Five commercial citrus rate poorly as hosts of the polyphagous fruit flyBactrocera tryoni(Froggatt) (Diptera: Tephritidae) in laboratory studies

Queensland fruit fly, Bactrocera tryoni (Froggatt), is a polyphagous pest, and many citrus types are included among its hosts. While quantification of citrus host use by B. tryoni is lacking, citrus is generally considered a ‘low pressure’ crop. This paper investigates B. tryoni female oviposition preference and offspring performance in five citrus types; Murcott mandarin (Citrus reticulata), Navel orange and Valencia orange (Citrus sinensis), Eureka lemon (Citrus limon) and yellow grapefruit (Citrus paradisi). Oviposition preference was investigated in laboratory-based choice and no-choice experiments, while immature survival and offspring performance were investigated by infesting fruits in the laboratory and evaluating pupal recovery, pupal emergence and F1 fecundity. Fruit size, Brix level and peel toughness were also measured for correlation with host use. Bactrocera tryoni demonstrated an oviposition preference hierarchy among the citrus fruits tested; Murcott and grapefruit were most preferred for oviposition and lemon the least, while preference for Navel and Valencia was intermediate. Peel toughness was negatively correlated with B. tryoni oviposition preference, while no significant correlations were detected between oviposition and Brix level or fruit size. Immature survival in the tested fruit was very low. Murcott was the best host (21% pupal recovery), while all other citrus types that showed pupal recovery of 6% or lower and no pupae were recovered from Valencia orange. In pupae recovered from Navel orange and lemon, adult eclosion was greatly reduced, while in grapefruit and lemon, no eggs were recovered from F1 adults. Based on these laboratory results, many commercial citrus varieties appear to be poor hosts for B. tryoni and may pose a low post-harvest and quarantine risk. These findings need to be confirmed in the field, as they impact on both pre-harvest and post-harvest countermeasures.

Damage assessment in reinforced concrete flexural members using modal strain energy based method

Damage assessment (damage detection, localization and quantification) in structures and appropriate retrofitting will enable the safe and efficient function of the structures. In this context, many Vibration Based Damage Identification Techniques (VBDIT) have emerged with potential for accurate damage assessment. VBDITs have achieved significant research interest in recent years, mainly due to their non-destructive nature and ability to assess inaccessible and invisible damage locations. Damage Index (DI) methods are also vibration based, but they are not based on the structural model. DI methods are fast and inexpensive compared to the model-based methods and have the ability to automate the damage detection process. DI method analyses the change in vibration response of the structure between two states so that the damage can be identified. Extensive research has been carried out to apply the DI method to assess damage in steel structures. Comparatively, there has been very little research interest in the use of DI methods to assess damage in Reinforced Concrete (RC) structures due to the complexity of simulating the predominant damage type, the flexural crack. Flexural cracks in RC beams distribute non- linearly and propagate along all directions. Secondary cracks extend more rapidly along the longitudinal and transverse directions of a RC structure than propagation of existing cracks in the depth direction due to stress distribution caused by the tensile reinforcement. Simplified damage simulation techniques (such as reductions in the modulus or section depth or use of rotational spring elements) that have been extensively used with research on steel structures, cannot be applied to simulate flexural cracks in RC elements. This highlights a big gap in knowledge and as a consequence VBDITs have not been successfully applied to damage assessment in RC structures. This research will address the above gap in knowledge and will develop and apply a modal strain energy based DI method to assess damage in RC flexural members. Firstly, this research evaluated different damage simulation techniques and recommended an appropriate technique to simulate the post cracking behaviour of RC structures. The ABAQUS finite element package was used throughout the study with properly validated material models. The damaged plasticity model was recommended as the method which can correctly simulate the post cracking behaviour of RC structures and was used in the rest of this study. Four different forms of Modal Strain Energy based Damage Indices (MSEDIs) were proposed to improve the damage assessment capability by minimising the numbers and intensities of false alarms. The developed MSEDIs were then used to automate the damage detection process by incorporating programmable algorithms. The developed algorithms have the ability to identify common issues associated with the vibration properties such as mode shifting and phase change. To minimise the effect of noise on the DI calculation process, this research proposed a sequential order of curve fitting technique. Finally, a statistical based damage assessment scheme was proposed to enhance the reliability of the damage assessment results. The proposed techniques were applied to locate damage in RC beams and slabs on girder bridge model to demonstrate their accuracy and efficiency. The outcomes of this research will make a significant contribution to the technical knowledge of VBDIT and will enhance the accuracy of damage assessment in RC structures. The application of the research findings to RC flexural members will enable their safe and efficient performance.

