desires for Time Management and Involvement (Investment, correspondence with the group, openness, and so on.) Sarah Dowling-Our group will impart often by means of email and telephone, just as when class. Colleagues consent to go to all gatherings inâ whichever strategy conceivable. All people must finish their part of the settled upon work in an opportune way that will be dictated by the gathering all in all. Guaranteeing Fair and Even Contribution and Collaboration What technique will you use to guarantee that all colleagues are contributing and working together properly? Portray the correspondence procedure you will utilize if a colleague isn't contributing and working together effectively.How will the group oversee clashes between colleagues? Sarah Dowling-Team individuals will show up on time to gatherings and consent to impart information and progress to their individual bit of cooperation. A missing colleague acknowledges the choice of the group with respect to credit focuses recorded in the group log. It is each member’s duty to impart to the group their legit accessibility, information, and ability of finishing the gathering assignments. Techniques to determine poor work quality incorporate extra mentoring and workshops to be finished by people varying to guarantee high scholastic accomplishment as the objective of the gathering. Extraordinary Considerations What do you, as a group, concur will make this group experience not quite the same as past group encounters? Sarah Dowling-By viably conveying among one another and sharing thoughts the group will be fruitful. A guarantee to a course of events and a devotion to delivering quality work will guarantee that all group ventures are executed with a high scholarly norm. Step by step instructions to refer to Team Ground Rules and Guidelines, Papers

Oops. U.S. News Ranking Screwup

Oops. U.S. News Ranking Screwup by: John A. Byrne on September 10, 2014 | 0 Comments Comments 2,316 Views September 10, 2014Oops.U.S. News World Report, which published its new undergraduate rankings yesterday, admitted it goofed in publishing incorrect rankings in a trio of key business specialties: accounting, entrepreneurship, and finance.The errors were particularly egregious because they were so glaringly obvious. The top listed schools in entrepreneurship failed to include long-recognized Babson College as one of the leading schools for entrepreneurs. Prior to this year, Babson’s undergraduate business program has been ranked first in entrepreneurship for an unprecedented 17 consecutive times. With the new correction, the school has now made that 18 in a row.Even more embarrassing for U.S. News, perhaps, was the fact that the top of the specialty rankings for finance failed to include either the University of Pennsylvania’s Wharton School, which has for deca des been considered the best business school in that area. Nor did it  include New York University’s Stern School of Business, another prominent school known for its superb finance faculty. The University of Texas McCombs School, meantime, also long recognized for the quality of its accounting department, went completely missing from the ranking of the best schools in accounting.SPECIALTY RANKINGS ARE BASED ON A SURVEY OF DEANS AND SENIOR PROFESSORSU.S. News’ specialty rankings are based entirely on the magazine’s survey of business school deans and senior faculty, many of whom vote on the basis of a program’s reputation. U.S. News surveys two officials from each school  accredited by the Association to Advance Collegiate Schools of Business.In the spring of 2014, participants were asked for nominations of the 10 best programs in business specialty areas such as accounting, marketing and finance.  Programs  receiving the most mentions in each area are  ranked in descending order. A school or program had to  receive seven or more nominations in a specific  specialty area to be listed. This year, some  38 percent of those surveyed responded, a two percentage point improvement over last year in the response rate.But something went wrong with the way U.S. News’ calculated those peer scores. â€Å"In three specialties in the Best Undergraduate Business rankings – accounting, entrepreneurship and finance, which are based solely on peer reputation – we incorrectly loaded peer reputation scores,† said Robert Morse, chief data strategist for the rankings, in an email to the schools. â€Å"U.S. News has now loaded the correct data for these three specialties and recalculated the rankings.