Genetic resources in wild and cultured stocks of the Asian mud crab, Scylla paramamosain

The mud crab (Scylla spp.) aquaculture industry has expanded rapidly in recent years in many countries in the Indo - West Pacific (IWP) region as an alternative to marine shrimp culture because of significant disease outbreaks and associated failures of many shrimp culture industries in the region. Currently, practices used to produce and manage breeding crabs in hatcheries may compromise levels of genetic diversity, ultimately compromising growth rates, disease resistance and stock productivity. Therefore, to avoid “genetic pollution” and its harmful effects and to promote further development of mud crab aquaculture and fisheries in a sustainable way, a greater understanding of the genetic attributes of wild and cultured mud crab stocks is required. Application of these results can provide benefits for managing wild and cultured Asian mud crab populations for multiple purposes including for commercial production, recreation and conservation and to increase profitability and sustainability of newly emerging crab culture industries. Phylogeographic patterns and the genetic structure of Asian mud crab populations across the IWP were assessed to determine if they were concordant with those of other widespread taxa possessing pelagic larvae of relatively long duration. A 597 bp fragment of the mitochondrial DNA COI gene was amplified and screened for variation in a total of 297 individuals of S. paramamosain from six sampling sites across the species’ natural geographical distribution in the IWP and 36 unique haplotypes were identified. Haplotype diversities per site ranged from 0.516 to 0.879. Nucleotide diversity estimates among haplotypes were 0.11% – 0.48%. Maximum divergence observed among S. paramamosain samples was 1.533% and samples formed essentially a single monophyletic group as no obvious clades were related to geographical location of sites. A weak positive relationship was observed however, between genetic distance and geographical distance among sites. Microsatellite markers were then used to assess contemporary gene flow and population structure in Asian mud crab populations sampled across their natural distribution in the IWP. Eight microsatellite loci were screened in sampled S. paramamosain populations and all showed high allelic diversity at all loci in sampled populations. In total, 344 individuals were analysed, and 304 microsatellite alleles were found across the 8 loci. The mean number of alleles per locus at each site ranged from 20.75 to 28.25. Mean allelic richness per site varied from 17.2 to 18.9. All sites showed high levels of heterozygosity as average expected heterozygosities for all loci ranged from 0.917 – 0.953 while mean observed heterozygosity ranged from 0.916 – 0.959. Allele diversities were similar at all sites and across all loci. The results did not show any evidence for major differences in allele frequencies among sites and patterns of allele frequencies were very similar in all populations across all loci. Estimates of population differentiation (FST) were relatively low and most probably largely reflect intra – individual variation for very highly variable loci. Results from nDNA analysis showed evidence for only very limited population genetic structure among sampled S. paramamosain, and a positive and significant association for genetic and geographical distance among sample sites. Microsatellite markers were then employed to determine if adequate levels of genetic diversity has been captured in crab hatcheries for the breeding cycle. The results showed that all microsatellite loci were polymorphic in hatchery samples. Culture populations were in general, highly genetically depauperate, compared with comparable wild populations, with only 3 to 8 alleles recorded for the same loci set per population. In contrast, very high numbers of alleles per locus were found in reference wild S. paramamosain populations, which ranged from 18 to 46 alleles per locus per population. In general, this translates into a 3 to 10 fold decline in mean allelic richness per locus in all culture stocks compared with wild reference counterparts. Furthermore, most loci in all cultured S. paramamosain samples showed departures from HWE equilibrium. Allele frequencies were very different in culture samples from that present in comparable wild reference samples and this in particular, was reflected in a large decline in allele diversity per locus. The pattern observed was best explained by significant impacts of breeding practices employed in hatcheries rather than natural differentiation among wild populations used as the source of brood stock. Recognition of current problems and management strategies for the species both for the medium and long-term development of the new culture industry are discussed. The priority research to be undertaken over the medium term for S. paramamosain should be to close the life cycle fully to allow individuals to be bred on demand and their offspring equalised to control broodstock reproductive contributions. Establishing a broodstock register and pedigree mating system will be required before any selection program is implemented. This will ensure that sufficient genetic variation will be available to allow genetic gains to be sustainably achieved in a future stock improvement program. A fundamental starting point to improve hatchery practices will be to encourage farmers and hatchery managers to spawn more females in their hatcheries as it will increase background genetic diversity in culture stocks. Combining crablet cohorts from multiple hatcheries into a single cohort for supply to farmers or rotation of breeding females regularly in hatcheries will help to address immediate genetic diversity problems in culture stocks. Application of these results can provide benefits for managing wild and cultured Asian mud crab populations more efficiently. Over the long-term, application of data on genetic diversity in wild and cultured stocks of Asian mud crab will contribute to development of sustainable and productive culture industries in Vietnam and other countries in the IWP and can contribute towards conservation of wild genetic resources.