Novel keratins identified by quantitative proteomic analysis as the major cytoskeletal proteins of equine (Equus caballus) hoof lamellar tissue

The dermo-epidermal interface that connects the equine distal phalanx to the cornified hoof wall withstands great biomechanical demands, but is also a region where structural failure often ensues as a result of laminitis. The cytoskeleton in this region maintains cell structure and facilitates intercellular adhesion, making it likely to be involved in laminitis pathogenesis, although it is poorly characterized in the equine hoof lamellae. The objective of the present study was to identify and quantify the cytoskeletal proteins present in the epidermal and dermal lamellae of the equine hoof by proteomic techniques. Protein was extracted from the mid-dorsal epidermal and dermal lamellae from the front feet of 5 Standardbred geldings and 1 Thoroughbred stallion. Mass spectrometry-based spectral counting techniques, PAGE, and immunoblotting were used to identify and quantify cytoskeletal proteins, and indirect immunofluorescence was used for cellular localization of K14 and K124 (where K refers to keratin). Proteins identified by spectral counting analysis included 3 actin microfilament proteins; 30 keratin proteins along with vimentin, desmin, peripherin, internexin, and 2 lamin intermediate filament proteins; and 6 tubulin microtubule proteins. Two novel keratins, K42 and K124, were identified as the most abundant cytoskeletal proteins (22.0 ± 3.2% and 23.3 ± 4.2% of cytoskeletal proteins, respectively) in equine hoof lamellae. Immunoreactivity to K14 was localized to the basal cell layer, and that to K124 was localized to basal and suprabasal cells in the secondary epidermal lamellae. Abundant proteins K124, K42, K14, K5, and α1-actin were identified on 1- and 2-dimensional polyacrylamide gels and aligned with the results of previous studies. Results of the present study provide the first comprehensive analysis of cytoskeletal proteins present in the equine lamellae by using mass spectrometry-based techniques for protein quantification and identification.

Investigation of energy efficient approaches for the energy performance improvement of commercial buildings

Energy efficiency of buildings is attracting significant attention from the research community as the world is moving towards sustainable buildings design. Energy efficient approaches are measures or ways to improve the energy performance and energy efficiency of buildings. This study surveyed various energy-efficient approaches for commercial building and identifies Envelope Thermal Transfer Value (ETTV) and Green applications (Living wall, Green facade and Green roof) as most important and effective methods. An in-depth investigation was carried out on these energy-efficient approaches. It has been found that no ETTV model has been developed for sub-tropical climate of Australia. Moreover, existing ETTV equations developed for other countries do not take roof heat gain into consideration. Furthermore, the relationship of ETTV and different Green applications have not been investigated extensively in any literature, and the energy performance of commercial buildings in the presence of Living wall, Green facade and Green roof has not been investigated in the sub-tropical climate of Australia. The study has been conducted in two phases. First, the study develops the new formulation, coefficient and bench mark value of ETTV in the presence of external shading devices. In the new formulation, roof heat gain has been included in the integrated heat gain model made of ETTV. In the 2nd stage, the study presents the relationship of thermal and energy performance of (a) Living wall and ETTV (b) Green facade and ETTV (c) Combination of Living wall, Green facade and ETTV (d) Combination of Living wall, Green Roof and ETTV in new formulations. Finally, the study demonstrates the amount of energy that can be saved annually from different combinations of Green applications, i.e., Living wall, Green facade; combination of Living wall and Green facade; combination of Living wall and Green roof. The estimations are supported by experimental values obtained from extensive experiments of Living walls and Green roofs.

