Search Results (19,538)

Poly(lactic acid) (PLA) films containing two different hemp-derived essential oils (EOs), Carmagnola CS (Carm) and Futura 75 (Fut), at 1, 5, and 10% wt were successfully produced via solvent casting for packaging applications. The influence of EO presence, type, and concentration on the chemical, morphological, and thermal properties of the PLA-based films was investigated. In addition, radical-scavenging activity, water transport properties, and antimicrobial performance were evaluated to assess the effect of EOs on the structural and functional characteristics of the resulting packaging materials. FTIR spectroscopy confirmed the successful incorporation of the hemp essential oils Carm and Fut into the polymer matrix, with a concentration-dependent effect that is more pronounced for Fut than for Carm. In the second heating run, evaluated by DSC measurements, both EOs lowered T_g from 60.3 °C (PLA) to 52.0 °C for PLA_10 Carm and 55.1 °C for PLA_10 Fut. The EOs act as plasticizers in the PLA matrix, improving the deformation at break. Gas barrier measurements showed that permeability decreased from 3027 ± 300 Barrer (PLA) to (2499 ± 44) Barrer in PLA_10 Carm and 2623 ± 130 Barrer in PLA_10 Fut, with a corresponding reduction in diffusivity. The barrier improvement factor reached 17% for Carm and 15% for Fut, confirming the enhanced barrier performance of PLA_EOs films. DPPH assays showed that PLA_EOs films retained most of the antioxidant activity of the free oils, with only a 10–15% reduction for PLA_Fut and no significant loss for PLA_Carm after one week. After one month, the activity of Carm in PLA film decreased by 18%, whereas the performance of its free form remained unchanged, confirming the superior and more stable radical scavenging capacity of Carm compared to Fut. Overall, the study demonstrates that hemp essential oils can be effectively integrated into PLA without compromising structural integrity, while preserving antioxidant performance and enhancing water barrier properties, supporting their potential as sustainable active packaging components. Full article

This study systematically evaluated the taxonomic relationship between Streptococcus taoyuanensis ST2^T and Streptococcus caecimuris CLAAV18^T. Comparative genomic analysis revealed a high 16S rRNA gene sequence similarity of 99.6%, with the two strains clustering closely in the 16S rRNA-based phylogenetic tree. The genetic relatedness was further validated by Multi-Locus Sequence Typing (MLST) analysis: assessments of seven conserved housekeeping genes (atpD, gapA, gyrB, GdhA, recA, dnaK, and sdhA) demonstrated complete concordance in target fragment lengths (ranging from 33 bp to 121 bp). No size polymorphisms, insertions, or deletions were detected, indicating a highly conserved core genome. At the whole-genome level, the Average Amino Acid Identity (AAI), Average Nucleotide Identity (ANI), and digital DNA-DNA hybridization (dDDH) values between the two strains were 96.8%, 95.7%, and 84.6%, respectively. These values significantly exceed the established thresholds for species delineation (AAI: 95.5%; ANI: 95%; dDDH: 70%), providing robust genomic evidence that both strains belong to the same species. Furthermore, phenotypic testing confirmed nearly identical physiological characteristics, with only minor biochemical variations. Based on the integration of phylogenetic, genomic, and phenotypic evidence, we formally propose Streptococcus taoyuanensis as a later heterotypic synonym of Streptococcus caecimuris. Full article

