How To Write A Call To Action With 54 Words + 6 Examples

How To Write A Call To Action With 54 Words + 6 Examples All of your content marketing success hinges on the  bounce and the  conversion. The two exist on a seesaw, the dreaded â€Å"bounce† on one side and â€Å"conversion† on the other. And what is the fulcrum at the center of the two that will determine which way it will tip?  Your call to action- aka  your  marketing CTA. Your content marketing lives and dies based on the success of your CTA. It determines if people take your content and bounce out on their merry way, never to return, or if they leap for the bait and go further into your  sales funnel. Knowing how to write a call to action that  hooks your readers interest is key to your contents survival. Know how to write a #CallToAction? Here are 54 words and 6 examples thatll unlock your #creativity. What Is A Call To Action? A call to action might be as simple as asking someone to: Sign up for your email newsletter. Download  free resources in exchange for an email address. Buy an ebook,  coaching service, or your product. A call to action might be at  the beginning  of a blog post, at the end of a blog post, or sprinkled throughout a landing page. As a marketer, you use a call to action  in any medium- videos, advertisements, blog posts, landing pages, and even social messages. Whatever form it takes, and wherever it appears in your content, a call to action is you providing your reader with some form of actionable task  and usually appears as a button, link within text, or an image of some kind. Whatever form it takes, a call to action  provides your reader with an actionable task. Content marketing is creating content, but with a purpose. All content has the purpose of establishing your expertise and being helpful to your reader so that they come back to you for more.  And of course, content has the purpose of bringing in new readers through  search engines. So  you must include purposeful elements in your content that specifically ask your reader to do something tangible instead of merely feeling warm fuzzies about your brand. You must regularly ask your reader to do something. A call to action not only gets your reader to do something that commits her further to what you are ultimately selling,  it's also something you  can measure. And, if you can  measure  it, you  can test, tweak, and change  it  so you learn more about your audience. Without a call to action, you're wasting your best efforts. Without a call to action, you’re wasting your best efforts and goodwill on readers who probably would take action and who probably would buy but you've  never pushed them to. Without writing a call to action, the most you’ll know about your reader is hits, page views, and bounces. You’ll spread lots of goodwill. But you’ll never make a sale. Recommended Reading: Here’s How You Can Double Your Blog Conversion Rate Right Now How To Write A Call To Action Using Exclusivity And The Undeniable FOMO When it comes to a powerful motivator for your call to action, FOMO, or the fear of missing out, is hard to beat. This is about exclusivity, which generally works in two ways: Only some get in.  By only letting a few get in, you suggest that those who do are lucky, should be thankful, are special, are deserving- anything of this nature. This is about status, namely who’s in and who’s out. In order for this to work, you have to make something amazing enough that people want  to be in on it. Anyone gets in, but with restrictions. Think of data rights management or DRM controls on ebooks and music. The product is available to anyone, but you need specific devices, tools, or access methods to use it. In this way, it's exclusive because you control how people use it and how they  can share or spread it. That fear of missing out taps into several human emotions (some of which we’d rather not admit to). It’s more than just fear because that fear is based in something else: Panic: â€Å"If I miss out, I’ll never know if this could have changed my life!† Greed: â€Å"I have to have everything.† Comparison: â€Å"I don’t want to be the only person without this!† Curiosity: â€Å"Could this possibly be as amazing as they describe?† Pride: â€Å"I got in and you didn’t. Ha ha.† Most of us are almost compulsively driven by these emotions. When you write a call to action, you must tap into these kinds of emotion- the ones that are so connected to exclusivity- because they're what drive people to act when it comes to selling. And when you’re selling something people don’t need  (i.e. food, water, shelter, new washer) and are instead trying to create a want (i.e. you just bought a new shirt, but why not get a few more?), you have to find another motivator for them to part with their money (or their email address). And, oddly, the fear and subsequent emotions that are tied into exclusivity are good ones to use. How To Use Exclusivity And FOMO In Your #CallToAction To Boost ConversionsHow To Put Exclusivity To Work In Your Call To Action Exclusivity rides on one main idea:  If you don’t do something now, you’ll never be able to do it again. You can hint at exclusivity through the words in your call to action. Think of any word or phrase that suggests now: Last chance Limited supply Only a few left Ends tomorrow Limited time only One-time offer Expires soon Urgent Deadline You get the idea. With FOMO, there isn’t time to think. The language all points to action immediately. So Call now  is much better than Call anytime. Recommended Reading: 3 Unusual Ways To Create Exclusive Content That Attracts A Crowd How To Write A Call To Action Using Hope As A Motivation Fear isn’t the only way to get people to act, though it's one of the most powerful. Hope can do the trick, too. First you need to create a sense of desperation. Illustrate just how big a problem  your readers have, and the hope to change it will suddenly make sense. While driving around town and checking errands off of my to-do list this past weekend, I took note of the billboards and signs outside of the stores and restaurants. I said to my friend, who was with me, that there was absolutely nothing that I needed, yet here I was, buying stuff. â€Å"I wouldn’t be dissatisfied with my life and possessions if they didn’t tell me it ought to be so,† I said, a bit annoyed. â€Å"You can’t sell to people who aren’t dissatisfied,† my friend replied. With dissatisfaction comes hope. Or it should, if you’re writing  your call to action correctly. How To Leverage  Hope With  Your Call To Action To tap into hope, you first must suggest hopelessness. By that, I mean you must show the reader that there is a problem, it’s a serious one, and they have it. And then you provide the solution and the hope. â€Å"You’ve tried everything to lose weight, but nothing worked,† is easily countered with, â€Å"Try this safe and proven method that returns results every time, risk free!† In this call to action example, you assure the reader by using the words â€Å"safe† and â€Å"risk free†, and give them hope by suggesting it's â€Å"proven† and â€Å"returns results†. Big problem. Big hope. Once you’ve established this pattern, end with your simplified call to action. â€Å"Start  now and lose 10 pounds in your first month.† Recommended Reading: How To Write The Best Copy Ever With Advice From Junk Mail So ... What Are Those Call To Action Words That Get People To Act? Is it possible that certain words get more conversions than others? It's a good question that has some research behind it- to an extent. So I took a look at five well-researched articles  to pull together a  big  list of call to action words that will help you get more email signups, trials, and sales. This data comes from five  researched articles, and even includes some words that have helped get as much as a  27% conversion rate from our own calls to action: Backlinko's research on building email lists Sprout Social's research on call to action phrases WishPond's research on words to use in call to action buttons Unbounce's research on call to action buttons Unbounce's research on conversion rate optimization 's research from our own landing pages and blog posts Now, many of these sources suggest  testing your own calls to action on your blog and website to see what words perform best with your audience. That is great advice you can put into practice  by using a tool like  Visual Website Optimizer. Start simple: Write two powerful calls to action you will A/B test against one another. Set up the test with Visual Website Optimizer. Give it a week and analyze the data. The results may surprise you, as they've done with us at . 