Participles Fused and Otherwise

Participles Fused and Otherwise Participles Fused and Otherwise Participles Fused and Otherwise By Maeve Maddox On the off chance that you dont recognize what an intertwined participle is, read on. The current participle is the type of the English action word that closes in ing: walk strolled strolling To work as an action word, the current participle must be utilized with an assistant action word: Jack is fixing the rooftop. Utilized without an assistant action word, the participle holds some shadow of its verbal root, however works as different grammatical features. participle working as descriptive word: Mr. Jones is a caring spouse. participle presenting participial expression: Sitting by the window, I watched the motorcade. (The expression is descriptive, depicting I) participle working as a thing: The ing participle structure can likewise be utilized as a thing. All things considered it gets another name and is known as an ing word. Ing words Planting is my preferred leisure activity. (thing, subject of is) He enjoys shooting skeet. (thing, object of preferences) He wants to discuss chasing. (thing, object of the relational word about) Do you mind my posing an inquiry? (thing, object of psyche) NOTE on Example 4: If I had composed Do you mind me posing an inquiry, a considerable lot of my perusers would rush to reprimand me for having composed a sentence containing a melded participle. Combined participles The term combined participle is credited to H.W. Fowler, who loathed them. Heres the definition from the OED: combined participle a participle viewed as being joined syntactically with a former thing or pronoun, as opposed to as an ing word that requires the possessive, or as a conventional participle qualifying the thing. The combined participle lives in a similar classification as the split infinitive: a few essayists severely dislike it and will evade it any expense, while others perceive that, occasionally, defusing an intertwined participle is more awful than disregarding it. My training is to utilize a possessive thing or pronoun before an ing word in a sentence like the one above. In the event that the outcome is terrible or absurd, I make sense of how to revamp the sentence without utilizing the - ing word. Talking is another issue. In discussion I most likely breaker participles everywhere. A few perspectives with respect to the combined participle: H. W. Fowler David Rusinoff (click on intertwined participle in the casing) American Heritage Book of English Usage Columbia Guide to Standard American English William Safire Need to improve your English in a short time a day? Get a membership and begin accepting our composing tips and activities day by day! Continue learning! Peruse the Grammar class, check our well known posts, or pick a related post below:Passed versus PastCapitalization Rules for Names of Historical Periods and MovementsLetter Writing 101

Is Victor Frankenstein the Modern Prometheus?

Is Victor Frankenstein the advanced Prometheus? Mary Shelley’s reason in the caption of her book, Frankenstein; or, the cutting edge Prometheus is to think about Victor Frankenstein and Prometheus, child of Zeus. By indicating numerous likenesses between the two, she has strong thinking that Dr. Frankenstein is, from multiple points of view, the â€Å"modern Prometheus. † Although both primary characters make a difficult they have no power over, they gain proficiency with their exercise of altering the law of nature. It requires some investment, discipline, and tests for the them two to comprehend their bad behaviors. Shelley’s book shows numerous likenesses among Victor and Prometheus.In the starting we are persuaded that Victor is a researcher who, through investigation and examination, can re-make life. By utilizing transformation he utilizes old dead body parts to make a larger than usual and abnormal animal (Shelley, standard. 91). Like Prometheus, Frankenst ein made something he had no force or power over. Prometheus took fire from the divine beings and offered it to people, once as well as twice. In doing as such, the two of them conflicted with all laws of nature. While Victor Frankenstein made a nonhuman animal and Prometheus took fire from the divine beings they were both rebuffed very severely.After Dr. Frankenstein breathed life into his animal, out of disturb, he fled. â€Å"How would i be able to portray my feelings at this calamity, or how outline the blackguard whom with such interminable agonies and care I had tried to frame? † (Shelley, standard. 121). Frankenstein couldn't stand seeing his creation. Seeing it made him squeamish and sick. The monster’s curiously large body and unattractive extents brought Victor much desolation. The animal, feeling surrendered, went to seethe and assaulted Victor’s friends and family. His first casualty was William Frankenstein, Victor’s dearest brother.Giving Vi ctor brief period to fix from his heartbreaking misfortune, the animal assaulted once more. This time he lost his closest companion, Henry Clerval. During these disastrous occasions, Victor realized this was his issue. He made a beast who might keep harming him by ending the life of the individuals he thought generally about. The last passing of Victor’s family was his sweetheart spouse, Elizabeth. The animal murdered her out of vengeance since Frankenstein would not complete the female buddy he requested to be made. Frankenstein lost the individuals nearest to him while Prometheus was rebuffed in a totally different yet, equivalent way.Prometheus felt that people ought to be better than the divine beings. Having more force and the capacity to make apparatuses and weapons, he gave them shoot. Zeus was angry about this, seeing he had removed fire from them previously. At the point when Prometheus offered fire to man the first run through, Zeus was irritated with outrage and di sclosed to Prometheus that fire was not a force man should hold. Challenging Zeus, Prometheus gave it back to man in any case. This was not a blessing Prometheus could give to people after it was removed once. Later rebuffed for his activities, he was condemned to be fastened to a stone with the organization of a hawk, hungry for his liver.The anxious flying creature picked at Prometheus’ liver every day as the sun would disseminate. Overnight his liver would develop back however he would bear this horrifying and agonizing agony once more. The two men experienced severe discipline. Prometheus was tormented truly, while Victor was harmed on an enthusiastic level. After their discipline, both Dr. Frankenstein and Prometheus were offered chances to substantiate themselves. The animal went to Victor and requested that he make a female partner. In the wake of perceiving how merciful the individuals he watched were, the animal needed somebody to impart those sentiments to. I reques t an animal of another sex, however as frightful as myself†¦Ã¢â‚¬  (â€Å"National library of,† 1998). Dr. Frankenstein started amassing the body parts to fabricate a female animal. It took him months, yet before he completed he ruled against it. He considered how he was unable to control the animal he previously constructed. The idea of there being two of them out there on the planet sickened him. Victor was tried by having the chance to make a subsequent beast. By declining to do as such, he at long last discovered that attempting to play God isn't a job to be taken lightly.After Dr. Frankenstein denied the animal a mate, he before long understood this was something worth being thankful for. Prometheus was tried along these lines. His dad, Zeus, gave Prometheus two choices. As there was a prediction that an offspring of Zeus would one day oust him, Prometheus, who had the endowment of foreknowledge, could reveal to Zeus the mother of the youngster who might do this to him. His subsequent choice was placed into two sections, there must be an everlasting that was happy to kick the bucket for him, just as a human who could slaughter the bird and break Prometheus’ chains.Chiron, the centaur who prepared Hercules, was unfading and offered to surrender his life for Prometheus. Hercules executed the bird and broke the chains (â€Å"The creation of,†). Shelley utilized the similarity between Victor Frankenstein and Prometheus in her book. By indicating how these characters are indistinguishable in various perspectives, she exhibited how Dr. Frankenstein is without a doubt, the cutting edge Prometheus. The two men assumed the job of playing God by making issues they couldn't oversee. Considerably more thus, the two of them endeavored something no one but God could decide or have power over.While Victor Frankenstein made a beast out of dead body parts, Prometheus resisted the Gods by giving man the intensity of fire. Every one of them, being rebuffed for doing as such, scholarly their exercise. Shelley’s character of Victor has comparative qualities as Prometheus. They were both extremely brave and nervy. The two men had the desire to make; however it didn't work in support of them it outlined their resemblance. Mary Shelley depicts Victor as a man who conflicted with God and the law of nature. Like Prometheus, Frankenstein was very tenacious and sadly for the two men it was past the point where it is possible to fix the harm they had committed.Dr. Victor Frankenstein is inconclusively, the advanced Prometheus. Reference Page Shelley, M. (1818). Frankenstein; or, the cutting edge Prometheus. London, England: Lackington, Hughes, Harding, Mavor, and Jones Publishing House. Recovered from http://www. gutenberg. organization/documents/41445/41445-h/41445-h. htm The making of man by prometheus. (n. d. ). Recovered from http://www. greekmythology. com/Myths/The_Myths/Creation_of_Man_by_Prometheus/creation_of_man_by_pr ometheus. html National library of medication. (1998, February 13). Recovered from http://www. nlm. nih. gov/frankenstein/prelude. html

