Elastase Enzyme Research Articles

Variation of elastase, collagenase, tyrosinase enzyme inhibitory and antioxidant potential of different tea cultivars

Sri Lankan tea germplasm consists of over 600 accessions, and currently, 70 cultivars are recommended for commercial cultivation. However, the skin whitening and anti-aging effects of Sri Lankan tea cultivars/accessions have not been reported. In this study, tea leaves and black tea of fifteen tea cultivars were tested for their anti-oxidative and enzyme inhibitory potentials against elastase, collagenase, and tyrosinase enzymes for the first time. All samples were evaluated for their total polyphenol, caffeine, and individual catechin contents, and the results significantly differed among tea cultivars. The antioxidant potential of black tea and tea leaves significantly varied among the tea cultivars used. The enzyme inhibitory potential on tyrosinase, elastase, and collagenase inhibitory potential in black tea were varied among the tea cultivars, with the IC50 ranged from 208.82 ± 5.18 to ≥ 1200 ppm, 36.71 ± 6.38 to ≥ 600 ppm, and 786.31 ± 2.33 to ≥ 1300 ppm, respectively. None of the tested tea cultivars resulted in strong collagenase inhibitory potential. This study concludes that, among the tested tea cultivars, black tea manufactured using tea cultivars TRI 4004 and TRI 4049 and both tea leaves and black tea manufactured from TRI 4053, TRI 3017, and TRI 4061 exhibited remarkable anti-aging and skin-whitening properties. The findings of this study could be utilized for the selection of tea cultivars/accessions for the manufacture of speciality tea with functional properties and also to promote Sri Lankan tea in the global market.

Stachys byzantina K. Koch (Lamiaceae) as a potential ingredient for delaying skin ageing and treating hyperpigmentation disorders in pharmaceutical products.

This study aimed to investigate whether the plant species Stachys byzantina produces bioactives with the potential to delay the skin ageing process and treat hyperpigmentation conditions. The antioxidant action was assessed by 2,2-diphenyl-1-picrylhydrazylradical scavenging, Griess reaction, oxygen radical absorption capacity, and β-carotene bleaching assays. Inhibitory activities for tyrosinase, hyaluronidase, and elastase enzymes were tested. The antiglycation activity, the sun protection factor (SPF), and the toxicity to skin cells by MTT(3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide)assay were also evaluated. The ethanolic extract of S. byzantina aerial parts and all fractions obtained by solvent partition inhibited the tyrosinase enzyme at different levels. The dichloromethane fraction (DF) demonstrated the highest inhibition (IC50 = 63.5±10.9 µg/ml). DF also inhibited the hyaluronidase enzyme with IC50 = 369±11.64 μg/ml and elastase by 40% at 500 μg/ml. This fraction showed prominent antioxidant and antiglycation activities, high SPF, and no cytotoxicity at concentrations lower than 50 μg/ml. The phenolic and flavonoid contents were 116.30±6.7 (mgTAE/g) and 66.38±13.5 (mgQE/g), respectively. Chlorogenic acid (23.54±2.46mg/g) and verbascoside (203.97±19.8mg/g) were identified and quantified. Stachys byzantina is a potential source of cosmetic and therapeutic ingredients to reduce hyperpigmentation and the impacts caused by free radicals, advanced glycation end products, and sun radiation in skin ageing.

From small‐scale studies to an encompassing view: Inhibiting inflammation and clinically relevant enzymes with various extracts of Primula vulgaris using in vitro and in silico techniques

