Characterization and novel activation of 72-kDa metalloproteinase in retinal interphotoreceptor matrix and Y-79 cell culture medium

Analysis of bovine interphotoreceptor matrix and conditioned medium from human Y-79 retinoblastoma cells by gelatin SDS-PAGE zymography reveals abundant activity of a 72-kDa M(r) gelatinase. The 72-kDa gelatinase from either source is inhibited by EDTA but not aprotinin or NEM, indicating that it is a metalloproteinase (MMP). The 72-kDa MMP is converted to a 62-kDa species with APMA treatment after gelatin sepharose affinity purification typical of previously described gelatinase MMP-2. The latent 72-kDa gelatinase from either bovine IPM or Y-79 media autoactivates without APMA in the presence of calcium and zinc after 72 hr at 37°C, producing a fully active mixture of proteinase species, 50 (48 in Y-79 medium), 38 and 35 kDa in size. The presence of inhibitory activity was examined in both whole bovine IPM and IPM fractions separated by SDS-PAGE. Whole IPM inhibited gelatinolytic activity of autoactivated Y-79-derived MMP in a dose-dependent manner. Inhibitory activities are observed in two protein fractions of 27-42 and 20-25 kDa. Western blots using antibodies to human tissue inhibitor of metalloproteinase 1 and 2 (TIMP-1 and -2) reveal the presence of two TIMP-1-like proteins at 21 and 29 kDa in inhibitory fractions of the bovine IPM. TIMP-2 was not detected in the inhibitory IPM fractions, consistent with the observed autoactivation of bovine IPM 72-kDa gelatinase. Potential roles for this IPM MMP-TIMP system include physiologic remodelling of the neural retina-RPE cell interface and digestion of shed rod outer segment as well as pathological processes such as retinal detachment, PE cell migration, neovascularization and tumor progression. Cultured Y-79 cells appear to be a good model for studying the production and regulation of this proteinase system.

CFTR expression is regulated during both the cycle of the seminiferous epithelium and the oestrous cycle of rodents

Severely reduced fertility is a common finding in cystic fibrosis (CF). We used in situ hybridization to examine the cell-specific expression of CFTR in the reproductive organs of rodents. In males CFTR mRNA is found in the round spermatids (spermatogenic stages V-X) and in the principal cells that line the initial segment of the epididymis. In both the testis and the epididymis, CFTR expression is developmentally regulated suggesting that the defect in the genital tract of male CF patients is of developmental origin. CFTR expression in the luminal and glandular epithelium of the uterus is regulated during the oestrous cycle and is maximal at pro-oestrus. Our results provide a biological rationale for the reduced fertility of CF patients, and suggest a possible cause for the comparatively poorer prognosis for women with CF.

Graduate careers in journalism, media and communications within and outside the sector : early career outcomes, trajectories and capabilities

This chapter addresses the radical paucity of empirical data about the career destinations of journalism, media and communications graduates from degree programs. We report findings from a study of ten years of graduates from Queensland University of Technology’s courses in journalism, media, and communication studies, using a ‘Creative Trident’ lens to analyse micro individual survey data. The study findings engage with creative labour precarity discussions, and also assertions of creative graduate oversupply suggested by national graduate outcome statistics. We describe the graduates’ employment outcomes, characterise their early career movements into and out of embedded and specialist employment, and compare the capability requirements and degree of course relevance reported by graduates employed in the different Trident segments. Given that in general the graduates in this study enjoyed very positive employment outcomes, but that there were systematic differences in reported course relevance by segment of employment and role, we also consider how university programs can best engage with the task of educating students for a surprisingly diverse range of media and communication-related occupational outcomes within and outside the creative industries.