54 Powerful Words To Test In Your #CTAs6 Call To Action Examples To Help You Write  Better CTAs Than Ever Call to action examples are everywhere, so where would you  start? I figured we'd take a look at a few  of the top companies in the world according to Fortune's 500 list (there has to be a reason they're so successful, right?), then dive in to some specific examples that are a bit more content marketing related. Let's do this. 1. Learn More With Apple Like the simplicity of their products, Apple  keeps their calls to action short, clean, and to the point. Learn more and Trade up to a new iPhone are unmistakable in letting Apple's  audience know exactly what they'll see after they click through the call to action. Apple  also doesn't clutter the design: There are clearly only two options to help their users focus on making a decision to click quickly and easily. Lesson Learned: Be clear and concise,  and position your call to action as the  obvious next step. 2. Save With CVS While the design is super busy compared to Apple's example (do I click on Save with Our App, App Store, Google Play, or on the phone itself?!), CVS focuses on the value proposition  in their call to action. Save with Our App  focuses CVS'  audience on the perceived benefit of using the tool, which connects into hope to resolve the dissatisfaction of spending so much money. The headline here- Unlock  extra savings  with app-only deals!- also suggests exclusivity, that you can only experience those benefits if you get the app. They also include the word exclusive in the description, which is a powerful motivator for the fear of missing out. Lesson Learned: Keep your design clean so your users know where to click. Brainstorm the value proposition to answer your readers' inherent question, "What's in it for me?" and tie that into your call to action. 3. Rely On The Visual Like Amazon Who said  you had to write a call to action? The behemoth Amazon focuses on compelling imagery to entice action. In this example, there is really no  traditional call to action that usually begins with a verb. However, Amazon does rock some powerful words with the visual: Now and Limited time offer. Talk about inducing urgency and appealing to the fear of missing out. The visual tells the rest of the story, helping  Amazon shoppers envision themselves in front of an amazing TV with a bowl of popcorn and a couple glasses  of champaign. Would you like to chillax like that, too? Heck yes, you would. And that's why the visual creates a powerful  call to action. It's noteworthy to mention this: Amazon doesn't even show the product they're selling (other than the remote) and focuses the visual mostly  on the experience  you'll feel when you click through the call to action to purchase it.  People don't buy products, they buy experiences. And that's why Amazon crushed  it with this call to action example. Lesson Learned: Literally show the pleasurable  outcome your audience will experience if they just click through your call to action. People don't buy products, they buy experiences. Include the experience in your #CTA.4. Code School Shows Calls To Action Don't Have To Be Super Formal Wondering how to apply calls to action into your blog posts? Write a post that helps your readers do something without you, then  when you can  help them  do it even better, let them know. Code School helps people learn how to code. So when they wrote a post called Why Python? that covered reasons to learn and use the language, they ended the post with a call to action to learn more through the courses Code School offers. The call to action appears as the last paragraph in the blog post, and invites readers to Check out a couple new courses. It's simple, informal, yet informational to teach their audience about the possibilities Code School offers to help their readers  improve. Lesson Learned:  Write a blog post that connects into your product or service. Then end the post  with an informal call to action to work  with you to resolve the problems you just outlined in your post and link to content that introduces  your offering. 5. Wistia Embeds Calls To Action In Videos Do you embed videos in your blog posts? With Wistia, you can add  a Turnstile into your  video  to collect email leads during video play, and you can also include a written call to action and link to related content at the end of your video. Turns out, you can also write a call to action right into your YouTube videos, too, if you're not a Wistia user. Anyway,  this call to action example shows up at the end of a video that explains one element about making post-product process simpler- something Wistia's audience cares a lot about. Wistia  then links to more information on the topic to help their viewers learn more about post-production- which is a part of the video process Wistia as a tool  can help make more efficient. The video appears in lieu of a traditional blog post header graphic. That  means Wistia's audience sees a video right away (which also likely boosts on page time and engagement quite a bit), Wistia shows their quirky brand personality, and they display a  strong call to action immediately. This is a great example of leading an audience deeper into the funnel  from inbound marketing  to demand generation content that positions Wistia as the solution to the problem. Brilliant. Lesson Learned: Strategically think about the next step to bring new users from inbound marketing into demand generation content that positions your product or service as the answer to a problem your audience is facing. Write  calls to action for all of your videos. 6. Create Content-Specific  Calls To Action Like Backlinko Have you seen blog posts that offer something free in exchange for your email address? Somewhere along the lines, marketers started calling those things content upgrades. And they are a super smart way to include a call to action in every blog post to turn the traffic you get into email subscribers. In this call to action example from Backlinko, Brian Dean includes a written CTA in the introduction of his blog post. That is brilliant  because a majority of your audience will read the first 100 words of your post, then maybe skim the rest. Anyway, this call to action is very smart because it relates specifically to the content at hand and not just a generic give-me-your-email-address kinda CTA. Brian relates directly to the challenge his readers want to resolve by writing Get More Email Subscribers, then he uses a powerful word with download. He also promises that what his readers will get behind the call to action will help them quickly execute the 17 strategies from this post, which is very important for the Backlinko blog in particular  because Brian  creates such long-form content. This way, he captures the interest of the too-long-didn't-read (TLDR) audience while also turning them into email subscribers. To top it off, Brian targets another call to action for the 20% of his audience that makes it through the entire blog post to the very end. This time, he lets the visual speak for itself with the power word download, and  includes the emotional word free to describe his  content upgrade. Lesson Learned: Include a  relevant content upgrade at the beginning and end of your blog posts to convert  traffic into email subscribers.  Use your call to action to appeal to the reasons your audience is already interested in the content, packaging a condensed and actionable guide behind the clickthrough. 6 #CallToAction Examples That Will Unlock Your #Creativity! #bloggingPut It All Together: How To Write A  Call To Action With A Compelling  Structure No matter what motivation you use (fear or hope), there are some common ways  that you should use when structuring your call to action. 1. Start with verbs. Verbs are the action words that make it clear to readers what you want them to do. Instead of saying, â€Å"Ready to get started?