Increased Langerhan cell density and corneal nerve damage in diabetic patients : role of immune mechanisms in human diabetic neuropathy

Aim/hypothesis Immune mechanisms have been proposed to play a role in the development of diabetic neuropathy. We employed in vivo corneal confocal microscopy (CCM) to quantify the presence and density of Langerhans cells (LCs) in relation to the extent of corneal nerve damage in Bowman's layer of the cornea in diabetic patients. Methods 128 diabetic patients aged 58±1 yrs with a differing severity of neuropathy based on Neuropathy Deficit Score (NDS—4.7±0.28) and 26 control subjects aged 53±3 yrs were examined. Subjects underwent a full neurological evaluation, evaluation of corneal sensation with non-contact corneal aesthesiometry (NCCA) and corneal nerve morphology using corneal confocal microscopy (CCM). Results The proportion of individuals with LCs was significantly increased in diabetic patients (73.8%) compared to control subjects (46.1%), P=0.001. Furthermore, LC density (no/mm2) was significantly increased in diabetic patients (17.73±1.45) compared to control subjects (6.94±1.58), P=0.001 and there was a significant correlation with age (r=0.162, P=0.047) and severity of neuropathy (r=−0.202, P=0.02). There was a progressive decrease in corneal sensation with increasing severity of neuropathy assessed using NDS in the diabetic patients (r=0.414, P=0.000). Corneal nerve fibre density (P<0.001), branch density (P<0.001) and length (P<0.001) were significantly decreased whilst tortuosity (P<0.01) was increased in diabetic patients with increasing severity of diabetic neuropathy. Conclusion Utilising in vivo corneal confocal microscopy we have demonstrated increased LCs in diabetic patients particularly in the earlier phases of corneal nerve damage suggestive of an immune mediated contribution to corneal nerve damage in diabetes.

Conceptualising convenience : transportation practices and perceptions of inner-urban high density residents in Brisbane, Australia

High-density living in inner-urban areas has been promoted to encourage the use of more sustainable modes of travel to reduce greenhouse gas emissions. However, previous research presents mixed results on the relationship between living in proximity to transport systems and reduced car-dependency. This research examines inner-city residents’ transportation practices and perceptions, via 24 qualitative interviews with residents from high-density dwellings in inner-city Brisbane, Australia. Whilst participants consider public transport accessible and convenient, car use continues to be relied on for many journeys. Transportation choices are justified through complex definitions of convenience containing both utilitarian and psycho-social elements,with three key themes identified: time-efficiency, single versus multi-modal trips, and distance to and purpose of journey, as well as attitudinal, affective and symbolic elements related to transport mode use. Understanding conceptions of transport convenience held by different segments of the transport users market,alongside other factors strongly implicated in travel mode choice, can ensure targeted improvements in sustainable transport service levels and infrastructure as well as information service provision and behavioural change campaigns.

