Search Results (293)

Trehalose is a functional disaccharide widely used in the food, pharmaceutical, and cosmetic industries. It is industrially produced via a dual-enzyme process involving maltoligosaccharide trehalose synthase (MTSase) and maltoligosaccharide trehalose hydrolase (MTHase), with Escherichia coli (E. coli) serving as the expression host. Bacillus subtilis (B. subtilis) is an ideal host for industrial trehalose production due to its generally recognized as safe (GRAS) status and low phage susceptibility. However, engineered B. subtilis strains often exhibit slow growth, low heterologous protein expression, and high fermentation costs, thereby limiting their industrial application. To address these challenges, this study employed a synergistic strategy that combined chassis modification, expression element optimization, and knockout of substrate-competition pathways. First, a tryptophan-independent strain was constructed by reverting the trpC2 mutation to shorten the growth cycle. Next, knockout of flgD, yueB, and integration of E. coli-derived glutamate dehydrogenase (gdhA) significantly enhanced biomass accumulation. Expression of MTSase and MTHase was markedly improved through tandem strong promoters (P_HpaII-P₃₆) and ribosome-binding site (RBS) optimization (RBS1), achieving a 10.87-fold and 4.22-fold increase in enzyme activity, respectively. Finally, disruption of the amyE gene reduced non-specific substrate degradation. Using maltodextrin as substrate, the final trehalose conversion rate reached 76%. This study constructed B. subtilis chassis cells that highly express MTHase and MTSase respectively, laying a foundation for subsequent industrial trehalose production. Full article

Probiotic strains with multifunctional skin-beneficial properties represent promising candidates for next-generation cosmetic ingredients. In this study, a systematic stepwise screening strategy was applied to an in-house bacterial library comprising 302 isolates to identify candidates with antioxidant, anti-inflammatory, antimicrobial, wound-healing, and skin barrier-supportive activities. From this library, 33 strains were selected based on preliminary assessments and subjected to comprehensive in vitro evaluation. Cell viability and cytotoxicity assays confirmed the safety of all selected strains in RAW264.7 macrophages and HaCaT keratinocytes. Several strains exhibited strong DPPH radical scavenging activity and significantly inhibited nitric oxide production in LPS-stimulated macrophages. Among the selected candidates, Lacticaseibacillus rhamnosus DM073 demonstrated the most potent anti-inflammatory activity, whereas Lactiplantibacillus plantarum DM043 exhibited the greatest wound-healing capacity. All five selected strains displayed antimicrobial activity against Cutibacterium acnes. Furthermore, selected strains, particularly Lactiplantibacillus plantarum DM175 and Ligilactobacillus salivarius DM079, enhanced the expression of skin barrier-related genes, including zonula occludens-1 (ZO-1), occludin (OCLN), claudin-1 (Cla-1), and filaggrin (FLG), and partially restored their expression under TNF-α/IFN-γ-induced inflammatory conditions. Selected strains also reduced the expression of inflammatory chemokines in stimulated keratinocytes. Collectively, these findings demonstrate that the selected probiotic strains possess complementary multifunctional activities associated with skin health. In particular, DM073 exhibited superior anti-inflammatory activity, DM043 showed strong wound-healing potential, and DM175 demonstrated remarkable skin barrier-supportive effects. These strain-specific properties support their potential application in the development of probiotic-derived postbiotic cosmetic ingredients for skin soothing, barrier reinforcement, skin recovery, and microbiome-friendly skincare formulations. Further studies are warranted to evaluate their efficacy and safety in advanced skin models and clinical cosmetic applications. Full article

This study investigates the enhancement of electrical conductivity in poly(3,4-ethylenedioxythiophene):polystyrene sulfonate (PEDOT:PSS) thin films through the incorporation of few-layer graphene (mG). Nanocomposite films were prepared by spin coating from liquid dispersions containing approximately 10 wt% mG. The resulting films exhibited high optical transmittance (~80%) and significantly reduced sheet resistance, reaching values as low as 1.8 kΩ/□. These improvements in electrical and optical performance are attributed to enhanced charge transport arising from π–π interactions between graphene and PEDOT:PSS, as well as conformational changes in the polymer chains. The PEDOT:PSS/mG composites were successfully applied both as conductive inks, forming conductive trails capable of powering a light-emitting diode (LED), and as hole transport layers in organic light-emitting diodes (OLEDs). Comprehensive optical and electrical characterization of the composite films and the corresponding OLED devices demonstrates the strong potential of PEDOT:PSS/mG nanocomposites for use in flexible and printed electronic applications. Full article