† simplify it to â€Å"Get started now† or â€Å"Start saving now†. Some verbs are stronger than others. This has to do with the cacophony of the word  (hard K, G, D  sounds) coupled with  the strength of the action suggested in context. The word â€Å"buy† feels stronger and more urgent than â€Å"purchase†. â€Å"Get your copy† is more cacophonous and powerful than â€Å"Download your ebook†. Starting with verbs means starting phrases and sentences, sure, but also the placement of links and call to action buttons. They come first (or nearly first) and should be prominent. Buttons to buy or sign-up should be above the fold, no scrolling required. They should be before the long chunk of explanatory text. Verbs are words of action, and that’s exactly what you want from your readers.2. Take it easy on filler words. Adverbs and adjectives can get in the way of the action you want your readers to take. You should have already done the work of convincing people to take the action before presenting your call to action  to them through blog post or landing page copy. Try your best to avoid words ending in -ly. â€Å"Click here† is better than â€Å"Click here quickly.† There are exceptions, though.  For example, you might hint at exclusivity by saying â€Å"Get your custom ebook now† instead of just â€Å"Get your ebook†. 3. Keep things simple and brief. Use words that are simple, common, and not too long. This isn’t because you think your reader isn’t smart enough to handle anything else, but because you’re trying to prod base emotions. You don’t want anything to get in the way of those simple emotions, particularly requiring readers to consider complex thought processes or scenarios which could distract them or lead them down a path of thought away  from taking action. Avoid buzzwords, jargon, and any word that feels â€Å"empty† and can be ignored by the reader. If you use enough ignorable words in your call to action, your entire call to action runs the risk of being ignored. Steps for a successful #CTA: verb  Ã‚  clarity  Ã‚  simplicity request.If you must use descriptive words, use simple and common words that are emotionally effective. This is no time to go crazy with a thesaurus and impress the world with your vocabulary. The language must not  get in the way of the emotional prodding you’re trying to achieve. Save your thinking words for your blog posts, and focus your call to action on words that are powerful persuaders. Remember simple, basic, and primal words- and not too many words overall- when you make the big request. 4. Make the request simple, too. It’s not just the language of the request that you need to make simple, but the request itself must be easy. One or two clicks to completion. As little pain as possible. If you need more information than a name and email, try to break it up so you capture that email first, get the user into the purchase or into their account, and then collect the rest later. The reason is that you don’t want to give your reader time to change their mind. I don’t know how many times I’ve been at a big box store and, because of too few checkout lanes available, seen people abandon full carts in line and walk out the door. It’s the same with your call to action. The more complicated you make it for readers to complete it, the more likely they’ll find a way to change their mind or forget the driving reason you just convinced them they needed to take action. â€Å"Buy now!† doesn’t feel like â€Å"now† if you make them fill out lots of information, answer a small survey, and click on a reply email to verify. That feels like buying later, and it brings into question the urgency you insisted was necessary to get a reader to take action.

Course work questions 10-11-12 Coursework Example | Topics and Well Written Essays - 1000 words

Course work questions 10-11-12 - Coursework Example As a teacher, encourage correction from both the person and the group incase a mistake happens. Participate in correcting students where they go wrong by first explaining why that is wrong then giving the correct answer. How would you introduce your students to the culture of an English-speaking country? Please describe an activity, which would help them learn more about the country and increase their cultural sensitivity. You may use one of the activities mentioned in the module, (Polls, Excursions, films, Q&A, Guest Speaker) see upload from module. I will use guest speakers who have integrated into the new culture. Before every session, I will first alert my students that we will have a guest speaker in the class to help us understand the new culture. Being their teacher, encouraging students to prepare questions on the culture will be helpful to avoid cases of no response to anything presented by the speaker. I will talk with the guest speaker on the issues I will need him/her to handle concerning the cultural differences in my class before the session. Once the guest speaker is in class, I will introduce him/her and once again encourage students to ask as many questions as they can to the guest speaker. Later on, we will have an open session where students will give feedback on the importance of having the speaker and how well the session has helped them understand the culture of an English speaking

Marketing management MMP 2 Essay Example | Topics and Well Written Essays - 2000 words

Marketing management MMP 2 - Essay Example The projected sales of the company for the year 2008 is followed by the objectives for the year and marketing strategy, the means to achieve the projected sales and bottom line numbers. Magic Carpets Inc is a 15-year-old company specializing in the carpet cleaning and is among the top four carpet-cleaning companies based in California. The company decides to get proactive in handling the growing competition in the field especially from the large players in the janitorial services who have a pan American presence and a few multi nationals who are planning to venture into the market in a big way. Magic Carpet believes that it is high time it consolidates is business, capitalizes on its strengths and makes a few smart moves to increase market share and wallet share. The entire approach of Magic Carpet is built around establishing the brand, building loyalty, making differential pricing strategies, acquiring new markets, building new channels of business, nurturing business partners and coming out with innovative service offerings to build value to its services. The action plans are worked out in accordance to the 4P framework – Product, Pricing, Place and Promotion. (Kotler Philip, 2004, Marketing Management) The 4P’s are applied across two broad market segments – existing markets and new markets. The following are the major action plans made to achieve the business objectives of ’08: For Magic Carpets, the elaborate business plans made for 2008 necessitated elaborate controls. With different process owners involved at various stages of execution of the action plans, there was a definite need to build adequate checks and balances at every stage of the plans to ensure successful completion to derive the necessary results. It is not important for Magic Carpets to achieve the results only. The means by which the result is achieved is equally important. Hence, process adherence is of paramount importance to successful

Impacts on Quality of Life Among School Children

