Probing the microenvironmental conditions for induction of superficial zone protein expression

SummaryObjectiveTo determine the in vitro conditions which promote expression of superficial zone protein (SZP).MethodsChondrocytes from 6-month-old calves were expanded in monolayer culture and the expression of SZP in alginate bead and monolayer culture was quantified with quantitative real time-polymerase chain reaction (qRT-PCR) and immunostaining. The effect of oxygen tension on SZP expression was determined by qRT-PRC analysis of cells cultured in two dimension (2D) and three dimension (3D) under hypoxic (1% pO2) or normoxic (21% pO2) conditions. Finally, to examine the effect of cyclic tensile strain on expression of SZP in 2D and 3D cultures, chondrocytes encapsulated in alginate beams or seeded on type I collagen coated polydimethylsiloxane (PDMS) chambers were subjected to 5% strain at 1 Hz, 2 h/day for 4 days or 2 h at the fourth day of culture and mRNA levels were quantified.ResultsBovine chondrocytes in monolayer showed a drastic decrease in SZP expression, similar in trend to the commonly reported downregulation of type II collagen (Col2). Chondrocytes embedded in alginate beads for 4 days re-expressed SZP but not Col2. SZP expression was higher under normoxic conditions whereas Col2 was upregulated only in alginate beads under hypoxic conditions. Cyclic mechanical strain showed a tendency to upregulate mRNA levels of SZP.ConclusionsA microenvironment encompassing a soft encapsulation material and 21% oxygen is sufficient for fibroblastic chondrocytes to re-express SZP. These results serve as a guideline for the design of stratified engineered articular cartilage and suggest that microenvironmental cues (oxygen tension level) strongly influence the pattern of SZP expression in vivo

Sheep Updates 2009

This session covers seven papers from different authors: 1. Scouring Management and Worm Control, Brown Besier, Department of Agriculture and Food, Western Australia 2.Breeding sheep for resistance to breech strike:- Selection results in WA, LJE Karlsson, JC Greeff & AC Schlink, Department of Agriculture and Food, Western Australia 3.Future Ewe - matching genetics to the production system, Mark Ferguson, Department of Agriculture and Food, Western Australia 4. Within-flock selection of ewes: opportunities for gains in reproduction, Greg Leeand Sue Hatcher, NSW Department of Primary Industries & Australian CRCforSheep Industry Innovation (Orange) 5. Managing Merinos on Murrayfield, Bruce Michael, Murryfield, Bruny Island, Tasmania 6. Managing [breech] flystrike in [unmulesed] sheep, Rob Woodgate, Darren Michael, Mandy Curnow and Julia Smith, Department of Agriculture and Food, Western Australia 7. Value of Pregnancy Scanning and Differential Feeding of Dry, Single amd Twin Ewes, John Young, Farming Systems Analysis Service, Kojonup, WA, Andrew Thompson, Chris Oldham Department of Agriculture and Food, Western Australi

Sheep Updates 2003 - Posters

This session covers eleven papers from different authors:1 Sheep production on annual stubbles/pastures vs lucerne Maxine Brown Gaye Krebs Muresk Institute, Curtin University Diana Fedorenko Kathryn Egerton-Warburton Centre for Cropping Systems, Department of Agriculture Western Australia 2. The value chain of the Lake Grace livestock industry Evan Burt Nazrul Islam Department of Agriculture Western Australia 3. Native pastures, Dorper sheep and the 2002 drought Roy Butler Department of Agriculture Western Australia 4. Commercial sheep breeders can improve their sheep breeding program using wether trials L.G. Butler, S.R. Brown, M.F. D’Antuono, J.C. Greeff Department of Agriculture 5. Western Australia Linked ewe trials to benchmark wool traits and reproductive performance of Western Australian sheep flocks Ken Hart Department of Agriculture Western Australia 6. Damara sheep - what is their potential? A case study from the North-eastern wheatbelt Tanya Kilminster Evan Burt Department of Agriculture Western Australia 7, Australian Sheep Industry CRC - nutrition sub-program Rachel Kirby Sheep CRC Research Fellow 8. Dust penetration is not genetically and phenotypically the same trait as dust content M.E. Ladyman J.C. Greeff Department of Agriculture Western Australia A.C. Schlink CSIRO Livestock Industries, Private Bag 5, Wembley WA I.H. Williams P.E. Vercoe University of Western Australia, Crawley WA 9.Developing sustainable fodder crop systems with new annual pasture legumes Anyou Lui Department of Agriculture Western Australia 10. Seasonal pricing and seasonality of supply of prime lambs in the western wheatbelt Karen Smith Martin Bent Muresk Institute, Curtin University 11. The role of alternative and exotic sheep breeds in the Western Australian sheep industry Matthew Young Department of Agriculture Western Australi

