Long-term stability of adipose tissue generated from a vascularized pedicled fat flap inside a chamber

Background Numerous studies demonstrate the generation and short-term survival of adipose tissue; however, long-term persistence remains elusive. This study evaluates long-term survival and transferability of de novo adipose constructs based on a ligated vascular pedicle and tissue engineering chamber combination. Methods Defined adipose tissue flaps were implanted into rats in either intact or perforated domed chambers. In half of the groups, the chambers were removed after 10 weeks and the constructs transferred on their vascular pedicle to a new site, where they were observed for a further 10 weeks. In the remaining groups, the tissue construct was observed for 20 weeks inside the chamber. Tissue volume was assessed using magnetic resonance imaging and histologic measures, and constructs were assessed for stability and necrosis. Sections were assessed histologically and for proliferation using Ki-67. Results At 20 weeks, volume analysis revealed an increase in adipose volume from 0.04 ± 0.001 ml at the time of insertion into the chambers to 0.27 ± 0.004 ml in the closed and 0.44 ± 0.014 ml in the perforated chambers. There was an additional increase of approximately 10 to 15 percent in tissue volume in flaps that remained in chambers for 20 weeks, whereas the volume of the transferred tissue not in chambers remained unaltered. Histomorphometric assessment of the tissues documented no signs of hypertrophy, fat necrosis, or atypical changes of the newly generated tissue. Conclusion This study presents a promising new method of generating significant amounts of mature, vascularized, stable, and transferable adipose tissue for permanent autologous soft-tissue replacement.

MT1-MMP-dependent and -independent regulation of gelatinase A activation in long-term, ascorbate-treated fibroblast cultures: Regulation by fibrillar collagen

Human skin fibroblasts were cultured long-term in the presence of ascorbic acid to allow formation of a three-dimensional collagen matrix, and the effects of this on activation of secreted matrix metalloproteinase-2 (MMP-2) were examined. Accumulation of collagen over time correlated with increased levels of both mature MMP-2 and cell-associated membrane type 1-MMP (MT1-MMP), and subsequently increased mRNA levels for MT1-MMP, providing temporal resolution of the "nontranscriptional" and "transcriptional" effects of collagen on MT-1MMP functionality. MMP-2 activation by these cultures was blocked by inhibitors of prolyl-4-hydroxylase, or when fibroblasts derived from the collagen α1(I) gene-deficient Mov-13 mouse were used. MMP-2 activation by the Mov-13 fibroblasts was rescued by transfection of a full-length α1(I) collagen cDNA, and to our surprise, also by transfection with an α1(I) collagen cDNA carrying a mutation at the C-proteinase cleavage, which almost abrogated fibrillogenesis. Although studies with ascorbate-cultured MT1-MMP-/- fibroblasts showed that MT1-MMP played a significant role in the collagen-induced MMP-2 activation, a residual MT1-MMP-independent activation of MMP-2 was seen which resembled the level of MMP-2 activation persisting when wild-type fibroblasts were cultured in the presence of both ascorbic acid and MMP inhibitors. We were also unable to block this residual activation with inhibitors specific for serinyl, aspartyl, or cysteinyl enzymes.

Why are early maturing girls less active? Links between pubertal development, psychological well-being, and physical activity among girls at ages 11 and 13

Previous research has shown that early maturing girls at age I I have lower subsequent physical activity at age 13 in comparison to later maturing girls. Possible reasons for this association have not been assessed. This study examines girls' psychological response to puberty and their enjoyment of physical activity as intermediary factors linking pubertal maturation and physical activity. Participants included 178 girls who were assessed at age 11, of whom 168 were reassessed at age 13. All participants were non-Hispanic white and resided in the US. Three measures of pubertal development were obtained at age I I including Tanner breast stage, estradiol levels, and mothers' reports of girls' development on the Pubertal Development Scale (PDS). Measures of psychological well-being at ages I I and 13 included depression, global self-worth, perceived athletic competence, maturation fears, and body esteem. At age 13, girls' enjoyment of physical activity was assessed using the Physical Activity Enjoyment Scale and their daily minutes of moderate-to-vigorous physical activity (MVPA) were assessed using objective monitoring. Structural Equation Modeling was used to assess direct and indirect pathways between pubertal development at age I I and MVPA at age 13. In addition to a direct effect of pubertal development on MVPA, indirect effects were found for depression, global self-worth and maturity fears controlling for covariates. In each instance, more advanced pubertal development at age I I was associated with lower psychological wellbeing at age 13, which predicted lower enjoyment of physical activity at age 13 and in turn lower MVPA. Results from this study suggest that programs designed to increase physical activity among adolescent girls should address the self-consciousness and discontent that girls' experience with their bodies during puberty, particularly if they mature earlier than their peers, and identify activities or settings that make differences in body shape less conspicuous.

