Investigating relationships between relationship quality, customer loyalty and cooperation : an empirical study of convenience stores' franchise chain systems

Small business has been shown to contribute significantly to a nation’s economic development. Small business owners typically confront challenges, uncertainty, and risks while operating new businesses. Franchising has become a way to minimize the risks of small business management (Chiou et al., 2004); however, a franchise system is not a guarantee of business success (Lee and Karkovista, 2001). A poor franchising relationship between franchisors and franchisees can result in franchise failure, such as termination and closure, or franchisee exit (Frazer and Winzar, 2005).

Characterisation of the TaNF-Y family of transcription factors in wheat

Light plays a unique role for plants as it is both a source of energy for growth and a signal for development. Light captured by the pigments in the light harvesting complexes is used to drive the synthesis of the chemical energy required for carbon assimilation. The light perceived by photoreceptors activates effectors, such as transcription factors (TFs), which modulate the expression of light-responsive genes. Recently, it has been speculated that increasing the photosynthetic rate could further improve the yield potential of three carbon (C3) crops such as wheat. However, little is currently known about the transcriptional regulation of photosynthesis genes, particularly in crop species. Nuclear factor Y (NF-Y) TF is a functionally diverse regulator of growth and development in the model plant species, with demonstrated roles in embryo development, stress response, flowering time and chloroplast biogenesis. Furthermore, a light-responsive NF-Y binding site (CCAAT-box) is present in the promoter of a spinach photosynthesis gene. As photosynthesis genes are co-regulated by light and co-regulated genes typically have similar regulatory elements in their promoters, it seems likely that other photosynthesis genes would also have light-responsive CCAAT-boxes. This provided the impetus to investigate the NF-Y TF in bread wheat. This thesis is focussed on wheat NF-Y members that have roles in light-mediated gene regulation with an emphasis on their involvement in the regulation of photosynthesis genes. NF-Y is a heterotrimeric complex, comprised of the three subunits NF-YA, NF-YB and NF-YC. Unlike the mammalian and yeast counterparts, each of the three subunits is encoded by multiple genes in Arabidopsis. The initial step taken in this study was the identification of the wheat NF-Y family (Chapter 3). A search of the current wheat nucleotide sequence databases identified 37 NF-Y genes (10 NF-YA, 11 NF-YB, 14 NF-YC & 2 Dr1). Phylogenetic analysis revealed that each of the three wheat NF-Y (TaNF-Y) subunit families could be divided into 4-5 clades based on their conserved core regions. Outside of the core regions, eleven motifs were identified to be conserved between Arabidopsis, rice and wheat NF-Y subunit members. The expression profiles of TaNF-Y genes were constructed using quantitative real-time polymerase chain reaction (RT-PCR). Some TaNF-Y subunit members had little variation in their transcript levels among the organs, while others displayed organ-predominant expression profiles, including those expressed mainly in the photosynthetic organs. To investigate their potential role in light-mediated gene regulation, the light responsiveness of the TaNF-Y genes were examined (Chapters 4 and 5). Two TaNF-YB and five TaNF-YC members were markedly upregulated by light in both the wheat leaves and seedling shoots. To identify the potential target genes of the light-upregulated NF-Y subunit members, a gene expression correlation analysis was conducted using publically available Affymetrix Wheat Genome Array datasets. This analysis revealed that the transcript expression levels of TaNF-YB3 and TaNF-YC11 were significantly correlated with those of photosynthesis genes. These correlated express profiles were also observed in the quantitative RT-PCR dataset from wheat plants grown under light and dark conditions. Sequence analysis of the promoters of these wheat photosynthesis genes revealed that they were enriched with potential NF-Y binding sites (CCAAT-box). The potential role of TaNF-YB3 in the regulation of photosynthetic genes was further investigated using a transgenic approach (Chapter 5). Transgenic wheat lines constitutively expressing TaNF-YB3 were found to have significantly increased expression levels of photosynthesis genes, including those encoding light harvesting chlorophyll a/b-binding proteins, photosystem I reaction centre subunits, a chloroplast ATP synthase subunit and glutamyl-tRNA reductase (GluTR). GluTR is a rate-limiting enzyme in the chlorophyll biosynthesis pathway. In association with the increased expression of the photosynthesis genes, the transgenic lines had a higher leaf chlorophyll content, increased photosynthetic rate and had a more rapid early growth rate compared to the wild-type wheat. In addition to its role in the regulation of photosynthesis genes, TaNF-YB3 overexpression lines flower on average 2-days earlier than the wild-type (Chapter 6). Quantitative RT-PCR analysis showed that there was a 13-fold increase in the expression level of the floral integrator, TaFT. The transcript levels of other downstream genes (TaFT2 and TaVRN1) were also increased in the transgenic lines. Furthermore, the transcript levels of TaNF-YB3 were significantly correlated with those of constans (CO), constans-like (COL) and timing of chlorophyll a/b-binding (CAB) expression 1 [TOC1; (CCT)] domain-containing proteins known to be involved in the regulation of flowering time. To summarise the key findings of this study, 37 NF-Y genes were identified in the crop species wheat. An in depth analysis of TaNF-Y gene expression profiles revealed that the potential role of some light-upregulated members was in the regulation of photosynthetic genes. The involvement of TaNF-YB3 in the regulation of photosynthesis genes was supported by data obtained from transgenic wheat lines with increased constitutive expression of TaNF-YB3. The overexpression of TaNF-YB3 in the transgenic lines revealed this NF-YB member is also involved in the fine-tuning of flowering time. These data suggest that the NF-Y TF plays an important role in light-mediated gene regulation in wheat.

