Search Results (10,678)

Wastewater treatment plants (WWTPs) are recognized hotspots for the convergence and dissemination of antimicrobial-resistant bacteria (ARB) and antimicrobial resistance genes (ARGs) into the environment. Among ARB, carbapenem-resistant Enterobacterales (CR-E) and extended-spectrum β-lactamase-producing Escherichia coli and Klebsiella pneumoniae (ESBL-Ec/Kp) are of particular concern due to their clinical relevance. We characterized 30 CR-E and 176 ESBL-Ec/Kp isolates (two of them were both ESBL-producing and carbapenem-resistant) recovered from influent, intermediate, effluent, sludge, and downstream river samples collected from two WWTPs in northern Spain. Isolates were evaluated for resistance phenotypes against 12 antimicrobials, and β-lactamase-encoding genes were assessed by PCR and sequencing. Notably, among CR-E isolates, bla_KPC-2 was the most prevalent (93%), followed by bla_OXA-48-like, detected in two isolates from non-treated and pasteurized sludge; both isolates also carried bla_CTX-M-15, a finding not previously reported specifically in sludge samples. Among ESBL-Ec/Kp, a broad diversity of ESBL genes was identified, including bla_{CTX-M group 1} (variants 1, 3, 15, 32, 55), bla_{CTX-M group 9} (variants 14, 27, 65, 97), bla_SHV-12 and bla_TEM-169. The most prevalent ESBL gene was bla_CTX-M-15 (48.3%), followed by bla_CTX-M-14, bla_CTX-M-32, and bla_SHV-12, detected in 10.8%, 8.5%, and 6.8% of isolates, respectively. CR-E and ESBL-Ec/Kp were found in all sample types and were still detectable at terminal stages, indicating persistence throughout treatment. These findings support the need to improve and optimize current wastewater treatment methods and underscore the importance of integrating culture-based and molecular methods into routine WWTP monitoring for early detection of microbiological hazards, although further research is still needed. Full article

Methane is a potent greenhouse gas that requires its emissions to be mitigated. A significant source for methane emissions is in the form of the biogas that is produced from anaerobic digestion in wastewater reclamation and landfill facilities. Biogas has a high valorization potential in the form of its bioconversion into ectoines, an active ingredient in skin care products, by halotolerant alkaliphilic methanotrophs. Cultures of Methylotuvimicrobium alcaliphilum 20Z were grown in bench scale stirred-tank reactors to determine factors to improve methane uptake and removal. Tangential flow filtration was also implemented for a bio-milking method to recover ectoine from culture media. Methane uptake and reactor productivity increased, with a temperature of 28 °C compared with 21 °C. Decreasing the methane gas bubble diameter by decreasing the sparger pore size from 1 mm to 0.5 µm significantly improved methane removal and reactor productivity by increasing mass transfer. Premixing methane and air before sparging into the reactor saw a higher removal of methane, while sparging methane and air separately created an increase in reactor productivity. Maximum methane removal efficiency was observed to be 70.56% ± 0.54 which translated to a CH₄-EC of 93.82 ± 3.36 g CH₄ m⁻³ h⁻¹. Maximum ectoine yields was observed to be 0.579 mg ectoine L⁻¹ h⁻¹. Full article

To enhance ecosystem services (ESs) benefits and promote ecological–economic–sociologic sustainability in highly urbanized regions such as the Beijing–Tianjin–Hebei (BTH) region, it is essential to assess the dynamic changes in ESs within these regions from a functional zoning perspective and to explore the interactions between ESs. This research delved into how ESs change over space and time, using land-use projections for 2035 based on Natural Development (ND), Ecological Protection (EP), Economic Construction (EC) scenarios. This study also took a close look at the interplay of these ESs across BTH and its five distinct functional zones: the Bashang Plateau Ecological Protection Zone (BS), the Northwestern Ecological Conservation Zone (ST), the Central Core Functional Zone (HX), the Southern Functional Expansion Zone (TZ), and the Eastern Coastal Development Zone (BH). We utilize the Multiple Ecosystem Service Landscape Index (MESLI) to assess the capacity to supply multiple ESs. Key results include the following: (1) Projected land-use changes for 2035 scenarios consistently show cropland and grassland declining, while forest and urbanland expand, though the magnitude of change varies by scenario. (2) Habitat quality, carbon storage, and soil conservation displayed a “high northwest–low southeast” gradient, opposite to water yield. The average MESLI value declined in all scenarios relative to 2020, with the highest value under the EP scenario. (3) Synergies prevailed between habitat quality, carbon storage, and soil conservation, while trade-offs occurred with water yield. These relationships varied spatially—for instance, habitat quality and soil conservation were weakly synergistic in the BS but showed weak trade-offs in the HX. These insights can inform management strategies in other rapidly urbanizing regions. Full article

