Alkaline protease production by an actinomycete MA1-1 isolated from marine sediments

Alkaliphilic actinomycetes isolated from sediment samples of the Izmir Gulf, Turkey were studied for the production of protease activity. Strain MA1-1 was selected as a good alkaline protease producer as measured by the clear zone diameter by the hydrolysis of skim-milk and casein. The alkaline protease production from the marine alkaliphilic actinomycete MA1-1 was studied by using different carbon and nitrogen sources in medium containing glycerol, peptone, KCl, MgSO₄, K₂HPO₄, and trace elements at 30°C for 72 h. Among the different carbon and nitrogen sources, fructose, starch, maltose, D(+) glucose, yeast extract, malt extract, beef extract and peptone provided higher production of protease. Starch was also found to be effective for growth and enzyme production with highest specific activity at 699 U mg^?1. Purification was achieved by adsorption on Diaion HP 20 which resulted in a recovery rate of 68% with a specific activity of 7618 U mg^?1 protein and 40-fold purification. The optimum pH and temperature of the partially purified protease were determined as pH 9.0 and 50°C, but high activity was also observed at pH 8.0–13.0 and 35–50°C. The inhibition profile exhibited by phenylmethylsulphonyl fluoride (PMSF) showed that this enzyme belongs to the serine-protease group.     

Distribution and antibacterial drug resistance ofAeromonas spp. from fresh and brackish waters in Southern Turkey

The frequency of antibiotic resistance was compared inAeromonas spp. isolated from fresh and brackish water in Southern Turkey. A total of 97Aeromonas spp. strains were isolated from four zones (three from fresh and one from brackish water). Most of the strains isolated from all samples wereAeromonas hydrophila (79.4%), while the amount ofAeromonas sobria andAeromons caviae, were rather lower in the samples examined (17.5% and 3.1% respectively). A high proportion of isolates from all water sources showed resistance to cephalotin (86.6%) and trimethoprim-sulphamethoxazole (69%). On the other hand, a low proportion of bacteria showed resistance to tetracycline (14.4%), chloramphenicol (11.3%), gentamicin (7.2%) and nitrofurantoin (6.8%). Only one strain showing resistance to amikacin was found. Multiple Antibiotic Resistance Index (MARI) to at least two antibiotics was highest in brackish water (zone 4), followed by fresh water (zone 3). MARI values ranging from 0.2 to 0.8 for the bacteria isolated from brackish water. This study suggest that, multiple antibiotic resistantAeromonas spp., especiallyA. hydrophila, can be easily recovered from fresh and brackish water sources in Turkey and these sources may play as a reservoirs responsible for disease pathogen aeromonads.     

Recovery from hypoxia-induced internalization of cardiac Na+/H + exchanger 1 requires an adequate intracellular store of antioxidants

The heart is highly active metabolically but relatively underperfused and, therefore, vulnerable to ischemia. In addition to acidosis, a key component of ischemia is hypoxia that can modulate gene expression and protein function as part of an adaptive or even maladaptive response. Here, using cardiac-derived HL-1 cells, we investigate the effect of various hypoxic stimuli on the expression and activity of Na⁺/H ⁺ exchanger 1 (NHE1), a principal regulator of intracellular pH. Acute (10 min) anoxia produced a reversible decrease in the sarcolemmal NHE1 activity attributable to NHE1 internalization. Treatment with either 1% O ₂ or dimethyloxaloylglycine (DMOG; 1 mM) for 48-hr stabilized hypoxia-inducible factor 1 and reduced the sarcolemmal NHE1 activity by internalization, but without a change in total NHE1 immunoreactivity or message levels of the coding gene ( SLC9A1) determined in whole-cell lysates. Unlike the effect of DMOG, which was rapidly reversed on washout, reoxygenation after a prolonged period of hypoxia did not reverse the effects on NHE1, unless media were also supplemented with a membrane-permeant derivative of glutathione (GSH). Without a prior hypoxic episode, GSH supplementation had no effect on the NHE1 activity. Thus, posthypoxic NHE1 reinsertion can only take place if cells have a sufficient reservoir of a reducing agent. We propose that oxidative stress under prolonged hypoxia depletes intracellular GSH to an extent that curtails NHE1 reinsertion once the hypoxic stimulus is withdrawn. This effect may be cardioprotective, as rapid postischaemic restoration of the NHE1 activity is known to trigger reperfusion injury by producing an intracellular Na ⁺-overload, which is proarrhythmogenic.     

4-Hydroxynonenal induces membrane perturbations and inhibition of basal prostacyclin production in endothelial cells,and migration of monocytes.

