Search Results (273)

We present a miniaturized, inexpensive, and user-friendly microfluidic platform to support biological applications. The system integrates a mini-incubator providing controlled environmental conditions and housing a microfluidic device for long-term cell culture experiments. The incubator is designed to be compatible with standard inverted optical microscopes and Raman spectrometers, allowing for the non-invasive imaging and spectroscopic analysis of cell cultures in vitro. The microfluidic device, which reproduces a dynamic environment, was optimized to sustain a passive, gravity-driven flow of medium, eliminating the need for an external pumping system and reducing mechanical stress on the cells. The platform was tested using Raman analysis and adherent tumoral cells to assess proliferation prior and subsequent to hydrogen peroxide treatment for oxidative stress induction. The results demonstrated a successful adhesion of cells onto the substrate and their proliferation. Furthermore, the platform is suitable for carrying out optical monitoring of cultures and Raman analysis. In fact, it was possible to discriminate spectra deriving from control and hydrogen peroxide-treated cells in terms of DNA backbone and cellular membrane modification effects provoked by reactive oxygen species (ROS) activity. The 800–1100 cm⁻¹ band highlights the destructive effects of ROS on the DNA backbone’s structure, as its rupture modifies its vibration; moreover, unpaired nucleotides are increased in treated sample, as shown in the 1154–1185 cm⁻¹ band. Protein synthesis deterioration, led by DNA structure damage, is highlighted in the 1257–1341 cm⁻¹, 1440–1450 cm⁻¹, and 1640–1670 cm⁻¹ bands. Furthermore, membrane damage is emphasized in changes in the 1270, 1301, and 1738 cm⁻¹ frequencies, as phospholipid synthesis is accelerated in an attempt to compensate for the membrane damage brought about by the ROS attack. This study highlights the potential use of this platform as an alternative to conventional culturing and analysis procedures, considering that cell culturing, optical imaging, and Raman spectroscopy can be performed simultaneously on living cells with minimal cellular stress and without the need for labeling or fixation. Full article

The selection of start-ups by business incubators (BIs) is one of the main processes of these organizations that aim to promote entrepreneurship and economic development. Through a systematic literature review of the BI selection phase, following the PRISMA guidelines, a new conceptual model is proposed, delineating findings across three dimensions: the incubatee–incubator alignment and BI strategy, the selection criteria considered, and the decision-making methods used. The conceptual model proposed here represents the first conceptual model focused exclusively on the selection of start-ups by BIs, mapping practices in terms of start-up selection that hold important practical implications for BI managers. Considering the growing need to align economic performance with environmental and social objectives, the start-up selection process by BIs also plays a key role in fostering sustainability-oriented ventures. This fact increases the relevance of this selection phase, not only in terms of operational efficiency, but also as a potential lever for sustainable regional development. Findings emphasize the need for future research that integrates all three dimensions simultaneously, explains the selection process at an operational level, and assesses the importance of this phase for BI performance. Full article

To promote environmentally sustainable remediation and resource recovery from ion-adsorption rare earth tailings (IRET), this study comprehensively investigated the previously isolated strain REO-01 by examining its sulfate-reducing performance, Cd(II) immobilization potential, and physiological and biochemical responses under varying environmental conditions. Strain REO-01 was identified as a Gram-negative facultative anaerobe with strong sulfate-reducing activity and effective Cd(II) immobilization capacity. During a 96 h incubation period, the strain entered the exponential growth phase within 36 h, after which the OD₆₀₀ values plateaued. Concurrently, the culture pH increased from 6.83 to 7.5, and the oxidation-reduction potential (ORP) declined to approximately −300 mV. Cd(II) concentrations decreased from 0.2 mM to 3.33 μM, corresponding to a removal efficiency exceeding 95%, while sulfate concentrations declined from 1500 mg/L to 640 mg/L, with a maximum reduction efficiency of 66.16%. The strain showed optimal growth at 25–40 °C and near-neutral pH (6–7), whereas elevated Cd(II) concentrations (≥0.2 mM) significantly inhibited cell growth. A sulfate concentration of 1500 mg/L was found to be optimal for cellular activity. Among the tested carbon sources, sodium lactate at 4.67 g/L yielded the most favorable results, reducing ORP to −325 mV, increasing pH to 7.6, and lowering Cd(II) and sulfate concentrations to 3.33 μM and 510 mg/L, respectively. These findings highlight the strong potential of strain REO-01 for simultaneous sulfate reduction and Cd(II) remediation, supporting its application in the in situ bioremediation and resource utilization of rare earth tailings. Full article

