Elastase Inhibitor Research Articles

Enzymatic synthesis of a skin active ingredient - glochidone by 3-ketosteroid dehydrogenase from Sterolibacterium denitrificans

In this study, we applied AcmB2, sourced from Sterolibacterium denitrificans, to catalyze the oxidative dehydrogenation of 3-ketolupeol (lupenone), a derivative of lupeol, triterpene obtained from birch bark. This enzymatic Δ1-dehydrogenation catalyzed by AcmB2 yielded glochidone, a bioactive compound frequently obtained from medicinal plants like Salvia trichoclada and Maytenus boria. Glochidone is known for its broad biological activities, including antibacterial, antifungal, anti-inflammatory, anticancer, antidiabetic as well as acetylcholinesterase inhibition. Our research demonstrates >99% conversion efficiency with 100% regioselectivity of the reaction. The effective conversion to glochidone employed an electron acceptor e.g., potassium hexacyanoferrate III, in mild, environmentally friendly conditions: 8–16% 2-hydroxypropyl-β-cyclodextrin, and 2–3% 2-methoxyethanol. AcmB2 reaction optimum was determined at pH 8.0 and 30 °C. Enzyme's biochemical attributes such as electron acceptor type, concentration and steroid substrate specificity were investigated. Among 4-, 5- and 6-ring steroid derivatives androst-4-en-3,17-dione and testosterone propionate were determined as the best substrates of AcmB2. Δ1-Dehydrogenation of substrates such as lupenone, diosgenone and 3-ketopetromyzonol was confirmed. We have assessed the antioxidant and rejuvenating characteristics of glochidone as an active component in formulations, considering its precursors, lupeol, and lupenone as well. Glochidone exhibited limited antioxidant and chelating capabilities compared to lupeol and reference compounds. However, it demonstrated robust rejuvenating properties, with a sirtuin induction level of 61.5 ± 1.87%, notably surpassing that of the reference substance, E-resveratrol (45.15 ± 0.09%). Additionally, glochidone displayed 26.5±0.67 and 19.41±0.76% inhibition of elastase and collagenase, respectively. The safety of all studied triterpenes was confirmed on skin reconstructed human Epidermis model. These findings provide valuable insights into the potential applications of glochidone in formulations aimed at addressing skin health concerns. This research presents the first example of an enzyme in the 3-ketosteroid dehydrogenase (KstD) family catalyzing the Δ1-dehydrogenation of a pentacyclic triterpene. We also explored structural differences between AcmB, AcmB2, and related KstDs pointing to G52 and P532 as potentially responsible for the unique substrate specificity of AcmB2. Our findings not only highlight the enzyme's capabilities but also present novel enzymatic pathways for bioactive compound synthesis.

The potential of Paeonia lactiflora pall seeds oil as a pure natural cosmetics raw material: InVitro findings.

As a traditional Chinese herbal medicine, Paeonia lactiflora Pall is rich in various active ingredients such as polysaccharides and total flavonoids while having ornamental value. It has potential application value in the development of food and cosmetics. To study the invitro efficacy of Paeonia lactiflora Pall seeds oil. Firstly, the levels of linolenic acid and linoleic acid in Paeonia lactiflora Pall seeds oil were quantified using gas chromatography. The impact of Paeonia lactiflora Pall seeds oil on the proliferation rate of B16F10 cells was assessed through the CCK-8 method, while the melanin content of B16F10 cells was determined using the sodium hydroxide lysis method. The inhibitory effects of Paeonia lactiflora Pall seeds oil on elastase, collagenase and hyaluronidase were evaluated by biochemical techniques invitro. Lastly, the hen's egg chorioallantoic membrane test (HET-CAM) was conducted to confirm the absence of eye irritation caused by Paeonia lactiflora Pall seeds oil. Paeonia lactiflora Pall seeds oil within a certain volume concentration range (0.5%-4%) had no effect on the proliferation of B16F10 cells. Paeonia lactiflora Pall seeds oil showed significant inhibition of elastase, collagenase and hyaluronidase. Notably, the highest concentration tested, 4% Paeonia lactiflora Pall seed oil, yielded the most pronounced outcomes without causing any irritation. A certain concentration of Paeonia lactiflora Pall seeds oil has a significant effect on decreasing the melanin content in B16F10 cells and inhibiting the activities of elastase, collagenase, and hyaluronidase, which can provide a reference for the development of pure natural cosmetics raw materials.

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.

Targeted Library of Phosphonic-Type Inhibitors of Human Neutrophil Elastase.

