Search Results (502)

Accurate quantification of singlet oxygen (¹O₂) production is central to photodynamic therapy (PDT) dosimetry, yet direct measurement remains challenging due to the weak and transient nature of ¹O₂ luminescence. To our knowledge, this study presents the first controlled exploratory cross-platform comparison of Multispectral Singlet Oxygen Luminescence Dosimetry (MSOLD) and Time-Resolved Singlet Oxygen Luminescence Detection (TSOLD) under matched irradiation conditions. MSOLD enables continuous monitoring during irradiation through spectral decomposition of near-infrared emission, whereas TSOLD uses pulsed excitation and time-correlated single-photon counting to provide lifetime-resolved ¹O₂ measurements. Across PpIX concentrations of 10–100 mg/kg, both methods detected irradiation-induced decreases in ¹O₂ luminescence, while TSOLD provided stable lifetime estimates of approximately 10 µs in the methanol-rich phantom. Across fluence rates of 1–5 W/cm², both techniques showed progressively larger irradiation-induced reductions in ¹O₂ luminescence with increasing irradiance. Cumulative MSOLD-derived ¹O₂ signals showed an approximately linear relationship with Singlet Oxygen Explicit Dosimetry (SOED) calculations (R² > 0.98) under the investigated conditions. Together, these results demonstrate consistent concentration- and fluence-rate-dependent trends between MSOLD and TSOLD and highlight their complementary measurement characteristics: MSOLD provides continuous spectral monitoring during irradiation, whereas TSOLD provides lifetime-resolved characterization of ¹O₂ and photosensitizer kinetics. These findings provide a basis for future replicated and biologically relevant studies of luminescence-based PDT dosimetry. Full article

Background: Pooling public blood transcriptome studies never designed for integration introduces failures that standard diagnostics miss. Objective: To define safeguards for confounding-aware integration and establish what an integrated stroke dataset can support. Methods: Eleven GEO series (1885 samples) were integrated under six safeguards: expression fingerprinting for undeclared patient overlap; staged ComBat with leave-one-study-out PCA; scale-correct deconvolution; composition-adjusted differential expression; an estimability audit with leakage-safe cross-validation; and a provenance audit against the source studies’ deposited tables. Results: Fingerprinting found 33 shared patients in two series. After ComBat, study and platform explained ~0% of PC1, and cell composition explained 61.4%. Linear rather than log-scale deconvolution input changed the neutrophil fraction (0.07 to 0.25) but did not affect the level of cross-method agreement (r = 0.81 vs. 0.78). Neutrophil adjustment removed 65–84% of acute differentially expressed genes, and co-expression modules re-correlated with composition even among composition-robust genes. Stroke subtype was fully confounded with study (classifier AUC 0.882 from single-class folds). The provenance audit reclassified our novel candidate finding in cell-sorted data as replicative and confirmed the depositors’ differential expression (monocytes: r = 0.92, neutrophils: 0.90). The integrated dataset supported a direction-resolved acute signature, three temporal response programs in a 513-gene core, and seven candidate stroke-versus-trauma discriminators with divergent kinetics. Conclusion: Passing integration diagnostics does not guarantee estimability. Estimability must be assessed before correction removes the evidence, and novelty must be checked against deposited data. Full article

Fresh black soldier fly larvae (BSFL) have a high moisture content that complicates storage and requires energy-intensive postharvest drying. This study evaluated the effects of drying-air temperature and tray rotation speed on BSFL drying kinetics, energy performance, and moisture transport in a rotating-tray hot-air dryer. A 3 × 3 factorial experiment was conducted at 50, 60, and 70 °C and tray rotation speeds of 0, 5, and 10 rpm, with three independent replicates, while drying-air velocity was maintained at 3.0 m s⁻¹. Drying time decreased from 31.0 h at 50 °C and 0 rpm to 6.5 h at 70 °C and 10 rpm. The latter condition achieved the highest drying efficiency (49.11%) and lowest specific energy consumption (4.60 MJ kg⁻¹ water removed). Comparable values were obtained at 60 °C and 10 rpm (49.07% and 4.61 MJ kg⁻¹ water removed, respectively). Among nine thin-layer models evaluated for the six rotating-tray treatments at 5 and 10 rpm, the Midilli model provided the best fit. Apparent effective moisture diffusivity ($D_{eff})$ increased from 0.2457 × 10⁻⁹ to 1.2946 × 10⁻⁹ m² s⁻¹, while apparent activation energy ranged from 67.39 to 68.46 kJ mol⁻¹. Rotating-tray drying accelerated BSFL dehydration and improved engineering performance; however, microbial safety and product quality require separate evaluation before food or feed recommendations can be made. Full article