Host rather than graft origin of Matrigel-induced adipose tissue in the murine tissue-engineering chamber

We have recently shown that Matrigel-filled chambers containing fibroblast growth factor-2 (FGF2) and placed around an epigastric pedicle in the mouse were highly adipogenic. Contact of this construct with pre-existing tissue or a free adipose graft was required. To further investigate the mechanisms underpinning formation of new adipose tissue, we seeded these chambers with human adipose biopsies and human adipose-derived cell populations in severe combined immunodeficient mice and assessed the origin of the resultant adipose tissue after 6 weeks using species-specific probes. The tissues were negative for human-specific vimentin labeling, suggesting that the fat originates from the murine host rather than the human graft. This was supported by the strong presence of mouse-specific Cot-1 deoxyribonucleic acid labeling, and the absence of human Cot-1 labeling in the new fat. Even chambers seeded with FGF2/Matrigel containing cultured human stromal-vascular fraction (SVF) labeled strongly only for human vimentin in cells that did not have a mature adipocyte phenotype; the newly formed fat tissue was negative for human vimentin. These findings indicate that grafts placed in the chamber have an inductive function for neo-adipogenesis, rather than supplying adipocyte-precursor cells to generate the new fat tissue, and preliminary observations implicate the SVF in producing inductive factors. This surprising finding opens the door for refinement of current adipose tissue-engineering approaches.

Noisy Bluetooth traffic data?

Traffic state estimation in an urban road network remains a challenge for traffic models and the question of how such a network performs remains a difficult one to answer for traffic operators. Lack of detailed traffic information has long restricted research in this area. The introduction of Bluetooth into the automotive world presented an alternative that has now developed to a stage where large-scale test-beds are becoming available, for traffic monitoring and model validation purposes. But how much confidence should we have in such data? This paper aims to give an overview of the usage of Bluetooth, primarily for the city-scale management of urban transport networks, and to encourage researchers and practitioners to take a more cautious look at what is currently understood as a mature technology for monitoring travellers in urban environments. We argue that the full value of this technology is yet to be realised, for the analytical accuracies peculiar to the data have still to be adequately resolved.

Association of MMP-2 activation potential with metastatic progression in human breast cancer cell lines independent of MMP-2 production

Background: Expression of matrix metalloproteinase-2 (MMP-2), the 72-kd type IV collagenase/gelatinase, by cancer cells has been implicated in metastasis through cancer cell invasion of basement membranes mediated by degradation of collagen IV. However, the abundance of this latent proenzyme in normal tissues and fluids suggests that MMP-2 proenzyme utilization is limited by its physiological activation rather than expression alone. We previously reported activation of this proenzyme by normal and malignant fibroblastoid cells cultured on collagen I (vitrogen) gels. Purpose: Our purposes in this study were 1) to determine whether MMP-2 activation is restricted to the more invasive human breast cancer cell lines and 2) to localize the activating mechanism. Methods: Zymography was used to monitor MMP-2 activation through detection of latent MMP-2 (72 kd) and mature species of smaller molecular weight (59 or 62 kd). Human breast cancer cell lines cultured on plastic, vitrogen, and other matrices were thus screened for MMP- 2 activation. Collagen I-cultured cells were exposed to cycloheximide, a protein synthesis inhibitor, or to protease inhibitors to determine the nature of the MMP-2-activating mechanism. Triton X-114 (TX-114) detergent extracts from cells cultured on collagen I or plastic were incubated with latent MMP-2 and analyzed by zymography to localize the MMP-2 activator. Results: MMP-2 activation was only induced by collagen I culture in the more aggressive, highly invasive estrogen receptor-negative, vimentin-positive human breast cancer cell lines (Hs578T, MDA-MB-436, BT549, MDA-MB-231, MDA- MB-435, MCF-7(ADR)) and was independent of MMP-2 production. MMP-2 activation was detected in cells cultured on collagen I gels but not in those cultured on gelatin gels, Matrigel, or thin layers of collagen I or IV, gelatin, or fibronectin. Collagen-induced activation was specific for the enzyme species MMP-2, since MMP-9, the 92-kd type IV collagenase/gelatinase, was not activatable under similar conditions. MMP-2 activation was inhibited by cycloheximide and was sensitive to a metalloproteinase inhibitor but not to aspartyl, serine, or cysteinyl protease inhibitors. MMP-2 activation was detected in the hydrophobic, plasma membrane-enriched, TX-114 extracts from invasive collagen I-cultured cells. Conclusion: Collagen I-induced MMP-2 activation is restricted to highly invasive estrogen receptor-negative, vimentin-positive human breast cancer cell lines, is independent of MMP-2 production, and is associated with metastatic potential. Our findings are consistent with plasma membrane localization of the activator. Implications: The MMP-2 activation mechanism may represent a new target for diagnosis, prognosis, and treatment of human breast cancer.