Despite many years of research, human neutrophil elastase (HNE) still remains an area of interest for many researchers. This multifunctional representative of neutrophil serine proteases is one of the most destructive enzymes found in the human body which can degrade most of the extracellular matrix. Overexpression or dysregulation of HNE may lead to the development of several inflammatory diseases. Previously, we presented the HNE inhibitor with kinact/KI value over 2,000,000 [M-1s-1]. In order to optimize its structure, over 100 novel tripeptidyl derivatives of α-aminoalkylphosphonate diaryl esters were synthesized, and their activity toward HNE was checked. To confirm the selectivity of the resultant compounds, several of the most active were additionally checked against the two other neutrophil proteases: proteinase 3 and cathepsin G. The developed modifications allowed us to obtain a compound with significantly increased inhibitory activity against human neutrophil elastase with high selectivity toward cathepsin G, but none toward proteinase 3.

Otras enfermedades hepáticas de causa genética, metabólica y endocrinológica

Liver diseases of genetic origin comprise a series of diseases that manifest as abnormalities in liver structure and/or function due to genetic mutations. This update will discuss some of them, such as alpha-1 antitrypsin deficiency (A1AT), Wilson's disease, lysosomal acid lipase deficiency (LAL-D), and other congenital diseases related to bile acid and bilirubin metabolism.Alpha-1 antitrypsin deficiency is an autosomal codominant condition characterized by a decrease in secretion of alpha-1 antitrypsin protein, which acts as an inhibitor of neutrophil elastase. This results in liver and lung damage due to an accumulation of misfolded protein in hepatocytes and uncontrolled elastase activity. On the other hand, in Wilson's disease, an autosomal recessive disorder, mutations in the ATP7B gene impair copper metabolism. This results in an accumulation of toxic copper in the liver and other organs, causing hepatitis, cirrhosis, and neuropsychiatric manifestations. Lysosomal acid lipase deficiency is an autosomal recessive disorder caused by mutations in the LIPA gene. This results in the accumulation of cholesterol and triglyceride esters in lysosomes, leading to hepatomegaly, hyperlipidemia, and premature atherosclerosis. Finally, the set of diseases related to bilirubin and bile acid metabolism involve abnormalities in the production, transport, and excretion of bilirubin and bile acids. They are associated with cholestasis and hepatocellular damage, which can lead to liver failure and death.

NETworking for Health and in Disease: Neutrophil Extracellular Traps in Pediatric Surgical Care.

This comprehensive review examines the role of Neutrophil Extracellular Traps (NETs) in pediatric surgery. Focusing on NET formation, functions, and implications, this study highlights their dual impact in infection control and contribution to tissue damage after surgery. It covers the role of NET formation in a range of pediatric conditions including immunothrombosis, formation of peritoneal adhesions, appendicitis, burns, gallstones, tumors, and necrotizing enterocolitis (NEC). The results underscore the significance of NETs in fighting infections and their association with complications like sepsis and delayed wound healing. The breakdown products of NETs as a diagnostic tool of the clinical course of acute appendicitis will also be discussed. Understanding NET formation in the pathophysiology can potentially help to find new therapeutic approaches such as the application of DNase and elastase inhibitors to change the clinical course of various diseases in pediatric surgery such as improvement of wound healing, adhesion formation, NEC, and many more.

Characterization of extracellular trap production and release by equine neutrophils in response to different stimuli

This study explores Neutrophil Extracellular Trap (NET) formation in equine neutrophils, which is crucial for eliminating infections and is implicated in various equine inflammatory diseases. We investigated the molecular pathways involved in NET release by equine neutrophils in response to stimuli. We use PMA, A23187, LPS, PAF, OZ, and cytokines, observing NET release in response to PMA, PAF, and A23187. In contrast, LPS, OZ, and the cytokines tested did not induce DNA release or did not consistently induce citrullination of histone 4. Peptidyl-arginine deiminase inhibition completely halted NET release, while NADPH oxidase and mitochondrial reactive oxygen species only played a role in PMA-induced NETs. Neutrophil elastase inhibition modestly affected PAF-induced NET liberation but not in PMA or A23187-induced NET, while myeloperoxidase did not contribute to NET release. We expect to provide a foundation for future investigations into the role of NETs in equine health and disease and the search for potential therapeutic targets.

Skin Anti-Aging Potential through Whitening and Wrinkle Improvement Using Fermented Oil Derived from Hermetia illucens Larvae.

