Showing papers on "Glutaraldehyde published in 1979"
TL;DR: Conditions for the gelation of k ‐carrageenan, which is a new polymer for immobilization of enzymes and microbial cells, were investigated in detail and immobilized preparations were easily tailor‐made to various shapes such as cube, bead, and membrane.
Abstract: Conditions for the gelation k-carrageenan, which is a new polymer for immobilization of enzymes and microbial cells, were investigated in detail k-Carrageenan was easily induced to gel by contact with metal ions, amines, amino acid derivatives, and water-miscible organic solvents By using this property of k-carrageenan, the immobilization of enzymes and microbial cells was investigated Several kinds of enzymes and microbial cells were easily immobilized with high enzyme activities Immobilized preparations were easily tailor-made to various shape such as cube, bead, and membrane The obtained immobilized preparations were stable, and columns packed with them were used for continuous enzyme reaction for a long period Their operational stabilities were enhanced by hardening with glutaraldehyde and hexamethylenediamine
154 citations
TL;DR: It is concluded that at physiological pH: most of the charged proteins and lipids on cell surface can contribute to the membrane surface charge, and the membranes surface charge of cells can be greatly changed by chemical fixation.
Abstract: The surface charge of cultured neurons was investigated with the electron microscope markers anionized ferritin (AF) and cationized ferritin (CF). To determine which membrane components could react with the markers, model reactions were used. Both protein-coated Sepharose beads and lipid vesicles were reacted at physiological pH. Results with these model reactions indicate that the following groups may contribute to the surface charge: acidic groups--the sialic acid of both glycoproteins and gangliosides, the carboxyl group of proteins, and the phosphates of phospholipids; basic groups--the amines of proteins. The effect of chemical fixation on the surface charge was investigated. Glutaraldehyde fixation was shown to increase the charge of neutral proteins but not by a mechanism involving unbound aldehydes. Glutaraldehyde fixation of phospholipid vesicles in the presence of CF showed that amine-containing phospholipids were cross-linked to CF. This cross-linkage was seen with the electron microscope as the clumping of CF and the burying of CF in the membrane. Paraformaldehyde fixation had a lesser effect on the charge of proteins but did react with phospholipids as did glutaraldehyde. It is concluded that at physiological pH: (a) most of the charged proteins and lipids on cell surface can contribute to the membrane surface charge, and (b) the membrane surface charge of cells can be greatly changed by chemical fixation.
93 citations
TL;DR: The observations indicate that there are several options for the localization of catalase depending on the fixation and incubation conditions, and these conditions should be selected according to the tissue and the purpose of the study.
Abstract: The influence of various parameters of fixation and incubation upon the oxidation of DAB by catalase have been analyzed. Crystalline beef liver catalase was fixed with different concentrations of glutaraldehyde and peroxidatic activity was determined spectrophotometrically using DAB as hydrogen donor. Although aldehyde fixation appeared to be important in elicitation of the peroxidatic activity of catalase, the final pigment production after 60 min incubation was optimal with the lowest concentration of glutaraldehyde (1%), after the shortest fixation period (30 min), and at the lowest temperature (5° C) tested. Similarly cytochemical studies with rat kidney sections incubated for 10 min confirmed that the staining of peroxisomes in proximal tubules was strongest after the “mildest” fixation conditions. The pH and the temperature of incubation were closely interrelated, so that at room temperature (25° C) the maximal pigment production was obtained at pH 10.5 but incubation at 45° C gave the strongest staining at pH 8.5. The production of pigment increased with higher DAB concentrations which required larger amounts of H2O2 in the incubation medium. Cytochemical studies on renal peroxisomes were in agreement with these biochemical findings. The observations indicate that there are several options for the localization of catalase depending on the fixation and incubation conditions. Hence, these conditions should be selected according to the tissue and the purpose of the study. Examples for such selective applications are presented.
75 citations
TL;DR: E. coli cells immobilized with к-carrageenan and treated with glutaraldehyde and hexamethylenediamine showed the highest productivity, and was compared for production of l - aspartic acid.