†UT-AUSTIN IS NOW AT THE TOP OF THE ACCOUNTING RANKINGThe top five schools in accounting are now No. 1 University of Texas at Austin, No. 2 University of Illinois at Urbana-Champaign, No. 3 Brigham Young Univ ersity, No. 4 University of Notre Dame, with Wharton and the University of Southern California tied for fifth place. The earlier released incorrect list failed to have UT-Austin in the top slot but rather Illinois in a list of 36 ranked schools.The top five schools for startup types are now No. 1 Babson College, No. 2 MIT, No. 3 University of Southern California, and two schools tied for fourth, Indiana University and Wharton. Previously, U.S. News reported that MIT had won top honors in this category among the 25 schools given a numerical rank.The top five schools in finance are now No. 1 Wharton, No. 2 NYU Stern, No. 3 University of Michigan, No. 4 MIT, and No. 5 UC-Berkeley’s Haas School of Business. Initially, U.S. News failed to include both Wharton and NYU at the top, instead listing Michigan in the lead. Wharton had been sixth on a list featuring 29 ranked schools.RANKINGS WONT GET CORRECTED IN U.S. NEWS PRINT GUIDEBOOKU.S. News said the recalculated rankings in these three areas have been updated online, but it would not be able to correct the print guidebook of the rankings because it has already been published.This is not the first time a major ranking organization has made a mistake on its list. Two years ago, Bloomberg BusinessWeek incorrectly calculated some statistics that lead to numerous changes in its MBA program rankings. In that case, the correction did not appear until nearly a year after the rankings were published. To its credit, U.S. News made the fix by 4 p.m. on the same day it issued the rankings yesterday (Sept. 9).Schools take these rankings seriously because they greatly impact reputation, application volume, alumni donations, and even the ability of an institution to attract the best faculty.  Ã¢â‚¬Å"Babsons continued recognition as the leader in entrepreneurship education and top-30 ranking in a list dominated by large, national universities reinforces the College’s leadership position in business education,† said  Scott Moore, dean of Babson’s undergraduate school, in a statement.(See the following page for U.S. News’ specialty rankings) Page 1 of 41234 »

The metabolism of methionine - Free Essay Example

Homocysteine, a non-protein amino acid, is an intermediate in the metabolism of methionine and biosynthesis of cysteine. It has gained prominence in the past half-century because its accumulation in the body has been linked to increased risk and occurrence of atherosclerosis and cardiovascular disease. Homocystinuria is due to errors in metabolism and homocysteinemia is attributed to polymorphisms in the genes involved in methionine metabolism and cysteine biosynthesis, and deficiencies in the nutrients folic acid, and vitamins B12 and B6. Proof of these can be found in the results of genetic and dietary studies. The clinical quantification of homocysteine has evolved through the years and routine tests are currently available. Baseline homocysteine concentration for normal adults has been identified to be between 12-15  µmol/L. Elevated levels of homocysteine can be reduced with folate and B vitamins supplementation, but this intervention does not work in patients who have suffered stroke or heart attacks. More research studies have supported the hypothesis that elevated homocysteine causes CVD and is not just a marker for the disease. However, despite the amount homocysteine research performed, many issues remain to be resolved, among which are the elucidation of the molecular mechanism of the direct action of homocysteine and the standardization of techniques for the quantification of homocysteine levels. Brief introduction. 200 w The role of elevated levels of homocysteine in blood plasma has been the subject of intense study and literature reviews for more than 50 years since an association between defects in homocysteine metabolism and thromboembolism was observed. This role was further verified in patients with homocystinuria and abnormalities in vitamin B12 metabolism with general vascular damage and widespread thrombosis. Subsequently, the homocysteine theory of arteriosclerosis was formulated by McCully and Wilson in 1975. Since then, the positive association between the risk of cardiovascular disease and homocysteine levels in the general population was validated in many epidemiological studies (Boushey, et al., 1995; Verhoef, et al., 1996; Eikelboom et al., 1999; Humphrey et al., 2008). Significantly, these studies found that small increases of homocysteine levels in blood increase the risk of coronary heart disease. Another important finding was the role of diet, vitamins and folic acid in lowering h omocysteine levels. Some authors have questioned the direct homocysteine-cardiovascular disease link, basing their conclusions on a review of longitudinal, prospective studies (Kaul, Zadeh and Shah, 2006). Accordingly, the evidence is not strong enough to warrant a causal effect, the mechanisms for how homocysteine causes cardiovascular disease has not been elucidated, and that there has been no proof showing that interventions of decreasing homocysteine levels have modified the risk for atherosclerosis (Kaul, Zadeh and Shah, 2006). Moreover, homocysteine has been proposed to be a marker, and not a cause of CVD (Wierzbicki, 2007). Recent studies have proposed a mechanism on increased risk of cardiovascular disease (CVD) due to elevated homocysteine. Inhibition of the growth of endothelial cells and promotion of vascular smooth muscle cell proliferation results in damage of vascular cells (Zou, 2007). The abnormality in the production of endothelial cells was caused by homocysteines inhibition of DNA methylation in the promoter region in the gene of an activator of the cell cycle (Jamaluddin, et al., 2007). Amino acid and focus on cysteine. 