Search Results (80)

Lane detection is a fundamental perception task for autonomous driving and intelligent transportation systems. Although existing methods have achieved promising performance, many of them mainly focus on individual lane instances and insufficiently exploit the structural relationships among lanes, such as relative ordering, geometric continuity, and spatial parallelism. This limitation may lead to broken lanes, ordering errors, and geometric inconsistencies in complex road scenarios. To address these issues, we propose TPDNet, a topology-aware lane detection framework that incorporates structural reasoning into the detection pipeline at three complementary levels. First, a Topology-aware Perception Reasoner (TPR) is introduced at the feature level to capture relational dependencies among lane features and enhance the representation of global road topology. Second, a Topology-Decoupled Head (TDH) is designed at the prediction level to decouple geometric regression from lane classification, thereby reducing task interference and improving prediction stability. Third, a Topology Consistency Loss (TCL) is formulated as a complementary supervision term to encourage smoothness and ordering consistency in predicted lanes. Extensive experiments on three public benchmarks demonstrate the effectiveness of the proposed method. On CULane, TPDNet achieves an F1@50 of 81.46 with ResNet101 and remains competitive with the strongest compared methods, while showing improved robustness in challenging scenarios such as Curve and Dazzle light. On TuSimple, TPDNet obtains an F1 score of 98.01 among the compared methods, while maintaining competitive accuracy. On CurveLanes, TPDNet achieves an mF1 of 58.74, exceeding the strongest baseline by 4.98 points. These results suggest that topology-aware reasoning can improve the generalization capability of lane detection and help produce more structurally coherent lane predictions under diverse road conditions. Full article

Cancer cells, like yeast, use fermentation despite the presence of oxygen, a phenomenon called aerobic glycolysis. The advantage is that it maintains many C-C bonds of glucose, allowing highly proliferating cells to produce the biomolecules that are necessary for cytokinesis. However, aerobic glycolysis is less energy-efficient than respiration, and it must operate at high frequency and produces large amounts of lactate, which modifies and stimulates DNA repair enzymes via lysine lactylation. This makes cancer cells resistant to radiotherapy, which requires a combination with chemotherapy using drugs that inhibit DNA repair. However, this converts healthy cells to cancer cells, indicating that research is still required regarding the relationship between glycolysis and cancer. Using yeast as a model, we discovered that the glycolytic enzymes TPI and GAPDH (Tpi1p and Tdh1-3p in yeast) interact with the DNA damage-dependent Checkpoint Rad9p (53BP1/BRCA1/MDC1 in humans). We propose that Tpi1p and Tdh1-3p override Rad9p, allowing cells with damaged DNA to proliferate. We isolated tpi and gapdh mutant strains that are deficient in DNA repair. While the tpi mutant strain has lower enzymatic activity, the gapdh mutant strains have normal enzymatic activity, confirming previous reports that GAPDH moonlights in the DNA damage response. Full article

V. parahaemolyticus has several virulence factors, including thermostable direct hemolysin (TDH), TDH-related hemolysin (TRH), and two separate type III secretion systems (T3SSs), T3SS1 and T3SS2. T3SS1 is responsible for cytotoxicity, primarily through the activity of its effector VP1680. To gain a detailed understanding of the relationship between the amount of effector, its expression timing, and cytotoxicity, a system is required to regulate protein expression levels and timing. In the present study, we developed an effector protein expression system controlled by an arabinose-dependent transcription factor and found that cytotoxicity toward mammalian cells increased in a VP1680-dependent manner. To ensure specific protein degradation, we also established a targeted protein degradation system, including VP0917 (ClpP) and VP0918 (ClpX)-, or VP0917 and VP1014 (ClpA)-mediated degradation of ssrA-tagged proteins (proteins bearing the C-terminal degradation tag encoded by tmRNA). By combining these systems, more than 50% of the targeted protein could be degraded within 20 min. As a byproduct of creating the systems, we obtained an enhanced green fluorescent protein variant that emits strong fluorescence in V. parahaemolyticus. The protein degradation system developed in this study has demonstrated the potential to control intracellular protein levels to a certain extent. Moreover, experimentally controlling intracellular protein levels will allow for a more detailed examination of the relationship between protein quantity and cellular phenotype, potentially overcoming the limitations of the “all-or-nothing” model. Full article

