Search Results (808)

Acetate may act as a signaling molecule, regulating Paracin 1.7 production via quorum sensing (QS) in Lacticaseibacillus paracasei HD1.7. The “acetate switch” phenomenon requires mechanistic exploration to optimize Paracin 1.7 production. The “acetate switch” phenomenon delays with higher glucose levels (30 h, 36 h, and 96 h). Before the occurrence of the “acetate switch”, the ATP content increases and peaks at the “acetate switch” point and the NAD⁺/NADH ratio decreases, indicating energy changes. Moreover, the QS genes used for the pre-regulation of bacteriocin, such as prcKR, comCDE, were highly expressed. After the “acetate switch”, the ATP content decreased and the QS genes for the post-regulation of bacteriocin were highly expressed, such as rggs234 and sigma70-1/70-2. The “acetate switch” could act as an energy switch, regulating bacterial growth and QS genes. Before and after the “acetate switch”, some metabolic pathways were significantly altered according to the transcriptomic analysis by HD1.7 and HD1.7-Δpta. In this study, acetate was used as an input signal to regulate the two-component system, significantly influencing the bacteriocin expression system. And this study clarifies the roles of acetate, energy, and quorum sensing in promoting Paracin 1.7 production, providing a theoretical basis for optimizing the bacteriocin fermentation process of HD1.7. Full article

The process of aging in organisms is associated with progressive metabolic changes that affect energy production. In our study, we compared the activities/concentrations of components related to the Krebs cycle and the respiratory chain (such as acetyl-CoA, IDH, AKG, succinate, fumarate, NADH₂, UQCR, COX and ATP) in the hemolymph and fat body segments (tergites 3 and 5, sternite) in naturally and prematurely (affected by V. destructor) aging workers. Tergite 3 showed the highest metabolic activity, indicating its key role in energy storage and production. In naturally aging workers, the concentrations/activities of the tested components were higher in all the segments of the fat body and all the age groups when compared to the prematurely aging workers. The concentrations/activities of these components increased with age, usually reaching the maximum at 28 days of age in the fat body segments of naturally aging workers, and then decreasing in the oldest ones (at 35 days of age). An analysis of changes in the metabolic processes can provide a lot of important information on the mechanisms of aging. In the future, such studies can contribute to the development of effective strategies to delay the aging processes and improve the overall condition of bee colonies. Full article

China’s southwestern karst landscapes support remarkable cavefish diversity, especially within Sinocyclocheilus, the world’s largest cavefish genus. Using integrative taxonomic methods, we describe Sinocyclocheilus wanlanensis sp. nov., found in a subterranean river in Guizhou Province. This species lacks horn-like cranial structures; its eyes are either reduced to a dark spot or absent. It possesses a pronounced nuchal hump and a forward-protruding, duckbill-shaped head. Morphometric analysis of 28 individuals from six species shows clear separation from related taxa. Nano-CT imaging reveals distinct vertebral and cranial features. Phylogenetic analyses of mitochondrial cytb and ND4 genes place S. wanlanensis within S. angularis group as sister to S. bicornutus, with p-distances of 1.7% (cytb) and 0.7% (ND4), consistent with sister-species patterns within the genus. Sinocyclocheilus wanlanensis is differentiated from S. bicornutus by its eyeless or degenerate-eye condition and lack of bifurcated horns. It differs from S. zhenfengensis, its morphologically closest species, in having degenerate or absent eyes, shorter maxillary barbels, and pelvic fins that reach the anus. The combination of morphological and molecular evidence supports its recognition as a distinct species. Accurate documentation of such endemic and narrowly distributed taxa is important for conservation and for understanding speciation in cave habitats. Full article

