Intrinsics and dynamics of fat grafts : an in vitro study

Background Despite a revived interest in fat grafting procedures, clinicians still fail to demonstrate clearly the in vivo behavior of fat grafts as a dynamic tissue substitute. However, the basic principles in cellular biology teach us that cells can survive and develop, provided that a structural matrix exists that directs their behavior. The purpose of this in vitro study was to analyze that behavior of crude fat grafts, cultured on a three-dimensional laminin-rich matrix. Methods Nonprocessed, human fat biopsy specimens (approximately 1 mm) were inoculated on Matrigel-coated wells to which culture medium was added. The control group consisted of fat biopsy specimens embedded in medium alone. The cellular proliferation pattern was followed over 6 weeks. Additional cultures of primary generated cellular spheroids were performed and eventually subjected to adipogenic differentiation media. Results A progressive outgrowth of fibroblast-like cells from the core fat biopsy specimen was observed in both groups. Within the Matrigel group, an interconnecting three-dimensional network of spindle-shaped cells was established. This new cell colony reproduced spheroids that functioned again as solitary sources of cellular proliferation. Addition of differentiation media resulted in lipid droplet deposition in the majority of generated cells, indicating the initial steps of adipogenic differentiation. Conclusions The authors noticed that crude, nonprocessed fat biopsy specimens do have considerable potential for future tissue engineering-based applications, provided that the basic principles of developmental, cellular biology are respected. Spontaneous in vitro expansion of the stromal cells present in fat grafts within autologous and injectable matrices could create "off-the-shelf" therapies for reconstructive procedures.

Soluble laminin and arginine-glycine-aspartic acid containing peptides differentially regulate type IV collagenase messenger RNA, activation, and localization in testicular cell culture

Rat testicular cells in culture produce several metalloproteinases including type IV collagenases (Sang et al. Biol Reprod 1990; 43:946-955, 956-964). We have now investigated the regulation of testicular cell type IV collagenase and other metalloprotemases in vitro. Soluble laminin stimulated Sertoli cell type IV collagenase mRNA levels. However, three peptides corresponding to different domains of the laminin molecule (CSRAKQAASIKVASADR, FALRGDNP, CLQDGDVRV) did not influence type IV collagenase mENA levels. Zyniographic analysis of medium collected from these cultures revealed that neither soluble laminin nor any of the peptides influenced 72-Wa type IV collagenase protein levels. However, peptide FALRGDNP resulted in both, a selective increase in two higher molecular-weight metalloprotemnases (83 kDa and 110 Wa and in an activation of the 72-Wa rat type IV collagenase. Interleukin-1, phorbol ester, testosterone, and FSH did not affect collagenase activation, lmmunocytochemical studies demonstrated that the addition of soluble laminin resulted in a redistribution of type IV collagenase from intracellular vesicles to the cell-substrate region beneath the cells. Peptide FALRGDNP induced a change from a vesicular to peripheral plasma membrane type of staining pattern. Zymography of plasma membrane preparations demonstrated triton-soluble gelatinases of 76 Wa, 83 Wa, and 110 Wa and a triton-insoluble gelatinase of 225 Wa, These results indicate that testicular cell type IV collagenase mRNA levels, enzyme activation, and distribution are influenced by laminin and RGD-containing peptides.

The construction of social identity in newly recruited nuclear engineering staff : a longitudinal study

This study examines the process by which newly recruited nuclear engineering and technical staff came to understand, define, think, feel and behave within a distinct group that has a direct contribution to the organization's overall emphasis on a culture of reliability and system safety. In the field of organizational behavior the interactive model of social identity formation has been recently proposed to explain the process by which the internalization of shared norms and values occurs, an element critical in identity formation. Using this rich model of organizational behavior we analyzed multiple sources of data from nine new hires over a period of three years. This was done from the time they were employed to investigate the construction of social identity by new entrants entering into a complex organizational setting reflected in the context of a nuclear facility. Informed by our data analyses, we found support for the interactive model of social identity development and report the unexpected finding that a newly appointed member's age and level of experience appears to influence the manner in which they adapt, and assimilate into their surroundings. This study represents an important contribution to the safety and reliability literature as it provides a rich insight into the way newly recruited employees enact the process by which their identities are formed and hence act, particularly under conditions of duress or significant organizational disruption in complex organizational settings.

