Immunology Coursework Example | Topics and Well Written Essays - 1000 words

Immunology - Coursework Example In 1 sentence or less, clarify what could clarify the disappointment of this immunization. 21. In 2002, Diane Mathis and associates found the capacity a particle named AIRE that was basic for the counteraction of multi-organ immune system illness. How might you best depict the capacity of AIRE? Immunization alludes to the infusion of a microorganism in order to quicken the safe framework to counter the organism, in this manner forestalling ailment. Inoculations or Vaccinations, work through incitement of the invulnerable framework. The basic concern, it is illogical to oversee cell procedures to ensure that an infection that is replication-lacking won't normally become replication-capable. In the event that an infection transforms into replication-equipped, it conceivably will re-procure any qualities of pathogenic associated with the wild-type germ and could prompt sickness. Lymphocytes are accepted to enter the lymph hubs through a course of steps. Stage 1: Cells at first move on and tie endothelium by means of selectin-intervened communications; at that point experience an invigorated improvement in integrin action (stage 2. It at that point triggers them to stand solidly by endothelium through the association of Ig super-family particles of bond and integrins (stage 3). The cells that are solidly connected at that point trail a chemoattractant slant into tissues (stage 4). The integrins and selectins fundamental for the initial step include L-selectin. Stage 3 requires lymphocyte integrin LFA-1 (aLb2) that intervenes firm attachment through obliging to its ICAM-1, HEV-communicated counter-receptors. The atoms that are proficient in performing stage 2 and stage 4 capacities are Chemokines. CD28 and CTLA-4 are homologs, which each predicament B7; be that as it may, have restricting jobs. Despite the fact that CD28 co-animates T cells, CTLA-4 forestalls enactment of T cells through blocking cell cycle movement and cytokine creation. CTLA-4 is quickly prompted after actuation of T cells and has astounding inhibitory properties.

The Art of Food free essay sample

One day on the transport following a hauling 8 hours of school I couldnt stand by to return home, lie in bed and watch some TV, which was my standard daily practice after school. At the point when I returned home that day I stuffed my jacket and book sack in the storage room and getting into some agreeable garments for my rest. At the point when I laid in bed I heard a thump on my entryway. â€Å"Ugh extraordinary what does my grandpa need now† I snorted. I shouted â€Å"come in!† and a head flew in the split of my entryway. He at that point inquired as to whether I needed to assist him with supper that he had a migraine and he didnt feel better. Obviously like some other youngster I fought no, gave him mentality and said I was worn out and needed to rest. He smoothly said â€Å" OK that’s fine.† After five minutes or so I was feeling regretful lastly got the vitality to get up and assist him with getting ready supper. We will compose a custom article test on The Art of Food or on the other hand any comparable theme explicitly for you Don't WasteYour Time Recruit WRITER Just 13.90/page He was making rice, chicken, and beans like he generally was nevertheless I never truly focused on how much work it truly took to make supper consistently for around 5 individuals. He showed me how to cook rice and cleave veggies and how to flame broil chicken. Everything was really interesting to me and I was somewhat having a fabulous time doing it. He at that point conversed with me about working in a kitchen in Cuba working from 7 am to 10 pm regularly so as to help my grandmother and their 5 children. He educated me regarding the woman who encouraged him all that he thought about cooking, Angelita Arias. She showed him how to truly acknowledge food and basically he grew an adoration for food and that’s what he was giving to me. Since that day I generally get back home from school anxious to help my grandpa in the kitchen and see what we were sauteing, searing, grilling, fricasseeing that day. My grandpa acquainted me with an actual existence I didnt know existed, somethin g people don’t truly focus on which is the craft of food, the specialty of transforming nothing into something supernatural and heavenly. Since the time that day I understood what I needed to do with my life, to turn into a culinary expert and offer my adoration for food with the world and I can just thank my grandpa for that.