Impacts on Quality of Life Among School Children PAVITRA A/P INDRAJOTHY INTRODUCTION It is the truth to say that children are our future and if that is true, more time and energy should be put into the beginning stages of each childs life. The early years of a child’s life are the most important in terms of their general well being, their emotional and social development, and their physical, intellectual and emotional growth. Children develop at different paces and a very high proportion of what they learn takes place in the first five to seven years of life. What happens in the home is extremely important to development in early childhood. There are things that can and should be done to help each child live a life that is productive and beneficial to those that they will interact with in their future. There are many aspects in todays world that make the quality of life of school children better and meaningful. For instance of aspects that can affect quality of life are obesity, bullying, lifestyle, emotional problems, divorce, socio cultural home environment, child abuse, oral health and back pain. First and foremost, there are reports saying that obese children demonstrate more negative self-perceptions, decreased self-worth, increased behavioural problems, lower self-esteem and lower body esteem and perceived cognitive ability. However, because of methodological limitations, definitions of obesity that differ with current recommendations and with non-validated measurements of health related quality of life, it is difficult to generalize the conclusions of these studies (Friedlander et al., 2003). According to research done by Nathan Woon in year 1981, Peninsular Malaysia is a multi-racial country of 12 million people, 41% of whom are less than 14 years of age (Vital Statistics Peninsular Malaysia 1975-Department of Statistics, Malaysia). The population consists of three main ethnic groups: the Malays (53%), the Chinese (35%) and the Indians (11%) (Vital Statistics, Peninsular Malaysia, 1975). Whilst in recent years, there has been an increasing tendency for the various ethnic groups to intermingle and to respect and understand the various cultures and traditions that coexist within the country, in the specific area of child rearing, as with marriages, deep-rooted traditions and customs continue to remain a crucial factor and probably contribute, to some extent, to the pattern and incidence of child abuse in the various races. Bullying has been conceptualized globally as acting in any way that threatens or hurts someone less powerful. Furthermore, bullying also can be distinguished among the different way in which aggressive acts are performed. There are different types of bullying which is physical, verbal, or indirect bullying. Furthermore, distinctions among aggressor’s possible aims can also be made. For instance, bullying physically to hurt the victim or for damage the victim’s relationship with others (Rigby, 2003). Next, parental separation is usually the first major and drastic change in a child’s life. This distressing event drastically alters the family’s future, bringing to a sense of loss due to interference in daily routines as well as absence of daily contact with both parents (Eymann et al., 2009). Lastly, obesity, dental caries and periodontal diseases are among major public health concerns which may affect children’s growth and development. Sedentary lifestyle and unhealthy eating habits could lead to poor oral hygiene statuses and increase the tendency to develop periodontitis and obesity at young ages (Anand et al., 2014). DISCUSSION OBESITY Worldwide estimates of childhood obesity are as high as 43 million, and rates continue to increase each year. Childhood obesity is a growing problem in the present era and it causes serious consequences in the later years (Anand et al., 2014). According to Friedlander et al., childhood obesity is a major public health problem with increasing prevalence; approximately 30% of US school-aged children are defined as overweight or at risk for overweight. Anand and Friedlander states that overweight children are more likely to suffer from a range of chronic health problem such as cardiovascular disease, diabetes mellitus, hypertension, sleep apnoea and asthma. They also concluded that children who are overweight are more likely to become overweight adults. There was a study conducted by Friedlander et al. among 371 students aged 8-10 years old excluding those who had sleep apnoea or who were born prematurely. Based on BMI categories of the study, 17.5% of the children were overweight, 12.4% were at risk for overweight, 8.1% were underweight, and 62.0% were of normal weight. Moreover, children who are overweight have a 2 to 4 times increased odds of having low scores for physiological health, self-esteem, and physical functioning. Similarly, children who are at risk for being overweight have higher odds of having low physical functioning scores. Furthermore, lower self-esteem also was observed in children who are underweight. Children with higher BMIs are more likely than normal weight children to be perceived by their parents as having feelings of anxiety and depression, of exhibiting aggressive or immature behaviour, as having role limitations in their schoolwork and social activities, and as having low self-esteem. And, their parents also report higher levels of emotional distress and little personal time because of their children’s health or behaviour (Friedlander et al., 2003). As a suggestion, if the child or teenager is overweight, further weight gain can be prevented. Parents can help their children in maintaining their weights in the healthy range. In infancy, breastfeeding and delaying introduction of solid foods may help in preventing obesity. In early childhood, children should be given healthy, low-fat snacks and they should be allowed to take part in moderate-vigorous physical activity every day. Older children can be taught to select healthy, nutritious foods and to develop good exercise habits (Anand et al., 2014). BULLYING Bullying invariably involve an imbalance of power in which the aggressor is more powerful than the victim. Moreover, bullying occurs when there is conflict between people of unequally power (Rigby, 2003). As identified by Rigby in year 2003, four categories of negative health conditions are low psychological well-being, poor social adjustment, psychological distress and physical unwellness. Low psychological well-being includes states of mind that are generally considered unpleasant but not acutely distressing, such as general unhappiness, low self-esteem, and feelings of anger and sadness. Next, poor social adjustment normally includes feeling of loathing toward one’s social environment, evident through expressed dislike for workplace or school, manifest loneliness, segregation and absenteeism. Moreover, psychological distress is considered more serious than the low psychological well-being and poor social adjustment categories and comprises high levels of depression, anxiety and suicidal thinking. Last but not least, physical unwellness can determine by precise signs of physical disorder, evident in medically diagnosed illness. Furthermore, psychosomatic symptoms also can be incorporated in this category. Various strategies or treatment should be considered and give a thought to reduce the chances of a child’s further severe involvement in bullying that can worsen the child’s condition. The approach include helping victimized children to evolve self-protective assertiveness skills and working therapeutically with bullying children to create a greater awareness of the repercussion of their antisocial behaviour (Rigby, 2003). LIFESTYLE Television is still the most widely-viewed screen worldwide. Globally, people are spending more time sitting at work and at home, and there’s mounting evidence that this â€Å"sit time† is a major contributor to the obesity epidemic. Sedentary activities not only TV watching, but also working at desk jobs, using computers, playing video games and driving cars. Increasingly, there’s evidence that watching TV and, especially, watching junk food ads on TV promotes obesity by changing mainly as to what and how much people eat (Anand et al., 2014). Too much television viewing among children has been linked with inadequate study patterns. Inappropriate television-viewing has been linked to erratic sleep/wake schedules and poor sleep quality, violent or aggressive behaviour, substance use, sexual activity resulting in decreased school performance or even school drop-out (Karande Kulkarni, 2005). EMOTIONAL PROBLEMS Conditions which cause emotional problems in children such as chronic neglect, sexual abuse, parents getting divorced or losing a sibling might cause long term distress resulting in academic underachievement. Besides that, children can face severe emotional upheavals during the treatment of chronic health impairments such as asthma, cancer, cerebral palsy, congenital heart disease, diabetes mellitus, epilepsy, hemophilia, rheumatic