Search Results (9,027)

Purified water from vending machines offers consumers an alternative source of clean, safe water. However, data regarding its microbiological quality are limited, particularly concerning the prevalence of Pseudomonas aeruginosa harboring virulence traits. This study aimed to evaluate the microbiological quality of 125 purified water samples collected from vending machines across six cities of Michoacan, Mexico. Additionally, it sought to assess the occurrence of Pseudomonas aeruginosa and characterize its antimicrobial resistance profiles and biofilm-forming capacity. Aerobic mesophilic bacteria (AMB) were detected in all analyzed samples. A total of 71 (56.8%), 40 (32.0%), and 31 (24.8%) samples were positive for total coliforms (TC), fecal coliforms (FC), and Escherichia coli, respectively. Among the samples, 43 (34.4%) were positive for P. aeruginosa. There were significant correlations between the presence of P. aeruginosa and AMB (rho = 0.4445; p < 0.0001), TC (rho = 0.4094; p < 0.0001), FC (rho = 0.3389; p = 0.0001), and E. coli (rho = 0.3242; p = 0.0002). Moreover, the presence of TC in purified water samples increased the risk of P. aeruginosa nearly seven-fold (odds ratio = 6.91; p < 0.001). The resistance rate among P. aeruginosa strains to the most tested antibiotics ranged from 2.3 to 16.3%, and two (4.6%) of the isolates were multidrug-resistant. All P. aeruginosa strains were strong biofilm producers. Consequently, we recommend periodic maintenance of vending machines, the establishment of P. aeruginosa control protocols, and enhanced regulatory monitoring of the water vending industry. Full article

The plant-specific TCP transcription factor family plays crucial roles in morphogenesis and stress adaptation. While characterized in many species, this family remains unstudied in Punica granatum. We performed the first genome-wide analysis of the TCP family in pomegranate, identifying 24 PgTCP genes classified into the PCF, CIN, and CYC/TB1 subclades, supported by conserved gene structures and motifs. Evolutionary analysis indicated segmental duplication and purifying selection shaped this family. Expression profiling revealed distinct spatiotemporal patterns: PgTCP2/9/14/21 were highly expressed in flowers, with PgTCP21 also notably abundant in fruit tissues (seed coats and pericarp), suggesting roles in reproductive development. PgTCP19, an ortholog of the branching suppressor BRC1, showed dominant expression in dormant buds, implicating it in shoot architecture regulation. Furthermore, PgTCP5 and the miR319-targeted PgTCP22 were leaf-predominant, indicating a function in leaf development. Under abiotic stress, PgTCPs displayed dynamic, treatment-specific responses. A subset of genes was rapidly induced by cold, while PgTCP14 and PgTCP23 showed sustained upregulation during drought. Several light-responsive PgTCPs were suppressed under shading. This study provides a foundational resource, functionally classifies the PgTCP family, and identifies key candidates regulating organ development and stress resilience for future functional validation and molecular breeding in pomegranate. This work provides the first comprehensive overview of the TCP gene family in pomegranate and offers candidate genes for future functional studies related to development and stress responses. Full article

Pinoresinol diglucoside (PDG), one of the major lignans isolated from E. ulmoides Oliver bark, has various pharmacological functions, including antihypertension and prevention of osteoporosis. However, the glycosyltransferase-encoding gene (GT) involved in regulating the glycosylation of pinoresinol to form PDG has not been reported in E. ulmoides. In this study, we screened and cloned the EuGT8 gene from E. ulmoides based on our transcriptome data. The expression pattern of the EuGT8 gene exhibited a strong positive correlation with dynamic changes in the PDG contents in three different organs of E. ulmoides. The expression level of the EuGT8 gene and PDG content were significantly decreased in asODN-EuGT8-treated shoot tips in comparison with the control group. Prokaryotic expression of the EuGT8 gene revealed that the purified EuGT8 protein could catalyze the conversion of pinoresinol into PDG. In addition, we performed transcriptional and metabolomic analyses to compare the differences between transgenic Arabidopsis and WT plants. A total of 1799 DEGs and 294 DEMs were identified in transgenic and WT plants. KEGG enrichment analysis showed that the DEGs were mainly enriched in phenylpropanoid biosynthesis, secondary metabolite biosynthesis, and starch/sucrose metabolism pathways. The DEMs were mainly enriched in ABC transporters, aminoacyl-tRNA biosynthesis, biosynthesis of amino acids, phenylpropanoid biosynthesis, and flavone and flavonol biosynthesis pathways. Correlation analysis between DEGs and DEMs identified a total of 231 DEGs associated with 38 DEMs, which were mainly distributed in multiple metabolic pathways. This finding provides both theoretical insights and genetic resources for breeding high-PDG E. ulmoides varieties, facilitating marker-assisted selection (MAS) and promoting sustainable E. ulmoides production in Guizhou. Full article

