Search Results (1,373)

Background: Inflammation abrogates cellular organization and tissue homoeostasis, resulting in redness, swelling, heat, pain, and loss of function. A model of carrageenan-induced paw edema (CIE) is commonly utilized to test anti-inflammatory substances. Based on the ability of Lepisanthes alata (LA), a tropical plant that is rich in phytochemicals like polyphenols, this study assessed the optimal dose and the health benefits of LA in rats that had been induced with carrageenan to develop paw swelling. Methods: Twenty-four male Wistar rats were divided into four groups to which carrageenan was administered, after which, distilled water at oral dose (C + DW), sodium diclofenac 25 mg/kg (C + DS), LA extract in 250 mg/kg (C + LA250), and 500 mg/kg (C + LA500) was given, respectively. Paw edema was assessed in 24 h. Pain was assessed using the Rat Grimace Scale (RGS), cytokines, antioxidant activity, and tissue changes. Results: LA at 250 and 500 mg/kg significantly decreased paw edema and inflammatory markers in the results of both studies. Remarkably, LA 250 mg/kg significantly decreased RGS scores as well as IL-1β, TNF-α, and histological inflammation but had a positive effect on T-SOD levels. Conclusions: LA extract, especially at 250 mg/kg, shows potent anti-inflammatory, analgesic, and antioxidant properties in CIE rats. Full article

Background: Crystalline glucosamine sulfate (cGS) claims to be a stabilized form of glucosamine sulfate with a defined crystalline structure intended to enhance chemical stability. It is proposed to offer pharmacokinetic advantages over regular glucosamine sulfate (rGS) which is stabilized with potassium or sodium chloride. However, comparative human bioavailability data are limited. Since both forms dissociate in gastric fluid into constituent ions, the impact of cGS formulation on absorption remains uncertain. This pilot study aimed to compare the bioavailability of cGS and rGS using a randomized, double-blind, crossover design. Methods: Ten healthy adults received a single 1500 mg oral dose of either cGS or rGS with a 7-day washout between interventions. Capillary blood samples were collected over 24 h. Glucosamine and its metabolite concentrations were quantified by Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS), and pharmacokinetic parameters—including maximum concentration (C_max), time to reach C_max (T_max), and area under the curve (AUC)—were calculated. Results: Mean AUC_0–24, C_max, T_max, and T_½ values for glucosamine and glucosamine-6-sulfate (GlcN-6-S) were comparable between cGS and rGS. Although the AUC_0–24 for glucosamine was modestly higher with rGS (18,300 ng·h/mL) than with cGS (12,900 ng·h/mL), the difference was not statistically significant (p = 0.136). GlcN-6-S exposure was also similar between formulations (rGS: 50,700 ng·h/mL; cGS: 50,600 ng·h/mL), with a geometric mean ratio of 1.39, a delayed T_max (6–8 h) and longer half-life, consistent with its role as a downstream metabolite. N-acetylglucosamine levels remained stable, indicating potential homeostatic regulation. Conclusions: This pilot study found no significant pharmacokinetic advantage of cGS over rGS. These preliminary findings challenge claims of cGS’ pharmacokinetic superiority, although the small sample size limits definitive conclusions. Larger, adequately powered studies are needed to confirm these results. Full article

Yeasts possess a range of environmental adaptations that allow them to colonize soil and sand. They can circulate seasonally between different components of lake ecosystems, including beach sand, water, and the coastal phyllosphere. The accumulation of people on beaches promotes the development and transmission of yeasts, posing an increasing sanitary and epidemiological risk. The aim of this study was to determine the species and quantitative composition of potentially pathogenic and pathogenic yeasts for humans present in the sand of supervised and unsupervised beaches along the shores of lakes in the city of Olsztyn (northeastern Poland). The study material consisted of sand samples collected during two summer seasons (2019; 2020) from 12 research sites on sandy beaches of four lakes located within the administrative boundaries of Olsztyn. Standard isolation and identification methods used in diagnostic mycological laboratories were applied and are described in detail in the following sections of this study. A total of 259 yeast isolates (264, counting species in two-species isolates separately) belonging to 62 species representing 47 genera were obtained during the study. Among all the isolates, five were identified as mixed (two species from a single colony). Eight isolated species were classified into biosafety level 2 (BSL-2) and risk group 2 (RG-2). The highest average number of viable yeast cells was found in sand samples collected in July 2019 (5.56 × 10² CFU/g), August, and September 2020 (1.03 × 10³ CFU/g and 1.94 × 10³ CFU/g, respectively). The lowest concentrations were in samples collected in April, September, and October 2019, and October 2020 (1.48 × 10² CFU/g, 1.47 × 10² CFU/g, 1.40 × 10² CFU/g, and 1.40 × 10² CFU/g, respectively). The results indicate sand contamination with yeasts that may pose etiological factors for human mycoses. In light of these findings, continuous sanitary-epidemiological monitoring of beach sand and further studies on its mycological cleanliness are warranted, along with actions leading to appropriate legal regulations. Full article

