Anti-cancer Drug Targeting AFR Receptor

Anti-cancer Drug Targeting AFR Receptor Results 1a 1b Figure 1 Cell viability data based on the use of the XTT assay Two different lung tumor cell lines were tested: one expressing the mutated AFR receptor (blue line) and one that expresses wild-type AFR (orange line). Figure 1a represented AFR lung cancer cell line with Drug X and figure1b represented AFR lung cancer cell line with drug Y. Cells were seeded in wells of a 96 well plate at 3 x 10*4 cells/well before treatment with the drugs at the indicated concentrations for 72 hours (triplicate samples at each concentration). XTT assay reagent was then added to the wells and the absorbance measured at 450 nm. Control wells (no cells) contained medium but no cells. The Cell Proliferation XTT assay is a colorimetric assay for the nonradioactive analysis of cellular proliferation, viability, quantification of cytotoxic and cytostatic compounds like anticancer drugs and pharmaceutical compounds, evaluation of growth-inhibitory antibodies and physiological mediators that able to inhibit cell growth (Sigma-Aldrich, 2016). Sample is adherent or suspension which cultured in 96-well microplates. A fictional receptor (AFR) was targeted by anti-cancer drug to inhibit the mutated AFR function, causing an anti-proliferative effect and even cell death. XTT ((2,3-bis-(2-methoxy-4-nitro-5-sulfophenyl)-2H-tetrazolium-5-carboxanilide)) assay was used to measure cell viability in the two different lung tumor cell lines, one expressing the mutated AFR receptor (AFR mut) and one that expresses wild-type AFR (AFR wt). Drug Y was observed to hinder cancer cell growth dose dependently (Figure 1). The amount of observed absorbance increases with cell number and incuba tion time for 72hours when a maximum absorbance level is reached. The plot of the XTT assay data should create a curve with a linear part. This area exhibits the highest sensitivity to changes produced by the experimental parameters. Establishing the XTT assay data for treatment of AFR mutated and AFR wild-type lung cancer cell line with drug X from 0nm to 1600nm (Figure 1a), It was illustrated that the cell viability of mutated AFR increased considerably from 1.1 to 1.3 between 0nm and 800nm and then became steady at approximately 1.4 from 800nm to 1600nm. Compared with the wild type of AFR (Figure1a), the trend of the line was similar to the mutated one but the cell viability was first rised from 0.9 to 1.2 between 0nm and 800nm and finally exhibited a loss of linearity when greater than 800nm. The lung cancer cell line with drug X represented a cell proliferation assay because the absorbance values of the mutated one were greater than control conditions, representing an increase in cell proliferation and viability. On the other hand, investigating the XTT assay data for treatment of AFR mutated and AFR wild-type lung cancer cell line with drug Y from 0nm to 1600nm, It was indicated that the cell viability of m utated AFR started off at about 1.1 from 0nm to 50nm and then occurred a dramatic drop from 1.1 to 0.1 between 50nm to 400nm and finally lied on 0.03 at 1600nm (Figure1b). Compared with the wild type of AFR (Figure1b), the line was also decreasing but showed a completely different trend. The cell viability decreased from 0.89 to 0.88 between 0nm and 800nm stably and then exhibited a significant drop when greater than 800nm and finally lied on 0.8 of 1600nm. The lung cancer cell line with drug Y represented a cell apoptosis assay as the absorbance values were lower than control conditions, showing a decrease in cell proliferation and resulted from cellular necrosis or apoptosis. From the data analysis, drug Y was successfully tested to inhibit the function of mutated AFR which have an anti-proliferative effect to cause cell death and drug Y could be put forward for further testing. Figure 2 Flow cytometry apoptosis data: Annexin V -FITC (x axis) and propidium iodide (Y axis)   Ã‚   The proapoptotic effect of drug X and Y. The AFRmut cell line was treated with each drug at a concentration of 800 nM for 24 hours. The cells were harvested, resuspended in buffer and then treated with Annexin V-FITC and propidium iodide. Cytograms of annexin-V-FITC binding against PI uptake show three populations: (i) viable