Free convection heat transfer through coupled thermal boundary layers in a partitioned triangular enclosure

A numerical investigation has been carried out for the coupled thermal boundary layers on both sides of a partition placed in an isosceles triangular enclosure along its middle symmetric line. The working fluid is considered as air which is initially quiescent. A sudden temperature difference between two zones of the enclosure has been imposed to trigger the natural convection. It is anticipated from the numerical simulations that the coupled thermal boundary layers development adjacent to the partition undergoes three distinct stages; namely an initial stage, a transitional stage and a steady state stage. Time dependent features of the coupled thermal boundary layers as well as the overall natural convection flow in the partitioned enclosure have been discussed and compared with the non-partitioned enclosure. Moreover, heat transfer as a form of local and overall average Nusselt number through the coupled thermal boundary layers and the inclined walls is also examined. The details results will be discussed in the full paper.

Communications and learning structures that facilitate transfer of knowledge at innovation transition points

Innovation processes are rarely smooth and disruptions often occur at transition points were one knowledge domain passes the technology on to another domain. At these transition points communication is a key component in assisting the smooth hand over of technologies. However for smooth transitions to occur we argue that appropriate structures have to be in place and boundary spanning activities need to be facilitated. This paper presents three case studies of innovation processes and the findings support the view that structures and boundary spanning are essential for smooth transitions. We have explained the need to pass primary responsibility between agents to successfully bring an innovation to market. We have also shown the need to combine knowledge through effective communication so that absorptive capacity is built in process throughout the organisation rather than in one or two key individuals.

The inherent challenges of knowledge transfer across countries and cultures

Purpose — The purpose of this chapter is to outline the findings of research into knowledge transfer across countries and culture and the implications of the findings for human resource practitioners. Methodology/approach — Use of a qualitative case study methodology approach was employed. Findings — The findings highlight the inherent difficulty of knowledge transfer and the broad range of factors that influence the knowledge transfer process and which are connected together in a complex and non-linear manner. Research implications — The research revealed insight into those managers most likely to be effective at knowledge transfer and what characteristics and experiences had supported this openness to knowledge from outside of their cultural context. The lessons from the research can be incorporated into the selection and development processes for expatriates. Originality value — The study affirmed the critical importance of face-to-face interaction in knowledge transfer. The research also resulted in the development of a practical model and tool, which pulls together the broad range of factors that impact on knowledge transfer.

