The Cell Adhesion Practical

The Cell Adhesion Practical The extracellular matrix consists of many polymeric proteins and polysaccharides that are assembled into an organised meshwork. The extracellular matrix in our own bodies can be found as connective tissues such as bone and tendon. In animals the extracellular matrix forms a structure called the basal lamina. Basal lamina is a thin, tough, flexible sheet of matrix molecules and is essential in underpinning all of the epithelia. The basal lamina separates the cells and epithelia from the surrounding connective tissue and forms a connection between them. A Basal lamina determines the cell polarity, promotes cell survival, proliferation, and serves as a direct route for cell migration. The basal lamina in the extracellular matrix of animals consists of two main classes of extracellular macromolecules: fibrous proteins known as glycoproteins and gylcosaminoglycans which are polysaccharide chains covalently linked to core proteins to form proteoglycans. The matrix applies powerful influenc es on the cells. These influences are applied mainly through transmembrane cell adhesion proteins that act as matrix receptors. These matrix receptors have a main role in epithelial cells and mediating their interactions with the basal lamina. Many types of molecules can function as matrix receptors. The main receptors on animal cells for binding most extracellular matrix proteins are the integrins. Integrins are a large family of heterodimeric proteins that exist as both alpha and beta heterodimers. Many matrix proteins in vertebrates have multiple integrins. 5 can bind to Laminin and 9 types of ÃŽ ² subunits and 24 of ÃŽ ± subunits are formed from human integrins. ÃŽ ²1 subunits form dimers with 12 ÃŽ ± subunits. These subunits are found on all vertebrates and an example of this integrin is the ÃŽ ±6ÃŽ ²1 Laminin receptor. Fibrinogen is bound by the ÃŽ ²3 integrins. These integrins are found on many cells such as blood platelets. Platelets interact with fibrinogen during blood clo tting. It is known for ÃŽ ±2ÃŽ ²1 to bind to collagens, ÃŽ ±5ÃŽ ²1 binds to fibronectin, and ÃŽ ±6ÃŽ ²1 binds to laminins. The binding of a matrix component to an integrin can send a message to the inner part of the cell, and this causes the interior part of the cell to send a signal back to the matrix. The extracellular matrix of connective tissues plays an important role in many muticellular organisms. The connective tissue in the matrix is constructed from glycosaminoglycan polysaccharides which form proteoglycans and fibrous proteins such as collagen. The collagen fibres strengthen and help organise the matrix together. The fibrous proteins give the matrix the flexibility. Collagens are secreted in large quantities by the connective tissue cells. A main feature of a collagen molecule is its triple helical structure. The fibrous proteins give the matrix strength and form structures to which the cells can be anchored using glycoproteins such as Laminin and fibronectin that have ma ny binding sites for integrins on the cell surface. Laminin is the first extracellular protein synthesised in a developing embryo. It is made up of three polypeptide chains held together by disulphide bonds. The extracellular matrix also contains noncollagen proteins with multiple domains and specific binding sites for receptors on the surface of cells. These proteins organise the matrix and help cells attach to the matrix. The extracellular matrix components are degraded by proteolytic enzymes called proteases that act close to the cells that produce them. These proteases belong to two general classes: matrix metalloproteases and serine proteases. Matrix metalloproteases depend on bound Ca2+ and Zn2+ for activity. The matrix metalloproteases and serine proteases cooperate to degrade the matrix proteins collagen, Laminin, and fibrinogen. The adhesion assay to be used in this experiment involves using trypsin. Trypsin is an enzyme used to remove the living C6 glioma cells from the confluent dish. Gilal cells are important cells in the brain. The experiment will use rat tumour cells. The media will be grown in calf serum. The Calf serum contains growth factors which are used for cells to divide and also contains the ECM components such as fibrinogen. Trypsin is dissolved in a solution that contains a divalent cation cheladtum. This will interfere with the calcium and magnesium cations that are important f or cell-cell interaction and cell matrix interactions. The cells will be put in suspension to see how they attach to different ECM components and then placed in the two top lanes of a 96 well dish. The first lane will contain Laminin, collagen I, and fibrinogen. The 2nd lane is the