72 citations
Journal Article•
TL;DR: A simplified technique to prepare fungal specimens for scanning electron microscopy is described and discussed in this paper, where the minimum fixation time in OSO4 was 2 h, in glutaraldehyde 4 to 5 h.
Abstract: A simplified technique to prepare fungal specimens for scanning electron microscopy is described and discussed. Fixation in either 6% glutaraldehyde or 2% Os04 (both unbuffered aqueous solutions), yielded good results. The minimum fixation time in OSO4 was 2 h, in glutaraldehyde 4 to 5 h. Chemical dehydration with 2-methoxyethanol or 2,2-dimethoxypropane proved to be useful since it considerably reduced the preparation time. Because of the relatively few changes of reagent solutions during the whole process, the fungal specimens were less disturbed without much loss of material. The preparation technique described has been applied to specimens of various fungal groups with good results.
60 citations
TL;DR: It is concluded that the strong chromophore shown by glutaraldehyde-treated proteins is due to the formation of such quaternary pyridinium compounds and that anabilysine residues are important in the cross-linking of proteins by glutARaldehyde.
Abstract: 1-(5-Amino-5-carboxypentyl)pyridinium chloride (3) and 1-(5-amino-5-carboxypentyl)-3-[1-(5-amino-5-carboxypentyl)-2-piperidyl]pyridinium chloride, anabilysine (4), have been isolated from acid hydrolysates of glutaraldehyde-treated ovalbumin and their structures confirmed by synthesis. It is concluded that the strong chromophore at ca. 265 nm shown by glutaraldehyde-treated proteins is due to the formation of such quaternary pyridinium compounds and that anabilysine residues are important in the cross-linking of proteins by glutaraldehyde.
56 citations
TL;DR: A reliable Golgi-Kopsch variant is described and procedures for cutting, counterstaining, and handling sections are described.
53 citations
TL;DR: Based upon its superior catalytic activity for H2O2 decomposition, a bituminous coal‐based activated carbon was selected for investigations of pretreatment and enzyme immobilization methods.
Abstract: Based upon its superior catalytic activity for H2O2 decomposition, a bituminous coal-based activated carbon was selected for investigations of pretreatment and enzyme immobilization methods. Pretreatments considered include acid washing, exposure to strong oxidizing agents, contact with concentrated peroxide solutions, nitration and amination, isothiocyanate derivatization, silanization, and stearic acid coating. Effects of these pretreatments on morphology and trace-metal content of the carbon pellets have been studied using scanning electron microscopy and dispersive analysis of x rays. Immobilization of glucoamylase by adsorption, glutaraldehyde crosslinking, and covalent attachment to carbon activated by water-soluble diimide or diazotization have been examined. These different enzyme-carbon catalysts have been characterized by their enzyme loading, enzyme activity, catalytic activity for H2O2 decomposition, or combinations of these measures of performance.
48 citations
TL;DR: The enzyme (urease amidohydrolase, EC 3.5.1.5) prepared from Cajanus indicus, has been immobilized with glutaraldehyde‐treated chitin as the solid support and the storage stability of the chit in–urease system was determined.
Abstract: The enzyme urease (urea amidohydrolase, EC 3.5.1.5) prepared from Cajanus indicus, has been immobilized with glutaraldehyde-treated chitin as the solid support. The immobilized enzyme was characterized by determining the pH profiles and optimum temperature. Effect of glutaraldehyde concentration on the binding of enzyme to chitin was studied. The storage stability of the chitin-urease system was determined.
32 citations
01 Mar 1979
TL;DR: Lettuce thylakoids were immobilized by the action of glutaraldehyde at sub-zero temperature in presence of albumin and particles with sponge structure and good mechanical properties were obtained.
