Biomedical subjects

M Skowronek

Publications and source records attributed to M Skowronek.

11 recordsLinked to original sources

PubMed2003

Selection of biochemical mutants of Aspergillus niger resistant to some abiotic stresses with increased inulinase production.

AIMS: The present work aimed at evaluating the usefulness of selecting different kinds of biochemical mutants of Aspergillus niger to increase inulinase production in submerged culture. METHODS AND RESULTS: Conidia of A. niger 13/36, an active producer of inulinase, were subjected to mutagenesis with both u.v. and N-methyl-N'-nitro-N-nitrosoguanidine (NTG), and the products were analysed for inulinase activity with our own diffusion plate method. As a result of mutagenization and selection for obtaining biochemical mutants (e.g. surviving conditions of certain abiotic stresses, good growing on basal medium at 15 and 40 degrees C), A. niger strains resistant to these agents were obtained. Studies of the relationship between a criterion of selection and the frequency of mutation showed that the highest frequency of positive mutations in the second selection (86%) was obtained in mutants characteristic of the best growth at the low temperature (15 degrees C), when compared with the parent culture (28%). The most active mutants grown under stress conditions showed significantly higher inulinase activity (about 1.2-4.5-fold), when compared with the parent strain. CONCLUSIONS: The studies presented seem to confirm a high effectiveness of selection in some kinds of biochemical mutants of A. niger with regard to increased inulinase activity. SIGNIFICANCE AND IMPACT OF THE STUDY: This screening strategy of mutants can be a contribution to modern commercial enzyme production.

Aspergillus niger↗

PubMed2000

The conformational characteristics of Congo red, Evans blue and Trypan blue.

The structures of the closely related bis-azo dyes Evans blue, Trypan blue and Congo red, which appeared to have different self-assembly properties and correspondingly different abilities to form complexes with amyloids and some other proteins, were compared in this work. Ab initio and semi-empirical methods were used to find the optimal structures and partial charge distributions of the dyes. The optimal structures were searched using different widely used programs. The structures of Congo red and evans blue were found to be planar, except for the torsion on the central diphenyl bond connecting the two halves of the dye. Both symmetrical parts of the molecules appeared very close to planarity. However, Trypan blue exhibits non planarity on the di-azo bonds, as well as on the central bond between the symmetrical parts of the dye. In a consequence, the non planarity of this molecule is higher than in the case of its isomer, Evans blue and Congo red as well. The extra rotation around the azo bonds extorted by the close proximity of the sulfonic groups may be the direct cause of its poor self-assembling and complexation properties versus Evans blue.

Coloring Agents↗

PubMed1998

Supramolecular ligands: monomer structure and protein ligation capability.

The aim of this work was to define the chemical structure of compounds self-assembling in water solutions, which appear to interact with proteins as single ligands with their supramolecular nature preserved. For this purpose the ligation to proteins of bis azo dyes, represented by Congo red and its derivatives with designed structural alterations, were tested. The three parameters which characterize the reactivity of supramolecular material were determined in the same conditions for all studied dyes. These were: A) stability of the assembly products; B) binding to heat-denatured protein (human IgG); and C) binding to native protein (rabbit antibodies in the immune complex) measured by the enhancement of hemagglutination. The structural differences between the Congo red derivatives concerned the symmetry of the molecule and the structure of its non-polar component, which occupies the central part of the dye molecule and is thought to be crucial for self-assembly. Other dyes were also studied for the same purpose: Evans blue and Trypan blue, bis-ANS and ANS, as well as a group of compounds with a structural design unlike that of bis azo dyes. Compounds with rigid elongated symmetric molecules with a large non-polar middle fragment are expected to form a ribbon-like supramolecular organization in assembling. They appeared to have ligation properties related to their self-assembling tendency. The compounds with different structures, not corresponding to bis azo dyes, did not reveal ligation capability, at least in respect to native protein. The conditions of binding to denatured proteins seem less restrictive than the conditions of binding to native molecules. The molten hydrophobic protein interior becomes a new binding area allowing for complexation of even non-assembled molecules.

Congo Red↗

PubMed1997

Effect of self association of bis-ANS and bis-azo dyes on protein binding.

A correlation was found between the ability of dyes (ANS, bis-ANS, Congo red, Evans blue) to form self-associated supramolecular structures in water and their tendency to form complexes with proteins. The self-association ability of dyes was measured as the resistance of a molecular sieve to their penetration. Quantitative evaluation of dye-protein interaction involved measuring the effect of dye on antibodies that agglutinate sheep red blood cells. Enhancement of agglutination by dye was assumed to represent its protein complexation ability. The results confirm that, relative to monomers, self-associated ligands also have altered protein binding properties.

