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How to create an outline for your research paper

What is the first step of research paper writing? Preparing an outline for the paper. If you have a research outline ready before

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How to structure a research paper

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business plan sketch 285 days before open. You have gotten a business idea. You must transform it into how to structure a research, a first sketch. In short, this quick sketch enables you to research made easy, determine if your business idea is realizable by you. Structure A Research Paper. Have you the financial capacity to realize it? Is the idea profitable? You must answer to research made, these two crucial questions before engaging you in more complex studies. External readings: 0. Do it yourself: About 21 hours. Our objectives are to show you: -How to draw up a first description of how to structure a research, your product and organization according to your business idea. -How to estimate your financial capacity for realizing it and the profitability. By the end, you will know if it's worth to go ahead and to begin more extensive studies.

In case of deserted island definition, positive answer, you will implement the first steps of structure paper, your business plan. 1-FIRST SKETCH: DEFINITION. The sketch describes: - your service or your good in connection with your expected market and your competitive advantage. - The organization and the skills you need. -The costs and the financial capacity required. Down earth advice: A business idea can be great. It does not mean that you are able to realize it.

May be you have gotten a great idea about the steel industry or the space exploration but it requires huge capital requirement and literature critical coursework, you will never be able to structure, collect the planning research paper money! The fist sketch just takes place after the business idea. It corresponds to how to a research paper, a logical progression . Look at the John progression: -Firstly , he has a vision : I want to create a large open air tourism company -According to this vision he has a business idea: I want to essay introduction, set up a travel agency near the Perito Moreno glacier to show visitors round the glacier. We can expect that this business idea is good: It should be good if john has a competitive advantage : For example, john has practiced mountain climbing for years and he can lead the tourists in some place which are inaccessible's for other agencies. How To Structure A Research Paper. Secondly, there is a market : For example there are visitors all along the introduction year and due to his competitive advantage john can target another segment of this market: experienced alpinists.

Clearly, John must now describe his idea into how to structure, a sketch: What is the service really provided? Is it a week trek in the mountains or a daily trip to the glacier? What is the organization he needs? Does he need a large office and a lot of vehicles and power vs authority, drivers? What are the starting costs? Does he get the financial capacity for structure paper covering these starting costs? Is the idea profitable? According to some quick calculations how much profit could he get in essay, such a business?

2-DESCRIPTION OF THE GOOD OR SERVICE. Of course, in this description you will constantly target the customer benefit . Paper. You must constantly put yourself in the customers' shoes and know what he really wants, and the price he is prepared to pay. The marketing study must identify this problem. Essays Papers Quickstudy Academic. For the time being, you should know roughly what type of customer you will target. Secondly, along this description you will constantly keep in mind your competitive advantage. It means that all the benefits you provide to structure a research paper, the customer must be focused on this competitive advantage. Clearly, you must enhance this advantage. You must therefore describe: - The nature of the service or product. -The value it brings to the customer. -The life of the product. -The product guarantee, the after sales service and so on. In our John example, a visit to a glacier can be a weekly trek or just a daily trip.

Let 'suppose that it's a daily trip. you must then give details of what the customer wants in the form shown in the table below: - The customer wants to set off from his hotel at 7 a.m. Meet him at the hotel reception area. - The customer wants to have a cup of coffee during the journey. Power Vs Authority. Serve him a cup of coffee. - The customer wants to structure paper, talk. Make conversation with him. - The customer wants to stop for & term papers quickstudy academic a break. How To A Research Paper. Include this in you timing. - The customer wants to literature anthology coursework, drink an aperitif. . Provide drinks and biscuits. Paper. You give details about the quickstudy academic service. You time each sequence.

You draw up an overall schedule for the day. You will have to structure a research, modify this description according to the further studies but at this stage, you must have a detailed hypothesis. - Once you have chosen your offer and identify the tasks it implies you have to note down the means required next to research paper, each task: Transportation to the glacier, lunch, duration of the visit and so on. Go back to the previous table about the glacier visit. - Meet the customer. A driver and a guide. A flask of coffee. The guide must speak several languages as well as being educated. A Research. - Serving an aperitif. The driver serving the your research aperitif must be trained. -Once you have described your core offer, you have to think about its environment: For example, you must implement a light organization for accounting , invoices, bank relations and so on.

Any light organization may imply a secretary with fax, telephone and computer. What is more, it needs also a location. In short, you list all the tasks and means and structure, you break each problem into its components parts. -Then you have to your research paper, think about a research paper some possible hurdles: The John's project can harm environment, disturb wild animals or bring troubles to local population. John must study the product or service's conformity with regulations ( the service must respect national park regulations). Is the guide job regulated? does he need an official diploma. What are the insurances to subscribe and so on. Down earth advice. Beware of persuasive essay, these regulations that many people ignore.

Take care of them right now. A great business idea can only fail on how to structure a research some specifics point that the & term quickstudy promoter had forgotten to check up. The french use to say that the devil is into the details. We suppose that you have identified the tasks and all the means you need such as equipment, human resources and so on. Then you will estimate the how to structure costs of these different means in distinguishing the start up period and then the running period. 411-Identifying the papers costs. According to your description of the tasks and how to structure a research, means, You list all your costs per unit, per month, and then on several months in distinguishing the start up period and then the running period. In the travel agency example, let's suppose that we have the following costs in essays papers quickstudy academic, $: -Rent a location in how to structure paper, town: 600 per month. -A secretary for administrative work: 1000 per month. -A driver: 200 per month. -A guide: 600 per month. Power Vs Authority Essay. -Consumables: 1150 per structure, month. -A computer: 1000 (Purchase price on catalog) -A vehicle with little equipment: 20200 (idem) -Promotion: 10000 during the starting period for launching the literature product.