We develop an empirical application on a large dataset of European stock returns in order to estimate the risk premia. While traditional factor models often struggle with high levels of pricing errors and noisy proxies in fragmented markets, we show that the Three-Pass Estimation Method (3PEM) serves as both a robust estimator and a diagnostic tool for factor purification. By assuming the Fama–French five-factor model as the baseline model, we first show that the 3PEM yields risk premium estimates for the European market that are more economically plausible and statistically robust than those obtained using the traditional two-pass estimation method (2PEM). Moreover, our results show that the 3PEM is able to detect noise in tradable factors. Furthermore, the 3PEM is used to denoise the observed factors, providing purified versions that better capture the systematic components of risk. We also identify both noisy factors and denoised factor series that improve the estimation of stock-level exposures and expected returns. Full article

Background: Human cysticercosis, caused by the larval stage (cysticerci) of the pork tapeworm Taenia solium, is an important zoonotic disease. The disease is prevalent in developing countries where porcine cysticercosis is common and undercooked pork is habitually consumed. Objective: This study aimed to develop an immunochromatography-based test kit for the rapid diagnosis of human cysticercosis using low-molecular-weight antigens purified from cyst fluid of the T. solium Asian genotype to detect specific IgG antibodies in whole blood. The kit was designated as “the cysticercosis whole-blood test kit (iCys WB kit).” Methods: It was evaluated under laboratory conditions using 164 whole-blood samples, of which 21 were from confirmed cysticercosis cases. The results of the iCys WB kit, which detects anti-T. solium (cysticercus) antibodies in simulated whole blood samples, were compared with results from corresponding human serum samples. Results: When using both sample types, iCys WB kit demonstrated an accuracy of 98.8%, a sensitivity of 91.7%, a specificity of 100%, a positive likelihood ratio of 0, a negative likelihood ratio of 0.083, and an ROC area of 0.96. The agreement between results obtained from simulated whole-blood and serum samples showed perfect concordance. Conclusions: The iCys WB kit is a valuable easy-to-handle diagnostic tool and may be applicable for supporting clinical diagnosis at the point of care. Full article

Background: Mycoplasma pneumoniae (MP) is a major cause of respiratory tract infections in children and adolescents. Currently, there is no licensed vaccine, underscoring the urgent need for the development of safe and effective vaccines. Objective: The aim of this study is to develop a recombinant subunit vaccine candidate incorporating three antigens: the P1 protein, the P40/90 complex, and a detoxified mutant of community-acquired respiratory distress syndrome toxin. The protective efficacy of this vaccine candidate was also evaluated. Methods: Target genes were codon-optimized for expression in E. coli, and the recombinant proteins were successfully expressed and purified. The low-toxicity CARDS toxin mutant was screened based on TNF-α secretion levels in stimulated RAW264.7 cells. A three-component vaccine composed of P1, P40/90, and the mutant CARDS toxin was formulated and adjuvanted with either Al(OH)₃ alone or in combination with CpG. Mice were immunized, and immunogenicity was assessed by measuring antigen-specific IgG antibody titers. Protective efficacy was evaluated following challenge by analyzing lung histopathology, bacterial load, and inflammatory cytokine levels. Results: Seven high-purity recombinant proteins were successfully produced, including P1, the P40/90 complex, wild-type CARDS toxin, and four CARDS toxin mutants (E132A, E132Q, H36A, R10A). The E132A mutant was selected due to its significantly reduced toxicity while retaining immunogenicity. The three-component vaccine effectively elicited antibody responses against each of the included antigens. After three immunizations, IgG antibody titers in all groups reached approximately 10⁴. Immunized mice showed markedly reduced pulmonary pathology scores (control group: 2 or 2.67; immunized groups: 1.67, 1.33, and 0) and significantly decreased bacterial loads in lung tissue (control: 30.11 ± 10.40 cp/μL; immunized groups: 20.72 ± 4.37 cp/μL and 8.51 ± 8.32 cp/μL). Furthermore, the group receiving the alum + CpG adjuvant exhibited approximately a 10-fold higher antibody response compared with the alum-only group, indicating enhanced protective efficacy. Conclusions: The three-component candidate vaccine, MPtriV, adjuvanted with Al(OH)₃ + CpG, demonstrates promising immunogenicity, safety, and protective efficacy against Mycoplasma pneumoniae infection, providing a viable strategy and experimental foundation for the development of MP subunit vaccines. Full article