Motion Creates Emotion 3 Simple Ways To Break Your Pattern

Motion Creates Emotion 3 Simple Ways To Break Your Pattern Its difficult to roll out of bed some days. Sometimes you just dont feel motivated: You dont want to create. You dont want to contribute. You dont want to do anything. Sometimes youd rather sit passively in front of the television and watch the world progress without you. We know, it happens to us, too. Sometimes we procrastinate or we simply dont feel like doing something. But then we do three things that allow us to break our pattern and take action. Good news, theyre all pretty easy: 1. Accountability Partners.  Together, your two authors,  exercise with each other five days a  week. It helps to have someone else there to make sure we show up when we need to show up and to make sure we give it a little extra effort each time. Who is your accountability partner? It can be via phone, in person, or even email or text. Get one. 2. Take the First Step. We tend to put things off, especially boring or mundane tasks (doing the dishes, laundry, writing, etc.), but if we take the first step, we get momentum and we are much more likely to complete the task. Sometimes we just need to take that first stepâ€"as difficult as that might seem at timesâ€"and the rest falls in line. 3. Change Your State. If we dont feel like getting up or doing anything, we will drastically change our physiology (go for a walk, do push-ups, change our breathing, run as fast as we can for 30 seconds, etc.). Motion creates emotion. These three things will get you moving in the right direction. Subscribe to The Minimalists via email.

The opportunities and pitfalls of cloud computing Free Essay Example, 2500 words

However, since the inception of the cloud computing, most of the barriers to the entry for starting a company have been dramatically reduced and the cloud computing may have removed this strategic advantage away from the large corporations (Cunningham, & Wilkins, 2009). This, therefore, leaves the small talented team of the individuals to address the perceived market needs. Additionally, this alleviates the need for the venture capital funds that provides the necessary IT infrastructures. Cloud computing creates a learning environment that can surpass the restrictions posed by the location and time that most students face. With the cloud computing, there is a learning environment whereby the students can log in to the online blackboards, downloading the course materials, taking the tests and exams or completing the assignments in a more flexible manner. Additionally, the cloud computing can increase the interoperability between the different institutions that are using the same clou d provider and hence promoting the joint research programs and the exchange of the vital scientific information and the databases between the researchers (Hwang, Dongarra, & Fox, 2013). The utilization of the cloud computing can lead to the reduction of the costs related to the IT infrastructure that are necessary for the standard internet network service. We will write a custom essay sample on The opportunities and pitfalls of cloud computing or any topic specifically for you Only $17.96 $11.86/pageorder now This, therefore, makes the cloud computing particularly very appealing to the governments and businesses that lack the financial means of covering the internet IT support and the recurring hardware costs. For instance, in the education sector, several universities, especially in the developing countries, use the Software as a Service (SaaS) applications whereby the cloud servers host the virtual interfaces of the educational contents. With the cloud services, the educational systems can facilitate the on-line courses, the payroll systems, student emails and the improvements of the cross-campus collaborations between the various university departments. Due to the flexible nature of the cloud services, the educational institutes and universities are able to use some of the cloud services on demand thus helping in the saving of the funds that can be allocated for other services. Another advantage of the cloud computing is in the health sector, especial ly in the developing countries whereby the majority of the people resides in the remote or rural areas. The mobile health applications are important for the health workers in delivering the efficient healthcare services to the rural areas by providing the range of the services.

Essay on Biography of George Washington the First American...

