Risk Management in Childrens Play

Hazard Management in Childrens Play Adjusted Approach to Risk Management A hazard is a piece of regular day to day existence and it nearly influences everybody in some structure or the other. A hazard is taken each day so as to support a superior, solid future. In this way chance can't be totally wiped out in any part of our general public thus tolerating that the chance of even genuine or hazardous wounds can't be wiped out, nonetheless, it ought to be legitimately and legally oversaw. Hazard ought to be recognized and controlled at a reasonable level to profit the potential open doors we receive consequently. A decent way to deal with overseeing hazard is basic to structure our general public. The idea of hazard/advantage appraisal ought to be a manual for accomplish fulfillment. Recess is basic and indispensable piece of a childs life. Recess legitimately influences a childs prosperity and advancement. The Health Safety Executive (UK) recognizes that No youngster will learn on the off chance that they are enclosed by cotton fleece (http://www.hse.gov.uk/diversion/childrens-play-july-2012.pdf) Through recess youngsters create various aptitudes, for example, fearlessness, group building, and social abilities in school. One of the key advantages of hazard is; offspring of different age bunches find out about hazard themselves, generally through open air play exercises. A fair way to deal with overseeing hazard for giving childrens play openings and exercises just as the advantages they increase out of it ought to be painstakingly estimated. Perceiving the presentation of hazard may shape some portion of play openings. This improves their experience to assume liability for their own wellbeing and prosperity as they grow up. The National Travel Survey 2014 has affirmed that the most widely recognized method of transport to class by kids between the ages of 5-10 is by strolling or vehicle. 88% of kids between the ages of 7-10 were joined by a grown-up and the greater part of them concurred that street security was their greatest worry; among different concerns. While, a hazard is a piece of life yet mishaps don't need to be; by and large mishaps can be effectively maintained a strategic distance from. A recent report by, The English Outdoor Council concedes that On an average school visit, the youngsters who partake are at no more serious danger of death than their classmates who have remained behind. http://www.englishoutdoorcouncil.org/wp-content/transfers/Nothing-Ventured.pdf So as to protect youngsters, kids must be instructed deliberately; person on foot aptitudes; street crossing, safe cycling and hurrying from an early age and ought to be connected overwhelmingly between the ages of 7-10. Most grade schools have straightforwardly connected these exercises to their educational program. These abilities will assume an imperative job in overseeing hazard for the yr 6 Residential Visits, Outdoor School Activities, The advantages picked up from these exercises ought to be adjusted against the peril or hazard it might have. In my school, we have an Adventure play area which is utilized by all students. Be that as it may, KS1 is just permitted to utilize the Adventure play area every so often as Marble rewards for 15mins. KS2 utilizes the Adventure play area on rota premise each play time. The greatest kids permitted to get to the Adventure play area is roughly 30. Understudies are not permitted to get to the play area on wet days, indoor play exercises are sorted out. At the point when understudies utilize the Adventure play area, the class instructor actually screens the play territory. A far reaching site study is done each term to oversee hazard. All kids figure out how to utilize the experience offices in an alternate manner. They all need to appreciate and play and arrive at explicit targets. KS1 utilize the offices and evaluate every action at their own pace, while KS2 particularly the 10/multi year old are more grounded and arrive at their particular targets and objectives at a quicker pace. The experience they gain in the Adventure play area, with no grown-ups meddling increase them strength and capacities as they figure out how to deal with their own hazard. Kids with a particular incapacity would be observed by a manager on a 1:1. Another movement in our school is Climb the Wall accessible just to yr 5/6. The action itself is testing and it assembles students certainty; profiting them with climbing development, readiness, parity and coordination, attention to bodys focus gravity. A hazard is surveyed and overseen by fitting utilization of head protectors, footwear, garments, long hairs tied back, no adornments, and pockets all discharged. The movement is carefully directed by 1:1 proportion; 3 kids permitted at a solitary time. Kids with a particular incapacity would not be permitted to take part in this action. One of the action our school composes is a Residential excursion just to year 6. Kids experience different advantages, some of them are building individual certainty, group building action, and creating social abilities and investigating their qualities and shortcoming. The targets and the result of this movement are very much overseen and estimated in the outing. The hazard is overseen by the proportion of understudies to the instructor. This action isn't accessible to some other understudies in the school as the targets and advantages are thin. Our school knows about the wellbeing and danger to kids. The school has custom fitted a selective Road Safety Education into their educational plan. Year 5 youngsters are offered cycle preparing. This program is encouraged by the East Riding Council and a gathering of 4/5 youngsters are taken outside the school with their cycles and caps and prepared to utilize a cycle securely. The hazard is evaluated and overseen by the school just as the ERC. Kids who pass this preparation are permitted to cycle to class autonomously. A passerby ability program is masterminded yr 4, by and by composed by the ERC and Scoot the Route is offered to year 3. Kids in Yr 6 experience a Bus Aware program which will profit them before they go to the auxiliary school. Reference NA. (2012). CHILDRENS PLAY AND LEISURE PROMOTING A BALANCED APPROACH . Accessible: http://http://www.hse.gov.uk/diversion/childrens-play-july-2012.pdf . Last got to 12/02/14. Jill T. (NA). Nothing Ventured. Available:http://www.englishoutdoorcouncil.org/wp-content/transfers/Nothing-Ventured.pdf Last got to 14/2/17.

Free Essays on Southeast Asia

Southeast Asia Have you at any point been strolling through your lawn and all the unexpected your leg was brushed off by a landmine? Well 1 in each 236 individuals in Cambodia has had that occur. That’s why numerous individuals have fled their homes in Southeast Asia since they don’t need to put them selves or their family at risk for getting exploded by a landmine. Numerous individuals have started to return back to their homes after numerous years from them, since they feel that they can manage the landmines and that possibly some of mines are no more. There are more than 40 million evacuees on the planet that have left their homes on account of the danger of death or injury from landmines. â€Å"One study†¦ shows that during a multi month time frame in 1992, 85 percent of those that where harmed in mine blasts were occupied with nonmilitary exercises, for example, cultivating, going among towns, and simply tending their cattle.† So there are a great deal of guiltless individuals getting injured to no end. Most mines are found essentially in the creating scene along streets, ways, and fields. This is causing numerous individuals the option to have a decent free life, on the grounds that a significant number of the individuals don’t need to take a shot at ranches or heard dairy cattle since they never know when they may step on a mine. This makes them go poor and afterward they can’t bolster their family. In Cambodia, one woman was out allowing her cows to cattle and as she was strolling around taking care of her steers she happened to step on a mine. This harmed both of her legs, making her family offer all their dairy cattle to take care of off the medical clinic tab and now they have no cash. Landmines are principally utilized in wars for exploding fighters; this puts a dread among the adversary. They are additionally used to secure things that are situated on the ground, similar to bases, land rockets, or even limits. Yet, when the war is over what befalls the land mines that have not been exploded? All things considered, they simply sit and sit, holding on to carry out their responsibility, and t... Free Essays on Southeast Asia Free Essays on Southeast Asia Southeast Asia Have you at any point been strolling through your lawn and all the unexpected your leg was passed over by a landmine? Well 1 in each 236 individuals in Cambodia has had that occur. That’s why numerous individuals have fled their homes in Southeast Asia since they don’t need to put them selves or their family at risk for getting exploded by a landmine. Numerous individuals have started to return back to their homes after numerous years from them, since they believe that they can manage the landmines and that possibly some of mines are no more. There are more than 40 million displaced people on the planet that have left their homes in view of the danger of death or injury from landmines. â€Å"One study†¦ shows that during a multi month time span in 1992, 85 percent of those that where harmed in mine blasts were occupied with nonmilitary exercises, for example, cultivating, going among towns, and simply tending their cattle.† So there are a great deal of guiltless individuals getting injured in vain. Most mines are found principally in the creating scene along streets, ways, and fields. This is causing numerous individuals the option to have a decent free life, in light of the fact that a large number of the individuals don’t need to take a shot at ranches or heard steers since they never know when they may step on a mine. This makes them go poor and afterward they can’t bolster their family. In Cambodia, one woman was out allowing her dairy cattle to cattle and as she was strolling around taking care of her cows she happened to step on a mine. This harmed both of her legs, making her family offer all their steers to take care of off the emergency clinic tab and now they have no cash. Landmines are essentially utilized in wars for exploding officers; this puts a dread among the foe. They are likewise used to secure things that are situated on the ground, similar to bases, land rockets, or even limits. Yet, when the war is over what befalls the land mines that have not been exploded? All things considered, they simply sit and sit, standing by to carry out their responsibility, and t...