control. When matrix components are placed on well they will stick but not all stick this is when BSA is used in second lane. BSA is a protein added to other ECMs to block the sites that the 3 different ECM components havent stuck to. (1)(2)(3)(4)(8)(9) Cell adhesion assays are useful in many aspects of cell physiology and the mechanism of the adhesive response itself. Cell Adhesion Molecules (CAMs) are proteins located on the cell surface that are involved with the extracellular matrix (ECM) in the process called cell adhesion. Cell adhesion molecules have been identified from epithelial cells and studies demonstrate the role of extracellular matrix proteins, fibronectin and Laminin in cell attachment to matrix. These proteins contain specific domains which interact with other matrix components such as collagen. Cell adhesion is a complex process that involves molecular interactions and receptor ligand binding. Adhesion assays measure the contacts between a cell and extracellular adhesion proteins and also provide information about the cellular events. Cell adhesion assays are used to test the ability of a cell line to adhere to a subtrate(4)(5). The methods for measuring cell adhesion can be divided into two types. In the first type of methods, cell adhesion is analysed under static conditions. Static assays are used widely to assess the adhesion of different types of cells such as fibroblasts and epithelial cells. This assay describes the cells ability to adhere. This method is simple to perform and provides assessment of the adhesiveness of cells to a defined extracellular matrix substrate for example Laminin. However, static assay methods poorly stimulate adhesion that occurs in blood. Therefore, a second method is used to measure the cell adhesion using flow chambers. The use of flow chambers enables the researcher to stimulate blood flow and to reconstruct cell systems in the presence of shear. Flow chambers are available commercially and most commonly used to study leukocyte adhesion, either with endothelial cells or to substrates purified ligands. The aim of this practical is to perform a cell adhesion assay with 3 di fferent ECM components and to investigate the ability of extracellular components to adhere to c6 glioma cells. (4)(5) Results In the practical there was an experimental error this is due to mixing with serum. Therefore only B9-B12 was used along with other colleagues results. Trypsin is an enzyme used to remove the living C6 glioma cells from the confluent dish. Gilal cells are important cells in the brain. The experiment used rat tumour cells. Trypsin is a protease used to degrade the ECM proteins. Trypsin is dissolved in a solution that contains a divalent cation cheladtum. This will interfere with the calcium, magnesium cations that are important for cell-cell interaction and cell matrix interactions. Cells are put in suspension to see how cells attach to different ECM components. Discussion From the results it is shown a hierarchy of ECM components can be concluded. Matrix 1 concludes to be fibrinogen; this is because it is the least to adhere with C6 glioma cells. Asano et al,(2004) stated that C6 cells adhered a little more than twice as well to type I collagen as to Laminin this can be clearly shown on the bar chart as the two greater bars (matrix 2 and 3) indicating fibrinogen as the least. Results for both individual and model show C6 glioma cells adhered well to both type I collagen and laminin-1 but C6 glioma cells did not adhere well to fibrinogen. A reason to why the C6 glioma cells did not adhere could be that they are derived from glial cells in the rat brain. Cells derived from gilal cells are unlikely to adhere with fibrinogen and therefore unlikely to express integrins which interact with fibrinogen and this can be seen clearly in both model data and own data. A number of integrins require activation to bind their ligand and anchor the cell to the ECM. For example alphaIIb beta 3 integrin from platelets cannot bind to fibrinogen unless the platelets are activated by binding collagen or thrombin in a forming clot. The activation leads to changes in the integrin which are linked to the cytoskeleton remodelling. In both the model data and own matrix 3 shows to have the greatest absorbance and adherence. This is because collagen is the most abundant protein and most of the extracellular matrix is made up of collagen. Collagen shows the most adherences to the cells this is because collagen contains more integrin receptors than the other ECM components. Trypsin breaks down the ECM properties and therefore cells appear separated under the microscope. A reason to a higher adherence to collagen could be trypsin breaks down the collagen components faster than fibrinogen. A low adherence is shown for fibrinogen this is because fibrinogen in the brain is low. Glioma cells originate from the brain and therefore they would not encounter with fibri nogen so the integrins only bind to collagen and Laminin.