Surprisingly enjoyable experiences

Surprisingly enjoyable experiences Power tools East Campus brings to mind rollercoasters and huge construction projects. Fanciful monsters built up from wood and screws. Less so the things that are built, but more the ambition behind it, more the fact that it’s  possible, that you didn’t need a company and dozens of weeks, that a couple of undergrads working together could cobble one up. This year, for REX, one of the things being built was a fort.  No experience required, was the advertisement. Come and help out. The night I moved in there was only a hexagonal frame in the courtyard, and by the next afternoon they’ve put pillars and more stuff in the hexagonal frame and they looked ready to start the next layer. The plans say it would be  three storeys high. Imagine that:  three storeys. And the deadline was in a couple days. Part of me wanted to help, but part of me was scared. Sure, they said no experience required, but I still couldn’t bring myself to ask. The whole project just felt like something so huge and so untouchable, something so carefully and meticulously thought out, something for actual engineers, and not the unexperienced prefrosh I was. It felt like a ridiculously complicated task.And maybe another reason was just that power tools are scary, that drills and saws are grown-up tools, a thought drilled into my head when I was twelve, that it wasn’t safe, that it was  scary. And maybe another reason is that trying new things out, in general, is just scary, because how do you talk to people? How do you ask for help? What if I make a mistake? I don’t know why I tried doing it anyway. But there were five of us prefrosh who all wanted to try it and didn’t know how to help, and there was an upperclassman who was willing to teach us. It was far less scary and far less complicated than it was in my head. I mean, you put on a hard hat and wear glasses. To use a power saw, you align the wood, press a button, push it down, and it chops off the wood you need. To use a drill, you grab a screw, align it, press a button, push it down, and it drills the screw in. Do these a couple times and you have a railing. After a little training, it really wasn’t that hard. It’s meditative, even. The actions are repetitive, but fun. I came back the next day to help out, and the next day to help out some more. Despite using drills and screws and saws, despite loud noises and sawdust flying in the air and getting all over your shirt, I found it oddly relaxing. Live action role-playing I’ve always wanted to try out live action role-playing. Sure, there’s this stereotype that it’s a bunch of nerds who dress up and fight each other with foam swords. But it sounded like a lot of fun, like being the actor in a movie you don’t know the plot of. I was pretty excited to find out about the Assassin’s Guild, a live action role-playing group here, and I signed up for their first game, which was held last weekend. It’s set in the universe of Isaac Asimov’s robot novels. Some people’s roles were humans, and others were robots, and the robots generally followed the Three Laws of Robotics. The game space was pretty big, spanning three floors of five buildings01 buildings 24, 26, 34, 36, 38 . Part of my role involved confronting people, terse negotiation, and making compromises in order to achieve my character’s goals. There’s something about asking to talk with someone privately, making wild accusations, and getting information, which is pretty fun. Another part of my role was looking for items I wanted, which were in packets taped to the walls of the game space. Which meant that I searched all three floors of all five buildings. This was time-consuming. In order to actually acquire items, you’d have to do this “hacking” mechanic, which involved shuffling a deck of cards seven times and then laying them out. This was also pretty time-consuming. Towards the end, people got suspicious of me and decided to restrain me. The whole interaction looked something like this: a group of five people approach me. guy: Knock Out 2. other guy: Assist 2. guy: Knock Out 4. other other guy: Assisâ€" me: okay okay i get knocked out i sit on the floor. It was so fast I didn’t even see who they were. They then decided to search all of my items, which consisted of me handing them the slips of paper that were in my pockets. They also took my energy blaster, which consisted of me handing them my Nerf gun and my foam bullets. It was pretty fun. I still don’t understand everything that happened, but it was surprisingly fun, and I’d love to do more in the future. Square dancing A misleading name, because it’s less like dancing and more like trying to remember what a Star Thru is after doing a California Twirl and having to remember that you and your partner are called centers, and after changing location you guys are now called heads and then a Four Ladies Chain is called and you’re moving to another spot, and then the caller asks for the sides to promenade, but even if you and your partner are heads you have to move forward in order to make way for them, and then you’re asked to partner trade and you have to remember to pass right-to-right shoulder, and suddenly everyone’s clapping because an Allemande Left was called and you quickly go to position. If you don’t understand anything written above, that’s okay. It would still be representative of what it feels like. I got dragged into square dancing when one of the upperclassmen living on our floor invited me to go to the Tech Squares’s02 MIT’s square dancing club intro night. I had a vague idea of what square dancing was. The caller gives commands, and people move around according to these commands, but I didn’t really know anything else. Again,   no experience required, and it really was no experience required. By the end of the night I could follow along to a handful of basic calls, and I enjoyed it enough to sign up for square dancing as my PE class this quarter. 21W.022 That is the class number for  Writing and Experience: Reading and Writing Autobiography. It’s a CI-HW, which means that it’s a class that involves a lot of writing. The discussions we have in class are surprisingly lively. It’s completely unlike any of the humanities classes I had back in high school. Everyone’s contributing their ideas about the text, and it feels more like a discussion between us students, rather than a question-and-answer session led by the professor. I also find the topics interesting in a personal sense, since writing about myself through blogging has been an important part of my life, and the class has gotten me thinking about these things. 15.000 And  that is the class number for Explorations in Management. Its a class that meets from 7 to 9 on Monday nights. It’s a discovery class, so it only carries three units, and there’s not a lot of work. In our first meeting, Professor Andrew Lo gave a talk about how financial engineering can help fund biomedical research. And there were several  literally hair-raising moments during the talk. It was mind-blowing. I enjoyed that first meeting more than any of my other, more technical classes, which is really surprising to me, because I don’t know anything about Course 15, and only signed up for the class on a whim. I’m really looking forward to the rest of the class. And I could go on. I could tell you about how I went to a estimathon hosted by Traders at MIT, and had fun arguing with some people I just met about how many bus stops the MBTA has, or how much volume the Antarctic Ice Sheet had, and how the time went by surprisingly quickly. Or about the ESP worksession I went to on Wednesday night, where I helped diagnose some email problems, which was surprisingly fun. If I were to distill this whole blog post into a single line of advice, it would be this: try new things. The last few weeks have been a constant string of surprises, of things I never thought I would like, but enjoy a lot. It’s not the case that I enjoyed every new thing I tried. Auditioning for the Asian Dance Team made me realize that I didn’t enjoy learning dance routines. Going to some more Traders at MIT events made me realize that I didn’t really want to join their group, or at least, not now. Playing some games with the MIT Poker Club made me realize that I didn’t enjoy poker that much. Why try new things if there’s a chance you won’t enjoy them? I guess it’s because of the small chance that you will  enjoy them. If you try ten things, end up disliking nine of them, but liked the last one enough that you do it a hundred times, then it would have been worth it, wouldn’t it? There’s so many new experiences that MIT is offering, and I love how I can use this first semester to explore them. Post Tagged #help writing is hard buildings 24, 26, 34, 36, 38 back to text ? MIT’s square dancing club back to text ?

Top Notch Tech Essay - 742 Words

Landing a Top-Notch Tech Internship One of my main goals this year is to land a top-notch internship with a technology company. I have been trying to read as much as possible on the best ways to accomplish this. There are many books and articles that have helped me, offering advice on interviews and what sort of skills I need to develop to obtain one of these internships. An article that I read â€Å"How to land a top-notch tech internship — and a tech job — while you’re still in school† by Michael Deng, gave some great input and I think it impacted my sense of professional identity and pertains to my goals that I have set this year. The article opens by talking about how each student needs a strong foundation. Without core math and†¦show more content†¦You should highlight your projects and past experiences (internships/jobs) with summaries and get rid of irrelevant details that only take up space on the resume. After completing your resume, have peers and teachers review it to make sure that it makes sense to future employers. Finally applying and interviewing are the final pieces for internships. One great way to get recruited for an internship is by emailing the recruiter directly. The article stated that LinkedIn is a great way to get the email of the recruiter. Follow up the email in week if there is no response, keep it concise and attach your resume. After contacting the company, an interview will most likely be set up next. There are many books that are recommended to get an idea of what might be asked. The article advised reading â€Å"Cracking the coding interview† which will help prepare programmers for technical interviews. Mock interviews are a great way to practice your responses to get an idea of what you want to say. Deng explained there are 4 stages for most technology companies, the coding challenge, recruiter chat, phone interview, and onsite interview. 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How to Calculate an Equilibrium Equation in Economics

Corporate Culture and Its Influence on the Development Free Essays

â€Å"CORPORATE CULTURE AND ITS INFLUENCES ON THE DEVELOPMENT OF MANAGEMENT CONSULTANCY AND BUSINESS SOLUTION JOINT STOCK COMPANY† INTRODUCTION ************ 1. Rationale of the study The term â€Å"Corporate culture† has appeared in the world and become a hot topic of many researches since the late 20th century. When the business environment faces a number of unpredictable changes and challenges resulting from integrity process and economic crisis, enterprises from any size and any country must recognize the important role of corporate culture to their survival. We will write a custom essay sample on Corporate Culture and Its Influence on the Development or any similar topic only for you Order Now However, corporate culture is truly concerned only in big groups and corporations and some medium-size companies. In Vietnam, the number of enterprises that have a proper strategy to develop their own culture is nearly insignificant. It is the fact that each company has its own culture may be without recognizing and controlling it. In the simplest way we can understand corporate culture as the environment within a company in which all staff and workers communicate and work together. Hence, if we pay a certain attention, we can easily see that corporate culture has a strong affect on the cooperation between all employees, the productivity in particular and the development of the company in general. Moreover, in consulting companies, the reputation plays a key role in the company development for it influences the number of customers trusting in and using their consulting service. However, the reputation of a company is contributed from the professionalism that is reflected through the working environment or, in other word, the corporate culture of the company. Therefore, it is necessary to study corporate culture and its influences in real consulting companies in Vietnam like Management consultancy and business solution Joint stock Company. In conclusion, such researches about corporate culture like the one being conducting in this study somehow will be highly appreciated by businesses. Especially in 2013, the economic outlook is considered to be worse than that in 2012. That is the reason why each company has to find out a developing strategy which can make them outstanding from others. Therefore I would like to choose â€Å"Corporate culture and its influences on the development of Management consultancy and business solution Joint stock Company† as my thesis in this study for its huge value and meaning to our economy. 2. Aims of the study The primary aim of this research is to study about the corporate culture of MCBS JSC and its impacts on the development of the company. Then the author will give some recommendations with the hope that they can help MCBS JSC find out their comprehensive strategy for further development. 3. Scope of the study Because of the limitation of time, this research will not cover all components of the corporate culture. It will just analyze some important and prominent factors which have significant effects (both positive and negative) on the development of MCBS JSC. More importantly, the research does not look at the corporate culture of MCBS JSC from the foundation up to now but in the two recent years (in 2011 and in 2012) for the reason that when the economic shows a sign of downturn corporate culture becomes more and more meaningful to the lifespan of its owner. 4. Methodology Data analysis is the most frequently applied method in the study. Besides, the author also conducts surveys by using questionnaires in order to get more information. In concrete, in the first stage lasting two weeks, the researcher collects data from various sources: media, employees and internal files†¦Then, the researcher carries out some surveys to have more specific and precise information in two later weeks. Lastly, in the four following weeks, data analysis method will be adopted to produce final conclusions. 5. Organization of the study The study is divided into three parts: The first part is the introduction dealing with the rationale, aims, scope, method, organization and significance of the study. The second part is the main part with three chapters: * Chapter 1 is the literature review which provides readers a general review of what corporate culture is, its components, and how it affects the company development. * Chapter 2 refers to the research methodologies. This means that it will describes in detail sources of data, methods applied in the research and their progresses as well as their outputs. * Chapter 3 is the main part of the study which consists of some discussions. This chapter will point out strengths and weaknesses of the current corporate culture in MCBS JSC. Then, the author will suggest some actions that could be done in MCBS JSC in order for them to develop a proper corporate culture. The last part is the conclusion that summarizes the content of the paper and gives some suggestions for further study. 6. Significance It is hoped that this research will be a helpful reference paper for the management of MCBS JSC in specific and for businesses in general to build up their most appropriate corporate culture. Table of Contents INTRODUCTION 1. Rationale of the study1 2. Aims of the study2 3. Scope of the study2 4. Methodology3 5. Organization of the study3 6. Significance4 CHAPTER 1: LITARATURE REVIEW 1. 1 Definition of corporate culture 1. 2 Components of corporate culture 1. 1. 1 Structure and roles 1. 1. 2 Espoused mission and goals 1. 1. 3 Rules, regulations and procedures 1. 1. 4 Formal codes of behavior 1. 1. 5 Informal codes of behavior 1. 3 Influences of corporate culture on the development of a company 1. 1. 1 Influences of a positive culture 1. 1. 2 Influences of a neutral culture 1. 1. 3 Influences of a negative culture . 4 Assessment of a company culture and phases to develop the desired one 1. 1. 1 Criteria to assess a company culture 1. 1. 2 Phases to develop the desired corporate culture CHAPTER 2: RESEARCH METHODOLOGY 2. 1 Study of structure in MCBS JSC 2. 2 Study of the company rules, regulations and procedures 2. 3 Study of the interactions between management and employees and between employees themselves CHAPTER 3: DISCUSION 3. 1 Strengths of MCBS JSC’s corporate culture 3. 2 Weaknesses of MCBS JSC’s corporate culture 3. 3 Recommendations to improve the company’s corporate culture How to cite Corporate Culture and Its Influence on the Development, Essay examples

