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The article discusses a case study of a 26-year-old female patient with anetoderma, initially presenting as leprosy due to prolonged steroid use. The patient had asymptomatic oval and saccular papules and plaques on her upper limbs, with histopathological evidence confirming indeterminate leprosy. The study highlights the challenges of diagnosing leprosy in patients with prolonged steroid use and emphasizes the importance of a comprehensive diagnostic approach and pharmacovigilance in endemic areas. [Extracted from the article]

• 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]

The significance of Thymus species in the scientific community is growing steadily due to their extensive utilization in traditional medicine, food industry, and pharmaceutical sector, owing to their abundance in essential oil and phytochemical content, rendering them commercially significant species. The current work focuses on conducting a comprehensive analysis of the ethanol extract and essential oil derived from the root and aerial portions of Thymus brachychilus Jalas, an endemic species that has not been previously investigated. Additionally, a novel GC-MS (gas chromatography–mass spectrometry) technique was developed to quantify the levels of triterpenoids, which are frequently found in many plants, particularly those belonging to the Lamiaceae family. The approach was then used to assess the triterpenoid content of the species. While the phenolic content of the species was determined by LC-MS/MS (liquid chromatography–tandem mass spectrometry), the chemical composition of triterpenoid, essential oil and flavor (aroma) of the plant was determined by GC-MS. Eucalyptol was the primary ingredient in both the essential oil and the flavor, accounting for 11.05% and 12.35%, respectively. In the DPPH and ABTS radical scavenging techniques, the root ethanol extract exhibited the greatest antioxidant activity with IC50 values of 26.70 ± 0.23 and 19.16 ± 0.11 µg/mL, respectively. There was a high level of urease (59.54 ± 1.67% at 100 µg/mL) inhibitory activity observed in the root ethanol extract in enzyme inhibition assays, as well as angiotensin (94.80 ± 0.56%), elastase (40.19 ± 0.39%), and collagenase (48.26 ± 0.12%) inhibition in the aerial ethanol extract. Moreover, the MCF-7 cell line for breast cancer had a strong cytotoxic impact when exposed to the essential oil of the species (vitality%: 1.45 ± 0.01 at 200 µg/mL. The LC-MS/MS and GC-MS studies revealed that the roots had a significant concentration of rosmarinic acid (15,801 µg analyte/g extract), but the aerial portions had a substantial quantity of ursolic acid (152,374 µg analyte/g extract). The plant exhibits potential in the food, cosmetic, and pharmaceutical sectors on account of its elevated levels of rosmarinic and ursolic acid, superior antioxidant capacity, and angiotensin and urease inhibitory effect. [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]

Acute inflammation has the potential for the recruitment of immune cells, inhibiting tumor angiogenesis, metastasis, and drug resistance thereby overcoming the tumor immunosuppressive microenvironment caused by chronic inflammation. Here, an acute inflammation inducer using bacteria outer membrane vesicles (OMVs) loaded in thermal‐sensitive hydrogel (named OMVs‐gel) for localized and controlled release of OMVs in tumor sites is proposed. OMVs trigger neutrophil recruitment and amplify acute inflammation inside tumor tissues. The hydrogel ensures drastic inflammation is confined within the tumor, addressing biosafety concerns that the direct administration of free OMVs may cause fatal effects. This strategy eradicated solid tumors safely and rapidly. The study further elucidates one of the possible immune mechanisms of OMVs‐gel therapy, which involves the assembly of antitumor neutrophils and elastase release for selective tumor killing. Additionally, tumor vascular destruction induced by OMVs‐gel results in tumor darkening, allowing for combinational photothermal therapy. The findings suggest that the use of OMVs‐gel can safely induce acute inflammation and enhance antitumor immunity, representing a promising strategy to promote acute inflammation application in tumor immunotherapy. [ABSTRACT FROM AUTHOR]

The increase in the number of hospital strains of hypervirulent and multidrug resistant (MDR) Pseudomonas aeruginosa is a major health problem that reduces medical treatment options and increases mortality. The molecular profiles of virulence and multidrug resistance of P. aeruginosa-associated hospital and community infections in Mexico have been poorly studied. In this study, we analyzed the different molecular profiles associated with the virulence genotypes related to multidrug resistance and the genotypes of multidrug efflux pumps (mex) in P. aeruginosa causing clinically critical infections isolated from Mexican patients with community- and hospital-acquired infections. Susceptibility to 12 antibiotics was determined using the Kirby–Bauer method. The identification of P. aeruginosa and the detection of virulence and efflux pump system genes were performed using conventional PCR. All strains isolated from patients with hospital-acquired (n = 67) and community-acquired infections (n = 57) were multidrug resistant, mainly to beta-lactams (ampicillin [96.7%], carbenicillin [98.3%], cefalotin [97.5%], and cefotaxime [87%]), quinolones (norfloxacin [78.2%]), phenicols (chloramphenicol [91.9%]), nitrofurans (nitrofurantoin [70.9%]), aminoglycosides (gentamicin [75%]), and sulfonamide/trimethoprim (96.7%). Most strains (95.5%) isolated from patients with hospital- and community-acquired infections carried the adhesion (pilA) and biofilm formation (ndvB) genes. Outer membrane proteins (oprI and oprL) were present in 100% of cases, elastases (lasA and lasB) in 100% and 98.3%, respectively, alkaline protease (apr) and alginate (algD) in 99.1% and 97.5%, respectively, and chaperone (groEL) and epoxide hydrolase (cif) in 100% and 97.5%, respectively. Overall, 99.1% of the strains isolated from patients with hospital- and community-acquired infections carried the efflux pump system genes mexB and mexY, while 98.3% of the strains carried mexF and mexZ. These findings show a wide distribution of the virulome related to the genotypic and phenotypic profiles of antibiotic resistance and the origin of the strains isolated from patients with hospital- and community-acquired infections, demonstrating that these molecular mechanisms may play an important role in high-pathogenicity infections caused by P. aeruginosa. [ABSTRACT FROM AUTHOR]

Aims Methods Results Conclusion Bronchiectasis patients face an unmet need for treatment options that reduce inflammation. Cathepsin C inhibition is expected to achieve this by reducing the activation of neutrophil‐derived serine proteases. Here, we present safety and pharmacokinetic (PK) data from a Phase I trial evaluating the novel cathepsin C inhibitor BI 1291583 in healthy Japanese male subjects.This randomized, double‐blind, placebo‐controlled, parallel‐group study investigated BI 1291583 in healthy Japanese male subjects (jRCT2071210111) and consisted of a single‐rising‐dose (SRD) part and a multiple‐dose (MD) part. The primary endpoint was the percentage of subjects with drug‐related treatment‐emergent adverse events (AEs). Secondary PK endpoints (SRD: AUC0‐∞ and Cmax; MD: AUCτ,1 and Cmax,1 after first dose and AUCτ,ss and Cmax,ss after last dose), as well as further safety and PK endpoints, were also assessed.Overall, 36 subjects (n = 24 for SRD part; n = 12 for MD part) entered this Phase I trial. BI 1291583 was safe and well tolerated across the doses tested. All AEs were of mild intensity, with no drug‐related treatment‐emergent AEs, deaths, serious AEs or AEs of special interest reported in either part of the trial. Following both SRD and MD administration, BI 1291583 was readily absorbed, and PK was supraproportional over the doses assessed.The results show that BI 1291583 has an appropriate benefit–risk ratio for Japanese patients, with no safety or exposure concerns at the doses studied. Japanese patients with bronchiectasis can be safely integrated into future global clinical trials of BI 1291583, with no dose adjustment required. [ABSTRACT FROM AUTHOR]

