Search Results (44,613)

Background: Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) can develop as post-vaccination syndrome (PVS) or Post-Acute Sequelae of SARS-CoV-2 infection (PASC). In our prior retrospective study, most patients with PVS who developed ME/CFS had vitamin D insufficiency or deficiency. We evaluated the efficacy of vitamin D replacement therapy guidance for ME/CFS symptom improvement in patients with vitamin D insufficiency or deficiency. Methods: This open-label randomized controlled trial enrolled 91 participants with ME/CFS as PVS or PASC and serum 25(OH) vitamin D < 30 ng/mL across five clinical sites. Participants were randomized 1:1 to intervention (active vitamin D preparation plus vitamin D replacement therapy guidance: 25 μg daily supplementation, dietary counseling, sun exposure, and exercise) or control (active vitamin D preparation alone) for 12 weeks. The primary endpoint was the change in ME/CFS symptom count from screening to Week 12. Results: Mean symptom change was −6.7 in the intervention group versus −1.2 in the control group (between-group difference −5.6; 95% CI: −7.2, −3.9; p < 0.001). Serum 25(OH) vitamin D improved from 18.6 to 27.1 ng/mL in the intervention group, while the control group showed a decreasing trend (between-group difference 10.2 ng/mL; 95% CI: 7.9, 12.5). Achievement of <8 symptoms (i.e., no longer meeting ME/CFS diagnostic criteria) was significantly higher in the intervention group, with 16 participants achieving this threshold compared to 1 in the control group (p < 0.001). Subgroup analyses showed consistent benefit in both PVS (n = 56) and PASC (n = 29) cohorts. Conclusions: Vitamin D replacement therapy guidance significantly reduced ME/CFS symptoms along with improvement of serum 25(OH) vitamin D levels in patients with vitamin D insufficiency or deficiency who developed ME/CFS as PVS or PASC. Full article

Introduction: Plasma is the standard biological fluid used in pharmacokinetic (PK) studies and therapeutic drug monitoring (TDM) of pyrazinamide, a key antituberculosis (TB) drug. This study described the PK of pyrazinamide in saliva and investigated whether saliva could serve as an alternative matrix for pyrazinamide PK evaluations. Methods: Fifteen adult Tanzanian TB patients in the intensive treatment phase participated in a descriptive PK study. Time-matched saliva (stimulated using a Salivette^® with citric acid) and plasma samples were collected at multiple intervals up to 24 h after drug intake. Pyrazinamide concentrations were measured using validated HPLC methods, exposure measures were assessed, and predictive performance for salivary concentrations was determined. Results: Salivary exposure to pyrazinamide (AUC_0–24h: 230 h·mg/L; C_max: 28.6 mg/L) was lower than plasma exposure (AUC_0–24h: 377 h·mg/L; C_max: 36.4 mg/L, p < 0.001), but T_max was similar (median 2.0 h, p = 0.893). A saliva/plasma ratio of 0.59 was assessed, and a reciprocal conversion factor of 1.68 allowed for reasonably accurate (bias 5.8%) but imprecise (imprecision 24.3%) plasma concentration predictions from saliva. Use of a conversion factor of 1.49, based on more stable saliva/plasma concentration ratios for samples between 2 and 6 h post-dose, resulted in a bias of 0.74% and imprecision of 17.7% for predicting plasma concentrations from salivary concentrations in the 2–6 h interval. Conclusions: The exposure to pyrazinamide in saliva is relatively high. Salivary measurement of pyrazinamide can be used as a semi-quantitative predictor of pyrazinamide plasma concentrations. Full article