diseases, or thallasemia, resulting in low self-esteem and loss of motivation to study (Karande Kulkarni, 2005). As recommended by Karande Kulkarni in year 2005, children with emotional problems need counselling sessions with a child psychologist or a child psychiatrist. Depending on the severity, at times, appropriate medications (anxiolytics, antidepressants) may be needed. DIVORCE As emphasizes by Eymann et al. in year 2009, divorce is becoming increasingly common in our society, seriously affecting all parties involved, especially children. There were studies conducted in USA and in Europe and roughly calculated that 30 to 50% of children will suffer the effect from a divorce, almost same rates have been reported in Argentina. Eymann et al. conducted a study at the Hospital Italiano, Argentina. A total of 330 families were invited to take part and only 313 completed the questionnaire; 160 from married family and 153 from divorced family. They defined divorced family (DF) as parents who choose to stay at separate residence regardless of legal marital status while married family (MF) as parents who choose to share residence with their children regardless of legal marital status. The aim of this study was to evaluate psychological quality of life among school aged children of divorced parents by using Child Health Questionnaire – Parent Form 50 (CHQ-PF 50). The results showed that CHQ-PF 50 scores were higher in children of MF than in those of DF for various domains, such as self-esteem, time impact on family, mental health, behaviour, social-emotional functioning and behaviour, and lastly the psychosocial summary score joining all these factors. As proved in the results, boys had poorer psychosocial quality of life than girls. This was explained perhaps by the effects of less daily father-child connection on gender-specific identity traits. Furthermore, it is well known that separated mothers who have legal child custody usually suffer a greater slope in their standard of living. Norton Glick and Duncan et al. have clearly proved the unfavourable economic repercussion of divorce on child well-being. In this study, maternal remarriage didn’t appear to influence quality of life of child. Maternal remarriage could impact children who must readapt to new family basis and dynamics. However, a new partner perhaps could provide additional financial support since parental separation usually involves a fall in family income. Moreover, the new couple may construct an improved marital relationship, hence could minimize the distressing emotional burden on children and making the stepfather as a positive and ideal role model (Eymann et al., 2009). Lastly, in this study, separation by mutual agreement because of higher maternal education or maternal university education level and longer the length of time passed since separation has inclined psychosocial quality of life scores, while scores declined in boys and for each one-year increment of maternal age. Eymann et al. proves that higher maternal educational level could give positive impact on psychosocial quality of life, which may be explained as a consequence of the skills build through education years, allowing better and organized control of the marital problems and crisis and greater agreement on conflict issues (Eymann et al., 2009). CHILD ABUSE A prospective study was made of all cases of child abuse seen at the University Hospital, between July 1978 and June 1980. A total of 19 cases, 16 of physical and 3 of sexual abuse were diagnosed during this period. The relatively high (32%) incidence of child abuse amongst Malay families was quite surprised, as Malay children are generally much loved and pampered. Nathan and Woon suspect that the increasing incidence of child abuse amongst the Malays is probably related to their urban migration in recent years with the resultant breakdown of the extended family system and the need to cope with the pressures of life in an unfamiliar urban setting (Nathan Woon, 1981). The commonest injuries found on admission were bruises and injuries to the face and scalp. The majority of the abusers in our series were parents, the ratio of father to mother being. Both social and personal factors in the life of the abuser seem to contribute to the act of child abuse. Marital disharmony appears to feature prominently amongst the interpersonal problems, while two abusers gave a history of having been abused physically during their early childhood (Nathan Woon, 1981). SOCIOCULTURAL HOME ENVIRONMENT It has been recognized that children from poor socioeconomic status families have higher chances of poor school performance. Malnutrition due to poverty coupled with low education and status of parents adversely affect their cognitive development. Such children also have higher chances of experiencing, right from their pre-school years, parental attitudes which do not motivate them to study and an unsatisfactory home environment which does not encourage learning (witnessing domestic violence, family stressors, and adverse life events). Alleviation of hunger, by providing one balanced meal in school is one of the mechanisms to improve academic achievement in undernourished low-income elementary school children (Karande Kulkarni, 2005). Besides that, many of these disadvantaged children are studying in English medium schools as their parents believe that this would help them progress in life. These children face the added burden of language barrier, namely, they are not conversant in English as they came from non-English speaking families, which leads to poor school performance or even school failure. Parents of children with language barrier should be counselled to educate their children in their own language medium schools or to attend a facility for language stimulation if that is available and affordable (Karande Kulkarni, 2005). ORAL HEALTH In the research carried out by Anand et al. in year 2014, they noticed that poor oral hygiene statuses in children who spent more time watching television, playing video games, as their consumption of soft drinks and fast foods was more and due to lack of adequate exercise. BACK PAIN Musculoskeletal pain is exceedingly common in young adults. A questionnaire survey was carried out by Brattberg in year 1994 among children 8, 11, 13 and 17 years old. The prevalence of back pain and headaches among 1,245 Swedish school children was studied. 29% of the students reported back pain and 48% headache. In all age groups studied, both back pain and headaches were more common among girls than boys. Girls also reported more frequent symptoms than boys (Brattberg, 1994). CONCLUSION Children who are at risk for or who are overweight having odds 2 to 4 times greater for lower health related quality of life scores for physiological health, self esteem and physical functioning. Parents of children who are overweight also were more likely to evidence higher levels of emotional distress because of their children’s health. Being relatively underweight also was associated with lower self-esteem (Friedlander et al., 2003). Ken Rigby in year 2003 concluded that being victimized by peers is notably related to relatively low levels of psychological well-being and social adjustment and lastly to high levels of psychological distress and unfavourable physical health symptoms. Eymann et al. in year 2009 concluded that children’s psychosocial quality of life was affected by divorce. Psychosocial quality of life improved when parents had divorced by mutual agreement, the mother had a university education, when more time had elapsed since separation and was poorer in boys and in children of older mothers. And, quality of life can be improved as more time elapses after the divorce, this is because may be it can be interpreted as a process of accepting the family modification. And lastly, poor oral hygiene was observed in children who spent more time in leisure activities and that lifestyle factors could make an impact on oral hygiene and periodontal health. A need exists for addressing obesity, oral health and nutrition, jointly in health promotion strategies, to improve well-being of children and also to empower good life-style factors (Anand et al., 2014).

