Autolysis

High pressure homogenization (HPH) was tested for inducing autolysis in a commercial strain of Saccharomyces bayanus for winemaking. The effects on cell viability, the release of soluble proteins, glucidic colloids and amino acids in wine-like medium and the volatile composition of the autolysates were investigated after processing, in comparison with thermolysis. HPH seemed a promising technique for inducing autolysis of wine yeasts. One pass at 150MPa was the best operating conditions. Soluble colloids, proteins and free amino acids were similar after HPH and thermolysis, but the former gave a more interesting volatile composition after processing, with higher concentrations of ethyl esters (fruity odors) and lower fatty acids (potential off-flavors). This might allow different winemaking applications for HPH, such as the production of yeast derivatives for wine ageing. In the conditions tested, HPH did not allow the complete inactivation of yeast cells; the treatment shall be optimized before winemaking use.

A total of 11 of 15 Streptococcus pneumoniae culture filtrates and all five bacterial autolysates produced by cell death in the stationary phase caused slowed ciliary beating and disruption of the surface integrity of human respiratory epithelium in organ culture. This effect was inhibited by cholesterol and was heat labile and reduced by standing at room temperature but was stable at -40 degrees C. The activity was detected at the late stationary phase of culture and was associated with the presence of hemolytic activity. Gel filtration of a concentrated culture filtrate and autolysate both yielded a single fraction of approximately 50 kilodaltons which slowed ciliary beating and were the only fractions with hemolytic activity. Rabbit antiserum to pneumolysin, a sulfhydryl-activated hemolytic cytotoxin released by S. pneumoniae during autolysis, neutralized the effect of the culture filtrate on respiratory epithelium. Both native and recombinant pneumolysin caused ciliary slowing a...

Good quality fish sauce was produced from anchovies (Stolephorus spp.) that had been allowed to autolyse at pH 4 and low salt concentration. After an initial phase of rapid autolysis, the samples were neutralised, and salt was added to the normal level (250 g kg−1) before the fish sauce was separated by suction. By this method fish sauce with acceptable flavour could be produced after 2 months, whereas the normal production time is more than 6 months. This rapidly-produced fish sauce had a lower level of volatile base and acid, and a better balanced composition of essential amino acids than a first grade commercial fish sauce.

A two-step autolysis process was proposed to enhance amino acid production from spent brewer's yeast. The technique was developed based on comparative study of the dynamics of production and release of proteins and amino acids during the autolysis of a concentrated suspension (22 wt.%) and a dilute yeast cell suspension (11.25 wt.%). The results suggest that, in the concentrated yeast suspension, proteins are more effectively broken down into amino acids, but the product release rate was lower due to a lower concentration gradient across the cell membrane. Thus, a two-step process, in which a high protein conversion occurred in a concentrated cell suspension during the first 13 h period, followed by a 26 h autolysis process within a dilute cell suspension, provided a higher overall yield of amino acids compared than the single-step process. The two-step process was found to result in a 25% higher amino acid yield with a weight fraction increase from 0.4 to 0.5 g/g dry wt. Other than these findings, the effect of adding NaCl to the suspension during autolysis was also investigated. It was found that, for the autolysis conditions employed in this study, the addition of NaCl did not significantly affect the production of protein but inhibited the production of amino acids.

Gastromalacia is the acute autolytic erosion of the gastric wall. It generally occurs postmortem, and it appears as a slimy brownish black region of the wall which occurs principally in the gastric fundus. A 59-year-old woman died in the Emergency Department following a 2-day period of mild abdominal pain, vomiting, and diarrhea. A forensic autopsy was performed which revealed a rupture of the gastric fundus that had caused leakage of gastric content into the abdominal cavity. There was no macroscopic evidence of peritonitis, and the stomach wall adjacent to the rupture site showed marked thinning. The gross appearance was typical of gastromalacia. In contrast, histological observations revealed the presence of an ulcer at the site of perforation and a severe acute inflammatory reaction indicating a robust reaction with an antemortem rupture.

The behavior of the temperature-sensitive, penicillin-tolerant Escherichia coli mutant VC44 to endogenously induced autolysis by the bacteriophage phi X174 gene E product (gpE) was investigated. Expression of the cloned phi X174 lysis gene showed that cultures of strain VC44 grown at the restricted temperature were fully sensitive to endogenously induced autolysis. The results revealed that the modes of E. coli lysis induction by gpE and by penicillin differ and that the trigger mechanisms for autolysis depend greatly on the specific inducer used.