Industrial gear contacts operate under mixed-to-boundary lubrication where reliable antiwear protection is essential. This study assesses whether carbon nanomaterials can enhance the performance of zinc dialkyldithiophosphate (ZDDP) under severe conditions. A crossed-cylinder Reichert configuration (2 GPa, 75 °C, 1 m/s) with PAO6 was used to test ZDDP (1 wt%) and its blends with carbon nanotubes (CNT, 0.05 wt%), graphene nanoplatelets (GNP, 0.05 wt%), and few-layer graphene (FLG, 0.05 wt%) at 1, 10 and 60 min. The lubrication regime was boundary. Friction, specific wear rate (k), and tribofilm coverage were quantified. Oils containing only carbon nanoparticles could not sustain the test (seizure within minutes), confirming the necessity of ZDDP. After 60 min, average CoF remained similar across formulations and largely governed by ZDDP. By contrast, wear showed marked differences: relative to ZDDP alone (A), ZDDP + CNT (F) and ZDDP + GNP (G) reduced k by 52% and 48%, respectively, and exhibited higher tribofilm coverage (F = 68%, G = 72% vs. A = 57%). Time-resolved tests revealed that long-duration degradation was mitigated in F and G: from 10 to 60 min, k rose by 72% (F) and 58% (G) versus 159% for A; coverage decreased by only 8% (F) and 3% (G) versus 22% for A. SEM–EDS indicated no major differences in average elemental chemistry among formulations, suggesting an improvement on tribofilm coverage/stability rather than compositional change. Full article

Background/Objectives: Vitamin D deficiency is a global health concern, yet circulating 25-hydroxyvitamin D (25OHD) concentrations vary substantially across geographical regions and ancestral groups. Genetic predisposition may contribute to these differences. This systematic review aimed to synthesize evidence from genome-wide association studies (GWAS) on genetic variation associated with circulating 25OHD across populations from different ancestral backgrounds and to evaluate linkage disequilibrium (LD) between reported variants. Methods: A systematic review was conducted according to PRISMA 2020 guidelines. PubMed and the GWAS Catalog were searched to identify genome-wide association studies (GWAS) on circulating 25-hydroxyvitamin D (25OHD) concentrations. Studies were screened against predefined eligibility criteria, and data were extracted using a standardized framework. Methodological quality was assessed using a standardized tool, and study power adequacy was assessed formally. Genome-wide significant SNPs were extracted, and unique variants between studies were grouped by ancestry. Among these, dbSNP-indexed variants were grouped into genomic cluster windows and evaluated for LD structure. Results: Fifteen GWAS were included. Across these studies, 349 genome-wide significant SNP associations were identified, corresponding to 294 unique variants, of which 283 were indexed in dbSNP and retained for genomic and LD analyses. Variant discovery was dominated by large-scale European-ancestry studies, although African, Middle Eastern, East Asian, Hispanic/Latino, South Asian, and trans-ethnic studies also contributed signals. Some evidence of ancestry-specific variation was apparent, yet not conclusive due to lower study power in non-European cohorts. Variant aggregation was strongest at biologically relevant vitamin D loci, including GC, CYP2R1, DHCR7/NADSYN1, and FLG. Fifteen variants were replicated in at least two independent cohorts. LD-based clustering identified several high LD groups comprising variants identified across studies, with the strongest LD appearing between variants within established vitamin D-related loci, particularly GC, CYP2R1, DHCR7/NADSYN1, and FLG. Conclusions: Circulating 25OHD appears to be influenced by shared core loci involved in vitamin D metabolism, across ancestries. Although some evidence of ancestry-specific variation was identified, findings should be interpreted with caution, in light of the predominance of European-ancestry GWAS and scarcity of sufficiently powered GWAS for other ancestral populations. Larger GWAS in non-European populations are essential for improving ancestry-specific variant discovery and interpretation. Full article

