Mechanical stimulation of the pro-angiogenic capacity of human fracture haematoma: Involvement of VEGF mechano-regulation

Compromised angiogenesis appears to be a major limitation in various suboptimal bone healing situations. Appropriate mechanical stimuli support blood vessel formation in vivo and improve healing outcomes. However, the mechanisms responsible for this association are unclear. To address this question, the paracrine angiogenic potential of early human fracture haematoma and its responsiveness to mechanical loading, as well as angiogenic growth factors involved, were investigated in vitro. Human haematomas were collected from healthy patients undergoing surgery within 72. h after bone fracture. The haematomas were embedded in a fibrin matrix, and cultured in a bioreactor resembling the in vivo conditions of the early phase of bone healing (20 compression, 1. Hz) over 3. days. Conditioned medium (CM) from the bioreactor was then analyzed. The matrices were also incubated in fresh medium for a further 24. h to evaluate the persistence of the effects. Growth factor (GF) concentrations were measured in the CM by ELISAs. In vitro tube formation assays were conducted on Matrigel with the HMEC-1 cell line, with or without inhibition of vascular endothelial growth factor receptor 2 (VEGFR2). Cell numbers were quantified using an MTS test. In vitro endothelial tube formation was enhanced by CM from haematomas, compared to fibrin controls. The angiogenesis regulators, vascular endothelial growth factor (VEGF) and transforming growth factor β1 (TGF-β1), were released into the haematoma CM, but not angiopoietins 1 or 2 (Ang1, 2), basic fibroblast growth factor (bFGF) or platelet-derived growth factor (PDGF). Mechanical stimulation of haematomas, but not fibrin controls, further increased the induction of tube formation by their CM. The mechanically stimulated haematoma matrices retained their elevated pro-angiogenic capacity for 24. h. The pro-angiogenic effect was cancelled by inhibition of VEGFR2 signalling. VEGF concentrations in CM tended to be elevated by mechanical stimulation; this was significant in haematomas from younger, but not from older patients. Other GFs were not mechanically regulated. In conclusion, the paracrine pro-angiogenic capacity of early human haematomas is enhanced by mechanical stimulation. This effect lasts even after removing the mechanical stimulus and appears to be VEGFR2-dependent.

A multilevel investigation into the effects of the philosophical community of inquiry on 6th grade students' reading comprehension, interest in maths, self-esteem, pro-social behaviours and emotional well-being

This investigation measured the effects of a one year participation in an Australian Philosophical Community of Inquiry program on 280 sixth grade students' reading comprehension, interest in maths, self-esteem, social behaviours, and emotional well-being. A multilevel model for change was used to detect differences in the response variables, between a quasi-experimental group and comparison group. Results showed that, for participants, reading comprehension significantly increased while interest in maths decreased. No differences between the groups were found for pro-social behaviour and emotional well-being. Self esteem, however, declined for participants while nonparticipants' self esteem increased.

Selection et controle de modes de deplacement pour un robot mobile autonome en environnements naturels

Autonomous navigation and locomotion of a mobile robot in natural environments remain a rather open issue. Several functionalities are required to complete the usual perception/decision/action cycle. They can be divided in two main categories : navigation (perception and decision about the movement) and locomotion (movement execution). In order to be able to face the large range of possible situations in natural environments, it is essential to make use of various kinds of complementary functionalities, defining various navigation and locomotion modes. Indeed, a number of navigation and locomotion approaches have been proposed in the literature for the last years, but none can pretend being able to achieve autonomous navigation and locomotion in every situation. Thus, it seems relevant to endow an outdoor mobile robot with several complementary navigation and locomotion modes. Accordingly, the robot must also have means to select the most appropriate mode to apply. This thesis proposes the development of such a navigation/locomotion mode selection system, based on two types of data: an observation of the context to determine in what kind of situation the robot has to achieve its movement and an evaluation of the behavior of the current mode, made by monitors which influence the transitions towards other modes when the behavior of the current one is considered as non satisfying. Hence, this document introduces a probabilistic framework for the estimation of the mode to be applied, some navigation and locomotion modes used, a qualitative terrain representation method (based on the evaluation of a difficulty computed from the placement of the robot's structure on a digital elevation map), and monitors that check the behavior of the modes used (evaluation of rolling locomotion efficiency, robot's attitude and configuration watching. . .). Some experimental results obtained with those elements integrated on board two different outdoor robots are presented and discussed.