AbstractThe genus Primula holds great importance as a source of traditional remedies in various folk medicine systems. In the present study, we investigated the chemical composition and biological properties of different extracts (ethyl acetate, ethanol, ethanol/water, and water) of aerial parts and rhizomes of Primula vulgaris. To determine the chemical profile, the extracts were analyzed using ultrahigh performance liquid chromatography‐high‐resolution mass spectrometry (UHPLC‐HRMS) technique and flavonoids were a major group in this profile. The antioxidant capacity was demonstrated by in vitro chemical tests and in general the ethanol/water extract was found to be the most potent. Enzyme inhibition was studied against various enzymes and ethanol and ethanol/water extracts were more active than others. To assess the anti‐inflammatory potential of the extracts at the molecular level, human dermal fibroblasts (HDF) were treated with lipopolysaccharide (LPS). In vitro experiments showed that the levels of nuclear factor kappa B (NFKB), receptor for advanced glycation endproducts (RAGE), activator protein‐1 (AP‐1), interleukin 6 (IL‐6), interleukin 17 (IL‐17) and tumor necrosis factor (TNF)‐alpha were significantly reduced after treatment with the tested extracts. In addition, the extracts showed an inhibitory effect on the enzymes collagenase, elastase, and hyaluronidase, which are involved in the inflammatory process and destabilization of the extracellular matrix (ECM). P. vulgaris has been observed to modulate matrix metalloproteinase (MMP) synthesis by decreasing the concentration of cellular reactive oxygen species (ROS) during LPS‐induced inflammation. This study also examined the molecular binding and dynamic behavior of various enzymes and proteins associated with skin and infections using ligands derived from the tested extracts. Simulations with AutoDock Vina V1.1.2 and GROMACS 2023.1 showed that isoquercetin in particular showed superior performance in interactions with hyaluronidase. These findings are important for the development of potential therapeutic strategies for skin health and infection control. In summary, P. vulgaris can be considered as an important source of natural bioactive compounds for the development of effective health‐promoting applications in nutraceuticals, pharmaceuticals, and cosmetics

Multi-step synthesis, kinetics and in silico explorations of indole-Phenyl-1,2,4-triazole Bi-heterocyclic hybrids unified with 3-substituted benzamides as elastase inhibitors

In the present research work, a library of unique indole-phenyltriazole hybrids comprising 3-substituted-benzamide moiety was synthesized through convergent multi-step strategy. The structural confirmation of all synthesized molecules was corroborated by IR, 1H NMR, 13C MMR, EI-MS and CHN analysis data. The results of in vitro inhibitory potential of novel benzamides against elastase enzyme revealed that all molecules were potent inhibitors and 10d was most active compound among them having IC50 value of (0.197 ± 0.027 µM), relative to the standard (13.421 ± 0.016 µM). The Kinetics mechanism analyzed by Lineweaver-Burk plots which exposed that 10d inhibited this enzyme competitively by forming an enzyme-inhibitor complex. The inhibition constant Ki determined from Dixon plot for compound 10d was 0.85 µM. Moreover, cytotoxicity of these compounds was also profiled by hemolytic activity and it was observed that almost all these benzamides compounds displayed low cytotoxicity. These molecules also exhibited mild cytotoxicity toward red blood cell membrane. In addition, the in silico computational explorations fully supported the in vitro enzyme inhibitory results. The binding energy ranges from -7.1 to -9.1 kcal/mol, which showed that compound possessed good interaction tendencies to the pancreatic elastase target protein. So, it was anticipated from the experimental results and computational investigations of the current research that these derivatives might lead to further research gateways for obtaining better and safe nontoxic medicinal scaffolds for dealing with the elastase related ailments such as lungs diseases, pruritic skin disease and liver infection.

In vitro and in silico therapeutic properties of Artepillin C and Aromadendrin as collagenase and elastase inhibitors and investigation of anti-Ovarian cancer effects and antioxidant potential

In this investigation, the enzymes collagenase and elastase were inhibited by artepillin.C and aromadendrin molecules with excellent to good IC50 values of 16.65, and 0.79 μM for artepillin.C and 7.31, 19.38, and 10.56 μM for aromadendrin. Using the molecular modeling study, the chemical activity of these compounds against the enzymes, collagenase, and elastase were evaluated. The anti-cancer properties of the compounds were evaluated using the following cell lines: NIH: OVCAR-3, ES-2, UACC-1598, Hs 832(C).T [Hs832.Tc], TOV-21G, UWB1.289. The DPPH (1,1-diphenyl-2-pricrylhydrazyl) free radical scavenging assay was used to measure antioxidant activity, and the spectrophotometric method was used in this work. The chemical activities of artepillin.C and aromadendrin against collagenase and elastase were investigated utilizing the molecular docking study. The anti-cancer activities of the compounds were evaluated against NIH: OVCAR-3, ES-2, UACC-1598, Hs 832(C).T [Hs832.Tc], TOV-21G, UWB1.289 cell lines. Also, some cell lines had best results for aromadendrin like NIH: OVCAR-3, ES-2, UACC-1598, Hs 832(C).T [Hs832.Tc], TOV-21G, UWB1.289 (10.54 ± 0.94, 22.11 ± 2.52, 31.85 ± 4.73, 8.14 ± 1.52, 17.94 ± 1.88, and 24.31 ± 2.64 μM). The chemical activities of artepillin.C and aromadendrin against some of the expressed surface receptor proteins (folate receptor, CD44, EGFR, Formyl Peptide Receptor–Like 1, M2 muscarinic receptor, and estrogen receptors) in the mentioned cell lines were assessed using the molecular docking calculations. The outcomes provided information on potential interactions and their atomic-level properties. According to the docking scores, the compounds show a high affinity for binding to proteins and enzymes. Furthermore, these substances made good contact with the receptors and enzymes. As a result, these substances may have the ability to suppress cancer cells and enzymes.