The Sneaky Role of Some Additives in Cigarettes

The Sneaky Role of Some Additives in Cigarettes Addiction Nicotine Use The Inside of Cigarettes Print The Sneaky Reasons Behind Some Cigarette Additives By Terry Martin facebook twitter Terry Martin quit smoking after 26 years and is now an advocate for those seeking freedom from nicotine addiction. Learn about our editorial policy Terry Martin Medically reviewed by Medically reviewed by Sanja Jelic, MD on June 08, 2016 Sanja Jelic, MD, is board-certified in sleep medicine, critical care medicine, pulmonary disease, and internal medicine.   Learn about our Medical Review Board Sanja Jelic, MD Updated on February 14, 2020 Design Pics/Kelly Redinger/Getty Images More in Addiction Nicotine Use The Inside of Cigarettes After You Quit How to Quit Smoking Nicotine Withdrawal Smoking-Related Diseases Alcohol Use Addictive Behaviors Drug Use Coping and Recovery As of Dec. 20, 2019, the new legal age limit is 21 years old for purchasing cigarettes, cigars, or any other tobacco products in the U.S. In April of 1994, a master list of 599 additives used in the production of American cigarettes was made public, with five major tobacco companies reporting. Up to that time, the constituents of cigarettes were unknown to anyone but the companies producing them.  Today, while many questions still remain, researchers have uncovered plenty of useful information about the additives in cigarettes, and it all started with that list. For the most part, cigarette manufacturers maintain that their additives add flavor, act as a humectant or are used as a processing aid, as in this list of additives from Philip Morris.  Science shows us that much more is going on, however. Researchers Michael Rabinoff, DO, PhD, Nicholas Caskey, PhD, Anthony Rissling, MA, and Candice Park, BS reviewed publicly available tobacco industry documents, the list of 599 additives and other sources. The findings:  More than 100 of them have qualities  that may either enhance addiction or mask the negative effects of cigarettes. Whether the additional properties of tobacco additives were designed into commercial cigarettes by Big Tobacco is unproven, but researchers all agree that strict regulatory control over tobacco additives is of vital importance. The Effects of Cigarette Additives Dilate the airways, allowing the smoker to inhale more deeply and deposit higher levels of tar in the lungs. Cocoa is an additive known to do this.Addictive potential of their own or synergistically with nicotine.Slow the metabolism of nicotine, increasing smokers exposure.Anesthetic properties that decrease the harshness of tobacco smoke on the throat.Mask smell, visibility, and irritation of environmental tobacco smoke.Disguise warning symptoms of illnesses associated with cigarette smoking. Common Tobacco Additives Tobacco documents show that cigarette companies have investigated ways to develop cigarettes that will hook the user more quickly and thoroughly to nicotine,  researching everything from enhancing the nicotine in cigarettes through genetically engineered tobacco plants to developing nicotine extracts and using sheet tobacco as an additive in the manufacturing process.?? A Long List of Cigarette Ingredients Ammonia Ammonia added to cigarettes reacts with nicotine in a process call free-basing.  The result is a bigger nicotine kick for the smoker. What Does Ammonia in Cigarettes Do? Menthol Derived from mint oils, menthol is another additive thought to play a significant role in favorably introducing young people to cigarettes.  Menthol is a mild local anesthetic, and when added to cigarettes can ease throat irritation caused by cigarette smoke.?? Eugenol is another additive that is used as a numbing agent for the harsh qualities of cigarette smoke. Are Clove Cigarettes a Healthy Way to Smoke? Acetaldehyde Acetaldehyde is formed when cigarette ingredients and additives, including sugars are burned.  Animal research conducted by Philip Morris showed a synergistic effect between acetaldehyde and nicotine. Rats pressed a bar more for the combination of the two chemicals than for either one alone. If the data were generalized to humans, it would relate to increased puffs on a cigarette due to the synergistic effect of nicotine and acetaldehyde in tobacco smoke. Sheet Tobacco Cigarette manufacturers reconstitute bits of leftover tobacco plant stems, stalks, scraps, collected dust, and floor sweepings into an ingredient that can be added back into the final cigarette product. The process involves grinding all of the above materials, extracting the nicotine from them, and then adding glue, fillers, chemicals, and other agents to form a slurry.  The mixture is then pressed into a sheet, puffed up, and the extracted nicotine is sprayed on it.  From there, it is ground up once again into fine curls that are incorporated into cigarettes in the desired amount. Sheet tobacco is a major ingredient in modern manufactured cigarettes. Toxic Byproducts of Burning Cigarettes As mentioned above, the burning of benign food additives by themselves and in combination with other additives can create new chemical compounds that are hazardous to human health.?? And, to make matters worse, pesticides used in tobacco farming and heavy metals found naturally in the soil, some of which are radioactive,  can linger and travel on to the finished product (and the consumer).   Get Help to Quit Smoking Addiction tells us there is no good time to quit, so ignore the urge to put it off and start your quit program today. Quit Smoking and Make It Stick!

Kant s Categorical Imperative Essay - 1454 Words

In this paper, I will explain the concept of Kant’s Categorical Imperative, and show how he used it to justify why it is wrong to lie to an inquiring murderer. I will note how he arrived at this conclusion, and why I consider it to be the correct moral answer. According to Kant, the Categorical Imperative is the supreme law of morality by which a particular rule that an individual takes as a maxim must be accepted by all rational beings. This universal acceptance is what judges an action to be always good, provided that the agent’s impartiality and independence are maintained over self interest. Therefore, the Categorical Imperative is the only valid criterion by which to decide whether an action is permissible. It is an unconditional and absolute obligation (duty), which even desire (inclination) cannot override. Duty derives from reason, which only humans possess, and thus they are the only beings capable of judging right from wrong. Kant’s Categorical Imperative is a pure a priori form, that is, one that is based on theoretical as opposed to empirical deduction. It is a rational, voluntary choice derived from finality, instead of causality, where interests are put aside and moral duty is enforced. Take, for example, the challenge Benjamin Constant presented to the Categorical Imperative in the journal France for 1797, Part VI, No. 1, pages 123 124, in which he affirmed that Kant would not lie to a murderer if the latter asks you whether your friend, whom he isShow MoreRelatedKant s Categorical Imperative Essay1246 Words   |  5 Pagesutilitarianism, Kant was more focused on intent and action itself. This leads into one of Kantian ethics main ideals; you mustn’t treat another human being as a means to an end. Kant’s Categorical Imperative (CI) is a deontological theory, which relied heavily on his belief that humans are all capable of reason in the same manner, on the same level (A Brief Summary of Kant s Categorical Imperative, 2012). Kant recognized 2 kinds of moral ‘imperatives’, a hypothetical imperative (what must be doneRead MoreKant s Categorical Imperative Essay1190 Words   |  5 PagesIn section I of Immanuel Kant’s categorical im perative, Kant argues that every human being alive is subject to the categorical imperative. Kant came to this conclusion by arguing that the only thing that is good without needing qualification is a good will. Throughout this paper I will discuss Kant’s good will and his three propositions. A good will is an act done from duty and motivated by respect. If a person manifests a good will in action, the respect for duty determines that I do the actionRead MoreKant s Categorical Imperative Essay2239 Words   |  9 PagesKant’s Categorical Imperative An imperative is the linguistic form of a ‘command of reason’. In section II of the Fundamental Principles of the Metaphysics of Morals, German philosopher Immanuel Kant writes, â€Å"the conception of an objective principle, in so far as it is obligatory for a will, is called a command (of reason), and the formula of the command is called an imperative.† It is a rule telling us what we ought to do. He distinguishes between two types of imperatives: hypothetical and categoricalRead MoreKant s Categorical Imperative Essay1448 Words   |  6 PagesKant’s categorical imperative is a natural conclusion of reason when searching for a moral guideline that does not depend on previous expense but reason alone. The categorical imperative can be explained in many different ways. Kant offers five formulations in his work groundwork of the metaphysics of morals. The formulations of Kant’s categorical imperative can be considered a test. If your maxim passes th e test then your actions under that maxim will be good. The formulations that Kant offers,Read MoreKant s Categorical Imperative Essay983 Words   |  4 PagesImmanuel Kant is known for his absolute and idealistic approach to answering this question, with which he provides us a medium to answer it. Kant calls this his categorical imperative. Throughout this paper I will break down Kant’s view on ethics, explain one formulation of his categorical imperative, and evaluate his theory on an existential level. Kant was a firm believer that there are two different worlds. He called them the â€Å"World of Phenomena,† and the â€Å"World of the Noumena.† Kant describesRead MoreEssay about Kant ´s Theory: Categorical Imperative and its Rules827 Words   |  4 Pageswill benefit them at the end. The purpose of Kantianism is to tell us that morality is not to make us happy but the whole purpose is to do the right thing just for the sake of doing it. Eventually doing the right thing will lead us to our happiness. Kant said that we are determined to know what’s good or what’s bad through self- law and using ourselves as our own guidance’s. We as individuals will determine our own behavior than having someone telling us how we should act because of their expectationsRead MoreKant And The Moral Law1451 Words   |  6 PagesIntroduction: Kant argues that mere conformity with the moral law is not sufficient for moral goodness. I will argue that Kant is right. In this essay I will explain why Kant distinguishes between conforming with the moral law and acting for the sake of the moral law, and what that distinction means to Kant, before arguing why Kant was right. 2) Meaning of Kant’s Statement Why: According to Kant, we can control the will and meaning behind our actions. The morality of an action should be assessedRead MoreKant s Philosophy On Moral Philosophy Essay1519 Words   |  7 PagesAlthough Kant s philosophy, outlined in The Groundwork for the Metaphysics of Morals, has some value as a moral guide, it alone is not always sufficient. After analysing Kant s objective moral imperative, I will show that implementing his philosophy has the potential to permit what is considered to be immoral. This is due to its rigid conditions, which are formed on the basis that he believes in the existence of a perfect morality that always holds true. However, I will argue that there is notRead MoreKant And Mill On Animal Ethics Essay1365 Words   |  6 PagesIn this essay I will begin by explaining the overall views of Immanuel Kant and John Stuart Mill, then compare and contrast the ideas and philosophies of Kant and Mill on Animal Ethics. I believe that Kant, the deontologist, will not care as much about the duty/responsibility between humans and animals as Mill, the utilitarian, who will see the extreme importance of animal ethics. After studying and explaining the views and teachings of these two philosophers I will see if my thesis was correct,Read MoreThe Ones Who Walked Away from Omelas by Ursula Le Guin1424 Words   |  6 Pagesgreatest number of people. On the contrary, Kant would argue that using the child as a mere means is wrong and argue that the living conditions of the child are not universalizable. The citizens of Omelas must face this moral dilemma for all of their lives or instead choose to silently escape the city altogether. My central thesis is that Kant would give the child’s life inherent value and advocate that Omelas’ citizens abandon their practices. In this essay I aim to examine the story of Omelas through

