Secreted products and extracellular matrix from testicular peritubular myoid cells influence androgen-binding protein secretion by Sertoli cells in culture

Metabolic cooperation mediated by secreted factors between Sertoli cells and peritubular myoid cells has been well documented. We have confirmed that factors secreted by peritubular myoid cells modulate androgen-binding protein (ABP) secretion by Sertoli cells and shown further that this can also be achieved with peritubular myoid cell extracellular matrix (ECM). While peritubular myoid cell ECM potentiated the stimulatory effect of dibutyryl cyclic AMP on Sertoli cell ABP secretion, secreted factors did not, suggesting that the two components influence Sertoli cells through distinct mechanisms. We also tested other factors and other cell lines for effects on ABP production by Sertoli cells. The addition of human plasma fibronectin or conditioned medium from the basement membrane-producing Englebreth-Holm- Swarm sarcoma also stimulated ABP secretion by Sertoli cells. Cocultures of epithelial Sertoli cells with the cells of mesenchymal origin, such as testicular peritubular myoid cells, embryonic skin fibroblasts, and bladder smooth muscle cells, significantly stimulated ABP secretion by Sertoli cells, but co-culture with the epithelial-derived Martin-Darby canine kidney cell line had no effect on Sertoli cell-secreted ABP levels. Our data further define the epithelial-mesenchymal cell interaction that exists between Sertoli cells and peritubular myoid cells in the mammalian testis.

Sertoli cells as paracrine modulators of DNA synthesis in rat peritubular myoid cells in culture

To determine whether Sertoli cells influence DNA synthesis by rat peritubular myoid cells in vitro, the effects of Sertoli cells on [3H]thymidine incorporation by peritubular myoid cells in a coculture situation were examined. Incubation of testicular peritubular myoid cells with Sertoli cells in coculture induced a significant increase in [3H]thymidine incorporation by peritubular myoid cells. This indicates a cell-cell cooperation between Sertoli and peritubular myoid cells in the testis in terms of DNA synthesis. Secreted factors from Sertoli cells, as tested in a parabiotic culture situation, also increased [3H]thymidine incorporation by peritubular myoid cells. Moreover, in terms of total cellular protein, cocultures of Sertoli cells and peritubular myoid cells resulted in a significant increase when compared with the monocultures, and this coculture effect substituted for the stimulatory response of serum on peritubular myoid cell monoculture. This study investigated the cooperative role of Sertoli cells and peritubular myoid cells in paracrine regulation of testicular functions.

Localization of fibronectin in the rat testis : effects of serum and dibutyryl cyclic AMP on fibronectin deposition by peritubular myoid cells in culture

The peritubular zone of the rat testis has an extensive extracellular matrix (ECM). Fibronectin (FN) is distributed primarily in the basal lamina of the seminiferous tubule boundary tissue and is synthesized by peritubular myoid cells. Several extracellular changes are mediated by growth factors and these changes occur at the time of hormone mediated testicular development, particularly in the peritubular zone. The effects of serum or dibutyryl cyclic AMP (cAMP) on FN production by the mesenchymal peritubular myoid cells were evaluated. Rats of various ages (10, 15, 20, 40 and 80 days) were employed for immunofluorescent localization of rat testicular FN in frozen sections. In all age groups tested, FN was primarily present in a broad layer around each seminiferous tubule, and blood vessel, and in variable distribution throughout the interstitial stroma. By day 20 there was no clear distinction in FN staining between the peritubular zone and the interstitial tissue. This indicates an involvement of FN in the ECM developments which occur in the peritubular zone of the testis at this time. The peritubular myoid cells were isolated from 20-22 day old rat testis and cultured on glass coverslips. These cells were grown to confluence with 10% fetal calf serum (FCS) in medium until day 4 and then subcultured to have secondary monocultures maintained with or without serum. By means of immunofluorescence and cytochemistry using avidin-biotin peroxidase complex it was observed that peritubular myoid cells were positive for FN and most of the FN was localized in the perinuclear region. Subcultured peritubular myoid cells maintained for 4 days in medium containing FCS developed an extensive interconnecting FN matrix. In the presence of 0.5 mM cAMP in culture, FN became localized along the filamentous process of peritubular myoid cells and more prominently in the areas of triangulated multi-cell aggregates as well as on the surface of the contracted small spherical cells. The addition of cAMP in the presence of FCS, also caused a noticeable change in the staining pattern; FN was detected along the filamentous process developing into a complex network of cells encased in an extensive matrix. It would appear that the translocation of FN in the cytoplasmic extensions of peritubular myoid cells may be a direct consequence of morphological changes associated with metabolic regulation of cAMP. This may also be related to the puberty associated development of in vivo changes in the ECM produced by peritubular myoid cells.

