title: A modular plasmid system for expression of genes in Neisseria gonorrhoeae

authors: Flemming, Chloe Dale

subjects: Plasmid

abstract: Neisseria gonorrhoeae is the causative agent of the sexually transmitted infection, gonorrhoea. Antimicrobial resistance in N. gonorrhoeae is of increasing concern and there is no current vaccine. Over the last 80 years, N. gonorrhoeae has become highly resistant to many frontline antibiotics, and as such, there are now limited treatment options. Research and development is critical for the discovery of new diagnostic assays, antimicrobials, and vaccines that combat N. gonorrhoeae. 
To date, there are a variety of genetic tools available for generating mutations, gene deletions, and gene complementation in N. gonorrhoeae. Plasmids are available for the introduction of new genes of interest and complementation, however replicating plasmids are often unstable, time consuming to manipulate, and suffer from poor transformation efficiencies.

Here, this project aimed to resolve the current issues with N. gonorrhoeae plasmid systems by using a Golden Gate assembly technique to design a modular cloning system specifically to enable custom gene expression in N. gonorrhoeae. Golden Gate cloning uses Type IIS restriction endonucleases to allow the assembly of complex multipart plasmids from reusable DNA parts. These include the gonococcal-specific cryptic plasmid maintenance region from pEG2, the Opa promoter, antibiotic resistance genes encoding kanamycin and erythromycin resistance, fluorescent proteins, and a ColE1 origin of replication for propagation in Escherichia coli. Our system was designed to be more efficient by enabling the rapid generation of very small, customised plasmids capable of replication in both E. coli and N. gonorrhoeae.
Currently, there are vast gaps in our knowledge of N. gonorrhoeae biology. This thesis offers a new system that is an efficient tool for custom gene expression in N. gonorrhoeae to improve our understanding of this pathogen. Improving research approaches, such as mutagenesis and complementation, will contribute to drug and vaccine discovery efforts searching for reliable treatments for this devastating pathogen.

text: A modular plasmid system for expression of genes in Neisseria gonorrhoeae Neisseria gonorrhoeae is the causative agent of the sexually transmitted infection, gonorrhoea. Antimicrobial resistance in N. gonorrhoeae is of increasing concern and there is no current vaccine. Over the last 80 years, N. gonorrhoeae has become highly resistant to many frontline antibiotics, and as such, there are now limited treatment options. Research and development is critical for the discovery of new diagnostic assays, antimicrobials, and vaccines that combat N. gonorrhoeae. 
To date, there are a variety of genetic tools available for generating mutations, gene deletions, and gene complementation in N. gonorrhoeae. Plasmids are available for the introduction of new genes of interest and complementation, however replicating plasmids are often unstable, time consuming to manipulate, and suffer from poor transformation efficiencies.

Here, this project aimed to resolve the current issues with N. gonorrhoeae plasmid systems by using a Golden Gate assembly technique to design a modular cloning system specifically to enable custom gene expression in N. gonorrhoeae. Golden Gate cloning uses Type IIS restriction endonucleases to allow the assembly of complex multipart plasmids from reusable DNA parts. These include the gonococcal-specific cryptic plasmid maintenance region from pEG2, the Opa promoter, antibiotic resistance genes encoding kanamycin and erythromycin resistance, fluorescent proteins, and a ColE1 origin of replication for propagation in Escherichia coli. Our system was designed to be more efficient by enabling the rapid generation of very small, customised plasmids capable of replication in both E. coli and N. gonorrhoeae.
Currently, there are vast gaps in our knowledge of N. gonorrhoeae biology. This thesis offers a new system that is an efficient tool for custom gene expression in N. gonorrhoeae to improve our understanding of this pathogen. Improving research approaches, such as mutagenesis and complementation, will contribute to drug and vaccine discovery efforts searching for reliable treatments for this devastating pathogen.

year: 2023

title: 'Mong'st the furies finde just recompence': Suicide and the supernatural in William Sampson's The Vow Breaker (1636)

authors: Martin, Fiona

abstract: The prevailing view on suicide in early modern England was that it was absolutely 'contrary to the Lawes and ordinances of God: 1 and contemporary theologians including John Sym, William Willymat, Robert Hill and Richard Greenham expounded in sermons and treatises on the prohibitions against self-murder and the dire consequences attendant upon the soul of the deceased.2 To take one's own life indirectly challenged the authority of the Church, and suicide, in its violation of community values, also awakened fears of the supernatural and of what the condemned soul might be capable. On the popular stage, of course, dramatizations of suicide could provide shocking but arresting scenes of conflict, both on the personal level and within the affected community.

