title: Characterisation of VMO1 in Human Tissues

authors: Crossan, Hannah Caitlin

subjects: VMO1

abstract: The Vitelline membrane outer layer protein 1 (VMO1) was first identified in
the Vitelline membrane of the chicken egg. This membrane almost entirely
protein that separates the egg white from the yolk in amniotic eggs and
acts as the last defensive barrier between pathogens and the egg yolk
with the developing embryo. Since its discovery in the vitelline membrane,
VMO1 has been isolated from a number of different tissues and animals.
The one that would appear to have the largest impact on human health
would be its discovery in the Reissner’s membrane of mice, whose
auditory system has high structural homology to humans. The Reissner’s
membrane maintains the electrolyte level in the compartments of the inner
ear as well as the resting electrochemical potential which are both critical
for correct hearing function.
The main aims of the research undertaken in this thesis was to 1)
determine the tissues in which VMO1 is or is not expressed in humans
and 2) create a genetically modified expression clone of human VMO1 to
use in further downstream protein-protein interaction studies. Our
hypothesis is that we can amplify VMO1 from human cell lines based on
available bioinformatics data.
The first objective was to determine the gene expression of VMO1 in
multiple human cell lines. RNA was extracted from three commercial cell
lines (list) and the integrity of RNA was confirmed by agarose gel
electrophoresis by the observation of two rRNA bands. From this cDNA
Abstract
iii
was made, and PCR analysis performed to identify the expression, or not, of VMO1 in these tissues. Successful amplification of VMO1 from lung cells (A549) lead to the creation of a human VMO1 expression clone, by transforming competent Escherichia coli cells with ligated VMO-1 insert/pPLUG vector. White colonies were selected for DNA extraction and confirmed as positive using colony PCR and agarose gel electrophoresis. DNA seuqencing of the positive clone confirmed the nucleotide sequence as VMO1.
The second objective was to validate the commercial antibody for human VMO1 using physiological and immunohistochemical methodology. The immunohistochemistry data suggests that the VMO1 protein is a secreted protein since signal was detected in the P5 mouse inner ear and mouse adult lung. However, multiple bands were observed in the western blot. Futher investigation is required to validate the VMO1 antibody to demonstrate that it is indeed specific to recognising its target epitope. The production of recombinant VMO1 protein would be benefical to address this question.

text: Characterisation of VMO1 in Human Tissues The Vitelline membrane outer layer protein 1 (VMO1) was first identified in
the Vitelline membrane of the chicken egg. This membrane almost entirely
protein that separates the egg white from the yolk in amniotic eggs and
acts as the last defensive barrier between pathogens and the egg yolk
with the developing embryo. Since its discovery in the vitelline membrane,
VMO1 has been isolated from a number of different tissues and animals.
The one that would appear to have the largest impact on human health
would be its discovery in the Reissner’s membrane of mice, whose
auditory system has high structural homology to humans. The Reissner’s
membrane maintains the electrolyte level in the compartments of the inner
ear as well as the resting electrochemical potential which are both critical
for correct hearing function.
The main aims of the research undertaken in this thesis was to 1)
determine the tissues in which VMO1 is or is not expressed in humans
and 2) create a genetically modified expression clone of human VMO1 to
use in further downstream protein-protein interaction studies. Our
hypothesis is that we can amplify VMO1 from human cell lines based on
available bioinformatics data.
The first objective was to determine the gene expression of VMO1 in
multiple human cell lines. RNA was extracted from three commercial cell
lines (list) and the integrity of RNA was confirmed by agarose gel
electrophoresis by the observation of two rRNA bands. From this cDNA
Abstract
iii
was made, and PCR analysis performed to identify the expression, or not, of VMO1 in these tissues. Successful amplification of VMO1 from lung cells (A549) lead to the creation of a human VMO1 expression clone, by transforming competent Escherichia coli cells with ligated VMO-1 insert/pPLUG vector. White colonies were selected for DNA extraction and confirmed as positive using colony PCR and agarose gel electrophoresis. DNA seuqencing of the positive clone confirmed the nucleotide sequence as VMO1.
The second objective was to validate the commercial antibody for human VMO1 using physiological and immunohistochemical methodology. The immunohistochemistry data suggests that the VMO1 protein is a secreted protein since signal was detected in the P5 mouse inner ear and mouse adult lung. However, multiple bands were observed in the western blot. Futher investigation is required to validate the VMO1 antibody to demonstrate that it is indeed specific to recognising its target epitope. The production of recombinant VMO1 protein would be benefical to address this question.

year: 2014

title: Election programmes in New Zealand politics: 1911-1996

authors: Gibbons, Matthew

abstract: This thesis examines the policies New Zealand political parties have emphasised in their key policy statements, the ideological dimensions of party competition, how the political agenda has changed over time, and whether parties have acted in ways compatible with responsible party government and pluralist theories of democracy. These central political debates are studied using all the election programmes issued by the main New Zealand political parties between 1911 and 1996.

