Search Results (290)

Additives such as surfactants (Tween-20) and cryoprotectants (glycerol and trehalose) are often used in enzymatic assays to improve the quality and long-term stabilization of proteins. However, these additives can affect the enzymatic activity and the enzyme’s affinity for active compounds, such as inhibitors, and must be considered during assay design since a slight shift in enzyme behavior may compromise the reliability of the results. In this study, the effects of Tween-20, glycerol, and trehalose on hyaluronidase (Hyal) were systematically evaluated by assessing their influence both directly—through microscale thermophoresis (MST) signals of the labeled enzyme (Hyal*)—and indirectly, by monitoring the formation of the final product of the degradation of hyaluronic acid, tetrasaccharide (Tet), using capillary electrophoresis (CE/UV). Hyal was labeled for the first time with ATTO-647 NHS ester, a commercial dye compatible with MST. Efficient labeling was achieved in a phosphate-based buffer without loss of catalytic activity. Tween-20 showed no impact on MST signals nor on enzymatic performance when used between 0.005 and 0.05% (v/v). Glycerol also did not interfere with MST measurements; however, it significantly reduced catalytic activity at concentrations above 2% (v/v). Trehalose affected Hyal* fluorescence in a concentration-dependent manner and enhanced catalytic activity even at 0.02% (v/v). Full article

Snakebite envenoming is a neglected tropical disease contributing to a significant number of morbidities and mortalities globally. Reports indicate that venom variation influences antivenom efficacy, which might affect treatment outcomes. The venom composition of Daboia russelii (Russell’s viper), one of the big four snakes in India, has been extensively studied from different geographical regions of India. Nonetheless, the Russell’s viper venom proteome from Kerala (Western Ghats region), together with its study in comparison with the same species’ venom from Tamil Nadu, has not been explored yet. In the current study, Daboia russelii venom from Irula (RVi) and the Western Ghats region in Kerala (RVwg) was characterized through mass spectrometry-based proteomics and few functional assays. The proteomics study identified 52 proteins from 14 snake protein families in RVi and 61 proteins from 17 snake venom protein families in RVwg. Some of the protein families, including DNase and hyaluronidase, as well as vascular endothelial growth factor, were exclusively identified in RVwg venom. Comparative functional analysis indicated that RVwg exhibited higher fibrinogenolytic and hyaluronidase activities, while RVi venom showed higher phospholipase A₂ and L-amino acid oxidase activities. Through ELISA, RVi venom showed an end-point titration value of 1:24,300 for all the antivenoms used in this study, whereas for RVwg, compared to PSAV (Premium serums and vaccines) (1:2700), Virchow and VINS (both 1:8100) antivenoms showed better immunological cross-reactivity. Immunoblotting experiments indicated differential binding and recognition of antigenic epitopes present in both venoms by the polyvalent antivenoms used in the current study. All these findings highlight that the venom proteome varies according to the geographical location, and this significantly influences antivenom efficacy. Full article

Background: Hyaluronic acid (HA) dermal fillers are among the most widely used injectable materials in esthetic medicine. They are generally safe, but delayed inflammatory reactions (DIRs) have been observed, particularly after viral infections or vaccinations. Such events have raised questions about the role of immune activation in filler-related complications. Objective: This review examined the available literature on DIRs to HA fillers that occurred in the context of viral illness or immunization, with attention to how these reactions present and how they are managed. Methods: A systematic search was carried out in PubMed, ScienceDirect, ClinicalKey, and Google Scholar between October and November 2024. Only human case reports and case series were included. The protocol was registered in PROSPERO (CRD420251030918), and study quality was assessed using the Newcastle–Ottawa Scale. Results: Six publications met inclusion criteria: four case series and two case reports, describing 25 women between 22 and 65 years of age. Patients developed swelling, erythema, angioedema, or, in severe cases, marked facial edema after HA filler injections, with symptom onset ranging from several hours to several weeks following viral exposure. Corticosteroids and hyaluronidase were the most common treatments, though milder cases sometimes resolved without intervention. Study quality varied, with some reports providing limited detail on patient characteristics and follow-up. Conclusions: DIRs associated with viral infections or vaccinations remain uncommon but clinically relevant complications of HA filler use. Limited case-based evidence indicates potential effectiveness of corticosteroids and hyaluronidase, though management practices remain inconsistent. Larger prospective studies are needed to clarify underlying mechanisms and to establish standardized guidelines for treatment. Full article