Project report. A subtropical urban community: Investigating medium to high density residential typologies by Design Charrette

The Centre for Subtropical Design at QUT, in partnership with the Queensland Government and Brisbane City Council, conducts research focused on 'best practice' outcomes for higher density urban living environments in the subtropics through the study of typical urban residential typologies, and urban design. The aim of the research is to inform and illustrate best practice subtropical design principles to policy makers and development industry professionals to stimulate climate-responsive outcomes. The Centre for Subtropical Design recently sought project-specific funding from the Queensland Department of Infrastructure and Planning (DIP) to investigate residential typologies for sustainable subtropical urban communities, based on transit orientated development principles and outcomes for areas around public transport nodes. A development site within the Fitzgibbon Urban Development Area, and close to a rail and bsu transport corridor, provided a case study location for this project. Four design-led multi-disciplinary creative teams participated in a Design Charrette and have produced concept drawings and propositions on a range of options, or prototypes. Analysis of selected prototypes has been undertaken to determine their environmental, economic and social performance. This Project Report discusses the scope of the project funded by DIP in terms of activities undertaken to date, and deliverables achieved. A subsequent Research Report will discuss the detailed findings of the analysis.

Research report. A subtropical urban community. Investigating medium to high density residential typologies. Findings and recommendations from the Design Charrette

In June 2009 the Centre for Subtropical Design at the Queensland University of Technology conducted a design charrette to research design concepts for liveable subtropical neighbourhoods characterised by higher-density, mixed-use, family orientated housing. Subsequent analysis of the proposed designs evaluated how well these typologies support economic, environmental and social sustainability. The study was led by Ms Rosemary Kennedy, Director of the Centre for Subtropical Design and QUT School of Design Adjunct Professor Peter Richards, Chair of the Centre for Subtropical Design Board and director of Deicke Richards Architects and Urban Designers.

A subtropical urban community. Investigating medium to high density residential typologies. Recommendations from the design charrette

In June 2009 the Centre for Subtropical Design at the Queensland University of Technology conducted a design charrette to research design concepts for liveable subtropical neighbourhoods characterised by higher-density, mixed-use, family orientated housing. Subsequent analysis of the proposed designs evaluated how well these typologies support economic, environmental and social sustainability. The study was led by Ms Rosemary Kennedy, Director of the Centre for Subtropical Design and QUT School of Design Adjunct Professor Peter Richards, Chair of the Centre for Subtropical Design Board and director of Deicke Richards Architects and Urban Designers.

Case study 1 : a subtropical urban community : investigating medium to high density residential typologies by design charrette

The public transport corridor bordering the study site runs NW to SE and is perceived as a source of noise and pollution. The key urban planning strategies adopted by this team were: • Acoustic separation from transport corridor noise source, • A regular grid pattern of urban blocks, and • A clear hierarchy of accessible open space throughout the development.

Report : Case study 2 : a subtropical urban community : investigating medium to high density residential typologies by design charrette

The case study site is physically disconnected from its surrounding community by the rail corridor and future bus lanes and is unlikely to be able to sustain its own commercial retail centre. As a result, it may also be socially disconnected from surrounding suburbs. However, it does offer proximity and access to an extensive „natural‟ area, and this is seen as key opportunity for the proposed development to develop a strong relationship with surrounding suburbs...

Decentralised Data Fusion with Parzen Density Estimates

Decentralised sensor networks typically consist of multiple processing nodes supporting one or more sensors. These nodes are interconnected via wireless communication. Practical applications of Decentralised Data Fusion have generally been restricted to using Gaussian based approaches such as the Kalman or Information Filter This paper proposes the use of Parzen window estimates as an alternate representation to perform Decentralised Data Fusion. It is required that the common information between two nodes be removed from any received estimates before local data fusion may occur Otherwise, estimates may become overconfident due to data incest. A closed form approximation to the division of two estimates is described to enable conservative assimilation of incoming information to a node in a decentralised data fusion network. A simple example of tracking a moving particle with Parzen density estimates is shown to demonstrate how this algorithm allows conservative assimilation of network information.