Background/Objectives: Oleuropein is a bioactive phenolic compound from olive leaves with antimicrobial and antioxidant activity. This study aimed to develop a sprayable oral gel containing an oleuropein-rich aqueous extract and to evaluate its pharmaceutical performance antimicrobial efficacy and in ovo biological response. Methods: Oleuropein content was quantified using a validated chromatographic method. Polymeric systems were screened to select an optimized sprayable formulation. Physicochemical stability, dose uniformity, and antimicrobial activity against major cariogenic bacteria were evaluated. In ovo biological evaluation was conducted using the chick chorioallantoic membrane angiogenesis model together with histopathological examination of embryonic heart and liver tissues. Results: Oleuropein content was determined as 288.6 µg/mL in the olive leaf extract and 255.1 µg/mL in the final formulation. The optimized oral spray showed stable physicochemical properties, with pH maintained at 6.90 ± 0.02 and no relevant changes in viscosity during storage. The mean delivered dose per actuation was 0.128 ± 0.015 g, corresponding to 32.6 µg oleuropein per spray. The formulation exhibited inhibitory activity against all tested cariogenic microorganisms, with MIC values ranging from 13.3 to 170.7 µg/mL and MBC values generally two-fold higher. In the CAM assay, significant concentration- and time-dependent antiangiogenic effects were observed after 24–48 h at moderate and higher concentrations. Histopathological evaluation revealed dose-dependent acute degenerative and congestive changes in heart and liver tissues without evidence of fibrosis or steatosis. Conclusions: The oleuropein-based sprayable oral gel is a promising localized delivery system with adequate stability dose uniformity and antimicrobial efficacy. In ovo findings provide a conservative assessment of systemic exposure and support further development for oral biofilm and caries-related applications. Full article

Background: Osteoporosis is a prevalent metabolic bone disorder characterized by reduced bone mineral density (BMD) and increased fracture risk, particularly among older adults. While individual medications have been implicated in bone loss, the cumulative impact of polypharmacy on skeletal health remains underexplored. Methods: This cross-sectional study included 1155 geriatric inpatients undergoing routine bone mineral density assessment. Medication use, demographic characteristics, and clinical variables were extracted from electronic medical records. BMD at the lumbar spine (L1–L4) and total hip was measured using dual-energy X-ray absorptiometry (DXA). Unadjusted analyses and multivariable linear regression models were used to examine associations between medication use, polypharmacy (defined as the use of ≥5 medications), and BMD, adjusting for age, sex, body mass index, and relevant clinical covariates. Results: The mean age of the study population was 85.0 ± 7.1 years, and 80.1% were female. Polypharmacy was present in 64.5% of patients. In medication-specific analyses, thyroid hormone use was associated with lower lumbar spine BMD (p = 0.032), and concomitant use of diuretics and proton pump inhibitors was associated with lower hip BMD (p = 0.049). Steroid use showed a marginally non-significant correlation with reduced BMD (p = 0.057). Polypharmacy was associated with lower lumbar spine BMD (p = 0.022), whereas no significant association was observed with hip BMD. Increasing age was consistently associated with lower BMD across skeletal sites (p < 0.001). Conclusions: In this geriatric inpatient cohort, polypharmacy and selected medication classes were associated with lower bone mineral density, particularly at the lumbar spine. Given the cross-sectional design, these findings reflect associations rather than causal relationships and may partly capture underlying multimorbidity and clinical complexity. Consideration of medication burden may be relevant when evaluating bone health in older adults. Polypharmacy is increasingly common in older adults and may contribute to bone fragility. In this cohort of 1155 geriatric inpatients, multiple medications and certain drug classes were associated with lower bone mineral density, particularly in the spine. These findings suggest that medication burden should be considered when evaluating osteoporosis risk in aging populations. Full article

Background: Immune checkpoint blockade (ICB) has revolutionized lung adenocarcinoma (LUAD) therapy, yet predictive bio-markers remain suboptimal. We hypothesized that BCL2A1 expression in CD8⁺ T cells may reflect immune endurance and complement PD-L1 in predicting ICB response. Methods: Integrating bulk and single-cell RNA-seq across multiple LUAD cohorts, this study performed differential expression, survival, and pathway analyses in a discovery cohort (n = 60) and validated findings across five independent cohorts (n = 126). Results: Single-cell profiling identified BCL2A1 enrichment in tissue-resident memory and proliferating subsets that appeared preferentially expanded in responders; cell–cell communication analysis revealed that BCL2A1^high CD8⁺ T cells exhibited significantly enhanced outgoing signaling capacity (p = 0.0278), with proliferating subsets serving as intra-CD8⁺ coordination hubs and MIF pathway interactions achieving the highest intensity among all axes examined. BCL2A1 was significantly upregulated in responders (FDR < 0.05) and associated with improved ICB survival (HR = 0.43, p < 0.05), but not in non-ICB settings, suggesting treatment-specific prognostic relevance. A tri-marker model integrating BCL2A1, PD-L1 (CD274), and a 27-gene HOT score demonstrated favorable predictive performance (AUC = 0.826 discovery; macro-AUC = 0.774 validation), outperforming PD-L1 alone (AUC = 0.706) and established signatures including TIDE, IPS, TIS, and IFNG. Cross-platform simulations suggested high reproducibility (ρ = 0.982–0.993). Conclusions: These findings suggest BCL2A1 may serve as a bio-marker of CD8⁺ T-cell survival and enhanced intercellular coordination, and its integration with PD-L1 and immune activation markers may yield a reproducible ICB response predictor, pending clinical validation. Full article