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How Are Civil Liberties Better Protected Since 1997 Free Essays

To what extent have civil liberties become better protected since 1997 In comparison to countries such as France and the USA, the UK’s commitment to civil liberties was weak. During the US declaration of civil Independence the US made amendments to its constitution,; their bill of rights became largely made up of civil liberties, including the right to freedom of religion, speech etc. And the French Revolution led to the establishment of the Declaration of the Rights of Man and the Citizen (1789). We will write a custom essay sample on How Are Civil Liberties Better Protected Since 1997 or any similar topic only for you Order Now Whereas, the UK has traditionally been reluctant to give basic rights and freedoms explicit legal expression. Instead, it relied on the freedoms that were supposed to be embodied in the common Law belief that ‘everything is permitted that is not prohibited’. However, in recent years, the protection of civil liberties has increasingly fallen to the courts, due to the wider use of the power of judicial review and the introduction of the Human Rights Act. One of the roles of Judiciary is to ‘Defend Civil liberties’ though Judicial review, As one of the Human rights Act is Habeus corpus- the right to a fair trial. Judges can overrule government if they are going beyond Ulta-Vires, judges can decide that other political actors are acting beyond their proper power, in recent years, judges have been increasingly willing to use this power, particularly in relation to ministers, for example if police arrest you without given reason of arrest, they are acting beyond ultra-vires, because you have a right to be given a reason for arrest and can therefore take them to court. However, judges cannot overturn acts of Parliament because of Parliamentary sovereignty unlike in the USA, judges have very far-reaching powers of judicial review because of the existence of a codified constitution. If a law passed by congress goes against the constitution or the bill of rights – the Supreme Court can overrule that law. The Human Rights Act is a UK law passed in 1998. It means that you can defend your rights in the UK courts and that public organisations (including the Government, the Police and local councils) must treat everyone equally, with fairness, dignity and respect. ——————————————– [ 1 ]. Andrew Heywood – Essentials Of Politics- ’Protecting civil Liberties’ Pp. 287 How to cite How Are Civil Liberties Better Protected Since 1997, Essay examples

