Essays of Pochury Festival

STORY BEHIND SEPTEMBER 6 BLACK DAY| | On August 14, 1960, the Naga Army assaulted Assam Rifle out-post at Thuda (Phor town) in Pochury region. The assault was embraced by the Eastern Command’s first Brigade under the order of Major General Zuheto, alongside the then fourth contingent of Pochury Region under the order of Lt. Colonel Thorpa. The storm was at its stature during the time and every single significant stream like Tizu, Lanye and Thethsii were in full spate. The assault was propelled in the wake of devastating all the six extensions on all the streams. This was done to prevent fortification from arriving at the assaulted armed force post.As the assault proceeded into the thirteenth day, ammo on the two sides were running low and on a few events the Indian Air Force plane attempted to drop alleviation material and ammo yet were forestalled by the Naga Army. Simultaneously the Indian Air Force fly warriors barraged the attacker’s positions. An Indian vehicle pla ne (Dakota) attempting to drop help materials and ammo to the attacked post was shot somewhere around the Naga Army on the fourteenth day of the attack and crash arrived at Zathsii, a paddy field of Meluri town. The Naga Army caught all the 9 (nine) aviators including Flt.Lt. A. S. Singh. This prompted an overwhelming armed force activity in Pochury territory by the Indian Army, who was set for search and salvage the caught pilots, none of whom were ever tormented yet were later liberated through the Red Cross. During the time spent the military tasks to protect the pilots numerous towns were burned to the ground and untold outrages and torments were dispensed upon the residents. On September 1, 1960, 6 (six) residents from Phor town were tormented to death. Their names are Lt. Turachu, Village Chief, Lt. Yutsuchu, Pastor, Lt. Chupuchu DB, Lt. Yituchu GB, Lt. Turuchu GB, Lt. Mughazu GB.Again on September 3, 1960 another 3 (three) townspeople from Yisi town were pounded the life out of in particular Lt. Mazu GB, Lt. Throchu, Lt. Mazu RP. Two residents Lt. Yichuhu and Lt. Nyupuchu. from Mokie town were likewise pounded the life out of. In Laruri town, Lt. Lingsang was covered alive after extreme beating. Lt. Nyukhrusuh and Lt. Rhorupa of Meluri town were beaten harshly and after which, their heads were hacked off. Two towns, to be specific Tsikuzo and Kuluopfu, were relinquished because of torments and embarrassments dispensed by the Indian armed force. On sixth September 1960, the Punjab Regiment posted at Kangjang town came to Matikhrii town around 10 am.The whole town was surrounded in three rings and all the residents were requested to accumulate in one spot. Men people were isolated from ladies and kids. All the men were made to continue bouncing and do sit-ups, for over 5 hours in the burning sun, bare. Any indications of sluggishness were met with kicks and hits with rifle handles. At that point not long before dusk, Indian armed force not happy with the discipline dispensed to the townspeople, gathered them together inside the Village chief’s house and had to sit heads down like a sheep being lead to its butcher. Lt.Thah, the then Village head, realizing what was coming up for them fearlessly elected to forfeit. He stood valiantly for the Naga cause even to his final gasp and said â€Å"It’s a man’s pride. No acquiescence, no trade off for our inheritance. This penance is to ensure our opportunity. I will readily set out my life for the Naga group of people yet to come. † Then an Indian armed force jawan, holding an unpolished dao(hatchet) slashed off the head of Lt. Pogholo who was first in the line. Seeing the ruthlessness and repulsiveness before their eyes and realizing that every one of them would have been slaughtered, one of the townspeople figured out how to get away from the execution forcefully.Then in a steady progression heads moved down isolated from the bodies, and in the occasion a sum of nine lives were lost. Their names are Lt. Thah, Lt. Pogholo, Lt. Mezitso, Lt. Pongoi, Lt. Eyetshu, Lt. Zasituo, Lt. Thitu, Lt. Kekhwezu, Lt. Kezukhwelo. The Indian armed force didn't permit the friends and family to perform last customs and ceremonies for the dead. All the dead bodies were dumped inside the town chief’s house and were torched to remains alongside different houses and granaries.The ladies and kids who had fled to the wilderness to dodge the repulsiveness and torment of Indian Army returned the following morning to locate the entire town torched to cinders. Lt. Thitu who barely got away from the execution was found by his better half Mrs. Rhiitariih with three cuts on the neck, stomach cut and digestive organs tossed out. He cited â€Å"Love, tell my cherished kids the penance I have borne for them and I am holding back to bite the dust in your lap with some water† and in the wake of drinking, he inhaled his last. Another casualty Lt. Zasituo, voyaging P astor, was additionally discovered practically dead with different wounds on his chest and neck.Not long after, he passed on. At that point the frightened ladies and youngsters without any methods essentially secured the dead bodies with mud and left for the wildernesses dreading the Indian armed force may turn up whenever. For a considerable length of time together, the survivors meandered in the profound wilderness without appropriate food and haven. The wild berries and products of the wilderness were their lone food and methods for endurance. The main solace and consolation they could provide for one another was the information on brilliant penances made by their men people. The wild creatures and feathered creatures of the wilderness were their solitary allies, other than themselves.In extraordinary states of hardships and trouble, a lot progressively valuable lives were lost. The Naga Army at that point acted the hero. They were given food, haven and assurance. Indeed, even to day, the bad dreams and disaster of the occurrence despite everything stay new in the psyche of the survivors. In this long scattering and mass migration, the survivors entered Burma and remained with the Naga Army in their camp at Sathi where Gavin Young of London Observer met them in the later piece of 1961. In his book â€Å"Indo-Naga War†, page 29-30, he composed that when he met the survivors, there were just a terrible thirty people.They spent their lives in the wild for two and half years. In 1963, town re-foundation occurred however typical life couldn't be reestablished for a long time. The individuals of Pochury have since the time watched sixth September as â€Å"BLACK DAY† in memory of each one of the individuals who had endured and set out their lives for the more prominent brilliance and opportunity of the Nagas. It is a day of misery and a day to affirmation of the penances made by the saints. Khuosatho Nyusou, President, P1ochury Hoho Kohima (PHK). (Pol iteness: Pochury Students Union. )|

Reflection and Practice Gibb’s Model Samples †MyAssignmenthelp.com

Question: Examine about the Reflection and Practice Gibbs Model Guides. Answer: Presentation: Gibbs model aides reflection when dealing with a circumstance. This model is useful in separating the circumstance into occasions and individual reactions. This reflection cycle is the connection between gaining from an earlier time and future arranging. Gibbs intelligent cycle prompts medical attendants to think in an orderly manner about each period of the training. Experience is significant for learning, anyway attendants should think continually and ponder it, for development practically speaking. It is additionally helpful in understanding specific frequency. Consequently, it is useful to comprehend the occurrence in better manner and there are odds of enlargement in the quality next time. Situation: This paper is about Mr. X (65 yrs.). He was admitted to the ward two days before because of indications of dementia and his unpredictable and unsuitable conduct towards his relatives and companions. I had been throughout the previous 5 years as a private medical attendant in the ward for mental ailments. I generally used to show up busy working by 7 a.m. I used to help more seasoned patients in the ward in their every day exercises of the morning meeting. These incorporate utilization of toilets and washroom. At the beginning of today likewise, I was occupied with these standard assignments. While playing out these exercises, I experienced Mr. X. Preceding conversing with him; I had gone through around 30 minutes in the ward. During this time, he was exceptionally peaceful and lying on his