One of the main targets for snake venoms in animal and human organisms is the circulatory system. Mechanisms of circulatory system injury within the victim’s body include, among others, the direct effect of snake toxins on structures in blood vessel walls. The interaction of a toxin with cells and the extracellular matrix of the vessel wall may manifest as cytotoxicity, leading to cell death by necrosis or apoptosis, and damage to vascular wall structures. Such interactions may increase capillary permeability, promoting hemorrhage or edema, and may also induce alterations in vascular tone, resulting in changes in blood pressure. Snake toxins may also affect the growth, function, and regenerative ability of the endothelium, thus modulating angiogenesis; some toxins exert protective or anti-atherosclerotic effects. Toxins interacting with the vasculature may be classified as enzymes (phospholipases A₂, metalloproteinases, L-amino acid oxidases, and hyaluronidases), proteins without enzymatic activity (vascular endothelial growth factors, disintegrins, C-type lectins and snaclecs, three-finger toxins, etc.), peptides (bradykinin-potentiating peptides, natriuretic peptides, sarafotoxins), and low-molecular-weight substances. This review summarizes the data on the vascular effects, particularly on the blood vessel wall, exhibited by various classes and groups of snake toxins. Full article

In this study, the cosmeceutical potential of a 70% ethanol extract of Rosa lucieae was investigated as a multifunctional bioactive ingredient. The extract was systematically evaluated for its antioxidant, anti-melanogenic, and anti-aging properties, and was comprehensively phytochemically profiled using ultra-high-performance liquid chromatography–quadrupole time-of-flight mass spectrometry. The analysis tentatively identified 21 metabolites, including phenolic acids (gallic acid, ellagic acid, and corilagin), flavonoids (catechin, rutin, quercetin, hyperoside, and quercitrin), and glycosidic derivatives (e.g., phlorizin), several of which are well-documented for their skin-protective effects. Quantitative measurements confirmed high polyphenol and flavonoid contents, correlating with strong radical-scavenging and reducing capacities in α-diphenyl-β-picrylhydrazyl, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid, as well as ferric ion reducing antioxidant power assays. Moreover, the extract inhibited tyrosinase activity and 3,4-dihydroxyphenylalanine oxidation, thereby suppressing melanin biosynthesis. In addition, marked inhibitory effects against collagenase, elastase, and hyaluronidase were observed; these enzymes are critically involved in extracellular matrix degradation and skin aging. Taken together, these results indicate that the biological activities of R. lucieae are supported by a diverse polyphenol- and flavonoid-rich chemical profile, highlighting the potential of this plant as a natural multifunctional ingredient for cosmeceutical, nutraceutical, functional food, and preventive healthcare applications. Full article

Background/Objectives: The health benefits of Tamus communis fruits have been associated with their high phenolic content, which comprises several flavonoids. However, the extraction methods might significantly impact these valuable compounds’ bioactivity. Therefore, the current study assesses how different extraction techniques affect T. communis extracts’ antioxidant, anti-aging, antimicrobial, cytotoxic, anti-inflammatory, and phenolic contents. Methods: Conventional method (TCE-CM) and ultrasound-assisted extraction (TCE-UM) were the methods employed. Results: The increased phenolic content of TCE-UM, particularly flavonoids and phenolic acids, was demonstrated to be a contributing factor to its higher biological activity. Key enzymes linked to dermatological conditions, such as elastase, collagenase, hyaluronidase, and tyrosinase, were significantly inhibited by both extracts at 1 mg/mL; TCE-UM showed the highest tyrosinase inhibition (65.61  ±  5.21%) compared to TCE-CM (21.78  ±  2.19%). TCE-UM also demonstrated exceptional antibacterial performance, showing notable antibiofilm and metabolic inactivation effects and potent activity against pathogens such as Staphylococcus aureus, Escherichia coli, and Candida albicans. Both extracts showed concentration-dependent anti-inflammatory properties; TCE-UM had a lower IC50 value (26.46 ± 2.30%) in nitric oxide inhibition tests, suggesting stronger anti-inflammatory capabilities. Conclusions: These findings underscore the superior bioactivity of TCE-UM and suggest that ultrasonic extraction is a more efficient method for isolating bioactive phenolic compounds from T. communis fruits, presenting promising applications in anti-aging and antimicrobial formulations. Full article

