Myoglobin In The Utilization Of Oxygen In Animals Biology Essay

Myoglobin In The Utilization Of Oxygen In Animals Biology Essay Myoglobin is an oxygen-binding globular protein which is vital in facilitating the acquisition and utilization of oxygen in animals. Myoglobin was isolated and purified from ground water buffalo extract. The Myoglobin was isolated by cation-exchange chromatography, and concentration of Myoglobin is measured by spectrophotometry. Total concentration of protein was determined by performing Bradford protein assay. Iron analysis was performed by atomic absorption spectrophotometry (AAS). Molecular weight of Myoglobin was determined by running SDS-PAGE. A total amount of 144 ug Myoglobin was purified by the cation-exchange chromatography with a relative purity of 25.9%. The concentration of the purified Myoglobin was determined to be 0.072 ug/uL. The molecular weight of Myoglobin was determined to be 18204 Da And 0.423 mg of iron were detected in the acid digested extract Introduction Myoglobin is a globular protein which contains a single polypeptide chain of about 153 amino acids and an iron-porphyrin complex, or the heme group (3). And it has a molecular weight of 16700 Da and 153 amino acids (4). The non-covalently bound heme group, which resides in the hydrophobic interior of the native globin chain, is able to unfold under acidic condition and consequently weakens the interaction between the heme group and the globin (5). Myoglobin is an oxygen-transport protein which can be found in muscle tissues of all mammles. Myoglobin is critical in mammalian cell in that it is responsible for storage and distribution of oxygen, and possibly carrying energy (4). Diving animals such as seals and whales have excessive amount of Myoglobin that help them travel undersea by storing and transport oxygen (9). It plays a significant role in the physiological function of heart and skeletal muscle (2). Elevated consumption of oxygen during exercise necessitates the production of myoglobin in red muscle and heart cells, and the transportation of oxygen by myoglobin from the sarcolemma to the mitochondria in vertebrate heart and red muscle cells. (1) Spectrophotometry studies the interaction of electromagnetic radiation with molecules, atoms and ions (10). It can shed light on the physical and chemical properties by measuring the emission or absorption of electromagnetic radiation (10). Besides, it is also used to identify biomolecules from their individual absorption spectrum. In the meantime, spectrophotomoetry can quantitatively measure the concentration of molecules in solution According to the Beer-Lambert law, the fraction of incident light absorbed by a solution at a given wavelength is indicative of the concentration of the absorbing species (10) . Tryptophan and tyrosine can absorb ultraviolet light, which accounts for the characteristic strong absorbance of light at wavelength of 280 nm by most proteins (9). Atomic absorption spectroscopy (AAS)can be used to determine the identity and concentration of chemical elements in the gaseous state by measuring the light radiation absorbed by the elements (10). Ion-exchange chromatography is a technique for separating biomolecules capable of being involved in electrostatic interactions (10). Molecules can be separated based on their sign and magnitude of net charge at a given pH and formation of electrostatic linkages between thee resin and the protein of interest (9). Ion exchange separations take place in columns packed with an iron-exchange resin (10). Resins with bound anionic groups are cation exchanges whereas those with bound cationic groups are anion exchangers (9). Selection of ion exchange resin depends on what is to be purified, the pH to be used in the column, and the strength of the functional group (10). Electrophoresis is the separation of proteins based on the motion of the charged proteins under the influence of an electric field (9). The migration of the protein depends on its shape, size, charge and chemical composition (10). An electrophoretic method, polyacrylamide gel electrophoresis (SDS-PAGE) can be used to estimate the purity and determine the molecular weight of the protein (9). Treatment of the protein by the ionic detergent sodium dodecyl sulfate (SDS) can give it a uniform net charge, and protein can then be separated based solely on its mass (10). In this experiment, Myoglobin is extracted and purified by cation-exchange chromatography from ground water buffalo using Beffer A (20 nM, pH 5.6, KH2PO4) and Buffer B (20mM, Tris buffer, pH 7.5). Absorbance of the eluent