The addition of alkaline amendments is considered an important strategy to alleviate soil acidification, with profound impacts on soil nitrogen (N) transformations such as nitrification as well as greenhouse gas (GHG) nitrous oxide (N₂O) emissions. Nitrification inhibitors (NIs) have been widely recognized to effectively mitigate N₂O emissions by depressing the nitrification process. However, the effectiveness of NIs on N₂O emissions reduction under different alkaline amendments remains largely unknown, hindering our knowledge of the optimal soil acidification mitigation strategies. In this study, the effects of NIs in combination with different alkaline amendments on N₂O emissions were assessed on typical acid soils collected from four sites during a 28-day aerobic incubation experiment. Treatments included four alkaline amendments (quicklime, chicken manure, cow dung, biochar) and no amendment control, designated as CaO, CM, CD, BC, and CK, combined with a typical NI (3,4 dimethylpyrazole phosphate, DMPP) applied at 2 mg soil kg⁻¹ or non-NI applied, respectively. Both individual amendments and their combination with DMPP significantly elevated the soil pH by 4.9–64.2% compared with the CK treatment, with the effectiveness ranking as CaO > CM ≈ CD > BC. Cumulative N₂O emissions were stimulated by the individual application of CaO, CM, and CD but were reduced by BC application compared with the CK treatment. Changes in N₂O emissions were positively correlated with the responses of the net N mineralization and nitrification rates to individual amendments, which were regulated by changes in the soil pH. The suppressive effects of NI combined with individual amendments on N₂O emissions were significant in the CaO treatment with a reduction ranging from 3.3% to 60.2%, which was attributed to decreased abundances of ammonia-oxidizing bacteria (AOB). Therefore, we concluded that the combined application of CaO and DMPP could be considered as a suitable mitigation strategy for addressing soil acidification through optimized N management. Additionally, BC can serve as a supplementary practice to simultaneously improve soil fertility. These insights are crucial for developing integrated fertilization management strategies to mitigate soil acidification with low N loss risks. Full article

Traditional hazard identification techniques for Vibrio parahaemolyticus often neglect the distinction between viable and nonviable bacteria in aquatic products, leading to overestimated disease risks and uncertainties in risk assessments. To address this limitation, we developed an automated PMA pretreatment instrument that integrates dark incubation and photo-crosslinking into a unified workflow, allowing customizable parameters such as incubation time, light exposure duration, and mixing speed while maintaining stable temperatures (<±1 °C fluctuation) to preserve bacterial DNA integrity. Leveraging this system, a duplex qPCR assay was optimized for simultaneous quantitative detection of V. parahaemolyticus and V. cholerae in aquatic products and environmental samples. The assay demonstrated robust performance with 90–110% amplification efficiencies across diverse matrices, achieving low limits of detection (LODs) of 10¹–10² CFU/mL in shrimp farming environment water and 10²–10³ CFU/g in shrimp (Litopenaeus vannamei) and oyster (Crassostrea gigas). Notably, it effectively discriminated viable bacteria from 10⁶ CFU/mL(g) nonviable cells and showed strong correlation with ISO-standard methods in real-world sample validation. This integrated platform offers a rapid, automated solution for accurate viable bacterial quantification, with significant implications for food safety, pathogen surveillance, and risk management in aquatic industries. Full article