Whereas aGVHD has strong inflammatory components, cGVHD displays autoimmune and fibrotic features; incidence and risk factors are similar but not identical; indeed, the aGVHD is the main risk factor for cGVHD. Calcineurin Inhibitors (CNI) with either Methotrexate (MTX) or Mycophenolate (MMF) still represent the standard prophylaxis in HLA-matched allogeneic stem cell transplantation (HSCT); other strategies focused on ATG, Post-Transplant Cyclophosphamide (PTCy), Abatacept and graft manipulation. Despite the high rate, first-line treatment for aGVHD is represented by corticosteroids, and Ruxolitinib is the standard second-line therapy; investigational approaches include Microbiota transplant and the infusion of Mesenchymal stem cells. GVHD is a pleiotropic disease involving any anatomical district; also, Ruxolitinib represents the standard for steroid-refractory cGVHD in this setting. It is a pleiotropic disease involving any anatomical district; also, Ruxolitinib represents the standard for steroid-refractory cGVHD in this setting. Extracorporeal Photopheresis (ECP) is still an option used for steroid refractoriness or to achieve a steroid-sparing. For Ruxolitinib-refractory cGVHD, Belumosudil and Axatilimab represent the most promising agents. Bronchiolitis obliterans syndrome (BOS) still represents a challenge; among the compounds targeting non-immune effectors, Alvelestat, a Neutrophil elastase inhibitor, seems promising in BOS. Finally, in both aGVHD and cGVHD, the association of biological markers with specific disease manifestations could help refine risk stratification and the availability of reliable biomarkers for specific treatments. [ABSTRACT FROM AUTHOR]

Previously, elastase activities have been found in some microorganisms and presented by metalloproteases, which may play an important role in helping bacterium invasiveness and establishment of infection. The elastases from Bacillus species are serine proteases, however, to date there are few reports on systematic study of these enzymes producers among representatives of Bacillus, as well as studies of their properties. The aim of this work was to study some physicochemical characteristics of partially purified elastase enzyme from a number of Bacillus sp. strains. Methods. The objects of the study were strains of Bacillus sp. (L1, L2, L9) isolated from the dry grass of the coastal zone of the Kinburn split (Mykolaiv region). Cultures were grown under submerged cultivation at 28°C, with a shaking speed of the nutrient medium of 230 rpm for 2 days. Methods of determining elastase activity in the culture liquid supernatant were used. Results. It has been shown that Bacillus sp. L9, L1, and L2 are characterized by high levels of elastase activity (35.80, 28.0, and 33.80 U/mL, respectively). The maximum synthesis of elastase occurs on the second day of cultivation of the producer at 28 °C and shaking at 230 rpm. Maximum hydrolysis of elastin by Bacillus sp. L1 and L2 are carried out at 40 °C but at different pH optima. Thus, for the preparation of Bacillus sp. L1, optimal pH is 8, and for L2 - 10. For the complex enzyme preparation of Bacillus sp. L9, maximum hydrolysis of elastin occurs at pH 9 and temperature 50 °C. [ABSTRACT FROM AUTHOR]

Alpha-1-antitrypsin deficiency is a genetic disorder caused by mutations in the SERPINA1 gene encoding alpha-1-antitrypsin (AAT), the major serine protease inhibitor in plasma. Reduced AAT levels are associated with elevated risk of developing emphysema mainly due to uncontrolled activity of neutrophil elastase in the lungs. The prevalent Z-AAT mutant and many rare pathogenic AAT variants also predispose to liver disease due to their accumulation as polymeric chains in hepatocytes. Part of these polymers are secreted into the bloodstream and could represent biomarkers of intra-hepatic accumulation. Moreover, being inactive, they further lower lung protection against proteases. Aim of our study is to accurately quantify the percentage of circulating polymers (CP) in a cohort of subjects with different SERPINA1 genotypes. CP concentration was measured in plasma or Dried Blood Spot (DBS) by a sensitive sandwich ELISA based on capture by the polymer-specific 2C1 monoclonal antibody. CP were significantly elevated in patients with the prevalent PI*SZ and PI*ZZ genotypes, with considerable intra-genotype variability. Notably, higher percentage of polymers was observed in association with elevated C-reactive protein. CP levels were also increased in carriers of the Mmalton variant, and of Mprocida, I, Plowell and Mherleen in heterozygosity with Z-AAT. These findings highlight the importance of implementing CP quantification in a clinical laboratory. Indeed, the variable amount of CP in patients with the same genotype may correlate with the variable severity of the associated lung and liver diseases. Moreover, CP can reveal the polymerogenic potential of newly discovered ultrarare AAT variants. [ABSTRACT FROM AUTHOR]

The research introduces a novel method for creating drug-loaded hydrogel beads that target anti-aging, anti-oxidative, and anti-inflammatory effects, addressing the interconnected processes underlying various pathological conditions. The study focuses on the development of hydrogel beads containing anti-aging compounds, antioxidants, and anti-inflammatory drugs to effectively mitigate various processes. The synthesis, characterization and in vitro evaluations, and potential applications of these multifunctional hydrogel beads are discussed. A polymeric alginate-orange peel extract (1:1) hydrogel was synthesized for encapsulating fish oil. Beads prepared with variable fish oil concentrations (0.1, 0.3, and 0.5 ml) were characterized, showing no significant decrease in size i.e., 0.5 mm and a reduction in pore size from 23 to 12 µm. Encapsulation efficiency reached up to 98% within 2 min, with controlled release achieved upto 45 to 120 min with increasing oil concentration, indicating potential for sustained delivery. Fourier-transform infrared spectroscopy confirmed successful encapsulation by revealing peak shifting, interaction between constituents. In vitro degradation studies showed the hydrogel's biodegradability improved from 30 to 120 min, alongside anti-inflammatory, anti-oxidative, anti-collagenase and anti-elastase activities, cell proliferation rate enhanced after entrapping fish oil. In conclusion, the synthesized hydrogel beads are a promising drug delivery vehicle because they provide stable and effective oil encapsulation with controlled release for notable anti-aging and regenerative potential. Targeted delivery for inflammatory and oxidative stress-related illnesses is one set of potential uses. Further research may optimize this system for broader applications in drug delivery and tissue engineering. [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]

This scientific article explores the potential use of C-terminal alpha-1-antitrypsin peptides (CAAPs) as biomarkers for predicting mortality in critically ill COVID-19 patients. The study found that CAAPs showed a higher predictive value for mortality compared to other commonly used parameters. The authors suggest that more research is needed to validate these findings and understand the biological mechanisms behind severe disease courses. The study was conducted ethically and with informed consent from participants. [Extracted from the article]