Asia minor bluegrass (Polypogon fugax), a widespread Poaceae weed, exhibits broad tolerance to abiotic stresses. Validated reference genes (RGs) for reliable RT-qPCR normalization in this ecologically and agriculturally significant species remain unidentified. This study identified eight candidate RGs using transcriptome data from seedling tissues. We assessed the expression stability of these eight RGs across various abiotic stresses and developmental stages using Delta Ct, BestKeeper, geNorm, and NormFinder algorithms. A comprehensive stability ranking was generated using RefFinder, with validation performed using the target genes COR413 and P5CS. Results identified EIF4A and TUB as the optimal RG combination for normalizing gene expression during heat stress, cold stress, and growth stages. EIF4A and ACT were most stable under drought stress, EIF4A and 28S under salt stress, and EIF4A and EF-1 under cadmium (Cd) stress. Furthermore, EIF4A and UBQ demonstrated optimal stability under herbicide stress. Additionally, application of validated RGs revealed higher acetyl-CoA carboxylase gene (ACCase) expression in one herbicide-resistant population, suggesting target-site gene overexpression contributes to resistance. This work presents the first systematic evaluation of RGs in P. fugax. The identified stable RGs provide essential tools for future gene expression studies on growth and abiotic stress responses in this species, facilitating deeper insights into the molecular basis of its weediness and adaptability. Full article

Azole resistance in dermatophytes, particularly Trichophyton indotineae, has become a growing global concern. Current antifungal susceptibility testing protocols (EUCAST, CLSI) have limitations in reproducibility and sensitivity. This study aimed to evaluate how medium composition, incubation temperature, and spore concentration influence itraconazole susceptibility testing across various dermatophyte species. Thirty-eight clinical isolates representing Trichophyton, Microsporum, and Epidermophyton species were tested using a microplate laser nephelometry system (MLN). IC₅₀ values for itraconazole were determined in three different media (Sabouraud glucose (SG), RPMI-based (RG), and RG supplemented with casein (RGC)) at 28 °C and 34 °C. Effects of spore concentration on growth dynamics and lag phase were also analyzed. SG medium provided clear phenotypic separation between resistant and sensitive isolates. In contrast, RG and RGC showed overlapping IC₅₀ values. Lower spore concentrations revealed underlying growth differences, which were masked at higher inoculum levels. Temperature and media composition significantly affected IC₅₀ outcomes. Genotypic analysis confirmed resistance-associated Erg11B point mutations and genomic amplifications in T. indotineae, particularly in combination with Erg1 mutations, forming distinct subpopulations. SG medium combined with reduced spore concentrations offered improved differentiation of resistant versus sensitive strains. These findings support the development of more accurate susceptibility testing protocols and highlight the need to establish species-specific ECOFF values for dermatophytes. Full article

Accurate gene expression quantification using reverse transcription quantitative PCR (RT-qPCR) requires stable reference genes (RGs) for reliable normalization. However, few studies have systematically identified RGs suitable for simultaneous high salt, alkaline, and high-temperature conditions. This study addresses this gap by evaluating the stability of eight candidate RGs in the anaerobic halophilic alkalithermophile Natranaerobius thermophilus JW/NM-WN-LF^T under combined salt, alkali, and thermal stresses. The stability of these candidate RGs was assessed using five statistical algorithms: Delta CT, geNorm, NormFinder, BestKeeper, and RefFinder. Results indicated that recA exhibited the highest expression stability across all tested conditions and proved adequate as a single RG for normalization in both RT-qPCR and droplet digital PCR (ddPCR) assays. Furthermore, recA alone or combined with other RGs (sigA, rsmH) effectively normalized the expression of seven stress-response genes (proX, opuAC, mnhE, nhaC, trkH, ducA, and pimT). This work represents the first systematic validation of RGs under polyextreme stress conditions, providing essential guidelines for future gene expression studies in extreme environments and aiding research on microbial adaptation mechanisms in halophilic, alkaliphilic, and thermophilic microorganisms. Full article

