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PubMed1989

[Ultrastructural morphometric studies of myocardial cells of rats after 3 hour autolysis with and without preliminary short term physical exertion].

The influence of a period of 3 h of autolysis on the ultrastructure of the heart muscle cells of 2 groups of rats was examined. One group was undergone 5 h of swimming training and the other remains without such training. The results indicate that some ultrastructural changes caused by the exercise are the same even after 3 h of autolysis. On the other hand, swimming training before autolysis affects the process of autolysis.

Animals↗

PubMedDate not supplied

[Changes in phospholipids in the gray and white matter of the rat brain during postmortem autolysis dynamics].

Content of total phospholipids and their individual fractions, studied in rat brain grey and white substances during postmortal autolysis, was distinctly and dissimilarly altered in these tissues and the rate of alterations depended on time of incubation. Relative content of phosphatidyl serines + sphingomyelins, phosphatidic acids + polyglycerophosphatides in grey substance as well as of phosphatidylinositides, phosphatidic acids + polyglycerophosphatides in white substance were increased, while phosphatidylinosidtides in grey substance and phosphatidyl serines + sphingomyelins in white substance were decreased within later (4 and 24 hrs) periods of autolysis. Content of phosphatidyl cholines, phosphatidyl ethanolamines, glycerophosphates and lysophospholipids was also dissimilarly altered in brain grey and white substances both within early and later periods of autolysis. The schemes of metabolic interrelations between phospholipids of rat brain grey and white substances during autolysis are considered involving hydrolase, transferase and other reactions of lipids biotransformation. This multiple complex of biochemical reactions may be among the reasons responsible for dissimilar and time-dependent deteriorations of individual brain structures after death.

Animals↗

PubMedDate not supplied

[Synaptic membrane lipids from rat brain during autolysis].

Content of total lipids, phospholipids and total protein was decreased in rat brain synaptic membranes during autolysis within 1 h and 24 hrs at 37 degrees in the medium used for isolation of the membranes. Within early steps of autolysis (1 h), total lipids and phospholipids were only slightly altered, while a decrease in the concentrations of phosphatidyl cholines, phosphatidyl ethanolamines and cholesterol was found simultaneously increased in relation to the content of lysophospholipids, phosphatidyl serines+sphingomyelins, cholesterol esters, triacylglycerols and free fatty acids. Within later periods of autolysis (24 hrs) the further dissimilar decrease was observed in relative content of phosphatidyl cholines, phosphatidyl ethanolamines, phosphatidyl serines+sphingomyelins with synchronous increase in the level of glycerophosphates, lysophospholipids, phosphatidic acid+polyglycerophosphatides, cholesterol esters; occurrence of diacylglycerols was detected. Possible mechanisms (involving hydrolase, transacylase reactions) of alterations observed in lipids from synaptic membranes during autolysis are discussed. Modern experimental technique appears to be useful in order to gain insight into molecular mechanisms of brain death and the postmortal loss of information.

Animals↗

PubMedDate not supplied

[Lipids from gray and white rat brain matter in autolysis].

A decrease in relative content of phospholipids and cholesterol simultaneously with increase in cholesterol esters and free fatty acids were detected in tissues of rat brain gray and white matters during autolysis at 37 degrees within 6-7 min, 1, 4 and 24 hrs; the most distinct alterations were observed in lipids of the gray matter especially at early (6-7 min) and late (24 hrs) stages of autolysis. In the gray matter, relative content of all the lipid fractions studied was restored to initial level within 4 hrs of incubation. In the white matter, during autolysis the content of cholesterol varied, the content of phospholipids was only slightly increased, while the level of free fatty acids was increased only at 24 hrs of incubation with synchronous decrease in content of triacylglycerols, cholesterol and, partially, phospholipids. These experimental data are of importance in resuscitation. Both common and dissimilar mechanisms of these lipid alterations are discussed. Not only the complex of hydrolase reactions but also that of transacylase and other reactions, involved in the degradation and biotransformation of brain lipids in autolysis, were noted.