This study demonstrates post-mortem autolytic alterations in the skin at cellular and subcellular levels and identifies parameters which may assist in determining the time of death in the first few hours post-mortem. Serial skin samples from the ventral surface of the arm were taken at intervals of 3, 6, 9 and 12 h after death in 29 subjects of various ages, with no signs of skin disease; causes of death were various. Three types of tests were performed: cytochemical (hematoxylin-eosin and alcian-PAS), immunohistochemical (S-100, CEA, Cytokeratin, ASM) and ultrastructural (electron microscopy). Electron microscopy proved useful for identifying transformations which were found to be specific for each chronological step considered: reduction of intracellular glycogen in clear cells and reduction of secretory granules in dark cells are typcial signs of the first stage (3 h) after death; mitochondrial dilatation and rarefaction of cristae in clear and dark cells are typical of the second stage (6 h); rarefaction of microvilli in dark and clear cells is a sign of the last stage (12 h). Cytochemistry and immunohistochemistry supply useful information — not for all the chronological stage considered here, but for individual phases (3 h for hematoxylin-eosin and 6 h for alcian-PAS). However, it is particularly important to use the results from all such techniques simultaneously, so that the question of the exact time of death within the first 12 h post-mortem may be more accurately answered. Diese Untersuchung zeigt postmortale autolytische Veränderungen in der Haut auf zellulärer und subzellulärer Ebene und identifiziert Parameter, welche helfen können, die Zeit des Todes in den ersten Stunden postmortem zu bestimmen. Hautproben von der Beugeseite des Arms wurden, 3, 6, 9 und 12. Stunden nach dem Tode von insgesamt 29 Leichen entnommen (verschiedene Altersklassen, keine Zeichen für Hauterkrankungen, verschiedene Todesursachen). Drei Arten der Untersuchungen wurden durchgeführt: zytochemisch (Hematoxylin-Eosin and Alcian-PAS), immunhistochemisch (S-100, CEA, Cytokeratin, ASM) und ultrastrukturell (Elektronenmikroskopie). Die Elektronenmikroskopie erwies sich als nützlich für die Identifizierung von Transformationen die für jeden chronologischen Schritt spezifisch waren: Reduktion des intrazellulären Glykogens in hellen Zellen und Reduktion der sekretorischen Granula in dunklen Zellen sind typische Zeichen für die erste Phase (3 Stunden) nach dem Tode; mitochondriale Dilatation und Rarifizierung der Cristae in hellen und dunklen Zellen sind typisch für die 2. Phase (6 Stunden); Rarifizierung der Microvilli in dunklen und hellen Zellen sind typisch für die 3. Phase (9 Stunden) und Kernpyknose von dunklen und hellen Zellen ist ein Zeichen der letzten Phase (12 Stunden). Zytochemie und Immunhistochemie sorgen für eine nützliche Information — dies gilt nicht für alle chronologischen Stadien, welche hier einbezogen wurden, aber für individuelle Phasen (3 Stunden für Hematoxylin-Eosin und 6 Stunden für Alcian-PAS). Es ist jedoch besonders wichtig, die Resultate von allen solchen Techniken simultan einzubeziehen, so daß die Frage der exakten Todeszeit innerhalb der ersten 12 Stunden postmortem genauer beantwortet werden kann.

Aims:  Identification and biotyping of lactic acid bacteria (LAB) isolated from raw-milk Pecorino cheese manufactured in the Marche region (central Italy) for selection of suitable starter cultures or adjuncts.Methods and Results:  Preliminary characterization with morphological and biochemical assays were undertaken for 112 Gram-positive and catalase-negative isolates. Unequivocal identification of the isolates was obtained through restriction analysis of the amplified 16S rRNA gene and sequencing of 360–380 bp amplicons. Fifty-nine isolates belonging to LAB species generally recognized as safe and potentially utilized as starters or flavour-producing adjuncts were preselected and tested for their acidifying, proteolitic and autolytic activities. Fifty-five of these isolates were also subject to RAPD (randomly amplified polymorphic DNA) fingerprinting and unweighted pair-group method with arithmetic averages (UPGMA) cluster analysis for the estimation of genotypic intra-species variation. As a result, in Pecorino cheese, a heterogeneous lactic acid bacteria population, which includes strains with metabolic characteristics of technological interest, was characterized.Conclusions:  The polyphasic approach proposed allows the bacterial ecology of Pecorino cheese to be investigated and allows to assess the potential role of autochthonous LAB strains for the dairy industry.Significance and Impact of the Study:  The great economic importance of Pecorino cheese encouraged a deeper knowledge of its microbiota, which is known to influence the peculiar sensory properties of this cheese, also in view of its exploitation.