bed. I asked him, regardless of whether he required help for the utilization of the can and restroom. Rather than reacting to me, he began mishandling me and my partners. Out of nowhere, he got up from his bed and began talking boisterously and he was moving his both the hands quick. He was attempting to hurt me genuinely, anyway I got away securely. From the earlier night he had been enquiring about his better half. Notwithstanding, his significant other kicked the bucket three years back because of a coronary episode. He was over and again saying that staff individuals were not permitting him to meet his significant other. In this negative circumstance likewise I kept my nerves cool and with the help of ranking staff part, I persuaded him. I hold his submit my hand and mentioned him to quiet down. I mentioned senior individuals from my staff to converse with him and shellfish down him. I did this by listening cautiously to him. I didn't irate with him. I let him know, I needed to support him. I requested that he be progressively agreeable. He didn't need me to remind his better half, so I concurred. Reflection: Feelings At first, I had complex sentiments during the occurrence. I was reevaluating about my language, regardless of whether I had utilized wrong words. I felt humiliated because of that rate and I was vexed. I felt that, I had upset him. I was thinking, he was showing disturbed conduct because of me. Because of this episode, different patients in the ward additionally become upset. Different patients may be believing that I didn't give legitimate treatment to Mr. X. These patients may be thinking, I utilized egotistical words to Mr. X, henceforth he got disturbed and carried on as he did. My emotions became troubling in light of the fact that, I made him review his dead spouse. I was scared at the hour of occurrence; anyway I resisted the urge to panic around then and dealt with the circumstance in an exceptionally proficient way. I didn't leave the patient in his disturbed conditions (Roberts and Dyer, 2004). My emotions were acceptable and I was fulfilled on the grounds that I picked up trust in taking care of such a circumstance viably. My emotions were totally unique in relation to the sentiments toward the beginning of episode. A definitive was certain. Mr. X consented to utilize the can and restroom. Fruition of these morning meeting exercises of Mr. X was significant on the grounds that it can assist with making him stable and to improve his solace level (Baillie, 2005). Assessment: The beneficial thing about this experience was, I followednursing norms and qualities in an exceptionally proficient and moral manner. The terrible thing about the occurrence was, I got abuse from the patient. Patients acting mischievously withnursing and medicinal services staff, who were taking such a large number of endeavors for physical and mental prosperity of the patients. Another terrible thing about the occurrence was, different patients in the ward likewise got upset because of this rate. Examination: I could detect that, others may have dealt with this occurrence with various methodology. My methodology of taking care of this occurrence was law based way. I listened his emotions cautiously and took him in certainty. Others may have taken care of this occurrence in increasingly absolutist manner. They may have taught him to play out his standard exercises like latrine and washroom use. Such sort of demeanor may have misrepresented the circumstance. This may have lead to unsettling influence to different individuals in the ward. Different individuals from the staff were responded emphatically to this circumstance. My seniornursing staff and ward kid had helped me to quiet down Mr. X and to take him to can and washroom use. (Corrigan et al., 2005). Other staff individuals had attempted to energize him and furthermore had attempted to improve his conviction innursing and medicinal services staff. They had revealed to him that nursing staff implied for prosperity of the patients and t hey would do nothing destructive to him. Subsequently, he ought to have total trust on nursing staff for his quick recuperation (Anthony, 2000x; May, 2000). For Mr. X, chances of interior recuperation process were unimportant in light of the fact that because of his age he would not have the option to comprehend results of his conduct. Redundant relationship between patients experience and social conditions, including endeavors and uplifting demeanor of me and other staff individuals from the ward, had helped in improving state of Mr. X. Wellbeing and nursing staff were certain about the improvement of the patients with mental ailment like Mr. X. This idealistic methodology of the staff individuals had helped in expanding certainty level and trust in recuperation of Mr. X (Roberts Wolfson, 2004; Repper and Perkins, 2003). It would be useful for me handle such conditions in increasingly compelling and productive manner. I would develop myself in giving intercession to such intellectually sick patients by conversation with senior individuals from the medical clinic, concentrating more examinations and including myself in the quantity of studies. This frequency had helped me to build my certainty level in taking care of more instances of mental and mental patients. I would accomplish more research on comparative kinds of cases and study it cautiously. I would talk about these cases with my seniors to get their view on these cases. I would offer types of assistance to such patients by incorporating information, aptitudes, research, procedures and experience. Finish of reflection: From this occasion it was clear that there ought to be arrangement of isolated ward for the intellectually sick patients. It would be useful in a tough situation to different patients in the ward. Morevoer, there ought to be enlistment of particular staff individuals in the administration of intellectually sick patients. It was acceptable that, me and other staff individuals had overseen instance of Mr. X in legitimate manner to quiet him. Else, it would have been troublesome situation to control in the ward, in the event that he would have not been constrained by the current staff individuals. I, alongside other staff individuals had dealt with this instance of Mr. X on the grounds that as indicated by our emergency clinic wellbeing approach we ought not leave quiet on his/her condition. It is duty of each staff part to comply with the standards and guidelines of the strategy of the medical clinic. The board ought to improve this strategy by revising the emergency clinic approach. I n the medical clinic approach, there ought to be arrangement for welcoming master from different emergency clinics if there should be an occurrence of crisis (Goel, 2010; Peters, 2016). Activity plan: I ought to likewise take a shot at improving my appearance capacities. I should rehearse appearance in number of cases. Reflection and evaluation of the nursing practice would be useful to manage the unfavorable conditions. It would be useful for me to confront any case decisively. Next time, I would ask individual patients in the ward to direction such unsuitable conduct of the patients. Scarcely any patients may be agreeable to impart their sentiments to different patients when contrasted with the attendant and human services staff. It would be useful for the attendant like me to quiet the patient and diminish trouble on me. Understanding: I dealt with the circumstance successfully and took care of the circumstance as indicated by rules and guidelines (Grace, 2013). I dealt with this circumstance, without a specialist in the field on the grounds that in our emergency clinic there is no master for mental and mental wellbeing. This episode showed me a great deal and it gave me opportunity to evaluate myself in taking care of unfriendly conditions. I comprehended that these experience can not be picked up in homeroom instructing. Or maybe such continuous encounters can instruct. I discovered that my productive correspondence, uplifting disposition and all encompassing methodology helped me to deal with this condition (Thresyamma, 2005). However, I went over such rates just because, I gained aptitudes and capacity to deal with such unfavorable conditions structure my guide. I used to watch him distinctly while dealing with such conditions. Because of medical clinic approach, there was no different course of action accessible for the administration of intellectually sick patients. Consequently, these patients ought to be conceded in a similar ward alongside different patients. Most significant taking in for me from this occurrence was that, correspondence is most significant factor for the administration of the patients with dementia. Making acknowledgment is significant for such patients on the grounds that these patients used to overlook the greater part of the things. Mindfulness had helped me to manufacture correspondence with the patients. I would rehearse mindfulness more since it is clear that mindfulness is valuable instrument for overseeing intellectually sick patients. For the administration of intellectually sick patients, it is a lot of required to comprehend their emotions and conduct. Mindfulness would supportive to get knowledge of the intellectually sick patients and reflect emphatically as indicated by their b