Accurately defining the reference condition for summary biotic metrics: A comparison of four approaches

Protocols for bioassessment often relate changes in summary metrics that describe aspects of biotic assemblage structure and function to environmental stress. Biotic assessment using multimetric indices now forms the basis for setting regulatory standards for stream quality and a range of other goals related to water resource management in the USA and elsewhere. Biotic metrics are typically interpreted with reference to the expected natural state to evaluate whether a site is degraded. It is critical that natural variation in biotic metrics along environmental gradients is adequately accounted for, in order to quantify human disturbance-induced change. A common approach used in the IBI is to examine scatter plots of variation in a given metric along a single stream size surrogate and a fit a line (drawn by eye) to form the upper bound, and hence define the maximum likely value of a given metric in a site of a given environmental characteristic (termed the 'maximum species richness line' - MSRL). In this paper we examine whether the use of a single environmental descriptor and the MSRL is appropriate for defining the reference condition for a biotic metric (fish species richness) and for detecting human disturbance gradients in rivers of south-eastern Queensland, Australia. We compare the accuracy and precision of the MSRL approach based on single environmental predictors, with three regression-based prediction methods (Simple Linear Regression, Generalised Linear Modelling and Regression Tree modelling) that use (either singly or in combination) a set of landscape and local scale environmental variables as predictors of species richness. We compared the frequency of classification errors from each method against set biocriteria and contrast the ability of each method to accurately reflect human disturbance gradients at a large set of test sites. The results of this study suggest that the MSRL based upon variation in a single environmental descriptor could not accurately predict species richness at minimally disturbed sites when compared with SLR's based on equivalent environmental variables. Regression-based modelling incorporating multiple environmental variables as predictors more accurately explained natural variation in species richness than did simple models using single environmental predictors. Prediction error arising from the MSRL was substantially higher than for the regression methods and led to an increased frequency of Type I errors (incorrectly classing a site as disturbed). We suggest that problems with the MSRL arise from the inherent scoring procedure used and that it is limited to predicting variation in the dependent variable along a single environmental gradient.

A comparison of spatially explicit landscape representation methods and their relationship to stream condition

1. Biodiversity, water quality and ecosystem processes in streams are known to be influenced by the terrestrial landscape over a range of spatial and temporal scales. Lumped attributes (i.e. per cent land use) are often used to characterise the condition of the catchment; however, they are not spatially explicit and do not account for the disproportionate influence of land located near the stream or connected by overland flow. 2. We compared seven landscape representation metrics to determine whether accounting for the spatial proximity and hydrological effects of land use can be used to account for additional variability in indicators of stream ecosystem health. The landscape metrics included the following: a lumped metric, four inverse-distance-weighted (IDW) metrics based on distance to the stream or survey site and two modified IDW metrics that also accounted for the level of hydrologic activity (HA-IDW). Ecosystem health data were obtained from the Ecological Health Monitoring Programme in Southeast Queensland, Australia and included measures of fish, invertebrates, physicochemistry and nutrients collected during two seasons over 4 years. Linear models were fitted to the stream indicators and landscape metrics, by season, and compared using an information-theoretic approach. 3. Although no single metric was most suitable for modelling all stream indicators, lumped metrics rarely performed as well as other metric types. Metrics based on proximity to the stream (IDW and HA-IDW) were more suitable for modelling fish indicators, while the HA-IDW metric based on proximity to the survey site generally outperformed others for invertebrates, irrespective of season. There was consistent support for metrics based on proximity to the survey site (IDW or HA-IDW) for all physicochemical indicators during the dry season, while a HA-IDW metric based on proximity to the stream was suitable for five of the six physicochemical indicators in the post-wet season. Only one nutrient indicator was tested and results showed that catchment area had a significant effect on the relationship between land use metrics and algal stable isotope ratios in both seasons. 4. Spatially explicit methods of landscape representation can clearly improve the predictive ability of many empirical models currently used to study the relationship between landscape, habitat and stream condition. A comparison of different metrics may provide clues about causal pathways and mechanistic processes behind correlative relationships and could be used to target restoration efforts strategically.

