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The self‐organization of microbes into biofilms provides multiple benefits including tolerance to mechanical stress and resistance to immune defences and antibiotics. Coupled to a compromised mucociliary function, these traits have dire consequences in cystic fibrosis patients – persistent infections are the main reason for morbidity and mortality. Thereby, disease progression is associated with universal colonization by Pseudomonas aeruginosa, which selects for a slimy phenotype to adapt to the lung microenvironment. Recognizing this, drug‐delivery vehicles that break down the mucoid extracellular matrix made of alginate are designed to enable better penetration and biofilm dispersion. In parallel, a protective layer responds to the proteolytic activity of the pathogen and thus controls drug availability. To realize this architecture, silica nanoparticles are loaded with imipenem, and then coated with elastin and alginate lyase in a layer‐by‐layer fashion using ultrasound. The nanoscale formulations eradicate up to 80% of the total biomass and reduce the bacterial viability in biofilms by 3 logs, considerably outperforming the bulk antibiotic in vitro, whereby the effects are correlated to changes in the viscoelasticity. Furthermore, the stimuli‐responsive nanocarriers are safe and effective in animal models of P. aeruginosa infection, presenting a considerable therapeutic promise in the challenging context of lung diseases. [ABSTRACT FROM AUTHOR]

• This is the first study on phytochemical profiling and enzyme inhibitory activities of C. corymbosum extracts. • Chlorogenic acid, rutin, and quercitrin were major compounds in the extracts. • The extracts have antioxidant and cytotoxic activities. • The extracts showed significant inhibitory effects on α-glucosidase and thioredoxin reductase. Cardopatium corymbosum (L.) Pers. (Asteraceae) has been used ethnomedicinally against intestinal worms, for skin diseases, and in wound treatment. This study investigated the phytochemical profile and biological activities of ethanol (CCE), methanol (CCM), and water (CCW) extracts of C. corymbosum. LC-MS/MS analysis revealed the presence of various metabolites. Antioxidant activities of the extracts were determined by measuring 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activities and ferric-reducing power (FRAP). The cytotoxicity against human breast cancer (MCF-7) and human umbilical vein endothelial (HUVEC) cell lines were determined using the (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) (MTT) assay. Enzyme inhibitory activities against α-amylase, α-glucosidase, acetylcholinesterase (AChE), butyrylcholinesterase (BChE), thioredoxin reductase (TrxR), and elastase were measured to assess potential antidiabetic, anti-Alzheimer's, anticancer, and antiaging/wound healing effects. CCM exhibited the highest total phenolic and flavonoid contents and antioxidant activity. Fumaric acid, chlorogenic acid, vanillic acid, and rutin were identified as major components in CCM. C. corymbosum extracts, particularly those from CCE and CCM, displayed significant cytotoxicity on MCF-7 cells while exhibiting lower toxicity towards HUVEC cells. Notably, CCM demonstrated potent α-glucosidase inhibitory activity, while no α-amylase inhibition was observed. All extracts showed weak inhibition of AChE, BChE, and elastase. Conversely, the extracts displayed moderate TrxR inhibitory activity. These findings suggest that C. corymbosum possesses significant antioxidant, enzyme inhibitory, and cytotoxic potential, highlighting its promise as a source of novel bioactive molecules. [ABSTRACT FROM AUTHOR]

In the aging process, skin morphology might be affected by wrinkle formation due to the loss of elasticity and resilience of connective tissues linked to the cleavage of elastin by the enzymatic activity of elastase. Little information is available about the structural requirements to efficiently inhibit elastase 1 (EC 3.4.21.36) expressed in skin keratinocytes. In this study, a structure-based approach led to the identification to the pharmacophoric hypotheses that described the main structural requirements for binding to porcine pancreatic elastase as a valuable tool for the development of skin therapeutic agents due to its similarity with human elastase 1. The obtained models were subsequently refined through the application of computational alanine-scanning mutagenesis to evaluate the effect of single residues on the binding affinity and protein stability; in turn, molecular dynamic simulations were carried out; these procedures led to a simplified model bearing few essential features, enabling a reliable collection of chemical features for their interactions with elastase. Then, a virtual screening campaign on the in-house library of synthetic compounds led to the identification of a nonpeptide-based inhibitor (IC50 = 60.4 µM) belonging to the class of N-substituted-1H-benzimidazol-2-yl]thio]acetamides, which might be further exploited to obtain more efficient ligands of elastase for therapeutic applications. [ABSTRACT FROM AUTHOR]

Peptides conjugated with organic molecules can be useful tools for the development of novel bioactive compounds. The lysine side‐chain is an interesting functional group to act as a linker to connect small molecules in peptide conjugates. Herein we present the design and synthesis of four Safirinium derivatives, their facile conjugation to lysine in solid phase, and incorporation in peptides. The compounds differing in the functionalization of nitrogen N2 of the condensed triazolyl moiety are novel building blocks to design conjugates to perform structure‐activity relationship studies of elastase inhibitors. Consequently, structural investigations of the lysine building blocks and of the corresponding peptide conjugates were performed. We present herein the potential of the Safirinium analogs to act as elastase inhibitors in assays performed on porcine pancreas elastase. [ABSTRACT FROM AUTHOR]

Chios mastic gum (CMG), the resin of Pistacia lentiscus var. Chia, is a product with great ethnopharmacological and economic significance. This study attempts to investigate, for the first time, the activity of CMG, its fractions and isolated compounds against specific enzymes, which play pivotal roles in the degradation of proteins contained in skin connective tissue. Initially, crude CMG was subjected to extraction, fractionation and isolation through different chromatographic techniques to obtain the acidic and neutral fraction of terpenes. Additionally, the characteristic and major active triterpene acids of CMG, masticadienonic and isomasticadienonic acids (MNA, IMNA) were isolated in pure form. All samples were analysed by means of High-Performance Thin-Layer Chromatography (HPTLC) with four distinct development systems to obtain their constituents' profile. Finally, samples were tested for their ability to inhibit the elastase and collagenase enzymes. According to our findings, for collagenase, a mixture of MNA and IMNA demonstrated the most potent activity with an IC50 value of 31.07 μg/mL, while for elastase CMG's acidic fraction provided the most promising results with an IC50 value of 17.30 μg/mL. Overall, these results attempt to fill the gap in scientific knowledge about the use of CMG and its constituents in skincare and cosmetic products. [ABSTRACT FROM AUTHOR]

Background: Serine proteases represent over 1% of all proteins in humans. This family of proteins is found on cell surfaces, subcellular organelles like lysosomes or mitochondria, within the nucleus and the protoplasm. Among them, trypsin, chymotrypsin and elastase have aroused great interest because of their numerous functions in pathophysiological processes. Altered expression of these enzymes in experimental animal models and humans has been related to various pathologies, like developmental defects, metabolic dysfunctions, cancer, peripheral vascular diseases and infectious diseases. Trypsin and chymotrypsin-like proteases activate, or less oftentimes inactivate, numerous substrates, together with growth factors, receptors, adhesion molecules, angiogenic factors and metalloproteases. Among these substrates, a number of them are key factors in cancer progression, metastasis and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease. Elastin-degrading enzyme- elastase, slowly damages elastin over the lifetime of an organism. The physiological processes triggered by elastase leads to the progression of different conditions such as cancer, metabolic syndrome, pulmonary emphysema, atherosclerosis, and chronic obstructive pulmonary disease. These serine proteases are currently considered to be targets for the development of new potent therapeutics. Short conclusion: The cumulative knowledge that outlined the physiological functions and pathological implications of these proteases and the proposed strategies to regulate a number of their activities and their targeting for therapeutic application and validation in selected disease states are highlighted. These should enhance our appreciation of their roles in aetiology of some diseases as well as the chemotherapeutic benefits of their inhibition or modulation. [ABSTRACT FROM AUTHOR]

We have analysed the in silico potential of bioactive peptides from cheese whey, the most relevant by-product from the dairy industry, to bind into the active site of collagenase and elastase. The peptides generated from the hydrolysis of bovine β-lactoglobulin with three proteases (trypsin, chymotrypsin, and subtilisin) were docked onto collagenase and elastase by molecular docking. The interaction models were ranked according to their free binding energy using molecular dynamics simulations, which showed that most complexes presented favourable interactions. Interactions with elastase had significantly lower binding energies than those with collagenase. Regarding the interaction site, it was found that four bioactive peptides were positioned in collagenase's active site, while six were found in elastase's active site. Among these, the most we have found one promising collagen-binding peptide produced by chymotrypsin and two for elastase, produced by subtilisin and chymotrypsin. These in silico results can be used as a tool for designing further experiments aiming at testing the in vitro potential of the peptides found in this work. [ABSTRACT FROM AUTHOR]