Segmental torso masses in adolescent idiopathic scoliosis

Background Adolescent Idiopathic Scoliosis is the most common type of spinal deformity whose aetiology remains unclear. Studies suggest that gravitational forces in the standing position play an important role in scoliosis progression, therefore anthropometric data are required to develop biomechanical models of the deformity. Few studies have analysed the trunk by vertebral level and none have performed investigations of the scoliotic trunk. The aim of this study was to determine the centroid, thickness, volume and estimated mass, for sections of the trunk in Adolescent Idiopathic Scoliosis patients. Methods Existing low-dose Computed Tomography scans were used to estimate vertebral level-by-level torso masses for 20 female Adolescent Idiopathic Scoliosis patients. ImageJ processing software was used to analyse the Computed Tomography images and enable estimation of the segmental torso mass corresponding to each vertebral level. Findings The patients’ mean age was 15.0 (SD 2.7) years with mean major Cobb Angle of 52° (SD 5.9) and mean patient weight of 58.2 (SD 11.6) kg. The magnitude of torso segment mass corresponding to each vertebral level increased by 150% from 0.6kg at T1 to 1.5kg at L5. Similarly, the segmental thickness corresponding to each vertebral level from T1-L5 increased inferiorly from a mean 18.5 (SD 2.2) mm at T1 to 32.8 (SD 3.4) mm at L5. The mean total trunk mass, as a percentage of total body mass, was 27.8 (SD 0.5) % which was close to values reported in previous literature. Interpretation This study provides new anthropometric reference data on segmental (vertebral level-by-level) torso mass in Adolescent Idiopathic Scoliosis patients, useful for biomechanical models of scoliosis progression and treatment.

Segmental torso masses and joint torques produced by gravity in the adolescent scoliotic spine

Introduction Calculating segmental torso masses in Adolescent Idiopathic Scoliosis (AIS) patients allows the gravitational loading on the scoliotic spine during relaxed standing to be estimated. Methods Low dose CT data was used to calculate vertebral level-by-level torso masses and spinal joint torques for 20 female AIS patients (mean age 15.0 ± 2.7 years, mean Cobb angle 53 ± 7.1°). ImageJ software (v1.45 NIH USA) was used to threshold the T1 to L5 CT images and calculate the segmental torso volume and mass for each vertebral level. Masses for the head, neck and arms were taken from published data. Intervertebral joint torques in the coronal and sagittal planes at each vertebral level were found from the position of the centroid of the segment masses relative to the joint centres (assumed to be at the centre of the intervertebral disc. The joint torque at each level was found by summing torque contributions for all segments above that joint. Results Segmental torso mass increased from 0.6kg at T1 to 1.5kg at L5. The coronal plane joint torques due to gravity were 5-7Nm at the apex of the curve; sagittal torques were 3-5.4Nm. Conclusion CT scans were in the supine position and curve magnitudes are known to be smaller than those in standing. Hence, this study has shown that gravity produces joint torques potentially of higher than 7Nm in the coronal plane and 5Nm in the sagittal plane during relaxed standing in scoliosis patients. The magnitude of these torques may help to explain the mechanics of AIS progression and the mechanics of bracing. This new data on torso segmental mass in AIS patients will assist biomechanical models of scoliosis.

Caring for emergency service personnel : does what we do work?

Most emergency service organisations have some form of staff support program that share general aims of promoting and maintaining the mental health of their workforce. Yet few of these services have been subject to evaluation and fewer still have commissioned external professional researchers to scrutinise their programs. The Queensland Ambulance (QAS) Service provides a comprehensive and multifaceted program that is both proactive and reactive in design and with the support of the Commissioner, was the subject of a rigorous evaluation throughout 2013. In this paper the program services are briefly outlined and the considered approach to the evaluation is presented within the context of existing scientific literature. Using focus groups, information regarding the uptake of the program’s various ‘arms’, and survey data, results suggest the program is widely used and that staff are very satisfied with the services provided. Further, analysis of established psychometric measures demonstrated organisational and interpersonal factors that are important in the promotion of mental health and in warding off the deleterious impacts that frontline emergency service staff can endure. Data presented in this paper indicate how best to ensure a professional quality of life for ambulance personnel, how to promote resilience to the sometimes extremely challenging aspects of the work role, and ways in which difficulties such as depression may be minimised.