13 rules you did not learn in school

13 rules you did not learn in school Here are some basic rules that children should be learning in  school, but unfortunately dont. Not all of these have to do with academics.Rule #1: Life is not fair. Get used to it. The average teen-ager uses the  phrase, Its not fair 8.6 times a day.Rule #2: The real world wont care as much about your self-esteem as much  as your school does. Itll expect you to accomplish something before you  feel good about yourself. This may come as a shock. Usually, when inflated  self-esteem meets reality, kids complain its not fair. Rule #3: Sorry, you wont make $40,000 a year right out of high school.  And you wont be a vice president or have a car phone either. You may even  have to wear a uniform that doesnt have a label.Rule #4: If you think your teacher is tough, wait until you get a boss. He  doesnt have tenure, so he tends to be a bit edgier. When you screw up, hes  not going to ask you how you feel about it.Rule #5: Flipping burgers is not beneath your dignity. Your grand-parents  had a different word of burger flipping. They called it opportunity. They  werent embarrassed making minimum wage either. They would have been  embarrassed to sit around talking about Kurt Cobain all weekend.Rule #6: Its not your parents fault. If you screw up, you are  responsible. This is the flip side of Its my life, and Youre not the  boss of me, and other eloquent proclamations of your generation. When you  turn 18, its on your dime. Dont whine about it, or youll sound like a  baby boomer.Rule #7: Your school may have done away with winners and losers. Life  hasnt. In some schools, theyll give you as many times as you want to get  the right answer. Failing grades have been abolished and class  valedictorians scrapped, lest anyones feelings be hurt. Effort is as  important as results. This, of course, bears not the slightest resemblance  to anything in real life.Rule #8: Life is not divided into semesters, and you dont get summers  off. Not even Easter break. They expect you to show up every day. For eight  hours. And you dont get a new life every 10 weeks. It just goes on and on.  While were at it, very few jobs are interesting in fostering your  self-expression or helping you find yourself. Fewer still lead to  self-realization.Rule #9: Television is not real life. Your life is not a sitcom. Your  problems will not all be solved in 30 minutes, minus time for commercials.  In real life, people actually have to leave the coffee shop to go to jobs.  Your friends will not be as perky or pliable as Jennifer Aniston.Rule #10: Be nice to nerds. You may end up working for them. We all  could.Rule #11: Smoking does not make you look cool. It makes you look moronic.  Next time youre out cruising, watch an 11-year-old with a butt in his  mouth. Thats what you look like to anyone over 20. Ditto for expressing  yourself with purple hair and/or pierced body parts.Rule #12: You are not immortal. If you are under the  impression that living fast, dying young and leaving a b eautiful corpse is  romantic, you obviously havent seen one of your peers at room temperature  lately.Rule #13: Enjoy your youth time while you can. Sure parents are a pain, school is a  bother and life is depressing but someday youll realize how wonderful it  was to be a kid. Maybe you should start now.By Charles J. Sykes Printed in San Diego Union Tribune September 19, 1996

Social and Ethical Technology Concerns-Biotechnology

Social and Ethical Technology Concerns-Biotechnology Biotechnology is technology based on living organisms. It harnesses cellular and biomolecular processes to develop technologies or products which are immensely useful to mankind. Biotechnology modifies living organisms according to our needs. Humans have employed this tool in the fields of agriculture, food industry and medicine for more than 6000 years. We see numerous biotechnological applications in our every-day life. Preparation of food products like bread, cheese and preservatives in dairy products are all outcomes of this great scientific tool (Biotechnology Industry Organization). Modern biotechnological research has created wonders for mankind. It has provided products to combat rare diseases, vaccines for untreatable diseases, food for the hungry and a safer, cleaner environment to our planet. Biotechnology has played a vital role in reducing the green house effect and in producing a cleaner planet. Because of the worldwide economic and environmental concerns regarding the use of petro-chemicals, extensive research has been conducted on residual biomass. Significant progress has been made in the field of lignocellulose biotechnology. Lignocellulosic waste materials have been used in the production of bio-fuels, enzymes, chemicals, the pulp and paper, animal feed and composites (Iqbal, 2013). Moreover, algal biomass has received huge attention in producing biofuels due to their relatively high growth rate, greatShow MoreRelatedEssay about Sci 115 Week 8 Assignment 2 Gene Technology1132 Words   |  5 PagesSCI 115 WEEK 8 ASSIGNMENT 2 GENE TECHNOLOGY To purchase this visit here: http://www.activitymode.com/product/sci-115-week-8-assignment-2-gene-technology/ Contact us at: SUPPORT@ACTIVITYMODE.COM SCI 115 WEEK 8 ASSIGNMENT 2 GENE TECHNOLOGY SCI 115 Week 8 Assignment 2 - Gene Technology Gene technology carries with it social and ethical implications—many of which engender personal views and discussion. Select one (1) of the following biotechnology topics to write about: †¢ GeneticallyRead MoreModern Food Biotechnology, Human Health And Development : An Evidence Based Stud1561 Words   |  7 PagesModern Food Biotechnology, Human Health and Development: An Evidence-Based Stud Food security is a recurrent issue in modern society due to the continual increase in human population. 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Nature vs. Society Wordsworths Romantic Poetry - 1646 Words

Nature Vs. Society: Wordsworth’s Romantic Poetry Over time, poetry has changed and evolved in its sense of the word nature. In its beginnings the idea of nature or natural was seen as negative and evil. However, in more recent times due to the era of Romanticism, nature in poetry is viewed in a positive and even beautiful light. William Wordsworth was a poet who wrote his poetry with a romantic attitude. Furthermore Wordsworth wrote specifically the poems â€Å"We Are Seven† (WAS) and â€Å"Three Years She Grew† (TYSG) in a style that showcased the superiority of nature over society. â€Å"We Are Seven† and â€Å"Three Years She Grew† portray a romantic attitude in their works, additionally the values placed on the natural world over the societal world†¦show more content†¦The second poem â€Å"Three Years She Grew† has the same theme of death however, is from a different perspective. Generally, the poem is about a man who share s a love for a woman although her life is short lived. The poem is full of compliments towards nature, and to be associated is seen to be of the upmost regards. In the first stanza, the audience is hearing about how nature feels about this now young three-year-old girl also known as Lucy. Nature is ultimately personified almost as a mother figure or further Mother Nature in which â€Å"adopts† the young girl. Then Nature said, â€Å"A lovelier flower On earth was never sown; This Child I to myself will take; She shall be mine, and I will make A Lady of my own. (Wordsworth, TYSG, 2-6) In this verse, nature is choosing the young girl for it’s own, which in the natural world can be viewed as one of the best things that can happen to an individual. Also, nature marks the girl as â€Å"Lady† which invokes a class status, that of upper class. Lucy will be a natural lady, and thus does not need society’s confirmation of this, furthermore contrasting