Search Results (3,384)

Background/Objectives: Plants and fruits of Physalis philadelphica Lam. Solanacea are commonly used in traditional medicine to improve some illnesses such as diabetes, in North and Central American countries. The aim was to evaluate the effects of aqueous maceration (He-M) and ultrasound-assisted (He-US) extracts of P. philadelphica husk on hyperglycemia, insulin resistance, hepatic steatosis, and oxidative stress in obese rats. Methods: The effects of husk extracts on carbohydrate and lipid absorption were evaluated using oral starch and lipid tolerance tests in healthy male Wistar rats. Obesity was then induced using a high-fructose and saturated fat diet, followed by 16 weeks of extract administration. Results: He-US significantly reduced the postprandial glycemic spike, while both extracts lowered serum triglyceride levels (~50%) following lipid loading, compared with the negative control. In obese rats, both extracts reduced body weight gain (~10%) and lowered fasting glucose levels (22% for He-M and 15% for He-US), compared with the obese control. He-US also reduced insulin levels (~32%), insulin resistance (~53%), and free fatty acids (~52%), while He-M improved hepatic steatosis and reduced liver triglycerides (~26%). Both extracts reduced hepatic nitrite levels, although only He-M significantly decreased lipid peroxidation (~32%). Additionally, both treatments enhanced hepatic antioxidant enzyme activity. Conclusions: Husk extracts exerted beneficial effects on hyperglycemia, insulin resistance, hepatic steatosis, and oxidative stress markers in obese rats. Full article

Compost amendments are widely recognized as an effective strategy for improving soil quality, modulating enzyme activities, and enhancing nitrogen cycling. Urease, a key enzyme in nitrogen transformation, is characterized by kinetic parameters such as the maximum reaction rate (V_max) and Michaelis constant (K_m), as well as thermodynamic attributes including temperature sensitivity (Q₁₀), activation energy (E_a), enthalpy change (ΔH), Gibbs free energy change (ΔG), and entropy change (ΔS). However, how different compost sources regulate urease kinetics, thermodynamics, and nitrogen availability remains poorly understood. In this study, we evaluated the effects of three compost amendments—mushroom residue (MR), mushroom residue–straw mixture (MSM), and leaf litter (LL)—on urease kinetics and thermodynamics in a temperate agroecosystem. The MSM treatment significantly enhanced urea hydrolysis capacity and catalytic efficiency. In contrast, LL treatment resulted in the highest Km value, indicating a substantially lower enzyme-substrate affinity. Furthermore, MSM reduced the E_a and increased the thermal stability of urease, thereby supporting enzymatic performance under fluctuating temperatures. Collectively, our findings highlight that compost composition is a critical determinant of urease function and nitrogen turnover. By elucidating the coupled kinetic and thermodynamic responses of urease to compost inputs, this study provides mechanistic insights to guide optimized soil management and sustainable nitrogen utilization in temperate agricultural systems. Full article

In this study, we constructed an extraction process for bovine lung peptide-1 (BLP-1) derived from bovine lung tissue utilizing single-factor optimization combined with response surface methodology. We systematically analyzed its antioxidant activity, biological safety, and therapeutic mechanisms against alcoholic liver disease (ALD). In vitro experiments demonstrated that BLP-1 exhibits excellent scavenging activity against various free radicals, while exhibiting no significant cytotoxicity or hemolytic activity. In a model of H₂O₂-induced oxidative damage in HepG2 cells, BLP-1 significantly alleviated oxidative stress injury by upregulating the activities of intracellular antioxidant enzymes. Animal experiments further confirmed that BLP-1 significantly reduced serum levels of transaminase, inhibited the release of inflammatory factors, enhanced antioxidant enzyme activity, and ameliorated lipid peroxidation and pathological injury in ALD mice. By combining liquid chromatography-tandem mass spectrometry (LC-MS/MS) with bioinformatics, we screened 12 novel antioxidant peptides. Among these, the binding energies of GP9, FG6, and WG6 to Keap1 were −10.2, −9.7, and −8.7 kcal/mol, respectively, indicating their potential to modulate the antioxidant defense system through competitive inhibition of Keap1-Nrf2 interactions. This study provides a novel approach for the high-value utilization of bovine lung and the treatment of ALD, as well as a new source for the extraction of natural antioxidant peptides. Full article

