title: Caldalkalibacillus thermarum PhoH2: Solving the solubility puzzle

authors: Dillon, Brooke Jane

subjects: Caldalkalibacillus thermarum

abstract: Previous attempts to determine the structure of the protein PhoH2 from Mycobacterium tuberculosis, Mycobacterium smegmatis and Thermobispora bispora have been unsuccessful producing diffraction data to low resolution. The thermophilic organism Caldalkibacillus thermarum was thought to be an advantageous option from which to clone, express and purify PhoH2 from for the purpose of structure determination. PhoH2 consists of two domains, an N terminal PIN domain, known for its toxic properties as part of many toxin-antitoxin systems present in M. tuberculosis, and a C-terminal PhoH domain, an RNA helicase suspected to be involved in phosphate starvation responses. The cloning and expression of three variations of recombinant C. thermarum PhoH2 was successful in Escherichia coli. However the purification of PhoH2 continues to yield insoluble protein, despite the range of purification buffer conditions screened, thus preventing downstream biochemical characterisation and structural investigations. Multiple options exist to overcome this insolubility problem; including alternate plasmids for protein expression and purification that alter the tag location and type such as C-terminal His-tags and alternative fusion tags.

text: Caldalkalibacillus thermarum PhoH2: Solving the solubility puzzle Previous attempts to determine the structure of the protein PhoH2 from Mycobacterium tuberculosis, Mycobacterium smegmatis and Thermobispora bispora have been unsuccessful producing diffraction data to low resolution. The thermophilic organism Caldalkibacillus thermarum was thought to be an advantageous option from which to clone, express and purify PhoH2 from for the purpose of structure determination. PhoH2 consists of two domains, an N terminal PIN domain, known for its toxic properties as part of many toxin-antitoxin systems present in M. tuberculosis, and a C-terminal PhoH domain, an RNA helicase suspected to be involved in phosphate starvation responses. The cloning and expression of three variations of recombinant C. thermarum PhoH2 was successful in Escherichia coli. However the purification of PhoH2 continues to yield insoluble protein, despite the range of purification buffer conditions screened, thus preventing downstream biochemical characterisation and structural investigations. Multiple options exist to overcome this insolubility problem; including alternate plasmids for protein expression and purification that alter the tag location and type such as C-terminal His-tags and alternative fusion tags.

year: 2017

title: The Thames high school endowment at Waiorongomai

authors: Hart, Philip

abstract: Before it was known that gold existed nearby, the government selected a block of land at Waiorongomai as an endowment to fund a proposed high school at Thames. Sections of varying sizes and quality were surveyed and leased, at what were commonly viewed as excessive rates, and for decades lessees requested reductions and on many occasions asked the government to take over the endowment. As the goldfield faded, some small farmers abandoned their sections but others succeeded despite being under-capitalized. In the twentieth century, noxious weeds created on-going problems for those remaining on the land.
The high school board, which supervised the farmers, was accused of not assisting them, notably by not providing access roads. The endowment was never as profitable for the high school as had been anticipated, and in 1949 it, like all similar endowments, was abolished.

text: The Thames high school endowment at Waiorongomai Before it was known that gold existed nearby, the government selected a block of land at Waiorongomai as an endowment to fund a proposed high school at Thames. Sections of varying sizes and quality were surveyed and leased, at what were commonly viewed as excessive rates, and for decades lessees requested reductions and on many occasions asked the government to take over the endowment. As the goldfield faded, some small farmers abandoned their sections but others succeeded despite being under-capitalized. In the twentieth century, noxious weeds created on-going problems for those remaining on the land.
The high school board, which supervised the farmers, was accused of not assisting them, notably by not providing access roads. The endowment was never as profitable for the high school as had been anticipated, and in 1949 it, like all similar endowments, was abolished.

year: 2016

title: Countering gentrification in Tāmaki Makarau: Towards a framework for urban environmental justice

authors: Terruhn, Jessica

abstract: This policy brief argues for urban policies that actively aim to end gentrification.

Gentrification is intimately tied to state-led processes of urban renewal. Though historically understood as the displacement of low-income households as an unfortunate byproduct of socioeconomic and demographic neighbourhood change, contemporary gentrification is constituted by profit-driven investment decisions that erase undervalued neighbourhoods and their communities in the name of urban renewal (variously referred to as redevelopment, regeneration, renaissance, or revitalisation).

These processes of gentrification fundamentally contribute to environmental injustice. At the same time as creating ‘zones of abundance’, neighbourhood regeneration projects simultaneously (re-)create ‘zones of neglect and sacrifice’, which are burdened with environmental risks. As low-income and ethnic minority households are more likely to either reside in or to be displaced into these neighbourhoods, they are disproportionately exposed to environmental risk factors.

Moreover, gentrification enables and legitimises the exclusion of marginalised communities in multiple ways: 1) it devalues existing places and their residents in favour of regeneration, 2) it displaces longtime residents from regenerated neighbourhoods because of increasing property values, and/or 3) it alienates longtime residents as neighbourhood aesthetics, amenities and services change and become unaffordable and/or irrelevant.

As gentrification contributes to low-income households’ experiences of precarity and severs community ties and place attachments, it generates significant costs to personal and community health and wellbeing as well as social cohesion. The recommendations articulated in this brief emphasise the importance of transformative policy solutions that actively counter gentrification.
These include:
• Equitable investment in urban infrastructure
• Prioritising affordable housing
• Ensuring accessible transportation
• Empowering local communities
• Implementing a Te Tiriti o Waitangi approach to policy development

text: Countering gentrification in Tāmaki Makarau: Towards a framework for urban environmental justice This policy brief argues for urban policies that actively aim to end gentrification.

Gentrification is intimately tied to state-led processes of urban renewal. Though historically understood as the displacement of low-income households as an unfortunate byproduct of socioeconomic and demographic neighbourhood change, contemporary gentrification is constituted by profit-driven investment decisions that erase undervalued neighbourhoods and their communities in the name of urban renewal (variously referred to as redevelopment, regeneration, renaissance, or revitalisation).

These processes of gentrification fundamentally contribute to environmental injustice. At the same time as creating ‘zones of abundance’, neighbourhood regeneration projects simultaneously (re-)create ‘zones of neglect and sacrifice’, which are burdened with environmental risks. As low-income and ethnic minority households are more likely to either reside in or to be displaced into these neighbourhoods, they are disproportionately exposed to environmental risk factors.

Moreover, gentrification enables and legitimises the exclusion of marginalised communities in multiple ways: 1) it devalues existing places and their residents in favour of regeneration, 2) it displaces longtime residents from regenerated neighbourhoods because of increasing property values, and/or 3) it alienates longtime residents as neighbourhood aesthetics, amenities and services change and become unaffordable and/or irrelevant.

As gentrification contributes to low-income households’ experiences of precarity and severs community ties and place attachments, it generates significant costs to personal and community health and wellbeing as well as social cohesion. The recommendations articulated in this brief emphasise the importance of transformative policy solutions that actively counter gentrification.
These include:
• Equitable investment in urban infrastructure
• Prioritising affordable housing
• Ensuring accessible transportation
• Empowering local communities
• Implementing a Te Tiriti o Waitangi approach to policy development

year: 2025