Search Results (11,665)

This study investigated a phototrophic approach to close nutrient loops by using landfill leachate as a culture medium to produce biomass and polyhydroxybutyrate (PHB) from a thermotolerant strain of Potamosiphon sp. A multi-cycle reuse scheme in which post-culture leachate was partially replenished with fresh leachate and reused in successive cultivation rounds to increase the biomass concentration (g/L) and the intracellular PHB content (% w/w) was tested. Three operational variables were optimized (leachate replenishment percentage, number of reuse cycles, and sanitation method (autoclaving, UV irradiation, or no treatment)) via the Box–Behnken response surface method. Both response variables were modeled with high predictive accuracy (R² = 0.98 for biomass and R² = 1.00 for PHB content). According to the experimental data, leachate replenishment emerged as the key factor influencing nutrient availability—particularly nitrogen and phosphorus—and thus PHB accumulation. The optimized conditions (2.17% v/v fresh leachate, three reuse cycles, and UV sanitation) yielded predicted values of 0.29 g/L biomass and 3.48% w/w PHB. These results demonstrate the feasibility of a controlled multicycle reuse process that integrates effluent treatment and biopolymer synthesis, offering a low-input, circular biotechnological approach for sustainable leachate valorization. Full article

The immune status of dogs is shaped by continuous exposure to antigenic and various environmental stimuli, which together influence the development, regulation, and effectiveness of immune responses. Stress-related immune alterations may not be evident at the systemic level but can emerge at cellular and molecular scales. Therefore, this study aimed to comprehensively characterize the hematological and immunological profiles of dogs in different environments. We evaluated lymphocyte responses under basal conditions and following CD3/CD28-mediated in vitro activation, with subsequent long-term culture. Gene expression analyses targeted markers of early T cell activation, cytotoxic effector function, cytokine signaling, and inhibitory immune regulation. The memory phenotype of T lymphocytes was evaluated after blood collection and prolonged in vitro culture. In addition, hematological and biochemical profiles were assessed, including basic parameters, cortisol, and C-reactive protein. Our results revealed that client-owned dogs exhibited lower baseline expression of activation markers, especially in comparison with the short-term stay group, indicating an early immune activation state upon entry to the shelter environment. Furthermore, T lymphocytes from short- and long-term shelter dogs exhibited marked differences in the distribution of naïve and effector-memory subsets as well as different expansion capacity. These alterations persisted during prolonged in vitro culture, indicating that stress duration and environmental antigen exposure differentially shape immune responsiveness. In summary, chronic stress modulates canine immune status in a time-dependent manner, highlighting the importance of integrated cellular and molecular approaches in assessing the impact of environmental stressors on dogs’ health and welfare. Full article

Chlamydia suis, a close relative of the human pathogen C. trachomatis, can be detected in the porcine gut, yet its prevalence and viability across intestinal segments remain poorly defined. This study aimed to assess the segment-specific prevalence, isolation success, and tetracycline susceptibility of C. suis in grower-finisher pigs. Jejunal, ileal, and colonic samples (n = 200 per intestinal segment) were collected from 600 pigs at slaughter and analyzed using C. suis-specific real-time PCR and culture. PCR revealed significantly higher detection rates in the colon (40%) than in the jejunum or ileum (both 4.5%), accompanied by significantly higher calculated bacterial loads in colonic samples. In contrast, viable C. suis was most frequently isolated from ileal material, indicating that the ileum may provide a more favorable condition for active bacterial replication. Among 24 culture-confirmed isolates, 75% were susceptible to tetracycline (MIC/MBC < 2 µg/mL), 12.5% exhibited an intermediate phenotype (2 µg/mL < MIC/MBC < 4 µg/mL) and another 12.5% were resistant (MIC/MBC > 4 µg/mL). Intermediate phenotypes were recovered from the jejunum and ileum, whereas resistant isolates were found in the ileum and colon. These findings suggest that the porcine colon may serve as an intestinal reservoir for C. suis, while the ileum supports more robust bacterial replication. Overall, these data contribute to our understanding of the intestinal ecology of C. suis under field conditions and its tetracycline susceptible patterns. Full article