Energy-pollution nexus for urban buildings

Since the first oil crisis in 1974, economic reasons placed energy saving among the top priorities in most industrialised countries. In the decades that followed, another, equally strong driver for energy saving emerged: climate change caused by anthropogenic emissions, a large fraction of which result from energy generation. Intrinsically linked to energy consumption and its related emissions is another problem: indoor air quality. City dwellers in industrialised nations spend over 90% of their time indoors and exposure to indoor pollutants contributes to ~2.6% of global burden of disease and nearly 2 million premature deaths per year1. Changing climate conditions, together with human expectations of comfortable thermal conditions, elevates building energy requirements for heating, cooling, lighting and the use of other electrical equipment. We believe that these changes elicit a need to understand the nexus between energy consumption and its consequent impact on indoor air quality in urban buildings. In our opinion the key questions are how energy consumption is distributed between different building services, and how the resulting pollution affects indoor air quality. The energy-pollution nexus has clearly been identified in qualitative terms; however the quantification of such a nexus to derive emissions or concentrations per unit energy consumption is still weak, inconclusive and requires forward thinking. Of course, various aspects of energy consumption and indoor air quality have been studied in detail separately, but in-depth, integrated studies of the energy-pollution nexus are hard to come by. We argue that such studies could be instrumental in providing sustainable solutions to maintain the trade-off between the energy efficiency of buildings and acceptable levels of air pollution for healthy living.

