Search Results (2,008)

Background: Microsatellite instability (MSI) is an important biomarker for the diagnosis of Lynch syndrome and for guiding immunotherapy in various solid tumors. Droplet digital PCR (ddPCR) has emerged as a highly sensitive method for detecting MSI, particularly in circulating tumor DNA (ctDNA). This study aimed to evaluate the analytical and clinical performance of a ddPCR assay using three MSI markers (BAT-26, ACVR2A, and DEFB105A/B) in colorectal, gastric, and endometrial cancers. Methods: Formalin-fixed paraffin-embedded (FFPE) samples from 190 patients (83 colorectal, 44 gastric, and 63 endometrial cancers) and 21 plasma samples from patients with metastatic solid tumors were analyzed. MSI status determined by ddPCR was compared with conventional PCR using a pentaplex panel and immunohistochemistry (IHC). Analytical performance, including limit of blank (LoB) and limit of detection (LoD), was evaluated using cell line DNA, and clinical cut-offs were established using receiver operating characteristic analysis. Results: The ddPCR assay demonstrated high analytical sensitivity, with LoD values of 0.075% for BAT-26, 0.1% for ACVR2A, and 0.025% for DEFB105A/B. Using optimized clinical cut-offs, the concordance rate between ddPCR and conventional PCR assays was 98.4% in tissue samples. Marker performance varied by cancer type, with reduced sensitivity observed in endometrial cancer. In plasma samples, MSI-H was detected in 1 of 21 cases (4.8%), and the overall concordance rate with tissue-based MSI status was 94.7%. Conclusions: The ddPCR assay demonstrated high concordance with conventional MSI testing methods and showed potential as a sensitive tool for MSI detection in both tissue and plasma samples. However, optimization of marker panels and establishment of sample-type-specific clinical cut-offs are required, particularly for ctDNA-based analysis. Further large-scale studies are needed to validate the clinical utility of ddPCR for MSI detection and monitoring. Full article

Background/Objectives: The increasing use of pea protein in plant-based foods raises concerns about IgE-mediated reactions, particularly in individuals sensitized to peanut. Knowledge on clinically relevant pea allergens and the impact of heat processing remains limited. This study investigated how thermal treatment affects the IgE binding and functional allergenicity of pea proteins in children with a confirmed pea allergy, with or without a concomitant peanut allergy. Methods: Serum from 11 patients was analyzed using SDS-PAGE, Western blotting, and an indirect basophil activation test (iBAT). Results: All patients showed IgE binding to Pis s 1 and PA2a/b in raw pea extract, with variable sensitization to Pis s 2 and mitogenic lectin. Heating (120 °C, 5 min) markedly reduced IgE binding and eliminated detectable IgE to Legumin S and ML. Despite this reduction, basophil sensitivity did not decrease; in several patients, EC50 values significantly decreased, indicating increased basophil responsiveness to heated pea. Patients with IgE profiles dominated by Pis s 1 and PA2a/b were most likely to show enhanced basophil activation after heating. Conclusions: These findings demonstrate that heat-stable vicilin subunits and albumins can maintain functional allergenicity despite reduced IgE recognition, underscoring the need for diagnostic approaches that incorporate processed food allergens. Full article