A subtropical urban community : medium to high density residential typologies - case study 4

On the case study site, using these strategies, the site density achieved was approximately 180 dwellings per hectare. According to ASK consulting engineers‟ acoustic report (in Ecolateral‟s report) the design gives solid consideration to the environmental noise issues associated with the site. The subject structure not only provides significant shielding of transport corridor noise to the suburb, it also minimises the potential for adverse impact on residential amenity within the building itself...

Porphyromonas gingivalis lipopolysaccharide alters atherosclerotic-related gene expression in oxidized low-density-lipoprotein-induced macrophages and foam cells

The molecular mechanism between atherosclerosis formation and periodontal pathogens is not clear although positive correlation between periodontal infections and cardiovascular diseases has been reported. Objective: To determine if atherosclerosis related genes were affected in foam cells during and after its formation by P. gingivalis lipopolysaccharide (LPS) stimulation. Methods: Macrophages from human THP-1 monocytes were treated with oxidized low density lipoprotein (oxLDL) to induce the formation of foam cells. P. gingivalis LPS was added to cultures of either oxLDL-induced macrophages or foam cells. The expression of atherosclerosis related genes was assayed by quantitative real time PCR and the protein production of granulocyte-macrophage colony-stimulating factor（GM-CSF）, monocyte chemotactic protein-1 (MCP-1), IL-1β, IL-10 and IL-12 was determined by ELISA. Nuclear translocation of NF-κB P65 was detected by immunocytochemistry and western blot was used to evaluate IKB-α degradation to confirm the NF-κB pathway activation. Results: P. gingivalis LPS stimulated atherosclerosis related gene expression in foam cells and increased oxLDL induced expression of chemokines, adhesion molecules, growth factors, apoptotic genes, and nuclear receptors in macrophages. Transcription of the pro-inflammatory cytokines IL-1β and IL-12 was elevated in response to LPS in both macrophages and foam cells, whereas the anti-inflammatory cytokine IL-10 was not affected. Increased NF-κB pathway activation was also observed in LPS and oxLDL co-stimulated macrophages. Conclusion: P. gingivalis LPS appears to be an important factor in the development of atherosclerosis by stimulation of atherosclerosis related gene expression in both macrophages and foam cells via activation of the NF-κB pathway.

Cell density alters matrix accumulation in two distinct fractions and the mechanical integrity of alginate-chondrocyte constructs

Chondrocyte density in articular cartilage is known to change with the development and growth of the tissue and may play an important role in the formation of a functional extracellular matrix (ECM). The objective of this study was to determine how initial chondrocyte density in an alginate hydrogel affects the matrix composition, its distribution between the cell-associated (CM) and further removed matrix (FRM) fractions, and the tensile mechanical properties of the developing engineered cartilage. Alginate constructs containing primary bovine chondrocytes at densities of 0, 4, 16, and 64 million cells/ml were fabricated and cultured for 1 or 2 weeks, at which time structural, biochemical, and mechanical properties were analyzed. Both matrix content and distribution varied with the initial cell density. Increasing cell density resulted in an increasing content of collagen and sulfated-glycosaminoglycan (GAG) and an increasing proportion of these molecules localized in the CM. While the equilibrium tensile modulus of cell-free alginate did not change with time in culture, the constructs with highest cell density were 116% stiffer than cell-free controls after 2 weeks of culture. The equilibrium tensile modulus was positively correlated with total collagen (r2 = 0.47, p < 0.001) and GAG content (r2 = 0.68, p < 0.001), and these relationships were enhanced when analyzing only those matrix molecules in the CM fraction (r2 = 0.60 and 0.72 for collagen and GAG, respectively, each p < 0.001). Overall, the results of this study indicate that initial cell density has a considerable effect on the developing composition, structure, and function of alginate–chondrocyte constructs.