A hydrogen peroxide electrode assay to measure thiol peroxidase activity for organoselenium and organotellurium drugs

Molecular mimics of the enzyme glutathione peroxidase (GPx) are increasingly being evaluated as redox active drugs. Their molecular mechanism of action parallels that of the native enzyme; however, a major distinction is that GPx mimics can use alternative thiol substrates to glutathione. This generic thiol peroxidase activity implies that it is necessary to assess a GPx mimic's recognition of a range of cellular thiols in order to determine its potential therapeutic effects. We report an electrochemical assay that, by measuring the rate of decrease of the peroxide substrate, allows the activity of GPx mimics to be directly compared against an array of thiols. The derived pseudo zero-order rate constants, k(obs), for representative GPx mimics range between 0 and 6.6min(-1) and can vary by more than an order of magnitude depending on the thiol electron donor. An additional advantage of the assay is that it enables synergistic interactions between GPx mimics and cellular proteins to be evaluated. Here we report that glutathione disulfide reductase, which is commonly used to evaluate GPx mimic activity, recognizes the GPx mimic ebselen as a substrate, increasing its apparent k(obs). Therefore, reports relying on glutathione disulfide reductase to evaluate GPx mimic activity may exaggerate drug antioxidant action.     

Chromatin domains, insulators, and the regulation of gene expression

The DNA sequence elements called insulators have two basic kinds of properties. Barrier elements block the propagation of heterochromatic structures into adjacent euchromatin. Enhancer blocking elements interfere with interaction between an enhancer and promoter when placed between them. We have dissected a compound insulator element found at the 5' end of the chicken β-globin locus, which possesses both activities. Barrier insulation is mediated by two kinds of DNA binding proteins: USF1/USF2, a heterodimer which recruits multiple enzyme complexes capable of marking histone on adjacent nucleosomes with 'activating' marks, and Vezf1, which protects against DNA methylation. We have found that the heterochromatic region upstream of the insulator element is maintained in its silent state by a dicer-dependent mechanism, suggesting a mechanism for Vezf1 function in the insulator. Enhancer blocking function in the β-globin insulator element is conferred by a binding site for CTCF. Consistent with this property, CTCF binding was found some years ago to be essential for imprinted expression at the Igf2/H19 locus. Work in many laboratories has since demonstrated that CTCF helps stabilize long-range interactions in the nucleus. We have recently shown that in the case of the human insulin locus such an interaction, over a distance of ~300kb, can result in stimulation of a target gene which itself is important for insulin secretion. This article is part of a Special Issue entitled: Chromatin in time and space.     

Exome Capture Reveals ZNF423 and CEP164 Mutations, Linking Renal Ciliopathies to DNA Damage Response Signaling

Nephronophthisis-related ciliopathies (NPHP-RC) are degenerative recessive diseases that affect kidney, retina, and brain. Genetic defects in NPHP gene products that localize to cilia and centrosomes defined them as "ciliopathies." However, disease mechanisms remain poorly understood. Here, we identify by whole-exome resequencing, mutations of MRE11, ZNF423, and CEP164 as causing NPHP-RC. All three genes function within the DNA damage response (DDR) pathway. We demonstrate that, upon induced DNA damage, the NPHP-RC proteins ZNF423, CEP164, and NPHP10 colocalize to nuclear foci positive for TIP60, known to activate ATM at sites?of DNA damage. We show that knockdown of CEP164 or ZNF423 causes sensitivity to DNA damaging agents and that cep164 knockdown in zebrafish results in dysregulated DDR and an NPHP-RC phenotype. Our findings link degenerative diseases of the kidney and retina, disorders of increasing prevalence, to mechanisms of DDR. PAPERFLICK:     

Antinociception of β-D-glucan from Pleurotus pulmonarius is possibly related to protein kinase C inhibition

