Investigation of molecular mechanisms regulating biomineralization of pearl oyster Pinctada maxima

Biomineralization is a process encompassing all mineral containing tissues produced within an organism. The most dynamic example of this process is the formation of the mollusk shell, comprising a variety of crystal phases and microstructures. The organic component incorporated within the shell is said to dictate this remarkable architecture. Subsequently, for the past decade considerable research have been undertaken to identify and characterize the protein components involved in biomineralization. Despite these efforts the general understanding of the process remains ambiguous. This study employs a novel molecular approach to further the elucidation of the shell biomineralization. A microarray platform has been custom generated (PmaxArray 1.0) from the pearl oyster Pinctada maxima. PmaxArray 1.0 consists of 4992 expressed sequence tags (ESTs) originating from the mantle, an organ involved in shell formation. This microarray has been used as the primary tool for three separate investigations in an effort to associate transcriptional gene expression from P. maxima to the process of shell biomineralization. The first investigation analyzes the spatial expression of ESTs throughout the mantle organ. The mantle was dissected into five discrete regions and each analyzed for gene expression with PmaxArray 1.0. Over 2000 ESTs were differentially expressed among the tissue sections, identifying five major expression regions. Three of these regions have been proposed to have shell formation functions belonging to nacre, prismatic calcite and periostracum. The spatial gene expression map was confirmed by in situ hybridization, localizing a subset of ESTs from each expression region to the same mantle area. Comparative sequence analysis of ESTs expressed in the proposed shell formation regions with the BLAST tool, revealed a number of the transcripts were novel while others showed significant sequence similarities to previously characterized shell formation genes. The second investigation correlates temporal EST expression during P. maxima larval ontogeny with transitions in shell mineralization during the same period. A timeline documenting the morphologicat microstructural and mineralogical shell characteristics of P. maxima throughout larval ontogeny has been established. Three different shell types were noted based on the physical characters and termed, prodissoconch I, prodissoconch 11 and dissoconch. PmaxArray 1.0 analyzed ESTs expression of animals throughout the larval development of P. maxima, noting up-regulation of 359 ESTs in association with the shell transitions from prodissoconch 1 to prodissoconch 11 to dissoconch. Comparative sequence analysis of these ESTs indicates a number of the transcripts are novel as well as showing significant sequence similarities between ESTs and known shell matrix associated genes and proteins. These ESTs are discussed in relation to the shell characters associated with their temporal expression. The third investigation uses PmaxArray 1.0 to analyze gene expression in the mantle tissue of P. maxima specimens exposed to sub-lethal concentrations of a shell-deforming toxin, tributyltin (TBT). The shell specific effects of TBT are used in this investigation to interpret differential expression of ESTs with respect to shell formation functions. A lethal and sublethal TBT concentration range was established for P. maxima, noting a concentration of 50 ng L- 1 TBT as sub-lethal over a 21 day period. Mantle tissue from P. maxima animals treated with 50 ng L- 1 TBT was assessed for differential EST expression with untreated control animals. A total of 102 ESTs were identified as differentially expressed in association with TBT exposure, comparative sequence identities included an up-regulation of immunity and detoxification related genes and down-regulation of several shell matrix genes. A number of transcripts encoding novel peptides were additionally identified. The potential actions of these genes are discussed with reference to TBT toxicity and shell biomineralization. This thesis has used a microarray platform to analyze gene expression in spatial, temporal and toxicity investigations, revealing the involvement of numerous gene transcripts in specific shell formation functions. Investigation of thousands of transcripts simultaneously has provided a holistic interpretation of the organic components regulating shell biomineralization.

Plasma ATP concentration and venous oxygen content in the forearm during dynamic handgrip exercise

Background: It has been proposed that adenosine triphosphate (ATP) released from red blood cells (RBCs) may contribute to the tight coupling between blood flow and oxygen demand in contracting skeletal muscle. To determine whether ATP may contribute to the vasodilatory response to exercise in the forearm, we measured arterialised and venous plasma ATP concentration and venous oxygen content in 10 healthy young males at rest, and at 30 and 180 seconds during dynamic handgrip exercise at 45% of maximum voluntary contraction (MVC). Results: Venous plasma ATP concentration was elevated above rest after 30 seconds of exercise (P < 0.05), and remained at this higher level 180 seconds into exercise (P < 0.05 versus rest). The increase in ATP was mirrored by a decrease in venous oxygen content. While there was no significant relationship between ATP concentration and venous oxygen content at 30 seconds of exercise, they were moderately and inversely correlated at 180 seconds of exercise (r = -0.651, P = 0.021). Arterial ATP concentration remained unchanged throughout exercise, resulting in an increase in the venous-arterial ATP difference. Conclusions: Collectively these results indicate that ATP in the plasma originated from the muscle microcirculation, and are consistent with the notion that deoxygenation of the blood perfusing the muscle acts as a stimulus for ATP release. That ATP concentration was elevated just 30 seconds after the onset of exercise also suggests that ATP may be a contributing factor to the blood flow response in the transition from rest to steady state exercise.