Lipases (EC 3.1.1.3) catalyze the hydrolysis of triacylglycerols into mono- and diacylglycerols and free fatty acids. This study investigated the production of lipase by Hypocrea pseudokoningii under solid-state fermentation (SSF), followed by its immobilization, purification, and biochemical characterization. Maximum activity was achieved using wheat fiber after 168 h of cultivation. Supplementation with oils enhanced production, particularly palm oil (315U; 1.58-fold increase) and soybean oil (Glycine max) (298U; 1.49-fold increase). The addition of micronutrients further improved yields, with Khanna (445U) and Vogel (400U) salts promoting more than a two-fold increase in activity. Immobilization on Octyl-Sepharose significantly altered the enzyme’s properties. The free lipase exhibited optimal activity at 45 °C and pH 4.5–5.5, while the immobilized enzyme showed maximum activity at 35–40 °C and pH 5.5. Thermal stability was notable enhanced: the free lipase had a half-life of 10 min at 50 °C, whereas the immobilized enzyme remained stable for 60 min and retained over 30% activity at 70 °C. Both the free and immobilized forms were stable across a broad pH range (4.0–10.0), maintaining more than 70% residual activity. The enzyme was stabilized by Tween 80 but inhibited by SDS. It was activated by Ca²⁺ and showed resistance to Pb²⁺, Zn²⁺, and Cu²⁺. Hydrolytic assays revealed murumuru (Astrocaryum murumuru), cupuaçu (Theobroma grandiflorum), and soybean oils as preferred substrates. TLC confirmed the formation of mono- and diglycerides, as well as the presence of fatty acids. Full article

In line with the circular economy approach and the pursuit of sustainable solutions for spent coffee grounds, this study investigates the valorization of spent coffee grounds as a source of cellulose-based enzyme immobilization carriers. Considering that global coffee consumption generates approximately 6.9 million tonnes of spent coffee grounds annually, their disposal represents both an environmental challenge and an opportunity for value-added applications. A multistep extraction process, including Soxhlet extraction followed by sequential subcritical extraction with ethanol and water, and alkaline treatment, led to the production of cellulose-enriched carriers. The carriers obtained were characterized by their morphology, porosity and surface properties and subsequently used for the two lipases immobilization, Burkholderia cepacia (BCL) and Pseudomonas fluorescens (PFL), using three techniques: adsorption and covalent binding via direct and indirect methods. The immobilized lipases were analyzed for key biochemical and operational properties and compared with each other and with their free enzymes. Based on their stability, catalytic activity, and reusability, the lipases immobilized by adsorption were identified as the most efficient biocatalysts. These immobilized enzymes were then used in two selected reactions to demonstrate their practical utility: cocoa butter substitute synthesis using PFL and the enzymatic pretreatment of wastewater from the oil processing industry using BCL. Both immobilized lipases showed excellent catalytic performance and maintained their high activity over four consecutive reuse cycles. Full article

