Search Results (1,487)

While the Ankom RF system facilitates efficient high-throughput in vitro fermentation studies, its high cost and limited flexibility constrain its broader applicability. To address these limitations, we developed and validated a low-cost, modular gas monitoring system (FerME), assembled from commercially available components. To evaluate its performance and reproducibility relative to the Ankom RF system (Ankom Technology, Macedon, NY, USA), in vitro rumen fermentation experiments were conducted under strictly controlled and identical conditions. Whole rumen contents were collected approximately 2 h post-feeding from individual mid- or late-lactation dairy cows and immediately transported to the laboratory. Each fermenter received 50 mL of processed rumen fluid, 100 mL of anaerobically prepared artificial saliva buffer, and 1.2 g of the donor cow’s diet. Bottles were sealed with the respective system’s pressure sensors, flushed with CO₂, and incubated in a 50 L water bath maintained at 39 °C. FerME (New Taipei City, Taiwan) and Ankom RF fermenters were placed side-by-side to ensure uniform thermal conditions. To assess the effect of filter bag use, an additional trial employed Ankom F57 filter bags (Ankom Technology, Macedon, NY, USA; 25 μm pore size). Trial 1 revealed no significant differences in cumulative gas production, volatile fatty acids (VFAs), NH₃-N, or pH between systems (p > 0.05). However, the use of filter bags reduced gas output and increased propionate concentrations (p < 0.05). Trial 2, which employed filter bags in both systems, confirmed comparable results, with the FerME system demonstrating improved precision (CV: 4.8% vs. 13.2%). Gas composition (CH₄ + CO₂: 76–82%) and fermentation parameters remained consistent across systems (p > 0.05). Importantly, with 12 pressure sensors, the total cost of FerME was about half that of the Ankom RF system. Collectively, these findings demonstrate that FerME is a reliable, low-cost alternative for real-time rumen fermentation monitoring and could be suitable for studies in animal nutrition, methane mitigation, and related applications. Full article

Urban forests are vital to cities because they provide a range of ecosystem services, including carbon (C) sequestration, air purification, and urban cooling. However, urban forestry also generates significant amounts of organic waste, such as grass clippings, pruned tree branches, and fallen tree leaves and woody debris that can contribute to greenhouse gas (GHG) emissions if not properly managed. In this study, we investigated the effect of wheat straw biochar (produced at 500 °C) on GHG emissions from two types of urban forestry waste: green waste (GW) and yard waste (YW), using a 100-day laboratory incubation experiment. Overall, GW released more CO₂ than YW, but biochar addition reduced cumulative CO₂ emissions by 9.8% in GW and by 17.6% in YW. However, biochar increased CH₄ emissions from GW and reduced the CH₄ sink strength of YW. Biochar also had contrasting effects on N₂O emissions, increasing them by 94.3% in GW but decreasing them by 61.4% in YW. Consequently, the highest global warming potential was observed in biochar-amended GW (125.3 g CO₂-eq kg⁻¹). Our findings emphasize that the effect of biochar on GHG emissions varies with waste type and suggest that selecting appropriate biochar types is critical for mitigating GHG emissions from urban forestry waste. Full article

Autologous therapies are currently being studied to determine if they can modulate the course of knee osteoarthritis symptoms and/or disease progression. One potential therapeutic target is the polarization of pro-inflammatory M1 macrophages to pro-healing M2 macrophages. The autologous therapy, Autologous Protein Solution (APS), was incubated with donor-matched human peripheral-derived macrophages for 10 days. M1 pro-inflammatory macrophages were determined by the percentage of CD80+ and M2 pro-healing macrophages were determined by CD68+ and CD163+ by epifluorescent microscopy. To determine clinical effectiveness, an APS-specific minimal clinically important improvement (MCII) using an anchor-based method was calculated in a randomized controlled trial of APS (n = 46) and then applied to a real-world registry study (n = 78) to determine the percentage of pain responders. Compared to control media, APS statistically increased the percentage of M2 macrophages and decreased the percentage of M1 macrophages, while platelet-poor plasma had no effect on polarization. In the randomized controlled trial (RCT), the MCII at the 12-month follow-up visit was calculated as 2.0 points on the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) pain scale and 7.5 points on the WOMAC function scale. Applying this MCII to the real-world registry data, 62.5% of patients met the MCII with an average of 4.7 ± 2.5 points of improvement in pain. Autologous therapies can influence macrophage polarization and have demonstrated clinical effectiveness in a real-world patient setting. Full article