Management of project knowledge at various maturity levels in PMO, a theoretical framework

Management of project knowledge is a critical factor for project success. Project Management Office (PMO) is a unit within organisations to centrally facilitate, manage and control organisational project for improving the rate of project success. Due to increasing interest of developing PMO, the Project Management Maturity Model (PMMM) has been proposed to develop PMOs gradually. The PMMM contributes to evolvement of PMO from immature to mature level through addressing appropriate PM practices. Despite the importance of project knowledge, it has not been extensively investigated in project environments. In addition, the existing PMMMs not only do not address management of project knowledge, but also they recommend little criteria to assess the maturity of PMO from KM point of view. The absence of KM discussion in current PMMMs was defined as the subject of a research project in order for addressing KM practices at various maturity levels of PMO. In order to address the mentioned gap, a framework has been developed based on the current discussions of both PM and KM. The proposed framework comprises three premises: KM processes and practices, PMMM, and KM Maturity Model (KMMM). The incorporation of KMMM practices at various maturity levels of PMO is one of the significance of this framework. It proposes numbers of KM strategies, processes, and practices to address project knowledge management at various levels PMO. This framework shall be useful guidance for developing PMOs from KM perspective. In other words, it contributes to management of project knowledge, as a key for project success. The proposed framework follows the process-based approach and it could be employed alongside the current PMMMs for PMO development. This paper presents the developed framework, theoretical background, premises, proposed KM practices, and processes to be employed in Project-based Organisations and PMOs. This framework has been examined at numbers of case studies with different maturity levels. The case studies outcomes, which will be subjects for future papers, have not shown any significant contradiction yet, however, more investigations are being conducted to validate the proposed framework.

Management of project knowledge in a project-based organisation : a case study of research enterprise

Knowledge Management (KM) is vital factor to successfully undertake projects. The temporary nature of projects necessitates employing useful KM practices to reduce any issues such as knowledge leakiness and rework. The Project Management Office (PMO) is a unit within organisations to facilitate and oversee organisational projects. Project Management Maturity Models (PMMM) show the development of PMOs from immature to mature levels. The existing PMMMs have focused on discussing Project Management (PM) practices, however, the management of project knowledge is yet to be addressed, at various levels of maturity. A research project was undertaken to investigate the mentioned gap for addressing KM practices at the existing PMMMs. Due to the exploratory and inductive nature of this research, qualitative methods using case studies were chosen as the research methodology to investigate the problem in the real world. In total, three cases selected from different industries: research; mining and government organisations, to provide broad categories for research and research questions were examined using the developed framework. This paper presents the findings from the investigation of the research organisation with the lowest level of maturity. From KM process point of view, knowledge creation and capturing are the most important processes, while knowledge transferring and reusing received less attention. In addition, it was revealed that provision of “knowledge about client” and “project management knowledge” are the most important types of knowledge that are required at this level of maturity. The results also revealed that PMOs with higher maturity level have better knowledge management, however, some improvement is needed. In addition, the importance of KM processes varies at different levels of maturity. In conclusion, the outcomes of this paper could provide powerful guidance to PMOs at lowest level of maturity from KM point of view.