Objective: Abdominal aortic aneurysms (AAA) are a common critical cardiovascular disease with high morbidity and mortality rates. Quercetin, a flavonoid and a traditional Chinese medicinal ingredient commonly found in vegetables and fruits, has a favorable inhibitory effect on experimental aneurysms without side effects. This study aimed to model elastase AAA and investigate the mechanism of action of quercetin in alleviating AAA. Methods: Quercetin was administered at a dose of 60 mg/kg once daily starting on the day of AAA induction for 5 weeks. The therapeutic effect of quercetin on AAA was demonstrated using biochemical assays, observation of pathological tissue sections, Elastic Van Gieson staining, immunohistochemistry, molecular docking simulations, and Western blot protein validation. Results: Our study showed that quercetin treatment significantly alleviated abdominal aortic vessel wall dilatation, elastin degradation and adhesion to surrounding tissues caused by elastase. Additionally, quercetin significantly inhibited the mRNA and protein expression of the receptor for advanced glycation end products(RAGE)/ phosphatidylinositol 3-kinase (PI3K)/ phosphatidylinositol 3-kinase (AKT) / mammlian target of rapamycin (mTOR) pathway. Conclusion: Quercetin can alleviate abdominal aortic injury caused by elastase via RAGE/PI3K/AKT/mTOR and provide experience in clinical use. [ABSTRACT FROM AUTHOR]

Introduction: Traditional Eastern medicine (TEM) is becoming increasingly more popular in highly developed Western countries as an alternative form of supporting health and body care. Many herbs used in this medical practice possess antioxidant, anti-inflammatory and immunomodulatory effects. The skin aging process may progress with age, when collagen and elastin fibers gradually decrease. Excessive exposure to UV radiation, resulting in an increase in the production of free radicals, leads to damage at the molecular level to numerous structures in the body including the acceleration of skin aging. Material and methods: The content of polyphenolic compounds (among others: phenolic acids and flavonoids), antioxidant potential (ABTS, DPPH and FRAP assays) as well as the influence on the activity of enzymes, collagenase and elastase, were determined in infusions obtained from Gynostemma pentaphyllum, Tinospora cordifolia, Astragalus membranaceus, Codonopsis pilosula, Asparagus racemosus and Ocimum sanctum. Results: The highest content of polyphenolic compounds and the strongest antioxidant properties were observed in the infusions obtained from the O. sanctum herb, while the greatest ability to inhibit collagenase and elastase was observed in the infusions obtained from the T. cordifolia leaves. Conclusions: Infusions from the O. sanctum herb and T. cordifolia leaves may have a potentially beneficial effect on the skin and may be used in anti-aging formulations.

Abstract Background Serine proteases represent over 1% of all proteins in humans. This family of proteins is found on cell surfaces, subcellular organelles like lysosomes or mitochondria, within the nucleus and the protoplasm. Main body of the abstract Among them, trypsin, chymotrypsin and elastase have aroused great interest because of their numerous functions in pathophysiological processes. Altered expression of these enzymes in experimental animal models and humans has been related to various pathologies, like developmental defects, metabolic dysfunctions, cancer, peripheral vascular diseases and infectious diseases. Trypsin and chymotrypsin-like proteases activate, or less oftentimes inactivate, numerous substrates, together with growth factors, receptors, adhesion molecules, angiogenic factors and metalloproteases. Among these substrates, a number of them are key factors in cancer progression, metastasis and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) disease. Elastin-degrading enzyme- elastase, slowly damages elastin over the lifetime of an organism. The physiological processes triggered by elastase leads to the progression of different conditions such as cancer, metabolic syndrome, pulmonary emphysema, atherosclerosis, and chronic obstructive pulmonary disease. These serine proteases are currently considered to be targets for the development of new potent therapeutics. Short conclusion The cumulative knowledge that outlined the physiological functions and pathological implications of these proteases and the proposed strategies to regulate a number of their activities and their targeting for therapeutic application and validation in selected disease states are highlighted. These should enhance our appreciation of their roles in aetiology of some diseases as well as the chemotherapeutic benefits of their inhibition or modulation.

Abstract Background PD-1/PD-L1 blockade has become a powerful method to treat malignant tumors. However, a large proportion of patients still do not benefit from this treatment, due to low tumor immunogenicity and low tumor penetration of the agents. Recently, neutrophil elastase has been shown to induce robust tumor immunogenicity, while the insufficient enzyme activity at the tumor site restricted its anti-tumor application. Here, we designed polyethyleneimine-modified neutrophil elastase (PEI-elastase) loaded with PD-L1small interfering RNA (PD-L1 siRNA) for improving enzymatic activity and delivering siRNA to tumor, which was expected to solve the above-mentioned problems. Results We first demonstrated that PEI-elastase possessed high enzymatic activity, which was also identified as an excellent gene-delivery material. Then, we synthesized anti-tumor lipopolymer (P-E/S Lip) by encapsulating PEI-elastase and PD-L1siRNA with pH-responsive anionic liposomes. The P-E/S Lip could be rapidly cleaved in tumor acidic environment, leading to exposure of the PEI-elastase/PD-L1 siRNA. Consequently, PEI-elastase induced powerful tumor immunogenicity upon direct tumor killing with minimal toxicity to normal cells. In parallel, PEI-elastase delivered PD-L1siRNA into the tumor and reduced PD-L1 expression. Orthotopic tumor administration of P-E/S Lip not only attenuated primary tumor growth, but also produced systemic anti-tumor immune response to inhibit growth of distant tumors and metastasis. Moreover, intravenous administration of P-E/S Lip into mice bearing subcutaneous tumors leaded to an effective inhibition of established B16-F10 tumor and 4T1 tumor, with histological analyses indicating an absence of detectable toxicity. Conclusions In our study, a protease-based nanoplatform was used to cooperatively provoke robust tumor immunogenicity and down-regulate PD-L1 expression, which exhibited great potential as a combination therapy for precisely treating solid tumors. Graphical Abstract

The tamarillo, or Solanum betaceum, recognized for its comprehensive nutritional profile, has long been valued for its diverse ethnobotanical uses. This study delves into the potential therapeutic applications of S. betaceum by analyzing its polyphenolic content (TPC), total flavonoid content (TFC), anti-skin aging activities against key enzymes like elastase, tyrosinase, and hyaluronidase, and its cytotoxic effects on oral carcinoma cells. Extracts from the seeds, pulp, and peel of red and yellow fruits were prepared using methanol, ethanol, and acetone. The highest TPC was found in the methanol extract from red fruit seeds (9.89 mg GAE/g), and the highest TFC was found in the methanol extract of yellow fruit peel (3.02 mg QUE/g). Some of these extracts significantly inhibited skin aging-associated enzymes with the red fruit seed extract (100 μg/mL) showing up to 50.4% inhibition of tyrosinase. Additionally, the red fruit seed extract obtained using methanol demonstrated potential anticancer effects against Ca9-22 oral carcinoma cells by inhibiting cell survival, migration, and proliferation as well as inducing apoptosis. These results underscore the potential of S. betaceum fruit extracts, especially from red fruit seeds, as promising agents for anti-skin aging and anticancer applications, meriting further exploration for therapeutic uses. [ABSTRACT FROM AUTHOR]

Introduction: Schinus terebinthifolia (ST) is a plant belonging to the cashew family Anacardiaceae, native to subtropical and tropical South America. ST is commonly called Brazil pepper and aroeira. Several reports have been made on the biological activities of ST, but studies on leaf extracts, especially lectins, have mainly been reported. Objectives: Our study analyzed the active compounds, antioxidant activities, xanthine oxidase inhibitory, elastase inhibitory, and tyrosinase inhibitory activities of S. terebinthifolia (ST) bark extract. Results: Hot water extracts showed the strongest electron donating ability (84.46%) and tyrosinase inhibitory activity (67.1%). Eighty percent ethanol extract showed the highest reducing power, total phenolic, xanthine oxidase (91.7%) and elastase inhibitory ability (85.44%). Catechin, α–amyrin, β–amyrone and 11-Oxo-.α-amyrin were identified through HPLC and GCMS analysis, while eighty percent extract contained the highest amount of catechin. Catechin, α–amyrin, and β–amyrone are considered to be the main xanthine oxidase inhibitors, while β–amyrone is considered to be the main inhibitor of xanthine oxidase and elastase. Conclusion: Through this study, we reported the basic information that S. terebinthifolia bark extract was used in folk medicine as an anti-inflammatory, anti-gout, and skin disease improvement material. S. terebinthifolia bark extract could be used as an anti-gout natural drug or cosmetic material. [ABSTRACT FROM AUTHOR]