Preliminary characterisation of tumor necrosis factor alpha and interleukin-10 responses to chlamydia pecorum infection in the koala (Phascolarctos cinereus)

Debilitating infectious diseases caused by Chlamydia are major contributors to the decline of Australia's iconic native marsupial species, the koala (Phascolarctos cinereus). An understanding of koala chlamydial disease pathogenesis and the development of effective strategies to control infections continue to be hindered by an almost complete lack of species-specific immunological reagents. The cell-mediated immune response has been shown to play an influential role in the response to chlamydial infection in other hosts. The objective of this study, hence, was to provide preliminary data on the role of two key cytokines, pro-inflammatory tumour necrosis factor alpha (TNFα) and anti-inflammatory interleukin 10 (IL10), in the koala Chlamydia pecorum response. Utilising sequence homology between the cytokine sequences obtained from several recently sequenced marsupial genomes, this report describes the first mRNA sequences of any koala cytokine and the development of koala specific TNFα and IL10 real-time PCR assays to measure the expression of these genes from koala samples. In preliminary studies comparing wild koalas with overt chlamydial disease, previous evidence of C. pecorum infection or no signs of C. pecorum infection, we revealed strong but variable expression of TNFα and IL10 in wild koalas with current signs of chlamydiosis. The description of these assays and the preliminary data on the cell-mediated immune response of koalas to chlamydial infection paves the way for future studies characterising the koala immune response to a range of its pathogens while providing reagents to assist with measuring the efficacy of ongoing attempts to develop a koala chlamydial vaccine.

Immunization with a MOMP-based vaccine protects mice against a Pulmonary Chlamydia challenge and identifies a disconnection between infection and pathology

Chlamydia pneumoniae is responsible for up to 20% of community acquired pneumonia and can exacerbate chronic inflammatory diseases. As the majority of infections are either mild or asymptomatic, a vaccine is recognized to have the greatest potential to reduce infection and disease prevalence. Using the C. muridarum mouse model of infection, we immunized animals via the intranasal (IN), sublingual (SL) or transcutaneous (TC) routes, with recombinant chlamydial major outer membrane protein (MOMP) combined with adjuvants CTA1-DD or a combination of cholera toxin/CpG-oligodeoxynucleotide (CT/CpG). Vaccinated animals were challenged IN with C. muridarum and protection against infection and pathology was assessed. SL and TC immunization with MOMP and CT/CpG was the most protective, significantly reducing chlamydial burden in the lungs and preventing weight loss, which was similar to the protection induced by a previous live infection. Unlike a previous infection however, these vaccinations also provided almost complete protection against fibrotic scarring in the lungs. Protection against infection was associated with antigen-specific production of IFNγ, TNFα and IL-17 by splenocytes, however, protection against both infection and pathology required the induction of a similar pro-inflammatory response in the respiratory tract draining lymph nodes. Interestingly, we also identified two contrasting vaccinations capable of preventing infection or pathology individually. Animals IN immunized with MOMP and either adjuvant were protected from infection, but not the pathology. Conversely, animals TC immunized with MOMP and CTA1-DD were protected from pathology, even though the chlamydial burden in this group was equivalent to the unimmunized controls. This suggests that the development of pathology following an IN infection of vaccinated animals was independent of bacterial load and may have been driven instead by the adaptive immune response generated following immunization. This identifies a disconnection between the control of infection and the development of pathology, which may influence the design of future vaccines.

Panem et circenses : law, law and power in The Hunger Games

Like many cautionary tales, The Hunger Games takes as its major premise an observation about contemporary society, measuring its ballistic arc in order to present graphically its logical conclusions. The Hunger Games gazes back to the panem et circenses of Ancient Rome, staring equally cynically forward, following the trajectory of reality television to its unbearably barbaric end point – a sadistic voyeurism for an effete elite of consumers. At each end of the historical spectrum (and in the present), the prevailing social form is Arendt’s animal laborans. Consumer or consumed, Panem’s population is (with the exception of the inner circle) either deprived of the possibility of, or distracted from, political action. Within the confines of the Games themselves, Law is abandoned or de‐realised: Law – an elided Other in the pseudo‐Hobbesian nightmare that is the Arena. The Games are played out, as were gladiatorial combats and other diversions of the Roman Empire, against a background resonant of Juvenal’s concern for his contemporaries’ attachment to short term gratification at the expense the civic virtues of justice and caring which are (or would be) constitutive of a contemporary form of Arendt’s homo politicus. While the Games are, on their face, ‘reality’ they are (like the realities presented in contemporary reality television) a simulated reality, de‐realised in a Foucauldian set design constructed as a distraction for Capitol, and for the residents of the Districts, a constant reminder of their subservience to Capitol. Yet contemporary Western culture, for which manipulative reality TV is but a symptom of an underlying malaise, is inscribed at least as an incipient Panem, Its public/political space is diminished by the effective slavery of the poor, the pre‐occupation with and distractions of materiality and modern media, and the increasing concentration of power/wealth into a smaller proportion of the population.