African swine fever virus (ASFV), a highly contagious and lethal virus affecting domestic and wild pigs, has raised global concerns due to its continued spread across Europe and Asia. While traditional transmission pathways involve suids and soft ticks, this study investigates the potential role of freshwater gastropods as environmental reservoirs capable of sustaining ASFV. We analysed ASFV survival in ten gastropod species after long-term co-incubation with the virus. Viral transcriptional activity, particularly of the late gene B646L and members of the multigene family MGF505, was evaluated in snail faeces up to nine weeks post-infection. Results revealed that several gastropods, including Melanoides tuberculata, Tarebia granifera, Physa fontinalis, and Pomacea bridgesii, support long-term persistence of ASFV, accompanied by increased MGF505 gene expression. Notably, the simultaneous activation of MGF5052R and MGF50511R significantly correlated with higher B646L expression and extended viral survival, suggesting a functional role in ASFV maintenance. Conversely, antiviral (AV) activity assays showed that some gastropod faeces reduced replication of the unrelated Influenza virus, hinting at induced host defences. A negative correlation was observed between AV activity and the expression of MGF505 2R/11R, implying that ASFV may suppress antiviral responses to facilitate persistence. These findings suggest that certain gastropods may serve as overlooked environmental hosts, contributing to ASFV epidemiology via long term viral shedding. Further research is needed to clarify the mechanisms underlying ASFV–host interactions and to assess the ecological and epidemiological implications of gastropods in ASFV transmission cycles. Full article

An attempt was made to simulate the conditions prevailing in an agricultural crop to investigate whether and how geotextile microplastics alter the movement and accumulation of heavy metals in plants. For this purpose, a pot experiment, lasting 149 days, was carried out on soil obtained from a rural area, where pieces of a geotextile in mesoplastic dimensions, of the same chemical composition as that used by farmers in the Greek countryside, were added. Furthermore, metal solutions (Cu, Zn, Cd) were incorporated in the pots at two levels, and incubation prior to planting was carried out for two weeks. Then, industrial hemp was cultivated, while continuous measurements of its horticultural characteristics and of the levels of metals moved from the soil to the plant were made. The plants appeared to be highly resistant to the rather harsh growing conditions, and furthermore, it was observed that the cumulative metal capacity of cannabis was enhanced in most cases. The simultaneous presence of metals and geotextile (plastic) fragments enhanced the amount of Zn and Cd transfer into the soil-to-plant system. Hemp plants exhibited strong resilience abilities in the particularly stressful soil environment, possibly developing defense mechanisms. The experiments are particularly encouraging as they prove that simple and habitual practices in cultivated soils that lead to post-weather erosion of the geotextile may contribute positively in terms of remediation methods for heavy-metal-laden soils, as they indirectly help the plant to remove larger amounts of metal elements. The experiments should be intensified on a wider range of soils of different soil reactions and particle sizes and, of course, should be carried out under real field conditions in Mediterranean soil environments. Full article

Orthoherpesviridae is a large family of enveloped DNA virus. Among the most significant animal-infecting viruses are bovine alphaherpesvirus 1 (BoAHV1), caprine alphaherpesvirus 1 (CpAHV1) and equid alphaherpesvirus 1 (EqAHV1). Research into new methods to combat herpesvirus infections is ongoing. The aim of this study was to evaluate the antiviral activity of three extracts of the Helleborus bocconei roots against BoAHV1, CpAHV1 and EqAHV1. The roots were air-dried, extracted with methanol (MeOH) and then partitioned between n-butanol (n-BuOH) and water. All three extracts were tested for cytotoxicity on MDBK and RK-13 cells, and for antiviral activity. Two non-cytotoxic concentrations were assessed for their anti-BoAHV1, anti-CpAHV1 and anti-EqAHV1effects. Cells were incubated with the extracts for 72 h under three experimental conditions: pretreatment before viral infection, treatment post virus infection and simultaneous viral infection and treatment with extracts. The n-BuOH extract (BE) at 0.62 µg/mL inhibited the cytopathic effects of all three viruses in the simultaneous assay. Additionally, no cytopathic effect was observed in MDBK cells infected with CpAHV1and treated with 0.31 µg/mL BE post virus infection. Therefore, the BE contains molecules or groups of molecules potentially useful for developing an alternative therapy against herpesvirus (HV) infection. Full article