412-Starting and running costs. In this example, we suppose that you will not get cash receipts during the six first months. Structure A Research. It's the essays & term papers quickstudy starting period. All the costs during this period should be covered by the money you bring for how to structure a research starting a biz. On the contrary, by planning your paper the end of this period you will get receipts from the customers. It is the running period. The costs occurring during the running period are calculated on 12 months . You can now list the human resource, together with their Cost, using the previous list.

The costs can be calculated approximately from the labor market situation and social regulations. item---- monthly cost---- starting cost---- running cost. Consumables disappear as soon as they have been used, such as , renting of the premises, energy, petrol, drinks, coffee. Item-------- monthly cost--- Starting costs--- running costs. Carry out the same calculation for Equipments, i.e. goods you can use for several years. Categories------- Starting period------- Running period. This evaluation shows that you must have $52500 in cash for covering investments and costs during the starting period . If we add the miscellaneous (10% of the structure a research paper former total), we reach $60000.

Once you have done that, I pretend that in essays quickstudy, fact you have done nothing! you have just set up a shopping list. Any low skilled person should be able to do the structure paper same job. Of course, these costs are certainly too high compared to the money you can invest . A2 English Literature Critical. Disheartened, you will give up. This hopeless scenario affects 9 out of 10 promoters. As you are following a training, you will therefore think differently: You have to show your creativity. -Firstly examine any item and ask yourself the following question: What service is provided for the customer? How much is the structure a research customer prepared to pay for essays & term quickstudy this service? What benefit do I gain from how to a research this cost? By asking questions in this way, you will notice that certain costs are unnecessary. -Secondly examine the price of each item.

There are the catalog price for power vs authority the equipments and the market labor salaries for the human resource . Now, apply to these costs the following golden rules: -Equipments: Whenever I can, I rent. How To A Research Paper. If I can't rent, I buy second hand on the international market . You would be surprised to find just how many machines are for sale for peanuts! -Staff: I only hire people on open-ended contracts. I try to use temp agencies. I make use of teleworking : It exists today on the market self employed secretaries or drivers. -Globally: Whatever the mean (Equipment or staff) I do not buy, I do not recruit: I just assemble.

I only pay a material or a person for the period I need them. A2 English Critical. If you apply these golden tips, be sure, you will halve the costs of how to, your operations. Instead of 60000, you will finally reach 30000! We reach now a critical moment. Persuasive Essay. You must now check that you have the financial capacity to realize the project and secondly that it has a reasonable profitability. If the project does not answer both to these two questions, it should be worthless to go on with detailed studies going nowhere. I will give you two quick tests for answering these crucial questions: 51-Financial feasibility: The starting equity/starting cost ratio . The starting equity/starting cost ratio is an easy tip that we have invented for simplifying your job.

Let us imagine that you have $10 000 to invest from how to structure paper your savings. According to your calculations, t he Starting project cost is: Below $10000 : No problem. You are in the good lane. Between 10 and 15 000: It is certainly possible to delay some payments. Papers. No real problem. Between 15 and 20 000 : You need Associates. You will have to find extra money from friends and relatives. If you have already some hopes in how to paper, this domain go ahead. Between 20 and 30 000 : Go back to paper, your business idea and make some changes: For example, Instead of taking visitors round the how to a research Perito Moreno glacier 50km from the town, organize hiking to show them the lake and its wildlife.

You can take notice that your vision remains unchanged. Above 30 000$: Go straight back to the creativity module. Your business idea cannot be realized and there is no point continuing. Keep your vision because you should never spoil it but find a new business idea. It means that whatever your savings S, the evaluation method is very straightforward: Starting costs less than 150% of S: Go ahead Starting costs between 150% to 200% of S: Look for associates. Starting costs between 200% to your, 300% of S: Bring change to your business idea. Starting cost above 300% of S: Find another business idea.

You can think that this method is conservative because it does not take in account venture capital and bank loans. In fact, a bank only lends 1$ per 1$ of paper, your personal equity. You will never be able to borrow 3$ for 1$ of persuasive essay introduction thesis, equity! Down earth advice: Do not believe in the fairy tales. Statistics show that for a start-up like you, the chances of obtaining a loan are about 0.5%! What is structure paper, more, the formalities are very lengthy.

Do not allow 1 year to create your business, it is more likely to take you 2 or 3 years! Do not think that the fact to begin little condemn you to made, remain small for ever. Constantly keep in your mind the structure paper following facts: -$1 that makes 20% a week turns into planning research paper, $13 000 after a year: $10000 would therefore earn you $ 130 million in a year! -Small equity x Creativity = Big money. Everything depends on your creativity. Creativity is in fact the real big equity that you need! 52-Profitability feasibility: The running costs/selling price test.

For testing the how to structure paper profitability, people use frequently the break even analysis . You will find it in further module. For the deserted present moment you have not yet a detailed market study or a detailed analysis of structure a research, all your producing costs. You need a quick test to paper easy, just see if you are in the good way or in the wrong one. How To A Research. For this purpose, we currently use the Running cost/Selling price test or alternatively the Running costs/Customers test. Persuasive Introduction. -Firstly, just take the total of your yearly running costs . In our example 42600. - Secondly observe the average price on the market for how to structure paper the type of service you intend to sale . In our example, let's suppose that a day trip to the glacier is a2 english literature critical anthology coursework, currently priced $60 per person. - Then just apply the following formula: 42600 /60= 710 . This means that you must have 710 customers to recover your total running costs without making any profit . Structure Paper. With 700 customers you go in red. With 720 you begin to make profit. Now let's suppose that the your research paper total number of visitors in paper, the entire area amounts 600 per deserted definition, year! It means that you will never reach 710 visitors and that your project has not any chance to how to a research, get a profitability. -If you are alone on the market because your product is new, you have not any average market sale price. -In this case, determine an expected number of visitors . You can have an idea about it according to your production capacity.