Background: PD-1/PD-L1 blockade has become a powerful method to treat malignant tumors. However, a large proportion of patients still do not benefit from this treatment, due to low tumor immunogenicity and low tumor penetration of the agents. Recently, neutrophil elastase has been shown to induce robust tumor immunogenicity, while the insufficient enzyme activity at the tumor site restricted its anti-tumor application. Here, we designed polyethyleneimine-modified neutrophil elastase (PEI-elastase) loaded with PD-L1small interfering RNA (PD-L1 siRNA) for improving enzymatic activity and delivering siRNA to tumor, which was expected to solve the above-mentioned problems. Results: We first demonstrated that PEI-elastase possessed high enzymatic activity, which was also identified as an excellent gene-delivery material. Then, we synthesized anti-tumor lipopolymer (P-E/S Lip) by encapsulating PEI-elastase and PD-L1siRNA with pH-responsive anionic liposomes. The P-E/S Lip could be rapidly cleaved in tumor acidic environment, leading to exposure of the PEI-elastase/PD-L1 siRNA. Consequently, PEI-elastase induced powerful tumor immunogenicity upon direct tumor killing with minimal toxicity to normal cells. In parallel, PEI-elastase delivered PD-L1siRNA into the tumor and reduced PD-L1 expression. Orthotopic tumor administration of P-E/S Lip not only attenuated primary tumor growth, but also produced systemic anti-tumor immune response to inhibit growth of distant tumors and metastasis. Moreover, intravenous administration of P-E/S Lip into mice bearing subcutaneous tumors leaded to an effective inhibition of established B16-F10 tumor and 4T1 tumor, with histological analyses indicating an absence of detectable toxicity. Conclusions: In our study, a protease-based nanoplatform was used to cooperatively provoke robust tumor immunogenicity and down-regulate PD-L1 expression, which exhibited great potential as a combination therapy for precisely treating solid tumors. [ABSTRACT FROM AUTHOR]

Aneurysms are clinical entities that can develop and affect human aorta; and although in most cases they have an asymptomatic course, these pathological dilatations can lead to a lethal outcome when rupture occurs, thus the establishment of predictors is crucial for death prevention. Essential events that take place in the vessel wall have been identified and described, such as inflammation, proteolysis, smooth muscle cell apoptosis, angiogenesis, and vascular remodeling. Porcine and ovine models have been useful for the development and evaluation of endovascular devices of the aorta. However, since the worldwide introduction and adoption of these minimally invasive techniques for aneurysm repair, there is lesser availability of diseased aortic tissue for molecular, cellular, and histopathological analysis, therefore over the last three decades it has been proposed various small species models that have allowed the focal induction of these lesions for the study of physiopathological mechanisms and possible useful biomarkers as diagnostic and therapeutic targets. The present review article presents and discusses the animal models available as their applications, characteristics, advantages, and limitations for the development of preclinical studies, and their importance in the comprehension of this pathology in humans. [ABSTRACT FROM AUTHOR]

The study of antimicrobial compounds of animal origin, particularly antimicrobial peptides (AMPs), is a current research topic. However, extracting endogenous AMPs is a challenging process and requires the application of targeted enzymatic processing principles based on knowledge of the structure of prepropeptide molecules — precursors of AMPs. In this study, a search was conducted for antimicrobial peptides present in Sus scrofa and Bos taurus organisms, as well as their precursors, using The Antimicrobial Peptide Database and UniProtKB databases. In the amino acid sequences of prepropeptides, the sequences of the mature peptides were found, and cleavage sites for trypsin, bacterial collagenase (type I), and neutrophil elastase were determined. As a result of the search for antimicrobial compounds in The Antimicrobial Peptide Database, 18 antimicrobial peptides from Sus scrofa and 40 antimicrobial peptides from Bos taurus were identified. Based on the results of determining cleavage sites in AMP precursors, enzymes were ranked from less preferred to more preferred for AMP release as follows: bacterial collagenase (type I) ≤ trypsin < neutrophil elastase. This order is justified not only by the number of suitable cleavage sites and their accuracy but also by the action of enzymes within mature AMPs: it is important to consider that enzymes can “cut” the peptides themselves, thereby reducing their antimicrobial activity. The bioinformatics analysis conducted is applicable for both primary screening of raw material potential and determining of suitable enzymes for extracting antimicrobial compounds from Sus scrofa and Bos taurus organisms.

Abstract Neutrophil extracellular traps (NETs) are defense mechanisms that trap and kill microorganisms and degrade cytokines. However, excessive production, dysregulation of suppression mechanisms, or inefficient removal of NETs can contribute to increased inflammatory response and the development of pathological conditions. Therefore, research has focused on identifying drugs that inhibit or delay the NET release process. Since reactive oxygen species (ROS) play a significant role in NET release, we aimed to investigate whether resveratrol (RSV), with a wide range of biological and pharmacological properties, could modulate NET release in response to different stimuli. Thus, human neutrophils were pretreated with RSV and subsequently stimulated with PMA, LPS, IL-8, or Leishmania. Our findings revealed that RSV reduced the release of NETs in response to all tested stimuli. RSV decreased hydrogen peroxide levels in PMA- and LPS-stimulated neutrophils, inhibited myeloperoxidase activity, and altered the localization of neutrophil elastase. RSV inhibition of NET generation was not mediated through A2A or A2B adenosine receptors or PKA. Based on the observed effectiveness of RSV in inhibiting NET release, our study suggests that this flavonoid holds potential as a candidate for treating NETs involving pathologies.

Targeting neutrophil function has gained attention as a propitious therapeutic strategy for diverse inflammatory diseases. Accordingly, a series of enone-based derivatives were developed to inhibit neutrophil-mediated inflammation, showing promise for treating inflammatory diseases. These compounds fall into two clusters with distinct effects: one inhibits neutrophilic superoxide (SO) anion production and elastase release triggered by N-formyl-Met-Leu-Phe (fMLF), with compound 6a being most effective (IC50 values of 1.23 and 1.37 μM, respectively), affecting c-Jun N-terminal kinase (JNK) and Akt phosphorylation. The second cluster suppresses formation of SO anion without affecting elastase levels, surpassed by compound 26a (IC50 of 1.56 μM), which attenuates various mitogen-activated protein kinases (MAPKs) with minimal Akt impact. Notably, none of the tested compounds showed cytotoxicity in human neutrophils, underscoring their potential as therapeutic agents against inflammatory diseases.

AbstractThe recent global resurgence of severe infections caused by the Group A streptococcus (GAS) pathogen, Streptococcus pyogenes, has focused attention on this microbial pathogen, which produces an array of virulence factors, such as the pore-forming toxin, streptolysin O (SOT). Importantly, the interactions of SOT with human neutrophils (PMN), are not well understood. The current study was designed to investigate the effects of pretreatment of isolated human PMN with purified SOT on several pro-inflammatory activities, including generation of reactive oxygen species (ROS), degranulation (elastase release), influx of extracellular calcium (Ca2+) and release of extracellular DNA (NETosis), using chemiluminescence, spectrophotometric and fluorimetric procedures, respectively. Exposure of PMN to SOT alone caused modest production of ROS and elastase release, while pretreatment with the toxin caused significant augmentation of chemoattractant (fMLP)-activated ROS generation and release of elastase by activated PMN. These effects of treatment of PMN with SOT were associated with both a marked and sustained elevation of cytosolic Ca2+concentrations and significant increases in the concentrations of extracellular DNA, indicative of NETosis. The current study has identified a potential role for SOT in augmenting the Ca2+-dependent pro-inflammatory interactions of PMN, which, if operative in a clinical setting, may contribute to hyper-activation of PMN and GAS-mediated tissue injury.

