title: Development of a qPCR method to measure mitochondrial and genomic DNA damage with application to chemotherapy-induced DNA damage and cryopreserved cells

authors: Evans, Stephen Owen

subjects: LORD-Q

abstract: DNA damage quantitation assays such as the comet assay have focused on the measurement of total nuclear damage per cell. The adoption of PCR-based techniques to quantify DNA damage has enabled sequence- and organelle-specific assessment of DNA lesions. Here we report on an adaptation of a qPCR technique to assess DNA damage in nuclear and mitochondrial targets relative to control. Novel aspects of this assay include application of the assay to the Rotor-Gene platform with optimized DNA polymerase/fluorophore/primer set combination in a touchdown PCR protocol. Assay validation was performed using ultraviolet C radiation in A549 and THP1 cancer cell lines. A comparison was made to the comet assay applied to peripheral blood mononuclear cells, and an estimation of the effects of cryopreservation on ultraviolet C-induced DNA damage was carried out. Finally, dose responses for DNA damage were measured in peripheral blood mononuclear cells following exposure to the cytotoxic agents bleomycin and cisplatin. We show reproducible experimental outputs across the tested conditions and concordance with published findings with respect to mitochondrial and nuclear genotoxic susceptibilities. The application of this DNA damage assay to a wide range of clinical and laboratory-derived samples is both feasible and resource-efficient.

text: Development of a qPCR method to measure mitochondrial and genomic DNA damage with application to chemotherapy-induced DNA damage and cryopreserved cells DNA damage quantitation assays such as the comet assay have focused on the measurement of total nuclear damage per cell. The adoption of PCR-based techniques to quantify DNA damage has enabled sequence- and organelle-specific assessment of DNA lesions. Here we report on an adaptation of a qPCR technique to assess DNA damage in nuclear and mitochondrial targets relative to control. Novel aspects of this assay include application of the assay to the Rotor-Gene platform with optimized DNA polymerase/fluorophore/primer set combination in a touchdown PCR protocol. Assay validation was performed using ultraviolet C radiation in A549 and THP1 cancer cell lines. A comparison was made to the comet assay applied to peripheral blood mononuclear cells, and an estimation of the effects of cryopreservation on ultraviolet C-induced DNA damage was carried out. Finally, dose responses for DNA damage were measured in peripheral blood mononuclear cells following exposure to the cytotoxic agents bleomycin and cisplatin. We show reproducible experimental outputs across the tested conditions and concordance with published findings with respect to mitochondrial and nuclear genotoxic susceptibilities. The application of this DNA damage assay to a wide range of clinical and laboratory-derived samples is both feasible and resource-efficient.

year: 2016

title: Moetolo ‘sleep-crawlers’: A Samoan therapeutic approach to sexual abuse

authors: Seiuli, Byron Malaela Sotiata

abstract: Sexual abuse behaviour remains a global concern that accounts for child sexual abuse, rape and other sexual assaults. Much of these acts of violation are perpetuated, but not entirely, carried out by men against women and children. Moetolo (moetotolo) or ‘sleep crawler’ is a Samoan term that is used to describe a person who sexually violates another while they or their family are asleep. This paper provides a case study presentation and discussion with the aim of examining this challenge from a Samoan viewpoint. Insights are drawn from the authors’ psychological engagement with a sexual offender while also engaging appropriate literature to inform and provide interpretation to the case. It is through such an examination that this article seeks to make a valuable contribution to understanding patterned responses of Samoan people to sexual abuse behaviours such as moetolo, and steps to remedy such concerns within their communities.