Background/Objectives: Antimicrobial agents have revolutionized disease management in humans and animals; however, their misuse and overuse have accelerated the emergence and spread of antimicrobial resistance (AMR) and antimicrobial resistance genes (ARGs). Dairy farms are recognized as potential hotspots for ARG dissemination, particularly through Escherichia coli, which acts as a reservoir and vector of ARGs, enabling their horizontal transfer via plasmids and other mobile genetic elements. This study aimed to characterize the genomic diversity, ARG profiles, plasmid content, and phylogenetic relationships of E. coli isolated from dairy farm environments and milk using whole-genome sequencing. Methods: A total of 31 E. coli isolates recovered from soil, effluent, cow dung, and milk samples underwent deoxyribonucleic acid extraction, library preparation, and sequencing on the Illumina MiSeq platform, followed by comprehensive bioinformatic analysis. Results: The E. coli isolates exhibited 20 distinct sequence types, including one novel sequence type. Plasmids were detected in 71% of the isolates, with the IncF plasmid family being the most predominant. Furthermore, 12 ARG groups were identified, with β-lactam resistance genes detected in 67.7% of isolates. Notably, blaCTX-M genes were identified in all phenotypically confirmed extended-spectrum β-lactamase-producing isolates. Additional ARGs, including those conferring resistance to tetracyclines (tet(A), tetX4), quinolones (qnrS1), aminoglycosides (aph, aad, ant), and folate pathway inhibitors (dfr and sul), were widely distributed throughout the samples. Phylogenetic analysis revealed clustering of isolates from different sample types, particularly among ST58 isolates, suggesting cross-environmental transmission. Conclusions: This study demonstrates that E. coli from dairy farm environments harbor diverse ARGs and plasmids, confirming their role as reservoirs of AMR. These findings underscore the importance of prudent antimicrobial use, routine genomic surveillance, and enhanced biosecurity measures to limit cross-environmental transmission. Full article

Background/Objectives: Enteropathogenic Escherichia coli O157:H7 infection disrupts intestinal homeostasis, causing dysbiosis, barrier dysfunction, and inflammation. This study aimed to evaluate the protective efficacy and mechanisms of a novel probiotic, Bacteroides thetaiotaomicron type strain ATCC 29148, isolated from goat feces, against E. coli O157:H7-induced colitis. Methods: This study assessed the protective potential of the probiotic strain Bacteroides thetaiotaomicronBT6 and BT7 in vitro for GI tolerance, adhesion, and no adverse effects were observed. For the in vivo experiment, male C57BL/6J mice were divided into groups treated with Bacteroides thetaiotaomicron (BT6), PBS, E. coli O157:H7, or a combination. We employed integrated analyses including 16S rRNA gene sequencing, antioxidant status, cytokine profiling, and short-chain fatty acid (SCFA) measurement. Results: In vitro, Bacteroides thetaiotaomicron (BT6 and BT7) showed high gastrointestinal tolerance (71.89–93.22% survival). In vivo, it significantly mitigated infection-associated weight loss and disease activity (p < 0.05). Probiotic treatment enhanced barrier integrity, reduced colonic inflammation, and modulated systemic immune responses, notably increasing anti-inflammatory IL-10 while decreasing pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 (p < 0.05). It also alleviated oxidative stress by reducing malondialdehyde (MDA) and elevating antioxidant enzymes (SOD, CAT, GSH) and ATP. Fecal SCFA profiling revealed increased propionic and butyric acid. 16S sequencing indicated that B. thetaiotaomicron (BT6) administration increased beneficial families (Lactobacillaceae, Muribaculaceae) and suppressed pathobionts. Conclusions: B. thetaiotaomicron (BT6) probiotic with potential for mitigating enteropathogenic infection, an effect mainly determined by its capacity to reestablish the intestinal epithelial barrier and enhance global host health, and modulating the inflammatory response Full article