The use of cyanobacteria has gained considerable interest in many industries, including the cosmetic industry, due to its rich array of bioactive metabolites. This study evaluates the in vitro photoprotective properties and the effect of Cylindrospermum alatosporum (NR125682) and Loriellopsis cavernicola (NR117881) extracts on slowing down the enzymes associated with skin aging. Various crude extracts were prepared using hexane, dichloromethane, and ethanol solvents. The resulting crude extract solvents were completely distilled to obtain their bioactive compounds, based on selected polarities. The sulfhydryl content of the crude extracts was determined and the aging-associated enzymes’ activity (collagenase, elastase, hyaluronidase, and tyrosinase) in the crude extracts was investigated. Furthermore, the in vitro photoprotective activity of the extracts was assessed by measuring UVA and UVB photoprotection. Most of the extracts contained varying amounts of sulfhydryl compounds (10.88–78.15 mg/g). All of the extracts demonstrated in vitro inhibitory activity against tyrosinase, hyaluronidase (IC₅₀ 6 µg/mL), and collagenase (IC₅₀ 50–70 µg/mL); weak elastase inhibitory activities were also observed. The crude extracts also showed appreciable UVA and UVB photoprotective activity. Meanwhile, L. cavernicola extracts demonstrated the highest UVB photoprotective activity (SPF 14.67–78.96). It is noteworthy that the crude extracts possessed anti-skin-aging potency with notable photoprotective capability. Full article

Hyaluronidase is a hydrolytic enzyme that cleaves β-1,4-glycosidic linkages in high-molecular-weight hyaluronic acid, generating low-molecular-weight oligosaccharides with enhanced biological functions. These products exhibit immunomodulatory, antioxidant, and tissue-regenerative properties, making them valuable in pharmaceutical, cosmetic, and functional food applications. However, most commercial hyaluronidases originate from pathogenic bacteria or recombinant hosts, raising concerns over their biosafety and regulatory acceptance, particularly in food-grade applications. In this study, we report the isolation and characterization of a novel non-pathogenic soil bacterium, Paenibacillus residui BSSK58, which produces an extracellular hyaluronidase. Whole-genome sequencing revealed the absence of known virulence factors and antibiotic resistance genes. Phenotypic safety evaluations confirmed that there was no hemolytic activity, biogenic amine production, or cytotoxicity against human intestinal epithelial cell lines (Caco-2 and HT-29). The purified BSSK58 hyaluronidase exhibited a molecular weight of approximately 170 kDa, with optimal activity at pH 8.0–9.0 and 50 °C. The enzyme showed broad substrate specificity toward hyaluronic acid, chondroitin sulfate, and alginate, and its depolymerizing activity was confirmed using gel permeation chromatography. Furthermore, a 13-week oral repeated-dose toxicity study under Good Laboratory Practice conditions demonstrated no adverse effects. These findings support the use of BSSK58 hyaluronidase as a safe, non-recombinant biocatalyst suitable for industrial applications under regulatory-compliant frameworks. Full article

Hamamelis virginiana (witch hazel) is traditionally used in dermatology for its antibacterial and anti-inflammatory effects. However, the number of studies on its chemical composition and potentials in skin protection remains limited. This study aimed to investigate the qualitative and quantitative composition of polyphenolic compounds in the leaves and bark of the plant, as well as to explore their antioxidant, anti-inflammatory, and extracellular matrix (ECM)-protective activities in skin-relevant cell models. Human dermal fibroblasts and keratinocytes were exposed to oxidative and inflammatory stimuli and pretreated with leaf and bark extracts. ROS levels, antioxidant enzyme activity (SOD, GPx, CAT), pro-inflammatory cytokines (IL-6, IL-1β, TNF-α), and inhibition of collagenase, hyaluronidase, and elastase were assessed. Both extracts strongly reduced ROS levels, enhanced SOD activity, and significantly decreased pro-inflammatory cytokines. Bark extract also exhibited potent inhibitory activity against collagenase and elastase. UPLC-DAD-MS analysis revealed that both plant parts contained high levels of tannins; however, the leaf extract showed a more diverse composition, including more complex tannin forms and a significant amount of flavonoids from the quercetin and kaempferol class. In conclusion, H. virginiana leaf and bark extracts demonstrate multifunctional antioxidant and anti-inflammatory properties, supporting their potential use in cosmeceuticals and dermatological formulations targeting skin aging and inflammation. Full article