Young leaves and sprouts of Toona sinensis are widely consumed throughout Asia. However, they are highly perishable, resulting in a short shelf life and limited marketability. Low-temperature storage can extend shelf life but is energy-intensive, restricting large-scale use. Salicylic acid (SA) exhibits preservative properties, representing a promising alternative. In this study, we investigated the effects of 250 μmol SA treatment on postharvest T. sinensis buds and leaves stored at 20 °C for 7 days and compared it to low-temperature storage (4 °C). SA treatment delayed wilting, reduced water loss, suppressed total soluble solid accumulation, and inhibited active oxygen free radical and malondialdehyde production compared to the untreated controls at room temperature while outperforming low-temperature storage. The treated buds also maintained higher antioxidant enzyme activity and preserved non-enzymatic antioxidant compounds, including ascorbic acid, polyphenols, and flavonoids. Although total amino acid content steadily increased under SA treatment, individual amino acids greatly fluctuated. SA treatment reduced nitrate reductase and glutamate dehydrogenase activities, as well as NO₂•⁻ levels, indicating its effects on nitrogen metabolism. Our findings indicate that SA improves the postharvest quality of T. sinensis buds during room-temperature storage, providing a theoretical basis for applying SA as a storage agent for T. sinensis buds. Full article

Melatonin, traditionally recognized for its role in regulating circadian rhythms and sleep, has emerged as a multifunctional molecule with significant implications in dermatology. Melatonin is described here as a pleiotropic, context-dependent modulator with antioxidant-related and immunomodulatory actions that are supported by both direct chemical scavenging in cell-free systems and indirect, enzyme-mediated effects in cells and tissues; its antitumor activity in dermatology is primarily preclinical and remains to be confirmed in large clinical trials. Melatonin protects skin cells from oxidative stress, UV radiation, and environmental damage by directly scavenging free radicals and activating endogenous defense systems. It also modulates immune responses, making it relevant in inflammatory dermatoses such as atopic dermatitis, while promoting tissue repair in wound healing and reducing signs of skin aging. Particular attention is given to topical formulations, including gels, creams, and patches, which enable localized delivery, improved skin penetration, and reduced systemic side effects. The review also discusses ongoing clinical trials, delivery technologies, and the potential for combinatorial therapies with established dermatological agents. Full article

Objectives: Meniscal integrity is crucial for knee joint stability and the prevention of osteoarthritis (OA) development. Recent studies suggested that mechanical overload and interleukin (IL)-17A may be important intertwined players in meniscal degeneration, but a direct impact of IL-17A on the meniscus has not been investigated. Therefore, the aim of this study was to analyze the effect of IL-17A on meniscal tissue with and without combined mechanical injury (MI). Methods: Meniscal explant disks (1 mm height, 3 mm diameter) were isolated from bovine menisci (preserving the native tibial superficial zone) and exposed to IL-17A [0–100 ng/mL] and/or MI (single compression, 50% strain, strain rate 1 mm/sec). After three days of incubation in a serum-free medium, the proteoglycan release (sGAG; DMMB assay), mRNA level of matrix-degrading enzymes (qRT-PCR), aggrecan degradation (NITEGE immunostaining), and cell death (histomorphometry of nuclear blebbing/apoptosis and condensed nuclei/unspecified cell death) were determined. Statistics: one- and two-way ANOVA with Tukey’s multiple comparisons or Kruskal–Wallis with post hoc testing. Results: IL-17A increased sGAG release in a dose-dependent significant manner. MI also induced the release of sGAG significantly, but the combination with IL-17A showed the highest levels. Both IL-17A and MI individually affected the mRNA levels for ADAMTS4 and MMP-13 slightly, but the combination of both particularly induced a significant increase in mRNA levels. Signals for the ADAMTS4-related aggrecan neoepitope NITEGE were elevated by IL-17A in superficial areas of the excised tissue and by MI in superficial and deeper areas. The combination of both stimuli intensified this signal further. MI increased the number of cells with condensed nuclei significantly and induced apoptosis in a small proportion of cells. IL-17A had no significant impact on the amount of condensed or apoptotic nuclei. Conclusions: Our findings emphasize an interaction between inflammatory cytokine IL-17A signaling and mechanical stress since IL-17A induced matrix degeneration in meniscal tissue, which intensified in combination with a trauma. The latter might create a post-traumatic environment that promotes meniscal degeneration and subsequently osteoarthritis progression. Full article