Radish (Raphanus sativus L.) is an important vegetable resource in the food industry, generating substantial amounts of by-products during cultivation and distribution. Despite their richness in functional components, these by-products are largely underutilized. Accordingly, there is increasing interest in their valorization as functional food ingredients. This study evaluated the functional potential of radish by-products removed prior to distribution by applying cellulase pretreatment and Lactiplantibacillus plantarum fermentation individually or in combination. Radish samples were separated into leaf blade, stem, and taproot tissues and processed as untreated control, enzyme-treated, fermented, and enzyme-treated and fermented (EF) groups. The EF treatment significantly increased reducing sugar content, total polyphenols, and total flavonoids across all tissues, with the most pronounced enhancement observed in leaf by-products. In antioxidant assays, EF samples showed decreased IC₅₀ values in DPPH and ABTS radical scavenging assays and increased FRAP values, indicating superior antioxidant capacity. In lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, EF-treated leaf extracts effectively suppressed nitric oxide and intracellular ROS production without cytotoxicity and exhibited the highest GSH/GSSG ratio, suggesting improved cellular redox balance. In contrast, interleukin-6 (IL-6) secretion varied depending on tissue type and processing condition, indicating that antioxidant enhancement does not necessarily correspond to uniform cytokine regulation. Collectively, these findings demonstrate that enzymatic pretreatment combined with lactic acid fermentation serves as an effective strategy to enhance the antioxidant and cell-protective properties of radish by-products, supporting their potential use as value-added functional food ingredients. Full article

Johne’s disease, caused by Mycobacterium avium subsp. paratuberculosis (MAP), is endemic in Saudi Arabia and contributes to substantial production losses in small-ruminant herds. This study investigated MAP infection in 388 locally raised small ruminants (151 sheep and 237 goats) using IS900 real-time PCR (qPCR) on fecal samples and indirect ELISA on serum samples. Ziehl–Neelsen (ZN) staining and pathological assessment were applied as supportive tools in necropsied cases. Overall, qPCR detected MAP DNA in 135/388 animals (34.8%), with higher positivity in goats (100/237; 42.1%) than in sheep (35/151; 23.1%). ELISA detected MAP antibodies in 120/388 animals (30.9%), including 90/237 goats (37.9%) and 30/151 sheep (19.8%). Based on clinical examination (presence/absence of clinical signs), qPCR-positive animals were categorized as subclinical (n = 15; 10 goats and 5 sheep) or clinically progressed (n = 120; 90 goats and 30 sheep). Gross and histopathological findings were assessed in a necropsied subset (n = 20; 10 subclinical and 10 clinically progressed), revealing mild focal granulomatous enteritis with scant acid-fast bacilli in subclinical cases and diffuse lepromatous-type granulomatous lesions with abundant bacilli in clinically progressed animals. Genotyping and subtyping were performed on tissue-derived DNA from six necropsied cases using DMC, IS900, and F57 targets, and IS900 sequencing confirmed the circulation of both Type C/II and Type S/I MAP strains. Collectively, these findings demonstrate widespread MAP infection among small ruminants in Saudi Arabia, with higher detection rates and more pronounced pathology in goats, and highlight the genetic heterogeneity of circulating MAP strains. Full article

This study aimed to systematically elucidate the antihyperlipidemic mechanism of paeoniflorin, and we adopted an integrated multi-omics strategy to screen the key molecular targets and regulatory pathways involved in its action, followed by experimental validation to verify the potential regulatory effects of paeoniflorin on the screened targets and metabolic processes. Rats with high-fat diet-induced hyperlipidemia received paeoniflorin treatment. Liver histopathology was evaluated using hematoxylin–eosin and Oil Red O staining. Serum levels of total cholesterol, triglycerides, low-density lipoprotein cholesterol, high-density lipoprotein cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, total bile acids, activated partial thromboplastin time, prothrombin time, thrombin time, and fibrinogen were measured using a biochemical analyzer. Integrated multi-omics analyses were performed to investigate paeoniflorin’s lipid-lowering mechanism. Critical pathways and targets identified were validated using Western blotting. Paeoniflorin alleviated pathological liver damage in hyperlipidemic rats and improved blood lipid levels, coagulation function, and liver function markers. Multi-omics analyses verified that paeoniflorin downregulated the expression of TREM-1, TLR4, NF-κB, TNF-α, and IL-1β, thereby alleviating hepatic inflammation. Paeoniflorin also upregulated the expression of low-density lipoprotein receptors (LDLR), liver X receptor alpha (LXRα), and ATP-binding cassette subfamily G member 1 (ABCG1), while downregulating proprotein convertase subtilisin/kexin type 9 (PCSK9) expression, contributing to balanced cholesterol metabolism. Paeoniflorin normalized glycerophospholipid and branched-chain amino acid metabolism, which correlated with reduced inflammation and improved cholesterol metabolism. Paeoniflorin ameliorates hyperlipidemia through multitarget mechanisms, potentially by suppressing the TREM-1-TLR4-NF-κB signaling pathway to reduce inflammation and by regulating cholesterol metabolism via the PCSK9-LDLR and LXRα-ABCG1 pathways. Full article

