Genetic and gene expression analyses of the polycystic ovary syndrome candidate gene fibrillin-3 and other fibrillin family members in human ovaries

Several studies have demonstrated an association between polycystic ovary syndrome (PCOS) and the dinucleotide repeat microsatellite marker D19S884, which is located in intron 55 of the fibrillin-3 (FBN3) gene. Fibrillins, including FBN1 and 2, interact with latent transforming growth factor (TGF)-β-binding proteins (LTBP) and thereby control the bioactivity of TGFβs. TGFβs stimulate fibroblast replication and collagen production. The PCOS ovarian phenotype includes increased stromal collagen and expansion of the ovarian cortex, features feasibly influenced by abnormal fibrillin expression. To examine a possible role of fibrillins in PCOS, particularly FBN3, we undertook tagging and functional single nucleotide polymorphism (SNP) analysis (32 SNPs including 10 that generate non-synonymous amino acid changes) using DNA from 173 PCOS patients and 194 controls. No SNP showed a significant association with PCOS and alleles of most SNPs showed almost identical population frequencies between PCOS and control subjects. No significant differences were observed for microsatellite D19S884. In human PCO stroma/cortex (n = 4) and non-PCO ovarian stroma (n = 9), follicles (n = 3) and corpora lutea (n = 3) and in human ovarian cancer cell lines (KGN, SKOV-3, OVCAR-3, OVCAR-5), FBN1 mRNA levels were approximately 100 times greater than FBN2 and 200–1000-fold greater than FBN3. Expression of LTBP-1 mRNA was 3-fold greater than LTBP-2. We conclude that FBN3 appears to have little involvement in PCOS but cannot rule out that other markers in the region of chromosome 19p13.2 are associated with PCOS or that FBN3 expression occurs in other organs and that this may be influencing the PCOS phenotype.

Within and without family in the Icelandic sagas

The Icelandic sagas reflect a deep social interest in the nature of family obligations. Narrative tension and drama often result from carefully plotted increases in competition between families,while considerable space is given over to family biographies and genealogical information. As a result, the saga authors’ conception of the historical seems closely bound to a desire to represent family life. In Gísla saga Súrssonar and Íslendinga saga, the representation of family life extends to the situation of internal family conflicts, when the strict ethical codes underpinning the centrality of family obligations seem to be complicated and perhaps even threatened by characters’ formation of stronger bonds outside the family. The portrayal of internal family conflicts in these two sagas enabled the authors to express complex and often conflicting ethical issues.

Adolescent experiences of 'family meals' in Australia

Popular discourse laments the decline of the ‘family meal’, leading to family fragmentation and nutritional compromise. This article reports findings of a study investigating beliefs and practices surrounding the ‘family meal’, using data drawn from an on-line survey completed by 625 adolescents in Perth, Western Australia. The results challenge current concerns about the loss of the ‘family meal’, demonstrating that, for a majority, meals are eaten together rather than in isolation; are home-made rather than store bought or fast food; and are sites of conversation regardless of the presence of a television. Adolescents are divided, however, on the value of the ‘family meal’, with half seeing it as a positive experience of family togetherness and half regarding it negatively or as unimportant. The findings go some way to dispelling the notion that the ‘family meal’ no longer exists in Australia.

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.