As distant hybridization has profound implications for evolutionary biology, aquaculture, and biodiversity conservation, this study aims to elucidate patterns of maternal inheritance, genetic divergence, and phylogenetic relationships by synthesizing mitochondrial genome (mitogenome) data from 74 distant hybrid fish species. These hybrids span diverse taxa, including 48 freshwater and 26 marine species, with a focus on Cyprinidae (n = 35) and Epinephelus (n = 14), representing the most frequently hybridized groups in freshwater and marine systems, respectively. Mitogenome lengths were highly conserved (15,973 to 17,114 bp); however, the genetic distances between hybrids and maternal species varied from 0.001 to 0.17, with 19 hybrids (25.7%) showing distances >0.02. Variable sites in these hybrids were randomly distributed but enriched in hypervariable regions, such as the D-loop and NADH dehydrogenase subunits 1, 3 and 6 (ND2, ND3, and ND6) genes, likely reflecting maternal inheritance (reported in Cyprinus carpio × Carassius auratus). Moreover, these genes were under purifying selection pressure, revealing their conserved nature. Phylogenetic reconstruction using complete mitogenomes revealed three distinct clades in hybrids: (1) Acipenseriformes, (2) a freshwater cluster dominated by Cypriniformes and Siluriformes, and (3) a marine cluster comprising Centrarchiformes, Pleuronectiformes, Scombriformes, Cichliformes, Anabantiformes, Tetraodontiformes, Perciformes, and Salmoniformes. The prevalence of Cyprinidae hybrids underscores their importance in aquaculture for hybridization, where traits such as rapid growth and disease resistance are enhanced. In contrast, marine hybrids are valued for their market value and adaptability. While mitogenome data robustly support maternal inheritance in most cases, exceptions suggest complex mechanisms, such as doubly uniparental inheritance (DUI), in distantly related crosses. Moreover, AT-skew of genes in hybrids revealed a paternal leakage of traits in mitogenomes. This study also highlights ecological risks, such as genetic swamping in native populations, emphasizing the need for responsible hybridization practices. These findings advance our understanding of the role of hybridization in fish evolution and aquaculture, providing a genomic framework and policy recommendations for optimizing breeding programs, hybrid introduction, and mitigating conservation challenges. Full article

Background/Objectives: Oxidative stress constitutes a principal pathophysiological mechanism driving neurodegeneration and brain aging. α-Ketoglutarate (AKG), a key intermediate of the tricarboxylic acid (TCA) cycle, has shown potential in longevity and oxidative stress resistance. However, the role of AKG in oxidative stress-induced neuronal senescence and its interaction with the mTOR signaling pathway during neuronal aging remain poorly understood, posing a key challenge for developing senescence-targeted therapies. Methods: We investigated the neuroprotective effects of AKG using H₂O₂-induced senescence in HT22 cells and a D-galactose-induced brain aging mouse model. Assessments encompassed SA-β-gal staining, EdU incorporation, mitochondrial membrane potential (JC-1), and ROS measurement. Antioxidant markers, ATP levels, and the NAD⁺/NADH ratio were also analyzed. Proteomic profiling (DIA-MS) and KEGG/GSEA enrichment analyses were employed to identify AKG-responsive signaling pathways, and Western blotting validated changes in mTOR signaling and downstream effectors. Results: AKG significantly alleviated H₂O₂-induced senescence in HT22 cells, evidenced by enhanced cell viability, reduced ROS level, restored mitochondrial function, and downregulated p53/p21 expression. In vivo, AKG administration improved cognitive deficits and vestibulomotor dysfunction while ameliorating brain oxidative damage in aging mice. Proteomics revealed mTOR signaling pathways as key targets for AKG’s anti-aging activity. Mechanistically, AKG suppressed mTOR phosphorylation and activated ULK1, suggesting modulation of autophagy and metabolic homeostasis. These effects were accompanied by enhanced antioxidant enzyme activities and improved redox homeostasis. Conclusions: Our study demonstrates that AKG mitigates oxidative stress-induced neuronal senescence through suppression of the mTOR pathway and enhancement of mitochondrial and antioxidant function. These findings highlight AKG as a metabolic intervention candidate for age-related neurodegenerative diseases. Full article