Compositions and yield of lipids extracted from hepatopancreas of Pacific white shrimp (Litopenaeus vannamei) subjected to autolysis at 60 C for different times (0, 30, 60, 90 120 and 150 min) were investigated. Extraction yield increased from 7.4% to 8.8% as autolysis time increased from 0 to 150 min. Coincidental increase in total carotenoid content was obtained with increasing autolysis time (p < 0.05). The increases in thiobarbituric acid-reactive substances (TBARS) and p-anisidine value (AV) of lipids were noticeable when autolysis time increased (p < 0.05). However, no changes in free fatty acid (FFA) content were observed within the first 60 min of autolysis (p > 0.05), but subsequently increased up to 150 min. (p < 0.05). No differences in fatty acid profiles of lipids extracted from hepatopancreas without and with 60 min prior autolysis were observed. Lipids extracted contained docosahexaenoic acid (DHA; C22:6(n  3)) as the most abundant fatty acid, followed by eicosapentaenoic acid (EPA; C20:5(n  3)). Therefore, prior autolysis at 60 C for 60 min increased the extraction yield without negative effect on lipid quality.

The goals of this study were to determine 1) the expression of calpain isoforms in rabbit renal proximal tubules (RPT); 2) calpain autolysis and translocation, and calpastatin levels during RPT injury; and 3) the effect of a calpain inhibitor (PD-150606) on calpain levels, mitochondrial function, and ion transport during RPT injury. RT-PCR, immunoblot analysis, and FITC-casein zymography demonstrated the presence of only μ- and m-calpains in rabbit RPT. The mitochondrial inhibitor antimycin A decreased RPT μ- and m-calpain and calpastatin levels in conjunction with cell death and increased plasma membrane permeability. No increases in either μ- or m-calpain were observed in the membrane nor were increases observed in autolytic forms of either μ- or m-calpain in antimycin A-exposed RPT. PD-150606 blocked antimycin A-induced cell death, preserved calpain levels in antimycin A-exposed RPT, and promoted the recovery of mitochondrial function and active Na+ transport in RPT after hypoxia...

Two deteriorative pathways (autolysis and microbiological activity) were studied in farmed Coho salmon (Oncorhynchus kisutch) for 24 days during chilled storage. These changes were assessed by nucleotide degradation (determination of adenosine 5′-triphosphate and its degradation compounds) and biochemical (pH; content of total volatile base-nitrogen; trimethylamine-nitrogen, TMA-N; histamine) and microbial (total aerobe mesophiles, TAM; coliforms) indices related to bacterial activity. An important nucleotide degradation could be assessed according to the fast inosine 5′-monophosphate formation, followed by degradation into inosine and hypoxanthine; the K value was found to be an accurate tool for the measurement of quality loss throughout the whole experiment. Regarding bacterial activity, contents of TMA-N and histamine and TAM counts assessment showed sharp increases after the end of the microbial lag phase (12–17 days); however, values obtained for histamine content and TAM growth remained below acceptable security limits throughout the whole experiment.

ZusammenfassungDer Beitrag zeigt, wie die von Niklas Luhmann entwickelte Systemtheorie für eine empirische Analyse von Lärm und lärmerzeugenden Mechanismen geöffnet werden kann. Die dem Artikel zugrunde liegende analytische Strategie ist die Beobachtung der Operationen, die ein soziales System konstituieren. Eine entsprechende Analyse macht auf das Fehlen von Anschlusskommunikation und auf die aktive Produktion von Lärm (verstanden als Operationen ohne Anschlüsse) aufmerksam, d. h. sie öffnet die Analyse für systemische Autolysis (Selbstauflösung). Darauf aufbauend wird eine operationale Analyse organisationaler Kommunikation durchgeführt, die die Leitdifferenz Ereignis/Rekursivität benutzt. Der Artikel ist in sechs Abschnitte gegliedert. Nach der Einleitung wird eine operational-analytische Strategie skizziert (I). Danach wird das Konzept der Autolysis vorgestellt (II). Eine Fallstudie über das Entscheidungsverhalten in einer Organisation des Gesundheitswesens liefert dann Beispiel...