Agribusiness Sheep Updates - 2004 - Part 1

Proceedings of the Agribusiness Sheep Updates - 2004 Forward Dr Mark Dolling Manager, Sheep Industries and Pasture, Department of Agriculture Western Australia Keynotes Australian Wool Innovation Limited DR LEN STEPHENS AUSTRALIAN WOOL INNOVATION LIMITED (AWI) Commercialisation of Sheepmeat Eating Quality Outcomes, David Thomason, General Manger Marketing Meat & livestock Australia Limited PLENARY The Fitness of the Future Merino, Norm Adams and Shimin Liu, CSIRO Livestock Industries Ovine Johne’s Disease – Managing the Disease, Managing the Issues, PETER BUCKMAN, CHIEF VETERINARY OFFICER, DEPARTMENT OF AGRICULTURE WESTERN AUSTRALIA Animal Welfare – Changes in Latitudes Changes in Attitudes, Michael Paton and Dianne Evans, Department of Agriculture Western Australian. Live Sheep Exports, JOHN EDWARDS. CHAIRMAN, WESTERN AUSTRALIAN LIVE SHEEP EXPORTERS ASSOCIATION MeCustomising to the Needs of the Customer – Insights from the New Zealand Merino Experience, DR SCOTT CHAMPION, RESEARCH, DEVELOPMENT AND PRODUCT INNOVATION MANAGER, THE NEW ZEALAND MERINO COMPANY LIMITED Agribusiness Sheep Updates Conference -Economic and Financial Market Update Alan Langford, Economist, BankWest Concurrent sessions - Meeting the Market Breeding Wool to Address Consumer Requirements in Fabrics A.C. SCHLINK CSIRO Livestock Industries, J.C. GREEFF AND M. E. LADYMAN Department of Agriculture Western Australia Fibre Contribution to Retail Demand for Knitwear Melanie LadymanA and John StantonAB ADepartment of Agriculture Western Australia and BCurtin University of Technology Sustainable Merino, is this the Future for Merino? Stuart Adams, iZWool International P/L Meeting lamb Market Specs from Crossbred Ewes Dr. Neal Fogarty, NSW Agriculture and the Australian Sheep Industry CRC Use of Serial Body Weight Measurements in Prime Lamb Finishing Systems Matthew Kelly, CSIRO Livestock Industries, James Skerritt, Ian McFarland Department of Agriculture Western Australia, Australian Sheep Industry CR

Sheep Updates 2005 - Part 4

This session covers twelve papers from different authors: REPRODUCTION 1. Is it worth increasing investment to increase lambing percentages? Lucy Anderton Department of Agriculture Western Australia. 2. What value is a lamb? John Young, Farming Systems Analysis Service, Kojonup, WA 3. Providing twin-bearing ewes with extra energy at lambing produces heavier lambs at marking. Rob Davidson WAMMCO International,, formerly University of Western Australia; Keith Croker, Ken Hart, Department of Agriculture Western Australia, Tim Wiese, Chuckem , Highbury, Western Australia. GENETICS 4. Underlying biological cause of trade-off between meat and wool. Part 1. Wool and muscle glycogen, BM Thomson, I Williams, University of WA, Crawley, JRBriegel, CSIRO Livestock Industries, Floreat Park WA &CRC for the Australian Sheep Industry, JC Greeff, Department of Agriculture Western Australia &CRC for the Australian Sheep Industry. 5. Underlying biological cause of trade-off between meat and wool. Part 2. Wool and fatness, NR Adams1,3, EN Bermingham1,3, JR Briegel1,3, JC Greeff2,3 1CSIRO Livestock Industries, Floreat Park WA 2Department of Agriculture Western Australia, 3CRC for the Australian Sheep Industry 6. Genetic trade-offs between lamb and wool production in Merino breeding programs, Johan Greeff, Department of Agriculture, Western Australia. 7. Clean fleece weight is no phenotypically independent of other traits. Sue Hatcherac and Gordon Refshaugebc aNSWDPI Orange Agricultural Institute, Orange NSW 2800 bUNE c/- NSWDPI Cowra AR&AS Cowra NSW 2794 cAustralian Sheep Industry CRC. 8. When you\u27re on a good thing, do it better: An economic analysis of sheep breed profitability. Emma Kopke, Ross Kingwell, Department of Agriculture, Western Australia, John Young, Farming Systems Analysis Service, Kojonup, WA. 9. Selection Demonstration Flocks: Demonstrating improvementsin productivity of merinos, K.E. Kemper, M.L. Hebart, F.D. Brien, K.S. Jaensch, R.J. Grimson, D.H. Smith South Australian Research and Development Institute 10. You are compromising yield by using Dust Penetration and GFW in breeding programs, Melanie Dowling, Department of Agriculture, Western Australia, A. (Tony) Schlink, CSIRO Livestock Industries, Wembley, Johan Greeff, Department of Agriculture Western Australia. 11. Merino Sheep can be bred for resistance to breech strike. Johan Greeff , John Karlsson, Department of Agriculture Western Australia 12. Parasite resistant sheep and hypersensitivity diarrhoea, L.J.E. Karlsson & J.C. Greeff, Department of Agriculture Western Australi