George Washington was the first president of the United States. I think as my opinion, George Washington being president was a major turning point in US history. Mr. Washington had an interesting early life, before he was the president. Same goes for his presidency and after his presidency. Here is a overall look at George Washington’s life. George Washington was born on February 11, 1732 in Westmoreland, Virginia, British America, in his parents Pope’s Creek Estate. George Washington was the first child of Augustine Washington and Augustine’s second wife, Mary Ball Washington. Augustine Washington belonged to the Colony of Virginias landed gentry and was a planter and slaveholder. Mary Bell Washington was a stay home mom just like most†¦show more content†¦After Lawrences death (1752), George inherited part of his estate and took over some of Lawrences duties as adjutant of the colony. As district adjutant, which made (Dec., 1752) him Major Washington at the age of 20, he was charged with training the militia in the quarter assigned him. George Washington married Martha Custis in 1759. Martha Custis was a rich widow who had two children named Martha and John, however George and Martha did not have children together (Wikipedia). Washington was unanimously elected President of the United States of America by electors in early 1789 and again in 1792. Both votes were unanimous. John Adams was his vice-president. Washingtons first inauguration took place in New York City, New York (which was the first capital of the USA, from 1789 to 1790). Washingtons second inauguration took place in Philadelphia, Pennsylvania (it was the capital from 1790 to 1800). Washington refused a third Presidential term, saying in his farewell speech that a longer rule would give one man too much power. George Washington remains the only president to have received 100 percent of the electoral votes. Washington was not a member of any political party and hoped that they would not be formed, fearing conflict that would undermine republicanism. During Washingtons presidency, the Bill of Rights was adoptedShow MoreRelatedAnalysis Of The Book Into The Wild By Jon Krakauer1686 Words   |  7 Pagesfreedom and exemption. Washington and McCandless are similar because they did something most people would not. They both stepped out of the ordinary society and decided to do what they think is best. For example, Washington led the Continental Army against the great British Empire, and Chris left his normal and traditional life, to seek a life of adventure and determined to go on a journey across the United States. Washington became the first president of the United States. Washington was also the onlyRead MoreHow Do Americans View George Washington and Christopher Columbus Today?772 Words   |  4 PagesWashington had an impact on America during the late 1700’s, and Columbus had an impact on what he hoped was the West Indies in the late 1400’s and early 1500’s. George Washington and Christopher Columbus are viewed in two complete different ways. Columbus found America and brought the early English settlements over. Washington helped found our country. However even though both men affected our country both are viewed differently than today. George Washington set a legacy that we are still followingRead MorePresident George Bush Essay894 Words   |  4 PagesGeorge W. Bush was born in New Haven, Connecticut on July 6, 1946. Just like other presidents, he had his good times and his bad times. He was the forty-third president of The United States. Because of those who had preceded him in The Oval Office, he understood the importance of being the Commander in Chief. In all of the events that happened during his presidency he never gave up on his country (Biography; Gale: Vol 21). The Bush family moved to Texas when George was only two years old. GeorgeRead MoreBipgraphy of George Washington Essays1015 Words   |  5 PagesYes, George Washington was our first president so he had to be a good leader, but everybody struggles. Of course, this did apply to George Washington and he was still the best man to begin our country’s government. To discover the details, however, investigation and explanation is required. Born in 1732 at Wakefield Plantation in Virginia, George Washington became his father’s eldest of six children in his second marriage. His father did his best to provide him with tutors and private schoolsRead MoreAccomplishments And Failures Of George Washington1738 Words   |  7 Pages21 April 2017 Many Triumphs and Failures of George Washington. George Washington had enormous amounts of failures, but he had achieved great things as well. From being a young boy living in Pope’s Creek, Virginia to becoming the president of the United States. He has faced a lot of hardships and failures. But through all of his hardships throughout his life he has become the greatest president of the United States, George Washington. George Washington had early influences as a child. His