Bike share's impact on car use : evidence from the United States, Great Britain, and Australia

There are currently more than 700 cities operating bike share programs. Purported benefits of bike share include flexible mobility, physical activity, reduced congestion, emissions and fuel use. Implicit or explicit in the calculation of program benefits are assumptions regarding the modes of travel replaced by bike share journeys. This paper examines the degree to which car trips are replaced by bike share, through an examination of survey and trip data from bike share programs in Melbourne, Brisbane, Washington, D.C., London, and Minneapolis/St. Paul. A secondary and unique component of this analysis examines motor vehicle support services required for bike share fleet rebalancing and maintenance. These two components are then combined to estimate bike share’s overall contribution to changes in vehicle kilometers traveled. The results indicate an estimated reduction in motor vehicle use due to bike share of approx. 90,000 km per annum in Melbourne and Minneapolis/St. Paul and 243,291 km for Washington, D.C. London’s bike share program however recorded an additional 766,341 km in motor vehicle use. This was largely due to a low car mode substitution rate and substantial truck use for rebalancing of bicycles. As bike share programs mature, evaluation of their effectiveness in reducing car use may become increasingly important. Researchers can adapt the analytical approach proposed in this paper to assist in the evaluation of current and future bike share programs.

Funding emergency care : Australian style

The ongoing challenge for ED leaders is to remain abreast of system-wide changes that impact on the day-to-day management of their departments. Changes to the funding model creates another layer of complexity and this introductory paper serves as the beginning of a discussion about the way in which EDs are funded and how this can and will impact on business decisions, models of care and resource allocation within Australian EDs. Furthermore it is evident that any funding model today will mature and change with time, and moves are afoot to refine and contextualise ED funding over the medium term. This perspective seeks to provide a basis of understanding for our current and future funding arrangements in Australian EDs.

Shared generation of pseudo-random functions

In Crypto’95, Micali and Sidney proposed a method for shared generation of a pseudo-random function f(·) among n players in such a way that for all the inputs x, any u players can compute f(x) while t or fewer players fail to do so, where 0⩽tseeds S={s1,…,sd} of a poly-random collection of functions, among the n players, each player gets a subset of S, in such a way that any u players together hold all the secret seeds in S while any t or fewer players will lack at least one element from S. The pseudo-random function is then computed as where fsi(·)'s are poly-random functions. One question raised by Micali and Sidney is how to distribute the secret seeds satisfying the above condition such that the number of seeds, d, is as small as possible. In this paper, we continue the work of Micali and Sidney. We first provide a general framework for shared generation of pseudo-random function using cumulative maps. We demonstrate that the Micali–Sidney scheme is a special case of this general construction. We then derive an upper and a lower bound for d. Finally we give a simple, yet efficient, approximation greedy algorithm for generating the secret seeds S in which d is close to the optimum by a factor of at most u ln 2.

Shared generation of pseudo-random functions with cumulative maps

In Crypto’95, Micali and Sidney proposed a method for shared generation of a pseudo-random function f(·) among n players in such a way that for all the inputs x, any u players can compute f(x) while t or fewer players fail to do so, where 0 ≤ t < u ≤ n. The idea behind the Micali-Sidney scheme is to generate and distribute secret seeds S = s1, . . . , sd of a poly-random collection of functions, among the n players, each player gets a subset of S, in such a way that any u players together hold all the secret seeds in S while any t or fewer players will lack at least one element from S. The pseudo-random function is then computed as where f s i (·)’s are poly-random functions. One question raised by Micali and Sidney is how to distribute the secret seeds satisfying the above condition such that the number of seeds, d, is as small as possible. In this paper, we continue the work of Micali and Sidney. We first provide a general framework for shared generation of pseudo-random function using cumulative maps. We demonstrate that the Micali-Sidney scheme is a special case of this general construction.We then derive an upper and a lower bound for d. Finally we give a simple, yet efficient, approximation greedy algorithm for generating the secret seeds S in which d is close to the optimum by a factor of at most u ln 2.

