Search Results (714)

Background: Ectopic fat deposition has been demonstrated to play a critical role in the onset and progression of renal dysfunction. However, research on renal parenchymal fat deposition and its association with renal dysfunction in type 2 diabetes mellitus (T2DM) remains limited, particularly regarding its association with early kidney injury. The present study aimed to further investigate the relationship between renal fat fraction (FF) and biomarkers of kidney injury, thereby providing new evidence for the potential link between intrarenal fat accumulation and early renal impairment in T2DM. Methods: This cross-sectional study enrolled 60 patients with T2DM. Renal FF was quantitatively assessed using magnetic resonance imaging (MRI). Clinical characteristics, body composition parameters, and biochemical indices were collected. Levels of kidney injury biomarkers, including tumor necrosis factor receptors 1 (TNF-R1), tumor necrosis factor receptors 2 (TNF-R2), chitinase-3-like protein 1 (YKL-40), and kidney injury molecule-1 (KIM-1), were measured using enzyme-linked immunosorbent assay (ELISA). To evaluate the correlations between fat distribution and inflammatory biomarkers, Pearson correlation analysis was performed. Furthermore, linear regression analysis was conducted to explore the associations between renal FF and kidney injury biomarkers with adjustments for potential confounders such as smoking status, diabetes duration, and visceral fat. Lasso regression was used to screen variables. Results: The results demonstrated that renal FF was significantly positively correlated with serum YKL-40 (r = 0.3, p = 0.021), TNF-R1 (r = 0.246, p = 0.042), and urinary KIM-1 (r = 0.396, p = 0.004), indicating a close association between renal fat accumulation and early kidney injury biomarkers. In regression analyses adjusted for age, sex, and duration of diabetes, the associations between renal FF and these biomarkers remained significant. After further adjustment for potential confounders, including smoking history, alcohol consumption, hypertension, renin-angiotensin-aldosterone system (RAAS) inhibitors, sodium-dependent glucose transporters 2 (SGLT2) inhibitors, glucagon-Like Peptide-1 (GLP-1) receptor agonists, and lipid-lowering drugs, renal FF remained significantly associated with TNF-R1 (β = 0.327, p = 0.015), KIM-1 (β = 0.352, p = 0.021), and YKL-40 (β = 0.275, p = 0.025). Moreover, even after additional adjustment for visceral fat, the associations of renal FF with TNF-R1 and KIM-1 persisted. After using the Benjamini–Hochberg procedure for false discovery rate, the relationship between renal FF and KIM-1 had a significant difference. Variables of age and gender were excluded to build the parsimonious modeling using Lasso regression. It suggested that renal fat accumulation may contribute to kidney injury independently of visceral adiposity. Conclusions: The study systematically demonstrates a significant association between renal FF and early biomarkers of kidney injury in T2DM, which may suggest the potential role of renal fat accumulation in the pathogenesis of diabetic nephropathy. These findings provide clinical data support for the development of a fat-targeted intervention study. Future research should further elucidate the long-term mechanistic role of renal FF in diabetic nephropathy, as well as its potential value in early diagnosis and therapeutic applications. Full article

To identify chitinase genes from the genome of the mycoparasitic Cladosporium sp. strain SYC23, bioinformatical analyses and real-time quantitative PCR (RT-qPCR) were employed to screen mycoparasitism-associated genes at 12, 24, 48, and 72 h post-induction with Aecidium pourthiaea rust spores. A total of eight chitinase genes were identified from SYC23 via bioinformatics analysis and designated CaChi34, CaChi40, CaChi45, CaChi67, CaChi82, CaChi92, CaChi93, and CaChi286 based on sequence and phylogenetic analyses. Analysis of the chitinase protein sequence characteristics revealed molecular weights ranging from 33.86 to 286.03 kDa and theoretical isoelectric points from 4.48 to 7.7. All CaChi genes contained the conserved GH18 domain, and promoter analysis showed that each harbored MYB-binding sites and pathogen-responsive elements. Mycoparasitism-related sequence clustering analysis indicated that the chitinase sequences of SYC23 shared the closest phylogenetic relationship with those from Trichoderma sp. RT-qPCR results following rust spore induction showed that five CaChi genes reached their highest expression levels at 24 h post-induction, CaChi45 was most highly expressed at 72 h post-induction, CaChi93 was continuously upregulated, and CaChi82 was continuously downregulated throughout the induction period. His-tagged recombinant CaChi93 protein was purified from E. coli and characterized. The results demonstrate that the enzymatic activity of CaChi93 was 0.929 U/mg, with optimal reaction conditions at 65 °C and pH 7. Treatment of A. pourthiaea rust spores with the recombinant CaChi93 chitinase confirmed that CaChi93 could effectively dissolve rust spore walls. In conclusion, this study confirms that the mycoparasitic Cladosporium sp. strain SYC23 can secrete chitinase to degrade the rust spore wall and induce spore death, thereby providing novel gene resources and a theoretical basis for the biological control of A. pourthiaea. Full article