Revealing the Potential of Chios Mastic Gum and Its Constituents for Cosmetic Applications through Chemical Profiling and Biological Evaluation

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.

ESSENTIAL OIL CONTENTS AND BIOLOGICAL ACTIVITIES OF THYMUS CANOVIRIDIS JALAS AND THYMUS SIPYLEUS BOISS.

Objective: Members of the Lamiaceae family are considered to be major sources of bioactive therapeutic agents. Many of them are important medicinal and aromatic plants used in traditional and modern medicine and in the food, cosmetic and pharmaceutical industries. The aim of this study was to investigate in detail the biological activities and chemical composition of the essential oils of Thymus canoviridis Jalas and Thymus sipyleus Boiss. species belonging to the genus Thymus, one of the most important genera of the Lamiaceae family. Material and Method: The essential oil content of the species was determined by GC-MS. Antioxidant activities of the essential oils were determined using lipid peroxidation, DPPH free radical, ABTS cation radical and CUPRAC methods. In addition, cytotoxic activities against breast cancer (MCF-7) and colon cancer (HT-29) cell lines and anticholinesterase (against AChE and BChE enzymes), urease, tyrosinase, elastase, collagenase and angiotensin converting enzyme inhibition activities were determined. Result and Discussion: When the essential oil composition of T. sipyleus was analyzed, the major compounds were 1,8-cineole (eucalyptol) (18.16%), camphor (15.08%) and endo-borneol (11.63%), while T. canoviridis was found to be rich in carvacrol (72.88%). T. canoviridis showed high antioxidant activity in lipid peroxidation (IC50: 45.72±0.12 μg/ml), ABTS (IC50: 6.12±0.03 μg/ml) and CUPRAC (IC50: 5.31±0.01 μg/ml) methods. The selectivities of T. canoviridis and T. sipyleus species against MCF-7 cell line were 4.39 and 6.81, respectively. In the enzyme inhibition studies, both Thymus species showed moderate inhibition activity against BChE enzyme (Inhibition%: 57.88±1.14, 39.21±0.89, respectively). In addition, T. sipyleus showed moderate inhibition of elastase enzyme (Inhibition%: 25.33±0.79). When the results are evaluated in general, it can be said that T. canoviridis essential oil with its rich carvacrol content and high cytotoxic and antioxidant activity can be preferred as a safer and natural option instead of synthetic preservatives in food, pharmaceutical and cosmetic industries to extend shelf life and ensure food safety.

(Chemical) Roles of HOCl in Rheumatic Diseases.

Chronic rheumatic diseases such as rheumatoid arthritis (RA) are characterized by a dysregulated immune response and persistent inflammation. The large number of neutrophilic granulocytes in the synovial fluid (SF) from RA patients leads to elevated enzyme activities, for example, from myeloperoxidase (MPO) and elastase. Hypochlorous acid (HOCl), as the most important MPO-derived product, is a strong reactive oxygen species (ROS) and known to be involved in the processes of cartilage destruction (particularly regarding the glycosaminoglycans). This review will discuss open questions about the contribution of HOCl in RA in order to improve the understanding of oxidative tissue damaging. First, the (chemical) composition of articular cartilage and SF and the mechanisms of cartilage degradation will be discussed. Afterwards, the products released by neutrophils during inflammation will be summarized and their effects towards the individual, most abundant cartilage compounds (collagen, proteoglycans) and selected cellular components (lipids, DNA) discussed. New developments about neutrophil extracellular traps (NETs) and the use of antioxidants as drugs will be outlined, too. Finally, we will try to estimate the effects induced by these different agents and their contributions in RA.