Exercise training improves maximum aerobic capacity, in part, through improvements in skeletal muscle function. This study aimed to investigate adaptations to improved aerobic capacity training through non-invasive and non-exhaustive tests of hyperemic muscle blood flow and near-infrared spectroscopy (NIRS) muscle oxygenation kinetics. An experimental study was conducted on 18 participants (age, 28.2 ± 5.3 yr; _absVO_2max, 3.60 ± 0.67 L·min⁻¹). Before and after the intervention of a 6-week of high-intensity interval training (HIIT), participants underwent three tests: (1) a graded cardiopulmonary exercise test; (2) a vascular occlusion test; and (3) a steady-state exercise (SSE) at 60% of PPO. Expired gas analysis, superficial femoral blood flow (occlusion test only) and SmO₂ kinetics were measured. The intervention increased maximal aerobic capacity _absVO_2max (p < 0.001, d = 0.65) and PPO (p < 0.001; d = 0.41). Moreover, steady-state _absVO₂ (p = 0.006; d = 0.37) and HR (p = 0.001; d = 0.65) were reduced. With the cuff test, the SmO₂ desaturation slope increased (p = 0.04; d = 0.52), while peak muscle blood flow (p = 0.02; d = 0.51) and the SmO₂ 10 s reoxygenation rate increased (p < 0.001 d = 1.11; 0.74 ± 0.28 to 1.17 ± 0.45%/s). During steady-state exercise, SmO₂ decreased less (p = 0.02; d = 0.43), and the 10s recovery kinetics rate was slowed (p = 0.01 d = 0.30; 0.28 ± 0.20 to 0.22 ± 0.21%/s). The improvement in VO_2max had a moderate correlation with the SmO₂ recovery rate post-steady-state exercise (p = 0.05, r = −0.54). HIIT changed maximal aerobic capacity alongside improvements in skeletal muscle hyperemic blood flow, SmO₂ post-occlusive reactive hyperemia and SmO₂ post-exercise recovery kinetics. Thus, the findings indicated that non-invasive and non-exhaustive hemodynamic kinetic profiles can monitor adaptations to improved aerobic capacity. Full article

Controlled-release fertilizer (CRF) improves fertilizer-use efficiency through sustained nutrient release, but its rate-dependent effects on the growth and physiology of Paeonia delavayi seedlings remain unclear. In this study, germinated seeds of P. delavayi with radicles 3–4 cm in length were grown under container nursery conditions with four CRF application rates: (CK, 0 kg·m⁻³), treatment 1 (T1, 0.6 kg·m⁻³), treatment 2 (T2, 1.2 kg·m⁻³), and treatment 3 (T3, 2.4 kg·m⁻³). Morphological traits, root characteristics, biomass accumulation, physiological parameters, and chlorophyll fluorescence were evaluated, and Pearson correlation and fuzzy membership analyses were used to compare overall treatment performance within the tested range. CRF significantly promoted seedling height, leaf number, petiole length, and biomass accumulation, although the promoting effect did not increase continuously with fertilizer rate. By June, seedling height in T2 was 160% greater than that in CK, while aboveground biomass increased by 552% and 574% in T2 and T3, respectively. Root morphological traits were not significantly affected, suggesting that CRF primarily promoted aboveground development and biomass production. Medium and high CRF rates increased leaf superoxide dismutase (SOD) activity by 42% and 103%, respectively, and peroxidase (POD) activity by 163% and 250%, respectively. Aboveground starch content was 45% higher in T2 than in CK. In contrast, photosynthetic pigment contents and the chlorophyll a/b ratio were not significantly affected by CRF. Chlorophyll fluorescence analysis showed that Fv/Fm remained stable among CRF treatments (0.78–0.82) and was significantly higher than that in CK (0.65), whereas the actual quantum yield of PSII [Y(II)] did not differ significantly among treatments. Relative to CK, the quantum yield of non-photochemical quenching [Y(NPQ)] increased from 0.20 to 0.40 in T2, while the quantum yield of non-regulated energy dissipation in PSII [Y(NO)] decreased from 0.37 to 0.24–0.22 in T2–T3. Pearson correlation and fuzzy membership analyses ranked the treatments as T2 > T3 > T1 > CK, indicating that T2 performed most favorably within the tested range, although its advantage over T3 was small. Overall, an appropriate CRF rate promoted P. delavayi seedling growth and was associated with changes in biomass accumulation, antioxidant enzyme activity, carbon assimilate storage, and chlorophyll fluorescence parameters. Full article