Background: Lung cancer has the highest morbidity and mortality of all tumors, and the development of TKI drugs targeting EGFR activating mutations has brought lung cancer treatment into the targeted era. In view of their low efficacy and susceptibility to drug resistance, there is an urgent need to find strategies to increase their efficacy and reduce the incidence of drug resistance. Methods: In this study, we examined the distribution and probability of EGFR mutations in non-small cell lung cancer patients in the cBioPortal database and compared the survival prognosis of patients with normal and abnormal EGFR, NSCLC patients treated with and without TKI, and NSCLC patients with different EGFR gene copy numbers. We established a mouse lung cancer model and examined the histomorphological characteristics of lung tissues via hematoxylin and eosin staining. Additionally, changes in the copy number of the EGFR gene and its protein expression levels were detected using RT-qPCR and Western blotting. Furthermore, we quantified the concentration of the EGFR protein using ELISA. Results: We found no significant advantage of EGFR-TKI therapy over first-line chemotherapeutic agents in patients with EGFR-abnormal NSCLC. The reason for this may be related to the abnormal EGFR gene copy number; the higher the copy number increases, the worse the survival prognosis of the patients. In molecular biology experiments, we demonstrated that ginsenoside Rg3 down-regulated the copy number of 18, 19, 20, and 21 exons and protein expression of EGFR in lung adenocarcinoma cells. The results of in vivo pharmacodynamic assays confirmed that sequential administration of ginsenoside Rg3 with TKI drugs could achieve a gainful complementary effect. Conclusions: Ginsenoside Rg3 down-regulates the copy number of EGFR important exons in EGFR-mutant cells of lung adenocarcinoma and reduces EGFR protein expression, thus providing a high gainful complementary effect in combination with EGFR-TKI. Full article

Abies nebrodensis L. is a critically endangered conifer endemic to Sicily (Italy). Its residual population is confined to the Madonie mountain range under challenging climatological conditions. Despite the good adaptation shown by the relict population to the environmental conditions occurring in its habitat, Abies nebrodensis is subject to a series of threats, including climate change. Effective conservation strategies require reliable and versatile methods for monitoring its health status. Combining high-resolution remote sensing data with reanalysis of climatological datasets, this study aimed to identify correlations between vegetation indices (NDVI, GreenDVI, and EVI) and key climatological variables (temperature and precipitation) using advanced machine learning techniques. High-resolution RGB (Red, Green, Blue) and IrRG (infrared, Red, Green) maps were used to delineate tree crowns and extract statistics related to the selected vegetation indices. The results of phytosanitary inspections and multispectral analyses showed that the microclimatic conditions at the site level influence both the impact of crown disorders and tree physiology in terms of water content and photosynthetic activity. Hence, the correlation between the phytosanitary inspection results and vegetation indices suggests that multispectral techniques with drones can provide reliable indications of the health status of Abies nebrodensis trees. The findings of this study provide significant insights into the influence of environmental stress on Abies nebrodensis and offer a basis for developing new monitoring procedures that could assist in managing conservation measures. Full article

This study reports the use of single-cell RNA sequencing to evaluate B cells in the peripheral blood mononuclear cells (PBMCs) and intrathyroidal blood mononuclear cells of patients with Graves’ disease (GD) undergoing thyroidectomy. These cells were stimulated with overlapping peptides of thyroid autoantigens, including thyroid-stimulating hormone receptor (TSHR), thyroglobulin (Tg), and thyroid peroxidase (TPO). In PBMCs, naive B cells are characterized by IL6 and CXCR5, whereas memory B cells express IGHG1, IGHG2, and CD74. HLA-DMA, HLA-DRB1, IGHG, IGHM, CD74, CD79A, and MS4A1 expression increased in peptide-stimulated naive and memory B cells compared to those in the controls. Thyroid naive B cells are characterized by CD40 and TNFRSF13C, whereas memory B cells express IGHM, CD79A, and MS4A1. Thyroid B cells showed higher DUSP1, DUSP2, CD69, FOSB, RGS1, and immunoglobulin gene expression than control PBMCs and thyroid cells. B-cell receptor analysis revealed frequent IGHV3-23 and IGHV4-34 usage in controls, whereas IGHV4-34/IGHJ4 expression was increased in TSHR-stimulated groups. We concluded that B-cell responses to TSHR, Tg, and TPO differed and that changes in B-cell reactivity also occurred in PBMCs and the thyroid. Additionally, IGHV3-23 and IGHV4-34 may be associated with autoantibody production in GD. Full article

