Packed bed bioreactor for the isolation and expansion of placental-derived Mesenchymal Stromal Cells

Large numbers of Mesenchymal stem/stromal cells (MSCs) are required for clinical relevant doses to treat a number of diseases. To economically manufacture these MSCs, an automated bioreactor system will be required. Herein we describe the development of a scalable closed-system, packed bed bioreactor suitable for large-scale MSCs expansion. The packed bed was formed from fused polystyrene pellets that were air plasma treated to endow them with a surface chemistry similar to traditional tissue culture plastic. The packed bed was encased within a gas permeable shell to decouple the medium nutrient supply and gas exchange. This enabled a significant reduction in medium flow rates, thus reducing shear and even facilitating single pass medium exchange. The system was optimised in a small-scale bioreactor format (160 cm2) with murine-derived green fluorescent protein-expressing MSCs, and then scaled-up to a 2800 cm2 format. We demonstrated that placental derived MSCs could be isolated directly within the bioreactor and subsequently expanded. Our results demonstrate that the closed system large-scale packed bed bioreactor is an effective and scalable tool for large-scale isolation and expansion of MSCs.

Anomeric effect in sulfur-containing systems: An Ab-initio study

The anomeric effect in S---C---S and O---C---S systems was studied by using closed-shell Hartree-Fock theory. A comparison of the STO-3G level with the 4–31G and 6–31G* levels was performed for the O---C---O system, and the STO-3G level found adequate for study of the anomeric effect. Optimization of bond lengths and angles was conducted at the STO-3G level and limited studies were made at the 4–31G level. The nature of the torsional potential curves is compared for the O---C---O, O---C---S, and S---C---S systems. The possible reasons for the decreased anomeric effect in sulfur systems are discussed.

Crystal and molecular structure of the quinoxaline antibiotic analog TANDEM (des-N-tetramethyltriostin A)

The crystal structure of TANDEM (des-N-tetramethyltriostin A), a synthetic analogue of the quinoxaline antibiotic triostin A, has been determined independently at -135 and 7 'C and refined to R values of 0.088 and 0.147, respectively. The molecule has approximate 2-fold symmetry, with the quinoxaline chromophores and the disulfide cross-bridge projecting from opposite sides of the peptide ring. The quinoxaline groups are nearly parallel to each other and separated by about 6.5 A. The peptide backbone resembles a distorted antiparallel 13 ribbon joined by intramolecular hydrogen bonds N-H(LVal)--O(L-Ala). At low temperatures, the TANDEM molecule is surrounded by a regular first- and second-order hydration sphere containing 14 independent water molecules. At room temperature, only the first-order hydration shell is maintained. Calculations of the interplanar separation of the quinoxaline groups as a function of their orientation with respect to the peptide ring support the viability of TANDEM to intercalate bifunctionally into DNA.

Nitrogen use for high productivity and sustainability in cashew.

Interest in cashew production in Australia has been stimulated by domestic and export market opportunities and suitability of large areas of tropical Australia. Economic models indicate that cashew production is profitable at 2.8 t ha-1 nut-in-shell (NIS). Balanced plant nutrition is essential to achieve economic yields in Australia, with nitrogen (N) of particular importance because of its capacity to modify growth, affect nut yield and cause environmental degradation through soil acidification and off-site contamination. The study on a commercial cashew plantation at Dimbulah, Australia, investigated the effect of N rate and timing on cashew growth, nut production, N leaching and soil chemical properties over five growth cycles (1995-1999). Nitrogen was applied during the main periods of vegetative (December-April) and reproductive (June-October) growth. Commercial NIS yields (up to 4.4 t ha-1 from individual trees) that exceeded the economic threshold of 2.8 t ha-1 were achieved. The yield response was mainly determined by canopy size as mean nut weight, panicle density and nuts per panicle were largely unaffected by N treatments. Nitrogen application confined to the main period of vegetative growth (December-April) produced a seasonal growth pattern that corresponded most consistently with highest NIS yield. This N timing also reduced late season flowering and undesirable post-November nut drop. Higher yields were not produced at N rates greater than 17 g m-2 of canopy surface area (equating to 210 kg N ha-1 for mature size trees). High yields were attained when N concentrations in Mveg leaves in May-June were about 2%, but this assessment occurs at a time when it is not feasible to correct N deficiency. The Mflor leaf of the preceding November, used in conjunction with the Mveg leaf, was proposed as a diagnostic tool to guide N rate decisions. Leaching of nitrate-N and acidification of the soil profile was recorded to 0.9 m. This is an environmental and sustainability hazard, and demonstrates that improved methods of N management are required.

