MPO (myeloperoxidase) catalyses the oxidation of chloride, bromide and thiocyanate by hydrogen peroxide to HOCl (hypochlorous acid), HOBr (hypobromous acid) and HOSCN (hypothiocyanous acid) respectively. Specificity constants indicate that SCN− is a major substrate for MPO. HOSCN is also a major oxidant generated by other peroxidases including salivary, gastric and eosinophil peroxidases. While HOCl and HOBr are powerful oxidizing agents, HOSCN is a less reactive, but more specific, oxidant which targets thiols and especially low pKa species. In the present study we show that HOSCN targets cysteine residues present in PTPs (protein tyrosine phosphatases) with this resulting in a loss of PTP activity for the isolated enzyme, in cell lysates and intact J774A.1 macrophage-like cells. Inhibition also occurs with MPO-generated HOCl and HOBr, but is more marked with MPO-generated HOSCN, particularly at longer incubation times. This inhibition is reversed by dithiothreitol, particularly at early time points, consistent with the reversible oxidation of the active site cysteine residue to give either a cysteine–SCN adduct or a sulfenic acid. Inhibition of PTP activity is associated with increased phosphorylation of p38a and ERK2 (extracellular-signal-regulated kinase 2) as detected by Western blot analysis and phosphoprotein arrays, and results in altered MAPK (mitogen-activated protein kinase) signalling. These data indicate that the highly selective targeting of some protein thiols by HOSCN can result in perturbation of cellular phosphorylation and altered cell signalling. These changes occur with (patho)physiological concentrations of SCN− ions, and implicate HOSCN as an important mediator of inflammation-induced oxidative damage, particularly in smokers who have elevated plasma levels of SCN−.

Comparison of seal oil to tuna oil on plasma lipid levels and blood pressure in hypertriglyceridaemic subjects AbstractAs meat is a rich source of the omega-3 fatty acid docosapentaenoic acid (DPA) and Australians consume six times more meat than fish, investigation of the potential health benefit of DPA is warranted. The aims were to compare the effects of seal oil supplementation with fish oil, on measures of plasma lipids and blood pressure in hypertriglyceridaemic subjects. Forty-eight volunteers were recruited from the Wollongong community and were randomly allocated to one of three groups either receiving 1 g/day of long-chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) using one of three oils: seal oil capsules (340 mg eicosapentaenoic acid (EPA), 230 mg DPA, 450 mg DHA), fish oil capsules (210 mg EPA, 30 mg DPA, 810 mg DHA) or placebo capsules (containing sunola oil) for 6 weeks. Plasma triglycerides remained unchanged in the placebo group, whilst reductions of 7 and 14% (P\0.05) were seen in the fish oil and seal oil groups respectively. Systolic blood pressure improved by 8 and 5 mmHg with seal oil and fish oil respectively (P\0.05). The mean arterial pressure was significantly lower after seal oil supplementation (P\0.005) compared with the placebo group. These results indicate that seal oil is as effective as fish oil in lowering plasma triglycerides and blood pressure. AbstractAs meat is a rich source of the omega-3 fatty acid docosapentaenoic acid (DPA) and Australians consume 6 times more meat than fish, investigation of the potential health benefit of DPA is warranted. The aims were to compare the effects of seal oil supplementation with fish oil, on measures of plasma lipids and blood pressure in hypertriglyceridaemic subjects. Forty-eight volunteers were recruited from the Wollongong community and were randomly allocated to one of three groups receiving 1g/d of long chain omega-3 polyunsaturated fatty acids (LC n-3 PUFA) using one of three oils: seal oil capsules (340mg eicosapentaenoic acid (EPA), 230mg DPA, 450mg DHA), fish oil capsules (210mg EPA, 30mg DPA, 810mg DHA) or placebo capsules (containing sunola oil) for 6 weeks. Plasma triglycerides remained unchanged in the placebo group, whilst reductions of 7% and 14% (p<0.05) were seen in the fish oil and seal oil groups respectively. Systolic blood pressure improved by 8 and 5 mmHg with seal oil and fish oil respectively (p<0.05).The mean arterial pressure was significantly lower after seal oil supplementation (p<0.005) compared with the placebo group. These results indicate that seal oil is as effective as fish oil in lowering plasma triglycerides and blood pressure.

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Four novel, disubstituted diaminopteridines have been identified which antagonize the uptake of a folate precursor (para-aminobenzoic acid) by rat-derived Pneumocystis carinii maintained in short-term axenic culture at concentrations ranging from 4.5 to 26 microM. The compounds were at least 10 to 100 times more active than trimethoprim in this assay. None of these entities exhibited toxicity to mammalian cell lines at < 100 microM. The same structures also caused significant inhibition of Toxoplasma gondii tachyzoite replication within Madin-Darby bovine kidney cells at concentrations ranging from 0.1 to 10 microM. Three of the structures (GR92754, AH10639, and AH2504) were at least an order of magnitude more potent than the standard anti-T. gondii agent, pyrimethamine. All three entities were also significantly more potent and selective than pyrimethamine as inhibitors of T. gondii dihydrofolate reductase (DHFR), with 50% inhibitory concentrations within the range of 0.018 to 0.033 microM. One of these compounds, 6,7-dibutyl-2,4-diaminopteridine (GR92754), was also a potent and selective inhibitor of P. carinii DHFR (50% inhibitory concentration, 0.082 microM). GR92754 is the first DHFR inhibitor described that exhibits greater potency, selectivity, and intracellular activity against both organisms than any of the DHFR agents used clinically, namely, trimethoprim, pyrimethamine, and trimetrexate. This information could provide the starting point for examination of the pharmacokinetic and therapeutic potential of GR92754 and related chemical entities with animal models.