Background: The development of lung cancer is strongly influenced by oxidative stress (OS), which results when the balance between oxidants and antioxidants is disturbed. Evaluation of both specific redox markers such as thiol/disulfide homeostasis (TDH) and overall indicators including total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) may provide a more comprehensive view of oxidative imbalance in lung cancer. We examined OS indices and TDH in patients with lung cancer versus healthy controls. Methods: Eighty participants were enrolled, consisting of 40 patients with newly diagnosed lung cancer and 40 age- and sex-matched healthy controls. Serum levels of native thiol (NT), total thiol (TT), and disulfide were determined using an automated spectrophotometric method. Additionally, TAS, TOS, and the OSI were evaluated to provide an overall assessment of oxidative balance. Routine hematological and biochemical parameters were compared between groups. Results: White blood cell and neutrophil counts were notably higher in lung cancer patients compared with controls (p < 0.05). NT and TT levels were remarkably decreased, whereas disulfide levels, TOS, and OSI were significantly elevated in the lung cancer group (p < 0.05). TAS levels tended to be lower in patients, although not reaching statistical significance. No significant association was observed between oxidative parameters and tumor stage or localization. Conclusions: Patients with lung cancer exhibited a marked oxidative imbalance, characterized by elevated oxidant burden and impaired TDH. Combined assessment of TAS, TOS, OSI, and thiol/disulfide parameters may provide valuable insight into the oxidative pathophysiology of lung cancer and hold potential as complementary biomarkers for disease evaluation. Further large scale studies are needed to confirm these findings. Full article

Background: Allergic rhinitis (AR) is a chronic immunoglobulin E (IgE)-mediated inflammatory disorder triggered by aeroallergens. Oxidative stress (OS) is increasingly recognized as a key factor in AR pathophysiology. This study aimed to investigate dynamic thiol–disulfide homeostasis (TDH) and OS markers in AR patients compared to healthy controls. Methods: Sixty-two participants (31 AR patients, 31 controls) were enrolled. Hematological and biochemical parameters were measured. OS markers including total thiol (TT), native thiol (NT), disulfide, total antioxidant status (TAS), total oxidant status (TOS), and oxidative stress index (OSI) were assessed. Correlations between OS markers and laboratory parameters were analyzed. Receiver operating characteristic (ROC) analysis evaluated the diagnostic performance of OS markers. Results: TT and NT levels were significantly lower in AR patients, whereas disulfide, disulfide/NT and disulfide/TT ratios, TOS and OSI were significantly higher. TAS levels were slightly lower in AR patients. TT and NT correlated positively with eosinophil counts and negatively with monocyte, platelet, AST, and creatinine levels. ROC analysis indicated strong diagnostic potential: TT (AUC = 0.749, cutoff 415 µmol/L, sensitivity 90%, specificity 61%), NT (AUC = 0.786, cutoff 373.2 µmol/L, sensitivity 90%, specificity 71%), and disulfide (AUC = 0.690, cutoff 20 µmol/L, sensitivity 74%, specificity 61%). Conclusions: AR patients exhibit disrupted TDH and elevated OS. These markers may serve as sensitive indicators of oxidative imbalance, offering potential diagnostic and therapeutic insights into AR management. Full article

Antibiotic resistance is an increasing concern in modern healthcare. Therefore, it is important to identify novel antimicrobial agents and new molecular targets for such compounds. Here, we describe the identification of an N-pyridylpyrazolone derivative, 4-(2-aminoethyl)-2-(pyridin-2-yl)-1,2-dihydro-3H-pyrazol-3-one dihydrochloride (compound 1), which is effective against Gram-positive and Gram-negative bacteria and inhibits the enzymatic activity of Escherichia coli L-threonine dehydrogenase (TDH). To characterize its interaction with compound 1, TDH was overexpressed in E. coli. The recombinant enzyme was shown to exist in dilute solution in equilibrium between dimeric and tetrameric forms, with a K_d value for the dimer/tetramer transition of 3 ± 1 nM, and to bind L-threonine cooperatively with a Hill coefficient of 1.4. Compound 1 acted as a partial mixed inhibitor of TDH with an EC₅₀ value of 47 ± 16 µM and did not affect the equilibrium between oligomeric states. Altogether, these findings identify compound 1 as a promising starting point for the development of novel antibiotics and as a tool compound for studying the functional properties of TDH. Full article