Phosphatidylserine (PS) holds considerable importance in both the food and medical sectors; however, its biosynthesis is critically dependent on phospholipase D (PLD). The practical application of PLD is constrained by pronounced side reactions in its free form and by reduced selectivity when immobilized. To address these challenges, this study employed a sequential strategy involving bioimprinting to hyperactivate PLD, followed by microencapsulation via ionotropic gelation within an alginate–chitosan matrix. This approach induced conformational rigidification, enabling PLD to maintain its hyperactivated state in aqueous environments. Under optimal conditions, the encapsulation efficiency reached 78.56%, and the enzyme activity recovery achieved 105.27%. The immobilized bioimprinted PLD demonstrated exceptional catalytic performance, achieving a 94.68% PS yield within 20 min, which significantly surpassed that of free PLD (85.82% in 150 min) and non-imprinted immobilized PLD (90.34% in 60 min). This represents 7.27-fold and 2.14-fold efficiency improvements, respectively. Furthermore, the biocatalyst exhibited outstanding storage stability, thermal stability, and reusability (77.53% yield after 8 cycles). To our knowledge, this is the first report combining bioimprinting with alginate-chitosan microencapsulation via ionotropic gelation, which yielded remarkably enhanced PLD activity. These findings highlight the strong potential of this method for efficient PS production. Full article

Shade-induced flower abscission in soybean plants is a significant factor limiting yield improvement. Under shaded conditions, significant differences exist in the flower abscission rates among different soybean varieties, but the regulatory mechanisms remain unclear. This study selected Tiedou 44 (T44) and Liaodou 32 (L32) as experimental materials. Results indicate that under shaded conditions, the flower abscission rate of T44 was significantly higher than that of L32. Physiological analysis revealed that cell wall degradation enzyme activity in T44 pedicels was significantly higher than in L32. Furthermore, compared to L32, T44 flowers under shade conditions exhibited significantly higher levels of IAA, IAA–amino acid conjugates, and ABA. The expression levels of PIN family genes (GMPIN3C, GMPIN3D, PIN3A, GMPIN1A, GMPIN1B, GMPIN1C, GMPIN1D, and GMPIN1E) in T44 were downregulated. These results suggest that the obstruction of auxin polar transport leads to auxin accumulation in flowers. This accumulation, in turn, triggers flower abscission. Additionally, GH3 gene expression was upregulated in T44 compared to L32. GH3 proteins catalyze the conjugation of free auxin (IAA) with amino acids, forming inactive IAA–amino acid complexes. This significantly reduces the concentration of free IAA capable of inhibiting abscission in T44, making flowers more prone to abscission. This study provides crucial insights into the molecular regulatory mechanisms underlying flower abscission in soybean. Full article

An intracellular α-glucosidase was isolated and purified from a Pseudanabaena sp. cyanobacterial strain. Before the enzyme purification, the optimal cultural conditions were determined. Optimal culture conditions (15 g/L maltose, 2 g/L yeast extract, 23 ± 1 °C) yielded 3.3 g/L of biomass and 2186 U/L of α-glucosidase in a lab-scale bioreactor. The purified enzyme displayed a molecular mass of 52 kDa with optimum activity at 40 °C and pH 7.0, and maintained stability within an acidic and neutral range of pH 4.0 to 7.0. Enzyme activity was affected by both the concentration and interaction time of the metal ions and chelator. Kinetic constants of K_m, V_max, and k_cat for the hydrolysis of pNPG were determined as 2.0 Mm, 2.9 μmol min⁻¹, and 14.86 min⁻¹, respectively. The activation energy (E_a) was 24.2 kJ mol⁻¹ and the thermodynamic parameters of enthalpy (ΔH^*), entropy (ΔS^*) of activation, Gibbs free energy (ΔG^*), free energy of substrate binding (ΔG^*_E-S), and transition state formation (ΔG^*_Ε-Τ) were 21.6, −116, 57.8, −22.2, and −41.2 kJ mol⁻¹, respectively. Moreover, the thermodynamic parameters for thermal inactivation of the enzyme were ΔH^*= 131 kJ mol⁻¹, 105 ≤ ΔS^* ≤ 108 kJ mol⁻¹, and 96 ≤ ΔG^* ≤ 98 kJ mol⁻¹, while the thermal inactivation energy (E_(a)d) was determined to be 133 kJ mol⁻¹. This is the first detailed investigation concerning the characterization of α-glucosidase derived from cyanobacteria. The presented enzymatic characteristics provide a valuable predictive model for identifying suitable applications. Full article