Human iPSC-derived cardiomyocytes (iPSC-CMs) exhibit fetal-like mitochondrial networks and limited oxidative metabolism, constraining their translational utility. The key bottleneck is mitochondrial immaturity, resulting from blunted PGC-1α–NRF1/2–TFAM axis activation and insufficient nuclear–mitochondrial coordination, rather than sarcomeric or electrophysiological immaturity alone. This review synthesizes genome-guided interventions (CRISPRa and mtDNA editing) and complementary environmental strategies—including metabolic substrate switching, electromechanical stimulation, and extracellular vesicle (EV)-mediated mitochondrial transfer—to drive mitochondrial biogenesis and maturation in iPSC-CMs. We systematically reviewed studies (2005–2025) targeting (1) key regulators of mitochondrial biogenesis (PGC-1α, NRF1/2, TFAM), (2) CRISPR-based transcriptional activators/repressors and mtDNA editors (DdCBE, mitoTALENs), and (3) maturation approaches such as metabolic conditioning, electromechanical stimulation, 3D tissue culture, and EV-mediated mitochondrial transfer. CRISPRa-mediated activation of PGC-1α, NRF1, and GATA4, combined with mtDNA base editors, enhances mitochondrial mass and OXPHOS function, while integration with environmental maturation strategies further promotes adult-like phenotypes. Integrative approaches that combine genome-guided interventions (CRISPRa, mtDNA editing) with environmental maturation cues yield the most adult-like iPSC-CM phenotypes reported to date. CRISPR-guided mitochondrial biogenesis thus represents a frontier for producing metabolically competent, structurally mature iPSC-CMs for disease modeling and therapy. Remaining translational challenges include efficient mitochondrial delivery, metabolic homeostasis, and multi-omics validation. We propose standardized workflows to couple nuclear and mitochondrial editing with maturation strategies. Full article

Esteya vermicola is a nematophagous fungus with strong parasitic ability against the pinewood nematode (Bursaphelenchus xylophilus) and shows great potential for the biological control of pine wilt disease. However, this fungus is highly sensitive to environmental stress factors and often exhibits early necrosis when cultured on conventional nutrient-rich media, limiting its large-scale application. In this study, we optimized the long-term cultivation and conidiation conditions of E. vermicola CBS115803 by supplementing minimal medium (MM) with amino acids, and evaluated its stress tolerance and infectivity against the pinewood nematode. Among 20 tested amino acids, histidine significantly increased total conidia production, while arginine, glutamine, and proline markedly promoted the formation of lunate conidia. The combination of arginine, histidine, glutamine, and proline (AHGP) produced the highest overall conidia yield and lunate conidia proportion. The MM + AHGP medium maintained long-term colony viability, whereas colonies on PDA and CM media showed obvious degeneration. This formulation also improved mycelial growth, total conidiation, and the proportion of lunate conidia. Moreover, conidia produced on MM + AHGP exhibited the highest germination rates and infectivity under various stress conditions, including cold, heat, oxidative, osmotic, and UV stresses. Conidia germination was significantly enhanced following treatment at 0 °C, suggesting that low temperatures may activate dormancy-breaking pathways. This amino acid-optimized medium offers an effective technical foundation for stable large-scale production and storage of E. vermicola conidia, providing a new avenue for the biocontrol of pine wilt disease. Full article

