Therapeutic Proteins †Hirudin

Therapeutic Proteins – Hirudin Isaac Johnson Hirudin is a polypeptide that has anticoagulant properties (Rosenthal, 2008). Hirudin is important because it is a direct thrombin inhibitor. Since the presence of thrombin, a serine protease, is indicative of a blood clot having a polypeptide that specifically inhibits thrombin generation is important in a number of ways (Contributors, Thrombin, 2013). Hirudin is used to prevent the formation of blood clots in arteries, veins, and chambers within the heart (Rosenthal, 2008). Hirudins produced by recombinant biotechnology are more effective than other serine proteases produced by the body because they break up several different types of thrombin and they don’t bind to plasma proteins, which means there is a known dose-response relationship (Andreas Greinacher, 2008). The clinical use of r-hirudin provided an alternative choice to those people who experienced complications when given heparin; in addition â€Å"r-hirudin is important because it binds directly to thrombin, there by forming a complex with thrombin not only at its fibrinogen-binding site but also at its catalytic region†(Eichinger, 1995, p. 886), which completely stops the generation of thrombin unlike heparin which stops the production of thrombin by â€Å"catalyzing the inactivation of thrombin using anti-thrombin III†(Eichinerger, 1995, p. 886). Hirudin was discovered by John Haycraft in 1884 during his research on the coagulation of blood. In the 1900s it was extracted from leeches but today it is produced through recombinant biotechnology (Contributors, Hirudin, 2014). â€Å"The first recombinant hirudin produced and approved for clinical use, called lepirudin, was approved by the FDA in 1998 for the treatment of heparin-induced thrombocytopenia complicated by thrombosis† (Greinacher, 2008, p. 819). Heparin-induced thrombocytopenia is a disorder which causes clotting to occur instead of excessive bleeding and is countered by administering another anticoagulant to inhibit thrombocytosis (Contributors, Heparin-induced thrombocytopenia, 2013). Hirudin is a polypeptide produced by the parapharyngeal glands of the leech, Hirudo medicinalis. Hirudins produced through recombinant biotechnology are generally produced using Pichia pastorus although Escherichia coli can be used as well albeit with the risk of degradation (Wuguang Lu, 2012). The process of producing recombinant hirudin (rHIR) is a multi-stage process, the main stages are: Construction of HIR/pPic9k, Transformation of P. pastorus strain, Shake flask evaluation of transformants, Clone fermentation, rHIR purification (Stuart A. Rosenfeld, 1996). The first stage, the construction of HIR/pPic9k, had several steps: use of pLMFÃŽ ±1 as a template for the PCR amplication of the hirudin gene, the use of primers to clone the gene, and the use of restrictive endonucleases to cut out part of the vector so that the hirudin gene could be inserted into pPic9K vector. Because the hirudin gene was inserted into the expression vector it allowed for greater control of the cloning process and allowed for optimization of the process. In the second stage, the transformation of P. pastorus strain, the expression vector was digested with Sa1I, a restrictive endonuclease, and the DNA was integrated into the host genome through homologous recombination via electroporation. Because the Hir/PPic9K was digested with Sa1I a large amount of His+ transformants were obtained after electroporation. These transformants were replica-plated, in order to obtain resistance to the antibiotic G418, whereupon the transformants could then undergo multiple chromoso mal integrations of the HIR gene. In the third stage, the shake flask evaluation of transformants, the transformants were screened to determine if they were capable of expressing and secreting recombinant hirudin. All of the transformants from the experiment did secrete recombinant hirudin albeit at different rates over the course of the 7 day evaluation. In the fourth stage, clone fermentation, the cultures were fermented in order to increase production of recombinant hirudin. Fermentation conditions increased r-hir production based on dissolved oxygen content, nutrient balance and high cell densities which made the environment optimal for recombinant hirudin production. The fermentation process was a batch process and used glycerol as the sole carbon source followed later by feeding the cultures methanol to fully induce rHIR expression. The methanol feed rate was varied so that a low concentration could be maintained in the fermentor thus keeping toxicity levels low. A low pH leve l is required for the fermentation process; a pH of 5 is too high and causes the proteins to break down while a pH of 3 is optimal for rHIR production. The final stage, the purification of rHIR, used a two-step purification protocol where first Q-Sepharose ion exchange chromatography was used followed by preparative high-performance liquid chromatography (HPLC). The recombinant hirudin had a purity percentage of more than 97% and a recovery yield of 63% (Stuart A. Rosenfeld, 1996). The impact of therapeutic proteins on society has been overwhelmingly positive. Recombinant proteins have many uses in society and have been incorporated into many different facets of everyday life. They are used in many different types of therapy from growth hormone treatment in adults to the use of recombinant interferon used to treat Hepatitis C. Recombinant proteins don’t just impact the medical field though they also play an important role in cosmetics and as dietary supplements. Furthermore when recombinant proteins are compared to blood plasma-derived products they are clearly shown to be safer. Since recombinant proteins were first introduced in the early 1990s there have been few adverse effects caused by them and zero fatalities whereas blood plasma-derived products carry the risk of transmission of emerging diseases that could turn into epidemics like the swine flu or bird flu (Liras, 2008). In conclusion hirudin is a polypeptide that is a direct inhibitor of thrombin and thus is an excellent anticoagulant. Several different types of recombinant proteins derived from hirudin have been approved for clinical use since the late 1990’s. These alternatives, like lepirudin, fill a necessary niche in the medical world that other coagulants cannot fill due to either patient complications with those drugs or hirudin derived drugs simply being more effective. The role that therapeutic proteins play in medical treatments and therapy will increase in society at they become more cost effective and production methods become more efficient. References Andreas Greinacher, T. E. (2008). The direct thrombin inhibitor hirudin . Thromb Haemost, 819-829. Contributors, W. (2013, November 2). Heparin-induced thrombocytopenia. Retrieved January 11, 2014, from Wikipedia, The Free Encyclopedia: http://en.wikipedia.org/wiki/Heparin-induced_thrombocytopenia Contributors, W. (2013, December 23). Thrombin. Retrieved January 11, 2014, from Wikipedia, The Free Encyclopedia: http://en.wikipedia.org/wiki/Thrombin Contributors, W. (2014, January 4). Hirudin. Retrieved January 11, 2014, from Wikipedia, The Free Encyclopedia: http://en.wikipedia.org/wiki/Hirudin Liras, A. (2008). Recombinant proteins in therapeutics: haemophilia treatment as an example. International Archives of Medicine, 1-4. Rosenthal, S. O. (2008). Encylopedia of Molecular Pharmacology. New York: Springer-Verlag Berlin Heidelberg. Sabine Eichinger, M. W.-L.-G. (1995). Effects of Recombinant Hirudin (r-Hirudin, HBW 023) on Coagulation and Platelet Activation In Vivo. Arteriosclerosis, Thrombosis, and Vascular Biology, 886-892. Stuart A. Rosenfeld, D. N. (1996). Production and Purification of Recombinant Hirudin. PROTEIN EXPRESSION AND PURIFICATION Article No. 0127, 476-482. Wuguang Lu, X. C. (2012). Production and Characterization of Hirudin Variant-1 by SUMO. Springer Science+Business Media, 41-48.