cells (low FITC and low PI signal) in gate Lower Left; (ii) early apoptotic cells (high FITC and low PI signal) in gate Lower Right and (iii) cells that lost membrane integrity which results in late apoptosis (high FITC and high PI signal) in gate Upper Right. The percentage of cell death increases in the tumor cancer cell line of drug Y, as demonstrated by incorporation of Annexin V. The inhibition of cell proliferation caused by AFRmut was related to the apoptosis induction. Apoptosis is programmed as cell death which is important in development, in normal function of certain tissues and in the response to the damage. Apoptosis involves events which results in the destruction of cell. Annexin V -FITC is a protein that binds to phosphotidylserine which can be labelled and used to detect apoptotic cells. The flow cytometry apoptosis data was plotted on a dot plot and the lower left was represented the viable cells while the upper right and the lower right were represented the dead cells and the apoptotic cells respectively. From the apoptosis data (Figure2), it was observed that the untreated condition showed 77% viable cells and 21% of apoptotic effect. When AFRmut was treated with drug X, the viable cells were increased by 19% and the apoptotic effect was largely decreased to 3%. In contrast, the incorporation with drug Y lead to considerable anti-proliferative effect where 58% of apoptotic cells were observed and viable cells only remained as 40%. Related to the induction of programmed cell death, the majority of cells from the two tumor cell lines were alive upon carrier treatment, the use of AFRmut lead to the entrance of these cells into apoptosis with incorporation of Annexin V. The treatment of drug Y showed a significant apoptotic effect than the drug X, which has a potential to inhibit the function of mutated AFR and results in anti-proliferative effect. Figure 3 Flow cytometry cell cycle data: x-axis shows propidium iodide fluorescence and y-axis shows the number of cells The AFRmut cell line was treated with drug X and Y at a concentration of 800 nM for 24 hours and then fixed with ethanol and stained with propidium iodide before analysis in a flow cytometer. Treating with drug Y showed no cells in G2/M phase. The inhibition of cell proliferation induced by AFRmut was correlated with cell cycle which was evaluated with propidium iodide by flow cytometry. Cell cycle demonstrated the cell progression through a division cycle which result in cell growth and separation into two daughter cells. Live cell observation of cellular DNA and distribution of cell cycle are valuable to regulate apoptosis, and also the tumor behavior and suppressor gene systems. Cells would be distributed into three phases of cell cycle: G0 /G1 phase (2N), S phase (DNA synthesis with various DNA), and G2 /M phase (4N) to identify apoptotic cells with partial DNA content desirably   (Krishnakumar R, Kraus W ,2010). Propidium iodide fluorescence was needed in these actions to bind DNA. From the flow cytometry results by histogram (figure 3), the untreated condition showed 42% of cells in G0/G1 phase while 31% in S phase and 27% in G2/M phase. When the AFRmut was treated with drug X, the cell number in G1 phase dropped by 7% while there was increase of 6% in S Phase. DNA replication occurs more effectively during S phase. Tumor cells with a higher proportion of cells in S phase as tumors were growed faster and more aggressive in that phase so drug X induced proliferation of mutated AFR. However, the addition of drug Y caused effective anti-proliferation effect by increasing the cell numbers in G1 phase from 42% to 86% since the cell has left the cycle and has stopped dividing. There was increase in the cell numbers and grow in size. The decrease from 31% to 14% for S phase indicated the ineffective DNA replication. Moreover, there was no cells observed in G2/M phase when treated with drug Y which represented the increasing numbers of apoptotic cells and the G2/M pha se arrest, which failed the mitosis progression. Discussion Drug Y can be an effective anticancer drug which targets mutated AFR and inhibit its function to lead an anti-proliferative effect. In the present study, two lung tumor cell lines, mutated AFR receptor (AFRmut) and wild-type AFR (AFRwt), Drug