Abstract: Lettuce thylakoids were immobilized by the action of glutaraldehyde at sub-zero temperature in presence of albumin. Particles with sponge structure and good mechanical properties were obtained. The activity yield after immobilization was found equal to 70% for the oxygen production. Photosystems I and II were both active after immobilization. The yield for the ATP production was 27%. An increase of stability of the immobilized thylakoids was observed when stored under illumination.
31 citations
TL;DR: Inhibition of the enzyme-linked immunosorbent assay was used as a sensitive technique for measuring antigenic activity of antigen E or its derivatives.
TL;DR: In this article, the magnesium salt of sulphated lauryl alcohol was shown to potentiate and stabilize the biocidal activity of acid glutaraldehyde, and the effect of operating temperature and storage temperature on sporicidal activity was also examined.
TL;DR: Rat brains were fixed either with glutaraldehyde (GA) or formaldehyde (FA) and Ultrastructural preservation was good after only 20 min GA-fixation, provided that the subsequent treatment was with isotonic solutions.
Abstract: Rat brains were fixed either with glutaraldehyde (GA) or formaldehyde (FA). After 20 min or 24 h fixation the osmotic sensitivity of the tissue was tested by immersion in (a) distilled water (b) 0.15 M or (c) 0.3 M cacodylate buffer. GA-fixed material retained some sensitivity to osmotic stress after 20 min fixation but was entirely resistant after 24 h fixation. Ultrastructural preservation was good after only 20 min GA-fixation, provided that the subsequent treatment was with isotonic solutions. The fixation with FA was less efficient and slower. Dark neurons and other artifacts were commonly seen after the 20 min fixation with FA. Prolongation of the FA-fixation overnight gave markedly better preservation, but however, never equivalent to that with GA.
TL;DR: The inability of glutaraldehyde to crosslink low concentrations of protein has consequences for the interpretation of the intracellular ultrastructural location of virus or virus-related products in expanded leaf tissues as discussed by the authors.
Patent•
11 Apr 1979
TL;DR: In this article, a branched polyethylene imine is used to immobilize intracellular glutaraldehyde sensitive enzymes in the presence of a polyamine which is preferably a polyine.
Abstract: Intracellular glutaraldehyde sensitive enzymes are immobilized by reacting microbial cell material with glutaraldehyde in the presence of a polyamine which is preferably a branched polyethylene imine.
TL;DR: An electron microscopic study was carried out to examine the quality of ultra‐structural preservation of parenchymatous and mesenchym atous tissues and isolated cells fixed in glutaraldehyde with sodium azide (NaN3) as an additive.
Abstract: An electron microscopic study was carried out to examine the quality of ultrastructural preservation of parenchymatous and mesenchymatous tissues and isolated cells fixed in glutaraldehyde with sodium azide (NaN3) as an additive. The dense tissues fixed with conventional glutaraldehyde containing calcium chloride demonstrated only a narrow zone of good tissue preservation on the surface of the specimens. Addition of azide at a concentration of 0.1% greatly improved the cellular preservation in the deeper region of tissues, in particular with respect to the mitochondrial morphology. There was no adverse effect on other cell organelles. The improvement in mitochondrial preservation and the enhancement of penetration of the fixative is presumably due to selective and instantaneous inhibition of mitochondrial metabolic activity by the azide, thus retarding anoxic degenerative effects on cellular structures until permanent fixation is completed by the comparatively slow-acting aldehyde. However, the addition of azide offers no significant improvement in the ultrastructural preservation of isolated lymphocytes and liver cells, or fibroblasts maintained in culture.
TL;DR: The antigenic identity of the native enzyme appeared to be lost in the glutaraldehyde derivative, although it was not altered by an analogous modification with a monofunctional reagent, and the Michaelis constants of the enzyme were not affected by cross-linking.
TL;DR: In the present study the conditions leading to tight binding of human serum albumin to the Amberlite XAD-7 resin without the use of chemical coupling agents have been defined andoretical analysis of the data suggest adsorption of a monolayer of albumin which is supported by the absence of visual surface coating on scanning electron micrographs.