fractions is measured at 280nm and 417nm. Total amount of protein and the concentration of Myoglobin are determined using Bradford assay from BSA protein. Molecular weight of Myoglobin was determined by performing SDS-PAGE. Iron content in the acid digested extract is measured by atomic absorption spectrophotometry (AAS). A total amount of 144 ug Myoglobin was purified by the cation-exchange chromatography with a relative purity of 25.9%. The molecular weight of Myoglobin was determined to be 18204 Da. 0.757 mmol of iron and 39.5 nmol of myoglobin were detected in the acid digested extract, with a ration of 20:1. Results A total of five samples were collected from a solution of 10.02g thawed ground buffalo mixed with 20.0mL, 20mM, pH 5.6, KH2PO4 (Buffer A): 1.0 mL of crude extract (sample C), 1.0 mL of filtered extract (sample D), 2.0 mL of Buffer A sample solution (sample A), 2.0 mL Buffer B sample solution (sample B) and 5.0 mL acid digested extract solution. Chromatogram for the purification of myoglobin carried out by cation-exchange chromatography shows a large peak from fraction number 5 to fraction number 12. (Figure 1). The figure also indicates another peak at fraction number 23, with smaller peaks at fraction number 19 and fraction number 25 (Figure 1). Fraction number 23 has the highest 417 nm/ 280 nm (3.766) and the highest absorbance 90.278) at 280 nm (Figure 1). Discussion When myoglobin was separated by cation-exchange chromatograph, purity of the myoglobin in the eluent fractions collected at specific volumes was examined by spectroscopy. Absorbance of Myoglobin, specifically, was measured at 417 nm and other proteins was measured at 280 nm, due to the presence of Tyrosine and Tryptophan Most proteins absorbs at a wavelength of 280 nm (10). Phophyrin has an absorbance spectrum of 414nm to 418 nm, and intensity and wavelength of the absorption can be influenced by the peripheral substituents on the porphryin and the protonation state of the nitrogen atoms (6). Sincemyoglobin consists of a iron-containing heme prosthetic group with an iron-contained porphyrin ring (9), it can absorb at a wavelength of 417. Myoglobin has a PI value of 7, so it will have a net positive charge when pH is below its PI and a net negative charge when pH is above its PI. When myoglobin is positively charged when buffer A (pH=5.6) is used to wash the column, it binds to a column containing negatively charged beads in cation-exchange chromatography. The positively charged Myoglobin can then be eluted by washing the column with buffers having higher pH value than 7 (Buffer B, pH= 7.5). Raising the pH of the mobiles phase buffer renders the Myoglobin less protonated and thus negatively charged. As a result, the Myoglobin is not be able to form an ionic interaction with the negatively charged stationary phase and then elutes from the column (10). 144 ug Myoglobin was recovered form the column, which accounts for 10.7% of the myoglobin that was loaded onto the column. Loss of Myoglobin could be attributed to diffusion spreading of Myoglobin and other contaminant proteins within the mobile phase, as a result of the increase in time length (9). Recovery of myoglobin can be improved by using narrow columns, longer column (7). The molecular weight of Myoglobin (18204 Da) obtained from the SDS-PAGE was similar to the literature value, 16700 Da (4), which indicated that the purified protein in the Buffer B sample was Myoglobin. The clear band generated from Lane B migrated the same distance as the band from the Myoglobin standard, which further confirms that the purified protein was Myoglobin (Figure 3). The stoichiometric relationship between iron and hemoglobin is 20:1 instead of 1:1 according to the fact each molecule of Myoglobin has only one molecule of ion. The excessive amount of Fe might be present in oxygen-carrying protein such as hemoglobin, which contains four heme prosthetic groups. Electron carriers in the mitochondrial respiratory chain have Fe incorporated in their prosthetic group, and examples of them include cytochomr c, ubiquinone and cytochrome oxidase. Besides, another hemeprotein, ferritin might also be present in the protein sample. Hemeprotein functions to stores and release iron atoms in biologically available form for use in heme and nonheme proteins and biochemical reactions (8). In conclusion, a ratio of 1:20 for Myoglobin and iron was obtained from the ground water buffalo. 144 ug Myoglobin was purified from the filtered extract with a relative purity of 25.9%. According to SDS-PAGE, the molecular weight of Myoglobin was determined to be 18402 Da.