Strategies for improved disease resistance in micro-propagated bananas

Bananas (Musa sp) are one of the most important food crops in the world and provide a staple food and source of income in many households especially in Africa. Diseases are a major constraint to production with bunchy top, caused by Banana bunchy top virus (BBTV) generally considered the most important virus disease of bananas worldwide. Of the fungal diseases, Fusarium wilt, caused by the Fusarium oxysporum f.sp cubense (Foc), and black Sigatoka, caused by Mycosphaerella fijiensis, are arguably two of the most important and cause significant yield losses. The low fertility of commercially important banana cultivars has hampered efforts to generate disease resistance using conventional breeding. Possible alternative strategies to generate or increase disease resistance are through genetic engineering or by manipulation of the innate plant defence mechanisms, namely systemic acquired resistance (SAR). The first research component of this thesis describes attempts to generate BBTV-resistant banana plants using a genetic modification approach. The second research component of the thesis focused on the identification of a potential marker gene associated with SAR in banana plants and a comparison of the expression levels of the marker gene in response to biotic and abiotic stresses, and chemical inducers. Previous research at QUT CTCB showed that replication of BBTV DNA components in banana embryogenic cell suspensions (ECS) was abolished following co-bombardment with 1.1mers of mutated BBTV DNA-R. BBTV DNA-R encodes the master replication protein (Rep) and is the only viral protein essential for BBTV replication. In this study, ECS of banana were stably transformed with the same constructs, each containing a different mutation in BBTV DNA-R, namely H41G, Y79F and K187M, to examine the effect on virus replication in stably transformed plants. Cells were also transformed with a construct containing a native BBTV Rep. A total of 16, 16, 11 and five lines of stably transformed banana plants containing the Y79F, H41G, K187M and native Rep constructs, respectively, were generated. Of these, up to nine replicates from Y79F lines, four H41G lines, seven K187M lines and three native Rep lines were inoculated with BBTV by exposure to viruliferous aphids in two separate experiments. At least one replicate from each of the nine Y79F lines developed typical bunchy top symptoms and all tested positive for BBTV using PCR. Of the four H41G lines tested, at least one replicate from three of the lines showed symptoms of bunchy top and tested positive using PCR. However, none of the five replicates of one H41G line (H41G-3) developed symptoms of bunchy top and none of the plants tested positive for BBTV using PCR. Of the seven K187M lines, at least one replicate of all lines except one (K187M-1) developed symptoms of bunchy top and tested positive for BBTV. Importantly, none of the four replicates of line K187M-1 showed symptoms or tested positive for BBTV. At least one replicate from each of the three native Rep lines developed symptoms and tested positive for BBTV. The H41G-3 and K187M-1 lines possibly represent the first transgenic banana plants generated using a mutated Rep strategy. The second research component of this thesis focused on the identification of SAR-associated genes in banana and their expression levels in response to biotic and abiotic stresses and chemical inducers. The impetus for this research was the observation that tissue-cultured (TC) banana plants were more susceptible to Fusarium wilt disease (and possibly bunchy top disease) than plants grown from field-derived suckers, possibly due to decreased levels of SAR gene expression in the former. In this study, the pathogenesis-related protein 1 (PR-1) gene was identified as a potential marker for SAR gene expression in banana. A quantitative real-time PCR assay was developed and optimised in order to determine the expression of PR-1, with polyubiquitin (Ubi-1) found to be the most suitable reference gene to enable relative quantification. The levels of PR-1 expression were subsequently compared in Lady Finger and Cavendish (cv. Williams) banana plants grown under three different environmental conditions, namely in the field, the glass house and in tissue-culture. PR-1 was shown to be expressed in both cultivars growing under different conditions. While PR-1 expression was highest in the field grown bananas and lowest in the TC bananas in Lady Finger cultivar, this was not the case in the Cavendish cultivar with glass house plants exhibiting the lowest PR-1 expression compared with tissue culture and field grown plants. The important outcomes of this work were the establishment of a qPCR-based assay to monitor PR-1 expression levels in banana and a preliminary assessment of the baseline PR-1 expression levels in two banana cultivars under three different growing conditions. After establishing the baseline PR-1 expression levels in Cavendish bananas, a study was done to determine whether PR-1 levels could be increased in these plants by exposure to known banana pathogens and non-pathogens, and a known chemical inducer of SAR. Cavendish banana plants were exposed to pathogenic Foc subtropical race 4 (FocSR4) and non-pathogenic Foc race 1 (Foc1), as well as two putative inducers of resistance, Fusarium lycopersici (Fol) and the chemical, acibenzolar-S-methyl (BION®). Tissue culture bananas were acclimatised under either glass house (TCS) or field (TCH) conditions and treatments were carried out in a randomised complete block design. PR-1 expression was determined using qPCR for both TCS and TCH samples for the period 12-72h post-exposure. Treatment of TCH plants using Foc1 and FocSR4 resulted in 120 and 80 times higher PR-1 expression than baseline levels, respectively. For TCS plants treated with Foc1, PR-1 expression was 30 times higher than baseline levels at 12h post-exposure, while TCS plants treated with FocSR4 showed the highest PR-1 expression (20 times higher than baseline levels) at 72h post-exposure. Interestingly, when TCS plants were treated with Fol there was a marked increase of PR-1 expression at 12 h and 48 h following treatment which was 4 and 8 times higher than the levels observed when TCS plants were treated with Foc1 and FocSR4, respectively. In contrast, when TCH plants were treated with Fol only a slight increase in PR-1 expression was observed at 12 h, which eventually returned to baseline levels. Exposure of both TCS and TCH plants to BION® resulted in no effect on PR-1 expression levels at any time-point. The major outcome of the SAR study was that the glass house acclimatised tissue culture bananas exhibited lower PR-1 gene expression compared to field acclimatised tissue culture plants and the identification of Fol as a good candidate for SAR induction in banana plants exhibiting low PR-1 levels. A number of outcomes that foster understanding of both pathogen-derived and plant innate resistance strategies in order to potentially improve banana resistance to diseases were explored in this study and include identification of potential inducers of systemic acquired resistance and a promising mutated Rep approach for BBTV resistance. The work presented in this thesis is the first report on the generation of potential BBTV resistant bananas using the mutated Rep approach. In addition, this is the first report on the status of SAR in banana grown under different conditions of exposure to the biotic and abiotic environment. Further, a robust qPCR assay for the study of gene expression using banana leaf samples was developed and a potential inducer of SAR in tissue culture bananas identified which could be harnessed to increase resistance in tissue culture bananas.