Hemodynamics significantly impact the biology of endothelial cells (ECs) lining the blood vessels. ECs are exposed to various hemodynamic forces, particularly frictional shear stress from flowing blood. While physiological flows are critical for the normal functioning of ECs, abnormal flow dynamics, known as disturbed flows, may trigger endothelial dysfunction leading to atherosclerosis and other vascular conditions. Such flows can occur due to sudden geometrical variations and vascular abnormalities in the cardiovascular system. In the current study, a microfluidic system was used to investigate the impact of different flow conditions (i.e, normal vs. disturbed) on ECs in vitro. We particularly explored the relationship between specific flow patterns and cellular pathways linked to oxidative stress and inflammation related to atherosclerosis. Here, we utilized a 2D cell culture perfusion system featuring an immortalized human vascular endothelial cell line (EA.hy926) connected to a modified peristaltic pump system to generate either steady laminar flows, representing healthy conditions, or disturbed oscillatory flows, representing diseased conditions. EA.hy926 were exposed to an oscillatory flow shear stress of 0.5 dynes/cm² or a laminar flow shear stress of 2 dynes/cm² up to 24 h. Following flow exposure, cells were harvested from the perfusion chamber for quantitative PCR analysis of gene expression. Reactive oxygen species (ROS) generation under various shear stress conditions was also measured using DCFDA/H2DCFDA fluorescent assays. Under oscillatory shear stress flow conditions (0.5 dynes/cm²), EA.hy926 ECs showed a 3.5-fold increase in the transcription factor nuclear factor (NFκ-B) and a remarkable 28.6-fold increase in cyclooxygenase-2 (COX-2) mRNA expression, which are both proinflammatory markers, compared to static culture. Transforming growth factor-beta (TGFβ) mRNA expression was downregulated in oscillatory and laminar flow conditions compared to the static culture. Apoptosis marker transcription factor Jun (C-Jun) mRNA expression increased in both flow conditions. Apoptosis marker C/EBP homologous protein (CHOP) mRNA levels increased significantly in oscillatory flow, with no difference in laminar flow. Endothelial nitric oxide synthase (eNOS) mRNA expression was significantly decreased in cells exposed to oscillatory flow, whereas there was no change in laminar flow. Endothelin-1 (ET-1) mRNA expression levels dropped significantly by 0.5- and 0.8-fold in cells exposed to oscillatory and laminar flow, respectively. ECs subjected to oscillatory flow exhibited a significant increase in ROS at both 4 and 24 h compared to the control and laminar flow. Laminar flow-treated cells exhibited a ROS generation pattern similar to that of static culture, but at a significantly lower level. Overall, by exposing ECs to disturbed and normal flows with varying shear stresses, significant changes in gene expression related to inflammation, endothelial function, and oxidative stress were observed. In this study, we present a practical, optimized system as an in vitro model that can be employed to investigate flow-associated diseases, such as atherosclerosis and aortic aneurysm, thereby supporting the understanding of the underlying molecular mechanisms. Full article