Harvesting as an alternative to burning for managing Spinifex Grasslands in Australia

Sustainable harvesting of grasslands can buffer large scale wildfires and the harvested biomass can be used for various products. Spinifex (Triodia spp.) grasslands cover ≈30% of the Australian continent and form the dominant vegetation in the driest regions. Harvesting near settlements is being considered as a means to reduce the occurrence and intensity of wildfires and to source biomaterials for sustainable desert living. However, it is unknown if harvesting spinifex grasslands can be done sustainably without loss of biodiversity and ecosystem function. We examined the trajectory of plant regeneration of burned and harvested spinifex grassland, floristic diversity, nutrient concentrations in soil and plants, and seed germination in controlled ex situ conditions. After two to three years of burning or harvesting in dry or wet seasons, species richness, diversity, and concentrations of most nutrients in soil and leaves of regenerating spinifex plants were overall similar in burned and harvested plots. Germination tests showed that 20% of species require fire-related cues to trigger germination, indicating that fire is essential for the regeneration of some species. Further experimentation should evaluate these findings and explore if harvesting and intervention, such as sowing of fire-cued seeds, allow sustainable, localised harvesting of spinifex grasslands.

PTRF/cavin-1 neutralizes non-caveolar caveolin-1 microdomains in prostate cancer

Caveolin-1 has a complex role in prostate cancer and has been suggested to be a potential biomarker and therapeutic target. As mature caveolin-1 resides in caveolae, invaginated lipid raft domains at the plasma membrane, caveolae have been suggested as a tumor-promoting signaling platform in prostate cancer. However, caveola formation requires both caveolin-1 and cavin-1 (also known as PTRF; polymerase I and transcript release factor). Here, we examined the expression of cavin-1 in prostate epithelia and stroma using tissue microarray including normal, non-malignant and malignant prostate tissues. We found that caveolin-1 was induced without the presence of cavin-1 in advanced prostate carcinoma, an expression pattern mirrored in the PC-3 cell line. In contrast, normal prostate epithelia expressed neither caveolin-1 nor cavin-1, while prostate stroma highly expressed both caveolin-1 and cavin-1. Utilizing PC-3 cells as a suitable model for caveolin-1-positive advanced prostate cancer, we found that cavin-1 expression in PC-3 cells inhibits anchorage-independent growth, and reduces in vivo tumor growth and metastasis in an orthotopic prostate cancer xenograft mouse model. The expression of α-smooth muscle actin in stroma along with interleukin-6 (IL-6) in cancer cells was also decreased in tumors of mice bearing PC-3-cavin-1 tumor cells. To determine whether cavin-1 acts by neutralizing caveolin-1, we expressed cavin-1 in caveolin-1-negative prostate cancer LNCaP and 22Rv1 cells. Caveolin-1 but not cavin-1 expression increased anchorage-independent growth in LNCaP and 22Rv1 cells. Cavin-1 co-expression reversed caveolin-1 effects in caveolin-1-positive LNCaP cells. Taken together, these results suggest that caveolin-1 in advanced prostate cancer is present outside of caveolae, because of the lack of cavin-1 expression. Cavin-1 expression attenuates the effects of non-caveolar caveolin-1 microdomains partly via reduced IL-6 microenvironmental function. With circulating caveolin-1 as a potential biomarker for advanced prostate cancer, identification of the molecular pathways affected by cavin-1 could provide novel therapeutic targets.

Ethnic cluster lifecycle : a case of Chinese restaurants in South East Queensland

Immigrant entrepreneurs tend to start businesses within their ethnic enclave (EE), as it is an integral part of their social and cultural context and the location where ethnic resources reside (Logan, Alba, & Stults, 2003). Ethnic enclaves can be seen as a form of geographic cluster, China Towns are exemplar EEs, easily identified by the clustering of Chinese restaurants and other ethnic businesses in one central location. Studies on EE thus far have neglected the life cycles stages of EE and its impact on the business experiences of the entrepreneurs. In this paper, we track the formation, growth and decline of an EE. We argue that EE is a special industrial cluster and as such it follows the growth conditions proposed by the cluster life cycle theory (Menzel & Fornahl, 2009). We report a mixed method study of Chinese Restaurants in South East Queensland. Based on multiple sources of data, we concluded that changes in government policies leading to a sharp increase of immigrant numbers from a distinctive culture group can lead to the initiation and growth of the EE. Continuous incoming of new immigrants and increase competition within the cluster mark the mature stage of the EE, making the growth condition more favourable “inside” the cluster. A decline in new immigrants from the same ethnic group and the increased competition within the EE may eventually lead to the decline of such an industrial cluster, thus providing more favorable condition for growth of business outside the cluster.