The production of β-alanine from fatty acid feedstocks presents a promising synthetic strategy due to its high carbon yield. However, the excessive reducing power generated during fatty acid utilization disrupts cellular redox balance, adversely affecting metabolism and limiting the efficiency and final yield of β-alanine production. To address this challenge, we engineered a co-production system in which excess reducing equivalents generated during fatty acid β-oxidation and β-alanine biosynthesis were consumed by growth-coupled lycopene biosynthesis. The resulting dual-pathway strain, SA01, achieved 44.78 g/L β-alanine and 3.07 g/L lycopene in bioreactor fermentation, representing a 21.45% increase in β-alanine production compared to the β-alanine-producing strain WA01, and a 74.43% increase in lycopene production compared to the lycopene-producing strain LA01. Further optimization in strain SA06, involving cofactor engineering to shift redox flow from NADH to NADPH, enhanced the titers to 52.78 g/L β-alanine and 3.61 g/L lycopene. Metabolite analysis confirmed a decrease in intracellular NADH and FADH₂ levels in SA06, indicating restoration of redox balance during the late fermentation phase. Additional improvements in the fermentation process, including gradual carbon source switching, optimization of the induction strategy, and fine-tuning of conditions during both growth and bioconversion phases, resulted in further increases in product titers, reaching 72 g/L β-alanine and 6.15 g/L lycopene. This study offers valuable insights into the development of microbial co-production systems, highlighting the critical role of dynamic cofactor and redox balance management, as well as process optimization, in improving production efficiency. Full article

Lactate dehydrogenase (LDH) catalyzes the reversible interconversion of pyruvate and lactate, coupled with the redox cycling of NADH and NAD⁺. While LDHA has been extensively studied as a therapeutic target, particularly in cancer, due to its role in the Warburg effect, LDHB remains underexplored, despite its involvement in the metabolic reprogramming of specific cancer types, including breast and lung cancers. Most known LDH inhibitors are designed against the LDHA isoform and act competitively at the active site. In contrast, LDHB exhibits distinct kinetic properties, substrate preferences, and structural features, warranting isoform-specific screening strategies. In this study, 115 natural compounds previously reported as LDHA inhibitors were systematically evaluated for LDHB inhibition using an integrated in silico and in vitro approach. Virtual screening identified 16 lead phytochemicals, among which luteolin and quercetin exhibited uncompetitive inhibition of LDHB, as demonstrated by enzyme kinetic assays. These findings were strongly supported by molecular docking analyses, which revealed that both compounds bind at an allosteric site located at the dimer interface, closely resembling the binding mode of the established LDHB uncompetitive inhibitor AXKO-0046. In contrast, comparative docking against LDHA confirmed their active-site binding and competitive inhibition, underscoring their isoform-specific behavior. Our findings highlight the necessity of assay conditions tailored to LDHB’s physiological role and demonstrate the application of a previously validated colorimetric assay for high-throughput screening. This work lays the foundation for the rational design of selective LDHB inhibitors from natural product libraries. Full article

The mevalonate pathway is crucial for synthesizing isopentenyl pyrophosphate (IPP), the universal precursor of terpenoids, with 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) serving as the rate-determining enzyme that catalyzes the reduction of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) to mevalonate, requiring NAD(P)H as an electron donor. Improving the cofactor promiscuity of HMGR can facilitate substrate utilization and terpenoid production by overcoming cofactor specificity limitations. In this study, we heterologously expressed rpHMGR from Ruegeria pomeroyi in Escherichia coli BL21(DE3) for the first time and established that it predominantly utilizes NADH. To broaden its cofactor usage, we employed Molecular Operating Environment (MOE)-assisted design to engineer the cofactor binding site, creating a dual-cofactor-utilizing mutant, D154K (the substitution of aspartic acid with lysine at residue 154). This mutant exhibited a significant 53.7-fold increase in activity toward NADPH, without compromising protein stability at physiological temperatures. The D154K mutant displayed an optimal pH of 6, maintaining over 80% of its catalytic activity across the pH range of 6–8, regardless of whether NADH or NADPH was the cofactor. These findings highlight the value of rational design, enhance our understanding of HMGR-cofactor recognition mechanisms, and provide a foundation for future efforts to optimize and engineer HMGR for broader cofactor flexibility. Full article

