title: Engineering and characterisation of anti-progesterone OBodies

authors: Chonira, Vikas Kariyappa

subjects: protein engineering

abstract: Molecular interactions are fundamental to communication between different parts of the cell or of an organism. These interactions can be weak and transient or strong and semi-permanent. In the case of the adaptive immune system, strong interactions between foreign antigens and specific antibodies lead to a cascade of events comprising the immune response. This phenomenon has been exploited by industry to produce high affinity binding molecules that have been used as therapeutics or diagnostics and engineered antibodies have been at the forefront of these industries. More recently, novel non-immunoglobulin binding proteins, have been similarly engineered to produce high affinity binding proteins that can potentially replace the binding function(s) of antibodies or even surpass them. The OB-fold is a high affinity binding protein domain which has previously been engineered to bind to Hen Egg-white Lysozyme (HEL) with nanomolar affinity as a proof-of-concept technology and has been given the name OBodies. This thesis explores this concept further with an OBody (D7) engineered to bind to the small molecule progesterone (P4) with potential applications to detect P4 in cow’s milk to evaluate pregnancy. Obody-P4 binding was characterised with an optimized ELISA system. This was followed by the engineering of improved versions of this OBody using phage display technology and structural characterisation of one such Obody-P4 complex using X-ray Crystallography. The three-dimensional structure provided surprising insights into the nature of the molecular interactions between P4 and the OBody. During phage display selection a new signal sequence was fortuitously discovered that provided an advantage during phage display. The new signal sequence was investigated and characterised using mutants of GFP and a DARPin sequence to uncover the nature of the selective advantage. The combination of this new signal sequence and thermostable OBody libraries further demonstrates the potential of this system to produce robust bio-sensors for diagnostic and therapeutic applications.

text: Engineering and characterisation of anti-progesterone OBodies Molecular interactions are fundamental to communication between different parts of the cell or of an organism. These interactions can be weak and transient or strong and semi-permanent. In the case of the adaptive immune system, strong interactions between foreign antigens and specific antibodies lead to a cascade of events comprising the immune response. This phenomenon has been exploited by industry to produce high affinity binding molecules that have been used as therapeutics or diagnostics and engineered antibodies have been at the forefront of these industries. More recently, novel non-immunoglobulin binding proteins, have been similarly engineered to produce high affinity binding proteins that can potentially replace the binding function(s) of antibodies or even surpass them. The OB-fold is a high affinity binding protein domain which has previously been engineered to bind to Hen Egg-white Lysozyme (HEL) with nanomolar affinity as a proof-of-concept technology and has been given the name OBodies. This thesis explores this concept further with an OBody (D7) engineered to bind to the small molecule progesterone (P4) with potential applications to detect P4 in cow’s milk to evaluate pregnancy. Obody-P4 binding was characterised with an optimized ELISA system. This was followed by the engineering of improved versions of this OBody using phage display technology and structural characterisation of one such Obody-P4 complex using X-ray Crystallography. The three-dimensional structure provided surprising insights into the nature of the molecular interactions between P4 and the OBody. During phage display selection a new signal sequence was fortuitously discovered that provided an advantage during phage display. The new signal sequence was investigated and characterised using mutants of GFP and a DARPin sequence to uncover the nature of the selective advantage. The combination of this new signal sequence and thermostable OBody libraries further demonstrates the potential of this system to produce robust bio-sensors for diagnostic and therapeutic applications.

year: 2018

title: “Mate Ni Vula”, “Masina”, “Vaʻine Fakahēʻaho”, “Eikura” Pacific sports women’s knowledge and experiences of menstruation