300 w All living cells contain the biomolecules proteins, carbohydrates, nucleic acids and lipids (Mathews and Van Holde, 1996; McKee and McKee, 2004). These are made up of the repeating sub-units of amino acids, sugars, nucleotides and fatty acids respectively. Cells also contain small organic molecules that are involved in complex biosynthetic and regulatory pathways that are tightly controlled at the molecular level. Amino acids are naturally occurring compounds containing an amino group, a carboxyl group, and a unique side chain or R group (Figure 1). The chemical and functional properties of an amino acid are largely determined by its R group (McKee and McKee, 2004; Mathews and Van Holde, 1996). Although there are hundreds of amino acids, only twenty have been identified to be building blocks of proteins. Among these is cysteine, which contains a sulfhydryl, or thiol (SH) in its R group (-CH2-SH) (Berg, Tymoczko and Stryer, 2002). The sulfhydryl group is very reactive and could react with another thiol group, forming disulfide bonds or sulfide bridges. These bridges are important in increasing the stability of some proteins. Two sulfhydryl moieties of two cysteine bonds can oxidize forming cystine. This commonly occurs in extracellular fluid like blood and urine. However, the solubility of cystine is very low, and in large amounts, cystine can cause kidney stones (McKee and McKee, 2004).The sulfhydryl moiety also forms weak bonds with nitrogen and oxygen. Another amino acid which contains a sulfhydryl group is methionine. Its side chain is -CH2CH2SCH3. The sulfur in methionine can form bonds with electrophiles (Berg, Tymoczko and Stryer, 2002). The methyl group (-CH3) can be activated and is involved in many reactions where just one carbon atom is being added to another compound. Cysteine is considered a non-essential amino acid because it can be synthesized de novo in both plants and animals (Figure 2). In animals, cysteine is formed from the cleavage of cystathionine through the action of ÃŽÂ ³-cystathionase. Cystathionine is produced when serine condenses with homocysteine that is derived from methionine (McKee and McKee, 2004). Methionine, an essential amino acid, cannot be synthesized in humans, and thus, have to be provided in the diet. Its metabolism involves homocysteine. Homocysteine structure and metabolism. 200 w Homocysteine is a non-protein amino acid and thus, it is not utilized in protein synthesis. Its structure is similar to cysteine but it has an additional carbon before the sulfhydryl moiety (R group: -CH2 -CH2 -SH). The tendency of homocysteine to form cyclic compounds may have limited its potential as a protein building block. Homocysteine is not provided in the diet, but is biosynthesized from methionine in a process involving several steps. Methionine is first adenosylated to form S-adenosyl methionine (SAM), and then the methyl group is transferred to an acceptor molecule to form S-adenosyl homocysteine (SAH) in a process called transmethylation. Adenosine is then removed thereby forming homocysteine (Figure 3) (Durand et al., 2001; Selhub J. , 1999; Marinou, et al., 2005). Transmethylation is the only pathway for producing homocysteine in the body. Homocysteine then undergoes two pathways: remethylation to methionine, and transsulfuration to produce cystathionine, cysteine, pyruvate and taurine. Remethylation requires folate and cobalamin (vitamin B12), and transsulfuration requires pyridoxine (vitamin B6). Homocystinuria results from genetic errors in the metabolic pathways (Kluijtmans, et al., 2003; Klerk, et al., 2002), but the current focus is on the more common hyperhomocysteinemia that results fr om genetic variations or nutritional inadequacy (Finkelstein and Martin, 2000). Polymorphisms in the gene methylenetetrahydrofolate reductase (MTHFR) which catalyzes the transfer of a methyl group to homocysteine to re-form methionine have been found in many studies to increase homocysteine levels (Klerk, et al., 2002). Among the proposed effects of increased homocysteine levels resulting in CVD are oxidation of low density lipoprotein, decrease in the thrombomodulin expression inhibiting the anticoagulant pathway leading to thrombosis, platelet activation and aggregation, and smooth muscle cell proliferation (Eikelboom, et al., 1999; Jamaluddin, et al., 2007). Normal Homocysteine levels. 200 w The concentration of circulating total homocysteine (tHcy) is an accurate marker of low folate and vitamin B12. In 1999, 3563 male participants and 4523 female participants were surveyed to determine the normal blood homocysteine levels (Selhub, et al., 1999; Selhub, 1999). The survey found that homocysteine increased with age and was higher in adult males than and females. Screening of babies confirmed that homocysteine levels are higher in baby boys (Refsum, et al., 2004). It was recommended that blood homocysteine levels equal to or greater than 11.4  µmol/L (males) and 10.4  µmol/L (females) are to be considered above normal and was associated with low vitamin concentrations in two-thirds of the subjects surveyed (Selhub, et al., 1999). This verified that an assessment of homocysteinemia requires the knowledge of the health status of the patient. The degree of homocysteinemia is classified based on determination fasting levels of serum homocysteine. Currently, a plasma tHcy level of 15  µmol/L is considered as normal (Refsum, et al., 2004). In healthy adults with good folate and B vitamins status, the upper reference limit is 12  µmol/L. Accordingly, homocysteinemia is classified as moderate (15-30  µmol/L), intermediate (30-100  µmol/L) and severe (100  µmol/L). This categorization is necessary to come up with a decision regarding the treatment of the condition. Homocysteine and nutrition. 