Enzyme-catalysed non-oxidative decarboxylation of aromatic acids: I.Purification and spectroscopic properties of 2,3 dihydroxybenzoic acid decarboxylase from Image Image

In order to understand the molecular mechanism of non-oxidative decarboxylation of aromatic acids observed in microbial systems, 2,3 dihydroxybenzoic acid (DHBA) decarboxylase from Image Image was purified to homogeneity by affinity chromatography. The enzyme (Mr 120 kDa) had four identical subunits (28 kDa each) and was specific for DHBA. It had a pH optimum of 5.2 and Km was 0.34mM. The decarboxylation did not require any cofactors, nor did the enzyme had any pyruvoyl group at the active site. The carboxyl group and hydroxyl group in the Image -position were required for activity. The preliminary spectroscopic properties of the enzyme are also reported.

"Re-thinking" the influence of regulatory capture in the development of government regulation

On 30 March 2015 the Australian Federal Government launched its "Re-Think" initiative with the objective of achieving a better tax system which delivers taxes that are lower, simpler and fairer. The discussion paper released as part of the "Re:think" initiative is designed to start a national conversation on tax reform. However, inquiries into Australia's future tax system, subsequent reforms and the introduction of new taxes are nothing new. Unfortunately, recent history also demonstrates that reform initiatives arising from reviews of the Australian tax system are often deemed a failure. The most prominent of these failures in recent times is the Minerals Resource Rent Tax (MRRT), which lasted a mere 16 months before its announced repeal. Using the established theoretic framework of regulatory capture to interpret publically observable data, the purpose of this article is to explain the failure of this arguably sound tax. It concludes that the MRRT legislation itself, through the capture by the mining companies, provided internal subsidization in the form of reduced tax and minimal or no rents. In doing so, it offers an opportunity to understand and learn from past experiences to ensure that recommendations coming out of the Re:think initiative do not suffer the same fate.

Development of an immunomagnetic capture-reverse transcriptase-PCR assay for three pineapple ampeloviruses

A semi-automated, immunomagneticcapture-reverse transcription PCR(IMC-RT-PCR) assay for the detection of three pineapple-infecting ampeloviruses, Pineapple mealybug wilt-associated virus-1, -2 and -3, is described. The assay was equivalent in sensitivity but more rapid than conventional immunocapture RT-PCR. The assay can be used either as a one- or two-step RT-PCR and allows detection of the viruses separately or together in a triplex assay from fresh, frozen or freeze-dried pineapple leaf tissue. This IMC-RT-PCR assay could be used for high throughput screening of pineapple planting propagules and could easily be modified for the detection of other RNA viruses in a range of plant species, provided suitable antibodies are available.

Mortality and blood loss by blue swimmer crabs (Portunus pelagicus) after simulated capture and discarding from gillnets

Two laboratory experiments were carried out to quantify the mortality and physiological responses of juvenile blue swimmer crabs (Portunus pelagicus) after simulated gillnet entanglement, air exposure, disentanglement, and discarding. In both experiments, all but control blue swimmer crabs were entangled in 1-m(2) gillnet panels for 1 h, exposed to air for 2 min, subjected to various treatments of disentanglement ranging between the forceful removal of none, one, two, and four appendages, then "discarded" into individual experimental tanks and monitored for 10 d. In Experiment 1, mortalities were associated with the number of appendages removed and the occurrence of unsealed wounds. In Experiment 2, live blue swimmer crabs were sampled for blood at 2 min and 6, 24, and 72 h post-discarding to test for the effects of disentanglement and appendage removal on total haemocyte counts, clotting times, protein levels (by refractive index), and blood ion concentrations. Compared with blue swimmer crabs that had sealed or no wounds, those with unsealed wounds had lower total haemocyte counts, protein, and calcium concentrations and increased clotting ties and magnesium and sodium levels. Induced autotomy, as opposed to the arbitrary, forceful removal of a appendages has the potential to minimize the mortality and stress of discarded, juvenile blue swimmer crabs.

Post capture necrosis of the feet caused by soft-catch traps – incidents, cause and prevention.