Green synthesis of silver nanoparticles using food supplement from Avena sativa L., and their antioxidant, antiglycation, and anti-aging activities: In vitro and in silico studies

Food supplement of Avena sativa L. dry extract (AsDE) is produced with green leaves of wild oats and it is commonly used in patients with cognitive disorders and muscle deficits. These symptoms and pathologies can be promoted and/or aggravated by oxidative stress and protein glycation. In order to prevent or treat these disorders, new formulations that could enhance and benefit AsDE have been developed and the formation of metallic nanoparticles by green synthesis has shown to be an alternative. Thus, this study aimed to prepare silver nanoparticles (AgNPs) by green synthesis using AsDE and characterize these AgNPs. Besides, it aimed to evaluate in vitro antioxidant and antiglycation activities of AsDE and AgNPS, and analyze in vitro and in silico inhibitory action on aging enzymes (collagenase, elastase, and tyrosinase). UV–visible spectroscopy, Zeta potential and scanning transmission electron microscopy were used to characterize AgNPs. Antioxidant activity was determined by DPPH free radical scavenging, ferric ion reducing power (FRAP), lipid peroxidation inhibition, ABTS radical scavenging, and oxygen radical absorption capacity (ORAC). Molecular docking analyses were performed to evaluate interactions between the major compounds of AsDE and aging enzymes or DNA. Antioxidant tests demonstrated antioxidant activity of 19.67 % for AsDE and 42.98 % for AgNPs in DPPH test; and 115.67 µM Trolox Equivalent (TE)/g for AsDE and 554.33 µM TE/g for AgNPs in FRAP test. In addition, AsDE and AgNPs inhibited 25.40 % and 11.33 % of lipid peroxidation, respectively, and presented 255.22 µM TE/g for AsDE and 317.44 µM TE/g for AgNPs in ABTS test. AsDE and AgNPs exhibited antioxidant potential observed in ORAC assay. In addition, AsDE presented inhibitory action on collagenase, elastase and tyrosinase activities (78.23 %, 68.14 % and 62.27 % inhibition, respectively). Antiglycation tests demonstrated that bovine serum albumin exposed to ribose and treated with AsDE at 0.5 mg/mL exhibited the highest percentage of free amino groups (45.21 %), while samples treated with AgNPs showed 35.41 % free amino groups. On the other hand, AgNPs exhibited the highest percentage of inhibition of advanced glycation end-products formation (85.75 %), which did not differ from aminoguanidine, a known antiglycation agent. Besides, relative electrophoretic mobility assay demonstrated that samples treated with AsDE at 0.1 mg/mL or AgNPs showed antiglycation activity comparable to aminoguanidine. The in silico assays showed interactions between the major compounds of AsDE and aging enzymes or DNA. Therefore, results represent an important indicator for the development and discovery of new nanostructured pharmaceutical and cosmetic formulations using plants and their bioactive compounds.

γ-Mangosteen, an autophagy enhancer, prevents skin-aging via activating KEAP1/NRF2 signaling and downregulating MAPKs/AP-1/NF-κB-mediated MMPs

BackgroundMangosteens, a naturally occurring xanthones, found abundantly in mangosteen fruits. The anti-skin aging potential of γ-mangosteen (GM) remains unexplored; therefore, we investigated the UVB-induced anti-skin aging of GM via activation of autophagy. HypothesisWe hypothesized that GM exerts antioxidant and anti-aging capabilities both in vitro and in vivo through activation of autophagy as well as control of KEAP1/NRF2 signaling and MAPKs/AP-1/NF-κB-mediated MMPs pathways. MethodsThe anti-skin aging effects of GM were studied using HDF cells and a mice model. Various assays, such as DPPH, ABTS, CUPRAC, FRAP, and ROS generation, assessed antioxidant activities. Kits measured antioxidant enzymes, SA-β-gal staining, collagen, MDA content, si-RNA experiments, and promoter assays. Western blotting evaluated protein levels of c-Jun, c-Fos, p-IκBα/β, p-NF-κB, MAPK, MMPs, collagenase, elastin, KEAP1, NRF2, HO-1, and autophagy-related proteins. ResultsGM exhibited strong antioxidant, collagenase and elastase enzyme inhibition activity surpassing α- and β-mangosteen. GM competitively inhibited elastase with a Ki value of 29.04 µM. GM orchestrated the KEAP1-NRF2 pathway, enhancing HO-1 expression, and suppressed UVB-induced ROS in HDF cells. NRF2 knockdown compromised GM's antioxidant efficacy, leading to uncontrolled ROS post-UVB. GM bolstered endogenous antioxidants, curbing lipid peroxidation in UVB-exposed HDF cells and BALB/c mice. GM effectively halted UVB-induced cell senescence, and reduced MMP-1/-9, while elevated TIMP-1 levels, augmented COL1A1, ELN, and HAS-2 expression in vitro and in vivo. Additionally, it suppressed UVB-induced MAPKs, AP-1, NF-κB phosphorylation. Pharmacological inhibitors synergistically enhanced GM's anti-skin aging potential. Moreover, GM inhibited UVB-induced mTOR activation, upregulated LC3-II, Atg5, Beclin 1, and reduced p62 in both UVB induced HDF cells and BALB/c mice, while blocking of autophagy successfully halt the GM effects against the UVB-induced increase of cell senescence, degradation of collagen through upregulation of MMP-1, underscoring GM's substantial anti-skin aging impact via autophagy induction in vitro and in vivo. ConclusionTogether, GM has potent antioxidant and anti-skin aging ingredients that can be used to formulate skin care products for both the nutraceutical and cosmeceutical industries.