nature to society in social recognition. The second stanza also compares nature to society in the use of social laws. â€Å"Both law and impulse: and with me†¦/ †¦[s]hall feel an overseeing power/ [t]o kindle or restrain†. (Wordsworth, TYSG, 8-12) InShow MoreRelatedThe Romantic Movement Of William Wordsworth And Samuel Taylor Coleridge Essay1427 Words   |  6 Pages INTRODUCTION The Romantic Movement was introduced around1820 to America; this movement overlapped with the period of national development. Romantic philosophies looked at art as spiritual elements of nature and used metaphors as an inspiration. Nature was a significant theme for the duration of this period since supernatural things began in our imagination. Many sonnets were created using freedom of thought however these writings soon shifted to an imagination method (natural surroundings) fromRead MoreAnalysis Of Daffodils By William Wordsworth2381 Words   |  10 Pagesdedicated to a return to nature within literature and art. It was a confident reaction to the industrial revolution, in which man-made objects became the life of soul of the everyman’s everyday life. The great artists and poets and authors of the time began to create works filled with passion and emotion and all interpreted from the themes within nature. During this period, authors and artists alike found inspiration in things such as flowers, for example William Wordsworth’s classic entitled ‘daffodils’Read MoreThe Romantic Period Stressed Instincts, Affection, and Love1422 Words   |  6 PagesThe Romantic period or Romanticism was a literary movement that had sprung in England in late 18th century. The rise of the movement was precipitated by the issuing of Lyrical Ballads by William Wordsworth and Samuel Taylor Coleridge .The movement came to cope up with the general tenors of the Industrial and Enlightenment age .It stressed on things like instincts, affection , love,the heart over the head .It came also to celebrate such things as mysticism and the natural world.Romanticism alsoRead MoreRomanticism versus Neoclassicism2775 Words   |  12 PagesNeoclassical and Romantic movements cover the period of 1750 to 1850. Neoclassicism showed life to be more rational than it really was. The Romantics favoured an interest in nature, picturesque, violent, sublime. Unlike Neo_cla ssicism, which stood for the order, reason, tradition, society, intellect and formal diction, Romanticism allowed people to get away from the constrained rational views of life and concentrate on an emotional and sentimental side of humanity. In this movement the emphasis was

Association of Human papillomavirus and Breast Tumours Free Essays

string(215) " apoptosis of abnormal cells, whereas E7 inactivates Rb \(retinoblastoma\) function, which results in abnormal cell proliferation and disturbs the normal cell cycle regulation \(Wang, 2007; WHO, 2006; Mera, 1997\)\." Abstract: Cancer arises due to abnormal changes or mutations, in the genes responsible for regulating the growth of cells. Human papillomavirus (HPV) is considered the aetiological agent for many cancers including cervical cancer. HPV causes disruption and loss of some of the viral genes such as L1 and L2 genes and also increases the expression of the early genes. We will write a custom essay sample on Association of Human papillomavirus and Breast Tumours or any similar topic only for you Order Now Several studies have addressed a relationship with HPV and breast cancer, as different HPVs have been identified. Most of the studies were successful in finding evidence in correlation of HPV 6, 11, 16 and 18 in invasive ductal breast carcinoma by using different techniques including DNA extraction and PCR, however other studies achieved low positivity or negative result. The aim of this study was to find out the association of HPV and Breast cancer. DNA was successfully extracted from archived breast tissue samples using DNA extraction method. This DNA sample could be amplified by using PCR to find HPV genome specifically targeting E1 gene. This is an ongoing work by the supervisors of the project to try and detect HPV genome in breast cancer, if successful a vaccine could be developed against various strains of HPVs worldwide and it could save many lives. Keywords:Human papillomavirus, DNA Extraction, Breast cancer, Vaccine, PCR, 1. Introduction: 1.1. Breast cancer: Breast cancer is a malignant tumour that originates in the breast tissue, mainly from the inner lining of milk ducts or the lobules that supply milk to the ducts, cancers that initiates from ducts are called ductal carcinomas and those originating from lobules are called as lobular carcinomas. Cancer occurs due to abnormal changes or mutations, in the genes responsible for regulating the growth of cells (Sariego, 2010). The change in the genetic information causes a cell to no longer carry out its function properly (Almeida Barry, 2010). The following figures show the two types of cancers Benign and Malignant. (Almeida; Shela, 2010) Figure: 2 malignant tumours Figure 1 and 2: above shows benign vs. malignant cancers. (a) A benign tumour is a mass of cells that remains within the tissue in which it originally developed. (b) The invasion of cancer cells into surrounding tissues is the hallmark of a malignant tumour. Malignant cells may break free from the tumour and travel to other locations in the body through the process of metastasis (Almeida Barry, 2010). 1.2. Epidemiology: Breast cancer is one of the main health problems worldwide (Bao, 2011) and which resulted http://en.wikipedia.org/wiki/Breast_cancer – cite_note-WHO_WCR-2 458,503 deaths in 2008 worldwide out of which 13.7% are of cancer deaths in women and it is about 100 times more common in women than in men (Veto, et al., 2009). The table below shows how females are susceptible to breast cancer at different ages for example there is 1 in 8 risk of developing breast cancer in females in the U.K in lifetime. Table 1: Shows estimated risk of developing breast cancer by age, females, UK, 2008 UK, 2008 Adopted from: www.cancerresearch.uk http://info.cancerresearchuk.org/cancerstats/types/breast/riskfactors/ Date accessed: 20/01/11 The table 2 below shows that more deaths happens in females due to breast cancer than males as it can be seen from the table only 69 males died in 2008 in compare to 12,047 females. Table 2: Shows the number of deaths and mortality rates in the UK in 2008. Adopted from: www.cancerresearch.uk http://info.cancerresearchuk.org/cancerstats/types/breast/mortality/#age Date accessed: 20/01/11 The figure below shows the incidence and mortality rates from female breast cancer in EU countries. As it can be seen from the table Belgium has the highest rates of incidence in female breast cancer. Figure 1 above is a graph of incidence and mortality rates in EU. Adopted from: www.cancerresearch.uk http://info.cancerresearchuk.org/cancerstats/types/breast/incidence/ Date accessed: 20/01/11 1.3 . Breast cancer Pathophysiology: 1.3.1 Aetiology: Some of the suspected aetiological factors which influence the cases of breast cancer are family history, obesity, age, oral contraceptives and alcohol. Family history: A woman who has a family member with breast cancer increases double the risk of getting breast cancer in compare to a woman with no family history (Lancet, 2001). Obesity: obesity increases the risk of postmenopausal breast cancer by up to 30%, since levels of hormones rises with excess body fat such as oestrogen and insulin these are the common features of cancers. Age: older women are at higher risk. Particularly women aged 50-69 are most at risk, predominantly those with a late menopause. Oral contraceptives: increases the risk by approximately a quarter but since people who uses are commonly younger women, therefore the risk is fairly low. Alcohol: drinking alcohol as less as one alcoholic drink each day increases the risk of breast cancer by around 12%. (Cancer Research U.K, 2008) Some other factors include: Lesions to DNA such as genetic mutations. There is link between mutations that can lead to breast cancer and oestrogen exposure, found out by carrying out experiments. Another factor is when a body fails to carry out immune surveillance; it is a theory in which the immune system gets rid of malignant cells throughout one’s life. Other factor is inherited defects in DNA repair genes, such as †BRCA1†, †BRCA2† and †TP53† (Adams, et al., 2011). Figure 2 above shows the percentage of different genes with associated risk. Figure adopted from: Wooster and Webber, (2003) Date accessed: 12/04/11 Moreover according to many authors there is a potential link between the HPV and breast cancer. 