As the aging population increases, so has interest among emerging seniors in anti-aging ingredients that enhance functionality by incorporating fermentation with natural materials. In this study, fermentation conditions for enhancing the functionality of Hermetia illucens larvae oil (HIO) were established, and its anti-aging potential was evaluated. First, the lipase activity and amount of lipid degradation products of the fermentation strains were measured in order to select Lactobacillus gasseri and Lactiplantibacillus plantarum as the strains with high fermentation ability. A fermentation period of 28 d and a fermentation method that uses only the strain culture medium were established by evaluating the fermentation degree after fermenting HIO with the selected strains. The whitening functionality test results of fermented HIO (FHIO) showed an increase of approximately 20% in extracellular tyrosinase inhibition activity compared with HIO. Additionally, within melanocytes, there was a 12% increase in tyrosinase inhibition activity and a 26% enhancement in melanin production inhibition ability. For wrinkle-improving functionality, it was observed that, for fibroblasts, there was a 10% increase in collagen production, a 9% increase in collagenase inhibition ability, and an 8% increase in elastase inhibition ability. Therefore, FHIO was confirmed to be an effective cosmetic raw material, with high functionality for anti-aging within the senior generation. This is achieved through increased whitening and wrinkle-improving functionality.

Volatiles from French and Croatian Sea Fennel Ecotypes: Chemical Profiles and the Antioxidant, Antimicrobial and Antiageing Activity of Essential Oils and Hydrolates.

Sea fennel is a halophytic plant rich in valuable nutritional components and is characterized by pleasant organoleptic properties. While its essential oils (EOs) are well investigated, there are no reports on the volatiles from their corresponding hydrolates, which are the main by-products of EO isolation, as well as on their biological activity. Therefore, the composition and biological activities of EOs and corresponding hydrolates of sea fennel from Atlantic (French, FRA) and Mediterranean (Croatian, CRO) ecotypes were investigated and compared. The EO from the CRO sample was characterized by an abundance of sabinene and limonene, while that from the FRA ecotype was rich in dillapiol and carvacryl methyl ether. The CRO hydrolate was rich in terpinen-4-ol and 10-(acetylmethyl)-3-carene, while dillapiol, thymyl methyl ether and γ-terpinene were the main compounds in the FRA sea fennel hydrolate. The biological activities of the EOs and hydrolates were evaluated for their antioxidant (with DPPH, NO, FRAP and ORAC bioassays), antimicrobial (against some Gram+ and Gram- spoilage bacteria) and antiageing (tyrosinase, elastase and collagenase inhibition) activities. Both EOs showed low reducing powers and antiradical activities while the ability of both hydrolates to quench NO was slightly higher (35-39% if inhibition). The FRA EO showed low activity against Staphylococcus aureus (8 mm), while CRO moderately inhibited the growth of P. aeruginosa (8 mm), but strongly inhibited the other two bacterial strains. While the French EO showed no antityrosinase and anticollagenase activity, the Croatian oil significantly inhibited both enzymes (IC50 of 650 µg/mL and IC50 of 2570 µg/mL, respectively) probably due to the dominance of limonene and sabinene. Neither EO exhibited antielastase properties, while the hydrolates from both ecotypes showed no antiageing activity, regardless of the enzyme tested. The EOs from the aerial parts of sea fennel from FRA and CRO differed greatly in composition, resulting in different activities. The Croatian samples appeared to have better biological properties and are therefore good candidates for applications as preservatives or antiageing agents.

Cosmeceutical Activities of Essential Oils from the Rhizomes of Plants in the Zingiberaceae Family

The plants in the Zingiberaceae family are often rhizomatous annual or perennial herbs with considerable economic value. This study aimed to investigate the cosmeceutical activities of essential oils from the rhizomes of Hedychium coronarium J. Koenig, Curcuma caesia Roxb., and Curcuma longa L. The essential oils were obtained from hydrodistillation and subjected to investigations of antioxidant activities by 2,2-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric reducing antioxidant power, and ferric thiocyanate assay. Furthermore, the whitening properties were investigated in terms of anti-tyrosinase inhibition, whereas the anti-skin aging activities were investigated through collagenase, elastase, and hyaluronidase inhibition. The results showed that all essential oils exhibited low antioxidant, anti-elastase, and anti-collagenase activities. On the other hand, H. coronarium oil was the only essential oil in the present study, which was found to possess anti-tyrosinase activity, with the inhibition of 98.4 ± 5.5% and 30.2 ± 1.7% at the final concentration of 0.1 mg/ml, when the substrate were L-tyrosine and L-DOPA, respectively. Interestingly, it was highlighted that the anti-tyrosinase activity of H. coronarium oil was stronger than that of kojic acid, a well-known whitening agent, when the substrate was L-tyrosine. Besides, H. coronarium oil and C. longa oil exhibited promising anti-hyluronidase activities with inhibition of 83.6 ± 1.0% and 82.9 ± 2.2%, respectively, which were comparable to that of epigallocatechin gallate (100.0 ± 0.0%). All essential oil induced irritations in the HET-CAM test at the tested concentration of 5 mg/ml in 10% dimethyl sulfoxide in olive oil. Therefore, H. coronarium oil was suggested as an attractive ingredient in cosmeceutical products for whitening and anti-aging.