Hole Pairing In Antiferromagnetically Correlated Layered Oxides

We have considered a two-band Hubbard model having interlaced Cu-3d(x2−y2) and O-2p(x, y) orbitals representing the CuO2 square planes. Simple CuO2 -cluster calculation suggests that the additional holes created by doping stay mainly on oxygen. Motion of an oxygen hole interlacing with the antiferromagnetically correlated background of copper spins, creates a string of high energy spin configuration of finite length giving mass renormalization. Another hole of opposite spin can now anneal this string tension providing a triangular pairing potential for large pair momentum. The latter implies unusual Bose condensation of the wake-bound compact Bose-like pairs on a non-zero momentum shell. Effect of disorder favouring condensation at the mobility edge is pointed out.

Raw data for project "Development of a DNA based aging technique for use in fisheries assessments" Fisheries and Development Corporation project 2007/033

The primary aim of this study was to determine the relationship between telomere length and age in a range of marine invertebrates including abalone (Haliotis spp) oysters (Saccostrea glomerata), spiny lobsters (Sagmariasus verreauxi formerly Jasus verreauxi and Jasus edwardsii) and school prawns (Metapenaeus macleayi). Additionally, this relationship was studied in a vertebrate organism using the freshwater fish Silver perch (Bidyanus bidyanus). Telomere length differences between tissues were also examined in some species such as Saccostrea glomerata, Sagmariasus verreauxi and Bidyanus bidyanus. In some cases cultured specimens of known age were used and this is quoted in the spreadsheets. For other wild-caught specimens where age was not known, size was used as a proxy for age. This may be a broad size class, or be determined by shell size or carapace length depending on the organism. Each spreadsheet contains raw data of telomere length estimates from Terminal Restriction Fragment Assays (TRF) for various individuals of each species including appropriate details such as age or size and tissue. Telomere length estimates are given in base pairs (bp). In most cases replicate experiments were conducted on groups of samples three times but on a small number of occasions only two replicate experiments were conducted. Further description of the samples can be found in final report of FRDC 2007/033. The arithmetic average for each individual (sample ID) across the two or three replicate experiments is also given. Bidyanus bidyanus (SilverPerch) Two sheets are contained within. a) Comparison of telomere length between different tissues (heart, liver and muscle) within the three year old age class - two replicate experiments were conducted. b) Comparison of telomere length between fish of different but known ages (0.25, 1, 2, and 3 years old) in each of three tissues, heart, liver and muscle – three replicate experiments were conducted per tissue. Haliotis spp (Abalone species) Three species were tested. H. asinina Telomere length was compared in two age classes-11 month and 18 month old abalone using muscle tissue from the foot. Within gel-variation was also estimated using a single sample run three times on one gel (replicate experiment). H. laevigata x H. rubra hybrids Telomere length was compared in three known age classes – two, three and four years old using muscle tissue from the foot. H. rubra Telomere length was compared in a range of different sized abalone using muscle tissue from the foot. Shell size is also given for each abalone Saccostrea glomerata Three sheets are contained within the file. a) Samples came from Moreton Bay Queensland in 2007. Telomere length was compared in two tissues (gill and mantle) of oysters in three age groups (1, 3 and 4 years) b) Samples came from Moreton Bay Queensland in 2009. Telomere length was compared in three age classes using DNA from gill tissue only c) Samples came from Wallis Lake, New South Wales. Telomere length was estimated from whole body minus the shell from 1 year old oysters, gill tissue of 3 age classes (1.5 years, 3 and 4 years), mantle tissue of two age classes (3 and 4 years). Sagmariasus verreauxi (formerly Jasus verreauxi) Telomere length was estimated from abdomen tissue of puerulus, gill and muscle tissue of 3 year old, large and very large size classes of lobsters. Jasus edwardsii Telomere length was measured in two size classes of lobsters- adults of varying sizes using muscle tissue and puerulus using tissues from the abdomen minus the exoskeleton. Metapenaeus macleayi Telomere length was measured in three size classes of school prawns adults. Muscle tissue was used, minus the exoskeleton.

Development of a DNA based aging technique for use in fisheries assessments. Final report, Australian Fisheries Research and Development Corporation Project 2007/033.