1.4. Human papillomavirus and Cancer: HPV genome is normally found in the cytoplasm of infected tissues however, the DNA of HPV types that cause cancer are integrated into the host genome. HPV causes disruption and loss of some of the viral genes for example (L1 and L2 genes) and also increases the expression of the early genes (Wang, 2007; Mera, 1997). Oncoproteins E5 interacts with MHC I and prevents its transport to the cell surface therefore infected cells escapes the immune system consequently allowing the virus to establish persistent infections and thus progressing to cancer. E6 targets p53 for degradation and therefore prevents apoptosis of abnormal cells, whereas E7 inactivates Rb (retinoblastoma) function, which results in abnormal cell proliferation and disturbs the normal cell cycle regulation (Wang, 2007; WHO, 2006; Mera, 1997). You read "Association of Human papillomavirus and Breast Tumours" in category "Essay examples" P53 and Rb are tumour suppressor genes which stop tumours from developing (Mera, 1997). Incorporation of virus into host cell increases and sustains the growth of both virus and the host cell, thus resulting in the alteration of infected host cells into malignant cells (Mera, 1997; Wang, 2007) and ultimately invasive cancer. Figure 3 above shows different genes in HPV. Adopted from: Symptoms of HPV 2010 symptomsofhpv.net/113/hpv-16/ Date accessed: 07/04/11 Table 9 below shows the function of different genes within the HPV virus: Gene/RegionFunction E1/E2Code for proteins which control the function of E6 and E7 genes. E4Function largely unknown but may control virus release from cell. E5Codes for a hydrophobic protein which enhances immortalisation of the cell. E6Codes for proteins which inhibit negative regulators of the cell cycle .E6 products inhibit p53 which is a transcription factor for apoptosis (programmed cell death). E7Codes for products whichbind to the retinoblastoma tumour suppressorproteins thereby permitting the cell to progress through the cell cycle in the absence of normal mitogenic signals. L1/L2Code for structural proteins and formation of complete virus particles. LCRNecessary for normal virus replication and control of gene expression. Adopted from: Eurocytology http://www.eurocytology.eu/static/eurocytology/eng/cervical/LP1ContentMcontA1.html Date accessed: 19/03/11 The HPV (human papillomavirus) is a member of the papillomaviridae family and has a double stranded circular DNA genome (Wang, 2007). These viruses are small in size with 8kbp-long DNA genome and have no envelope (WHO, 2006). HPV genome contains early (E) and the late genes (L) which codes for early proteins (E1-E7), late proteins (L1 and L2) and a non coding long control region (LCR) (WHO, 2006; Mera, 1997; Govan, 2008). 1.4.1. High risk and low risk HPV types: There are more than one hundred different HPV types that have been discovered (WHO, 2006) and these are divided into high risk and low risk types. HPV 16, 18, 31 and 45 are some high risk HPV types associated with most of the cancer, while HPV 6 and 11 are low risk non-oncogenic HPV types (Brown, et al., 2005; Govan, 2008). Table3: the following table shows some high risk, low risk and potentially risk HPVs. ClassificationHPV types High-risk16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 Low-risk82, 6, 11, 40, 42, 43, 44, 54, 61, 70 Potentially high –risk26, 53 Source: Govan (2008) HPV 6 and 11 are linked with up to 90% genital warts (Von Krogh, 2011), nevertheless after the examination of 55 genital wart samples from Slovenia, using polymerase chain reaction (PCR), the authors concluded that HPV 6 and 11 genotypes were detected in 96.4% of genital warts patients (Potocnik, et al., 2007). 1.5. Signs and Symptoms: Changes that could arise due to a breast cancer are: A change in the size or shape of a breast A lump or thickening in an area of the breast Dimpling of the skin A change in the shape of the nipple, particularly if it turns in, sinks into the breast or becomes irregular in shape A blood stained discharge from the nipple (Dixon, 2005; Breast cancer, 2008). Figure 3a: above shows the symptoms of breast cancer Source: Healthbase (2008) http://blog.healthbase.com/2008_09_01_archive.html Accessed date: 11/04/2011 Normal anatomy of the breast: Female breast anatomy The structure of female breast is mainly made up of fat and connective tissue, but also contains milk ducts, lymph nodes, blood vessels and structures known as lobes and lobules (Rosen, 2009). Figure 4 above shows normal anatomy of breast tissue. The above figure adopted from: Mayoclinic (2009) http://www.mayoclinic.com/health/breast-cancer-early-stage/BC00001 Date accessed: 8 April 2011. Lobules and ducts Every breast has 12 to 20 lobules that protrude from the nipple and holds small alveoli; the lobules are connected together by a network of thin ducts (Rosen, 2009). Figure 5 above shows different parts in the female breast The above figure adopted from: Mayoclinic (2009) http://www.mayoclinic.com/health/breast-cancer-early-stage/BC00001 Date accessed: 8 April 2011. Stromata Spaces around the lobules and ducts are filled with fatty tissue, ligaments and connective tissue (stromata). The size of the breast is determined by the amount of fat it contains, the breast tissue is also sensitive to cyclic changes in hormone levels (Rosen, 2009). Figure 6 above shows the position of stromata in female breast. The figure adopted from: Mayoclinic (2009). http://www.mayoclinic.com/health/breast-cancer-early-stage/BC00001 Date accessed: 8 April 2011. Muscles Breasts are muscle free tissues, muscles lie beneath the breasts separating them from the ribs (Rosen, 2009). The above figure adopted from: Mayoclinic (2009) http://www.mayoclinic.com/health/breast-cancer-early-stage/BC00001 Date accessed: 8 April 2011. Arteries and capillaries Blood supply all the essential nutrients and oxygen to the breast tissue through arteries, capillaries and small blood vessels (Rosen, 2009). Figure 8 above shows the position of capillaries and arteries in and around the breast. Figure adopted from: Mayoclinic, (2009) http://www.mayoclinic.com/health/breast-cancer-early-stage/BC00001 Date accessed: 8 April 2011. Lymph nodes and lymph ducts The lymphatic system contains blood vessels, lymph ducts and lymph nodes that helps fight infection, lymph nodes are present behind the breastbone, under the armpit and in other parts of the body engulfs harmful substances that are in the lymphatic system and safely get rid of them (Rosen, 2009; Mayoclinic, 2009). Figure 9 above shows the position of the lymph nodes and lymph ducts. The above figure adopted from: Mayoclinic, 2009 http://www.mayoclinic.com/health/breast-cancer-early-stage/BC00001 Date accessed: 8 April 2011. 1.4. Different types of Breast cancers: There are different types of breast cancer for example ductal and lobular and it depends on the type of tissue that it is derived from. Table 3.1 below shows the list of different types of breast cancer: DCIS – ductal carcinoma in situ LCIS – lobular carcinoma in situ Invasive ductal breast cancer Invasive lobular breast cancer Inflammatory breast cancer Paget’s disease Breast cancer in men The following figures show some of the main types of the cancer that begins in different areas of the breast for example the ducts, the lobules, or in some cases, the tissue in between. These figures also show the different types of breast cancer, including non-invasive, invasive, metastatic and recurrent breast cancers. a. Ductal Carcinoma in situ (DCIS) Range of Ductal Carcinoma in situ (DCIS) Figures: 10 and 11 above show normal breast with non–invasive ductal carcinoma in situ (DCIS) in an enlarged cross–section of the duct. Adopted from: Breast cancer (2008) http://www.breastcancer.org/symptoms/types/dcis/diagnosis.jsp Accessed date: 20/01/2011 Breast profileABCDEFG DuctsLobulesDilated section of duct to hold milkNippleFatPectralis major muscleChest wall/ rib cage EnlargementDuctal cancer cellsNormal lobular cellsBasement membraneLumen ( centre of duct) Table 4: shows the annotation of the above figures (Trentham-Dietz, et al., 2011) b. Lobular Carcinoma in situ (LCIS Figure: 12 above shows normal breast with lobular carcinoma in situ (LCIS) in an enlarged cross–section of the lobule. Adopted from: Breast cancer (2008) http://www.breastcancer.org/symptoms/types/ilc/tests/diagnosing.jsp Accessed date: 20/01/2011 Table 5 shows the annotation of the figure 12. Breast profileABCDEFG DuctsLobulesDilated section