PPARγ ligands, 15-deoxy-Δ12,14-prostaglandin J2 and rosiglitazone regulate human cultured airway smooth muscle proliferation through different mechanisms

1. The influence of two peroxisome proliferator-activated receptor γ (PPARγ) ligands, a thiazolidinedione, rosiglitazone (RG) and the prostaglandin D2 metabolite 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) on the proliferation of human cultured airway smooth muscle (HASM) was examined.

2. The increases in HASM cell number in response to basic fibroblast growth factor (bFGF, 300 pm) or thrombin (0.3 U ml−1) were significantly inhibited by either RG (1–10 μm) or 15d-PGJ2 (1–10 μm). The effects of RG, but not 15d-PGJ2, were reversed by the selective PPARγ antagonist GW9662 (1 μm).

3. Neither RG nor 15d-PGJ2 (10 μm) decreased cell viability, or induced apoptosis, suggesting that the regulation of cell number was due to inhibition of proliferation, rather than increased cell death.

4. Flow-cytometric analysis of HASM cell cycle distribution 24 h after bFGF addition showed that RG prevented the progression of cells from G1 to S phase. In contrast, 15d-PGJ2 caused an increase in the proportion of cells in S phase, and a decrease in G2/M, compared to bFGF alone.

5. Neither RG nor 15d-PGJ2 inhibited ERK phosphorylation measured 6 h post mitogen addition. The bFGF-mediated increase in cyclin D1 protein levels after 8 h was reduced in the presence of 15d-PGJ2, but not RG.

6. Although both RG and 15d-PGJ2 can inhibit proliferation of HASM irrespective of the mitogen used, only the antiproliferative effects of RG appear to be PPARγ-dependent. The different antimitogenic mechanisms of 15d-PGJ2 and synthetic ligands for PPARγ may be exploited to optimise the potential for these compounds to inhibit airway remodelling in asthma.

Contribution of fibroblast and mast cell (afferent) and tumor (efferent) IL-6 effects within the tumor microenvironment

Hyperactive inflammatory responses following cancer initiation have led to cancer being described as a ‘wound that never heals’. These inflammatory responses elicit signals via NFκB leading to IL-6 production, and IL-6 in turn has been shown to induce epithelial to mesenchymal transition in breast cancer cells in vitro, implicating a role for this cytokine in cancer cell invasion. We previously have shown that conditioned medium derived from cancer-associated fibroblasts induced an Epithelial to Mesenchymal transition (EMT) in PMC42-LA breast cancer cells and we have now identify IL-6 as present in this medium. We further show that IL-6 is expressed approximately 100 fold higher in a cancer-associated fibroblast line compared to normal fibroblasts. Comparison of mouse-specific (stroma) and human-specific (tumor) IL-6 mRNA expression from MCF-7, MDA MB 468 and MDA MB 231 xenografts also indicated the stroma rather than tumor as a significantly higher source of IL-6 expression. Mast cells (MCs) feature in inflammatory cancer-associated stroma, and activated MCs secrete IL-6. We observed a higher MC index (average number of mast cells per xenograft section/average tumor size) in MDA MB 468 compared to MDA MB 231 xenografts, where all MC were observed to be active (degranulating). This higher MC index correlated with greater mouse-specific IL-6 expression in the MDA MB 468 xenografts, implicating MC as an important source of stromal IL-6. Furthermore, immunohistochemistry on these xenografts for pSTAT3, which lies downstream of the IL-6 receptor indicated frequent correlations between pSTAT3 and mast cell positive cells. Analysis of publically available databases for IL-6 expression in patient tissue revealed higher IL-6 in laser capture microdissected stroma compared to adjacent tissue epithelium from patients with inflammatory breast cancer (IBC) and invasive non-inflammatory breast cancer (non-IBC) and we show that IL-6 expression was significantly higher in Basal versus Luminal molecular/phenotypic groupings of breast cancer cell lines. Finally, we discuss how afferent and efferent IL-6 pathways may participate in a positive feedback cycle to dictate tumor progression.

Mitomycin C: a promising agent for the treatment of canine corneal scarring

Objective  To evaluate the safety and efficacy of mitomycin C (MMC) in prevention of canine corneal scarring.