childhoodRead MoreAccomplishments And Failures Of George Washington1738 Words   |  7 Pages21 April 2017 Many Triumphs and Failures of George Washington. George Washington had enormous amounts of failures, but he had achieved great things as well. From being a young boy living in Pope’s Creek, Virginia to becoming the president of the United States. He has faced a lot of hardships and failures. But through all of his hardships throughout his life he has become the greatest president of the United States, George Washington. George Washington had early influences as a child. His childhoodRead MoreGeorge Washington : Man, Myth, Legend Essay1193 Words   |  5 PagesFrom cutting a cherry tree at the age of six to being the first president of the United States George Washington was a very influential man throughout his career. The topics of discussion will be his life before, during, and after his career, his major events, and the effects of his major events on the world. There shall be three subtopics amongst each main topic. For the first section, the subtopics are the importance of his birthdate and birthplace, childhood life, and his education. The secondRead MoreGeorge Washington : The American Of American National History Essay1288 Words   |  6 PagesGeorge Washington stands at the origins of American national history. Many Americans call him â€Å"the father of our country†. Washington commanded the Continental Army in the Revolutionary War. He led the American colonists to freedom fro m British rule. He headed the convention that created the U.S. Constitution, as president of the republic Washington has created a basis for a federal government in which the Americans were able to find their national agreement. Despite his accomplishments as a generalRead MoreAnalysis of William M Thayers A Biography of George Washington: The Patriot President691 Words   |  3 Pages In his biography of Americas founding father and first president, George Washington: The Patriot President, author William M. Thayer describes the heroic acts of leadership that Washington displayed while fighting in the American Revolution and during his two terms as President. Using a personal style that appeals to young readers, Thayer provides many glimpses into the private life of George Washington that give a better impression of the man behind one of historys most legendary figures. ByRead MoreAccomplishments And Failures Of George Washington1765 Words   |  8 PagesFailures of George Washington. As a young boy, George Washington grew up in a small town in Pope’s Creek, Virginia.   Ã‚  Ã‚  He came from this small town in hopes of achieving great things, like becoming the first president of the United States.   He faced many failures, but by overcoming them, he made many great achievements.   Although, he faced hardship and failures, he did not let those get in the way of his goals.   He became one of the greatest presidents of the United States of America. George Washington

Christopher Marlowe Works During The Renaissance Free Essays

Many major and influential authors emerged during the Renaissance. Among these talented individuals was Christopher Marlowe. Marlowe and his fellow writers of the late sixteenth and early seventeenth centuries, impacted the course of writing, which preceded their life. We will write a custom essay sample on Christopher Marlowe Works During The Renaissance or any similar topic only for you Order Now Their works continue to be read and studied by numerous people, to this day. Christopher Marlowe was a dominant English poet and playwright, who perhaps was William Shakespeare†s most important predecessor in England (Britannica 917). Of all writers in the Elizabethan era, he was perhaps the most dashing, tempestuous, and appealing (Microsoft Encarta). Although Marlowe was considered the most important dramatist, prior to Shakespeare, his entire career as a playwright lasted only six years. Marlowe was born on February 6th, 1564 in Canterbury, England. His father, John Marlowe, was a shoemaker and tanner. His mother, Catherine Author, was the daughter of a clergyman. Marlowe attended Kings School in Canterbury, England. At Kings School, he received a very regimented education, which was considered one of the best available during that time. The school day began and ended with a prayer at six am and five p. m. respectively. In addition to daily instruction in religion and music, they also sang the morning mass in the Cathedral. The boys were allowed to speak solely in Latin, even while at play. He was granted a scholarship, established by Matthew Perry, to attend Corpus Christi College in Cambridge. (Gale Research) After receiving his BA in 1584, he became known as â€Å"Dominus† Marlowe(. At age twenty-one, his motto was â€Å"That which nourishes me, destroys me† (Kunitz 823). This statement foretold and shaped his writing style. From thereafter, many absences from the university were recorded. In 1587, he was allowed to obtain his