title: Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry

authors: McKenzie, Joanna Leigh

subjects: Mycobacteria

abstract: The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press.

text: Determination of ribonuclease sequence-specificity using Pentaprobes and mass spectrometry The VapBC toxin-antitoxin (TA) family is the largest of nine identified TA families. The toxin, VapC, is a metal-dependent ribonuclease that is inhibited by its cognate antitoxin, VapB. Although the VapBCs are the largest TA family, little is known about their biological roles. Here we describe a new general method for the overexpression and purification of toxic VapC proteins and subsequent determination of their RNase sequence-specificity. Functional VapC was isolated by expression of the nontoxic VapBC complex, followed by removal of the labile antitoxin (VapB) using limited trypsin digestion. We have then developed a sensitive and robust method for determining VapC ribonuclease sequence-specificity. This technique employs the use of Pentaprobes as substrates for VapC. These are RNA sequences encoding every combination of five bases. We combine the RNase reaction with MALDI-TOF MS to detect and analyze the cleavage products and thus determine the RNA cut sites. Successful MALDI-TOF MS analysis of RNA fragments is acutely dependent on sample preparation methods. The sequencespecificity of four VapC proteins from two different organisms (VapCPAE0151 and VapCPAE2754 from Pyrobaculum aerophilum, and VapCRv0065 and VapCRv0617 from Mycobacterium tuberculosis) was successfully determined using the described strategy. This rapid and sensitive method can be applied to determine the sequence-specificity of VapC ribonucleases along with other RNA interferases (such as MazF) from a range of organisms. Published by Cold Spring Harbor Laboratory Press.

year: 2012

title: The role of Adaptive Energy Digital Twin technology in decarbonising emissions in the New Zealand process heat sector

authors: Walmsley, Michael R.W.

subjects: process heat decarbonisation

abstract: New Zealand (NZ) is uniquely placed to meet the goal of net zero emissions in the industry sector by 2050, but only with an 80% expansion of renewable electricity generation (TWh) combined with energy efficiency improvements through advanced process heat integration, improved energy system design and control, future deployment of high temperature heat pumps (HTHP) for process heat up to 200oC and biomass for heat above 200oC, and smart integration of renewable electricity via microgrid connected factories and communities. The development of Adaptive Energy Digital Twin (AEDT) technology applied widely by energy consultants, equipment providers and factory owners, is considered a key development to aid industry decarbonisation. A seven-year research programme is underway in NZ to develop an open source AEDT technology platform and to demonstrate its usefulness for decarbonising a wide variety of industry sectors, such as dairy, meat, food, wood, paper, metals and chemical processing industries of NZ. Electricity supply and demand differences across the regions of NZ for 2020 and predicted for 2050 are also presented.

text: The role of Adaptive Energy Digital Twin technology in decarbonising emissions in the New Zealand process heat sector New Zealand (NZ) is uniquely placed to meet the goal of net zero emissions in the industry sector by 2050, but only with an 80% expansion of renewable electricity generation (TWh) combined with energy efficiency improvements through advanced process heat integration, improved energy system design and control, future deployment of high temperature heat pumps (HTHP) for process heat up to 200oC and biomass for heat above 200oC, and smart integration of renewable electricity via microgrid connected factories and communities. The development of Adaptive Energy Digital Twin (AEDT) technology applied widely by energy consultants, equipment providers and factory owners, is considered a key development to aid industry decarbonisation. A seven-year research programme is underway in NZ to develop an open source AEDT technology platform and to demonstrate its usefulness for decarbonising a wide variety of industry sectors, such as dairy, meat, food, wood, paper, metals and chemical processing industries of NZ. Electricity supply and demand differences across the regions of NZ for 2020 and predicted for 2050 are also presented.

year: 2023

title: The Māori precariat: A silhouette

authors: Stubbs, Thomas

abstract: Recent financial crises and a host of punitive labour and welfare reforms have intensified socio-economic divisions across advanced nations. In Aotearoa New Zealand, the indigenous Maori peoples are subject to systemic labour market disadvantages, and are thus more likely to be affected by increased inequalities in times of economic downturn. Despite the legacies of systematic economic exclusion and discrimination in the economy, the majority of Maori demonstrate a resounding level of social resilience in the face of this hostile context. Even so, there is evidence to suggest that an unacceptable number of Maori still struggle to overcome these challenges. For instance, 13.0 per cent of Maori are in temporary work compared to 8.2 per cent of Pakeha,1 and 12.4 per cent of Maori are experiencing unemployment against 4.4 per cent. of Pakeha.2 These trends contribute to a range of socio-economic challenges, to which mainstream and Maori service providers are struggling to respond.
In this chapter, we develop a demographic silhouette to seek an understanding of socio-economically marginalised - or 'precariat' - Maori. Although we are wary of portraying Maori in an unduly negative light, we also submit that an appreciation of the extent of the issues faced by this group is necessary if our analysis is to be useful in the development of an effective response. We begin by defining the precariat. Using data obtained from Statistics New Zealand's Te Kupenga survey of Maori wellbeing, we then outline the prevalence and composition of the Maori precariat in Aotearoa New Zealand. Overall, we find that the Maori precariat comprises a substantial portion of the Maori population, particularly among females, younger age groups, and those with low or no qualifications. They are predominantly located-in regions that have high levels of seasonal employment, and in more deprived areas. We also find the value attached to Maori culture and experiences of stigma for this group are, for the most part, similar to those among the Maori non-precariat.

text: The Māori precariat: A silhouette Recent financial crises and a host of punitive labour and welfare reforms have intensified socio-economic divisions across advanced nations. In Aotearoa New Zealand, the indigenous Maori peoples are subject to systemic labour market disadvantages, and are thus more likely to be affected by increased inequalities in times of economic downturn. Despite the legacies of systematic economic exclusion and discrimination in the economy, the majority of Maori demonstrate a resounding level of social resilience in the face of this hostile context. Even so, there is evidence to suggest that an unacceptable number of Maori still struggle to overcome these challenges. For instance, 13.0 per cent of Maori are in temporary work compared to 8.2 per cent of Pakeha,1 and 12.4 per cent of Maori are experiencing unemployment against 4.4 per cent. of Pakeha.2 These trends contribute to a range of socio-economic challenges, to which mainstream and Maori service providers are struggling to respond.
In this chapter, we develop a demographic silhouette to seek an understanding of socio-economically marginalised - or 'precariat' - Maori. Although we are wary of portraying Maori in an unduly negative light, we also submit that an appreciation of the extent of the issues faced by this group is necessary if our analysis is to be useful in the development of an effective response. We begin by defining the precariat. Using data obtained from Statistics New Zealand's Te Kupenga survey of Maori wellbeing, we then outline the prevalence and composition of the Maori precariat in Aotearoa New Zealand. Overall, we find that the Maori precariat comprises a substantial portion of the Maori population, particularly among females, younger age groups, and those with low or no qualifications. They are predominantly located-in regions that have high levels of seasonal employment, and in more deprived areas. We also find the value attached to Maori culture and experiences of stigma for this group are, for the most part, similar to those among the Maori non-precariat.

year: 2017