Nanoemulsions Containing Mucuna pruriens (L.) DC. Seed Extract for Cosmetic Applications

Mucuna pruriens is a medicinal plant whose seeds have various types of pharmacological activities and are used in many traditional medicines. The aim of this study was to evaluate the phytochemicals as well as the anti-aging, antioxidant, and moisturizing properties of seed extracts of M. pruriens var. pruriens and M. pruriens var. utilis. In addition, the best extract was selected for the development of nanoemulsions. M. pruriens var. utilis had the highest total phenolic and total flavonoid contents. It had good antioxidant activity (the IC50 of DPPH was 4.87 µg/mL, the FRAP value was 1.63 mg of FeSO4/mg of extract, and the percentage of lipid peroxidation was 80.19%) and anti-aging activity (the percentages of inhibition of hyaluronidase, collagenase, and elastase were 26.41%, 51.16%, and 22.78%, respectively). The occlusive factor was 46.12 ± 1.72. Therefore, M. pruriens var. utilis seed extract was selected for the preparation of nanoemulsions. The results showed that the size, PDI, and zeta potential of nanoemulsions containing M. pruriens var. utilis seed extract at day 30 did not significantly differ from those at day 0. In addition, the %EE was 63.46%. A study of skin permeation showed that the retention in the membrane after six hours of skin permeation study was 44.19%. Therefore, nanoemulsions containing M. pruriens var. utilis seed extract have good potential for further use in cosmetic applications.

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.

Sivelestat in Patients at a High Risk of Postoperative Acute Lung Injury After Scheduled Cardiac Surgery: A Prospective Cohort Study.

Sivelestat, a neutrophil elastase inhibitor, is specifically developed to mitigate the occurrence of acute lung injury (ALI) in individuals who are undergoing cardiovascular surgery. However, its impact on patients who are at a heightened risk of developing ALI after scheduled cardiac surgery has yet to be determined. In order to address this knowledge gap, we undertook a study to assess the efficacy of sivelestat in protecting the lungs of these patients. We conducted a prospective cohort study involving 718 patients who were at high risk of developing postoperative acute lung injury (ALI) and underwent scheduled cardiac surgery between April 25th, 2022, and September 7th, 2023. Among them, 52 patients received sivelestat (administered at a dosage of 0.2mg/kg/h for 3 days), while 666 patients served as controls, not receiving sivelestat. The control conditions were the same for all patients, including ventilation strategy, extubating time, and fluid management. Subsequently, a propensity-score matched cohort was established, consisting of 40 patients in both the sivelestat and control groups. The primary outcome measure encompassed a composite of adverse outcomes, including 30-day mortality, ALI, acute respiratory distress syndrome (ARDS), and others. Secondary outcomes assessed included pneumonia, ventricular arrhythmias, mechanical ventilation (MV) time, and more. After conducting propensity matching in our study, we observed that there were no significant differences in 30-day mortality between the sivelestat and control groups (0% vs 2.5%, P=0.32). However, the use of sivelestat exhibited a significant reduction in the incidence of acute lung injury/acute respiratory distress syndrome (ALI/ARDS) compared to the control group (0% vs 55%, P<0.01), pneumonia (0 vs 37.5%, P<0.01), MV time (median:8 hours, IQR:4-14.8 hours vs median: 15.2 hours, IQR:14-16.3 hours, P<0.01). Compared to the control group, the sivelestat could significantly decrease white cell count (P<0.01), neutrophile percentage (P<0.01) and C-reactive protein (P<0.01) in the period of postoperative 5 days. The prophylactic administration of sivelestat has shown promising results in reducing the occurrence of acute lung injury/acute respiratory distress syndrome (ALI/ARDS) in patients with a heightened risk of developing these conditions after elective cardiac surgery. Our study findings indicate that sivelestat may provide protective effects by suppressing inflammation triggered by neutrophil activation, thereby safeguarding pulmonary function. ChiCTR2200059102, https://www.chictr.org.cn/showproj.html?proj=166643.