House dust mite allergens Der p 1, Der p 2, and Der p 23 are recognized as major clinically relevant allergens worldwide; however, it is difficult to obtain these proteins in purified form from a natural source, which limits their use in molecular targeted immunotherapy and in vivo diagnosis. In this study, we developed and validated robust methodologies for the large-scale purification and individual characterization of native nDer p 1, nDer p 2, and nDer p 23 allergens from the natural sensitization source, Dermatophagoides pteronyssinus. Each allergen was isolated through an independent downstream process based on successive chromatographic steps, achieving high purity and preserving the structural integrity. Molecular standardization was performed in vivo in 27 mite-allergic patients by skin prick testing (SPT), enabling the separate determination of histamine equivalent potency (HEP) values: 7.43 µg/mL for nDer p 1, 8.11 µg/mL for nDer p 2, and 1.55 µg/mL for nDer p 23. These data establish a direct relationship between the protein concentration and biological activity for each major allergen. In conclusion, the successful production and biological standardization of native nDer p 1, nDer p 2, and nDer p 23 proteins provide well-defined reagents for in vivo molecular diagnosis and enable more precise and reproducible standardization compared with complex allergen extracts. Full article

Aerosols are a major transmission route for seasonal influenza infections. Titanium dioxide (TiO₂) photocatalyst has broad-spectrum antiviral activity, including in vitro influenza virus inactivation; however, whether the TiO₂ photocatalyst can effectively inactivate airborne influenza A viruses in vivo under conditions that mimic natural aerosol transmission remains unclear. Here, we evaluated in vivo inactivation of airborne H1N1 seasonal influenza virus by a photocatalyst-equipped air purifier using a mouse model. Influenza virus WSN strain aerosols were sprayed in a 60 L acrylic box with a nebulizer, circulated through a photocatalyst-equipped air purifier, exposed to BALB/c mice for 40 min after circulation, and subsequently collected with an air sampler. Thirty minutes of TiO₂ photocatalyst treatment reduced influenza virus infectivity by 99.97%, and significantly lowered lung viral titer in mice on day 3 post-infection. Over 14 days post-infection, mice showed no >10% weight loss, 100% survival, and disease progression to the PBS (−) aerosol group. This suggests that the photocatalyst-equipped air purifier may reduce H1N1 seasonal influenza onset, preventing viral spread. Full article

An abnormal increase in acne-causing bacteria is the main cause of acne. This study aimed to investigate Piper griffithii C.DC. as a new source of compounds for inhibiting acne-causing bacteria and to provide the first elucidation of the mechanism of action against these bacteria. The antibacterial efficacy of 27 Piper species was examined against acne-causing clindamycin-resistant bacterial strains. Antibacterial activity of various crude extracts derived from leaves or stems extracted using hexane, ethyl acetate, or ethanol was evaluated. Ethyl acetate leaf extract of P. griffithii exhibited the greatest antibacterial effect against all tested bacteria. Zeylenone, an antibacterial substance isolated, purified, and characterized from the ethyl acetate leaf extract of P. griffithii, disrupts cell walls and membranes. Minimum bactericidal concentration (MBC) values were 1.25, 2.5, and 7.5 mg/mL for Cutibacterium acnes, Staphylococcus aureus, and S. epidermidis, respectively. Zeylenone derived from P. griffithii leaves was nontoxic to human skin keratinocytes (HaCaT). A formulated anti-acne gel with zeylenone was effective in controlling acne-causing bacteria. These results suggest that zeylenone isolated from P. griffithii leaves can be further developed as a natural ingredient in anti-acne products. This is the first report of the use of zeylenone from P. griffithii for eliminating acne-causing bacteria. Full article

Pectate lyase (PeL) is a key cell wall-degrading enzyme in the infection process of plant-parasitic nematodes, with a large gene family exhibiting functional redundancy. The dominant PeL isoform during the initial infection time course remains unclear. In this study, 21 Ddpel genes were identified in Ditylenchus destructor Thorne, 1945, 7 of which were differentially expressed during the initial infection time course of this nematode. The purified proteins of these seven DdPeLs showed pathogenicity toward both sweet potato and tobacco, and their optimal enzymatic pH varied significantly. Prior to host infection, D. destructor preferentially expresses Ddpel genes encoding pectate lyase with higher activity at pH 5.8. However, within 5 days post-inoculation with nematodes, the expression of genes encoding acidic DdPeL enzymes (enzymes with optimal activity in acidic pH) was upregulated, while genes encoding alkaline DdPeL enzymes (optimal activity in alkaline pH) were concurrently downregulated. Through site-directed mutagenesis, we demonstrated that the loss of enzymatic activity in DdPeLs abolished their ability to induce plant cell death. Furthermore, when acidic or alkaline DdPeLs were pre-treated with dialysis in their respective optimal pH buffers prior to infiltration, their pathogenicity was significantly enhanced. Together, these findings demonstrate that enzymatic activity, governed by protein structure and local pH, is a key determinant of pathogenicity. Previous studies have reported that phytopathogens can secrete organic acids during the initial infection phase, leading to localized acidification of the host microenvironment. We therefore hypothesize that, during the initial infection time course, nematodes may actively acidify the host microenvironment to specifically enhance the enzymatic activity of acidic DdPeLs, thereby promoting cell wall degradation and facilitating infection establishment. Full article

