Search Results (883)

Ex situ conservation is a vital strategy of preserving plant species at risk, offering practical methods to obtain information regarding species-specific germination characteristics. Campanula pangea, a local endemic species of NE Greece, has been previously classified as vulnerable, partly due to the lack of knowledge about its biology. This study focused on the germination behaviour of C. pangea stored seeds by assessing their germination success under the effects of incubation temperature and gibberellic acid (GA₃). To contextualize the experimental conditions, a bioclimatic profile of the species was developed using open-access temperature and precipitation data that characterize its natural habitat. The results showed that the optimal germination temperature range for C. pangea is 15–20 °C. Pre-treatment of seeds with GA₃ solution (1000 mg L⁻¹) widened the germination range of the seeds only at the low temperature of 10 °C. The experimentation results showed that the seeds of C. pangea exhibit dormancy. These findings contribute to the development of a species-specific germination protocol for ex situ propagation and conservation, enhance understanding of the species’ germination requirements, and thus support future conservation efforts and assessments of extinction risk, or other ornamental applications and/or targeted medicinal research. Full article

Background: Obesity is associated with insulin resistance (IR) and characterized by impaired activation of the PI3K/AKT route and glucose uptake. Elevated plasma levels of palmitic acid (PA) diminish insulin signaling in vitro and in vivo. Origanum vulgare L. essential oil (OVEO) is rich in monoterpenes with protective effects against IR. Objective: The study aimed to assess total phenols content and antioxidant activity of OVEO and its cytotoxicity, as well as its effect on insulin signaling and glucose uptake in PA-treated adipocytes. Methods: The quantification of total phenolic content was determined using the Folin–Ciocalteu method, while the antioxidant capacity of OVEO was assessed by DPPH (2,2-diphenyl-1-picrylhydrazyl) and FRAP (ferric reducing antioxidant power) methods. The cytotoxicity of OVEO (0.1–10 µg/mL) was assessed using the MTS assay. SW872 adipocytes were incubated with 0.4 mM PA for 24 h, with or without a 2 h preincubation of OVEO, and then stimulated with insulin (100 nM, 10 min) or a vehicle. Phosphorylation of Tyr-IRS-1, Ser-AKT, and Thr-AS160 was analyzed by Western blot, and glucose uptake was measured using 2-NBDG. Results: OVEO contained phenols and exhibits antioxidant capacity. All the concentrations of OVEO assessed were not cytotoxic on SW872 adipocytes. PA decreased basal phospho-AS160 as well as insulin-stimulated phospho-IRS1, phospho-AKT, phospho-AS160 and glucose uptake, while OVEO co-treatment enhanced these markers. Conclusions: These findings suggest a beneficial effect of OVEO on the PA-impaired insulin pathway and glucose uptake, which might be explained by its phenolic content and antioxidant capacity, highlighting its potential as a complementary therapeutic agent for IR and related metabolic disorders. Full article

A novel cellulose-degrading bacterial strain, D3-1, capable of degrading cellulose under medium- to high-temperature conditions, was isolated from soil samples and identified as Staphylococcus caprae through 16SrRNA gene sequencing. The strain’s cellulase production was optimized by controlling different factors, such as pH, temperature, incubation period, substrate concentration, nitrogen and carbon sources, and response surface methods. The results indicated that the optimal conditions for maximum cellulase activity were an incubation time of 91.7 h, a temperature of 41.8 °C, and a pH of 4.9, which resulted in a maximum cellulase activity of 16.67 U/mL, representing a 165% increase compared to pre-optimization levels. The above experiment showed that, when maize straw flour was utilized as a natural carbon source, strain D3-1 exhibited relatively high cellulase production. Furthermore, gas chromatography–mass spectrometry (GC-MS) analysis of products in the degradation liquid revealed the presence of primary sugars. The results indicated that, in the denitrification of simulated sewage, supplying maize straw flour degradation liquid (MSFDL) as the carbon source resulted in a carbon/nitrogen (C/N) ratio of 6:1 after a 24 h reaction with the denitrifying strain WH-01. The total nitrogen (TN) reduction was approximately 70 mg/L, which is equivalent to the removal efficiency observed in the glucose-fed denitrification process. Meanwhile, during a 4 h denitrification reaction in urban sewage without any denitrifying bacteria, but with MSFDL supplied as the carbon source, the TN removal efficiency reached 11 mg/L, which is approximately 70% of the efficiency of the glucose-fed denitrification process. Furthermore, experimental results revealed that strain D3-1 exhibits some capacity for nitrogen removal; when the cellulose-degrading strain D3-1 is combined with the denitrifying strain WH-01, the resulting TN removal rate surpasses that of a single denitrifying bacterium. In conclusion, as a carbon source in municipal sewage treatment, the degraded maize straw flour produced by strain D3-1 holds potential as a substitute for the glucose carbon source, and strain D3-1 has a synergistic effect with the denitrifying strain WH-01 on TN elimination. Thus, this research offers new insights and directions for advancement in environmental sewage treatment. Full article

