title: A promoter characterisation of the Ets transcription factor gene, Ese-2/Elf-5

authors: Speed, Kathy Ann

abstract: E26 transformation specific (Ets) proteins are a versatile class of transcription factors. They act both up and downstream of signal transduction cascades, are causative in cancer generation and are able to control the expression of other transcription factors. Thus they play an important role in cellular growth, proliferation and differentiation. The ets gene family is characterised by an 85 amino acid DNA binding domain and currently consists of around 30 members. They have been shown to be proto-oncogenes and it is this feature that has elicited much research into their regulation and function in both normal and neoplasic tissue. The high level of conservation within the ETS domain of the family identifies all Ets proteins as sequence-specific DNA-binding proteins. Many have been identified as having a role in the regulation of gene expression. The retention of many ets genes throughout evolution implies there would be few redundant genes (Thomas, 1993) and therefore each gene would have a unique biological role. Because of the common DNA binding domain, the regulation of the ets genes must be complex to provide specificity of action throughout the family (Laudet et al., 1999).

A cDNA sequence was isolated from mammary gland tissue and characterised as belonging to the Ets family of transcription factors. The gene was designated BrETS for Breast ETS. Recent publications have described the gene as Elf-5 (Zhou et al., 1998) and ESE-2 (Oettgen et al., 1999). BrEts showed a restricted pattern of spatial expression, being strongly expressed in the mammary gland. Its expression in the mammary gland also exhibits a tightly controlled temporal pattern, which implicates it in mammary processes such as development and differentiation. As BrETS is highly expressed in the mammary gland (where intense extracellular remodelling occurs), and has been localised to the epithelial tissue, it may play a role in the regulation of matrix metalloproteinases or their inhibitors.

Genomic DNA spanning the BrETS locus was isolated in this study. It was found to encompass approximately 3kb of upstream sequence, the first two exons and intron 1. Multiple transcription start sites have been identified, indicative of a TATA-less, tissue- specific gene. Deletion constructs of the proximal promoter (850bp) were examined in cell culture. The mammary gland derived cell lines HC11 and Comma1D were used, in conjunction with a chloramphenicol acetyltransferase reporter system. A region of sequence between -70 and -200bp was identified as having up-regulated transcriptional activity above an empty expression vector in cell culture. Analysis of more distal promoter constructs resulted in decreased activity, to a level representative of basal activity. The first intron (6kb) was also examined in vitro as a complete construct and as smaller fragments. Low levels of reporter activity were seen with these constructs in cell culture. DNasel hypersensitive analysis on the immediate promoter region revealed the presence of a hypersensitive site within the first intron. This implies a regulatory element of potential importance is located here, though further research is required to determine the element(s) and significance.

The identification of regulatory regions may allow for the determination of tissue-specific elements and regions that control the expression of the gene. This information may be later extrapolated and manipulated in vivo as a means of controlling the expression of the BrETS gene.

text: A promoter characterisation of the Ets transcription factor gene, Ese-2/Elf-5 E26 transformation specific (Ets) proteins are a versatile class of transcription factors. They act both up and downstream of signal transduction cascades, are causative in cancer generation and are able to control the expression of other transcription factors. Thus they play an important role in cellular growth, proliferation and differentiation. The ets gene family is characterised by an 85 amino acid DNA binding domain and currently consists of around 30 members. They have been shown to be proto-oncogenes and it is this feature that has elicited much research into their regulation and function in both normal and neoplasic tissue. The high level of conservation within the ETS domain of the family identifies all Ets proteins as sequence-specific DNA-binding proteins. Many have been identified as having a role in the regulation of gene expression. The retention of many ets genes throughout evolution implies there would be few redundant genes (Thomas, 1993) and therefore each gene would have a unique biological role. Because of the common DNA binding domain, the regulation of the ets genes must be complex to provide specificity of action throughout the family (Laudet et al., 1999).