(3)(2) Another experiment in determining which integrins are important in cell adhesion to the ECM component is using antibodies. Antibodies identify the functions of the ECM receptors in cell attachment and cell migration by attaching to the alpha and beta and are also used to block adhesion to ECM .Blocking the function of the beta subunit inhibited cell adhesion to Laminin and fibronectin. These results were expected because a beta polypeptide can associate with several alphas to form receptors for collagen and Laminin. In the late 80s functionally blocking antibodies were used on ÃŽ ±2 integrins to block adhesion to collagens and ÃŽ ²1 integrins blocked the adhesion to several ligands. 2 Rat monoclonal antibodies have been used against integrins. Monoclonal antibody (mAb) 13 recognizes the integrin class 1 beta polypeptide and monoclonal antibody 16 recognizes the fibronectin receptor. These monoclonal antibodies were used to test the inhibitory activities in cell adhesion, spreading, and migration using w138 human fibroblasts. It was shown mAb 13 inhibited the attachment and spreading of w138 cells on fibronectin and Laminin substrates. It was shown Laminin mediated adhesion and was sensitive to mAb 13 compared to mAb 16 which showed cell attachment to fibronectin but not Laminin. This shows this receptor is not involved in Laminin cell adhesion. (7) The ÃŽ ±1ÃŽ ²1 integrin can bind to collagen types I, II, III, and V but favours other subtypes such as type IV and XIII. This is why ÃŽ ±2ÃŽ ²1 integrin is the major receptor for type 1 collagen. An experiment in determining the collagen receptors is to see the effect of collagenase-cleavage of type I collagen on a2b1 integrin-mediated. Collagenase-3 cleavage of type 1 collagen has shown to have an effect on ÃŽ ±2ÃŽ ²1 integrin. An isolated ÃŽ ±2ÃŽ ²1 integrin and ÃŽ ±2 integrin A- domain were found to bind native collagen and native k fragment. It was shown integrins were lost after heat denaturation of the collagen fragments and shown human fibrosarcoma cells (HT1080) adhered to type 1 collagen and k fragment at 37Â °C. HT1080 cells cultured on type 1 collagen and collagen fragments expressed high in ÃŽ ±2ÃŽ ²1 integrin on cell surface and very little to ÃŽ ±vÃŽ ²3 integrin detected by immunolocalization. This experiment shows that ÃŽ ±2ÃŽ ²1 integrin binding to type 1 collagen i s dependent on the maintenance of the collagen triple helical conformation. (6) Inconclusion integrins are important adhesion molecules involved in cell-cell- matrix adhesions.

Custom Coffee & Chocolate Essay

According to the overview of Custom Coffee & Chocolate’s business, Bonnie Brewer and Stacy Kim run a small cafà © where they serve unique coffee, chocolates and maintain a shop. The cafà © is located near a university, where students, faculty, staff, and local residents are the main customers. They have run the business successfully, working together without any employees and selling their products only in the cafà ©. They have started to think about further development. The main question at this point is what strategies they should launch to achieve that aim. The first step to answer on that question is analyzing the company’s internal environment and identifying it strengths and weaknesses. The greatest strength is undoubtedly knowledge acquired by the owners during business and marketing courses and there experience gained at work. The owners also chose an interesting location with many students who they know and understand very well, which combined with the offer is strength of their business. Customers can sit in comfortable chairs, on couches, and at coffee tables, and chat or read newspapers between classes. As we consider weaknesses, we can identify a few problematic things. The largest of these is the limited access to services due to the small space in the cafà ©. Moreover, lack of employees makes it impossible for the cafà © to open longer, because the owners also have to handle the finances. There is also no possibility for customers to order goods over the phone or online, and for the cafà © to deliver service to locations on campus, such as dorms and lounges where students are studying. Another weakness is quite poor menu, which should be extended by new products. Additionally, too little attention is paid to social inclusion by organizing events such as poetry readings or discussion groups, and conducting a blog which could integrate students around the cafà ©. Lack of a wireless connection that customers could use is a significant drawback. The last weakness, which is really important, is a lack of financial reserve. The owners exhausted their savings and the initial small business loan, and  their six-month lease is up. The second step is analyzing the company’s external environment and identifying it opportunities and threats. The greatest opportunity for the development of the company is the ability to reach a wider range of customers who are potentially interested in using the service of the cafà ©. Custom Coffee & Chocolate’s business plan bases on purchasing only fair trade coffee and chocolates made by a few local suppliers. It is worth considering the possibility of obtaining a better price from producer in exchange for promoting their brands. It is also possible to extent the number of potential suppliers, which is likely to enable the cafà © to purchase products at much better prices. An opportunity for the development of the company is the application of new technologies such as online shops, websites or blogs, to allow better contacts with customers on the Internet. Unfortunately, threats also can appear in the company’s external environment. It could be a decrease in the number of students at the University or changed location of classes. The biggest threat may be the emergence of competitors offering similar services at lower prices or better quality. The strategy adopted by the company, which consists of buying products from local suppliers and ordering only fair trade coffee, may cause a risk of introduction of cheaper substitutes on the market by competitors. The last but not the least threat is connected with the need to increase the cafà ©Ã¢â‚¬â„¢s debt by taking additional loans for further development. After determining the strengths and weaknesses of the company’s internal environment as well as the opportunities and threats of the company’s external environment, it is necessary to propose some strategies that take into account the interdependencies between particular elements of SWOT analysis, which will help to develop the company in the right way. One of the ideas is to acquire new customers by increasing the availability of products. Arranging a sale of products through the online store and allowing customers to play orders by phone, organizing appropriate marketing campaigns on the website or in local media, gives opportunities to achieve that goal. Ordered goods would be delivered by courier. At the initial stage, the range of delivery might be limited to locations on campus, such as dorms and lounges where students are studying. This strategy would be built on opportunities of using the Internet and growing the customer’s base without the necessity of looking for larger premises to run  a cafà ©. It is necessary to hire employees to allow the owners to focus on strategic management activities in the cafà ©. It would give Bonnie Brewer and Stacy Kim opportunity to use their knowledge and experience and take over more of the finance and marketing activities. Another interesting strategy that can protect the company against the loss of customers is building social cohesion by turning the cafà © into a cult meeting place for students, faculty and other staff of the University. Organizing poetry readings or discussion groups could be a great idea. Moreover, those activities would be supported by a blog which would be moderated by the cafà ©Ã¢â‚¬â„¢s owners. This way, drinking coffee or eating delicious chocolates would be only a natural addition and benefit during meetings. In that case, the main strength is the specific location with students, faculty and staff whose the owners of the cafe know and understand well. Having loyal customer groups may be particularly important when some competitors appear on the market. This way, the company could significantly reduce this kind of threat. A very effective strategy which could be used is building a recognizable brand of their own products. They should be presented as unique and not available from other competitors. In addition, it might be possible to prepare a wider range of products which could be better suited to the local customers needs. This strategy should be based on the promotion of local products and suppliers of products and high quality fair trade coffee. It could be an opportunity to get additional money from sponsors of the cultural and community building mission carried out by the owners of the cafà ©. The above strategy is based on the strength connected with high quality products which take a niche place on the market and cannot be easily replaced. It protects the company from the threat of competition and improves its competitive advantage.