An empirical study of the factors impacting on the involvement of clients in Saudi Arabian construction projects

This thesis is an empirical study of the factors impacting on the client involvement in Saudi Arabian construction government projects. The study investigated the impact of some factors that limited the client involvement and developed a framework called "client involvement interactive" to improve the client involvement practices in Saudi Arabian construction projects through the implementation of an implementable strategy elaborated.

Sertoli cells as paracrine modulators of DNA synthesis in rat peritubular myoid cells in culture

To determine whether Sertoli cells influence DNA synthesis by rat peritubular myoid cells in vitro, the effects of Sertoli cells on [3H]thymidine incorporation by peritubular myoid cells in a coculture situation were examined. Incubation of testicular peritubular myoid cells with Sertoli cells in coculture induced a significant increase in [3H]thymidine incorporation by peritubular myoid cells. This indicates a cell-cell cooperation between Sertoli and peritubular myoid cells in the testis in terms of DNA synthesis. Secreted factors from Sertoli cells, as tested in a parabiotic culture situation, also increased [3H]thymidine incorporation by peritubular myoid cells. Moreover, in terms of total cellular protein, cocultures of Sertoli cells and peritubular myoid cells resulted in a significant increase when compared with the monocultures, and this coculture effect substituted for the stimulatory response of serum on peritubular myoid cell monoculture. This study investigated the cooperative role of Sertoli cells and peritubular myoid cells in paracrine regulation of testicular functions.

The alcohol culture of mature-aged women : similarities and differences across borders

Introduction: Relatively few attempts have been made to describe and understand women���s alcohol consumption beyond adolescence and young adulthood. In particular, there has been a lack of studies focusing on the alcohol culture that surrounds and guides mature-aged women���s drinking. As part of a larger cross-national comparison, the present study sought to address this gap by identifying the shared beliefs and values that impact on drinking outcomes among mature-aged women in Sweden and Australia. Method: The study was guided by an ethnographic methodology. To generate data, a series of semi-structured interview were conducted with 17 Australian (age = 45-57 years; M = 52.1, SD = 3.9) and 19 Swedish (age = 45-58 years; M = 52.2, SD = 4.8) women. All interviews were transcribed verbatim and thematically analysed. Results: With age, the focus of alcohol as a single purpose vehicle for intoxication had given way to a focus on the enjoyment and ritual of drinking itself; taste had become increasingly important and alcohol was strongly associated with pleasurable environments and experiences. The view of alcohol as a taste experience was particularly pronounced among the Swedish women, with alcohol (most commonly wine) often seen as inseparable from food. Among the Swedish women, this view of alcohol was also associated with a strong de-emphasis of the pharmacological effects of alcohol. In contrast, several Australian women understood and used alcohol as relief for anxiety and stress. Moderate drinking was linked to the social construction of both age and gender in the two samples, while heavy or abusive alcohol consumption was associated with strong proscriptive norms and stigma. Conclusions: Australian and Swedish women share a number of beliefs and values around alcohol, however, these findings also show unique country-level differences. Implications for drinking outcomes are discussed.

Sense of home, sense of place : a morpho-typological comparative study of Florida and Queensland

Cities in the 21st century have become layered and complex systems not only in terms of physical form, but also social and cultural structure. Consolidated tools to analyze the urban environment have today to be improved including a strong interdisciplinary perspective in order to understand and manage the unprecedented complexity our cities are facing. Redevelopments, new estates, internal and external migrations are all dynamics which are deeply modifying the built environment directly or indirectly also affecting local identity, culture and social structure. This paper investigates the relationship between urban form and social behaviors, with particular attention to the perception of the built environment and its use by long term residents, recent migrants as well as tourists. A comparative study is suggested between South East Queensland and Florida; this two regions share common features such as subtropical climate, similar lifestyle, leisure cities and canal estates. Neighborhoods on the Gold and Sunshine Coasts have been designed using the communities of Florida, such as Celebration or Seaside, as models. These regions share also significant migration processes, similar social problems and high crime rates, which directly affect the local economies. Comparing Florida and SEQ could provide an understanding of different strategies adopted and how urban development and lifestyle can be managed maintaining social equity and security. This study, investigates people���s perception of built form and how this affects the use of public space. The relationship between built environment and social behaviour has been previously investigated, for example by environmental psychology; the innovation proposed by this research is to study the perception of place in leisure cities at multiple levels. Locals, migrants and tourists have different understanding of the built form in the same location; this understanding affects the use of space and the attitude to visit or avoid some precincts. The research methodology integrates traditional morpho-typological investigations with qualitative methods; data are collected in the first phase through online surveys about perception of urban forms. Findings guide then the selection of neighbourhoods to be investigated in detail through questionnaires and Nolli maps, specifying morphological regions as well as recurrent building typologies. A final phase includes interviews with selected stakeholders. Major urban projects are discussed addressing how they are used and perceived by locals, migrants or tourists; the comparison between SEQ and Florida allows the identification of strategies to address migration issues in both regions with particular attention to urban form and placemaking dynamics.