Investigating Polypharmacology through Targeting Known Human Neutrophil Elastase Inhibitors to Proteinase 3.

Using a combination of multisite λ-dynamics (MSλD) together with in vitro IC50 assays, we evaluated the polypharmacological potential of a scaffold currently in clinical trials for inhibition of human neutrophil elastase (HNE), targeting cardiopulmonary disease, for efficacious inhibition of Proteinase 3 (PR3), a related neutrophil serine proteinase. The affinities we observe suggest that the dihydropyrimidinone scaffold can serve as a suitable starting point for the establishment of polypharmacologically targeting both enzymes and enhancing the potential for treatments addressing diseases like chronic obstructive pulmonary disease.

Sivelestat ameliorates sepsis-induced myocardial dysfunction by activating the PI3K/AKT/mTOR signaling pathway

The cardioprotective role of sivelestat, a neutrophil elastase inhibitor, has already been demonstrated, but the underlying molecular mechanism remains unclear. This study aimed to explore the mechanism underlying the role of sivelestat in sepsis-induced myocardial dysfunction (SIMD). We found that sivelestat treatment remarkably improved the viability and suppressed the apoptosis of lipopolysaccharide (LPS)-stimulated H9c2 cells. In vivo, sivelestat treatment was associated with an improved survival rate; reduced serum cTnT, TNF-α, IL-1β levels and myocardial TNF-α and IL-1β levels; ameliorated cardiac function and structure; and reduced cardiomyocyte apoptosis. Moreover, sivelestat treatment substantially increased Bcl-2 expression and suppressed caspase-3 and Bax expression in LPS-induced H9c2 cells and in the heart tissues of septic rats. Furthermore, the phosphatidylinositol 3-kinase/protein kinase B/mechanistic target of rapamycin (PI3K/AKT/mTOR) signaling pathway was activated both in vitro and in vivo. The protective effect of sivelestat against SIMD was reversed by the PI3K inhibitor LY294002. In summary, sivelestat can protect against SIMD by activating the PI3K/AKT/mTOR signaling pathway.

Clinical and psychopathological characteristics of patients with late-onset schizophrenia and schizophrenia-like psychoses in clusters identified by biological parameters

To assess clinical and psychopathological characteristics of late-aged female patients with late-onset psychoses in clusters formed on the basis of biochemical and immunological blood parameters. We examined 59 women with schizophrenia and schizophrenia-like psychoses with onset after 40 years (ICD-10 F20, F22.8, F25, F23, F06.2), including 34 women with late-onset (40-60 years) and 25 with very late onset psychoses (after 60 years). At the time of hospitalization, a clinical/ psychopathological study was carried out using CGI-S, PANSS, CDSS, and HAMD-17, as well as the activities of glutathione reductase (GR) and glutathione-S-transferase (GT) have been determined in erythrocyte hemolysates, and the activities of leukocyte elastase (LE) and α1-proteinase inhibitor (α1-PI) have been assessed in blood plasma. Biochemical and immunological parameters have been also determined in 34 age-matched mentally healthy women. Clustering by signs such as GR, GT, LE and α1-PI has yielded two clusters of objects (patients) significantly different in GT (p<0.0001), LE (p<0.0001), and α1-PI (p<0.001) activities. Relatively to the controls, in the cluster 1 patients, the activities of GST and α1-PI are increased, the activity of LE is decreased, whereas, in the cluster 2 patients, the activity of GR is decreased, and the activities of LE and α1-PI are increased. Cluster 1 patients differ from cluster 2 patients in greater severity of the condition (CGI-S, p=0.04) and higher total scores on PANSS subscales' items. Cluster 1 includes 76% of patients with very late onset. Different correlations between clinical and biological signs are found in two clusters. The identified clusters have different clinical and psychopathological characteristics. Dividing patients into subgroups according to biochemical and immunological parameters is promising for the search for differentiated therapeutic approaches.