Oxidized phospholipids (OxPLs) are increasingly recognized as biologically active lipids involved in various pathologies. Both exposure to pathogenic factors and the efficacy of protective mechanisms are critical to disease development. In this study, we characterized an immunoassay that quantified the total capacity of the plasma to degrade or mask OxPLs, thereby preventing their interaction with cells and soluble proteins. OxLDL-coated plates were first incubated with human blood plasma or a control vehicle, followed by an ELISA using a monoclonal antibody specific to oxidized phosphatidylethanolamine. Pretreatment with the diluted blood plasma markedly inhibited mAb binding. The masking assay was optimized by evaluating the buffer composition, the compatibility with various anticoagulants, potential interfering compounds, the kinetic parameters, pre-analytical stability, statistical robustness, and intra- and inter-individual variability. We propose that this masking assay provides a simple immunological approach to assessing protective mechanisms against lipid peroxidation products. Establishing this robust and reproducible method is essential for conducting clinical association studies that explore masking activity as a potential biomarker of the predisposition to a broad range of lipid-peroxidation-related diseases. Full article

Several species of Fusarium are reported to infect inflorescences of high-THC-containing cannabis (Cannabis sativa L.) plants grown in greenhouses in Canada. These include F. graminearum, F. sporotrichiodes, F. proliferatum, and, to a lesser extent, F. oxysporum and F. solani. The greatest concern surrounding the infection of cannabis by these Fusarium species, which cause symptoms of bud rot, is the potential for the accumulation of mycotoxins that may go undetected. In the present study, both naturally infected and artificially infected inflorescence tissues were tested for the presence of fungal-derived toxins using HPLC-MS/MS analysis. Naturally infected cannabis tissues were confirmed to be infected by both F. avenaceum and F. graminearum using PCR. Pure cultures of these two species and F. sporotrichiodes were inoculated onto detached inflorescences of two cannabis genotypes, and after 7 days, they were dried and assayed for mycotoxin presence. In these assays, all Fusarium species grew prolifically over the tissue surface. Tissues infected by F. graminearum contained 3-acetyl DON, DON, and zearalenone in the ranges of 0.13–0.40, 1.18–1.91, and 31.8 to 56.2 μg/g, respectively, depending on the cannabis genotype. In F. sporotrichiodes-infected samples, HT2 and T2 mycotoxins were present at 13.9 and 10.9 μg/g in one genotype and were lower in the other. In F. avenaceum-inoculated tissues, the mycotoxins enniatin A, enniatin A1, enniatin B, and enniatin B1 were produced at varying concentrations, depending on the isolate and cannabis genotype. Unexpectedly, these tissues also contained detectable levels of 3-acetyl DON, DON, and zearalenone, which was attributed to apre-existing natural infection by F. graminearum that was confirmed by RT-qPCR. Beauvericin was detected in tissues infected by F. avenaceum and F. sporotrichiodes, but not by F. graminearum. Naturally infected, dried inflorescences from which F. avenaceum was recovered contained beauvericin, enniatin A1, enniatin B, and enniatin B1 as expected. Uninoculated cannabis inflorescences were free of mycotoxins except for culmorin at 0.348 μg/g, reflecting pre-existing infection by F. graminearum. The mycotoxin levels were markedly different between the two cannabis genotypes, despite comparable mycelial colonization. Tall fescue plants growing in the vicinity of the greenhouse were shown to harbor F. avenaceum and F. graminearum, suggesting a likely external source of inoculum. Isolates of both species from tall fescue produced mycotoxins when inoculated onto cannabis inflorescences. These findings demonstrate that infection by F. graminearum and F. avenaceum, either from artificial inoculation or natural inoculum originating from tall fescue plants, can lead to mycotoxin accumulation in cannabis inflorescences. However, extensive mycelial colonization following prolonged incubation of infected tissues under high humidity conditions is required. Inoculations with Penicillium citrinum and Aspergillus ochraceus under these conditions produced no detectable mycotoxins. The mycotoxins alternariol and tentoxin were detected in several inflorescence samples, likely as a result of natural infection by Alternaria spp. Fusarium avenaceum is reported to infect cannabis inflorescences for the first time and produces mycotoxins in diseased tissues. Full article