The social orders of family mealtime

This study examined the everyday practices of families within the context of family mealtime to investigate how members accomplished mealtime interactions. Using an ethnomethodological approach, conversation analysis and membership categorization analysis, the study investigated the interactional resources that family members used to assemble their social orders moment by moment during family mealtimes. While there is interest in mealtimes within educational policy, health research and the media, there remain few studies that provide fine-grained detail about how members produce the social activity of having a family meal. Findings from this study contribute empirical understandings about families and family mealtime. Two families with children aged 2 to 10 years were observed as they accomplished their everyday mealtime activities. Data collection took place in the family homes where family members video recorded their naturally occurring mealtimes. Each family was provided with a video camera for a one-month period and they decided which mealtimes they recorded, a method that afforded participants greater agency in the data collection process and made available to the analyst a window into the unfolding of the everyday lives of the families. A total of 14 mealtimes across the two families were recorded, capturing 347 minutes of mealtime interactions. Selected episodes from the data corpus, which includes centralised breakfast and dinnertime episodes, were transcribed using the Jeffersonian system. Three data chapters examine extended sequences of family talk at mealtimes, to show the interactional resources used by members during mealtime interactions. The first data chapter explores multiparty talk to show how the uniqueness of the occasion of having a meal influences turn design. It investigates the ways in which members accomplish two-party talk within a multiparty setting, showing how one child "tells" a funny story to accomplish the drawing together of his brothers as an audience. As well, this chapter identifies the interactional resources used by the mother to cohort her children to accomplish the choralling of grace. The second data chapter draws on sequential and categorical analysis to show how members are mapped to a locally produced membership category. The chapter shows how the mapping of members into particular categories is consequential for social order; for example, aligning members who belong to the membership category "had haircuts" and keeping out those who "did not have haircuts". Additional interactional resources such as echoing, used here to refer to the use of exactly the same words, similar prosody and physical action, and increasing physical closeness, are identified as important to the unfolding talk particularly as a way of accomplishing alignment between the grandmother and grand-daughter. The third and final data analysis chapter examines topical talk during family mealtimes. It explicates how members introduce topics of talk with an orientation to their co-participant and the way in which the take up of a topic is influenced both by the sequential environment in which it is introduced and the sensitivity of the topic. Together, these three data chapters show aspects of how family members participated in family mealtimes. The study contributes four substantive themes that emerged during the analytic process and, as such, the themes reflect what the members were observed to be doing. The first theme identified how family knowledge was relevant and consequential for initiating and sustaining interaction during mealtime with, for example, members buying into the talk of other members or being requested to help out with knowledge about a shared experience. Knowledge about members and their activities was evident with the design of questions evidencing an orientation to coparticipant’s knowledge. The second theme found how members used topic as a resource for social interaction. The third theme concerned the way in which members utilised membership categories for producing and making sense of social action. The fourth theme, evident across all episodes selected for analysis, showed how children’s competence is an ongoing interactional accomplishment as they manipulated interactional resources to manage their participation in family mealtime. The way in which children initiated interactions challenges previous understandings about children’s restricted rights as conversationalists. As well as making a theoretical contribution, the study offers methodological insight by working with families as research participants. The study shows the procedures involved as the study moved from one where the researcher undertook the decisions about what to videorecord to offering this decision making to the families, who chose when and what to videorecord of their mealtime practices. Evident also are the ways in which participants orient both to the video-camera and to the absent researcher. For the duration of the mealtime the video-camera was positioned by the adults as out of bounds to the children; however, it was offered as a "treat" to view after the mealtime was recorded. While situated within family mealtimes and reporting on the experiences of two families, this study illuminates how mealtimes are not just about food and eating; they are social. The study showed the constant and complex work of establishing and maintaining social orders and the rich array of interactional resources that members draw on during family mealtimes. The family’s interactions involved members contributing to building the social orders of family mealtime. With mealtimes occurring in institutional settings involving young children, such as long day care centres and kindergartens, the findings of this study may help educators working with young children to see the rich interactional opportunities mealtimes afford children, the interactional competence that children demonstrate during mealtimes, and the important role/s that adults may assume as co-participants in interactions with children within institutional settings.

Test of a model of the effects of parental illness on youth and family functioning

Objective: Parental illness (PI) may have adverse impacts on youth and family functioning. Research in this area has suffered from the absence of a guiding comprehensive framework. This study tested a conceptual model of the effects of PI on youth and family functioning derived from the Family Ecology Framework (FEF; Pedersen & Revenson, 2005). Method. A total of 85 parents with multiple sclerosis and 127 youth completed questionnaires at Time 1 and 12 months later at Time 2. Results. Structural equation modeling results supported the FEF with regards to physical-illness disability. Specifically, the proposed mediators (role redistribution, stress, and stigma) were implicated in the processes that link parental disability to several domains of youth adjustment. The results suggest that the effects of parental depression (PD) are not mediated through these processes; rather, PD directly affects family functioning, which in turn mediates the effects onto youth adjustment. Family functioning further mediated between PD and youth well-being and behavioral-social difficulties. Conclusions. Although results support the effects of parental-illness disability on youth and family functioning via the proposed mediational mechanisms, the additive effects of PD on youth physical and mental health occur through direct and indirect (via family functioning) pathways, respectively.