Can transfer of learning be enriched by valuing knowledge transformation and co-creation? Exploring new ways to improve training design

This paper argues that a possible cause of issues with management education outcomes is the fact that most training models operate from a limited ‘transfer’ metaphor. This theoretical paper contends that by reconceptualising existing models, specifically Holton’s transfer of learning model, to incorporate multiple processes and acknowledge the importance of educator- or trainer-student interaction in co-creating knowledge, there is potential to improve training design and ultimately achieve more satisfactory training outcomes.

Arc voltage behavior in GMAW-P under different drop transfer modes

Purpose: Experimental measurements have been made to investigate meaning of the change in voltage for the pulse gas metal arc welding (GMAW-P) process operating under different drop transfer modes. Design/methodology/approach: Welding experiments with different values of pulsing parameter and simultaneous recording of high speed camera pictures and welding signals (such as current and voltage) were used to identify different drop transfer modes in GMAW-P. The investigation is based on the synchronization of welding signals and high speed camera to study the behaviour of voltage signal under different drop transfer modes. Findings: The results reveal that the welding arc is significantly affected by the molten droplet detachment. In fact, results indicate that sudden increase and drop in voltage just before and after the drop detachment can be used to characterize the voltage behaviour of different drop transfer mode in GMAW-P. Research limitations/implications: The results show that voltage signal carry rich information about different drop transfer occurring in GMAW-P. Hence it’s possible to detect different drop transfer modes. Future work should concentrate on development of filters for detection of different drop transfer modes. Originality/value: Determination of drop transfer mode with GMAW-P is crucial for the appropriate selection of pulse welding parameters. As change in drop transfer mode results in poor weld quality in GMAW-P, so in order to estimate the working parameters and ensure stable GMAW-P understanding the voltage behaviour of different drop transfer modes in GMAW-P will be useful. However, in case of GMAW-P hardly any attempt is made to analyse the behaviour of voltage signal for different drop transfer modes. This paper analyses the voltage signal behaviour of different drop transfer modes for GMAW-P.

Technology transfer evaluation in the high technology industry : an interdisciplinary perspective

The underlying objective of this study was to develop a novel approach to evaluate the potential for commercialisation of a new technology. More specifically, this study examined the 'ex-ante'. evaluation of the technology transfer process. For this purpose, a technology originating from the high technology sector was used. The technology relates to the application of software for the detection of weak signals from space, which is an established method of signal processing in the field of radio astronomy. This technology has the potential to be used in commercial and industrial areas other than astronomy, such as detecting water leakages in pipes. Its applicability to detecting water leakage was chosen owing to several problems with detection in the industry as well as the impact it can have on saving water in the environment. This study, therefore, will demonstrate the importance of interdisciplinary technology transfer. The study employed both technical and business evaluation methods including laboratory experiments and the Delphi technique to address the research questions. There are several findings from this study. Firstly, scientific experiments were conducted and these resulted in a proof of concept stage of the chosen technology. Secondly, validation as well as refinement of criteria from literature that can be used for „ex-ante. evaluation of technology transfer has been undertaken. Additionally, after testing the chosen technology.s overall transfer potential using the modified set of criteria, it was found that the technology is still in its early stages and will require further development for it to be commercialised. Furthermore, a final evaluation framework was developed encompassing all the criteria found to be important. This framework can help in assessing the overall readiness of the technology for transfer as well as in recommending a viable mechanism for commercialisation. On the whole, the commercial potential of the chosen technology was tested through expert opinion, thereby focusing on the impact of a new technology and the feasibility of alternate applications and potential future applications.

Life Raft Australia?