Background/Objectives:This study evaluated how variations in emulsion composition influence the viability of a probiotic strain Lactobacillus rhamnosus GG within biphasic systems, as well as the overall stability of the resulting formulations. Methods:Eight biphasic emulsions were prepared, each in two versions—with and without a preservative—and subsequently analysed for pH, FTIR spectroscopy, and emulsion type. The viability of L. rhamnosus GG in each formulation was determined using the plate count method. This method is regarded as the reference technique for the quantitative determination of viable bacteria, expressed as colony-forming units (CFUs). Results: The pH indicated that an emulsion with a pH of 4.65 provides the most favorable conditions for L. rhamnosus GG survival, as values below pH 6 promote its proliferation. This acidity aligns with the natural pH of human skin, although it falls slightly below the recommended 4–5 range for topical formulations. FTIR analysis confirmed the structural stability of the emulsions and revealed spectral shifts attributable to the presence of the bacteria. The spectra remained largely consistent throughout the study period, demonstrating good temporal stability. Conclusions: Microbiological evaluation showed that all produced formulations supported bacterial growth, the presence of the preservative did not inhibit viability of L. rhamnosus GG, corroborating findings from an independent assessment. All emulsions were classified as O/W systems, due to the high water content which is advantageous for microbial viability. Furthermore, O/W emulsions are user-friendly, easy to remove, limit the penetration of the active component into deeper skin layers, supporting their suitability for probiotic-based topical applications. Full article

The rapid expansion of greenhouse agriculture demands sustainable strategies to maintain soil health and productivity from the outset. Priestia megaterium, a plant growth-promoting rhizobacterium (PGPR), has shown promise in improving plant growth and soil nutrient availability, but its efficacy in newly established greenhouse systems, where the soil microbiome is still developing, remains underexplored. This study evaluated the impact of P. megaterium inoculation on cucumber growth, soil nutrient bioavailability, and soil microbial communities in a greenhouse with only two years of operation. A two-year experiment was conducted with conventional fertilization as the control and P. megaterium inoculation (7.0 × 10⁸ cfu mL⁻¹) at different rates (37.5, 75, 150, and 300 L ha⁻¹) and timings. Soil and plant nutrient content were measured, and microbial communities were analyzed through 16S rRNA sequencing and co-occurrence network analysis. Results showed that applying P. megaterium at 75 L ha⁻¹ during seedling transplantation significantly increased soil available phosphorus (AP) by 11.64–26.48% and available potassium (AK) by 11.27–47.31% compared to the control, while enhancing cucumber yield by 6.71–9.28%. The inoculant also increased soil bacterial diversity, enriched beneficial genera such as Lysobacter, Pseudomonas, and Flavobacterium, and reduced the abundance of Xanthomonas. Furthermore, P. megaterium application promoted a more complex and stable bacterial network, with higher connectivity and modularity. These findings suggest that P. megaterium is a viable strategy for enhancing soil health and productivity in newly established greenhouse systems, offering an environmentally sustainable alternative to traditional fertilization methods. Full article