6-PPD quinone (6-PPDQ) is a derivative from 6-PPD, an antioxidant added in tires. Leucine is an important amino acid that needs to be obtained from the diet. In Caenorhabditis elegans, we examined the effect of 6-PPDQ exposure at environmentally relevant concentrations (ERCs) on the content of leucine and underlying mechanisms. In nematodes, 0.1–10 μg/L of 6-PPDQ decreased leucine content. The expression of the aat-1-encoding amino acid transmembrane transporter was decreased by 0.1–10 μg/L of 6-PPDQ, and leucine content was reduced by aat-1 RNAi. Meanwhile, the expression of bcat-1-encoding branched-chain amino acid transferase was increased by 0.1–10 μg/L of 6-PPDQ, and leucine content was increased by bcat-1 RNAi. Additionally, the expressions of dbt-1 and ivd-1 encoding two enzyme genes governing NADH and FADH₂ generations were decreased by 0.1–10 μg/L of 6-PPDQ, and their expressions in 6-PPDQ exposed nematodes were increased by bcat-1 RNAi. After 6-PPDQ exposure, NADH content was reduced by dbt-1 RNAi, and FADH₂ content was reduced by ivd-1 RNAi. Moreover, 6-PPDQ-induced mitochondrial dysfunction and other aspects of toxicity (such as intestinal ROS generation and lipofuscin accumulation, inhibited locomotion, and reduced brood size) were suppressed by bcat-1 RNAi and strengthened by dbt-1 and ivd-1 RNAi. The 6-PPDQ-induced toxicity and the decrease in dbt-1 and ivd-1 expressions could be inhibited by following leucine (5 mM) treatment. Our results demonstrate the important association of leucine adsorption and catabolism with 6-PPDQ toxicity induction. Full article

Ex situ machine perfusion has emerged as a pivotal technique for organ preservation and pre-transplant viability assessment, where the real-time monitoring of mitochondrial biomarkers—flavin mononucleotide (FMN) and nicotinamide adenine dinucleotide (NADH)—could significantly mitigate ischemia-reperfusion injury risks. This study develops a non-invasive optical method combining fluorescence and UV-visible spectrophotometry to quantify FMN and NADH in hypothermic oxygenated perfusion media. Calibration curves revealed linear responses for both biomarkers in absorption and fluorescence (FMN: λ_ex = 445 nm, λ_em = 530–540 nm; NADH: λ_ex = 340 nm, λ_em = 465 nm) at concentrations < 100 μg mL⁻¹. However, NADH exhibited nonlinear fluorescence above 100 μg mL⁻¹, requiring shifted excitation to 365 nm for reliable detection. Spectroscopic analysis further demonstrated how perfusion solution composition alters FMN/NADH fluorescence properties, with consistent reproducibility across media. The method’s robustness was validated through comparative studies in clinically relevant solutions, proposing a strategy for precise biomarker quantification without invasive sampling. These findings establish a foundation for real-time, optical biosensor development to enhance organ perfusion monitoring. By bridging spectroscopic principles with clinical needs, this work advances translational sensor technologies for transplant medicine, offering a template for future device integration. Full article