The modulation of plant responses to abscisic acid (ABA) and/or abiotic stresses can be manipulated by the expression of ABA-responsive genes, which is affected by phytohormone ABA. While some ABA-responsive genes have been shown to regulate plant responses to ABA and/or abiotic stresses, the functions of numerous ABA-responsive genes remain unknown. Therefore, characterizing these unstudied genes would provide a practical way to identify novel regulators of plant adaptations to ABA and/or abiotic stresses. Here, we characterized four closely related unstudied ABA-responsive genes in Arabidopsis thaliana, named Arabidopsis thaliana ABA-up regulated genes (AtAUGs). We found that ABA treatment induces AtAUGs expression level, and our results in transfected protoplasts show that AtAUGs exhibit nucleus localization and downregulate the co-transfected reporter expression level. The results of ABA sensitivity assays, including seed germination, cotyledon greening, and root extension assay show that transgenic plants overexpressing AtAUGs had increased sensitivity, but ataugs mutants generated by isolating T-DNA insertion lines or through CRISPR/Cas9 gene-editing of AtAUGs had decreased sensitivity; in addition, the greatest decrease in ABA sensitivity was observed in the ataug1 ataug2 ataug3 ataug4 (ataug1234) quadruple mutants. The qRT-PCR results show that the expression levels of several Type 2C Protein Phosphatase (PP2C) genes, the key negative regulator genes of ABA signaling including PP2CA, Hypersensitive to ABA 1 (HAB1), HAB2, Highly ABA-Induced PP2C protein 3 (HAI3), ABA-Hypersensitive Germination 1 (AHG1), and ABA Insensitive 2 (ABI2) decreased in 35S:AtAUGs transgenic plants, but increased in the ataug1234 quadruple mutants. Taken together, these results suggest that AtAUGs are ABA-responsive genes, and AtAUGs positively regulate ABA responses in a redundant manner, by downregulating the expression of crucial negative regulator genes in ABA signaling. Full article

Background: Cymbidium goeringii, one of China’s traditional and valuable orchids, possesses significant ornamental and economic value. However, it is relatively sensitive to low temperature and other abiotic stresses, which severely restrict its application in landscaping and industrial development. WRKY transcription factors play important roles in plant responses to abiotic stresses, yet related research in C. goeringii remains limited. Methods: In this study, based on transcriptome data of C. goeringii under four different stresses, we identified and cloned the WRKY transcription factor gene CgWRKY53. Through bioinformatics analysis, quantitative real-time PCR, and heterologous transformation in Arabidopsis thaliana, we systematically investigated its structural characteristics, expression patterns, and function under cold stress. Results: The full-length CDS of CgWRKY53 is 1080 bp, encoding a protein of 359 amino acids with a molecular weight of 39.95 kDa. Group III subfamily of the WRKY family, possessing the conserved WRKYGQK domain and a C2HC-type zinc finger motif. CgWRKY53 is expressed in roots, pseudobulbs, leaves, and flowers of C. goeringii, with the highest expression observed in flowers. Under cold, heat, waterlogging, and ABA treatments, CgWRKY53 displayed significant changes in expression, with the most pronounced response occurring under cold stress, where its expression was significantly upregulated. Homozygous transgenic A. thaliana lines overexpressing CgWRKY53 exhibited dwarfed stature, with smaller and deformed leaves and notably shorter roots compared to wild-type plants. The overexpression lines also showed cold-sensitive phenotypes under low-temperature stress, and the expression of several cold-responsive genes was suppressed, suggesting that CgWRKY53 may act as a negative regulator in the response to cold stress. Conclusions: These results identify CgWRKY53 as a negative regulator of cold stress response in C. goeringii. This study provides important genetic resources and theoretical foundations for molecular breeding of stress-resistant orchids. Full article