In the helter skelter of people legally and illegally seeking refuge in Australia, often overlooked are the realities of how to obtain that hoped for better life. One of the avenues chosen to achieve their dream is higher education; however, once enrolled in higher education many such students [termed Culturally and Linguistically Diverse (CALD)] find they are ill equipped to achieve academic success; they become overwhelmed by the pressures of study/family/work; and ultimately lose heart and their place at university. Life raft Australia has just become another disillusioning vehicle to a depressing future. This paper discusses how universities across Australia are now putting processes in place that will enable CALD students to confront and overcome academic and social barriers, and thus realise their dream of a better life. Exemplars from the University of Tasmania provide practical models of how higher education institutions, not only in Australia but globally can assist immigrant students to self actualize; to be empowered to become integral and successful members of the Australian community.

Life Raft Australia?

In the helter skelter of people legally and illegally seeking refuge in Australia, often overlooked are the realities of how to obtain that hoped for better life. One of the avenues chosen to achieve their dream is higher education; however, once enrolled in higher education many such students [termed Culturally and Linguistically Diverse (CALD)] find they are ill equipped to achieve academic success; they become overwhelmed by the pressures of study/family/work; and ultimately lose heart and their place at university. Life raft Australia has just become another disillusioning vehicle to a depressing future. This paper discusses how universities across Australia are now putting processes in place that will enable CALD students to confront and overcome academic and social barriers, and thus realise their dream of a better life. Exemplars from the University of Tasmania provide practical models of how higher education institutions, not only in Australia but globally can assist immigrant students to self actualize; to be empowered to become integral and successful members of the Australian community.

rac-4-Carbamoylpiperidinium cis-2-carboxycyclohexane-1-carboxylate

In the title salt, racemic C6H12N2O+ C8H11O4- from the reaction of cis-cyclohexane-1,2-dicarboxylic anhydride with isonipecotamide, the cations are linked into duplex chain substructures through both centrosymmetric cyclic head-to-head 'amide motif' hydrogen-bonding associations [graph set R2/2(8)] and 'side-by-side' R2/2(14) associations. The anions are incorporated into the chains through cyclic R3/4(10) interactions involving amide and piperidinium N-H...O(carboxyl) hydrogen bonds which, together with inter-anion carboxylic acid O-H...O(carboxyl) hydrogen bonds, give a two-dimensional layered structure extending along (011).

The role of insulin and IGF2 signalling on metabolic pathways in prostate cancer progression

