No description available.

Retroelements are genomic parasites which make up ~42% of the human genome and 38% of the mouse genome. Most are degenerate, but a large number have relatively intact promoter elements, suggesting that they are capable of transcription. Transcriptionally active retroelements can perturb normal transcription units in their vicinity through a variety of mechanisms, leading to phenotypic effects and in some cases disease. This phenomenon of transcriptional interference has been observed in organisms as diverse as maize, Drosophila, and the mouse. We analysed the extent of retroelement transcription in normal and diseased tissues, by searching the mouse and human EST databases for transcripts originating in retroelement promoters, and found a large number of transcripts from LINEs, SINEs and ERVs. Retroelement transcripts were found to be initiated in both sense and antisense orientations, and to be equally as common in normal and diseased tissue. Several of these transcripts were chimeric, appearing to initiate in retroelements and reading through to cellular genes, suggestive of transcriptional interference. We have used transposon display to identify and recover retroelement transcripts in the mouse. Transcripts initiated in LINE, SINE and ERV promoters are numerous, and many are chimeric with cellular genes. Although the numbers of recovered chimeric transcripts are too large to permit rigorous analysis of more than a small proportion, some of those we have studied further appear to be authentic transcripts that may represent interference with the canonical promoters of the genes in question. Our results suggest that transcriptional interference by retroelements may be a relatively common occurrence in mammals.

Master of Science / Statistics / Nora M. Bello / Big bluestem (Andropogon gerardii) is a wide-ranging dominant prairie grass of ecological and agricultural importance to the US Midwest while edaphic subspecies sand bluestem (A. gerardii ssp. Hallii) grows exclusively on sand dunes. Sand bluestem exhibits phenotypic divergence related to epicuticular properties and enhanced drought tolerance relative to big bluestem. Understanding the mechanisms underlying differential drought tolerance is relevant in the face of climate change. For bluestem subspecies, presence or absence of these phenotypes may be associated with RNA transcripts characterized by low number of read counts. So called low-count transcripts pose particular inferential challenges and are thus usually filtered out at early steps of data management protocols and ignored for analyses. In this study, we use a plasmode-based approach to assess the relative performance of alternative inferential strategies on RNA-seq transcripts, with special emphasis on low-count transcripts as motivated by differential bluestem phenotypes. Our dataset consists of RNA-seq read counts for 25,582 transcripts (60% of which are classified as low-count) collected from leaf tissue of 4 individual plants of big bluestem and 4 of sand bluestem. We also compare alternative ad-hoc data filtering techniques commonly used in RNA-seq pipelines and assess the performance of recently developed statistical methods for differential expression (DE) analysis, namely DESeq2 and edgeR robust. These methods attempt to overcome the inherently noisy behavior of low-count transcripts by either shrinkage or differential weighting of observations, respectively. Our results indicate that proper specification of DE methods can remove the need for ad- hoc data filtering at arbitrary expression threshold, thus allowing for inference on low-count transcripts. Practical recommendations for inference are provided when low-count RNA-seq transcripts are of interest, as is the case in the comparison of subspecies of bluestem grasses. Insights from this study may also be relevant to other applications also focused on transcripts of low expression levels.

No description available.