Masters, only after the Privy Council sent a letter to the university making it very clear that his service to the government had frequently taken him abroad. He left Cambridge after six and a half years of study with the intention of taking holy orders and entering the Anglican Church, as ordained by his scholarship. However, instead he entered the government service, as an agent. In edition, Marlowe became a playwright for the London theatres. As an occasional actor in 1589, Marlowe maintained his role as a regular dramatist for the Lord Admirals Company. He also wrote for The Earl of Nottinghams Companies. Marlowe was often in trouble with the law, and little is known about his life, aside from his writings. He led an adventurous and dissolute life. Earlier playwrights concentrated on comedy, while Marlowe worked on tragedy, and advanced it considerably as a dramatic medium. By uncovering the great possibilities for strength and variety of expression in blank verse, Marlowe helped to establish the verse as the predominant form in the English drama. His first successful play, Tamburlaine the Great, was divided into two sections, parts one and two. It appeared that originally Marlowe intended to only write the first part. However, the popularity of the first part motivated him to create a second part. It was produced at Rose Theatre from 1587 to 1588, and published by R. Jhones in 1590. The ambition-maddened hero, in Tamburlaine the Great, was a shepherd, who desired the crown, luxury and power. However, his conquests were damaged by his absurdity. Tamburlaine marked the birth of the Shakespearean drama. The following lines from Tamburlaine illustrate Marlowe†s opinion of human glory: Nature that fram†d us of four elements Warring within our breasts for regiment, Doth teach us all to have aspiring minds: Our souls, whose faculties can comprehend The wondrous architecture of the world, And measure every wandering planet†s course, Still climbing after knowledge infinite, And always moving as the restless spheres, Wills us to wear ourselves and never rest, That perfect bliss and sole felicity, The sweet fruitition of an earthly crown. (Britannica 917) His masterpiece, The Tragical History of Doctor Faustus, told of a man selling his soul for the price of all knowledge. This appealed to the superstitious Elizabethans. It was produced in 1594 as Dr. Faustus at the Rose Theatre. V. Simmes published The Tragical History of Doctor Faustus in 1604. The following lines from this work show the possible redemption through Christ†s blood: The stars move still, time runs, the clock will strike, The devil will come, and Faustus must be damn†d. O, I†ll leap up to my God! -Who pulls me down? – See, see, where Christ†s blood streams in the firmament! One drop would save my soul, half a drop: ah, my Christ! – Ah, rend not my heart for naming of my Christ! Yet will I call on him: O, spare me, Lucifer! – Where is it now? ’tis gone: and see, where God Stretcheth out his arm, and bends his ireful brows! Mountains and hills, come, come, and fall on me, And hide me from the heavy wrath of God! (Britannica 918) In addition to writing plays, Marlowe also wrote poetry. His major poetic work was Hero and Leander. This was incomplete at his death, the first two sections were completed at his death and the remaining two were finished by George Chapman in 1598, almost five years after Marlowe†s death. Marlowes writing here shows an amazing skill in the management of the narrative decasyllable couplet. No agreement was reached as to the composition date of this poem, but it is ranked as a major work along with Tamburlaine and Faustus. Marlowe began writing poetry and performing plays, when he entered Kings School. His education shaped him to become the genius who first created the Shakespearean blank verse drama. This is why many hailed him as â€Å"The Morning Star†, of the turning point from comedy to drama in England (Marlowe Society Home Page). Marlowe had a dangerous reputation for being â€Å"atheist†; however, he could have had just unorthodox beliefs. He was aquatinted with Sir Walter Raleigh, who was adventurous in his religious beliefs. Marlowe was summoned on May 18th, 1593, to appear before the Privy Council for accounts of â€Å"atheism and immortality†. The charge was a heresy and a most serious crime; the ultimate penalty was burning at the stake. Despite the seriousness of the crime, Marlowe was released bail, however he had to appear at the court daily. Marlowe escaped to Debtford, England. He stayed at Dame Eleanor Bull†s house who hired out rooms and served meals. Marlowe was murdered there on May 30th, 1593, at age 29. The strange circumstances for Marlowes murder in that room in Debtford, have been the subject for many debates. Four men were present at the house on that day Robert Poley was an experienced government agent who carried the Queens most secret letters to and from the courts in Europe. He had arrived from Debtford, straight