Kratom (Mitragyna speciosa (Korth.) Havil.) is a medicinal plant containing bioactive alkaloids, notably mitragynine and 7-hydroxymitragynine, which are psychoactive compounds with analgesic and stimulant properties. Due to safety concerns, the use of Kratom leaves and mitragynine in oral pharmaceutical products is restricted. Therefore, their potential as topical cosmeceutical agents merits further exploration. This study aimed to investigate the antioxidant and anti-adipogenic activities of Kratom ethanolic (Et-MS) and alkaloid-rich (Alk-MS) extracts, as well as purified mitragynine, to determine whether mitragynine is the major bioactive compound responsible for lipid reduction in 3T3-L1 adipocytes. The antioxidant properties were assessed using DPPH, ABTS, and FRAP assays, yielding EC₅₀ values of 0.06 mg/mL, 0.29 mg/mL, and 55 g Fe²⁺/100 g for Et-MS, respectively. In comparison, ascorbic acid (positive control) showed a DPPH EC₅₀ value of 0.002 mg/mL. Both Alk-MS and mitragynine significantly inhibited lipid accumulation in 3T3-L1 adipocytes by up to 50–70% at non-cytotoxic concentrations (≤25 µg/mL), as determined by Oil Red O staining. These findings provide preliminary in vitro evidence that phenolic constituents contribute to antioxidant capacity, while mitragynine is the principal anti-adipogenic constituent in Kratom extracts. Collectively, the results support the potential for further development of Kratom-derived extracts and mitragynine as plant-based candidates for topical or cosmeceutical applications targeting subcutaneous fat and oxidative skin damage. Full article

Waste dumpsites in developing countries are primary pollution sources impacting nearby ecosystems. This study assessed seasonal changes in soils surrounding the Hulene-B landfill (Maputo, Mozambique) and evaluated the potential for surface water contamination by leachates. A total of 71 samples were collected during the rainy and dry periods and analyzed for pH, electrical conductivity (EC), organic matter (OM), and color. The contamination potential (Pbci) was determined considering the landfill’s characteristics and local hydrological context. During the dry season, soils exhibited higher EC and OM, indicating greater retention of potentially toxic elements (PTEs) and a strong tendency for accumulation. In the rainy season, leaching processes prevailed, leading to reduced EC and OM but increased potential for contaminant mobility. The Pbci values were consistently high across both periods, confirming elevated contamination risk. Overall, the Hulene-B landfill exerts a marked influence on surrounding soils and nearby surface waters, underscoring the urgent need for structural measures to control leachate release and ash dispersion. Full article

Long-term fertilization strategies are crucial for sustainable soil health and crop productivity. However, the synergistic effect of combining straw with lime in long-term fertilization remains underexplored, particularly regarding soil micronutrient availability and tomato yield. This study examined the 10-year effects of chicken manure (M) with straw (S) and/or lime (Ca) on soil properties, micronutrient availability, and tomato yield. The results demonstrated that all of the fertilization treatments significantly altered topsoil (0–20 cm) characteristics, reducing the pH but increasing the EC and nutrient content. The combined MSCa treatment was most effective, achieving the highest levels of total carbon (19 g/kg) and tomato yield (5.6 kg/m²), which was 12–87% higher than that achieved with the other treatments. Fertilization also significantly increased the diethylenetriamine pentaacetic acid (DTPA)-extractable Fe, Mn, Cu, and Zn concentrations in both bulk soil and aggregate fractions, with availability strongly correlated with the soil total carbon and pH. The straw and lime amendments significantly improved the fruit quality by increasing the vitamin C and soluble sugar content while reducing the nitrate content. Furthermore, these treatments altered the distribution of micronutrients within the tomato organs, increasing their proportion in roots and fruits specifically. This study concludes that the integrated application of chicken manure with straw and lime is a highly effective strategy for improving soil fertility, enhancing micronutrient bioavailability, and boosting both the yield and nutritional quality of tomatoes. Full article