A cDNA sequence was isolated from mammary gland tissue and characterised as belonging to the Ets family of transcription factors. The gene was designated BrETS for Breast ETS. Recent publications have described the gene as Elf-5 (Zhou et al., 1998) and ESE-2 (Oettgen et al., 1999). BrEts showed a restricted pattern of spatial expression, being strongly expressed in the mammary gland. Its expression in the mammary gland also exhibits a tightly controlled temporal pattern, which implicates it in mammary processes such as development and differentiation. As BrETS is highly expressed in the mammary gland (where intense extracellular remodelling occurs), and has been localised to the epithelial tissue, it may play a role in the regulation of matrix metalloproteinases or their inhibitors.

Genomic DNA spanning the BrETS locus was isolated in this study. It was found to encompass approximately 3kb of upstream sequence, the first two exons and intron 1. Multiple transcription start sites have been identified, indicative of a TATA-less, tissue- specific gene. Deletion constructs of the proximal promoter (850bp) were examined in cell culture. The mammary gland derived cell lines HC11 and Comma1D were used, in conjunction with a chloramphenicol acetyltransferase reporter system. A region of sequence between -70 and -200bp was identified as having up-regulated transcriptional activity above an empty expression vector in cell culture. Analysis of more distal promoter constructs resulted in decreased activity, to a level representative of basal activity. The first intron (6kb) was also examined in vitro as a complete construct and as smaller fragments. Low levels of reporter activity were seen with these constructs in cell culture. DNasel hypersensitive analysis on the immediate promoter region revealed the presence of a hypersensitive site within the first intron. This implies a regulatory element of potential importance is located here, though further research is required to determine the element(s) and significance.

The identification of regulatory regions may allow for the determination of tissue-specific elements and regions that control the expression of the gene. This information may be later extrapolated and manipulated in vivo as a means of controlling the expression of the BrETS gene.

year: 2002

title: Time-load: Visualization of energy consumption loads over time

authors: Masoodian, Masood

abstract: It has been suggested that providing feedback allows people to better understand their energy consumption behaviour and take the necessary actions to reduce energy consumption. Here, we present the time-load visualization, which shows percentages of energy usage by different categories of devices that contribute to the total energy consumption load over time. Time-load aims to assist users with visualization of variations in energy consumption by different categories over time, and their respective, and often very unbalanced, contributions to the total energy usage load.

text: Time-load: Visualization of energy consumption loads over time It has been suggested that providing feedback allows people to better understand their energy consumption behaviour and take the necessary actions to reduce energy consumption. Here, we present the time-load visualization, which shows percentages of energy usage by different categories of devices that contribute to the total energy consumption load over time. Time-load aims to assist users with visualization of variations in energy consumption by different categories over time, and their respective, and often very unbalanced, contributions to the total energy usage load.

year: 2016

title: An investigation into the commercial feasibility of Jasus edwardsii aquaculture in New Zealand