The relationship between protein structure and function is intrinsically interconnected, as the structure of a protein directly determines its functional properties. To investigate the effects of temperature and pressure on protein function, this study employed ethyl carbamate (EC) hydrolase as a model food enzyme and conducted molecular dynamics (MD) simulations under varying temperature and pressure levels to elucidate its structure–function relationship. By systematically analyzing the dynamic changes in root mean square deviation (RMSD), root mean square fluctuation (RMSF), radius of gyration (Rg), solvent accessible surface area (SASA), hydrogen bonding, catalytic pocket conformation, and packing density under different temperature and pressure conditions, we revealed the structural adaptability of EC hydrolase. Furthermore, we analyzed the characteristics of EC hydrolase using molecular dynamics simulations with temperature and pressure levels, as well as conformational bias-based computer-aided engineering, providing both theoretical and experimental foundation for the adaptability mechanisms of enzymes under extreme conditions. Full article

High-voltage unipolar square wave pulsed electric fields (PEFs) can cause cell membrane rupture and cell death during a process termed irreversible electroporation (IRE). PEF effects are influenced by pulse parameters like number of pulses (NP), voltage (PV), width (PW), and interval (PI). This study systematically evaluates their effects on the conductivity and relative conductivity changes between untreated and PEF-treated regions of potato tissue across a frequency range of 1 Hz to 5 MHz by means of electrochemical impedance spectroscopy (EIS), using a custom-made four-point EIS probe with RG58/U coaxial cables. Potatoes were chosen as a plant-based PEF model to reduce animal experiments and untreated tissue showed minimal conductivity variation across regions. Relative conductivity changes were maximal at 1000 Hz. At 1000 Hz, significant conductivity differences between untreated and PEF-treated regions were observed from PV = 200 V, NP = 10, PW = 10 µs, and PI = 50 ms onwards (most significant changes occurred for PV = 700 V; NP = 70; PW = 70 µs; PI = 250 ms and 500 ms). Our results may be beneficial for multiphysics modelling of IRE with specific electrical properties, conductivity mapping with optimal contrast—such as in electrical impedance tomography—and development of IRE procedures. Full article

This study examines a Mei-Yu rainfall event using rain gauges (RG) and OTT Parsivel disdrometers to observe precipitation characteristics and raindrop size distributions (RSD), with comparisons made against Weather Research and Forecasting (WRF) model simulations. Results show that Parsivel-derived rain rates (RR) are slightly underestimated relative to RG measurements. Both observations and simulations identify 1–3 mm raindrops as the dominant precipitation contributors, though the model overestimates small and large drop contributions. At low RR, decreased small-drop and increased large-drop concentrations cause corresponding leftward and rightward RSD shifts with decreasing altitude—a pattern well captured by simulations. However, at elevated rainfall rates, the simulated concentration of large raindrops shows no significant increase, resulting in negligible rightward shifting of RSD in the model outputs. Autoconversion from cloud droplets to raindrops (ATcr), collision and breakup between raindrops (AGrr), ice melting (MLir), and evaporation of raindrops (VDrv) contribute more to the number density of raindrops. At 0.1 < RR < 1 mm·h⁻¹, ATcr dominates, while VDrv peaks in this intensity range before decreasing. At higher intensities (RR > 20 mm·h⁻¹), AGrr contributes most, followed by MLir. When the RR is high enough, the breakup of raindrops plays a more important role than collision, leading to a decrease in the number density of raindrops. The overestimation of raindrop breakup from the numerical parameterization may be one of the reasons why the RSD does not shift significantly to the right toward the surface under the heavy RR grade. The RSD near the surface varies with the RR and characterizes surface precipitation well. Toward the surface, ATcr and VDrv, but not AGrr, become similar when precipitation approaches. Full article