Enzyme inhibition is one of the leading drug development methods for the treatment of many diseases. Due to the possible side effects and low bioavailability of existing drugs, studies are continuing for the discovery of new drugs. In this study, in vitro elastase, acetylcholinesterase inhibition activities of newly synthesized N,S-substituted polyhalogenated nitrobuta-1,3-dienes derivatives (compounds 3, 4a, 4b, 4c, 4d, 4e, and 4f), as well as their antioxidant properties, were investigated. Results was showed that compounds 4a (IC50 = 22.10 ± 0.49 µM), 4b (IC50 = 53.98 ± 1.77 µM), and 4f (IC50 = 32.01 ± 1.33 µM) showed higher elastase inhibition effect than positive control, ursolic acid (IC50 = 479.11 ± 15.53 µM) and in silico adsorption, distribution, metabolism, excretion, and toxicity (ADMET) and molecular docking studies were carried out in line with the results. As a result of molecular docking studies with iGemdock, DockThor, and Autodock Vina, it was determined that there was a higher binding relevance for compounds 4a (− 84.41/− 8.439 kcal/mol), 4b (− 86.32/− 7.878 kcal/mol), and 4f (− 86.32/− 8.530 kcal/mol) than ursolic acid (− 77.67/− 7.024 kcal/mol) for iGemdock and DockThor. It has been shown by DPPH, ABTS, and reducing power experiments that all compounds also show antioxidant properties. In conclusion, both in vitro and in silico molecular docking studies of compounds 4a, 4b, and 4f show that these three compounds are potent inhibitors of elastase. [ABSTRACT FROM AUTHOR]

Pulmonary vascular disease (PVD) complicated with pulmonary hypertension (PH) is a leading cause of mortality in congenital diaphragmatic hernia (CDH). Unfortunately, CDH patients are often resistant to PH therapy. Using the nitrogen CDH rat model, we previously demonstrated that CDH-associated PVD involves an induction of elastase and matrix metalloproteinase (MMP) activities, increased osteopontin and epidermal growth factor (EGF) levels, and enhanced smooth muscle cell (SMC) proliferation. Here, we aimed to determine whether the levels of the key members of this proteinase-induced pathway are also elevated in the pulmonary arteries (PAs) of CDH patients. Neutrophil elastase (NE), matrix metalloproteinase-2 (MMP-2), epidermal growth factor (EGF), tenascin-C, and osteopontin levels were assessed by immunohistochemistry in the PAs from the lungs of 11 CDH patients and 5 normal age-matched controls. Markers of proliferation (proliferating cell nuclear antigen (PCNA)) and apoptosis (cleaved (active) caspase-3) were also used. While expressed by both control and CDH lungs, the levels of NE, MMP-2, EGF, as well as tenascin-C and osteopontin were significantly increased in the PAs from CDH patients. The percentage of PCNA-positive PA SMCs were also enhanced, while those positive for caspase-3 were slightly decreased. These results suggest that increased elastase and MMPs, together with elevated tenascin-C and osteopontin levels in an EGF-rich environment may contribute to the PVD in CDH infants. The next step of this study is to expand our analysis to a larger cohort, and determine the potential of targeting this pathway for the treatment of CDH-associated PVD and PH. Therapeutic. LEVEL III. [ABSTRACT FROM AUTHOR]

BACKGROUND Abdominal aortic aneurysm (AAA) is an arterial disease characterized by dilatation of the aortic wall. It has been suggested that neutrophil counts and neutrophil elastase activity are associated with AAA. We investigated whether a neutrophil elastase (NE) inhibitor, sivelestat (Siv), had a protective effect against angiotensin II (AngII)-induced AAAs. METHODS Male apolipoprotein E-deficient mice were assigned into three groups: Vehicle + saline, AngII + saline, and AngII + Siv. All mice were administered intraperitoneally with either Siv or vehicle twice daily after AngII infusion. RESULTS In the 4-week AngII infusion study, plasma NE concentration (P  = 0.041) and its activity (P  = 0.011) were elevated by AngII. These increases were attenuated by Siv (concentration: P  = 0.010, activity: P  = 0.027). Further, plasma elastase activity was closely correlated with aortic width (R  = 0.6976, P  < 0.001). In the 1-week AngII infusion study, plasma and tissue elastase activity increased by AngII (plasma: P  = 0.034, tissue: P  < 0.001), but were reduced by Siv (plasma: P  = 0.014, tissue: P  = 0.024). AngII increased aortic width (P  = 0.011) but was attenuated by co-administration of Siv (P  = 0.022). Moreover, Siv decreased the incidence of AAAs (P  = 0.009). Elastin fragmentation induced by AngII was reduced by Siv. Many inflammatory cells that were either CD68 or Gr-1 positive were observed in the AngII + saline group, whereas few inflammatory cells were accumulated in the AngII + Siv group. MMP-2 and MMP-9 were enhanced by AngII, but were reduced by Siv. In vitro , MMP-2 activity was induced by human NE (medium: P  < 0.001, cells: P  = 0.001), which was attenuated by co-incubation of Siv in medium (P  < 0.001) and protein of human aortic smooth muscle cells (P  = 0.001). CONCLUSIONS Siv attenuated AngII-induced AAA through the inhibition of NE. [ABSTRACT FROM AUTHOR]

Objective: To investigate the digestive enzymes and biomarkers in the saliva of patients with laryngopharyngeal reflux (LPR) and asymptomatic individuals. Study Design: Prospective controlled study. Setting: Multicenter study. Methods: Patients with LPR at the hypopharyngeal‐esophageal impedance‐pH monitoring (HEMII‐pH) and asymptomatic individuals were consecutively recruited from January 2020 to April 2023 from 2 University Hospitals. The saliva of patients (off PPIs) and asymptomatic individuals was collected to measure pH, elastase, bile salts, cholesterol, gastric, and pancreatic lipases. Anxiety, symptoms, and findings were studied through perceived stress scale (PSS), reflux symptom score (RSS), and reflux sign assessment (RSA). Results: Sixty‐seven LPR patients and 57 asymptomatic individuals completed the evaluations. LPR patients reported higher PSS, RSS, and RSA than asymptomatic individuals. The mean saliva pH was more alkaline in LPR patients (7.23: 95% confidence interval [CI]: 7.08, 7.38) compared to controls (6.13; 95% CI: 5.95, 6.31; P =.001). The mean concentration of elastase was higher in patients (51.65 µg/mL; 95% CI: 44.47, 58.83 µg/mL) versus asymptomatic individuals (25.18 µg/mL; 95% CI: 21.64, 28.72 µg/mL; P =.001). The saliva cholesterol reported higher concentration in healthy individuals (3.43 mg/dL; 95% CI: 3.21, 3.65 mg/dL) compared to patients (1.16 mg/dL; 95% CI: 1.05, 1.27 mg/dL; P =.001). The saliva pH, and elastase concentration were significantly associated with the baseline RSS, while saliva cholesterol was negatively associated with the severity of RSS and RSA. Conclusion: Cholesterol, bile salts, and elastase are biomarkers of LPR and should be considered to develop future non‐invasive saliva device for the detection of LPR. [ABSTRACT FROM AUTHOR]

Neutrophil extracellular traps (NETs) are defense mechanisms that trap and kill microorganisms and degrade cytokines. However, excessive production, dysregulation of suppression mechanisms, or inefficient removal of NETs can contribute to increased inflammatory response and the development of pathological conditions. Therefore, research has focused on identifying drugs that inhibit or delay the NET release process. Since reactive oxygen species (ROS) play a significant role in NET release, we aimed to investigate whether resveratrol (RSV), with a wide range of biological and pharmacological properties, could modulate NET release in response to different stimuli. Thus, human neutrophils were pretreated with RSV and subsequently stimulated with PMA, LPS, IL-8, or Leishmania. Our findings revealed that RSV reduced the release of NETs in response to all tested stimuli. RSV decreased hydrogen peroxide levels in PMA- and LPS-stimulated neutrophils, inhibited myeloperoxidase activity, and altered the localization of neutrophil elastase. RSV inhibition of NET generation was not mediated through A2A or A2B adenosine receptors or PKA. Based on the observed effectiveness of RSV in inhibiting NET release, our study suggests that this flavonoid holds potential as a candidate for treating NETs involving pathologies. [ABSTRACT FROM AUTHOR]