Cultured bovine aortic endothelial cells (BAEC) were incubated for 5 days with 10^?5 4-hydroxynonenal (HN). HN treated BAEC and controls were either (i) further incubated with ¹²⁵I-polymyxin B (IPxB) or with radioiodinated, inactivated coagulation factor Xa (IFXai) as markers of membrane phospholipid perturbation, or (ii) assayed for the synthesis of prostacyclin (PGI₂) and thromboxane A₂ (TXA₂). Rabbit blood mononuclear cells enriched in monocytes (MC) were isolated and assayed for chemotactic response to HN. The results showed six - fold increases of IPxB and IFXai binding to BAEC treated with HN, as compared to untreated controls. We also found in HN treated cells a marked inhibition of PGI₂ synthesis, but an unmodified TXA₂ production. In addition, HN in the 10-5-10-10 M range induced oriented migration of MC.     

Polyphenolic compounds from Geranium pratense and their free radical scavenging activities

Two new polyphenolic compounds, myricetin 3-O-(2"-O-galloyl)-beta-D-glucopyranoside and (-)-6-chloroepicatechin, were isolated from the aerial parts of Geranium pratense subsp. finitimum (Woronow) Knuth, along with three known polyphenolic compounds [quercetin 3-O-(2"-O-galloyl)-beta-D-glucopyranoside, quercetin 3-O-(2"-O-galloyl)-beta-D-galactopyranoside, methyl gallate] and tryptophan. Quercetin 3-O-beta-D-glucopyranoside, quercetin 3-O-beta-D-galactopyranoside, quercetin 3-O-(2"-O-galloyl)-beta-D-glucopyranoside and quercetin 3-O-(2"-O-galloyl)-beta-D-galactopyranoside were found to be effective against free radical induced impairment of endothelium-dependent relaxation in isolated rat aorta.     

Complex compounds of Pd(II), Pt(II), Ni(II), Hg(II) and Cu(I) with 1-benzylidene-2-phenazinoylhydrazine

Synthesis and physical-chemistry characterization of five new genuine complex compounds of 1-Benzylidine-2-phenazinoylhydrazine with Pd(II), Pt(II), Ni(II), Hg(II) and Cu(I) are presented. The chemical structure for these complexes is suggested by the elemental chemical analysis, molecular mass measurements, electric conductivities as well as by IR and UV-VIS spectra. The metal:ligand molar ratio is found 1:1, the obtained complexes belonging to the square planar geometry D4h.     

Luteinizing hormone releasing hormone increases proliferation of meningioma cells in vitro

The fact that meningioma shows at least a 2:1 predilection for women over men is considered to be due to endocrinological and paracrine regulation of the development of this tumour. The presence of receptors for the luteinizing hormone releasing hormone (LHRH) in gynaecological cancer permits the use of LHRH agonistic or antagonistic analogues with a direct effect or by the gonado-pituitary axis suppression in the treatment of these tumours. Therefore, the effect of LHRH on meningioma cells is tested in this study. Meningioma cells from three female patients were cultured and LHRH (50 ng/ml) was added to the growth medium daily, for fourteen days. At the end of this period the cells were counted by means of a Coulter Counter. The stimulating effects of LHRH on the increase of the amount of cells in the meningioma monolayer culture were 146% (p < 0.01), 134% (p < 0.05) and 141% (p < 0.05) of the control, respectively, for the three patients.     

Matrilin-3 is dispensable for mouse skeletal growth and development

Matrilin-3 belongs to the matrilin family of extracellular matrix (ECM) proteins and is primarily expressed in cartilage. Mutations in the gene encoding human matrilin-3 (MATN-3) lead to autosomal dominant skeletal disorders, such as multiple epiphyseal dysplasia (MED), which is characterized by short stature and early-onset osteoarthritis, and bilateral hereditary microepiphyseal dysplasia, a variant form of MED characterized by pain in the hip and knee joints. To assess the function of matrilin-3 during skeletal development, we have generated Matn-3 null mice. Homozygous mutant mice appear normal, are fertile, and show no obvious skeletal malformations. Histological and ultrastructural analyses reveal endochondral bone formation indistinguishable from that of wild-type animals. Northern blot, immunohistochemical, and biochemical analyses indicated no compensatory upregulation of any other member of the matrilin family. Altogether, our findings suggest functional redundancy among matrilins and demonstrate that the phenotypes of MED disorders are not caused by the absence of matrilin-3 in cartilage ECM.     

Enzymatic determination of zinc in vegetables using apocarbonic anhydrase

A new enzymatic method has been developed to determine trace amounts of Zn2+ in vegetables. The basis of the method is that apocarbonic anhydrase regains its activity in proportion to the concentration of Zn2+ present in solution. Bovine carbonic anhydrase was purified from erythrocyte haemolysate by affinity chromatography and the bound Zn2+ removed by dialysis of purified enzyme against a solution of pyridine-2, 6-dicarboxylic acid. Pure (100%) apoenzyme was obtained. The concentration of Zn2+ in vegetable samples was determined using the enzymatic method and by atomic absorption spectroscopy. Determinations made using the two methods were not significantly different one from another.