Straminipilan protists : a study of their taxonomic position, morphology and abiotic stress-mediated gene expression

Thraustochytrids have become of considerable industrial and scientific interest in the past decade due to their health benefits. They have been proven to be the principle source in marine and estuarine fish diets with high percentage of long chain (LC) or polyunsaturated fatty acids (PUFA). Therefore, the oil extracted from fish for human document.forms[0].elements[13].select();consumption is rich in PUFA with high omega-3 fatty acid content. Docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) of all of the omega-3 fatty acids, are considered beneficial essential oils for humans with a wide range of health benefits. These include brain and neural development in infants, general wellbeing of adults and drug delivery through precursor molecules. They have become one of the most extensively studied organisms for industrial oil preparations as PUFA extraction from fish becomes less profitable. Many forms of these Thraustochytrid oils are being trialled for human consumption all over the world. In Australia, there has been little research performed on these organisms in the past ten years. A few Australian studies have been conducted in the form of comparative studies related to PUFA production within the related genera, but not focussed on their identification or cellular and genomic characterisation. Therefore, the main aim of this study was to investigate the morphological and genetic characteristics of Australian Thraustochytrids in order to aid in their identification and characterisation, as well as to better understand the effect of environmental conditions in the regulation of PUFA production. It was also noted that there was a knowledge gap in the preservation and total genomic DNA extraction of these organisms for the purposes of scientific research. The cryopreservation of these organisms for studies around the world follows existing generic methods. However, it is well understood that many of these generic methods attract not only high costs for chemicals, but also uses considerable storage space and other resources, all of which can be improved with new or modified approaches. In this context, a simple and inexpensive bead preservation method is described, without compromising the storage shelf life. We also describe, for the first time, the effects of culture age on the successful cryopreservation of Thraustochytrids. It was evident in the literature that DNA and RNA extractions for molecular and genetic studies of Thraustochytrids follow the classical phenol-chloroform extraction methods. It was also observed that modern protocols failed to avoid the use of phenol-chloroform rather than improving preparation and cell disruption. In order to provide a high quantity and quality DNA extraction, a modified protocol has been introduced that employs the use of modern commercial extraction kits and standard laboratory equipment. Thraustochytrids have been shown to be highly conserved in their 18S rDNA gene sequences, which is used as the current standard for identification. It was demonstrated that the 18S rDNA gene sequence limits the recognition of closely related genera or within the genera from each member. Therefore, it was proposed that another profile, such as a randomly amplified polymorphic DNA (RAPD) based profiling system, be tested for use in the characterisation of Thraustochytrids. The RAPD profiles were shown to provide a unique DNA fingerprint for each isolate and small variations in their genome were able to be detected. This method involved the use of a minimum number of standard arbitrary primers and with an increase in the number of different primers used, a very high discrimination between organisms could be achieved. However, the method was not suitable for taxonomic purposes because the results did not correlate with other taxonomic features such as morphology. Another knowledge gap was found with respect to Australian Thraustochytrid growth characteristics, in that these had not been recorded and published. In order to rectify this, a record of colony and microscopic features of 12 selected isolates was performed. The results of preliminary studies indicated that further microbiological and biochemical studies are needed for full characterisation of these organisms. This information is of great importance to bio-prospecting of new Thraustochytrids from Australian ecosystems and would allow for their accurate identification, and so permit the prediction of their PUFA capability by comparison with related genera/species. It was well recognized that environmental stress plays a role in the PUFA production and is mainly due to the reactive oxygen species as abiotic stress (Chiou et al., 2001; Okuyama et al., 2008; Shabala et al., 2009; Shabala et al., 2001). In this aspect, this study makes the first attempt towards better understanding of this phenomenon by way of the use of real-time PCR for the detection of environmental effects on the regulation of PUFA production. Three main environmental conditions including temperature, pH and oxygen availability were monitored as stress inducers. In summary, this study provides novel approaches for the preservation and handling of Thraustochytrids, their molecular biological features, taxonomy, characterisation and responses to environmental factors with respect to their oil production enzymes. The information produced from this study will prove to be vital for both industrial and scientific investigations in the future.