Cultural impact on e-service use in Saudi Arabia : the need for service oriented culture

This paper reports the results of a mixed method approach to answer: To what extent do cultural values impact on e-service use in Saudi Arabia, and if so how? This paper will firstly, introduce the importance of culture and define the aspects of Saudi culture with focus on our scope: the need for Service Oriented Culture. It will then, briefly, describe the method used and present the qualitative and quantitative findings related to the need for Service Oriented Culture. This research aims to cover a gap in the literature by investigating to what extent the presence of Service Oriented Culture, as one of Saudi Arabia’s cultural values, impacts on e-service use in Saudi Arabia. Surprisingly, the tested hypothesis was rejected: the presence of Service Oriented Culture is not a positive predictor of Intention to Use e-services in Saudi Arabia. It is evidenced that consideration of the impact of the cultural values will mainly contribute to the enhancement of ICTs implementation and use.

Safety culture and the australian heavy vehicle industry : a concept in chaos : an industry in need

This research proposed a new framework for safety culture and examined the influence that culture has on safety in the heavy vehicle industry. The results gave evidence for an industry wide culture, allowing future safety interventions to be designed in a culturally-relevant manner. Designing culturally-relevant interventions may maximise their effectiveness and reduce the levels of resistance to safety that have been evident in past years.

Condition monitoring and fault diagnosis of diesel engines using instantaneous angular speed analysis

Continuous monitoring of diesel engine performance is critical for early detection of fault developments in an engine before they materialize into a functional failure. Instantaneous crank angular speed (IAS) analysis is one of a few nonintrusive condition monitoring techniques that can be utilized for such a task. Furthermore, the technique is more suitable for mass industry deployments than other non-intrusive methods such as vibration and acoustic emission techniques due to the low instrumentation cost, smaller data size and robust signal clarity since IAS is not affected by the engine operation noise and noise from the surrounding environment. A combination of IAS and order analysis was employed in this experimental study and the major order component of the IAS spectrum was used for engine loading estimation and fault diagnosis of a four-stroke four-cylinder diesel engine. It was shown that IAS analysis can provide useful information about engine speed variation caused by changing piston momentum and crankshaft acceleration during the engine combustion process. It was also found that the major order component of the IAS spectra directly associated with the engine firing frequency (at twice the mean shaft rotating speed) can be utilized to estimate engine loading condition regardless of whether the engine is operating at healthy condition or with faults. The amplitude of this order component follows a distinctive exponential curve as the loading condition changes. A mathematical relationship was then established in the paper to estimate the engine power output based on the amplitude of this order component of the IAS spectrum. It was further illustrated that IAS technique can be employed for the detection of a simulated exhaust valve fault in this study.

The alcohol culture of mature-aged women : similarities and differences across borders

Introduction: Relatively few attempts have been made to describe and understand women’s alcohol consumption beyond adolescence and young adulthood. In particular, there has been a lack of studies focusing on the alcohol culture that surrounds and guides mature-aged women’s drinking. As part of a larger cross-national comparison, the present study sought to address this gap by identifying the shared beliefs and values that impact on drinking outcomes among mature-aged women in Sweden and Australia. Method: The study was guided by an ethnographic methodology. To generate data, a series of semi-structured interview were conducted with 17 Australian (age = 45-57 years; M = 52.1, SD = 3.9) and 19 Swedish (age = 45-58 years; M = 52.2, SD = 4.8) women. All interviews were transcribed verbatim and thematically analysed. Results: With age, the focus of alcohol as a single purpose vehicle for intoxication had given way to a focus on the enjoyment and ritual of drinking itself; taste had become increasingly important and alcohol was strongly associated with pleasurable environments and experiences. The view of alcohol as a taste experience was particularly pronounced among the Swedish women, with alcohol (most commonly wine) often seen as inseparable from food. Among the Swedish women, this view of alcohol was also associated with a strong de-emphasis of the pharmacological effects of alcohol. In contrast, several Australian women understood and used alcohol as relief for anxiety and stress. Moderate drinking was linked to the social construction of both age and gender in the two samples, while heavy or abusive alcohol consumption was associated with strong proscriptive norms and stigma. Conclusions: Australian and Swedish women share a number of beliefs and values around alcohol, however, these findings also show unique country-level differences. Implications for drinking outcomes are discussed.