Methods  With an in vitro approach using healthy canine corneas, cultures of primary canine corneal fibroblasts or myofibroblasts were generated. Primary canine corneal fibroblasts were obtained by growing corneal buttons in minimal essential medium supplemented with 10% fetal bovine serum. Canine corneal myofibroblasts were produced by growing cultures in serum-free medium containing transforming growth factor β1 (1 ng/mL). Trypan blue assay and phase-contrast microscopy were used to evaluate the toxicity of three doses of MMC (0.002%, 0.02% and 0.04%). Real-time PCR, immunoblot, and immunocytochemistry techniques were used to determine MMC efficacy to inhibit markers of canine corneal scarring.

Results  A single 2-min treatment of 0.02% or less MMC did not alter canine corneal fibroblast or keratocyte phenotype, viability, or growth. The 0.02% dose substantially reduced myofibroblast formation (up to 67%; P < 0.001), as measured by the change in RNA and protein expression of fibrosis biomarkers (α-smooth muscle actin and F-actin).

Conclusion  This in vitro study suggests that a single 2-min 0.02% MMC treatment to the canine corneal keratocytes is safe and may be useful in decreasing canine corneal fibrous metaplasia. In vivo studies are warranted.

A study of seed viability following consumption by birds

This project aimed to quantify the potential for wild birds to spread genetically modified seeds from their area of planting to adjacent areas, through ingestion and transport in the gut (endozoochorous transmission). We addressed this by assessing whether seed viability was affected by passage through the avian gut. We sought to address this question by testing the effects of ingestion of a number of key agricultural crops by three common wild bird pest species.

Examining the viability of windows impact/update method for delay analysis in construction disputes

Nowadays, construction delay disputes often end up on the arbitration route where the delay experts appointed by the parties advise the tribunal on the extension of times entitlements of the parties. For this purpose, the identification and quantification of concurrent and pacing delays are integral aspects of resolving these disputes using a proper delay analysis methodology. The aim of the study is therefore, threefold. Firstly, the available literature on the concurrent and pacing delays are analyzed in detail to establish the principles for the evaluation of the concurrency and pacing delays. Secondly, a robust delay analysis methodology called ‘windows impact/update method’ is explained often used by the experts for the effective quantification of concurrent and pacing delays. This methodology is an improved version of time impact analysis and normal windows analysis. For better demonstration, the explanation of the methodology is facilitated with the help of a typical case study analysis. Finally, the principles of concurrency and pacing, as explained in the literature review, are promptly applied to the case study results to show the applicability of the analysis method on any types of delay disputes. The study shows the effectiveness of the windows impact/update method for the quantification of the concurrent and pacing delays.

Effects of rising temperature on the viability of an important sea turtle rookery

A warming world poses challenges for species with temperature-dependent sex determination, including sea turtles, for which warmer incubation temperatures produce female hatchlings. We combined in situ sand temperature measurements with air temperature records since 1850 and predicted warming scenarios from the Intergovernmental Panel on Climate Change to derive 250-year time series of incubation temperatures, hatchling sex ratios, and operational sex ratios for one of the largest sea turtles rookeries globally (Cape Verde Islands, Atlantic). We estimate that light-coloured beaches currently produce 70.10% females whereas dark-coloured beaches produce 93.46% females. Despite increasingly female skewed sex ratios, entire feminization of this population is not imminent. Rising temperatures increase the number of breeding females and hence the natural rate of population growth. Predicting climate warming impacts across hatchlings, male-female breeding ratios and nesting numbers provides a holistic approach to assessing the conservation concerns for sea turtles in a warming world. © 2014 Macmillan Publishers Limited.

Viability of tallow inclusion in Atlantic salmon diet, as assessed by an on-farm grow out trial