Background: Rabies is a highly fatal zoonotic disease that causes approximately 59,000 human deaths worldwide each year. Current inactivated rabies vaccines require multiple doses and are associated with high costs. The full-length rabies virus glycoprotein (RVG), a membrane protein, exhibits substantial instability in its trimeric structure during recombinant expression. This instability makes it difficult to obtain high-purity, correctly folded antigens. Objectives: This study focuses on the preparation of a full-length recombinant RVG subunit vaccine candidate expressed in a glycoengineered Pichia pastoris system with mammalian-like glycosylation. Methods: The full-length RVG gene (including the transmembrane domain and cytoplasmic tail) from the Challenge Virus Standard-11 (CVS-11) strain was codon-optimized and inserted into the pPICZαA vector to construct the recombinant expression plasmid pPICZαA-RVG. The plasmid was transformed into glycoengineered Pichia pastoris X33-7 (low-mannose type) by electroporation for inducible expression. The target protein was purified by nickel affinity chromatography, anion-exchange chromatography, and Superdex-200 size-exclusion chromatography. The structural characteristics of the purified protein were analyzed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). The purified antigen was formulated with the adjuvants AS03 or MF59. BALB/c mice (n = 5 per group) were immunized intramuscularly following a four-dose schedule (days 0, 7, 14, and 28). Antigen-specific IgG antibody titers were measured by ELISA, and neutralizing antibody titers were determined using the rapid fluorescent focus inhibition test (RFFIT). Results: Glycoengineered Pichia pastoris yeast strains expressing wild-type RVG (RVG-WT) or a mutant variant (RVG-M6: R84S, R199S, H270P, R279S, K300S, and R463S) were successfully constructed. The purified RVG antigen formed nanoparticles with an average particle size of approximately 75 nm. Immunized mice generated robust RVG-specific IgG responses, with titers reaching approximately 6.31 × 10⁵ for RVG-WT after the fourth immunization, compared to 3.16 × 10³ for RVG-M6 and 5.62 × 10³ for the RVG-WT-PEG control. Two weeks after the fourth immunization, RVG-WT formulated with AS03 or MF59 induced significant neutralizing antibody responses compared with the control group (p < 0.0001 and p < 0.01, respectively). The neutralizing antibody titers reached 1:79.43 in the AS03 group and 1:33.11 in the MF59 group, whereas the WT-PEG + AS03 control group showed a low titer of 1:3.72. In contrast, RVG-M6 formulated with MF59 failed to induce detectable neutralizing antibodies (1:3.02). Furthermore, RVG-WT + AS03 induced significantly higher neutralizing antibody responses than the WT-PEG + AS03 control group (p < 0.0001), and a significant difference was also observed between the RVG-WT + MF59 and RVG-M6 + MF59 groups (p < 0.01). Conclusions: The glycoengineered Pichia pastoris expression system successfully produced uniform full-length rabies virus glycoprotein nanoparticles with high purity. When formulated with the AS03 adjuvant, RVG-WT induced high-titer neutralizing antibodies in mice, suggesting a promising strategy for the development of recombinant subunit vaccines against rabies. However, this study is limited by the absence of challenge studies and validation in target animal species, which will be further investigated in future work. Full article

Biological manipulation via physical stimuli such as light and magnetism has become a central goal in modern biotechnology. Among these modalities, magnetic fields offer unique advantages, including deep tissue penetration and untethered interventions in living systems. An ideal platform for such a magnetogenetic toolkit would be a genetically encodable protein with tunable magnetic features under physiological conditions. However, the development of such tools has been hindered by the lack of robust and stable protein scaffolds with strong intrinsic magnetic properties. Inspired by animal magnetoreception in nature, here, we rationally designed and systematically screened single-chain variants of the magnetoreceptor MagR. Through nine iterative rounds of design and experimental validation, we generated 25 constructs and ultimately identified a stable single-chain-dimer-based-tetramer, SDT-MagR, as the optimal magnetic molecular platform. This engineered protein exhibits exceptional structural stability and state-dependent magnetic behavior, showing ferrimagnetic-like characteristics in the solid state and paramagnetic behavior in solution. With enhanced magnetic susceptibility, purified SDT-MagR can be directly attracted by a magnet in vitro, establishing it as a promising new platform for future biomagnetic manipulation and magnetogenetics applications. Full article