For example, with one vehicle, you can only manage two visitors per day. If the essays papers academic season lasts 6 months, it means that you can manage: ------------ 2 visitors* 20 days a month* 6 months = 240. -Then, apply the formula: 42600/ 240= 177. Structure A Research Paper. This means that you must price $177 per introduction thesis, person the day trip to the glacier for recovering your total running costs without making any profit . Obviously, this selling price is not reasonable. It's not worth to spent your time in more complex studies. You have to reduce your costs and may be to change your business idea. You have a business idea.

You must establish a simple first sketch. This sketch describes the good or the service you will offer. It gives a short description of the organization required and lists the how to structure a research tasks and the means. It enables you to papers quickstudy academic, estimate the cost of all the means. The grid cost analysis distinguishes starting costs and running costs . Finally, you can check the global feasibility of your project: The Starting equity/ Starting cost ratio enables you to check your financial capacity to structure, do the a2 english critical anthology project and the Running costs/Selling price ratio provides you with a quick test about its profitability. You have to establish: 1-A description of the how to a research paper good or service you are going to papers academic, offer.

2-A description of all the tasks you have to perform for offering this good or service: 3- A description of all the how to structure a research paper means you need for starting and then running the biz: Consultants, human resources, equipment etc. 4-A short description of your organization. 5-A grid analysis of the costs : It's the difficult part of the job and you need to collect some information's around you: -About human resources. Make a list of self employed staff you might need: Lawyers, accounting firms . Ask about their fees. Pay a visit to temp agencies. Ask about power vs authority essay their tariffs. About consultants, go to: . This site provides list of how to structure a research, consultant in every domain and the volunteers of Score who are mainly retired executives can provide you with free advices trough e-mails. Look at the large temp agencies and ask for their prices: -About material and equipments.

Ask your local chamber of commerce for second hand equipment. Go to and & term papers quickstudy academic, click on Business and a research, industrials. You will find here all type of materials and equipments to buy for a dime! 6- Then do the two feasibility tests . If their results are positive, begin to planning research paper, write down your sketch. You will use it for how to a research paper writing your executive summary which currently is the opening of anthology, your business plan. How To. 7-Open your business plan. Your business plan is your road map. It is made up of chapters and spreadsheets. Along your next studies, you will enter their results in paper, the right chapters. What is more you will use the spreadsheets for your calculations.

The best way is to use some stuff already existing on the web. I have selected for how to a research you the following sites: go to . Your. Click on business planning freeware and download a free business plan frame. For the present moment, just include your business idea in how to structure, a first chapter. A2 English Critical Anthology Coursework. Add right now a second chapter titled Business description and how to paper, report inside the description of the tasks, the short organizational sketch and your grid analysis. Theses papers should be removed or modified ,step by step, with the further courses. This job could take 20 hours or more according to your project. A complex project could take three times more: The grid cost analysis needs frequently a long time. Of course computers facilitate but the anthology coursework analysis for reducing the costs currently take a bit of time! You could send for examination and advice your business idea and your sketch.

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The sector as a whole currently employs 1,126,000 people (approximately 4% of the UK workforce) in around 230,000 businesses. In addition, there are an structure, estimated 500,000 volunteers working in the sector on a regular basis. Approximately 42% of the workforce is self-employed. The animal care industry offers opportunities to work in kennels or catteries, through to animal charities, zoos and wildlife parks. The work in this industry generally covers the essay thesis, health, welfare, handling supervision and husbandry of animals. Animal care encompasses: pet shops; animal boarding establishments, animal breeding; dog training (including animals in entertainment and how to a research, education, as well as working dogs); pet styling and vs authority essay, grooming; animal welfare charities; animal care services (for example animal hydrotherapy, dog walkers/sitters); zoos and wildlife parks (including wildlife rehabilitation); animal visitor attractions; and animal related public services (for example dog/animal wardens). There are approximately 78,000 people working in the industry, in around 12,650 businesses. Almost three quarters of the animal care industry accounts for pet shops and pet supplies, boarding establishments, and dog clipping and grooming businesses. 86% of businesses employ 5 or less staff. 87% of the workforce is a research employed full-time. A2 English Critical Coursework! Volunteers are a significant part of the how to structure, workforce within the animal care industry.

Jobs in the industry include: animal trainer (audio visual), assistance dog trainer, head zoo keeper, dog trainer – racing greyhounds, inspector, dog trainer – uniformed forces, animal care assistant, animal clinic assistant, dog groomer, manager of zoos or wildlife establishments, safari park ranger, dog warden. East Midlands – There are an estimated 6,750 employees in the regional workforce, in around 1,100 businesses. Thesis! East of England – There are an estimated 9,300 employees in the regional workforce, in around 1,450 businesses. London – There are an estimated 8,750 employees in the regional workforce, in around 1,000 businesses. How To A Research Paper! North East – There are an estimated 2,100 employees in the regional workforce, in around 450 businesses. North West – There are an estimated 6,100 employees in the regional workforce, in around 1,300 businesses. South East – There are an estimated 11,200 employees in the regional workforce, in around 1,850 businesses. South West – There are an estimated 9,400 employees in the regional workforce, in around 1,450 businesses. Power Vs Authority! West Midlands – There are an estimated 7,300 employees in the regional workforce, in around 1,050 businesses. Yorkshire and the Humber – There are an estimated 5,700 employees in how to structure a research, the regional workforce, in around 1,050 businesses. Northern Ireland – There are an definition, estimated 1,100 employees in structure paper, the regional workforce, in persuasive thesis, around 250 businesses.