Promoting posttraumatic growth in police recruits : preliminary results of a randomised controlled resilience intervention trial

Recent research has demonstrated that the same experiences that may elicit symptoms of post-traumatic stress disorder (PTSD) in emergency service personnel can also provide a catalyst for positive personal changes such as posttraumatic growth (PTG). In this research newly recruited police officers (N = 412) participated in a randomised control trial of a program specifically designed to promote mental health. On entry to the academy, new recruits were randomly allocated, by classrooms, to either a treatment as usual condition (i.e., existing psychoeducation program) or to the intervention group. The Promoting Resilient Officers (PRO) program is a resilience building intervention adapted from an earlier resilience building program in collaboration with the police service. The PRO program also includes additional components on trauma and PTG. The current research included the participants who had experienced trauma prior to or during the research period (N = 246). It was hypothesised that participation in the PRO program would increase levels of PTG and lower levels of PTSD when compared to recruits in the control condition. Using multilevel modelling and post-hoc analyses, results indicated there were significantly higher levels of PTG across multiple dimensions when compared to the control group. There was no effect on PTSD symptoms with both conditions showing a floor effect. The research indicated the potential value of developing interventions that elicit reflections on the potential for positive as well as negative outcomes of experiencing traumatic and other highly challenging events.

Challenges to ES Success : A Critical Need for Knowledge Integration

Recent literature on Enterprise System (ES) implementation projects highlights the importance of Knowledge Integration (KI) for implementation success. The fundamental characteristics of ES - integration of modules, business process view, and aspects of information transparency - necessitate that all frequent end-users share a reasonable amount of common knowledge and integrate their knowledge to yield new knowledge. Unfortunately, the importance of KI is often overlooked and little about the role of KI in ES success is known. In this chapter, the authors study the KI impact on ES success that is relevant to the ES post-implementation in support of organizations' returns on their ES investments. They adopt the ES post-implementation segment of ES utilization to explore whether the KI approach is causally linked to ES success. The research model was tested in a multi-industry sample in Malaysia from which data was gathered from managerial and operational employees spread across six large organizations. Consistent with the explanation by knowledge-based theory, the results show that KI was valid and significantly related to the outcome of ES that relates to an organization's performance, which the authors refer to as ES success. The KI positive impact on the success of ES drives one to highlight the importance of ontological KI in the complexity of the ES environment. The authors believe that focusing on an ontology through the KI perspective can make significant contributions to current ES problems.

A biomechanical analysis of growing rods used in the management of early onset scoliosis

INTRODUCTION Managing spinal deformities in young children is challenging, particularly early onset scoliosis (EOS). Surgical intervention is often required if EOS has been unresponsive to conservative treatment particularly with rapidly progressive curves. An emerging treatment option for EOS is fusionless scoliosis surgery. Similar to bracing, this surgical option potentially harnesses growth, motion and function of the spine along with correcting spinal deformity. Dual growing rods are one such fusionless treatment, which aims to modulate growth of the vertebrae. The aim of this study was to ascertain the extent to which semiconstrained growing rods (Medtronic, Sofamor, Danek, Memphis, TN) with a telescopic sleeve component, reduce rotational constraint on the spine compared with standard "constrained / rigid" rods and hence potentially provide a more physiological mechanical environment for the growing spine. METHODS Six 40-60kg English Large White porcine spines served as a model for the paediatric human spine. Each spine was dissected into a 7 level thoracolumbar multi-segment unit (MSU), removing all non-ligamentous soft tissues and leaving 3cm of ribs either side. Pure nondestructive axial rotation moments of ±4Nm at a constant rotation rate of 8deg.s-1 were applied to the mounted MSU spines using a biaxial Instron testing machine. Displacement of each vertebral level was captured using a 3D motion tracking system (Optotrak 3020, Northern Digital Inc, Waterloo, ON). Each spine was tested in an un-instrumented state first and then with appropriately sized semi-constrained growing rods and rigid rods in alternating sequence. The rods were secured by multi-axial pedicle screws (Medtronic CD Horizon) at levels 2 and 6 of the construct. The range of motion (ROM), neutral zone (NZ) size and stiffness (Nm.deg-1) were calculated from the Instron load-displacement data and intervertebral ROM was calculated through a MATLAB algorithm from Optotrak data. RESULTS Irrespective of the order of testing, rigid rods significantly reduced the total ROM compared with semi-constrained rods (p<0.05) with in a significantly stiffer spine for both left and right axial rotation (p<0.05). Analysing the intervertebral motion within the instrumented levels 2-6, rigid rods showed reduced ROM compared with semi-constrained growing rods and compared with un-instrumented motion segments. CONCLUSION Semi-constrained growing rods maintain similar stiffness in axial rotation to un-instrumented spines, while dual rigid rods significantly reduce axial rotation. Clinically the effect of semi-constrained growing rods as observed in this study is that they would be expected to allow growth via the telescopic rod components while maintaining the axial flexibility of the spine, which may reduce occurrence of the crankshaft phenomenon.