of duct to hold milkNippleFatPectralis major muscleChest wall/ rib cage EnlargementNormal Ductal cellsLobular cancer cellsBasement membrane (Trentham-Dietz et al., 2011) a.Invasive Ductal Carcinoma (IDC) Figure 13 above shows normal breast with invasive ductal carcinoma (IDC) in an enlarged cross–section of the duct. Adopted from: Breast cancer (2008) http://www.breastcancer.org/symptoms/types/ilc/tests/diagnosing.jsp Accessed date: 20/01/2011 Breast profileABCDEFG DuctsLobulesDilated section of duct to hold milkNippleFatPectralis major muscleChest wall/ rib cage EnlargementNormal duct cellsductal cancer cells breaking through the basement membrane Basement membrane Table 6 shows the annotation of the figure 13. (Trentham-Dietz, et al., 2011) c. Invasive Lobular Carcinoma (ILC) The above figure 14 shows normal breast with invasive lobular carcinoma (ILC) in an enlarged cross–section of the lobule. Adopted from: Breast cancer (2008) http://www.breastcancer.org/symptoms/types/ilc/tests/diagnosing.jsp Accessed date: 20/01/2011 Table 7 shows the annotation of the figure 14. Breast profileABCDEFG DuctsLobulesDilated section of duct to hold milkNippleFatPectralis major muscleChest wall/ rib cage EnlargementNormal cellsLobular cancer cells breaking through the basement membraneBasement membrane (Trentham-Dietz, et al., 2011) Following are some examples of non-invasive cell growths: d. Non-Invasive Cell Growth Subtypes – Solid Figure: 15 shows A cancer cells B basement membrane Adopted from: Breast cancer (2008) Accessed date: 20/01/2011 e.Non-Invasive Cell Growth Subtypes – Cribriform Figure: 16 above shows (A) cancer cells (B) basement membrane (C) lumen (centre of duct) Adopted from: Breast cancer (2008) Accessed date: 20/01/2011 f. Non-Invasive Cell Growth Subtypes – Papillary Figure: 17 above shows (A) cancer cells (B) basement membrane (C) lumen (centre of duct) Adopted from: Breast cancer (2008) Accessed date: 20/01/2011 g. Non-Invasive Cell Growth Subtypes – Comedo Figure: 18 above shows (A) living cancer cells (B) dying cancer cells (C) cell debris (necrosis) Adopted from: Breast cancer (2008) Accessed date: 20/01/2011 h. Vascular and Lymphatic Invasion Figure: 20 above shows normal breast with cancer cells invading the lymph channels and blood vessels in the breast tissue Adopted from: Breast cancer (2008) Accessed date: 20/01/2011 Table 8 shows the annotation of the above figure Breast profileABCDEF Blood vesselsLymphatic channels EnlargementNormal duct cellscancer cellsBasement membraneLymphatic channelBlood vesselBreast tissue 1.4. Diagnostic tests: Diagnosis of the breast cancer incorporates x-rays and screening tests and following are some of the important diagnostic tests that can be carried out before and after symptoms of breast cancer. Tests: Mammogram: A mammogram is the main screening test for asymptomatic patients who are over the age of 40 as well as for symptomatic adult patients (Bao, 2011).This test has a high sensitivity and specificity (Banks, 2004). If a mammogram does not find out an abnormality in patients with a clinically detected breast mass, additional imaging ultrasound and/or MRI should be carried out for further evaluation (Bao, 2011). Outcome: The results are indicative of malignancy include: an irregular speculated mass, clustered micro-calcifications, and linear branching calcifications (Banks, 2004; breast cancer, 2010). The above figure 21 shows how mammography is carried out. Figure 21 adopted from: Breast cancer (2010) http://www.breastcancer.org/symptoms/testing/types/mammograms/ Accessed date: 02/04/2011 Breast Ultrasound: Ultrasound sends high-frequency sound waves through the breast and changes them into images on a screen. The ultrasound technician places a sound-emitting probe on the breast to carry out the test and there is no radiation involved (Matsuzaki, et al., 2010). Outcome: The results are indicative of malignancy include: a hypo echoic mass, an irregular mass with internal calcifications, and enlarged auxiliary lymph nodes (breast cancer 2010; Moss, 1999). The above figure 22 shows how ultrasound is carried out. adopted from: Breast cancer (2010) http://www.breastcancer.org/symptoms/testing/types/ultrasound.jsp Accessed date: 02/04/2011 Breast MRI: MRI uses magnets and radio to produce detailed cross sectional images of the inside of the body. MRI screens high-risk women (breast cancer, 2010). The Sensitivity is 88% to 91%. Specificity is about 67% (Bluemke, 2004). Outcome: The results are indicative of malignancy include: a heterogeneously enhancing area and significant architectural distortion (Bluemke, 2004). The above figure 23 shows how MRI is carried out. adopted from: Breast cancer (2010) http://www.breastcancer.org/symptoms/testing/types/mri/ Accessed date: 02/04/2011 Biopsy: There are different types of biopsy techniques and among these techniques Fine needle aspiration (FNA) is the least invasive procedure and has high sensitivity and specificity (Dayal, et al., 2011). FNA is good for quick diagnosis of malignancy. Nonetheless, core biopsy is generally favoured, as it effectively differentiates between pre-invasive and invasive disease and is less chance getting inadequate sampling (Dayal, et al., 2011). Outcome: Invasive ductal carcinoma is responsible for almost 80% of all breast cancers, cords of tumour cells among associated glandular formation, which make varying degrees of fibrotic response. Whereas invasive lobular carcinoma, small tumour cells that invade past the basement membrane of the lobules and form an â€Å"Indian file† between collagen bundles, usually appears as well-differentiated tumour cells that display tubule formation (Dayal, et al., 2011). 1.5. Aims and Objectives: The aim of this project was to evaluate the association of Human papillomavirus (HPV) and breast cancer, additionally to collect the studies that support the presence of HPV DNA in patients with breast lesions worldwide. The archived samples diagnosed with breast carcinoma, will be used to extract the DNA by DNA Extraction method which could be further used for amplifying this DNA using PCR to detect HPV genome. This will ascertain the role of this virus in the pathogenesis of breast cancer and will also help the scientist for further investigation of this virus on biology of cancer. The following is the methodology of my project as how I carried out the experiment and extracted the DNA. 2. Methodology: The methodology incorporates materials and method, health and safety, ethical issues and statistical analysis. 2.1. Method and Materials: The following table 9 shows the materials that have been used to extract the DNA. Materials Measurements Universal tubes 20 ml, 5ml Epindorf tube 1.5 ml, 500ul Gilson pipetts 2x 5- 50ul, 2x 0.1 – 2.5 ul,2x 100 – 1000ul, 1x 20 – 200ul Dry heat block ( incubator) Vortex Waterbath ( 37c) Centrifuge and microfuge 70% of alcohol to avoid contamination and spray bottle Ice box 10. Thermometer( to measure the temperature) 11. Spectrophotometer( OD reader) 12. Pipett tips 13. Tissue and Cell LysisSolution 600ul (60ml) 14. Proteinase K4ul ( 200ul) 15. RNase A2ul (400ul) 16. Protein Precipitation Reagent300ul (60ml) 17. Isopropanol1ml (2ml) 18. Ethanol70% 19. TE Buffer35ul (8ml) 2.2. DNA purification method from tissue: The following is the method used to extract the DNA from the archived sample of breast tissue. Lysis of Formalin-Fixed, Paraffin-Embedded (FFPE) Tissue i.Placed 10-50 mg of 10- to 35- µm thick paraffin sections into an appropriate tube. If using a larger amount of tissue, adjust the reagent volumes accordingly. ii.Diluted 4  µl of â€Å"Proteinase K† into 600  µl of â€Å"Tissue and Cell Lysis Solution† for each sample, and mixed. iii.Added 600  µl of â€Å"Tissue and Cell Lysis Solution† containing the â€Å"Proteinase K† to the sample and mixed. iv.Incubated at 65 °C for 30 minutes; followed by a brief (10 seconds) vortex mix. v.Cooled the samples to 37 °C and added 2  µl of â€Å"RNase A† to the sample; mixed thoroughly. vi.Incubated at 37 °C for 30 minutes. vii.Placed the samples on ice for 3-5 minutes and then preceded with total DNA precipitation (below). Precipitation of Total DNA Added 300  µl of â€Å"MPC Protein Precipitation Reagent† to 600  µl of lysed sample and vortex vigorously for 10 seconds. ix.Pellet the debris by centrifugation at 4 °C for 10 minutes at ?10,000 x g in a microcentrifuge. If the resultant pellet was clear, small, of loose, added an additional 25  µl of â€Å"MPC Protein Precipitation Reagent†, mixed, and pellet the debris again. x.Transferred the supernatant to a clean microcentrifuge tube and discarded the pellet. xi.Added 500  µl of â€Å"isopropanol† to the recovered supernatant. Inverted the tube 30-40 times. xii.Pellet the DNA by centrifugation at 4 °C for 10 minutes in a microcentrifuge. Carefully poured off the â€Å"isopropanol† without dislodging the DNA pellet. Rinsed twice with 70% â€Å"ethanol†, being careful to not dislodge the pellet. Centrifuged briefly if the pellet was dislodged. Removed all of the residual ethanol with a pipet. xv.Resuspended the DNA in 35  µl of â€Å"TE Buffer†. Source: Epicentre Biotechnologies 3. Result: DNA is extracted by a DNA histological processing using PCR and DNA extraction techniques. These are techniques used to extract, amplify and copy small segments of DNA. It is fast and inexpensive because significant amounts of a sample of DNA are necessary for molecular and genetic analyses (Mendizabal et al., 2008). 