300 w Studies have shown that the enzymes regulating the metabolism of homocysteine are activated by the B vitamins and folate which can be sourced from the diet. The balance of the remethylation and transmethylation pathways are coordinated by nutritional inputs, specifically methionine (Selhub, 1999). Decreasing dietary methionine increased homocysteine remethylation, while increased methionine increased the production of cystathionine through upregulation of the transsulfuration pathway. The molecular mechanisms for the effects of dietary methionine are based on the capacity of S-adenosyl methionine SAM to inhibit methylenetetrahydrofolate reductase (MTFHR) and activate cystathionine synthase (Selhub and Miller, 1992). Thus, when dietary methionine is high, there is a rapid conversion of methionine to SAM. The high levels of SAM promote the inhibition of MTHFR and subsequently, remethylation of homocysteine is also depressed. The pathway is channeled to the transsulfuration pathway, to produce cysteine. In opposite conditions of low dietary methionine, low SAM levels are not enough to inhibit MTFHR activity, resulting in the remethylation of homocysteine. The role of folate in homocysteine metabolism has been the subject of many research studies (Antoniades, et al., 2009; Boushey, et al., 1995; Lonn, et al., 2006). A diet that is poor in folate was also found to impair remethylation and synthesis of SAM (Miller, et al., 1994). Folate deficiency increased the concentration of tHcy and hepatic SAM concentration. Introducing dietary folate and methionine also decreased tHcy. Since vitamins and trace minerals are lost due to food processing, fortification is proposed in order to replenish the lost nutrients. Folic fortification of breakfast cereals when consumed daily was recommended to be the most effective means of decreasing serum folate (Riddell, et al., 2000). High-dose folic acid supplementation also decreased tHcy levels in cases where there is vitamin B12 deficiency (Min, 2009). However, a high methionine diet is not recommended since it can decrease body weights and HDL-cholesterol production in mice (Velez-Carrasco, Merkel and S mith, 2008). Aside from the observed effects of folic acid on reducing tHcy concentrations, folic acid is also thought to be involved in ameliorating endothelial dysfunction through its action of maintaining endothelial nitric oxide synthase in its coupled state which favors the formation of nitric oxide, and not oxygen radicals (Moens, et al., 2008). Role of Homocysteine and vitamins. 200 w The enzymes involved in metabolism of homocysteine require the participation of vitamin cofactors B6 and B12, and folic acid. The direct association between low folate, vitamin B6 and B12 status and hyperhomocysteinemia have been validated many times Hao, et al; 2007; (Hao, et al., 2007; Boushey, et al., 1995; Kluijtmans, et al., 2003; Koehler, et al., 2001). Investigations have geared towards the probability of using these vitamins in the prevention and treatment of homocysteinemia in the general population, and in those who already have cardiovascular disease. The use of multivitamins supplementation for several months improved the concentration of the B vitamins and folate acid in plasma, and decreased homocysteine and LDL cholesterol levels in 182 study participants (Earnest, Wood and Church, 2003). Similar homocysteine-lowering effects were also observed in patients with celiac disease, who have malabsorption problems, who take vitamin supplements (Hadithi, et al., 2009). However, the case for the vitamins is different in patients who already have CVD. Although the treatment of different combinations of vitamins B6, B12 and folic acid of patients with coronary artery disease resulted in a 30% reduction of tHcy one year after receiving vitamin B12 and folic acid, follow-up measurements of homocysteine levels showed that the vitamin supplementation did not significantly affect the total cardiovascular events (Ebbing, et al., 2008). Thus, the findings do not support the use these vitamins for secondary prevention of coronary artery disease. The Norwegian Vitamin Trial (NORVIT) also found that vitamin supplementation did not reduce the risk of cardiovascular disease recurrence after a heart attack (BÃÆ' ¸naa, et al., 2006). Moreover, preliminary results of the NORVIT suggested that the B vitamin treatment increased risk of cancer, and that such treatment should not be given. Role of Homocysteine in atherosclerosis. 