biology presentation for the skeleton :: essays research papers

Intro – slide 1 – acetate 1 The skeleton of a human foetus is formed from tough but flexible cartilage that acts as a blueprint for bone construction. During ossification ( the changing of cartilage to bone) which begins before birth, the cartilage is broken down and the resulting space is filled by bone building mineral salts and protein fibres secreted by bone cells. Humans have a bony endoskeleton made up of 206 bones, although we are born with up to 300, but many of these fuse during childhood. slide 2 The skeleton is divided into two parts ; the axial and the appendiculur. Axial skeleton The axial skeleton consists of the skull, backbone and rib cage which forms the upright axis of the body. It helps to protect the brain, spinal cord and organs in the chest. Appendicular skeleton The appendicular skeleton consists of the upper and lower limbs and the pectoral (shoulder) and pelvis and girdles. The human pelvis is adapted for an upright stance. the lower limbs support the upper body and enable walking and other locomotory movements to take place, where as the upper limbs are used for manipulation. Slide three Just read off screen Slide four – acetate 2 Muscles are used to move your bones. Most muscles are joined at both ends to bones. They are joined to bones by non elastic tendons. Bones are held in place by strong fibres called ligaments. Slide five – acetate 3 Joints are formed when two or more bones come together. Most joints allow bones to move. The amount of movement depends on the type of joint. The elbow joint is a hinge joint. Joints that allow movement are called synovial joints. A synovial joint s adapted to enable a joint to move easily. They contain cartilage, ligaments, bones, synovial fluid and a synovial membrane. Slide six The bones provide the solid base on which the tendons and ligaments are fixed. They are very resistant to being compressed, bent and stretched. The cartilage is the smooth layer which covers the ends of the bones and which stops the bones rubbing together. It has high tensile strength, but it is not rigid. Can compressed and is able to act as a shock absorber. The ligaments are strong fibres that hold bones firmly together. they form a protective cover around the joint. Are very strong and sufficiently elastic to allow movement when the bones in the joint move, so reducing the chance of dislocating a joint.