The Arabidopsis thaliana Heat Shock Protein-Related (AtHSPR) gene participates in plant growth and abiotic stress tolerance, while its role in biotic stress resistance remains unclear. Here, we report that the athspr mutant is sensitive to Pseudomonas syringae pv. tomato (Pst) DC3000, whereas over-expression of AtHSPR enhances the defense of Arabidopsis against the pathogen. AtHSPR expression was induced by treatment with Pst DC3000, flg22, or salicylic acid (SA). Transcriptome analysis showed that mutation of AtHSPR changed the expression patterns of genes associated with defense response, oxidation–reduction, and SA responses, as well as transcription factors. The biochemical evidence revealed that AtHSPR interacted with Thimet Oligopeptidase 1 (TOP1), which modulated the SA-mediated immune response. Co-expression of AtHSPR and TOP1 showed that the TOP1 protein, normally located in the chloroplasts, gathered around the nucleus in response to a pathogen. After pathogen treatment, dynamic tubular projections (stromules) were present, extending from the chloroplasts toward the nucleus, and TOP1 was observed in the nucleus, together with AtHSPR. The top1athspr double mutant had lower SA levels and was more sensitive to pathogens than the top1 and athspr single mutants. Taken together, our results demonstrated that the interaction between AtHSPR and TOP1 plays a positive role in SA-mediated plant resistance against Pst DC3000. Full article

Type XVII collagen (COL17) is crucial for skin integrity but difficult to produce. To achieve high-level secretory expression, a human COL17 segment was designed and cloned into the pPIC9K vector with six different α-mating factor signal peptides and integrated into Komagataella phaffii GS115. Fed-batch fermentation in a 5 L bioreactor yielded 4.7 g/L of recombinant COL17. Functional assays showed that it promoted the proliferation of human skin fibroblast (HFF) and immortalized keratinocytes (HaCaT), upregulated COL1A1, COL3A1, and TIMP1 in HFF cells, and enhanced skin barrier-related genes (KRT1, KRT5, KRT10, KRT14, IVL, LOR, FLG) in HaCaT cells. In a UVB-induced photoaging model, COL17 reduced reactive oxygen species (ROS) and matrix metalloproteinase 3 (MMP3) activity. This recombinant collagen exhibits photoprotective, regenerative, and barrier-enhancing activities, offering potential for skincare and tissue engineering. Full article

Food allergy (FA) and eosinophilic esophagitis (EoE) represent two of the most rapidly increasing allergen-driven conditions in pediatric medicine. Both diseases share key immunological features, including Th2 polarization and epithelial barrier dysfunction. Over the past two decades, compelling epidemiological and mechanistic evidence has established EoE as a late-manifesting component of the allergic march—the well-recognized sequential progression of atopic disease in childhood, which typically begins with atopic dermatitis, followed by IgE-mediated food allergy, allergic rhinitis, and asthma. Children with IgE-mediated food allergy carry a substantially elevated risk of developing EoE, and shared genetic susceptibility loci—including CAPN14, TSLP, and filaggrin (FLG)—underscore common pathogenic pathways. We conducted a narrative review of the literature by systematically searching PubMed/MEDLINE, EMBASE, and the Cochrane Library using the terms “eosinophilic esophagitis,” “food allergy,” “atopic march,” “IgE-mediated allergy,” and “pediatric” in combination; articles published from 2000 to March 2026 were considered, with priority given to systematic reviews, meta-analyses, randomized controlled trials, and guideline documents. This narrative review comprehensively examines the epidemiology, pathomechanisms, clinical presentation, diagnostic approach, and therapeutic landscape for pediatric FA and EoE, with particular emphasis on their immunological intersections and the evolving evidence positioning EoE within atopic disease trajectories. We highlight approval of dupilumab for children as young as 1 year with EoE—representing a paradigm shift toward biologic therapy for atopic multimorbidity—and discuss the pipeline of emerging agents including cendakimab, lirentelimab, and anti-IL-5 strategies. Identification of shared pathogenic mechanisms offers promising avenues for unified prevention, early diagnosis, and precision therapeutic approaches for children with multiple atopic diseases. Full article

The development of high-performance cathode materials represents a crucial strategy for enhancing the overall electrochemical performance of aqueous alkaline zinc batteries. The rational design of electrode microstructure and chemical composition can synergistically boost the electrochemical reaction activity, ion/electron transport kinetics, and structural stability. In this work, a composite cathode material, FLG@Ni_xS₆/Co₄S₃/Ni-Co(OH)₂, was successfully synthesized via an electrochemical codeposition method. The engineered architecture offers abundant electrochemically active sites, well-defined ion diffusion pathways, and continuous electron conduction networks. Moreover, the strong interaction among the constituent phases effectively regulates and accelerates the redox reaction kinetics. When integrated into an aqueous alkaline zinc battery, the device attains a high specific capacity of 385 mAh g⁻¹ at 2 A g⁻¹, excellent rate capability (287 mAh g⁻¹ at 80 A g⁻¹), a gravimetric energy density of 590 Wh kg⁻¹, a power density of 128.57 kW kg⁻¹, and remarkable cycling stability, with 100% capacity retention maintained after 20,000 cycles. Overall, this study proposes a scalable and rational composite strategy for designing high-performance electrode materials for next-generation electrochemical energy storage systems. Full article

