Search Results (1,667)

Background/Objectives: Streptococcus pneumoniae is a major human pathogen causing illnesses that range from mild respiratory infections to severe invasive diseases. More than 100 known S. pneumoniae serotypes differ in their virulence, prevalence, and levels of drug resistance. Additionally, different clonal types within the same serotype may exhibit varying disease potential and genetic characteristics. This study aimed to determine phenotypic and molecular characteristics of S. pneumoniae isolated from patients with invasive pneumococcal disease (IPD). Methods: The serotypes of invasive S. pneumoniae isolates collected between 2022 and 2025 from adult patients hospitalized in a tertiary hospital were determined. Multilocus sequence typing (MLST) was performed on isolates with reduced susceptibility to penicillin to assess their molecular epidemiology. Results: Serotype 3 was the most common among all invasive isolates (29/85; 34.1%), followed by serotype 19A (22/85; 25.9%). Most penicillin-resistant isolates belonged to serotypes 19A and 19F. Three of the eight 19A isolates with reduced penicillin susceptibility were assigned to ST320 (37.5%), a clinically significant clone due to its high virulence and antibiotic resistance. While 15.3% of all isolates were multidrug-resistant (MDR), nearly half of the isolates with reduced penicillin susceptibility were MDR, most frequently exhibiting the erythromycin–clindamycin–tetracycline resistotype. Conclusions: This study highlights the predominance of serotype 19A, particularly the highly virulent and resistant ST320 clone, among invasive isolates with reduced penicillin susceptibility. These findings underscore the ongoing threat of antimicrobial resistance in IPD and the importance of continued surveillance of serotype distribution and resistance patterns to guide treatment strategies and vaccination policy decisions. Full article

The normal metabolism of transient starch in leaves plays a vital role in determining photosynthesis and final crop yield. However, the molecular mechanisms linking abnormal transient starch metabolism to premature leaf senescence remain unclear. Here, we isolate a rice mutant, lses1, with leaf yellowing and premature senescence, as well as excessive accumulation of starch granules in chloroplasts. Genetic analysis revealed that this trait is controlled by a single recessive nuclear gene. Through BSA-seq preliminary gene mapping, map-based cloning, and sequencing alignment, the candidate gene was pinpointed to LOC_Os02g40860 on chromosome 2, which encodes OsCKI1, a casein kinase I family member. The identity of LSES1 was confirmed functionally: genetic complementation with the native genomic sequence rescued the wild-type phenotype, while CRISPR/Cas9 knockout of the gene in wild-type plants recapitulated the premature senescence. This confirmed that LSES1/OsCKI1 is involved in the regulation of leaf senescence. Notably, one improved knockout line, KO-2, displayed significant agronomic improvements in grain length, grain width, number of productive ears, and number of filled grains per panicle, along with a significant increase in grain yield per plant, highlighting its potential breeding value. Subcellular localization and tissue-specific expression analysis showed that LSES1 is primarily nuclear-localized and constitutively expressed. Full article

Duck viral hepatitis (DVH), a highly contagious disease, is caused primarily by duck hepatitis A virus (DHAV). The viral genotypes exhibit significant diversity, creating a challenge as monovalent vaccines fail to provide cross-genotype protection in ducklings. This study aimed to design a multi-epitope peptide vaccine targeting different genotypes of DHAV. Using immunoinformatics approaches, we systematically identified key antigenic determinants, including linear B-cell epitopes, cytotoxic T-cell epitopes (CTL), and helper T-cell epitopes (HTL). Based on these, a novel vaccine candidate was developed. The vaccine construct was subjected to rigorous computational validation: (1) Molecular docking with Toll-like receptors (TLRs) predicted immune interaction potential. (2) Molecular dynamics simulations assessed complex stability. (3) In silico cloning ensured prokaryotic expression feasibility. Then, we conducted preliminary experimental validation for the actual effect of the vaccine candidate, including recombinant protein expression in E. coli, enzyme-linked immunosorbent assay (ELISA) quantification of humoral responses, and Western blot analysis of cross-reactivity. ELISA results demonstrated that the vaccine candidate could induce high-titer antibodies in immunized animals, with potency reaching up to 1:128,000, and the immune serum showed strong reactivity with recombinant VP proteins. Western blot analysis using duck sera confirmed epitope conservancy across genotypes. Collectively, the multi-epitope vaccine candidate developed in this study represents a highly promising broad-spectrum strategy against DHAV. The robust humoral immunity it elicits, coupled with its demonstrated cross-reactivity, constitutes compelling proof-of-concept, laying a solid foundation for advancing to subsequent challenge trials and translational applications. Full article