Anilino Naphthalenesulfonates↗

PubMed1997

The use of congo red as a lyotropic liquid crystal to carry stains in a model immunotargeting system--microscopic studies.

The lyotropic liquid crystal dye-Congo Red was used as a carrier in a model immunotargeting system constructed from sheep red blood cells (SRBC) representing the antigen target and rabbit IgG anti-SRBC as the specific driving immunoglobulin. Rhodamine B and Hemin stains were chosen as example chemicals carried to the target. The carried stains were introduced to the micellar organization of Congo Red by intercalation. Preserving its supramolecular organization, Congo Red binds spontaneously and selectively to antibodies that have altered structure extorted by interaction with the antigen in the immune complex. The functionality of the studied immunotargeting model was verified by fluorescence and electron microscopy. The results indicate that the supramolecular nature of protein ligands offers new ligation capabilities possibly useful for carrying stains or drugs in immune-oriented systems.

Animals↗

PubMed1996

Influence of the redox state of ubiquinones and plastoquinones on the order of lipid bilayers studied by fluorescence anisotropy of diphenylhexatriene and trimethylammonium diphenylhexatriene.

The measurements of diphenylhexatriene (DPH) and trimethylammonium diphenylhexatriene (TMA-DPH) fluorescence anisotropy in egg yolk lecithin (EYL) and of DPH anisotropy in dipalmitoylphosphatidylcholine (DPPC) liposomes containing different concentrations of oxidized and reduced ubiquinone (UQ) and plastoquinone (PQ) homologues have been performed. All the oxidized UQ homologues strongly induced ordering of EYL membrane structure, whereas in DPPC liposomes, above the phase transition temperature, the most pronounced effect showed UQ-4. PQ-2 and PQ-9 were less effective than the corresponding ubiquinones in this respect. The reduced forms of UQ and PQ homologues increased the order of membrane lipids to a smaller extent than the corresponding quinones both in the interior of the membrane and closer to its surface. Nevertheless, the investigated prenylquinols showed stronger increase in the membrane order than alpha-tocopherol or alpha-tocopherol acetate, which could be connected with binding of prenylquinol head groups to phospholipid molecules by hydrogen bonds. The strong ordering influence of ubiquinones on the membrane structure was attributed to methoxyl groups of the UQ quinone rings.

Diphenylhexatriene↗

PubMed1996

Location of ubiquinone homologues in liposome membranes studied by fluorescence anisotropy of diphenyl-hexatriene and trimethylammonium-diphenyl-hexatriene.

The measurements of diphenyl-hexatriene (DPH) and trimethylammonium-diphenyl-hexatriene (TMA-DPH) fluorescence anisotropy in dipalmitoylphosphatidylcholine (DPPC) and egg yolk lecithin (EYL) liposomes containing different concentrations of various ubiquinone (UQ) homologues have been performed. UQ-4 induced the highest DPH anisotropy increase in DPPC liposomes, whereas for higher UQ homologues the anisotropy was lowered with the increase of UQ side-chain length. These differences were less pronounced in EYL liposomes. It was concluded that at a higher content in the membranes (3-4 mol%), the short-chain ubiquinones are arranged parallel to lipid fatty acid chains, whereas long-chain homologues are progressively removed from the lipid acyl chains into the midplane region of the membrane. At the lower (1-2 mol%) concentrations, long-chain quinones seem to be evenly distributed within the membrane, especially in EYL membranes. UQ-10 in EYL liposomes perturbed TMA-DPH to a similar extend as the short-chain ubiquinones indicating that UQ-10 penetrates the interface regions of the membrane where its redox reactions occur. The localization and physical state of UQ-10 in native membranes is discussed.

1,2-Dipalmitoylphosphatidylcholine↗

PubMed1996

Congo red-stabilized intermediates in the lambda light chain transition from native to molten state.

Disruption of tertiary interaction makes a protein accessible to penetration by different small molecular compounds. Their interaction may stabilize the altered protein conformation. Congo red is proposed here as a stabilizer of the molten globule state and also of highly reversible intermediates in the transition from native to molten state. Human immunoglobulin lambda light chain (dimer) was used. Two protein-Congo red complexes were found after heating lambda chain in the presence of Congo red. They differed in the amount of attached dye molecules. The binding of dye was interpreted as a two-step dye penetration process involving the peripheral parts of the protein in the first step (at lower temperatures). It was concluded that the liquid crystal properties of Congo red enable it to form specific complexes with proteins, which have become accessible to penetration by ligands after global or local disruption of tertiary interaction. This dye may thus be used as a stabilizer of unfolding intermediates in the step preceding the molten globule state.