This study examined the effects of different LED light spectra on the in vitro development of plantlets of Bletilla striata (Orchidaceae), a frost-hardy ornamental orchid with increasing horticultural relevance outside its native East Asian range. The objective was to optimize growth conditions using energy-efficient lighting to support sustainable cultivation practices. Plantlets approximately 4 cm in length with 0.5 cm leaves were cultured on standard Orchimax medium in 200 mL Erlenmeyer flasks and exposed to five LED treatments: 100% blue (B), 100% red (R), red/blue at 70:30 (RB), 50% yellow + RB (7:3), and 50% green + RB (7:3). Fluorescent light served as controls. The photosynthetic photon flux density (PPFD) was maintained at approximately 40 µmol m⁻² s⁻¹ across all light treatments. After seven weeks, selected LED spectra improved plantlet performance compared with the control. Leaf number remained stable, while RB light promoted leaf expansion, resulting in the widest leaves. Root formation occurred under all LED treatments, supporting subsequent acclimatization. Light quality strongly affected photosynthetic pigments and secondary metabolism. The highest total chlorophyll content was recorded under RB illumination (581 µg g⁻¹ FW), whereas monochromatic red light resulted in the lowest pigment levels. Carotenoid accumulation was significantly enhanced under RB and RBG spectra. Blue-containing treatments (B and RB) markedly stimulated the accumulation of phenolic compounds, including flavonols and anthocyanins, while red light suppressed phenolic biosynthesis. Total soluble sugars showed an organ-specific response, with red light promoting sugar accumulation in shoots and blue light in roots. These findings demonstrate that targeted LED lighting not only improves plant quality but also offers an environmentally sustainable and economically viable approach for commercial micropropagation and conservation of B. striata. Full article

Efficient inorganic carbon supply is a common limitation in microalgal cultivation, particularly in waste-derived media such as anaerobic digestate. Carbonic anhydrase (CA) accelerates the interconversion of CO₂ and bicarbonate and may therefore enhance carbon utilisation under conditions where inorganic carbon is abundant but not readily available. In this study, crude CA-containing extracts (aCA) were prepared from Scenedesmus-dominated algal biomass, and CA activity was quantified using an esterase assay (EAA). Although EAA activities varied depending on biomass pretreatment (0.15–0.47 U g⁻¹ DW), the physiological response to extract addition was consistent. In batch cultures of Chlorella sorokiniana grown in diluted digestate, aCA supplementation increased the specific growth rate (SGR) by 21–82%. In contrast, stimulation in a mineral medium was minimal, indicating that the benefit of aCA addition is most apparent under reduced inorganic carbon availability. In semi-continuous cultivation, repeated extract addition sustained a higher biomass productivity over time (rather than a specific growth rate). These results demonstrate that crude microalgal extracts containing CA can improve growth performance in digestate-based cultures and may offer a simple, low-cost approach to enhancing inorganic carbon utilisation in waste-integrated algal production systems. Full article

Traditional carriers play a major role in wastewater treatment worldwide due to their reliability, ease of production, well-established analytical methods, and strong treatment performance. Recent studies indicate that polyvinyl-alcohol-based microcarriers may surpass conventional media, as their smaller size, higher porosity, and increased specific surface area enable them to retain substantially more biomass within reactors. However, their practical application remains limited because fewer analytical methods and studies exist for these materials, largely due to their small dimensions and heat sensitivity, and their behaviour under industrial conditions—including their kinetics—has yet to be fully characterised and validated. This study aims to address these gaps by reviewing existing biomass measurement standards and highlighting their limitations when applied to microcarriers and by proposing alternative experimental approaches better suited for evaluating biomass on such sensitive yet high-capacity carriers. We present a set of experimental methods (still subject to further refinement) that demonstrate reliable performance with these materials, and to validate our approach, we quantified biomass in both in vitro systems and containerised-scale technologies, reaching up to 14 kg/m³ during winter and 8.7 kg/m³ in spring. Laboratory-scale experiments showed that both heterotrophic and autotrophic cultures can achieve high biomass levels of up to 21 kg/m³ and 16 kg/m³, respectively. Heterotrophs exhibited lower growth inhibition under shear stress, while autotrophs displayed a distinct shear-force niche around 0.09 µN within the reactor. Full article