Cyber Security Essay

President Obama has declared that the â€Å"cyber threat is one of the most serious economic and national security challenges we face as a nation†¦ America’s economic prosperity in the 21st century will depend on cyber security.† So why is it that so many people still don’t know some of the basic ways that produce cyber threats? The purpose of this paper is to name some of the more basic cyber threats. Even though it is just the beginning to the hazards you can obtain, they are highly effective because of the number of people that still allow these kinds of breaches. First and foremost are viruses. According to cknow.com, â€Å"There were over 50,000 computer viruses in 2000 and that number was then and still is growing rapidly. Sophos, in a print ad in June 2005 claims â€Å"over 103,000 viruses.† And, Symantec, in April 2008 is reported to have claimed the number is over one million.† With so many different viruses out there, how do you know what to look for and how to protect your cyber world from viruses? Some of the most common viruses that effect people are Trojans and Net Bots. PCmag.com defines a Trojan as â€Å"A program that appears legitimate, but performs some illicit activity when it is run. It may be used to locate password information or make the system more vulnerable to future entry or simply destroy programs or data on the hard disk. A Trojan is similar to a virus, except that it does not replicate itself. It stays in the computer doing its damage or allowing somebody from a remote site to take control of the computer. Trojans often sneak in attached to a free game or other utility.† To add to their definition, it can also be affixed to an email attachment or the email itself. So now that you know what it is you’re probably thinking so what can I do to protect myself from obtaining this virus? The best thing to do would be to educate yourself about this threat. Know what they look like and common websites they reside. The next thing that to do is to be sure to always have an up to date WELL KNOWN antivirus such as Sophos, McAfee, or Norton. The well known part is very important because some of the smaller antivirus companies are part of the reason there are so many Trojans out there. Once you get some antivirus software you need to be sure to keep it up to date. Also, don’t open emails from senders that you don’t recognize. Several of the Trojans out there that get circulated by email come from people opening emails from unknown senders. The next one is botnets. http://searchsecurity.techtarget.com says â€Å"A botnet (also known as a zombie army) is a number of Internet computers that, although their owners are unaware of it, have been set up to forward transmissions (including spam or viruses) to other computers on the Internet. Any such computer is referred to as a zombie – in effect, a computer â€Å"robot† or â€Å"bot† that serves the wishes of some master spam or virus originator. Most computers compromised in this way are home-based. According to a report from Russian-based Ka spersky Labs, botnets — not spam, viruses, or worms — currently pose the biggest threat to the Internet.† Trojans are the major way that you can get one of these viruses. So it is important that you again get antivirus software, keep it up to date, and don’t open emails from unknown senders. However, with this virus you can get it just from going to a questionable web site. So how do you know what ones are good and what ones are questionable? Well that you can for the most part guess using these deciding factors: Are there several java applications on the web page? Is there a lot of download able content? Are there pop up advertisements that come up on the page? Then you probably should do your best to stay away from the page. In case you are unsure, some antivirus software are now including a trusted and untrusted feature so when you search a site it will let you know if the page is safe or proceed at your own risk. If your antivirus does not have such a feature then there are some applications out there that will do the same thing just to name one would be WOT or Web of Trust. The next issue is a little more targeted to the business world. It deals more with their emails and correspondence but can also be directed to an average internet user. It is phishing. Webroot.com expounds â€Å"Email Phishing scams are carried out online by tech-savvy con artists and identity theft criminals. They use spam, fake websites constructed to look identical to a real sites, email and instant messages to trick you into divulging sensitive information, like bank account passwords and credit card numbers. Once you take the phisher’s bait, they can use the information to create fake accounts in your name, ruin your credit, and steal your money or even your identity.