The self-assembly of a novel synthesized chiral dipeptide, Boc-p-nitro-L-phenylalanyl-tyrosine, into supramolecular structures is investigated by optical absorption and photoluminescence spectroscopy as well as single crystal X-ray diffraction. The compound is a diphenylalanine derivative belonging to a family of aromatic dipeptides that spontaneously self-organize into nanostructures through molecular recognition. The dipeptide exhibits several step-like peaks in its absorption band, indicative of self-assembly into quantum-confined nanostructures. In contrast, the parent Boc-p-nitro-L-phenylalanine amino acid lacks these features, indicating that the tyrosine residue favors quantum-confined self-assembly. Crystal structure determination reveals distinct packing styles: Boc-p-nitro-L-phenylalanine forms two-dimensional hydrogen-bonded layers, while the related p-nitro-free Boc-L-phenylalanyl-tyrosine dipeptide organizes into a 3D helical columnar architecture, driven by the additional hydrogen-bonding capacity of the peptide bond and tyrosine hydroxyl group, which favors the formation of a channel-type tetragonal architecture network over the planar sheets of the monomer. Furthermore, the introduction of a tyrosine residue into the Boc-p-nitro-L-phenylalanine molecule alters its supramolecular assembly, as the dipeptide Boc-p-nitro-L-phenylalanyl-tyrosine crystallizes as a monohydrate. The water molecule present in the structure acts as a bridge, participating in a hydrogen-bonding network between the tyrosine hydroxyl groups of neighboring columns through intermolecular interactions. Full article

Background: Excessive abdominal fat deposition is a major challenge in the chicken farming industry, making it essential to elucidate the molecular mechanisms underlying chicken adipogenesis. Nuclear Respiratory Factor 1 (NRF1) has been reported to suppress chicken adipogenesis by downregulating peroxisome proliferator-activated receptor gamma (PPARγ) expression. Protein Phosphatase 1 Catalytic Subunit Gamma (PPP1CC) is a multifunctional phosphatase involved in various biological processes; however, its role in chicken adipogenesis remains unclear. Objective: This study aimed to investigate the functional role and underlying mechanism of PPP1CC in chicken preadipocyte differentiation. Methods: Co-immunoprecipitation (Co-IP) and immunofluorescence assays were performed to determine the interaction between PPP1CC and NRF1 in DF1 cells. Bioinformatic analysis predicted potential NRF1 dephosphorylation sites targeted by PPP1CC, based on which NRF1 mutants mimicking dephosphorylation were constructed. Phos-tag SDS-PAGE combined with Western blot analysis were used to verify PPP1CC-mediated dephosphorylation of wild-type NRF1. Dual-luciferase reporter assays were used to evaluate the effect of PPP1CC-mediated dephosphorylation on NRF1-regulated PPARγ P1 promoter transcriptional activity. ChIP-qPCR was employed to assess the occupancy of NRF1 to the PPARγ P1 promoter upon PPP1CC overexpression. The effect of PPP1CC overexpression was assessed on preadipocyte differentiation using Oil Red O staining and marker gene expression analysis. Results: PPP1CC interacted with NRF1 in both the cytoplasm and nucleus of DF1 cells. Overexpression of PPP1CC significantly promoted NRF1 dephosphorylation during oleic acid-induced preadipocyte differentiation and increased endogenous NRF1 expression. Moreover, dual-luciferase assays showed that while PPP1CC strengthened the inhibitory effect of wild-type NRF1 on PPARγ P1 promoter transcriptional activity, it exerted no additional suppression on the already low activity mediated by the dephosphorylation-mimicking NRF1 mutants. Consistently, ChIP-qPCR results demonstrated that PPP1CC overexpression enhanced the occupancy of NRF1 to the PPARγ P1 promoter. Functional assays revealed that PPP1CC overexpression significantly inhibited chicken preadipocyte differentiation. Conclusions: PPP1CC interacts with NRF1 and promotes its dephosphorylation, enhancing NRF1-mediated suppression of PPARγ transcription and ultimately inhibiting chicken preadipocyte differentiation. These results identify the PPP1CC–NRF1–PPARγ regulatory axis and provide a potential molecular target for controlling fat deposition in broiler chickens. Full article