authors: Gibbons, Agatha

abstract: Pacific women's interest and participation in sports have increased rapidly throughout the decades. Recently, many have become competitive across a range of sports at regional, national, and international levels. Yet very little research is focused on Pacific sportswomen’s experiences, or how their cultural identities shape their performance, health, and well-being needs. This thesis focuses on Pacific sportswomen’s experiences of menstruation, and in so doing, makes an important contribution to both literature on Pacific sportswomen, and research on menstruation in sport. Research on sportswomen’s experiences of menstruation has grown significantly over recent years, but most of this research is conducted by white women on white sportswomen, with findings then generalized to all sportswomen. This thesis identifies a gap in literature, and the risks of excluding sportswomen’s cultural identities, practices, and values from research, policy, and practices focused on menstruation in sport. 
Adopting a Masi methodology, this project included three methods: a survey of 206 Pacific sportswomen; Talanoa sessions with 21 Fijian sportswomen, and interviews with 16 Pacific sportswomen living in Aotearoa New Zealand.  This project aimed to listen to, learn from, and amplify sportswomen's experiences and their understandings of menstruation, cultural beliefs, and the taboos that influence their menstrual experiences. This is a thesis with publication; some chapters are published articles, and others are manuscripts undergoing review. First, the thesis systematically identifies the gap in the literature with a scoping review of research focused on menstruation in sport, that shows that % of research ignores women’s cultural or ethnic identities. This chapter identifies the need for more culturally responsive research methods and culturally safe practices from sports and health professionals working with sportswomen from diverse backgrounds. Following this, results from the survey are presented, highlighting the diversity of cultural knowledge and practices associated with menstruation across the Pacific the survey also reveals the different cultural stigmas, beliefs, taboos, traditions, menstrual health, and sports performance of elite Pacific sportswomen. The following two chapters focus on findings from Fijian sportswomen, and then Pacific sportswomen living in Aotearoa New Zealand, with both amplifying the voices and experiences of Pacific sportswomen and their challenges of gaining culturally informed menstrual health information and support in sporting environments. These chapters describe how culture intersects with gender and other key variables (i.e., socio-economic considerations), and how these influence sportswomen’s experiences of menstruation.  
Ultimately, this research makes an important contribution to research on menstruation in sport, highlighting the need for culturally responsive methodologies and approaches in research and practice. It provides new and current cultural knowledge that can be used to improve the health, performance, and social support structures for Pacific sportswomen. This knowledge can expand sports and health providers' understanding of Pacific sportswomen's culturally specific needs, knowledge, and values concerning menstruation. Finally, this research supports the development of sport in the Pacific by providing cultural knowledge on how to engage Pacific sportswomen in safe, respectful, and constructive conversations around menstruation in sport.

text: “Mate Ni Vula”, “Masina”, “Vaʻine Fakahēʻaho”, “Eikura” Pacific sports women’s knowledge and experiences of menstruation Pacific women's interest and participation in sports have increased rapidly throughout the decades. Recently, many have become competitive across a range of sports at regional, national, and international levels. Yet very little research is focused on Pacific sportswomen’s experiences, or how their cultural identities shape their performance, health, and well-being needs. This thesis focuses on Pacific sportswomen’s experiences of menstruation, and in so doing, makes an important contribution to both literature on Pacific sportswomen, and research on menstruation in sport. Research on sportswomen’s experiences of menstruation has grown significantly over recent years, but most of this research is conducted by white women on white sportswomen, with findings then generalized to all sportswomen. This thesis identifies a gap in literature, and the risks of excluding sportswomen’s cultural identities, practices, and values from research, policy, and practices focused on menstruation in sport. 
Adopting a Masi methodology, this project included three methods: a survey of 206 Pacific sportswomen; Talanoa sessions with 21 Fijian sportswomen, and interviews with 16 Pacific sportswomen living in Aotearoa New Zealand.  This project aimed to listen to, learn from, and amplify sportswomen's experiences and their understandings of menstruation, cultural beliefs, and the taboos that influence their menstrual experiences. This is a thesis with publication; some chapters are published articles, and others are manuscripts undergoing review. First, the thesis systematically identifies the gap in the literature with a scoping review of research focused on menstruation in sport, that shows that % of research ignores women’s cultural or ethnic identities. This chapter identifies the need for more culturally responsive research methods and culturally safe practices from sports and health professionals working with sportswomen from diverse backgrounds. Following this, results from the survey are presented, highlighting the diversity of cultural knowledge and practices associated with menstruation across the Pacific the survey also reveals the different cultural stigmas, beliefs, taboos, traditions, menstrual health, and sports performance of elite Pacific sportswomen. The following two chapters focus on findings from Fijian sportswomen, and then Pacific sportswomen living in Aotearoa New Zealand, with both amplifying the voices and experiences of Pacific sportswomen and their challenges of gaining culturally informed menstrual health information and support in sporting environments. These chapters describe how culture intersects with gender and other key variables (i.e., socio-economic considerations), and how these influence sportswomen’s experiences of menstruation.  
Ultimately, this research makes an important contribution to research on menstruation in sport, highlighting the need for culturally responsive methodologies and approaches in research and practice. It provides new and current cultural knowledge that can be used to improve the health, performance, and social support structures for Pacific sportswomen. This knowledge can expand sports and health providers' understanding of Pacific sportswomen's culturally specific needs, knowledge, and values concerning menstruation. Finally, this research supports the development of sport in the Pacific by providing cultural knowledge on how to engage Pacific sportswomen in safe, respectful, and constructive conversations around menstruation in sport.