Consultant engagement for computer system selection: a pro-active client role in small enterprises

Because of their limited number of senior positions and fewer alternative career paths, small businesses have a more difficult time attracting and retaining skilled information systems (IS) staff and are thus dependent upon external expertise. Small businesses are particularly dependent on outside expertise when first computerizing. Because small businesses suffer from severe financial constraints. it is often difficult to justify the cost of custom software. Hence. for many small businesses, engaging a consultant to help with identifying suitable packaged software and related hardware, is their first critical step toward computerization. This study explores the importance of proactive client involvement when engaging a consultant to assist with computer system selection in small businesses. Client involvement throughout consultant engagement is found to be integral to project success and frequently lacking due to misconceptions of small businesses regarding their role. Small businesses often overestimate the impact of consultant and vendor support in achieving successful computer system selection and implementation. For consultant engagement to be successful, the process must be viewed as being directed toward the achievement of specific organizational results where the client accepts responsibility for direction of the process.

Toll-like receptor and pro-inflammatory cytokine expression during prolonged hyperinsulinaemia in horses : implications for laminitis

Equine laminitis, a disease of the lamellar structure of the horse’s hoof, can be incited by numerous factors that include inflammatory and metabolic aetiologies. However, the role of inflammation in hyperinsulinaemic laminitis has not been adequately defined. Tolllike receptor (TLR) activation results in up-regulation of inflammatory pathways and the release of pro-inflammatory cytokines, including interleukin-6 (IL-6) and tumour necrosis factor-alpha (TNF-�), and may be a pathogenic factor in laminitis. The aim of this study was to determine whether TLR4 expression and subsequent pro-inflammatory cytokine production is increased in lamellae and skeletal muscle during equine hyperinsulinaemia. Standardbred horses were treated with either a prolonged, euglycaemic hyperinsulinaemic clamp (p-EHC) or a prolonged, glucose infusion (p-GI), which induced marked and moderate hyperinsulinaemia, respectively. Age-matched control horses were treated simultaneously with a balanced electrolyte solution. Treated horses developed clinical (p-EHC) or subclinical (p-GI) laminitis, whereas controls did not. Skeletal muscle and lamellar protein extracts were analysed by Western blotting for TLR4, IL-6, TNF-� and suppressor of cytokine signalling 3 (SOCS3) expression. Lamellar protein expression of TLR4 and TNF-�, but not IL-6, was increased by the p-EHC, compared to control horses. A significant positive correlation was found between lamellar TLR4 and SOCS3. Skeletal muscle protein expression of TLR4 signalling parameters did not differ between control and p-EHC-treated horses. Similarly, the p-GI did not result in up-regulation of lamellar protein expression of any parameter. The results suggest that insulin-sensitive tissues may not accurately reflect lamellar pathology during hyperinsulinaemia. While TLR4 is present in the lamellae, its activation appears unlikely to contribute significantly to the developmental pathogenesis of hyperinsulinaemic laminitis. However, inflammation may have a role to play in the later stages (e.g., repair or remodelling) of the disease.

Reply to the comments on “A study of the phosphate mineral kapundaite NaCa(Fe3+)4(PO4)4(OH)3·5(H2O) using SEM/EDX and vibrational spectroscopic methods” by Frost et al. (2014)

"The authors agree with the statements made by Mills and Christy on the study of kapundaite [1]. These authors are correct and have removed any confusion about the origin of the sample kapundaite. The authors (Frost et al.) confirm the sample of kapundaite studied in this work is from the Tom‘s quarry, Australia and can be considered a type material. The authors do not accept the statements by Mills and Christy on “type minerals”. The sample of kapundaite from the Australian source is from the collection of the Geology Department of the Federal University of Ouro Preto, Minas Gerais, Brazil with sample code SAC-111. At least if our mineral sample is not a co-type mineral, our sample is from the same origin as the type mineral. Samples..."--publisher website.