An examination of communication across cultures in news media and at informal/personal levels : with concentration on relations among two South East Asian countries and Australia and those two countries and Germany

In the age of globalisation dominated by mass communication, the flow of information contributes to a big extent to the worldviews of its "global citizens". From this point of view the mass media can be seen as one of the most salient sources of cross-cultural communication. This study investigates mass communication across cultures, focusing on South East Asia (Malaysia and Singapore), Australia and Germany. The centre of attention is the Western media coverage of South East Asia and vice versa. In this context a content analysis of newspapers of the three regions has been conducted. In addition, working practices and conditions of Western foreign correspondents in South East Asia have been examined. Apart from the investigation of inter-cultural media coverage, another focus of attention will be the examination of two levels of communication: The business level, concentrating on issues like e.g. the Asian business etiquette; and the private level, looking into the transition to a different culture from the perspective of Australian and German expatriates. Apart from investigating mass communication across cultures and to provide a written analysis of the findings, a series of radio documentaries in English and in German has been produced. They cover the following issues: Foreign correspondents in South East Asia, the expatriate-lifestyle of Australians and Germans in South East Asia, business etiquette in Asia, student exchange Germany-Asia, image and prejudices East-West and Tourism.

Corona ions from overhead transmission voltage powerlines : effect on DC e-field and ambient particle concentration levels

Along with their essential role in electricity transmission and distribution, some powerlines also generate large concentrations of corona ions. This study aimed at comprehensive investigation of corona ions, vertical dc e-field, ambient aerosol particle charge and particle number concentration levels in the proximity of some high/sub-transmission voltage powerlines. The influence of meteorology on the instantaneous value of these parameters, and the possible existence of links or associations between the parameters measured were also statistically investigated. The presence of positive and negative polarities of corona ions was associated with variation in the mean vertical dc e-field, ambient ion and particle charge concentration level. Though these variations increased with wind speed, their values also decreased with distance from the powerlines. Predominately positive polarities of ions were recorded up to a distance of 150 m (with the maximum values recorded 50 m downwind of the powerlines). At 200 m from the source, negative ions predominated. Particle number concentration levels however remained relatively constant (103 particle cm-3) irrespective of the sampling site and distance from the powerlines. Meteorological factors of temperature, humidity and wind direction showed no influence on the electrical parameters measured. The study also discovered that e-field measurements were not necessarily a true representation of the ground-level ambient ion/particle charge concentrations.

Young children’s active citizenship : storytelling, stories, and social actions

This thesis inquires into possibilities for young children‘s active citizenship as provoked through a practice of social justice storytelling with one Preparatory1 class of children aged five to six years. The inquiry was practitioner-research, through a living educational theory approach cultivating an interrelational view of existing with others in evolving processes of creation. Ideas of young children‘s active citizenship were provoked and explored through storytelling, by a storytelling teacher-researcher, a Prep class of children and their teacher. The three major foci of the study were practice, narrative and action. A series of storytelling workshops with a Prep class was the practice that was investigated. Each workshop began with a story that made issues of social justice visible, followed by critical discussion of the story, and small group activities to further explore the story. The focus on narrative was based on the idea of story as a way knowing. Stories were used to explore social justice issues with young children. Metanarratives of children and citizenship were seen to influence possibilities for young children‘s active citizenship. Stories were purposefully shared to provoke and promote young children‘s active citizenship through social actions. It was these actions that were the third focus of the study. Through action research, a social justice storytelling practice and the children‘s responses to the stories were reflected on both in action and after. These reflections informed and shaped storytelling practice. Learning in a practice of social justice storytelling is explained through living theories of social justice storytelling as pedagogy. Data of the children‘s participation in the study were analysed to identify influences and possibilities for young children‘s active citizenship creating a living theory of possibilities for young children‘s active citizenship.