β-D-Glucan, a polysaccharide isolated from an edible mushroom Pleurotus pulmonarius (Fr.) Quel., presented antinociceptive activity in mice. This study evaluated the involvement of transient receptor potential (TRP) channels and protein kinase C (PKC) on antinociceptive effect of a (1→3),(1→6)-linked β-D-glucan (GL) in mice. Intraperitoneal administration of GL potently inhibited nociceptive responses induced by intraplantar injections of capsaicin, cinnamaldehyde, menthol, acidified saline and phorbol myristate acetate (PMA). Moreover, Western blot analysis revealed that GL treatment also prevented PMA-induced PKC? activation. Collectively, present results demonstrate that GL could constitute an attractive molecule of interest for the development of new analgesic drugs.     

A model of care for familial hypercholesterolaemia: key role for clinical biochemistry

Familial hypercholesterolaemia (FH) is a dominantly inherited disorder present from birth that causes marked elevation in plasma low-density lipoprotein (LDL) cholesterol concentrations and premature coronary heart disease. There are at least 45,000 people with FH in Australia and New Zealand, but most remain unrecognised and those diagnosed remain inadequately treated. To bridge this gap in coronary prevention the FH Australasia Network has developed a model of care for FH. An executive summary of the model of care is presented, with a commentary on its recommendations and the key role of the clinical biochemistry laboratory.     

The effects of a customized over-the-counter mouth guard on neuromuscular force and power production in trained men and women

Although mouth guards were originally designed for injury prevention, even elite athletes are now using performance mouth guards to improve athletic success. Both expensive custom models and over-the-counter models are available, but the efficacy is not well known. Some athletes remain wary of the perceived potential for detriments using a mouth guard to their performance. Thus, the purpose of this study was to examine various physical performance tests when using a mouth guard including a customized over-the-counter mouth guard. Twenty-six trained men (25 ± 4 years; 1.78 ± 0.07 m; 83.3 ± 11.4 kg) and 24 trained women (23 ± 3 years; 1.65 ± 0.08 m; 62.6 ± 7.8 kg) volunteered for the investigation. The subjects completed a familiarization period and then balanced and randomized treatment conditions that included: (a) a customized Power Balance performance mouth guard (PB MG); (b) a regular over the counter boil-and-bite mouth guard (Reg MG); and (c) a no mouth guard (No MG) treatment condition. At each visit, the subjects completed a testing protocol that was sequenced in the following order: sit-and-reach flexibility, medial-lateral balance, visual reaction time, vertical jump, 10-m sprint, bench throw, and plyo press power quotient (3PQ). Heart rate and rating of perceived exertion (RPE) were recorded around the 3PQ. Significance was set at p ≤ 0.05. Expected significant sex differences existed for all power, strength, and speed variables. Bench throw power (watts) and force (newtons) were significantly higher under PB MG than either Reg MG or No MG or in both men and women. The 3PQ power and force production were higher than that for the other 2 treatments for the PB MG for men only. There were no significant differences for treatment conditions in the heart rate or RPE after the 3PQ test. Men were better able to maintain significantly higher 3PQ power production under PB MG treatment condition compared with the other 2 treatment conditions. Rate of power development was significantly higher in men for the vertical jump when using the PB MG compared with that for other treatment conditions in men only. No differences were observed in flexibility, balance, visual reaction time, or sprint time. The PB MG performance mouth guard improves performance of upper-body loaded power exercises in both men and women and lower body power exercise in men without compromising performance on any other performance parameters.     

New species of Teratosphaeria associated with leaf diseases on Corymbia calophylla (Marri)

Corymbia calophylla is an important eucalypt species endemic to the south-west of Western Australia. It is currently in decline across its native habitat for unknown reasons but perhaps due to climate change or to the presence of a recently described canker disease caused by Quambalaria coyrecup. Few fungal leaf pathogens have been described from C. calophylla. The following paper describes two new species of Teratosphaeria from C. calophylla, Teratosphaeria calophylla sp. nov and T. rubidae sp. nov. In addition, a new epitype was designated for T. australiensis.