Sheep Updates 2005 - Part 7

This session covers eight papers from different authors: POSTERS 1. Sulpher in wool and its implications for fleece weight and sheep health, SM Liu, AC Schlink, JR Williams, CSIRO Livestock Industries Wembley WA, ME Dowling,JCGreef, Department of Agriculture Western Australia. 2. Stubbles for sheep: a reality check, Roy Butler, Keith Croker, Department of Agriculture Western Australia. 3. Genetic benchmarking using artificial insemination, LC Butler, JC Greeff, Department of Agriculture Western Australia. 4. The potential lambing performances of ewes in mixed age flocks, Kieth Croker, Department of Agriculture Western Australia, Rob Davidson, WAMMCO International, formally University of Western Australia, Ken Hart, Department of Agriculture Western Australia,Doug Harrington Cowcher Farms Narrogin, Mario D\u27Antuono, Department of Agriculture Western Australia. 5. National Livestock Identification System (Sheep) in Western Australia, Julian Gardner, Department of Agriculture Western Australia. DISPLAYS - TOOLS 6. To Feed or Not to Feed - I Only Hamlet had the Calculator!, Geoff Duddy, Livestock Officer(Sheep & Wool) Yanco. 7. WormBoss - a national Australian computer-based sheep worm control tool, RG Woodgate, Department of Agriculture Western Australia, A LeFeuvre, Queensland Department of Primary Industries and Fisheries, and Genie Pty Ltd, Warwick Qld, A Bailey, Department of Primary Industries, Water and Environment, Kings Meadow Tas, RB Besier, Department of Agriculture Western Australia, N. Campbell, Department of Primary Industries Victoria, Attwood Vic, I Carmichael, South Australian Research and Development Institute, Glenside SA, S. Love, NSW Department of Primary Industries, Armidale NSW. 8. \u27Eye in the sky\u27 takes guesswork out of farmers pasture decisions, Richard Stovold, Department of Land Informatio

Proteolytic enzyme engineering : a tool for wool

One of the goals of protein engineering is to tailor the structure of enzymes to optimize industrial bioprocesses. In the present work, we present the construction of a novel high molecular weight subtilisin, based on the fusion of the DNA sequences coding for Bacillus subtilis prosubtilisin E and for an elastin-like polymer (ELP). The resulting fusion protein was biologically produced in Escherichia coli, purified and used for wool finishing assays. When compared to the commercial protease Esperase, the recombinant subtilisinE-VPAVG220 activity was restricted to the cuticle of wool, allowing a significant reduction of pilling, weight loss and tensile strength loss of wool fibers. Here we report, for the first time, the microbial production of a functionalized high molecular weight protease for controlled enzymatic hydrolysis of wool surface. This original process overcomes the unrestrained diffusion and extended fiber damage which are the major obstacles for the use of proteases for wool finishing applications

Balancing, Proportionality, and Constitutional Rights

In the theory and practice of constitutional adjudication, proportionality review plays a crucial role. At a theoretical level, it lies at core of the debate on rights adjudication; in judicial practice, it is a widespread decision-making model characterizing the action of constitutional, supra-national and international courts. Despite its circulation and centrality in contemporary legal discourse, proportionality in rights-adjudication is still extremely controversial. It raises normative questions—concerning its justification and limits—and descriptive questions—regarding its nature and distinctive features. The chapter addresses both orders of questions. Part I centres on the justification of proportionality review, the connection between proportionality, balancing and theories of rights and the critical aspects of this connection. Part II identifies and analyses the different forms of proportionality both in review, as a template for rights-adjudication, and of review, as a way of defining the scope and limits of adjudication

Current European Labyrinthula zosterae Are Not Virulent and Modulate Seagrass (Zostera marina) Defense Gene Expression

Pro- and eukaryotic microbes associated with multi-cellular organisms are receiving increasing attention as a driving factor in ecosystems. Endophytes in plants can change host performance by altering nutrient uptake, secondary metabolite production or defense mechanisms. Recent studies detected widespread prevalence of Labyrinthula zosterae in European Zostera marina meadows, a protist that allegedly caused a massive amphi-Atlantic seagrass die-off event in the 1930's, while showing only limited virulence today. As a limiting factor for pathogenicity, we investigated genotype×genotype interactions of host and pathogen from different regions (10–100 km-scale) through reciprocal infection. Although the endophyte rapidly infected Z. marina, we found little evidence that Z. marina was negatively impacted by L. zosterae. Instead Z. marina showed enhanced leaf growth and kept endophyte abundance low. Moreover, we found almost no interaction of protist×eelgrass-origin on different parameters of L. zosterae virulence/Z. marina performance, and also no increase in mortality after experimental infection. In a target gene approach, we identified a significant down-regulation in the expression of 6/11 genes from the defense cascade of Z. marina after real-time quantitative PCR, revealing strong immune modulation of the host's defense by a potential parasite for the first time in a marine plant. Nevertheless, one gene involved in phenol synthesis was strongly up-regulated, indicating that Z. marina plants were probably able to control the level of infection. There was no change in expression in a general stress indicator gene (HSP70). Mean L. zosterae abundances decreased below 10% after 16 days of experimental runtime. We conclude that under non-stress conditions L. zosterae infection in the study region is not associated with substantial virulence