† This is a serious issue that takes some skill to recognize to evade taking the phisher’s bait. First and foremost is to educate yourself on phishing attacks or scams. Know what it looks like. There are many resources that you have available to you on the internet to learn how to spot a phishing attack and a legitimist email. The Department of Defense offer s a great course on how to spot a phishing email at http://iase.disa.mil/eta/phishing/Phishing/launchPage.htm. A more specific attack is whaling. Blogs.iss.net explains â€Å"The adoption of the term ‘Whaling’ within phishing is fairly new and may have been derived from the use of ‘Whales’ within gambling to refer to big-time gamblers and high rollers, but most likely come from the colloquialism for â€Å"big fish†. Regardless, Whaling describes the most focused type of phishing currently encountered by businesses or government – targeted attacks against groups of high-level executives within a single organization, or executive positions common to multiple organizations (e.g. the CTO or CFO). In a whaling attack, the phisher focuses upon a very small group of senior personnel within an organization and tries to steal their credentials – preferably through the installation of malware that provides back-door functionality and key logging. By focusing upon this small group, the phisher can invest more time in the attack and finely tune his message to achieve the highest likelihood of success. Note that these messages need not be limited to email. Some scams have relied upon regular postage systems to deliver infected media – for example, a CD supposedly containing evaluation software from a known supplier to the CIO, but containing a hidden malware installer.† So if you’re not a high level employee, you’re probably wondering how this is important to you. How phishers go about their scam is by obtaining little bits of what some would consider being harmless information from other employees about level executives and projects going on in the company. They start at the bottom to get more information at the top. So in all actuality it very much concerns everyone in the company. This leads me in to my last but important area of cyber security: insider threat. Frankly, that is you. Every employee is potentially an insider threat. So let me define it a little better with some help from searchsecurity.techtarget.com. â€Å"An insider threat is a malicious hacker (also called a cracker or a black hat) who is an employee or officer of a business, institution, or agency. The term can also apply to an outside person who poses as an employee or officer by obtaining false credentials. The cracker obtains access to the computer systems or networks of the enterprise, and then conducts activities intended to cause harm to the enterprise. Insider threats are often disgruntled employees or ex-employees who believe that the business, institution, or agency has â€Å"done them wrong† and feel justified in gaining revenge. The malicious activity usually occurs in four steps or phases. First, the cracker gains entry to the system or network. Secondly, the cracker investigates the nature of the system or network in order to learn where the vulnerable points are and where the most damage can be caused with the least effort. Thirdly, the cracker sets up a workstation from which the nefarious activity can be conducted. Finally, the actual destructive activity takes place. The damage caused by an insider threat can take many forms, including the introduction of viruses, worms, or Trojan horses; the theft of information or corporate secrets; the theft of money; the corruption or deletion of data; the altering of data to produce inconvenience or false criminal evidence; and the theft of the identities of specific individua ls in the enterprise. Protection against the insider threat involves measures similar to those recommended for Internet users, such as the use of multiple spyware scanning programs, anti-virus programs, firewalls, and a rigorous data backup and archiving routine.† It could also be an employee giving away what you would consider to be harmless information like an email address, a project you are working on at work, how many people you work with, or even where you work and the location of the building. Even with some of the best antivirus software, a stellar knowledge of phishing, and an elite cyber security system, an insider threat can penetrate that because they most the time have the credentials to log it to the system and pull resources ligitimently so it would go unnoticed. So how can you help protect you network or your company’s network from an insider threat? Simply, don’t be one. Always be sure to only give information that the other person is on the same level to receive. Secondly is to educate yourself on insider threat prevention. There is a plethora of resources to do just that. A helpful one is http://www.ussecurityawareness.org/highres/insider-threat.html. Also watch for signs of curious activity with co workers. Report any evidence or suspicions to your supervisor. So in conclusion, prevention starts with you. Buy up to date antivirus software. avoid risky sites, educate yourself on what a phishing attack looks like, know what a legitimate email is, don’t be an insider threat and educate yourself about things to look for from other co workers. Cyber security is a serious threat and is a major issue that needs to be taken seriously. It is not just something for the IT department to worry about but starts with every one making sure to take the appropriate security measures to make the cyber world more secure.

How Elizabeth Bennet is used by Jane Austen in Pride and...