The development of oxidation-resistant and regenerable materials remains a major challenge for mercury removal from contaminated waters and industrial effluents. In this study, a zwitterionic mesoporous silica gel functionalized with L-cysteine (SG-3PS-Cys) was synthesized, where the thiol group is covalently anchored to the silica framework, preventing oxidative degradation while preserving –NH₃⁺ and –COO⁻ groups for Hg(II) coordination. Spectroscopic analyses (FTIR, XPS, and ¹³C NMR) confirmed the formation of a stable, thiol-free binding environment in which mercury interacts through carboxylate oxygen atoms, electrostatically stabilized by neighboring ammonium groups. The material exhibited a high surface area (134 m² g⁻¹) and uniform mesoporosity (9.8 nm), achieving a maximum Hg(II) uptake of 82.7 mg g⁻¹ at pH 3 with rapid kinetics and cooperative S-type isotherms. The adsorbent retained 72% of its capacity after five regeneration cycles and maintained 38.7% selectivity toward Hg(II) in multicomponent solutions. DFT-based surface energy distribution analysis supported the zwitterionic coordination mechanism, revealing energetically homogeneous and high-affinity binding domains. Beyond its chemical stability, the material introduces a sustainable route for mercury remediation, linking surface energy, electrostatic effects, and porosity to achieve durable performance under acidic and complex aqueous conditions. These findings provide a mechanistic and design framework for the next generation of non-thiol adsorbents capable of selective and reusable Hg(II) removal in environmentally relevant scenarios. Full article

Monitoring the soil–plant system in forest ecosystems is crucial for preserving their ecological functions and services. This study assessed carbon and nitrogen stable isotopes and ecoenzymatic stoichiometry as suitable indicators for characterizing the soil–plant system as a functional unit of ecological processes. To this end, in June 2021 six plots (1 m² each) were selected in two typical Mediterranean forest ecotypes: a coastal stone pine forest (Pinus pinea L., PF) and a meso-hygrophilous broadleaf forest (RV). Soil samples (0–15 and 15–30 cm depth) and litter samples (40 × 40 cm) were collected and characterized in terms of physical, chemical and biochemical properties. t-tests revealed significant differences between RV and PF, indicating distinct microbial nutrient acquisition strategies. The higher C:N ratio in PF suggested lower litter quality and greater recalcitrance to microbial decomposition. Consistently, RV showed a more pronounced ¹³C and ¹⁵N enrichment from litter to SOM down to a 30 cm depth, confirming faster organic matter decomposition and mineralization. Enzyme activity patterns supported these findings. The higher β-glucosidase and butyrate esterase activities in RV reflected its greater microbial potential to activate biogeochemical cycles. Both forests exhibited a higher microbial demand for C and P than for N to maintain ecological stoichiometric balance, with stronger C limitation at the surface and P limitation in the subsoil, particularly in RV soil. This integrated monitoring approach provides insights into nutrient cycling and ecosystem resilience and offers tools to evaluate ecosystem functionality under changing environmental conditions, supporting sustainable forest management. Full article

Background/Objectives: Dalbavancin is approved for pediatric acute bacterial skin and skin structure infections (ABSSSIs), yet real-world practice frequently necessitates off-label use for deep-seated infections requiring prolonged suppression. While adult data support therapeutic drug monitoring (TDM)-guided maintenance, the pediatric evidence for repeated-dose pharmacokinetics (PK) is limited. We evaluated the efficacy, safety, multi-dose PK, and pharmacoeconomic impact of dalbavancin in a complex pediatric cohort. Methods: A retrospective study (2023–2025) of enrolled patients < 18 years treated with dalbavancin. A subgroup receiving ≥3 doses underwent PK analysis to assess concentration decay against conservative efficacy targets (4 and 8 mg/L). A pharmacoeconomic analysis compared resource utilization against the standard of care. Results: Sixteen patients (median age 12) were included, primarily treated for Staphylococcus aureus (S. aureus) osteoarticular infections (75%), and frequently device-associated (66.7%). Clinical success was 93.8% (15/16) with no adverse events. A PK analysis (n = 9; 78 samples) ruled out dangerous accumulation but revealed a significant concentration drop at week 4 (mean 6.06 mg/L; p = 0.005). Logistic regression identified the time since the previous dose as the sole predictor of sub-therapeutic levels, with >50% of the patients dropping below 8 mg/L by the fourth week. An analysis showed median net savings of EUR 3215.84 per patient (p = 0.004). Conclusions: Dalbavancin is effective and cost-saving for complex pediatric infections. However, due to distinct pediatric PK, dosing regimens extrapolated from adults may result in sub-therapeutic concentrations by week 4. We recommend TDM around week 3 to tailor dosing or limiting maintenance intervals to a maximum of 4 weeks. Full article