The aim of this research is, eventually, to Essay Example

The aim of this research is, eventually, to Essay Context of the intended research CRIT is a receptor that was foremost encountered on the surface of aSchistosomaspecies ; in theSchistosoma,it acts as a decoy C2-binding receptor in order to protect this parasite from complement onslaught by viing with C4 for the binding of C2 ( Inal and Schifferli, 2002 ) . Complement is, basically, an enzyme system that is triggered upon immune system onslaught: most of the enzymes in this system are identified through standardised labeling: they are labelled C followed by a figure and so a codification based on the cleavage merchandises when proteolysed, for illustration, C5b-9. Complement onslaught ( Carroll, 1998 ) plays a major function in supporting hosts from immune onslaught, in footings of extinguishing foreign encroachers, and involves a complex tract of interactions, in order that the procedure does non take to self devastation: so, unregulated complement action can take to autoimmune diseases ( Ohet al. ,2003 ) , and other conditions/diseases such as bosom onslaught ( K ilgoreet al. ,1994 ) , Alzheimer’s disease ( Bradtet al. ,1998 ) . We will write a custom essay sample on The aim of this research is, eventually, to specifically for you for only $16.38 $13.9/page Order now We will write a custom essay sample on The aim of this research is, eventually, to specifically for you FOR ONLY $16.38 $13.9/page Hire Writer We will write a custom essay sample on The aim of this research is, eventually, to specifically for you FOR ONLY $16.38 $13.9/page Hire Writer CRIT ( or Sh-TOR as it was antecedently known ) was found inSchistosoma,as a complement C2 protein, that could barricade complement activation, therefore bring oning bilharzia in worlds ; it was hypothesized that the CRIT blocked complement activation through its C2 binding site ; CRIT look in the parasite therefore acts as a steerer C2 binding receptor, protecting the parasite against complement onslaught by viing with C4 for the binding of C2, at the ed1 sphere ( Hui, 2005 ) . CRIT has later been found that other animate beings, that are suited as theoretical account systems for analyzing CRIT action besides express CRIT, for illustration, the rat andTrypanosoma.Recent work has besides shown that worlds have a CRIT factor, labeled ( Hu ) -CRIT which is expressed in a broad scope of human cells, particularly in hematopoietic cells ( Inalet al. ,2005 ) . Further elaborate surveies of CRIT have shown that it is a transmembrane receptor with two extracellular and two in intracellular spheres with an active 11 amino-acid peptide subdivision ( called CRIT-H17 ) which is hypothesized to be involved in the complement suppression activity of CRIT ( Hui, 2005 ) . Much work has been undertaken on clarifying the mechanism of action of CRIT, in footings of it being a potentially utile molecule in handling immunological diseases and other diseases, such as malignant neoplastic disease. For illustration, Inalet Al.( 2005 ) have shown that CRIT barricading can take to deprotection in CRIT-expressing human myeloid cell lines and in monocytes, ensuing in a greater susceptibleness to complement-mediated lysisin vitro. Other research, such as that by Mollet Al.( 2006 ) has shown that CRIT look is different in patients with kidney upsets, through assorted alterations in up- or down-regulation of CRIT look, taking to the suggestion that the upregulation of CRIT in activated podocytes might stand for a self-defense mechanism, stand foring a ‘last line’ of defense mechanism in membranous kidney disease of the kidney. Other surveies have looked at the existent mechanism of operation of the CRIT tract, for illustration Huiet Al.( 2005 ) looked at the look of a functional recombinant von Willebrand factor-A sphere from human complement C2, in footings of this being a possible binding site for C4 and CRIT. As we have seen, CRIT competes with C4b for the binding of C2, with the major adhering site on C2 being located on a short peptide sequence that was antecedently of unknown beginning ; Huiet al. ,( 2005 ) , nevertheless, looked at a part on C2 that was known to hold binding capacity, the von Willebrand Factor-A, and found that, so, this peptide sequence inhibits complement activity ; utilizing a cloned von Willebrand Factor-A sequence, Huiet Al.( 2005 ) were able to look in item at the interactions between C2 and CRIT and C4b. This pilot survey that is intended to be undertaken as portion of this reappraisal of CRIT is intended to take this work of Huiet Al.( 2005 ) farther. As will be seen, on adh ering the serum complement protein, C2, the CRIT peptide, H17 can cut down complement-mediated redness in vivo [ 7 ] and it is intended that CRIT-ed1 and H17 will be tested as possible distinction therapeutics peculiarly aiming monocytic leukemia. To better the efficaciousness of these peptides, structural information on the manner they interact with C2, such that this can be ‘tweaked’ to increase efficiency of binding. The CRIT-H17 peptide will be synthesized full length as an 11-mer, but besides as a 10-mer, 9-mer and 8-mer. We will besides mutate to alanine the amino acids believed to interact with the vWFA sphere of C2. CRIT-H17 will farther be synthesized as a head-to-toe cyclised version of H17 ( to mime the native CRIT molecule ) . Prior to the structural surveies, nevertheless, these peptides will be tested for their efficaciousness at cut downing complement activationin vitro. In add-on, interactions of these peptides with C2 vWFA sphere will be monitored for by a novel technique utilizing electrospray ionization mass spectroscopy ( developed within the Institute for Health Research A ; Policy at LMU by Dr. A. Bligh ) [ 8 ] . This technique can besides observe the presence of two adhering sites and if there are two ligands whether they bind competitively and with what affinity. To supervise conformational alterations on interaction, in add-on to working out dynamicss of interaction, we will utilize Double Polarisation Interferometry ( AnaLight Quantum ) through coaction with Dr. R.B. Sim ( MRC Immunochemistry Unit, Univ. of Oxford ) . Prof. Peter Gros ( Centre for Biomolecular Research, Utrecht ) has approached the applier for CRIT peptides for co-crystallisation with an available C2a ( von Willebrand Factor A [ vWFA ] and serine peptidase ) concept and we expect this coaction to continue and to finally demo the points of contact between CRIT-ed1 and C2 ( via the vWFA sphere ) . It is hoped that this methodological analysis will let a deeper apprehension of how CRIT binds with other molecules in the complement tract, and how this tract is regulated in footings of supplying a intervention option for some of the diseases/conditions that are known to develop following perturbation, or irregular operation of, this complement tract. Introduction The purpose of this research is, finally, to transport out a pilot survey to back up preliminary grounds that the complement receptor CRIT ( Complement C2 Receptor Inhibitor Trispanning ) plays a function in monocyte/macrophage terminal distinction. We aim to demo that by triping distinction through CRIT, it is possible to suppress the proliferation of myeloid leukemia cells. This could hold of import deductions for an alternate intervention agenda for monocytic leukemia. By the terminal of this survey we expect to demo the undermentioned: I ) that monocytes which can adhere C2 through CRIT maintain monocytic phenotypein vitroorin vivo ;II ) that monocytes can be induced to terminally distinguish by barricading the interaction of CRIT with its blood relation ligand, complement C2 or as monocytes move into an extravascular environment devoid of C2 ; that a knockdown of CRIT look in monocyte carcinoma cell lines or physical blocking of CRIT-C2 interaction induces them to terminally dis tinguish, and that, likewise, a blocking of CRIT-C2 interaction on leukaemic monocytes will halt proliferation and bring on distinction ; and, in conclusion, that, conversely, monocytes that are C2-/- , can non be induced to terminally distinguish by barricading the interaction of CRIT with its ligand, C2, nevertheless on traveling into an extravascular environment, other excess mechanisms, such as interaction of monocyte integrins with VCAM molecules on endothelial cells, may still bring on distinction. In consequence, basically, either barricading ligand interaction with CRIT or cut downing CRIT look should excite cell rhythm apprehension ( irreversible block at G1 ) and terminal distinction into cells with a macrophage phenotype. These conditions represent a fresh tract for monocyte/macrophage terminal distinction, based on the theoretical account proposed, and affecting the complement receptor, CRIT. In footings of monocyte leukemia, besides advancing cell rhythm apprehension, barricading CRIT with anti-CRIT-ed1 has the added benefit of rendering the cell more susceptible to complement-mediated cytolysis, as described antecedently for monocyte carcinoma cell lines ( U937 and THP-1 ) and primary monocytes showing CRIT [ 1 ] . It is intended that this research will lend to a deeper apprehension of how the complement tract works in worlds, in peculiar with respect to how abnormalities in the operation of the complement tract can do disease, and how CRIT look modulates the operation of the complement tract in human systems. The informations obtained from this pilot survey will be used to look into five chief issues: I ) The function of CRIT in myeloid distinction The function of CRIT in myeloid distinction has been studied with an antagonist CRIT-based peptide termed H17 ( NH2-HEVKIKHFSPY-CO2H ) consisting the 11aa C-terminus of CRIT-ed1. Preliminary work suggests that in adhering to C2 and therefore barricading the interaction of C2 with CRIT [ 1 ] , H17 may bring on the distinction of monocytes/promonocytic cell lines along the tract of macrophage distinction, and significantly, inhibit cell proliferation. The different curative attacks that are presently used in handling acute leukemia include cytotoxicity, programmed cell death and distinction. Differentiation therapy was developed over a decennary ago and Acts of the Apostless by bring oning cell rhythm apprehension and hence distinction in leukaemic monocytes [ 2 ] , therefore elegantly avoiding cytotoxicity effects. Retinoids, such as all-trans-retinoic acid ( ATRA ) are used to handle promyelocytic leukemia by specifically aiming neoplastic cells whilst non impacting normal mature ce lls. Many successes in the intervention of monocytic leukaemias have been reported since [ 3-5 ] . We suggest that a CRIT-based peptide ( H17 or an H17 derived function ) could finally offer an of import alternate intervention for monocytic leukemia by bring oning distinction of monocytic cells. The peptide will be tested entirely and in combination with ATRA in a mouse theoretical account of acute promyelocytic leukemia [ 6 ] . two ) The construction of CRIT peptides ( ed1 and H17 ) and of CRIT peptides interacting with the von Willebrand Factor A ( vWFA ) sphere of complement C2 On adhering the serum complement protein, C2, the CRIT peptide, H17 can cut down complement-mediated redness in vivo [ 7 ] and we will prove CRIT-ed1 and H17 as possible distinction therapeutics peculiarly aiming monocytic leukemia. To better the efficaciousness of these peptides, we aim to utilize structural information on the manner they interact with C2. The CRIT-H17 peptide will be synthesized full length as an 11-mer, but besides as a 10-mer, 9-mer and 8-mer. We will besides mutate to alanine the amino acids believed to interact with the vWFA sphere of C2. CRIT-H17 will farther be synthesized as a head-to-toe cyclised version of H17 ( to mime the native CRIT molecule ) . Prior to structural surveies, these peptides will be tested for their efficaciousness at cut downing complement activation in vitro. Interactions of these peptides with C2 vWFA sphere will be monitored for by a novel technique utilizing electrospray ionization mass spectroscopy ( developed within the Institute for Health Research A ; Policy at LMU by Dr. A. Bligh ) [ 8 ] . This technique can besides observe the presence of two adhering sites and if there are two ligands whether they bind competitively and with what affinity. To supervise conformational alterations on interaction, in add-on to working out dynamicss of interaction, we will utilize Double Polarisation Interferometry ( AnaLight Quantum ) through coaction with Dr. R.B. Sim ( MRC Immunochemistry Unit, Univ. of Oxford ) . Prof. Peter Gros ( Centre for Biomolecular Research, Utrecht ) has approached the applier for CRIT peptides for co-crystallisation with an available C2a ( von Willebrand Factor A [ vWFA ] and serine peptidase ) concept and we expect this coaction to continue and to finally demo the points of contact between CRIT-ed1 and C2 ( via the vWFA sphere ) . three ) Expression profile of CRIT in autoimmune disease and malignant neoplastic disease With a position to understanding the function of CRIT in autoimmune disease and malignant neoplastic disease, the applier is join forcesing on a Swiss National Foundation funded undertaking with Prof. J. Schifferli ( Univ. Hospital Basel ) to do a CRIT smasher mouse. To happen an association between CRIT look degrees and the disease procedure, a comparing of CRIT look ( messenger RNA and protein ) in normal tissue with that in autoimmune disease and malignant neoplastic disease will be made. In situ hybridization surveies every bit good as immunohistochemistry, utilizing dual staining and/or staining of consecutive subdivisions with anti-CRIT and cell specific markers, will be conducted to corroborate look in sertoli cells, podocyte cells, keratinocytes, encephalon [ 1 ] . The distribution of CRIT in normal and morbid tissue will be studied, peculiarly tissues injured by inflammatory or necrotic procedures: joints-synovium in arthritis, myocardial infarction etc. Previously, we carri ed out a survey along these lines, which looked at CRIT look in assorted kidney diseases [ 9 ] . This survey revealed CRIT upregulation in membranous kidney diseases on glomerular podocyte cells. Unlike CR1, hitherto the lone other complement regulator described on podocytes, and which is non upregulated in membranous nephropathy, we believe that CRIT on podocytes represents a last line of defense mechanism against onslaught by complement. Functional information back uping this was later obtained ( manuscript in Complement regulators are frequently upregulated in malignant neoplastic disease [ 10 ] and so expression degrees of CRIT in assorted human malignances will be assessed excessively. Critically, CRIT is upregulated in liver malignant neoplastic disease ( Fig. 4 ) and thyroid malignant neoplastic disease ( non shown ) . As obstruction of CRIT with antibody sensitises cells to complement lysis [ 1 ] , such findings may hold applications in malignant neoplastic disease. Already schemes to barricade complement regulators with specific antibodies have been used successfully with a position to developing fresh malignant neoplastic disease immunotherapies [ 11,12 ] . In a recent development, membrane-bound complement regulative proteins ( mCRP ) have been downregulated by siRNA to render tumour cells sensitive to complement [ 13 ] . Therefore we will bring forth vector-based shRNAs ( utilizing psiRNA vector [ In vivo Gen ] ) to stably strike hard down CRIT look and see the consequence on tumor cells. four ) CRIT extracellular peptide ( H17 ) and its usage in modulating complement-mediated redness in in vivo theoretical accounts of autoimmune disease With a position to therapeutically suppress redness due to classical tract activation in theoretical accounts of complement-mediated autoimmune disease, CRIT has been targeted to suppress the Reversed Passive Arthus Reaction in mice [ 7 ] . By disposal of an counter peptide, H17, which binds complement C2 and prevents its activation it was possible to significantly cut down complement-mediated redness. To prove this peptide as a curative against autoimmune diseases in which the classical tract is peculiarly of import we are join forcesing with labs that have the appropriate animate being theoretical accounts. Experimental autoimmune myasthenia gravis ( EAMG ) is an antibody-mediated autoimmune disease impacting the neuromuscular junction. The disease, which is besides T cell-dependent, is an accurate theoretical account in footings of its pathology and clinical result of human myasthenia gravis ( MG ) . We have been approached by Prof. P. Christadoss and Dr. E. Tuzun of the Universit y of Texas, Galveston to prove H17 in their mouse theoretical account of MG [ 14,15 ] and will continue with this coaction. 