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International Journal of Molecular... Nov 2021Calpains are a family of Ca‑dependent cysteine proteases that participate in various cellular processes. Calpain 3 (CAPN3) is a classical calpain with unique... (Review)
Review
Calpains are a family of Ca‑dependent cysteine proteases that participate in various cellular processes. Calpain 3 (CAPN3) is a classical calpain with unique N‑terminus and insertion sequence 1 and 2 domains that confer characteristics such as rapid autolysis, Ca‑independent activation and Na activation of the protease. CAPN3 is the only muscle‑specific calpain that has important roles in the promotion of calcium release from skeletal muscle fibers, calcium uptake of sarcoplasmic reticulum, muscle formation and muscle remodeling. Studies have indicated that recessive mutations in CAPN3 cause limb‑girdle muscular dystrophy (MD) type 2A and other types of MD; eosinophilic myositis, melanoma and epilepsy are also closely related to CAPN3. In the present review, the characteristics of CAPN3, its biological functions and roles in the pathogenesis of a number of disorders are discussed.
Topics: Animals; Calpain; Enzyme Activation; Humans; Models, Biological; Muscle, Skeletal; Muscular Diseases; Organ Specificity
PubMed: 34549305
DOI: 10.3892/ijmm.2021.5036 -
BioRxiv : the Preprint Server For... Jan 2022Across the biomanufacturing industry, innovations are needed to improve efficiency and flexibility, especially in the face of challenges such as the COVID-19 pandemic....
Integrated Autolysis, DNA Hydrolysis and Precipitation Enables an Improved Bioprocess for Q-Griffithsin, a Broad-Spectrum Antiviral and Clinical-Stage anti-COVID-19 Candidate.
UNLABELLED
Across the biomanufacturing industry, innovations are needed to improve efficiency and flexibility, especially in the face of challenges such as the COVID-19 pandemic. Here we report an improved bioprocess for Q-Griffithsin, a broad-spectrum antiviral currently in clinical trials for COVID-19. Q-Griffithsin is produced at high titer in and purified to anticipated clinical grade without conventional chromatography or the need for any fixed downstream equipment. The process is thus both low-cost and highly flexible, facilitating low sales prices and agile modifications of production capacity, two key features for pandemic response. The simplicity of this process is enabled by a novel unit operation that integrates cellular autolysis, autohydrolysis of nucleic acids, and contaminant precipitation, giving essentially complete removal of host cell DNA as well as reducing host cell proteins and endotoxin by 3.6 and 2.4 log units, respectively. This unit operation can be performed rapidly and in the fermentation vessel, such that Q-GRFT is obtained with 100% yield and >99.9% purity immediately after fermentation and requires only a flow-through membrane chromatography step for further contaminant removal. Using this operation or variations of it may enable improved bioprocesses for a range of other high-value proteins in .
HIGHLIGHTS
Integrating autolysis, DNA hydrolysis and precipitation enables process simplificationAutolysis reduces endotoxin release and burden to purificationQ-Griffithsin recovered from fermentation vessel at >99.9% purity and 100% yieldQ-Griffithsin purified to anticipated clinical grade without conventional chromatographyThe resulting bioprocess is 100% disposables-compatible, scalable, and low-cost.
PubMed: 35018377
DOI: 10.1101/2021.12.30.474602 -
Journal of Dairy Science Dec 2020The importance of starter cultures to cheese manufacture and ripening is well known. Starters are inoculated into cheese milk at a level of ∼10 cfu/mL either from a... (Review)
Review
The importance of starter cultures to cheese manufacture and ripening is well known. Starters are inoculated into cheese milk at a level of ∼10 cfu/mL either from a bulk culture or using commercial direct-to-vat cultures. Before ripening, starters grow in the milk to reach populations of 10 to 10 cfu/g of curd depending on processing variables such as cook temperature, inclusion of washing steps, degree of partitioning with curds and whey, and importantly salt addition rate. Inherent strain-related properties also determine final populations in the curd following manufacture and include temperature sensitivity, salt sensitivity, presence of prophage, autolytic and permeabilization properties (which are influenced by processing steps), presence and type of cell envelope proteinase, and metabolic activity. Ripening of important industrial cheese varieties such as Cheddar, Dutch, Swiss, and Italian-type cheese varieties is characterized by extended storage under temperature-controlled conditions enabling characteristic flavor and texture development to occur. Over ripening, microbiological, biochemical and enzymatic changes occur with a decline in starter viability, release of intracellular enzymes, hydrolysis of proteins, carbohydrates and lipids, and formation of a range of volatile and nonvolatile flavor components. Recent reports suggest that starter strains may be present during the later stages of ripening and therefore their potential role needs to be reconsidered. This review will focus on our current understanding of starter viability and vitality during cheese ripening and will also review the area of starter permeabilization, autolysis, and enzyme release.