Saliva is an important biological matrix that allows the investigation of various welfare parameters; in ruminants, it is abundant and can be easily collected without requiring professional veterinary intervention. The aim of the investigation was to provide additional information on both digestive and antioxidant enzymes of sheep farmed in good welfare conditions, assessed with the Animal welfare indicators (AWIN) protocol. Small surgical forceps holding a Salivette^® device (Sarstedt, Nümbrecht, Germany) were gently inserted into the sheep’s mouths without any force. The sheep chewed the swabs independently for a few seconds, allowing saliva collection. Seventeen enzymes from the saliva of 15 sheep were analyzed: antioxidant enzymes (catalase, glutathione S transferase), proteolytic enzymes (trypsin, chymotrypsin, N-aminopeptidases, carboxypeptidase A and B), carbohydrases (glucose oxidase, amylase, cellulase, lignin peroxidase, chitinase and α-glycosidase), and esterases (alkaline and acidic phosphatases, lipase and esterase). Esterase activity showed the highest value (12.95 ± 1.25 U/mg of proteins), whereas lignin peroxidase activity showed the lowest (2.23 ± 0.37 µU/mg of proteins). The activity of all enzymes was observed except for glutathione S transferase and α-glycosidase. Among the enzymes, lipase activity has already been identified as a biomarker of stress in sheep saliva. This investigation may represent a basis for further investigations into the diet and adaptive responses of sheep to different environmental conditions. Furthermore, samples collected using the Salivette^® device can be easily obtained without requiring specialized staff and without causing any stress to the animals. Further investigations into the origin of individual enzymes using a proteomic approach are desirable. Full article

The growing emphasis on sustainability, regional distinctiveness, and spontaneous fermentation in winemaking necessitates a more comprehensive understanding of local yeast populations and their functional mechanisms. In total, 115 indigenous yeast strains were examined for their enzymatic activities of potential vitivinicultural significance. The yeasts were screened for chitinase activity (biocontrol potential), glycosidase activity (terpene release), β-lyases (thiol release), and sulfite reductases (off-flavor formation), followed by quantitative analysis of the selected subsets. Yeasts were further evaluated for inhibition of fungal mycelial growth, VOC-mediated inhibition, and tolerance to commonly applied fungicides. Pre-field selection was refined using the niche overlap index and grapevine leaf disc assay. The results confirmed chitinolytic activity in four species; all strains exhibited hydrolase activities, with H. uvarum 116 displaying the highest cell-associated activity (6.32 U/g), while T. delbrueckii Sut94 showed the highest extracellular activity (1.36 U/g). β-glucosidase and β-lyase activities were widespread, whereas hydrogen sulfide production was infrequent. P. guilliermondii ZIM 624 showed the most comprehensive overall enzymatic profile, together with strong inhibition patterns. A field trial on Pinot cultivars (V. vinifera L.) further evaluated P. guilliermondii ZIM 624 within an integrated disease management approach, with responses being more pronounced in ‘Pinot noir’ than in ‘Pinot gris’. Full article

Acidic chitinase (Chia) degrades chitin, a structural polysaccharide in insect exoskeletons, and plays important roles in omnivorous and insectivorous mammals and birds. In birds, gene duplications have generated multiple Chia paralogs with functional divergence, but the molecular basis for this diversification remains unclear. Here, we characterized three chicken Chia paralogs (Chia1–3) and identified distinct pH-dependent enzymatic profiles. Chia1 is enzymatically inactive but was captured by chitin-affinity resin despite lacking a canonical chitin-binding domain, suggesting residual substrate interaction through the catalytic domain or a non-catalytic role. Chia2 exhibits maximal activity at pH 2.0, whereas Chia3 peaks at pH 5.0 and displays broader activity. Exon swapping and site-directed mutagenesis identified residues 104 (Ala in Chia2, Asp in Chia3) and 269 (His vs. Asn) as key contributors to pH-dependent activity differences. Reciprocal substitutions shifted pH profiles accordingly. Structural modeling and computational pKa predictions suggested that D213 and residue 269 may function as a pKa-regulating module influencing catalytic ionization. Comparative sequence analysis revealed lineage-specific conservation of these residues, consistent with adaptive divergence. Our findings show that limited amino acid substitutions can markedly modify pH-dependent enzymatic activity, providing mechanistic insight into how local residue variation contributes to the functional diversification of duplicated genes. Full article