Animals↗

PubMed1997

Densitometric and chromatin texture parameters of hepatocyte nuclei during controlled autolysis.

OBJECTIVE: To determine the extent of the effect of autolytic changes in hepatocyte nuclei on the results of measurements of integrated optical density (IOD) of nuclei stained by the Feulgen method and its possible relationship with changes in chromatin texture. STUDY DESIGN: Cytologic smears obtained from five pig livers by means of fine needle biopsy were fixed in 10% formalin at six time points after slaughtering (0-18 hours) and stained per Feulgen. In the hepatocyte nuclei IOD measurements were performed and supplemented with quantitative texture analysis. Texture analysis was carried out with our own algorithm, based on the movement of global gravity centers of heterochromatin granules during two-level, iteratively changed, grey thresholding. RESULTS: The variability coefficients of IOD of diploid hepatocytes during autolysis increased from 0.061 (0 hours) to 0.363 (18 hours), and the ploidy histograms became aneuploid. The texture of chromatin became more homogeneous, but other features of the texture did not indicate a significant correlation with the duration of autolysis. CONCLUSION: Autolysis is an important factor, which may influence the accuracy of DNA content and shape of ploidy diagrams. Changes in the morphologic pattern of heterochromatin may play a role in this, but the study was not conclusive because only one of six texture parameters (number of granules) changed parallel to mean IOD and autolysis time.

Animals↗

PubMed1976

[Influence of autolysis on the quantitative cytoarchitecture of rat hepatocytes. (An ultrastructural morphometric study) (author's transl)].

INTRODUCTION: Autolysis is very often a reason for cell damage and is also super-imposed on many other cell damages. Already one hour of autolysis causes serious changes in cell metabolism, which can be demonstrated morphometrically. MATERIAL AND METHODS: Experiments were made with 10 male adult Wistar rats. 5 animals were for control purposes and 5 animals had to undergo a 1-hour's autolysis. The morphometric analysis of the liver parenchymal cells was based on the information of Weibel et al. (1968) with the help of a computer program. RESULTS AND DISCUSSION: The consequence of the drop in energy caused by autolysis is an enlargement of the liver cell which is possibly due to a breakdown of the energy-dependent ionic pumps. One of the earliest observable cell changes is a so-called "Kernwandhyperchromatosis" and chromatin condensation within the nuclei, whereby the Kernwandhyperchromatosis is seen to be a direct consequence of the increase in lactate and decrease of pH. ATP-sufficiency causes a disturbed function of mitochondrial membranes. The microchondria are swollen, the number of mitochondrial grana is clearly reduced. An enlargement of the mitochondrial outer membrane takes place by folding while the surface of mitochondrial cristae remains unchanged. As a consequence of the altered membrane activities also the peroxisomes swell at reduced numerial density. At unchanged total volume of RER the surface of the granulated membranes of the RER decrease by 50%. This decrease caused by ribosome detachment of the granulated membranes corresponds to the enlargement of the degranulated membrane parts of the nedoplasmic reticulum. The vesiculation is caused by an unspecific damage of cytoplasm. While the density of its volume and the membrane surface remain unaltered, the SER also shows a tendency to small vesiculation caused by an unspecific damage of cytoplasm. The increase in number and volume of the lysosomes and vacuoles of unknown origin speaks for a lysosomal activity. The cell compartment responsible for protein synthesis shows the most impressive morphometric and morphologic changes, which eventually can be explained by a decrease of protein synthesis which is needed to obtain enough energy for a well operating physiological equilibrium.

Animals↗

PubMed1993

Recognition and reduction of artifacts from autolysis in paraffin-embedded tissue using DNA/nuclear protein flow cytometry.