Concern over the presence of pharmaceutical waste in the environment has prompted research into the management of emerging organic micropollutants (EOMs). In response, sustainable technologies have been applied as alternatives to reduce the effects of these contaminants. This study investigated the capacity of filamentous fungi isolated from iron mine soil in the Amazon region to biodegrade the drug chloroquine diphosphate. An initial screening assessed the growth of four fungal strains on solid media containing chloroquine diphosphate: Trichoderma pseudoasperelloides CBMAI 2752, Penicillium rolfsii CBMAI 2753, Talaromyces verruculosus CBMAI 2754, and Penicillium sp. cf. guaibinense CBMAI 2758. Among them, Penicillium sp. cf. guaibinense CBMAI 2758 was selected for further testing in liquid media. A Box–Behnken factorial design was applied with three variables, pH (5, 7, and 9), incubation time (5, 10, and 15 days), and chloroquine diphosphate concentration (50, 75, and 100 mg·L⁻¹), totaling 15 experiments. The samples were analyzed by gas chromatography–mass spectrometry (GC-MS). The most effective conditions for chloroquine biodegradation were pH 7, 100 mg·L⁻¹ concentration, and 10 days of incubation. Four metabolites were identified: one resulting from N-deethylation M1 (N4-(7-chloroquinolin-4-yl)-N1-ethylpentane-1,4-diamine), two from carbon–carbon bond cleavage M2 (7-chloro-N-ethylquinolin-4-amine) and M3 (N1,N1-diethylpentane-1,4-diamine), and one from aromatic deamination M4 (N1-ethylbutane-1,4-diamine) by enzymatic reactions. The toxicity analysis showed that the products obtained from the biodegradation of chloroquine were less toxic than the commercial formulation of this compound. These findings highlight the biotechnological potential of Amazonian fungi for drug biodegradation and decontamination. Full article

The barbel chub (Squaliobarbus curriculus) exhibits remarkable resistance to grass carp reovirus (GCRV), a devastating pathogen in aquaculture. To reveal the molecular basis of this resistance, we investigated complement factor D (DF)—a rate-limiting serine protease governing alternative complement pathway activation. Molecular cloning revealed that the barbel chub DF (ScDF) gene encodes a 1251-bp cDNA sequence translating into a 250-amino acid protein. Crucially, bioinformatic characterization identified a unique N-glycosylation site at Asn¹³⁹ in ScDF, representing a structural divergence absent in grass carp (Ctenopharyngodon idella) DF (CiDF). While retaining a conserved Tryp_SPc domain harboring the catalytic triad (His⁶¹, Asp¹⁰⁹, and Ser²⁰⁴) and substrate-binding residues (Asp¹⁹⁸, Ser²¹⁹, and Gly²²¹), sequence and phylogenetic analyses confirmed ScDF’s evolutionary conservation, displaying 94.4% amino acid identity with CiDF and clustering within the Cyprinidae. Expression profiling revealed constitutive ScDF dominance in the liver, and secondary prominence was observed in the heart. Upon GCRV challenge in S. curriculus kidney (SCK) cells, ScDF transcription surged to a 438-fold increase versus uninfected controls at 6 h post-infection (hpi; p < 0.001)—significantly preceding the 168-hpi response peak documented for CiDF in grass carp. Functional validation showed that ScDF overexpression suppressed key viral capsid genes (VP2, VP5, and VP7) and upregulated the interferon regulator IRF9. Moreover, recombinant ScDF protein incubation induced interferon pathway genes and complement C3 expression. Collectively, ScDF’s rapid early induction (peaking at 6 hpi) and multi-pathway coordination may contribute to barbel chub’s GCRV resistance. These findings may provide molecular insights into the barbel chub’s high GCRV resistance compared to grass carp and novel perspectives for anti-GCRV breeding strategies in fish. Full article