Neutrophil infiltration occurs in a variety of liver diseases, but it is unclear how neutrophils and hepatocytes interact. Neutrophils generally use granule proteases to digest phagocytosed bacteria and foreign substances or neutralize them in neutrophil extracellular traps. In certain pathological states, granule proteases play a destructive role against the host as well. More recently, nondestructive actions of neutrophil granule proteins have been reported, such as modulation of tissue remodeling and metabolism. Here, we report a completely different mechanism by which neutrophils act nondestructively, by inserting granules directly into hepatocytes. Specifically, elastase-containing granules were transferred to hepatocytes where elastase selectively degraded intracellular calcium channels to reduce cell proliferation without cytotoxicity. In response, hepatocytes increased expression of Serpin E2 and A3, which inhibited elastase activity. Elastase insertion was seen in patient specimens of alcohol-associated hepatitis, and the relationship between elastase-mediated ITPR2 degradation and reduced cell proliferation was confirmed in mouse models. Moreover, neutrophils from patients with alcohol-associated hepatitis were more prone to degranulation and more potent in reducing calcium channel expression than neutrophils from healthy individuals. This nondestructive and reversible action on hepatocytes defines a previously unrecognized role for neutrophils in the transient regulation of epithelial calcium signaling mechanisms. [ABSTRACT FROM AUTHOR]

Human neutrophil elastase (HNE) plays a key role in initiating inflammation in the cardiopulmonary and systemic contexts. Pathological auto-proteolysed two-chain (tc) HNE exhibits reduced binding affinity with inhibitors. Using AutoDock Vina v1.2.0, 66 flavonoid inhibitors, sivelestat and alvelestat were docked with single-chain (sc) HNE and tcHNE. Schrodinger PHASE v13.4.132 was used to generate a 3D-QSAR model. Molecular dynamics (MD) simulations were conducted with AMBER v18. The 3D-QSAR model for flavonoids with scHNE showed r2 = 0.95 and q2 = 0.91. High-activity compounds had hydrophobic A/A2 and C/C2 rings in the S1 subsite, with hydrogen bond donors at C5 and C7 positions of the A/A2 ring, and the C4' position of the B/B1 ring. All flavonoids except robustaflavone occupied the S1'-S2' subsites of tcHNE with decreased AutoDock binding affinities. During MD simulations, robustaflavone remained highly stable with both HNE forms. Principal Component Analysis suggested that robustaflavone binding induced structural stability in both HNE forms. Cluster analysis and free energy landscape plots showed that robustaflavone remained within the sc and tcHNE binding site throughout the 100 ns MD simulation. The robustaflavone scaffold likely inhibits both tcHNE and scHNE. It is potentially superior to sivelestat and alvelestat and can aid in developing therapeutics targeting both forms of HNE. [ABSTRACT FROM AUTHOR]

Digestive midgut protease activity in fifth instar larvae (L5) of Helicoverpa armigera was investigated under controlled conditions, where, an artificial diet containing different concentrations (0.005 µg/g - 0.3 µg/g) of Cry1Ac (Crystalline ä-endotoxin from class A, subclass c) toxin was fed. Here, the Bacillus thuringiensis (Bt) cotton seeds were used as Cry1Ac toxin source that was determined using Envirologix ELISA (Enzyme Linked Immunosorbent Assay) kit. The results showed an increase in the general midgut proteases as well as specific proteases activity linked to enzymes such as trypsin-, chymotrypsin-, elastase-, and leucine aminopeptidase-like with the increasing toxin content of the diet. However, carboxypeptidase-A and carboxypeptidase-B protease activity remained unaltered as a consequence of the incorporation of Cry1Ac toxin in the diet, which is also reported to be absent in midgut of H. armigera as evident in the control group. [ABSTRACT FROM AUTHOR]

Casein (CN) is the major family of proteins in milk and can cause allergic reactions in some infants and children. This study investigated the regulatory mechanisms of three animal‐derived proteases on allergenic epitopes. The results showed that all three proteases effectively reduced the antigenicity of CN. After hydrolysis, the secondary structure of CN underwent a conformational transition from α‐helix to β‐turn. Based on peptidomics analysis, the allergenic epitopes of αs1‐CN at positions 91–110, 131–151, 166–180 and 188–201 were effectively cleaved by pepsin, and the allergenic epitopes of β‐CN at positions 60–69, 139–158 and 182–194 were effectively cleaved by pancreatic elastase. [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]

Neutrophil elastase (HNE), like other members of the so-called GASPIDs (Granule-Associated Serine Peptidases of Immune Defense), is activated during protein biosynthesis in myeloid precursors and stored enzymatically active in cytoplasmic granules of resting neutrophils until secreted at sites of host defense and inflammation. Inhibitors thus could bind to the fully formed active site of the protease intracellularly in immature progenitors, in circulating neutrophils, or to HNE secreted into the extracellular space. Here, we have compared the ability of a panel of diverse inhibitors to inhibit HNE in the U937 progenitor cell line, in human blood-derived neutrophils, and in solution. Most synthetic inhibitors and, surprisingly, even a small naturally occurring proteinaceous inhibitor inhibit HNE intracellularly, but the extent and dynamics differ markedly from classical enzyme kinetics describing extracellular inhibition. Intracellular inhibition of HNE potentially affects neutrophil functions and has side effects, but it avoids competition of inhibitors with extracellular substrates that limit its efficacy. As both intra- and extracellular inhibition have advantages and disadvantages, the quantification of intracellular inhibition, in addition to classical enzyme kinetics, will aid the design of novel, clinically applicable HNE inhibitors with targeted sites of action. [ABSTRACT FROM AUTHOR]

Recently developed small-molecule inhibitors of the lysosomal protease dipeptidyl peptidase 1 (DPP1), also known as cathepsin C (CatC), can suppress suppurative inflammation in vivo by blocking the processing of zymogenic (pro-) forms of neutrophil serine proteases (NSPs), including neutrophil elastase, proteinase 3, and cathepsin G. DPP1 also plays an important role in activating granzyme serine proteases that are expressed by cytotoxic T lymphocytes (CTL) and natural killer (NK) cells. Therefore, it is critical to determine whether DPP1 inhibition can also cause off-target suppression of CTL/NK-cellmediated killing of virus-infected or malignant cells. Herein, we demonstrate that the processing of human granzymes A and B, transitioning from zymogen to active proteases, is not solely dependent on DPP1. Thus, the killing of target cells by primary human CD8+ T cells, NK cells, and gene-engineered anti-CD19 CAR T cells was not blocked in vitro even after prior exposure to high concentrations of the reversible DPP1 inhibitor brensocatib. Consistent with this observation, the turnover of model granzyme A/B peptide substrates in the human CTL/NK cell lysates was not significantly reduced by brensocatib. In contrast, preincubation with brensocatib almost entirely abolished (>90%) both the cytotoxic activity of mouse CD8+ T cells and granzyme substrate turnover. Overall, our finding that the effects of DPP1 inhibition on human cytotoxic lymphocytes are attenuated in comparison to those of mice indicates that granzyme processing/activation pathways differ between mice and humans. Moreover, the in vitro data suggest that human subjects treated with reversible DPP1 inhibitors, such as brensocatib, are unlikely to experience any appreciable deficits in CTL/NK-cellmediated immunities. [ABSTRACT FROM AUTHOR]