Postharvest Lentinula edodes (shiitake mushroom) undergoes rapid textural deterioration, which is primarily driven by complex cell wall remodeling. This study investigates the physiological and transcriptomic changes in L. edodes during storage at 4 °C for 8 days. Results showed that cellulose content significantly decreased, while chitin and β-glucan levels exhibited anomalous increases, accompanied by a surge in the activities of cellulase, chitinase, and β-1,3-glucanase. Concurrently, intensifying membrane lipid peroxidation and an imbalance in reactive oxygen species (ROS) homeostasis were observed. Transcriptomic analysis identified 2204 and 1808 differentially expressed genes (DEGs) at the middle (4 d) and late (8 d) storage stages, respectively. Partial Least Squares Regression (PLSR) identified a core module of nine key regulatory genes (VIP > 1.0), including β-glucanase, laccase, and catalase, which significantly contributed to the physiological shifts. The results suggest that an upstream ROS imbalance may contribute to the dysregulation of midstream laccases, potentially reducing the oxidative cross-linking of phenolic components and loosening the cell wall matrix. These alterations may increase the accessibility of structural polysaccharides to downstream cell wall-degrading enzymes, which could contribute to structural collapse, although functional validation is required to establish causality. These findings provide a gene-level framework for understanding postharvest edible fungi physiology. Full article

The molting of crustaceans is accompanied by exoskeleton reconstruction. To reveal the molecular regulation mechanism of exoskeleton remodeling, the transcriptomic profiles of the exoskeleton across the entire molting process in the red swamp crayfish Procambarus clarkii were investigated by RNA sequencing, yielding a total of 7671 differentially expressed genes (DEGs) across five different molting stages. Notably, the key DEGs were those related to cuticular exoskeleton synthesis (cuticular proteins), degradation (chitinase 2, chitinase 10) and hardening (chitin deacetylase 1), and their expression abundance varied by 10-fold or greater across the molting cycle. Analysis of Gene Ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) showed that significantly enriched pathways included the structural constituents of the cuticle, structural molecule activity, chitin binding of chitin metabolism, and hormone biosynthesis. The expression profiles of nine selected molting-related DEGs were further validated via real-time RT-PCR assays. The acquired unique temporal expression patterns involved in exoskeleton remodeling provide a preliminary insight into the regulation of gene expression during the molting cycle in the red swamp crayfish. Full article

Background: Monitoring inflammatory bowel disease (IBD) during pregnancy is challenging due tolimited use of invasive tools. While fecal calprotectin is considered reliable, its use is limited by patient adherence and availability. Blood-based markers, such as serum chitinase-3-like-1 (CHI3L1), offer a promising alternative. Aim: To evaluate whether serum CHI3L1 reflects disease activity in pregnant IBD patients, compared to standard markers and clinical questionnaires. Methods: Pregnant IBD patients were recruited from a multidisciplinary clinic. Blood samples were collected to assess inflammatory markers. Stool samples were used to measure calprotectin levels. Each visit was classified as a distinct sample for analysis. Correlations between CHI3L1 and disease activity markers were examined. Results: A total of 124 samples from 80 pregnant IBD patients were analyzed: 90 from Crohn’s disease (CD) patients and 34 from ulcerative colitis (UC) patients. CHI3L1 levels showed a significant positive correlation with fecal calprotectin (r_p = 0.366, p = 0.008), ESR (r_p = 0.358, p = 0.001), CRP (r_p = 0.478, p < 0.001) and standardized clinical scoring questionnaires. Elevated CHI3L1 (>56.6 ng/mL) is a risk factor for active disease (OR 8.78, 95% CI 1.54–49.83, p = 0.014). Conclusions: Serum CHI3L1 is positively associated with established markers of inflammation and may serve as a useful non-invasive biomarker for monitoring IBD activity during pregnancy, a medical condition in which invasive procedures are not recommended. Based on CHI3L1 levels, personalized treatment for pregnant IBD patients can be tailored. However, further validation is recommended. Full article

