Measurement of Anti-proliferative Activity Experiment

Measurement of Anti-proliferative Activity Experiment Human cancer cell lines A549 (Lung carcinoma), MCF-7 (Breast adenocarcinoma), DU 145 (Prostate carcinoma), DLD-1 (Colorectal adenocarcinoma), FaDu (squamous cell carcinoma of pharynx) were obtained from American Type Culture Collection (ATCC), USA. These cells were cultured in DMEM supplemented with 10% FBS and antibiotic combinations in 5% CO2 humidified atmosphere at 37 0C. A colorimetric sulforhodamine B (SRB) assay was used for the measurement of anti-proliferative activity as described before (Adaramoye et al., 2011; Fricker and Buckley, 1995; Keepers et al., 1991; Skehan et al., 1990). It is the second major technique for testing and is the more preferred. This basically depends on the incur of the negatively charged pink amino xanthine dye, sulphorhodamine B (SRB) through basic amino acids in the cells. The released dye will give a more intense colour and more absorbance, when the number of cells and amount of dye is taken up is greater, after fixing, when the cells are lysed, (Skehan et al., 1990). The SRB assay is sensitive, simple, reproducible and more rapid than the formazan-based assays and gives better linearity, a good signal-to-noise ratio and has a stable end-point that does not require a time-sensitive measurement, as do the MTT or XTT assays (Fricker and Buckley, 1995; Keepers et al., 1991). Ten thousand cells were seeded to each well of 96-well plate, grown overnight and exposed to test samples at 100  µg/ml concentration for 48 h. Cells were then fixed with ice-cold tri-chloro acetic acid (50% w/v, 50 µl/well), stained with SRB (0.4% w/v in 1% acetic acid, 50 µl/well), washed and air dried. Bound dye was dissolved in 150 ÃŽ ¼L of 10mM Tris base and plates were read at 510 nm absorbance (Epoch Microplate Reader, Biotek, USA). Anti-proliferative activity of test samples was calculated as: % inhibition in cell growth = [100-(Absorbance of compound treated cells/ Absorbance of untreated cells)] x100. Principal component analysis PCA was carried out based on the contents of eighteen bioactive compounds in fruits and leaves of five Cassia species, using STATISTICA 7.0 software. When the contents of investigated compounds were below the quantitation limit or not detected in the samples, the values of such elements were considered to be zero. Results and discussion Optimization of chromatographic and MS/MS conditions Complete separation of proximate analytes is certainly not required for MS/MS detection. In this study, chrysophanic acid and emodin are having same product ion, while catechin and epicatechin are having same precursor and product ion. Therefore, mobile phase was optimized using different compositions of solvents and adjusting their gradient elution for separation of all the compounds. Acetonitrile possesses stronger elution ability in comparison to methanol, which shortens the elution time and thus selected for this method. On the basis of the polarity of anthraquinones, phenolics, flavonoids and terpenoids in the extracts of Cassia species samples, an Acquity UPLC BEH C18 (2.1 mm Ãâ€" 50 mm, 1.7 µm; Waters, Milford, MA) column was selected for their separation, which was more suitable for acidic mobile phase with smoother baseline in the separation as compared to other tested columns. Compared with acetic acid, formic acid was found more effective for ionization of compounds det ected in the negative ESI mode. Thus, different concentration strengths (0.05%, 0.1% and 0.2%) of formic acid were investigated, and finally 0.1% formic acid concentration was selected for analysis. Therefore, optimized gradient elution with 0.1% formic acid in water and acetonitrile at a flow rate of 0.4 mL/min with the column temperature of 30 °C resulted in separation of the 18 compounds in less than 8 min chromatographic run time. All the compound dependent MS parameters (precursor ion, product ion, declustering potential (DP) and collision energy (CE) were carefully optimized for each targeted compound in negative ESI mode, which was performed by flow injection analysis (FIA). The chemical structures of 18 components were characterized based on their retention behaviour and MS information such as quasimolecular ions [M-H], fragment ions [M-H-COO], [M-H-COO-CH3], [M- CO-H2O] compared to related standards and literatures (Pandey et al., 2014; Wei et al., 2013; Xia et al., 2011; Yu et al., 2009). MRM parameters: DP, EP, CE and CXP were optimized to achieve the most abundant, specific and stable MRM transition for each compound as shown in Table 1. MRM extracted ion chromatogram of analytes are shown in Fig. 1. Analytical Method Validation The proposed UPLC-MRM method for quantitative analysis was validated according to the guidelines of international conference on harmonization (ICH, Q2R1) by linearity, LOQs and LODs, precision, solution stability, and recovery. Linearity, LOD and LOQ The internal standard method was employed to calculate the contents of eighteen analytes in Cassia species. The stock solution was diluted with methanol to different working concentrations for the construction of calibration curves. The linearity of calibration was performed by the analytes-to-IS peak area ratios versus the nominal concentration and the calibration curves were constructed with a weight (1/x2) factor by least-squares linear regression. The applied calibration model for all curves was y = a x + b, where y = peak area ratio (analyte/IS), x = concentration of the analyte, a = slope of the curve and b = intercept. The LODs and LOQs were measured with S/N of 3 and 10, respectively as criteria. The results were listed in Table 1. All the calibration curves indicated good linearity with correlation coefficients (r2) from 0.9990 to 0.9999 within the test ranges. The LODs for each analyte varied from 0.02-1.34 ng/mL and LOQs from 0.06-3.88 ng/ml and were much lower than those obtained with previous HPLC methods (Chewchinda et al., 2012; Chewchinda et al., 2014; Chewchinda et al., 2013; Ni et al., 2009; Prakash et al., 2007). Precision, Stability and Recovery The intra-day and inter-day variations, for the determination of precision of the developed method, were evaluated by determining the eighteen analytes in six replicates on a single day and by duplicating the experiments over three successive days. The overall intra-day and inter-day precision were not more than 3.37 %. Stability of sample solutions stored at room temperature was evaluated by replicate injections at 0, 2, 4, 8, 12 and 24 h. The RSDs value of stability of the eighteen analytes ≠¤ 3.19 %. A recovery test was applied to evaluate the accuracy of this method. Three different concentration levels (high, middle and low) of the analytical standards were added into the samples. Three replicates were performed at each level. The percentage recoveries were calculated according to the following equation: (detected amount–original amount) Ãâ€" 100% / added amount. The analytical method developed had good accuracy with overall recovery in the range from 97.75-105.09 % (RSD ≠¤ 2.42 %) for all analytes (Table 1).