Background: Serum-free culture of red blood cells (RBCs) typically leads to rapid loss of viability, limiting experimental and translational applications. Lipid-rich formulations and essential oils may provide biocompatible support for RBC integrity while limiting microbial overgrowth. Methods: RBCs from nine healthy adult donors were cultured in serum-free RPMI under four conditions: control, vehicle (olive oil, 1:100 v/v), genuine adenosine triphosphate (ATP)-oil^® (1:100 v/v), and laboratory oil, “mimicking” ATP-oil^®. Cultures were maintained for 18 days. Viability was assessed by light microscopy and trypan blue exclusion; bacterial contamination was qualitatively observed on day 18. Results: Genuine ATP-oil^® maintained 35–45% RBC viability at day 18, whereas control and vehicle cultures declined rapidly. The mimicking preparation did not reproduce these effects. ATP-oil^® immersion was associated with a qualitative reduction in bacterial contamination versus control, consistent with a dual action on RBC preservation and microbial suppression under serum-free conditions. Conclusions: Supplementation with ATP-oil^® substantially prolongs RBC survival and limits bacterial overgrowth in vitro, outperforming commonly used serum or plasma supplements on a per-volume basis. These findings suggest potential applications for improving ex vivo handling or storage of blood components and for reducing background contamination in diagnostic microbiology. Further studies with larger cohorts are warranted to reveal underlying mechanisms and to define active constituents in order to standardize production. Full article

Background/Objectives: Methicillin-resistant Staphylococcus aureus (MRSA) continues to pose a significant challenge for infection prevention, particularly because of its ability to persist on surfaces and form resilient biofilms. Although bacteriophages have attracted renewed interest as alternatives or complements to chemical disinfectants, their applied use requires careful assessment of antimicrobial performance, formulation tolerance, and genomic context. Methods: Staphylococcus-infecting bacteriophages were isolated from environmental sources and examined against reference Staphylococcus isolates. Two phage isolates, designated MRSA-W3 and SA-W2, displayed lytic activity against a broad subset of clinical MRSA strains. Using a time-resolved agar-based infection assay, phage exposure resulted in a multiplicity-of-infection-dependent decline in viable MRSA populations. Results: Time-resolved infection assays revealed a multiplicity-of-infection-dependent reduction in viable MRSA, with a pronounced decrease observed approximately 40 min post-infection. At this time point, phage-treated cultures showed a reduction of 1.2–1.8 log₁₀ CFU/mL relative to untreated controls (mean Δlog₁₀ = 1.5; 95% CI, 1.1–1.9), while control cultures remained stable. Quantitative biofilm assays demonstrated that both phages reduced biofilm biomass compared with untreated conditions, with inhibition values ranging from 20% to 45% across isolates (p ≤ 0.05), reflecting strain-dependent but reproducible effects. Assessment of formulation compatibility indicated that both phages retained infectivity following exposure to sodium dodecyl sulfate, Triton X-100, and Tween 80, whereas ethanol (≥10%) and higher concentrations of dimethyl sulfoxide were associated with rapid loss of activity. In surface disinfection models, selected phage–surfactant formulations achieved a maximum reduction of 2.18 log₁₀ CFU/cm² compared with untreated controls (p ≤ 0.05). Infection-coupled whole-genome sequencing of MRSA-infecting phage MRSA-W3 produced a high-quality assembly (99.99% completeness; 0.13% contamination) and revealed a mosaic genome containing incomplete prophage-like regions, which were interpreted conservatively as evidence of shared phage ancestry rather than active temperate behavior. Conclusions: Therefore, these findings suggest that bacteriophage-based approaches may be feasible for MRSA surface decontamination, while clearly emphasizing the need for context-specific validation before practical implementation. Full article