The results of a recent increase in research interest directed at the inclusion of tallow in fish feed formulations are suggesting tallow is viable as a potential substitute for other alternative lipid sources such as poultry by-product oil. Although strong growth performance data has been shown, reservations still exist regarding reduced digestibility and the potential impacts this could have on performance over the duration of a grow-out period in low temperature conditions. Also little information is yet available on the potential effect of dietary tallow inclusion on final product quality. A large scale farm based study testing the inclusion of tallow at 40% inclusion, partially replacing poultry by-product oil, in commercial diets of Atlantic salmon over a winter grow-out period in southern Tasmania, Australia was conducted. Tallow inclusion had no impact on growth performance or nutrient digestibility. Tallow resulted in a slight improvement in fillet quality exhibiting a significant reduction in n - 6 PUFA and the n6:n3 ratio, and an increased n - 3LC-PUFA tissue deposition. Consumers were unable to display any preference in liking between 3 salmon products (cold smoked, hot smoked, and cooked) as a result of tallow inclusion. This study demonstrates the viability of partial inclusion of tallow in Atlantic salmon diets over a winter grow-out period. Statement of relevance: Improved knowledge of alternative dietary energy sources (oils and fats) to be used in aquafeed, (replacing the increasingly expensive, and diminishingly available, fish oil) is a key area of research towards improved environmental sustainability and economic viability of the aquaculture sector. Following a promising laboratory based, research scale, in vivo trial aimed at assessing the viability of tallow in salmon feed, a larger and longer duration farm-based trial was implemented to validate initial findings. Consumer test of final products (fresh-cooked, hot smoked and cold smoked fillets) showed no modification of sensorial attributes. Tallow is hereto shown to be a highly viable alternative oil for the salmon aquafeed industry.

Rent dissipation, political viability and the strategic adoption of free trade agreements

This paper studies the political viability of free trade agreements (FTAs). The key element of the analysis is the “rent dissipation” that these arrangements induce: by eliminating intra-bloc trade barriers, an FTA reduces the incentives of the local firms to lobby for higher external tariffs, thereby causing a reduction of the rents created in the lobbying process. The prospect of rent dissipation moderates the governments’ willingness to participate in FTAs; they will support only arrangements that are “substantially” welfare improving, and no FTA that reduces welfare. Rent dissipation also implies that the prospects of political turnover may create strategic reasons for the formation of FTAs. Specifically, a government facing a high enough probability of losing power may want to form a trade bloc simply to “tie the hands” of its successor. An FTA can affect the likelihood of political turnover as well. If the incumbent party has a known bias toward special interests, it may want to commit to less distortionary policies in order to reduce its electoral disadvantage; the rent dissipation effect ensures that an FTA can serve as the vehicle for such a commitment. In nascent/unstable democracies, the incumbent government can use a free trade agreement also to reduce the likelihood of a dictatorial takeover and to “consolidate” democracy – a finding that is consistent with the timing of numerous accessions to and formations of preferential arrangements.

Effects of the basic fibroblast growth factor and its anti-factor in the healing and collagen maturation of infected skin wound

PURPOSE: The infection is one of the main factors that affect the physiological evolution of the surgical wounds. The aim of this work is to evaluate the effects of fibroblast growth factor (FGFâ) and anti-FGFâ in the healing, synthesis and maturation of collagen when topically used on infected skin wounds of rats. METHODS: An experimental study was perfomed in 60 male Wistar rats. All animals were divided in two groups (A and B). Each group was divided in three subgroups A1, B1; A2, B2 and A3, B3. After anesthesia with pentobarbital, two open squared wounds (1cm2), 4cm distant to each other, were done in the dorsal skin of all the rats. In group A (n=30) the wounds were contaminated with multibacterial standard solution, and in group B(n=30) the wounds were maintained sterile. These wounds were named F1 (for inflammation analysis) and F2 (for collagen study). The open wounds of A1 and B1 rats were topically treated with saline solution, A2 and B2 were treated with FGFâ and subgroups A3 and B3 were treated with FGFâ and anti-FGFâ. The rats were observed until complete epitelization of F2 wounds for determination of healing time and the expression of types I and III collagen, using Picro Sirius Red staining. Inflammatory reaction in F1 wounds was studied using hematoxilineosin staining. The three variable was measured by the Image Pro-Plus Média Cybernetics software. The statistical analysis was performed by ANOVA and Tukey test, considering p<0.05 as significant. RESULTS: It was observed that infection retarded significantly (p<0.05) the time of wound scarring and the topical application of FCFb reverted the inhibition of healing caused by bacteria. The inflammatory reaction was greater in the subgroup B2 than in B1 and A3, and the difference was significant (p<0.05). It was observed greater expression of type I collagen in all the subgroups treated with FCFb, when compared with the untreated subgroups. Type III collagen was significantly decreased in wounds of B3 rats, comparing to the other subgroups. CONCLUSIONS: The FCFb accelerated the healing of open infected wounds and contributed with maturation of collagen, enhancing the type I collagen density. The anti-FCFb antibody was able to attenuate the production of both type I and III collagen