Submitted by MARINA MORTATI DIAS null (marina.mortati@gmail.com) on 2016-08-07T23:35:50Z No. of bitstreams: 1 Tese_Marina Mortati Dias.pdf: 1708247 bytes, checksum: 534b59570229fd27eb89d883f8e64df7 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-08-10T13:49:51Z (GMT) No. of bitstreams: 1 dias_mm_dr_jabo.pdf: 1708247 bytes, checksum: 534b59570229fd27eb89d883f8e64df7 (MD5) / Made available in DSpace on 2016-08-10T13:49:51Z (GMT). No. of bitstreams: 1 dias_mm_dr_jabo.pdf: 1708247 bytes, checksum: 534b59570229fd27eb89d883f8e64df7 (MD5) Previous issue date: 2016-07-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Características reprodutivas, como a idade à puberdade, são economicamente relevantes para sistemas de produção de carne. A ocorrência de prenhez precoce aos 16 meses (OPP) é considerada um indicador da idade à puberdade e pode ser utilizada como critério de seleção. A busca por mutações potencialmente causais em genes candidatos pode ajudar a melhorar a acurácia de predição genômica se forem inseridas em painéis de marcadores genéticos. O objetivo desse estudo foi identificar polimorfismos potencialmente causais em genes candidatos associados a características reprodutivas em novilhas, por meio de dois ensaios. No primeiro, foram sequenciadas regiões dos genes PPP3CA e FABP4 em 380 amostras de DNA provenientes de novilhas Nelore, por meio de equipamento MiSeq® (Illumina, Inc). Nestas sequências foram detectadas duas mutações na região do gene PPP3CA e 13 no gene FABP4. Dentre elas, 14 eram SNP e uma era deleção, sendo esta última homozigota em todas as amostras sequenciadas, o que nos leva a acreditar que possa ser uma mutação fixada na raça Nelore. Um haplótipo constituído por quatro SNP no gene FABP4 não teve efeito significativo ao nível de p<0,05 sobre a característica OPP. Todos os SNP que passaram pelo controle de qualidade também foram analisados, porém nenhum teve efeito significativo sobre a característica OPP (p>0,05). No segundo ensaio, foram analisadas sequências de amostras de RNA de novilhas Brangus com o objetivo de identificar polimorfismos nas regiões de 62 genes candidatos associados a características reprodutivas. Detectou-se 1157 SNP nas regiões de 46 daqueles genes. Esses SNP foram comparados com SNP detectados a partir de sequências de amostras de RNA de outras quatro raças (Angus, Brahman, Nelore e Holandês). Cento e setenta e dois SNP foram coincidentes em todas as raças. A partir de sequências de RNA de Brangus também foram detectados 160 novos transcritos na região de 42 dos genes candidatos e cinco genes não anotados que sobrepõem a região de genes candidatos. A detecção de novos transcritos e genes é fundamental para enriquecer a anotação do genoma bovino. As mutações detectadas podem ser utilizadas para a customização de painéis de marcadores, e aplicações de seleção genômica. / Fertility traits such as age at puberty are economically important for meat production system. Occurrence of early pregnancy (OPP) is considered a good indicator of age at puberty and can be used as selection criteria. The search for causal mutations in candidate genes can helpful to improve the accuracy of genomic prediction if used to design low-density chips. The objective of this study was to identify polymorphism in candidate genes associated with puberty in heifers. DNA sequences were obtained from 380 Nellore heifers from exons sequencing of PPP3CA and FABP4 genes through MiSeq® equipment (Illumina, Inc.). From these sequences were found two SNP in the region of PPP3CA gene and 13 variations in the gene FABP4. Among these 15 variations 12 were SNP and one was a deletion, and this deletion was detected in all samples sequenced, which leads us to conclude that this may be a variation between Nellore and Hereford. One haplotype consisting of four SNP located in the FABP4 and nine SNP, that were analyzed separately, had not a significant effect at the p <0.05 on OPP characteristic. Also were analyzed RNA sequences from Brangus heifers in 62 candidate genes associated with puberty with the aim to discovery SNP in these regions. Were detected 1157 SNP in the region of 46 gene. These SNP were compared with SNP detected from RNA sequences from four breeds, Angus, Brahma, Nellore and Holstein. One hundred and seventy-two SNP were matched in all breeds. From Brangus RNA sequences were also detected 160 new transcripts in the region of 42 candidate genes and five genes not annotated that overlap the region of candidate genes. Detection of new transcripts and genes is essential to enrich the bovine genome annotation. The mutations detected can be used for the customization of low density chips and future genomics selection strategies.

No description available.

No / Metastatic melanoma is the leading cause of skin‑cancer related deaths and while in recent years some progress has been made with targeted therapies, there remains an urgent unmet need for novel therapeutic treatments and reliable diagnostic, prognostic and predictive biomarkers. The emergence of next generation sequencing (NGS) has seen a growing appreciation for the role played by non‑coding genomic transcripts in regulating gene expression and by extension impacting on disease progression. The long non‑coding RNAs (lncRNAs) represent the most enigmatic of these new regulatory molecules. Our understanding of how lncRNAs regulate biological functions and their importance to disease aetiology, while still limited, is rapidly improving, in particular with regards to their role in cancer. Herein we review the identification of several lncRNAs shown to impact on melanoma disease progression and discuss how these molecules are operating at the molecular level.