The hyaluronidase enzyme derived from Vespa tropica (VesT2a) venom contains two putative catalytic residues. Herein, a double mutation was introduced into VesT2a at its catalytic sites by substituting Asp107 and Glu109 with Asn and Gln, respectively, to assess their essential roles in enzymatic function. We used Pichia pastoris to produce the mutated version of the VesT2a (mVesT2a) protein, and the process was more efficient when employing the methanol-inducible promoter (P_AOX1) compared to the constitutive promoter (P_GAP). In bioreactor scale-up, P. pastoris harboring the pAOX1-αMF-mVesT2a plasmid secreted 34.03 ± 2.31 mg/L of mVesT2a, with an apparent molecular mass of 46.6 kDa, retaining only 2.9% of hyaluronidase activity, thus indicating successful mutation. The newly developed indirect ELISA-based method using mVesT2a demonstrated its potential as an alternative approach for measuring hyaluronic acid (HA) at low concentrations and was also used to confirm HA-binding capacity. In silico docking and molecular dynamics simulations further supported the stable interaction of the mVesT2a–HA complex while suggested other surrounded acidic amino acid residues, which may play a minor role in HA degradation, supporting the remaining activity observed in the in vitro experiments. Full article

Opuntia stricta var. dillenii (OPD) fruits are rich in betalains and phenolic compounds, which are recognized for their potential health-promoting properties. This study focuses on the optimization of pulsed electric field (PEF)-assisted solid–liquid green extraction (SLE) from OPD whole fruit, using response surface methodology (RSM) experimental design to obtain green extracts rich in bioactive compounds. The optimal PEF pre-treatment conditions (electric field strength and energy input) were determined based on the cell disintegration index (Zp), followed by optimizing SLE conditions (temperature, time, and ethanol content). High-performance liquid chromatography (HPLC-DAD-ESI-Qtof) was used to characterize the individual bioactive compound profile of the obtained OPD green extracts. Results showed that optimal PEF pre-treatment conditions were at 10.5 kJ/kg and 5 kV/cm, followed by SLE at 35 °C for 165 min, using water as the solvent. Conventional optimal SLE conducted at 45 °C, 8% ethanol, and 128 min was applied as the control process. The combined PEF-assisted SLE process enhanced total betalain and phenolic compound yields by 61% and 135%, respectively. Antioxidant activities (DPPH by 145%, FRAP by 28%) and anti-inflammatory potential (hyaluronidase inhibition by 19%) were also significantly improved. This study underscores the potential use of a PEF pre-treatment to improve obtaining green extracts rich in bioactive compounds with high biological activities from OPD whole fruits, using water as a solvent. Full article

Type 2 diabetes is a serious public health problem in the 21st century. To find new substances supporting diabetes therapy, researchers are increasingly paying attention to the biological potential of edible flowers. This study assessed the antidiabetic potential of ethanol, 50% ethanol, and water extracts from Paeonia officinalis L., Forsythia × intermedia, Gomphrena globosa L., and Clitoria ternatea L. flowers. Extracts were tested for antioxidant activity (DPPH, ABTS, FRAP, CUPRAC, and Fe²⁺ chelation), enzyme inhibition (α-glucosidase, α-amylase, hyaluronidase, and cholinesterases), and anti-inflammatory effects (NO inhibition in LPS-stimulated RAW264.7 macrophages). Phytochemical composition was also analysed. Extracts of P. officinalis stood out with the highest total phenolic content (50% ethanol extract of P. officinalis 178.49 mg GAE/g) and total flavonoid content (aqueous extracts of P. officinalis 4.27 mg QE/g), high gallic acid level, and the effective inhibition of α-glucosidase and α-amylase (α-glucosidase inhibition 98–99% for all P. officinalis extracts, and α-amylase inhibition ~ 100% for ethanolic extract). Strong hyaluronidase (76.9–95.5%) and cholinesterase inhibition was also observed. F. × intermedia extracts were rich in rutin and chlorogenic acid and showed potent inhibitory effects on α-glucosidase (50% ethanol extract 91.59%), α-amylase (aqueous extract 89.35%), and hyaluronidase (aqueous extract 73.8%). Ethanol extracts of G. globosa exhibited a high α-amylase inhibition (93–95%). Although C. ternatea showed moderate antioxidant activity, it showed an apparent anti-inflammatory effect, effectively reducing NO production in activated macrophages for 50% ethanol extract. In summary, P. officinalis and F. × intermedia flowers are promising sources of extracts with antioxidant, antidiabetic, and anti-inflammatory effects supporting their use in further research on type 2 diabetes therapy. Full article