Honey is a natural product of high nutritional and therapeutic value, whose biological properties are closely linked to its botanical origin and chemical composition. This study aimed to characterize avocado honey in terms of botanical origin, physicochemical parameters, phenolic content, antioxidant activity, chemical profile by LC-MS/MS, and antibacterial potential. Melissopalynological analysis revealed 86.21% avocado pollen, allowing classification as monofloral honey. The sample presented amber color and a high total phenolic content (269.79 ± 1.10 mg GAE 100 g⁻¹), values higher than those commonly reported for Brazilian and international honeys. Antioxidant activity, assessed by the DPPH method, confirmed the strong radical-scavenging capacity, consistent with the phenolic profile identified (EC₅₀ 10.250 ± 0.003 mg mL⁻¹). LC-MS/MS analysis allowed the annotation of nine compounds, including caffeine, scopoletin, abscisic acid, and vomifoliol, compounds associated with antioxidant, anti-inflammatory, and metabolic regulatory activities. Although no antibacterial effect was detected against the tested oral bacterial strains, the results highlight the chemical diversity and functional potential of avocado honey. Overall, the findings reinforce the bioactive potential of avocado honey, particularly due to its strong antioxidant capacity and diversity of metabolites, supporting its value as a natural resource of nutritional and therapeutic interest. Full article

Efficient water management is essential for optimizing agricultural productivity in water-scarce regions such as the Lis Valley, Portugal. In situ measurements of soil moisture content (SMC) and electrical conductivity (EC), together with Sentinel-2-derived vegetation indices, were used to assess the crop water status and evapotranspiration dynamics of pumpkin (Cucurbita moschata ‘Butternut’) during the 2020 growing season. SMC and EC were measured at depths of 10, 20, 30, 50, and 70 cm using a TDR sensor, with strong correlations observed in the upper layers, indicating that EC can complement direct SMC measurements in characterizing near-surface moisture conditions. Sentinel-2 imagery was acquired to compute NDVI, SAVI, EVI, and GCI. In addition, NDVI values obtained from both a GreenSeeker^® sensor and Sentinel-2 imagery were compared, showing a similar temporal pattern during the season. By replacing the standard FAO-56 K_c values with those derived from each vegetation index, ET_a was recalculated to incorporate actual crop condition variability detected via satellite. ET_a estimates from RS-assisted vegetation indices agreed with those obtained using the FAO-56 method; independent ET_a measurements were not available for validation. Although such agreement is partly expected due to calibration, its confirmation for Cucurbita moschata under Mediterranean conditions—where published references are scarce—reinforces the method’s practical applicability for water management in data-limited settings. Water Productivity (WP) was estimated as 8.32 kg m⁻³, and Water Use Efficiency (WUE FAO-56) was calculated as 0.64 kg m⁻³, indicating high water use efficiency under Mediterranean smallholder irrigation conditions. These findings demonstrate that integrating high-resolution RS with continuous soil moisture monitoring can enhance precision irrigation strategies, increase crop yields, and conserve water resources in the Lis Valley. Full article

Cystic echinococcosis (CE) is an endemic zoonotic disease caused by Echinococcus granulosus, which forms cysts in ungulates’ intermediate hosts. Humans are accidental hosts, and CE affects more than one million people worldwide. Imaging remains the diagnostic gold standard, outperforming serological methods. This study presents an in silico analysis of two glyceraldehyde-3-phosphate dehydrogenase (GAPDH) isoenzymes from E. granulosus (EgGAPDH), isolated from a parasite cell line (EGPE). EgGAPDHs were recognized by sera from CE patients, identified through LC-MS/MS and PCR of metacestodes from cattle liver. One isoenzyme is intracellular (IC) (UniProt: W6UJ19), and the other is extracellular (EC) (UniProt: W6V1T8). GAPDH is involved in host–parasite interactions and metabolic processes. We characterized the physicochemical properties; linear epitopes (LEPs); and amino acid domains of EgGAPDH, its hosts, and other parasites. W6UJ19 emerged as the most promising isoenzyme as a marker of infection. Molecular dynamics simulations of isoenzymes, performed in the presence or absence of two bisphosphonates (BPs), revealed how drug binding alters conformational epitopes (CEPs) and suggested that W6UJ19 is more responsive to BP modulation. Binding affinity analysis using the MMPBSA method revealed that etidronate (EHDP) binds EgGAPDH with greater affinity than phosphate (Pi) and alendronate (AL), in the following order: EHDP > Pi > AL. Full article