This study aims to clarify the dose–response relationship between dietary β-carotene levels and gonadal pigment deposition and regulation mechanisms related to the carotenoid synthesis of Strongylocentrotus intermedius based on a 60-day feeding trial and subsequent transcriptome analysis. Adult sea urchins (initial weight: 9.33 ± 0.21 g) of three cages were given one of the dry feeds with different doses of β-carotene (0 mg/kg, 150 mg/kg, 300 mg/kg) or fresh kelp (Saccharina japonica). The results indicated that the weight gain rate (WGR) of sea urchins increased with the addition of β-carotene, with that of the C300 group being markedly higher than that of the C0 group. The addition of β-carotene significantly improved the redness (a*) and yellowness (b*) values of the gonads, with sea urchins in the C300 group exhibiting closest gonad coloration to those in the kelp-fed group. Meanwhile, β-carotene and echinenone in the gonads of the C300 group showed the highest contents, reaching 1.96 μg/kg and 11.97 μg/kg, respectively. Several differential genes, enriched in the pathways of steroid biosynthesis, oxidative phosphorylation, and ubiquitination, were screened based on transcriptome analysis. Real-time PCR further demonstrated that β-carotene significantly upregulated the expression of cholesterol 25-hydroxylase (CH25H), NADH dehydrogenase subunit 1 (ND1), NADH dehydrogenase subunit 2 (ND2), and NADH dehydrogenase subunit 4 (ND4) while it downregulated the expression of 24-dehydrocholesterol reductase (DHCR24). These results showed that 300 mg/kg β-carotene significantly increased the WGR, redness, and yellowness values, as well as the contents of β-carotene and echinenone in the gonads of S. intermedius. On the one hand, dietary β-carotene increased NADH enzyme activity, which participates in echinenone synthesis by donating electrons for the transformation of β-carotene to echinenone synthesis. On the other hand, the addition of β-carotene inhibited cholesterol synthesis by increasing the expression of CH25H and decreasing the expression of DHCR24, which could in turn increase the fluidity and permeability of the cell membranes and the transport efficiency of β-carotene and echinenone from the digestive tract to the gonads. These results provided fundamental insights into the production of sea urchin gonads with market-favored colors. Full article

Obesity has been implicated in the induction of oxidative stress, which is thought to contribute to the pathogenesis of various cardiovascular diseases, including cardiac hypertrophy. However, the redox status during the early stages of cardiac hypertrophy remains inadequately characterized. In this study, we administered a high-fat diet (HFD) to C57BL/6N mice for 12 weeks. We investigated the expression of biomarkers associated with hypertrophy and oxidative stress, including lipid peroxidation, protein carbonylation, and the redox couples NADH/NAD⁺, NADPH/NADP⁺, and GSH/GSSG. Additionally, we assessed the expression levels and enzymatic activities of catalase, glutathione peroxidase, glutathione reductase, and superoxide dismutase. Following 12 weeks on a HFD, mice exhibited obesity and a 10% increase in the heart weight/tibia length ratio, together with an upregulation in the mRNA levels of β-myosin heavy chain, brain natriuretic peptide, and regulator of calcineurin 1, isoform 4. There was also a significant increase in NOX4 content in the heart of these animals; however, we observed no rise in protein carbonylation and a decrease in lipid peroxidation products. As for the redox couples, the GSH/GSSG ratio nearly doubled, while the NADH/NAD⁺ and NADPH/NADP⁺ ratios remained stable. All antioxidant enzyme mRNAs examined showed increased expression; however, only glutathione reductase showed higher activity. Our findings suggest that reductive stress is predominant within the cardiac environment of these animals. Full article