Y was found that it could inhibit the growth of the AFR in vitro. The IC50 values of AFRmut and AFRwt were with the same range of 450nm. Additionally, the cell cycle revealed that Drug Y arrested mutated AFR at S phase and G2/M phase. Furthermore, the data demonstrated that Drug Y induced apoptosis in the mechanism assessed by 3 different methods: cell viability assay, apoptosis assay and the cell cycle of flow cytometry. Taken together, these results suggest that the inhibitory effect of Drug Y on cell proliferation in mutated AFR is mediated through induction of cell cycle arrest and apoptotic cell death. The limitation is the flow cytometry are only authorize the relative abundances of cell cycle phases compared to another (Dick FA, Dyson NJ, 2002). These actions can be examined by evaluating a coordinate sample of cells with mitotic inhibitor like nocodazole or G1/S inhibitor like aphidicolin. As these drugs establish a prevalent arrest in M-phase or early S-phase accordingly, moderately proliferating cells will gather at the drug inferred arrest location. For instance, cells arrested in G1 BY pRB expression will stay in G1 phase regardless of nocodazole treatment even regulating cells will accumulate in M-phase[13] Overall, the data demonstrated drug Y is able to develop an anti-cancer drug that targets a receptor called AFR (A Fictional Receptor) to block its activity. Drug Y successfully inhibits the function of mutated AFR by evaluating the cell viability assay, apoptosis assay and the cell cycle of flow cytometry. The assays showed an anti-proliferative effect and even cause cell death, which explains its significant antitumor activity in the various experimental investigations that have assessed. Thus, further studies about the way of administration, dose limiting toxicities and recommended doses are needed for the preclinical development of drug Y to evaluate the potential of the compound as an anticancer drug. References Dick FA, Dyson NJ, 2002. Three regions of the pRB pocket domain affect its inactivation by human papillomavirus E7 proteins. J. Virol.2;76: 6224-6224. Krishnakumar R, Kraus W, 2010. The PARP side of the nucleus: molecular actions, physiological outcomes, and clinical targets. Mol Cell. 39:8-24 Matthew J., et al, 2012. Analysis of Cell Cycle Position in Mammalian Cells  (59): 3491. Sigma-aldrich, 2016. Cell Proliferation Kit II (XTT) [online] Available at: http://www.sigmaaldrich.com/catalog/product/roche/11465015001?lang=enregion=GB Wang, G., et al, 2005. Antitumor effect of ÃŽÂ ²-elemene in non-small-cell lung cancer cells is mediated via induction of cell cycle arrest and apoptotic cell death. Cellular and Molecular Life Sciences CMLS 62.7-8: 881-893.

Effects of electronic gadgets to students studying habits Essay

Abstract Almost all of the students especially high school students are seen of using different kinds of electronic gadgets everyday. Like cellphone, iPad, psp, etc. With that case, students who are addicted in using electronic gadgets affects their studies. As what I have observed, using of electronic gadgets gives bad effects to almost all of the students. The purpose of doing this study is to *Explain the disadvantages of using electronic gadgets among us students *Determine the positive and negative effects of Electronic gadget to students *Show the consequences of excessive use of Electronic gadgets to students *Give the students an advice on how to control their use of electronic gadget. To accomplish this study, I researched for some information that can help me to prove the content of my study. And to be able to solve this problem, I researched for some solutions that can help me to solve this kind of situation, which may also help me to show the goal of my study to my co-students who are addicted in using electronic gadgets. In that way, I can help to prevent the addiction of every student from electronic gadgets. Effects of electronic gadgets 3 EFFECTS OF ELECTRONIC GADGETS TO STUDENTS STUDYING HABITS â€Å"Modern technologies like television and computers provide identifiable educational advantages, such as greater access to information and more compelling presentations of that information. Over-use of technology, though, especially such gadgets as cell phones, iPods and video games.