Practices of literary tourism : an Australian case study

Purpose To present the results of tests for the development of literary trails for domestic visitors and tourists in Brisbane, Queensland, and to situate these findings in the context of recent state government policy changes in relation to culture, community engagement and the environment. Design Broadly cultural studies: the article analyses changes in international and national cultural tourism and Queensland based issues before presenting the research findings. Findings a gap in tourist and cultural development models exists for the implementation of a network of sustainable literary trails in Brisbane--this model can be extended to regions around the state to meet the demands of the new tourist. Limitations Queensland weather and Australian distance which will require a regional approach that networks with transport and community hubs. Practical implications the research has produced new software for the use of self-guided walks; the locations for two specific area trails; and the involvement of the State Library of Queensland as a ���hub��� for the trails. Substantial support exists for further development in advanced locative media and gaming. Social implications the research demonstrates the importance of developing a sense of place that relates to culture, literary history and community for tourists, as well as the potential for community engagement.

A comparative study of the proliferation and osteogenic differentiation of human periodontal ligament cells cultured on ��-TCP ceramics and demineralized bone matrix with or without osteogenic inducers in vitro

The repair of bone defects that result from periodontal diseases remains a clinical challenge for periodontal therapy. ��-tricalcium phosphate (��-TCP) ceramics are biodegradable inorganic bone substitutes with inorganic components that are similar to those of bone. Demineralized bone matrix (DBM) is an acid-extracted organic matrix derived from bone sources that consists of the collagen and matrix proteins of bone. A few studies have documented the effects of DBM on the proliferation and osteogenic differentiation of human periodontal ligament cells (hPDLCs). The aim of the present study was to investigate the effects of inorganic and organic elements of bone on the proliferation and osteogenic differentiation of hPDLCs using three-dimensional porous ��-TCP ceramics and DBM with or without osteogenic inducers. Primary hPDLCs were isolated from human periodontal ligaments. The proliferation of the hPDLCs on the scaffolds in the growth culture medium was examined using a Cell���Counting kit���8 (CCK-8) and scanning electron microscopy (SEM). Alkaline phosphatase (ALP) activity and the osteogenic differentiation of the hPDLCs cultured on the ��-TCP ceramics and DBM were examined in both the growth culture medium and osteogenic culture medium. Specific osteogenic differentiation markers were examined using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). SEM images revealed that the cells on the ��-TCP were spindle-shaped and much more spread out compared with the cells on the DBM surfaces. There were no significant differences observed in cell proliferation between the ��-TCP ceramics and the DBM scaffolds. Compared with the cells that were cultured on ��-TCP ceramics, the ALP activity, as well as the Runx2 and osteocalcin (OCN) mRNA levels in the hPDLCs cultured on DBM were significantly enhanced both in the growth culture medium and the osteogenic culture medium. The organic elements of bone may exhibit greater osteogenic differentiation effects on hPDLCs than the inorganic elements.

Risk management and high reliability : a case study of Australian airport management

This thesis investigated risk management and organisational reliability in airports from the perspective of complex sociotechnical systems (CSSs). Two research studies were undertaken, the first focusing on the processes by which disruptive events occur, are detected and responded to; the second exploring the presence of organisational reliability traits within airports. A key result of the studies was the development of new approach: the Critical Incident Disturbance Process model that detailed a means to understand and analyse how disruptions in complex CSSs might be influenced by vulnerability reduction and enhanced risk management. Further, this thesis identified and defined the concept of 'compartmentalised reliability' in complex sociotechnical systems, extending existing knowledge of high reliability theory.