Background and Objectives: The COVID-19 pandemic has led to an unprecedented mental health crisis among workers in the healthcare field, with average burnout rates increasing from about 32% before the pandemic to 46–52% during peak times and post-traumatic stress disorder (PTSD) affecting 24–34% of frontline staff. The primary objective of this article is to synthesize evidence on the prevalence of burnout and PTSD among healthcare workers before and during the COVID-19 pandemic. The secondary objectives are: (a) to examine the mechanisms and empirical evidence linking clinician mental health to medical errors and patient safety outcomes and (b) to analyze the medico-legal implications of this relationship, including malpractice liability, institutional responsibility, and opportunities for policy reform. Materials and Methods: We conducted a narrative review searching PubMed (November 2025–January 2026) using predefined keyword combinations. Inclusion criteria comprised original research, systematic reviews, and meta-analyses examining mental health outcomes or patient safety among clinical staff. Data were synthesized narratively across five thematic domains. Results: Burnout prevalence increased from approximately 32% pre-pandemic to 46–52% during peak periods, with emotional exhaustion reaching 67.5% in some settings. PTSD rates rose to 24–34% among frontline staff, exceeding pre-pandemic levels of 15–20%, with ICU staff particularly affected (27–40%). Substantial overlap exists between conditions (86–98% comorbidity). Physician burnout is associated with 2.72 times higher odds of self-reported errors (95% CI: 2.19–3.37), with each point increase in emotional exhaustion raising the error risk by 5–11%. Mechanisms include cognitive impairment (reduced executive function, g = −0.39; impaired working memory, g = −0.36) and sleep disturbance. Malpractice litigation compounds psychological harm, increasing depression and suicidal ideation. Conclusions: This review, synthesizing data from over 500,000 healthcare workers, demonstrates bidirectional relationships among burnout, PTSD, and medical errors with significant medico-legal ramifications. Addressing this crisis requires systemic interventions including workload management, psychological support, blame-free reporting cultures, and policy reforms balancing accountability with recognition of system-level contributors to error. Full article

The preservation of cultural heritage within museum environments requires systematic control and monitoring of indoor microclimatic conditions. Over the past four decades, scientific evidence has established the critical role of environmental parameters, including air temperature, relative humidity, light, and airborne pollutants, in the preventive conservation of artifacts. International standards and national guidelines mandate continuous, non-invasive monitoring protocols that integrate conservation requirements with the architectural and operational constraints of historic buildings. Effective implementation necessitates a multidisciplinary approach balancing artifact preservation, human comfort, and building energy efficiency. Recent international recommendations further promote adaptive approaches wherein microclimate thresholds are calibrated to site-specific “historical climate” conditions, derived from minimum one-year baseline datasets. While essential for long-term conservation management, the design and implementation of such monitoring systems present significant technical and logistical challenges. This study presents a replicable methodological approach wherein preliminary surveys and three short-term monitoring campaigns (duration: 2 to 5 weeks) supported design, sensor selection, and spatial deployment and will allow the validation of a long-term continuous monitoring infrastructure (at least one year). These preliminary investigations enabled the following: (1) identification of priority environmental parameters; (2) optimization of sensor placement relative to exhibition layouts and maintenance protocols; and (3) preliminary assessment of microclimate risks in naturally ventilated spaces in the absence of HVAC systems. Full article