The article presents a method for detecting low-intensity DDoS attacks, focused on identifying difficult-to-detect “low-and-slow” scenarios that remain undetectable by traditional defence systems. The key feature of the developed method is the statistical criteria’s (χ² and T statistics, energy ratio, reconstruction errors) integration with a combined neural network architecture, including convolutional and transformer blocks coupled with an autoencoder and a calibrated regressor. The developed neural network architecture combines mathematical validity and high sensitivity to weak anomalies with the ability to generate interpretable artefacts that are suitable for subsequent forensic analysis. The developed method implements a multi-layered process, according to which the first level statistically evaluates the flow intensity and interpacket intervals, and the second level processes features using a neural network module, generating an integral blend-score S metric. ROC-AUC and PR-AUC metrics, learning curve analysis, and the estimate of the calibration error (ECE) were used for validation. Experimental results demonstrated the superiority of the proposed method over existing approaches, as the achieved values of ROC-AUC and PR-AUC were 0.80 and 0.866, respectively, with an ECE level of 0.04, indicating a high accuracy of attack detection. The study’s contribution lies in a method combining statistical and neural network analysis development, as well as in ensuring the evidentiary value of the results through the generation of structured incident reports (PCAP slices, time windows, cryptographic hashes). The obtained results expand the toolkit for cyber-attack analysis and open up prospects for the methods’ practical application in monitoring systems and law enforcement agencies. Full article

This study assessed the effects of tetracycline (TC) on growth of Lemna aoukikusa and Spirodela polyrhiza under batch conditions. The duckweeds were exposed to a range of 0.0–5.0 mg L⁻¹ of TC for 7 days in a medium containing 10 mg L⁻¹ total nitrogen (TN) and 1 mg L⁻¹ total phosphorus (TP). The relative growth rate (RGR) of each species was determined from the frond area measurement using image analysis. The EC₅₀ values as the TC concentrations causing a 50% reduction in RGR, were 4.4 mg L⁻¹ for L. aoukikusa and 0.65 mg L⁻¹ for S. polyrhiza. At 5.0 mg L⁻¹ TC, TP removal decreased to 60% in the L. aoukikusa culture and 77% in the S. polyrhiza culture, compared to 85–91% and 96%, respectively, under lower TC exposure. Nevertheless, TN and TP removals were not significantly impaired at TC concentrations found in swine wastewater. The TC removals were 76–94% for the L. aoukikusa culture and 68–91% for the S. polyrhiza culture, which were attributed to adsorption and plant uptake. These findings highlight the feasibility of duckweed-based stabilization ponds for simultaneous antibiotic attenuation and nutrient removal. Full article

Soil toxicity resulting from either natural or anthropogenic heavy metal contamination was evaluated through a nitrifying bacteria bioassay focused on the inhibition of oxygen consumption. Every contaminated soil sample inhibited the nitrifying bacteria bioassay, with inhibition levels ranging from 71% to 100%. The optimal conditions for maximizing O₂ consumption during the test procedure were established as follows: a test culture volume of 1 mL, a soil sample weight of 1 g, a rotation rate of 100 revolutions per minute, and a reaction duration of 48 h. In low- or uncontaminated soils, oxygen consumption ranged from 3.2 mL to 3.0 mL from a headspace volume of 1 mL filled with O₂. In contrast, contaminated soils exhibited a lower range, with values between 0.1 mL and 1.0 mL. EC50 levels for NB O₂ consumption were: Cr⁶⁺ 1.21 mg/kg; Cu²⁺ 6.92 mg/kg; Ag⁺ 8.38 mg/kg; As³⁺ 8.99 mg/kg; Ni²⁺ 10.35 mg/kg; Hg²⁺ 11.01 mg/kg; Cd²⁺ 31.33 mg/kg; Pb²⁺ 129.62 mg/kg. Values for inherent test variability (CVi), variation resulting from the natural characteristics of soil (CVns), and minimal detectable difference (MDD) were found to range between 1.6% and 4.7%, 7.8% and 14.6%, and 2.9% and 5.9%, respectively. A 10% toxicity threshold was set as the maximal tolerable inhibition (MTI) for effective soil toxicity assessment. Nitrifying bacteria bioassays offer a fast, affordable, and user-friendly tool for real-time soil toxicity assessment, boosting soil health monitoring and ecosystem protection. Full article