When recapturing satellite collared wild dogs that had been trapped one month previous in padded foothold traps, we noticed varying degrees of pitting on the pads of their trapped paw. Veterinary advice, based on images taken of the injuries, suggests that the necrosis was caused by vascular compromise. Five of six dingoes we recaptured had varying degrees of necrosis restricted only to the trapped foot and ranging from single 5 mm holes to 25% sections of the toe pads missing or deformed, including loss of nails. The traps used were rubber-padded, two–coiled, Victor Soft Catch #3 traps. The springs are not standard Victor springs but were Beefer springs; these modifications slightly increase trap speed and the jaw pressure on the trapped foot. Despite this modification the spring pressure is still relatively mild in comparison to conventional long spring or four-coiled wild dog traps. The five wild dogs developing necrosis were trapped in November 2006 at 5-6 months of age. Traps were checked each morning so the dogs were unlikely to have been restrained in the trap for more than 12 hours. All dogs exhibited a small degree of paw damage at capture which presented itself as a swollen paw and compression at the capture point. In contrast, eight wild dogs, 7-8 month-old, were captured two months later in February. Upon their release, on advice from a veterinarian, we massaged the trapped foot to get blood flow back in to the foot and applied a bruise treatment (Heparinoid 8.33 mg/ml) to assist restoring blood flow. These animals were subsequently recaptured several months later and showed no signs of necrosis. While post-capture foot injuries are unlikely to be an issue in conventional control programs where the animal is immediately destroyed, caution needs to be used when releasing accidentally captured domestic dogs or research animals captured in rubber-padded traps. We have demonstrated that 7-8 month old dogs can be trapped and released without any evidence of subsequent necrosis following minimal veterinary treatment. We suspect that the rubber padding on traps may increase the tourniquet effect by wrapping around the paw and recommend the evaluation of offset laminated steel jaw traps as an alternative. Offset laminated steel jaw traps have been shown to be relatively humane producing as few foot injuries as rubber-jawed traps.

Assessing the effects of line capture and barotrauma relief procedures on post-release survival of key tropical reef fish species in Australia using recreational tagging clubs.

Common coral trout, Plectropomus leopardus Lacepede, crimson snapper, Lutjanus erythropterus Bloch, saddletail snapper, Lutjanus malabaricus (Bloch & Schneider), red emperor, Lutjanus sebae (Cuvier), redthroat emperor, Lethrinus miniatus (Schneider) and grass emperor, Lethrinus laticaudis Alleyne & Macleay, were tagged to determine the effects of barotrauma relief procedures (weighted shot-line release and venting using a hollow needle) and other factors on survival. Release condition was the most significant factor affecting the subsequent recapture rate of all species. Capture depth was significant in all species apart from L. malabaricus and L. miniatus, the general trend being reduced recapture probability with increasing capture depth. Recapture rates of fish hooked in either the lip or mouth were generally significantly higher than for those hooked in the throat or gut. Statistically significant benefit from treating fish for barotrauma was found in only L. malabaricus, but the lack of any negative effects of treating fish indicated that the practices of venting and shot-lining should not be discouraged by fisheries managers for these species.

Effect of position on precision within the recording field of a markerless motion capture system

Introduction Markerless motion capture systems are relatively new devices that can significantly speed up capturing full body motion. A precision of the assessment of the finger’s position with this type of equipment was evaluated at 17.30 ± 9.56 mm when compare to an active marker system [1]. The Microsoft Kinect was proposed to standardized and enhanced clinical evaluation of patients with hemiplegic cerebral palsy [2]. Markerless motion capture systems have the potential to be used in a clinical setting for movement analysis, as well as for large cohort research. However, the precision of such system needs to be characterized. Global objectives • To assess the precision within the recording field of the markerless motion capture system Openstage 2 (Organic Motion, NY). • To compare the markerless motion capture system with an optoelectric motion capture system with active markers. Specific objectives • To assess the noise of a static body at 13 different location within the recording field of the markerless motion capture system. • To assess the smallest oscillation detected by the markerless motion capture system. • To assess the difference between both systems regarding the body joint angle measurement. Methods Equipment • OpenStage® 2 (Organic Motion, NY) o Markerless motion capture system o 16 video cameras (acquisition rate : 60Hz) o Recording zone : 4m * 5m * 2.4m (depth * width * height) o Provide position and angle of 23 different body segments • VisualeyezTM VZ4000 (PhoeniX Technologies Incorporated, BC) o Optoelectric motion capture system with active markers o 4 trackers system (total of 12 cameras) o Accuracy : 0.5~0.7mm Protocol & Analysis • Static noise: o Motion recording of an humanoid mannequin was done in 13 different locations o RMSE was calculated for each segment in each location • Smallest oscillation detected: o Small oscillations were induced to the humanoid mannequin and motion was recorded until it stopped. o Correlation between the displacement of the head recorded by both systems was measured. A corresponding magnitude was also measured. • Body joints angle: o Body motion was recorded simultaneously with both systems (left side only). o 6 participants (3 females; 32.7 ± 9.4 years old) • Tasks: Walk, Squat, Shoulder flexion & abduction, Elbow flexion, Wrist extension, Pronation / supination (not in results), Head flexion & rotation (not in results), Leg rotation (not in results), Trunk rotation (not in results) o Several body joint angles were measured with both systems. o RMSE was calculated between signals of both systems. Results Conclusion Results show that the Organic Motion markerless system has the potential to be used for assessment of clinical motor symptoms or motor performances However, the following points should be considered: • Precision of the Openstage system varied within the recording field. • Precision is not constant between limb segments. • The error seems to be higher close to the range of motion extremities.