In diabetic ulcers, an increased secretion of human neutrophil elastase (HNE) and bacterial infections play crucial roles in hindering healing. Considering that, the present study proposed the development of multi-action polycaprolactone (PCL)/polyethylene glycol (PEG) electrospun fibers incorporating elastase-targeting peptides, AAPV and WAAPV, via blending. Characterization confirmed WAAPV's efficacy in regulating proteolytic enzymes by inhibiting HNE. The engineered fibers, particularly those containing PEG, exhibited optimal wettability but an accelerated degradation that was mitigated with the peptide's inclusion, thus promoting a sustained peptide release over 24 h. Peptide loading was verified indirectly through thermal stability and hydration capacity studies (hydrophobic bonding between PCL and WAAPV and hydrophilic affinities between PCL/PEG and AAPV) and determined at ≈51.1 µg/cm2 and ≈46.0 µg/cm2 for AAPV and ≈48.5 µg/cm2 and ≈51.3 µg/cm2 for WAAPV, respectively, for PCL and PCL/PEG. Both AAPV and WAAPV effectively inhibited HNE, with PEG potentially enhancing this effect by interacting with the peptides and generating detectable peptide–PEG complexes (≈10% inhibition with PCL + peptide fibers after 6 h of incubation, and ≈20% with PCL/PEG + peptide fibers after 4 h incubation). Peptide-loaded fibers demonstrated antibacterial efficacy against Staphylococcus aureus (up to ≈78% inhibition) and Escherichia coli (up to ≈66% inhibition), with peak effectiveness observed after 4 and 2 h of incubation, respectively. This study provides initial insights into the WAAPV's potential for inhibiting HNE and bacteria activities, showing promise for applications in diabetic ulcer management. [ABSTRACT FROM AUTHOR]

In our study, using chromatographic techniques, we isolated three bioactive compounds, which were structurally elucidated as (E)-2-(3-(3,4-dimethoxyphenyl)acrylamido)-N-methylbenzamide (1), 4-Hydroxyquinoline-2-carboxylic acid (2), and (E)-2-Cyano-3-(4-hydroxyphenyl)acrylic acid (3), using spectroscopic methods. The anti-melanogenic, anti-inflammatory, antioxidant, and anti-aging properties were evaluated in vitro by measuring the activity of pharmacological targets including tyrosinase, melanin, NF-κB, hyaluronidase, elastase, collagenase, and Nrf2. Our results show that compound 1 is the most active with IC50 values of 14.19 μM (tyrosinase inhibition), 22.24 μM (melanin inhibition), 9.82–12.72 μM (NF-κB inhibition), 79.71 μM (hyaluronidase inhibition), 80.13 μM (elastase inhibition), 76.59 μM (collagenase inhibition), and 116–385 nM (Nrf2 activation) in the THP-1, HEK001, WS1, and HMCB cells. These findings underscore the promising profiles of the aqueous extract of R. urticifolius at safe cytotoxic concentrations. Additionally, we report, for the first time, the isolation and characterisation of these nitrogenous compounds in the R. urticifolius species. Finally, compound 1, isolated from R. urticifolius, is a promising candidate for the development of more effective and safer compounds for diseases related to skin pigmentation, protection against inflammation, and oxidative stress. [ABSTRACT FROM AUTHOR]

Background: Cystic fibrosis (CF) is an autosomal recessive disorder affecting the exocrine functions of many organs including the pancreas. Clinical and laboratory recognition of pancreatic insufficiency is important for proper treatment. Early radiologic detection of pancreatic pathologic changes can help better evaluation. Objectives: To correlate exocrine pancreatic function with pancreatic ultrasound changes in CF children. Methods: This was a prospective cross-sectional study to assess the exocrine pancreatic function by measuring fecal elastase-1 (FE-1) level and doing pancreatic ultrasound for CF patients. Results: A total of 30 CF patients with age range from 3 months to 10 years were included in the study. They were categorized into three age groups; below one year, from 1 to 4 years and above 4 years old. None of our cases was exocrine pancreatic sufficient regarding FE-1 level and most of them (86.7%) had severe fecal elastase deficiency. Before 1 year of age there was no detected abnormal ultrasound findings in pancreas; only 8 older patients had abnormal US detected as hyper-echogenicity of pancreas. Abnormal US was more detected among patients with severe pancreatic insufficiency. The sensitivity of ability of fecal elastase level to predict abnormal ultrasound was 80%, with 84% specificity at cut of point 15.2 µg/g. Conclusions: Fecal-elastase-1 measurement in cystic fibrosis children can diagnose the exocrine pancreatic insufficiency even without clinical manifestations of steatorrhea. FE-1 level can predict pathologic changes of pancreas detected by transabdominal ultrasound. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the effects of elicitors on Vanda coerulea Griff. Ex Lindl. protocorms to enhance bioactive compound production and evaluate their biological activities relevant to cosmeceutical applications. The protocorms were developed from the callus treated with different elicitors, including 6-benzylaminopurine (BA), methyl jasmonate (MeJA), and chitosan. Both the adult plant and protocorms were extracted by maceration in 80% methanol and investigated for their chemical compositions using high-performance liquid chromatography. The extracts were evaluated for antioxidant, anti-collagenase, anti-elastase, and anti-tyrosinase activities. In addition, anti-inflammatory properties were assessed using a real-time polymerase chain reaction. The irritation potency was evaluated using the hen's egg test-chorioallantoic membrane test. The findings revealed that protocorms treated with BA and chitosan developed a greener color, while those treated with MeJA exhibited a distinct darker coloration. Elicitation with BA and chitosan resulted in protocorms with comparable or higher levels of syringic acid, rutin, and quercin compared with the adult plant, with rutin being the most prominent identified compound. Furthermore, rutin was reported as the compound responsible for all biological activities. The chitosan-treated protocorm extract exhibited potent inhibition against oxidation, collagenase, elastase, tyrosinase, and inflammatory cytokines, along with a nonirritating effect, making it a promising candidate for cosmeceutical applications. [ABSTRACT FROM AUTHOR]

This document provides summaries of several patent applications and research studies related to drug repurposing for various medical conditions. The patent applications discuss the potential use of different drugs for the treatment of psoriasis, nonalcoholic steatohepatitis, pain, frozen shoulder syndrome, anxiety disorders, addiction to stimulant drugs, and inherited epidermolysis bullosa. The research studies focus on using epidermal growth factor for interstitial cystitis, fenoldopam for complications of autosomal dominant polycystic kidney disease, aldose reductase inhibitors for muscle regeneration, and a combination of aldose reductase inhibitors and nucleosides for phosphomannomutase 2 deficiency. These findings offer potential treatment options, but further research is needed. [Extracted from the article]

Neutrophil proteinase 3 (PR3) is an important drug target for inflammatory lung diseases such as chronic obstructive pulmonary disease and cystic fibrosis. Drug discovery efforts targeting PR3 require active enzyme for in vitro characterization, such as inhibitor screening, enzymatic assays, and structural studies. Recombinant expression of active PR3 overcomes the need for enzyme supplies from human blood and in addition allows studies on the influence of mutations on enzyme activity and ligand binding. Here, we report the expression of recombinant PR3 (rPR3) using a baculovirus expression system. The purification and activation process described resulted in highly pure and active PR3. The activity of rPR3 in the presence of commercially available inhibitors was compared with human PR3 by using a fluorescence-based enzymatic assay. Purified rPR3 had comparable activity to the native human enzyme, thus being a suitable alternative for enzymatic studies in vitro. Further, we established a surface plasmon resonance-based assay to determine binding affinities and kinetics of PR3 ligands. These methods provide valuable tools for early drug discovery aiming towards treatment of lung inflammation. [ABSTRACT FROM AUTHOR]

