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Dissertation presentation

Transcript: Barley Stripe Mosaic Virus (BSMV) has an unusual tripartite genome Phytoene, carotenoid pathway and phytoene desaturase (PDS) genes Susceptible to photo-bleaching Agrobacterium-carrier of BSMV used for secondary infection via Nicotiana benthamiana Infected N. benthamiana leaves are used to produce a sap, which is then used to infect the wheat plants Methodology Adapted from Yuan et al., 2011 I am incredibly grateful for the assistance, laboratory equipment (including the PDS-gene sequence) and support from all of those working in Dr. Sadanandom’s group; special thanks must go to Jack Lee, for all his help, guidance and support. Experiment components Figure 5 – Time-course sample ‘g’; Left to right: M, EV, PDS with actin and then PDS primers Stage 3: Time course and real-time PCR Samples taken alternate days until 14 DPI There was no evidence that the average leaf length for the PDS-treated wheat plants differed from that of the ‘mock’ plants Conclusions Introduction Conclusions Figure 7 – real-time PCR time-course output showing relative levels of PDS expression in wheat plants at each time point Time-course: quantitative initial increase in transcript levels of the PDS gene, which by 10-12 DPI were noticeably down-regulated Secondary infection allows systemic viral infection to develop in the wheat plants Pre-inoculation application of carborundum powder and sap inoculation using the intermediate host – adapted methodology is capable of producing fast, effective and efficient results for PDS gene silencing VIGS has helped to identify novel pathways involved in defence and pathogenesis, investigate biosynthetic pathways, and characterise stress or hormone response gene. The use of VIGS to investigate disease and pathogen resistance could be particularly important – crop improvement? Figure 3 – Gel electrophoresis, from left to right: hyperladder, E2-5, M2-5 & P2-5 (actin primers) A marker of Barley Stripe Mosaic Virus-Induced Gene Silencing in wheat Stage 2: Growth indicators and viral infection Monitored growth indicators for the wheat plants – leaf length, number of leaves, and any signs of the PDS phenotype There is no difference between the average leaf length of PDS­-treated and the mock-treated wheat plants Infection with BSMV does not affect plant growth Thank you for listening Any questions? Quantification of RNA silencing: Comparing RNA transcript levels of common, usually-transcribed genes, such as actin or ubiquitin, to the PDS gene Our central hypothesis: after inoculation, RNA transcript levels of the PDS gene in infected plants will be initially increased, and subsequently down-regulated, when compared to actin or ubiquitin. Establish the effectiveness of VIGS and help to confirm the use of BSMV as a suitable VIGS vector in wheat: Confirm that any phenotypic changes are due to silencing of the target gene, rather than symptoms of BSMV infection. Certain plant growth indicators (i.e. total number of leaves and total leaf length) were measured Secondary hypothesis: growth indicators will not be affected by BSMV infection in any of the treatments Primary aim of the experiment: To establish a workable mechanism for Virus Induced Gene Silencing in wheat Stage 1: Preliminary investigation RNA extraction and cDNA synthesis techniques were workable Actin primers were used to compare RNA levels of PDS in semi-quantitative PCR and gel electrophoresis – successfully visualised gene expression in different ages of wheat plants Stage 3: Time course and real-time PCR Showed an initial increase in PDS-gene expression then a noticeable and marked decline in PDS-gene expression All the wheat plants inoculated with the PDS-insert Agrobacterium showed the PDS phenotype in at least one leaf RNA transcript levels of the PDS gene were clearly affected, increasing within 2 days of infection, and then steadily decreasing Victoria Zoë Harris Grey College – Biology (Single Honours) – 2013 Supervisor: Dr. Ari Sadanandom Figure 8 – ‘white leaf phenotype’ in infected wheat leaves (21 DPI) Use BSMV constructs to silencing two genes simultaneously? PDS could still be an important marker – combining the BSMV  and  genomes could increase the speed of viral spread within the plant as well as increased stability of the silencing. Quantitative PCR techniques are far better potential evidence than the more qualitative nature of phenotypic decolouration Using the PDS gene alongside another gene in simultaneous silencing could work well as a marker for (simultaneous) gene silencing The use of VIGS does have clear limitations VIGS can be transient and incomplete; the vector can be inherently unstable; and efficient infection of the host plant, resulting in appearance of the established silenced phenotype, frequently occurs in a minority of the plants originally infected Use of secondary infection via an intermediate host - Improves the efficacy of wheat infection? Results