Japanese Occupation in South-east Asia Essays -- Pacific War Japanese

Japanese Occupation in South-east Asia Table of Contents Chapter 1: Introduction Background  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 3 Thesis  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 3 Research questions  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 3 Rationale  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 3 Methodology  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 4 Chapter 2: Literature Review  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 5 Primary sources  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 5 Secondary sources  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 6 Chapter 3: Research Methodology  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 8 Procedure  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 8 Types of sources  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 8 Compiling and presenting the data  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 8 Chapter 4: Results and findings  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 10 Background information  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 10 Conflict between Japan and United States  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 11 The Japanese Occupation (1942 – 1945)  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 11 Chapter 5: Discussion and interpretation  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 24 The causes or motives of Japanese Occupation  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 24 The effect of Japanese Occupation  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 25 Propaganda  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 26 Chapter 6: Conclusion  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 28 Acknowledgements  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Pg. 29 Chapter 1: Introduction Background:   Ã‚  Ã‚  Ã‚  Ã‚  The Pacific War opened on 7 December 1941, and Japanese troops started invading other Asian countries. Singapore, which was a colony of Britain at that time, fell to the Japanese on 15 February 1942. Since then, the people in South-east Asia had great sufferings. During the Japanese Occupation, many people suffered and some even died. Finally, the Japanese surrendered after the bombing of Nagasaki and Hiroshima, and the nightmare was finally over. Thesis:   Ã‚  Ã‚  Ã‚  Ã‚  War does more harm than good. Research questions: †¢Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  What are the main causes & motives of the Japanese Occupation †¢Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  What are the main effects of Japanese Occupation to the people at that time? †¢Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  What are the instruments... ...lusion The people under the Occupation suffered, and the people in Japan also suffered after the bombing of Nagasaki and Hiroshima. We learned that war does not do any party good, as we can see from the end of the war. We should learn our lessons from this event, and from the causes we can learn how to prevent it. Therefore, we should place our attention on the recent US and Iraq war, and stand on the side of peacefulness. We should also educate ourselves to differentiate between propaganda and the reality. Lastly, I had proven my theory, â€Å"War does more harm than good.† Acknowledgements http://www.iwm.org.uk/upload/package/25/pearl_harbour/hb_rise_of_japan.htm http://www.iwm.org.uk/upload/package/25/pearl_harbour/hb_ww1.htm http://www.iwm.org.uk/upload/package/25/pearl_harbour/hb_us_lead.htm http://www.iwm.org.uk/upload/package/25/pearl_harbour/hb_j_agg.htm http://www.iwm.org.uk/upload/package/25/pearl_harbour/hb_count.htm http://www.japan-guide.com/e/e2130.html http://www.japan-guide.com/e/e2129.html http://www.diggerhistory.info/pages-posters/jap.htm http://www.s1942.org.sg/ Understanding our past, Singapore from Colony to Nation - Federal Publications

GFP protein

Green Florescent Protein, abbreviated as GFP, is a protein composed of 238 amino acids that is commonly found in mnemiopsis, comb Jelly. It has a major wavelength at 396 nm and a minor one at 475 nm. GFP is what gives mnemiopsis their bright green florescent glow. ultraviolet light, or blue light, is necessary to see the florescent glow of this protein. GFP is an irregular protein because It Is highly resistant to denaturation by temperature and PH. It can survive In temperatures up to 98 degrees and has a pH of 12. 2 due to Its complex exterior, called the beta barrel. At an pH higher than 12. It denatures. It also has an Isoelectric point at 5. 3. The peripheral beta barrel cannot be digested or broken apart by protease because of the strong bonds holding It together. The beta barrel protects the chromophore, which Is the substance which gives GFP Its green glow. When CFP Is extracted from the plasmid of an E. Coll or from a Jellyfish, It contains an array different contaminants ma king it difficult for scientist to do experiments with GFP. A procedure in purifying GFP from a crude cell extract to nearly 100% GFP so that it can be analyzed and used in scientific experiments and research is necessary.The goal is to ptimize each protocol used to purify crude GFP. Methods Ammonium Sulfate Precipitation To purify the crude samples of GFP, the ion exchange method separates substances inside the test tube by similar charge. A sample of crude GFP of 7. 5 mL in a plastic tube was used for the experiment. Knowing that 43. 6 grams of ammonium sulfate in a 100 mL solution yields a 70% percent saturated solution, the proportion 43. 6g 11 00 mL=x/7. 5 mL was used to determine that 3. 27 grams of ammonium sulfate needs to be added to the experimental sample. After adding the ammonium sulfate, the solution was stirred gently to prevent frothing.Once most of the solution is transferred, the tube was placed on a triple beam balance along with another tube that went through the same process. The centrifuge was set at 15,000 rpms for 15 minutes so that the hydrophobic materials will separate and become the supernatant while the GFP pellet will remain behind. Once the 15 minutes elapsed, a new pipette was used to remove the supernatant, leaving behind the pellet of GFP and hydrophilic contaminates. To remove the hydrophilic substances, 5 mL of 4 molar ammonium sulfate and 15 mL of 10 mL tris at a p of 8 was added Into the oak ridge entrifuge test tube.The solution Is then put Into the centrifuge at 15,000 rpm for 15 minutes again. Once 15 minutes has passed, the supernatant, containing the GFP, was removed by a pipette and put In a microfuge. Hydrophobic Interaction Hgure yaropnoDlc Interactlon set up One molar ammonium sulfate was added to the column to wash the sample. Adding 1 molar ammonium sulfate washes the sample because a high salt concentration increases hydrophobicity of the GFP and the buffer, causing most of the GFP to be at the very top of the column. Substances that are hydrophilic get flushed out of the olumn while the more hydrophobic substances stay in the container.After the column has been eluted with 1 molar ammonium sulfate, the tris buffer is added to the ammonium sulfate to dilute it into . 5 molar ammonium sulfate. The volume of 1 molar ammonium sulfate inside the oak ridge centrifuge test tube is the volume of the tris buffer that will be added. After the column chromatography has been flushed with . 5 molar ammonium sulfate, more hydrophobic substances will be flushed out since the hydrophobicity of the tris buffer and the GFP has decreased. This causes the GFP to spread out in the column. Finally the amount of . 5 molar ammonium sulfate is diluted with tris buffer to . 