from The Hague, where he had been on the Queens business. Igram Frizer was the personal servant and business agent of Marlowe†s patron, Thomas Walsingham. Nicholas Skeres often assisted Poley. Poley, Skeres and Frizer were all experienced con men and liars. Also present that day was Christopher Marlowe. Some believe that the cause of his murder was a dispute over a dinner bill. Others believed that Marlowe was murdered because the three men believed that he knew too much about the government. Although the facts surrounding his death are unknown, the end result was Marlowe†s death by stab wounds. Igram Frizer was accused of the murder of Marlowe, however he was pardoned on June 28th, 1593. Marlowe was buried in Debtford on June 1st, 1593. The death of Christopher Marlowe led many scholars to theorize that he faked his death and assumed the name William Shakespeare to escape the Privy Council. Shakespeare was born two months after Marlowe, and he became very popular shortly after the death of Marlowe. Little is known about Shakespeare other then mentioned of his poor education. There is not much evidence of his existence other then baptism paper, and a will left leaving his bed to his wife. There is no mention of his works or manuscripts in his will Marlowe began his career as a poet and playwright towards the end of the Renaissance. The Renaissance was the period from about 1350 to 1600 in which European scholars revived the learning of ancient Greece and Rome. It was a period in European history that saw a renewed interest in the arts. In this time, the middle ages and feudal times were transformed into a society dominated by the arts. It was a time of achievements in the arts and sciences as well as a period when people were deeply concerned with religious issues. Renaissance is a French word meaning â€Å"rebirth†. Scholars reacted against what they saw as the â€Å"dark ages† of medieval Europe and revived the learning of ancient Greece and Rome. Like painting and sculpture, literature expressed the attitudes of the Renaissance. The middle class formed a demanding new audience, which enjoyed dramatic tales rather the comedies. Literature was often written in the common language, but some continued to write in Latin. Literature emphasized religious as well as worldly themes. Invention of printing during the Renaissance greatly increased the number of book available. In the fourteenth century, Europeans learned from Arabs who had previously learned from the Chinese how to make paper from rags and wood pulp. The fifteenth century invention of movable type by Johann Gutenberg also shaped the way literature had previously been written. Gutenberg developed a type of metal that could be used to make movable type. In 1455, the first complete edition of the bible was printed; this started the era of printed books. More and more pieces began to be printed up, and thus decreased the cost of books, making them more available and more popular (Beers 245). Each of Marlowe†s play included a protagonist with single passion that dominated them. The character is doomed to destruction due to their desire of power. He had a background of classical and theological learning. Marlowe â€Å"turned blank verse into a supple instrument for dramatic expression†. (Gale Research). Marlowe worked on tragedy and advanced it immensely. â€Å"Marlowe shows the power to view a tragic hero from more then one angle, achieving a simultaneous vision of grandeur and impotence† (Microsoft Encarta). Renaissance impacted Marlowe due to the desire of the middle class for tragedy rather then comedy, which had previously been the focus of European literature. Marlowe was one of the first writers of drama and tragedy that shaped the history of Europe. Marlowe laid the foundation of tragedy of what would rise up as a new era, known as the Shakespearean Era. Writing was a competition and was taken very seriously by writers including Marlowe. Writers are impacted tremendously by the time period that is taken place during their careers. The Renaissance was a time of new ways of writing and expressing ideas, which impacted the attitudes of the people. The writers changed their style to reflect the time and attitudes. They influence the minds of people to think about their ideas and opinions on a particular subject. Writers can persuade people concerning a topic by revealing the positive and negative aspects of it. Marlowe was an influential and powerful writer. It was unfortunate that his career was cut short. He had a very promising career, but his own decision to become a government agent endangered his career and stopped him from becoming an more popular and productive playwright. How to cite Christopher Marlowe Works During The Renaissance, Essay examples

System Studies and Technological Advances