Burke-Litwin: Understanding Drivers for Change Essay

There are many reasons that change occurs in organisations. Building on the Burke-Litwin model of organisational change and performance, this article will help you identify different drivers of change and consider the implications for you as a change manager. The Model The Burke-Litwin model[1] shows the various drivers of change and ranks them in terms of importance. The model is expressed diagrammatically, with the most important factors featuring at the top. The lower layers become gradually less important. The model argues that all of the factors are integrated (to greater or lesser degrees). Therefore, a change in one will eventually affect all other factors. The Burke-Litwin model Burke-Litwin believe environmental factors to be the most important driver for change. Indeed, most change can be traced back to external drivers for change. Important elements of organisational success, such as mission and strategy, leadership and organisational culture, are often impacted by changes that originate outside the organisation. It is your job to understand these external changes and identify the implications for you and your team. Identifying and Dealing with Drivers for Change 1. External Environment This includes such factors as markets, legislation, competition and the economy. All of these will have consequences for organisations, and, as a change manager, it is vital that you  continually scan the environment for issues that will affect you and your team. For example, in the world of accountancy, International Accounting Standards and International Financial Reporting Standards will have a significant impact on the way companies manage their accounts and report their results. In the public sector, legislative changes across health, local government and other services have a direct impact on the work organisations are required to carry out. 2. Mission and Strategy An organisation’s mission articulates its reason for existing. It is the foundation upon which all activity should be built. The strategy then sets out, in broad terms, how the organisation will go about achieving its mission. Very often, the strategy will be developed in light of environmental change, and will have a significant impact on the work you do. As a change manager, you need to understand change in strategy and be able to communicate the implications to your staff. 3. Leadership This considers the attitudes and behaviour of senior colleagues and how these behaviours are perceived by the organisation as a whole. The way in which change is implemented and accepted through the organisation will be largely influenced by the top team. Does your team believe that senior colleagues are committed to change, or is it just another initiative that will disappear in six month’s time? 4. Organisation Culture Organisation culture can be described as â€Å"the way we do things around here†. It considers the beliefs, behaviours, values and conventions that prevail in an organisation. Culture change does not happen overnight. It evolves over time as a result of many other changes in the organisation. As a manager, you should keep in mind the desired state for the organisation, in terms of how you expect people to behave (and not to behave), and what your organisation values as important. You need to ensure that your behaviour fits with these expectations at all times, and that you ‘walk the walk’. 5. Structure Very often, changes in strategy can lead to changes in the way the organisation is structured. This can impact on relationships, responsibilities and ways of working. Your job is to assess the impact of the structural change and ensure your team understands why it is required, and what it means for them. 6. Work Unit Climate This considers employees’ perception of their immediate colleagues and working environment. Our immediate working environment is often what shapes our view of the organisation as a whole, and influences the extent to which we feel satisfied in our jobs. Changes to the immediate working environment need to be managed sensitively, as they are likely to invoke a range of emotional and political responses from staff. This is particularly the case where change involves moving location, a change in personnel, or a change in terms of conditions of service, such as working hours. 7. Task Requirements and Individual Skills/Abilities Change at a higher level in the organisation will often require changes in the work carried out and the skills available in the team. As the change manager you need to assess whether: all the right skills are in place; if they can be developed; or, if you need to bring them in from outside the team. 8. Individual Needs and Values Changes to team membership can mean a change in the team dynamic. In a perfect world, we would be able to recruit the exact fit for our teams, in terms of personal style, abilities and skills mix. However, in reality it is not always possible, and it is your job to identify any risks in this areas and mitigate them as best you can. 9. Employee Motivation Considers the significance of individual and organisational goals. Motivation is key to effective change. The real challenge is to maintain motivation throughout a change project, particularly when change is often not well-received by those affected. [1] Burke & Litwin, ‘A Causal Model of Organisation Performance and Change’, Journal of Management, Vol 18, No 3 (1992), pp 523–545.