Background: Numerous immunoassays have been commercialized to determine pancreatic elastase (PE) in feces in screening for exocrine pancreatic insufficiency (EPI), but how the different assays compare to one another is controversial, especially in the context that all methods use the same cut-off values for interpreting the results obtained on the presence or absence of EPI or the degree of insufficiency if it is present. Our aim was to analytically verify a new method for determining PE, compare the results with a previous method, and verify the declared cut-off values for interpretation of the results. Methods: PE in the stool was assayed using a previous monoclonal enzyme-linked immunosorbent assay ("ScheBo ELISA") and a new polyclonal particle-enhanced turbidimetric immunoassay ("Bühlmann PETIA"). The direct method comparison of two immunoassays was performed in 40 samples. Clinical comparisons were conducted against each other for the binary determination of "abnormal/normal" elastase levels and the three-way determination of "severe/moderate/no" EPI in 56 samples. The indirect comparison method used external quality assessment (EQA) data to compare the monoclonal and polyclonal immunoassays for PE, and additionally compare the monoclonal ScheBo ELISA to a monoclonal chemiluminescence immunoassay ("DiaSorin CLIA"). Results: Precision in the series and intra-laboratory precision for Bühlmann PETIA met the manufacturer's specifications for the concentration range of limit/lower values and the range of normal values. The Bühlmann PETIA immunoassay on different analytical platforms yielded comparable results and nearly perfect agreement in the case of three-way classification (kappa = 0.89 with 95%CI from 0.79 to 1.00. ScheBo ELISA tends to generate higher values of pancreatic elastase than the Bühlmann PETIA; agreement between the methods was moderate in the case of binary classification (kappa = 0.43; 95% CI 0.25 to 0.62), and substantial in the case of three-way classification (kappa = 0.62; 95% CI 0.50 to 0.75). EQA data analysis showed a statistically significant difference between ScheBo ELISA and Bühlmann PETIA peer groups (p = 0.031), as well as the DiaSorin CLIA and ScheBo ELISA peer groups (p = 0.010). Conclusion: The ScheBo ELISA and Bühlmann PETIA do not appear to be commutable in the analytical and clinical context. Our data address a discordance between different mono- and polyclonal immunoassays for pancreatic elastase and the potential of misclassification using its universal cut-off values in screening suspected patients for exocrine pancreatic insufficiency. [ABSTRACT FROM AUTHOR]

Balaustium murorum (Acariformes: Erythraeidae) is a pollen feeder and predatory mite that occasionally causes dermatitis. We sought to determine the protease profile in the whole-mite extract of B. murorum using general and specific substrates as well as inhibitors. The pH profiles using two general substrates, azacasein and haemoglobin, revealed an optimal pH of 6 for both substrates. Specific trypsin inhibitor TLCK showed no influence activity towards BApNA; however, TPCK and AEBSF inhibited chymotrypsin and elastase, respectively. Specific cysteine protease inhibitors Cystatin and E-64 influenced both types of cathepsins B and L, while DTT as a specific activator increased the enzymatic activities. Only aminopeptidase showed significant activity and inhibition using negative control and phenanthroline. Chymotrypsin and elastase had the highest activity at pH of 9–10 and 10, respectively, by using both substrates alone and substrates along with specific inhibitors. Cathepsins B and L had pH optima of 6. Aminopeptidase revealed a broad pH optimum of 5–7 on both substrates HA and HPA alone and substrate along with inhibitor phenanthroline. Finally, different concentrations of each inhibitor were used to find IC50 values of TPCK, AEBSF, cystatin, E-64, DTT and phenanthroline required to inhibit half of enzyme activity. [ABSTRACT FROM AUTHOR]

Pulmonary emphysema is a primary component of chronic obstructive pulmonary disease (COPD), a life-threatening disorder characterized by lung inflammation and restricted airflow, primarily resulting from the destruction of small airways and alveolar walls. Cumulative evidence suggests that nicotinic receptors, especially the α7 subtype (α7nAChR), is required for anti-inflammatory cholinergic responses. We postulated that the stimulation of α7nAChR could offer therapeutic benefits in the context of pulmonary emphysema. To investigate this, we assessed the potential protective effects of PNU-282987, a selective α7nAChR agonist, using an experimental emphysema model. Male mice (C57BL/6) were submitted to a nasal instillation of porcine pancreatic elastase (PPE) (50 µl, 0.667 IU) to induce emphysema. Treatment with PNU-282987 (2.0 mg/kg, ip) was performed pre and post-emphysema induction by measuring anti-inflammatory effects (inflammatory cells, cytokines) as well as anti-remodeling and anti-oxidant effects. Elastase-induced emphysema led to an increase in the number of α7nAChR-positive cells in the lungs. Notably, both groups treated with PNU-282987 (prior to and following emphysema induction) exhibited a significant decrease in the number of α7nAChR-positive cells. Furthermore, both groups treated with PNU-282987 demonstrated decreased levels of macrophages, IL-6, IL-1β, collagen, and elastic fiber deposition. Additionally, both groups exhibited reduced STAT3 phosphorylation and lower levels of SOCS3. Of particular note, in the post-treated group, PNU-282987 successfully attenuated alveolar enlargement, decreased IL-17 and TNF-α levels, and reduced the recruitment of polymorphonuclear cells to the lung parenchyma. Significantly, it is worth noting that MLA, an antagonist of α7nAChR, counteracted the protective effects of PNU-282987 in relation to certain crucial inflammatory parameters. In summary, these findings unequivocally demonstrate the protective abilities of α7nAChR against elastase-induced emphysema, strongly supporting α7nAChR as a pivotal therapeutic target for ameliorating pulmonary emphysema. [ABSTRACT FROM AUTHOR]

Background: P. aeruginosa, a significant bacterium, can cause severe illness and resistance to antibiotics. Quorum sensing (QS) systems regulate virulence factors production. Targeting QS could reduce bacteria pathogenicity and prevent antibiotic resistance. Cruciferous vegetables contain sulforaphane, known for its anti-inflammatory, antioxidant, anticancer, and antimicrobial properties. Aim: We aimed to examine the inhibitory influences of sulforaphane, at a subinhibitory concentration (¼ minimum inhibitory concentration, MIC), on virulence and QS in P. aeruginosa. Materials and methods: The sulforaphane’s anti-virulence actions at subinhibitory concentrations were explored in vitro and in vivo. A sub-MIC concentration of sulforaphane was combined with anti-pseudomonal drugs, and the results of this combination were assessed. The virtual affinity of sulforaphane for the receptors of QS was studied, and its effect on the expression of QS genes was quantified. Results: Sulforaphane significantly decreased the biofilm formation, motility, ability to withstand oxidative stress, and the synthesis of virulence extracellular enzymes such as proteases, hemolysins, and elastase, as well as other virulence factors like pyocyanin. In addition, sulforaphane lessened the severity of P. aeruginosa infection in mice. Sulforaphane reduced the antipseudomonal antibiotics’ MICs when used together, resulting in synergistic effects. The observed anti-virulence impacts were attributed to the ability of sulforaphane to inhibit QS via suppressing the QS genes’ expression. Conclusion: Sulforaphane shows promise as a potent anti-virulence and anti-QS agent that can be used alongside conventional antimicrobials to manage severe infections effectively. Furthermore, this study paves the way for further investigation of sulforaphane and similar structures as pharmacophores for antiQS candidates. [ABSTRACT FROM AUTHOR]

