Substrate specificity and structural investigation into PepO and PepW : two peptidases from Lactobacillus rhamnosus : a thesis presented in partial fulfilment of the requirements for the degree of Master of Science in Biochemistry at Massey University, Palmerston North, New Zealand

The proteolytic systems of lactic acid bacteria have important roles in the maturation and flavour development of cheese. Lactic acid bacteria pepetidases contribute to the taste of cheese through the production of low-molecular weight peptides and free amino acids. Although some lactic acid bacteria peptidases have been structurally and enzymatically characterised for their substrate specificity, there are some that are yet to be completely biochemically characterised. The aim of the present study was to investigate the substrate specificity and three-dimensional structure of two peptidases that could potentially be used as a tool to modify and control cheese bitterness and possibly other flavour attributes from Lactobacillus rhamnosus, PepO and PepW. The pepW gene was successfully cloned from L. rhamnosus into an E. coli expression system. Recombinant PepW was purified to homogeneity and was shown to exist as a hexamer of 50 kDa subunits. Recombinant PepO was expressed from a previously established L. lactis expression system and purified to homogeneity. PepO was shown to exist as a 70 kDa monomer, and function as a metallopeptidase. Pepo and PepW were shown to selectively hydrolyse chymosin-derived bovine β- and κ-casein peptides, and casein peptides extracted from Cheddar cheese. One conclusive PepO cleavage site that had not been previously characterised was identified. This was the β-casein peptide bond between Leu₆-Asn₇. Several possible PepO and PepW cleavage sites in αs₁-, β- and κ- casein were identified, suggesting that PepO has a broad endopeptidase activity, whilst PepW has a specific exopeptidase activity. Pepo and PepW crystals were successfully grown for structure determination by x-ray crystallography. Native data sets were collected for both PepO and PepW, and derivative data were collected for PepO. Structure determination was attempted using Multiple Isomorphous Replacement and Molecular Replacement techniques. Results from the substrate specificity and structural investigation of the L. rhamnosus peptidases, PepO and PepW, are presented in this thesis

The experience of midwives who work in a dual role as nurse and midwife in rural facilities in North Queensland, Australia

Background: Closure of Australian rural birthing services due to workforce shortages has serious implications for birthing women, their families, and the community. It has also had a huge impact on midwives who may find themselves in positions where they are forced to work as both a rural generalist and a midwife within the same facility and even perhaps during the same shift. This has the potential to create role conflict or role ambiguity, a known factor that impacts on job satisfaction and workforce retention. This has serious implications for maternity workforce in rural areas, already experiencing shortages. Aim: The aim of this study was to use a phenomenological approach to explore the experience of midwives who worked in a dual role of generalist nurse and midwife in a small rural hospital. Method: A purposive sampling method was utilised and midwives working in a dual role in rural facilities were invited to participate in the study. Data was collected through the use of semi-structured interviews which were then transcribed and analysed using van Manen's methodological approach. Ethics: Approval was granted by the relevant University and Health District Human Research Ethics Committee. Key findings: Three themes were identified to illustrate the experience of working in a dual role as nurse and midwife in a rural areas. These were: 1. "Because I choose to live here" (Choice/no choice) 2. "All in together, whatever the weather" (Integration) 3. "That's part of working in a small place" (Realities of rural)

Identification of novel chondroitin proteoglycans in Caenorhabditis elegans: embryonic cell division depends on CPG-1 and CPG-2.

Vertebrates produce multiple chondroitin sulfate proteoglycans that play important roles in development and tissue mechanics. In the nematode Caenorhabditis elegans, the chondroitin chains lack sulfate but nevertheless play essential roles in embryonic development and vulval morphogenesis. However, assignment of these functions to specific proteoglycans has been limited by the lack of identified core proteins. We used a combination of biochemical purification, Western blotting, and mass spectrometry to identify nine C. elegans chondroitin proteoglycan core proteins, none of which have homologues in vertebrates or other invertebrates such as Drosophila melanogaster or Hydra vulgaris. CPG-1/CEJ-1 and CPG-2 are expressed during embryonic development and bind chitin, suggesting a structural role in the egg. RNA interference (RNAi) depletion of individual CPGs had no effect on embryonic viability, but simultaneous depletion of CPG-1/CEJ-1 and CPG-2 resulted in multinucleated single-cell embryos. This embryonic lethality phenocopies RNAi depletion of the SQV-5 chondroitin synthase, suggesting that chondroitin chains on these two proteoglycans are required for cytokinesis

Teaching linguistic mimicry to improve second language pronunciation.

This thesis tests the hypothesis that a whole language approach to ESL (English As A Second Language) pronunciation with emphasis on suprasegementals through the use of linguistic mimicry is more effective than a focus on segmentals in improving native speakers perceptions of accent and comprehensibility of ESL students' pronunciation of English. The thesis is organized into seven chapters. Chapter 2 is a discussion of the factors that affect the degree of foreign accent in second language acquisition. Chapter 3 gives a background on current ESL pedagogy followed by a description of the linguistic mimicry approach used in this research in Chapter 4. Chapter 5 and 6 are discussion of Materials and Methods and Conclusions and Implications

What challenges and enablers elicit job satisfaction in rural and remote nursing in Australia: An Integrative review