Enteritis is a common and recurrent disease in shrimp aquaculture, causing significant economic losses and management challenges. However, its specific causative pathogen remains unclear. Here, a pathogen strain, Vibrio parahaemolyticus VSP1, was directly isolated from shrimp with enteritis, and its pathogenicity and genomic characteristics were analyzed. Diseased shrimp exhibited lethargy, empty gut, hepatopancreatic atrophy, and severe intestinal damage. The gut bacterial community of diseased shrimp differed significantly from healthy shrimp (PERMANOVA, p < 0.05), with a 129% increase in Vibrio relative abundance. Nine Vibrio operational taxonomic units (OTUs) were enriched in diseased shrimp, and the dominant OTU1 shared 100% 16S rRNA identity with VSP1. VSP1 grew rapidly, utilized diverse carbon sources, and induced enteritis symptoms in over 90% of challenged shrimp. Genome analysis revealed 98.34% average nucleotide identity with V. parahaemolyticus ATCC 17802 and identified 156 putative virulence-related genes, mainly related to adherence, motility, and secretion systems. Unlike the strain ATCC 17802, VSP1 lacks thermostable direct hemolysin (TDH) and type III secretion system 2 (T3SS2), but contains alternative virulence factors such as Yersinia-like type IV pili and lipooligosaccharides, suggesting a distinct virulence strategy. This study identifies the pathogen responsible for shrimp enteritis and provides a foundation for targeted control strategies in aquaculture. Full article

CaMK4, a calcium/calmodulin-dependent protein kinase, is an important mediator of cellular signal transduction, yet its role in the regulation of skeletal muscle satellite cells (MuSCs) in goats has remained unclear. In this study, CaMK4 overexpression and knockdown models were established, and integrated transcriptomic and proteomic analyses were performed to systematically elucidate its regulatory network. CaMK4 overexpression altered key pathways associated with cell proliferation and muscle development, including cAMP, PI3K-Akt, and actin cytoskeleton regulation, while proteomic data highlighted calcium signaling and JAK-STAT pathways. Conversely, CaMK4 knockdown enhanced MuSC proliferation by upregulating cell cycle-related genes and proteins. Integrated analyses further identified that Galectin-9 (LGALS9), Collagen triple helix repeat containing-1 (CTHRC1), Hyaluronan Synthase 1 (HAS1), and L-Threonine Dehydrogenase (TDH) may serve as potential key nodes regulating cell cycle, apoptosis, and metabolic control. This suggests a regulatory role for CaMK4. Collectively, these findings provide a mechanistic framework for understanding CaMK4 function in ruminant muscle development and may offer insights for improving goat muscle growth, meat quality traits, and production efficiency. Full article

Background and Objectives: Sarcopenic obesity (SO), characterized by the coexistence of excess adiposity and reduced muscle mass/function, is associated with adverse outcomes in older adults. Oxidative stress has been implicated in the pathogenesis of both obesity and sarcopenia. This study aimed to evaluate the association between thiol/disulfide homeostasis (TDH), ischemia-modified albumin (IMA), and SO in obese older adults. Materials and Methods: In this cross-sectional study, 132 obese individuals aged ≥65 years were enrolled from a geriatrics outpatient clinic. SO was defined based on the ESPEN/EASO criteria, incorporating anthropometric, body composition, and muscle function measures. Serum native and total thiol levels, disulfide concentrations, and IMA were assessed. Logistic regression identified independent predictors of SO, and ROC analysis evaluated the discriminatory power of oxidative parameters. Results: SO was present in 15.2% (n = 20) of participants. Patients with SO exhibited significantly lower native (p = 0.003) and total thiol levels (p < 0.001), and higher disulfide/native thiol (p = 0.009) and disulfide/total thiol ratios (p = 0.009). IMA levels were slightly elevated in SO but not significantly different (p = 0.13). In multivariable regression, age and disulfide/native thiol ratio were independent predictors of SO (OR = 5.71, p = 0.041). ROC analysis showed that disulfide/native thiol ratio had moderate predictive accuracy (AUC = 0.684, p = 0.008), with a cut-off > 6.63 yielding 92.86% specificity. Conclusions: Older adults with SO exhibit disrupted redox balance, as evidenced by altered TDH parameters. The disulfide/native thiol ratio may serve as a useful oxidative biomarker for identifying SO. These findings highlight the potential role of oxidative stress in SO and warrant further research into targeted antioxidant strategies. Full article