The investigation of the interaction determination between carbendazim and elastase, using both in vitro and in silico methods

Pesticides, including fungicides, are one of the important groups of environmental toxins that affect human and animal health. Studies have shown that these compounds are considered chemical pollutants. Carbendazim is a systemic fungicide. Unfortunately, excessive use of carbendazim has caused environmental pollution all over the world. In this study, the effect of carbendazim on the enzyme elastase (secreted from the endocrine gland of the pancreas) has been investigated. In a study, the performance and reaction of carbendazim with elastase were investigated using spectroscopic techniques. The stability and structure of elastase enzymes were studied under the influence of carbendazim. The results of fluorescence emission and UV–visible absorption spectrum showed that in the presence of carbendazim, there is an increase in UV–Vis absorption and a decrease in the intensity of the intrinsic fluorescence emission in the protein spectrum. Additionally, a decrease in the thermal stability of elastase was observed in the presence of carbendazim. The stability and structure of elastase enzyme were investigated in the presence of carbendazim. The results revealed that the UV–Vis absorption increased due to the presence of carbendazim, as indicated by the hyperchromic spectrum at 220 and 280 nm peaks. Additionally, the intrinsic fluorescence emission in the protein spectrum decreased with increasing carbendazim concentration at three different temperatures (298, 303, and 313 K). Moreover, the study demonstrated that the TM decreased from 2.59 to 4.58 with the increase of carbendazim, suggesting a decrease in the stability of the elastase structure in response to the elevated carbendazim concentration. According to the results of the research, the interaction between elastase and carbendazim has occurred, and changes have been made in the enzyme under the influence of carbendazim. The formation of the complex between elastase and carbendazim was consistent with the results obtained from molecular simulation and confirmed the thermodynamic data.

Cubeb (Piper cubeba L.): nutritional value, phytochemical profiling and dermacosmeceutical properties.

Cubeb, Piper cubeba L., has been used for centuries in traditional medicine and culinary practices, with a wide range of biological and pharmacological activities. Herein, we determined the phytochemical profile, mineral, fatty acids, and amino acid contents of P. cubeba berries and assessed the dermacosmeceutical properties of their water extract and essential oil (EO). These included assessing their antioxidant and antibacterial activities as well as their in vitro inhibitory activities against tyrosinase and elastase enzymes. In addition, molecular docking and molecular dynamics studies were performed on the major identified compounds of the EO. A total of forty-three compounds belonging to organic acids, phenolic acids and flavonoids were found in the water extract, while 36 volatile compounds were identified in the EO with Z-isoeugenol, dihydroeugenol, β-pinene, E-caryophyllene, and 1,8-cineole as major constituents. The berries were found to be rich in sodium and iron, have moderate zinc content along with low contents of total nitrogen, phosphorus, and potassium. Amino acid analysis revealed a considerable concentration of isoleucine and phenylalanine, whereas 11,14,17-eicosatrienoic acid and linoleic acid were identified as the major fatty acids. In the DPPH and FRAP assays, the water extract elicited considerable antioxidant activity compared to the reference compounds. Enzyme inhibitory assays revealed that the EO had a potential to inhibit tyrosinase and elastase enzymes with IC50 values of 340.56 and 86.04 μg/mL, respectively. The water extract and EO completely inhibited the bacterial growth at MIC of 50 mg/mL and 20%, respectively. At sub-MIC concentrations, the extract and the EO substantially reduced the biofilm formation by up to 26.63 and 77.77%, respectively, as well as the swimming and swarming motilities in a dose-dependent manner. Molecular docking and molecular dynamics showed that the five main components of P. cubeba EO could be the major contributors to the elastase and tyrosinase inhibitory effect. This study emphasizes the promising potential of P. cubeba as a valuable source of natural compounds that can be utilized for the development of innovative pharmaceuticals, dietary supplements, and dermacosmeceutical agents.