Abstract: In the present study the conditions leading to tight binding of human serum albumin to the Amberlite XAD-7 resin without the use of chemical coupling agents have been defined. Optimal binding (10.97 mm/kg dry XAD-7) was achieved at pH 5.0 and adsorption conformed to a Langmuir isotherm. Theoretical analysis of the data suggest adsorption of a monolayer of albumin which is supported by the absence of visual surface coating on scanning electron micrographs. Binding of human serum albumin was reduced when two chemical coupling agents, glutaraldehyde and carbodiimide were included in the human serum albumin solution, the elution of adsorbed HSA from the resins under severe flow conditions was reduced, and the amount eluted was minimal in all instances.
TL;DR: Trypsin, pronase and subtilisin were immobilized on chitosan by glutaraldehyde coupling and found to be less sensitive to its natural inhibitors than the soluble enzyme.
Abstract: Trypsin, pronase and subtilisin were immobilized on chitosan by glutaraldehyde coupling. Significant retention of activity was observed when synthetic substrates as well as casein were used. The specific activities of the bound proteinases ranged from 38% to 79% of their initial specific activities. The pH-activity profile of trypsin was slightly shifted toward alkaline values, and its thermal stability was increased. Immobilized trypsin was found to be less sensitive to its natural inhibitors than the soluble enzyme.
TL;DR: This study attempted to use paraformaldehyde-glutaraldehyde modified after Karnovsky (1965) for routine light and electron microscopic study of phytoflagellates to avoid toxicity problems and provide samples that would be useful for light microscopy, SEM, and TEM.
TL;DR: Reconstituted thin filaments of chicken gizzard were cross-linked with glutaraldehyde either in the presence or absence of Ca2+.
Abstract: Reconstituted thin filaments (the actin-tropomyosin-leiotonin complex) of chicken gizzard were cross-linked with glutaraldehyde either in the presence or absence of Ca2+. The ability of resultant thin filaments to activate myosin ATPase was 'frozen' in the activated or inactivated state, respectively. This result clearly indicates the existence of actin-linked regulation in smooth muscle.
TL;DR: Cells of Arthrobacter X-4 were immobilized by entrapment in gelatin crosslinked with glutaraldehyde and xanthine oxidase activity and stability were determined at various temperatures.
Abstract: Cells of Arthrobacter X-4 were immobilized by entrapment in gelatin crosslinked with glutaraldehyde. The xanthine oxidase activity and stability were determined at various temperatures. In comparison with bovine milk xanthine oxidase the bacterial enzyme is more stable and has a different substrate specificity. 1-Methylxanthine was oxidized on a preparative scale.
Journal Article•
TL;DR: Polyhydroxybutyrate granules, generally poorly preserved in thin sections of Aquaspirillum serpens, were well preserved by the malachite green-glutaraldehyde fixative.
Abstract: Malachite green combined with glutaraldehyde has been used recently as a fixative for preserving and revealing lipid complexes in thin sections of eukaryotic cells examined by electron microscopy. When bacteria were prefixed with the above mixture granular electron dense inclusions were revealed in all cultures tested. These inclusions were replaced by electron transparent areas in cells fixed with glutaraldehyde alone. The structures were frequently located near to or within the nucleoid and adjacent to the cell membrane in Gram-negative bacteria and were associated with the nucleoid and mesosomes in Gram-positive species. Polyhydroxybutyrate granules, generally poorly preserved in thin sections of Aquaspirillum serpens, were well preserved by the malachite green-glutaraldehyde fixative. Malachite green complexes were observed outside of the cells in all preparations. Capsules were neither preserved nor stained.
TL;DR: The results indicate that formaldehyde is the component of formocresol that interacts with the protein portion of bovine pulp tissue and that Glutaraldehyde mimics many of the fixative properties of formaldehyde and improves on others, thereby justifying future testing of glutaraldehyde as a pulpotomy agent.
TL;DR: The present findings establish that although the expression of a tumour-specific antigen is necessary for the induction of immuno-protection against tumOUR-cell challenge, this alone is not a sufficient condition for eliciting tumour immunity.