According to the "Evaluation Specification for Cosmetic Efficacy Claims", the results of laboratory tests can be used to support multiple types of cosmetic efficacy claims, including anti-wrinkle, firming, soothing, oil control, exfoliating, anti-hair breakage, dandruff removal, moisturizing, hair care, raw material efficacy, and declared mild (non irritating), and quantitative indicators. The laboratory tests are different from the actual usage status of the product, and their appeal to consumers are far less than that of human research. They also have the advantages of good repeatability, simple, low cost, et al. Laboratory tests include physicochemical (biochemical) tests, cell tests, skin model tests and animal tests, et al. Although it may be difficult to accurately extrapolate the effects of physicochemical methods to the actual use of the human body, they are widely used now. Through testing the inhibitory effect of cosmetics in matrix metalloproteinase, elastase and nonenzymatic glycosylation reaction, or the scavenging ability of cosmetics in free radicals, the anti-wrinkle efficacy of the product can be evaluated. Through testing the inhibitory effect of cosmetics in hyaluronidase, or the scavenging ability of cosmetics in free radicasl, the soothing efficacy of the product can be evaluated. Through testing the moisture absorption and moisturizing rate of cosmetics, the soothing and moisturizing efficacy of the product can be evaluated. Some research also show that there is a certain positive correlation between the moisturizing rate of physicochemical testing methods and human research. Through testing the ability of waterproof and anti-sweat, the hold on efficacy of makeup can be evaluated. At present, there are various physicochemical methods for evaluating the efficacy of cosmetics widely used, but many methods lack standard specifications and guidance. The existing group standards have certain shortcomings or limitations, leading to some problems in practical application. For example, the claims of some efficacies lack appropriate physicochemical testing methods, the same test indicator corresponds to multiple efficacy claims, some standards are not applicable to certain products and need to be improved in sample pre-treatment and result analysis, and the test results can not support the efficacy claims scientifically. In the future, it is recommended to strengthen the standardization of physicochemical test methods, improve the standardized system for evaluating the efficacy of cosmetics, and to promote the development of the cosmetics industry. [ABSTRACT FROM AUTHOR]

Serine protease inhibitors are a superfamily of proteins that regulate various physiological processes including fibrinolysis, inflammation and immune responses. In parasite systems, serpins are believed to play important roles in parasite colonization, inhibition of host immune serine proteases and penetration of defensive barriers. However, serpins are less well characterized in schistosomes. In this study, a Schistosoma mansoni serpin (Smserpin-p46) containing a 1360 base pair open reading frame, was cloned, expressed and functionally characterized. Bioinformatics analysis revealed that Smserpin-p46 contains the key residues, structural domains and motifs characteristic of inhibitory serpins. Gene expression profiling demonstrated stage-specific expression of Smserpin-p46 with the highest expression in adult male worms. Recombinant Smserpin-p46 (rSmserpin-p46) inhibited both human neutrophil cathepsin G and elastase, key serine proteases involved in NETosis, a program for the formation of neutrophil extracellular traps. Using specific rabbit antiserum, Smserpin-p46 was detected in soluble worm antigen preparation and was localized to the adult worm tegument. Cumulatively, the expression of Smserpin-p46 on the parasite tegument and its ability to inhibit proteases involved in NETosis highlights the importance of this serpin in parasite-host interactions and encourages its further investigation as a candidate vaccine antigen for the control of schistosomiasis. [ABSTRACT FROM AUTHOR]

Proteins delivered by endocytosis or autophagy to lysosomes are degraded by exo- and endoproteases. In humans 15 lysosomal cathepsins (CTS) act as important physiological regulators. The cysteine proteases CTSB and CTSL and the aspartic protease CTSD are the most abundant and functional important lysosomal proteinases. Whereas their general functions in proteolysis in the lysosome, their individual substrate, cleavage specificity, and their possible sequential action on substrate proteins have been previously studied, their functional redundancy is still poorly understood. To address a possible common role of highly expressed and functional important CTS proteases, we generated CTSB-, CTSD-, CTSL-, and CTSBDL-triple deficient (KO) human neuroblastoma-derived SH-SY5Y cells and CTSB-, CTSD-, CTSL-, CTSZ and CTSBDLZ-quadruple deficient (KO) HeLa cells. These cells with a combined cathepsin deficiency exhibited enlarged lysosomes and accumulated lipofuscin-like storage material. The lack of the three (SH-SY5Y) or four (HeLa) major CTSs caused an impaired autophagic flux and reduced degradation of endocytosed albumin. Proteome analyses of parental and CTS-depleted cells revealed an enrichment of cleaved peptides, lysosome/autophagy-associated proteins, and potentially endocytosed membrane proteins like the amyloid precursor protein (APP), which can be subject to endocytic degradation. Amino- and carboxyterminal APP fragments accumulated in the multiple CTS-deficient cells, suggesting that multiple CTS-mediated cleavage events regularly process APP. In summary, our analyses support the idea that different lysosomal cathepsins act in concert, have at least partially and functionally redundant substrates, regulate protein degradation in autophagy, and control cellular proteostasis, as exemplified by their involvement in the degradation of APP fragments. [ABSTRACT FROM AUTHOR]

Aiming to study the performance, carcass characteristics, nutrient digestibility, blood parameters, salivary cortisol levels, and economic viability of pigs administered aspartic protease, a total of 135 pigs were housed in pens in a randomized block design, divided into five treatments with nine replications. The experimental diets were positive control (PC), basic diet with a 5.0% reduction in protein and amino acid requirements; negative control (NC) with a 7.5% reduction in protein and amino acid requirements; NC + 100 g/mT of aspartic protease (NC100); NC + 150 g/mT of aspartic protease (NC150); and NC + 200 g/mT of neutral serine protease (NC200). The inclusion of protease, independently of the source and amount, increased the average daily weight gain (P < 0.05) of animals compared with the control treatments (PC and NC), improved feed conversion (P < 0.05) in early stages, and improved diet digestibility (P < 0.05) compared with the PC. Treatment with NC150 and NC200 resulted in greater carcass weights (P < 0.05) than treatment with the PC. NC100 led to a greater carcass yield than PC (P < 0.05), and NC150 resulted in a greater loin eye area than PC (P < 0.05). No differences (P > 0.05) in the blood parameters or salivary cortisol levels were found. Regarding economic viability, proteases increased the profitability, with NC150 leading to the best results. Thus, the use of aspartic proteases is recommended to improve performance and further facilitate pork production. [ABSTRACT FROM AUTHOR]