Artifacts from autolysis can be a problem in retrospective flow-cytometric analyses of DNA content in paraffin-embedded tissues. Autolyzed tissue from rat liver, human liver, and rat spleen were stained for DNA and nuclear protein to determine if this technique would be useful in identifying partially degraded cells. After the tissue was deparaffinized and rehydrated, the nuclei were isolated using 0.5% pepsin. Propidium iodide (PI) and fluorescein isothiocyanate (FITC) were used to stain DNA and nuclear protein. When unfixed rat liver tissue was allowed to undergo autolysis at 4 degrees C for 24-48 h before fixation, there was a progressive broadening of the G1 and G2M DNA peaks and a slight increase in the average DNA contents of these peaks. Nuclei that stained more intensely with PI also stained more intensely with FITC. Similar results were obtained using human liver and rat spleen. Sometimes the increased PI staining resulted in a false aneuploid peak. The distinctive skewing of the DNA/nuclear protein histograms from autolysis was reduced by increasing the incubation of the tissue in 0.5% pepsin from 0.5 h to 1.5 h during the nuclei-isolation step. The DNA/nuclear protein method provides a means for identifying artifacts from autolysis, whereas the extended pepsin treatment provides a means for reducing these artifacts.

Animals↗

PubMed1979

[Course and modification of Hypomyces ochraceus m 359 autolysis].

Growth and natural autolysis of the ascomycete Hypomyces ochraceus were investigated in shake cultures by measuring dry weight, pH value, and uptake of carbohydrate. The alkaline autolysis resulted in maximum loss of 53% of dry weight during an incubation period of 35 days. Early autolysis was induced by limitation of oxygen, increasing temperature, addition of peracetic acid, and sodiumacide. Autolysis is discussed to be derived from deficiency of energy.

Azides↗

PubMed1976

Lytic enzymes in the autolysis of filamentous fungi.

The degrees of autolysis attained by five different genera of filamentous fungi during an incubation period of 60 days, under the same culture conditions were: 87.3% for Penicillium oxalicum; 65.9% for Neurospora crassa; 62.7% for Polystictus versicolor; 51.7% for Aspergillus niger and 23.5% for Nectria galligena. N. crassa, A. niger and P. versicolor reached the end of the autolysis during this incubation period (60 days), whereas P. oxalicum and N. galligena did not. The excretion of the lytic enzymes beta-N-acetylglucosaminidase, beta -1-3 glucanase, chitinase, invertase and acid phosphatase into the culture medium during growth and autolysis was investigated. The excretion of these enzymes was consistent with the degree of autolysis reached, the maximum excretion belonging to P. oxalicum and the minimum to N. galligena. The N. crassa invertase was excreted into the culture liquid at levels very much higher than the other enzymes studied, and at levels very much higher than the invertases excreted by the other fungi.

Acetylglucosaminidase↗

PubMed1980

Glucose repression and autolysis of Saccharomyces cerevisiae cells: alterations in the cytochemical localization of acid phosphatase.

Alterations in the localization of acid phosphatase in Saccharomyces cerevisiae during glucose repression and during autolysis have been studied. Cell morphology becomes distinctly changed after only 2 h in the presence of high glucose concentration while after 3 h of glucose repression the majority of the mitochondrial structures resemble promitochondria. Yeast cells repressed for 6 h contain almost completely degraded mitochondrial structures and numerous lipid droplets in the central vacuole and cytoplasm. Destruction of mitochondria is accompanied by the accumulation of acid phosphatase in these organelles and in the cytoplasm, whereas its activity in the central vacuole is lowered, most probably because of the leakage of the enzyme into the cytoplasm. No preferential breakdown of mitochondria is observed during autolysis. On the contrary, mitochondria are apparently the last to be degraded. Digestion of cytoplasmic regions and membranous elements occurs intravacuolarly after sequestration by protrusions of the central vacuole which are formed at the initial stages of autolysis. Acid phosphatase is not released from the central vacuole, suggesting indirectly that vacuole enzymes do not migrate into the cytoplasm during autolysis.

Acid Phosphatase↗

PubMed1999

Autolysis of Escherichia coli and Bacillus subtilis cells in low gravity.

The role of gravity in the autolysis of Bacillus subtilis and Escherichia coli was studied by growing cells on Earth and in microgravity on Space Station Mir. Autolysis analysis was completed by examining the death phase or exponential decay of cells for approximately 4 months following the stationary phase. Consistent with published findings, the stationary-phase cell population was 170% and 90% higher in flight B. subtilis and E. coli cultures, respectively, than in ground cultures. Although both flight autolysis curves began at higher cell densities than control curves, the rate of autolysis in flight cultures was identical to that of their respective ground control rates.