Recognising emotional expressions of complaining customers : A cross-cultural study

Purpose This study aims to test service providers��� ability to recognise non-verbal emotions in complaining customers of same and different cultures. Design/methodology/approach In a laboratory study, using a between-subjects experimental design (n = 153), we tested the accuracy of service providers��� perceptions of the emotional expressions of anger, fear, shame and happiness of customers from varying cultural backgrounds. After viewing video vignettes of customers complaining (with the audio removed), participants (in the role of service providers) assessed the emotional state of the customers portrayed in the video. Findings Service providers in culturally mismatched dyads were prone to misreading anger, happiness and shame expressed by dissatisfied customers. Happiness was misread in the displayed emotions of both dyads. Anger was recognisable in the Anglo customers but not Confucian Asian, while Anglo service providers misread both shame and happiness in Confucian Asian customers. Research limitations/implications The study was conducted in the laboratory and was based solely on participant���s perceptions of actors��� non-verbal facial expressions in a single encounter. Practical implications Given the level of ethnic differences in developed nations, a culturally sensitive workplace is needed to foster effective functioning of service employee teams. Ability to understand cultural display rules and to recognise and interpret emotions is an important skill for people working in direct contact with customers. Originality/value This research addresses the lack of empirical evidence for the recognition of customer emotions by service providers and the impact of cross-cultural differences.

Better half-dead than read? The Mezzomorto cases and their implications for literary culture in the 1930s

This article focusses on the two libel cases arising from Brian Penton's review of Vivian Crockett's novel Mezzomorto for the Bulletin in 1934, viewing them as points of entry into Australian literary politics in the 1930s, and as windows on to one of the most enduring and interesting feuds in Australian literary culture, that between P.R. 'Inky' Stephensen, self-styled 'Bunyip Critic,' and Brian Penton, arch exponent of 'destructive criticism' and scourge of parochial pretension. The cases are particularly interesting for what they reveal about the evolving positions of two influential figures in Australian writing of the 1930s and 1940s. They also play in to contemporary debates about the state and status of 'literature' in Australia. And while Penton's biographer Patrick Buckridge avers that the cases did not impact on any of the larger contemporary literary issues (meaning censorship and free speech), a case may be made for the significance of the libel actions in the context of attempts to establish an industrial and cultural presence for a diverse range of Australian writing.

Challenges confronting career-changing beginning teachers : a qualitative study of professional scientists becoming science teachers

Recruitment of highly qualified science and mathematics graduates has become a widespread strategy to enhance the quality of education in the field of STEM. However, attrition rates are very high suggesting preservice education programs are not preparing them well for the career change. We analyse the experiences of professionals who are scientists and have decided to change careers to become teachers. The study followed a group of professionals who undertook a one-year preservice teacher education course and were employed by secondary schools on graduation. We examined these teachers��� experiences through the lens of self-determination theory, which posits autonomy, confidence and relatedness are important in achieving job satisfaction. The findings indicated that the successful teachers were able to achieve a sense of autonomy and confidence, and, in particular, had established strong relationships with colleagues. However, the unique challenges facing career-change professionals were often overlooked by administrators and colleagues. Opportunities to build a sense of relatedness in their new profession were often absent. The failure to establish supportive relationships was decisive in some teachers leaving the profession. The findings have implications for both pre-service and professional inservice programs and the role that administrators play in supporting career-change teachers.

Choosing an appropriate modelling framework for analysing multispecies co-culture cell biology experiments

In vitro cell biology assays play a crucial role in informing our understanding of the migratory, proliferative and invasive properties of many cell types in different biological contexts. While mono-culture assays involve the study of a population of cells composed of a single cell type, co-culture assays study a population of cells composed of multiple cell types (or subpopulations of cells). Such co-culture assays can provide more realistic insights into many biological processes including tissue repair, tissue regeneration and malignant spreading. Typically, system parameters, such as motility and proliferation rates, are estimated by calibrating a mathematical or computational model to the observed experimental data. However, parameter estimates can be highly sensitive to the choice of model and modelling framework. This observation motivates us to consider the fundamental question of how we can best choose a model to facilitate accurate parameter estimation for a particular assay. In this work we describe three mathematical models of mono-culture and co-culture assays that include different levels of spatial detail. We study various spatial summary statistics to explore if they can be used to distinguish between the suitability of each model over a range of parameter space. Our results for mono-culture experiments are promising, in that we suggest two spatial statistics that can be used to direct model choice. However, co-culture experiments are far more challenging: we show that these same spatial statistics which provide useful insight into mono-culture systems are insuffcient for co-culture systems. Therefore, we conclude that great care ought to be exercised when estimating the parameters of co-culture assays.