Background: Pseudomonas Aeruginosa rarely causes disease in healthy people and can cause urinary tract infections(UTIs), bacteremia, eye infections, ear infections, skin infections, central nervous system infections, and bone and joint infections. Objectives : we aimed to study the virulence of genes :plc H, plc N and Las Bon isolates of these bacteria using PCR techniques. Materials and Methods : The study included the collection of (176) isolates from patients suffering from burns, wounds and urinary tract infections, where the samples were cultivated on different media, and the samples were diagnosed through cultural, morphological and microscopic characteristics. Results: The prevalence of this gene las B was 78.57% in burn isolates, 85.71% in wound isolates and 85.71% inurinary tract infection isolates .As for the presence of the gene plcN that encodes for the exogenous enzyme hemolysin,, wounds and urinary tract infection isolates. As for the presence of the gene plcH that encodes for the exogenous enzyme hemolysin, the rates were 85.71% in burn isolates. Conclusion : the detection of thegeneplcNthatfoundin92.85%in the burn, and the another gene was plcH and the last gene was las B that destroying the elastin protein and it has a lot to do with the virulence factor of P.areuginoso in the human’s blood. [ABSTRACT FROM AUTHOR]

The chronic inflammatory component of asthma is propagated by granulocytes, including neutrophils and eosinophils, in the peripheral circulation and airway. Previous studies have suggested that these cells have an altered expression of adhesion-related molecules and a propensity for the release of granule contents that may contribute to tissue damage and enhance inflammatory complications in patients with status asthmaticus. The goal of this prospective cohort study at a tertiary care pediatric hospital with a large population of asthma patients was to assess the role of granulocyte-based inflammation in the development of asthma exacerbation. Subjects were enrolled from two patient populations: those with mild-to-moderate asthma exacerbations seen in the emergency department and those with severe asthma admitted to the intensive care unit (PICU). Clinical data were collected, and blood was drawn. Granulocytes were immediately purified, and the phenotype was assessed, including the expression of cell surface markers, elastase release, and cytokine production. Severe asthmatics admitted to the PICU displayed a significantly higher total neutrophil count when compared with healthy donors. Moreover, little to no eosinophils were found in granulocyte preparations from severe asthmatics. Circulating neutrophils from severe asthmatics admitted to the PICU displayed significantly increased elastase release ex vivo when compared with the PMN from healthy donors. These data suggest that the neutrophil-based activation and release of inflammatory products displayed by severe asthmatics may contribute to the propagation of asthma exacerbations. [ABSTRACT FROM AUTHOR]

The carnivorous pitcher plants of the genus Nepenthes have long been known for their ethnobotanical applications. In this study, we prepared various extracts from the pitcher, stem, and leaf of Nepenthes miranda using 100% ethanol and assessed their inhibitory effects on key enzymes related to skin aging, including elastase, tyrosinase, and hyaluronidase. The cytotoxicity of the stem extract of N. miranda on H838 human lung carcinoma cells were also characterized by effects on cell survival, migration, proliferation, apoptosis induction, and DNA damage. The cytotoxic efficacy of the extract was enhanced when combined with the chemotherapeutic agent 5-fluorouracil (5-FU), indicating a synergistic effect. Flow cytometry analysis suggested that the stem extract might suppress H838 cell proliferation by inducing G2 cell cycle arrest, thereby inhibiting carcinoma cell proliferation. Gas chromatography–mass spectrometry (GC–MS) enabled the tentative identification of the 15 most abundant compounds in the stem extract of N. miranda. Notably, the extract showed a potent inhibition of the human RPA32 protein (huRPA32), critical for DNA replication, suggesting a novel mechanism for its anticancer action. Molecular docking studies further substantiated the interaction between the extract and huRPA32, highlighting bioactive compounds, especially the two most abundant constituents, stigmast-5-en-3-ol and plumbagin, as potential inhibitors of huRPA32's DNA-binding activity, offering promising avenues for cancer therapy. Overall, our findings position the stem extract of N. miranda as a promising source of natural compounds for anticancer therapeutics and anti-skin-aging treatments, warranting further investigation into its molecular mechanisms and potential clinical applications. [ABSTRACT FROM AUTHOR]

Objective: To explore the effect of neutrophil elastase (NE) on neutrophil inflammatory recruitment. Methods: Mice bone marrow-derived neutrophils were pretreated with an exogenous elastase inhibitor-sivelestat sodium. The effects of elastase inhibition on the in vivo inflammatory recruitment, chemotaxis, adhesion, cell polarization and spreading of NE were examined by peritonitis adoptive transfer assay, dunn chamber, flow chamber, immunofluorescence staining and spreading assay, respectively. The effects of elastase inhibition on NE phagocytosis and reactive oxygen species (ROS) release capacity were detected by flow cytometry and the luminol chemiluminescence system. Results: Sivelestat sodium pretreatment significantly attenuated neutrophil in vivo inflammatory recruitment (P<0.001); impaired neutrophil perception of chemotaxis in vitro (P<0.05), slowed chemotactic velocity (P< 0.05), and decreased the chemotactic distance (P<0.05); reduced the adhesion of neutrophils to inflamed endothelial cells (P<0.0001) and inhibited the phagocytosis of bacteria by neutrophils (P<0.01); however, there was no significant effect on neutrophil spreading, polarization and ROS. Conclusion: NE inhibition significantly impaired the inflammatory recruitment cascade response and phagocytosis of neutrophils in vitro and in vivo but had no significant effect on the spreading, polarization and ROS release of neutrophils. [ABSTRACT FROM AUTHOR]

We have previously shown that Bacillus sp. IMV B-7883 exhibits both elastase and fibrinogenolytic activity. One of the approaches to enhance enzymatic activity is the use of coordination compounds capable to affect enzyme’s activity or synthesis. The purpose of this work was to study the effect of mixed-ligand complexes of Ge(IV) – Co(II) (Ni(II), Cu(II)) with 1-hydroxyethane-1,1-diphosphonic acid аnd 2,2′-bipyridine on the activity of elastase and fibrinogenase purified from Bacillus sp. IMV B-7883. Previously synthesized and characterized mixed-ligand complexes and enzymes purified from the supernatant of the bacterial culture liquid were used in the study. Elastase activity was determined colorimetrically with the use of Congo red, fibrinogenase activity was estimated by fibrinogen hydrolysis measured by absorption at 275 nm. It was shown that complexes 1 (C132H164Co4Ge6N24O68P12) and 2 (C132H148Ge6N24Ni4O60P12) inhibited activity of Bacillus sp. IMV B-7883 elastase by 54 and 71% respectively, while complex 3 (C92H128Cu4Ge6N16O63P12) enhanced it by 30%. Stimulating effect of all three complexes on fibrinogenase activity was revealed. Thus, complex 1 and 2 activated the enzyme by more than 50% and complex 1 – by 19%. The data obtained indicate a complex mechanism of the studied complexes influence on enzymatic activity depending on both their composition and structure.

Chios mastic gum (CMG), the resin of Pistacia lentiscus var. Chia, is a product with great ethnopharmacological and economic significance. This study attempts to investigate, for the first time, the activity of CMG, its fractions and isolated compounds against specific enzymes, which play pivotal roles in the degradation of proteins contained in skin connective tissue. Initially, crude CMG was subjected to extraction, fractionation and isolation through different chromatographic techniques to obtain the acidic and neutral fraction of terpenes. Additionally, the characteristic and major active triterpene acids of CMG, masticadienonic and isomasticadienonic acids (MNA, IMNA) were isolated in pure form. All samples were analysed by means of High-Performance Thin-Layer Chromatography (HPTLC) with four distinct development systems to obtain their constituents’ profile. Finally, samples were tested for their ability to inhibit the elastase and collagenase enzymes. According to our findings, for collagenase, a mixture of MNA and IMNA demonstrated the most potent activity with an IC50 value of 31.07 μg/mL, while for elastase CMG’s acidic fraction provided the most promising results with an IC50 value of 17.30 μg/mL. Overall, these results attempt to fill the gap in scientific knowledge about the use of CMG and its constituents in skincare and cosmetic products.