Lipopolysaccharide-induced (LPS) inflammation is used as model to understand the role of inflammation in brain diseases. However, no studies have assessed the ability of peripheral low-level chronic LPS to induce neutrophil activation in the periphery and brain. Subclinical levels of LPS were injected intraperitoneally into mice to investigate its impacts on neutrophil frequency and activation. Neutrophil activation, as measured by CD11b expression, was higher in LPS-injected mice compared to saline-injected mice after 4 weeks but not 8 weeks of injections. Neutrophil frequency and activation increased in the periphery 4–12 h and 4–8 h after the fourth and final injection, respectively. Increased levels of G-CSF, TNFa, IL-6, and CXCL2 were observed in the plasma along with increased neutrophil elastase, a marker of neutrophil extracellular traps, peaking 4 h following the final injection. Neutrophil activation was increased in the brain of LPS-injected mice when compared to saline-injected mice 4–8 h after the final injection. These results indicate that subclinical levels of peripheral LPS induces neutrophil activation in the periphery and brain. This model of chronic low-level systemic inflammation could be used to understand how neutrophils may act as mediators of the periphery–brain axis of inflammation with age and/or in mouse models of neurodegenerative or neuroinflammatory disease. [ABSTRACT FROM AUTHOR]

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]

Pseudomonas aeruginosa isolates were recovered from surface river water samples in La Rioja region (Spain) to characterise their antibiotic resistance, molecular typing and virulence mechanisms. Fifty-two P. aeruginosa isolates were isolated from 15 different water samples (45.4%) and belonged to 23 different pulsed-field electrophoresis (PFGE) patterns. All isolates were susceptible to all antibiotics tested, except one carbapenem-resistant P. aeruginosa that showed a premature stop codon in OprD porin. Twenty-two sequence types (STs) (six new ones) were detected among 29 selected P. aeruginosa (one strain with a different PFGE pattern per sample), with ST274 (14%) being the most frequent one. O:6 and O:3 were the predominant serotypes (31%). Seven virulotypes were detected, being 59% exoS-exoY-exoT-exoA-lasA-lasB-lasI-lasR-rhlAB - rhlI - rhlR - aprA -positive P. aeruginosa. It is noteworthy that the exlA gene was identified in three strains (10.3%), and the exoU gene in seven (24.1%), exoS in 18 (62.1%), and both exoS and exoU genes in one strain. High motility ranges were found in these strains. Twenty-seven per cent of strains produced more biofilm biomass, 90% more pyorubin, 83% more pyocyanin and 65.5% more than twice the elastase activity compared with the PAO1 strain. These results highlight the importance of rivers as temporary reservoirs and sources of P. aeruginosa transmission, and show the importance of their epidemiological surveillance in the environment. [ABSTRACT FROM AUTHOR]

To investigate the effects of neutrophil elastase inhibitor (sivelestat sodium) on gastrointestinal function in sepsis. A reanalysis of the data from previous clinical trials conducted at our center was performed. Septic patients were divided into either the sivelestat group or the non‐sivelestat group. The gastrointestinal dysfunction score (GIDS), feeding intolerance (FI) incidence, serum levels of intestinal barrier function and inflammatory biomarkers were recorded. The clinical severity and outcome variables were also documented. A total of 163 septic patients were included. The proportion of patients with GIDS ≥2 in the sivelestat group was reduced relative to that in the non‐sivelestat group (9.6% vs. 22.5%, p = 0.047) on the 7th day of intensive care unit (ICU) admission. The FI incidence was also remarkably reduced in the sivelestat group in contrast to that in the non‐sivelestat group (21.2% vs. 37.8%, p = 0.034). Furthermore, the sivelestat group had fewer days of FI [4 (3, 4) vs. 5 (4–6), p = 0.008]. The serum levels of d‐lactate (p = 0.033), intestinal fatty acid‐binding protein (p = 0.005), interleukin‐6 (p = 0.001), white blood cells (p = 0.007), C‐reactive protein (p = 0.001), and procalcitonin (p < 0.001) of the sivelestat group were lower than those of the non‐sivelestat group. The sivelestat group also demonstrated longer ICU‐free days [18 (0–22) vs. 13 (0–17), p = 0.004] and ventilator‐free days [22 (1–24) vs. 16 (1–19), p = 0.002] compared with the non‐sivelestat group. In conclusion, sivelestat sodium administration appears to improve gastrointestinal dysfunction, mitigate dysregulated inflammation, and reduce disease severity in septic patients. [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]

Pseudomonas aeruginosa is an opportunistic pathogen that usually causes chronic infections and even death in patients. The treatment of P. aeruginosa infection has become more challenging due to the prevalence of antibiotic resistance and the slow pace of new antibiotic development. Therefore, it is essential to explore non-antibiotic methods. A new strategy involves screening for drugs that target the quorum-sensing (QS) system. The QS system regulates the infection and drug resistance in P. aeruginosa. In this study, veratryl alcohol (VA) was found as an effective QS inhibitor (QSI). It effectively suppressed the expression of QS-related genes and the subsequent production of virulence factors under the control of QS including elastase, protease, pyocyanin and rhamnolipid at sub-inhibitory concentrations. In addition, motility activity and biofilm formation, which were correlated with the infection of P. aeruginosa, were also suppressed by VA. In vivo experiments demonstrated that VA could weaken the pathogenicity of P. aeruginosa in Chinese cabbage, Drosophila melanogaster, and Caenorhabditis elegans infection models. Molecular docking, combined with QS quintuple mutant infection analysis, identified that the mechanism of VA could target the LasR protein of the las system mainly. Moreover, VA increased the susceptibility of P. aeruginosa to conventional antibiotics of tobramycin, kanamycin and gentamicin. The results firstly demonstrate that VA is a promising QSI to treat infections caused by P. aeruginosa. [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]

Neutrophil elastase (NE) is taken up by macrophages, retains intracellular protease activity, and induces a pro-inflammatory phenotype. However, the mechanism of NE-induced pro-inflammatory polarization of macrophages is not well understood. We hypothesized that intracellular NE degrades histone deacetylases (HDAC) and Sirtuins, disrupting the balance of lysine acetylation and deacetylation and resulting in nuclear to cytoplasmic translocation of a major alarmin, High Mobility Group Box 1 (HMGB1), a pro-inflammatory response in macrophages. Human blood monocytes were obtained from healthy donors or from subjects with cystic fibrosis (CF) or chronic obstructive pulmonary disease (COPD). Monocytes were differentiated into blood monocyte derived macrophages (BMDMs) in vitro. Human BMDMs were exposed to NE or control vehicle, and the abundance of HDACs and Sirtuins was determined by Western blotting of total cell lysates or nuclear extracts or determined by ELISA. HDAC, Sirtuin, and Histone acetyltransferase (HAT) activities were measured. NE degraded most HDACs and Sirtuin (Sirt)1, resulting in decreased HDAC and sirtuin activities, with minimal change in HAT activity. We then evaluated whether the NE-induced loss of Sirt activity or loss of HDAC activities would alter the cellular localization of HMGB1. NE treatment or treatment with Trichostatin A (TSA), a global HDAC inhibitor, both increased HMGB1 translocation from the nucleus to the cytoplasm, consistent with HMGB1 activation. NE significantly degraded Class I and II HDAC family members and Sirt 1, which shifted BMDMs to a pro-inflammatory phenotype. [ABSTRACT FROM AUTHOR]