Serial explant culture provides novel insights into the potential location and phenotype of corneal endothelial progenitor cells

The routine cultivation of human corneal endothelial cells, with the view to treating patients with endothelial dysfunction, remains a challenging task. While progress in this field has been buoyed by the proposed existence of progenitor cells for the corneal endothelium at the corneal limbus, strategies for exploiting this concept remain unclear. In the course of evaluating methods for growing corneal endothelial cells, we have noted a case where remarkable growth was achieved using a serial explant culture technique. Over the course of 7 months, a single explant of corneal endothelium, acquired from cadaveric human tissue, was sequentially seeded into 7 culture plates and on each occasion produced a confluent cell monolayer. Sample cultures were confirmed as endothelial in origin by positive staining for glypican-4. On each occasion, small cells, closest to the tissue explant, developed into a highly compact layer with an almost homogenous structure. This layer was resistant to removal with trypsin and produced continuous cell outgrowth during multiple culture periods. The small cells gave rise to larger cells with phase-bright cell boundaries and prominent immunostaining for both nestin and telomerase. Nestin and telomerase were also strongly expressed in small cells immediately adjacent to the wound site, following transfer of the explant to another culture plate. These findings are consistent with the theory that progenitor cells for the corneal endothelium reside within the limbus and provide new insights into expected expression patterns for nestin and telomerase within the differentiation pathway.

A test rig for the condition-based maintenance application on the traction chain of very high speed trains

The preventive maintenance of traction equipment for Very High Speed Trains (VHST) nowadays is becoming very expensive owing to the high complexity and quality of these components that require high reliability. An efficient maintenance approach like the Condition-Based Maintenance (CBM) should be implemented to reduce the costs. For this purpose, an experimental full-scale test rig for the CBM of VHST traction equipment has been designed to investigate in detail failures in the main mechanical components of system, i.e. motor, bearings and gearbox. The paper describes the main characteristics of this unique test rig, able to reproduce accurately the train operating conditions, including the relative movements of the motor, the gearbox and the wheel axle. Gearbox, bearing seats and motor are equipped by accelerometers, thermocouples, torque meter and other sensors in different positions. The testing results give important information about the most suitable sensor position and type to be installed for each component and show the effectiveness of the techniques used for the signal analysis in order to identify faults of the gearbox and motor bearings.

Doctoral research training : encouraging timely completion – case study reflections of one university on how it supports a vibrant research training culture

This is a case study of a young university striving to generate and sustain a vibrant Research Training culture. The university’s research training framework is informed by a belief in a project management approach to achieving successful research candidature. This has led to the definition and reporting of key milestones during candidature. In turn, these milestones have generated a range of training programs to support Higher Degree Research (HDR) students to meet these milestones in a timely fashion. Each milestone focuses on a specific set of skills blended with supporting the development of different parts of the doctoral thesis. Data on student progress and completion has provided evidence in highlighting the role that the milestones and training are playing in supporting timely completion. A university-wide reporting cycle generated data on the range of workshops and training provided to Higher Degree Research students and supervisors. The report provided details of thesis topic and format, as well as participation in research training events and participant evaluation of those events. Analysis of the data led to recommendations and comments on the strengths and weaknesses of the current research training program. Discussion considered strategies and drivers for enhancements into the future. In particular, the paper reflects on the significant potential role of centrally curated knowledge systems to support HDR student and supervisor access, and engagement and success. The research training program was developed using blended learning as a model. It covered face-to-face workshops as well as online modules. These were supplemented by web portals that offered a range of services to inform and educate students and supervisors and included opportunities for students to interact with each other. Topics ranged from the research life cycle, writing and publication, ethics, managing research data, managing copyright, and project management to use of software and the University’s Code of Conduct for Research. The challenges discussed included: How to reach off campus students and those studying in external modes? How best to promote events to potential participants? How long and what format is best for face-to-face sessions? What online resources best supplement face-to-face offerings? Is there a place for peer-based learning and what form should this take? These questions are raised by a relatively young university seeking to build and sustain a vibrant research culture. The rapid growth in enrolments in recent years has challenged previous one-to-one models of support. This review of research training is timely in seeking strategies to address changing research training support capacity and student needs. Part of the discussion will focus on supervisory training, noting that good supervision is the one remaining place where one-to-one support is provided. Ensuring that supervisors are appropriately equipped to address student expectations is considered in the context of the research training provisions. The paper concludes with reflection on the challenges faced, and recommended ways forward as the number of research students grows into the future.