Facioscapulohumeral muscular dystrophy(FSHD) is a disorder characterized by muscle weakness and wasting (atrophy). This disease is typically inherited as autosomal dominant and has a complex genetic and epigenetic etiology. Our collaborator had differentiated healthy human pluripotent stem cells(iPSC) into skeletal muscles and exploited ISO-Seq to explore cell gene expression and transcript alternative splicing usage profile during 8 differentiation stages. Later, stage specific gene differential expression, transcript alternative splicing, gene ontology and novel gene/transcript were analysed to characterize the feature of each stage during the differentiation. In terms of expressed genes with more than or equal to 5 transcripts, each stage had shown their own stage specific features. About transcripts, iPS, S1, ADM.D0, ADM.D4 have about 30% to 40% more total transcripts than the rest 4 stages. 4 kinds of alternative splicing events are generally distributed and S2 stage has the least alternative splicing events potentially due to technical reasons. As for gene differential expressions, ADM.D4 has considerable amount of differential expressed genes with 5 other stages and it has minor difference with ISM.D4 and S3 stages(they are all myotubes cells). The gene ontology analysis is performed according to the results of previous step, stage specific GO terms are revealed.

Facioscapulohumeral muscular dystrophy(FSHD) is a disorder characterized by muscle weakness and wasting (atrophy). This disease is typically inherited as autosomal dominant and has a complex genetic and epigenetic etiology. Our collaborator had differentiated healthy human pluripotent stem cells(iPSC) into skeletal muscles and exploited ISO-Seq to explore cell gene expression and transcript alternative splicing usage profile during 8 differentiation stages. Later, stage specific gene differential expression, transcript alternative splicing, gene ontology and novel gene/transcript were analysed to characterize the feature of each stage during the differentiation. In terms of expressed genes with more than or equal to 5 transcripts, each stage had shown their own stage specific features. About transcripts, iPS, S1, ADM.D0, ADM.D4 have about 30% to 40% more total transcripts than the rest 4 stages. 4 kinds of alternative splicing events are generally distributed and S2 stage has the least alternative splicing events potentially due to technical reasons. As for gene differential expressions, ADM.D4 has considerable amount of differential expressed genes with 5 other stages and it has minor difference with ISM.D4 and S3 stages(they are all myotubes cells). The gene ontology analysis is performed according to the results of previous step, stage specific GO terms are revealed.

Abstracts THE TAINT This is a memoir of Ray “Poss” Ide, a man who has carried the taint of a horrifying crime since he was seventeen years old─a crime he claims he did not commit. The crime was the rape of a fourteen-year-old girl. The Taint looks at the time leading up to his conviction for rape, including the years he spent in Westbrook Boys’ Reformatory and Boggo Road Gaol as a minor, and then his sentence served in NSW gaols with such inmates as Stephen Bradley, the man who kidnapped and murdered Graham Thorne, and the bizarre but rather likeable Dave Scanlan, known for his exploits as “the Kingsgrove Slasher”. In prison, Scanlan encouraged him to become an elite sportsman and released, Poss was recruited by Canterbury Eels football team; but just on the point of making a name for himself in Sydney, someone discovered that he had been gaoled for rape, and humiliated, he left the team. He moved back home to Queensland via Grafton, NSW (where he met and later married the Jacaranda Queen) and continued his sporting career, but never again in the Big League. He became the Manager of the Waterside Workers’ Club and helped prevent a turf war between the Wharfies and members of the Painters and Dockers. Poss is now working with lawyer Robert Bax to have his case re-opened. He believes his story to be a cautionary tale for young men. It is a chronicle of social change, including the sexual revolution of the sixties, the confrontations with “the demons” in the streets of Brisbane during the Joh era of Bible-bashing fundamentalism and police corruption. It’s a story about what really went on behind closed doors in institutions where vulnerable children were preyed upon, in gaols where men were expected to become beasts. It is a story of how the taint of a terrible crime affected a man’s whole life. “QUESTIONS OF OWNERSHIP” Writing someone else’s story can be an ethical minefield─especially regarding questions of ownership. Who owns the story, the subject or the author? The easy answer to this is the subject owns the story and the author the text, but on closer examination this may not necessarily be so. Then there are those other stakeholders who claim ownership of story: people who embrace a narrative because of its similarity to their own lives. Published stories about institutional abuse have resonated for adults who as children were placed into the care of churches or the state. Another kind of ownership is claimed by readers who accept a version of a story and dismiss any counter-story as being invalid. What I call the “Plath Phenomenon” is an example of this. I will examine these kinds of ownership of story with particular reference to the work of Janet Malcolm on biography and then will look at gaps and silences in “official” stories, those created in police stations and courtrooms.