Prostate cancer (CaP) is the most commonly diagnosed cancer in males in Australia, North America, and Europe. If found early and locally confined, CaP can be treated with radical prostatectomy or radiation therapy; however, 25-40% patients will relapse and go on to advanced disease. The most common therapy in these cases is androgen deprivation therapy (ADT), which suppresses androgen production from the testis. Lack of the testicular androgen supply causes cells of the prostate to undergo apoptosis. However, in some cases the regression initially seen with ADT eventually gives way to a growth of a population of cancerous cells that no longer require testicular androgens. This phenotype is essentially fatal and is termed castrate resistant prostate cancer (CRPC). In addition to eventual regression, there are many undesirable side effects which accompany ADT, including development of a metabolic syndrome, which is defined by the U.S. National Library of Medicine as “a combination of medical disorders that increase the risk of developing cardiovascular disease and diabetes.” This project will focus on the effect of ADT induced hyperinsulinemia, as mimicked by treating androgen receptor positive CaP cells with insulin in a serum (hormone) deprived environment. While this side effect is not widely explored, in this thesis it is demonstrated for the first time that insulin upregulates pathways important to CaP progression. Our group has previously shown that during CaP progression, the enzymes necessary for de novo steroidogenesis are upregulated in the LNCaP xenograft model, total steroid levels are increased in tumours compared to pre castrate levels, and de novo steroidogenesis from radio-labelled acetate has been demonstrated. Because of the CaP dependence on AR for survival, we and other groups believe that CaP cells carry out de novo steroidogenesis to survive in androgen deprived conditions. Because (a) men on ADT often develop metabolic syndrome, and (b) men with lifestyle-induced obesity and hyperinsulinemia have worse prognosis and faster disease progression, and because (c) insulin causes steroidogenesis in other cell lines, the hypothesis that insulin may contribute to CaP progression through upregulation of steroidogenesis was explored. Insulin upregulates steroidogenesis enzymes at the mRNA level in three AR positive cell lines, as well as upregulating these enzymes at the protein level in two cell lines. It has also been demonstrated that insulin increases mitochondrial (functional) levels of steroid acute regulatory protein (StAR). Furthermore, insulin causes increased levels of total steroids in and induction of de novo steroid synthesis by insulin has been demonstrated at levels induced sufficient to activate AR. The effect of insulin analogs on CaP steroidogenesis in LNCaP and VCaP cells has also been investigated because epidemiological studies suggest that some of the analogs developed may have more cancer stimulatory effects than normal insulin. In this project, despite the signalling differences between glargine, X10, and insulin, these analogs did not appear to induce steroidogenesis any more potently that normal insulin. The effect of insulin of MCF7breast cancer cells was also investigated with results suggesting that breast cancer cells may be capable of de novo steroidogenesis, and that increase in estradiol production may be exacerbated by insulin. Insulin has also been long known to stimulate lipogenesis in the liver and adipocytes, and has been demonstrated to increase lipogenesis in breast cancer cells; therefore, investigation of the effect of insulin on lipogenesis, which is a hallmark of aggressive cancers, was investigated. In CaP progression sterol regulatory element binding protein (SREBP) is dysregulated and upregulates fatty acid synthase (FASN), acetyl CoA-carboxylase, and other lipogenesis genes. SREBP is important for steroidogenesis and in this project has been shown to be upregulated by insulin in CaP cells. Fatty acid synthesis provides building blocks of membrane growth, provides substrates for acid oxidation, the main energy source for CaP cells, provides building blocks for anti-apoptotic and proinflammatory molecules, and provides molecules that stimulate steroidogenesis. In this project it has been shown that insulin upregulates FASN and ACC, which synthesize fatty acids, as well as upregulating hormone sensitive lipase (HSL), diazepam-binding inhibitor (DBI), and long-chain acyl-CoA synthetase 3 (ACSL3), which contribute to lipid activation of steroidogenesis. Insulin also upregulates total lipid levels and de novo lipogenesis, which can be suppressed by inhibition of the insulin receptor (INSR). The fatty acids synthesized after insulin treatment are those that have been associated with CaP; furthermore, microarray data suggests insulin may upregulate fatty acid biosynthesis, metabolism and arachidonic acid metabolism pathways, which have been implicated in CaP growth and survival. Pharmacological agents used to treat patients with hyperinsulinemia/ hyperlipidemia have gained much interest in regards to CaP risk and treatment; however, the scientific rationale behind these clinical applications has not been examined. This thesis explores whether the use of metformin or simvastatin would decrease either lipogenesis or steroidogenesis or both in CaP cells. Simvastatin is a 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitor, which blocks synthesis of cholesterol, the building block of steroids/ androgens. It has also been postulated to down regulate SREBP in other metabolic disorders. It has been shown in this thesis, in LNCaP cells, that simvastatin inhibited and decreased insulin induced steroidogenesis and lipogenesis, respectively, but increased these pathways in the absence of insulin. Conversely, metformin, which activates AMP-activated protein kinase (AMPK) to shut down lipogenesis, cholesterol synthesis, and protein synthesis, highly suppresses both steroidogenesis and lipogenesis in the presence and absence of insulin. Lastly, because it has been demonstrated to increase steroidogenesis in other cell lines, and because the elucidation of any factors affecting steroidogenesis is important to understanding CaP, the effect of IGF2 on steroidogenesis in CaP cells was investigated. In patient samples, as men progress to CRPC, IGF2 mRNA and the protein levels of the receptors it may signal through are upregulated. It has also been demonstrated that IGF2 upregulates steroidogenic enzymes at both the mRNA and protein levels in LNCaP cells, increases intracellular and secreted steroid/androgen levels in LNCaPs to levels sufficient to stimulate the AR, and upregulated de novo steroidogenesis in LNCaPs and VCaPs. As well, inhibition of INSR and insulin-like growth factor 1 receptor (IGF1R), which IGF2 signals through, suggests that induction of steroidogenesis may be occurring predominantly through IGF1R. In summary, this project has illuminated for the first time that insulin is likely to play a large role in cancer progression, through upregulation of the steroidogenesis and lipogenesis pathways at the mRNA and protein levels, and production levels, and demonstrates a novel role for IGF-II in CaP progression through stimulation of steroidogenesis. It has also been demonstrated that metformin and simvastatin drugs may be useful in suppressing the insulin induction of these pathways. This project affirms the pathways by which ADT- induced metabolic syndrome may exacerbate CaP progression and strongly suggests that the monitoring and modulation of the metabolic state of CaP patients could have a strong impact on their therapeutic outcomes.