The stabilising role of the costovertebral joints and spinous processes on thoracic spine stability

Introduction The importance of in vitro biomechanical testing in today’s understanding of spinal pathology and treatment modalities cannot be stressed enough. Different studies have used differing levels of dissection of their spinal segments for their testing protocols[1, 2]. The aim of this study was to assess the impact of removing the costovertebral joints and partial resection of the spinous process sequentially, on the stiffness of the immature thoracic bovine spinal segment. Materials and Methods Thoracic spines from 6-8 week old calves were used. Each spine was dissected and divided into motion segments with 5cm of attached rib on each side and full spinous processes including levels T4-T11 (n=28). They were potted in polymethylemethacrylate. An Instron Biaxial materials testing machine with a custom made jig was used for testing. The segments were tested in flexion/extension, lateral bending and axial rotation at 37⁰C and 100% humidity, using moment control to a maximum 1.75 Nm with a loading rate of 0.3 Nm per second. They were first tested intact for ten load cycles with data collected from the tenth cycle. Progressive dissection was performed by removing first the attached ribs, followed by the spinous process at its base. Biomechanical testing was carried out after each level of dissection using the same protocol. Statistical analysis of the data was performed using repeated measures ANOVA. Results In combined flexion/extension there was a significant reduction in stiffness of 16% (p=0.002). This was mainly after resection of the ribs (14%, p=0.024) and mainly occurred in flexion where stiffness reduced by 22% (p=0.021). In extension, stiffness dropped by 13% (p=0.133). However there was no further significant change in stiffness on resection of the spinous process (<1%) (p=1.00). In lateral bending there was a significant decrease in stiffness of 13% (p<0.001). This comprised a drop of 11% on resection of the ribs (p=0.009) and a further 8% on resection of the spinous process (p=0.014). There was no difference between left and right bending. In axial rotation there was no significant change in stiffness after each stage of dissection (p=0.253). There was no difference between left and right rotation. Conclusion The costovertebral joints play a significant role in providing stability to the bovine thoracic spine in both flexion/extension and lateral bending, whereas the spinous processes play a minor role. Both elements have little effect on axial rotation stability.