500w Many studies since the 1960s have validated the relationship between hyperhomocysteinemia and risk of atherosclerosis (Eikelboom, et al., 1999). Severe hyperhomocysteinemia (homocysteine levels greater than 100  µmol/L) can be caused by several inherited genetic disorders. Foremost among the disorders are mutations in the gene encoding for the major transsulfuration enzyme cystathionine ÃŽÂ ²-synthase, remethylation enzymes methylenetetrahydrofolate reductase, methionine synthase in the methionine cycle, or defects in vitamin B12 metabolism (Finkelstein, 1998; Kraus, 1998). These genetic conditions lead to extreme elevations of plasma homocysteine and early atherothrombotic disease, where the typical pathologic features of endothelial injury, vascular smooth muscle cell proliferation, and progressive arterial stenosis are observed. While these genetic errors in metabolism are rare, they gave researchers a model for studying cardiovascular injury that was associated with hi gh homocysteine levels. The clinical and pathologic features observed under the conditions of homocystinuria resulted in the homocysteine theory of atherosclerosis that declared high plasma homocysteine concentrations to be responsible for vascular damage. Results of individual studies and meta-analysis have come up with support for the homocysteine theory of atherosclerosis. However, there are still controversy over whether homocysteinemia causes CVD or vice versa, or is it that homocysteinemia is just a marker or indicator of CVD. Mounting evidence for the causal effect of homocysteine was backed up by basic and cellular studies which utilized advanced molecular and genetic techniques. Early studies infer that homocysteine damages the endothelial cells lining the blood vessels and increase the production of vascular smooth muscle (Berg, Tymoczko and Stryer, 2002). From a pathophysiologic point of view, homocysteinemia is associated with increased oxidative stress in the cells, and development of thrombosis (Tyagi, et al., 2005), impaired endothelial function (Stuhlinger, et al., 2001) and the activation of inflammatory pathways that are sensitive to changes in cellular redox states (Weiss, et al., 2003). It was originally proposed that homocysteinemia promotes the accumulation of S-adenosyl homocysteine, a potent inhibitor of cellular DNA methylation (Zou, 2007). The hypomethylation of DNA could affect promoter activity resulting in remodelling of chromatin and changes in the transcription of certain genes. However, it was found that In the case of homocysteinemia, increased tHcy levels cause the hypomethylation of DNA promoter region of the cyclin A gene (Jamaluddin, et al., 2007; Zou, 2007). Cyclin A protein is a sub-unit of cyclin-dependent kinases (CDK) that activates CDK during the cell cycle. In the presence of high tHcy concentration, methylation was inhibited at two CpG sites in the cyclin A promoter, resulting in a 6-fold increase in promoter activity, and therefore increased proliferation of endothelial cells. Homocysteine directly inhibited the activity of DNA methyltransferase I (DNMT1) by 30% (Jamaluddin, et al., 2007). Furthermore, homocysteine reduced binding of methyl CpG binding protein 2, while it also increased the binding of cyclin A promoter to histones H3 and H4 which led to chromatin remodelling. Review methods of analysing homocysteine and differences between measurements. 2800 w Total homocysteine, tHcy, is the sum of all chemical species of homocysteine that is present in the blood (Figure 4) (Ueland, et al., 1993; Refsum, et al., 2004). As mentioned earlier, only a small portion of the tHcy comes from free homocysteine, a large portion comes from mixed disulfides and protein-bound homocysteine. All the assays used to quantify tHcy cleave and convert all these forms into a single species by adding a reducing agent, therefore what is measured is the reduced form. Blood for homocysteine determination is collected in tubes. If plasma homocysteine is to be measured, the tube contains anticoagulants like EDTA, heparin or sodium citrate. If serum homocysteine levels are to be determined, then the samples are allowed to coagulate, which takes thirty minutes and not more, to reduce the risk of increased homocysteine levels coming from the cellular fraction of blood (comprised of red blood cells and platelets). Increased release of tHcy from cells also makes it necessary to centrifuge and separate blood components within 30 minutes of collection. Centrifugation time and speed are relatively low (5 min and 300g respectively, for example) and is performed at 4 ° C. The supernatant (serum/ plasma fraction) that is collected may be analyzed immediately, or stored for 2-3 weeks in the refrigerator or frozen for 2-3 months (Homocysteine, 2008). It is possible to store the samples once processed because the homocysteine levels have been shown to be s table after separation of plasma/serum from the cellular fraction (Refsum, et al., 2004). There are reports showing that tHcy in serum can remain stable for several years if stored at -70  °C. Scheme for the steps followed in the determination of total homocysteine from blood serum or plasma. (Hcy-SR, Hcy-mixed disulfide; EC, electrochemical detection; LC, liquid chromatography; CE-LIF, capillary electrophoresis with laser-induced fluorescence detection; Ab, antibody; EIA, enzyme immunoassay; FPIA, fluorescence polarization immunoassay; MS-MS, tandem mass spectrometry). Figure from Refsum et al., 2004. The first step is to generate free homocysteine by chemical reduction of disulfide bonds using dithioethritol, sodium borohydride, n-tributyl phosphine, 2-mercaptoethanol and phosphine tris (2-carboxyethyl) phosphine, which