Aol Time Warner- What Went Wrong

In January 2000, AOL announced that it would be acquiring Time Warner through a complete stock deal to create the largest media company in the world. Not only was the merger the biggest ever in the media industry, it was also one of the biggest in the history of the corporate world. As per the merger agreement, AOL and Time Warner stock was converted to AOL Time Warner stock. AOL shareholders received one share of AOL Time Warner for each AOL share owned and Time Warner shareholders received 1. shares of AOL Time Warner for each Time Warner share they owned. While AOL shareholders owned 55% of the new company, the remaining was held by Time Warner. The merger was soon being talked of as the beginning of a new trend: the coming together of traditional and new media companies. According to a report, AOL was ‘a turbo-charged engine' that would bring old media giant, Time Warner, into the Internet century. The merger was expected to result in a 30% increase in profits, amounting to over US $ 40 billion in revenues in the first year itself. The new company had 100 million paid subscribers, which included the customers of AOL's dial-up service and subscribers of Time Warner's cable and magazine divisions. A major setback to the success of the merger was the bursting of the Internet bubble, which was expected to rule the media and entertainment industry in the 21st century. When the Internet bubble burst, there was a steep decline in subscriber growth for AOL, which led to a steep decline in its advertising revenues. Time Warner merged with America Online in 2001 at the height of the dot-com boom, with AOL using its inflated stock as a currency for the transaction. But the marriage of old and new media behemoths baptised quickly went sour as the benefits promised to shareholders failed to materialise. AOL was valued at more than $US150 billion when the ill-fated merger was announced, but its worth collapsed dramatically as the dot-com bubble burst. Time Warner was forced in 2002 to massively write down the value of the Internet unit and the AOL name was removed from the group's corporate title in 2003. As a consequence of this not only lost AOL subscribers to their Internet service but also their portal lost importance leading to a loss in opportunity to promote AOL Time Warner content. As a further consequence income from advertising is decreasing. 3. 0 BCG Matrix of AOL-TIME WARNER 4. 0 ALTERNATIVES REVIEW AND TURNAROUND STRATEGIES After the merger, AOL Time Warner was indeed the unquestioned number one worldwide media company, especially with the phenomenal box-office successes of Harry Potter and Lord of the Rings. Some of the anticipated synergies have actually come about and the company has successfully met a number of its goals, according to industry observers. On the Internet side, AOL must prove it can maintain its paramount position by signing deals across the entire cable industry while it opens its Time Warner cable pipes to competitors. If it can’t, AOL’s position as the undisputed leader in the consumer Internet business could begin to slip. According to published reports, AOL has more than 33 million subscribers worldwide, 25 million in the U. S. and analysts expect AOL to sign 6. 2 million new members for the year. AOL’s Pittman believes that eventually between 90 million and 95 million U. S. homes will be connected to the Internet. In 2001, AOL Internet subscribers spent $33 billion online, 67% more than they spent the year before. America Online has thrived by making it cheap and easy for millions of people to get onto the Internet using a standard open-access phone line and modem. With all of the hype about broadband, nearly all of AOL’s current subscribers still use dial-up â€Å"narrowband† connections, which are far too slow to accommodate the growing demand for rich media that consumers increasingly want from the web. 4. 1 AOL-TIME WARNER TURNAROUND STRATEGY The problem faced by Time Warner after its merger with AOL is an issue which merits discussion. The AOL-Time Warner merger in 2001 resulted in the largest media company in the world. AOL joined hands with Time Warner to create synergy between its online businesses and Warner's media business. Two significant factors affected the post merger company. One, the dot-com burst meant adverse effect on AOL’s advertising revenues. And two, dial-up subscribers decreased thereby affecting revenues and overall profitability of AOL. A key element in the turnaround strategy should be to offer free content on its portal. This strategy will benefit AOL in attracting more online users and advertising revenues. When AOL began operations it soon became the leading company for-pay online subscriber service, bringing easy-to-use Internet service to more than 30 million users. AOL was mainly based on around its dial up business. With customers shifting to broadband, AOL was losing subscribers rapidly. The dial-up segment though profitable, was declining in revenues having lost 2. 6 million subscribers in a period of one year. The share price of AOL Time Warner fell by 60% after the merger. The merger was heavily criticized from all quarters. Growth in advertising business came with AOL establishing itself as a support service rather than an internet access provider. Seeing AOL’s success Google entered into a global advertising partnership with the AOL. Google acquired a 5% equity stake in AOL for US$ 1 billion. In broadband, to be able to beat the competition they should be the first organization to define the category of streaming broadband, bringing relevant content and relevant programming into the broadband household so as to let people search for the video and music they want to view and listen to. Provision of other a la carte and premium services, such as being a place for people to store pictures, documents and e-mails they create or receive while they're on the Web. It makes sense. What people want is all their assets available all the time, regardless of where they are. This is especially important for multichannel users, who will be a larger and larger portion of our customer base. Part of the day, people will need to access AOL from their laptops or at home on high-speed DSL or cable, or a smart phone or communication-enabled PDA. The organization should be re-energized. There should be a reaffirmed commitment, a new management team on the field. Positioning the brand and regaining trust are all smart things to do and it will serve as the litmus tests for the decisions to be made. Now, they must be able to recognize emerging trends and stake out our role in them before they take hold among consumers. The overall success of this business long-term will be their ability to integrate best-in-class technology and programming and create compelling value for each of its targeted customer segments. As a technologist, could you ask for a better role? Every decision matters. 5. 0 CONCLUSION The credo of the combined AOL Time Warner was synergy, the idea that the companies would be greater than the sum of their parts because each of the divisions would help market the others. But the dream never materialized. The fast-growing AOL online service was supposed to be the juggernaut that would cause the whole company to grow at 30 percent a year. It turned out that AOL was unable to sustain its own growth rate, much less carry Time Warner. In the end, however, the true success of the AOL Time Warner merger will be won or lost, on the company’s ability to leverage its promising collection of cable networks into something more than just a way to see the latest episode of miniseries. That will only be done by bringing AOL’s online expertise into alignment with Time Warner’s stable of media properties, and adding a broad vision of the next chapter of the Internet. This means being the first to create an invisible high-speed grid to deliver interactive services and entertainment to consumers without muss or fuss †¦ anywhere, anytime, over any device. If AOL Time Warner can do that, the company may be able to create the most ironclad and lucrative relationship with the consumers of the future. â€Å"

