title: Optimisation of bovine embryonic stem cell-like line derivation and candidate gene expression analysis

authors: Reddy, Shobana

abstract: The successful derivation of bovine embryonic stem cells (bESCs) has been an area of interest for livestock scientists over the years. These cells possess significant potential for revolutionising livestock breeding by enabling accelerated genetic gain within a single generation. Although robust derivation methods exist for mouse and human ESCs (mESC, hESC), this is not the case for livestock species. Its derivation remains challenging due to the lack of understanding of signalling pathways that maintain pluripotency and regulate early embryo development in livestock species, along with the failure of direct application of mESC and hESC culture conditions. In recent years, studies have reported the development of culture conditions that support bESC derivation, which were adapted in this thesis. This study aims to optimise bESC culture conditions, as well as optimise quantitative PCR methods for gene expression analysis to assess pluripotency. Using cell enrichment treatments, differential plating, colony picking and cell straining, three bESC-like cell lines were derived, which included a wild-type cell line (17), a red fluorescent reporter cell line (R23), and a transgenic β-κ-casein cell line (B2). Although these cell lines were found to express pluripotency markers, both gene and protein expression analysis revealed that the cell lines comprised of a mixed population. Further validation is required to confirm the identity of the derived cells, including using single cell RNA sequencing, karyotyping, in vitro and in vivo differentiation, whole animal generation, and germline contribution assessment.

text: Optimisation of bovine embryonic stem cell-like line derivation and candidate gene expression analysis The successful derivation of bovine embryonic stem cells (bESCs) has been an area of interest for livestock scientists over the years. These cells possess significant potential for revolutionising livestock breeding by enabling accelerated genetic gain within a single generation. Although robust derivation methods exist for mouse and human ESCs (mESC, hESC), this is not the case for livestock species. Its derivation remains challenging due to the lack of understanding of signalling pathways that maintain pluripotency and regulate early embryo development in livestock species, along with the failure of direct application of mESC and hESC culture conditions. In recent years, studies have reported the development of culture conditions that support bESC derivation, which were adapted in this thesis. This study aims to optimise bESC culture conditions, as well as optimise quantitative PCR methods for gene expression analysis to assess pluripotency. Using cell enrichment treatments, differential plating, colony picking and cell straining, three bESC-like cell lines were derived, which included a wild-type cell line (17), a red fluorescent reporter cell line (R23), and a transgenic β-κ-casein cell line (B2). Although these cell lines were found to express pluripotency markers, both gene and protein expression analysis revealed that the cell lines comprised of a mixed population. Further validation is required to confirm the identity of the derived cells, including using single cell RNA sequencing, karyotyping, in vitro and in vivo differentiation, whole animal generation, and germline contribution assessment.

year: 2025

title: A framework for compositional nonblocking verification of extended finite-state machines

authors: Mohajerani, Sahar

subjects: Science & Technology

abstract: This paper presents a framework for compositional nonblocking verification of discrete event systems modelled as extended finite-state machines (EFSM). Previous results are improved to consider general conflict-equivalence based abstractions of EFSMs communicating both via shared variables and events. Performance issues resulting from the conversion of EFSM systems to finite-state machine systems are avoided by operating directly on EFSMs, deferring the unfolding of variables into state machines as long as possible. Several additional methods to abstract EFSMs and remove events are also presented. The proposed algorithm has been implemented in the discrete event systems tool Supremica, and the paper presents experimental results for several large EFSM models that can be verified faster than by previously used methods.

text: A framework for compositional nonblocking verification of extended finite-state machines This paper presents a framework for compositional nonblocking verification of discrete event systems modelled as extended finite-state machines (EFSM). Previous results are improved to consider general conflict-equivalence based abstractions of EFSMs communicating both via shared variables and events. Performance issues resulting from the conversion of EFSM systems to finite-state machine systems are avoided by operating directly on EFSMs, deferring the unfolding of variables into state machines as long as possible. Several additional methods to abstract EFSMs and remove events are also presented. The proposed algorithm has been implemented in the discrete event systems tool Supremica, and the paper presents experimental results for several large EFSM models that can be verified faster than by previously used methods.

year: 2016-03-01

title: Joe Gillayley: A model of cultural hybridity in Keri Hulme’s novel The Bone People

authors: Cozma, Codrina

subjects: Literary analysis

abstract: Born and raised at the confluence of two cultures, Western and Māori, the New Zealand author Keri Hulme crafts in her 1983 novel, The Bone People, a vigorous literary discourse whose integrity and realism often topple consecrated stereotyping colonial theories. Joe Gillayley displays complex mechanisms of cultural hybridity in major areas of his private and social life. His initially ambivalent attitude towards both his Western and his Māori heritage are evident in his personal, social and economic life - in his interactions with his family, his semi-adopted son Simon/ Himi, his friend Kerewin, his work, school authorities, and tribal figures such as the mysterious prophet Tiaki Mira. Post-colonial scholarship provides the context in which the analysis provided here is conducted, an analysis that highlights the futility of colonial theories that are based on distorted, stereotypical images of both Western and indigenous life and values.

text: Joe Gillayley: A model of cultural hybridity in Keri Hulme’s novel The Bone People Born and raised at the confluence of two cultures, Western and Māori, the New Zealand author Keri Hulme crafts in her 1983 novel, The Bone People, a vigorous literary discourse whose integrity and realism often topple consecrated stereotyping colonial theories. Joe Gillayley displays complex mechanisms of cultural hybridity in major areas of his private and social life. His initially ambivalent attitude towards both his Western and his Māori heritage are evident in his personal, social and economic life - in his interactions with his family, his semi-adopted son Simon/ Himi, his friend Kerewin, his work, school authorities, and tribal figures such as the mysterious prophet Tiaki Mira. Post-colonial scholarship provides the context in which the analysis provided here is conducted, an analysis that highlights the futility of colonial theories that are based on distorted, stereotypical images of both Western and indigenous life and values.

year: 2004-02