Application of Integrated Marketing Communication †Free Samples

Question: Discuss About The Application Integrated Market Communication? Answer: Introduction Integrated Marketing communication is a process to plan the communications that provides business the energy to perform better which can happen from the campaigns and could reduce the costs of the marketing. Through the integration of different advertising tools such as social media, telemarketing and sales promotion, the need to provide clarity, impact of the maximum communication and also consistency can be provided (Bhupathi, 2016). It can create the awareness of the brand in a minimal cost among the customers. Organizations who are following the IMC is not only successful in promoting their brands among their targeted audience but also developed trust among the people whose interest towards only one brand is exhibited. IMC as well scores over the occidental ways of marketing strategies but also concentrates on winning the new customers and also focus on maintaining a healthy relationship based on longer term with the customers (Bhupathi, 2016; Luxton, Reid Mavondo, 2015). Woolsworths Group is one of the famous companies in Australia they focus on the building of culture and customer store led team. Woolsworth always focused on making its brand famous and they always work hard for 18 million customers and especially for those customers who have attraction for the same brand (Woolworths.com.au, 2017; (Bhupathi, 2016). Therefore, in this respect, Woolsworths Group has been chosen and its approach towards the Integrated Marketing Communication. A basic Model of Communication (Belch et al) There are two important participants who are being represented by the two elements in the process of communication and they are sender and receiver. The other two are important tools of communication such as messages and channels. Other four important functions of communication are encoding, decoding, response and feedback. The last element of the process of communication is the noise which indicates the non- essential factors which interferes with the work process against the effective means of communication (See Appendix). Source Encoding: The source or the sender is the organizations person who shares vital information with other group of people or other person. The source can be identified as a salesperson or a hired person, a celebrity who is paid to appear in the advertisement of the company or any kinds of organization such as corporations which are regarded as the non- personal entity. The process of communication takes place when the sources choose words, pictures, symbols in order to represent the message that it would get delivered to the receiver. The process of sharing communication with the other in the form of ideas, thoughts and information is known as encoding in a symbolic form (Belch Belch, 2016). Messages: The system of encoding gives birth to the message which includes the meaning or information that the source who wishes to convey to the receiver. The messages might be in the form of symbol, non-verbal oral or written. Messages must be in the format of transmittable which is suitable for the communication based channel. Channel: It is a method through which the communication takes place from either receiver or sender who are the sources. At the level of broadcast, communications channels are the types non- personal and personal. Communication based on personal channels which is carried on a direct basis interpersonal (face to face) contact with the groups or the targeted individuals (Belch Belch, 2016). The non- personal channels are those which carry out messages without any face to face contact between the receiver and sender. It is a channel through which the source transmits its messages to the targeted audiences or the group of people. For example, if a company chooses to sell its product, the famous companies or any start- up companies can do so through the attraction of media or through the means of mass communication. The non- personal channels include as well in the form of advertisements as well. Receiver/decoding: The receiver or the individual with whom the source or the sender shares thoughts and passes messages. In a general sense, the receivers are the consumers in the targeted market, who hears, reads or see the message of the marketers and decode the messages (Vernuccio, Ceccotti, 2015). Decoding is the process of transforming the messages of the sender into the thought process and ideas. This process is the significant way which influences the receivers experiences field and receiver, which describes the perceptions, attitudes, values that the receiver brings to the situation of communication. Decoding is an idea which gives birth to ideas in the head of the receiver about what the organization is intended to (Belch Belch, 2016). Noise: Throughout the process of communication, the messages are subjected to non-essential factors that interfere or distorts with the reception. The unsystematic distortion or the interference is referred to as noise (Belch Belch, 2016). Noise occurs because the experience of the sender and also the receiver do not overlap. The dearth of common ground may result to the improper ways of encoding the messages through the usage of symbol, words or signs which carry different meanings to the receiver. Feedback/Response: The set of reactions of the receiver after witnessing, reading or hearing the messages of the receiver is known by the term response. The response can be vary from the actions which are non- observable such as storing in memory the vital information such as dialing the toll number in order to buy the product by seeing the advertisements on television. The Marketers are interested in taking feedbacks, which is the part of the response of the receiver which is communicated back to senders. Feedback, can take a variety of forms and allow the sender to supervise how the message is intended and the process of being decoded and received by the receiver. Marking the targeted audience The process of marketing begins through the identification of the audience which would focus on the identified audience and also on the efforts of the promotion and the advertisements of the firms (Belch Belch, 2016). The Management which has been targeted may contain individuals who are having some specific needs and also for the people for whom the process of communication must be tailored. This also calls for the communication based on person to person and which can be achieved through the personal form of selling (Belch Belch, 2016). Process of Response: The most effective way to develop the process of communication incorporates the understanding the process of responses that the receiver may move through towards a particular behavior like the purchasing the products that they like and how the responses of the consumer are being influenced by the promotional efforts of the markets. Implementing Integrating Market Communication in Woolsworths Group IMC plays an important role in Woolsworth Group in the terms of Interactions which are based on Business to Business Channel communications of the Market Communications based on aim of the customer Communications which are internally directed (Woolworths.com.au, 2017; Luxton, Reid Mavondo, 2015). From the external point of view, which features the important points of IMC includes Advertising Internet/ social media (Schivinski, B., Dabrowski, 2016). Promotion of sales Developing relations with public Sponsorships and events (Woolworths.com.au, 2017). Internally carrying out communication takes place by the non executive director to the Director and CEO of group managing. The source over here is the Non- executive director who through the channel of communication send messages to the subordinates working under him such as Chief GM, Director Supermarkets, Logistics and Information officer and Chief Financial officer, further they carry out the communication to the other sources under their respective subordinates (Woolworths.com.au, 2017; Babin Zinkmund, 2015). Therefore, an integrated marketing communication is carried out inside the organization in this way. The administrative decision that is taken inside the organization gets effective outside the organization in terms of developing relations with the customers and making its brand famous (Woolworths.com.au, 2017; Andrews Shrimp, 2017). Integrating IMC The management of the Woolsworth Organization looks forward to place the preferences of the customers in the first position. They give advertisements to the public to make their brand famous in different websites, newspapers or any social media. The organization makes its channel of communication more non-personal in order to reach the customers more (Woolworths.com.au, 2017). The organization through the websites listens and gathers feedbacks about their stores and services. Later the receiver takes the feedbacks and work harder in order to improve the experiences of the customers. The Woolworth Organization practices Integrated Marketing Communication through the Customer Talkback and media press. Customer Talkative is regarded as a new form of initiation where the customers focus groups are being taken into account for a better understanding and take their feedbacks about their likings and disliking (Woolworths.com.au, 2017; Luxton, Reid Mavondo, 2015). The organization is regarded to be happy if the customers come with their feedbacks and more improvements are suggested to make the service special and helpful (Woolworths.com.au, 2017). The organization also believes in listening to the customers. The organization also focuses on the importance of their brand and sends the objectives through television, radio, cinema, digital and newspapers as mentioned before. They practice the communication with the customers through telemarketing, digital television for achieving direct responses and interaction. It maintains effective public relations which render help to organization and also in order to gain support from the public (Woolworths.com.au, 2017). For the sales promotion, the Woolsworth organization can also take these forms such as bonus, packs, coupons and competitions. Through the promotion of Events, to make their brand famous, it makes the customers engage and generate publicity (Kitchen Burgmann, 2015).In order to properly implement the IMC, it is important for the organization to being critical about the planning and process and monitor and control the communication system and leading to the proper IMC progr am. The Woolsworth Organization follows certain guidelines for the proper implementation of IMC They make sure that the IMC is practiced horizontally. They make communications design around the buying process of the customers. They develop a better information system of marketing. They follow clear objectives of communication and always link the values of brand with every communication. Ensuring visual standards are maintained in a common (Woolworths.com.au, 2017). Recommendations The marketing personnel, it is important for them to use the skills of people and also listen to their audience so that the requirements of the customers are addressed. The Woolworth Organization can create a short term marketing effects and promote the sales of the product (Kitchen Burgmann, 2015; Serrat, 2016). It is the responsibility of the company to check out the readiness of the buyers and the products that the customers are ready to buy through the process of the catchy messages such as knowledge, awareness, preferences and purchases. It is important for the company to set up budgets for the promotional purposes such as the budgeting has to be based on the requirements of the industry, advertisements for achieving the success based on promotion (Kitchen Burgmann, 2015; Carroll, 2015). The company must utilize its promotion mix with effective costing on direct marketing, online marketing or personal selling. The company needs to handle the different communication channels properly so that different communication channels do not show different messages. This might create a sense of misunderstanding among the customers which would make them not aware about the real scenario (Patti et al., 2017; Todorova, 2015). While doing online marketing, the company needs to be aware of ethical and legal issues which are subjecting the communications in market. The customer privacy is needed to be respected and the company must follow the social norms effectively for the growth of organization. Effective skills of communication would lead a faster, a better online marketing procedures (Fill Turnball, 2016; Argenti, 2015; Jackson Ahuja, 2016). Conclusion Therefore to conclude it could be said that Integrated Marketing Communication is a better way to understand the communication initiated by the organization to reach the customers to fulfill their targeted goals. The company Woolsworth uses an effective ways to promote communication to make their brands famous. Belch et al communication model provides an effective form of communication that an organization must practice to encourage the growth of the organization. It is a process which develops the strategy of marketing properly and makes an effective use out of it. Accordingly the organization follows the Integrated Marketing Communication by following this communication model and makes its brand name stronger with time. Finally the recommendations are given in terms of the model so that the company follows the IMC more properly in terms of their popularity. References Andrews, J. C., Shimp, T. A. (2017).Advertising, promotion, and other aspects of integrated marketing communications. Nelson Education. Argenti, P. A. (2015).Corporate communication. McGraw-Hill Higher Education. Babin, B. J., Zikmund, W. G. (2015).Exploring marketing research. Cengage Learning. Belch, G. E., Belch, M. A. (2016). Evaluating the effectiveness of elements of integrated marketing communications: A review of research.ISBN: 10 12-3869 Vol No 4 No 1 June 2016. Bhupathi, M. M. (2016). Integrated Marketing Communications. InStrategic Marketing Management in Asia: Case Studies and Lessons across Industries(pp. 425-445). Emerald Group Publishing Limited. Carroll, C. E. (Ed.). (2015).The handbook of communication and corporate reputation(Vol. 49). John Wiley Sons. Fill, C., Turnbull, S. L. (2016). Marketing communications: brands, experiences and participation. Jackson, G., Ahuja, V. (2016). Dawn of the digital age and the evolution of the marketing mix.Journal of Direct, Data and Digital Marketing Practice,17(3), 170-186. Kitchen, P. J., Burgmann, I. (2015). Integrated marketing communication: Making it work at a strategic level.Journal of Business Strategy,36(4), 34-39. Luxton, S., Reid, M., Mavondo, F. (2015). Integrated marketing communication capability and brand performance.Journal of Advertising,44(1), 37-46. Patti, C. H., Hartley, S. W., van Dessel, M. M., Baack, D. W. (2017). Improving integrated marketing communications practices: a comparison of objectives and results.Journal of Marketing communications,23(4), 351-370. Schivinski, B., Dabrowski, D. (2016). The effect of social media communication on consumer perceptions of brands.Journal of Marketing Communications,22(2), 189-214. Serrat, O. (2017). The future of social marketing. InKnowledge solutions(pp. 119-128). Springer Singapore. Todorova, G. (2015). Marketing Communication Mix.Trakia Journal of Sciences,13(1), 368-374. Vernuccio, M., Ceccotti, F. (2015). Strategic and organisational challenges in the integrated marketing communication paradigm shift: A holistic vision.European Management Journal,33(6), 438-449. Woolworths.com.au. (2017).{{metaController.metaData.title}}.Woolworths.com.au. Retrieved 23 August 2017, from https://www.woolworths.com.au/