As a non-invasive mucosal immunization strategy, intranasal vaccines are highly promising for preventing respiratory infectious diseases. Among them, recombinant subunit vaccines represent a safe and ideal option, as they induce targeted mucosal immunity without the safety risks associated with live-vectored or nucleic acid vaccines. However, nasal mucosal defenses rapidly clear antigens before immune activation, limiting protective efficacy. Therefore, intranasal vaccine adjuvants—key regulators of immune response intensity, duration, and type—are essential to overcome mucosal tolerance and improve immunogenicity. Based on a systematic search and analysis of 127 peer-reviewed articles (2010–2026) in PubMed, Web of Science, and Embase, this study comprehensively summarizes the mechanisms, applications, and limitations of existing and candidate adjuvants for intranasal vaccines. This review systematically categorizes and discusses the nasal mucosal barrier, major adjuvant types (e.g., pattern recognition receptor agonists, cytokine adjuvants, and carrier adjuvants), and their mechanisms of action. It also identifies key bottlenecks: insufficient mucosal targeting, inconsistent global safety evaluation standards for adjuvants, and interference from pre-existing antibodies in humans. Furthermore, this review highlights future development directions, including biomimetic adjuvants, pH-responsive nanoadjuvants, and thermostable vaccine formulations. This systematic review clarifies key scientific and technical barriers in intranasal vaccine adjuvant development. The findings provide valuable references for advancing the translation of intranasal vaccines from emergency countermeasures to routine, accessible preventive tools for respiratory infectious diseases. Full article

The cytotoxic effect of islet amyloid polypeptide (IAPP) misfolding and aggregation has a well-recognized role in the pathogenesis of type 2 diabetes mellitus, mediated by failure of the beta cell’s protein quality control system to rescue the cell from overwhelming proteotoxic stress induced by IAPP aggregates, ultimately leading to apoptosis. A small but growing body of research also links IAPP-mediated proteotoxic stress to the pathogenesis of type 1 diabetes and to the functional decline of transplanted islets. Among the most promising therapeutic approaches under investigation are small organic molecules that may act as direct chemical chaperones to prevent IAPP aggregation, promote the activity of endogenous chaperones, or alter gene networks of the unfolded protein response (UPR) to promote pro-survival rather than pro-apoptotic pathways in response to IAPP-mediated proteotoxic stress. Compounds warranting special attention include 4-phenylbutyrate (PBA), tauroursodeoxycholic acid (TUDCA), and epigallocatechin gallate (EGCG), as each has a growing body of evidence supporting their ability to ameliorate this process, and given that each of these are already known to have good safety profiles in humans, potentially accelerating the timeline to interventional studies. This review explores the evidence for IAPP-mediated proteotoxicity in multiple forms of diabetes, the mechanisms of cytotoxicity at different levels of the cell’s protein quality control systems, how these small organic compounds may act on these processes including new insights on the role of thioredoxin-interacting protein (TXNIP), and the current evidence supporting each of these compounds in mitigating diabetogenesis. Full article