Assessment of the underreporting rate of death cause data in Shandong province using capture-mark-recapture method [应用捕获再捕获方法评估山东省死因登记漏报率水平]

To evaluate the underreporting rate of death -cause data in Shandong province during 2012 to 2013 by capture -mark -recapture method and to provide the base for health strategy. Methods All counties were divided into 5 stratifications according the death rates of 2012, and 14 counties were selected, then 3 towns or streets were selected in each country, 10 villages or neighborhood committees were selected in each town （street）. The death data collected from security bureau and civil affairs bureau were compared with the reporting death data from the National Cause of Death Surveillance, and the underreporting rate was calculated. Results In present study, 6 929 death cases were collected, it was found that 1 556 cases were underreported. The death cases estimated by CMR method were 6 227 cases （95%CI: 7 593-7 651）, and the average underreporting rate was 23.15%. There were significantly differences between different stratifications （P<0.01）. The underreporting rate in 0-4 years old group was 56.93%, the male underreporting rate was 22.31% and the female underreporting rate was 24.09%. There was no significant difference between male and female groups （P>0.05）. Conclusion There is an obvious underreport in the cause of death surveillance of Shandong province, and the underreporting rates are different among the 5 stratifications. The underreporting rate is higher in 0-4 years old group, and the investigation of the death cause surveillance for young residents is not perfect in some countries. The investigation quality of the death cause surveillance should be improved, increasing the integrity of the report data and adjusting the mortalities in different stratifications for obtaining a accurate mortality in Shandong province.

A free-enzyme catalyst for the bioremediation of environmental atrazine contamination.

Herbicide contamination from agriculture is a major issue worldwide, and has been identified as a threat to freshwater and marine environments in the Great Barrier Reef World Heritage Area in Australia. The triazine herbicides are of particular concern because of potential adverse effects, both on photosynthetic organisms and upon vertebrate development. To date a number of bioremediation strategies have been proposed for triazine herbicides, but are unlikely to be implemented due to their reliance upon the release of genetically modified organisms. We propose an alternative strategy using a free-enzyme bioremediant, which is unconstrained by the issues surrounding the use of live organisms. Here we report an initial field trial with an enzyme-based product, demonstrating that the technology is technically capable of remediating water bodies contaminated with the most common triazine herbicide, atrazine.

Identification of amino acid residues essential for enzyme activity of sheep liver 5,10-methylenetetrahydrofolate reductase

Sheep liver 5,10-methylenetetrahydrofolate reductase was subjected to specific chemical modification with phenylglyoxal, diethyl pyrocarbonate and N-bromosuccinimide. The second-order rate constants for inactivation were calculated to be 54 M-1 X min-1, 103 M-1 X min-1 and 154 M-1 X min-1 respectively. This inactivation could be prevented by incubation with substrates or products, suggesting that the residues modified, namely arginine, histidine and tryptophan, are essential for enzyme activity.

