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: Exclusion and inaction: Academic precariat experiences of union representation in Aotearoa New Zealand

authors: Salter, Leon

abstract: During the last 40 years, neoliberal reforms to the tertiary sector have led to the casualisation of academic labour and the emergence of an academic precariat in Aotearoa New Zealand. Despite the increasing size of the academic precariat, it does not appear that their voices, concerns, or interests have been adequately represented by the national tertiary union. By drawing on open-text responses from the Precarious Academic Work Survey (PAWS) about what unions could do to improve precarious academic working conditions, we discuss the issues created by the under-representation of precarious academics by the sector union. We communicate the results via four key themes of exclusion, participation, voice, and organising. Most participants articulated frustration and disaffection with the union, suggesting the need for a shift in strategy. This study adds to the growing body of employment relations research recognising that employee voices are multiple, diverse, and fragmented; indicating that unions must attend to the differential experiences of people working in the tertiary sector attributable to employment practices.

text: Exclusion and inaction: Academic precariat experiences of union representation in Aotearoa New Zealand During the last 40 years, neoliberal reforms to the tertiary sector have led to the casualisation of academic labour and the emergence of an academic precariat in Aotearoa New Zealand. Despite the increasing size of the academic precariat, it does not appear that their voices, concerns, or interests have been adequately represented by the national tertiary union. By drawing on open-text responses from the Precarious Academic Work Survey (PAWS) about what unions could do to improve precarious academic working conditions, we discuss the issues created by the under-representation of precarious academics by the sector union. We communicate the results via four key themes of exclusion, participation, voice, and organising. Most participants articulated frustration and disaffection with the union, suggesting the need for a shift in strategy. This study adds to the growing body of employment relations research recognising that employee voices are multiple, diverse, and fragmented; indicating that unions must attend to the differential experiences of people working in the tertiary sector attributable to employment practices.

year: 2023-10-19

title: Mental health inpatient services: Improving our understanding of the needs of Maori when acutely unwell

authors: Wharewera-Mika, Julie

subjects: Maori

abstract: There are many possible explanations for the pattern of Maori overrepresentation in mental health acute services. This research project focuses on how services can optimally meet the needs of Maori to improve outcomes. This doctoral research in progress is about claiming space for Maori to have a voice in identifying factors that contribute to recovery and Whanau Ora, and offering recommendations to improve the effectiveness of existing services to better meet the needs of Maori Tangata Whaiora and Whanau.

text: Mental health inpatient services: Improving our understanding of the needs of Maori when acutely unwell There are many possible explanations for the pattern of Maori overrepresentation in mental health acute services. This research project focuses on how services can optimally meet the needs of Maori to improve outcomes. This doctoral research in progress is about claiming space for Maori to have a voice in identifying factors that contribute to recovery and Whanau Ora, and offering recommendations to improve the effectiveness of existing services to better meet the needs of Maori Tangata Whaiora and Whanau.

year: 2008