text: Moetolo ‘sleep-crawlers’: A Samoan therapeutic approach to sexual abuse Sexual abuse behaviour remains a global concern that accounts for child sexual abuse, rape and other sexual assaults. Much of these acts of violation are perpetuated, but not entirely, carried out by men against women and children. Moetolo (moetotolo) or ‘sleep crawler’ is a Samoan term that is used to describe a person who sexually violates another while they or their family are asleep. This paper provides a case study presentation and discussion with the aim of examining this challenge from a Samoan viewpoint. Insights are drawn from the authors’ psychological engagement with a sexual offender while also engaging appropriate literature to inform and provide interpretation to the case. It is through such an examination that this article seeks to make a valuable contribution to understanding patterned responses of Samoan people to sexual abuse behaviours such as moetolo, and steps to remedy such concerns within their communities.

year: 2016

title: Mai i te kore, ki te pō, ki te ao mārama: He kohinga pūrākau hauora o ngā kaumātua Māori, Māori Elders health and wellbeing stories in life transitions

authors: Shelford, Pita

subjects: pūrākau

abstract: Introduction
New Zealand’s general population is ageing. Kaumātua (Māori elders) only make up approximately 3.1 percent of the total population and yet, they are disproportionately overrepresented in poor health, wellbeing and ageing statistics. However, this is a one-sided story and the focus of a deficit approach. There is a strong need for a strength-based approach where kaumātua voices of problem-solving and resilience related to ageing and the changes it presents, are heard. The purpose of this study is to identify and share those voices and to respond to the call from Māori academics to apply pūrākau as an alternative to Western narrative approaches. Through a pūrākau approach, this thesis sought to answer three key research questions: 

1)What are the cultural and communication features of the kaumātua teina narratives?
2) What are the cultural and communication characteristics of the kaumātua tuakana conversational styles? and 
3) What is the nature of the kaumātua journeys in how the stories unfold, and relationships develop? 

Methods
This study analyses pūrākau from the Kaumātua Mana Motuhake (KMM) collaboration project between the University of Waikato and the Rauawaawa Kaumātua Charitable Trust in Hamilton, Aotearoa New Zealand. The KMM project sought to enhance kaumātua wellbeing during life changes such as losses of licence, independence or spouse, health changes and retirement through a peer-support tuakana-teina programme. The KMM team implemented a tuakana-teina peer support programme where kaumātua (n= 121) engaged in up to three conversations as kaumātua tuakana-teina pairs. The conversations involved exploring the life changes the teina chose with the tuakana guiding the conversations and sharing potential services to help with the changes.
This thesis analysed 30 kaumātua conversations in 10 tuakana-teina pairs to tell a strengths-based story of health and wellbeing. Taking a Kaupapa Māori approach, the research prioritised, privileged and foregrounded Māori and Indigenous epistemology, ontology, and axiology. A pūrākau-grounded analysis approach was developed and applied to the conversations.

Findings
Related to the first research question, the analysis revealed kaumātua narratives that spoke to stories about adversity, turning points, independence and resilience. Regarding the second research question, the study identified key cultural and communicational characteristics kaumātua tuakana used, speaking with words of kinship and reciprocation, words of ignition and guidance, and words of empathy and affirmation. Pertaining to the third research question, the analysis illustrated conversational and relational journey types kaumātua pairs travelled, consisting of conversations that went direct to topics, detours that became defining moments, small tiki tours that came back to the topic, stories that were revisited, and hospitality becoming defining moments. 

Conclusions
The findings compiled weave a collective tapestry of kaumātua teina pūrākau, kaumātua tuakana communication styles, and kaumātua conversational and relational journeys. Kaumātua teina have voiced a pūrākau for us to understand inter-generational trauma and how cultural connection liberated their wellbeing. Kaumātua tuakana have composed a script of communication protocols emphasising connection and relationship development essential to upholding tikanga (practices, principles, communications). The paired kaumātua journeys have drawn a signposted road map that highlighted journey types, emphasising reciprocity in conversing, learning, guiding, connecting, developing, strengthening and enhancing communications, relationships, and wellbeing

Implications
Via kaumātua conversations, invaluable understandings into Māori and Indigenous wellbeing, ageing and development scholarship are offered, strengthening transformation, leadership and resilience theories. Pūrākau and whanaungatanga (relationships) methodologies are advocated for and culturally responsive research is promoted. Health, wellbeing and ageing programme development, policy and service delivery can be practically informed, accentuating mentorship, intergenerational initiatives, and culturally appropriate kaumātua support. Together, the findings can cultivate community wellbeing and nurture cultural continuity.