Content analysis results for the 1911-1996 period reveal enduring differences between left and right-wing parties on economic, foreign policy and some conservative themes. While some ‘new politics’ issues such as the environment and culture have become much more important over time, the data confirms that other new politics issues such as freedom and democracy have been on New Zealand’s political agenda for most of the twentieth century. 

When charted on a left-right dimension, parties’ positions were largely as expected. The main left and right-wing parties have only crossed over once, and then only fractionally on the economic policy dimension. Contrary to theoretical expectations right-wing parties have often taken very moderate positions on the new politics-old politics dimension, although left-wing parties have usually been more supportive of new politics policies than right-wing parties. Differences between Labour and the main right-wing parties have usually been larger on economic than on new politics issues. However, between 1972 and 1990 differences between Labour and National on new politics issues were much larger than on economic themes.

In their programmes parties have usually campaigned in a manner encouraging rational voting, and have taken relatively similar positions on potentially divisive ethnic policy issues. There is one obvious case of agenda setting: the main parties have kept the most morally contentious issues out of party politics. The enduring policy differences between the main parties will have made it easier for less well informed voters to vote in a rational manner. Party movement results indicate that only during the 1980s were National’s and Labour’s manifestos a poor guide to their subsequent policy direction. 

Policy implementation results for New Zealand using Klingemann, Hofferbert and Budge’s methodology showed high implementation rates by international standards over the 1946-1994 period. Often the manifestos of opposition parties were positively associated with government expenditure trends. This shows that parties’ commitment to mandate theory has been qualified by their support for pluralist norms of interest group consultation. The results indicated that between the early 1980s and 1994 the relationship between manifesto emphases and government expenditure for transfers and health has deteriorated. In other areas policy implementation rates remained high, indicating that politicians often still behaved in ways compatible with the principles of responsible party government. The regression equations Klingemann, Hofferbert and Budge use to calculate policy implementation were also reformulated to produce better and theoretically stronger results.

text: Election programmes in New Zealand politics: 1911-1996 This thesis examines the policies New Zealand political parties have emphasised in their key policy statements, the ideological dimensions of party competition, how the political agenda has changed over time, and whether parties have acted in ways compatible with responsible party government and pluralist theories of democracy. These central political debates are studied using all the election programmes issued by the main New Zealand political parties between 1911 and 1996.

Content analysis results for the 1911-1996 period reveal enduring differences between left and right-wing parties on economic, foreign policy and some conservative themes. While some ‘new politics’ issues such as the environment and culture have become much more important over time, the data confirms that other new politics issues such as freedom and democracy have been on New Zealand’s political agenda for most of the twentieth century. 

When charted on a left-right dimension, parties’ positions were largely as expected. The main left and right-wing parties have only crossed over once, and then only fractionally on the economic policy dimension. Contrary to theoretical expectations right-wing parties have often taken very moderate positions on the new politics-old politics dimension, although left-wing parties have usually been more supportive of new politics policies than right-wing parties. Differences between Labour and the main right-wing parties have usually been larger on economic than on new politics issues. However, between 1972 and 1990 differences between Labour and National on new politics issues were much larger than on economic themes.

In their programmes parties have usually campaigned in a manner encouraging rational voting, and have taken relatively similar positions on potentially divisive ethnic policy issues. There is one obvious case of agenda setting: the main parties have kept the most morally contentious issues out of party politics. The enduring policy differences between the main parties will have made it easier for less well informed voters to vote in a rational manner. Party movement results indicate that only during the 1980s were National’s and Labour’s manifestos a poor guide to their subsequent policy direction. 

Policy implementation results for New Zealand using Klingemann, Hofferbert and Budge’s methodology showed high implementation rates by international standards over the 1946-1994 period. Often the manifestos of opposition parties were positively associated with government expenditure trends. This shows that parties’ commitment to mandate theory has been qualified by their support for pluralist norms of interest group consultation. The results indicated that between the early 1980s and 1994 the relationship between manifesto emphases and government expenditure for transfers and health has deteriorated. In other areas policy implementation rates remained high, indicating that politicians often still behaved in ways compatible with the principles of responsible party government. The regression equations Klingemann, Hofferbert and Budge use to calculate policy implementation were also reformulated to produce better and theoretically stronger results.

year: 2000

title: Māori & Psychology Research Unit Annual Report 2014

authors: Waitoki, Waikaremoana

abstract: Annual report of the Māori and Psychology Research Unit (MPRU) 2014. The unit was established in August of 1997. The unit is designed to provide a catalyst and support network for enhancing research concerning the psychological needs, aspirations, and priorities of Maori people. The MPRU is well situated to draw together skilled and experienced interdisciplinary research groups by networking and establishing working relationships with staff and students within the Faculty of Arts and Social Sciences, the University, and the wider community.

text: Māori & Psychology Research Unit Annual Report 2014 Annual report of the Māori and Psychology Research Unit (MPRU) 2014. The unit was established in August of 1997. The unit is designed to provide a catalyst and support network for enhancing research concerning the psychological needs, aspirations, and priorities of Maori people. The MPRU is well situated to draw together skilled and experienced interdisciplinary research groups by networking and establishing working relationships with staff and students within the Faculty of Arts and Social Sciences, the University, and the wider community.

year: 2014