Midterm Exam Business Valuation and Stock Valuation free essay sample

It is generally more expensive to form a proprietorship than a corporation because, with a proprietorship, extensive legal documents are required. (b) Corporations face fewer regulations than sole proprietorships. (c) One disadvantage of operating a business as a sole proprietorship is that the firm is subject to double taxation, at both the firm level and the owner level. (d) One advantage of forming a corporation is that equity investors are usually exposed to less liability than in a regular partnership. If a regular partnership goes bankrupt, each partner is exposed to liabilities only up to the amount of his or her investment in the business. (2) (TCO G) A security analyst obtained the following information from Prestopino Products’ financial statements: Retained earnings at the end of 2009 were $700,000, but retained earnings at the end of 2010 had declined to $320,000. †¢ The company does not pay dividends. †¢ The company’s depreciation expense is its only non-cash expense; it has no amortization charges. We will write a custom essay sample on Midterm Exam Business Valuation and Stock Valuation or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page †¢ The company has no non-cash revenues. The company’s net cash flow (NCF) for 2010 was $150,000. On the basis of this information, which of the following statements is CORRECT? (Points : 10) (a) Prestopino had negative net income in 2010. ( b ) Prestopino’s depreciation expense in 2010 was less than $150,000. (c) Prestopino had positive net income in 2010, but its income was less than its 2009 income. (d) Prestopino’s NCF in 2010 must be higher than its NCF in 2009. (e) Prestopino’s cash on the balance sheet at the end of 2010 must be lower than the cash it had on the balance sheet at the end of 2009. Which of the following is most likely to occur? (Points : 10) (a) The required rate of return for an average stock will increase by an amount equal to the increase in the market risk premium. (b) The required rate of return will decline for stocks whose betas are less than 1. 0. (c) The required rate of return on the market, rM, will not change as a result of these changes. (d) The required rate of return for each individual stock in the market will increase by an amount equal to the increase in the market risk premium. (e) The required rate of return on a riskless bond will decline.

PARAPHRASING Essays - Emotions, Happiness, Personal Life

PARAPHRASING Essays - Emotions, Happiness, Personal Life PARAPHRASING 1.Once a private act of perhaps the most intensely familial nature, eating together has become for many Americans a public act evolving others in a variety of relationships, few of which are matters of blood. James Helmer states that for many of the Americans, having food together, which has profounding familial nature, is a public performance of developing various new kinships, in which only a little is about blood relations. 2.McDonalds would win because it, better than any of its competitors, was able to catch up and exploit in its advertising a number of significant historical and contemporary development in American community and family life. The success of McDonalds advertisement is that, it succeeded in communicating and conveying about the remarkable historical and modern progress in American society and family life far better than any of its opponents. 3.Specifically, it demonstrates how familial images were employed as a means of persuasion that ultimately portrayed McDonalds as a potential source of love and human happiness as a place of being family. In advertisements, McDonalds presents various familial images, which depicts family as a place of love and affection of ultimate human happiness. 4.Family for some of these people no doubt meant these similar others, or perhaps the people with whom they worked or carried on some other activity. To few people in America family means, people who are alike them or may be people with they are engaging in some works or other activity. 5.What McDonalds would do was paint itself as many things that answered peoples basic and powerful need for someone to mend the fragments with caring. The action of McDonald is that it refines itself in many terms as to answer publics essential and strong requirements, which requires someone to restore broken pieces with care.

Dance teaching statement Personal Example | Topics and Well Written Essays - 500 words

Dance teaching - Personal Statement Example Neither does it purely encompass the sheer emotion obtained therein. It is more than just that. Dancing shapes one’s identity and personality and in essence, their destinies too. It leaves one completely transformed in a way that they will view certain important aspects of life in a different light that can only serve to advance them to better grounds and fortune. The process of learning dance itself is an unceasing activity that ensues throughout one’s life. It is therefore the work of the dance teacher to elicit this deep lifelong passion and determination in the students so that to the learner, dancing becomes a way of life and a vital necessity as taking breath. To achieve this desire in students, proper, endearing and encouraging communication with the students is inevitable and extremely relevant. As a dance teacher, I must strive first and foremost to evoke this deep connection in the students with the art of dance so that they can do their best without any coercion or feeling of duty. My students must enjoy dance and use the skills in every aspects of their livelihoods. It becomes ingrained into who they are and an effortless means of communication. I have learned and partaken of dancing for a period of two decades now. In addition to this, I have enrolled and completed courses in modern dancing in college for a period of four years. During this time, I have learnt a lot of modern dance techniques and can comfortably teach numerous arts such as the ballet with a modern touch to it. I want to teach solid and professional level ballet technique. My goal is to teach the importance of hard work in whatever aspects of life one is faced with through dance – be it in careers, studies, relationships, leadership, entrepreneurship, and so on. Dancing is life. The learning process and the practice of dancing are rich in information, solid

The Problem of Evil in a Divinely Created Reality Essay

The Problem of Evil in a Divinely Created Reality - Essay Example So, why do animals should feel pain and suffering? Why humans have to feel pain and suffering, even if their grand grandmother Eve was tempted by the Devil and now other people have to pay for her sins? Is God so cruel that he cannot forgive and forget? Why did he create a divine world, full of glory and bless if it is accessible only for those people, who did not make any sins? Evil is juxtaposition to deity. If there was no evil, people would not be able to compare it with the goodness or deity. Moreover, God created Heaven and Hell, life on the Earth and afterlife. There is an evident binary development of divine reality. Moreover, there are two types of evil: moral and physical. When committing a moral evil, a human being is committing it willfully (abortion, theft, murder etc); when committing physical evil, a human being commits it in the result of natural harm (such as famine, illness etc). The question is what for God created morally ill people or people, who are able to act in a morally evil way (Rachels and Rachels, 2008). Still, humans make their choice themselves. Moral evil is not instilled into people by God. God gives humans an opportunity to make a choice whether to commit evil or not; it is a test for morality and spiritual obedience. Therefore, moral evil is the result of our choice and it is another side of free will. Every person can make his own choice how to live, how to behave and if to commit evil or not. And of course, the further life of the person depends on the choice made. There are many other questions with respect to this issue, which deserve special attention and should be discussed. Now, let us consider whether suffering and pain are the same or different for people and animals (DeGrazia and Rowan, 1991). First of all, let us discard the notion that God is blamed for the existence of evil in the world. There was an intention creation of moral agents of God, who would supposedly create evil, but still these agents exist and for t heir evil deeds they suffer or experience pain. People may experience either physiological or psychological pain. These types of pain are different. The former kind of pain can be experienced by both humans and animals, while the second type of pain can be experienced only by humans. The concept of suffering has a more general meaning and it encompasses negative content of unpleasant emotions, anxiety, anger, fear, confusion etc (Derbyshire and Jones, 2011). Though suffering can be experienced without pain, it may trigger numerous factors of negative emotions or feelings. The scope of suffering is more impressive than the scope of negative experience, caused by pain. Therefore, God triggers negative emotions of humans in order to manipulate their actions and to direct the society in the right moral direction. At the same time he shows an option of deviance, which will lead to suffering and pain. There is no intention to punish humans or non-humans, there is an intention to support b inary relations on the Earth, which are determined between deity and evil. Life of humans and animals is a constant struggle for their existence and struggle against the surrounding dangers and threats. Thus, God does not intend to intimidate or destruct the humans and non-humans. He intends to make them use to realities of divinely created life in order they could survive in the real world. Works cited 1. DeGrazia, David and Rowan, Andrew. â€Å"Pain, suffering, and anxiety in animals and humans.†