Army Strategic Planning Guidance - Free Essay Example

Sample details Pages: 4 Words: 1061 Downloads: 5 Date added: 2019/10/10 Did you like this example? Introduction The chief purpose of specialized Explosive Ordinance Disposal (EOD) force is executed missions within the United States and all over the world in combating military weapons that can cause mass destruction of property as well as the lives of people. The force is committed to protect and maintain peace in the United States and globally at a large by limiting the supply of weapons that can endanger the lives of the public in case the weapons land on the hands of the wrong people. Based on the available Army strategic priorities; I would highly recommend for an Army that has the ability to change and cope with the challenging situations they are facing in their military duties. This is the only priority among the five strategic priorities that appreciates change is inevitable and for that reason, the future holds a lot of uncertainties and therefore the Army should strive to be ready to operate efficiently under circumstances of uncertainties. The defense forces should be prudent in predicting the new emerging threats the Nation is likely to face and for that reason formulate mitigation measures to curb the attack before it occurs. The world has become a complex environment due to increased production of weapons that endanger the lives of people and animals[footnoteRef:2]. That means developing an Army that is flexible enough to respond effectively to the emerging threats to security matters should be the top priority in order to achieve peace and harmony in t he United States and therefore the other strategic goals will thrive well in presence of peace and harmony. Dynamic army force will enable the leaders to make the critical judgment under the unfamiliar conditions which will pave way for other strategic priorities since any unseen obstacle will be easily spotted and effectively counteracted before it results in a significant loss. Don’t waste time! Our writers will create an original "Army Strategic Planning Guidance" essay for you Create order The Army Strategic Planning is undertaking a number of measures to enhance the capability of dynamic army force in the advent of changing war tactics. The following are the number of measures that the Army is undertaking in an attempt to successfully adopts this strategy. First, reforms are being made in Army profession in the 21st century. This is evident through, retaining the gains of the previous decades and also eliminating the harmful leadership and the negative effects of long experienced war Army leaders who depicted professional leadership and skills are been retained in order to disseminate the same values and skills to the current Army in order to facilitate accurate and critical judgment making the ability to under challenging times of uncertainties. Secondly, a lot of learning programmes are been established to educate the leaders for an uncertain future. The endeavor is undertaken in conjunction with other multinational and inter-agency groups. Lastly, leaders are being developed who are competent enough in Cyberspace. The leaders are expected to understand and be swift enough to adapt to constantly volatile capabilities and risks. It is highly stressed that while recruiting, educating training and retaining cyberspace personnel in the entire Army, it will be very crucial to create a strong Army where the force will have a great deep of expertise in cyberspace as well as cyber-electromagnetic activities. Developing persevering soldiers, dynamic leaders, and united army force guided by the code of ethics outlined under the army force code of regulations that will ensure successful accomplishment of different kinds of missions under unfamiliar conditions and changing environments. This is the required capability that needs to be given the highest priority for the Army to operate efficiently and in a complex world. A lot of efforts are needed to support the mentioned strategy by the Army because the strategy demands a collaborative effort across the established Armed forces. Secondly, future Army force has to formulate training programmes that will equip the Army personnel with the right skills and knowledge that will enable the Army to remain resilient under challenging times and endeavor to accomplish the set mission. In addition to that, efforts should be put in place in order to enlighten how forces and those that hold Army leadership positions combine the military resources across war-fighting activities an attempt to achieve the set mission. There are several improved capabilities that the Army can develop as part addressing the capability that can be used by the Army. The following is one of the improved capabilities that the Army should develop in an attempt of curbing the hurdles confronting the discharge of their duties. The institution should consider optimizing human performance as well as ensuring the each soldier is assigned to the right assignment at the correct time consequently maximizing the possibility of accomplishing the mission as per the planned schedule. Institutional as well as operational Army should endeavor to develop creative leaders and increase their performance which will enhance the success of the missions that the Army will undertake. Creative leaders can be developed through subjecting trainee on repetitive training programmes joined with self-study, thorough and careful education all geared at developing innovative leaders that will be resilient and effective in challenging times. Leaders ar e known to impart trust among their fellow leaders and soldiers. Additionally, they develop a spirit of togetherness that fuels the urge of taking the initiative that conforms to the goal of targeting to achieve the mission despite the obstacles that might hinder the success of accomplishing the mission[footnoteRef:3]. Leaders along with Soldiers are accountable to each other and the code of ethics set that demands soldiers to remain persevering and maintain moral character as they perform Army duties. Finally, the implications of potential non-material solution approach recommendations aim at evaluating how it analysis the military combats with emphasize an action warfare and joined air ground activities that aim at evaluating whether there is better an optimal way that can solve a current capability gap. The first implication is that the training analysis evaluates how the Army forces are strategized to fight, groups, air wings task forces and other army groups that are playing a crucial role in contributing to the accomplishment of the mission planned. Secondly, the material analysis explores the entire required equipment and systems that are vital to the forces to facilitate effective execution of the set mission and if new systems are required to meet the existing gap. Next, the leadership and education is the key component that needs to address.