Bacillus subtilis↗

PubMed2005

Enhanced production of manganese peroxidase from immobilized Phanerochaete chrysosporium due to the increased autolysis of chlamydospore-like cells.

The autolysis of chlamydospore-like cells in Phanerochaete chrysosporium immobilized in polyurethane foam correlated with the production of manganese peroxidase (MnP). The maximum specific activity of MnP was 1055 U g dry mycelium(-1) in the immobilized culture, compared with 260 U g dry mycelium(-1) in the submerged culture. Scattered mycelial pellets were formed in the immobilized culture in which almost all of the chlamydospore-like cells were subject to autolysis. However, highly crowded pellets were formed in the free culture, in which only the chlamydospore-like cells in the exterior were subject to autolysis. We propose that the enhanced production of MnP in immobilized cultures of P. chrysosporium is due to increased autolysis of the chlamydospore-like cells.

Biomass↗

PubMed1986

Transcription but not translation is required for EDTA-induced autolysis in Escherichia coli.

Rifampicin, but not chloramphenicol or other inhibitors of translation, inhibited EDTA-induced autolysis in Escherichia coli. Inhibition of EDTA-induced autolysis in E. coli was also observed with nalidixic acid and novobiocin, inhibitors of topoisomerase II. Rifampicin or nalidixic acid-resistant mutants of E. coli were resistant to the inhibitory effect of the respective antibiotic on EDTA-induced autolysis. The implications of these studies in regard to our understanding of the regulation of autolysis in E. coli are discussed.

Bacteriolysis↗

PubMedDate not supplied

Evidence for yeast autophagy during simulation of sparkling wine aging: a reappraisal of the mechanism of yeast autolysis in wine.

Yeast autolysis is the source of several molecules responsible for the quality of wines aged in contact with yeast cells. However, the mechanisms of yeast autolysis during wine aging are not completely understood. All descriptions of yeast autolysis in enological conditions emphasize the disturbance of cell organization as the starting event in the internal digestion of the cell, while no reference to autophagy is found in wine-related literature. By using yeast mutants defective in the autophagic or the Cvt pathways we have demonstrated that autophagy does take place in wine production conditions. This finding has implications for the genetic improvement of yeasts for accelerated autolysis.

Autophagy↗

PubMed1987

The effects of autolysis on the structure of chicken calpain II.

When chicken calpain II autolysed in the presence of Ca2+, it underwent limited proteolysis to give peptides of Mr 54,000 and 37,000, and several of Mr approx. 30,000 and 18,000. The autolytic peptides were purified and their N-terminal amino acid sequences determined. By comparison of these sequences with the known sequence of the complete calpain molecule, the autolytic cleavage sites were identified. The structural integrity of the molecule during autolysis was investigated by gel-permeation chromatography. Experiments were also done to test the reversibility of adding EDTA to calpain during autolysis, measured as recoverable enzyme activity assayed in the presence of Ca2+. The results are presented in terms of a model for the structural changes occurring in calpain during autolysis. It was concluded that the loss of enzymic activity, which is a consequence of autolysis, was due to dissociation of the autolytic peptides after cleavage of the calpain large subunit within the third domain.

Amino Acid Sequence↗

PubMed1997

Oxidation inhibits substrate proteolysis by calpain I but not autolysis.