Vibrio parahaemolyticus is a major foodborne pathogen worldwide, responsible for seafood-associated poisoning. Among its toxin genes, tdh2 is the most critical. To investigate the role of tdh2 in V. parahaemolyticus under gastrointestinal conditions, we constructed tdh2 deletion and complementation strains and compared their survival under acid (pH 3 and 4) and bile stress (2%). The results showed that tdh2 expression was significantly upregulated under cold (4 °C) and bile stress (0.9%). Survival assays and PI staining revealed that the tdh2 mutant strain (VP: △tdh2) was more sensitive to acid and bile stress than the wild-type (WT), and this sensitivity was rescued by tdh2 complementation. These findings suggest that tdh2 plays a protective role in enhancing V. parahaemolyticus tolerance to acid and bile stress. In the VP: △tdh2 strain, seven genes were significantly upregulated and six were downregulated as a result of tdh2 deletion. These genes included VPA1332 (vtrA), VPA1348 (vtrB), VP2467 (ompU), VP0301 and VP1995 (ABC transporters), VP0527 (nhaR), and VP2553 (rpoS), among others. Additionally, LC-MS/MS analysis identified 12 differential metabolites between the WT and VP: △tdh2 strains, including phosphatidylserine (PS) (17:2 (9Z,12Z) /0:0 and 20:1 (11Z) /0:0), phosphatidylglycerol (PG) (17:0/0:0), flavin mononucleotide (FMN), and various nucleotides. The protective mechanism of tdh2 may involve preserving cell membrane permeability through regulation of ompU and ABC transporters and enhancing electron transfer efficiency via regulation of nhaR. The resulting reduction in ATP, DNA, and RNA synthesis—along with changes in membrane permeability and electron transfer due to decreased FMN—likely contributed to the reduced survival of the VP: △tdh2 strain. Meanwhile, the cells actively synthesized phospholipids to repair membrane damage, leading to increased levels of PS and PG. This study provides important insights into strategies for preventing and controlling food poisoning caused by tdh⁺ V. parahaemolyticus. Full article

Vibrio parahaemolyticus is the leading bacterial cause of gastroenteritis associated with aquatic food consumption globally. This study aimed to determine the prevalence of V. parahaemolyticus in aquatic foods from Huzhou and to identify the serotypes, antimicrobial resistance, virulence factors, and genetic relatedness of the strains. A total of 306 isolates were detected from 1314 aquatic food samples from 2022 to 2024. The results indicated that the most prevalent serotypes were O1:KUT (17.0%), O2:K28 (13.7%), and O2:KUT (13.1%). Multilocus sequence typing analysis divided the 306 isolates into 175 sequence types (STs), and the predominant sequence type was ST864 (3.3%). Antimicrobial susceptibility tests showed that 2.6% of isolates were multidrug resistant. High resistance was observed to ampicillin (64.7%) and streptomycin (44.4%). A total of seven antimicrobial categories of resistance genes were identified, and the resistance gene bla_CARB was detected in all isolates. The virulence genes tdh and trh were found in 16 (5.2%) and 12 (3.9%) isolates, respectively. In addition, we observed that all the 306 V. parahaemolyticus isolates encode type III secretion systems 1. The phylogenomic analysis based on the whole-genome sequence revealed that the 306 isolates were divided into four clusters. Our findings broaden perspectives on V. parahaemolyticus genetic diversity and enhance our ability to assess the potential risks of its spread. Full article

2-Phenylethanol (2-PE), an aromatic alcohol with a rose-like fragrance, is widely used in the food, pharmaceutical, and high-end cosmetic industries. In this study, a high-yield 2-PE-producing strain was isolated and identified as Saccharomyces cerevisiae based on morphological characterization and taxonomic identification. Fermentation medium components (carbon and nitrogen sources) were optimized through single-factor experiments in shaking flasks, and fermentation medium with 40 g/L glucose, 5 g/L malt extract, 1.75 g/L corn steep liquor, 2.5 g/L yeast extract, 5 g/L malt extract, 1.75 g/L corn steep liquor was considered suitable for 2-PE production. RT-qPCR results indicated that corn steep liquor activates expression of genes related to the shikimate pathway and Ehrlich pathway (pha2, aro4, aro8, and aro9), thereby promoting the synthesis of 2-PE through these pathways. Excess yeast extract inhibited the expression of aro8 and aro9, while enhancing the expression of tdh3 and adh2, thus promoting the de novo synthesis of 2-PE. Furthermore, fermentation in a 5 L bioreactor was applied to investigate the effects of feeding strategies, inoculum proportion, and pH on 2-PE production. With a pH of 5.5 and10% inoculum proportion, the supplementation of the substrate L-Phe led to a 2-PE production of 4.81 g/L after 24 h of fermentation. Finally, in situ product recovery (ISPR) techniques was applied to alleviate 2-PE cytotoxicity, achieving a production of 6.41 g/L. This process offers a promising strategy for producing 2-PE efficiently and naturally, paving the way for further industrial applications in food, pharmaceutical, and cosmetic sectors. Full article