In this study, we evaluated the replication kinetics and protective efficacy of the gene-deleted Marek’s disease (MD) vaccine strain SC9-1 in specific-pathogen-free (SPF) chickens immunized with different doses. Chickens were divided into two groups: one group received a high dose (5000 PFU) and the other a low dose (500 PFU) of the vaccine at 1 day of age, with a non-vaccinated group serving as the control. Vaccine viral copy numbers in the spleen and feather tips were detected by quantitative PCR (qPCR) at different days post-vaccination (dpv) (3, 5, 7, 11, and 14 dpv). Meanwhile, a challenge trial was conducted where chickens were challenged with the very virulent Md5 strain (1000 PFU/bird) at 5 dpv and observed until 70 days of age. Results from the vaccination-only experiment revealed that viral replication in the spleen was detectable in 100% of chickens as early as 3 dpv, reaching its peak at 5 dpv, with the high-dose group exhibiting significantly higher viral loads than the low-dose group. In feather tips, replication was detectable in 100% of chickens by 5 dpv and peaked at 7 dpv, but no significant difference was observed between the two dose groups. The unvaccinated control group exhibited 100% mortality with characteristic gross and histopathological lesions of Marek’s disease. In stark contrast, both vaccinated groups (high and low dose) demonstrated 100% survival until the end of the observation period (70 days). No clinical signs or pathological abnormalities were observed in these groups. Furthermore, vaccination with SC9-1 did not adversely impact growth performance and the development of major immune organs, even following subsequent Md5 challenge. The results indicated that a higher initial vaccination dose of the SC9-1 vaccine was associated with elevated peak early in vivo replication level, whereas vaccination dose may not govern viral maturation and shedding levels of SC9-1 in feather-tip tissues. However, peak replication level was not a direct determinant of final protective efficacy. Notably, even the lowest tested dose (500 PFU) afforded full protection against early challenge with a very virulent MDV strain. Collectively, these observations contribute to our understanding of dose-dependent protection induced by gene-deleted MD vaccines and inform rational vaccination strategies. Full article

Environmentally sustainable feed additives that mitigate enteric methane (CH₄) emissions are a priority for climate-smart livestock production. The present study investigated the effects of laurel (Laurus nobilis) ethanolic extract applied as a normal-sized bulk extract (B) or an oil-in-water nanoemulsion (N) at inclusion levels of 0, 15, 30, and 45 µL/g dry matter (DM) on in vitro gas production (GP) kinetics, CH₄ and carbon dioxide (CO₂) emissions, nutrient degradability, and rumen fermentation end-products. A 2 × 3 + 1 factorial arrangement within a completely randomized design yielded seven treatment groups: Control, B15, B30, B45, N15, N30, and N45, with three incubation runs serving as replicates. The asymptotic GP (b) and gas yield at 24 h (GY₂₄) exhibited linear and quadratic responses within both laurel extract forms, peaking at 15 µL/g DM for bulk (b: 332.9 mL/g DM; GY₂₄: 377.8 mL/g DM) and nano (b: 342.2 mL/g DM; GY₂₄: 385.5 mL/g DM) treatments before declining at higher doses, with N15 producing the highest values overall (p < 0.05). Metabolizable energy followed the same linear and quadratic trends (p < 0.001), reaching 9.01 MJ/kg DM (B15) and 9.40 MJ/kg DM (N15) versus 8.01 MJ/kg DM in the control. Both laurel extract forms linearly (p < 0.001) reduced CH₄ as a percentage of total gas; however, on an absolute mL/g DM basis, CH₄ output declined only at 30 and 45 µL/g DM, whereas B15 and N15 showed a slightly higher absolute CH₄ output than the control alongside greater total gas production; N45 achieved the greatest absolute reduction (45.9 mL/g DM versus 75.6 mL/g DM in the control, representing a 39.3% reduction). The nanoemulsion form was consistently superior to the bulk form in reducing CH₄ concentration (p < 0.001). Dry matter degradability peaked at N15 (61.0% versus 57.5% in the control) and declined at higher nano doses to 49.2% at N45 (p < 0.001). Total volatile fatty acid concentrations and individual acetate, propionate, and butyrate values were highest at 15 µL/g DM for the nanoemulsion and declined with increasing dose (p < 0.01). Rumen pH increased linearly (p < 0.01), and ammonia-N decreased as dose escalated (p < 0.001). The superior fermentation response of the nanoemulsion at 15 µL/g DM, together with the greater and dose-dependent CH₄ reduction achieved at 30–45 µL/g DM, underscores the capacity of nanoemulsification to enhance the bioavailability and dose-efficiency of laurel extract bioactives in the rumen, while indicating that no single dose tested here simultaneously maximized fermentation and CH₄ mitigation. These in vitro findings support laurel extract nanoemulsion as a promising environmentally sustainable feed additive candidate for enteric CH₄ mitigation, warranting an in vivo dose–response evaluation to identify a practical inclusion level that balances CH₄ mitigation, fermentation efficiency, and animal acceptability. Full article