Elevated levels of homocysteine in the blood plasma (hyperhomocysteinemia, HHCY) positively correlate with migraine symptoms in patients. Experimental studies show a higher sensitivity of rats with prenatal HHCY (pHHCY) to migraine symptoms like allodynia, photophobia, anxiety, and a higher excitability of meningeal trigeminal afferents. In the present study, the roles of purinergic mechanisms in the homocysteine-induced hyperexcitability of the trigeminal ganglion (TG) system using electrophysiological recordings from the trigeminal nerve, Ca²⁺ imaging of cells isolated from TG, and mast cell staining in meninges were investigated. Experiments were performed using rats with pHHCY born from females fed with a high-methionine-containing diet before and during pregnancy. Firstly, we found that lower concentrations of 4-aminopyridine, a K⁺-channel blocker, were able to induce an increase in the nociceptive activity of trigeminal afferents, supporting the hypothesis of the higher excitability of the trigeminal nerve of rats with pHHCY. Trigeminal afferents of rats with pHHCY were more sensitive to the exogenous application of the nonspecific agonist of purinergic ATP receptors. In neurons and satellite glial cells of TG of rats with pHHCY ATP, ADP (an agonist of metabotropic P2Y receptors) and BzATP (an agonist of ionotropic P2X with especially high potency for the P2X7 receptor) induced larger Ca²⁺ transients. The incubation of TG neurons in homocysteine for 24 h increased the ratio of neurons responding simultaneously to ATP and capsaicin. Moreover, rats with pHHCY exhibit a higher rate of degranulation of mast cells and increased response to the agonist of the P2X7 receptor BzATP application. In addition, higher levels of calcitonin gene-related peptide (CGRP) were found in rats with pHHCY. Our results suggest that chronic elevated levels of homocysteine induce the upregulation of ionotropic or metabotropic ATP receptors in neurons, satellite glial cells, and mast cells, which further provide inflammatory conditions and the sensitization of peripheral afferents underlying pain. Full article

The objective was to determine the digestible indispensable amino acid score (DIAAS) of food sources using the pig model and in vitro procedures. The standardized ileal digestibility (SID) of amino acids (AA) in white rice, wheat, pea protein concentrate (PPC), soy protein isolate (SPI), and skim milk powder (SMP) were determined in pigs. Twelve barrows with an initial body weight of 28.4 kg (standard deviation = 4.0) equipped with a T-cannula at the distal ileum were allotted to a replicated 6 × 5 incomplete Latin square design with six diets and five periods. Five experimental diets contained each ingredient as the only source of AA. Additionally, a nitrogen-free diet was prepared to measure the basal endogenous losses of AA for the calculation of the SID of AA. The in vitro ileal disappearance (IVID) of crude protein (CP) in the five food sources was also determined using the conventional flask procedure and the multi-sample simultaneous in vitro procedure employing the Daisy^II incubator. The DIAAS values for white rice were 42, 50, and 60 for infants, children, and adults, respectively. For wheat, the DIAAS values were 46, 56, and 66, respectively. The DIAAS values for PPC were 47, 58, and 69 for infants, children, and adults, respectively. For SPI, the values were 61, 74, and 87, respectively. The DIAAS for SMP were 82, 112, and 131 for infants, children, and adults respectively. The IVID of CP in SMP was greater (p < 0.05) than that in other sources. The DIAAS calculated based on IVID of CP were similar to those determined based on the SID of AA in pigs. In conclusion, white rice, wheat, pea protein concentrate, and soy protein isolate were observed to have DIAAS values less than 100, indicating that these ingredients need to be combined with other protein sources to meet the indispensable AA requirements. Skim milk powder had a DIAAS greater than 100 for children and adults. Full article