Sterlet is a perishable product; therefore, its freshness monitoring and shelf-life evaluation are important. In this study, a series of analytical techniques named physicochemical, microbiological, sensory, colorimetric, and mid-infrared and fluorescence spectroscopies were applied on Sterlet (Acipenser ruthenus) samples during 18 days of storage at 4 °C. The water content increased from 72.8 g/100 g on day 1 to 77.81 g/100 on day 14. Regarding the peroxide value (PV), the initial value was 4.17 meq/kg of Sterlet on day 1, reaching a maximum on day 4 (4.9 meq/kg of Sterlet), and then it decreased gradually, attaining a value of 0.7 meq/kg of Sterlet on day 18. Generally, the thiobarbituric acid reactive substance (TBARS), total viable count (TVC) and psychrotrophic count (PTC) increased during the storage time and increased from 0.03 to 0.13 MDA eq./kg of Sterlet sample, 2.27 to 9.09 log10 CFU/g, and 2.18 to 9.15 log10 CFU/g, respectively, on day 1 and 18, respectively. The microbiological and sensory analyses indicated that Sterlet samples were acceptable for human consumption up to 7 days of storage at 4 °C. This result was confirmed by fluorescence measurements, since the principal component analysis (PCA) applied to the NADH and MIR spectra allowed for a clear differentiation between Sterlet samples aged 7 days or less from the others. This trend was confirmed by the factorial discriminant analysis (FDA) applied to the NADH and MIR spectra, since a correct classification with leave-one cross-validation of 94.44% was observed. In addition, the heatmap of the Pearson correlation coefficients showed high correlations between overall acceptability and microbiology parameters and the structural properties of Sterlet samples during storage, indicating that the modifications observed at the macroscopic level were related to those notedat the molecular scale. Full article

Tephritidae is an economically important family among Diptera that also exhibits high diversity, biogeographical distribution, and different lifestyles. Despite the recent release of genomes and mitochondrial genome sequences of various species of the family, the evolutionary history of the group and the origin of host adaptation within it remain poorly understood. We undertook a whole-mitochondrial-genome study covering molecular variation at the mitochondrial level by analyzing 10 new mitochondrial genomes obtained from genomic data reported and downloaded from the SRA database from NCBI, analyzed in FastQC and assembled through MITGARD, and 44 mitogenomes available in the Organelle—Refseq database, in total representing 4 subfamilies, 9 tribes, 13 genera, and 54 species. We determined compositional asymmetry and codon usage patterns across the different subfamilies analyzed by using DNASp6 and CAICal. We found high evolutionary rates in the NADH genes, which could play an important role in the adaptation of species to different hosts and environmental variation. By using maximum likelihood phylogenetic reconstruction obtained by IQTREE and ModelFinder, and lifestyle and distribution data of the included species, we considered a generalist feature, explained as possible predominant adaptation in some members of the family. This study in Tephritidae tries to demonstrate possible patterns among molecular variability in mitogenomes, adaptations, and lifestyles. Our findings suggest that selection pressures on certain NADH genes may be linked to host specificity in some Tephritidae species, providing evolutionary insights into how molecular evolution drives ecological adaptation or biogeographical diversity, probably in response to changing environmental conditions and host–parasite co-evolution across taxa. Full article

Antibiotics are indispensable in medical patient care, yet they may elicit off-target effects, particularly by affecting mitochondrial function. This study investigates three commonly used antibiotics, gentamicin, ciprofloxacin, and amoxicillin, for their direct effects on mitochondrial respiration and membrane potential. Using high-resolution respirometry, we show that gentamicin and ciprofloxacin markedly increase mitochondrial leak respiration in permeabilized human embryonic kidney cells, suggesting alterations in the mitochondrial inner membrane. This is supported by a gentamicin-induced decrease in mitochondrial membrane potential. Especially gentamicin, but also ciprofloxacin, dose- and time-dependently inhibit oxidative phosphorylation and the mitochondrial electron transfer capacity, pronouncedly in the NADH-linked but also in the succinate-linked pathway. Furthermore, gentamicin decreases Complex IV (CIV) activity in a time-dependent fashion. In contrast, amoxicillin has no significant effect on mitochondrial respiration. These findings emphasize the importance of evaluating the potential direct toxicity of antibiotics on mitochondria, as they are most critical off-target sites. High-resolution respirometry provides a powerful approach to characterize such effects early in the drug development process. Full article