Background/Objectives: Tamoxifen, a selective estrogen receptor modulator widely used as an adjunct in the treatment of breast cancer, has known effects on bone metabolism, although its impact on osseointegration and cellular responses during early bone healing remains unclear. Understanding these effects is essential given the increasing use of dental implants in cancer survivors. The study aimed to observe the influence of tamoxifen on human osteosarcoma (SAOS-2) cells lines, as well on the osseointegration of titanium implants in ovariectomized female rats. Methods: SAOS-2 cells were incubated with Dulbecco’s modified growth medium. Six titanium (Ti) disks were used at each time point. The samples were divided into groups with the presence (TAM, n = 36) or not (CTR, n = 36) of tamoxifen in a concentration of 2 μM. In vivo, 72 animals were divided in groups with bilateral ovariectomy or SHAM and tamoxifen administration or not (15 mg/kg). Cell viability, mineralization rate, and collagen synthesis were assessed, as well as bone/implant contact (BIC) and bone ingrowth (BIN). Results: Tamoxifen caused a decrease in SAOS-2 viability, although an increase in the mineralization rate was observed. In vivo, the TAM groups presented higher BIC and BIN when compared to their control, but a lower percentage of mature collagen cells. Conclusions: Based on our findings, in vitro, the therapy with TAM slightly reduced the viability of SAOS-2 cells while significantly increasing the mineralization rate. In vivo, the therapy positively influenced BIC and BIN during the osseointegration phase. Full article

The aim of this study was to analyze how recombinant rabbit NGF (Nerve Growth Factor) encapsulated in chitosan (rrβNGFch) affects sperm viability, motility, capacitation, acrosome reaction (AR), kinetic traits, and apoptosis after 30 min and 2 h of storage. Specific intracellular signaling pathways associated with either cell survival, such as protein kinase B (AKT) and extracellular signal-regulated kinases 1/2 (ERK1/2), or programmed cell death, such as c-Jun N-terminal kinase (JNK), were also analyzed. The results confirmed the effect of rrβNGFch on capacitation and AR, whereas a longer storage time (2 h) decreased all qualitative sperm traits. AKT and JNK did not show treatment-dependent activation and lacked a correlation with functional traits, as shown by ERK1/2. These findings suggest that rrβNGFch may promote the functional activation of sperm cells, particularly during early incubation. The increase in capacitation and AR was not linked to significant changes in pathways related to cell survival or death, indicating a specific action of the treatment. In contrast, prolonged storage negatively affected all sperm parameters. ERK1/2 activation correlated with capacitation, AR, and apoptosis, supporting its role as an NGF downstream mediator. Further studies should analyze other molecular mechanisms of sperm and the potential applications of NGF in assisted reproduction. Full article

Objective: To investigate the neuroprotective effects of a Rho-associated kinase (ROCK) inhibitor on retinal ganglion cells (RGCs) in vitro and in vivo. Methods: For in vivo studies, a unilateral optic nerve crush mouse model was established. Then, 100 mM Y-27632 (a ROCK inhibitor) or saline was applied to the experimental eyes once a day for 14 days. The effects of the ROCK inhibitor were evaluated by counting the surviving RGCs in the enucleated flat retina tissues and measuring the inner retinal thickness using optical coherence tomography (OCT), the amplitude of the electroretinogram (ERG), and the change in intraocular pressure (IOP). For the in vitro study, RGCs were isolated from five-day-old mice using a modified immunopanning method with magnetic beads. The isolated RGCs were incubated for 72 h with various concentrations of Y-27632, after which TUNEL assays were performed to determine the number of surviving RGCs. Results: Y-27632 has neuroprotective effects, as it significantly increased the number of surviving RGCs by approximately 6.3%. OCT and ERG data also revealed that Y-27632 induced neuroprotective effects in vivo; furthermore, Y-27632 reduced IOP by approximately 18.3%. The in vitro study revealed the dose-dependent neuroprotective effects of Y-27632, with the highest dose of Y-27632 (1000 nM) increasing the RGC survival rate after 72 h of incubation compared with that of the control. Conclusions: The ROCK inhibitor Y-27632 may exert some neuroprotective effects on RGCs when it is used as an eye drop through an IOP-independent mechanism. 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