Post-translational modifications of human serum albumin (HSA) have been described in patients with liver disease. This prospective cohort study aimed to characterize HSA microheterogeneity in hospitalized patients with alcohol-associated hepatitis (AH) and investigate its clinical relevance. We analyzed HSA isoforms by mass spectrometry in 49 patients with AH (at admission and day 14) and 20 healthy controls. Survival at 30, 90, and 365 days was assessed. Differences in HSA isoform abundance were compared between controls and AH patients, as well as between 90-day survivors and non-survivors. AH patients (69% male, median age 53 years) exhibited a significantly different HSA form profile compared to controls, with a lower amount of native HSA and higher oxidized forms. Native HSA negatively correlated with total HSA concentration (R = −0.47, p < 0.001). The relative amount of native HSA increased non-significantly from admission to day 14, but its estimated concentration increased significantly (8.8 vs. 12.0 g/L, p = 0.005). There were no significant differences in HSA forms between 90-day survivors and non-survivors at admission or day 14. Patients with AH exhibit extensive post-translational modifications of HSA compared to healthy individuals. While HSA forms changed during early hospitalization, they did not significantly correlate with short-term mortality in this cohort. Full article

Chitinases catalyze the hydrolysis of β-1,4-glycosidic bonds in chitin, a structural biopolymer synthesized by numerous organisms. Although these enzymes have been widely investigated, studies focusing on insect-derived chitinases remain limited. In this study, three recombinant chitinases from the leaf-cutter ant Atta sexdens were cloned, expressed in Pichia pastoris, and biochemically characterized. The enzymes-AsChtII-C2B3 (one catalytic and three chitin-binding domains), AsChtII-C3C4 (two catalytic domains), and AsChtII-C5B1 (one catalytic and one binding domain), exhibited optimal activity at pH 4–5 and 50 °C using colloidal chitin as substrate. Chitinase activity on colloidal α-chitin was confirmed by ¹H NMR (proton nuclear magnetic resonance) spectroscopy, revealing GlcNAc concentrations of 0.41, 0.48, and 0.56 mmol L⁻¹ for AsChtII-C3C4, AsChtII-C2B3, and AsChtII-C5B1, respectively. Their antifungal activities were evaluated against the human pathogens Candida albicans and Aspergillus fumigatus, as well as the phytopathogen Lasiodiplodia theobromae. Distinct inhibition profiles were observed: AsChtII-C5B1 (150 µg/mL) showed the highest activity against C. albicans (87.6% inhibition), while AsChtII-C3C4 (25 µg/mL) was most effective against A. fumigatus (60% inhibition). Notably, only AsChtII-C2B3 inhibited L. theobromae growth, inducing severe hyphal deformations observed by scanning electron microscopy (SEM). These findings demonstrate that recombinant A. sexdens chitinases exhibit species-specific antifungal properties, underscoring their potential as biotechnological tools for medical and agricultural applications. Full article

Objectives: This study aimed to evaluate phenotypic associations between preoperative alkaline phosphatase (ALP) levels and clinical characteristics, and explore clinical factors associated for postoperative regrowth in craniofacial fibrous dysplasia. Methods: In this retrospective cohort (2003–2024), 71 surgically treated fibrous dysplasia patients were analyzed. Relationships between preoperative ALP (using age-stratified reference ranges) and key phenotypes (age at surgery, onset age, laterality, lesion type) were assessed via nonparametric tests. Associations with postoperative regrowth were assessed using Mann–Whitney U or Kruskal–Wallis tests for non-normally distributed continuous variables and χ² or Fisher’s exact tests for categorical variables. Results: Preoperative ALP levels significantly correlated with younger surgical age (16–19 vs. ≥19 years: 244.0 vs. 107.0 U/L, p < 0.001), earlier onset (0–16 vs. >16 years: 114.0 vs. 83.0 U/L, p = 0.030), bilateral lesions (176.0 vs. 106.2 U/L, p = 0.006), and polyostotic subtype (polyostotic fibrous dysplasia vs. monostotic fibrous dysplasia: 162.0 vs. 87.5 U/L, p < 0.001). However, neither ALP levels (p = 0.061) nor abnormal ALP rates (p = 0.090) predicted regrowth. Crucially, bilateral lesions were significantly associated with regrowth (83.3% (5/6) vs. 21.5% (14/65); p = 0.005). The overall regrowth rate was 8.5% (6/71). Conclusions: Bilateral lesions demonstrate significant association with postoperative regrowth risk, potentially guiding surveillance intensity. ALP correlates with phenotypic burden but shows limited prognostic utility. These findings, interpreted considering retrospective constraints, warrant validation in larger cohorts. Full article