Evaluation of potential anti-aging effects of Achillea phrygia Boiss. & Balansa (Asteraceae)

This study aims to determine the anti-aging effects of Achillea phrygia, an endemic plant, by evaluating its sun protection factor (SPF) level, antioxidant activity, total phenolic content, extracellular matrix-degrading enzymes (ECM) inhibition, genotoxic/anti-genotoxic, and cytotoxic activities. The SPF level was assessed using an in vitro quantitative method, while antioxidant capacity was determined through DPPH, β-carotene, and hydroxyl-radical (H2O2) scavenging assays. The total phenolic content was quantitatively conducted using the Folin Ciocalteu reagent. The inhibition of ECM-degrading enzymes was determined using matrix metalloproteinase-1 (MMP-1), hyaluronidase, and elastase enzymes. Genotoxic/anti-genotoxic properties were assessed using the AMES Salmonella/microsome assay, and cytotoxicity effects were assessed through the MTT assay. The results indicated that A. phrygia showed moderate SPF activity (SPF = 4.013) and exhibited IC50 values of 0.183 ± 0.03, 0.079 ± 0.51, and 1.18 ± 0.35 mg/mL for DPPH, β-carotene, and hydroxyl-radicals, respectively. The total phenolic content was measured to be 23.56 ± 1.42 mg GAE/g dry extract. Furthermore, the extract demonstrated inhibition of MMP-1 (47.98%) and elastase (39.2%) activities. Importantly, it did not induce DNA damage and showed antigenotoxic activity ranging from 10% to 65.6%. The cytotoxicity assay revealed an IC50 value of 42.41±4.05 µg/mL. These findings suggest that A. phrygia could be utilized as a cosmetic ingredient in skincare products due to its ability to protect against UV radiation, exhibit antioxidant properties, prevent extracellular matrix degradation, and inhibit DNA damage.

Neutrophil extracellular traps promote invasion and metastasis via NLRP3-mediated oral squamous cell carcinoma pyroptosis inhibition

Neutrophil extracellular traps (NETs) are reticular structures composed of neutrophil elastase (NE), cathepsin G (CG) and DNA-histone enzyme complexes. Accumulating evidence has revealed that NETs play important roles in tumor progression, metastasis, and thrombosis. However, our understanding of its clinical value and mechanism of action in oral squamous cell carcinoma (OSCC) is limited and has not yet been systematically described. Here, we aimed to investigate the clinical significance of NETs in OSCC and the mechanisms by which they affect its invasive and metastatic capacity. Our results demonstrated that high enrichment of NETs is associated with poor prognosis in OSCC, and mechanistic studies have shown that NE in NETs promotes invasion and metastasis via NLRP3-mediated inhibition of pyroptosis in OSCC. These findings may provide a new therapeutic approach for OSCC.

Assessing salivary innate immune responses in adolescents with dentofacial abnormalities and central precocious puberty

Relevance. Central precocious puberty (CPP), characterized by a high incidence of 60% among endocrinopathies during puberty, significantly influences dentofacial development and the local immune defense mechanisms in adolescents. However, the impact on saliva's innate immune function remains underexplored.This study aims to evaluate the innate immune function of saliva in individuals with CPP coexisting with dentofacial abnormalities.Materials and methods. An analysis of saliva biochemical markers was conducted on 59 adolescents diagnosed with CPP and dentofacial abnormalities, alongside a control group of 21 age-matched individuals with dentofacial abnormalities but without systemic conditions, aged 13-18 years. The evaluation focused on lysozyme and urease levels, dysbiosis markers indicating oral microbiota status, malondialdehyde (MDA) levels, and catalase activity to gauge the balance between pro-oxidant and antioxidant systems, alongside proteolytic enzyme elastase activity.Results. Adolescents with combined CPP and dentofacial abnormalities exhibited weakened antioxidant and antibacterial defenses, evidenced by decreased catalase activity (0.105 ± 0.020 mkat/l, p < 0.001) and lysozyme levels (54.7 ± 3.2 U/l, p < 0.02), against a backdrop of increased lipoperoxidation and microbial contamination (MDA levels at 0.48 ± 0.11 μmol/l, p < 0.05, urease levels at 0.713 ± 0.015 μkat/l, p < 0.01), and heightened inflammation (elastase activity up to 2.71 ± 0.12 μkat/l, p < 0.01).Conclusion. The diminished innate immune function in saliva necessitates the development of strategies to enhance it, serving as a preventive measure against gingivitis in patients with CPP.