Background and aim: ANGPTL3 is a hepatokine acting as a negative regulator of lipoprotein lipase (LPL) through its N-terminal domain. Besides this activity, the C-terminal domain of ANGPTL3 interacts with integrin αVβ3. Since integrins are involved in inflammation and in the initiation of atherosclerotic plaque, the aim of our study was to evaluate the potential direct pro-inflammatory action of ANGPTL3 through the interaction of the fibrinogen-like domain and integrin αVβ3. Methods: We utilized cultured THP-1 human-derived macrophages and evaluated their pro-inflammatory phenotype in response to treatment with human recombinant ANGPTL3 (hANGPTL3). By Western blot, RT-qPCR, biochemical analysis, and ELISA assays, we determined the expression of genes and proteins involved in lipid metabolism and inflammatory response as well as intracellular cholesterol and triglyceride levels. In addition, we evaluated the effect of hANGPTL3 on the cellular cholesterol efflux process. Results: Incubation of THP-1-derived macrophages with 100 ng/mL of hANGPTL3 increased the mRNA expression of the pro-inflammatory cytokines IL-1β, IL-6, and TNFα (respectively, 1.87 ± 0.08-fold, 1.35 ± 0.11-fold, and 2.49 ± 0.43-fold vs. control). The secretion of TNFα, determined by an ELISA assay, was also induced by hANGPTL3 (1.98 ± 0.4-fold vs. control). The pro-inflammatory effect of hANGPTL3 was partially counteracted by co-treatment with the integrin αVβ3 inhibitor RGD peptide, reducing the mRNA levels of IL-1β (3.35 ± 0.35-fold vs. 2.54 ± 0.25-fold for hANGPTL3 vs. hANGPTL3 + RGD, respectively). Moreover, hANGPTL3 reduced cholesterol efflux to apoA-I, with a parallel increase in the intracellular triglyceride and cholesterol contents by 31.2 ± 2.8% and 20.0 ± 4.1%, respectively, compared to the control. Conclusions: ANGPTL3 is an important liver-derived regulator of plasma lipoprotein metabolism, and overall, our results add a new important pro-inflammatory activity of this circulating protein. This new function of ANGPTL3 could also be related to triglyceride and cholesterol accumulation into macrophages. 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

Pyrochar has been identified as a favorable soil conditioner that can effectively ameliorate soil acidification. Hydrochar is considered a more affordable carbon material than pyrochar, but its effect on the process of soil acidification has yet to be investigated. An indoor incubation and a soil column experiment were conducted to study the effect of rice straw hydrochar application on nitrification and NO₃⁻-N leaching in acidic red soil. Compared to the control and pyrochar treatments, respectively, hydrochar addition mitigated the net nitrification rate by 3.75–48.75% and 57.92–78.19%, in the early stage of urea fertilization. This occurred mainly because a greater amount of dissolved organic carbon (DOC) was released from hydrochar than the other treatments, which stimulated microbial nitrogen immobilization. The abundances of ammonia-oxidizing archaea and ammonia-oxidizing bacteria were dramatically elevated by 25.62–153.19% and 12.38–22.39%, respectively, in the hydrochar treatments because of DOC-driven stimulation. The cumulative leaching loss of NO₃⁻-N in soils amended with hydrochar was markedly reduced by 43.78–59.91% and 61.70–72.82% compared with that in the control and pyrochar treatments, respectively, because hydrochar promoted the soil water holding capacity by 2.70–9.04% and reduced the residual NO₃⁻-N content. Hydrochar application can dramatically diminish total H⁺ production from soil nitrification and NO₃⁻-N leaching. Thus, it could be considered an economical soil amendment for ameliorating soil acidification. Full article