A realistic description of halo nuclei, characterized by low-lying breakup thresholds, requires a proper treatment of continuum effects. We have developed an ab initio approach, the No-Core Shell Model with Continuum (NCSMC), capable of describing both bound and unbound states in light nuclei in a unified way. With chiral two- and three-nucleon interactions as the only input, we can predict the structure and dynamics of halo and other light nuclei and, by comparing to available experimental data, test the quality of chiral nuclear forces. We review NCSMC calculations of weakly bound states and resonances of the exotic halo nuclei ${}^6$He, ${}^8$B, ${}^{11}$Be, and ${}^{15}$C. For the latter, we discuss its production in the capture reaction ${}^{14}$C(n,$\gamma$)${}^{15}$C. We highlight the challenges of a description of ${}^6$He as a Borromean n-n-${}^4$He system. Finally, we present our calculations of excited states in ${}^{10}$Be exhibiting a one-neutron halo structure and a large scale No-Core Shell Model investigation of ${}^{11}$Li as a precursor of a full n-n-${}^9$Li NCSMC study. Full article

Chronic wounds represent a major complication of underlying conditions such as diabetes mellitus, arterial ischemia, surgical wound and burns. This study aimed at the phenotypic and molecular characterization of antimicrobial resistance for a selection of bacterial isolates, originating from wounds harvested from patients hospitalized in the Vascular Surgery and Plastic Surgery wards. The microbiological diagnosis of wound infections was established according to the laboratory’s working protocol. PCR screening of antibiotic resistance genes was performed using a real-time PCR, while the microtiter plate assay was used to determine the biofilm-forming capacity. Testing of biofilm susceptibility to meropenem and amikacin was performed on Calgary biofilm device. Of the 88 bacterial isolates studied, 78.40% were Gram-negative bacilli (GNB)—Klebsiella pneumoniae (K.P), Pseudomonas aeruginosa (P.A), Proteus mirabilis (P.M), Acinetobacter baumannii (A.B), while the remaining 21.60% were Gram-positive cocci (GPC)—Staphylococcus aureus (S.A). All A.B isolates and 92.59% of K.P were carriers of β-lactamase- and carbapenemase-encoding genes, while 57.89% of S. aureus isolates were carriers of mecA (methicillin-resistant). Strong biofilm-forming isolates (B+++) were more frequent in P.A than in K.P (p = 0.002) and P.M (p = 0.02), with a frequency comparable to that of A.B strains (p = 0.212). When analyzing the biofilm reaction to meropenem, a significantly lower susceptibility was detected in the biofilm for K.P isolates, compared to the planktonic ones. Most GNB have been extensively multidrug-resistant, particularly K.P and A.B. Isolates from chronic wounds are major biofilm-formers. A strong and statistically significant association has been identified in the case of K.P and P.M between the presence of resistance genes and the biofilm-forming capacity. These findings highlight the need for a customized therapeutic approach for each chronic wound, considering the mechanisms underlying treatment resistance. These include bacterial virulence factors and the wound microenvironment colonized by the biofilm and the relative contribution of each to the overall resistance profile. Full article

The construction sector’s drive for sustainability has increased the use of Cross-Laminated Timber (CLT), yet its structural reliability is governed by the integrity of the adhesive bond line. This study evaluates the influence of three one-component polyurethane (PUR) formulations (R1, R2, R3) on the adhesion performance of maritime pine CLT. To isolate adhesive-related effects, lamellas were mechanically classified by modulus of elasticity (MOE) and randomly allocated within stiffness classes. Adhesive characterization through ABES, FTIR, and DSC revealed that R3 exhibited slower cure kinetics (t₀ = 5482 s) but higher thermal stability. Mechanical testing showed that all formulations developed structurally effective dry bonds with shear strengths exceeding 7.1 MPa, with R3 achieving significantly higher dry shear and interlaminar strength. However, 24 h water immersion caused a catastrophic strength reduction exceeding 95% across all formulations, shifting the failure mode from the wood substrate to the adhesive layer. DSC analysis identified glass transition temperatures between 28 °C and 32 °C, which are consistent with the potential for moisture-induced plasticization near service temperatures. These results indicate that while slower-curing formulations like R3 enhance bond quality in dense softwoods due to improved interphase formation, all evaluated PUR systems showed significant vulnerability to saturated conditions, suggesting that adequate moisture protection is essential for maritime pine CLT applications. Full article