Investigation of the effects of antimicrobial peptides and vitamin D in breast cancer

Breast cancer is the most common cancer in women. In recent years, there have been studies on the effectiveness of cathelicidins, one of the antimicrobial peptides, especially in breast cancer. Cathelicidin expression is regulated by a rather complex mechanism. Under normal conditions, vitamin D induces the release of cathelicidin from human neutrophils via Neutrophil Elastase (NE). Elafin, on the other hand, inhibits cathelicidin as a Neutrophil Elastase inhibitor. However, the effect of these molecules on tumor progression is unclear. In our study, we aimed to investigate the effects of vitamin D and Elafin on cathelicidin in breast cancer patients. Cathelicidin, Elafin, and Vitamin D Receptor levels in the pre and post-operative blood samples of control patients and patients diagnosed with breast cancer were analyzed by western blot method. According to our findings, it was observed that cathelicidin levels were especially high and elafin levels were low in the preoperative group. Since an inflammatory environment occurs in the presence of a tumor, there is an increase in vitamin D receptors and elafin in the postoperative group. It was thought that the low Elafin in the preoperative group caused vitamin D to secrete more Cathelicidin and increased the progression of the tumor, but the formation of cathelicidin was inhibited due to the increase in Elafin levels in the postoperative period. Thus, it was concluded that cathelicidin, which has an effect on the development of breast cancer, can be controlled by Elafin and Vitamin D receptors.

A Comprehensive Study on Chemical and Biological Investigation of Thymus Brachychilus Jalas: A Rich Source of Ursolic and Oleanolic Acids

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.

TF/PAR2 Signaling Axis Supports the Protumor Effect of Neutrophil Extracellular Traps (NETs) on Human Breast Cancer Cells.

Neutrophil extracellular traps (NETs) have been implicated in several hallmarks of cancer. Among the protumor effects, NETs promote epithelial-mesenchymal transition (EMT) in different cancer models. EMT has been linked to an enhanced expression of the clotting-initiating protein, tissue factor (TF), thus favoring the metastatic potential. TF may also exert protumor effects by facilitating the activation of protease-activated receptor 2 (PAR2). Herein, we evaluated whether NETs could induce TF expression in breast cancer cells and further promote procoagulant and intracellular signaling effects via the TF/PAR2 axis. T-47D and MCF7 cell lines were treated with isolated NETs, and samples were obtained for real-time PCR, flow cytometry, Western blotting, and plasma coagulation assays. In silico analyses were performed employing RNA-seq data from breast cancer patients deposited in The Cancer Genome Atlas (TCGA) database. A positive correlation was observed between neutrophil/NETs gene signatures and TF gene expression. Neutrophils/NETs gene signatures and PAR2 gene expression also showed a significant positive correlation in the bioinformatics model. In vitro analysis showed that treatment with NETs upregulated TF gene and protein expression in breast cancer cell lines. The inhibition of ERK/JNK reduced the TF gene expression induced by NETs. Remarkably, the pharmacological or genetic inhibition of the TF/PAR2 signaling axis attenuated the NETs-induced expression of several protumor genes. Also, treatment of NETs with a neutrophil elastase inhibitor reduced the expression of metastasis-related genes. Our results suggest that the TF/PAR2 signaling axis contributes to the pro-cancer effects of NETs in human breast cancer cells.

Promising Anti-Wrinkle Applications of Aromatic Extracts of Hedychium coronarium J. Koenig via Antioxidation and Collagenase Inhibition.

This study aimed to extract aromatic compounds from the rhizomes, leaf sheaths, and leaves of Hedychium coronarium and investigate their chemical compositions, cosmetic/cosmeceutical activities, and irritation potency. The chemical compositions were investigated via gas chromatography-mass spectrometry. The antioxidant activities were evaluated via spectrophotometry. The anti-skin wrinkle properties were investigated via collagenase, elastase, and hyaluronidase inhibition. The irritation potency was observed via a hen's egg-chorioallantoic membrane test. Eucalyptol was detected as a major component in the rhizomes and leaf sheaths, while β-caryophyllene was predominant in the leaves. The absolutes from the rhizomes were the strongest antioxidants, with ABTS scavenging properties similar to L-ascorbic acid. Interestingly, the equivalent concentration (EC1) of the absolute from the rhizome was 0.82 ± 0.01 µg FeSO4/g extract, which was significantly more potent than L-ascorbic acid (0.43 ± 0.03 µg FeSO4/g extract). The rhizome-derived absolute was the most effective against collagenase, while the concretes from the rhizomes and leaf sheaths showed promising anti-hyaluronidase activity with inhibitions of 90.5 ± 1.6% and 87.4 ± 5.1%, respectively. The irritability of the aromatic extracts was not different from that of the vehicle control, proving their safety. Therefore, the Hedychium coronarium rhizome-derived absolute was an attractive and potent antioxidant with anti-collagenase activities, indicating its potential for use in anti-aging formulations.