Lectin receptor-like kinases (LecRLKs) are a large subfamily of receptor-like kinases (RLKs), and their N-terminal lectin domain is predicted to reversibly bind to carbohydrates. Within this family, G-type LecRLKs represent a distinct subclass defined by an extracellular S-locus glycoprotein (SLG) domain, which was originally identified for its role in governing self-incompatibility in Brassica species. Emerging evidence suggests that G-type LecRLKs are involved in plant immunity; however, only a small fraction have been functionally characterized, leaving the roles of most family members largely unknown. In this study, we identified RK3 (Receptor Kinase 3) as the most strongly induced gene within the G-type LecRLK clade VI upon infection with Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Through both gain- and loss-of-function analyses, we demonstrated that RK3 positively regulates flg22-induced immune signaling events, including oxidative burst and mitogen-activated protein kinase (MAPK) activation, as well as downstream responses such as defense gene expression and ethylene production. Remarkably, the immune-enhancing activity of RK3 does not require its kinase domain. Critically, both full-length RK3 and a kinase-deleted variant (RK3-ΔK) constitutively interact with FLS2 (Flagellin-Sensing 2) and BAK1 (BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1). This provides direct evidence that RK3 functions primarily as a co-regulatory component within the PRR complex, independent of its kinase activity. Moreover, ectopic expression of RK3 in tomato enhanced resistance to Pst DC3000, highlighting its potential utility in engineering disease resistance in crops. Thus, RK3 reveals a non-canonical, kinase-independent mechanism by which a G-type LecRLK potentiates plant immunity, expanding our understanding of RLK signaling complexity. Full article

Inherited ichthyoses are clinically and genetically heterogeneous disorders of cornification caused by disruption of epidermal barrier genes involved in keratinization and lipid homeostasis. Pathogenic variants in more than 50 genes have been implicated in nonsyndromic ichthyosis vulgaris (IV) and autosomal recessive congenital ichthyosis (ARCI). Here, we investigated the genetic basis of ichthyosis in four consanguineous Pakistani families presenting with IV or ARCI phenotypes. Exome sequencing followed by segregation analysis identified pathogenic variants in four established ichthyosis-associated genes: CYP4F22, FLG, ALOX12B, and NIPAL4. Identified variants include one novel nonsense allele of CYP4F22 (c.296G>A; p.Trp99*) and three known variants previously not reported in the Pakistani population. These known variants include a nonsense change in FLG, a frameshift allele of ALOX12B, and a missense variant in NIPAL4. Standardized phenotypic annotation using Human Phenotype Ontology terms revealed overlapping but variable clinical features across families, consistent with known genotype–phenotype heterogeneity in inherited ichthyosis. In silico protein modeling using AlphaFold2 and Ramachandran plot analysis predicted structural perturbations associated with the identified variants, supporting their pathogenic relevance. Publicly available scRNAseq datasets revealed greater heterogeneity of keratinocyte-associated expression patterns of these ichthyosis-associated genes in aging samples. Collectively, our findings expand the allelic and phenotypic spectrum of inherited ichthyosis in the Pakistani population and highlight the utility of comprehensive genetic analysis in consanguineous families for accurate molecular diagnosis, genetic counseling, and disease epidemiology. Full article

Background/Objectives: The wall-associated kinases (WAKs) and WAK-like proteins (WAKLs) comprise a unique receptor-like kinase subfamily in plants, which have been shown to regulate plant development and defense responses by sensing cell wall-derived components, such as pectin or pectin fragments. In this study, we aimed to characterize the function of WAKL10 in flg22-triggered immunity in Arabidopsis. Methods: Through functional analyses of WAKL genes in Arabidopsis, we identified WAKL10 as the most pronouncedly induced WAKL member in response to flg22 treatment. Gain- and loss-of-function genetic analyses were performed to assess its role in flg22-triggered immune responses, including mitogen-activated protein kinase (MAPK) activation, reactive oxygen species (ROS) burst, and defense gene induction. Transgenic Arabidopsis plants expressing a kinase domain-deleted mutant (WAKL10-ΔK) were generated. Co-immunoprecipitation assays were conducted to examine interactions with FLAGELLIN-SENSITIVE 2 (FLS2) and BRI1-ASSOCIATED RECEPTOR KINASE 1 (BAK1). Heterologous overexpression of WAKL10 in tomato was also tested for bacterial disease resistance. Results: WAKL10 positively regulates flg22-triggered immune responses. Interestingly, WAKL10-ΔK retains the capacity to potentiate these responses. Co-immunoprecipitation assays showed that both wild-type WAKL10 and WAKL10-ΔK constitutively associate with FLS2 and BAK1. Overexpression of WAKL10 in tomato confers enhanced bacterial disease resistance. Conclusions: The extracellular domain of WAKL10 promotes FLS2-BAK1 complex formation, thereby contributing to flg22 signaling. This study reveals a new function of WAKLs, distinguished from their proposed role in sensing cell wall components. The functional conservation of WAKL10 suggests its potential application in engineering disease resistance in crop plants. Full article