1.2.5 Does CRIT adhere other serum proteins through its extracellular spheres? In a collaborative survey with Prof. Marina Botto ( Imperial College ) we will look into whether CRIT is a receptor for any other proteins beside complement C2 and FB, with which it binds with high and low affinity, severally. To prove whether the extracellular spheres of CRIT have other adhering spouses, receptor affinity chromatography [ 16 ] will be used to see whether ed1 binds other proteins from the serum of a C2 deficient ( C2D ) patient or of a combined C2/FB smasher mouse [ 17,18 ] . As the function of the 2nd extracellular sphere, ed2 has non been established, normal serum will be used ab initio to place ed2-binding proteins adhering to ed2 affinity columns by standard mass-spec designation protocols. Further experiments will be conducted to see if CRIT binds integrins. The principle for this is that CRIT-ed1 binds the vWFA1 sphere of complement C2 [ 19 ] and vWFA spheres are typically found in integrins, such as Very Late Antigen 4 ( VLA-4 ) on monocytes. The interaction w ith vascular cell adhesion molecule-1 ( VCAM-1 ) molecules on the endothelium is believed to non merely intercede attachment [ 20 ] and transendothelial migration [ 21 ] but besides to excite distinction [ 22 ] . Methodology to be utilized in the survey of the purposes of the undertaking: I ) The function of CRIT in myeloid distinction As we have seen, the function of CRIT in myeloid distinction has been studied with an antagonist CRIT-based peptide termed H17 ( NH2-HEVKIKHFSPY-CO2H ) consisting the 11aa C-terminus of CRIT-ed1. Preliminary work suggests that in adhering to C2 and therefore barricading the interaction of C2 with CRIT [ 1 ] , H17 may bring on the distinction of monocytes/promonocytic cell lines along the tract of macrophage distinction, and significantly, inhibit cell proliferation. It is intended that this pilot survey will go on the work that has been started in this respect, and will lend original research findings to the intervention of diseases that are caused by failures in the proper operation of the complement tract in worlds. The different curative attacks that are presently used in handling acute leukemia include cytotoxicity, programmed cell death and distinction. Differentiation therapy was developed over a decennary ago and Acts of the Apostless by bring oning cell rhythm apprehension and hence distinction in leukaemic monocytes [ 2 ] , therefore elegantly avoiding cytotoxicity effects. Retinoids, such as all-trans-retinoic acid ( ATRA ) are used to handle promyelocytic leukemia by specifically aiming neoplastic cells whilst non impacting normal mature cells. Many successes in the intervention of monocytic leukaemias have been reported since [ 3-5 ] . We suggest that a CRIT-based peptide ( H17 or an H17 derived function ) could finally offer an of import alternate intervention for monocytic leukemia by bring oning distinction of monocytic cells. The peptide will be tested entirely and in combination with ATRA in a mouse theoretical account of acute promyelocytic leukemia [ 6 ] . This will take to cons equences which could be of great usage in developing alternate therapies for handling conditions that arise as a consequence of failure of the right operation of the complement tract in worlds. two ) The construction of CRIT peptides ( ed1 and H17 ) and of CRIT peptides interacting with the von Willebrand Factor A ( vWFA ) sphere of complement C2 As has been seen, Huiet Al.( 2005 ) looked at the look of a functional recombinant von Willebrand factor-A sphere from human complement C2, in footings of this being a possible binding site for C4 and CRIT. As we have seen, CRIT competes with C4b for the binding of C2, with the major adhering site on C2 being located on a short peptide sequence that was antecedently of unknown beginning ; Huiet al. ,( 2005 ) , nevertheless, looked at a part on C2 that was known to hold binding capacity, the von Willebrand Factor-A, and found that, so, this peptide sequence inhibits complement activity ; utilizing a cloned von Willebrand Factor-A sequence, Huiet Al.( 2005 ) were able to look in item at the interactions between C2 and CRIT and C4b. The current survey will take the work of Huiet Al.( 2005 ) further, by looking in item at the CRIT tract, in footings of adhering the serum complement protein, C2, the CRIT peptide, H17 can cut down complement-mediated rednessin vivo[ 7 ] and we will prove CRIT-ed1 and H17 as possible distinction therapeutics peculiarly aiming monocytic leukemia. To better the efficaciousness of these peptides, we aim to utilize structural information on the manner they interact with C2. The CRIT-H17 peptide will be synthesized full length as an 11-mer, but besides as a 10-mer, 9-mer and 8-mer. We will besides mutate to alanine the amino acids believed to interact with the vWFA sphere of C2. CRIT-H17 will farther be synthesized as a head-to-toe cyclised version of H17 ( to mime the native CRIT molecule ) . Prior to structural surveies, these peptides will be tested for their efficaciousness at cut downing complement activation in vitro. Interactions of these peptides with C2 vWFA sphere will be moni tored for by a novel technique utilizing electrospray ionization mass spectroscopy ( developed within the Institute for Health Research A ; Policy at LMU by Dr. A. Bligh ) [ 8 ] . This technique can besides observe the presence of two adhering sites and if there are two ligands whether they bind competitively and with what affinity. To supervise conformational alterations on interaction, in add-on to working out dynamicss of interaction, we will utilize Double Polarisation Interferometry ( AnaLight Quantum ) through coaction with Dr. R.B. Sim ( MRC Immunochemistry Unit, Univ. of Oxford ) . Prof. Peter Gros ( Centre for Biomolecular Research, Utrecht ) has approached the applier for CRIT peptides for co-crystallisation with an available C2a ( von Willebrand Factor A [ vWFA ] and serine peptidase ) concept and we expect this coaction to continue and to finally demo the points of contact between CRIT-ed1 and C2 ( via the vWFA sphere ) . three ) Expression profile of CRIT in autoimmune disease and malignant neoplastic disease With a position to understanding the function of CRIT in autoimmune disease and malignant neoplastic disease, the applier is join forcesing on a Swiss National Foundation funded undertaking with Prof. J. Schifferli ( Univ. Hospital Basel ) to do a CRIT smasher mouse. To happen an association between CRIT look degrees and the disease procedure, a comparing of CRIT look ( messenger RNA and protein ) in normal tissue with that in autoimmune disease and malignant neoplastic disease will be made. In situ hybridization surveies every bit good as immunohistochemistry, utilizing dual staining and/or staining of consecutive subdivisions with anti-CRIT and cell specific markers, will be conducted to corroborate look in sertoli cells, podocyte cells, keratinocytes, encephalon [ 1 ] . The distribution of CRIT in normal and morbid tissue will be studied, peculiarly tissues injured by inflammatory or necrotic procedures: joints-synovium in arthritis, myocardial infarction etc. Previously, we carri ed out a survey along these lines, which looked at CRIT look in assorted kidney diseases [ 9 ] . This survey revealed CRIT upregulation in membranous kidney diseases on glomerular podocyte cells. Unlike CR1, hitherto the lone other complement regulator described on podocytes, and which is non upregulated in membranous nephropathy, we believe that CRIT on podocytes represents a last line of defense mechanism against onslaught by complement. Functional information back uping this was later obtained ( manuscript in Complement regulators are frequently upregulated in malignant neoplastic disease [ 10 ] and so expression degrees of CRIT in assorted human malignances will be assessed excessively. Critically, CRIT is upregulated in liver malignant neoplastic disease ( Fig. 4 ) and thyroid malignant neoplastic disease ( non shown ) . As obstruction of CRIT with antibody sensitises cells to complement lysis [ 1 ] , such findings may hold applications in malignant neoplastic disease. Already schemes to barricade complement regulators with specific antibodies have been used successfully with a position to developing fresh malignant neoplastic disease immunotherapies [ 11,12 ] . In a recent development, membrane-bound complement regulative proteins ( mCRP ) have been downregulated by siRNA to render tumour cells sensitive to complement [ 13 ] . Therefore we will bring forth vector-based shRNAs ( utilizing psiRNA vector [ In vivo Gen ] ) to stably strike hard down CRIT look and see the consequence on tumor cells. four ) CRIT extracellular peptide ( H17 ) and its usage in modulating complement-mediated redness in in vivo theoretical accounts of autoimmune disease With a position to therapeutically suppress redness due to classical tract activation in theoretical accounts of complement-mediated autoimmune disease, CRIT has been targeted to suppress the Reversed Passive Arthus Reaction in mice [ 7 ] . By disposal of an counter peptide, H17, which binds complement C2 and prevents its activation it was possible to significantly cut down complement-mediated redness. To prove this peptide as a curative against autoimmune diseases in which the classical tract is peculiarly of import we are join forcesing with labs that have the appropriate animate being theoretical accounts. Experimental autoimmune myasthenia gravis ( EAMG ) is an antibody-mediated autoimmune disease impacting the neuromuscular junction. The disease, which is besides T cell-dependent, is an accurate theoretical account in footings of its pathology and clinical result of human myasthenia gravis ( MG ) . We have been approached by Prof. P. Christadoss and Dr. E. Tuzun of the Universit y of Texas, Galveston to prove H17 in their mouse theoretical account of MG [ 14,15 ] and will continue with this coaction. 1.2.5 Does CRIT adhere other serum proteins through its extracellular spheres? In a collaborative survey with Prof. Marina Botto ( Imperial College ) we will look into whether CRIT is a receptor for any other proteins beside complement C2 and FB, with which it binds with high and low affinity, severally. To prove whether the extracellular spheres of CRIT have other adhering spouses, receptor affinity chromatography [ 16 ] will be used to see whether ed1 binds other proteins from the serum of a C2 deficient ( C2D ) patient or of a combined C2/FB smasher mouse [ 17,18 ] . As the function of the 2nd extracellular sphere, ed2 has non been established, normal serum will be used ab initio to place ed2-binding proteins adhering to ed2 affinity columns by standard mass-spec designation protocols. Further experiments will be conducted to see if CRIT binds integrins. The principle for this is that CRIT-ed1 binds the vWFA1 sphere of complement C2 [ 19 ] and vWFA spheres are typically found in integrins, such as Very Late Antigen 4 ( VLA-4 ) on monocytes. The interaction w ith vascular cell adhesion molecule-1 ( VCAM-1 ) molecules on the endothelium is believed to non merely intercede attachment [ 20 ] and transendothelial migration [ 21 ] but besides to excite distinction [ 22 ] . CRIT is a receptor that was foremost encountered on the surface of aSchistosomaspecies ; in theSchistosoma,it acts as a decoy C2-binding receptor in order to protect this parasite from complement onslaught by viing with C4 for the binding of C2 ( Inal and Schifferli, 2002 ) . Complement is, basically, an enzyme system that is triggered upon immune system onslaught: most of the enzymes in this system are identified through standardised labeling: they are labelled C followed by a figure and so a codification based on the cleavage merchandises when proteolysed, for illustration, C5b-9. Complement onslaught ( Carroll, 1998 ) plays a major function in supporting hosts from immune onslaught, in footings of extinguishing foreign encroachers, and involves a complex tract of interactions, in order that the procedure does non take to self devastation: so, unregulated complement action can take to autoimmune diseases ( Ohet al. ,2003 ) , and other conditions/diseases such as bosom onslaught ( K ilgoreet al. ,1994 ) , Alzheimer’s disease ( Bradtet al. ,1998 ) . CRIT ( or Sh-TOR as it was antecedently known ) was found inSchistosoma,as a complement C2 protein, that could barricade complement activation, therefore bring oning bilharzia in worlds ; it was hypothesized that the CRIT blocked complement activation through its C2 binding site ; CRIT look in the parasite therefore acts as a steerer C2 binding receptor, protecting the parasite against complement onslaught by viing with C4 for the binding of C2, at the ed1 sphere ( Hui, 2005 ) . CRIT has later been found that other animate beings, that are suited as theoretical account systems for analyzing CRIT action besides express CRIT, for illustration, the rat andTrypanosoma.Recent work has besides shown that worlds have a CRIT factor, labeled ( Hu ) -CRIT which is expressed in a broad scope of human cells, particularly in hematopoietic cells ( Inalet al. ,2005 ) . Further elaborate surveies of CRIT have shown that it is a transmembrane receptor with two extracellular and two in intracellular spheres with an active 11 amino-acid peptide subdivision ( called CRIT-H17 ) which is hypothesized to be involved in the complement suppression activity of CRIT ( Hui, 2005 ) . Much work has been undertaken on clarifying the mechanism of action of CRIT, in footings of it being a potentially utile molecule in handling immunological diseases and other diseases, such as malignant neoplastic disease. For illustration, Inalet Al.( 2005 ) have shown that CRIT barricading can take to deprotection in CRIT-expressing human myeloid cell lines and in monocytes, ensuing in a greater susceptibleness to complement-mediated lysisin vitro. Other research, such as that by Mollet Al.( 2006 ) has shown that CRIT look is different in patients with kidney upsets, through assorted alterations in up- or down-regulation of CRIT look, taking to the suggestion that the upregulation of CRIT in activated podocytes might stand for a self-defense mechanism, stand foring a ‘last line’ of defense mechanism in membranous kidney disease of the kidney. Introduction The purpose of this research is, finally, to transport out a pilot survey to back up preliminary grounds that the complement receptor CRIT ( Complement C2 Receptor Inhibitor Trispanning ) plays a function in monocyte/macrophage terminal distinction. We aim to demo that by triping distinction through CRIT, it is possible to suppress the proliferation of myeloid leukemia cells. This could hold of import deductions for an alternate intervention agenda for monocytic leukemia. By the terminal of this survey we expect to demo the undermentioned: I ) that monocytes which can adhere C2 through CRIT maintain monocytic phenotypein vitroorin vivo ;II ) that monocytes can be induced to terminally distinguish by barricading the interaction of CRIT with its blood relation ligand, complement C2 or as monocytes move into an extravascular environment devoid of C2 ; that a knockdown of CRIT look in monocyte carcinoma cell lines or physical blocking of CRIT-C2 interaction induces them to terminally dis tinguish, and that, likewise, a blocking of CRIT-C2 interaction on leukaemic monocytes will halt proliferation and bring on distinction ; and, in conclusion, that, conversely, monocytes that are C2-/- , can non be induced to terminally distinguish by barricading the interaction of CRIT with its ligand, C2, nevertheless on traveling into an extravascular environment, other excess mechanisms, such as interaction of monocyte integrins with VCAM molecules on endothelial cells, may still bring on distinction. In consequence, basically, either barricading ligand interaction with CRIT or cut downing CRIT look should excite cell rhythm apprehension ( irreversible block at G1 ) and terminal distinction into cells with a macrophage phenotype. These conditions represent a fresh tract for monocyte/macrophage terminal distinction, based on the theoretical account proposed, and affecting the complement receptor, CRIT. In footings of monocyte leukemia, besides advancing cell rhythm apprehension, barricading CRIT with anti-CRIT-ed1 has the added benefit of rendering the cell more susceptible to complement-mediated cytolysis, as described antecedently for monocyte carcinoma cell lines ( U937 and THP-1 ) and primary monocytes showing CRIT [ 1 ] . Bradt, B.M.et al. ,1998. Complement-dependent proinflammatory belongingss of the Alzheimer’s disease B-peptide.J. Exp. Med.188, pp.431. Carroll, M.C. , 1998. The function of complement and complement receptors in initiation and ordinance of unsusceptibility.Ann Rev Immun16,pp.545-568. Hui, K-M. , 2005. Biochemical and functional surveies of a fresh complement inhibitor, CRIT, with its interaction spouses. Dissertation submitted to the University of Basel, 2005. Hui, K-M.et al. ,2005. Expression of functional recombinant von Willebrand factor-A sphere from human complement C2: a possible binding site for C4 and CRIT.Biochem J.389, pp.863-868. Inal, J.M. , 2005. Complement C2 receptor inhibitor trispanning: from adult male to schistosome.Springer Seminars in Immunology27 ( 3 ), pp.320-331. Inal, J.M.et al. ,2005. Complement C2 receptor inhibitor trispanning: a fresh human complement inhibitory receptor.Journal of Immunology74, pp.356-366. Inal, J.M. and Schifferli, J.A. , 2002. Complement C2 receptor inhibitor trispanning and the B-chain of C4 portion a binding site for complement C2.Journal of Immunology168, pp.5213-5221. Kilgore, K.S.et al. ,1994. The complement system in myocardial ischaemia/reperfusion hurt.Cardiovasc Research28, pp.437. Makrides, S.C.et al. ,1992. Cell surface look of the C3b/C4b receptor ( CR1 ) protects the hamster ovary cells from lysis by human complement.J.Biol.Chem.34, pp.24754-24761. Moll, S.et al. ,2006. CRIT is expressed on podocytes in normal human kidney and upregulated in membranous kidney disease.Kidney International69,pp.1961-1968. Oh, K-S.et al. ,2003. Inhibition of complement activation by recombinant Sh-CRIT-ed1 parallels.Immunology110, pp.73-79.