Respiratory Syncytial Virus (RSV) Vaccine Formulation

Respiratory Syncytial Virus (RSV) Vaccine Formulation Abstract Respiratory syncytial virus (RSV) is the leading cause of upper and lower respiratory tract infections in both children and elderly people. In this study we evaluated the short- and long-term protective efficacy of the single intranasal immunization of RSV vaccine formulation consisting of the fusion (F) protein formulated with a polyIC, an innate defense regulator peptide and a polyphosphazene (ΆF/TriAdj). This vaccine induced significant systemic and local immunity, including RSV F-specific IgG1 and IgG2a, IgA and virus neutralizing antibodies, and IFN-ÃŽÂ ³-secreting T cells in mice. Furthermore, this vaccination promoted the induction of RSV F85-93-specific CD8+ effector T cells with no virus replication in the lungs. To evaluate the duration of immunity induced by single intranasal vaccination, mice were challenged with RSV after five months of immunization. Significantly higher levels of IgG1, IgG2a and virus neutralizing antibodies were detected in ΆF/Tr iAdj vaccinated animals. Moreover, this vaccine formulation induced high local IgA secreting memory B cell development and B cell IgA production and conferred complete protection against subsequent RSV challenge without priming for enhanced disease. In conclusion, a single nasal vaccination of RSV ΆF protein formulated with TriAdj enhanced robust, long- term protective immune responses against RSV infection. Introduction Respiratory syncytial virus (RSV) is a major cause of severe lower respiratory tract disease in infants, older and immunocompromised individuals worldwide {Falsey, 2005 #586}. Many RSV vaccine candidates, including live attenuated strains, vectored and viral protein subunit vaccines are underway. Currently, there is no licensed RSV vaccine or specific treatments other than prophylaxis with passive antibody therapy (Palivizumab) {Group, 1998 #599}. The first RSV vaccine clinical trial with formalin-inactivated RSV (FI-RSV) failed to induce neutralizing antibodies and caused enhanced respiratory disease after natural RSV infection {Kim, 1969 #589}. RSV is an enveloped virus, containing three transmembrane surface glycoproteins: the major attachment protein G, fusion protein F and small hydrophobic SH protein. The F protein is highly conserved and mediates fusion of the virus into host cells and subsequent formation of syncytia, making it a major target for subunit vaccine and antiviral drug development {Collins, 2008 #597}. However, mucosal vaccination with RSV F protein alone does not induce protective immunity {Vaux-Peretz, 1992 #648;Garlapati, 2012 #650} therefore, this protein needs to be combined with an adjuvant to induce strong, long-lasting protective immune responses. Previously, we developed a novel combination adjuvant platform (TriAdj) comprising of three components, namely a polyI:C, innate defense regulator peptide (IDRs) and polyphosphazene (PCEP). Synthetic double-stranded RNA, polyIC, known to recognize TLR3 and retinoic acid-inducible gene 1(RIG-I) leading to induction of proinflammatory cytokines, which in turn activates various immune cells {Sivori, 2004 #581`, Longhi`, 2009 `#573;Longhi, 2009 #573}. To stabilize and protect the polyIC from degradation, two other immunmodulators an innate defense regulator (IDR) peptide and a polyphosphazene were added. Innate defense regulator (IDR) peptides are derivatives of natural host defense peptides with microbicidal, chemotactic properties {Yeung, 2011 #608}. Poly[di(sodium carboxylatoethylphenoxy)]-phosphazene (PCEP) is a synthetic biodegradable water-soluble polymer with immunostimulatory properties. It forms non-covalent complexes with antigens and/or other adjuvants to increase their stabil ity and allow multimeric presentation {Andrianov, 2009 #618;Kovacs-Nolan, 2009 #621}. Previously, we developed a novel vaccine candidate against RSV (ΆF/TriAdj) consisting of a truncated version of the RSV fusion protein (ΆF) formulated with TriAdj. We have shown that double vaccination regime such as prime and boost intranasal immunization with this vaccine candidate establishes long-lasting humoral and cell-mediated protective immune responses without any evidence of immunopathology {Garg, 2014 #60;Garg, 2015 #63}{Garg, 2014 #60}{Garg, 2014 #60}{Garg, 2014 #60}{Garg, 2014 #60}{Garg, 2014 #60}. In the present study we demonstrated that after single IN delivery with ΆF/TriAdj, vaccine elicits mucosal and systemic immune responses and offers complete protection from RSV challenge in mice. Furthermore, ΆF/TriAdj induced effective local CD8+ T cells, which is one of the hallmarks for successful vaccination against many viral infections. To evaluate the duration of immunity induced by this vaccine candidate after single intranasal vaccination, we carried out long-term trials. Five month after the vaccination, RSV ΆF formulated with TriAdj induced robust mucosal and systemic immune responses and complete protection from RSV challenge in mice. Furthermore, ΆF/TriAdj elicited long-lived local IgA secreting memory B cell development and B cell IgA production, as well as memory T cells. It is well known that an RSV vaccine needs to induce a balanced Th1/Th2 adaptive immune response, which stimulates the generation of high-affinity neutralizing antibodies with effector cytotoxic CD8+ T cells {Graham, 2011 #642}. Results Single mucosal vaccination with ΆF/TriAdj induces strong systemic immune responses: Antibodies play a major role in protection against many viruses. To characterize the quality of the humoral immune responses induced by single vaccination of ΆF/TriAdj, IgG1 and IgG2a titers were measured in the serum. As shown in Fig. 1A B, the mice immunized with ΆF/TriAdj developed significantly higher IgG1 and IgG2a levels, than placebo groups both before and after challenge with RSV. These results indicate that formulation of ΆF protein with TriAdj promotes a balanced humoral immune response. Neutralizing antibody plays a protective role against viral infections. In order to evaluate the biological function of the ΆF-specific serum antibodies, Virus neutralization (VN) titers were determined. Mice immunized with ΆF/TriAdj developed significantly higher neutralizing antibody titers than the ones immunized with PBS both before and after challenge with RSV (Fig 1C). To investigate the phenotype of the cellular immune response, the ΆF-induced secretion of IFN-ÃŽÂ ³ and IL-5 by splenocytes was measured four days after challenge. Significantly higher numbers of IFN-ÃŽÂ ³ secreting cells were generated from mice vaccinated with ΆF/TriAdj formulation, while the number of IL-5 secreting cells was low in vaccinated mice (Fig 1D). This suggests the addition of RSVΆF protein to the TriAdj resulted in a stronger, Th1-biased immune response, which is in also agreement with the enhanced IgG2a production. Intranasal vaccination with single dose of ΆF protein formulated with TriAdj induces robust protective mucosal immune: Mucosal immune response play a crucial role in first line defense of host by blocking the spreading of RSV infection in the lungs. Thus, an effective RSV vaccine should induce local IgA antibodies in the respiratory tract. To evaluate the mucosal immune responses induced by ΆF/TriAdj, we measured ΆF-specific IgA antibody levels in the lungs. Significantly higher IgA production was observed in mice immunized with ΆF/TriAdj in comparison with mice immunized with PBS (Fig 2A). To determine whether single IN vaccination with the ΆF/Triadj formulations would affect viral clearance, all mice except those in one of the two Placebo groups were challenged IN with RSV. No infectious virus was recovered in lungs of mice immunized with ΆF/TriAdj, showing full protection from infection (Fig 2B). In order to investigate whether protection is correlated with higher cross-presentation and cell-mediated immune responses, we evaluated RSV F-specific CD8+ T cells infiltration with RSV F85-93 pentamers in the lungs after RSV challenge. The ΆF/TriAdj induced a significantly higher number of F85-93 specific CD8+ T cells influx in the lungs (Fig 6A), which suggests that vaccination with ΆF/TriAdj promotes a cytolytic CD8+ T cell response to RSV infection. Virus-specific effector CD8+ T-cell response has been shown to play a critical role in RSV clearance {Graham, 2011 #642}. Therefore, we analyzed the effecter function of the CD8+ T cells according to their expression of the IFN-ÃŽÂ ³ by à ¯Ã‚ ¬Ã¢â‚¬Å¡ow cytometry. After RSV challenge, the ΆF/TriAdj-immunized group showed higher frequencies of IFN-ÃŽÂ ³-expressing CD8+ T cells than PBS-immunized, RSV-challenged mice in