Fluconazole-resistant Candida parapsilosis has emerged as a significant nosocomial pathogen, contributing to extensive outbreaks with severe clinical implications. Despite increasing evidence of clonal transmission, the genetic mechanisms that facilitate the persistence of hospital reservoirs remain inadequately characterized. We aimed to characterise the long-term molecular epidemiology of fluconazole-resistant Candida parapsilosis bloodstream isolates (n = 47) collected between 1997 and 2019 at a tertiary centre. All isolates underwent microsatellite analysis using three polymorphic markers (CP1, CP4, B5). Genetic diversity, temporal distribution, and clonal relationships were assessed through phylogenetic analysis and discriminatory power calculations. Microsatellite analysis revealed minimal genetic diversity (combined discriminatory power: 0.7114), with only six distinct genotypes identified. Two dominant clones (Genotype-1: 23.4%, Genotype-2: 46.8%) persisted throughout the study, showing apparent spatiotemporal clustering in surgical and intensive care units. Phylogenetic analysis demonstrated tight genetic clustering, consistent with prolonged clonal persistence across multiple years and clinical departments. Our findings provide strong molecular evidence consistent with persistent, multi-year clonal transmission; however, definitive confirmation will require higher-resolution genomics and epidemiologic linkage. These results underscore the need to strengthen infection-control practices to curtail sustained clonal persistence within the hospital. Full article

Salt and low temperature are serious abiotic stresses and important constraints to agricultural productivity across the globe. These abiotic stresses negatively affect plant growth and physiological, biochemical, and molecular processes. Glycine betaine (GB) is an important osmoprotectant that enables plants to resist salinity, low temperature, and drought. GB can be synthesized in many organisms, including animals, plants, and bacteria. In higher plants, GB is synthesized through two-step oxidation of choline. However, rice, an important food crop, cannot synthesize GB. Thus, conferring the ability to synthesize GB to rice through genetic engineering is of great significance for enhancing its tolerance to abiotic stress. Recently, an enzyme, GSDMT (glycine, sarcosine, and dimethylglycine methyltransferase) was found in a diatom, Talassiosira pseudonana, and found able to catalyze the three successive methylation steps of glycine to form GB. This biosynthetic pathway for GB synthesis is also the simplest in living organisms. Here, the optimized codon of the TpGSDMT gene sequence was synthesized and cloned into an overexpression vector, pBWA(V)HS, which contains a CaMV 35S promoter, and then, the constructed vector was transferred into rice (Oryza sativa L. ssp. Japonica). The GB content in transgenic rice showing overexpression of TpGSDMT was significantly increased, and these transformants exhibited markedly enhanced tolerance to salt and low temperature. These results indicate that the TpGSDMT gene can be used for the genetic improvement in crop plants’ resistance to salinity and low temperature. Full article

Bovine parainfluenza virus type 3 (BPIV3) is a significant pathogen implicated in bovine respiratory disease complex (BRDC), leading to lung tissue destruction, immunosuppression, and subsequent bacterial infections in cattle, hence incurring considerable economic losses globally. Notwithstanding its importance, a limited number of commercial vaccinations are presently accessible. The fusion (F) protein and hemagglutinin-neuraminidase (HN) protein, as protective antigens of the Paramyxoviridae family, can elicit neutralizing antibodies and are regarded as optimal candidates for the creation of genetically modified vaccines. A multi-epitope-based peptide vaccine (MEBPV) was developed by immunoinformatics methodologies by choosing epitopes from the F and HN proteins characterized by high antigenicity, moderate toxicity, and limited allergenic potential. The epitopes were combined with suitable linkers and adjuvants to produce the vaccine, whose physicochemical qualities, immunological attributes, solubility, and structural stability were improved and evaluated using computational methods. Molecular docking and molecular dynamics simulations demonstrated the strong potential binding affinity and stability of the vaccination with TLR2, TLR3, and especially TLR4 receptors. Immune simulations forecasted strong humoral and cellular responses, accompanied by a significant elevation in interferon-γ (IFN-γ) production. The vaccine sequence was later cloned into the pET-28a (+) vector for possible expression in Escherichia coli. Despite in silico predictions suggesting a favorable immunogenic potential, additional in vitro and in vivo studies are necessary to confirm its protective efficacy and safety. This research establishes a solid foundation for the creation of safe and efficacious subunit vaccines targeting BPIV3 and presents novel perspectives for the formulation of vaccinations against additional viral infections. Full article