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essay on structure paper hvtn 505 The path forward to an efficacious HIV-1 vaccine will be expedited via the analysis of a2 english immune correlates from past and future HIV-1 vaccine efficacy trials. How To A Research Paper! Approaches that aim either to improve upon current findings from a partially efficacious vaccine strategy or approaches toward the essay generation of broadly neutralizing antibodies by rational immunogen design strategies are likely to a research paper, be fast-tracked through careful probing of the intersections of power vs authority essay immune correlate analyses within and among human vaccine clinical trials. When carefully interrogated for correlates of structure a research paper risk (immune responses and host/virus genetics), each of the anthology coursework efficacy trials (failed or partially efficacious) can provide new information to advance the HIV-1 vaccine development field. In some cases, clues about potentially protective immune responses can be discerned, and of equal importance, potentially detrimental or distracting immune responses can be identified. Immunogen design strategies may benefit from these analyses, such that that immunogens can be paired down to structure, essential components. Research! Moreover, insights from these studies on how host and how to structure a research virus genetics play a role in vaccine-induced immunity will enable the advancement of immunogen designs toward globally efficacious vaccines across different populations. The intersection of correlate analyses or studies involving immune responses (innate, cellular, humoral), host genetics (e.g., FcR expression, HLA) and HIV-1 virus sieve analyses (virus sequence analysis of all infections (placebo and vaccine groups) to determine if there were specific genetic attributes of infecting viruses that were significantly different between the two groups) are areas of the highest information content for made, follow-up studies (Figure 1).

In this review, the correlate analyses for how to structure a research, HIV-1 vaccine efficacy trials are described, with an emphasis on those studies that examine the intersection of different components and persuasive introduction thesis analyses. Due to structure paper, the uniqueness of each efficacy trial and outcome, each study can provide information regarding the minimum bar to overcome to achieve HIV-1 vaccine efficacy. Even when there is no overall vaccine efficacy, the heterogeneity of immune responses among vaccines has enabled follow-up studies to identify associations of research made immune responses with HIV-1 infection risk in subsets of vaccines. Furthermore, an integral part of understanding immune correlates of protection is to also define patterns of immune responses that associate with non-protective vaccines. One of the how to structure major goals for the HIV-1 vaccine field is to island, define correlates of a research protection from HIV-1 infection, such that the field has biomarkers that will clearly predict vaccine outcome in persuasive essay introduction thesis HIV-1 vaccine efficacy trials.

There have been six HIV-1 vaccine efficacy studies to date [1,2,3,4,5,6] (Table 1), each testing either a different vaccine strategy, or different populations with different risk factors and geographic locations. HIV-1 vaccine efficacy studies have been analyzed for correlates of infection risk; however, a correlate of protection for an HIV-1 vaccine has not yet been identified. Both the how to structure a research non-efficacious vaccines [7,8,9,10,11] and essay introduction the one partially efficacious HIV-1 vaccine [12,13,14,15] trial have yielded several correlates of a research paper infection risk; although more weight is island definition given to how to structure paper, those findings from trials where there was overall vaccine efficacy. In total, three of power vs authority essay these studies (VAX004, Step, and RV144) found significant correlations with HIV-1 infection risk/incidence and two of these studies (Step and RV144) identified potential sites of structure immune pressure on the virus (virus sieve). The terminology employed for vs authority essay, describing correlates and a research surrogates for protection from HIV-1 infection have been the focus of power vs authority essay ongoing discussion and clarification [16,17,18,19]. It is generally agreed upon that for any one of the correlates of infection risk identified in how to structure a research RV144 to be considered a correlate of protection that reliably predicts protection from essays papers academic HIV-1 infection, it must be tested and proven in paper another clinical trial either with the same setting or in a different vaccine setting [18]. Ultimately, it is the power vs authority latter confirmation of the how to a research paper correlate to essay, HIV-1 vaccine efficacy in a different geographic region, different virus strains, different risk groups, etc . that is the structure a research most sought after for the development of a global HIV-1 vaccine. (Efficacy futility determined at first interim analysis after full enrollment ) Vaccine-induced immune responses have been studied to identify immune correlations with infection risk and evidence of virus sieve that can inform the design and evaluation of the next phase of vaccine efficacy trials (shaded boxes indicate reported correlations). Immunizations in research made two efficacy studies (Phambili/HVTN 503, HVTN 505) were stopped; however, follow-up of participants continues. How To Structure Paper! a Vaccine Efficacy (VE) Outcome is noted as “No Efficacy” if there was no overall statistically significant vaccine efficacy; b MSM: Men who have sex with men; c Participants, meeting enrollment criteria, were enrolled from the general population; d TG: Transgender.

3. Cellular Immunity and HIV-1 Vaccine Efficacy Trials. HIV-1 specific CD8 + T cells are associated with control of HIV-1 replication as demonstrated in acute HIV-1 infection [26,27,28,29,30] (reviewed in McMichael et al. [31]) and in studies of anthology those rare individuals who can naturally control HIV-1 infection long term [32,33,34,35,36,37]. Moreover, studies in NHP have demonstrated, as proof of concept, that vaccine induced CD8 + T cells can be protective [38,39,40,41]. In two HIV-1 efficacy trials (Step and Phambili) aimed at inducing CD8 + T cell responses (in the absence of HIV-1 envelope specific antibodies), there was no overall association with vaccine efficacy. However, these studies indicated that there were vaccine-elicited CD8 + T cells that could impact virus replication; although not sufficient enough to structure paper, provide overall decreased risk of infection in the trial (either due to island definition, low magnitude/breadth/function of T cell responses and/or pre-existing vector immunity etc. ). HVTN 505, also designed to target the induction of how to structure paper CD8 + T cell responses (as well as HIV-1 envelope specific antibodies), was discontinued and unblinded due to lack of paper efficacy.

Studies of the magnitude and how to structure a research quality of the literature critical CD8 + T cell response are ongoing. Since there was evidence of increased infections in how to paper two of the Ad5 vector based vaccine studies, Step and Phambili [42] but not in HVTN 505 [6], there is a greater emphasis on understanding immune responses to research paper made easy, vaccine vectors and how to paper impact on literature critical anthology subsequent immunity. Additional studies demonstrated that there may be some cross-reactivity in the T cell responses among the different adenovirus serotypes that will have to be tested carefully going forward as vectors are chosen for clinical trials [43]. An alternate interpretation is how to structure that perhaps many other types of HIV-1 vaccines induce some level of CD4 + T cell activation; however, the key is to have sufficient overall HIV-1 specific immunity to tip the balance toward protection. Despite these setbacks on vs authority understanding how to induce protective CD8 + T cell induced immune responses in human HIV-1 clinical trials, a new paradigm for structure a research paper, T cell immunity has emerged from deserted island NHP vaccine studies.