Topics: Animals; Cheese; Food Handling; Food Microbiology; Lactobacillales; Milk; Taste
PubMed: 33010919
DOI: 10.3168/jds.2020-18960 -
Biomolecular Concepts Sep 2017Amphibian metamorphosis has historically attracted a good deal of scientific attention owing to its dramatic nature and easy observability. However, the genetic... (Review)
Review
Amphibian metamorphosis has historically attracted a good deal of scientific attention owing to its dramatic nature and easy observability. However, the genetic mechanisms of amphibian metamorphosis have not been thoroughly examined using modern techniques such as gene cloning, DNA sequencing, polymerase chain reaction or genomic editing. Here, we review the current state of knowledge regarding molecular mechanisms underlying tadpole tail resorption.
Topics: Animals; Anura; Autolysis; Metamorphosis, Biological; Models, Biological; Musculoskeletal Physiological Phenomena; Phagocytosis; Reptilian Proteins; Species Specificity; Tail; Xenopus
PubMed: 28873065
DOI: 10.1515/bmc-2017-0022 -
World Journal of Microbiology &... Apr 2020Saccharomyces cerevisiae is being used for long as a rich source of proteins, sugars, nucleotides, vitamins and minerals. Autolyzed and hydrolyzed yeast biomass has...
Saccharomyces cerevisiae is being used for long as a rich source of proteins, sugars, nucleotides, vitamins and minerals. Autolyzed and hydrolyzed yeast biomass has found numerous applications in the health food industry as well as livestock feeds. Here, we have compared three lysis methods for production of yeast lysates using autolysis, plasmolysis (ethyl acetate 1.5%), and enzymatic hydrolysis (Alcalase 0.2%). The efficiency of each process was compared according to soluble solid and protein contents, cell lysis monitoring, and release of intracellular materials, cell viability and microscopic analysis. Results showed that plasmolysis by ethyl acetate was found to be more efficient compared to autolysis, with a higher recovery of yeast extract (YE) content. In comparison, the content of released solids and proteins were higher during the enzymatic hydrolysis using Alcalase compared to autolysis and plasmolysis treatments. The highest decrease in optical density of 600 nm was monitored for the hydrolyzed cells. Besides, we defined "Degree of Leakage (DL)" as a new index of the lysis process, referring to the percentage of total released proteins from the cells and it was estimated to about 65.8%, which represents an appropriate indicator of the cell lysis. The biochemical and biophysical properties of the hydrolyzed yeast product as well as its biological activity (free radical scavenging activity and bacterial binding capacity) suggest that Alcalase could be used to accelerate the lysis of yeast cells and release the valuable intracellular components used for foodstuffs, feed and fermentation media applications. Production of baker's yeast lysates using autolysis, plasmolysis, and enzymatic hydrolysis methods.
Topics: Acetates; Autolysis; Biomass; Culture Media; Fermentation; Hydrolysis; Industrial Microbiology; Saccharomyces cerevisiae; Saccharomyces cerevisiae Proteins
PubMed: 32328815
DOI: 10.1007/s11274-020-02840-3 -
BioMed Research International 2021Autolysis is an internal phenomenon following the death of an organism that leads to the degradation of tissues. In order to explore the initial stages of autolysis and...
Autolysis is an internal phenomenon following the death of an organism that leads to the degradation of tissues. In order to explore the initial stages of autolysis and attempt to establish reference standards for tissue changes after death, we studied the rapidly autolyzing tissue of the crayfish hepatopancreas. Samples from the hepatopancreas of crayfish were examined 0, 5, 10, 30, 60, and 120 minutes after death. Histological and ultrapathological examinations and evaluations and apoptotic cell counts were conducted to determine the initiation time and degree of autolysis. The results showed that autolysis in the hepatopancreas of crayfish began within 5 minutes. Initially, autolysis manifested in the swelling of hepatic tubular cells and the widening of mesenchyme. Cells undergoing autolysis showed severe organelle necrolysis. Based on these observations, tissue samples should be collected and preserved within five minutes to avoid interfering with histopathological diagnoses.
Topics: Animals; Apoptosis; Astacoidea; Autolysis; Body Size; Body Weight; Hepatopancreas; Microscopy, Electron, Transmission; Seafood; Time Factors
PubMed: 33521126
DOI: 10.1155/2021/2345878 -
Cell and Tissue Banking Sep 2021Research in neuroscience relies heavily upon postmortem human brain tissue. Cerebellar granular layer autolysis (GLA) is a surrogate marker for the quality of such...
Research in neuroscience relies heavily upon postmortem human brain tissue. Cerebellar granular layer autolysis (GLA) is a surrogate marker for the quality of such tissue and suitability for molecular analysis. GLA is associated with reduced brain tissue pH. The aim of this study was to assess correlation of GLA with premortem systemic acid-base status. This is a retrospective study in which 62 consecutive adult autopsy cases were included. Sections of cerebellum were reviewed microscopically for presence of GLA. Autolysis was graded as negative, grade 1, grade 2, and grade 3. Medical records were reviewed for arterial blood gas analysis. Postmortem interval was recorded. 23 of 62 cases showed GLA. Of the 23 patients with autolysis, 22 were acidotic and 1 was alkalotic. Of these 23 cases, 15 had metabolic acidosis, 4 had respiratory acidosis, 3 had combined acidosis and 1 had respiratory alkalosis. There was no statistically significant difference in postmortem interval between the two groups. 10 cases with grade 3 autolysis had mean pH of 7.13, 7 cases with grade 2 autolysis had mean pH of 7.23 and in 6 cases with grade 1 autolysis the mean pH was 7.2. Overall, the mean pH in patients with GLA was 7.19, and in the non-autolytic cases the mean pH was 7.28 (P < 0.05). There was no correlation between the degree of acidosis and severity of autolysis. GLA is associated with premortem systemic acidosis, and premortem systemic alkalosis is associated with the absence of GLA. Premortem acid-base status may serve as an additional quality indicator for assessment of tissue for research.