Aim: To explore challenges and stressors experienced by rural and remote area nurses and identify any interventions that aided in decreasing stress and increasing job satisfaction. Background: Demand for a generalist nursing workforce in rural and remote locations exposes nurses to the same conditions as people residing there: higher mortality rates and higher incidence of chronic diseases and inadequacies in accessing health services. Design: Christmals and Gross’s integrative review framework was used with specified inclusion and exclusion criteria. Four databases were searched with no date limits. Only Australian studies were searched as international scope of practice differences for nurses could have distorted findings. Findings: Eighteen studies identified three broad themes: access to education; isolation (geographical, professional and personal) and recognition of role. Discussion: Interlinked themes showed positives and negatives from differing viewpoints. Ambivalence to education stemmed from inadequate exposure to learning and was linked with geographical isolation. Isolation was found to be less of a challenge to nurses who had an existing emotional connection with the community. Conclusion: The themes identified were recurrent and interconnecting. The benefits of working in small rural and remote communities are being used as a driver for recruitment. These benefits include higher wages, providing a sense of belonging and allowing nurses to work to their full scope and develop generalist nursing skills. The geographical isolation generates challenges through inequality in access to education and professional support, working outside their scope of practice, safety and vulnerability that comes with living remotely and adapting to extreme weather conditions

Women’s experiences of gestational breast cancer and their interactions with the health care system: a scoping review

Aim: To report the evidence of women's experiences following a diagnosis of gestational breast cancer (GBC) and their interactions with the healthcare system. Design: A systematic scoping review. Data Sources: This scoping review systematically searched Medline, CINAHL, Psych INFO, EMBASE and SCOPUS, in addition to six grey literature databases in October 2021. A 2020 PRISMA flow diagram depicting the flow of information. Review Methods: Guided by six steps in Arksey and OMalley's Framework (2005). One researcher completed the literature review, and four independently screened the titles and abstracts related to the eligibility criteria. Results: Totalling 25 articles, these studies comprise 2 quantitative, 20 qualitative, 1 mixed-method and 2 other documents, a book and debate. Thematic analysis was guided by Braun and Clarke (2006) to identify an overarching theme of adjustment that underpinned women's narratives and was reinforced by four major and several minor themes. The four major themes were: psychological impact, motherhood, treatment and communication. The relationship between the themes contextualizes the enormous complexity concerning women's experiences with GBC. Conclusion: Cancer management for GBC is complex and multifaceted. At a time of conflicting emotions for women, Multidisciplinary teams are well placed to provide support, normalize the woman's experience of motherhood, demonstrate an understanding of treatment effects, and communicate in a considerate and empathetic manner with information that is timely and relevant. GBC management involves doctors, nurses, midwives and many other healthcare professionals, which can add to the impost of diagnosis. Impact: This scoping review contributes to a better understanding of women's experience of GBC. The results may inform improvements in the support and communication for these women with GBC and their families

Integrating nursing informatics into undergraduate nursing education in Africa

This scoping review summarised the literature on the integration of NI into undergraduate nursing education in Africa

Double degree destinations: nursing or midwifery

Background: Double degrees in nursing and midwifery have evolved in Australia as a proposed solution to possible impending shortages of qualified midwives in the healthcare workforce. The double degree is seen as a more acceptable option in non-metropolitan areas in particular. Concern has been expressed however, about dilution of midwifery philosophy and graduates’ opportunities in respect of future clinical practice. Aim: This study aimed to provide a better understanding of motivations and intentions of students who undertake the Bachelor of Nursing Science/Bachelor of Midwifery double degree. Methods: A cross-sectional survey design was employed at four universities that offered double degrees in nursing and midwifery in three states of Australia. Students enrolled in first and fourth year of a double degree and graduates of a double degree were invited to complete an online survey comprised of Likert scales and items requiring free text responses. Quantitative data was analysed using SPSS and thematic analysis was used to analyse the qualitative data. Findings: Participants indicated a clear preference for midwifery as a career with this preference increasing for each cohort at each stage of study. Discussion: Primary reasons for selecting a double degree were perceived increased opportunity for employment and use of nursing skills to enhance midwifery practice in a population with growing co-morbidities. A strong identification with midwifery philosophy and identity was also demonstrated. Conclusion: An understanding of motivations and career intentions of students undertaking double degree studies can inform future program development and workforce planning

Dual degree destinations: Nursing or Midwifery?

Midwifery bodies have expressed concern that the competing ideologies of nursing science, which is closer to a medical model of care, and of midwifery can be confusing for students and counterproductive to their education as effective midwives. Proponents of dual nursing and midwifery degrees, however, argue that there is a need in rural and regional areas of Australia for graduate nurses and midwives who understand both of these ideologies and ways of working and are able to apply them both, in context, to practice in rural and regional areas. Until midwifery led model of care options become more readily accessible in rural areas, this dual nursing/midwifery qualification serves these areas well. Anecdotally, students enrolled in this dual degree appear to be focussed on careers as midwives. This study is uses a cross-sectional survey design to survey students enrolled in years one and four of two different four year dual nursing and midwifery degrees to ascertain preferences for practice area at these two stages of the course and graduates of the program at 12-36 months post-graduation to ascertain place/field of employment at this time frame. Cross sectional survey design allows data to be collected from different cohorts at varying times on the student and graduate trajectory at the same time. HREC approval is in place. Data will be downloaded and subjected to simple descriptive analyses using SPSS. Responses from each cohort will be compared to ascertain differences between proposed career trajectories of each of the student cohorts against actual positions of the graduate cohort. This study aims to provide a better understanding of students' motivations and intentions in studying a Bachelor of Nursing Science/Bachelor of Midwifery dual degree. This information is useful when considering implications for the nursing and midwifery workforces, particularly in relation to the issue of maldistribution