Designing a continuum of quality external cause of injury information in Queensland : from ambulance to hospital

This study is the first to employ an epidemiological framework to evaluate the ‘fit-for-purpose’ of ICD-10-AM external cause of injury codes, ambulance and hospital clinical documentation for injury surveillance. Importantly, this thesis develops an evidence-based platform to guide future improvements in routine data collections used to inform the design of effective injury prevention strategies. Quantification of the impact of ambulance clinical records on the overall information quality of Queensland hospital morbidity data collections for injury causal information is a unique and notable contribution of this study.

Simultaneous determination by UPLC-ESI-MS of scoparone, capillarisin, rhein, and emodin in rat urine after oral administration of Yin Chen Hao Tang preparation

A simple, sensitive, and validated method was developed for simultaneous determination of scoparone, capillarisin, rhein, and emodin in rat urine by ultra-performance liquid chromatography/electrospray ionization quadruple time-of-flight mass spectrometry (UPLC-MS). The urinary samples were analyzed on an Acquity UPLC BEH C18 1.7 microm 2.1x50 mm column. Scoparone, capillarisin, rhein, and emodin in rat urine were simultaneously analyzed with good separation. The lower limits of detection were 6.0, 9.0, 7.0, and 3.0 ng/mL, and the lower limits of quantification were 20.0, 33.0, 24.0, and 12.0 ng/mL for scoparone, capillarisin, rhein, and emodin, respectively. The intra- and inter-day precisions (RSD) were less than 9%. The intra- and inter-accuracies were found to be in the range of 94.14-104.54% for scoparone, 101.72-107.34% for capillarisin, 95.24-103.59% for rhein, and 101.32-107.82% for emodin at three concentration levels. The absolute recoveries for scoparone, capillarisin, rhein, and emodin were not less than 77.0%. The developed method has been applied to determine scoparone, capillarisin, rhein, and emodin in rat urine after oral administration of Yin Chen Hao Tang preparation, a traditional Chinese medicine formulation widely used in China for treatment of jaundice and liver disorders.

CD4+ tumor infiltrating lymphocytes are prognostic and independent of R-IPI in patients with DLBCL receiving R-CHOP chemo-immunotherapy

Despite the Revised International Prognostic Index's (R-IPI) undoubted utility in diffuse large B-cell lymphoma (DLBCL), significant clinical heterogeneity within R-IPI categories persists. Emerging evidence indicates that circulating host immunity is a robust and R-IPI independent prognosticator, most likely reflecting the immune status of the intratumoral microenvironment. We hypothesized that direct quantification of immunity within lymphomatous tissue would better permit stratification within R-IPI categories. We analyzed 122 newly diagnosed consecutive DLBCL patients treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) chemo-immunotherapy. Median follow-up was 4 years. As expected, the R-IPI was a significant predictor of outcome with 5-year overall survival (OS) 87% for very good, 87% for good, and 51% for poor-risk R-IPI scores (P < 0.001). Consistent with previous reports, systemic immunity also predicted outcome (86% OS for high lymphocyte to monocyte ratio [LMR], versus 63% with low LMR, P = 0.01). Multivariate analysis confirmed LMR as independently prognostic. Flow cytometry on fresh diagnostic lymphoma tissue, identified CD4+ T-cell infiltration as the most significant predictor of outcome with ≥23% infiltration dividing the cohort into high and low risk groups with regard to event-free survival (EFS, P = 0.007) and OS (P = 0.003). EFS and OS were independent of the R-IPI and LMR. Importantly, within very good/good R-IPI patients, CD4+ T-cells still distinguished patients with different 5 year OS (high 96% versus low 63%, P = 0.02). These results illustrate the importance of circulating and local intratumoral immunity in DLBCL treated with R-CHOP.