In vitro Evaluation of Skin Related Enzyme Inhibitory Effect and Emulgel Formulation Development Studies of Onobrychis Argyrea subsp. Argyrea with Phytochemical Analysis.

Species of Onobrychis have been used to treat skin disorders such as wounds and cuts in folk medicine and Onobrychis argyrea subsp. argyrea (OA) commonly known as 'silvery sainfoin', is a member of this genus. In this study, it was aimed to investigate the skin-related biological activities and phytochemical characterization of OA. Moreover, an emulgel formulation was developed from the main methanolic extract of the plant (OAM). Initially, to identifiy of the active fractions, aerial parts of the plant material was extracted with methanol and fractionated by n-hexane, chloroform, ethyl acetate and n-butanol, respectively. Antioxidant activity was determined by CUPRAC, TOAC, FRAP and DPPH assays. Thereafter, the inhibition potential of OAM, novel formulation and all fractions was measured against elastase, collagenase, tyrosinase and hyaluronidase enzymes. OAM was analyzed and characterized by LC/MS-MS. The major bioactive flavonoids which are rutin and isoquercetin were measured and compared as qualitative and quantitative via high performance thin layer chromatography (HPTLC) analysis in OAM and fractions. The results showed that extracts of OA can be a potential cosmeceutical agent for skin related problems.

From in silico to in vitro: Hydrolysis of cheese whey to obtain peptides with anti-aging and anti-hypertensive activities

Our goal was to identify potential anti-aging and anti-hypertensive peptides from the most abundant protein in cheese whey (CW), β-lactoglobulin (BLG), using a multidisciplinary approach. These peptides were generated by three different proteases: trypsin, chymotrypsin and subtilisin. Our bioinformatics approach indicated that trypsin or subtilisin are good candidates for generating anti-hypertensive peptides, whereas chymotrypsin has the highest potential to generate anti-aging fragments, especially anti-elastase. This is mainly due to the presence of chymotryptic peptides with highly similar structures to elastase inhibitors. In addition, we aimed to perform the enzymatic hydrolysis of BLG and evaluate in vitro anti-aging and anti-hypertensive potential by assessing the inhibition to collagenase, elastase and angiotensin-converting enzyme (ACE). We found that chymotrypsin was able to generate a <10 kDa fraction of BLG-hydrolysate that reduces elastase activity (∼15%), which corroborates our interpretation of the in silico data. This fraction also showed reduced ACE activity (∼18%), though statistically similar to the intact protein (∼16%). Peptidomic analysis revealed that peptide sequences with predicted anti-elastase activity were present both in BLG and CW chymotrypsin-hydrolyzed samples, though at higher concentrations in the isolated protein, which may support our experimental results. Lastly, CW was hydrolyzed by chymotrypsin and spray-dried to be used as ingredient in a potentially functional milk-based drink. Hydrolysis significantly impacted the characteristics of the dried powder and the drink containing CW hydrolysate attained better evaluation in sensory analysis. Thus, enzymatic hydrolysis with chymotrypsin generated a potentially functional product that improves bioactivity and functionality of CW.

Exploration of newly synthesized amantadine-thiourea conjugates for their DNA binding, anti-elastase, and anti-glioma potentials