Question: Discuss about the System Studies and Technological Advances. Answer: Introduction: In this experiment ischemia can be triggered by the mediators released from the tissues. Receptors specific to the cardiomyocytes like A1 and A3, bradykinin B2, 1-adrenergic, AT1 angiotensin II, muscarinic M2, and -opioid receptors can be stimulated by the released mediators. Both single and multiple pathway ischemia protection can be achieved by using freshly isolated cardiomyocytes. Isolated cardiomyocytes experiment would be valuable tool for the evaluation of the mechanistic insight of the ischemia. In comparison with the whole heart experiment, use of cardiomyocytes would be helpful in eliminating endothelial cells and fibroblasts, which would affect analysis. Pure cardiomyocytes would give more robust analysis of desired cell signaling as compared to the whole heart. Visibility using microscope is the most prominent advantage of cardiomyocytes. More number of cells can be made available in freshly isolated cardiomyocytes as compared to the cell cultures. Limitations: Isolated cardiomyocytes cannot be used for longer time and repeated use of cardiomyocytes is not possible. In the isolation of cardiomyocytes from blood vessel and extracellular matrix, there is the possibility of loss of vital features of pathophysiology of ischemia. Isolated cardiomyocytes are not capable of reproducing ischemia mechanisms similar to the whole heart 1 . Methods: Cardiomyocytes will be isolated form isolated heart by enzymatic digestion using calcium-free medium containing collagenase and trypsin. Isolated cardiomyocytes will be centrifuged at very slow speed to get pellet with fluid spread over pellet with fluid volume of one-third volume of the pellet. Supernatant will be covered with oil and will be incubated at 37C. Preconditioning will be performed by incubation in the absence of glucose. Evaluation of occurrence of ischemia and effect of Bob on the ischemia in cardiomyocytes will be evaluated using Tryptan Blue staining. Four set of isolated cardiomyocytes will be there like normal cardiomyocytes, ischemia induced cardiomyocytes, ischemia induced cardiomyocytes incubated with different concentrations of Bob and cardiomyocytes incubated with Cyclosporin A. Cyclosporin A will be used as positive control in this experiment. Cyclosporin A was already evaluated in the cardiomyocytes ischemia experiment and it was proved that Cyclosporin A exhibited its action through cytochrome C which is the mechanism of action Bob. Negative control will be used without any treatment and it will useful in the evaluation of induction of the ischemia in the isolated cardiomyocytes. Following are the end results of this experiment. Viability of the cells using Tryptan Blue will be useful in the evaluating recovery of cardiomyocytes from ischemia after treatment with Bob. Autofluorescence of NADH in the mitochondrial matrix will be evaluated to assess the redox state of the cardiomyocytes. This redox state will be helpful in inferring the severity of ischemia in negative control, Bob treated cardiomyocytes and Cyclosporin A treated cardiomyocytes. Langendoorf rat heart perfusion set up to allow ischemia-reperfusion Objective: To evaluate effect of Bob on myocardial ischemia in Langendoorf rat heart perfusion. Rationale: Langendorff isolated heart is a robust model for evaluating ischemia reperfusion injury. Advantages: Langendoorf rat heart perfusion set up can be used to evaluate real-time pharmacological treatments. It is also useful to get the uniform size of the infracted tissue and also useful in the application of drug directly to the myocardium. In vivo models are useful only in the end point analysis ischemia reperfusion injury, while Langendoorf rat heart perfusion is also useful in the functional analysis of the ischemia reperfusion in real time. Limitations: Isolated heart is continuously deteriorating and it can be useful only for short duration. Isolated heart preparations are denerveted and due to this exact pathological ischemia are difficult to achieve. Methods: Heart will be isolated. Aorta will be cannulated to the reservoir containing oxygenated perfusion fluid. Perfusion fluid will be contained in the sintered glass jacketed reservoir above the isolated heart. Regional ischemia will be induced by perfusing heart with KrebsHenseleit solution. Perfusion will be continued for 15 minutes and ischemia will be induced for 30 minutes. Apoptotic cells will be identified by the dUTP nick end labelling (TUNEL) using CardioTACS. Heart tissue will be fixed in the 3.7 % formaldehyde for 24 hours and will be embedded in the paraffin. Mitochondria will be isolated and mitochondrial respiration will be measured using Clarke-type oxygen electrotode. Cyclosporin A will be used in the washing buffer. Skinned cardiac fibers will be prepared and these fibers will be used for measurement of oxygen uptake. Level of cytochrome c will be measured by solubilizing cardiac fibers in Triton X-100. Cytochrome c will be measured using western blotting. Protei ns will be identified using primary monoclonal antibodies to cytochrome c