Emphysema is one of the major components of chronic obstructive pulmonary disease (COPD), which is characterised by the destruction and enlargement of air spaces, leading to airflow limitation and dyspnoea, finally progressing to oxygen dependency. The alveolar wall destruction is due to chronic inflammation, oxidative stress, apoptosis, and proteinase/anti-proteinase imbalance. So far, there has been no effective therapy for patients with COPD. We evaluated the therapeutic efficacy of tannic acid (TA), a naturally occurring plant-derived polyphenol in the murine emphysema model. In C57BL/6 J mice, we established emphysema by intratracheal instillation of elastase (EL). Then, mice were treated with TA and evaluated 1 and 21 days post-EL instillation. After 24 h, TA treatment significantly reduced EL-induced histopathological alterations, infiltrating leukocytes, and gene expression of markers of inflammation and apoptosis. Similarly, after 21 days, TA treatment suppressed the mean linear intercept, gene expression of proteinases, and increased elastic fiber contents in the lungs when compared to the EL-alone group. Furthermore, EL induced the activation of p38 mitogen-activated protein kinase (MAPK) and nuclear factor kappa light chain enhancer of activated B cells (NF-kB) p65 pathways in the lungs was suppressed by TA treatment. In summary, TA has the potential to mitigate EL-induced inflammation, apoptosis, proteinase/anti-proteinase imbalance, and subsequent emphysema in mice. [ABSTRACT FROM AUTHOR]

Thoracic aortic aneurysm (TAA) has a prevalence of 0.16–0.34% and an incidence of 7.6 per 100,000 person-years, accounting for 1–2% of all deaths in Western countries. Currently, no effective pharmacological therapies have been identified to slow TAA development and prevent TAA rupture. Large TAAs are treated with open surgical repair and less invasive thoracic endovascular aortic repair, both of which have high perioperative mortality risk. Therefore, there is an urgent medical need to identify the cellular and molecular mechanisms underlying TAA development and rupture to develop new therapies. In this review, we summarize animal TAA models including recent developments in porcine and zebrafish models: porcine models can assess new therapeutic devices or intervention strategies in a large mammal and zebrafish models can employ large-scale small-molecule suppressor screening in microwells. The second part of the review covers current views of TAA pathogenesis, derived from recent studies using these animal models, with a focus on the roles of the transforming growth factor-beta (TGFβ) pathway and the vascular smooth muscle cell (VSMC)-elastin-contractile unit. The last part discusses TAA treatment options as they emerge from recent preclinical studies. [ABSTRACT FROM AUTHOR]

Encapsulating drugs in poly(lactic‐co‐glycolic acid) (PLGA) nanoparticles can provide a gradual and consistent drug release. Herein, diclofenac sodium‐encapsulated PLGA nanoparticles (DS‐PLGA NPs) were synthesized, characterized, and their effects on bacterial proteolysis and elastolytic activity were evaluated using skim milk agar plate, elastin‐congo red, and staphylolysin disk susceptibility assays. Quantitative PCR was employed to examine the effect of nanoparticles on the expression of lasA/B genes. The DS‐PLGA NPs displayed spherical morphology with an average size of 86 nm, zeta potential of ‐15.2 mV, and DLS size of 124.6 nm. At a sub‐inhibitory concentration, DS‐PLGA NPs inhibited bacterial elastolytic activity by 41.6 to 62.1% during 30–180 min of incubation. The elastolytic curve slopes of samples from PLGA and DS‐PLGA treated groups were respectively 0.0714 and 0.0380 activity/min, indicating a significant reduction of bacterial extracellular elastases by DS‐PLGA NPs. Exposure of bacteria to a sub‐inhibitory concentration of the nanoparticles attenuated the expression of the lasA and lasB genes by 0.21 and 0.31 folds, respectively. Attenuation of the proteolytic activity of P. aeruginosa suggests that the antivirulence potential of the DS‐PLGA NPs along with the anti‐inflammatory properties of diclofenac sodium, can be considered for the treatment of acute and chronic P. aeruginosa infections. [ABSTRACT FROM AUTHOR]

Objective To determine the time point when porcine pancreatic elastase(PPE)induced abdominal aortic aneurysm(AAA)reaches the regression phase in mice and observe the histological characteristics of AAA in regression phase. Methods AAAs were induced by transient intraluminal infusion of PPE in C57BL/6J mice. The diameters of the mouse abdominal aortas were measured before PPE infusion and sacrifice time, day 14 for AAA progression phase or day 56 for regression phase after PPE infusion, respectively. The histological characteristics of the aneurysm lesion site on day 14 and day 56 after surgery were compared and analyzed. Results The diameters of the abdominal aortas were significantly increased in both day 14 and day 56 after PPE infusion groups(diameter growth rate 147% and 155%, respectively)as compared to the baseline diameters. In the day 14 group, the infused aortas showed typical AAA characteristics, such as elastin break/degradation, medial smooth muscle cells depletion, and inflammatory cell diffused infiltration. In the day 56 group after PPE infusion, although the artery diameter did not change significantly as compared to the day 14 group, histology showed that elastin was partially repaired, new smooth muscle cells were added to the damaged aorta media, the infiltrated inflammatory cells were significantly subsided, and the adventitia neovascularization was reduced, showing a significant feature of the disease regression phase. Conclusion In the PPE-induced mouse AAA model, day 56 after surgery is an appropriate time point for observing aneurysm regression, and the histological characteristics of the regression are obvious. [ABSTRACT FROM AUTHOR]

Conventional models view b1-adrenergic receptors (b1ARs) as full-length proteins that activate signaling pathways that influence contractile function and ventricular remodeling - and are susceptible to agonist-dependent desensitization. This perspective summarizes recent studies from my laboratory showing that post-translational processing of the b1-adrenergic receptor N-terminus results in the accumulation of both full-length and Nterminally truncated forms of the b1AR that differ in their signaling properties. We also implicate oxidative stress and b1AR cleavage by elastase as two novel mechanisms that would (in the setting of cardiac injury or inflammation) lead to altered or decreased b1AR responsiveness [ABSTRACT FROM AUTHOR]

Simple Summary: The early detection of pancreatic cancer is critical as it is usually too late for potentially curative surgical resection when obvious symptoms such as jaundice have developed. The pancreas has two main functions in the body, namely, an endocrine role, where it produces insulin to control blood glucose levels; and an exocrine role, where it produces enzymes that aid digestion. Impairment of this latter role is known as pancreatic exocrine insufficiency (PEI). It is an established feature of advanced pancreatic cancer, but it is unclear whether it is present in the early stages when surgery may still be an option. This study used two validated methods of testing pancreatic exocrine function, a breath test and a stool test, to demonstrate that exocrine insufficiency is associated with resectable pancreatic cancer. Pancreatic ductal adenocarcinoma (PDAC) is usually diagnosed late, leading to a high mortality rate. Early detection facilitates better treatment options. The aim of this UK-based case–control study was to determine whether two validated tests for pancreatic exocrine insufficiency (PEI), namely, the 13C-mixed triglyceride breath test (13C-MTGBT) and a faecal elastase (FE-1) test, can discriminate between patients with resectable PDAC versus healthy volunteers (HVs) along with a comparison group with chronic pancreatitis (CP). Discrimination between disease states and HVs was tested with receiver operator characteristic (ROC) curves. In total, 59 participants (23 PDAC (16 men), 24 HVs (13 men) and 12 CP (10 men)) were recruited, with a similar age in each population, and a combined median (IQR) age of 66 (57–71). The areas under the ROC curve for discriminating between PDAC and HVs were 0.83 (95% CI: 0.70–0.96) for the 13C-MTGBT, and 0.85 (95% CI: 0.75–0.95) for the FE-1 test. These were similar to CP vs. HV. In conclusion, PEI occurs in resectable PDAC to a similar extent as in CP; further large-scale, prospective studies using these tests in the primary care setting on high-risk groups are warranted. [ABSTRACT FROM AUTHOR]