Topics: Adult; Autolysis; Autopsy; Brain; Cerebellum; Humans; Hydrogen-Ion Concentration; Retrospective Studies
PubMed: 33523332
DOI: 10.1007/s10561-021-09900-4 -
British Journal of Nursing (Mark Allen... Nov 2016Chronic wounds affect millions of people worldwide. In the UK alone, the cost of their treatment is estimated to be between £4.5bn and £5.1bn. The implementation of... (Review)
Review
Chronic wounds affect millions of people worldwide. In the UK alone, the cost of their treatment is estimated to be between £4.5bn and £5.1bn. The implementation of wound-bed preparation strategies remove the barriers to healing and wound debridement is a key component in preparing the wound bed for wound progression. This article aims to review one of the several debridement methods available to clinicians: autolytic debridement. Autolysis (i.e. autolytic debridement) uses the body's own enzymatic mechanisms to remove devitalised tissue in order to remove the barriers to healing. This review aims to provide clinicians working in wound care with a better understanding of the mechanisms and implications of autolytic debridement.
Topics: Alginates; Autolysis; Bandages, Hydrocolloid; Colloids; Debridement; Humans; Hydrogels; Wound Healing; Wounds and Injuries
PubMed: 27834531
DOI: 10.12968/bjon.2016.25.20.S40 -
International Journal of Clinical and... 2017Cancerous wound has been the difficulty of clinical treatment. Due to wound stench, symptoms like a large number of exudate and so on seriously affect the quality of...
OBJECTIVE
Cancerous wound has been the difficulty of clinical treatment. Due to wound stench, symptoms like a large number of exudate and so on seriously affect the quality of life of patients and self-esteem. Inappropriate treatment of cancerous wounds not only lead to the above-mentioned symptoms aggravating with severe infection, but also affect the treatment plan. In view of the fact that wound treatment is based on well wound bed preparation, debridement is a prerequisite for ensuring a well state of the wound bed. Therefore, exploring the most suitable way for cancer wound debridement methods, can effectively control the wound symptoms, reduce secondary infection rate, reduce complications, improve patient quality of life.
METHOD
There is a randomized controlled experiment that 22 patients with cancerous wound were nursed with autolysis debridement and mechanical debridement before and after for 2 weeks. The results of culture of secretions, the ratio of malodor, exudate and wound bed decayed tissue before and after debridement were compared.
RESULT
Through the treatment of the combination of autolysis debridement and mechanical debridement, the rate of negative conversion of secretions is increased, the control time of malodor and exudate is shortened, and the proportion of decayed tissue is reduced. The difference is statistically significant (P < 0.05).
CONCLUSION
Debridement combining autolysis debridement with mechanical debridement can effectively eliminate a large number of rotting tissue, significantly shorten the time of reducing the degree of wound infection and relieving the symptoms such as malodor and exudate, ensure the treatment of wound bed preparation, advantage to Wound healing, what's more, improve patient comfort level and quality of life.
PubMed: 31966779
DOI: No ID Found -
Experimental Biology and Medicine... Nov 2023Chitin is a biopolymer profusely present in nature and of pivotal importance as a structural component in cells. It is degraded by chitinases, enzymes naturally produced... (Review)
Review
Chitin is a biopolymer profusely present in nature and of pivotal importance as a structural component in cells. It is degraded by chitinases, enzymes naturally produced by different organisms. Chitinases are proteins enrolled in many cellular mechanisms, including the remodeling process of the fungal cell wall, the cell growth process, the autolysis of filamentous fungi, and cell separation of yeasts, among others. These enzymes also have properties with different biotechnological applications. They are used to produce polymers, for biological control, biofilm formation, and as antitumor and anti-inflammatory target molecules. Chitinases are classified into different glycoside hydrolase (GH) families and are widespread in microorganisms, including viruses. Among them, the GH18 family is highly predominant in the viral genomes, being present and active enzymes in baculoviruses and nucleocytoplasmic large DNA viruses (NCLDV), especially chloroviruses from family. These viral enzymes contain one or more GH domains and seem to be involved during the viral replication cycle. Curiously, only a few DNA viruses have these enzymes, and studying their properties could be a key feature for biological and biotechnological novelties. Here, we provide an overview of viral chitinases and their probable function in viral infection, showing evidence of at least two distinct origins for these enzymes. Finally, we discuss how these enzymes can be applied as biotechnological tools and what one can expect for the coming years on these GHs.
Topics: Humans; Chitinases; Proteins; Chitin; Biotechnology; Fungi
PubMed: 38057942
DOI: 10.1177/15353702231208408