Aromatic coconut is a special variety of coconut. Its unique “pandan-like” aroma has won it great popularity among consumers, endowing it with considerable market potential. In our previous study, 2-acetyl-1-pyrroline (2AP), which serves as the main source of the “pandan-like” aroma in aromatic coconut, was found to exhibit significant variation among distinct aromatic coconut individuals. Now, the regulatory mechanism of 2AP has been clarified in fragrant rice, and the ProDH gene is the key gene for 2AP regulation. To further understand the regulation mechanism of 2AP content in aromatic coconut, we cloned and identified the CnProDH gene, the key gene of 2AP regulation in aromatic coconut. The results showed that the CnProDH gene had the typical ProDH structural domain, and its full-length sequence is 23,667 bp, containing 5 exons and a coding sequence (CDS) of 1599 bp. The CnProDH gene encodes a protein that possesses a β₈α₈ barrel structure, consisting of 532 amino acids (aa), with a molecular mass of 58,076.63 kDa and an isoelectric point of 7.11. To further understand the regulatory mechanism of CnProDH in aromatic coconut, we also constructed a yeast one-hybrid (Y1H) library for aromatic coconut. Through the Y1H experiment, combined with the prediction and analysis of cis-acting elements in the promoter of the CnProDH gene, three possible regulatory factors, including CnYABBY2, CnSAP8, and CnBRD3, were identified. These findings provide a molecular basis for clarifying and solving the problem of variations in 2AP content across different aromatic coconuts. Full article

Cancer stem cells (CSCs) play a crucial role in colorectal cancer by sustaining intratumoral heterogeneity, therapeutic resistance, and metastatic potential. CD133 (PROM1) is among the most frequently used surface markers for CSC identification, whereas TRIM28, a versatile epigenetic regulator, has been implicated in controlling CD133 expression and stem-like features. In this study, we performed a detailed molecular and functional analysis of Caco2 colorectal cancer cell clones with individual knockouts of CD133 or TRIM28. Elimination of CD133 neither altered global gene expression, as confirmed by transcriptome profiling, nor affected key cellular properties. In contrast, loss of TRIM28 led to a marked reduction in CD133 protein abundance and induced extensive molecular and phenotypic remodeling. TRIM28 knockout was associated with broad transcriptomic changes involving more than 500 differentially expressed genes, decreased proliferative activity monitored by time-lapse imaging, and reduced sensitivity to paclitaxel, cisplatin, and curcumin. Furthermore, immune evasion molecules CD24 and CD47 (“don’t eat me” signals) were strongly upregulated in TRIM28-deficient cells, consistently confirmed by both RNA-Seq and flow cytometry analyses. At the same time, imaging flow cytometry and mitochondrial activity assays indicated that these effects were not due to major shifts in mitotic index or bioenergetic status. Altogether, our results demonstrate that TRIM28, rather than CD133, functions as a central regulator of CSC-associated phenotypes in colorectal cancer. These findings highlight the importance of epigenetic context in CSC biology and may inform the development of more effective therapeutic strategies. Full article

Akkermansia muciniphila (AKK) is a mucin-degrading gut symbiont with emerging probiotic potential. Among its carbohydrate-active enzymes, Amuc_0517, a glycoside hydrolase family 36 (GH36) protein, has been identified as a highly specific α-galactosidase. In this study, the Amuc_0517 gene was cloned into pET-28a(+), expressed in Escherichia coli BL21, and purified via Ni²⁺-NTA affinity chromatography. Bioinformatic analysis indicated the presence of a signal peptide and α-galactosidase domain. Enzyme assays confirmed its ability to cleave α-1,6-glycosidic bonds in pNPGal, with no detectable activity toward pNPGlu, and molecular dynamics simulations revealed stronger binding affinity and lower free energy with pNPGal, supporting its substrate specificity. Given that α-galactosidases are widely applied in the dairy industry to hydrolyze galactose-containing oligosaccharides in milk and whey, the biochemical features of Amuc_0517 suggest its potential as a novel biocatalyst for functional dairy processing and probiotic-enriched dairy product development. Full article