Representations of Australia and its Soldiers essays

Representations of Australia and its Soldiers essays Different people think differently of Australia and its soldiers in the First World War. They have been depicted differently by many sources. It is obvious that there is a general opinion that the Australian soldiers at Gallipoli were eager, determined and very brave. But some have described them in a very different way. In the film Gallipoli, the character of Archy is seen to be a determined and brave young man who lost his life in what seemed a useless attack on the Turks. Frank is a cautious and sometimes bad tempered man who is also very brave and had to run through places where snipers could easily get him. He doesnt die but did a lot to help the Australians. These two, though, also had some bad points. They scratched their names on the pyramids, Frank started to argue with a shopkeeper and Archy shouldnt have been at the war at all because he was underage. In a book called Australia and the Great War, one extract says that Australians were riotous, undisciplined, loutish and disobedient. They supposedly had wild donkey ides, riots in Brothels, threw drivers off trams, scratched their names on the pyramids and had fights with shop owners. In a newspaper article, Australian soldiers are said to have made an inspiring scene in which to make her European debut as a fighting unit of the Empire. They say that after already doing this they should get all the Australians back home. In Gallipoli, once again, you get the idea that it was the Britishs fault for the loss of so many lives in Gallipoli. The British were said to be drinking tea when the Australians were fighting. The officer who order the continuation of the assault at The Nek has a British accent in the movie when he was actually Australian. This shows that many people think that in was the British who killed the Australians. Most sources tell of Australia as a country with a red, barren and harsh environment. Most people then onl...