Evaluation of an Australian solar community : implications for education and training

1.1 Background What is renewable energy education and training? A cursory exploration of the International Solar Energy Society website (www.ises.org) reveals numerous references to education and training, referring collectively to concepts of the transfer and exchange of information and good practices, awareness raising and skills development. The purposes of such education and training relate to changing policy, stimulating industry, improving quality control and promoting the wider use of renewable energy sources. The primary objective appears to be to accelerate a transition to a better world for everyone (ISEE), as the greater use of renewable energy is seen as key to climate recovery; world poverty alleviation; advances in energy security, access and equality; improved human and environmental health; and a stabilized society. The Solar Cities project – Habitats of Tomorrow – aims at promoting the greater use of renewable energy within the context of long term planning for sustainable urban development. The focus is on cities or communities as complete systems; each one a unique laboratory allowing for the study of urban sustainability within the context of a low carbon lifestyle. The purpose of this paper is to report on an evaluation of a Solar Community in Australia, focusing specifically on the implications (i) for our understandings and practices in renewable energy education and training and (ii) for sustainability outcomes. 1.2 Methodology The physical context is a residential Ecovillage (a Solar Community) in sub-tropical Queensland, Australia (latitude 28o south). An extensive Architectural and Landscape Code (A&LC) ‘premised on the interconnectedness of all things’ and embracing ‘both local and global concerns’ governs the design and construction of housing in the estate: all houses are constructed off-ground (i.e. on stumps or stilts) and incorporate a hybrid approach to the building envelope (mixed use of thermal mass and light-weight materials). Passive solar design, gas boosted solar water heaters and a minimum 1kWp photovoltaic system (grid connected) are all mandatory, whilst high energy use appliances such as air conditioners and clothes driers are not permitted. Eight families participated in an extended case study that encompassed both quantitative and qualitative approaches to better understand sustainable housing (perceived as a single complex technology) through its phases of design, construction and occupation. 1.3 Results The results revealed that the level of sustainability (i.e. the performance outcomes in terms of a low-carbon lifestyle) was impacted on by numerous ‘players’ in the supply chain, such as architects, engineers and subcontractors, the housing market, the developer, product manufacturers / suppliers / installers and regulators. Three key factors were complicit in the level of success: (i) systems thinking; (ii) informed decision making; and (iii) environmental ethics and business practices. 1.4 Discussion The experiences of these families bring into question our understandings and practices with regard to education and training. Whilst increasing and transferring knowledge and skills is essential, the results appear to indicate that there is a strong need for expanding our education efforts to incorporate foundational skills in complex systems and decision making processes, combined with an understanding of how our individual and collective values and beliefs impact on these systems and processes.