Dissertation Presentation

Transcript: How Far is Jesus of Nazareth's Atoning Sacrifice a Retelling of the Aqedah? An Analysis of the Texts including and surrounding Genesis 22:1-19 and John 19:16-37 Spark! Explore The HB concepts of child sacrifice and compare to NT Isaac and Jesus- sacrifice of the 'only/beloved son Atonement Child sacrifice outside of the Bible. A focus on Gen.22 and Jn.19 Books Journal Articles Internet Articles The Bible! Topics include human sacrifice, atoning sacrifice, Greek mythology and the beloved son motif. Commentaries on Genesis 22:19, Epistle to the Hebrews (esp. ch.10) and John 19. Child Sacrifice in the HB What is sacrifice? Genesis 22:1-19 Molek Laws forbidding/requesting child sacrifice Evidence outside of the Hebrew Bible 2. Isaac said to Abraham [his father, 'Behold there is the fire and the wood, but where is the lamb] 3. for the burnt-offering?' And Abraham said to [Isaac, his son, 'God will provide the lamb] 4. for himself.' Isaac said to his father 'T[ie me well’] 5. The angels of holiness were standing weeping above [the alter] 6. his sons from the earth. And the angels of M[astemah] 7. were rejoicing and saying, 'Now he (Isaac) will be destroyed’. And [we shall see] whether 8. he will be found weak and whether A[braham] will be found unfaithful [to God.'...” Jesus' Atoning Sacrifice Atoning sacrifice Exploration of the Epistle to the Hebrews How does this relate to the Aqedah? Passion Narrative (Jn.19) Similarities with the Aqedah Similarities with Passover narrative (Exod.12). amnos Lamb sacrificed as a sin offering The paschal lamb The lamb of Isaiah Conclusions Ultimately, the NT is a retelling of the HB. The Aqedah as etiology New covenant Questions? "Consecrate to me every first born male. The first offspring of every womb among the Israelites belongs to me, whether human or animal" (Exod. 13:2). Comparison to Jesus' Atoning Sacrifice "I will set my face against him and his family and will cut them off from their people together with all who follow him in prostituting themselves to Molek" (Lev.20:5) Or- "sacrifice is the removal of obstacles that threaten the relationship between God and God's people" “A jar of vinegar wine was there, so they soaked a sponge in it, put the sponge on a stalk of the hyssop plant, and lifted it to Jesus’ lips” (Jn. 19:29) Exegesis of Gen.22:1-19 Research The relief at Pozo Moro Graves at Carthage DSS Jubilees 17 Against Sacrifice hyssōpō (on hyssop) or hyssō (on a javelin) Ram Only Son Knife Wood Mountain Child Sacrifice vs. 'mlk' sacrifice Conclusions Research “take a bunch of hyssop, dip it into the blood in the basin and put some of the blood on the top and on both sides of the door frame” (Exod. 12:22) How does the Epistle represent atoning sacrifice? "He too shared in their humanity so that by his death he might destroy him who holds the power of death...and that he might make atonement for the sins of the people" (Heb.2:14-17). Scholarly opinion "They have built shrines to Baal- to put their children in the fire as burnt offerings to Baal- which I never commanded, never decreed, and which never came to my mind" (Jer.19:5). Eissfeldt “By faith Abraham, when put to the test, offered up Isaac. He who had embraced the promises was ready to offer up his only son, of whom he had been told, “It is through Isaac that your descendants will be reckoned.” Accounting that God would be able to raise him up, even from the dead, whence he received him also in parable.” (Heb.11:17-19). Child Sacrifice in the HB Stavrakopoulou For Day of Atonement The Gospel of John- The Passion Narrative (Jn.19) How does this relate to the Aqedah? Epistle to the Hebrews Is there evidence of atoning sacrifice in the NT? Francesca Stavrakopoulou IDB- "slayings take place with the fundamental objection of engendering changes at a supra-human level" "Any Israelite or any foreigner residing in Israel who sacrifices any of his children to Molek is to be put to death" (Lev. 20:2) "The topheth has long been ready for him. He too is ready for Melech" (Is.30:33) 4Q225 Quotes from the Bible Spark Basis of understanding A look at King Manasseh and Child Sacrifice A focus on archaeology "You must give me the first born of your sons" (Exod. 22:29). Atonement "I, in turn, gave them laws which were not good, and rules by which they could not live: When they set aside every first issue of the womb, I defiled them by their very gifts" (Ezek.20:25).