Since the dawn of civilization, people agonize for? Essay

Informatics provides the means for a development and growth of social networking services. The cost for communications has been reduced by enabling simultaneous transmission of multimedia content. On professional level, videoconference systems can accommodate people from all over the world to organize meetings and arrange their business, hence the issue of distance and time has been eliminated. On personal level, people can contact with their friends at anytime without considering the economical problem or the distance, especially when someone is resident of a foreign country. Also informatics , allows us to store, analyze and search through massive amounts of data on very short time. In addition, informatics links various devices such as smart-phones, tablets and personal computers. Modern devices are adjusted for usage of the same protocols and standards, which overcomes hardware difference. Moreover, Informatics has strong contribution on the development of algorithms, applying it on cryptography. Besides that it is used for computing, biometrics and primarily for graph algorithms, which solve problems related to scheduling, connection, transportation, games and network analysis. Algorithms are further associated with neural networks. That makes it even more efficient for data study, decision-making process and machine learning. As the complexity of solution grows and the number of people involved in the project increases, so does the further development complexity. Even there, Informatics responds with various solutions by software engineering and IT management. Neverless, the users don’t want a perfect workingly services, it also has to be interesting and handy for their needs. Therefore, informatics has to offer solutions and ideas on the field of user-user interaction, which is actually based on user-service interaction. Numerous social games or virtual worlds, where we can interact with other users like in the real world gives us the feeling that we communicate directly with other users, but such services have many tasks behind the scene. Users create groups and share interests, experience, feedbacks, habit. In the backend, the social networking service may collect, analyze and learn from data. For instance,  chat service can build database of words we use the most and add them in auto-complete functionality, which speeds up search and messaging. The main goal of user interfaces is simplified user-service (human-machine) interaction and therefore, better user experience. A social networking service has to be easy to use, attractive, competitive and scalable. It must treat users as spoiled, lazy and always unsatisfied. They want real-time monitoring, the feeling of a safe and effective application. The best systems for social networking are those, which make it possible for users to use it easily in spite of their age, previous computer skills or specific demands. Also those, who have special needs, such as people with visual, hearing or any other disability, should benefit from it. Since users have become more demanding, various browsers do not support same JavaScript Functions and HTML5 with CSS3 has brought new options, such as web storage, canvas 2D drawing, geo-location, responsive design etc. – frontend development became more complex. Static generated pages have been replaced by dynamically loaded content and scripts. We can choose from a variety of open-source JavaScript frameworks that differ in purpose, speed, assertion, community size etc. Smart devices make the user experience even more diverse and exciting by using sounds, vibrations, digital camera and other built in hardware. 3. Social networking services threats Social networking services as a product of Informatics and social networks gave us unimaginable opportunity to connect with people. On the other hand, they have many disadvantages. The illusion of safety, often make us forget the possible consequences. Heedless of the danger, we share our personal data with people who we do not know, therefore a lot of misuses appear. Companies trade information about users, there are many cases of identity thefts, in brief, social networks offer a chance for various criminal offenses. We know very little about how and where they store our personal data and who has access to them. Even when we adjust our settings for completely private accounts, our friends or services may inadvertently leak our personal information. We do not know whether they will ever actually be  deleted. Immense amount of users is not only bullied interest in networking, but also invaders, who attack and abuse either from personal satisfaction, or even receive payment for their work. They use various attack methods, based on personal profiles, trust or exploits, e.g. social networking worms, such as Koobface. 4. Influences Similar to Newton’s third law of motion, one can conclude that everything society receives, passes on in the environment. The impact of the social networks on society indirectly affects other fields like economy (PayPal Inc. 2013), education, politics (Shirky 2011), entertainment etc. These fields see social networks as an opportunity to approach to the crowd, mostly for the purpose of manipulation (Benham, Edwards, Fractenberg, Gordon-Murnane, Hetherington, Liptak & Mintz 2012). The most common use of manipulation can be seen in setting imaginary guidelines, viral marketing and targeted advertising (Mitra & Baid 2009, 366-372). Referring to paragraph three, companies have free access to some of our personal data, or they can reach us by low cost targeted advertising, offered by social networking services (Eddy2012). It may be based on specific personal profiles, such as sex, interests, education, status, age, geographical location and even on private messages content.Motivated by Domingos and Richardson Viral Marketing Model (Richardson & Domingos 2002, 61-70), numerous companies are searching for cheapest ways to adopt a new product or innovation, whilst seeking for smallest subset of