Background/Objectives: Despite advances in targeted therapies, a substantial proportion of high-risk endometrial carcinomas (EC) do not respond to treatment and have a poor prognosis. The identification of prognostic and predictive biomarkers to improve patient stratification is therefore a clinical priority. L1 cell adhesion molecule (L1CAM) is a promising biomarker in EC; however, its soluble circulating form (sL1CAM) has been poorly investigated. This study aimed to evaluate the prognostic and predictive significance of sL1CAM in high-risk ECs. Methods: High-risk EC patients, treated with surgery and platinum-based adjuvant chemotherapy, were retrospectively enrolled. sL1CAM levels were quantified in 72 preoperative serum samples by enzyme-linked immunosorbent assay (ELISA). Results: High sL1CAM levels were associated with advanced age and non-endometrioid histology. Across the entire patient cohort, higher sL1CAM concentrations significantly correlated with worse prognosis in terms of DSS and PFS in univariate (DSS: HR = 2.22, p = 0.028; PFS: HR = 1.21, p = 0.041) and multivariate (DSS: HR = 2.13, p = 0.041; PFS: HR = 1.93, p = 0.048) analyses. Stratification by histological type revealed a significant prognostic association only in the endometrioid subgroup, both in univariate and multivariate analyses. Moreover, in this subgroup, elevated sL1CAM levels were associated with shorter time to recurrence after chemotherapy, both in univariate (PFI: HR = 2.69, p = 0.027) and multivariate (PFI: HR = 2.97, p = 0.017) analysis, and significantly predicted relapse within 6 months (OR = 7.83, p = 0.027). Conclusions: sL1CAM is associated with poor prognosis in high-risk EC and seems to be associated with platinum response in endometrioid tumors. These findings support its potential role as a biomarker to improve risk stratification, warranting validation in larger, prospective studies. Full article

Grape pomace (GP), a by-product of winemaking, is rich in polyphenols and fiber, making it a promising and sustainable feed supplement for ruminants. This study evaluated the safety and productive impact of a 5% GP-supplemented diet (GP5) including non-lactating end-cycle (EC) ewes regularly destined for slaughter and human consumption, and lactating (LAC) ewes, over a 30-day period. Control (CTRL) animals received a standard pellet diet with no GP inclusion. Sampling was performed at four time points (T0, T10, T20, and T30), corresponding to days 0, 10, 20, and 30 of the experimental period. The study assessed clinical status, hematology/biochemistry (T0 and T30), milk composition (T0, T10, T20, and T30), meat quality traits and oxidative stability in EC ewes (T30). Since no significant differences were observed in the CTRL animals, the effects were evaluated within the GP5 group by comparing T0 vs. T30. Meat quality was assessed by comparing EC-GP5 to CTRL at T30. The GP extract showed a high total phenolic content (254.02 ± 20.39 mg GAE/g DW). No clinical or hematological alterations were observed, and most values remained within physiological ranges. Biochemical analysis revealed significant increases in albumin, bilirubin, creatinine, and triglycerides (p < 0.05), with significant decreases in plasma urea and glucose (p < 0.05). In LAC-GP5 ewes, milk urea and lactose concentrations decreased (p < 0.05), while pH increased (p < 0.05), with no significant changes in fat or casein content. These findings are consistent with reduced ruminal propionate availability, leading to decreased hepatic gluconeogenesis and lactose synthesis, with secondary effects on nitrogen metabolism and the acid–base profile of milk. In EC-GP5 ewes, meat quality traits were unaffected, and DPPH scavenging activity did not differ from CTRL (p > 0.05). GP5 was metabolically safe, induced adaptive changes in milk composition, and had no negative effects on meat quality, supporting the valorization of grape pomace as a sustainable feed resource. This trial was designed as a metabolic safety assessment, representing a preliminary step toward future mechanistic and molecular investigations. Full article