Picker and colleagues report on how to structure a research a CMV based vaccine regimen that protects NHPs from infection and induces a novel subset of CD8 + T cells that can broadly recognize HIV-1 epitopes through class II restriction [38]. The series of papers by Picker and colleagues provide new hypotheses to test for inducing effective cellular immunity, both in terms of a2 english literature critical both new immunogen vector design and new subsets of CD8 + T cells to target with vaccine approaches. HVTN 502 (Step) and how to structure a research HVTN 503 (Phambili); Host Genetics and & term academic Virus Sieve Analyses. The Step study [3,44] as well as the Phambili study [4] were test of how to structure concept studies (double-blind, randomized, placebo-controlled) for induction of HIV-1 specific T cells responses. These trials tested the concept of a replication defective adenovirus serotype 5 (Ad5) vector with clade B HIV-1 genes ( gag , pol nef ) (MRKAd5) to decrease virus load in those who became infected. This vaccine regimen generated antigen specific T cell responses, but overall these responses did not result in vaccine efficacy. An increased rate of essay HIV-1 infection was observed in vaccines compared to placebos that was associated with being uncircumcised and having pre-existing Ad5 antibodies (reviewed in Gray et al. [45]). This increased risk was confirmed in follow-up studies that also reported a waning of the increased infection risk over time [46]. In order to understand the lack of efficacy (and enhancement of infection in a research the vaccine group), follow-up studies were undertaken to understand the a2 english anthology immunity that was generated by vaccination. One of the goals of these follow-up studies was to structure a research paper, examine potentially protective responses that may be subdominant in the population but may impact either acquisition or disease progression in some of the vaccines. Research Made! Notably, detailed analysis of the “breakthrough” viruses, or viruses that did establish infection despite vaccine-induced immunity, demonstrated that there was selection at specific sites indicating potential vaccine induced T cell specific immune pressure [23].

Notably, the a research paper antigen specific CD8 + T cells responses induced by Ad5 vaccination were to essay, HIV-1 epitope hotspots that were different than those generated in natural infection and were also directed to highly variable regions that likely can tolerate escape mutations from paper vaccine induced immune responses [47]. Host genetics also played a role in immunity, since those vaccines with HLA alleles (B*27, B*57, B*58:01), known to be associated with HIV-1 control, had lower viral load [48] and their CD8 + T cells exhibited greater killing in power essay in vitro assays [49]. Recent analysis of how to structure a research those infected within a year of their last vaccination, identified that total T cell breadth and total magnitude of the immune response after the immunization series and prior to infection significantly correlated with lower mean viral load [50]. A sieve analysis of the Step trial found that there was significant sequence divergence of the persuasive introduction infecting viruses compared to the vaccine [23]. Structure! However, despite an your paper observed anamnestic T cell response (from vaccine-induced immunity) after infection, the specific T cell responses measured as part of the how to a research paper evaluation of the trial did not correspond with the sieve findings [51]. Moreover, none of the HIV-1 antigen specific T cell responses that were tested were associated with risk of infection in the vaccine recipients [11]. Surprisingly, the non-HIV specific ELISpot magnitude was a significant direct CoR for HIV-1 infection in a2 english literature critical anthology coursework the vaccine group. How To A Research! Thus, sieve analyses along with immune correlate analyses can be informative in generating new hypotheses about baseline or vaccine-induced responses correlating with infection risk.

4. Humoral Immunity and HIV-1 Vaccine Efficacy Trials. Four of the HIV-1 vaccine efficacy trials induce HIV-1 Env-specific antibody responses by three diverse strategies: recombinant HIV-1 gp120 immunogen alone (VAX003 and VAX004); vector prime with gp120 boost (RV144); and DNA prime, vector boost (HVTN 505). Although VAX003 and VAX004 efficacy trials each tested gp120 protein only as an immunogen strategy to induce humoral responses, these trials were distinct in that they were conducted in different risk populations (injection drug users (IDU) vs . men who have sex with men (MSM)) and with different clades of gp120 protein immunogens (B/E vs . B/B). Similarly, both RV144 and HVTN 505 were prime boost strategies, but each tested a different vector (ALVAC vs . rAd5) and in paper different risk populations (community-based risk in Thailand vs . MSM in the U.S.). It is unknown whether the same mechanisms of protection against paper HIV-1 acquisition will be similar or quite distinct among the different risk populations/transmission routes. Thus, there are four major differences across these HIV-1 vaccine efficacy trials that need to be considered when comparing studies: (1) differences in the HIV-1 gp120 sequence content; (2) differences in the approach (prime/boost and research easy vector vs . protein); (3) diverse infection risk populations tested by how to structure a research each vaccine; and (4) different geographic locations and clades of HIV-1. 4.1. VAX003 and island definition VAX004: Immune Response and Host Genetics. The VAX003 clinical trial was conducted in high-risk injection drug users, and the vaccine contained bivalent gp120 (Clades B and how to structure a research paper Clade E: AIDSVAX B/E) [2].

As part of the primary analysis of critical coursework vaccine efficacy for VAX003, pre-specified antibody variables (V2 and structure V3 binding antibodies to critical, a vaccine strain envelope (A244), gp120 binding antibodies (A244 and MN), CD4 blocking antibodies and neutralizing antibodies to HIV-1 MN) were found not to be correlates of risk. The VAX004 clinical trial enrolled high-risk men and women (predominantly men who have sex with men) and a research paper the vaccine contained two clade B gp120s (AIDSVAX B/B) [1]. Follow-up studies of deserted definition vaccine-induced humoral responses in VAX004 reported that higher neutralizing antibody (nAbs) responses to an easy to neutralize virus [7], CD4 blocking antibodies [7], and antibody-dependent cellular virus inhibition (ADCVI) [8] correlated with lower HIV-1 infection risk among those in the vaccine group. Neutralizing antibody responses against more difficult to how to structure a research paper, neutralize viruses ( i.e ., circulating viruses that are the target of vaccine strategies) were also induced. However, one interpretation is that this level and breadth of response may be below the bar needed for efficacy (and, thus, providing some information on where the bar sits) [52].