What Is Narrative Poetry Definition and Examples

Narrative poetry tells stories through verse. Like a novel or a short story, a narrative poem has plot, characters, and setting. Using a range of poetic techniques such as rhyme and meter, narrative poetry presents a series of events, often including action and dialogue. In most cases, narrative poems have only one speaker—the narrator—who relates the entire story from beginning to end. For example, Edgar Allan Poes The Raven is narrated by a grieving man who, over the course of 18 stanzas, describes his mysterious confrontation with a raven and his descent into despair. Key Takeaways: Narrative Poetry Narrative poetry presents a series of events through action and dialogue.Most narrative poems feature a single speaker: the narrator.Traditional forms of narrative poetry include epics, ballads, and Arthurian romances. Origins of Narrative Poetry The earliest poetry was not written but spoken, recited, chanted, or sung. Poetic devices like rhythm, rhyme, and repetition made stories easier to memorize so they could be transported long distances and handed down through generations. Narrative poetry evolved from this oral tradition. In nearly every part of the world, narrative poetry established a foundation for other literary forms. For example, among the highest achievements of ancient Greece are The Iliad and  The Odyssey, which have inspired artists and writers for more than 2,000 years. Narrative poetry became an enduring literary tradition throughout the Western world. Composed in Old French, Chansons de geste (songs of deeds) stimulated literary activity in medieval Europe. The German saga now known as the Nibelungenlied lives on in Richard Wagners lavish opera series, The Ring of the Nibelung (Der Ring des Nibelungen). The Anglo Saxon narrative Beowulf has inspired modern-day books, movies, operas, and even computer games. In the East, India produced two monumental Sanskrit narratives. The Mahabharata is the worlds longest poem with over 100,000 couplets. The timeless Ramayana spreads Indian culture and ideas across Asia, influencing literature, performance, and architecture. Identifying Narrative Poetry Narrative is one of three major categories of poetry (the other two being dramatic and lyric), and each type of poetry has distinct characteristics and functions. While lyric poems emphasize self-expression, narrative poems emphasize plot. Dramatic poetry, like Shakespeares blank verse plays, is an extended stage production, usually with many different speakers. However, the distinction between genres may blur as poets weave lyrical language into narrative poems. Similarly, a narrative poem might resemble dramatic poetry when the poet incorporates more than one narrator. Therefore, the defining feature of narrative poetry is the narrative arc. From the epic tales of ancient Greece to 21st-century verse novels, the narrator moves through a chronology of events from challenge and conflict to a final resolution. Types of Narrative Poems Ancient and medieval narrative poems were most commonly epics. Written in a grandiose style, these epic narrative poems retold legends of virtuous heroes and powerful gods. Other traditional forms include Arthurian romances about knights and chivalry and ballads about love, heartbreak, and dramatic events. However, narrative poetry is an ever-evolving art, and there are countless other ways to tell stories through verse. The following examples illustrate several different approaches to narrative poetry. Example #1: Henry Wadsworth Longfellow, "The Song of Hiawatha" On  the  Mountains  of  the  Prairie,On  the  great  Red  Pipe-stone  Quarry,Gitche  Manito,  the  mighty,He  the  Master  of  Life,  descending,On  the  red  crags  of  the  quarryStood  erect,  and  called  the  nations,Called  the  tribes  of  men  together. The Song of Hiawatha by American poet Henry Wadsworth Longfellow  (1807–1882) narrates Native American legends in metrical verse that mimics the Finnish national epic, The Kalevala. In turn, The Kalevala echoes early narratives such as The Iliad, Beowulf, and the Nibelungenlied.   Longfellows long poem has all the elements of classical epic poetry: a noble hero, a doomed love, gods, magic, and folklore. Despite its sentimentality and cultural stereotypes, The Song of Hiawatha suggests the haunting rhythms of Native American chants and establishes a uniquely American mythology. Example #2: Alfred, Lord Tennyson, "Idylls of the King" â€Å"I fain would follow love, if that could be;I needs must follow death, who calls for me;Call and I follow, I follow! let me die.