Pro-matrix metalloproteinase-2 transfection increases orthotopic primary growth and experimental metastasis of MDA-MB-231 human breast cancer cells in nude mice

The ability to activate pro-matrix metalloproteinase (pro-MMP)-2 via membrane type-MMP is a hallmark of human breast cancer cell lines that show increased invasiveness, suggesting that MMP-2 contributes to human breast cancer progression. To investigate this, we have stably transfected pro-MMP-2 into the human breast cancer cell line MDA-MB-231, which lacks MMP-2 expression but does express its cell surface activator, membrane type 1-MMP. Multiple clones were derived and shown to produce pro-MMP-2 and to activate it in response to concanavalin A. In vitro analysis showed that the pro-MMP-2-transfected clones exhibited an increased invasive potential in Boyden chamber and Matrigel outgrowth assays, compared with the parental cells or those transfected with vector only. When inoculated into the mammary fat pad of nude mice, each of the MMP-2-tranfected clones grew faster than each of the vector controls tested. After intracardiac inoculation into nude mice, pro-MMP-2-transfected clones showed a significant increase in the incidence of metastasis to brain, liver, bone, and kidney compared with the vector control clones but not lung. Increased tumor burden was seen in the primary site and in lung metastases, and a trend toward increased burden was seen in bone, however, no change was seen in brain, liver, or kidney. This data supports a role for MMP-2 in breast cancer progression, both in the growth of primary tumors and in their spread to distant organs. MMP-2 may be a useful target for breast cancer therapy when refinement of MMP inhibitors provides for MMP-specific agents.

Calibration of the biotrainer pro activity monitor in children

The unique physical and movement characteristics of children necessitate the development of accelerometer equations and cut points that are population specific. The purpose of this study is to develop an ecologically valid cut point for the Biotrainer Pro monitor that reflects a threshold for moderate-intensity physical activity in elementary school children. A sample of 30 children (ages 8-12) wore a Biotrainer monitor while completing a series of 7 movement tasks (calibration phase) and while participating in an organized group activity (cross-validation phase). Videotapes from each session were processed using a computerized direct-observation technique to provide a criterion measure of physical activity. Analyses involved the use of mixed-model regression and receiver operator characteristic (ROC) curves. The results indicated that a cut point of 4 counts/min provides the optimal balance between the related needs for sensitivity (accurately detecting activity) and specificity (limiting misclassification of activity as inactivity). Results with the cross-validation data demonstrated that this value yielded the best overall kappa (.58) and a high classification agreement (84%) for activity determination. The specificity of 93% demonstrates that the proposed cut point can accurately detect activity; however, the lower sensitivity value of 61% suggests that some minutes of activity might be incorrectly classified as inactivity. The cut point of 4 counts/min provides an ecologically valid cut point to capture physical activity in children using the Biotrainer Pro activity monitor.

Selective involvement of TIMP-2 in the second activational cleavage of pro-MMP-2 : refinement of the pro-MMP-2 activation mechanism

A tissue inhibitor of metalloproteinases-2 (TIMP-2)-independent mechanism for generating the first activational cleavage of pro-matrix metalloproteinase-2 (MMP-2) was identified in membrane type-1 MMP (MT1-MMP)-transfected MCF-7 cells and confirmed in TIMP-2-deficient fibroblasts. In contrast, the second MMP-2-activational step was found to be TIMP-2 dependent in both systems. MMP-2 hemopexin C-terminal domain was found to be critical for the first step processing, confirming a need for membrane tethering. We propose that the intermediate species of MMP-2 forms the well-established trimolecular complex (MT1-MMP/TIMP-2/MMP-2) for further TIMP-2-dependent autocatalytic cleavage to the fully active species. This alternate mechanism may supplement the traditional TIMP-2-mediated first step mechanism.

Moving from outsider to insider : peer status and partnerships between electricity utilities and residential consumers

An electricity demand reduction project based on comprehensive residential consumer engagement was established within an Australian community in 2008. By 2011, both the peak demand and grid supplied electricity consumption had decreased to below pre-intervention levels. This case study research explored the relationship developed between the utility, community and individual consumer from the residential customer perspective through qualitative research of 22 residential households. It is proposed that an energy utility can be highly successful at peak demand reduction by becoming a community member and a peer to residential consumers and developing the necessary trust, access, influence and partnership required to create the responsive environment to change. A peer-community approach could provide policymakers with a pathway for implementing pro-environmental behaviour for low carbon communities, as well as peak demand reduction, thereby addressing government emission targets while limiting the cost of living increases from infrastructure expenditure.