Comparative studies among vaccine efficacy trials can also inform as to what might constitute a potentially protective immune response. Neutralizing antibody responses were higher in VAX003 compared to RV144 [53], indicating that higher antibody responses were not by themselves better nor predictive of a positive outcome. Paper! Insights from analyses of how to host genetics suggest that although there was no overall increased infection in the VAX004 vaccine trial, the Fc? receptor IIIa genotype (VV genotype, encoded by an allele with a valine (V) a position 158) was associated with an increased rate of island HIV-1 infection in a research low risk vaccines, but not in high risk vaccines [9]. RV144 is the power vs authority essay only HIV-1 vaccine efficacy trial in which the vaccine showed decreased transmission versus the placebo group, with an estimated vaccine efficacy of a research paper 31% [5]. Essays Quickstudy Academic! An international collaborating team of how to structure a research scientists completed a case control study reporting that two humoral immune measurements correlated with the risk of HIV-1 infection [12]. IgG antibodies to the V1/V2 region of HIV-1 gp120 correlated with a decreased risk of infection [12,13,14,15], and a plasma HIV-1 envelope-specific IgA score correlated with increased risk of infection in the vaccine arm (decreased vaccine efficacy) [12,54]. Six primary variables were tested in the correlate analyses: plasma Env IgA score, Env IgG avidity, HIV-1 neutralizing antibodies (nAbs), antibody-dependent cellular cytotoxicity (ADCC), V1/V2 Env IgG and Env-specific CD4 + T-cells.

Overall, humoral responses were the predominant immune response in deserted this trial, with evidence of vaccine-elicited V2 targeted CD4 + T-cell responses [12,55]. One key insight from the evaluation of the structure paper RV144 vaccine trial was that the vaccine immunogen was unique in its antigenicity ( i.e . , exposure of specific epitopes) compared to other envelope proteins [25,56]. Studies of the RV144 protein immunogen demonstrated that certain epitopes were better exposed, as a result of an 11 amino acid N -terminal deletion of the AE.A244 gp120 envelope, thus leading to the induction of dominant V1V2 antibody epitope specificities that were well exposed on the vaccine immunogen [56]. & Term Quickstudy! In particular, the A244 gp120 used in RV144 was antigenic for structure a research, both linear V2 epitopes bound by paper strain-specific V2 antibodies, as well as for conformational V1V2-glycan epitopes bound by V1V2 broad neutralizing antibodies (BnAbs) [25,56]. Thus, 8,000 individuals have been immunized with an antigen expressing a BnAb epitope, yet only non-glycan-dependent nAbs were induced [12,25,57,58]. Thus, in how to a research addition to persuasive essay, defining correlates of protection in RV144, a second critical reason to study vaccine responses in RV144 is to understand why BnAbs were not induced by an antigenic immunogen. How To! These findings indicate that careful evaluation of envelope antigenicity ( i.e ., determining what epitopes are exposed for immune recognition) can provide insights into research paper, the types of antibodies that may be elicited by vaccination. 4.2.2.

RV144: Intersection of Antibody Epitope Specificity, Antibody Function and Antibody Form. Further analyses of the six primary variables in the RV144 correlate analyses using interaction models was one way of examining the role of multiple immune measurements in how to assessing correlates of HIV-1 infection risk. The secondary analysis as part of the RV144 immune correlate work [12] indicated (through a statistical interaction model) that in the presence of essays & term quickstudy academic low vaccine-elicited IgA responses, either ADCC or neutralizing antibody responses correlated with a decreased risk of how to structure a research infection. The ADCC responses induced by RV144 were predominantly to the C1 conformational region of gp120 [59,60]; among other epitope specificities ( i.e ., V2, V3) [25]. Thus, one hypothesis from the RV144 correlate study was that C1 region Env-specific IgA could block C1-specific ADCC by binding to the same epitopes on deserted definition infected cells, but without the structure paper functional engagement of NK cells that is power essay needed to mediate ADCC. Indeed, RV144-induced Env IgA was shown to block C1 region-specific IgG-mediated antibody-dependent cellular cytotoxicity by natural killer cells [54]. These studies are an example of how the intersection of a research paper multiple approaches can provide insights into vaccine efficacy.

Specifically, this work highlights the need to examine vaccine-induced epitope-specific antibody responses that engage different cellular Fc receptors. 4.2.3. RV144 Secondary Analyses: Supporting the Primary Correlates and Further Hypotheses. Together, the two immune correlates of HIV-1 infection risk in RV144, along with data from the secondary analyses and follow-up studies, indicate that evaluating antibodies that target circulating strains in vs authority essay the population being tested as well as cross-clade epitopes may play a role in understanding vaccine efficacy. Circulating Env IgA responses to the C1 region in gp120 of a research paper a circulating strain in paper Thailand (CRF01_AE) was the strongest statistical correlate of HIV-1 infection risk in a research paper RV144.

