Impacts on the immune system of Cyprinus carpio exposure with a mixed algal extract against Aeromonas hydrophila

This study evaluates the influence of mixed algal extract (<i>Chlorella vulgaris</i>, <i>Euglena viridis</i> and <i>Spirulina platensis</i>) on common carp <i>Cyprinus Carpio</i>, which infected infect with bacterial pathogen <i>Aeromonas hydrophila</i>. <i>C. carpio</i> was administered intraperitoneally with various doses such as methanol extract (0, 0,1, 1, 10 and 100 mg/kg). The immunological parameters of fish blood and serum samples (Neutrophil activity, Lysozyme activity, Serum myeloperoxidase intensity, Serum bactericidal activity, and Serum antiprotease activity) were investigated at 7, 14, 21, and 28 days of post-immunization. Fish had been tested by virulent<i> A. hydrophila</i> for 30 days after treatment and 14 days after infection were identified with mortalities. The findings showed that neutrophil levels, lysozyme activity, serum bactericidal activity, myeloperoxidase activity, and serum antiprotease activity significantly enhanced (p<0.05) compared to untreated control. Mixed dietary algae at 1 and 10 mg/kg levels demonstrated slightly (p<0.05) higher relative percentage survival (90 percent) than control against <i>A. hydrophila</i> disease infection. Results indicated that mixed algal extract in <i>C. carpio</i> positively impacts non-specific immune parameters and boosts disease tolerance to <i>A. hydrophila</i> infections.

PHYTOCHEMICAL SCREENING, IN VITRO ANTIOXIDANT AND ANTI PROTEASE ACTIVITIES OF METHANOLIC EXTRACT OF NILGIRIANTHUS HEYNEANUS NEES STEM

Objective: The aim of this study was focussed on phytochemical screening, in vitro antioxidant and antiprotease activities of methanolic extract of Nilgirianthus heyneanus stem. Methods: The stem of the plant was washed thoroughly, shade dried and coarsely powdered. The powdered material of Niligirianthus heyneanus stem was extracted with methanol using soxhlet apparatus. Preliminary phytochemical screening for carbohydrates, proteins, alkaloids, phytosteroids, flavonoids, glycosides, polyphenolics, saponins, tannins was done by following standard procedure. In vitro antioxidant activities of methanolic extract were assessed using DPPH, ABTS and total antioxidant capacity. In vitro anti-protease activity of the plant was analysed using trypsin as an enzyme and BAEE (N-benzoyl-L-arginine ethyl ester) as a substrate. Results: The results showed that phytochemicals such as carbohydrates, proteins and amino acids, flavonoids, glycosides, tannins and polyphenolics are present in the methanolic extract of Niligirianthus heyneanus stem. The in vitro antioxidant and antiprotease activities of Niligirianthus heyneanus stem clearly showed that the plant has antioxidant and antiprotease activity. Conclusion: From this work, it can be concluded that Niligirianthus heyneanus stem has the potential to be a strong antioxidant and protease inhibitor.

α1-Antitrypsin: Key Player or Bystander in Acute Respiratory Distress Syndrome?

Acute respiratory distress syndrome is characterized by hypoxemia, altered alveolar–capillary permeability, and neutrophil-dominated inflammatory pulmonary edema. Despite decades of research, an effective drug therapy for acute respiratory distress syndrome remains elusive. The ideal pharmacotherapy for acute respiratory distress syndrome should demonstrate antiprotease activity and target injurious inflammatory pathways while maintaining host defense against infection. Furthermore, a drug with a reputable safety profile, low possibility of off-target effects, and well-known pharmacokinetics would be desirable. The endogenous 52-kd serine protease α1-antitrypsin has the potential to be a novel treatment option for acute respiratory distress syndrome. The main function of α1-antitrypsin is as an antiprotease, targeting neutrophil elastase in particular. However, studies have also highlighted the role of α1-antitrypsin in the modulation of inflammation and bacterial clearance. In light of the current SARS-CoV-2 pandemic, the identification of a treatment for acute respiratory distress syndrome is even more pertinent, and α1-antitrypsin has been implicated in the inflammatory response to SARS-CoV-2 infection.

Post-Translational Modifications of Circulating Alpha-1-Antitrypsin Protein

Alpha-1-antitrypsin (AAT), an acute-phase protein encoded by the SERPINA1 gene, is a member of the serine protease inhibitor (SERPIN) superfamily. Its primary function is to protect tissues from enzymes released during inflammation, such as neutrophil elastase and proteinase 3. In addition to its antiprotease activity, AAT interacts with numerous other substances and has various functions, mainly arising from the conformational flexibility of normal variants of AAT. Therefore, AAT has diverse biological functions and plays a role in various pathophysiological processes. This review discusses major molecular forms of AAT, including complex, cleaved, glycosylated, oxidized, and S-nitrosylated forms, in terms of their origin and function.