Abstract: gamma-Irradiated rat hepatoma cells are immunogenic in syngeneic WAB/Not rats, so that immunized animals are protected against tumour-cell challenge and circulating tumour-specific antibody is produced. Treatment of the immunizing cells with glutaraldehyde at concentrations of 0.001% or greater for 30 min rendered these cells non-protective in tumour-rejection tests and no longer able to induce significant formation of specific antibody. However, tumour-specific antigens were shown to be expressed upon treated cells; they specifically bound tumour-specific antibody from syngeneic immune sera assessed in indirect membrane-immunofluorescence tests. Also, these cells specifically absorbed antibody from immune or tumour-bearer sera, as demonstrated in the indirect membrane-immunofluorescence test or a complement-dependent 51Cr-release test. Alloantigen expression was not influenced by glutaraldehyde treatment, although glutaraldehyde-treated hepatoma cells failed to induce alloantibody formation in KX/Not rats. Polyacrylamide-gel electrophoresis of treated cells, surface-labelled with 125I, indicated that extensive cross-linking of the surface protein occurred as a result of glutaraldehyde treatment. The present findings establish that although the expression of a tumour-specific antigen is necessary for the induction of immuno-protection against tumour-cell challenge, this alone is not a sufficient condition for eliciting tumour immunity.
TL;DR: The most remarkable observation of the study was that the densitometric method yielded reproducible results and that the different fixatives gave significantly different values for the relative thickness of cellular membranes.
Abstract: A densitometric method was utilized in the measurement of the relative thickness of the cellular membranes in the ventral lobe of the rat prostate. Potassium permanganate, glutaraldehyde, osmium tetroxide, and ruthenium tetroxide solutions were used as fixatives. During preparation for electron microscopy, the tissues were given standardized treatments to reduce methodological errors; latex particles were applied to the thin sections to serve as reference particles of a known size. The most remarkable observation of the study was that the densitometric method yielded reproducible results and that the different fixatives gave significantly different values for the relative thickness of cellular membranes. Glutaraldehyde, or glutaraldehyde followed by ruthenium tetroxide post-fixation, gave the highest values for membrane thickness while osmium tetroxide and potassium permanganate gave the lowest values. Glutaraldehyde treatment, prior to osmium tetroxide or potassium permanganate post-fixations, rendered the membranes thicker than after osmium tetroxide and potassium permanganate treatments alone. Ruthenium tetroxide appeared to be very suitable for fixation of cellular membranes.
TL;DR: The recovery of N-acetyl-beta-D-hexosaminidase after immobilization was unaffected by increasing the concentration of glutaraldehyde, but was decreased by increasingThe bovine serum albumin concentration, and the immobilized enzyme showed enhanced resistance towards proteolytic and thermal inactivation.
TL;DR: Increased bactericidal activity was observed on addition of inorganic cations to both acid and alkaline glutaraldehyde solutions and Mg2+ ion in particular potentiated activity and this was attributed to cell surface effects due to osmotic forces.
Abstract: Increased bactericidal activity was observed on addition of inorganic cations to both acid and alkaline glutaraldehyde solutions. Mg2+ ion in particular potentiated activity and this was attributed to cell surface effects due to osmotic forces resulting in greater uptake of disinfectant, reduction in cell volume and interference with enzyme activity associated with the cell wall.
01 Mar 1979
TL;DR: Results suggest that covalent attachment via spacer is strongly influenced by cell surface characteristics.
Abstract: The potential of hydroxyalkyl methacrylate gel as a support for yeast cell immobilization by covalent attachment was investigated with respect to the length of spacer used and the manner of its activation. Saccharomyces paradoxus cells were immobilized successfully onto glutaraldehyde activated glycyl-, β-alanyl-and e-aminocaproyl-derivatives of the above mentioned gel modified with hexamethylenediamine. Results suggest that covalent attachment via spacer is strongly influenced by cell surface characteristics.