• Seaweed (Hypnea pannosa) powder was incorporated to formulate the bathing bar. • A quality evaluation of the seaweed bathing bar was performed. • The bioactive properties, such as in-vitro cytotoxicity and the antibacterial, anti-inflammatory, anticancer, anti-wrinkling, and antiaging activity of the seaweed bathing bar, were analyzed. • The functional properties of the bathing bar were investigated. • Compared to the control bathing bar (CBB), the seaweed bathing bar (SBB) exhibited superior bioactive and physicochemical properties. The study aimed to formulate and evaluate the physicochemical characteristics, cytotoxicity assay, invitro anti-aging, anti-wrinkling, anti-cancer, anti-inflammatory, antibacterial activity, and functional properties of the Hypnea pannosa seaweed incorporated bathing bar. The study found that incorporating Hypnea pannosa seaweed powder improved the physicochemical characteristics of the bathing bar, with values within standard specifications. Seaweed showed no cytotoxic activity in the Vero cell line, with an IC 50 value of 435.67 µg/ml found safe for the bathing bar formulation. The seaweed bathing bar (SBB) had high values for Total fatty matter (40.54±0.02 %), Free Caustic Alkali (0.00 %), Moisture (7.97±0.01 %), pH (8.10), and Lather (230 ml). The SBB also exhibited potent cytotoxicity (12.83 %) against the B16F10 cell line and was significantly more effective than the control bathing bar (CBB) at inhibiting elastase with the IC 50 value of 76.10 µg/ml and collagenase (85.85 µg/ml) in the SKMEL cell line. The inhibition rate of egg albumin denaturation increased gradually with both SBB and CBB concentrations, with significantly higher inhibition in SBB (62.1 %) compared to CBB (17.5 %) at maximum concentration. Additionally, the SBB demonstrated superior antibacterial activity against Staphylococcus aureus, Pseudomonas aeruginosa, and E. coli but was least effective against Bacillus cereus. The SBB contains polyphenols that have a significant use in cosmeceuticals, as revealed by functional analysis (3569.7 - 3473.4 cm-1). The SBB has potential applications in treating skin infections and in anti-aging, anti-wrinkling, anti-inflammatory, and anticancer agents. SBB can potentially revolutionize the skincare and pharmaceutical industry by providing a natural and sustainable alternative to synthetic ingredients commonly used in these products. Its unique properties, including high levels of antioxidants and anti-inflammatory compounds, may also hold promise for treating various skin conditions and diseases. Incorporating seaweed into skincare and pharmaceutical products could benefit consumers and the environment. [ABSTRACT FROM AUTHOR]

This study aimed to determine the anti-collagenase, anti-elastase, and anti-hyaluronidase activities of Passiflora quadrangularis fruit extracts (epicarp, mesocarp, endocarp, and seed), develop stable nanoemulsions, and evaluate the efficacy of the nanoemulsions containing extracts in human volunteers. The results indicated that the epicarp and seed extracts exhibited collagenase, elastase, and hyaluronidase inhibition effects. Gallic acid was identified in the extracts, with the highest concentration found in the endocarp extract (1449.35 mg gallic acid/g extract) followed by the seed extract (839.63 mg gallic acid/g extract). The endocarp and seed extracts demonstrated good stability at different temperatures. Consequently, these extracts were selected for incorporation into nanoemulsions due to their high yield, excellent anti-tyrosinase and anti-aging activity, and good stability, making them suitable for cosmetic products. The nanoemulsions were prepared using the ultrasonication method, resulting in a particle size of 133.1 ± 0.8 nm, a polydispersity index of 0.33 ± 0.03, and a zeta potential of −61.8 ± 0.6 mV, indicating good stability. The nano-serum containing extracts was non-irritating and safe for the skin. Skin evaluation among human volunteers after 60 days of application revealed improvements in skin hydration and reduction in wrinkles. In conclusion, nanoemulsions containing P. quadrangularis fruit extracts have the potential to be effective cosmetic products. [ABSTRACT FROM AUTHOR]

This document is an editorial from the International Journal of Molecular Sciences titled "Lysosomal Proteases and Their Inhibitors." It provides an overview of the role and function of lysosomal proteases, specifically cathepsins, in various physiological and pathological processes. The editorial highlights the importance of regulating the potentially harmful activity of these enzymes outside the lysosomes through pH, zymogen activation, and protein inhibitors. The document also includes summaries of original articles and reviews that delve deeper into the field, discussing topics such as the evolution of the papain family, the involvement of cathepsins in viral infections, the therapeutic potential of cathepsin K, and the functions of Kunitz-type inhibitors in parasites and ticks. The editorial concludes by emphasizing the recent advances in understanding lysosomal proteases and their inhibitors and their significance as diagnostic and therapeutic targets for various diseases. [Extracted from the article]

The article focuses on examining the gene expression signature of Alpha-1 Antitrypsin Deficiency (AATD) across different lung cell types using single-cell RNA-sequencing (scRNA-seq) data. It highlights the association between AATD and proinflammatory changes in lung cells, especially in alveolar macrophages and other cell types.

Introduction: The development of bioconjugates for the targeted delivery of anticancer agents is gaining momentum after recent success of antibody drug conjugates (ADCs) in the clinic. Smaller format conjugates may have several advantages including better tumor penetration; however, cellular uptake and trafficking may be substantially different from ADCs. To fully leverage the potential of small molecule drug conjugates (SMDCs) with potent binding molecules mediating tumor homing, novel linker chemistries susceptible for efficient extracellular activation and payload release in the tumor microenvironment (TME) need to be explored. Methods: We designed a novel class of SMDCs, which target αvβ3 integrins for tumor homing and are cleaved by neutrophil elastase (NE), a serine protease active in the TME. A peptidomimetic αvβ3 ligand was attached via optimized linkers composed of substrate peptide sequences of NE connected to different functional groups of various payload classes, such as camptothecins, monomethyl auristatin E, kinesin spindle protein inhibitors (KSPi) and cyclindependent kinase 9 inhibitors (CDK-9i). Results: NE-mediated cleavage was found compatible with the diverse linker attachments via hindered ester bonds, amide bonds and sulfoximide bonds. Efficient and traceless release of the respective payloads was demonstrated in biochemical assays. The newly designed SMDCs were highly stable in buffer as well as in rat and human plasma. Cytotoxicity of the SMDCs in cancer cell lines was clearly dependent on NE. IC50 values were in the nanomolar or subnanomolar range across several cancer cell lines reaching similar potencies as compared to the respective payloads only in the presence of NE. In vivo pharmacokinetics evaluating SMDC and free payload exposures in rat and particularly the robust efficacy with good tolerability in triple negative breast and small cell lung cancer murine models demonstrate the utility of this approach for selective delivery of payloads to the tumor. Discussion: These results highlight the broad scope of potential payloads and suitable conjugation chemistries paving the way for future SMDCs harnessing the safety features of targeted delivery approaches in combination with NE cleavage in the TME. [ABSTRACT FROM AUTHOR]