Antibiotic resistance (ABR) is a significant threat to human and animal health, as well as to the environment. Antibiotic residues, antibiotic-resistant bacteria and antibiotic resistance genes are emerging contaminants of water resources. The present study aimed to assess the prevalence of ABR among waterborne enterococci in an anthropogenic-affected section of the Yantra River (Bulgaria). The susceptibility of 426 strains to 13 antibiotics (ABs) was tested by the disk diffusion method, and the genes encoding resistance by PCR analyses. A total of 39% of isolates were found to be antibiotic-resistant, with 9% mainly being multidrug-resistant to three AB classes. The most common resistance was to erythromycin (19%), tetracycline (18%) and ampicillin (14%), encoded by the ermB, tetM and blaTEM genes. A total of 3% of isolates were ciprofloxacin-resistant and only 1% was resistant to vancomycin or high-level gentamicin. All isolates were susceptible to teicoplanin and linezolid. Spatial variations in ABR levels were found, with the lowest abundance of antibiotic-resistant enterococci occurring in upstream river waters, away from urban areas, and the highest in urban areas. The spread of waterborne antibiotic-resistant enterococci highlights the need for water pollution management, monitoring and control to limit anthropogenic pressures through wastewater discharges and diffuse fecal pollution, and to ensure the ecological well-being of receiving waters. Full article

To address issues such as thermal stress concentration in metal bone implants produced via high-energy beam direct additive manufacturing, a method was proposed to fabricate porous titanium scaffolds. This approach combined Fused Deposition Modeling (FDM) with a debinding–sintering process. Ti/ABS composite filaments with titanium volume fractions of 35%, 40%, and 45% were successfully developed via a single-screw extrusion process. Their feasibility in the FDM process was subsequently verified. The effects of different processing parameters on the forming quality and dimensional accuracy of the green bodies were investigated. After debinding and sintering the composite scaffolds prepared with optimized parameters, structurally intact porous titanium scaffolds were obtained. Microscopic characterization shows that the scaffold surface consists primarily of titanium, and the pore structure remains intact. Furthermore, compression tests were performed on three types of porous titanium scaffolds with different porosities. The results indicate that the combination of ABS/titanium alloy composite filaments, FDM technology, and debinding–sintering post-processing enables the high-quality and efficient production of porous titanium scaffolds. The elastic modulus of the resulting scaffolds ranges from 1.2 to 1.6 GPa, and the compressive strength is between 25.7 and 68.3 MPa. The elastic modulus matches that of human cancellous bone. Meanwhile, the compressive strength is significantly higher than that of cancellous bone and falls between the values for cancellous and cortical bone. These mechanical properties meet the requirements for human bone, providing a new approach for the manufacture of orthopedic implants. Full article

Bst DNA polymerase is a biotechnologically modified thermostable enzyme from the thermophilic Gram-positive bacterium Geobacillus stearothermophilus. The unique structure of Bst DNA polymerase determines its thermal stability, ability to replace a DNA strand and specificity. The high specificity of Bst DNA polymerase ensures the efficiency, sensitivity, and high rate of loop-mediated isothermal amplification (LAMP), which is widely used in vitro biotechnology. The review reveals the structural and functional features of the enzyme, its application in LAMP and methods of improvement of thermal stability (including directed evolution, site-directed mutagenesis, fusion constructs, and chemical modifications). The terminal transferase activity and ab initio synthesis are discussed regarding problems of Bst DNA polymerase and the ways to eliminate them. The questions of introducing modified nucleotides and primers to expand the diagnostic capabilities of LAMP are also discussed. Modern advances in Bst DNA polymerase engineering pave the way for the creation of reliable, thermostable, and highly specific test systems suitable for widespread diagnostic applications. Full article