3.1. DNA Extraction: DNA was extracted by using DNA extraction protocol written in the method section. In the DNA extraction different solutions were used for example Proteinase K enzyme is used to digest protein and to remove protein contamination from DNA and to get to the pure DNA (Ebeling, et al., 1974). Also different machines incubators, vortex and centrifuge were used to break down cell walls. Following the DNA extraction PCR is used to amplify the DNA to find HPV genome. 3.2. PCR: Using the PCR to amplify a segment of DNA firstly the sample is heated so that the DNA denatures or divides into two pieces of single-stranded DNA. After that an enzyme called â€Å"Taq polymerase† synthesizes – builds – two new strands of DNA, using the original strands as templates. This process causes the duplication of the original DNA. Each of the molecules now carries one old and one new strand of DNA. After that each of these strands can be utilized to form two new copies, and the process continues in this manner. More than one billion exact copies of the original DNA segment is achieved by repeating the cycle of denaturing and synthesizing new DNA 35 or 40 times. This whole process of PCR is automated and can be done in just a few hours. A thermocycler machine directs this process and is programmed to change the temperature of the reaction every few minutes to cause DNA denaturing and synthesis. Source: Bruce Fouke’s lab Accessed date: 20/01/11. There are many different types of PCR for example conventional PCR assays using consensus primers and highly sensitive Real-Time PCR (Hedau, et al., 2011). Following are the result of the DNA extracted using a machine called nanoviewer. Many concentration of the DNA extracted are within the good range which is 1.8 – 2.0. This indicates that the samples have not been contaminated with protein. The following table 10 shows the result of the DNA extracted: Breast Tissue sample numberConcentration ng/ulResults 33101.5 ng/ulA260/280 = 1.990 847.5 ng/ulA260/280 = 1.939 7733 ng/ulA260/280 = 1.886 5415.9 ng/ulA260/280 = 1.904 76105.5 ng/ulA260/280 = 1.835 2593.0 ng/ulA260/280 = 1.958 1229.5 ng/ulA260/280 = 1.735 1326.0 ng/ulA260/280 = 1.877 7143.0 ng/ulA260/280 = 2.014 4. Health and Safety: The health and safety procedures were followed according to the requirement of the laboratory for this project and a copy of COSSH assessment was given to the laboratory technical staff and to the project supervisor 5.Ethical Issues: Approval of UK’S ethical committee regarding the usage of the breast tissue samples has already been granted to the project supervisor and hence there is no need of further ethical approval for this project. 6. Literature search: To understand the scope of the HPV and breast cancer very well 15 abstracts have been submitted at the beginning to the project supervisor that were conducted by many journals and research papers. 7. Statistical analysis: This project is a laboratory based and therefore does not require any statistical analysis to be carried out. 8. Treatment: Breast cancer is treated with surgery, radiotherapy, chemotherapy and drugs. There are many drugs that are used to either treat or reduce the risk of breast cancer and following are some example of these drugs: 8.1. Drugs: Table11 below shows the list of drugs used for breast cancer treatment Herceptin (chemical name: Trastuzumab ) Tamofen (chemical name: Tamoxifen) Arimidex (chemical name: anastrozole) Aromasin (chemical name: exemestane) Avastin (chemical name: bevacizumab) Carboplatin (brand name: Paraplatin) Cytoxan (chemical name: cyclophosphamide) Daunorubicin (brand names: Cerubidine, DaunoXome) Doxil (chemical name: doxorubicin) Ellence (chemical name: epirubicin) Thiotepa (brand name: Thioplex) Trelstar (chemical name: triptorelin) Tykerb (chemical name: lapatinib) Vincristine (brand names: Oncovin, Vincasar PES, Vincrex) Xeloda (chemical name: capecitabine) Some of the drugs that are used are explained below. Tamoxifen: is a drug that uses SERMs (selective oestrogen receptor modulator) that attaches to the oestrogen receptors in breast cells and blocks the effects of oestrogen (Lacroix, et al., 2010). Uses: to treat men and both pre-menopausal and post-menopausal women, typically is used to: shrink large, hormone-receptor-positive breast cancers before surgery reduce breast cancer risk in undiagnosed women at higher-than-average risk of developing breast cancer However Tamoxifen is very cost effective (Noah-Vanhoucke, et al., 2011) Side effects: irregular menstrual cycles vaginal discharge or bleeding depression endometrial cancer 8.2. Trastuzumab: is a drug that uses HER2 (human epidermal receptor 2) inhibitors that works against HER2-positive breast cancers by blocking the ability of the cancer cells to receive chemical signals that tell the cells to grow. Uses: treat metastatic, HER2-positive breast cancer (Barok, et al., 2011) shrink large, advanced-stage, HER2-positive cancers before surgery Side effects: diarrhea anemia abdominal pain 9. Cancer prevention: 9.1. HPV Vaccines and Cervical Cancer: Cervical cancer and sexual behaviour of population are directly proportional to each other, recent study shows that in the U.K HPV prevalence and possession increased consistently with increasing numbers of lifetime sexual partners, regular partners, and new partners in the last 5 years (Almonte, 2011). The two prophylactic vaccines Cervarix and Gardasil consist of virus-like particles (VLPS), these are recombinant viral capsids made by expressing HPV 16 and 18 L1 proteins in insect cells through the baculovirous (cervarix) or HPV 6, 11, 16 and 18 L1 proteins in yeast cells (Gardasil) (Kahn 2005; Wang 2007; Kirnbauer et al., 1993). The virus-like particles (VLPS) contains no viral DNA and therefore would not in any case cause an infection or cervical cancer in recipients (Wang, 2007). Cervarix: GlaxoSmithKline (GSK) produces cervarix vaccine; it is a bivalent containing HPV 16 and 18 L1 virus-like particle vaccines that works against HPV 16 and 18 infections and cervical cancer (Bayas et al. 2008; Govan 2008). A phase II study illustrated that Cervarix was 91.6% efficacious against occasional infections and 100% effective against persistent infection (Harper et al. 2004). Cervarix is made up of an ASO4 adjuvant which contains aluminium hydroxide and 3-O-deacylated monophosporyl lipid (MPL), ASO4 helps improve the immune system (Bayas et al., 2008). Gardasil: Gardasil is developed by Merck and Co; it is a quadrivalent vaccine containing HPV 6, 11, 16 and 18 virus-like particles (Adams, et al., 2007; FDA, 2006). A phase II efficacy study of Gardasil results demonstrated that the vaccine has 90% efficacy in preventing incident HPV infection and cervaical cancer (Viller, et al., 2005). In June 2006 Gardasil was licensed by the FDA for use in young and adult females between the ages of 9 to 26 for the prevention of cervical cancer, genital warts and precancerous lesions (FDA, 2006), it was also approved in September 2008 for the prevention of vaginal and vulvar cancers caused by HPV 16 and 18 (FDA, 2008). Both of the above vaccines are given in a series of three 0.5ml immunisations over a time period of six months prior to a young person becomes sexually active (Long III, et al., 2007; WHO, 2007). Figure 24 shows how the HPV DNA is detected in cervical cancer. The above figure adopted from: Global Link (2008) Date accessed 07/04/11 9.2. Breast Cancer Vaccine: Vaccine has been developed firstly against cervical cancer and now the scientists are trying to develop a vaccine against breast cancer, however scientists are trying to develop a vaccine which could be useful against all the different strains of HPV such as 16, 18, 33 worldwide (Armstrong, 2010). Prognosis: Table 14 shows the five year survival rates for colorectal and breast cancer. (Howlader, et al., 2011). There is only 23% survival rate for distant spread in breast cancer this shows that there is a need for more research to develop a vaccine against different strains of breast cancers and to treat these cancers affectively and avoid so many deaths. 