the lungs (Fig 6). This suggests that ΆF/TriAdj promotes a stronger ef fector CD8+ T cell response to RSV. Single nasal vaccination with ΆF/TriAdj induces Long-term systemic immune responses: To assess the long term memory effect induced by single intranasal vaccination with ΆF/TriAdj, serum IgG and VN antibody levels were measured periodically over five months. The mice immunized with ΆF/TriAdj developed strong IgG and VN titers compare to placebo control group for approximately 25 weeks (Fig 3A B). We next examined whether IN vaccination promoted the induction of long term ΆF-specific serum IgG1 and IgG2a. The ΆF/TriAdj- vaccinated mice developed significantly higher ΆF-specific serum IgG1 and IgG2a titers than placebo groups both before and after challenge with RSV (Fig 4A B). VN titers were also determined to evaluate the functional quality of the ΆF-specific serum antibodies after RSV challenge. Mice immunized with ΆF/TriAdj developed significantly higher neutralizing antibody titers compared to PBS gro up (Fig 4C). To further define the phonotype of the immune response, the ΆF-induced secretion of IFN-ÃŽÂ ³ and IL-5 by splenocytes was measured four days after challenge with RSV. The ΆF/TriAdj formulation generated a significantly higher frequency of IFN-ÃŽÂ ³ secreting cells compared to the placebo. The numbers of IL-5 secreting cells were low in all vaccinated mice (Fig 4D), which suggests Th1 bias immune responses. Mucosal vaccination with Single dose of ΆF/TriAdj induces Long-term protective mucosal immune response: Local antibody play a major role in protection from respiratory viruses such as RSV in the lung. To access the long term local immune responses with ΆF/TriAdj vaccination, induction of ΆF-specific IgA in the lungs was examined by ELISA. Mice immunized with ΆF/TriAdj developed significantly higher IgA levels when compared to placebo and placebo virus challenged groups (Fig 5A). Furthermore, we checked the development of IgA-secreting memory B cells in lungs after RSV challenge. ΆF/TriAdj formulations generated a significantly higher frequency of RSV ΆF-specific IgA-secreting memory B cells in lungs in comparison to PBS (Fig 5B). In summary, single dose of ΆF/TriAdj via IN vaccination stimulated strong IgA-secreting memory B cell development and B cell IgA production. To determine viral clearance after single vaccination with ΆF/TriAdj, all mice except those in one of the two placebo groups were challenged IN with RSV on day 150. The mice were sacrificed after four days, and virus titers in the lungs were determined. No infectious virus particle was recovered from mice vaccinated with ΆF/TriAdj, showing that these mice still had sufficient immunity to be completely protected from challenge virus replication in the lungs (Fig 5C). Discussion RSV is one of the major global burdens of causing a broad spectrum of respiratory illnesses in children and older population worldwide. There is still no licensed RSV vaccine, in part due to the disastrous outcomes observed following vaccination of naÃÆ' ¯ve children with FI-RSV {Kim, 1969 #589}. Natural infection with RSV fails to protect against subsequent infection because it stimulate modest immunogenicity and short-lived immunological memory against virus. Similar to natural RSV infection, many promising RSV vaccine have failed to generate long-lasting, protective immune responses {Power, 2008 #769;Hall, 2001 #789}. For wide clinical application, RSV vaccine will need to be safe, easy to administer and must stimulate strong long-term protective immunity than natural RSV infection {Pulendran, 2011 #796}. Protein subunit vaccines have a high safety profile, but are generally poorly immunogenic and induce short-lived humoral and cellular immunity {Graham, 2011 #642}. So adjuvant s are added to subunit vaccines to stimulate faster, stronger, and long-lasting immune responses to vaccines. At the clinical level, multi dose vaccination is expensive and has potential safety problems. Whereas, single vaccination formats are cost effective, safe (no risk for vaccine contamination) and more convenient to health worker. In the present study, we demonstrated the short as well as long term protective efficacy including stable systemic IgG, local IgA production and neutralizing antibody titers of a RSV ΆF protein formulated with TriAdj in mice after single IN immunization. Furthermore, ΆF/TriAdj promoted an effector CD8+ T cells with compete protection against RSV infection without inducing vaccine-enhanced pathology in the lungs. A vaccine requiring only a single administration to be effective would be highly practical, as it would improve patient compliance Therefore, single intranasal immunization with RSV ΆF protein formulated with Tri Adj is a promising approach to induce a long-lasting protective RSV-specific immune response. Mucosal immunization via IN route is invasive and suitable for generation of both local and systemic immunity. RSV infect through respiratory tract, therefore mucosal antibodies specifically IgA play an important role in prevention and clearance of virus. Previously it has been shown that protection against RSV infection is more correlated to the levels of mucosal IgA rather than to systemic antibody in human {Watt, 1990 #687}. In mice, single RSV infection induces short-lived neutralizing antibodies with absence of antibody-secreting memory B cells {Schmidt, 2012 #869}. Interestingly, we have recently demonstrated that the RSV infected mice were showed significantly lower level of local B cell IgA production and IgA secreting memory B cell development that indicates an impaired local antibody response that allows RSV re-infection and explain the short-term protective immunity with natural infection with RSV{Garg, 2016 #1340}. In contrast, intranasal vaccination with single dose of R SV ΆF protein formulated with TriAdj promotes the induction of long-lasting local IgA secreting memory B cell development and B cell IgA production which is a major criterion for an effective RSV vaccine. Previously, we and others have reported that strong, effective and protective immune responses were induced only after a booster immunization with RSV F protein with adjuvant {Garg, 2014 #891}{Blanco, 2014 #1423}{Lambert, 2015 #1442}. A single mucosal vaccination of adenovirus-based RSV vaccine had also been proved effective in short term {Kim, 2010 #1443}. But this is the first report showing the long term protective efficacy of single mucosal vaccination of an adjuvanted RSV F protein subunit vaccine in mice. Clinical trial with FI-RSV failed to induce protection against RSV due to generation of poor neutralizing antibodies and TLR activation that led to vaccine-associated enhanced disease {Delgado, 2009 #513}. The inclusion of polyI:C as TLR ligand in vaccine formulation, is expected to overcome the limitations of inactivated RSV vaccines. The adjuvant effect such as long-lasting T cell immunity of polyI:C is likely caused by the direct interaction with PRRs such as TLR3, MDA5 and RIG-I, leading to production of pro-inflammatory cytokines, and chemokines. Interestingly, it has been shown that alveolar macrophages and lung DCs induced strong IgA and IgG antibodies by addition of TLR3 ligands, which were probably linked to secretion of BAFF/APRIL cytokines and activated B cells {Xu, 2008 #686}. However, we and others have been shown that IN administration of higher doses of polyIC caused marked production of inflammatory cytokines/chemokines accompanied by impaired lung function {Stowell, 2009 #682;Boukhvalova, 2010 #685;Aeffner, 2011 #750} suggesting excessive stimulation of local immune responses can result in detrimental effects. In contrast, ΆF/TriAdj has an advantage over polyIC alone as it has shown full protection form RSV infection without any exacerbation of chronic pulmonary disease. This demonstrates that a combination of polyIC, IDR and PCEP mediates optimal enhancement of the RSV-specific immune response without vaccine enhanced disease. We believe that IDRs is one of the major anti-inflammatory components in our vaccine formulation as it has been previously shown to enhance cell-mediated immune responses and to modulate excessive consequences of TLR signaling {Bowdish, 2005 #766;Hancock, 2006 #767}, whereas polyphosphazenes enhance antigen-specific humoral immunity through the formation of non-covalent complexes with protein {Andrianov, 2005 #667}, suggesting major roles from both of these compounds in this vaccine formulation. An important factor for successful vaccine against RSV requires the generation of effective CD8+ T cell in the lungs. Previously, it has been shown that mild to severe infection with RSV failed