Background/Objectives: Bacterial blight (BB) represents one of the most devastating diseases threatening global rice production. Exploring and characterizing disease resistance (R) genes provides an effective strategy for controlling BB and enhancing rice resilience. Common wild rice (Oryza rufipogon) serves as a valuable reservoir of genetic diversity and disease resistance resources. In this study, we identified and functionally characterized a novel NLR gene, YPR1, from common wild rice (Oryza rufipogon), which exhibited significant spatial, temporal, and tissue-specific expression patterns. Methods: Using a combination of conventional PCR, RT-PCR, bioinformatics, transgenic analysis, and CRISPR/Cas9 gene-editing approaches, the full-length YPR1 sequence was successfully cloned. Results: The gene spans 4689 bp with a coding sequence (CDS) of 2979 bp, encoding a 992-amino acid protein. Protein domain prediction revealed that YPR1 is a typical CNL-type NLR protein, comprising RX-CC_like, NB-ARC, and LRR domains. The predicted molecular weight of the protein is 112.43 kDa, and the theoretical isoelectric point (pI) is 8.36. The absence of both signal peptide and transmembrane domains suggests that YPR1 functions intracellularly. Furthermore, the presence of multiple phosphorylation sites across diverse residues implies a potential role for post-translational regulation in its signal transduction function. Sequence alignment showed that YPR1 shared 94.02% similarity with Os09g34160 and up to 96.47% identity with its closest homolog in the NCBI database, confirming that YPR1 is a previously unreported gene. To verify its role in disease resistance, an overexpression vector (Ubi–YPR1) was constructed and introduced into the BB-susceptible rice cultivar JG30 via Agrobacterium tumefaciens-mediated transformation. T₁ transgenic lines were subsequently inoculated with 15 highly virulent Xanthomonas oryzae pv. oryzae (Xoo) strains. The transgenic plants exhibited strong resistance to eight strains (YM1, YM187, C1, C5, C6, T7147, PB, and HZhj19), demonstrating a broad-spectrum resistance pattern. Conversely, CRISPR/Cas9-mediated knockout of YPR1 in common wild rice resulted in increased susceptibility to most Xoo strains. Although the resistance of knockout lines to strains C7 and YM187 was comparable to that of the wild type (YP^WT), the majority of knockout plants exhibited more severe symptoms and significantly lower YPR1 expression levels compared with YP^WT. Conclusions: Collectively, these findings demonstrate that YPR1 plays a crucial role in bacterial blight resistance in common wild rice. As a novel CNL-type NLR gene conferring specific resistance to multiple Xoo strains, YPR1 provides a promising genetic resource for the molecular breeding of BB-resistant rice varieties. Full article

In this study, we successfully cloned the fluorescent proteins eGFP and DsRed in-frame with the antimicrobial peptide epinecidin-1 (FIFHIIKGLFHAGKMIHGLV) at the N-terminal. The cloning strategy involved inserting the fluorescent reporters into the expression vector, followed by screening for positive clones through visual fluorescence detection and molecular validation. The visually identified fluorescent colonies were confirmed as positive by PCR and plasmid migration assays, indicating successful cloning. This fusion of fluorescent reporters with a short antimicrobial peptide enables real-time visualization and monitoring of the peptide’s mechanism of action on membranes and within cells, both in vivo and in vitro. The fusion of eGFP and DsRed to epinecidin-1 did not impair the expression or fluorescence of the reporter protein. Full article

Background/Objectives: Streptococcus suis is a zoonotic pathogen that causes infections in pigs and humans, leading to significant economic losses. S. suis can evade the immune system of hosts and induce persistent infections. Early detection and vaccination are crucial for controlling the disease in swine industries. This study aimed to investigate candidate recombinant protein for antibodies against S. suis detection and subunit vaccine development. Methods: The whole genome of S. suis BM407 was analyzed using bioinformatic tools to predict suitable proteins and genes for recombinant protein expression. Partial extracellular factor protein (epf) genes of S. suis serotype 2 DMST18783 were amplified. A 3301 bp amplicon was digested, and a specific 615 bp fragment was inserted into a pQE81L-KAN vector. Then, the constructed plasmid was cloned and expressed in Escherichia coli DH10β. Purified protein was analyzed using SDS-PAGE. In addition, translated amino acid sequences were analyzed for immune response properties, molecular docking, molecular dynamic simulation, and epitope prediction. Results: The amino acid sequence of recombinant extracellular factor protein (rEF) was revealed as a promising antigen containing putative protective regions as linear epitopes. Furthermore, the rEF was expressed as a histidine-tagged recombinant protein, and its properties were nearly similar to the predicted rEF using bioinformatic tools. Binding of the recombinant EF (rEF) protein was found to reduce fluctuations in the swine toll-like receptor 2. Furthermore, the rEF contained several regions that were predicted to be epitopes for both B-cells and T-cells. Conclusions: This study indicates that the recombinant EF fragment is a promising candidate for detecting antibodies against S. suis and as a component of a subunit vaccine. Full article