Microbial fermentation technology has emerged as a pivotal approach for enhancing ginseng efficacy through the transformation of active ingredient molecular structures. This paper reviews the impact of microbial fermentation on the structure–activity relationship of ginseng bioactive compounds and advances in its application. Bibliometric analysis indicates that Panax species (Panax ginseng, Panax notoginseng) are primarily fermented using lactic acid bacteria and Aspergillus spp., with research predominantly focused on conversion efficiency to rare ginsenosides (Compound K, Rg3, and Rh2). Specifically, this review details the biotransformation pathways of these rare ginsenosides and the resultant bioactivity enhancements. Additionally, it summarizes the effects of other microorganisms, such as fungal fruiting bodies, on additional ginseng constituents like polysaccharides and polyphenols. Microbial fermentation has been successfully implemented in functional products, including ginseng vinegar, wine, and fermented milk. This review subsequently examines these applications, emphasizing fermentation’s potential to enhance product functionality. However, challenges remain in strain screening, process standardization, and analysis of multi-component synergistic mechanisms. In summary, this review synthesizes recent advancements in understanding the mechanisms of microbial fermentation on ginseng and its translational applications in functional foods and pharmaceuticals. Full article

Background and Objectives: The common complaints of head and neck cancer patients receiving concurrent chemo-radiotherapy (CCRT) are dry mouth, dysphagia, trismus, hoarseness, sore throat, and oral mucosal damage, which result in retained secretions and difficult expectoration. We aimed to investigate the effect of adjuvant respiratory therapy on secretion expectoration and treatment completion in patients with head and neck cancer receiving CCRT. Materials and Methods: From November 2016 to May 2018, 56 head and neck cancer patients were recruited retrospectively, and according to their respiratory therapy in the medical record, were divided into the control group (CG, n = 27) or the research group (RG, n = 29). In the CG, the patients were treated via the teaching of routine breathing exercises and expel techniques, while patients in the RG were treated with the inhalation of a ß-agonist bronchodilator agent five times each week, in addition to the standard treatment administered in the CG. Results: The total completion rate of treatment was significantly higher in the RG (21 patients) compared with the CG (12 patients) (72.4% vs. 44.4%, p < 0.05). After therapy, the rates of clinical symptoms were significantly increased in the RG compared with the CG, including smooth expectoration (76.2% vs. 75.0%), decreased secretions (61.9% vs. 58.3%), reduced viscosity of secretions (66.7% vs. 58.3%), lower cough frequency (71.4% vs. 50.0%), improved sore throat (52.4% vs. 41.7%), and swallowing function (52.4% vs. 50.0%). The continuation of chemo-radiotherapy without disruption was higher in the RG than it was in the CG (66.7% vs. 50.0%). There was no significant difference in adverse effects between the two groups. Conclusions: Adjuvant respiratory therapy not only improves secretion expectoration, but also reduces side effects, thus promoting the completion of the CCRT schedule in patients with head and neck cancer. Full article

We develop a symmetry-based variational theory that shows the coarse-grained balance of work inflow to heat outflow in a driven, dissipative system relaxed to the golden ratio. Two order-2 Möbius transformations—a self-dual flip and a self-similar shift—generate a discrete non-abelian subgroup of $\mathrm{PGL}(2,\mathbb{Q}\left(\sqrt{5}\right))$. Requiring any smooth, strictly convex Lyapunov functional to be invariant under both maps enforces a single non-equilibrium fixed point: the golden mean. We confirm this result by (i) a gradient-flow partial-differential equation, (ii) a birth–death Markov chain whose continuum limit is Fokker–Planck, (iii) a Martin–Siggia–Rose field theory, and (iv) exact Ward identities that protect the fixed point against noise. Microscopic kinetics merely set the approach rate; three parameter-free invariants emerge: a $62\%:38\%$ split between entropy production and useful power, an RG-invariant diffusion coefficient linking relaxation time and correlation length ${\mathcal{D}}_{\alpha }={\xi }^z/\tau$, and a $\vartheta ={45}^{\circ }$ eigen-angle that maps to the golden logarithmic spiral. The same dual symmetry underlies scaling laws in rotating turbulence, plant phyllotaxis, cortical avalanches, quantum critical metals, and even de-Sitter cosmology, providing a falsifiable, unifying principle for pattern formation far from equilibrium. Full article