The Eucalyptus tree, belonging to the myrtle family, grows all over the world for its pharmaceutical and industrial benefits. In this article, we present a comparative analysis of the chemical composition of the hydrodistilled oils obtained from three different Eucalyptus species growing in Egypt viz. E. citriodora, E. camaldulensis, and E. ficifolia. Gas Chromatography-Mass Spectrometric guided analysis resulted in the identification of a total of 20 metabolites in E. citriodora oil with citronellal (54.9%) and citronellol (25.4%) being the most dominant components. β-cymene (12.7%) and 1,8-cineole (11.7%) were the major volatile constituents identified in E. camaldulensis oil, while trans-β-ocimene (22.4%), 1,8-cineole (13.5%), and L-trans-pinocarveol (12.5%) were the dominating components in the oil of E. ficifolia. The essential oils of the studied species were evaluated for their in vitro anti-inflammatory, antiviral including anti-SARS-CoV-2 (severe acute respiratory syndrome corona virus 2), antibacterial, and antifungal activities. E. citriodora oil displayed the highest inhibitory activity on the release of the superoxide radical (32%) and elastase enzyme (31%) in human neutrophils, while E. ficifolia oil had enhancing effects on elastase. The latter showed significant antiviral effects against hepatitis A, herpes simplex, and coxsackie viruses with IC50 values at 2.1, 2.5, and 5.6 μg/mL, respectively. Moderate antibacterial and antifungal activities were observed for Eucalyptus oils with Staphylococcus aureus being the most susceptible bacterial strain. E. ficifolia oil, similarly, displayed the best antibacterial activity with minimum inhibitory concentration (MIC) value at ca. 25 μg/mL (for S. aureus). On the contrary, E. camaldulensis oil was the most active against Candida albicans with an MIC value at 45 μg/mL. In silico studies were performed with a number of macromolecular drug targets for confirming the biological activities of the identified compounds and for interpreting their ADME (absorption-distributionmetabolism-elimination) parameters. [ABSTRACT FROM AUTHOR]

Introduction: Neutrophil extracellular traps (NETs) are structures released by neutrophils in response to various infections. NETs have a biocidal role and have been demonstrated to be effective against bacteria, fungi, viruses, and parasites. Depending on the situation, NETs can protect the host from pathogen invasion or contribute to the development of autoimmune diseases such as cystic fibrosis and rheumatoid arthritis. In this study, we aimed to investigate the occurrence of NET as one of the components in upper respiratory tract secretions in infectious and allergic diseases. Methods: Nasal mucus was collected from donors diagnosed with infectious rhinitis or allergic rhinitis. The extracellular DNA content was determined using SytoxGreen staining, and the total protein pool was determined using the microBCA method. Micrococcal nuclease was used to digest the samples and ELISA was employed to identify the NET proteins. The enzymatic activity of elastase was determined. Results: Our findings showed that nasal mucus collected from patients with infectious rhinosinusitis contained extracellular DNA that could come from a variety of sources, responsible for increasing the density and viscosity of secretions, as well as NETs proteins. The identified enzymatic activity of NET elastase indicates the possible irritation of nasal tissues. However, the DNA content was not identified in the samples from allergic patients. In addition, we have shown in preliminary studies that therapy using N-acetylcysteine can liquefy nasal secretions. Discussion: The study suggests that the composition of nasal mucus varies according to the cause of mucosal irritation. The presence of DNA and NET proteins can have severe consequences for the therapeutic process prolonging treatment. The low viscosity of nasal mucus in allergic patients facilitates mucosal flushing and the removal of allergens. Understanding the occurrence and role of NETs in various respiratory diseases is critical for developing effective treatment strategies that consider the complex interaction between the immune system and pathogens. The results of this study suggest that NETs may be present in upper respiratory tract secretions with an infectious background, supporting basic defense mechanisms using eosinophils and EETs. Further research is needed to explore the potential of NETs as a therapeutic target in respiratory diseases. [ABSTRACT FROM AUTHOR]

Callistemon is an aromatic genus of flowering plants belonging to family Myrtaceae. The essential oils of C. subulatus leaves were collected in four seasons and analyzed using GC/MS. The oils demonstrated monoterpenes as the predominant class. Eucalyptol was the main component in all seasons; summer (66.87%), autumn (58.33%), winter (46.74%) and spring (44.63%), followed by α-pinene; spring (31.41%), winter (28.69%), summer (26.34%) and autumn (24.68%). Winter oil, the highest yield (0.53 mL/100g), was further investigated for its inhibitory activity against enzymes associated with ageing; elastase and acetylcholinesterase. It remarkably inhibited elastase and acetylcholinesterase with IC50 values of 1.05 and 0.20 µg/ml, respectively. A molecular docking study was conducted for the major oil components on the active sites of target enzymes. Eucalyptol revealed the best binding affinity for both enzymes. C. subualtus oil could be used as supplement for management of ageing disorders like skin wrinkles and dementia. [ABSTRACT FROM AUTHOR]

The study reports the biochemical characterization and mechanism of action of a novel 19.6 kDa protease inhibitor (PIs) isolated from the seeds of Caesalpinia decapetala belonging to the Fabaceae family. A systematic study was performed to ascertain the purity, specificity, biochemical and structural characterization, and its potential in curbing inflammation in vitro conditions. A two‐step chromatography technique was used to purify the PIs. Sodium dodecyl sulfate polyacrylamide gel electrophoresis and matrix‐assisted laser desorption ionization time of flight were employed to detect the molecular mass of the protein. N‐terminal sequence analysis of the inhibitor showed sequence similarity with the Kunitz family PIs. The in vitro test tube assay was performed for determining the anti‐inflammatory activity and the inhibitor is antiproliferative against macrophage (RAW264.7) and lung cancer cell lines (A549). An effective decrease in the release of inflammatory mediators (NO, IL‐6, TNF‐α) and on the activity of elastase was observed in macrophage cell lines (RAW264.7) which were treated with PIs. The purified inhibitor shows promising results against inflammation. Significance statement: Protease and antiprotease imbalance have an impact on several biological systems which are reported to cause emphysema, thrombosis, clotting disorders, and cancer. Our findings clearly reveal that purified preatease inhibitor, CdTI have significant action as an anti‐proinflammatory and antiproliferative action on lung cancer cells. These findings could lead to protease inhibitors (PIs) being classified as therapeutic potential due to antiproliferative activity on cancer cell lines. These studies provide evidence about the therapeutic potential of serine PIs in inhibiting or regulating the activity of serine proteases secreted by neutrophils. [ABSTRACT FROM AUTHOR]

Ovine footrot is a contagious bacterial disease that causes foot lesions, and depending on the virulence of the causative strains, may lead to severe underrunning of the hoof and lameness. Virulent footrot can be identified, treated and controlled more effectively than less virulent benign forms. The in vitro elastase test for virulence of the causative bacteria, Dichelobacter nodosus, has been used to support clinical diagnosis. However, not all laboratory‐designated virulent D. nodosus strains cause clinical signs of virulent footrot. This study evaluated retrospectively how well the elastase test supported clinical footrot diagnosis in 150 sheep flocks examined for suspect footrot in New South Wales between August 2020 and December 2021. Flocks were included if measures of clinical disease, environmental conditions and the virulence of D. nodosus isolates were available. Variation in the elastase activity result between D. nodosus isolated from the same flock made bacterial virulence hard to interpret, but calculating the mean elastase rate for all isolates from the same flock made correlations between bacterial virulence and flock footrot diagnosis possible. Simplifying bacterial virulence into whether there were any elastase‐positive D. nodosus isolates before 12 days increased the predictive value of elastase results for virulent diagnosis, compared with using the first day that any isolate was elastase positive or the percentage of elastase‐positive isolates by 12 days, but not all clinically virulent flocks had isolates with elastase activity before 12 days. Logistic regression models were fitted to identify the minimum number of predictors for virulent footrot diagnosis, with models suggesting that virulent footrot diagnosis was best predicted by adding the elastase test result and environmental conditions to the prevalence of severe foot lesions (score 4 and 5). However, performing the same analysis with different breeds, ages of sheep and seasons might highlight other factors important in the diagnosis of virulent footrot. [ABSTRACT FROM AUTHOR]

Natural skin aging is indicated by a loss of skin structure and integrity caused by external factors including UV exposure. This exposure causes oxidative stress on skin cells, initiates aging, and degradation of extracellular matrix (ECM) structure which is composed of many proteins, including collagen and elastin. ECM degradation is caused by the increased activity of proteolytic enzymes, one of which is elastase (closely related to wrinkling). To inhibit oxidative stress during the photoaging process, the skin needs antioxidant compounds. Salak (Salacca zalacca (Gaert.)Voss) is a fruit that is rich in antioxidants because it contains flavonoid, phenolic, and polyphenolic compounds, including chlorogenic acid. The purpose of this study is to demonstrate the antioxidant activity and antiaging properties of salak fruit extract (SFE) as a candidate for active ingredients in the prevention of aging. To determine the antioxidant activity of SFE, 2,2-diphenyl-1-picrylhydrazil (DPPH) scavenging and ferric reducing antioxidant power (FRAP) assays were performed, while elastase assays were performed to determine the anti-aging properties of SFE. The IC50 values for DPPH scavenging and FRAP activity of SFE were 107.52 μg/mL and 16.82 μg/mL with the highest activity at 68.79% and 97.96%, at concentrations of 200 and 50 μg/mL, respectively. Meanwhile, the anti-aging properties determined through the elastase assay showed an IC50 value of 19.71 μg/mL with the highest inhibition 72.50% at a concentration of 66.67 μg/mL. SFE has the potential as an active ingredient in preventing aging through its antioxidant activity and anti-aging properties.