is water-soluble (Jacobsen, 1998). After the reduction, homocysteine is separated from the cysteine, glutathione and cysteinylglycine, low molecular weight thiols, by HPLC. Homocysteine can also be derivatized for gas chromatography determination/ HPLC or subjected to immunoassay (Jacobsen, 1998; Refsum, et al., 2004). The development of the methodologies for determining total homocysteine started in the 1980s. There are many variants of the methodologies, but they can be categorized into three general methodologies: (1) chromatographic, (2) enzyme immunoassays and, (3) enzyme cycling. Chromatographic methods The earliest methods used were the chromatographic methods, of which high pressure liquid chromatography (HPLC) is more used compared to gas chromatography (GC). Variations in the methodology include the combination of liquid chromatography (LC) with mass spectrometry (MS) and GC with MS. The equipment and the skill requirements for these techniques have limited their use mainly in research laboratories. In the HPLC method, the reduced samples are deproteinated and then directly separated on an ion exchange column. The separated homocysteine molecules undergo derivatization with ninhydrin (a chromophore for colorimetric quantification), or fluorescent compounds (e.g. fluorescamine, SBD-F). Alternatively, the reduced homocysteine molecules are first derivatized before separation through reversed-phase HPLC. 6-Aminoquinolyl-n-hydroxylsuccinimidyl carbamate (AQC) is a fluorophore that gives higher sensitivity when used to derivatize cystathionine and homocysteine (Seo, 2005).The derivatized samples are then passed through an ion exchange column. The length of time that it takes for the sample to go through the column is called the retention time. The samples leaving the column pass through a detector, which could be a UV absorbance detector (spectrophotometer), a fluorescence detector or a mass spectrometer which is a powerful detector. The detectors quantify the samples, and send signa ls to a recorder which produces the chromatogram, where quantities can be seen as peaks. An internal standard of known quantity is used in the determination of the actual quantity of the sample. Normally, the internal standard should have a retention time that is the same as homocysteine. Internal standards are added to the sample tubes and derivatized together with the reduced homocysteine. To quantify homocysteine, the tHcy peak area is compared with the peak area of the standard eluted at the same retention time. Although there are many advancements in the HPLC technology, tHcy quantification using this method is relatively slow, time-consuming, requires very careful sample preparation and handling, and skilled technicians. Sample pre-treatment requires 30-60 minutes, while an HPLC run takes 10-30 minutes. Samples are run sequentially, which makes HPLC not suitable for high throughput determinations. Some laboratories use gas chromatography or gas chromatography with mass spectrometry to get more sensitive results. The principles behind GC and HPLC determination are similar, except that in GC, the samples are vaporized and passed through a gas before elution from a column. Similar to HPLC, GC is also more time-consuming compared to other methods. Homocysteine of dried blood samples have been determined using reversed phase HPLC (Bowron, et al., 2005). In this technique, blood samples were collected in tubes that contain potassium EDTA. After mixing gently, the blood was spotted on filter paper cards that are used in standard neonatal screening. The cards were air-dried and stored at room temperature until analysis. Six-mm discs of the cards were punched out, and incubated in tubes with an internal standard and a deproteinization agent. The produced thiols were then derivatized for fluorescent detection, and separated by reversed phase HPLC. The resulting measurements showed that the method was linear up to a homocysteine concentration of 140  µmol/L, in comparison with HPLC of plasma homocysteine which showed a linearity up to 200  µmol/L. The coefficient of variation within a batch of measurement was 5% and within batches was 8%. HPLC of plasma samples was only 1% within batch and 4% between batches. Storage of the dried blood sample spots for more than 24 hours in room temperature decreased the homocysteine concentration, probably due to the hemolysis of whole blood during drying. The hemolysis could have been accompanied by the denaturation of enzymes that release homocysteine from blood cells. Overall, although the use of dried blood samples offers flexibility in sampling and storage, the assay had lower precision compared to HPLC. It may not be able to quantify dried blood homocysteine in newborns because its lower limit of detection is above the levels that some healthy neonates have. The method can be used in the detection of homocysteinuria. A more rapid protocol for tHcy measurement is liquid chromatography electrospray ionization-tandem mass spectrometry (LC/MS/MS) (Gempel, et al., 2000). It removes the derivatization step and can perform 400 analyses daily. Blood samples are applied to filter cards that are used for neonatal homocysteine screening. The cards are incubated in the presence of a reducing agent and a deuterated