Papain-like cysteine proteases (PLCPs) are central immune hubs frequently targeted by pathogen effectors. Sugarcane smut, caused by Sporisorium scitamineum, threatens global sugarcane yield, yet effector manipulation of host PLCPs remains unclear. Genome-wide analysis of Saccharum spontaneum AP85-441 identified 61 PLCP-encoding genes, which were classified into nine conserved subfamilies. Among these, ScRD21A, a member of the RD21 subfamily, was prioritized for functional characterization. Two Pit2 homologs, SsPit2A and SsPit2B, were identified from S. scitamineum. Yeast two-hybrid, BiFC and pull-down assays demonstrated that both effectors interact with ScRD21A, and that this interaction depends on a conserved LXRR motif within their PID14-like region. In total protein extracts from Nicotiana benthamiana, co-expression of SsPit2A or SsPit2B reduced ScRD21A-associated cysteine protease activity. Transient expression of ScRD21A enhanced flg22-induced ROS production, attenuated Pst DC3000-induced hypersensitive response-associated necrosis, and increased resistance to Botrytis cinerea. Together, these results support a conserved PLCP-targeting strategy in smut fungi and identify the ScRD21A–SsPit2A/B module as a tractable framework for studying effector–protease interactions relevant to sugarcane smut. Full article

In this study, we report the draft genome sequence of Pseudomonas aeruginosa strain ASK-80, a multidrug-resistant bacterium isolated from municipal wastewater in Baddi, district Solan, Himachal Pradesh, India. The whole genome was sequenced through Illumina MiSeq sequencing (150 bp paired-end). The size of the assembled genome was 6,261,345 bp, and the genome annotation revealed 5834 genes, including 5778 CDSs, 5748 protein-coding genes, 56 RNA genes and 30 pseudo genes. Genomic characterization revealed the occurrence of multiple antibiotic resistance genes (bla_OXA-396, bla_OXA-486, bla_OXA-494, bla_PAO, bla_PDC-8, aph(3′)-IIb, catB7, fosA and others), virulence genes (algB, chpA, clpV1, exsA, flgA, pilB, pvcA, toxA, tse1, and waaA), insertion sequences, transposable elements and phage sequences. This genome data may serve as a valuable resource for comparative genomics of P. aeruginosa and research on the antibiotic resistance surveillance of wastewater. Full article

Acidithiobacillus caldus is perpetually exposed to multiple extreme environmental stresses. CsrA, functioning as a post-transcriptional regulator of physiological metabolism, acts as a differential modulator, facilitating more economical and efficient adaptation to extreme environments. The csrA expression recombinant strain was constructed in A. caldus MTH-04 by conjugative transfer technology pJD215. Physiological characterization revealed enhanced acid tolerance, significantly elongated flagella, elevated extracellular secretion, and altered biofilm composition. Notably, intracellular concentrations of free glutamate and aspartate increased to 24.18 mg/L and 16.07 mg/L, respectively. The secondary structure of CsrA protein was determined in vitro through circular dichroism spectroscopy and size-exclusion chromatography. Electrophoretic Mobility Shift Assay (EMSA) successfully demonstrated in vitro binding activity of CsrA to the rpoS leader mRNA. CsrA suppresses rpoS mRNA translation by competing with ribosomes for binding sites, thereby negatively regulating rpoS expression. Critical binding sites were further validated through site-directed mutagenesis. Through EMSA, RT-qPCR and the translation reporter system, it was also found that CsrA has a dual regulatory function for nearby flagella- and motility-related gene clusters (flgC, 07035, motD, 15040), which also implies the global regulatory role of CsrA. In summary, a potential overall post-transcriptional regulatory mechanism based on CsrA and rpoS by extremophile A. caldus was proposed. Finally, the efficiency of bioleaching application by csrA overexpression strain was improved by 20.81%. Full article