The productivity of a fisheries resource can be quantified from estimates of recruitment, individual growth and natural and fisheries-related mortality, assuming the spatial extent of the resource has been quantified and there is minimal immigration or emigration. The sustainability of a fisheries resource is facilitated by management controls such as minimum and maximum size limits and total allowable catch. Minimum size limits are often set to allow individuals the opportunity to reproduce at least once before the chance of capture. Total allowable catches are a proportion of the population biomass, which is estimated based on known reproduction, recruitment, mortality and growth rates. In some fisheries, however, management actions are put in place without quantification of the resource through the stock assessment process. This occurs because species-specific information, for example individual growth, may not be available. In these circumstances, management actions need to be precautionary to protect against future resource collapse, but this often means that the resource is lightly exploited. Consequently, the productivity of the resource is not fully realised. Australia’s most valuable fisheries are invertebrate fisheries (Australian Department of Agriculture Fisheries and Forestry, 2008). For example, Australian fisheries (i.e. excluding aquaculture) production of crustaceans (largely prawns, rock lobster and crab) was 41,000 tonnes in 2006/7, worth $778 million. Production from mollusc (largely abalone, scallops, oysters and squid) fisheries was 39,000 tonnes, worth $502 million. Together, in 2006/7 crustacean and mollusc fisheries represented 58% of the total value of Australian wild fisheries production. Sustainable management of Australia’s invertebrate fisheries is frustrated by the lack of data on species-specific growth rates. This project investigated a new method to estimate age, and hence individual growth rates, in invertebrate fisheries species. The principle behind the new aging method was that telomeres (i.e. DNA end-caps of chromosomes) get shorter as an individual gets older. We studied commercial crustacean and molluscan species. A vertebrate fish species (silver perch, Bidyanus bidyanus) was used as a control to standardise our work against the literature. We found a clear relationship between telomere length and shell size for temperate abalone (Haliotis rubra). Further research is recommended before the method can be implemented to assist management of wildharvested abalone populations. Age needs to be substituted for shell size in the relationship and it needs to be studied for abalone from several regions. This project showed that telomere length declined with increasing age in Sydney rock oysters (Saccostrea glomerata) and was affected by regional variation. A relationship was not apparent between telomere length and age (or size as a surrogate for age) for crustacean species (school prawns, Metapenaeus macleayi; eastern rock lobster, Sagmariasus verreauxi; southern rock lobster, Jasus edwardsii; and spanner crabs, Ranina ranina). For school prawns, there was no difference between telomere length in males and females. Further research is recommended, however, as telomeric DNA from crustaceans was difficult to analyse using the terminal restriction fragment (TRF) assay. Telomere lengths of spanner crabs and lobsters were at the upper limit of resolution of the assay used and results were affected by degradation and possible contamination of telomeric DNA. It is possible that telomere length is an indicator of remaining lifespan in molluscan and crustacean individuals, as suggested for some vertebrate species (e.g. Monaghan, 2010). Among abalone of similar shell size and among lobster pueruli, there was evidence of individuals having significantly longer or shorter telomeres than the group average. At a population level, this may be a surrogate for estimates of future natural mortality, which may have usefulness in the management of those populations. The method used to assay telomere length (terminal restriction fragment assay) performed adequately for most species, but it was too expensive and time-consuming to be considered a useful tool for gathering information for fisheries management. Research on alternative methods is strongly recommended.

Bivalves for the remediation of prawn farm effluent: identification of some potentially useful species in Southern Queensland

Several species of oysters, clams and mussels are currently being used around the world to create extra profits and help remediate waste-waters from mariculture operations. To identify opportunities and potentially suitable species of bivalves for remediation of prawn farm effluent in Australia, recent literature dealing with bivalve filtration is reviewed, and species occurring naturally in a banana prawn, Penaeus (Fenneropenaeus) merguiensis, grow-out pond and effluent streams at the Bribie Island Aquaculture Research Centre (BIARC) were collected, identified and assessed in terms of their tolerance of high silt loadings over 3 months. Three bivalve species predominated in the BIARC case study. These were the mud ark, Anadara trapezia, the rock oyster, Dendostrea folium, and the pearl shell, Pinctada maculata. The mud ark demonstrated the highest tolerance of silt loading (99% survival), followed by pearl shells and rock oysters (88 and 63% survival respectively).

Marking scallops for release and recapture

Objectives : To develop a method to mark hatchery reared saucer scallops to distinguish them from animals derived from wild populations. Outcomes achieved : Juvenile saucer scallop (Amusium balloti) shells have been successfully marked en masse using 3 chemicals, namely alizarin red S, calcein and oxytetracycline (OTC). Considering spat survival, mark quality and mark duration collectively, the most successful chemical was OTC. Scallop spat immersed for three days in 200 or 300 mg L-1 OTC resulted in good mark incorporation and high survival. Tris was an effective means of buffering pH change during OTC treatment, with no apparent adverse effects to the scallops. The marks from OTC treatment were still visible in live scallops for at least 10 months, even with exposure to natural filtered light during that period. A second discernible shell mark was added 27 days after the first with no evident toxicity to the scallops. A simulated seabed system was designed which provide marked improvements in scallop juvenile survival and growth. Advice on shell marking has been given to QSS by DPI&F, and the first commercial trials have now commenced, with initial results showing successful marking of juvenile scallops at QSS. This research will allow the industry to monitor the survival, growth and movement of specific cohorts of deployed scallops. This will provide valuable feedback to assess the value of the ranching venture, to optimise release strategies, and to develop improved species management plans.

Oxides supported carbon air-electrodes for alkaline solution power devices

Porous carbon oxygen-reducing electrodes incorporated with perovskite oxide catalysts are reported. It has been possible to fabricate high-performance oxygen-reducing electrodes by introducing La0.5Sr0.5CoO3 and La0.99Sr0.01NiO3 with the activated coconut-shell charcoal; these electrodes could sustain load currents as high as 1 A cm−2 without serious degradation. A model to explain oxygen-reducing activity of these oxides has been proposed.