Anastatus orientalis, a prominent egg parasitoid of Lycorma delicatula, demonstrates considerable potential for biological control. A. orientalis is dependent on host volatiles to identify and locate appropriate hosts for reproduction, with its olfactory system playing a vital role in volatile detection. There is little known about the chemosensory genes in A. orientalis. Therefore, here, we conducted a transcriptome analysis of the males and females from A. orientalis. Overall, 24 odorant binding proteins (OBPs), 4 chemosensory proteins (CSPs), 26 odorant receptors (ORs), 3 gustatory receptors (GRs), 3 ionotropic receptors (IRs), and 2 sensory neuron membrane proteins (SNMPs) were identified by transcriptome analysis. The values for fragments per kilobase per million (FPKM) indicated that the chemosensory protein gene families in A. orientalis exhibit different expression levels in male and female adults, with some genes showing significant differences and displaying sex-biased expression. Furthermore, RACE technology, phylogenetic analysis, and expression analysis were used to investigate the role that AoOBP8 plays in olfaction in A. orientalis. AoOBP8 was highly expressed in females and the heads of adults, indicating that the gene has a crucial role to play in the search for hosts and in oviposition in female adults, while the head is crucial in recognizing chemical information. These results contribute to a deeper understanding of the functions of chemosensory protein gene families in A. orientalis and offer a reference for developing biocontrol methods for forestry pests. Full article

The use of insects for feed has a significant impact on aquaculture, contributing to the achievement of the Sustainable Development Goal of Zero Hunger and Sustainable Agriculture (SDG 2), among others. This study mapped the intermediate Technology Readiness Levels (TRLs), encompassing scientific knowledge (TRL 3) through 971 scientific articles (Scopus) and technological development (TRLs 4–5) through 218 patents (Espacenet). The highest conversions from TRL 3 to TRLs 4–5 were observed for fish, mollusks, crustaceans, and annelids. Key technological targets include carp and black soldier flies (BSF). Most technologies follow circular economy principles. Emerging themes include immunity, cloning, molecular techniques, metabolomics, and genetics. China leads in TRLs 3–5, followed by the United States. Only France, the United States, and five additional countries hold export-oriented patents targeting 26 markets, primarily involving BSF-based feed formulations. Future growth trends are exponential for scientific articles, logarithmic for total patents, and linear for export patents. Collaboration at TRLs 4–5 remains limited, underscoring the need for greater international cooperation to expand access to sustainable insect-based aquaculture feed technologies. Full article

Chilli veinal mottle virus (ChiVMV) causes severe yield losses in pepper across Asia. It is very urgent to study the host plant resistance to control this viral disease. As a type of defense response gene, pathogenesis-related protein 1 (PR1) is a well-established defense marker against fungal/bacterial pathogens, and its role in virus resistance remains unclear. Here, we cloned CaPR1 from the ChiVMV-highly resistant pepper variety ‘Perennial’. The 477 bp ORF encodes a 17.65 kDa basic protein containing a conserved CAP-PR1 domain. The subcellular localization of CaPR1 revealed that it was located in the plasma membrane, endoplasmic reticulum (ER), and nucleus. RT-qPCR revealed leaf-predominant expression, with earlier and stronger induction in the highly resistant than the highly susceptible variety after ChiVMV inoculation (6.4-fold at 2 days post-inoculation). The overexpression of CaPR1 in tobacco significantly increased resistance, reducing disease index by 25% and viral coat protein accumulation. Our findings identified CaPR1 as a positive regulator of ChiVMV resistance, providing a molecular target for pepper breeding. In addition, exogenous SA treatment increased the resistance of the highly susceptible cultivar ‘Guijiao 12’ to ChiVMV, and 0.25 mM had a greater effect. Full article