Indoor and outdoor nitrogen dioxide concentration in residential houses in Australia

As part of a larger indoor environmental study, residential indoor and outdoor levels of nitrogen dioxide (NO2) were measured for 14 houses in a suburb of Brisbane, Queensland, Australia. Passive samplers were used for 48-h sampling periods during the winter of 1999. The average indoor and outdoor NO2 levels were 13.8 ± 6.3 and 16.7 ± 4.2 ppb, respectively. The indoor/outdoor NO2 concentration ratio ranged from 0.4 to 2.3, with a median value of 0.82. The results of statistic analyses indicated that there was no significant correlation between indoor and outdoor NO2 concentrations, or between indoor and fixed site NO2 monitoring station concentrations. However, there was a significant correlation between outdoor and fixed site NO2 monitoring station concentrations. There was also a significant correlation between indoor NO2 concentration and indoor submicrometre (0.007–0.808 μm) aerosol particle number concentrations. The results in this study indicated indoor NO2 levels are significantly affected by indoor NO2 sources, such as a gas stove and cigarette smoking. It implies that the outdoor or fixed site monitoring concentration alone is a poor predictor of indoor NO2 concentration.

Cluster analysis of movement patterns in multiarticular actions: A tutorial

The present paper proposes a technical analysis method for extracting information about movement patterning in studies of motor control, based on a cluster analysis of movement kinematics. In a tutorial fashion, data from three different experiments are presented to exemplify and validate the technical method. When applied to three different basketball-shooting techniques, the method clearly distinguished between the different patterns. When applied to a cyclical wrist supination-pronation task, the cluster analysis provided the same results as an analysis using the conventional discrete relative phase measure. Finally, when analyzing throwing performance constrained by distance to target, the method grouped movement patterns together according to throwing distance. In conclusion, the proposed technical method provides a valuable tool to improve understanding of coordination and control in different movement models, including multiarticular actions.

Experimental study of the concentration field of discharge from a boat propeller

Two-stroke outboard boat engines using total loss lubrication deposit a significant proportion of their lubricant and fuel directly into the water. The purpose of this work is to document the velocity and concentration field characteristics of a submerged swirling water jet emanating from a propeller in order to provide information on its fundamental characteristics. The properties of the jet were examined far enough downstream to be relevant to the eventual modelling of the mixing problem. Measurements of the velocity and concentration field were performed in a turbulent jet generated by a model boat propeller (0.02 m diameter) operating at 1500 rpm and 3000 rpm in a weak co-flow of 0.04 m/s. The measurements were carried out in the Zone of Established Flow up to 50 propeller diameters downstream of the propeller, which was placed in a glass-walled flume 0.4 m wide with a free surface depth of 0.15 m. The jet and scalar plume development were compared to that of a classical free round jet. Further, results pertaining to radial distribution, self similarity, standard deviation growth, maximum value decay and integral fluxes of velocity and concentration were presented and fitted with empirical correlations. Furthermore, propeller induced mixing and pollutant source concentration from a two-stroke engine were estimated.

The spermostatic and microbicidal actions of quinones and malemides : towards a dual-purpose contraceptive agent

There is an urgent need to develop safe, effective, dual-purpose contraceptive agents that combine the prevention of pregnancy with protection against sexually transmitted diseases. Here we report the identification of a group of compounds that on contact with human spermatozoa induce a state of “spermostasis,” characterized by the extremely rapid inhibition of sperm movement without compromising cell viability. These spermostatic agents were more active and significantly less toxic than the reagent in current clinical use, nonoxynol 9, giving therapeutic indices (ratio of spermostatic to cytotoxic activity) that were orders of magnitude greater than this traditional spermicide. Although certain compounds could trigger reactive oxygen species generation by spermatozoa, this activity was not correlated with spermostasis. Rather, the latter was associated with alkylation of two major sperm tail proteins that were identified as A Kinase-Anchoring Proteins (AKAP3 and AKAP4) by mass spectrometry. As a consequence of disrupted AKAP function, the abilities of cAMP to drive protein kinase A-dependent activities in the sperm tail, such as the activation of SRC and the consequent stimulation of tyrosine phosphorylation, were suppressed. Furthermore, analysis of microbicidal activity using Chlamydia muridarum revealed powerful inhibitory effects at the same low micromolar doses that suppressed sperm movement. In this case, the microbicidal action was associated with alkylation of Major Outer Membrane Protein (MOMP), a major chlamydial membrane protein. Taken together, these results have identified for the first time a novel set of cellular targets and chemical principles capable of providing simultaneous defense against both fertility and the spread of sexually transmitted disease.