EVALUATION OF BONE RESORPTIVE POTENTIAL IN THE TREATMENT OF GENERALIZED PERIODONTITIS

The aim: Is to study dynamics of resorptive potential of bone tissue by indicators of protease and antiprotease systems in the process of treatment generalized periodontitis in patients with age-related osteoporosis and without osteoporotic changes in the skeleton. Materials and methods: In 102 patients, before the start of treatment of generalized periodontitis and in 2, 4 and 12 weeks, the concentration of pro-resorbing matrix metalloproteinases: stromelysin (MMP-3), collagenase (MMP-8) and gelatinase (MMP-9) were determined in blood plasma and mixed oral fluid. The antiresorbtive potential of bone tissue was evaluated by the concentration of a tissue inhibitor of metalloproteinases (TIMP-1) in plasma. The general antiprotease activity was determined by the activity of α1-antitrypsin (α1-AT) and α2-macroglobulin (α2-MG). Results: The most significant changes are recorded for the content of MMP-9 in blood and oral fluid, regardless of the presence of systemic disorders of bone metabolism (P < 0.05). Concentration of MMP-8 is significantly increased in blood plasma and oral fluid in accordance with the severity of the disease and in the course of treatment (P <0.05). The observed increase in the ratio of MMP-8 to TIMP-1 and MMP-9 to TIMP-1 in patients with generalized periodontitis, complicated by systemic osteoporosis (P < 0.05), indicates an imbalance of the protease-antiprotease system. Conclusions: The resorptive potential of bone tissue in patients with generalized periodontitis allows us to correctly choose treatment tactics and to prevent the development of complications.

Pathophysiology of Emphysema

Pulmonary emphysema is part of pathological condition in chronic obstructive pulmonary disease (COPD) which is characterized by lung parenchymal destruction. Morphology classification of emphysema had been made according to histologic structure in pathology. There were some causes known to be the culprit of emphysema; one that caught most attention is protease-antiprotease activity from cigarette smoke exposure. Destructive effect of emphysema gives disturbance of lung function in expiration (obstruction). The primary mechanism is elastic recoil reduction which causes air trapping, lung volume increase, lung compliance increase and airways that is susceptible to collapse. Hyperinflation in emphysema causes some disadvantages which complicate inspiration and give a dyspnea sensation. Equal pressure point drop in emphysema happens because of elastic recoil reduction. This drop may cause early airway closure. (J Respir Indo 2019; 39(1): 60-9)

An antibody that prevents serpin polymerisation acts by inducing a novel allosteric behaviour

Serpins are important regulators of proteolytic pathways with an antiprotease activity that involves a conformational transition from a metastable to a hyperstable state. Certain mutations permit the transition to occur in the absence of a protease; when associated with an intermolecular interaction, this yields linear polymers of hyperstable serpin molecules, which accumulate at the site of synthesis. This is the basis of many pathologies termed the serpinopathies. We have previously identified a monoclonal antibody (mAb4B12) that, in single-chain form, blocks α1-antitrypsin (α1-AT) polymerisation in cells. Here, we describe the structural basis for this activity. The mAb4B12 epitope was found to encompass residues Glu32, Glu39 and His43 on helix A and Leu306 on helix I. This is not a region typically associated with the serpin mechanism of conformational change, and correspondingly the epitope was present in all tested structural forms of the protein. Antibody binding rendered β-sheet A — on the opposite face of the molecule — more liable to adopt an ‘open’ state, mediated by changes distal to the breach region and proximal to helix F. The allosteric propagation of induced changes through the molecule was evidenced by an increased rate of peptide incorporation and destabilisation of a preformed serpin–enzyme complex following mAb4B12 binding. These data suggest that prematurely shifting the β-sheet A equilibrium towards the ‘open’ state out of sequence with other changes suppresses polymer formation. This work identifies a region potentially exploitable for a rational design of ligands that is able to dynamically influence α1-AT polymerisation.

Expression of SERPINA3s in cattle: focus on bovSERPINA3-7 reveals specific involvement in skeletal muscle

α 1 -Antichymotrypsin is encoded by the unique SERPINA3 gene in humans, while it is encoded by a cluster of eight closely related genes in cattle. BovSERPINA3 proteins present a high degree of similarity and significant divergences in the reactive centre loop (RCL) domains which are responsible for the antiprotease activity. In this study, we analysed their expression patterns in a range of cattle tissues. Even if their expression is ubiquitous, we showed that the expression levels of each serpin vary in different tissues of 15-month-old Charolais bulls. Our results led us to focus on bovSERPINA3-7, one of the two most divergent members of the bovSERPINA3 family. Expression analyses showed that bovSERPINA3-7 protein presents different tissue-specific patterns with diverse degrees of N -glycosylation. Using a specific antibody raised against bovSERPINA3-7, Western blot analysis revealed a specific 96 kDa band in skeletal muscle. BovSERPINA3-7 immunoprecipitation and mass spectrometry revealed that this 96 kDa band corresponds to a complex of bovSERPINA3-7 and creatine kinase M-type. Finally, we reported that the bovSERPINA3-7 protein is present in slow-twitch skeletal myofibres. Precisely, bovSERPINA3-7 specifically colocalized with myomesin at the M-band region of sarcomeres where it could interact with other components such as creatine kinase M-type. This study opens new prospects on the bovSERPINA3-7 function in skeletal muscle and promotes opportunities for further understanding of the physiological role(s) of serpins.