This study evaluated the effects of abiotic factors and extraction conditions on the yield, chemical composition, and antimicrobial activity of essential oil (EO) from Pistacia lentiscus L. leaves collected at four Adriatic locations during three phenological stages. Steam distillation was performed at 0.3, 0.7, and 1 bar. EO yield increased significantly with pressure, reaching a maximum at 1 bar, while the flowering stage provided the highest yields overall. Leaves from Vela Luka produced the highest EO yield, whereas Pag samples yielded the least. GC–MS analysis identified 56 components, accounting for 99.19–99.99% of total EO, with α-pinene, limonene, myrcene, and β-pinene as the dominant constituents, confirming a monoterpene-rich chemotype. All EO samples showed low but measurable inhibitory activity against Escherichia coli AB1157 and Erwinia amylovora EaED, as assessed by the disk diffusion method. Pearson correlation and PCA analyses indicated a positive association between monoterpene content and inhibition zone diameter against E. coli, and a positive association between monoterpene alcohol content and inhibition against E. amylovora. As antimicrobial activity was assessed exclusively by the disk diffusion method, the present findings may serve as an indicative basis for future investigations into the relationship between EO chemical composition and antimicrobial potential, and they require validation through quantitative, standardized antimicrobial testing. Full article

Background: The increasing use of multigene germline testing in breast cancer has expanded the detection of pathogenic/likely pathogenic (P/LP) variants and variants of uncertain significance (VUSs) across a broad range of breast cancer susceptibility genes. However, the extent to which pathogenic certainty and penetrance level are associated with distinct clinicopathological phenotypes and management patterns remains incompletely defined. Methods: In this multicenter retrospective study, we included 405 breast cancer patients with germline alterations in breast cancer susceptibility genes. Patients were classified into three groups: high-penetrance P/LP (n = 116), moderate/low-penetrance P/LP (n = 69), and VUS (n = 220). The primary endpoint was non-luminal invasive breast cancer phenotype, defined as HER2-positive or triple-negative disease versus luminal A/B disease. Multivariable logistic regression was performed to assess the association between genetic group and non-luminal phenotype, adjusted for age at diagnosis and family history. Secondary analyses evaluated management patterns across groups and examined a prespecified non-BRCA cohort. Results: Patients with high-penetrance P/LP variants were younger at diagnosis and had higher Ki-67 values than those in the other groups. Molecular subtype distribution differed significantly across groups, with triple-negative disease most frequent in the high-penetrance P/LP group (33.9%) compared with the moderate/low-penetrance P/LP (9.8%) and VUS (12.9%) groups. In multivariable analysis, high-penetrance P/LP status was independently associated with increased odds of non-luminal phenotype compared with VUS (OR 1.79, 95% CI 1.06–3.04; p = 0.029), whereas moderate/low-penetrance P/LP status was not (OR 0.96, 95% CI 0.48–1.91; p = 0.911). Management patterns also differed significantly. Initial mastectomy was performed in 65.4% of evaluable patients in the high-penetrance P/LP group, compared with 46.4% in the moderate/low-penetrance P/LP group and 45.5% in the VUS group. Final bilateral mastectomy was observed in 62.6%, 17.5%, and 9.9%, respectively (p < 0.001). In the prespecified non-BRCA analysis, non-BRCA P/LP status was not associated with non-luminal phenotype compared with non-BRCA VUS (OR 0.83, 95% CI 0.40–1.72; p = 0.609). Conclusions: Among breast cancer patients with germline alterations in breast cancer susceptibility genes, high-penetrance P/LP variants are associated with a distinct clinicopathological profile characterized by younger age at diagnosis, higher proliferative activity, and increased likelihood of non-luminal disease. In contrast, moderate/low-penetrance P/LP variants, especially VUSs, do not show the same degree of phenotype specificity. These findings support a framework in which pathogenic certainty and penetrance level remain central to the interpretation of multigene germline testing in breast cancer. Full article