authors: Ruru, Ian Hector

subjects: New Zealand

abstract: In 1996 New Zealand introduced legislation to allow Jasus edwardsii pueruli to be collected as "seedstock" and ongrown in a commercial aquaculture trial. The aim of this thesis was to investigate the commercial feasibility of J. edwardsii aquaculture in New Zealand by working with an organisation involved in the trial.
The methodology involved a combination of experiments in the field, in laboratories, and within an ongrowing facility. This thesis examined five key areas and found the following:
1. Data obtained in the research indicates that New Zealand will continue to rely on harvesting pueruli from the wild since significant technical obstacles remain in the development of commercial scale pueruli hatcheries. Forecast data, obtained using the delphi technique and information about current research on larval rearing, indicates that commercial scale supply of pueruli from hatcheries may occur between 2017 and 2021.
2. Harvesting trials collected a low number of pueruli with the lowest unit cost for collection being $1.80. A greater collection rate could be achieved with a better understanding of seasonal and local settlement patterns.
3. Survival rates in pueruli transporting experiments were significantly affected by stocking density, time and temperature. The safest transit conditions were achieved with 15 pueruli per litre for less than 6 hours at 14°C.
4. A range of stocking densities, from 3 lobsters per tank to 12 per tank, were tested for their effect on growth of six-month old lobsters. The lobsters were reared in a total recirculation seawater system at a constant 16°C, and fed fresh mussels (Perna canaliculus). For the 121 day trial, no statistically significant effect was detected. However, there was a trend for growth rates to be highest at 3 and 6 lobsters per tank and for growth rates at stocking densities of 9 and 12 lobsters per tank to decrease after Day 50. Lobsters showed increased mortality at the highest stocking rate.
5. A bioeconomic model of a hypothetical farm was developed to assess economic benefits and risks and to determine overall profitability. The model showed an annual net cash position of $287,100 and cumulative cash position of $859,200 by the 10th year of operation based on annual sales of 10,837 kg at $65 per kg. The model indicates that the highest cost components in the first 10 years are processing and transport, and labour. Based on the model and the scenarios examined, profitability is very sensitive to factors such as the farm size, processing and transport costs, and sale price. Biological factors such as growth rates, survival, and feed requirements also influence overall profitability. Good returns will depend upon consistent exports of high quality lobsters.
The study concludes that commercial feasibility will be affected by these key areas. The potential of J. edwardsii aquaculture in New Zealand will continue to benefit from further research.

text: An investigation into the commercial feasibility of Jasus edwardsii aquaculture in New Zealand In 1996 New Zealand introduced legislation to allow Jasus edwardsii pueruli to be collected as "seedstock" and ongrown in a commercial aquaculture trial. The aim of this thesis was to investigate the commercial feasibility of J. edwardsii aquaculture in New Zealand by working with an organisation involved in the trial.
The methodology involved a combination of experiments in the field, in laboratories, and within an ongrowing facility. This thesis examined five key areas and found the following:
1. Data obtained in the research indicates that New Zealand will continue to rely on harvesting pueruli from the wild since significant technical obstacles remain in the development of commercial scale pueruli hatcheries. Forecast data, obtained using the delphi technique and information about current research on larval rearing, indicates that commercial scale supply of pueruli from hatcheries may occur between 2017 and 2021.
2. Harvesting trials collected a low number of pueruli with the lowest unit cost for collection being $1.80. A greater collection rate could be achieved with a better understanding of seasonal and local settlement patterns.
3. Survival rates in pueruli transporting experiments were significantly affected by stocking density, time and temperature. The safest transit conditions were achieved with 15 pueruli per litre for less than 6 hours at 14°C.
4. A range of stocking densities, from 3 lobsters per tank to 12 per tank, were tested for their effect on growth of six-month old lobsters. The lobsters were reared in a total recirculation seawater system at a constant 16°C, and fed fresh mussels (Perna canaliculus). For the 121 day trial, no statistically significant effect was detected. However, there was a trend for growth rates to be highest at 3 and 6 lobsters per tank and for growth rates at stocking densities of 9 and 12 lobsters per tank to decrease after Day 50. Lobsters showed increased mortality at the highest stocking rate.
5. A bioeconomic model of a hypothetical farm was developed to assess economic benefits and risks and to determine overall profitability. The model showed an annual net cash position of $287,100 and cumulative cash position of $859,200 by the 10th year of operation based on annual sales of 10,837 kg at $65 per kg. The model indicates that the highest cost components in the first 10 years are processing and transport, and labour. Based on the model and the scenarios examined, profitability is very sensitive to factors such as the farm size, processing and transport costs, and sale price. Biological factors such as growth rates, survival, and feed requirements also influence overall profitability. Good returns will depend upon consistent exports of high quality lobsters.
The study concludes that commercial feasibility will be affected by these key areas. The potential of J. edwardsii aquaculture in New Zealand will continue to benefit from further research.

year: 2000