A conformational approach to the study of the dynamics of enzyme inhibition: studies on thermolysin

The preferred conformations of β-phenylpropionyl-Image -phenylalanine (β-PPP) and N-carbobenzoxy-L-phenylalanine (Cbz-Phe), two inhibitors of thermolysin, have been determined by computing potential energy using empirial potential energy functions. Of the 15 to 20 conformations that are favoured for each of these inhibitors only a few have the right conformation to reach the active site of the enzyme. The conformer of β-PPP that initiates binding with the enzyme is different from the bound one, while for Cbz-Phe the bound and initiating conformers are quite similar. Thus, β-PPP favours the ‘induced fit’ model while Cbz-Phe follows the ‘lock and key’ model of binding. The inhibitors differ in their alignment at the active site.

Effect of configuration of the inhibitors on the mode of binding to the enzyme, thermolysin

Preferred conformations of the competitive inhibitors glycyl-L-phenylalanine and glycyl-D-phenylalanine and their mode of binding to thermolysin have been studied. The difference in configuration is shown to affect significantly the mode of binding to thermolysin. Gly-D-Phe prefers to enter the active site in the global minimum conformation whereas Gly-L-Phe may enter in a higher energy conformation. Moreover, D-enantiomer is shown to have a better fit than the L-counterpart in the active site.

Studies on immune activation in rheumatoid arthritis and reactive arthritis

Rheumatoid arthritis (RA) is an autoimmune disease characterized by synovitis, progressive joint destruction, and disability. Reactive arthritis (ReA) is a sterile joint inflammation following a distant mucosal infection. The clinical course of these diseases is variable and cannot be predicted with reasonable accuracy by clinical and laboratory markers. The predictive value of circulating soluble interleukin-2 receptor (sIL-2R), a marker of lymphocyte activation, measured by Immulite® automated immunoassay analyzer, was evaluated in two cohorts of RA patients. In 175 patients with active early RA randomized to treatment with either on disease-modifying antirheumatic drug (DMARD) or a combination of 3 DMARDs and prednisolone, low baseline sIL-2R level predicted remission after 6 months in patients treated with a single DMARD. In 24 patients with active RA refractory to DMARDs, low baseline sIL-2R level predicted rapid clinical response to treatment with infliximab, an anti-tumour necrosis factor antibody. Furthermore, in a cohort of 26 patients with acute ReA, high baseline sIL-2R level predicted remission after 6 months. Levels of circulating soluble E-selectin (sE-selectin), a marker of endothelial activation, were measured annually by enzyme-linked immunosorbent assay (ELISA) in a cohort of 85 patients with early RA. During a five-year follow-up, sE-selectin levels were associated with activity and outcome of RA. The levels of neutrophil and monocyte CD11b/CD18 expression measured by flow cytometry, and circulating levels of sE-selectin measured by ELISA, and procalcitonin by immunoluminometric assay, were compared in 28 patients with acute ReA and 16 patients with early RA. The levels of the markers were comparable in ReA, RA, and healthy control subjects. In conlusion, sIL-2R may provide a new predictive marker in early RA treated with a single DMARD and refractory RA treated with infliximab. In addition, sIL-2R level predicts remission in acute ReA.

A core metabolic enzyme mediates resistance to phosphine gas

Phosphine is a small redox-active gas that is used to protect global grain reserves, which are threatened by the emergence of phosphine resistance in pest insects. We find that polymorphisms responsible for genetic resistance cluster around the redox-active catalytic disulfide or the dimerization interface of dihydrolipoamide dehydrogenase (DLD) in insects (Rhyzopertha dominica and Tribolium castaneum) and nematodes (Caenorhabditis elegans). DLD is a core metabolic enzyme representing a new class of resistance factor for a redox-active metabolic toxin. It participates in four key steps of core metabolism, and metabolite profiles indicate that phosphine exposure in mutant and wild-type animals affects these steps differently. Mutation of DLD in C. elegans increases arsenite sensitivity. This specific vulnerability may be exploited to control phosphine-resistant insects and safeguard food security.

Carbon Capture and Productivity of Plantation Forests in North-eastern Australia

Approximately 130,000 ha of hardwood plantations have been established in north-eastern Australia in the last 15 years. As a result of poor taxa selection approximately 25,000 ha have failed due to drought, pest and disease or extreme weather events (drought and cyclones). Given the predicted impacts of climate change in north-eastern Australia (reduced rainfall, increased temperatures and an increase in extreme weather conditions, particularly drought, storms and cyclones), selection of the right taxa for plantation development is even more critical as the taxon planted needs to be able to perform well under the environments experienced at planting as well as those that may develop over in 30 years time as a result of changing climate.