In this study, the effects of oxidation on calpain I autolysis and calpain-mediated proteolysis were examined. Calpain I was incubated with increasing concentrations of free calcium in the presence or absence of oxidant, and autolytic conversion of both the 80- and 30-kDa subunits was measured by immunoblotting utilizing monoclonal antibodies which recognize both autolyzed and non-autolyzed forms of each subunit, respectively. Autolytic conversion of the 80-kDa subunit of calpain I was not detected until free calcium concentration was greater than 40 microM, whereas autolysis of the 30-kDa subunit did not occur until the free calcium concentration was greater than 100 microM. In addition, autolytic conversion of either the 80- or 30-kDa subunit was not inhibited by the presence of oxidant. Calpain I activity was measured using the fluorescent peptide N-succinyl-L-leucyl-L-leucyl-L-valyl-L-tyrosine-7-amido-4- methylcoumarin or the microtubule-associated protein tau as substrate. Calpain I was found to have proteolytic activity at free calcium concentrations below that required for autolysis. Calpain I activity was strongly inhibited by oxidant at all calcium concentrations studied, suggesting that proteolytic activity of both the non-autolyzed 80-kDa and autolyzed 76-kDa forms was susceptible to oxidation. Interestingly, whereas oxidation did not inhibit autolytic conversion, the presence of high substrate concentrations did result in a significant reduction of autolysis without altering calpain proteolytic activity. Calpain I activity that had been inhibited by the presence of oxidant was recovered immediately by addition of the reducing agent dithiothreitol.

Calcium↗

PubMed1986

Limited autolysis of calcium-activated neutral protease (CANP): reduction of the Ca2+-requirement is due to the NH2-terminal processing of the large subunit.

Calcium-activated neutral protease (rabbit mCANP), composed of large and small subunits, was converted to a lower-Ca2+-requiring form (derived microCANP) by limited autolysis in the presence of Ca2+. The NH2-terminal regions of the two subunits of mCANP were cleaved by autolysis, but the COOH-termini remained intact after autolysis. When native mCANP or derived microCANP was dissociated into subunits, the proteolytic activity of the large subunit was reduced to 2-5% of that of the native dimeric enzyme. The Ca2+-sensitivity of one hybrid CANP reconstituted from the large subunit of derived microCANP and the small subunit of native mCANP was similar to that of derived microCANP. However, the other hybrid molecule composed of the large subunit of native mCANP and the small subunit of derived microCANP required a high concentration of Ca2+ for activity, like native mCANP. These results indicate that the Ca2+-sensitivity of derived microCANP is determined by the structural change of the large subunit resulting from loss of its NH2-terminal region. The autolysis of the small subunit apparently has no effect on the reduction of the Ca2+-requirement.

Animals↗

PubMed2000

Modification of autolysis by synthetic peptides derived from the presumptive binding domain of Staphylococcus aureus autolysin.

The autolytic cell wall hydrolase of Staphylococcus aureus, Atl, contains three highly cationic repeats in the central region of the amino acid sequence, and the repeats are presumed to have the role of binding the enzyme to some components on the cell surface. To explain the possible function of the repeats, we synthesized a number of 10- to 30-mer oligopeptides based on the Atl amino acid sequence (Thr432-Lys610) containing repeat 1, and examined their effects on the autolysis of S. aureus cells. When the peptides were added to a cell suspension of S. aureus under low ionic strength conditions, five peptides, A10, A11, A14, A16 and B9, showed immediate increases in optical density (OD) of the cell suspension accompanied by decreases in viable cell counts. After the immediate increases, the ODs for A10 and A14 changed little in the first 2 hr. In contrast, the ODs for A11 and A16 decreased rapidly. When peptide A10 was added to suspensions of heat-killed whole cells, crude cell walls and a crude peptidoglycan preparation, their ODs were increased approximately 2-fold. In contrast, the OD was not increased when the peptide was added to a suspension of pure peptidoglycan from which anionic polymers had been removed. Light microscopic and transmission electron microscopic study showed that A10 and A14 inhibited autolysis and that A11 and A16 induced autolysis earlier than the control. These results suggest strongly that the peptides adsorb to and precipitate on the anionic cell surface polymers such as teichoic acid and lipoteichoic acid via ionic interaction. The effects of peptides on the autolysis may be the results of the modification of S. aureus autolysin activities. These peptides, especially the 10-mer peptide B9 (PGTKLYTVPW) that represents the C-terminal half of A10 and N-terminal half of A11, may be important segments for Atl to bind to the cell surface.

Amino Acid Sequence↗