Lactic acid bacteria (LAB) are pivotal microorganisms in the food industry. Current approaches for functional gene validation and trait improvement in LAB primarily rely on traditional gene editing and homologous recombination techniques. These methods are often cumbersome, inefficient, and time-consuming, hindering the rapid and precise customization of strains. This limitation has, to some extent, constrained the rapid selection and industrial application of functional LAB strains. The engineering of LAB through gene editing technologies has significantly advanced both fundamental and applied research. Among these, CRISPR/Cas gene editing has successfully achieved precise modification of multiple genes in various LAB species. Compared to conventional methods, it offers superior editing efficiency and lower operational costs, opening new avenues for functional gene identification and genetic improvement in LAB. However, the application of exogenous CRISPR/Cas systems in LAB faces technical challenges such as high off-target rates, chromosomal abnormalities, and cytotoxicity. The development of endogenous CRISPR/Cas-based editing tools for LAB provides novel pathways for precise regulation, rational design, and flexible application. This paper first outlines the structural components and mechanistic principles of CRISPR/Cas gene editing tools. It then explores the research progress and applications of both endogenous and exogenous CRISPR/Cas systems in LAB. Finally, it provides an outlook on the future application of CRISPR/Cas gene editing technology in LAB, offering a reference for its implementation in this field. The advent of gene editing technologies has significantly propelled functional gene validation and trait improvement in lactic acid bacteria (LAB), thereby advancing both fundamental research and industrial applications. Notably, the CRISPR/Cas system has emerged as a transformative tool enabling precise genetic modification in diverse LAB species, offering marked improvements in editing efficiency and cost reduction relative to conventional approaches. CRISPR/Cas-based editing strategies in LAB are broadly classified into exogenous and endogenous systems. Exogenous systems operate independently of the host’s native immune repertoire, conferring the advantages of broad strain applicability and high editing efficiency. These systems have been successfully deployed for functional gene characterization, metabolic pathway engineering, such as augmenting antimicrobial production, and probiotic safety enhancement via virulence gene deletion. Conversely, endogenous systems leverage the intrinsic CRISPR/Cas machinery of LAB, offering superior biocompatibility and minimized off-target risks. Notable applications include precise gene knockout and integration using the native Type I-E system in Lacticaseibacillus paracasei. This review provides a concise overview of CRISPR/Cas system architecture and mechanisms, followed by a systematic synthesis of research progress and applications for both exogenous and endogenous systems in LAB. Finally, future directions are outlined to guide the continued development and application of CRISPR/Cas technologies in this field. Full article

Background: Non-invasive approaches to brain tumour detection and diagnosis are limited by the absence of clinically validated circulating biomarkers. This study utilised a miniaturised tissue perfusion model to maintain human brain tumour tissue ex vivo with the aim of identifying tissue-derived proteins with potential biomarker utility. Methods: 55 tumour samples from 11 different brain tumours (glioblastoma n = 4, low-grade glioma n = 4, brain metastases n = 3) were micro-dissected and maintained ex vivo on a continuous-flow perfusion device for 168 h. Proteomic analysis of tumour effluent was performed by reversed-phase capillary liquid chromatography-mass spectrometry. Two candidate proteins—extracellular matrix protein 1 (ECM1) and cathepsin D—were quantified using ELISA. Results: All tumour subtypes retained tissue viability over 168 h of perfusion. Proteomic profiling identified 90 tissue-derived proteins in the tumour effluent. Many proteins corresponded to previously described cancer biomarkers such as glial fibrillary acidic protein (GFAP) while others, including Serpin A12 and collapsin response mediator protein-2 (CRMP2), had not yet been described in a brain tumour context. ELISA confirmed significantly higher ECM1 levels in high-grade glioma effluent compared with low-grade glioma (p = 0.0407), whereas cathepsin D levels did not differ significantly between tumour types. Conclusions: The ex vivo perfusion model effectively preserved primary and metastatic human brain tumour tissue and enabled direct characterisation of tumour-secreted proteins. The proteins identified here warrant further validation as tumour biomarkers in patient serum or cerebrospinal fluid. Full article