In order to recycle plastic waste back to food contact materials (FCMs), it is necessary to identify hazardous substances in plastic packaging that pose a toxicological risk. Printing inks on plastics are not yet designed to withstand the high heat stress of mechanical recycling processes and therefore require hazard identification. In this study, virgin polypropylene (PP) foils were printed with different types of inks (UV-cured, water-based) and colour shades. Thermal analysis of printed foils and pigments was performed using differential scanning calorimetry (DSC). Samples were then thermally treated below and above measured thermal events at 120 °C, 160 °C, 200 °C or 240 °C for 30 min. Subsequently, migration tests and miniaturised Ames tests were performed. Four out of thirteen printed foils and all three pigments showed positive results for mutagenicity in miniaturised Ames tests after thermal treatment at 240 °C. Additionally, pre-incubation Plate Ames tests (according to OECD 471) were performed on three pigments and one printed foil, yielding two positive results after thermal treatment at 240 °C. These results indicate that certain ink components form hazardous decomposition products when heated up to a temperature of 240 °C. However, further research is needed to gain a better understanding of the chemical processes that occur during high thermal treatment. Full article

The asynchrony of cytoplasmic and nuclear maturation in cumulus–oocyte complexes (COCs) due to prematurely declining concentrations of cyclic adenosine monophosphate (cAMP) has been shown to result in reduced oocyte developmental competence. The objective of this study was to evaluate the effect of pre-IVM treatment with cAMP modulators for different durations on the developmental potential of equine oocytes used for cloned embryo production. Collected COCs were transferred to cryovials filled with transport medium at 20–22 °C. Within the cryovials, the COCs were either untreated (Control) for 18 h or treated with 50 µM forskolin and 100 µM 3-isobutyl-1-methylxanthine for the first 4 h (Pre-IVM 4 h) or the entire 18 h (Pre-IVM 18 h). Oocytes were then transferred to maturation medium and incubated for a further 22–24 h at 38.5 °C in 5% CO₂ in air. Somatic cell nuclear transfer embryos were then produced using the meiotically mature oocytes and donor cells from six different fibroblast cell lines. The rates of maturation and embryo development did not differ significantly between the groups, though blastocyst formation tended to be inferior in the Pre-IVM 4 h group compared with the Control group (p = 0.06). Of 67 blastocysts produced, 23 were transferred to recipient mares on Day 4 or 5 post-ovulation. Regarding the pregnancy outcomes, no significant differences were found between the groups, and four viable foals were born, each derived from a different donor cell line. The findings expand on those from previous evaluations of this biphasic IVM system, and indicate that the cAMP-modulating treatments exert limited effects under the pre-IVM conditions used here. Full article

The aim of this study was to develop and characterize UV-crosslinked hydrogel matrices based on polyethylene glycol diacrylate (PEGDA), gum arabic, betaine, and sodium alginate for potential bioanalytical applications. Various physicochemical analyses were performed, including pre-polymerization emulsion stability (Multiscan), FT-IR spectroscopy, swelling behavior in physiological buffers, pH monitoring, contact angle measurements, and morphological assessment via SEM and optical microscopy. The results demonstrated that both alginate content and UV exposure time significantly influence the structural and functional properties of the hydrogels. The highest swelling ratio (2.32 g/g) was observed for the formulation containing 5% sodium alginate polymerized for 5 min (5SA_5), though this sample showed mechanical fragmentation during incubation. In contrast, the most balanced performance was achieved for the 10SA_15 formulation, which maintained structural integrity and exhibited a swelling ratio of 1.92 g/g after 9 days. The contact angle analysis revealed a surface hydrophilicity range from 50° to 100°, with the lowest angle (50°) recorded for 10SA_5, indicating high surface wettability. These findings confirm the suitability of such hydrogels for biomedical applications, particularly as absorbent, stable platforms for drug delivery or wound healing. Full article

