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: Shareholders’ requirements for corporate environmental disclosures: A cross country comparison

authors: de Villiers, Charl

subjects: Shareholders’ opinions

abstract: e survey individual shareholders in Australia, the UK and the US regarding corporate environmental disclosures. In general, respondents in the three countries are interested in, and positively disposed towards, these disclosures. We observe country and gender differences with Australian and female respondents more in favour of environmental reporting than others. Specifically, respondents require disclosure of an overview of environmental risks and impacts, the environmental policy, performance against measurable environmental targets and information on a range of environmental costs. Most shareholders require environmental disclosures to be audited. Shareholders call for environmental information because they believe managers should be accountable to shareholders for their companies’ environmental impacts. Furthermore, shareholders have indicated the uses for specific types of environmental information. Our results imply that legislators, standard setters and companies have to consider the policy implications of these shareholder views.

text: Shareholders’ requirements for corporate environmental disclosures: A cross country comparison e survey individual shareholders in Australia, the UK and the US regarding corporate environmental disclosures. In general, respondents in the three countries are interested in, and positively disposed towards, these disclosures. We observe country and gender differences with Australian and female respondents more in favour of environmental reporting than others. Specifically, respondents require disclosure of an overview of environmental risks and impacts, the environmental policy, performance against measurable environmental targets and information on a range of environmental costs. Most shareholders require environmental disclosures to be audited. Shareholders call for environmental information because they believe managers should be accountable to shareholders for their companies’ environmental impacts. Furthermore, shareholders have indicated the uses for specific types of environmental information. Our results imply that legislators, standard setters and companies have to consider the policy implications of these shareholder views.

year: 2010

title: Ascending the Poutama: Culturally responsive diabetes care for Māori communities

authors: Crosswell, Rebekah Jane

subjects: culturally responsive

abstract: Introduction
Type 2 diabetes (T2D) significantly impacts Māori populations in Aotearoa New Zealand (NZ) who experience health disparities including suboptimal disease management.

Aim
To explore culturally responsive approaches to supporting Māori living with T2D from the perspective of healthcare providers.

Methods
Nine semi-structured interviews were conducted with clinicians from various disciplines (general practitioner, podiatrist, exercise physiologist, three kaiāwhina, dietitian, two pharmacists); five clinicians identify as Māori, four as non-Māori. Kaupapa Māori inductive thematic analysis was used.

Results
Four key themes emerged: empowering patient-centred communication, whānau involvement, interprofessional collaboration, and culturally appropriate analogies.

Discussion
The findings underscore the need for holistic, culturally responsive T2D care that prioritises culturally appropriate communication to improve patient engagement, health outcomes and address Māori health disparities.

text: Ascending the Poutama: Culturally responsive diabetes care for Māori communities Introduction
Type 2 diabetes (T2D) significantly impacts Māori populations in Aotearoa New Zealand (NZ) who experience health disparities including suboptimal disease management.

Aim
To explore culturally responsive approaches to supporting Māori living with T2D from the perspective of healthcare providers.

Methods
Nine semi-structured interviews were conducted with clinicians from various disciplines (general practitioner, podiatrist, exercise physiologist, three kaiāwhina, dietitian, two pharmacists); five clinicians identify as Māori, four as non-Māori. Kaupapa Māori inductive thematic analysis was used.

Results
Four key themes emerged: empowering patient-centred communication, whānau involvement, interprofessional collaboration, and culturally appropriate analogies.

Discussion
The findings underscore the need for holistic, culturally responsive T2D care that prioritises culturally appropriate communication to improve patient engagement, health outcomes and address Māori health disparities.

year: 2025