Damage assessment in beams and plates using variations in modal properties

Damage detection using modal properties is a widely accepted method. However, quantifying such damage using modal properties is still not well established. With this in mind, a research project is presently underway towards the development of a procedure to detect, locate and quantify damage in structural components using the variations in modal properties. A novel vibration based parameter called Vibration based Damage Index is introduced into the damage assessment procedure. This paper presents the early part of the research project which treats flexural members. The proposed procedure is validated using experimental data and/or theoretical techniques and illustrated through application. Outcomes of this research highlight the ability of the proposed procedure to successfully detect, locate and quantify damage in flexural structural components using the modal properties of the first few modes.

Age-related changes in cardiac adenosine receptor expression

Adenosine is an important cardioprotective agent that works via several adenosine receptor (ADOR) subtypes to regulate cardiovascular activity. It is well established that functional responses to adenosine decline with age. What is unclear, though, is whether these changes occur at the receptor, second messenger or translational level. In this study we determined the effect of age on cardiac adenosine receptor expression using the housekeeping gene 18S rRNA versus the adenosine A2B receptor gene as internal controls. Absolute quantification showed that no age-related changes occurred in the expression of 18S rRNA or adenosine A2B receptor internal control genes. Subsequently, relative analysis of the adenosine receptor subtypes using 18S rRNA found a significant age-related reduction in the expression of the adenosine A1 receptor (5.5-fold), with no changes in the expression of the adenosine A2A, A2B and A3 receptors. When using the expression of the adenosine A2B receptor as the internal control gene, a significant down regulation of both the adenosine A1 (5.4-fold) and A2A (2.2-fold) receptors with no change in the expression of adenosine A3 receptor was found. Therefore, the high level of expression of the 18S rRNA housekeeping gene was found to mask a significant change in expression of the adenosine A2A receptor with age. Ultimately, these findings show an age-related reduction in adenosine A1 and A2A receptor expression in rat heart.

Identification and simultaneous analysis of harmane, harmol, isovitexin and vitexin in Passiflora incarnata extracts with a novel HLPC method

A high performance liquid chromatographic method for the simultaneous analysis of two flavonoids (iso-vitexin and vitexin), and three indole alkaloids (harmane, harmine, and harmol) was developed. This method was then utilised to quantitate levels of these five constituents in methanolic extracts of Australian Passiflora incarnata. HPLC analysis was performed using a Waters™ Novapak C18 (150 × 4 mm, 4 μm) column, with a gradient solvent system of methanol-water-acetic acid. Detection was achieved by PDA UV (254 nm) and fluorescence (excitation 254 nm, emission 414 nm), utilising the external standard method to obtain quantification.

Simultaneous determination of aldrin, dieldrin, endrin, heptachlor, and p,p'-DDT in medicinal plant extracts using a novel high performance liquid chromatography method

A high performance liquid chromatographic method for the simultaneous determination of five organochlorine pesticides (aldrin, p,p’-DDT, dieldrin, endrin, and heptachlor) was developed. The method was used to determine the levels of these pesticides in medicinal plant samples. Analysis was carried out using a Merck LiChrospher 100 RP C18 (5 μm) column with a gradient solvent system of acetonitrile-water and PDA UV detection (224 nm). Quantification was carried out by the external standard method. The limit of detection for the utilized method was below the local legal limits (ANZFA) for similar plant materials for all 5 pesticides excepting endrin. Medicinal plant extracts were further analyzed by conventional GC-ECD and GC-NPD means using SPE and GPC cleanup as required.