Dissertation Presentation

Transcript: Malawi - Society and Culture Literature Review Aims and Objectives Research Questions Quantitative Data An Exploratory Study of the Childbirth and Parenting Experiences of Women with Mental Disorders in Malawi Qualitative Data “R: Some of the drugs makes one sleep therefore you rarely concentrate on the children since you are most of the times asleep, everything seems to keep you tired.” -RM04, diagnosed in 1991 with severe depression, she carers for 2 extra children along with her own 2, student at the polytechnic “R: It is all same blood it is the same way you stay a grandmother or a sister would take your child, so yes I did give them the go ahead they are like hers as well.” -EG09, this participant has BPAD, has never been to school, she is divorced with 4 children, but has a very supportive immediate family. Figure 3: QECH Introduction Methods Key highlights of the Results Discussion Personal Reflection Conclusion References 8th May 2012 Figure 4: ZMH *Participant does not know her own age, but her notes say 25+, so this is an estimate. Summary and Conclusion Discussion Source: “R: It upsets me because I feel I have failed my responsibility as a mother.” -JN02, participants with severe depression, and HIV, she is remarried, and both children current live with her neighbour. “R: I was stressed... I was thinking on how I was going to raise the children all by myself.” -AM12, this participant is also HIV positive, is widowed and has 4 children, who are living in a government orphanage Methods Search strategy Study Design Data Required Study Population Sampling and Sample Size Data Collection Procedure Data Handling Data Analysis Piloting “R: At first I never used to understand, but I think it is God’s secret where this happens when he wants to communicate to someone, so I take it as a way of communication and to reflect on one’s life” -JN02, participants with severe depression, and HIV, she is remarried, and both children current live with her neighbour. Figure 2: Human Developement Index, Malawi United Nations Development Programme. International Human Development Indicators. Source: (accessed 21/04/12) Research Highlights Belle Boyle 1) Food and Agriculture Organisation. Nutritional Country Profiles. (accessed 23/04/12) 2) WHO, Maternal mental health and child health and development in low and middle income countries. 2008, World Health Organisation: Geneva. 3) Sawyer, A., S. Ayers, and H. Smith, Pre- and postnatal psychological wellbeing in Africa: a systematic review. Journal of Affective Disorders, 2010. 123(1-3): p. 17-29. 4) Collaboration for Evidence Based Health Care in Africa. Welcome to CEBHA. (accessed 12/03/12) 5) Silverman D. Qualitative research. 3rd edition. London: Sage Publishing; 2011. Page 131. 6) Annells M. Grounded Theory Method: Philosophical Perspectives, Paradigm of Inquiry, and Postmodernism. Qual Health Res. 1996:6. 379-393. 7) Creswell JW, Hanson WE, Clark Plano VL, Morales A. Qualitative Research Designs: Sel33ection and Implementation. The Counseling Psychologist [Internet]. 2007 Mar 1 [cited 2012 Feb 29];35(2):236–64. Available from: 8) 41) Pope C. Qualitative research in health care: Analysing qualitative data. BMJ [Internet]. 2000 Jan 8 [cited 2012 Mar 6];320(7227):114–6. Available from: 9) Patel V. Mental health in low- and middle-income countries. British medical bulletin [Internet]. 2007 Jan 1 [cited 2012 Mar 5];81-82(1):81–96. Available from: “R: my mother does not want me around, so do my relatives, it is only my father I am in good relation with, with my children except the first born who runs when he sees me.” -EK11, 26 year old with 3 children, has also had 3 miscarriages, despite these problems she still lives with her mother. “R: He just said we are separated. And I know it was because of my condition because he just left for work and never returned.” -MW15, participant is diagnoses with schizophrenia, is divorced, and children live with her grandmother. Thank you for your time. Any Questions? Personal Reflections Introduction and Background Figure 1: Map of Malawi Source: †Participant referred to one admission during the interview, but could have been admitted more times. ‡ Participant’s date of initial diagnosis is an estimate based on her year of school which she was in as she cannot remember her age when she was diagnosed. ¥ This participant was initially diagnosed in November 2011, however she suffered her first episode of illness in 2001, but at this time consulted a traditional healer only. Discuss and Interpret the key Findings Strengths and Limitations Implications and Recommendations Presentation Summary