Prion diseases are classically attributed to the accumulation of protease-resistant prion protein (PrP^Sc); however, recent evidence suggests that alternative misfolded prion conformers and systemic inflammatory factors may also contribute to neurodegeneration. This study investigated whether recombinant moPrP^Res, generated by incubating wild-type mouse PrP^C with bacterial lipopolysaccharide (LPS), can induce prion-like disease in FVB/N female mice, whether LPS alone causes neurodegeneration, and how LPS modulates disease progression in mice inoculated with the Rocky Mountain Laboratory (RML) strain of prions. Wild-type female FVB/N mice were randomized into six subcutaneous treatment groups: saline, LPS, moPrP^Res, moPrP^Res + LPS, RML, and RML + LPS. Animals were monitored longitudinally for survival, body weight, and clinical signs. Brain tissues were analyzed histologically and immunohistochemically for vacuolar degeneration, PrP^Sc accumulation, reactive astrogliosis, and amyloid-β plaque deposition. Recombinant moPrP^Res induced a progressive spongiform encephalopathy characterized by widespread vacuolation and astrogliosis, yet with no detectable PrP^Sc by Western blot or immunohistochemistry. LPS alone triggered a distinct neurodegenerative phenotype, including cerebellar amyloid-β plaque accumulation and terminal-stage spongiosis, with approximately 40% mortality by the end of the study. Co-administration of moPrP^Res and LPS resulted in variable regional pathology and intermediate survival (50% at 750 days post-inoculation). Interestingly, RML + LPS co-treatment led to earlier clinical onset and mortality compared to RML alone; however, vacuolation levels were not significantly elevated and, in some brain regions, were reduced. These results demonstrate that chronic endotoxemia and non-infectious misfolded PrP conformers can independently or synergistically induce key neuropathological hallmarks of prion disease, even in the absence of classical PrP^Sc. Targeting inflammatory signaling and toxic prion intermediates may offer novel therapeutic strategies for prion and prion-like disorders. Full article

Contamination of environmental surfaces by nosocomial pathogens like Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus), and Enterococcus spp. poses significant health risks worldwide. However, gold-standard detection methods are too time-consuming and labor-intensive. This study aimed to optimize loop-mediated isothermal amplification (LAMP) as a rapid, innovative, and cost-effective approach, comparing its effectiveness with the gold-standard cultural method. Sterile surfaces (24 cm²) were contaminated in duplicate with different concentrations of P. aeruginosa, S. aureus, and Enterococcus faecalis (E. faecalis) reference stains. For each pair of contaminated surfaces, one was analyzed using the agar contact plate method (UNI EN 17141:2021), while the other was analyzed using LAMP, following three different pre-incubation times (three, six, and nine hours). The sensitivity and accuracy of LAMP for P. aeruginosa improved with longer incubation times, reaching a value of 1.00 at nine hours, while the specificity and positive predictive value (PPV) remained at 1.00 regardless of the incubation time. For S. aureus, LAMP achieved a sensitivity, specificity, accuracy, PPV, and negative predictive value (NPV) of 1.00 across all incubation times. Finally, for E. faecalis, sensitivity increased from 0.57 at three hours to 1.00 at six and nine hours, with a high specificity, accuracy, PPV, and NPV from six hours onwards. These findings showed that LAMP can be used as a rapid and reliable alternative to gold-standard methods for detecting pathogens on surfaces. The high sensitivity and specificity achieved, especially at six and nine hours of pre-incubation, suggested its use for real-time monitoring in healthcare settings. Further research in real-world environments is needed to confirm these findings. Full article

Aspects related to the contamination of hatching eggs, sanitary management during pre-incubation, and the performance of the incubation process can compromise productive efficiency in poultry farming. When these factors negatively influence poultry farming, they can destabilize the generation and distribution of financial resources throughout the production chain, as well as limit public access to poultry-derived proteins. Understanding how these aspects are interrelated is essential for making decisions that benefit poultry health and productivity. Therefore, we conducted a multivariate analysis of microbiological and incubation parameters to evaluate whether bacterial contamination of the eggshell and yolk sac negatively affects HI and to compare the effectiveness of different sanitization protocols in reducing bacterial contamination in these regions. To achieve this, we utilized the raw data from our previous research on the sanitization of hatching eggs and conducted a detailed statistical analysis to evaluate the relationships between the studied variables. The correlation analysis revealed that eggshell mesophilic bacterial contamination (EGM) was strongly associated with yolk sac mesophilic bacterial contamination (YSM) (r = 0.76) and yolk sac contamination by Enterobacteriaceae (YSE) (r = 0.73). The principal component analysis indicated a negative association between HI performance and eggshell and yolk sac contamination. Results indicated beneficial associations between the reduction of contamination in hatching eggs and increased hatchability rates when using essential oils. The bacterial load of hatching eggs contributes to reduced productivity, reaffirming the need for proper egg sanitization, especially using essential oils. Full article