How Can Alternative Sources of Energy Be Harnessed

How can alternative sources of energy be harnessed effectively? Written by Jill (Huimei ZHOU) Nowadays, there are more and more alternative sources of energy for people to use. But in order to harness these alternative sources of energy effectively, different countries take different actions based on their own natural resources. This essay will show that how to make full use of energy from different countries' viewpoints. Undenied, many years ago, a lot of countries just use the fossil fuels to supply the energy that the whole city needs, but now the sources of energy are more than before, such as solar energy and biomass.So in this case, many countries try them best to harness the renewable energy to reduce the dependence on the traditional energy. Over the last five years, many European countries have increased their reliance on wind farms and hydroelectric dams because of more expensive fossil fuels. For example, in Sweden, the city Kristiantad is looking into building satellite b iogas plants for outlying areas and expanding its network of underground biogas pipes to allow the construction of more filling stations. [1]Besides the European countries, the United States also makes some measures to control the renewable energy more effectively. According to the Environmental Protection Agency, the United Sates need to spend about 8000 US farms on installing biomass plants. [2] In order to cut down costs, a number of states and companies are considering new investments or programs. They hope to build plants in Calnifornia to turn organic waste from farms and gas from water treatment plants into biogas. [3] Because of less natural energy and more expensive fees, this way can help the whole state to meet requirements.Above on these countries' methods, we can clearly see that different countries have different environments and situations. So if a country want to harness alternative sources of energy, they should make more reasonable policies depending on their real conditions, which will be more useful. Reference: [1]Using waste, Swedish city cuts its fossil fuel use[1], Source Book, Paragraph 3. [2]Using waste, Swedish city cuts its fossil fuel use[1], Source Book, Paragraph 4. [3]Using waste, Swedish city cuts its fossil fuel use[1], Source Book, Paragraph 4.