The enzymatic production of fatty acid ethyl esters (FAEEs) and xylose fatty acid esters (XFAEs) from soybean oil deodorizer distillate (SODD) was investigated using a hydroesterification strategy. SODD was enzymatically hydrolyzed, and the glycerol-free fraction was esterified with either xylose or ethanol. Free lipase from Pseudomonas fluorescens (PFL) yielded 84 wt% of free fatty acids (FFAs) production (with approximately 15% FFAs remaining as glycerides) after 48 h, using a SODD-to-water mass ratio of 1:4 and an enzyme loading of 5 wt% (based on oil mass). In the synthesis of FAEEs, free Eversa Transform converted approximately 82% of the FFAs into FAEEs after 48 h, using an ethanol-to-FFA molar ratio of 3.64:1 and an enzyme loading of 8.36% (w/v). For the synthesis of XFAEs, commercially immobilized lipases from Thermomyces lanuginosus (TLL-T2-150) and Pseudomonas fluorescens (IMMAPF-T2-150) were employed. These commercial lipase preparations are available in their immobilized form on Immobead T2-150. TLL-T2-150 resulted in a lower degree of xylose ester formation (80.20%) compared to IMMAPF-T2-150 (89.20%) after 24 h, using an FFA-to-xylose molar ratio of 5 in ethyl-methyl-ketone (xylose concentration of 7 mmol L⁻¹) and an enzyme loading of 0.5% (w/v). However, TLL-T2-150 consumed more FFAs, suggesting a higher degree of xylose esterification. The final reaction mixture containing XFAEs obtained with this biocatalyst exhibited suitable emulsifying properties. A Life Cycle Assessment (LCA) revealed that the enzymatic hydroesterification process offers a sustainable route for FAEEs and XFAEs production, with configurations using free PFL in hydrolysis and IMMAPF-T2-150 in esterification showing the lowest environmental impacts due to higher catalytic efficiency. The findings point to a clear environmental edge in using SODD for ester production, offering a cleaner alternative to standard methods and making better use of a renewable resource. Full article

This study explored the potential therapeutic effect and possible mechanism of Atractylodes macrocephala Koidz. Polysaccharide (AP) on pirarubicin chemotherapy-induced depression (CID) in breast cancer mice. This study utilized a variety of techniques to explore the potential of AP in mitigating behavioral abnormalities and elucidate the role of gut microbiota regulation in its therapeutic effects on chemotherapy in breast cancer mice. These included a chemotherapy mouse model, behavioral assessments, histological analysis using hematoxylin and eosin staining, ultrastructural examination, enzyme-linked immunosorbent assays, 16S rDNA sequencing, metabolomic profiling, Western blot analysis, and a pseudo-germ-free animal model. Oral administration of AP significantly improved depression-like behaviors in breast cancer chemotherapy mice while also reducing neuronal damage and inflammation in the hippocampus. AP prevented ferroptosis of intestinal tissues caused by chemotherapy and had a repairing effect on the intestinal barrier damage of chemotherapy-induced mice. Additionally, AP enhanced gut microbiota composition and altered intestinal metabolites in chemotherapy-treated mice. It notably decreased the abundance of certain microbes, such as Bacteroidaceae, Lachnospiraceae, Oscillospiraceae, and Clostridium, while significantly increasing the abundance of Alistipes. Moreover, AP efficiently modulated intestinal metabolites, including glycocholic acid, L-Phenylalanine, and palmitoylcarnitine. More importantly, depletion of gut microbiota through antibiotics diminished the effectiveness of AP. Our results suggest that AP alleviates depression-like behaviors in chemotherapy-treated mice by regulating the gut microbiota and microbial metabolism, as well as suppressing ferroptosis in intestinal tissues. Full article