Background: Transforming growth factor beta 1 (TGF-β1) is a master regulator of fibrogenesis and extracellular matrix (ECM) remodeling. Among molecules involved in profibrotic signaling, periostin (POSTN) and the Snail family transcriptional repressor 2 (SLUG, SNAI2) have emerged as potential regulators of tissue remodeling and fibroblast activation. However, the temporal relationship between TGF-β1-induced expression of SLUG and periostin in human dermal fibroblasts remains poorly understood. Objectives: This study aimed to evaluate the effect of TGF-β1 stimulation on SNAI2/SLUG and POSTN/periostin expression dynamics in normal human dermal fibroblasts (NHDFs) and characterize their association with profibrotic responses. Methods: NHDFs were stimulated with recombinant human TGF-β1 (20 ng/mL) for 24, 48, and 72 h. SNAI2 and POSTN mRNA expression was quantified by RT-qPCR, intracellular SLUG protein levels were assessed by Western blotting, and periostin accumulation in the culture medium was measured by enzyme-linked immunosorbent assay (ELISA). The RT-qPCR and ELISA data were analyzed using two-way ANOVA with Bonferroni correction; because of the small number of replicates (n = 2), the Western blot data are presented descriptively. Results: TGF-β1 significantly increased POSTN mRNA expression after 24 and 48 h (both p < 0.001); at 72 h, POSTN mRNA levels remained numerically elevated relative to the control, although this difference did not reach statistical significance. Periostin accumulation in the culture medium increased progressively and was significantly higher than in the control group after 48 and 72 h (p < 0.001). In contrast, SNAI2/SLUG exhibited transient expression kinetics, with increased SNAI2 mRNA at 48 h (p < 0.05) and the highest SLUG protein levels also observed after 48 h of stimulation, followed by a decline at 72 h. Conclusions: TGF-β1 induced time-dependent changes in SNAI2/SLUG and POSTN/periostin expression in human dermal fibroblasts. As fibroblast activation and ECM remodeling were not directly assessed, the present conclusions are restricted to the expression changes in the two investigated molecules, whose concurrent yet temporally distinct regulation may indicate potential relevance in the fibroblast response to profibrotic signaling. Further studies are required to determine whether they participate in common regulatory mechanisms or independently contribute to fibrotic processes. Full article

Phosphate availability fluctuates widely in aquatic environments, yet how engineering the regulatory context of polyphosphate kinase affects cyanobacterial responses to such fluctuations remains unclear. Here, the wild-type strain of Synechococcus sp. PCC 7002 and Dppk, a modified ppk-expression strain derived from the WT background, were subjected to phosphate starvation followed by phosphate resupply. Dense RT-qPCR sampling was used to resolve rapid ppk-expression dynamics, and time-resolved RNA sequencing was performed at seven stages with four biological replicates per condition. WT displayed rapid, stage-dependent changes in ppk expression, including a pronounced late induction after phosphate resupply, whereas Dppk showed a more moderate temporal pattern. Transcriptome-wide analyses further revealed distinct basal states and response trajectories between the strains. Functional analyses linked these differences to photosynthesis-related processes, phosphate transport, and polyphosphate metabolism. These findings indicate that refactoring ppk expression alters both target-gene kinetics and the temporal organization of the broader phosphate-responsive transcriptional network. Full article