Personality Theory Bandura Carl Rogers - 3322 Words

Running head: Albert Bandura Carl Rogers Compare and Contrast Social Cognitive and Humanistic Theories of Personality to the case study entitled Myesha Course: PSCY3017 Personality Theory II Personality is an intriguing component in psychology vital for the perception of human beings. Understanding and defining personality has proven to be a difficult task. It is so complex, in fact, that no single theory can adequately define it. If one was to ask an ordinary individual to do so, some of the most common answers might be â€Å"a person’s characteristics† or â€Å"the impression (s) one makes on others†. Personality Theorists on the other hand view personality as the essence of the person, the individual’s†¦show more content†¦For instance Myesha quickly realized that Law was not for her. She considered dropping out of the University as she had gotten involved with a spoken word group around this time. As a result, her stepfather threatened to stop supporting her if she dropped out and this placed a strain on family relationships. These are all factors of cause and effect, which are influencing each other. Bandura views Myesha as an agentic operator in her life. He would believe that she has the capability to intentionally make things happen by her actions and that she is not an â€Å"onlooking host of internal mechanisms orchestrated by environmental events† (Bandura, 2001, p.2). Individuals are sentient agents of experiences rather than simply undergoers of experience. The sensory motor and cerebral systems are tools people use to accomplish the tasks and goals that give meaning and direction in their lives (Harrà © Gillet 1994). Carl Rogers also agrees to some degree with Bandura, in the belief that the environment also affects us and the people in our environment determine what our behavior will be like (Pervin, Cervone John, 2005). He also believes that her experience in the spoken word group can have an impact on her personal growth and individual experiences. The belief is that Myesha’s experiences are unique, and that her perception of the world is critical to understanding and achie ving a particular behavior that would be identical to her becomingShow MoreRelatedIs Personality Based On Science?986 Words   |  4 PagesIs personality based on science? Is personality a science? Precisely, what is personality? There has been many theories about personality as it has been viewed differently over the years. With personality, no two theories are the same just as no two individuals are. Personality is an individual’s characteristic pattern of thinking, feeling, and acting. 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