internal standard. The tHcy is then extracted in an acetonitrile solution and loaded on a cyano column. Eluates are ionized with an ion spray device before passing through a mass spectrometer. The procedure makes use of an autosampler, which hastens the analysis. The advantage of the method is the removal of the derivatization step, because the mass spectrum is determined directly. Impurities of the samples are removed by passing them through the cyano column. Since the procedure takes only 3 minutes to finish 1 sample, more samples can be processed in less time compared to HPLC (Gempel, et al., 2 000). An innovation in a high throughput liquid chromatography-tandem mass spectrometry method is the use of a 96-well plate format that skips precipitation, centrifugation and derivatization. Plasma samples, calibrator and control were mixed well in 96-well plates. From each well, an aliquot was transferred to another plate with sodium hydroxide solution (diluent) and dithioethritol (or another suitable reductant). After thorough shaking and a short incubation period, the plate was placed in the autosampler for LC-MS/MS analysis. Comparison of the results with those obtained by HPLC showed that the LC-MS/MS method gave consistently higher homocysteine readings, which was attributed to the use of different standards by the two methods. The biggest advantage of the new method over HPLC was the expense, because LC-MS/MS reduced the costs for material and manpower by more than 50% (Arndt, et al., 2004). Enzyme Immunoassay The immunoassay-based tests for homocysteine were developed in the 1990s. Most assays employ a mouse monoclonal antibody against S-adenosylhomocysteine (SAH), which is formed when adenosine and S-adenosylhomocysteine hydrolase are added to reduced total homocysteine of the plasma/serum sample. The earliest report on the procedure involved the use of flouresceinated SAH as tracer. This method is fluorescent polarization immunoassay or FPIA. After SAH is generated, it is mixed with a fluorescein-tagged SAH. The mixture is coated onto a microtiter plate, then made to bind with mouse anti-SAH (Figure 6). When antibody-SAH bind, the changes in fluorescent polarization of the SAH-tracer are detected and serve as basis for the quantification after a standard curve of known homocysteine calibrators is constructed (Shipchandler and Moore, 1995). The newer versions of enzyme immunoassays were performed on microtiter plates and used the formation of colored complexes to measure absorbance which was used as the basis of the quantification (Figure 7) (Frantzen, et al., 1998). After the conversion of reduced tHcy to SAH, another enzyme is added to the mixture to hydrolyze the excess adenosine. After reaction with a monoclonal anti-SAH antibody, a specific anti-mouse-antibody is added, which forms a colored product that can be detected spectrophotometrically for quantification. The immunoassays are simpler to perform compared to chromatography because they do away with the derivatization step and the expensive equipment needed in chromatographic separation. The method has been proven to be rapid and precise. The coefficient of variation between and within assay is 8% and less than 6% respectively, and the results also correlated well with those obtained using HPLC. The advantage of the immunoassay over HPLC is speed and ease of determination (Frantzen, et al., 1998). Many samples can be run simultaneously on several microtiter plates, which make the immunoassay ideal for routine screening. Furthermore, the technique can be and has been fully automated using robotics technology, and is highly flexible and can be used on different analytical platforms. Other similar imunoassays have been developed. In a variant of the assay, recombinant homocysteine lyase, converts homocysteine into ÃŽÂ ±-ketoglutarate, with the evolution of ammonia and H2S (Tan, et al., 2000). A second reaction reacts H2S with N,N-Dibutylphenylene diamine (DBPDA) resulting in the formation of form 3,7-bis(dibutyl amino)phenothiazine-5-ium chloride, a highly fluorescent compound. Both highly purified recombinant homocysteine lyase and DBPDA were synthesized in the authors laboratory, which somehow limits the utility of the procedure because their production requires skill and specialized equipment. Another assay utilizes the recombinant enzyme, methionine ÃŽÂ ³-lyase, and the commercially available fluorophore N,N-diethyl-p-phenylenediamine (Chan, et al., 2005). The rest of the procedure is similar to that used in typical immunoassays. The method can be used in an automated analyzer or manually, which makes it ideal for laboratories that cannot afford the expensive automated analyzers. Although the recombinant enzyme was purified only from a crude extract of E. coli, the results compared well with those that were obtained using HPLC. These results showed that the need for tedious and careful purification procedures can be removed. However, a main drawback was the non-specificity of the enzyme for homocysteine; it also reacts, although with less specificity, with cysteine which is present in larger amounts than homocysteine in the plasma. Thus, the use of pure homocysteine as calibrator is not suitable for the assay. Instead, the recommendation is to use pooled plasma as the