Martin Luther and the Reformation - 2927 Words

Martin Luther and the Reformation A German Augustinian friar, Martin Luther launched the Protestant Reformation of the sixteenth century. Luther grew up the son of a miner, but he did not maintain that lifestyle for himself. He lived in a period that had a widespread desire for reformation of the Christian church and a yearning for salvation. Martin Luther was born at Eisleben in Saxony. Since his father was a miner, it was a great distress on him to send Martin to school and then to the University of Erfurt. There is where he earned his masters degree at the young age of twenty-one. (Erikson, 39) Although his father wished him to study law, Martin, after being terribly frightened in a thunderstorm, vowed to become a friar. In†¦show more content†¦Archbishop Albert needed someone to sell indulgences for him, so he hired Dominican friar John Tetzel. Tetzel made this into a business and even began to heavily advertise the sale of indulgences. He came up with catchy slo gans to lure people into buying them. He even came up with a chart price to persuade people to buy the best or most expensive indulgences. Luther did not agree with this because people no longer saw the need for repentance and felt they could buy away their sins. He was greatly troubled that people were buying into these advertising gimmicks. Since, at the time, the church did not have an official doctrine on indulgences, Luther decided this entitled him to discuss the subject critically. In doing so Luther wrote Archbishop Albert a letter on the subject and enclosed in Latin Ninety-five Theses on the Power of Indulgences. He argued indulgences made people believe repentance was not important, it downplayed the importance of charity in Christian life, and it competed with the preaching of the Gospel. (Boehmer, 198) Once Luther died, his disciple Philipp Melanchthon reported that the theses were also posted on the door of the church at Wittenberg Castle on October 31, 1517, al though not all modern scholars are completely convinced this ever took place. (Erikson, 142) By December of 1517, the theses had all been translated to German and were read throughout the empire.Show MoreRelatedMartin Luther And The Reformation1803 Words   |  8 PagesThesis Statement The purpose of my paper will reflect the life of Martin Luther during the Reformation period and the changes brought forth as a result of his teachings and writings. Luther’s findings not only transformed his own actions, but change the thought process of millions. The study of Martin Luther is the key that unlocks the background of the Protestant Reformation and it’s origins. Introduction Research on Martin Luther was chosen to demonstrate the importance of his works, and decisionsRead MoreThe Doctrines Of Martin Luther, Reformation And The Reformation740 Words   |  3 PagesLuther was ordained in 1508. In 1512, he became a lecturer at the university of Wittenberg. Significantly, Luther’s devotion to theology would lead him to explore the problems plaguing the Catholic Church. During his inquisitive years, Luther began to question the doctrines of the Catholic Church. Luther’s view of salvation, penance and righteousness began to change from the view he was instructed to believe in. Additionally, Luther’s extensive study led him to form his own ideas and formulate aRead MoreMartin Luther And The Reformation Of The Protestant Reformation Essay1699 Words   |  7 PagesMartin Luther became one of the most influential figures in Christian history. He began the protestant reformation in the 16th century. He also called into question some of the b asic tenets of Roman Catholicism and eventually his followers soon split from the Roman Catholic Church to become protestant. He was known to be a very controversial man not just for his writing of the ninety five thesis but for others and in his later years his feeling about the Jews. His teaching of the bible is what leadRead MoreMartin Luther And The Protestant Reformation1349 Words   |  6 Pagesto fix by amendment is Martin Luther. In the early 15th century, the Catholic Church was under siege from criticism based off of corruptions and Catholic wrongdoings; this time in history was known as the Protestant Reformation. The main catalyst in this revolution was none other than Martin Luther. Although Martin Luther sparked rebellion and a revolution, he , himself, was a reactionary reformer in how he wanted to restore the church into what it once was. Martin Luther, according to his ideasRead MoreMartin Luther And The Protestant Reformation1608 Words   |  7 Pagesfirst step o f the Protestant Reformation was carried out by a monk called Martin Luther to adjust the unfairness of the Catholic Church. He believed that the Christian faith was a simple religion, misrepresented on account of inadequate papal authority. Like the Roman Empire the Church had broadened its territorial domain and bureaucratic function. The moral authority of the church was corrupted because of its tainted clerical practices. In his Ninety-Five Theses, Martin mainly targeted the doctrineRead MoreMartin Luther And The Protestant Reformation1326 Words   |  6 PagesThus the Religious Reformation period was the most important time because new religions were formed and the RCC had all the power. Indeed supporters of the Reformation Religions believe that this time had the biggest impact because during this time new religions were created and millions changed faith. In his text â€Å"Martin Luther† the author argues that â€Å"Martin Luther as a Christian theologist and Augustinian monk whose teachings inspired the Protestant Reformation†. Martin Luther was the spark forRead MoreMartin Luther And The Protestant Reformation1879 Words   |  8 PagesMartin Luther was not only one of the most significant figures in the Protestant Reformation, but he also developed his own denomination of followers. While many say that Luther was breaking away from the Catholic Church to establish a rival church, yet Luther challenged the authority of the church quietly. His arguments did not focus the attention on himself, but wholly on God. Martin Luther is considered the initiator of the Protestant Reformation as he realized the corruption in the Catholic ChurchRead MoreMartin Luther And The Protestant Reformation Essay2089 Words   |  9 Pages1517, as the initial rise of early reform movements. It was the day Martin Luther had encrypted his floating thoughts of early reformation onto paper, with the help of the printing press. Many others before Martin had speculation of somewhat of a reform, but didn’t have the courage to go up against the authoritative Church. Martin and his actions had officially sparked the revolution, and the start of European Reformation. But Martins actions would end up transforming the world indefinitely. SuccessorsRead MoreThe Art Of Reformation By Martin Luther985 Words   |  4 PagesAnalytical Paper: The Art of Reformation The year 1517 is claimed by Christianity to be the original spark of a reformation that changed the face of the most prevalent religion in the western world, beginning in Germany and soon enveloping the known world in an upheaval of beliefs and values. However, due to the fact that Protestant faiths became their own religious, social and political powers, the correct term for this movement would not be ‘reformation’ but in fact ‘revolution’. The so-calledRead MoreMartin Luther And The Protestant Reformation1541 Words   |  7 PagesSochor Professor Eric Breault Religion 374 21 April 2017 Martin Luther and the Protestant Reformation Martin Luther was a German professor of Theology during the 13th and 14th century who was a key figure of the Protestant Reformation. In this paper, I will discuss the impact of Martin Luther’s actions from a standpoint of the Catholic Church and its reaction to Martin Luther’s work. To discuss this, I will use some of the works of Martin Luther himself along with sources from authors Paul A. Bishop