Anilino Naphthalenesulfonates↗

PubMed1987

The antiinflammatory action of guanabenz is mediated through 5-lipoxygenase and cyclooxygenase inhibition.

Guanabenz (2,6-dichlorobenzylidene amino guanidine acetate), an alpha 2-agonist, possesses antiinflammatory activity. Since leukotrienes (LT) and prostaglandins (PG) are proinflammatory substances, the effect of guanabenz on LT and PG synthesis by inflammatory cells was investigated. Guanabenz, but not clonidine, B-HT 920 or B-HT 933 inhibited zymosan-induced LTC4 (IC50 = 13 microM) and PGE2 (IC50 = 10.9 microM) synthesis with no concomitant reduction in zymosan phagocytosis or cell viability. Similarly, guanabenz reduced LTB4 (IC50 = 37.4 microM) and PGE2 (IC50 = 13.8 microM) synthesis by A23187-stimulated rat glycogen elicited neutrophils. Furthermore, guanabenz did not inhibit platelet 12-lipoxygenase or phospholipase A2. In vivo, guanabenz was orally active against rat carrageenan paw edema and adjuvant arthritis (ED50s = 9 and 10 mg/kg, respectively). Topically applied guanabenz reduced arachidonic acid (AA)- or tetradecanoyl phorbol acetate (TPA)-induced ear inflammation (ED50s: AA-induced ear edema, 1.4 mg/ear; PMA-induced ear edema, 0.013 mg/ear). Therefore, the antiinflammatory activity of guanabenz may be due to its ability to inhibit the formation of 5-lipoxygenase and cyclooxygenase products.

12-Hydroxy-5,8,10,14-eicosatetraenoic Acid↗

PubMed1986

Synthesis and antiallergic activity of a novel series of 5-lipoxygenase inhibitors.

A series of novel substituted [[(phenoxymethyl)phenyl]amino]oxoalkanoic acid esters have been synthesized. These compounds were tested in vitro for their ability to inhibit the synthesis of 5-hydroxyeicosatetraenoic acid and leukotriene (LT) B4 from rat polymorphonuclear leukocytes (PMN) and in vivo as inhibitors ovalbumin- (OA) and LTD4-induced bronchospasm in the guinea pig. Compounds 5-12 and 25 had IC50's between 1 and 5.6 microM in the rat PMN 5-lipoxygenase assay. Compounds 1, 3, and 16 inhibited OA-induced bronchoconstriction (61%, 64%, and 57%, respectively), but only 1 showed activity against LTD4-induced bronchoconstriction. When tested against LTD4-induced contraction of isolated guinea pig tracheal spiral strips, 1 was a competitive inhibitor with a pKB of 4.94.

Animals↗

PubMed1983

Effects of thiazinamium chloride, promethazine and chlorpromazine on thromboxane B2 synthesis, phagocytosis and respiratory burst by rat alveolar macrophages.

The effects of three phenothiazines, promethazine, thiazinamium chloride and chlorpromazine, on macrophage function were investigated in rat alveolar macrophages. The study focused on thromboxane B2 (TxB2) synthesis, zymosan phagocytosis, and hexosemonophosphate (HMP) shunt activity in these phagocytes. TxB2 synthesis by resting macrophages was inhibited by thiazinamium chloride and promethazine in a dose-dependent manner. However, chlorpromazine was inhibitory only at 10(-3) M. Promethazine treatment of zymosan-activated macrophages led to a concomitant reduction in both phagocytosis and TxB2 synthesis. Thiazinamium chloride inhibited TxB2 synthesis but had no effect on the ingestion of zymosan particles. In contrast, chlorpromazine inhibited phagocytosis but not TxB2 synthesis except at 10(-3) M. The effects of these agents on the formation of TxB2 synthesis from exogenous arachidonic acid were also investigated. Under these conditions where indomethacin, a known cyclooxygenase inhibitor, was inhibitory, promethazine but not thiazinamium chloride inhibited TxB2 synthesis from exogenous arachidonic acid. Treatment of macrophages with promethazine and chlorpromazine but not thiazinamium chloride results in a reduction in the oxidative burst during phagocytosis. The results suggest that the phenothiazines used in this study differ from one another in their actions on macrophage function. Furthermore, the ability of thiazinamium chloride to selectively inhibit arachidonic acid metabolism may contribute to its bronchodilator/antiallergic activity.

Animals↗