Moreover, IgG responses to the V2 region of CRF01_AE Env significantly correlated with a decreased risk of vs authority essay infection among the different clades of how to paper V2 responses tested in the peptide microarray assay [12,15]. Further analyses indicate that the breadth, or cross-reactivity to multiple clades, of the V1/V2-specific response induced by RV144 [13,14,25,57,58] may have contributed to paper, the unique nature of the vaccine-induced immune response in how to structure a research paper RV144. Further studies in your paper other vaccine trials will be able to test these specific hypotheses to see if vaccine-induced immunity that can broadly target circulating strains will be critical for how to, vaccine-mediated protection from HIV-1 infection. The RV144 case control study was designed to paper easy, evaluate potential correlates of infection risk. Notably, multiple analyses support the original primary RV144 immune correlates and follow-up studies involving immune responses and/or host genetics relate to the primary and how to secondary correlates analysis (Table 2). These studies indicate that other humoral responses and host genetics may modulate vaccine-induced immunity and impact vaccine efficacy. Virus sieve analyses have complemented humoral immune data demonstrating that different approaches together can provide stronger clues to vs authority essay, potential mechanisms for vaccine efficacy. How To Paper! Although there were only six primary immune measurements tested in power vs authority essay the case control analysis, other measurements as part of the RV144 case control study [12] and as follow-up analyses [13,14,15,54] were significantly associated with HIV-1 infection risk (statistical results with p 0.05) (Table 3). Structure Paper! Notably, the plasma Env IgA correlate of critical risk was specific to certain HIV-1 Env IgA responses, as not all Env IgA correlated with decreased vaccine efficacy [54] as measured by a binding antibody multiplex assay [61]. 4.2.4.

RV144 Studies: Virus Sieve Analysis and B-Cell Repertoire. For HIV-1 vaccines, immune responses that impact only a proportion of the transmitted viruses can lead to virus sieving, in that specific viruses impervious to vaccine-elicited immunity can establish infection. Paper! These virus sieve analyses in concert with probing vaccine-induced immune response can lead to hypotheses of a2 english anthology coursework specific immune responses that may prevent the acquisition of some viruses. Moreover, analyses of structure a research paper viruses escaping from vaccine-induced immunity can provide a roadmap for critical targets on infectious virions that need to paper easy, be targeted by an HIV-1 vaccine to substantially increase vaccine efficacy. Structure Paper! Ongoing analyses of the changing complexity of circulating viruses may be critical for deserted island definition, understanding the virus target in specific populations. Additional independent studies examining breakthrough viruses in RV144 vaccines (sieve analyses) and how to structure paper the generation of monoclonal antibodies from the B-cells of vaccine recipients have pointed toward a specific site within the V2 region of the HIV-1 envelope. V2 mAbs derived from island definition RV144 vaccines bound to an epitope centered on K169 of the V2 region [25], and a sieve analysis [24] focused on this region found K169 to be a critical site of immune pressure. Thus, if the infecting strain of HIV-1 matched the RV144 vaccine AE.V2 region at K169, the vaccine efficacy was 48%.

Thus, we have proposed that one way to enhance the efficacy of follow-up trials to how to structure paper, RV144 would be to include additional Envs or V2 peptides with V2 sequences not covered in the original RV144 envelope sequences. Studies of such combinations are currently underway in non-human primates [62]. The functional properties of research paper these V2-specific antibodies include ADCC, neutralization and low level virus capture [25,63]; however, it is unclear if these findings related to V1/V2 IgG may be mechanistic or non-mechanistic correlates [19]. Thus, the how to a research paper combination of the initial analysis of the V2-directed humoral responses induced by RV144, work identifying that V1V2 (K169) IgG3 correlated with a decreased risk of infection [14], the mAb work that identified the importance of a lysine in position 169 of the V2 loop and & term papers academic virology work that resulted in the demonstration of K169 as a site of immune pressure, together, reinforced the hypothesis that the structure a research induction of V2-specific antibodies is important to test as a correlate of protection in further HIV-1 vaccine efficacy trials. (1) V2-specific mAbs were isolated and characterized, which involve position 169 in the V2 loop for binding [25]. (2) V1/V2 Env IgG responses are robustly correlated with a decreased risk of infection (multiple V1/V2 antigens and two different assays and laboratories) [13,57]. (3) linear V2 IgG by peptide microarray correlates with a decreased risk of infection [15]. (4) linear V2 IgG by the binding antibody multiplex assay correlates with a decreased risk of infection in RV144 [64]. (5) IgG3 to V1/V2 CaseA2_169K correlates with a decreased risk of infection [14]. The six primary immune measurements evaluated in the RV144 correlate analysis were chosen in part based on each one representing distinct immunological space. Power Vs Authority! Two of these measurements (V1/V2 IgG and Env IgA) significantly correlated with the risk of HIV-1 infection as reported in how to paper Haynes et al [12].

Additionally, the reported results of interaction models indicate potential relationships among the six measurements. Below are the three interactions that were statistically significant. IgG Env avidity. Neutralizing Antibody (NAb) Score. A series of a2 english literature coursework immune measurements, in addition to the preselected primary six variables, were evaluated for correlations with the how to a research risk of HIV-1 infection. Of all the secondary immune variables, eight of these had a p -value of 0.05. Three of these are related to the direct correlation with plasma Env IgA; two are related to research paper made, the indirect correlation with Env IgG responses, and three are related to cellular responses ( i.e ., peripheral blood mononuclear cell (PBMC) derived cytokines). A series of immune measurements, in a research paper addition to the preselected primary six variables, were evaluated for correlations with the risk of your research paper HIV-1 infection. Of all the secondary immune variables, eight of these had a p -value of 0.05. Three of these are related to a research paper, the direct correlation with plasma Env IgA; two are related to the indirect correlation with Env IgG responses, and three are related to cellular responses ( i.e ., PBMC derived cytokines).

4.2.5. & Term Papers Quickstudy! Multiple Antibody Specificities and Function: Vaccine Efficacy. Due to structure, the natural redundancy of the immune system [18], HIV-1 vaccines may induce multiple potentially protective immune responses. HIV-1 vaccines can induce a broad repertoire of HIV-1 epitope specificities and antibody isotypes/subclasses with a variety of different antiviral functions. Essays & Term Quickstudy Academic! Even in RV144, where V1/V2 Env IgG responses were identified as an immune correlate of risk of how to a research HIV-1, there was broad heterogeneity among vaccines in their immune responses with a diverse repertoire of antibody forms, specificities and functions [63].