Intervertebral staple grading system with micro-CT

Introduction Intervertebral stapling is a leading method of fusionless scoliosis treatment which attempts to control growth by applying pressure to the convex side of a scoliotic curve in accordance with the Hueter-Volkmann principle. In addition to that, staples have the potential to damage surrounding bone during insertion and subsequent loading. The aim of this study was to assess the extent of bony structural damage including epiphyseal injury as a result of intervertebral stapling using an in vitro bovine model. Materials and Methods Thoracic spines from 6-8 week old calves were dissected and divided into motion segments including levels T4-T11 (n=14). Each segment was potted in polymethylemethacrylate. An Instron Biaxial materials testing machine with a custom made jig was used for testing. The segments were tested in flexion/extension, lateral bending and axial rotation at 37⁰C and 100% humidity, using moment control to a maximum 1.75 Nm with a loading rate of 0.3 Nm per second for 10 cycles. The segments were initially tested uninstrumented with data collected from the tenth load cycle. Next an anterolateral 4-prong Shape Memory Alloy (SMA) staple (Medtronic Sofamor Danek, USA) was inserted into each segment. Biomechanical testing was repeated as before. The staples were cut in half with a diamond saw and carefully removed. Micro-CT scans were performed and sagittal, transverse and coronal reformatted images were produced using ImageJ (NIH, USA).The specimens were divided into 3 grades (0, 1 and 2) according to the number of epiphyses damaged by the staple prongs. Results: There were 9 (65%) segments with grade 1 staple insertions and 5 (35%) segments with grade 2 insertions. There were no grade 0 staples. Grade 2 spines had a higher stiffness level than grade 1 spines, in all axes of movement, by 28% (p=0.004). This was most noted in flexion/extension with an increase of 49% (p=0.042), followed by non-significant change in lateral bending 19% (p=0.129) and axial rotation 8% (p=0.456) stiffness. The cross sectional area of bone destruction from the prongs was only 0.4% larger in the grade 2 group compared to the grade 1 group (p=0.961). Conclusion Intervertebral staples cause epiphyseal damage. There is a difference in stiffness between grade 1 and grade 2 staple insertion segments in flexion/extension only. There is no difference in the cross section of bone destruction as a result of prong insertion and segment motion.

Gravity-induced coronal plane joint moments in the adolescent scoliotic spine

INTRODUCTION Calculating segmental (vertebral level-by-level) torso masses in Adolescent Idiopathic Scoliosis (AIS) patients allows the gravitational loading on the scoliotic spine during relaxed standing to be estimated. METHODS Existing low dose CT scans were used to calculate vertebral level-by-level torso masses and joint moments occurring in the spine for a group of female AIS patients with right-sided thoracic curves. Image processing software, ImageJ (v1.45 NIH USA) was used to reconstruct the torso segments and subsequently measure the torso volume and mass corresponding to each vertebral level. Body segment masses for the head, neck and arms were taken from published anthropometric data. Intervertebral joint moments at each vertebral level were found by summing each of the torso segment masses above the required joint and multiplying it by the perpendicular distance to the centre of the disc. RESULTS AND DISCUSSION Twenty patients were included in this study with a mean age of 15.0±2.7 years and a mean Cobb angle 52±5.9°. The mean total trunk mass, as a percentage of total body mass, was 27.8 (SD 0.5) %. Mean segmental torso mass increased inferiorly from 0.6kg at T1 to 1.5kg at L5. The coronal plane joint moments during relaxed standing were typically 5-7Nm at the apex of the curve (Figure 1), with the highest apex joint of 7Nm. CT scans were performed in the supine position and curve magnitudes are known to be 7-10° smaller than those measured in standing [1]. Therefore joint moments produced by gravity will be greater than those calculated here. CONCLUSIONS Coronal plane joint moments as high as 7Nm can occur during relaxed standing in scoliosis patients, which may help to explain the mechanics of AIS progression. The body mass distributions calculated in this study can be used to estimate joint moments derived using other imaging modalities such as MRI and subsequently determine if a relationship exists between joint moments and progressive vertebral deformity.

A biomechanical analysis of growing rods used in the management of early onset scoliosis