Microbial colonisation of human follicular fluid and adverse in vitro fertilisation outcomes

This study, investigating 263 women undergoing trans-vaginal oocyte retrieval for in vitro fertilisation (IVF) found that microorganisms colonising follicular fluid contributed to adverse IVF (pre-implantation) and pregnancy (post-implantation) outcomes including poor quality embryos, failed pregnancy and early pregnancy loss (< 37 weeks gestation). Some microorganisms also showed in vitro growth patterns in liquid media that appeared to be enhanced by the hormonal stimulation protocol used for oocyte retrieval. Elaborated cytokines within follicular fluid were also associated with adverse IVF outcomes. This study is imperative because infertility affects 16% of the human population and the numbers of couples needing assistance continues to increase. Despite significant improvements in the technical aspects of assisted reproductive technologies (ART), the live birth rate has not increased proportionally. Overt genital tract infection has been associated with both infertility and adverse pregnancy outcomes (including miscarriage and preterm birth) as a direct result of the infection or the host response to it. Importantly, once inflammation had become established, medical treatment often failed to prevent these significant adverse outcomes. Current evaluations of fertility focus on the ovary as a site of steroid hormone production and ovulation. However, infertility as a result of subclinical colonisation of the ovary has not been reported. Furthermore, identification of the microorganisms present in follicular fluid and the local cytokine profile may provide clinicians with an early indication of the prognosis for IVF treatment in infertile couples, thus allowing antimicrobial treatment and/or counselling about possible IVF failure. During an IVF cycle, multiple oocytes undergo maturation in vivo in response to hormonal hyperstimulation. Oocytes for in vitro insemination are collected trans-vaginally. The follicular fluid that bathes the maturing oocyte in vivo, usually is discarded as part of the IVF procedure, but provides a unique opportunity to investigate microbial causes of adverse IVF outcomes. Some previous studies have identified follicular fluid markers that predict IVF pregnancy outcomes. However, there have not been any detailed microbiological studies of follicular fluid. For this current study, paired follicular fluid and vaginal secretion samples were collected from women undergoing IVF cycles to determine whether microorganisms in follicular fluid were associated with adverse IVF outcomes. Microorganisms in follicular fluid were regarded as either "colonisers" or "contaminants"; colonisers, if they were unique to the follicular fluid sample, and contaminants if the same microorganisms were detected in the vaginal and follicular fluid samples indicating that the follicular fluid was merely contaminated during the oocyte retrieval process. Quite unexpectedly, by these criteria, we found that follicular fluid from approximately 30% of all subjects was colonised with bacteria. Fertile and infertile women with colonised follicular fluid had decreased embryo transfer rates and decreased pregnancy rates compared to women with contaminated follicular fluids. The observation that follicular fluid was not always sterile, but contained a diverse range of microorganisms, is novel. Many of the microorganisms we detected in follicular fluid are known opportunistic pathogens that have been detected in upper genital tract infections and are associated with adverse pregnancy outcomes. Bacteria were able to survive for at least 28 weeks in vitro, in cultures of follicular fluid. Within 10 days of establishing these in vitro cultures, several species (Lactobacillus spp., Bifidobacterium spp., Propionibacterium spp., Streptococcus spp. and Salmonella entericus) had formed biofilms. Biofilms play a major role in microbial pathogenicity and persistence. The propensity of microbial species to form biofilms in follicular fluid suggests that successful treatment of these infections with antimicrobials may be difficult. Bifidobacterium spp. grew, in liquid media, only if concentrations of oestradiol and progesterone were similar to those achieved in vivo during an IVF cycle. In contrast, the growth of Streptococcus agalactiae and Escherichia coli was inhibited or abolished by the addition of these hormones to culture medium. These data suggest that the likelihood of microorganisms colonising follicular fluid and the species of bacteria involved is influenced by the stage of the menstrual cycle and, in