Some of these diverse immune measurements were not tested in the immune correlates analysis, due to research made easy, inherent assay limitations, sample limitations and the requirement of minimizing test variables in how to structure a research order to maintain statistical power. The interaction models from RV144, as well as several RV144 follow-up studies that examine several antibody specificities and/or function [54] support the hypothesis that multiple antibody specificities with different functions may act in concert. Evaluating multiple antibody specificities and planning your research antiviral functions, in addition to identifying single correlates of how to structure infection risk, will be central to identifying mechanistic correlates of protection for HIV-1 toward the goal of identifying an efficacious vaccine strategy. 5. Time-Dependent Immune Measurements and Vaccine Efficacy. One important insight from vaccine efficacy studies is the impact of time on vaccine-induced immunity. The time point at paper made easy, which vaccine efficacy is determined in a trial can dramatically influence the resulting level of a research paper vaccine efficacy. RV144 had a 60.5% vaccine efficacy through 12 months after initial vaccination that declined over time [70]. The reported vaccine efficacy was pre-specified to be reported at 42 months post follow-up and was 31.2% at that time . This higher level of vaccine efficacy at an earlier time point is hypothesized to be due to a higher level or quality of antibody response that decreased over island definition, time For example, Env IgG3 responses are induced in RV144, but decline more rapidly than the overall Env IgG response [14]. Current vaccine strategies aim to improve the durability of the anti-envelope antibody response through different adjuvant/immunogen combinations. Thus, the half-life of a research paper Env-specific antibody responses will likely be a critical component for evaluating further HIV-1 vaccine candidates.

Vaccine-induced cell-mediated immunity also depends on time post-vaccination. Cellular immune responses, as well as humoral responses, can wane post-vaccination. A recent study [50] reported that CD8 + T-cell immunity that correlated with decreased virus load post-breakthrough infection was potentially related to time since vaccination. HIV-1 vaccines that can induce an orchestrated response comprised of all arms of the immune response (innate, cellular and humoral) may prove to thesis, be the most efficacious across different populations, genders and routes of transmission. Thus, it is imperative to advance forward with strategies for a research paper, both effective T- and power essay B-cell responses that likely will be induced through specific engagement of the innate immune system. New technologies and approaches have allowed the field to how to structure a research, cast a broader net in evaluating vaccine strategies.

The advent of virus sequencing at vs authority, the single genome level has allowed for a detailed analysis of the specificities of how to structure immune responses that can impact virus replication in both human and & term non-human primates. These virus-specific approaches for that are not sensitive to vaccine-induced immunity compared to those viruses that are sensitive; (2) potential insights into important vulnerable sites (and their structure) on the virion that vaccines should target; and (3) information on whether the described vaccine-induced humoral and structure a research paper cellular immune responses have epitope specificities that match those identified as sieve sites in the virus. New approaches towards understanding systems biology contributes to a better understanding of the interplay between the your research vaccine-induced host immune response and virus. Taken together, these technological advances have yielded a robust amount of how to structure a research data that has an unprecedented need for innovation in the way the data are analyzed. Analysis of deserted island host genetics, viral genetics and immune responses [71,72] have already provided numerous insights into how to structure paper, both innate and adaptive responses of virus control and disease progression in HIV-1 infected individuals. Follow-up studies provide evidence of an interplay of vaccine-induced immunity against the background of host genetics. Understanding the literature anthology coursework role of host genetics and the functional attributes of vaccine-induced immunity can enable a better understanding of structure a research paper which immune correlates will be broadly applicable in a globally efficacious vaccine.

Similarly, studies are ongoing to understand if immune responses to essay introduction thesis, one vaccine can be predicted by immunity to structure a research paper, another vaccine; thus, also allowing deeper insights into intrinsic population effects that may not be distinct to the vaccine strategy being tested. Currently licensed vaccines for other pathogens are based on empirical studies, and many do not have defined mechanistic correlates of protection. Persuasive! The protective efficacy of some of these vaccines are associated with antibody responses, with the exception of rotavirus and BCG ((Bacillus Calmette-Guerin) tuberculosis vaccine) [18,73]. Notably, these antibody responses are of a low titer (1:40 titer with vaccines for smallpox, polio, Japanese encephalitis [74]), and the associated functional properties are not well known [75]. Mucosal IgG has been associated with the protection of structure a research paper several vaccines (e.g., Hib glycoconjugates, meningococcal conjugates, pneumococcal conjugates, polio (Sabin)); whereas mucosal IgA has been associated with the protection of a smaller number of vaccines (influenza intranasal, polio Sabin, rotavirus) (reviewed in [73]). For now, a decreased risk of infection by HIV-1 vaccination is associated with antigen-specific serum antibody responses, with some evidence that certain vaccine-induced cellular responses may impact virus replication and that host genetics may modulate vaccine immunity.

Analyses of the essay thesis correlates of HIV-1 infection risk have demonstrated that there are intersections among different analyses (involving immune responses, virus and host genetics), thus providing the areas of highest information content for understanding the potential mechanisms of a research protective immunity. Identification and essay rigorous characterization of these areas of overlap are likely to a research paper, be transformative for the next phase of HIV-1 vaccine design and evaluation. We thank Peter Gilbert, Jerome Kim, Nelson Michael and a2 english critical coursework Julie McElrath for structure paper, insightful discussion in the preparation of this manuscript. The authors acknowledge funding from the HIV-1 Vaccine Trials Network (HVTN) (5U01 AI46725-05), the Duke University Center for AIDS Research (CFAR) Grant (P30 AI 64518), the Center for HIV/AIDS Vaccine Immunology-Immunogen Discovery (CHAVI-ID), National Institutes of Health/National Institute of Allergy and Infectious Diseases/ Division of AIDS and Bill and Melinda Gates Foundation Grants (1033098, 1032144, 1040758).

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