Central nervous system disorders, such as Alzheimer’s disease, Parkinson’s disease, and multiple sclerosis, are associated with complex pathophysiological mechanisms involving oxidative stress, inflammation, and protein misfolding. According to the literature, betulin, a natural compound derived from the bark of birch trees, demonstrates promising neuroprotective effects. This study investigates the neuroprotective potential of betulin and its complex with cyclodextrin, referred to as Betula Forte, using an in vitro model of differentiated SH-SY5Y neuroblastoma cells. Specifically, the study explores the antioxidant and antiapoptotic properties of these compounds under oxidative stress induced by hydrogen peroxide (H₂O₂). Our results indicated that Betula Forte exhibited lower cytotoxicity compared to betulin alone. Both substances enhanced cell viability in pre-incubation and co-incubation models, with Betula Forte showing superior efficacy under severe oxidative stress. Additionally, both substances exhibited protective effects against H₂O₂-induced oxidative stress and apoptosis, as evidenced by reduced levels of reactive oxygen species (ROS) and a lower number of apoptotic cells compared with the H₂O₂-treated cells. These findings suggest that Betula Forte, due to lower cytotoxicity, may offer a more effective neuroprotective strategy than betulin alone, highlighting its potential as a therapeutic agent for neurodegenerative diseases. Full article

Temperature is a critical factor affecting the development and population dynamics of many organisms. An organism’s ability to withstand extreme temperature events, such as heat waves, will become increasingly important as the severity, duration, and frequency of these events continue to rise worldwide due to global warming. Knowledge on the effects of heat stress on both pests and their natural enemies will thus be crucial for keeping biological control and pest control programs effective in future. This research aimed to study the effect of short-term heat stress on the predatory mite Phytoseiulus persimilis, which is one of the important natural enemies utilized as a biocontrol agent against spider mites such as Tetranychus urticae. The experiments assessed the immature developmental time of P. persimilis after a four-hour incubation of eggs at high temperatures, namely 36, 38, 40, and 42 °C, as well as 85 ± 5% RH and a 16:8 h photoperiod (L:D). After adult females emerged, they were exposed to the same conditions again and the population parameters were monitored. The results demonstrated that the immature development time decreased as temperature increased, with the shortest development duration of 5.30 days seen in eggs exposed to 40 °C, while the eggs exposed to 42 °C did not hatch. Female and male adult longevity decreased significantly as the temperature increased. Fecundity, the adult pre-ovipositional period, and the total pre-ovipositional period were lowest following the 40 °C treatment. The population parameters of P. persimilis, including r and λ, reached their highest values in mites treated at 36 °C, and were significantly higher than in the control group. Addressing these challenges through targeted research and adaptive management is essential to sustaining the efficiency of P. persimilis in biocontrol programs, particularly in the context of global climate change. Full article

Andrographolide, fucoidan, or a combination of both compounds were evaluated to determine their effects on the antiviral response in the Atlantic salmon macrophage-like cell line (SHK-1) infected with infectious pancreatic necrosis virus (IPNV). We assessed the transcript expression levels of key molecules involved in the interferon (IFN)-dependent antiviral response, as well as the viral load in cells treated with these compounds. In non-infected cells, incubation with either fucoidan, andrographolide, or a mixture of both resulted in an increase in the transcript expression of IFNα1 and various interferon-stimulated genes (ISGs). In IPNV-infected cells, treatment with either fucoidan or andrographolide separately did not significantly enhance the antiviral response compared to that of infected cells that had not previously been treated with these compounds. In contrast, the combination of andrographolide and fucoidan led to a marked increase in the transcript expression of viperin and a significant reduction in viral load. Overall, combining andrographolide and fucoidan resulted in a greater reduction in IPNV viral load in infected cells than that noted when the compounds were administered individually. Our findings suggest that pre-incubation with this mixture promotes the establishment of a protective antiviral state against IPNV, likely mediated by an IFN-dependent response. Full article

Biofouling, the undesirable deposition of microorganisms on surfaces, is ubiquitous in aqueous systems. This is no different for systems running with water-miscible metalworking fluids (MWFs), which additionally contain many organic chemicals that create favorable conditions for growth and metabolism. Biofilm formation is thus inevitable, as there is no shortage of wetted surfaces in metalworking systems. MWF manufacturers tried in vain to offer resistance by using biocides and biostatic compounds as ingredients in concentrates and as tank-side additives. We report here that such elements, alone or as components of MWFs, did not prevent biofilm formation and had negligible effects on pre-established laboratory biofilms. Moreover, biofilms in metalworking systems are interwoven with residues, sediments, and metal swarfs generated during machining. Again, co-incubation of such “real” biofilms with MWFs had no significant effect on population size—but on population composition! The implications of this finding are unclear but could provide a starting point for the treatment of biofouling, as biofilm population structure might be of importance. Finally, we show that bacteria gain function in biofilms and that they were able to degrade a toxic amine in MWFs, which the same bacteria were unable to do in planktonic form. Full article