Background/Objectives: In clinical practice today, platelet concentrates are often used for topical surgical applications. They are biomaterials that can accelerate healing processes associated with oral and maxillofacial surgery as well as in several other clinical applications through the action of growth factors released by platelets at the surgical site. However, in most cases, the exact quantification of the released growth factors is challenging in both the short and long term. The aim of this study was to determine if early platelet activation and degranulation occur during the collection and utilization of platelet-rich plasma (PRP) in the surgical room, where, before its application, PRP undergoes a procedure of gelification via reactions with procoagulant agents. Methods: PRP was prepared from the blood samples of 39 patients following the modified Whitman protocol. The samples were then analyzed at four different time points (1, 6, and 24 h during preparation and clinical application in the surgery room) using flow cytometry and enzyme-linked immunosorbent assays (ELISAs) to investigate the platelet activation/degranulation and TGFβ release in the supernatant (SN) during storage and clinical application. The mean platelet count in the whole blood was 267.5 ± 48.58 × 10³/mL (range: 189–334 × 10³/mL), and the mean concentration was 2925.5 ± 833.37 × 10³/mL (range: 748–3453 × 10³/mL). Results: The activation and degranulation of platelet cells (measured via monoclonal antibodies: CD62p and CD63, respectively) demonstrated a progressive increase at 1 h, 6 h, 24 h, and after gelification. The TGFβ dosage in the supernatant (SN) at different times exhibited a similar trend, with a mean release of 18.36 ng/mL at 1 h, 21.96 ng/mL at 6 h, and 29.45 ng/mL at 24 h. After the gelification of the PRP, a significant reduction was observed, with a value of 15.52 ng/mL. Conclusions: The results reveal that the protocol used for the preparation, storage, and application of the PRP ensures a good-quality hemoderivative and that the platelet concentrate must be applied with the correct timing to support tissue healing processes. Full article

A supplemental description of Lamproglena barbicola Fryer, 1961 is provided based on specimens collected from the gills of Labeobarbus altianalis (Boulenger, 1900) from the Nyando River, Lake Victoria Basin, Kenya, using an integrated approach of scanning electron microscopy (SEM) and molecular analysis (18S, 28S rDNA, and cox1 gene regions). Morphologically, the specimens conform to L. barbicola and closely resemble Lamproglena hoi Dippenaar, Luus-Powell & Roux, 2001; however, SEM revealed a previously undescribed feature on the uniramous antennule in L. barbicola, namely indistinctly three-segmented, tapering from a broad base to the apex, basal segment much longer than distal, comprising 14 setae of varying sizes, ventral laterally, absence of distinctive anterior fringe of setae on the antennule, as well as several characters that differentiate L. barbicola from L. hoi, including 5 setae at the basal endopod of leg one, five cuticular protuberances in the oral region, 19 setae on the basal antennular segment, and 10 setae on the distal segment, with 1 seta on each ramus. The phylogenetic analysis confirms L. barbicola as a sister taxon of L. hoi, supporting their close relationship. The genetic divergence presented as the uncorrected genetic p-distances between L. barbicola and L. hoi are 23.1% and 0.45% for cox1 and 28S rDNA regions, respectively, with observed nucleotide differences of 145 and 3 bp between the sequences, respectively. There was no interspecific variability detected in the 18S rDNA sequences. This study provides novel molecular sequences and the first high-resolution SEM images, which reveal additional taxonomic features for L. barbicola, establishing a robust reference for future identification. Full article