INTRODUCTION Managing spinal deformities in young children is challenging, particularly early-onset scoliosis (EOS). Any progressive spinal deformity particularly in early life presents significant health risks for the child and a challenge for the treating surgeon. Surgical intervention is often required if EOS has been unresponsive to conservative treatment particularly with rapidly progressive curves. An emerging treatment option particularly for EOS is fusionless scoliosis surgery. Similar to bracing this surgical option potentially harnesses growth, motion and function of the spine along with correcting spinal deformity. Dual growing rods is one such fusionless treatment, which aims to modulate growth of the vertebrae. The aim of this study was to ascertain the extent to which semi-constrained growing rods (Medtronic, Memphis, TN) with a telescopic sleeve component, reduce rotational constraint on the spine compared with standard rigid rods and hence potentially provide a more physiological mechanical environment for the growing spine. METHODS Six 40-60kg English Large White porcine spines served as a model for the paediatric human spine. Each spine was dissected into 7 level thoracolumbar multi-segment unit (MSU) spines, removing all non-ligamentous soft tissues. Appropriately sized semi-constrained growing rods and rigid rods were secured by multi-axial screws (Medtronic) prior to testing in alternating sequences for each spine. Pure nondestructive moments of +/4Nm at a constant rotation rate of 8deg/s was applied to the mounted MSU spines. Displacement of each level was captured using an Optotrak (Northern Digital Inc, Waterloo, ON). The range of motion (ROM), neutral zone (NZ) size and stiffness (Nm/deg) were calculated from the Instron load-displacement data and intervertebral ROM was calculated through a MATLAB algorithm from Optotrak data. RESULTS Irrespective of sequence order rigid rods significantly reduced the total ROM (deg) than compared to semi-constrained rods (p<0.05) and resulted in a significantly stiffer (Nm/deg) spine for both left and right axial rotation testing (p<0.05). Analysing the intervertebral motion within the instrumented levels, rigid rods showed reduced ROM (Deg) than compared to semi-constrained growing rods and the un-instrumented (UN-IN) test sequences. CONCLUSION The semi-constrained growing rods maintained rotation similar to UN-IN spines while the rigid rods showed significantly reduced axial rotation across all instrumented levels. Clinically the effect of semi-constrained growing rods evaluated in this study is that they will allow growth via the telescopic rod components while maintaining the axial rotation ability of the spine, which may also reduce the occurrence of the crankshaft phenomenon.

Drosophila 3′ UTRs are more complex than protein-coding sequences

The 3′ UTRs of eukaryotic genes participate in a variety of post-transcriptional (and some transcriptional) regulatory interactions. Some of these interactions are well characterised, but an undetermined number remain to be discovered. While some regulatory sequences in 3′ UTRs may be conserved over long evolutionary time scales, others may have only ephemeral functional significance as regulatory profiles respond to changing selective pressures. Here we propose a sensitive segmentation methodology for investigating patterns of composition and conservation in 3′ UTRs based on comparison of closely related species. We describe encodings of pairwise and three-way alignments integrating information about conservation, GC content and transition/transversion ratios and apply the method to three closely related Drosophila species: D. melanogaster, D. simulans and D. yakuba. Incorporating multiple data types greatly increased the number of segment classes identified compared to similar methods based on conservation or GC content alone. We propose that the number of segments and number of types of segment identified by the method can be used as proxies for functional complexity. Our main finding is that the number of segments and segment classes identified in 3′ UTRs is greater than in the same length of protein-coding sequence, suggesting greater functional complexity in 3′ UTRs. There is thus a need for sustained and extensive efforts by bioinformaticians to delineate functional elements in this important genomic fraction. C code, data and results are available upon request.

Modelling total duration of traffic incidents including incident detection and recovery time

Traffic incidents are key contributors to non-recurrent congestion, potentially generating significant delay. Factors that influence the duration of incidents are important to understand so that effective mitigation strategies can be implemented. To identify and quantify the effects of influential factors, a methodology for studying total incident duration based on historical data from an ‘integrated database’ is proposed. Incident duration models are developed using a selected freeway segment in the Southeast Queensland, Australia network. The models include incident detection and recovery time as components of incident duration. A hazard-based duration modelling approach is applied to model incident duration as a function of a variety of factors that influence traffic incident duration. Parametric accelerated failure time survival models are developed to capture heterogeneity as a function of explanatory variables, with both fixed and random parameters specifications. The analysis reveals that factors affecting incident duration include incident characteristics (severity, type, injury, medical requirements, etc.), infrastructure characteristics (roadway shoulder availability), time of day, and traffic characteristics. The results indicate that event type durations are uniquely different, thus requiring different responses to effectively clear them. Furthermore, the results highlight the presence of unobserved incident duration heterogeneity as captured by the random parameter models, suggesting that additional factors need to be considered in future modelling efforts.