A Guide to Vertebrates and Invertebrates

A Guide to Vertebrates and Invertebrates Animal classification is a matter of sorting out similarities and differences, of placing animals in groups and then breaking those groups apart into subgroups. The whole endeavor creates a structure- a hierarchy in which the large high-level groups sort out bold and obvious differences, while the low-level groups tease apart subtle, almost imperceptible, variations. This sorting process enables scientists to describe evolutionary relationships, identify shared traits, and highlight unique characteristics down through the various levels of animal groups and subgroups. Among the most basic criteria by which animals are sorted is whether or not they possess a backbone. This single trait places an animal into one of just two groups: the vertebrates or the invertebrates and represents a fundamental division among all animals alive today as well as those that have long ago disappeared. If we are to know anything about an animal, we should first aim to determine whether it is an invertebrate or a vertebrate. Well then be on our way to understanding its place within the animal world. What are Vertebrates? Vertebrates (Subphylum Vertebrata) are animals that possess an internal skeleton (endoskeleton) that includes a backbone made up of a column of vertebrae (Keeton, 1986:1150). The Subphylum Vertebrata is a group within the Phylum Chordata (commonly called the chordates) and as such inherits the characteristics of all chordates: bilateral symmetrybody segmentationendoskeleton (bony or cartilaginous)pharyngeal pouches (present during some stage of development)complete digestive systemventral heartclosed blood systemtail (at some stage of development) In addition to the traits listed above, vertebrates possess one additional trait that makes them unique among chordates: the presence of a backbone. There are a few groups of chordates that do not possess a backbone (these organisms are not vertebrates and are instead referred to as invertebrate chordates). The animal classes that are vertebrates include: Jawless fish (Class Agnatha)Armored fish (Class Placodermi) - extinctCartilaginous fish (Class Chondrichthyes)Bony fish (Class Osteichthyes)Amphibians (Class Amphibia)Reptiles (Class Reptilia)Birds (Class Aves)Mammals (Class Mammalia) What are Invertebrates? Invertebrates are a broad collection of animal groups (they do not belong to a single subphylum like the vertebrates) all of which lack a backbone. Some (not all) of the animal groups that are invertebrates include: Sponges (Phylum Porifera)Jellyfish, hydras, sea anemones, corals (Phylum Cnidaria)Comb jellies (Phylum Ctenophora)Flatworms (Phylum Platyhelminthes)Mollusks (Phylum Mollusca)Arthropods (Phylum Arthropoda)Segmented worms (Phylum Annelida)Echinoderms (Phylum Echinodermata) In total, there are at least 30 groups of invertebrates that scientists have identified to date. A vast proportion, 97 percent, of animal species alive today are invertebrates. The earliest of all animals to have evolved were invertebrates and the various forms that have developed during their long evolutionary past is highly diverse. All invertebrates are ectotherms, that is they do not produce their own body heat but instead acquire it from their environment.

Diffusion and Osmosis Lab Report Example | Topics and Well Written Essays - 2500 words

Diffusion and Osmosis - Lab Report Example The cell and its extracellular environments are fluid in that concentrations of substances dissolved in it vary. Depending on varying concentration of the substances dissolved in water, be it extracellular or intracellular, dissolved substances or water move from one compartment to another directed by physical principles of gradients. To accomplish this, the cell utilizes the processes of osmosis and diffusion as appropriate. These phenomena can be visualized outside the cell by simulating these in vitro. Since cell membrane is a semipermeable membrane and exchanges occur through this, these experiments must utilize a construct of the semipermeable membrane that can be made with a commercially available dialysis tubing and bag. These membranes have microscopic pores through which small molecules like water can pass, but larger molecules, such as sugar cannot pass through them. Thus through this molecular net, solutions of different solute concentrations can be observed to pass in a m easurable fashion. This experiment has been designed to prove the hypothesis of the mechanisms of osmosis and diffusion through such a semipermeable membrane. Diffusion is defined as the movement of molecules from a site of higher concentration to that of a lower concentration. Technically speaking, this difference in concentration creates a concentration gradient, and the steepness of the gradient determines the rate of diffusion. In an attempt to find out the reason as to why it occurs, it can be attributed to a great deal of space between the molecules of all substances that are candidates for diffusion. Liquids have moderate intermolecular space. Moreover, all molecules are in a state of constant random movement so that they collide and intermingle. It is expected that in solids diffusion would occur slowly for tight packing of the molecules. Liquids and gases would diffuse freely because their molecules are spaced widely. Thus, it can be stated that any solute will tend to uniformly occupy the entire space available to it. This movement, known as diffusion, is due to the spontaneous Brownian random movement that all molecules experie nce. The net result of diffusion is the movement of substances according to their difference in concentrations, from regions of high concentration to regions of low concentration. Diffusion is an effective way for substances to move short distances. Diffusion across a membrane has no preferential direction; it can occur from the outside of the cell toward the inside or from the inside of the cell toward the outside. It is however determined the extend of permeability through a membrane, which in the case of a living cell is the protein-lipid-protein plasma membrane, and therefore, diffusion across the membrane usually implies that the diffusing solute enters the lipid bilayer to cross it, the solute's solubility in a lipid solvent compared with its solubility in water is important in determining its permeability through it. Hydrophilic substances, such as ions and sugars, do not interact well with the lipid component of the membrane, hence would diffuse across the membrane more slow ly. This is, however, not applicable in physical membrane such as a dialysis membrane (Campbell NA, Reece JB, Mitchell LG., 1999). If equal volumes of pure water and a strong sugar solution are taken, the pure water will have more water molecules and would have higher water concentration that a sugar

Client and Policy Failure Assignment Example | Topics and Well Written Essays - 250 words

Client and Policy Failure - Assignment Example By doing so; I will able to cater for the interests of my clients who are affected negatively by the policy on basements and city building codes. My responsibility is to assist clients obtain affordable housing that are safe and habitable, as opposed to expensive houses that lead to people living in the streets. Thus, the clients’ get affordable housing and the city remain free of homeless people. The action-forcing event that triggered the study was the numerous complaints made by New York residents, demanding that action to be taken concerning illegal and unsafe apartments. These individuals are taxpayers being forced out of their homes, and owners of basement apartments being asked to dismantle their rental units, as in the case of Rakha and Mahbub who were asked to pay a penalty of $1,200, after spending more than $4,000 upgrading a cellar apartment of their home in Queens. In addition, the supporters of legalization of basement apartments such as Jerilyn Perine, a former city housing commissioner, and current Executive Director of the research group Citizens Housing and Planning Council claim that the issue of basement housing and other illegal houses has to be dealt with, as well as the allowance of a better city planning strategies. This is in relation to the words uttered by the City Councilman Brad Lander who said he was outlining legislation in favor of the â€Å"a ccessory† housing units, and building code standards for them. The actual policy targeted for reform is the New York City Building or the Housing Maintenance Code, (ARTICLE 5- Occupancy of Cellars and Basements). That states that basements, cellars, and â€Å"granny flats,† should not be rented or occupied due to health and safety measures. What policy options exist for Assistant Secretary of State for Consular Affairs Janice L. Jacobs to decrease employment visa fraud committed by U.S. based high-tech companies in response to the recent discovery of visa fraud committed by

Genetic transformation and competence in e. coli when exposed to the green fluorescent protein