This study presents the development of an innovative electrochemical sensor for the simultaneous detection of cadmium (Cd²⁺) and lead (Pb²⁺) ions in environmental samples. The sensor is developed based on a composite material of zeolite imidazolate framework ZIF-7 and polyaniline (PANI), referred to as ZIF-7@PANI, where ZIF-7 is rapidly synthesized at room temperature and polyaniline used to enhance the conductivity of the composite. Characterization via X-ray diffraction (DRX), scanning electron microscopy (SEM), and Fourier transform infrared spectroscopy (FTIR) confirmed successful synthesis. The composite was applied to a glassy carbon electrode (GCE) using drop-casting for heavy metal ion detection. Experimental parameters—including pH, incubation time, deposited quantity, and drying time—were optimized using the Box–Behnken design. Under optimal conditions, the ZIF-7@PANI/GCE sensor demonstrated a broad dynamic concentration range (0.002–1 µM for Pb²⁺ and 0.02–30 µM for Cd²⁺), with low detection limits (2.96 nM for Pb²⁺ and 10.6 nM for Cd²⁺). It also exhibited strong anti-interference properties and high recovery rates (85–110%), highlighting its potential for real practical applications. Full article

Garlic (Allium sativum L.), a vital global crop, suffers significant losses from soil-borne fungal pathogens such as Fusarium proliferatum, responsible for Fusarium bulb rot, and Stromatinia cepivora, the causal agent of white rot. In May 2023, garlic fields near Makó City, Hungary, showed simultaneous yellowing and wilting symptoms of unknown fungal infestations, which appeared sporadically. The causal pathogens were later confirmed as F. proliferatum and S. cepivora through sampling of symptomatic garlic plants, incubation in humid chambers to stimulate fungal growth, and culturing on Potato Dextrose Agar (PDA) under sterile conditions. This was followed by hyphal tip isolation and purification. Molecular identification was performed using ITS1-2 sequencing, supported with morphological identification based on colony and microscopic features. This research aimed to elucidate pathogen interaction dynamics and assess the resistance of eleven garlic cultivars to both single and simultaneous inoculations by these pathogens, under in vitro and in planta tests. Dual culture assays of F. proliferatum and S. cepivora were studied at two time points: Day 8, marking the cessation of growth along the interacting fronts due to competitive coexistence, and Day 14, when single cultures reached full radial growth. On Day 8, inhibition percentages were 8.47% for F. proliferatum and 6.40% for S. cepivora, reflecting the initial effects of competitive interactions at the point of contact. By Day 14, inhibition rates increased to 25.39% and 28.61%, respectively, highlighting the cumulative effects of sustained competition and the growing difference between single and dual culture growth. Inoculation trials, involving placing fungal disks onto the basal areas of wounded garlic cloves, revealed considerable variability in disease incidence and severity. Cultivars such as ‘Aulxito’, ‘Sabadrome’, ‘Arno’, ‘Garcua’, and ‘Makói Tavaszi’ were highly susceptible to both pathogens, while ‘Flavor’ and ‘Sabagold’ exhibited only mild symptoms when inoculated with F. proliferatum and S. cepivora, respectively. Simultaneous inoculation resulted in more rapid and severe infections, exhibiting disease incidences above 96.00% across all cultivars. Remarkably, the cultivar ‘Elephant’ exhibited complete resistance to both pathogens, even under simultaneous inoculation, highlighting its potential for future garlic resistance breeding programs. Full article