text: Mai i te kore, ki te pō, ki te ao mārama: He kohinga pūrākau hauora o ngā kaumātua Māori, Māori Elders health and wellbeing stories in life transitions Introduction
New Zealand’s general population is ageing. Kaumātua (Māori elders) only make up approximately 3.1 percent of the total population and yet, they are disproportionately overrepresented in poor health, wellbeing and ageing statistics. However, this is a one-sided story and the focus of a deficit approach. There is a strong need for a strength-based approach where kaumātua voices of problem-solving and resilience related to ageing and the changes it presents, are heard. The purpose of this study is to identify and share those voices and to respond to the call from Māori academics to apply pūrākau as an alternative to Western narrative approaches. Through a pūrākau approach, this thesis sought to answer three key research questions: 

1)What are the cultural and communication features of the kaumātua teina narratives?
2) What are the cultural and communication characteristics of the kaumātua tuakana conversational styles? and 
3) What is the nature of the kaumātua journeys in how the stories unfold, and relationships develop? 

Methods
This study analyses pūrākau from the Kaumātua Mana Motuhake (KMM) collaboration project between the University of Waikato and the Rauawaawa Kaumātua Charitable Trust in Hamilton, Aotearoa New Zealand. The KMM project sought to enhance kaumātua wellbeing during life changes such as losses of licence, independence or spouse, health changes and retirement through a peer-support tuakana-teina programme. The KMM team implemented a tuakana-teina peer support programme where kaumātua (n= 121) engaged in up to three conversations as kaumātua tuakana-teina pairs. The conversations involved exploring the life changes the teina chose with the tuakana guiding the conversations and sharing potential services to help with the changes.
This thesis analysed 30 kaumātua conversations in 10 tuakana-teina pairs to tell a strengths-based story of health and wellbeing. Taking a Kaupapa Māori approach, the research prioritised, privileged and foregrounded Māori and Indigenous epistemology, ontology, and axiology. A pūrākau-grounded analysis approach was developed and applied to the conversations.

Findings
Related to the first research question, the analysis revealed kaumātua narratives that spoke to stories about adversity, turning points, independence and resilience. Regarding the second research question, the study identified key cultural and communicational characteristics kaumātua tuakana used, speaking with words of kinship and reciprocation, words of ignition and guidance, and words of empathy and affirmation. Pertaining to the third research question, the analysis illustrated conversational and relational journey types kaumātua pairs travelled, consisting of conversations that went direct to topics, detours that became defining moments, small tiki tours that came back to the topic, stories that were revisited, and hospitality becoming defining moments. 

Conclusions
The findings compiled weave a collective tapestry of kaumātua teina pūrākau, kaumātua tuakana communication styles, and kaumātua conversational and relational journeys. Kaumātua teina have voiced a pūrākau for us to understand inter-generational trauma and how cultural connection liberated their wellbeing. Kaumātua tuakana have composed a script of communication protocols emphasising connection and relationship development essential to upholding tikanga (practices, principles, communications). The paired kaumātua journeys have drawn a signposted road map that highlighted journey types, emphasising reciprocity in conversing, learning, guiding, connecting, developing, strengthening and enhancing communications, relationships, and wellbeing

Implications
Via kaumātua conversations, invaluable understandings into Māori and Indigenous wellbeing, ageing and development scholarship are offered, strengthening transformation, leadership and resilience theories. Pūrākau and whanaungatanga (relationships) methodologies are advocated for and culturally responsive research is promoted. Health, wellbeing and ageing programme development, policy and service delivery can be practically informed, accentuating mentorship, intergenerational initiatives, and culturally appropriate kaumātua support. Together, the findings can cultivate community wellbeing and nurture cultural continuity.

year: 2025-11-04