5 molar ammonium which should cause most of the GFP to be flushed out of the column along with other substances that are very hydrophobic. While this experiment is going on the liquid that comes out of the column is collected in multiple test tubes. These test tubes contain GFP and other contaminants. The solutions are than read by a spectrophotometer. Each test tube will be tested by the spectrophotometer so that a graph can be made. Anion Exchange Figure 2: Siphon Bridge set up for Anion Exchange Figure 3: Centricon Test Tube In order to use anion exchange, the starting condition of the sample needs to be in a low salt solution.However after the GFP had gone through hydrophobic interaction, it was in a high salt solution. Before facing this dilemma, the fractions were pooled by centricon which decreases the overall sample volume by removing some buffer and salt solution. This greatly increases the GFP concentration in the samples. The fractions are placed in the centricon and then into a centrifuge for 25 minutes at 3,000 rpm to be separated by size. The large proteins stay in the entricon while buffer and salt solution goes into the plunger. To reduce the concentration of salt in the GFP sample, the sample is diluted 10 folds.Since the amount of GFP that was restored was 18 mL, 162 mL of tris buffer needed to be added. The diluted GFP is then put in the chromatography container, containing positively charged DEAE which is attracted to the GFP at a low salt concentration. After the GFP has been completely filled, the column is connected to a beaker that contains a low salt concentration. the low salt concentration beaker is connected to a high salt concentration beaker. As one drop of low salt solution goes into the chromatography column, one drop of high salt solution goes into the low salt solution.Gradually the salt concentration increases in the low salt beaker and in the column chromatography, causing GFP to spread down the container. The eluted GFP dripped out of the column chromatography to be collected in test tubes. I nree pnase partltlonlng Figure 4: Precipitate of GFP. T-butanol is one top while contaminates are on bottom GFP then went through three-phase partitioning, also known as TT P. The fractions taken after an anion exchange was 15 millilieter. Ten ml of 4 M ammonium sulfate was added to this volume to increase the salt concentration of the solution to 1. M, which is about 40% salt saturation. Twenty-five milliliters of t-butanol was added then added which was the same amount of ammonium sulfate and GFP in the container. The container was then placed in the centrifuge for ten minutes at 4600 RPM, causing the mixture to split into three layers; butanol on top, GFP in solution on the bottom, and precipitated contaminants in-between. The top layer of butanol and disk of precipitate were taken out. The volume of GFP solution was again matched in utanol and the container went into the centrifuge again. An aspirator was used to extract the GFP into a microfuge. . 6M ammonium sulfate was then added to the microfuge and the container was placed in a micro centrifuge for one minute at 13,000 RPM. Butanol and other contaminants that had not been take out previously f ormed a disc, was then taken out with an aspirator and the remaining GFP was then left in the microfuge. HPLC Figure 5: HPLC basic layout After the sample went through three phase partitioning, it was put through the High Performance Liquid Chromatography for a final purification. First liquid was put into the HPLC to clean out any previous GFP inside the loop of the HPLC and the column of the HPLC.Then, GFP in the microfuge was sucked into an injector to be put into the HPLC. Pushing the top of the injector slowly, GFP entered into a loop inside the HPLC. Once the GFP was placed in the loop, a knob was turned clockwise to the word lock. The GFP was then sent to the column where it was purified further by size through the minuscule beads. About 6,000 pounds of pressure per square inch was produced by the HPLC to push the GFP through the beads. While this was occurring, a pectrophotometer connected to the HPLC read the wavelengths of substances.Near the 396 nm wavelength, GFP was col lected in a microfuge tube. A UV light was held near tne exlt 0T Results e HPLC to measure tne amount ng sample. Graph 1: Results of the sample after HIC at a wavelength of 395 nm Graph 2: Results of the sample after HIC at a wavelength of 280 nm Graph 3: Results of the sample after HIC of the entire spectrum Seventeen test tubes were received after the HIC purification process. A blank consisting of tris buffer and ammonium sulfate was sampled in the spectrophotometer against liquid from each of the seventeen test tubes.Graph one represents the sample after HIC at a wavelength of 395 nm while graph two Results shows the results after HIC at a wavelength of 280 nm. After HIC, the fractions 12 to 16 were chosen for their purity and recovery of GFP. Graph one show the amount of GFP in each fraction number while graph two shows the total amount of protein in each fraction number. Graph three shows the spectrum of the entire sample. Graph 4: Results after Anion Exchange at a 397 nm wave length Graph 5: Results after Anion Exchange at a 280 nm wavelength Graphing 6 Thirteen test tubes were collected from the Anion Exchange purification process.This time the samples were blanked against tris buffer at 8. 0 pH and 0. 5 molar sodium chloride. Graph four shows results of the Anion Exchange at a 397 nm wavelength and graph five shows the results after Anion Exchange at a 280 nm wavelength. Once again, the graph at a 297 nm wavelength shows the amount of GFP while the graph at a 280 nm wavelength shows the amount of total protein. Graph six represents the results of the entire spectrum. The GFP peak was a lot more visible. Step Iotal sample (mL Abs (280) Total Protein Abs (397) GFP Ratio Crude sample 120 1600 . 25 At-ns042- 20 1 . 61 . 9 118 HIC 18 . 28 . 173 . 618 15 . 126 . 130 1. 03 3 Phase Partitioning . 01 n/a . 75 . 243 . 257 1. 06 Table 1: This is the overall data table. The second column shows the total volume at the start of each purification step. The following two columns are the peaks of the graphs at those wavelengths. The last column represents the ratio of GFP to the total Protein. The most desirable ratio is 1. 25. Dlscusslon The first method in purifying the crude GFP was using the ammonium sulfate precipitate. When ammonium sulfate is placed in water, it dissociates into ammonium (NH4+) and sulfate ions (S042-).Water, composed of two hydrogen ions and one oxygen ion, is a polar molecule because the oxygen has a high electronegativity. Oxygen has a greater affinity making the oxygen portion of water negative and the hydrogen portion of the water positive. The dissociated positively charged ammonium ion is allured to the negatively charged oxygen while the negatively charged sulfate ions are attracted to the dissociated positively charged hydrogen. The attraction between the ammonium sulfate and the water was so strong that the GFP and other proteins were left unoccupied, causing them to precipitate.When GFP in the 70% salt solution was placed into the centrifuge, substances such as DNA and RNA was removed because they became part of the supernatant. At a 70% salt concentration, only hydrophilic substances stay in solution while the more hydrophobic substances precipitate. When the GFP in a 25% solution of salt was placed in the centrifuge, the GFP and other substances went back into solution because there not enough water was occupied by the salt. Before the GFP is placed in the centrifuge, it must be balanced with another centrifuge with the same weight and the two containers must be placed across from one another.This is vital because the centrifuge needs to be balanced when it is rotating at an incredibly fast speed. Failure to have balanced centrifuge containers can result in a broken centrifuge and loud sounds. Also when mixing the GFP with salt, it is important not the shake the container or frothing will occur, making it difficult to transfer the solution in to an oak ridge centrifuge tube. The second p urification procedure that GFP underwent was hydrophobic interactions. During this purification, GFP binded to the non-polar Phenyl Sepharose beads because of its non-polar and hydrophobic traits.However the water in tris buffer is strong enough to separate the attraction between GFP and the Phenyl Sepharose. Therefore a high salt concentration is necessary to occupy the water so that the GFP and the Phenyl Sepharose to be attracted together. At a high salt concentration, GFP with bind easily to the Phenyl Sepharose since very little water molecules would interfere with the attraction and at a low salt concentration, GFP would not bind easily to the Phenyl Sepharose because tnere wlll De a lot 0T unoccuplea water molecules tnat wlll De aDle to InterTere wltn the GFP and Phenyl Sepharose attraction.Before the experiment, ten millimolar tris buffer at a pH of 8 was used to clean the column in order to keep the pH stable and to wash away the salt, ammonium sulfate, in the column. Remov ing the salt is vital because the buffer that once surrounds the salt will be allured to the hydrophobic benzene and to the hydrophobic patches on the GFP. Since the hydrophobic patches of the GFP are already filled, they will be flushed out, leaving mostly beads of benzene and the 10 millimolar tris buffer at a pH of 8. Once the column has been clean, it needs to be equilibrated so that the salt concentration is the same through the olumn.The step gradient used, started ata 1 molar ammonium sulfate concentration and was halved until a . 