† An idyll is a narrative form that originated in ancient Greece, but this idyll is an Arthurian romance based on British legends. In a series of twelve blank verse poems, Alfred, Lord Tennyson (1809–1892) tells the story of King Arthur, his knights, and his tragic love for Guinevere. The book-length work is drawn from medieval writings by Sir Thomas Malory. By writing about chivalry and courtly love, Tennyson allegorized behaviors and attitudes he saw in his own Victorian society. Idylls of the King elevates narrative poetry from story-telling to social commentary. Example #3:  Edna St. Vincent Millay, "The Ballad of the Harp-Weaver" â€Å"Son,† said my mother,  When I was knee-high,  Ã¢â‚¬Å"You’ve need of clothes to cover you,  And not a rag have I.  Ã¢â‚¬Å"There’s nothing in the house  To make a boy breeches,Nor shears to cut a cloth with  Nor thread to take stitches. The Ballad of the Harp-Weaver tells the story of a mothers unconditional love. By the end of the poem, she dies weaving her child magical clothes from her harp. The mothers dialog is quoted by her son, who placidly accepts her sacrifice. American poet Edna St. Vincent Millay (1892–1950) cast the story as a ballad, a form that evolved from traditional folk music. The iambic meter and a predictable rhyme scheme of the poem create a sing-song rhythm that suggests child-like innocence. Famously recited by country musician Johnny Cash, The Ballad of the Harp-Weaver is both sentimental and disturbing. The narrative poem can be understood as a simple story about poverty or a complex commentary on the sacrifices women make to clothe men in the garbs of royalty. In 1923, Edna St. Vincent Millay won a Pulitzer Prize for her poetry collection of the same title. Story song ballads became an important part of the American folk song tradition of the 1960s. Popular examples include Bob Dylans Ballad of a Thin Man and Pete Seegers Waist Deep in the Big Muddy. Example #4: Anne Carson, "Autobiography of Red"   Ã¢â‚¬Å"†¦Small, red, and upright he waited,gripping his new bookbag tightin one hand and touching a lucky penny inside his coat pocket with the other,while the first snows of winterfloated down on his eyelashes and covered the branches around him and silencedall trace of the world.† Canadian poet and translator Anne Carson (b. 1950) loosely based Autobiography of Red on an ancient Greek myth about a heros battle with a red-winged monster. Writing in free verse, Carson recreated the monster as a moody boy who battles modern-day problems related to love and sexual identity. Carsons book-length work belongs to a genre-jumping category known as the verse novel. It shifts between description and dialogue and from poetry to prose as the story moves through layers of meaning. Unlike long verse narratives from antiquity, novels in verse do not adhere to established forms. Russian author Alexander Pushkin (1799–1837) used a complex rhyme scheme and an unconventional meter for his verse novel, Eugene Onegin, and English poet Elizabeth Barrett Browning (1806–1861) composed Aurora Leigh in blank verse. Also writing in blank verse, Robert Browning (1812–1889) composed his novel-length The Ring and the Book from a series of monologues spoken by different narrators. Vivid language and simple stories have made book-length narrative poetry a popular trend in young adult publishing. Jacqueline Woodsons National Book Award-winning Brown Girl Dreaming describes her childhood as an African American growing up in the American South. Other best selling verse novels include The Crossover by Kwame Alexander and the Crank trilogy by Ellen Hopkins. Sources Addison, Catherine. The Verse Novel as Genre: Contradiction or Hybrid? Style. Vol. 43, No. 4 Winter 2009, pp. 539-562. https://www.jstor.org/stable/10.5325/style.43.4.539Carson, Anne. Autobiography of Red.  Random House, Vintage Contemporaries. March 2013.Clark, Kevin. Time, Story, and Lyric in Contemporary Poetry. The Georgia Review. 5 March 2014. https://thegeorgiareview.com/spring-2014/time-story-and-lyric-in-contemporary-poetry-on-the-contemporary-narrative-poem-critical-crosscurrents-edited-by-steven-p-schneider-patricia-smiths-shoulda-been-jimi-savannah-robert-wr/Longfellow, Henry W. The Song of Hiawatha. Maine Historical Society. http://www.hwlongfellow.org/poems_poem.php?pid62Tennyson, Alfred, Lord. Idylls of the King. The Camelot Project.  University of Rochester. https://d.lib.rochester.edu/camelot/publication/idylls-of-the-king-1859-1885