Vibrio alginolyticus, a common Gram-negative opportunistic pathogen of marine animals and humans, is known for its rapid growth in organic-matter-rich environments. However, it remains unclear how it incorporates metabolic pathways in response to diverse carbon and nitrogen sources and rapidly alters gene expression. Increasing evidence suggests that post-transcriptional regulation by RNA-binding proteins and small RNAs (sRNAs) plays a crucial role in bacterial adaptation and metabolism. CsrA (carbon storage regulator A), a conserved post-transcriptional regulator in Gammaproteobacteria, is poorly characterized in Vibrio species. Using integrated transcriptomic and proteomic analyses, we found that CsrA alters the expression of 661 transcripts and 765 protein transcripts in V. alginolyticus, influencing key pathways including central carbon metabolism, amino acid metabolism and transport, quorum sensing, and bacterial secretion systems. Through directed CsrA-RNA EMSAs, we identified several direct mRNA targets of CsrA, including gltB, gcvP, aceE, and tdh, as well as secretion system components (tagH, tssL, yopD, and sctC). Notably, CsrA also directly regulates rraA, a key modulator of ribonuclease activity, suggesting a broader role in RNA metabolism. Our findings establish CsrA as a global regulator in V. alginolyticus, expanding the known targets of CsrA and providing new insights into its regulatory roles. Full article

2,3,5-Trimethylpyrazine (TMP) is an alkyl pyrazine with broad application prospects in the fields of food additives and medicine. L-threonine-3-dehydrogenase (TDH) is a key enzyme in the biosynthesis pathway of TMP. To explore the efficient and environmentally friendly production method of TMP, we constructed recombinant strains overexpressing the BlTDH gene and its mutant BlTDH (N157A) using Bacillus licheniformis YC7. The TMP yield of recombinant strains with pHT01-BlTDH (N157A) reached 15.35 ± 1.51 mg/L, which was significantly higher than that of strains with pHT01-BlTDH (9.86 ± 1.24 mg/L) and strains with vector pHT01 (2.35 ± 0.58 mg/L). To further increase the TMP yield of strain YC7/pHT01-BlTDH (N157A), the fermentation process was optimized by single-factor experiments, and the response surface test was conducted using the Box–Behnken design. The results revealed that the substrate ratio, IPTG concentration and fermentation time had significant effects on the yield of TMP, and the degree of influence was substrate ratio > fermentation time > IPTG concentration. The optimization results of response surface indicated that the optimal fermentation conditions were as follows: substrate ratio of 1:2, IPTG concentration of 1.0 mM, and fermentation time of 4 d. Under these conditions, the TMP yield reached 44.52 ± 0.21 mg/L, which was around 0.005 mg/L different from the predicted value (45.515 mg/L), and increased by 29.17 mg/L compared with the initial condition. The optimization of fermentation conditions significantly increased the yield of TMP produced by recombinant strains, which provided a theoretical basis and strain resources for industrial production of TMP. Full article

Background/Objectives: Vibrio parahaemolyticus is a marine bacterium and a major cause of food poisoning worldwide, primarily associated with gastric illnesses such as gastroenteritis. This study aimed to investigate the distribution, antibiotic resistance, and virulence genes of V. parahaemolyticus present in shellfish and seawater of the southern coast of Korea, a major shellfish harvesting area. Methods: Shellfish and seawater samples were collected monthly in 2023 from 24 coastal sites in Korea. V. parahaemolyticus was isolated and identified using the MPN method, biochemical tests, MALDI-TOF mass spectrometry, and 16S rRNA sequencing. Antimicrobial susceptibility was tested for 673 isolates using the Sensititre MIC system, and virulence genes (tdh and trh) were detected by PCR. Results:V. parahaemolyticus had a detection rate of 18.2–58.3% in shellfish and 8.3–50% in seawater samples. Among the isolates, 97.9% and 97.3% were resistant to ampicillin and colistin, respectively, while 8.3% showed resistance to four or more antibiotics. The virulence genes tdh and trh were detected in 0.45% and 3.34% of shellfish samples and 1.23% and 4.46% of seawater samples, respectively. Conclusions: These findings will help implement appropriate precautionary measures to prevent potential human health risks arising from exposure to multidrug-resistant or pathogenic V. parahaemolyticus. Full article