Three newly synthesized amantadine thiourea conjugates namely MS-1 N-(((3 s,5 s,7 s)-adamantan-1-yl)carbamothioyl)benzamide, MS-2 N-(((3 s,5 s,7 s)-adamantan-1-yl)carbamothioyl)-4-methylbenzamide and MS-3 N-((3 s,5 s,7 s)-adamantan-1-ylcarbamothioyl)-4-chlorobenzamide were investigated for their structures, bindings (DNA/ elastase), and for their impact on healthy and cancerous cells. Theoretical (DFT/docking) and experimental {UV–visible (UV-), fluorescence (Flu-), and cyclic voltammetry (CV)} studies indicated binding interactions of each conjugate with DNA and elastase enzyme. Theoretically and experimentally calculated binding parameters for conjugate – DNA interaction revealed MS-3 – DNA to have most significant binding with comparatively greater values of binding parameters {(Kb/M−1: docking, 3.8 × 105; UV-, 5.95 × 103; Flu-,1.55 × 105; CV, 1.52 × 104), (∆G/ kJmol−1: docking, −32.09; UV-, −22.40; Flu-,-30.81; CV, −24.82)}. The docked structures, greater bindings site size values (n), and the trend in DNA viscosity changes in the presence of each conjugate concentration confirmed a mixed binding mode of interaction among them. Conjugate – elastase binding by docking agreed with the experimental anti-elastase findings. Cytotoxicity studies of each tested conjugate demonstrated greater cytotoxicity for cancerous (MG-U87) cells in comparison to control, while for the normal (HEK-293) cells the cytotoxicity was found comparatively low. Overall exploration suggested that MS-3 is the most effective candidate for DNA binding, anti-elastase, and for anti-glioma activities.

EVALUATION OF THE ANTI-TYROSINASE-ANTI-AGING POTENTIAL AND METABOLITE PROFILING FROM THE BIOACTIVE FRACTION OF CORN COB (ZEA MAYS L.)

Objective: Maize (Zea mays L.) is a crop that has been widely cultivated in Indonesia. Using corn kernels on a large scale will produce much corn cob waste, usually unused. According to the literature search, corn cobs’ phytochemical studies and pharmacological activities still need to be improved. This study aims to determine the content of secondary metabolites (metabolite profiling) and their antityrosinase and anti-aging potential.&#x0D; Methods: Corn cobs were macerated with methanol and fractionated with n-hexane, ethyl acetate, and butanol. The phytochemical profiling approach of the methanol extract was performed by liquid chromatography-mass spectra (LC-MS/MS). Anti-tyrosinase and anti-aging bioactivity were evaluated by thin layer chromatography (TLC)-bioautography and IC50 spectrophotometrically.&#x0D; Results: The evaluation results show that the butanol fraction leads to a potential value (IC50 99.92 µg/ml). Several compounds, especially flavonoid compounds (including catechin; kaempferol 3-arabinofuranoside 7-rhamnoside; 6,8-Di-C-beta-D-arabino pyranosyl apigenin; 5,7-Dihydroxy-8,4’-dimethoxyisoflavone) were identified by LC-MS/MS by comparing the molecular mass of MS/MS data with literature data.&#x0D; Conclusion: Based on this study, it can be concluded that butanol is the fraction that most actively inhibits tyrosinase, elastase, and collagenase enzymes, which means it potentially becomes a new anti-aging candidate.

Content analysis, nanoparticle forming potential and acaricidal effects on Tick (Hyalomma marginatum) of essential oils from two common Thymbra species

In this study, the fatty acid, essential oil and aroma composition of Thymbra sintenisii Bornm. & Aznav. subsp. sintenisii Bornm. & Aznav (TSS) and Thymbra sintenisii Bornm. & Azn. subsp. isaurica P.H.Davis (TSI) were determined by GC-MS. The cytotoxic activity was determined with MTT assay and antioxidant activities were determined with DPPH, ABTS, CUPRAC methods. Urease, acetyl, butyryl, angiotensin, α-amylase, tyrosinase, elastase, and collagenase enzyme inhibition activities of these species were also determined. The acaricidal effects of essential oils from aerial parts of TSS and TSI on ticks and the nanoparticle formation potential of their essential oils with AgNO3 were determined and compared. The main components of TSS essential oil and aroma were determined as carvacrol and o-cemen (66.78 and 35.44%, respectively), the major components of TSI essential oil and aroma were carvacrol and γ-terpinene (63.78 and 31.44%, respectively). The species exhibited remarkably high antioxidant activity, particularly in the ABTS and CUPRAC methods. It was also determined that the butyrylcholinesterase and acetylcholinesterase enzyme inhibition activities (Inhibition%: 76.12±0.83 and 74.13±0.33, respectively) of the species were quite high. Moderate antimicrobial activity against the bacteria was observed for both essential oils, with MIC values ranging from 312 to 1250 μg/mL. Moreover, essential oils of these species showed good acaricidal activity on Hyalomma marginatum (LC50: 11.82 μg/mL for TSS; LC50: 57.64 μg/mL for TSI). The high antioxidant, anticholinesterase, and acaracidal activities observed in both species are attributed to the presence of carvacrol in their essential oils. The species have potential to be used in food supplements-preservatives, pharmaceutical industry.