Equisetum telmateia Ehrh. (great horsetail) belongs to the Equisetaceae family and its aerial parts have been traditionally used for skin conditions and to achieve healthy and resilient skin, nails, and hair. This study aimed to evaluate the inhibition of skin-related enzymes by, the antioxidant capacity of, and the phytochemical composition of E. telmateia. Additionally, a novel emulgel was formulated from the main methanolic extract and characterized in terms of pH, viscosity, determination of content quantification, textural profile analysis, and spreadability. After the characterization studies, in vitro release and ex vivo permeation and penetration studies were performed. Firstly, the dried aerial parts of E. telmateia were macerated in methanol, followed by partitioning with solvents of increasing polarity: n-hexane, chloroform, ethyl acetate, and n-butanol. Antioxidant activity was assessed using DPPH, FRAP, CUPRAC, and TOAC assays, while enzyme inhibition was analyzed for collagenase, elastase, hyaluronidase, and tyrosinase. LC-MS/MS analysis identified 53 phytochemical compounds. Protocatechuic acid, the main phenolic compound, was quantitatively analyzed in each subfraction by HPTLC. The in vitro release studies showed sustained release of the reference substance (protocatechuic acid) and the kinetic modeling of the release was fitted to the Higuchi model. The ex vivo permeation and penetration studies showed that the formulation exhibited a retention of 3.06 ± 0.21 µg.cm⁻² after 24 h, whereas the suspended extract demonstrated a skin retention of 1.28 ± 0.47 µg.cm⁻². Both the extracts and the formulated emulgel exhibited inhibitory effects on skin-related enzymes. Our finding suggested that E. telmateia might be a valuable ingredient for wrinkle care and skin-regenerating cosmetics. Full article

Carnivorous plants have garnered attention as sources of pharmacologically active compounds, yet their floral tissues remain largely underexplored. In this study, we investigated the bioactive properties of Nepenthes miranda flower extracts prepared using water, methanol, ethanol, and acetone. Among these, the ethanol extract exhibited the highest total phenolic content (18.2 mg GAE/g), flavonoid content (68.9 mg QUE/g), and antioxidant activity (DPPH IC₅₀ = 66.9 μg/mL), along with strong antibacterial effects against Escherichia coli and Staphylococcus aureus. Cosmetically relevant enzyme inhibition assays revealed significant activity against tyrosinase (IC₅₀ = 48.58 μg/mL), elastase (IC₅₀ = 1.77 μg/mL), and hyaluronidase (IC₅₀ = 7.33 μg/mL), supporting its potential as an anti-skin aging agent. For antidiabetic evaluation, the ethanol extract demonstrated potent α-glucosidase inhibition (IC₅₀ = 24.53 μg/mL), outperforming standard inhibitors such as acarbose and quercetin. The extract also displayed marked cytotoxicity against A431 epidermoid carcinoma cells (IC₅₀ = 90.61 μg/mL), inducing dose-dependent apoptosis, inhibiting cell migration and colony formation, and causing significant DNA damage as shown by comet assay. Furthermore, the ethanol extract strongly inhibited the activity of purified human dihydroorotase (IC₅₀ = 25.11 μg/mL), indicating that disruption of pyrimidine biosynthesis may underlie its anticancer activity. Overall, this study provides the first characterization of N. miranda flower extracts, particularly the ethanol fraction, as a promising source of multifunctional bioactive compounds with possible applications in cosmetics, antidiabetic therapy, and cancer treatment. Full article

Scorpionism is a growing public health concern in Brazil, with the Amazon region presenting the highest mortality rates but remaining understudied, especially regarding local scorpion venoms composition. This study presents the first comprehensive biochemical characterization of venoms from three Amazonian species—Tityus metuendus (TmetuV), Tityus silvestris (TsilvV), and Brotheas amazonicus (BamazV)—using an integrated approach combining Multi-Enzymatic Limited Digestion (MELD)-based bottom-up proteomics, high-resolution LC-MS/MS, chromatography, zymography, and enzymatic assays. Tityus serrulatus venom was included as a reference. Significant biochemical differences were observed: TsilvV was rich in 20–30 kDa proteins and showed strong metalloprotease activity; BamazV exhibited high molecular weight proteins and potent phospholipase A₂ (PLA₂) activity but lacked proteolytic and fibrinogenolytic activities; TmetuV showed the highest hyaluronidase activity and abundance of α-KTx neurotoxins. Zymography revealed a conserved ~45 kDa hyaluronidase in all species. Three novel components were partially characterized: BamazPLA₂ (Group III PLA₂), Tmetu1 (37-residue α-KTx), and TsilvMP_A (a metalloprotease homologous to antarease). This is the first application of MELD-based proteomics to Amazonian scorpion venoms, revealing molecular diversity and functional divergence within Tityus and Brotheas, emphasizing the need for region-specific antivenoms. These findings provide a foundation for future pharmacological studies and the discovery of bioactive peptides with therapeutic potential. Full article