individuals, which may trigger a large cascade of further adoptions. They are interested in individuals who have the greatest impact on the target social network (Kempe, Kleinberg & Tardos 2003, 137-146). Moreover, the use of technologies offered by social networking services can reduce the development costs (Churchill & Halverson 2005) or can simplify the use of applications e.g. registration system with the existing user accounts from social networks. Furthermore, with clicking on the ‘’like’’ buttons one contributes to profiling users (Hung, Huang, Hsu & Wu 2008), which is especially  interesting and useful in research, determining trends, positioning in the search engines etc. Recent studies, based on a dataset of over 58,000 volunteers suggest, that easily accessible digital records of users behavior in social networking systems, can be used to automatically and accurately predict a range of highly sensitive personal attributes (Kosinski, Stillwell & Graepel 2013), e.g. religious and political views, intelligence, happiness, parental separation etc. However, the economy influences social networks by adapting social networking and its services according to the needs of the users. Economy’s investments in the most prospective social networking services have a significant influence as they allow further development, starting new projects, research and changing habits of users. 5. Trends The expansion of new research areas in the field of Informatics led to re-implementation of social interactions in social networking services. Therefore, changes in the field of social networks are incredibly fast to keep up with. Nevertheless, in few last years, we may notice two main trends that have emerged; visualization of content and mobile use and sharing (Bullas 2012). For trend of content visualization we can find the reasons in exceptional progress in technologies for detecting patterns in digital images. Thus allow analysis and image processing on servers where social networking services can get even more personal information about users. This motivated investments in deeper research, and in further years we may expect DNA data sharing, through which social networking services may link us and connect to all our near and distant relatives. Mobile use and sharing trend will continue to grow, since use of smart mobile devices is increasing. Better user experience, mobility, availability, as well as faster and cheaper mobile data transfer are just few of many reasons. Recent technologies, like digital glasses (Google Inc.2013), may allow us to record and share each moment of our life. Despite strong efforts of the health sector, where currently more than 350 million people suffer from addiction of social networking services (Bullas 2012), at the continued growth of those services trend is likely to continue. 6. Conclusion Only a few decades ago, we have not even intuited how important role will Informatics have in our daily life and how huge contribution will have through its social networking services on us, social networks and global economy. Social networking services as part of social networks are one of the hottest research topics nowadays. We have poor insights in their long term impact on society, since we have witnessed true expansion only in the last few years. However, the first negative implications, like privacy issues are already visible, as well as first solutions, which aim to limit misuse. Recently, Apple has published a patent, which targets social network stalkers and spammers. The excessive use of social networking services promote narcissism (Buffardi & Campbell 2008, 1303-1314). As one has thousands of virtual friends, this could mean that in real life he does not have them at all (Tong, Van Der Heide, Langwell & Walther 2008, 531-549). And finally, one of the studies suggests tha t frequent use of social networking services may lead to asocial user behavior (Orr, Sisic, Ross, Simmering & Arseneault 2009, 337-340). These are just some of the findings of studies pointing to major changes in society.

An Urgent Issue in Our Society - Violence Against Women

The concept of violence might only represent a negative image. It refers to a set of radical human behaviors, for example, perpetrator would use physical force to hurt or kill people on purpose. Besides, the typology of violence in World Health Organization (WHO) has an explicit structure, which comprise physical, psychological, and sexual to families, partners, friends or strangers (). However, in this essay I shall focus on critically analyzing Farkhanda Younis’ story, which could reflect how violence against women as an urgent issue in our society. On 19 April 2013, a six-year old boy called 999 said he could not assess in his mother’s room because it was locked. After police came that they found a young mother’s body.†¦show more content†¦In other words, domestic violence could be a way to dominate or control women by men. Moreover, females might usually play a more careful, affectionate, or emotional role in the family. Hence, in the men’s subconscious that they might have illusions, which their wives could tolerate everything include personal harm. However, they forgot family is mutual respect and violence could not solve any problems. Thus, based on psychologists xx opinions that Dobash and Dobash’s theory might relieve of charges of discrimination. Furthermore, xx emphasizes if we use social justice to maintain social stability that domestic violence might be the biggest destroyer. Firstly, the violence themselves could be seen as a potential dangerous group, which might harm others, such as acquaintances or strangers. In addition, the domestic violence also could be seen as a dangerous element, which might break the ethical guideline. Without doubt that Ms Younis’ case awoke more and more people know domestic violence would no longer just harm or abuse women that death also became a horrible result of violence. As Xx states, a good prevention might be a better way to reduce or solve problems. 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