Genetic transformation and competence in e. coli when exposed to the green fluorescent protein INTRODUCTION: The goal of this experiment was to successfully insert the plasmid pGLO, which carries genes for resistance to ampicilin and for green fluorescent protein (GFP), into competent E. coli cells thereby genetically transforming E. coli to have those specific traits. Green Fluorescent Protein comes from the jellyfish Aequorea Victoria and it emits green light when excited by blue light and when in the presence of the sugar arabinose. This protein has proven significant as a gene marker as well as other forth coming uses in biochemistry, cell and microbiology (Allison, Sattenstall, 2007). In a study done by Allison and Sattenstall (2007), it was found that introducing GFP into a cell causes changes in the cell physiology that might lead to antimicrobial susceptibility of the cell. This could be of concern because of its widespread use and Allison and Sattenstall urge caution when interpreting data from studies that used GFP (Allison, Sattenstall, 2007). According to Tsen et al., the E. c oli bacteria can naturally transform with inserted plasmids and integrate them without special treatments. As long as the DNA in the plasmids is Concatemeric linear, monomeric circular or supercoiled forms of plasmid, they can transform the E. coli, whereas linear monomer cannot transform it (Tsen, et al., 2002). The uses of green fluorescent protein in competent cells such as E. coli as mentioned before are very useful in gene markers and other studies in biochemistry, cell and microbiology, however, there are still being advances made for GFP. In a study done by Torrado, Iglesias and Mikhailov, techniques were improved in how well cells expressed the GFP gene based on the growing environments (Torrado, Iglesias, Mikhailov, 2008). In our experiment, we postulate that the E. coli will be competent for the pGLO plasmid carrying GFP and resistance to ampicilin. METHODS AND MATERIALS: In this experiment we will be using the plasmid pGLO which has genes for GFP and resistance to ampicilin. In order to force the plasmids into the E. coli cells, we will be using a heat shock treatment. This heat shock treatment causes the pores in the E. coli cell membrane to open, which allows the plasmid pGLO to enter the cell. We will test an E. coli centrifuge tube that has been exposed to pGLO and a centrifuge tube that has not been exposed to pGLO as our control. We labeled two centrifuge tubes with +pGLO and -pGLO to represent which tube carried the plasmid and which was our control, respectively. We first pipette 250 microliters of transformation solution into the tubes and added approximately 2 pen tip sized E. coli colonies. We then added the pGLO plasmids to the tube labeled +pGLO and rested both tubes on ice. The ice will make it so that the heat shock will have a greater affect on the E. coli, thereby increasing our chances of successful entrance of the plasmid. We then applied the heat shock treatment to the two centrifuge tubes by putting them in a 42ÂÂ °C water bath for 50 seconds. Afterward we put them back into the ice bath and prepared to put them into the four prepared agar plates. The four agar plates were split into two sets, two help +pGLO and two held our control -pGLO. The first plate contained just Luria Broth (LB) and 250 microliters of the -pGLO substance. The second control plate was LB with a mix of ampicilin (amp) and -pGLO E. coli solution which will ideally not grow any E. coli because E. coli by itself is not resistant to ampicilin. The third plate is used with +pGLO solution and is another LB/amp plate. The fourth and final plate is another +pGLO plate and contains LB and ampicilin but also arabinose, which is needed for the expression of the GFP. RESULTS: In this experiment, we used heat shock treatment in order to insert the pGLO plasmid into E. coli cells because the plasmid carries the genes that code for green fluorescent protein and ampicilin resistance. After a week of incubation in the refrigerator, we analyzed our four Petri dishes. All plates came out as predicted. Our first control plate (-pGLO E. coli cells) contained Luria Broth and ampicilin and it sustained 0% E. coli cell growth. The second control plate (-pGLO E. coli cells) contained only Luria Broth and there was 100% coverage of the agar plates. The lawn made by the E. coli cells was a whitish clear color in normal light and were not fluorescent green when exposed to UV light. In the two transformation plates, we received positive results matching our predictions. In the first transformation plate (+pGLO E. coli cells) there was a presence of Luria Broth and ampicilin. There were roughly 140 colonies of E. coli cells which was about 60% coverage of the Petri dish. Each of the colonies was an off whitish color under normal light but were not fluorescent green under UV light. The second transformation plate (+pGLO E. coli cells) contained a mixture of Luria Broth, ampicilin and arabinose. This plate had E. coli colony growth but there were only roughly 40 E. coli colonies, making about a 25% coverage of the plate. Again these colonies were whitish in color when exposed to normal light however, they did turn fluorescent green under the UV light. TABLE OF RESULTS: TYPE OF PLATE CONTENTS SKETCH OF PLATE OBSERVATIONS Transformation +pGLO/LB/amp/ara -Growth of Colonies (40 count; 25% coverage) -Whitish color in normal light -Fluorescent green under UV Transformation +pGLO/LB/amp -Growth in Colonies (140 count; 60% coverage) -Whitish color in normal light -No fluorescent green color under UV Control -pGLO/LB/amp -No E. coli growth, E. coli not transformed (0% coverage) Control -pGLO/LB -E. coli growth present (100% coverage) -Whitish color in normal light -No fluorescent color in UV light DISCUSSION: The hypothesis is the following: After heat shock treatment, the competent E. coli cells will receive the plasmid pGLO, and the E. coli cells will be transformed. We predicted that the E. coli cells would take in the plasmid and transform in our two transformation plates. In the -pGLO/LB control plate we predicted that significant growth would happen because there is no antibiotics and only an optimal growing environment. In the -pGLO/LB/amp control plate we predicted that there would be no growth of E. coli because ampicilin is present, an antibiotic that E. coli is not naturally resistant to. In the transformation plate +pGLO/LB/amp we predicted that there would be E. coli growth considering we hypothesized that the plasmid would be accepted by the E. coli cell, thereby giving it ampicilin resistance. In our last transformation plate +pGLO/LB/amp/ara we again expected growth of E. coli since we hypothesized the E. coli cell would be competent for the plasmid. We also expected that this would be the plate to glow fluorescent green since arabinose, the sugar that allows for the glowing, was present in the agar plate. In order for this experiment to show true results, we added the two control plates with different purposes. The first plate contained only Luria Broth, the ideal growing environment for E. coli. This plate was used to make sure that our E. coli cells were healthy and able to grow consistently. If they were unable to grow, that would mean that our cells were unhealthy or contaminated, which would in turn affect the results of our transformation plates. Our results for this plate were that we had healthy E. coli cells since they produced a full lawn. Our second plate was the one with both Luria Broth and ampicilin for the growing environment. We did not have any growth of E. coli on this plate, just as we predicted. This is good because the plasmid we were using to transform the E. coli cells have the gene that causes ampicilin resistance. If our E. coli had been contaminated or already transformed from its non-resistance state, we would see it in this control plate. If we had seen gro wth, we would know that our results for the transformation plates were faulty because our normal E. coli was already resistant. Next we examined our transformation plates. These plates were the ones that we exposed to the pGLO plasmid. Our first plate had Luria Broth and ampicilin, just like our control; however, since we treated this batch with the pGLO plasmid followed by heat shock treatment, we expected to see growth. Our results from this plate did show that the E. coli grew in the ampicilin agar plate, thereby showing how many of the E. coli cells received the plasmid and were able to be genetically transformed. However, under the UV light, the colonies did not glow fluorescent green because of the absence of arabinose. Our second transformation plate had Luria Broth, ampicilin and arabinose. Our results followed our predictions that we would see growth and have the colonies glow under the UV light. This is because the E. coli that took the plasmids were transformed so they showed their new resistance to ampicilin and they showed that when grown in an environment where arabinose is present, the green fl uorescent protein will be expressed. Our results did indeed support our hypothesis because the E. coli were transformed in our transformation plates because we saw that colonies were able to grow in an environment where ampicilin was present and also the green fluorescent protein was expressed in arabinose rich environments. There was an area of weakness in our experiment. The crucial part, the heat shock that opens the cellular membrane pores, could have gone a bit smoother. Our times were not exact due to many groups trying to do this part all at once. Secondly, there is room for error in the consistency of our experiment plates since each member of the group took turns at each stage of the process. In conclusion, the results of our experiment proved our hypothesis that the E. coli cells were competent for the pGLO plasmid. Our results were consistent with our predictions. We found that the E. coli cells can be transformed by the plasmid after our heat shock treatment. Our control plates can out controlled and our transformation plates produced colonies that expressed the GFP gene. SOURCES: Allison, D.G., Sattenstall, M.A. (2007). The Influence of green fluorescent protein incorporation on bacterial physiology: a note of caution. Journal of Applied Microbiology, 103(2), 318-324 Suh-Der Tsen, S., Suh-Sen Fang, S., Mei-Jye Chen, S., Jun-Yi Chien, S., Chih-Chun Lee, S., Han-Lin Tsen, D. (2002). Natural Plasmid Transformation in Escherichia coli. Journal of Biomedical Science, 9(3), 246-252. doi:10.1159/000059425. Torrado, M., Iglesias, R., Mikhailov, A.T. (2008). Detection of protein interactions based on gfp fragment complementation by fluorescence microscopy and spectrofluorometry. BioTechniques, 44(1), 70-74.

Jazz Performance Essay

Two weeks ago, I attended a jazz performance in a jazz bar together with a few close friends. It was a two hour show and there were 3 sets. The band was made up of five members. One played the drums, another was on piano, two played wind instruments, and the other served as the lead singer. Their performance was very alive and truly entertaining. They accepted requests from the guests. The band had a wide repertoire. Mostly, the songs they played were derived from the blues era. However, one set was devoted to recent jazz hits such as those of Michael Buble. Some of the songs they played were those of Marvin Gaye, Louis Armstrong, and Kenny G. Also, they devoted majority of their third set to Bossa Nova sound. The band did not play any of their own compositions. Being that they were only formed six months ago, they have yet to reach the point wherein they can come up with their own songs. Their show was truly entertaining. They brought a lot of brightness to an otherwise dull venue. There was not much lighting in the bar. The spotlight was directed towards the stage. I believe such lighting was meant to emphasize the intimacy of the event and of the place. Majority of the crowd was composed of couples. The music they played was ideal for dates. The music they played enhanced the romantic mood inside the venue. There was no political symbolism in their music. The main purpose of their music was to entertain the guests. Admittedly, there was nothing truly special about their performance. In fact, it can be considered a regular gig for a regular band. However, the important element in their performance was that they were able to entertain the guests and that they were able to make the people enjoy their time listening to their music.

Modification Of The Juvenile Justice System And The Affect...

Modification in the Juvenile Justice System and the Affect on the Future of our Youth Michelle Fleeton Auburn University at Montgomery Abstract The Juvenile Justice System (JJS) has transformed tremendously over the last 300 years. From adolescents and adults being charged with the same brutality, to the introduction of rehabilitation and diversion programs. As time went on and new political policies were brought forward in the courts, the crime rates fluctuated until the late 1900s and with policies like the Gun Free School Act (GFSA) the number o incarcerated delinquents skyrocketed. Though the conditions of juveniles that are incarcerated have somewhat improved with Positive Youth Development (PYD), there is still a large amount of discrimination in the JJS today. The system is failing to ensure that all youth with disabilities, mental issues, lower class and minorities are provided with the resources needed to live a productive and crime-free life outside of jail. Juveniles are the future of our country and it is imperative that they receive the proper education needed to become effective adults in our society. The JJS n eeds to be restructured to more effectively lower the chances of recidivism and advocate higher education programs for our youth. In order to accomplish this, there has to be complete reformation of PYD and restorative justice. Keywords: delinquent, court, restorative, punishment, juvenile justice, law, criminal, development, rehabilitation.Show MoreRelatedRevealing the Injustices of America and Its Continued Perpetuation in The Narrative of the Life of Frederick Douglass1530 Words   |  6 Pagesand there is no hope for any betterment, unless he escapes to freedom. While this situation seems horrific to modern-day eyes, it is worth asking if our present circumstances are really so different, given the class disparities that exist within America today. 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The Box, Inc. - 1530 Words

Introduction Jack in the Box, Inc. opened its first restaurant in 1951. Today, it has become one of the nation’s largest hamburger chain and â€Å"based on number of restaurants, is the second largest QSR hamburger chain.† In addition to its QSR hamburger chain, Jack in the Box â€Å"acquired Qdoba Restaurant Corporation, operator and franchisor of Qdoba Mexican Grill.† 1 According to its 10-K report, Qdoba â€Å"is the second largest fast-casual Mexican brand in the United States.† 1 Jack in the Box, Inc. offers a variety of products on their menu. Products that vary from hamburgers, tacos, specialty sandwiches, drinks, real ice cream shakes, salads and side items. The customer’s ability to customize their order to their specific taste preferences is one of many core competencies. Along with the option of customization, its other core competencies is their drive-thru. 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