Objectives: To develop and evaluate graphene oxide/gelatin/alginate scaffolds for advanced wound therapy capable of mimicking the native extracellular matrix (ECM) and bio-stimulating all specific phases of the wound healing process, from inflammation and proliferation to the remodeling of damaged skin tissue in three dimensions. Methods: The scaffolds were engineered as interpenetrating polymeric networks by the crosslinking reaction of gelatin in the presence of alginate and characterized by structural, morphological, mechanical, swelling properties, porosity, adhesion to the skin tissue, wettability, and in vitro simultaneous release of the active agents. Biocompatibility of the scaffolds were evaluated in vitro by MTT test on fibroblasts (MRC5 cells) and in vivo using Caenorhabditis elegans assay. Results: The scaffolds exhibited a highly porous interconnected morphology with adjustable porosity (93–96%) and mechanical strength (1.10–2.90 MPa), hydrophilic nature with high capacity to absorb physiological fluids, and stable adhesion to the skin tissue. The obtained results of MRC5 cell viability indicate that the scaffolds are safe for biomedical applications. No mortality was detected among the Caenorhabditis elegans throughout the incubation period, indicating that the scaffolds are not toxic. The results of in vitro release study of allantoin, quercetin, and caffeic acid confirm the scaffolds’ significant potential for simultaneous release. Conclusion: The graphene oxide/gelatin/alginate scaffolds are promising candidates for non-invasive, dual ECM-mimetic, and multi-target wound therapy, offering an innovative strategy to address the complexities of wound healing process. Full article

Selol is a semi-synthetic mixture of selenized triglycerides. The results of biological studies revealed that Selol exhibits several anticancer effects. However, studies on its potential anti-inflammatory activity are scarce, and underlying signaling pathways are unknown. The aim of our study was to investigate the ability of Selol to exert anti-inflammatory effects in a RAW 264.7 cell line model of LPS (lipopolysaccharide)-induced inflammation. Cells were treated either with Selol 5% (4 or 8 µg Se/mL) or LPS (1 µg/mL) alone or with Selol given concomitantly with LPS. The parameters studied were reactive oxygen species (ROS) production, glutathione and thioredoxin (Txn) levels, and nuclear factor kappa B (NF-κB) activation, as well as nitric oxide/prostaglandin E₂ (NO/PGE₂) production. The presented research also included the effect of Selol and/or LPS on glucose (Glc) catabolism; for this purpose, the levels of key enzymes of the glycolysis pathway were determined. The results showed that Selol exhibited pro-oxidative properties. It induced ROS generation with a significant increase in the level of Txn; however, it did not affect the reduced glutathione/oxidized glutathione (GSH/GSSG) ratio. Selol moderately activated NF-κB but failed to affect NO/PGE₂ production. The effect of Selol on glucose catabolism was not significant. However, the simultaneous administration of Selol with LPS exerted a statistically significant anti-inflammatory effect via a decrease in the production of pro-inflammatory mediators and NF-κB activation. Our study also showed that as a result of LPS action in cells, the anaerobic glycolysis activity was increased, and incubation with Selol caused a partial reprogramming of Glc metabolism towards aerobic metabolism. This may indicate different pharmacological and molecular effects of Selol action in physiological and pathological conditions. Full article

The resistance of 16 Rhodococcus strains to diesel fuel was studied. The minimal inhibitory concentrations of diesel fuel against Rhodococcus were 4.0–64.0 vol. % and 0.5–16.0 vol. % after 7 days of incubation in Luria–Bertani broth and a mineral “Rhodococcus-surfactant” medium, respectively. The three most resistant strains (R. ruber IEGM 231, IEGM 442 and Rhodococcus sp. IEGM 1276) capable of overcoming the toxicity of diesel fuel at a high (8.0 vol. %) concentration and at a low (4 °C) temperature were selected. Respiration activities, growth kinetics, and changes in the diesel fuel composition during the biodegradation process were elucidated using gas chromatography with mass spectrometry, respirometry, and Bradford analysis. Growth conditions were optimised for the improved biodegradation of diesel fuel by Rhodococcus cells using multifactor analysis. They included the simultaneous addition of 1.3 g·L⁻¹ of granular sugar and 0.25 g·L⁻¹ of yeast extract. The twofold stimulation of the biodegradation of individual hydrocarbons in diesel fuel (n-pentadecane, n-hexadecane and n-heptadecane) was demonstrated when glycolipid Rhodococcus-biosurfactants were added at a concentration of 1.4 g·L⁻¹. A total removal of 71–91% of diesel fuel was achieved in this work. Full article