Dissertation Presentation

Transcript: The decline of geography in Bolton's primary schools and the role politics has in the decline. "64% of teaching was either satisfactory, unsatisfactory or poor, whilst only 36% was good. very good or excellent." (Catling et al, 2003) "in approximately one in ten primary schools visited , geography was more or less disappearing." (OFSTED, 2011) "More than 100 secondary schools do not enter a single pupil for a GCSE exam in the subject" (The Independent, 2011) "only some 50% of primary school teachers feel confident teaching geography." (Rawlinson, 2003) my observations are at risk of my bias "Both practical and classroom" "not seen as a main subject by other teachers." Observation Joseph McQuaid Qualitative St John's Q) Do you enjoy doing geography? A) "We don't really do geography." Study Area What are children's perceptions of geography in their schools? Do they enjoy it? Geography is still there, but only through the use of 'topics'. 'Topics' include geography, history, art and others !? "Closure of several University geography departments, the decline in number of geography students wishing to enter teaching and a diminishing of the subjects status in English schools" (Rawlinson, 2003) Thank You Geography is no longer seen as a main subject in the National Curriculum in Bede Burn Primary school (cc) photo by jimmyharris on Flickr teachers may not be totally honest Students feel like they do not study geography, and teachers feel they are teaching geography when teaching the topic (geography of the olympics). St Teresa's St John's the Evangelist St Columba's OFSTED, (2011), Geography declining in schools, NR2011-04, Manchester [OFSTED]. OFSTED, (2008), Geography in schools – changing practice, 070044, Manchester [OFSTED]. OFSTED, (2011), Report Summary, Geography, learning to make a world of difference, 090224, Manchester [OFSTED]. political side of the research government spending on education and OFSTED reports new curriculum? qualitative St Columba's Teacher's lessons are "not good enough in half of schools." (Telegraph, 2011) Adey, K., Biddulph, M., (2001) 'The Influence of Pupil Perceptions on Subject Choice at 14+ in Geography and History', Educational Studies, 27: 4, 439 — 450 BBC, (2011), Geography declining in many English schools – OFSTED [Online], available: [27/02/2011]. Boyle, Bill and Bragg, Joanna(2006) 'A curriculum without foundation', British Educational Research Journal, 32: 4, 569 — 582 Butt, Graham , Weeden, Paul , Chubb, Steven and Srokosz, Anne(2006) 'The State of Geography Education in English Secondary Schools: An Insight into Practice and Performance in Assessment', International Research in Geographical and Environmental Education, 15: 2, 134 — 148 Castree, N., Fuller, D., Lambert, D., (2007), ‘Geography without borders’, Boundary crossings, 31, 129 – 132. No specialism (cc) photo by Metro Centric on Flickr limitations Indepth interviews Students not even sure what geography is!? Asking individual children at Bede Burn: Brainstorm research questions "I'm trying to bring geography into my class everyday." Quantitative & Qualitative (cc) photo by Metro Centric on Flickr Observations Q) What do you learn in geography? A) "What, like places? We're doing about pharaohs in Egypt at the moment." References Geography and History taught together Taught as topic: Egypt, Olympics Not aware of geography lesson If Children don't realise they are doing Geography they will never develop a love for the subject. problem? Questions to pupils every teacher in each school = 6 teachers per school Conclusion Government are very irrational methodology Results Rawlinson, S., Essex-Cater, L., Bolden, D., and Constable, H., (2003) 'Have Geographers Lost Their Way? Issues relating to the recruitment of geographers into school teaching', Journal of Geography in Higher Education, 27: 1, 39 — 56 Rose, J., (2009), ‘Independent review of the Primary Curriculum: Final Report’. Stannard, K., (2003), ‘Earth to academia: on the need to reconnect university and school geography’, Area, 35.3, 316 – 322. The Telegraph, (2011), Geography lessons ‘not good enough in half of schools’ [Online], available: [27/02/2011]. The Telegraph, (2011), without geography, the world would be a mystery to us [online], available: [27/02/2011]. literature review St Teresa's Primary Geography? So far ... Is the decline of geography significant in primary schools today? Q) Top 5 subjects? A) "Literacy, Science, PE, Art, Maths." Rachael Locker's bum stinks (cc) photo by Franco Folini on Flickr Ask students how they feel about geography in their schools using sticky notes to involve students words or pictures Geography graduate teacher Interview with geography graduate teacher, St

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