Monitoring bacterial contamination on surfaces is critical for hygiene and safety assurance in food, healthcare, and pharmaceutical settings, although routine methods still remain slow and laboratory dependent. Here, we report a portable respirometric platform based on sealed sensor sachets incorporating optical oxygen sensors to rapidly detect and quantify total aerobic viable counts (TVC) from swabbed surfaces. Following standardized surface swabbing, samples were incubated in the sachets and oxygen depletion kinetics were recorded with a handheld reader and microbial activity was inferred from oxygen consumption. Reference quantification was obtained by serial dilution and aerobic plate counting, enabling direct benchmarking of the respirometric readout against an industry-accepted culture method such as ISO 4833:2013. The platform demonstrated strong agreement with plate counts (R² > 0.94), achieving a median detection limit of 2.69 log₁₀ CFU/cm² and a dynamic range of 0–6 log₁₀ CFU/cm² for surface-associated microbial loads. The sensor system was also applied in a semi-industrial setting in a meat processing plant. Across replicate measurements, the assay provided consistent kinetic signatures and quantitative outputs, suitable for rapid and practical decision-making. Compared with traditional culture-based approaches (requiring up to 72 h), the respirometric sachets delivered actionable results within 10 h using a portable, low-infrastructure workflow, supporting rapid on-site hygiene verification and sanitation control. Full article

Transmissible gastroenteritis (TGE) and pseudorabies (PR) remain important viral diseases causing massive economic losses and posing a continuous burden on the global swine industry. The continuous emergence of variant strains of transmissible gastroenteritis virus (TGEV) and pseudorabies virus (PRV) has gradually weakened the protective efficacy of traditional vaccines, highlighting the urgent need for next-generation preventive vaccine candidates. Here, we constructed a recombinant pseudorabies virus named rPRV-AD expressing the major neutralizing epitopes A and D of the TGEV spike protein via homologous recombination combined with CRISPR/Cas9-gene editing technology and then evaluated its biological characteristics in vitro and immunogenicity in piglets. The results showed that this recombinant virus exhibited similar replication kinetics and biological properties to the parental strain. Immunization of 2-week-old piglets with rPRV-AD caused no obvious adverse effects and induced specific antibody and neutralizing antibody responses against both TGEV and PRV. Following virulent TGEV challenge, compared with the DMEM control group, rPRV-AD immunization alleviated clinical signs of piglets and significantly reduced viral load in intestine and feces, although its protective efficacy was lower than that of the commercial TGEV vaccine. Moreover, rPRV-AD provided effective clinical protection against challenge with the virulent PRV NY strain. In summary, these findings suggest that rPRV-AD represents a vaccine candidate that provides partial protection and warrants further optimization, and yet shows short-term protective efficacy against both PRV and TGEV in pigs. Full article

