Coagulation factor XII (FXII) plays a central role in initiating the intrinsic cascade of blood coagulation. Purified recombinant Human Albumin-tagged Infestin-4 (rHA-Infestin-4) is a recently described FXIIa inhibitor that displayed strong anticoagulant activity without compromising haemostasis in several animal models. We pursued detailed in vitro characterisation of rHA-Infestin-4 and demonstrated that it is a competitive inhibitor of FXIIa with slow on and off rate constants for binding (kon=5x10⁵ M⁻¹s⁻¹, koff=6x10⁻⁴ s⁻¹), it can block FXIIa activation of its physiological substrates (plasma prekallikrein and FXI), and it can inhibit ellagic acid-triggered thrombin generation in plasma. Potency and selectivity profiling in enzyme assays suggest that rHA-Infestin-4 is indeed highly potent on FXIIa (IC50=0.3 ± 0.06, 1.5 ± 0.06, 1.2 ± 0.09 nM, for human, rat, and rabbit FXIIa, respectively) with at least >100-fold selectivity against factors IIa, Xa, IXa, XIa, VIIa, and plasma kallikrein in all three species. rHA-Infestin-4 dose-dependently and markedly reduced clot weight in the arteriovenous shunt thrombosis model in rats and rabbits, accompanied with minimal increase in cuticle bleeding times in either species. rHA-Infestin-4 treatment at 5 mg/kg in rabbit resulted in a 13% reduction in ex vivo FXa activity, demonstrating a modest off-target effect. In summary, our findings confirmed and extended previous reports that inhibition of FXIIa by rHA-Infestin-4 can produce strong antithrombotic efficacy while preserving haemostasis. Our comprehensive selectivity profiling, mode of action, and kinetic studies of rHA-Infestin-4 reveal limitations of this molecule and offer new perspectives on any potential effort of discovering novel FXIIa inhibitors.

Alpha-1 acid glycoprotein (AGP) is a significant drug binding acute phase protein that is present in rats. AGP levels areknown to increase during tissue injury, cancer and infection. Accordingly, when determining effective drug ranges and toxicity limits, consideration of drug binding to AGP is essential. However, AGP levels have not been well established during subclinical infections. The goal of this study was to establish a subclinical infection model in rats using AGP as a biomarker. This information could enhance health surveillance, aid in outlier identification, and provide more informed characterization of drug candidates. An initial study (n = 57) was conducted to evaluate AGP in response to various concentrations of Staphylococcus aureus (S. aureus) in Sprague–Dawley rats with or without implants of catheter material. A model validation study (n = 16) was then conducted using propranolol. Rats received vehicle control or S. aureus and when indicated, received oral propranolol (10 mg/kg). Health assessment and blood collection for measurement of plasma AGP or propranolol were performed over time (days). A dose response study showed that plasma AGP was elevated on day 2 in rats inoculated with S. aureus at 106, 107 or, 108 CFU regardless of implant status. Furthermore, AGP levels remained elevated on day 4 in rats inoculated with 107 or 108 CFUs of S. aureus. In contrast, significant increases in AGP were not detected in rats treated with vehicle or 103 CFU S. aureus. In the validation study, robust elevations in plasma AGP were detected on days 2 and 4 in S. aureus infected rats with or without propranolol. The AUC levels for propranolol on days 2 and 4 were 493 ± 44 h × ng/mL and 334 ± 54 h × ng/mL, respectively), whereas in noninfected rats that received only propranolol, levels were 38 ± 11 h × ng/mL and 76 ± 16.h × ng/mL, respectively. The high correlation between plasma propranolol and AGP demonstrated a direct impact of AGP on drug pharmacokinetics and pharmacodynamics. The results indicate that AGP is a reliable biomarker in this model of subclinical infection and should be considered for accurate data interpretation.

No abstract

Government are challenged to deploy trustworthy regulation to enable profitable and environmentally sustainable unconventional petroleum projects. A key activity under scrutiny during the development of these projects is hydraulic fracture stimulation. Regulatory Nirvana for unconventional projects and conventional projects alike entails • Pragmatic licence tenure• Regulatory certainty and efficiency without taint of capture• Regulators and licensees with trustworthy competence and capacity• Effective stakeholder consultation well-ahead of land access• Public access to details of significant risks and reliable research to backup risk management strategies so the basis for regulation is contestable anytime, everywhere• Timely notice of entry with sufficient operational details to effectively inform stakeholders• Potentially affected people and organisations can object to land access -without support for vexatious objections• Fair and expeditious dispute resolution processes• Fair compensation to affected land-users• Risks are reduced to low or as low as reasonably practicable ALARP while also meeting community expectations for net outcomes

If a fraction of the national potential to produce unconventional gas is realised, then Australia will benefit: security of domestic and export gas supplies for decades to come; supply-side competition for decades to come; improved balance of trade and transport fuel security as Australia's supplants imports with gas-based transport fuel; billions of dollars invested in environmentally sustainable projects; thousands of jobs; considerable royalties and tax for revenues public good; and, world-class intellectual property that can be converted into export services and equipment. Given these drivers, the SA State Government convened a Roundtable for Unconventional Gas Projects in October 2010. Participating in this roundtable are a total of 260 organisations plus individuals, including: peak representative bodies focused on economic, social, and natural environment outcomes; and, companies, universities, and key agencies from all state, NT, and commonwealth governments. This roundtable informed a Roadmap for Unconventional Gas Projects in South Australia that was published in December 2012. The objectives of this roadmap are to credibly inform industry strategies, government policies, and public perceptions. In particular, this roadmap explains how people and enterprises potentially affected by unconventional gas operations are given information and time to draw considered views so their rights to object in part or full to activity—and location-specific land access—are supported. This will facilitate the efficient, profitable, and welcomed deployment of capital, technologies, and infrastructure for the commercialisation of unconventional gas. This extended abstract details the findings of this roadmap.