Ensuring the safety of sustainably managed medicinal plants is closely linked to the quality of plant raw materials, including the presence of heavy metals within safe limits. Sustainable management in the context of herbal raw materials therefore entails responsible management of herbal plant resources, integrating environmental protection with ensuring long-term economic profitability. The aim of this study was to analyze selected biochemical parameters and to determine metal concentrations in soils and leaves of Plantago lanceolata L. collected from natural habitats at increasing distances from traffic routes. The content of Zn, Cu, Ni, and Pb was determined in the soils and leaves of Plantago lanceolata L. Assessing the content of these elements in plant raw materials allows for: the prevention of harmful substances in final products, adaptation of raw materials to applicable safety standards (avoiding toxicity), and protection of consumer health. This promotes sustainable development by building a safe supply chain. The leaves of Plantago lanceolata L. were also tested for biochemical enzymatic (catalase (CAT) and superoxide dismutase (SOD)) and non-enzymatic (chlorophyll a and b (Chl a and b), carotenoids (Car), ascorbic acid (AAC)), and mechanisms regulating the activity of reactive oxygen species (ROS) were determined in the leaves of Plantago lanceolata L. Based on the results of leaf pH, relative water content (RWC), ascorbic acid content, and total chlorophyll content, the air pollution tolerance index (APTI) was calculated. The distance from the road has a significant impact on the concentration of the heavy metals analyzed. The soils were found to be free of Zn, Cu, Pb, and Ni contamination. However, analysis of Plantago lanceolata L. leaves revealed exceedances of acceptable lead limits for herbal plants. The content of pigments, the ratio of Chl a/b, and Chl (a + b)/Car in the leaves of Plantago lanceolata L. was significantly dependent on the distance from the road. The activity of CAT and SOD in the leaves of Plantago lanceolata L. growing closest to the road was significantly higher compared to the others. APTI values suggest that Plantago lanceolata L. exhibits sensitivity to pollution, independent of its distance from the emission source. Full article

Despite its advantages, 3D printing may expose users to volatile organic compounds (VOCs) and particle emissions. Emissions from commercially available acrylonitrile-butadiene-styrene (ABS)- and polyethylene terephthalate glycol (PET-G)-based filaments were analyzed to evaluate differences among material formulations from multiple manufacturers. Chamber-based measurements and complementary thermal decomposition experiments were used to characterize particle number concentrations and chemical emissions. The highest particle emissions occurred during the initial warm-up and the final stages of the printing process. The ABS-based filaments tested in this study exhibited higher VOC emissions, dominated by styrene (up to 264.75 μg/m³), and particle number concentrations approximately one order of magnitude greater than those measured for the tested PET-G-based filaments. The dominant particle sizes ranged from 55 to 90 nm. PET-G-based filaments showed higher thermal stability but emitted notable concentrations of acetaldehyde (up to 70.93 μg/m³) and phthalic acid esters. Both filament types released compounds of potential health concern, including formaldehyde and reprotoxic substances such as dibutyl phthalate and bis(2-ethylhexyl) phthalate. Differences were observed among fibers made from the same polymer type, indicating the influence of formulation-specific factors. These results underscore the importance of material selection and adequate ventilation to minimize exposure during 3D printing. Full article

Background: The integration of active breaks during the school day has been widely demonstrated to be effective in counteracting sedentary behaviors. The present study assessed the efficacy of a structured active breaks (ABs) intervention implemented during recess on multiple domains of Physical Literacy (PL) in primary-school children. Methods: A single-blind randomized controlled trial was conducted with 139 children (aged 9–10 years). Classes were randomized into an Experimental Group (EG, n = 66) and a Control Group (CG, n = 73). The EG participated in an 8-week intervention (six sessions/week, ~10 min) consisting of coordinative and interdisciplinary motor tasks during recess. Pre- and post-intervention assessments included physical fitness (SLJ, 4 × 10 m SR, 6MWT, MBT), gross motor skills (TGMD-2), selective attention (Bell Test), physical activity levels (PAQ-C), physical self-perception (PSP), and enjoyment (PACES). Results: A mixed-design MANOVA revealed a significant multivariate Time × Group interaction (p < 0.001). Univariate analyses showed significant improvements in the EG compared to the CG for explosive strength (p < 0.001), agility (p < 0.001), Gross Motor Quotient (p = 0.003), and selective attention (p < 0.001). Furthermore, the EG demonstrated significant increases in physical activity levels, self-perception, and enjoyment (p < 0.05). No significant gender interaction was found, indicating equal effectiveness for boys and girls. Conclusions: Transforming recess into a structured opportunity for movement through ABs effectively enhances physical, cognitive, and affective domains. This intervention represents a sustainable strategy for Health-Promoting Schools to foster PL and psychophysical well-being without reducing curricular instruction time. Full article