Corrosion-severity grading of grounding-grid connectors from optical images supports proactive power-infrastructure maintenance. Existing approaches rely on single-time-point, manually thresholded hue–saturation–value (HSV) metrics and static multi-criteria decision-making (MCDM) frameworks that cannot capture corrosion dynamics. In this paper we present a pipeline that (1) defines a four-class corrosion grade from an HSV area fraction (S_corr) measured on RGBA optical images, and validates those labels against a baseline-referenced CIEDE2000 metric zero-referenced to each connector’s as-received appearance; (2) generates 240 color-prior-constrained procedural synthetic images from 53 real images across six connector types; (3) fine-tunes a ResNet-18 to estimate corrosion coverage continuously, deriving the reported severity class from that estimate rather than predicting it directly; and (4) fits power-law kinetics C(t) = k·tⁿ to the S_corr time series, propagates bootstrap uncertainty into a Technique for Order Preference by Similarity to Ideal Solution (TOPSIS) framework, and reports kinetics-aware rankings as rank probabilities. The label validation quantifies two limitations of single-threshold HSV grading: a material-color offset that scores an unexposed copper connector at S_corr = 0.442, and insensitivity to achromatic corrosion products covering roughly 80% of the aluminum and galvanized-steel surface. Ablation experiments replicated over five random seeds show that neither contribution claimed from a single run survives replication: synthetic augmentation changes macro-F1 by +0.050 (p = 0.46) under the adopted checkpoint-selection rule and by −0.059 (p = 0.43) under the rule used in the original experiments, and the monotonicity-consistency loss by −0.011 (p = 0.87) and +0.001 (p = 0.99) respectively; the previously reported single-run values of 0.208 and 0.494 are draws from opposite tails of the same seed distributions (0.403 ± 0.140 and 0.344 ± 0.073). The one formulation that improves significantly is the continuous one adopted here, which raises Spearman agreement with the independent metric from 0.316 ± 0.150 to 0.698 ± 0.108 (p = 0.005). Measured against controls, a classifier that never sees the image reaches macro-F1 = 0.425 and, after Holm–Bonferroni correction, no deep configuration is distinguishable from it; none exceeds a one-dimensional linear rule on S_corr (0.664); and under leave-one-material-out cross-validation the network does not improve on S_corr used directly as a predictor (ρ = +0.627 against +0.744, paired p = 0.14). Time-resolved energy-dispersive X-ray spectroscopy (EDS) provides a partial chemical consistency check, with welding at ρ = 0.82 (raw p = 0.023), but no material survives Holm correction across the six tested. A U-Net segmentation head supervised only by synthesis-derived masks attains Dice = 0.85 in-domain and collapses to a 0.033 output range on real images, 5% of the HSV metric’s range; the photometric-stability advantage previously claimed for it is an artifact of that collapse and is withdrawn. Kinetics-aware MCDM with propagated uncertainty resolves 9 of 15 pairwise orderings, placing stainless steel above welding at 30 chamber days with probability 1.000 and reversing the static ranking. The pipeline, code and fixed data split are fully reproducible (random seed 42). Full article

Proliferating cell nuclear antigen (PCNA) is an essential sliding clamp that coordinates nearly all DNA-templated processes. It does so by recruiting a diverse array of factors to DNA through PCNA-interacting protein (PIP) motifs on PCNA-binding proteins. Chromatin assembly factor 1 (CAF-1) is a histone chaperone that deposits histones onto silent regions of the genome immediately following DNA replication. PCNA recruitment of CAF-1 to the replication fork is essential for nucleosome assembly and epigenetic inheritance. In yeast, CAF-1 recruitment to PCNA is assumed to be facilitated using one PIP motif. Here, we identify a second PIP motif in CAF-1, designated PIP1, located within the N-terminal intrinsically disordered region of the protein. Binding kinetics demonstrate that the PIP1 motif sequence binds PCNA with substantially lower affinity than the previously characterized PIP motif, designated PIP2. Structural and binding data reveal PIP1 as an extended PIP motif, in which C-terminal flanking residues make extensive, atypical contacts with PCNA that significantly increase its affinity. Neither PIP1 nor PIP2 alone is required for CAF-1–mediated gene silencing in vivo, but simultaneous disruption of both abolishes CAF-1 function. Together, these findings suggest CAF-1 engages PCNA multivalently and provide new insight into how PCNA selectively recognizes binding partners during nucleosome assembly. Full article

We analyse whether Darwinian natural selection acts as a metabolic-entropy-production-increasing process in a model of an open chemostat ecosystem, where Michaelis–Menten uptake kinetics are grounded in a thermodynamically consistent mesoscopic chemical-reaction-network model, and the continuum limit of the discrete replicator equation is derived as a Crow–Kimura reaction–diffusion process. On the quasi-static ecological manifold, an exact Fisher-type identity yields a monotonic increase in the metabolic entropy-production rate, proportional to the uptake-rate variance. Solving the reduced moment equations in closed form, under an explicit quasi-static and zero-skewness closure, identifies a thermodynamic ceiling imposed by mass conservation, while phenotypic trade-offs produce a finite sub-ceiling. When ecological and evolutionary timescales become comparable, a frozen-parameter Routh–Hurwitz analysis identifies an instantaneous spectral instability boundary whose scope and self-consistency are assessed for both unbounded and bounded trait models. The results of this analysis delineate the precise dynamical and biochemical conditions under which selection increases metabolic entropy production and the regimes in which that tendency is reshaped. Full article