How Elizabeth Bennet is used by Jane Austen in Pride and Prejudice as a form of social protest In the society of Jane Austens day it was imperative that a woman married a suitable husband in order for them to live comfortably and improve the social standing and wealth of her family. Women in high society did not work and so it was vital that they married well. This situation is best summed up in the book by Jane Austen as we are given an insight as to the motives of Elizabeths sister Charlotte as she considers an offer of marriage, she writes Without thinking highly either of men or of matrimony, marriage had always been her object; it was the only honourable provision for well-educated young women of small fortune, and†¦show more content†¦She is like no other woman in the book or indeed of her time and by presenting this character Jane Austen is challenging the stereotypical woman of the day contrasting the typically obedient and loyal wife of the time with a woman who is strong willed and determined, something that would have shocked readers of the day. Within the book Mrs Bingley gives the reader an insight as to what was looked for in a potential wife all of which ironically Elizabeth does not agree with A woman must have a thorough knowledge of music, singing, drawing, dancing, and the modern languages, to deserve the word; and besides all this, she must posses a certain something in her air and manner of walking, the tone of her voice, her address and expressions, or the word will be but half deserved, all of which Elizabeth struggled to comply with. It is her liveliness and her opinionated manner that ultimately attracts the arrogant Darcy to her, and her belief that that she should not conform to the idea of marrying for money and social status as was the general belief of the day. She is reluctant to place marriage at the centre of her ambitions without any regard for feelings or circumstances. Despite being intent on marrying for love and not money, Elizabeths strong principals almost put her familys financial security and future at risk. The belief of the day was that if a man proposed andShow MoreRelatedPride And Prejudice By Jane Austen Essay1724 Words   |  7 PagesThe 18th century novel, Pride and Prejudice, by Jane Austen, is a fascinating book about a young woman’s struggle with family and love. Pride and Prejudice was originally published in 1813, but, the most common version of the story, and the one used for this research, is from the version published in 1892, still by only Jane Austen, though many other authors have contributed to this book over time. Austen often references the class system at the time, often noting one of the multiple heroine’s struggleRead MorePride And Prejudice By Jane Austen Essay1711 Words   |  7 Pageshe 18th century novel, Pride and Prejudice, by Jane Austen, is a fascinating book about a young woman’s struggl e with family and love. 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Unfortunately, Austen does not create a match for Elizabeth who is her equal in terms of characterization. Mr. Darcy, Elizabeths sometime adversary, beloved, and, finally, husband, is not so carefully crafted as she, for his character is somewhat undefinedRead MorePride And Prejudice By Jane Austen1467 Words   |  6 Pages Pride and Prejudice by Jane Austen is a classic novel that has remained relevant even years after its release. Its themes and symbols are understandable to even the most modern of reader. One of the many themes is sisterhood, something that is focused on constantly throughout the novel. Elizabeth Bennet, the protagonist of the novel, finds many of her decisions to be based upon the actions of her sisters. Making sisterhood a main driving force. Whether they are confiding in each other for marriageR ead MorePride And Prejudice By Jane Austen1384 Words   |  6 PagesW. 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Federal Bureau Of Investigation Agent Essay - 907 Words

Federal Bureau of Investigation (FBI Agent) Associate Dean once said â€Å"It just dispelled all of the bad things I’d ever heard about the FBI†¦ that the FBI was sneaky and out to get people.† The career of the FBI is a remarkable and exciting career, because of the endless possibility of helping the world get better in any kind of way . The research will describe the career of Federal Bureau of Investigation , what is required to become a Federal Bureau of Investigation agent a successful career and the impact this career has on has on society. The Federal Bureau of Investigation hires an agent to act as the lead investigators in the United States. The duty of an FBI is to guard the United States from Terrorists and Foreign intelligence threats, also to provide leadership and criminal justice services. The FBI has to develop effective work plans. They need to arrange resources to accomplish goals. The FBI has an investigative partnership with cops, sheriffs, etc. The FBI investigates spies, hackers, pedophiles, mobsters, gang leaders and serial killers, they investigate everything that can be a threat to the country or other people. FBI agent is able to carry firearms and most of the time are only called when they need to find a high profile criminal. Agents might sometimes run into dangerous situations. They have to travel extensively, they’re lifestyles are very sudden they can be on the go at any time. FBI agent are in charge of investigating more than 200 differentShow MoreRelatedFederal Bureau Of Investigation Agent Essay922 Words   |  4 Pagescareer, because of the endless possibility of helping the world get better in any kind of way . The research will describe the career of Federal Bureau of Investigation , what is required to become a Federal Bureau of Investigation agent a successful career and the impact this career has on has on society. 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