year: 2025-08

title: Indigenous perspectives and gene editing in Aotearoa New Zealand

authors: Hudson, Maui

subjects: Science & Technology

abstract: Gene editing is arguably the most significant recent addition to the modern biotechnology toolbox, bringing both profoundly challenging and enabling opportunities. From a technical point of view the specificity and relative simplicity of these new tools has broadened the potential applications. However, from an ethical point of view it has re-ignited the debates generated by earlier forms of genetic modification. In New Zealand gene editing is currently considered genetic modification and is subject to approval processes under the Environmental Protection Authority (EPA). This process requires decision makers to take into account Māori perspectives. This article outlines previously articulated Māori perspectives on genetic modification and considers the continuing influence of those cultural and ethical arguments within the new context of gene editing. It also explores the range of ways cultural values might be used to analyse the risks and benefits of gene editing in the Aotearoa New Zealand context. Methods used to obtain these perspectives consisted of (a) review of relevant literature regarding lessons learned from the responses of Maori to genetic modification, (b) interviews of selected ‘key Maori informants’ and (c) surveys of self-selected individuals from groups with interests in either genetics or environmental management. The outcomes of this pilot study identified that while Māori informants were not categorically opposed to new and emerging gene editing technologies a priori, they suggest a dynamic approach to regulation is required where specific uses or types of uses are approved on a case by case basis. This study demonstrates how the cultural cues that Māori referenced in the genetic modification debate continue to be relevant in the context of gene editing but that further work is required to characterize the strength of various positions across the broader community.

text: Indigenous perspectives and gene editing in Aotearoa New Zealand Gene editing is arguably the most significant recent addition to the modern biotechnology toolbox, bringing both profoundly challenging and enabling opportunities. From a technical point of view the specificity and relative simplicity of these new tools has broadened the potential applications. However, from an ethical point of view it has re-ignited the debates generated by earlier forms of genetic modification. In New Zealand gene editing is currently considered genetic modification and is subject to approval processes under the Environmental Protection Authority (EPA). This process requires decision makers to take into account Māori perspectives. This article outlines previously articulated Māori perspectives on genetic modification and considers the continuing influence of those cultural and ethical arguments within the new context of gene editing. It also explores the range of ways cultural values might be used to analyse the risks and benefits of gene editing in the Aotearoa New Zealand context. Methods used to obtain these perspectives consisted of (a) review of relevant literature regarding lessons learned from the responses of Maori to genetic modification, (b) interviews of selected ‘key Maori informants’ and (c) surveys of self-selected individuals from groups with interests in either genetics or environmental management. The outcomes of this pilot study identified that while Māori informants were not categorically opposed to new and emerging gene editing technologies a priori, they suggest a dynamic approach to regulation is required where specific uses or types of uses are approved on a case by case basis. This study demonstrates how the cultural cues that Māori referenced in the genetic modification debate continue to be relevant in the context of gene editing but that further work is required to characterize the strength of various positions across the broader community.

year: 2019