10. Discussion: Breast cancer is a malignant tumour that starts from cells of the breast. Cancer occurs due to mutations in the genes responsible for controlling the growth of cells thus cells are unable to function properly (Sariego, 2010). Human papillomavirus (HPV) is considered as an aetiological agent for many cancers such as cervical cancer, breast cancer etc. High risk HPV types causes cancer by integrating into the host genome and causes disruption and loss of some of the viral genes such as L1 and L2 genes and also increases the expression of the early genes (Wang, 2007; Mera, 1997). The aim of the research was to find out the association of HPV with breast cancer involving DNA extracted from archived breast tissue samples using DNA extraction method. This DNA sample could be amplified using PCR to find HPV genome specifically targeting E1 gene. This is conjunction with other studies in which samples were amplified using consensus primers CpI CpIIG and targeted the E1 gene in a region conserved for 99% of most common HPV subtypes (Mendizabal, et al., 2008). Given that the tissue samples were not fresh but were paraffin embedded which are not as good as fresh tissue samples because formalin fixation could denature the tissue during sectioning and also the DNA extracted from FFPE (formalin fixed paraffin embedded) tissues are usually at low concentration and disjointed (Shi, et al., 2006). Additionally the experiment was carried out very successfully because most of the results that have been obtained were between the ranges of 1.8 to 2.0, which are regarded as pure DNA sample and therefore contains no protein contamination. A positive and negative control should be carried out while amplifying the DNA because a positive control makes sure the technique is working satisfactorily by using a reacting material relatively similar to the test material and negative control tests the specificity of the reaction and ensures there are no false positives (Mendizabal, et al., 2008). Although good results have been achieved however there were some variations in the purity of DNA extracted from the breast tissue samples and that depends on many different factors such as some tissue samples were darker in colour than normal which suggests the samples were not as fresh therefore it gave a lower result than the normal range of 1.8 to 2.0. Additionally the low results also depended on the way the whole experiment was carried out, there had been some mistakes in adding or mixing different solutions and mistakes were constantly recorded in the lab book therefore the same mistakes were not repeated again. Moreover many different techniques have been learnt from this project including the usage of centrifuge, vortex, incubator and nanoviewer. Carrying out this project has provided a full understanding on how to engage in the practical work which is beneficial in future laboratory projects, this is an ongoing work by the supervisors of the project to try and identify the association between the HPV and the breast cancer, if successful then a broader vaccine could be developed against all different strains of HPVs such as HPV16, 18 worldwide and to cure not only breast cancer but also many different types of cancers such as, cervical cancer, head and neck Squamous cell carcinoma, genital warts etc. This will reduce the amount of vaccination given to each patient and also it will have tremendous effect on the quality of life and will solve many problems and save many lives. Furthermore many studies have been carried out to find out the presence of HPV in breast tissue. Some were successful by getting 86.21% positivity of HPV infection in breast cancer (de Villers et al., 2005) this was in conjunction with other studies that have been successful in obtaining high positive result (Hening, et al., 1999; Gumus, et al., 2006; Kan, et al., 2006, Li, et al., 2002). Additionally according to a largest investigation on breast carcinoma specifically analysing mammoplasty and fibroadenoma specimens as a control group the authors were able to detect HPV DNA in 24.5% of the breast carcinomas but were unable to detect any in benign breast specimens (Damin, et al., 2004). However other authors have either achieved low positivity (Kroupis, et al., 2006; Tsai, et al., 2005) or HPV was totally absent ( Lindel, et al., 2007; Gopalkrishna, et al., 1996). Therefore there are two different views on the association of HPV with breast cancer as it has been indicated by the above studies, which is normal because scientists can have different opinions sometimes. This project was limited because only the DNA has been extracted and the DNA was not amplified by using PCR, in the future if this project were to be continued PCR can be used to amplify the gene from the DNA extracted in this project and the investigation can be expanded and more information can be obtained. References: Adams, S., Greeder, L., Reich, E., Shao, Y., Fosina, D., Hanson, N., Tassello, J., Singh, B., Spagnoli, G. C., Demaria, S., Jungbluth, A. A. (2011) Expression of cancer testis antigens in human BRCA-associated breast cancers: potential targets for immunopreventionCancer Immunol Immunother 262 (11) 1005-1007. Akil N, Yasmeen A, Kassab A, Ghabreau L, Darnel AD, Al Moustafa AE. (2008) High risk human papillomavirus infections in breast cancer in Syrian women and their association with Id-1 expression; a tissue microarray study. Br J Cancer. 2008, 99(3), 404-7. Almeida, C. A., Barry, S. A. 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The Significance Of The Cross Essays - Christology,

The Significance of the Cross The significance of the cross has changed over the years. In the beginning before Christ gave himself up for our sins the cross was viewed as a sign of pain and suffrage. The Romans used to execute criminals and enemies of the state on them. The were hung along the road showing the sarrows of people. It was the most shameful way to die. The pain and anguish it caused was unbearable and feared by all. Except Roman citizens. The pain and shame of death on the cross was so bad that the Romans would not inflict it on their citizens. In their society it was like the electric chair is today. Except for the fact that we inflict that pain on any human being. The Romans new of the pain that it caused human beings but yet they still inflicted it on people. The cross was a sign of the cruelty of the Roman Empire. It showed what they believed in as a society. Roman was not a society of pity, forgives, or any other act of caring towards human beings that were not Romans. The cross showed their society as that of almost barbaric tribes. The cross and the gladiatorial coliseums were the same thing, ways to punish and kill their enemies. The Romans used this way of execution for Christ. They put him up onto the cross and changed its whole meaning for millions of people. With that one act the cross lost most of its spur to someone's dignity for a Christian. Because of that act the cross has become a sign of salvation. The followers of Christ see the cross as their redemption and their only way of entering the Kingdom of God, Heaven. The cross is a symbol that people use to remind themselves that there is a reason to go on. The cross means they can now enter heaven because Jesus died for them one of the most brutal ways ever. The cross now is a reason for people to live the life that Christ lived. It gives them hope and joy every time they look at. The cross is still a sign of pain to people, but that pain is greatly overshadowed by the love for them that is shown on it. Everyone who looks upon the cross cannot help but feel the pain Jesus went through for their souls to have the ability to be joined with him in Gods greatest gift to man, Heaven. The anguish inflicted on the cross does not equal what the act gives to everyone who tries to live his life as best he can. The cross was once a sign of the ultimate shame. It was pain in its purest form, but now that great pain has been overshadow by the sacrifice born of love. The ultimate show of love has made the cross what I believe it is today, the only sign that shows the ability of salvation and a life in heaven.