The representatives of Bacillus are some of the best protease producers studied so far since they exhibit broad substrate specificity, significant activity, stability, simple downstream purification, short period of fermentation and low cost. Earlier, we showed that Bacillus sp. IMV B-7883 strain synthesizes an extracellular proteases, which exhibit elastolytic and fibrinogenolytic activity. The aim of the work was to isolate and purify these enzymes from the culture liquid of the Bacillus sp. IMV B-7883 strain, as well as to study their properties. Isolation and purification of proteases was carried out by precipitation of the culture liquid with ammonium sulfate, gel permeation and ion exchange chromatography and rechromatography on Sepharose 6B. As a result, proteases with elastolytic and fibrinogenolytic activity with a molecular weight of 23 and 20 kDa respectively were isolated with elastase activity increased by 63.6 and fibrinogenolytic activity by 44.1 times. The enzyme with elastase activity had a pH-optimum of 7.0 and hydrolyzed only elastin, while the enzyme with fibrinogenolytic activity was an alkaline protease with a pH-optimum of 8.0 and in addition to fibrinogen, showed specificity for fibrin and, in trace amounts, for collagen.

Abstract A new compound, C23H20BrN3OS, containing a quinoline-based iminothiazoline with a thiazoline ring, was synthesized and its crystal and molecular structures were analyzed through single crystal X-ray analysis. The compound belongs to the triclinic system P − 1 space group, with dimensions of a = 9.2304 (6) Å, b = 11.1780 (8) Å, c = 11.3006 (6) Å, α = 107.146 (5)°, β = 93.701 (5)°, γ = 110.435 (6)°, Z = 2 and V = 1025.61 (12) Å3. The crystal structure showed that C–H···N and C–H···O hydrogen bond linkages, forming infinite double chains along the b-axis direction, and enclosing R2 2(14) and R2 2(16) ring motifs. The Hirshfeld surface analysis revealed that H…H (44.1%) and H…C/C…H (15.3%) interactions made the most significant contribution. The newly synthesized (Z)-4-bromo-N-(4-butyl-3 (quinolin-3-yl)thiazol-2(3H)-ylidene)benzamide, in comparison to oleanolic acid, exhibited more strong potential against elastase with an inhibition value of 1.21 µM. Additionally, the derivative was evaluated using molecular docking and molecular dynamics simulation studies, which showed that the quinoline based iminothiazoline derivative has the potential to be a novel inhibitor of elastase enzyme. Both theoretical and experimental findings suggested that this compound could have a number of biological activities.

Context: The leading cause of skin sunburn is exposure to ultraviolet radiation (UVR), which can also cause reactive oxygen species (ROS), which accelerates aging. Rubus fraxinifolius Poir. leaf ethanolic extract may have antiaging properties by preventing elastase enzyme activity. Aims: To compare the antioxidant activity and elastase enzyme inhibition from old and young leaves using Soxhlet extraction and evaluate the antioxidant activity and elastase enzyme inhibition utilizing a variety of extraction techniques, including conventional Soxhlet extraction and unconventional methods like microwave-assisted extraction (MAE) and ultrasonic-assisted extraction (UAE). Methods: Using various extraction methods, samples were extracted with 96% ethanol (Soxhlet, MAE, and UAE). The active extract was then transformed into n-hexane, ethyl acetate, and distilled water fraction. Following the total phenolic and total flavonoid content analysis, the antioxidant activity was evaluated using the radical scavenging tests of 1,2-diphenyl-2-picrylhydrazyl (DPPH), ferric reducing antioxidant power assay (FRAP), and ABTS radicals, or 2,2′-azino-bisphenol S (2,2′-ABPS) (3-ethylbenzothiazoline-6-sulphonic acid). The antiaging potential of skin breakdown enzymes was examined using elastase. Results: According to the R. fraxinifolius leaf’s optimization type, extract from older leaves performs better than from younger leaves in terms of antioxidant activity and elastase enzyme inhibitory activity. In terms of optimization extraction, UAE approaches to produce the best outcomes than Soxhlet and MAE. Additionally, using the chosen UAE extraction method, multilayer fractionation was done using n-hexane, ethyl acetate, and distilled water solvents, with distilled water fraction producing the best results. Conclusions: The distilled water fraction from R. fraxinifolius leaf’s shows the potential for future research to support its efficacy as an active component in antiaging cosmetics.

Type 2 diabetes mellitus (T2DM) is a metabolic syndrome known to contribute to impaired wound healing. This condition can be further worsened by excessive melanin production, elastin degradation, and chronic infections at the wound site, potentially leading to melasma and diabetic dermopathy. The purpose of this study was to investigate the phytochemical profile and inhibitory effects of Tetraclinis articulata essential oil (TAEO) on target enzymes involved in diabetes pathogenesis and chronic wound remodeling, namely α-amylase, α-glucosidase, tyrosinase, and elastase, as well as its in vitro antibacterial activity. Gas chromatography and mass spectrometry (GC-MS) analysis of TAEO led to the identification of 46 volatile compounds, representing 96.61 % of TAEO. The major metabolites were bornyl acetate (29.48 %), α-pinene (8.96 %), germacrene D (7.70 %), and d-limonene (5.90 %). TAEO exhibited limited scavenging activity against DPPH free radicals, whereas the FRAP and ABTS assays indicated a relatively higher antioxidant activity. Remarkably, TAEO disclosed a promising in vitro antidiabetic activity against α-glucosidase with an IC50 value of 178 ± 1.6 μg/mL, which is comparable to the standard inhibitor acarbose (IC50 = 143 ± 1.1 μg/mL). In silico, molecular docking analysis against α-glucosidase identified 15 compounds that interacted with the enzyme's active site, whereas skin permeability and sensitization assessments indicated that 26 out of the 44 identified volatile compounds were predicted to be free from any skin sensitivity risk. On the other hand, moderate inhibitory activity was recorded against α-amylase, tyrosinase, and elastase. Notably, TAEO at 5 % significantly suppressed biofilm formation by P. aeruginosa, S. aureus, and E. faecalis, common skin pathogens associated with wound infections, and reduced their swarming motility. Our findings suggest that TAEO may hold the potential as a natural remedy for type 2 diabetes and its associated co-morbidities, especially chronic wounds.

Neutrophils have a critical role in the innate immune response to infection and the control of inflammation. A key component of this process is the release of neutrophil serine proteases (NSPs), primarily neutrophil elastase, proteinase 3, cathepsin G, and NSP4, which have essential functions in immune modulation and tissue repair following injury. Normally, NSP activity is controlled and modulated by endogenous antiproteases. However, disruption of this homeostatic relationship can cause diseases in which neutrophilic inflammation is central to the pathology, such as chronic obstructive pulmonary disease (COPD), alpha-1 antitrypsin deficiency, bronchiectasis, and cystic fibrosis, as well as many non-pulmonary pathologies. Although the pathobiology of these diseases varies, evidence indicates that excessive NSP activity is common and a principal mediator of tissue damage and clinical decline. NSPs are synthesized as inactive zymogens and activated primarily by the ubiquitous enzyme dipeptidyl peptidase 1, also known as cathepsin C. Preclinical data confirm that inactivation of this protease reduces activation of NSPs. Thus, pharmacological inhibition of dipeptidyl peptidase 1 potentially reduces the contribution of aberrant NSP activity to the severity and/or progression of multiple inflammatory diseases. Initial clinical data support this view. Ongoing research continues to explore the role of NSP activation by dipeptidyl peptidase 1 in different disease states and the potential clinical benefits of dipeptidyl peptidase 1 inhibition.