calibrator, but this requires the determination of the tHcy by HPLC, which complicates the procedures for small laboratories who do not have access to HPLC equipment. Moreover, the precision of the procedure is also lowered if samples are diluted, because this also reduces the amount of cysteine in the sample relative to its concentration the calibrators. The presence of cysteine is also the cause for the lower sensitivity of this assay, because at lower tHcy levels the background fluorescence due to cysteine is amplified (Chan, et al., 2005). Enzyme cycling The most recent method for tHcy determination is the enzyme cycling method which amplifies the detection signal to improve the sensitivity of the detection (Figure 8) (Dou, et al., 2005). Similar to the previous methodologies, all protein-bound and oxidized forms of homocysteine are first reduced to free homocysteine. Next, methionine and S-adenosyl homocysteine (SAH) are formed from the methylation of free tHcy by the action of the enzyme homocysteine methyltransferase with S-adenosyl methionine as methyl donor or co-substrate. The SAH so formed comes from SAM and the transmethylation reaction, and not from the free homocysteine molecules. Upon formation, SAH is hydrolyzed immediately to homocysteine and adenosine by SAH hydrolase. The homocysteine originating from SAM enters the homocysteine transmethylation reaction catalyzed by homocysteine methyltransferase to form homocysteine and methionine again, resulting in an enzymatic cycle and an increase in detection signals. The reacti ons cause the accumulation of adenosine, which is converted to inosine and ammonia through the action of adenosine deaminase. NH3, in the presence of NADH and 2-oxoglutarate are converted by glutamate dehydrogenase to glutamate, water and NAD+ . The amount of tHcy is proportional to the concentration of NAD+ in solution, which is determined by its absorbance at 340 nm. The first commercially available HCY assay developed has three homogenous reagents that were developed for analyzers that can handle 3 reagents. If the analyzer can handle only 2 reagents, reagent 1 and 2 are mixed before the run (Diazyme, 2010). Only 20  µL or less of sample volume is necessary. The duration of the entire test is only 10 minutes whether 2 or 3 reagents are used. This method has been tested on many analytical analyzers of different brands. The method is highly sensitive, differences in homocysteine concentrations lower than 1.5  µmol/L can be detected. The assay can be used to measure homocysteine concentrations of 3 to 50  µmol/L, which is lower than that of other procedures. However, 50  µmol/L is beyond the upper limit of most normal adults. An innovation to the enzyme cycling method described previously utilizes the conversion of pyruvate to lactate and the amount of NAD+ produced (Roberts and Roberts, 2004). Bound and oxidized homocysteine is reduced, and reacted with serine to form cystathionine. Cystathionine ÃŽÂ ²-lyase is added to the mixture to catalyze the degradation of cystathionine to homocysteine, pyruvate and ammonia. Pyruvate is reduced to lactate by the action of lactate dehydrogenase, and the concomitant oxidation of NADH to NAD+. The amount of NAD+ produced is directly proportional to the amount of homocysteine in the sample, and can be detected spectrophotometrically at 340 nm. This method can detect up to 0.31 ÃŽÂ ¼mol/L of homocysteine. Compared to the earlier enzyme cycling technique, this method was linear from 1 to 100 ÃŽÂ ¼mol/L, meaning that it can detect homocysteine concentrations as high as 100  µmol/L albeit with a maximum deviation of T Polymorphism and Risk of Coronary Heart Disease: A Meta-analysis. JAMA, 288 (16), pp.2023-2031. Kluijtmans, L., Young, I., Boreham, C., Murray, L., McMaster, D., McNulty, H. et al. [2003] Genetic and nutritional factors contributing to hyperhomocysteinemia in young adults. Blood , 101, pp.2483-2488. Koehler, K., Baumgartner, R., Garry, P., Allen, R., Stabler, S. and Rimm, E. [2001] Association of folate intake and serum homocysteine in elderly persons according to vitamin supplementation and alcohol use. American Journal of Clinical Nutrition, 73, pp.628-637. 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Refsum, H., Grindflek, A., Ueland, P., Fredriksen, A., Meyer, K., Ulvik, A., et al [2004] Screening for serum total homocysteine in newborn children. Clinical Chemistry, 50, pp.10-25. Refsum, H., Smith, D., Ueland, P. N., Clarke, R., McPartlin, J., Johnston, C., et al. [2004] Facts and recommendations about total homocysteine determinations: an expert opinion. Clinical Chemistry, 50, pp.3-32. Riddell, L., Chisholm, A., Williams, S., and Mann, J. [2000] Dietary strategies for lowering homocysteine concentrations. American Journal of Clinical Nutrition, 71, pp.1448-1454. Roberts, R., and Roberts, W. [2004] Performance characteristics of a recombinant enzymatic cycling assay for quantification of total homocysteine in serum or plasma. Clinica Chimica Acta, 344 (1-2), pp.95-99. Selhub, J. [1999] Homocysteine metabolism. Annual Review of Nutrition, 19, pp.217-246. Selhub, J., and Miller, J. 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My Personal Philosophy Of Teaching Essay - 2027 Words

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