the case of IVF, the nature and dose of steroid hormones administered for the maturation of multiple oocytes in vivo. Our findings indicate that the elevated levels of steroid hormones during an IVF cycle may influence the microbial growth within follicular fluid, suggesting that the treatment itself will impact on the microflora present in the female upper genital tract during pre-conception and early post-conception phases of the cycle. The effect of the host immune response on colonising bacteria and on the outcomes of IVF also was investigated. White blood cells reportedly compose between 5% and 15% of the cell population in follicular fluid. The follicular membrane is semi-permeable and cells are actively recruited as part of the normal menstrual cycle and in response to microorganisms. A previous study investigated follicular fluid cytokines from infertile women and fertile oocyte donors undergoing IVF, and concluded that there were no significant differences in the cytokine concentrations between the two groups. However, other studies have reported differences in the follicular fluid cytokine levels associated with infertile women with endometriosis or polycystic ovary syndrome. In this study, elevated levels of interleukin (IL)-1 á, IL-1 â and vascular endothelial growth factor (VEGF) in vaginal fluid were associated with successful fertilisation, which may be useful marker for successful fertilisation outcomes for women trying to conceive naturally or prior to oocyte retrieval for IVF. Elevated levels of IL-6, IL-12p40, granulocyte colony stimulating factor (GCSF) and interferon-gamma (IFN ã) in follicular fluid were associated with successful embryo transfer. Elevated levels of pro-inflammatory IL-18 and decreased levels of anti-inflammatory IL-10 were identified in follicular fluid from women with idiopathic infertility. Successful fertilisation and implantation is dependent on a controlled pro-inflammatory environment, involving active recruitment of pro-inflammatory mediators to the genital tract as part of the menstrual cycle and early pregnancy. However, ongoing pregnancy requires an enhanced anti-inflammatory environment to ensure that the maternal immune system does not reject the semi-allergenic foetus. The pro-inflammatory skew in the follicular fluid of women with idiopathic infertility, correlates with normal rates of fertilisation, embryo discard and embryo transfer, observed for this cohort, which were similar to the outcomes observed for fertile women. However, their pregnancy rate was reduced compared to fertile women. An altered local immune response in follicular fluid may provide a means of explaining infertility in this cohort, previously defined as 'idiopathic'. This study has found that microorganisms colonising follicular fluid may have contributed to adverse IVF and pregnancy outcomes. Follicular fluid bathes the cumulus oocyte complex during the in vivo maturation process, and microorganisms in the fluid, their metabolic products or the local immune response to these microorganisms may result in damage to the oocytes, degradation of the cumulus or contamination of the IVF culture system. Previous studies that have discounted bacterial contamination of follicular fluid as a cause of adverse IVF outcomes failed to distinguish between bacteria that were introduced into the follicular fluid at the time of trans-vaginal oocyte retrieval and those that colonised the follicular fluid. Those bacteria that had colonised the fluid may have had time to form biofilms and to elicit a local immune response. Failure to draw this distinction has previously prevented consideration of bacterial colonisation of follicular fluid as a cause of adverse IVF outcomes. Several observations arising from this study are of significance to IVF programs. Follicular fluid is not always sterile and colonisation of follicular fluid is a cause of adverse IVF and pregnancy outcomes. Hormonal stimulation associated with IVF may influence whether follicular fluid is colonised and enhance the growth of specific species of bacteria within follicular fluid. Bacteria in follicular fluid may form biofilms and literature has reported that this may influence their susceptibility to antibiotics. Monitoring the levels of selected cytokines within vaginal secretions may inform fertilisation outcomes. This study has identified novel factors contributing to adverse IVF outcomes and that are most likely to affect also natural conception outcomes. Early intervention, possibly using antimicrobial or immunological therapies may reduce the need for ART and improve reproductive health outcomes for all women.