to induce virus-specific IFN-ÃŽÂ ³ recall responses in the infants {Lee, 2007 #806}. To define the stimulation of local effective CD8+ T cell responses induced by ΆF/TriAdj, RSV F specific pentamer staining demonstrated marked induction of IFN-ÃŽÂ ³ secreting RSV-specific CD8+ T cells in lungs. Similarly, exogenous IFN-ÃŽÂ ³ expression protects against RSV infection in the lungs of BALB/c mice {Kumar, 1999 #805}. This evidence suggests that complete protection against RSV was associated with the presence of IFN-ÃŽÂ ³+ RSV-specific CD8+ T cells. In summary, our data show that RSV ΆF protein formulated with TriAdj vaccine represents a safe, effective and promising RSV vaccine candidate. The major advantages associated with this mucosal RSV vaccine is to induction of long-term protective immunity with a single mucosal vaccination by stimulating long-lived RSV-specific neutralizing antibodies, memory B and CD8+ T cells, therefore warranting additional evaluation as a vaccine against RSV in clinical trials. MATERIALS AND METHODS Virus and vaccine formulation: The RSV (A2 strain) was propagated in Hep-2 cells (American Type Culture Collection, VR-1540). The RSV ΆF protein with his-tag was produced and purified as described previously {Garlapati, 2012 #650}. Briefly, HEK-293T cells were transfected with an episomal vector expressing the ΆF protein using Turbofect (Fermentas, R0534). The ΆF protein with his-tag was purified using TALON Superflow resin (Clontech,) according to the manufacturers instructions. The ΆF protein was formulated with 10  µg poly (I:C) (Invivogen, tlrl-picw), 20  µg IDR1002 (Genscript, 818360) and 10  µg PCEP (Idaho National Laboratory) in PBS (ΆF/TriAdj) as described previously. Animals, immunizations and challenge: Six to eight week old female BALB/c mice (Charles River Laboratories, Wilmington, MA, USA) were immunized once IN with ΆF/TriAdj. Two additional groups of mice received PBS IN (Placebo control). Three week (short-term trial) and twenty weeks (long term trial) post-vaccination, all except one of the Placebo groups were challenged IN with RSV strain A2 (5 x 105 PFU/50 µl). Mice were euthanized four days after RSV challenge for detection of immune response and virus in the lungs. Blood samples were drawn by cardiac puncture under anesthesia prior to challenge and at regular intervals afterwards. All procedures were approved by the University Animal Ethics Committee in accordance with the standards stipulated by the Canadian Council on Animal Care. Lung fragment cultures and ELISA: Lavaged lungs of euthanized mice were cut into small pieces and lung fragment cultures were prepared as described previously {Mapletoft, 2008 #591}. ELISA was performed as described previously {Garg, 2014 #942}. RSV ΆF-specific IgG1, IgG2a and IgA were detected using biotin-labeled goat anti-mouse IgG1, IgG2a (Southern Biotech, 1070-08, 1080-08) or IgA (Gibco, M3115) followed by streptavidin-alkaline phosphatase (AP) (Jackson ImmunoResearch Laboratories Inc.,016-050-084), and developed with p-nitrophenyl phosphate (Sigma-Aldrich, N3254) substrate. Enzyme-linked immunospot (ELISPOT) assays: ELISPOT assays were performed on splenocytes and LNs cells as previously described {Garlapati, 2012 #650}. Briefly, cells were stimulated with ΆF protein (2  µg/ml) and spots were developed using biotinylated IFN-ÃŽÂ ³- or IL-5- (BD Biosciences, 554410, 554397) or goat anti-mouse IgA specific antibody, AP-conjugated streptavidin and BCIP/NBT (Sigma-Aldrich, B5655) as the substrate. Analysis of CD8+ T cells from lungs by flow cytometry: Lung single-cell suspensions were generated as previously described {Garg, 2014 #942}. To analysis of RSV F85-93-specific CD8+ T cells, lung mononuclear cells were surface stained with H-2Kd-F85-93 MHC Class I pentamer (ProImmune, F149) together with labeled antibodies specific for CD8+ T cells (BD Pharmingen, 553031). For intracellular detection of IFN-ÃŽÂ ³ in CD8+ T cells, lung mononuclear cells were stimulated with 1 ÃŽÂ ¼M of F85-93 peptide (KYKNAVTEL) and cell surface staining was performed, followed by intracellular cytokine staining using Cytofix/Cytoperm (BD Pharmingen, 554715) and APC- conjugated anti-IFN-ÃŽÂ ³ antibody (BD Biosciences, 554413). After staining, cells were acquired by flow cytometry (BD Biosciences), and data were analyzed using Kaluza Software (Version 1.2). Cells were gated for live cells, singlets and lymphocytes and then analyzed for indicated markers. Virus titration and virus neutralization assay: Virus titrations were performed with individual lungs at day 4 post- challenge as described previously {Garg, 2014 #942}. Results are expressed as PFU/g of lung tissue. RSV-specific neutralization titers were determined by plaque reduction assays. Serum samples were mixed with 500 PFU/well of RSV strain A2 for 1 h at 37 °C. The sample-virus mixtures were transferred to HEp-2 cell monolayers and incubated for 4 days at 37  °C, and the cells were fixed and stained with 0.5% crystal violet. Statistical analysis: All data were analyzed using GraphPad PRISM version 6 for Windows (GraphPad Software, Inc., La Jolla, CA, USA). Differences among all groups were examined using Student t-tests, one-way ANOVA, followed by a Newman-Keuls post test. Differences were considered significant if P Acknowledgements The authors would like to thank Elisa C. Martinez, Wayne Connor, and Michael Theaker for technical assistance, as well as Sherry Tetland and Jan Erickson for the handling and care of the animals. This work was supported by the Krembil Foundation, and the Canadian Institutes of Health Research (CIHR). Published as VIDO manuscript number . Figure legends Fig. 1. RSV ΆF-specific systemic humoral immune responses in mice. (A) Serum IgG1 and IgG2a titers before challenge (B/C), (B) Serum IgG1 and IgG2a titers after challenge (A/C) with RSV, (C) Serum VN antibody titers determined before (B/C) and after (A/C) RSV challenge, and (D) Numbers of IFN-ÃŽÂ ³ and IL-5 secreting splenocytes determined in response to in vitro restimulation with ΆF protein. BALB/c mice were immunized once IN with ΆF formulated with TriAdj and challenged three weeks later with RSV. Control groups were immunized with PBS and challenged with RSV (Placebo) or mock-challenged (Placebo/mock). ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus neutralization titers are expressed as the highest dilution of serum that resulted in Fig. 2. Mucosal immune responses to RSV ΆF protein in mice after challenge with RSV. The IgA titer (A), Virus titer (B), Percentages of ΆF-specific CD8+ T cells (C), and ΆF-specific IFN-ÃŽÂ ³ secreting CD8+ T cells (D) were measured in the lung after RSV challenge. Mice were immunized and challenged as described in the legend for Fig. 1. ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus replication in the lungs is expressed as pfu per gram of lung tissue. Bars represent median values with interquartile ranges. *P Fig 3. Long-term systemic immune responses to RSV ΆF protein. IgG (A) and VN (B) titers were measured at different times after vaccination. BALB/c mice were immunized once IN with ΆF formulated with TriAdj and challenged with RSV on day 150. Control groups were immunized with PBS and challenged with RSV (Placebo) or mock-challenged (Placebo/mock). ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus neutralization titers are expressed as the highest dilution of serum that resulted in Fig 4. Systemic immune responses to RSV ΆF protein. (A) Serum IgG1 and IgG2a titers before challenge (B/C), (B) Serum IgG1 and IgG2a titers after challenge (A/C) with RSV, (C) Serum VN antibody titers determined after (A/C) RSV challenge, and (D) Numbers of IFN-ÃŽÂ ³ and IL-5 secreting splenocytes determined in response to in vitro restimulation with ΆF protein. Mice were immunized and challenged with RSV as described in the legend for Fig. 3. ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. Virus neutralization titers are expressed as the highest dilution of serum that resulted in Fig 5. Long term mucosal immune responses to RSV ΆF protein. IgA titers (A), numbers of IgA secreting LNs cells (B), Virus titers (C) were determined after (A/C) RSV challenge. Mice were immunized and challenged with RSV as described in the legend for Fig. 3. ELISA titers are expressed as the reciprocal of the highest dilution resulting in a value of two standard deviations above the negative control serum. IgA secreting cell numbers are expressed as the difference in the number of spots between ΆF-stimulated wells and medium-control wells. Virus replication in the lungs is expressed as pfu per gram of lung tissue. Bars represent median values with interquartile ranges. **P