Chagas disease control relies on chemical interventions, currently compromised by insecticide resistance. Triatoma virus (TrV) emerges as a biological candidate, requiring standardized methodologies to purify, quantify, and relate viral doses to biological effects in triatomines. We established a comprehensive protocol for the quantitative study of TrV using Rhodnius prolixus as a model, combining physical purification via sucrose gradients and absolute quantification by qPCR targeting the TrVgp1 gene to link viral load with host survival. Primers were validated, and a plasmid standard curve was constructed for absolute quantification. Four isolation methods from Triatoma infestans macerates were compared, including an optimized discontinuous gradient protocol (EGM). High (6 × ${10}^6$ viral genome copies/µL) and low (2.47 × ${10}^5$ viral genome copies/µL) doses of EGM-purified TrV were evaluated in oral infections of R. prolixus over 16 days post-inoculation (dpi). The EGM protocol achieved the highest yields, around ${10}^9$ virus genome copies/µL of purified viral stock. Intestinal viral replication was dose-dependent, exhibiting distinct kinetic phases between low (3–7 dpi) and high (9–12 dpi) inoculum cohorts. Only the high dose significantly reduced host survival. In conclusion, this protocol allows high-yield TrV purification and absolute quantification in R. prolixus, establishing a robust dose–response foundation to assess triatomine susceptibility for integrated biological control programs. Full article

Porcine epidemic diarrhea virus (PEDV) continues to cause devastating economic losses in the swine industry. However, the lack of species-specific in vitro models and reliance on costly animal models hinders development of effective interventions. We established a high-fidelity platform using porcine induced pluripotent stem cells (piPSCs; line VSMUi001-C) via a 28-day stepwise differentiation protocol. Successful lineage commitment was validated by OCT4 downregulation and robust FOXA2/SOX17 expression at day 5. By day 28, the piPSC-derived intestinal epithelial cells (piPSC-IECs) exhibited organized LGR5+ niches, MUC2+ goblet cells, and Villin+ enterocytes, mirroring native tissue architecture. Barrier functionality was confirmed by the continuous distribution of the tight junction protein zonula occludens-1 (ZO-1). Our results demonstrate that piPSC-IECs are permissive to PEDV and support productive infection, encompassing the complete viral life cycle from entry to release of infectious progeny. Productive replication was confirmed by direct immunodetection of viral antigen within inoculated cells. Viral challenge induced characteristic cytopathic effects and a specific pro-inflammatory elevation of tumor necrosis factor-alpha (TNF-α). Notably, infection fragmented ZO-1, providing a mechanistic explanation for the leaky gut phenotype observed in vivo. This platform serves as a robust system for investigating viral kinetics and host–pathogen interactions with high species-specific relevance. Full article

Humic acids (HAs) represent structurally heterogeneous supramolecular systems whose characterization by CP/MAS ¹³C NMR spectroscopy strongly depends on experimental conditions. Among these, contact time is one of the most important parameters controlling the efficiency of polarization transfer and the relative contribution of individual carbon groups. The aim of this study was to evaluate the influence of contact time on the apparent CP/MAS ¹³C NMR signal distribution of HAs isolated from different horizons of a permafrost peat soil (Hemic Folic Cryic Histosol) and to characterize their cross-polarization dynamics using kinetic modeling. A series of CP/MAS ¹³C NMR spectra was acquired at contact times ranging from 0.01 to 10 ms. Signal intensities of major structural groups were analyzed using both the classical three-parameter model and an extended four-parameter model of cross-polarization dynamics. The results demonstrated pronounced contact-time-dependent variations in the apparent CP/MAS signal distribution among alkyl, aryl, and carboxyl/amide structural domains. Short contact times preferentially enhanced proton-rich aliphatic and carbohydrate structures, whereas longer contact times increased the relative contribution of aromatic and carboxyl/amide fragments. The extended model provided a better fit for selected structurally heterogeneous spectral regions, particularly aromatic and heteroatom-containing domains, whereas the classical model adequately described the relatively homogeneous aliphatic regions. The fitted CP-dynamic parameters provided effective comparative descriptors of the investigated HAs under the experimental conditions employed and demonstrated consistent depth-dependent differences within the investigated peat profile. These depth-dependent differences may reflect changes associated with peat accumulation and long-term cryogenic preservation; however, this interpretation requires confirmation using independently replicated peat profiles. Full article