Accounting for volunteer services : a deficiency in accountability

The purpose of this study is to identify the extent to which NFP organisations disclose information on volunteer contributions of services. Design/methodology/approach – The study relies on information disclosed in the websites of NFP organisations. Findings - We find that disclosure was more prevalent on NFP websites compared to digital annual report disclosures. We find that more NFPs provided disclosure on the activities of their volunteers than other items pertaining to volunteers and the quantification and valuation of volunteer contributions were the least likely to be disclosed. Importantly, the findings illustrate an accountability deficiency in the comprehensiveness of disclosure which results in an under-representation of the contribution volunteers provide to organisational sustainability and impact on mission fulfilment. Research limitations/implications – The convenience sample size restricts further interrogation to tease out organisational characteristics that may influence current disclosure practices. Practical implications - The findings contribute to international debate over the inclusion of volunteer contributions in the assessment of a NFP’s accountability over its resources and ultimately the enhancement of its sustainability.

Characterisation of microvasulature changes after closed soft tissue trauma by contrast enhanced micro-CT imaging

INTRODUCTION It is known that the vascular morphology and functionality are changed following closed soft tissue trauma (CSTT) [1], and bone fractures [2]. The disruption of blood vessels may lead to hypoxia and necrosis. Currently, most clinical methods for the diagnosis and monitoring of CSTT with or without bone fractures are primarily based on qualitative measures or practical experience, making the diagnosis subjective and inaccurate. There is evidence that CSTT and early vascular changes following the injury delay the soft tissue tissue and bone healing [3]. However, a precise qualitative and quantitative morphological assessment of vasculature changes after trauma is currently missing. In this research, we aim to establish a diagnostic framework to assess the 3D vascular morphological changes after standardized CSTT in a rat model qualitatively and quantitatively using contrast-enhanced micro-CT imaging. METHODS An impact device was used for the application of a controlled reproducible CSTT to the left thigh (Biceps Femoris) of anaesthetized male Wistar rats. After euthanizing the animals at 6 hours, 24 hours, 3 days, 7 days, or 14 days after trauma, CSTT was qualitatively evaluated by macroscopic visual observation of the skin and muscles. For visualization of the vasculature, the blood vessels of sacrificed rats were flushed with heparinised saline and then perfused with a radio-opaque contrast agent (Microfil, MV 122, Flowtech, USA) using an infusion pump. After allowing the contrast agent to polymerize overnight, both hind-limbs were dissected, and then the whole injured and contra-lateral control limbs were imaged using a micro-CT scanner (µCT 40, Scanco Medical, Switzerland) to evaluate the vascular morphological changes. Correlated biopsy samples were also taken from the CSTT region of both injured and control legs. The morphological parameters such as the vessel volume ratio (VV/TV), vessel diameter (V.D), spacing (V.Sp), number (V.N), connectivity (V.Conn) and the degree of anisotropy (DA) were then quantified by evaluating the scans of biopsy samples using the micro-CT imaging system. RESULTS AND DISCUSSION A qualitative evaluation of the CSTT has shown that the developed impact protocols were capable of producing a defined and reproducible injury within the region of interest (ROI), resulting in a large hematoma and moderate swelling in both lateral and medial sides of the injured legs. Also, the visualization of the vascular network using 3D images confirmed the ability to perfuse the large vessels and a majority of the microvasculature consistently (Figure 1). Quantification of the vascular morphology obtained from correlated biopsy samples has demonstrated that V.D and V.N and V.Sp were significantly higher in the injured legs 24 hours after impact in comparison with the control legs (p<0.05). The evaluation of the other time points is currently progressing. CONCLUSIONS The findings of this research will contribute to a better understanding of the changes to the vascular network architecture following traumatic injuries and during healing process. When interpreted in context of functional changes, such as tissue oxygenation, this will allow for objective diagnosis and monitoring of CSTT and serve as validation for future non-invasive clinical assessment modalities.