text: 'Mong'st the furies finde just recompence': Suicide and the supernatural in William Sampson's The Vow Breaker (1636) The prevailing view on suicide in early modern England was that it was absolutely 'contrary to the Lawes and ordinances of God: 1 and contemporary theologians including John Sym, William Willymat, Robert Hill and Richard Greenham expounded in sermons and treatises on the prohibitions against self-murder and the dire consequences attendant upon the soul of the deceased.2 To take one's own life indirectly challenged the authority of the Church, and suicide, in its violation of community values, also awakened fears of the supernatural and of what the condemned soul might be capable. On the popular stage, of course, dramatizations of suicide could provide shocking but arresting scenes of conflict, both on the personal level and within the affected community.

year: 2015

title: An Evaluation of an Impaired Driver Treatment Programme Facilitated by Tūhoe Hauora

authors: Waru, Nicole Arihia

subjects: Tuhoe Hauora

abstract: Driving while impaired is a substantial issue in Aotearoa.  It has the potential to result in loss of life and cause significant health and economic strain on society.  Recidivist impaired drivers are major contributors to this problem.  Countermeasures of impaired driving have previously included liquor licencing, drinking age restrictions, health promotion campaigns and health rehabilitation therapies.  The Ministry of Health offer an Impaired Driving therapy throughout Aotearoa as countermeasure to this issue.  The programme is administered through the funder-contract system. Participation is compulsory for those mandated by court order.  
This study evaluates an Impaired Driver programme facilitated by Tūhoe Hauora; a kaupapa Māori mental health and addiction provider based in Taneatua, Eastern Bay of Plenty.  The course is delivered once weekly over a period of ten weeks to a group of twelve or less recidivist impaired drivers or first time offenders who were convicted of driving with excessive levels of breath alcohol.    
The evaluation draws from a community psychology paradigm in exploration of the lived experiences of programme participants, in the context of course 2 of the 2014 Tūhoe Hauora Impaired Driving course.  
The experiences of five individuals were explored using in-depth face to face interviews and presented using a case-by-case approach.  Participant observation, document review, evaluation visits and participant surveys were used as supplement methods of inquiry.  The findings illustrated that Tūhoe Hauora met contract requirements by addressing impaired driving recidivism to date of interview.  The evaluation also found the course actualising the mission of Tūhoe Hauora which was to address the holistic well-being of programme participants.  The research did present areas in the programme that could be improved particularly pertaining to the referral process and a need for agency collaboration.  Recommendations were formulated in attempt to address the programme gaps and discussion is offered in addressing the multi-layered oppression that was experienced throughout the evaluation process by both Tūhoe Hauora and the evaluation participants.

text: An Evaluation of an Impaired Driver Treatment Programme Facilitated by Tūhoe Hauora Driving while impaired is a substantial issue in Aotearoa.  It has the potential to result in loss of life and cause significant health and economic strain on society.  Recidivist impaired drivers are major contributors to this problem.  Countermeasures of impaired driving have previously included liquor licencing, drinking age restrictions, health promotion campaigns and health rehabilitation therapies.  The Ministry of Health offer an Impaired Driving therapy throughout Aotearoa as countermeasure to this issue.  The programme is administered through the funder-contract system. Participation is compulsory for those mandated by court order.  
This study evaluates an Impaired Driver programme facilitated by Tūhoe Hauora; a kaupapa Māori mental health and addiction provider based in Taneatua, Eastern Bay of Plenty.  The course is delivered once weekly over a period of ten weeks to a group of twelve or less recidivist impaired drivers or first time offenders who were convicted of driving with excessive levels of breath alcohol.    
The evaluation draws from a community psychology paradigm in exploration of the lived experiences of programme participants, in the context of course 2 of the 2014 Tūhoe Hauora Impaired Driving course.  
The experiences of five individuals were explored using in-depth face to face interviews and presented using a case-by-case approach.  Participant observation, document review, evaluation visits and participant surveys were used as supplement methods of inquiry.  The findings illustrated that Tūhoe Hauora met contract requirements by addressing impaired driving recidivism to date of interview.  The evaluation also found the course actualising the mission of Tūhoe Hauora which was to address the holistic well-being of programme participants.  The research did present areas in the programme that could be improved particularly pertaining to the referral process and a need for agency collaboration.  Recommendations were formulated in attempt to address the programme gaps and discussion is offered in addressing the multi-layered oppression that was experienced throughout the evaluation process by both Tūhoe Hauora and the evaluation participants.

year: 2016