Background/Objectives: The emergence of cathinone-based psychostimulants necessitates ongoing research and analysis of the characteristics and properties of novel derivatives. The metabolic fate of five novel cathinone-derived substances (ASProp, MASProp, MASPent, PySProp, and PySPent) containing a selenophene moiety was investigated in vitro and in vivo. Methods: All compounds were incubated individually with pooled human liver S9 fraction. A monooxygenase activity screening investigating the metabolic contribution of eleven recombinant phase I isoenzymes was conducted. Rat urine after oral administration was prepared by urine precipitation. Liquid chromatography–high-resolution tandem mass spectrometry was used for the analysis of all samples. Reversed-phase liquid chromatography (RPLC) and zwitterionic hydrophilic interaction liquid chromatography (HILIC) were used to evaluate and compare the metabolites’ chromatographic resolution. Results: Phase I reactions of ASProp, MASProp, MASPent, PySProp, and PySPent included N-dealkylation, hydroxylation, reduction, and combinations thereof. The monooxygenase activity screening revealed the contribution of various isozymes. Phase II reactions detected in vivo included N-acetylation and glucuronidation. Both chromatographic columns complemented each other. Conclusions: All substances revealed metabolic reactions comparable to those observed for other synthetic cathinones. Contributions from isozymes to their metabolism minimized the risk of drug–drug interactions. The identified metabolites should be considered as targets in human biosamples, especially in urine screening procedures. RPLC and HILIC can both be recommended for this purpose. Full article

Ruminant livestock contribute significantly to methane emissions, necessitating sustainable mitigation strategies. Essential oils (EOs) show promise for modulating ruminal fermentation, but their synergistic effects remain underexplored. Two 24 h in vitro experiments evaluated the synergistic effects of EO blends on rumen microbial fermentation. Exp. 1 screened five oils using two triad combinations. Triad 1 tested 10 combinations of thyme (THY), peppermint (PPM), and cinnamon leaf (CIN) oils. Triad 2 tested 10 combinations of anise (ANI), clove leaf (CLO), and peppermint (PPM) oils. Each blend was tested at 400 mg/L, using batch culture methods measuring: pH, ammonia-N (NH₃-N), and volatile fatty acids (VFAs). The two most effective blends, designated as T1 and T2, were selected for Exp. 2 to assess total gas and methane (CH₄) production using pressure transducer methods. All treatments were incubated in a rumen fluid–buffer mix with a 50:50 forage-to-concentrate substrate (pH 6.6). In Exp. 1, data were analyzed according to the Simplex Centroid Design using R-Studio. In Exp. 2, an analysis was conducted using the MIXED procedure in SAS. Mean comparisons were assessed through Tukey’s test. The results from Exp. 1 identified CIN+PPM (80:20) and ANI+CLO (80:20) as optimal combinations, both increasing total VFAs while reducing acetate/propionate ratios and NH₃-N concentrations. In Exp. 2, both combinations significantly reduced total gas and CH₄ productions compared to the control, with CIN+PPM achieving the greatest methane reduction (similar to monensin, the positive control). Specific essential oil combinations demonstrated synergistic effects in modulating rumen fermentation and reducing methane emissions, offering potential for sustainable livestock production. Further in vivo validation is required to optimize dosing and assess long-term effects on animal performance. Full article

Urine-derived and plant-derived benzoic acid can accumulate within soil, and it likely changes the soil microbial community as well as N₂O emissions; however, its mechanism is not clear. This study conducted an incubation experiment to monitor N₂O emissions under low moisture (40% water-filled pore space (WFPS)) and high moisture (85% WFPS) conditions. Metagenomic sequencing and q-PCR methods were used to determine the link between N₂O emissions and the composition and functions of soil microbiota. Benzoic acid (BA) was found to significantly, yet dose-dependently, impact N₂O emissions; that is, low BA concentrations increased N₂O, whereas high BA decreased N₂O. However, this was only found under high moisture conditions. In contrast, BA had little impact on N₂O emissions under low moisture conditions. Under high moisture conditions, BA increased the gene copy number of bacteria and fungi, and decreased the ratio of bacteria to fungi. Similarly, BA significantly increased the abundance of denitrification functional genes, but reduced the (NirS + NirK)-to-NosZ ratio at the peak of emission. This is in agreement with the observation of the increased relative abundance of genes encoding N₂O reductase (EC 1.7.2.4) and NO₃⁻ heterotrophic reductase (EC 1.7.1.15, EC 1.7.2.2) in the metagenomic analysis. In summary, high concentrations of benzoic acid reduce N₂O emissions by promoting the reduction of N₂O. This study revealed the impact of BA on soil microbiota and highlighted the favorable conditions and underlying mechanism behind BA’s significant impact on soil N₂O emissions. This study’s novelty lies in the fact that it deepens our understanding of the complicated role of root exudates and metabolites of animals and plants in soil. Full article