Furin proteolytically processes the heparin-binding region of extracellular superoxide dismutase

Extracellular superoxide dismutase (EC-SOD) is an antioxidant enzyme that attenuates brain and lung injury from oxidative stress. A polybasic region in the carboxyl terminus distinguishes EC-SOD from other superoxide dismutases and determines EC-SOD's tissue half-life and affinity for heparin. There are two types of EC-SOD that differ based on the presence or absence of this heparin-binding region. It has recently been shown that proteolytic removal of the heparin-binding region is an intracellular event (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). By using mammalian cell lines, we have now determined that removal of the heparin-binding region occurs after passage through the Golgi network but before being secreted into the extracellular space. Specific protease inhibitors and overexpression of intracellular proteases implicate furin as a processing protease. In vitro experiments using furin and purified EC-SOD suggest that furin proteolytically cleaves EC-SOD in the middle of the polybasic region and then requires an additional carboxypeptidase to remove the remaining lysines and arginines. A mutation in Arg(213) renders EC-SOD resistant to furin processing. These results indicate that furin-dependent processing of EC-SOD is important for determining the tissue distribution and half-life of EC-SOD.     

The intracellular proteolytic processing of extracellular superoxide dismutase (EC-SOD) is a two-step event

Extracellular superoxide dismutase (EC-SOD) is a tetramer composed of either intact (Trp(1)-Ala(222)) or proteolytically cleaved (Trp(1)-Glu(209)) subunits. The latter form is processed intracellularly before secretion and lacks the C-terminal extracellular matrix (ECM)-binding region ((210)RKKRRRESECKAA(222)-COOH). We have previously suggested that the C-terminal processing of EC-SOD is either a one-step mechanism accomplished by a single intracellular endoproteolytic event cleaving the Glu(209)-Arg(210) peptide bond or a two-step mechanism involving two proteinases (Enghild, J. J., Thogersen, I. B., Oury, T. D., Valnickova, Z., Hojrup, P., and Crapo, J. D. (1999) J. Biol. Chem. 274, 14818-14822). In the latter case, an initial endoproteinase cleavage occurs somewhere in the region between Glu(209) and Glu(216). A carboxypeptidase specific for basic amino acid residues subsequently trims the remaining basic amino acid residues to Glu(209). A naturally occurring mutation of EC-SOD substituting Arg(213) for Gly enabled us to test these hypotheses. The mutation does not prevent proteolysis of the ECM-binding region but prevents a carboxypeptidase B-like enzyme from trimming residues beyond Gly(213). The R213G mutation is located in the ECM-binding region, and individuals carrying this mutation have an increased concentration of EC-SOD in the circulatory system. In this study, we purified the R213G EC-SOD variant from heterozygous or homozygous individuals and determined the C-terminal residue of the processed subunit to be Gly(213). This finding supports the two-step processing mechanism and indicates that the R213G mutation does not disturb the initial endoproteinase cleavage event but perturbs the subsequent trimming of the C terminus.     

Antimicrobial susceptibility of Francisella noatunensis subsp. noatunensis strains isolated from Atlantic cod Gadus morhua in Norway

A total of 30 isolates of Francisella noatunensis subsp. noatunensis isolated from Atlantic cod Gadus morhua L. were tested for susceptibility, in the form of minimal inhibitory concentration (MIC) values, against the following antibacterial agents: flumequine, oxolinic acid, ciprofloxacin, florfenicol, oxytetracycline, erythromycin, streptomycin sulphate, trimetoprim/sulphadiazine and rifampin. All the isolates had a low susceptibility to oxytetracycline, trimetoprim/sulphadiazine (Tribrissen?), erythromycin, ciprofloxacin and streptomycin with MIC values of 64, 64 to 128, 16, 8 and 32 to 128 μg ml-1, respectively. The strains were, on the other hand, susceptible to florfenicol, oxolinic acid, flumequine and rifampin with MIC values of 0.5, 0.25, 0.25 and 0.25 to 1 μg ml-1, respectively.     

Inflammatory Chemokines Direct and Restrict Leukocyte Migration within Live Tissues as Glycan-Bound Gradients

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Highlights? Chemokines form tissue-bound gradients in vivo through heparan sulfate proteoglycans ? Chemokines guide neutrophils in vivo as haptotactic gradients ? Chemokines guide neutrophils toward infection loci by triggering orthotaxis ? Chemokines locally restrict neutrophil motility at sites of infection     

Effects of methionine containing paracetamol formulation on serum vitamins and trace elements in male rats

Methionine is an effective antidote in the treatment of paracetamol-induced toxicity but at large doses it has been reported to induce or aggravate a number of pathological conditions. It also alters plasma levels of many vital elements and molecules. This study was designed to identify if the alteration observed for antioxidant vitamins and minerals especially at sub-toxic and toxic levels of exposure in our earlier study of 24-hour exposure period may warrant trace elements supplementation. This was investigated by carrying out a 48-hour study to test the ability of a living organism to restore homeostasis of these vital molecules and elements. The levels of antioxidant minerals and vitamins were estimated in the serum samples obtained from adult male Wistar rats exposed to paracetamol tablets. At 100 mg\kg BW (body weight) vitamin A, niacin, riboflavin, selenium and manganese were not significantly different from the control group. Moreover at 350 mg\kg, all these indices except zinc were not significantly different in the exposed group compared with controls whereas at 1000 mg\kg level of exposure manganese, selenium and vitamin E were not significantly decreased at the end of 48 hours of exposure but copper, niacin and vitamin A were significantly increased in the exposed group compared with the controls. These results suggest that with time the body may be capable of bringing about restoration of the levels of some of these elements\vitamins. This was more evident at 350 mg\kg level of exposure than a higher dose of 1000 mg\kg level.Keywords: Paracetamol formulation, Vitamins, Trace elements.     

Stictyosiphon arcticus (Fucophyceae, Stictyosiphonaceae) a new species from Greenland, North Norway, and Baffin Island

A new species of Stictyosiphon from the North Atlantic is decribeci. Morphologically it resembles a densely branched Chordaria flagelliformis or an aberrant Dictyosiphon foeniculaceus. It disagrees with known species of Stictyosiphon by a loosely constructed cortex which develops in connection with formation of plurilocular sporangia. We, therefore, consider this a new species for which the name Stictyosiphon arcticus is proposed.     

A New Technique for Demonstration of Trichinella Spiralis Larvae in Suspensions of Digested Muscle Tissue

Up to now, a sedimentation technique has been used for demonstration of Trichinella spiralis larvae released by artificial digestion (Henriksen 1973, Skovgaard 1975, Thomsen 1976, 1977, and others).