Stefan’s Diaries: Origins Chapter 19

I galloped through the woods, kicking Mezzanotte to jump over logs, to dash through underbrush, anything to make sure I didn't lose sight of Katherine and Anna. How could I have trusted Katherine? How could I have thought I loved her? I should have killed her when I had the chance. If I didn't catch up to them, Anna's blood would be on my hands, too. Just as Rosalyn's was. We reached an uprooted tree and Mezzanotte reared up, sending me tumbling backward onto the forest floor. I felt a sharp stab as my temple cracked against a stone. The wind was knocked out of me, and I fought for breath, knowing it was only a matter of time before Katherine would kill Anna and then finish me off. I felt gentle, ice-cold hands lifting me up to a sitting position. â€Å"No †¦,† I gasped. The act of breathing hurt. My breeches were ripped, and I had a large gash on my knee. Blood flowed freely from my temple. Katherine knelt beside me, using the sleeve of her dress to stave off the bleeding. I noticed her licking her lips, then mashing them firmly together. â€Å"Y ou're hurt,† she said softly, continuing to apply pressure to my wound. I pushed myself away from her, but Katherine clasped my shoulder, holding me in place. â€Å"Don't worry. Remember. Y have my heart,† ou Katherine said, holding my gaze with hers. Wordlessly, I nodded. If death was to come, I hoped it would come quickly. Sure enough, Katherine bared her teeth, and I closed my eyes, waiting for the agonizing ecstasy of her teeth against my neck. But nothing came. Instead, I felt her cold skin near my mouth. â€Å"Drink,† Katherine commanded, and I saw a thin gash in her delicate white skin. Blood was trickling from the cut as though through a brook after a rainstorm. I was repulsed and tried to turn my head away, but Katherine held on to the back of my neck. â€Å"Trust me. It will help.† Slowly, fearfully, I allowed my lips to touch the liquid. Immediately I felt warmth run down my throat. I continued to drink until Katherine pulled her arm away. â€Å"That's enough,† she murmured, holding her palm over the wound. â€Å"Now, how do you feel?† She sat back on her heels and surveyed me. How did I feel? I touched my leg, my temple. Everything felt smooth. Healed. â€Å"Y did that,† I said incredulously. ou â€Å"I did.† Katherine stood up and brushed her hands together. I noticed her wound, too, was now completely healed. â€Å"Now tell me why I had to heal you. What are you doing in the forest? Y know ou it's not safe,† she said, concern belying her chiding tone. â€Å"Y †¦. Anna,† I murmured, feeling sluggish ou and sleepy, as one might feel after a long, wine- infused dinner. I blinked at my surroundings. Mezzanotte was hitched to a tree, and Anna was sitting on a branch, hugging her knees to her chest and watching us. Instead of terror, Anna's face was full of confusion as she looked from me, to Katherine, then back to me. â€Å"Stefan, Anna is one of my friends,† Katherine said simply. â€Å"Does Stefan †¦ know?† Anna asked curiously, whispering as if I wasn't standing three feet from her. â€Å"We can trust him,† Katherine said, nodding definitively. I cleared my throat, and both girls looked at me. â€Å"What are you doing?† I asked finally. â€Å"Meeting,† Katherine said, gesturing to the clearing. â€Å"Stefan Salvatore,† a throaty voice said. I whirled around and saw a third figure emerge from the shadows. Almost without thinking, I held up the vervain from my breast pocket, which looked as useless as a daisy clutched in my hand. â€Å"Stefan Salvatore,† I heard again. I glanced wildly between Anna and Katherine, but their facial expressions were impossible to read. An owl hooted, and I pressed my fist into my mouth to keep from screaming. â€Å"It's okay, Mama. He knows,† Anna called to the shadows. Mama. So that meant Pearl was also a vampire. But how could she be? She was the apothecary, the one who was supposed to heal the sick, not tear out human throats with her teeth. Then again, Katherine had healed me, and she hadn't torn out my throat. Pearl emerged from between the trees, her gaze tightening on me. â€Å"How do we know he's safe?† she asked suspiciously, in a voice that was much more ominous than the polite tone she used at her apothecary. â€Å"He is,† Katherine said, smiling sweetly as she gently touched my arm. I shivered and clutched the vervain, Cordelia's words echoing in my head. This herb could stop the devil. But what if we'd all gotten it wrong, and vampires like Katherine weren't devils but angels? What then? â€Å"Drop the vervain,† Katherine said. I looked into her large, cat-like eyes and dropped the plant to the forest floor. Immediately, Katherine used the tip of her boot to cover it with pine needles and leaves. â€Å"Stefan, you look as though you've seen a ghost,† Katherine laughed, turning toward me. But her laughter wasn't mean. Instead, it sounded melodic and musical and slightly sad. I collapsed onto a gnarled tree root. I noticed my leg was shaking and held my hands firmly against my knee, which was now completely smooth, as if the fall had never happened. Katherine took the motion as an invitation for her to perch on my knee. She sat and looked down on me, running her hands through my hair. â€Å"Now, Katherine, he doesn't look like he's seen a ghost. He's seen vampires. Three of them.† I glanced up at Pearl as if I were an obedient schoolboy and she were my schoolmarm. She sat down on a nearby rock slab, and Anna perched next to her, suddenly looking much younger than her fourteen years. But, of course, if Anna was a vampire, then that meant she wasn't fourteen at all. My brain spun, and I felt a momentary wave of dizziness. Katherine patted the back of my neck, and I began to breathe easier. â€Å"Okay, Stefan,† Pearl said as she rested her chin on her steepled fingers and gazed at me. â€Å"First of all, I need you to remember that Anna and I are your neighbors, and your friends. Can you remember that?† I was transfixed by her gaze. Pearl then smiled a curious half smile. â€Å"Good,† she exhaled. I nodded dumbly, too overwhelmed to think, let alone speak. â€Å"We were living in South Carolina right after the war,† Pearl began. â€Å"After the war?† I asked, before I could stop myself. Anna giggled, and Pearl cracked a tiny sliver of a smile. â€Å"The War of Independence,† Pearl explained briefly. â€Å"We were lucky during the war. All safe, all sound, all a family.† Her voice caught in her throat, and she closed her eyes for a moment before continuing. â€Å"My husband ran a small apothecary when a wave of consumption hit town. Everyone was affected–my husband, my two sons, my baby daughter. Within a week, they were dead.† I didn't know what to say. Could I say I was sorry for something that had happened so long ago? â€Å"And then Anna began coughing. And I knew I couldn't lose her, too. My heart would break, but it was more than that,† Pearl said, shaking her head as if caught in her own world. â€Å"I knew my soul and my spirit would break. And then I met Katherine.† I glanced toward Katherine. She looked so young, so innocent. I glanced away before she could look at me. â€Å"Katherine was different,† Pearl said. â€Å"She arrived in town mysteriously, without relatives, but she immediately became part of society.† I nodded, wondering who, then, was killed in the Atlanta fire that brought Katherine to Mystic Falls. But I didn't ask, waiting for Pearl to continue her story. She cleared her throat. â€Å"Still, there was something about her that was unusual. All the ladies and I talked about it. She was beautiful, of course, but there was something else. Something otherworldly. Some called her an angel. But then she never got sick, not during the cold seasons, and not when the consumption began in town. There were certain herbs she wouldn't touch in the apothecary. Charleston was a small town then. People talked.† Pearl reached for her daughter's hand. â€Å"Anna would have died,† Pearl continued. â€Å"That's what the doctor said. I was desperate for a cure, wracked with grief and feeling so helpless. Here I was, a woman surrounded by medicine, unable to help my daughter live.† Pearl shook her head in disgust. â€Å"So what happened?† I asked. â€Å"I asked Katherine one day if she knew of anything that could be done. And as soon as I asked, I knew she did. There was something in her eyes that changed. But she still took a few minutes of silence before she responded and then–â€Å" â€Å"Pearl brought Anna to my chambers one night,† Katherine interjected. â€Å"She saved me,† Anna said in a soft voice. â€Å"She saved me,† Anna said in a soft voice. â€Å"Mother too.† â€Å"And that's how we ended up here. We couldn't stay in Charleston forever, never growing old,† Pearl explained. â€Å"Of course, soon we'll have to move again. That's the way it goes. We're gypsies, navigating between Richmond and Atlanta and all the towns in between. And now we have another war to deal with. Seeing so much history really proves to us that some things never do change,† Pearl said, smiling ruefully. â€Å"But there are worse ways to pass the time.† â€Å"I like it here,† Anna admitted. â€Å"That's why I'm scared we'll be sent away.† She said that last part as a whisper, and something about her tone made me achingly sad. I thought of the meeting I'd attended that afternoon. If Father had his way, they wouldn't be sent away, they'd be killed. â€Å"The attacks?† I asked finally. It had been the one question that had been nagging at me ever since Katherine's confession. Because if she didn't do it, then who †¦ ? Pearl shook her head. â€Å"Remember, we're your neighbors and friends. It wasn't us. We never would behave like that.† â€Å"Never,† Anna parroted, shaking her head fearfully, as though she were being accused. â€Å"But some of our tribe have,† Pearl said darkly. Katherine's eyes hardened. â€Å"But it's not just we or the other vampires who are causing trouble. Of course, that's who everyone blames, but no one seems to remember that there's a war going on with untold bloodshed. All people care about are vampires.† Hearing Damon's words in Katherine's mouth was like a bucket of cold water in my face, a reminder that I wasn't the only person in Katherine's universe. â€Å"Who are the other vampires?† I asked gruffly. â€Å"It's our community, and we will take care of it,† Pearl said firmly. She stood up, then walked across the clearing, her feet crunching on the ground until she stood above me. â€Å"Stefan, I've told you the story and now here are the facts: We need blood to live. But we don't need it from humans,† Pearl said, as if she were explaining to one of her customers how an herb worked. â€Å"We can get it from animals. But, like humans, some of us don't have self-control, and some of us attack people. It's really not that much different from a rogue soldier, is it?† I suddenly had an image of one of the soldiers we'd just played poker with. Were any of them vampires, too? â€Å"And remember, Stefan, we only know some. There could be more. We're not as uncommon as you may think,† Katherine said. â€Å"And now, because of these vampires we don't even know, we're all being hunted,† Pearl said, tears filling her eyes. â€Å"That's why we're meeting here tonight. We need to discuss what to do and come up with a plan. Just this afternoon, Honoria Fells brought a vervain concoction to the apothecary. How that woman even knows about vervain, I have no idea. Suddenly, I feel like I'm an animal about to be trapped. People have glanced at our necks, and I know they're wondering about our necklaces, piecing together the fact that all three of us always wear them†¦.† Pearl trailed off as she raised her hands to the sky, as if in an exasperated prayer. Quickly, I glanced at each of the women and realized that Anna and Pearl were wearing ornate cameos like the one Katherine wore. â€Å"The necklace?† I asked, clutching my own throat as if I, too, had a mysterious blue gem there. â€Å"Lapis lazuli. It allows us to walk in daylight. Those of our kind cannot, usually. But these gems protect us. They've allowed us to live normally and, perhaps, even allowed us to stay more in touch with our human side than we would have otherwise,† Pearl said thoughtfully. â€Å"Y don't know ou what it's like, Stefan.† Pearl's matter-of-fact voice dissolved into sobs. â€Å"It's good to know that we have friends we can trust.† I took out my handkerchief from my breast pocket and handed it to her, unsure what else I could do. She dabbed her eyes and shook her head. â€Å"I'm sorry. I'm so sorry that you have to know about this, Stefan. I knew from the last time that war changes things, but I never thought †¦ it's too soon to have to move again.† â€Å"I'll protect you,† I heard myself saying, in a voice that didn't quite sound like mine. â€Å"But †¦ but †¦ how?† Pearl asked. Far off in the distance, a branch broke, and all four of us jumped. Pearl glanced around. â€Å"How?† she said again, finally, when all was still. â€Å"My father's leading a charge in a few weeks.† I felt a tiny pinprick of betrayal as I said it. â€Å"Giuseppe Salvatore.† Pearl gasped in disbelief. â€Å"But how did he know?† I shook my head. â€Å"It's Father and Jonathan Gilbert and Honoria Fells and Mayor Lockwood and Sheriff Forbes. They seem to know about vampires from books. Father has an old volume in his study, and together they came up with the idea to lead a siege.† â€Å"Then he'll do it. Giuseppe Salvatore is not a man to have his opinions easily swayed,† Pearl stated. â€Å"No, ma'am.† I realized how funny it was to call a vampire ma'am. But who was I to say what was normal and what wasn't? Once again, my mind drifted to my brother and his words, his casual laughter when it came to Katherine's true nature. Maybe it wasn't that Katherine was evil, or uncommon at all. Maybe the only thing that was uncommon was the fact that Father was fixated on eradicating the vampires. â€Å"Stefan, I promise that nothing I've said to you was a lie,† Pearl said. â€Å"And I know that we will do everything in our power to ensure that no more animals or humans are killed as long as we're here. But you simply must do what you can. For us. Because Anna and I have come too far and gone through too much to simply be killed by our neighbors.† â€Å"Y won't be,† I said, with more conviction ou than I ever had in my life. â€Å"I'm not sure what I'll do yet, but I will protect you. I promise.† I was making the promise to the three of them, but was looking only at Katherine. She nodded, a tiny spark igniting in her eyes. â€Å"Good,† Pearl said, reaching out her hand to help a sleepy-eyed Anna to her feet. â€Å"Now, we've been here in the forest too long. The less we're seen together, the better. And, Stefan, we trust you,† she said, just the tiniest hint of a warning in her otherwise rich voice. â€Å"Of course,† I said, grabbing Katherine's hand as Anna and Pearl walked out of the clearing. I wasn't worried about them. Because they worked at the apothecary, they could get away with walking in the middle of the night; they could easily tell anyone who saw them that they were searching for herbs and mushrooms. searching for herbs and mushrooms. But I was scared for Katherine. Her hands felt so small, and her eyes looked so frightened. She was depending on me, a thought that filled me with equal amounts of pride and dread. â€Å"Oh, Stefan,† Katherine said as she flung her arms around my neck. â€Å"I know everything will be fine as long as we're together.† She grabbed my hand and pulled me onto the forest floor. And then, lying with Katherine amid the pine needles and the damp earth and the smell of her skin, I wasn't frightened anymore.