Background/Objectives: Urinary angiotensin-converting enzyme (uACE) activity has long been regarded as a promising biomarker for kidney and cardiovascular diseases; however, its clinical applicability has been limited by the presence of endogenous urinary inhibitors and technically demanding assay protocols. We aimed to establish a fast and reproducible method for measuring uACE activity to identify the inhibitory compounds responsible for previous assay failures and to define practical preanalytical conditions suitable for routine laboratory implementation. Methods: A fluorescence-based kinetic assay was optimized for urine samples. Endogenous inhibitors were isolated by membrane filtration and chemically characterized, while the effect of sample dilution was evaluated as a simplified alternative for eliminating inhibitory interference. We assessed the stability of ACE activity under various storage conditions to support reliable measurement. Results: Urea (IC₅₀ = 1.18 M), uric acid (IC₅₀ = 3.61 × 10⁻³ M), and urobilinogen (IC₅₀ = 2.98 × 10⁻⁴ M) were identified as the principal reversible inhibitors, jointly accounting for up to 90% suppression of uACE activity. Their inhibitory effect was effectively eliminated by a 128-fold dilution. ACE activity remained stable for 24 h at 25 °C but was completely lost after freezing. A strong positive correlation between uACE activity and creatinine concentration (r = 0.76, p < 0.0001) justified normalization. ACE activity-to-creatinine ratio turned out to be significantly lower in ACE inhibitor-treated patients than in untreated controls (6.49 vs. 36.69 U/mol, p < 0.0001). Conclusions: Our findings demonstrate that accurate measurement of uACE activity is feasible using a rapid dilution-based protocol. The normalized ACE activity can serve as a practical biomarker for detecting pharmacological ACE inhibition and monitoring therapy adherence in cardiovascular care and may also provide insight into renal pathophysiology such as tubular injury or local RAAS-related processes. Full article

In the present study, seven previously undescribed resin glycosides, designated cusponins I-VII (1–7), together with one known analog (8), were isolated from the seeds of Cuscuta japonica, a traditional medicine used in China. Structural elucidation revealed them to be glycosidic acid methyl esters, generated through on-column methyl esterification of naturally occurring resin glycosides catalyzed by NH₂-functionalized silica gel. All isolates were characterized as either pentasaccharides or tetrasaccharides, incorporating D-glucose, L-rhamnose, or D-fucose units as the sugar residues. Notably, compounds 1 and 3–7 contained the uncommon aglycone, 11S-hydroxypentadecanoic acid. Bioactivity assessments demonstrated that compounds 1–4, 6 and 8 suppressed α-glucosidase activity, with IC₅₀ values between 8.02 and 71.39 μM. In addition, compounds 3 and 5 exhibited inhibitory effects on protein tyrosine phosphatase 1B (PTP1B), with IC₅₀ values of 14.19 ± 1.29 μM and 62.31 ± 8.61 μM, respectively, marking the first report of PTP1B inhibitory activity among resin glycosides. Enzyme kinetic analyses indicated that compound 2 acted as an uncompetitive α-glucosidase inhibitor (K_is = 3.02 μM), whereas compound 3 inhibited PTP1B via a mixed-type mechanism (Kᵢ = 24.82 μM; K_is = 64.24 μM). Molecular docking combined with molecular dynamics simulations suggested that compounds 2 and 3 interacted with α-glucosidase-pNPG and PTP1B, respectively, forming stable complexes with favorable binding free energies. Collectively, this study reported eight resin glycosides from C. japonica, seven of them newly identified, with compounds 2 and 3 highlighted as promising scaffolds for antidiabetic drug discovery. Full article