(−)-Epicatechin (EC), taxifolin (Tax), and quercetin (Q) are flavonoids with different C-ring structures. We compared their physicochemical properties and biological activities. The comparison of their stability and redox properties was conducted at mildly acidic or neutral conditions mimicking the plant vacuole or gut, and their sympathetic hyperactivation ability was examined using catecholamine (CA) excretion and blood flow. At pH 5, flavonoids were stable, but at pH 7, their retention rates decreased in the order EC > Q > Tax. LC-MS analysis detected brown oxidized oligomers in EC, while Tax and Q primarily yielded degradation products. All flavonoids exhibited O₂^•− scavenging activity after incubation at pH 5. At pH 7 after 45-min, EC and Tax promoted O₂^•−, while Q only scavenged O₂^•− after 24-hr incubation, EC’s properties decreased but Tax’s properties enhanced. Computational chemistry analysis indicated EC has higher reactivity compared to Tax and Q. EC caused a significant increase in CA excretion and blood flow, which were not observed with Tax or Q or 24-h cultured EC. These results suggest that the C-ring structure of flavonoids plays a crucial role in their stability and redox properties. Furthermore, reactive oxygen species generated by flavonoids in the gut may exert beneficial effects through sympathetic activation. Full article

Vinegar is a traditional fermented food of increasing industrial interest due to its nutritional, sensory, and bioactive properties. This study aimed to develop and optimize a controlled biotechnological process for vinegar production from the Algerian date cultivar Degla Beida, an abundant yet underexploited local resource. Indigenous Saccharomyces cerevisiae strains isolated from date fruits and Acetobacter sp. strains isolated from traditional date vinegar were employed as starter cultures in a two-stage submerged fermentation process, comprising alcoholic fermentation followed by acetic fermentation. Process optimization was carried out using Response Surface Methodology (RSM) based on a Central Composite Design (CCD), evaluating the effects of initial alcoholic degree (4–10% v/v) and yeast extract supplementation (0.2–0.5 g/L). The statistical models showed excellent fitting and predictive reliability (p < 0.0001; R² = 94.1–99.1%). Under optimal conditions (7% v/v initial alcohol, 0.2 g/L yeast extract, 30 °C, pH 5), the process yielded a maximum acetic acid concentration of 72 g/L after 11 days, with 80% fermentation efficiency and complete ethanol depletion. The optimized vinegar exhibited enhanced bioactive properties, with a total phenolic content of 620 mg GAE/100 mL and a DPPH radical scavenging activity of 78%, significantly higher than those of the unfermented juice. These results demonstrate the suitability of Degla Beida dates for vinegar production and highlight the potential of indigenous microbial resources for the sustainable valorization of local raw materials through controlled fermentation processes. Full article

Background: Porphyromonas gingivalis outer membrane vesicles (OMVs) are key mediators of host–pathogen interactions and have been implicated in both periodontal disease and systemic conditions, including pregnancy complications. Although OMV production and cargo are known to be influenced by environmental stress, how oxidative stress reshapes P. gingivalis OMVs and their functional impact on trophoblast cells remains poorly understood. Here, we investigated how exposure to hydrogen peroxide (H₂O₂) affects OMV biogenesis, composition, and their ability to modulate bacterial invasion in trophoblast cells. Methods: P. gingivalis was cultured anaerobically and exposed to 30 mM H₂O₂ during the final 24 h of growth. OMVs were isolated by differential ultracentrifugation and characterized by nanoparticle tracking analysis and transmission electron microscopy and OMV protein cargo was analyzed by proteomics. Functional effects were assessed using invasion and persistence assays in HTR-8/SVneo trophoblast cells pretreated with OMVs. Results: Oxidative stress did not significantly alter total OMV yield but resulted in smaller vesicles (control OMV 168.2 ± 8.7 nm vs. OMV from H₂O₂-treated cultures 130.0 ± 13.8 nm) with reduced negative surface charge and increased membrane-associated FM4-64 fluorescence. Proteomic analysis revealed a remodeling of the OMV protein cargo under oxidative stress, including the selective enrichment of a von Willebrand factor type A domain-containing protein. Functionally, OMVs from control cultures led to a 2.5-fold increase in P. gingivalis invasion and a 4-fold increase in intracellular persistence in trophoblast cells, whereas OMVs produced under oxidative stress failed to promote these processes. Conclusions: Together, these findings highlight oxidative stress as a key determinant of OMV-mediated host–pathogen interactions at the maternal–fetal interface. Full article