Background and Objectives: We aimed to evaluate differences in PM exposure reduction behaviors among older adults and patients with chronic respiratory diseases, atrial fibrillation, and stroke and to identify associated determinants. Materials and Methods: This multicenter cross-sectional study included 717 participants from five cohorts: older adults (n = 255), chronic obstructive pulmonary disease (COPD, n = 145), asthma (n = 100), atrial fibrillation (n = 117), and stroke (n = 100). PM exposure reduction behaviors were assessed using a standardized 18-item questionnaire (total score range: 0–126) covering indoor, outdoor, and other behaviors (health-seeking actions, such as checking air quality and using purifiers). Results: In multivariable linear regression models, COPD (β = 11.09), asthma (β = 15.54), and atrial fibrillation (β = 14.29) were associated with significantly higher total behavior scores compared with the older adult cohort (adjusted mean 69.0), corresponding to an approximately 20% relative increase in the asthma cohort. The stroke cohort did not differ significantly from the primary cohort. Domain-specific analyses showed that indoor and other behavioral scores were consistently higher across all disease cohorts, whereas outdoor scores were significantly elevated only for asthma and atrial fibrillation. In a fully adjusted model that included all covariates simultaneously, the stroke cohort demonstrated a modest increase in the total score (β = 8.79). Increased age and higher educational attainment were independently associated with greater behavioral engagement. Conclusions: PM exposure reduction behavior scores differed across the clinical cohorts, and sociodemographic factors were independently associated with behavioral engagement. These findings support personalized disease-specific counseling and inform future digital health interventions for vulnerable populations. Full article

Codon usage bias (CUB) is a universal feature in both mitochondrial and ribosomal genes, shaped by the combined forces of mutation and selection, and serves as a valuable indicator of evolutionary processes and phylogenetic signals. However, comprehensive analysis of CUB is lacking in the Leuciscinae family. We sequenced and annotated the complete mitochondrial genome of Abramis brama orientalis and examined codon usage patterns in all Leuciscinae species, subsequently finding the dominant evolutionary forces and phylogenetic relationships. We performed a PR2 bias plot, neutrality plot, effective number of codons (ENC) vs. GC3, relative synonymous codon use (RSCU) clustering and Ka/Ks calculations for 22 Leuciscinae species. We constructed the phylogenetic trees of Leuciscinae using Bayesian inference and maximum likelihood on concatenated mitochondrial sequences. The complete mitochondrial genome of Abramis brama orientalis was 16,607 bp, with typical vertebrate structure and high A + T bias. The codons of protein-coding genes in Leuciscinae have a preference for ending in A/C. All protein-coding genes were under strong purifying selection (Ka/Ks < 1). RSCU patterns and phylogenetic analyses showed two lines of Leuciscinae in the RSCU, with A. brama orientalis being a monophyletic group with A. brama. The results demonstrate the strong role for selection in shaping mitochondrial codon usage in Leuciscinae, despite mutational biases. The study clarified the taxonomic status of A. brama orientalis and provided a framework for understanding molecular evolution in this ecologically important freshwater fish family. Full article

Energy transformation requires the development of distributed renewable energy, in which heat and electricity are produced by small units or production facilities for local needs. One favorable development direction is the thermal conversion of biomass, which is classified as a renewable energy source. Due to the variability of its physicochemical properties, gasification technology offers a flexible and competitive alternative to combustion processes. One of the key challenges associated with biomass gasification is the relatively high concentration of contaminants in the raw producer gas. This article presents the results of pilot studies on producer gas purification using activated carbon fixed-bed adsorption. The pilot studies focused on assessing the effectiveness of this technology in the context of purifying producer gas from biomass gasification installations. During the conducted experimental study, approximately 2.2 kg of contaminants were adsorbed. The calculated unit mass of adsorbed contaminants per unit volume of producer gas was 11.7 g/Nm³. The removal efficiency of contaminants was 61.5% for tar compounds and 83.6% for volatile organic compounds. A 100% removal efficiency was achieved for the analyzed sulfur compounds (H₂S, COS, and CH₃SH). The research showed positive effects of adsorption for final producer gas purification, supporting further experimental research. Full article