25 molar concentration to separate substances by hydrophobicity. The third purification procedure was anion exchange. In this procedure, GFP and other contaminants are separated by charge. The beads in the containers are different from the beads from the hydrophobic interaction because on they have a different chemical called DEAE which makes them positively charged. GFP has both protons and electrons on it which is why it was not easily attracted t o the DEAE, which is why the GFP is put in a basic solution.Ata high pH, the amount of negatively charged hydroxide increases and these hydroxides are allured by the protons on the GFP. The protons are than neutralized, making GFP a negatively charged molecule. The isoelectric point of GFP is at a pH of 5. 3. Ata pH higher than 5. 3, it is negatively charged and when it is at a pH lower than 5. 3, it is positively charged. Once the column chromatography is filled with GFP and connected to a beaker of low salt which connected to a beaker of high salt, anion exchange occurs. As the salt concentration increases, the GFP slowly spreads down the column and eventually out f the column into test tubes.Between the HIC and the Ion exchange chromatography, the sample the fractions were pooled and put in a centricon causing the GFP concentration in the samples to increase. This occurred because the ultrafilter only allowed particles smaller than protein to go in to the pusher. The large protei ns stay in the centricon while buffer and salt solution goes into the plunger. The sample of GFP was also diluted 10 folds because the sample needs to be in a low salt solution to use anion exchange and after the GFP had gone through hydrophobic interaction, it was in a high salt solution.The anion exchange method creates a continuous salt gradient because as one drop of low salt solution goes into the column chromatography, causing GFP to spread down the container. The follow procedure was the three phase partitioning purification. T-butanol and 1. 6 molar ammonium sulfate were essential for this procedure. T-butanol has a low density causing in to stay above the GFP solution. In addition it has an attraction for water and other hydrophobic substances causing 5 mL of water to be drawn out of the GFP sample and precipitated substances to float between the t-butanol and the GFP sample.Fresh t-butanol is necessary after removing the old t-butanol with the contaminants because at that point, the salt concentration had increased since water was drawn out. was aDle to De preclpltatea Decause 0T tne nlgn salt concentration. The final procedure for purifying GFP was using the HPLC which separated substances by size. The beads used in the HPLC column are miniscule and porous. The pours on the beads give substances of the same size more opportunities to leave the HPLC at the same time. Since the beads are so small, high pressure is needed to push the GFP sample through the beads.Naturally, smaller substances will exit the HPLC first while larger materials will exist last. In all scientific experiments room for error is unavoidable. During the HIC, IEX, three phase partitioning, and the HPLC, amounts of GFP were lost due to the GFP sticking to a container, a pipette, and even spills. During the HIC some of the GFP was lost due the overflowing the test tubes with liquid exiting the column. During the HPLC some GFP was lost because not all GFP dripping out of the HPLC wen t in to the microphage. Other errors include letting the column dry because the liquid was not dded to the beaker about the column.During the spectrophotometer runs, the blank was no inserted correctly causing the reading of the GFP to be incorrect. In addition, the order in which the GFP samples were suppose to be placed in the spectrophotometer was messed up. Judging from the overall purification table, table 1, the purification was quite successful. Originally, the ratio was only . 25, but by the end of all the purification procedures, it obtained a ratio 1. 06. A 1. 25 ratio is most desirable and through the purification, the ratio was nearly reached. The anion exchange, three phase artitioning, and the HPLC purification were the most impacting procedures.The anion exchange greatly increased the purity of the crude sample compared to the HIC purification. The three phase partitioning and HPLC purified the GFP even more. Some improvements to the protocols would be to start with t he anion exchange purification so that overall, the salt solution would go from a low salt concentration to a higher salt concentration. This also eliminates the need to dilute the solution. In addition, an automatic machine could be used to shift the test tubes that collect the iquid exiting the columns to prevent overflowing test tubes and the risk losing GFP.GFP is unique because of its florescent glow. This glow can be used as a marker or an indicator. If a glowing marker could be placed on infectious cells such as tumor cells or cancerous cells, it would revolutionize the treatment of these diseases because doctors will be able to track where the harmful cells are. In addition, if it is possible to trigger the florescence of GFP with UV light, it can eventually be used in light bulbs to produce light. GFP light bulbs would last for an incredibly long time ince they are very resistant to denaturing.In addition, in vehicles, GFP can be mixed in the motor oil, transmission oil, po wer steering oil, air conditioning oil, and other oils so that if a leak occurs in a car, it can easily be spotted by shinning UV light on the car. The purification of GFP can lead to endless new innovations in electrical engineering, automotive repair, and curing deadly diseases.

Standard vs. Tipping Bucket Rain Gauges

A rain gauge is a  weather instrument that gathers and measures the amount of liquid precipitation that falls from the sky. How a Tipping-Bucket Gauge Works A tipping bucket rain gauge has several components that allow it to accurately measure rainfall. As rain falls, it lands in the funnel of the tipping bucket rain gauge. The rain travels down the funnel and drips into one of two very carefully calibrated ‘buckets’ balanced on a pivot (like a see-saw). The top bucket is held in place by a magnet until it has filled to the calibrated amount (usually approximately 0.001 inches of rain). When the bucket has filled to this amount, the magnet will release its hold, causing the bucket to tip. The water then empties down a drainage hole and raises the other to sit underneath the funnel. When the bucket tips, it triggers a reed switch (or sensor), sending a message to the display or weather station. The display counts the number of times the switch is triggered. Because it knows how much rain is needed to fill the bucket, the display can calculate the rainfall. Rainfall is measured in inches; 1 of rain would fill a container with straight edges to a level of 1. Getting the Best Results From Your Rain Gauge To get the most accurate results from a tipping bucket rain gauge, you need to properly install the rain gauge. The rain gauge must be positioned on a flat surface – if the surface isn’t flat, the see-saw may tip before the bucket has filled to the calibrated level, or not tip at all. If the bucket doesnt tip at the calibrated level, the rainfall calculated will not be correct. Use a spirit level to determine whether a surface is flat, and then fix the gauge to the flat surface to ensure you are getting an accurate reading.The rain gauge must be positioned on a surface that does not vibrate – surfaces such as a porch or fence can move and vibrate. The tipping bucket is very sensitive and any vibrations could cause the gauge to tip even if it is not raining.The instrument must not be positioned near trees – being positioned near trees could allow leaves or pollen to fall inside the funnel and block it, causing an inaccurate reading.It must not be positioned in a sheltered area – being positioned in a sheltered location (such us beside your house or a fence) co uld significantly increase or decrease the amount of rain depending on the wind direction, and cause an inaccurate reading. The gauge should be positioned at least twice as far away from the object as the object’s height (e.g. if the fence is 6 feet high, the gauge should be positioned at least 12 feet away).Your weather equipment must not be located near any magnetic, steel, or iron objects – magnetic, steel, or iron objects can affect the amount of time the magnet will hold the bucket or whether it will hold it all, causing an inaccurate reading. Will a Rain Gauge Measure Snow? If it snows where you live, most rain gauges will not be able to measure the snow fall; snow will block the opening of the collection funnel. However, special snow gauges are available to measure this. Following these recommendations should ensure your get an accurate result from your tipping bucket rain gauge.