using ECL western blotting detection reagents. Caspase activity in the heart homogenate will be measured in sucrose based medium. Cytosolic protein will be incubated with buffer and z-DEVD-pnitroanilide which is a caspase 3substrate. Caspase will be measured using spectrophotometer at 405 nm. Cyclosporin A will be used as positive control in this experiment. Heart will be preperfused with Cyclosporine A for about 15 minutes. Cyclosporine A protects heart from ischemia by preventing loss of cytochrome c. Use of Cyclosporine A is valid in this experiment because, Bob also exhibits similar mechanism of action. In this experiment, negative control will be without any type of treatment. This negative control will be useful in the evaluation of induction of ischemia and also evaluation of treatment effect of Bob and cyclosporine A. Results: Cyclosporine A and Bob will be reducing level of cytochrome c release from the ischemic isolated rat heart. Pretreatment of heart with Cyclosporine A and Bob will be resulting in the no decrease in the respiratory rate of mitochondria which will be measured using membrane potential. Also, dependence of cytochrome release on caspase will be evaluated in this experiment. Cyclospprine A and Bob will be inhibiting ischemia-induced caspase activation. Transient coronary artery ligation Objective: To evaluate effect of Bob on transient coronary artery ligation model of ischemia-reperfusion. Rationale: Transient coronary artery ligation model exhibits exact ischemia pathology. Advantages: Availability of experimental rat makes this model more feasible. Possibility of genetic modification in rat leads to the development of more specific model. Possibility of reperfusion makes this model make more clinically relevant because reperfusion always occurs in clinical setting prior to treatment. Ischemic period can be effectively controlled in this model based on the requirement of the intervention. Limitations: Variability of infract size is more in this rat surgical model. Variation of the infract size is mainly depends on the location of ligation, hence it is very important to select same location for all the animals in the study. Range of infract size is between 20 90 % of the entire left ventricle and as a result this model requires large number of animals in a study. Variability due to modification in the surgical procedure has also been reported from different laboratories. Also, there is reported variability due to different time of analysis. Despite the benefits of reperfusion, there is the possibility of damage of part of the myocardium 8-9. Methods: Rats will be approved from the Institutional Ethical Committee. Rats will be housed and procedure will be carried out in the aseptic condition. Rats will be anesthetized with suitable anesthetic agent. Chest and neck region of the rat will be shaved and will be placed in supine position under a stereomicroscope. Transient coronary artery ligation will be performed by making incision in the skin, chest muscle will be separated and thoracotomy will be performed in the fourth intercostals space. Pericardium will be removed and left coronary artery will ligated at 1 mm distal end of the tip of left auricle with the help of 7-0 polypropylene suture. Ligated artery will be released after 30 minutes of ischemia. Rats survived after surgery will be assigned to different treatment groups like curaglutide and Bob. Sham control group will performed with the similar surgical procedure without ligation of the coronary artery. Drugs will be administered to the animals intraperitoneally at different time points after reperfusion like 0, 5, 15 and 30 minutes. Sham control animals will be treated with normal saline. An additional set of animals will be utilized for the evaluation of caspase 3 activity. Each sample will be homogenized with RIPA buffer containing protease inhibitor and will be centrifuged. Fluorescence of the protein in treatment groups will be measured and will be compared with protein of the sham treated group to evaluate activity of caspase 3 activity. Tunnel assay will be performed to measure apoptotic rate. After the antigen retrieval it will be incubated with monoclonal antibody against actin and it will be stained with 6-diamidino-2-phenylinidole (DAPI) 10-12. Curaglutide will be used as positive control as it already proved its efficacy in the transient coronary artery ligation model. Negative control will be considered as the group with surgery without any type of treatment. Results obtained will be decrease in the caspase 3 activity and reduction in the infract size. Other hemodynamic parameters will also be evaluated like left ventricular systolic pressure and heart rate. References: Diaz, RJ, and Wilson GJ. Studying ischemic preconditioning in isolated cardiomyocyte models. Cardiovascular Research, vol. 70, no. 2, 2006, pp. 286-96. Lolita, Portal, Valrie, Martin, and Rana, Assaly. 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