Background: Sirtuin 3 (SIRT3), a mitochondrial NAD⁺-dependent deacetylase, plays a central role in regulating mitochondrial metabolism, oxidative stress, and cell survival. Although SIRT3 has been implicated in angiogenesis, apoptosis, and inflammation, its global proteomic impact on the brain remains unclear. This study aimed to systematically characterize alterations in angiogenesis-, apoptosis-, chemokine-, and cytokine-related proteins in the brains of SIRT3 knockout (SIRT3 KO aka SIRT3⁻/⁻) mice compared with wild-type (WT) controls. Methods: Adult male C57BL/6 WT and SIRT3 KO mice were analyzed using proteome profiler antibody microarrays covering 53 angiogenesis factors, 21 apoptosis markers, 28 chemokines, and 111 cytokines. Protein expression changes were quantified by chemiluminescence imaging and densitometric analysis. Results: The results showed a distinct suppression of angiogenic proteins (amphiregulin, angiogenin, DPPIV, GM-CSF, IGFBP-2, IGFBP-3, IL-1β, PDGF-AA, PDGF-BB, proliferin, serpin F1, thrombospeondin-2, TIMP-4, and VEGF-B), activation of both pro-apoptotic (BAD, cytochrome c, Smac/DIABLO, HIF-1α, Fas, TNF R1, and TRAILR2) and anti-apoptotic, stress-related proteins (Bcl-x, catalase, HO/HMOX2, HSP27, HSP70, and MCL1) in the SIRT3 KO animals compared with the WT controls. Notably, SIRT3 deficiency was associated with increased expression of inflammatory mediators linked to glial activation and neurodegeneration (BLC/CCL13, LIX/CXCL5, MIG/CXCL9, chitinase 3-like 1, CCL22/MDC, IL-6, myeloperoxidase, osteopontin, RBP4, Reg3G, and TNF-α), alongside disturbed proteins involved in immune surveillance and vascular remodeling (6Ckine/CCL21, chemerin, DF, EGF, fractalkine/CX3CL1, HGF, IGFBP-6, IL-16, and I-TAC). Conclusions: Collectively, these findings demonstrate that SIRT3 is a key regulator of mitochondrial-dependent vascular, apoptotic, and neuroimmune pathways in the brain, and that its loss creates a molecular environment consistent with heightened vulnerability to neurodegenerative processes. Full article

The cactus pear (Opuntia spp.) is a crop of major economic and ecological importance in arid and semi-arid regions. However, with its domestication and intensification, symptoms of fungal diseases have begun to emerge in different cultivation areas. This study was conducted to identify the pathogenic fungi associated with symptoms observed on cladodes in different regions of Morocco and to evaluate the effectiveness of bacterial and fungal antagonists. The study enabled the isolation and identification of several fungal agents from symptomatic cladodes, namely Alternaria alternata, Alternaria tenuissima, Colletotrichum gloeosporioides, and Aspergillus tubingensis. Among these pathogens, A. alternata proved to be the most aggressive and was therefore selected for in vitro and in vivo antagonism assays. Twelve bacterial isolates belonging to the genera Bacillus and Pseudomonas, as well as one isolate of Trichoderma harzianum, were evaluated for their antifungal activity. All antagonists showed significant inhibitory effects against A. alternata in vitro preliminary assay. However, the bacterial isolates B. siamensis, B. halotolerans, and P. peli, as well as T. harzianum, exhibited the highest efficacy. This efficacy was confirmed through direct confrontation tests in vivo on one-year-old cladodes for the three bacterial isolates. In contrast, T. harzianum showed significant pathogenic potential on cladodes of O. ficus-indica and O. megacantha. Investigation of the mechanisms of action of the three most effective bacterial isolates revealed their ability to produce antifungal volatile organic compounds. Enzymatic analyses showed differential production of amylase, chitinase, cellulase and protease among the three isolates, while genes associated with the biosynthesis of antifungal lipopeptides were detected only in P. peli. Full article

This study investigated the expression and subcellular localization of chitinase-3-like protein 1 (CHI3L1) in neutrophils and plasma from untreated and dimethyl fumarate (DMF)-treated multiple sclerosis (MS) patients, and healthy controls. Intracellular CHI3L1 expression was assessed in CD66b+ neutrophils and CD16+ cells using flow cytometry. Subcellular localization was analyzed by confocal microscopy using markers for various neutrophil granules, while ELISA measured plasma CHI3L1 and lactoferrin levels. We found that both intracellular CHI3L1 levels (expressed as mean fluorescence intensity, MFI) and the proportion of CD66b+ cells were significantly increased in MS patients compared to healthy controls. CHI3L1 was found to colocalize with CD66b+ specific granules. While plasma CHI3L1 levels in untreated MS patients remained comparable to those of healthy controls, a significant increase in both intracellular and plasma CHI3L1 was observed in DMF-treated MS patients. The lack of correlation between plasma lactoferrin and CHI3L1 might suggest selective release mechanisms or differential synthesis of these proteins, despite their common storage in specific granules. These findings highlight the role of neutrophils as a peripheral source of CHI3L1 and suggest a complex association between neutrophil-derived CHI3L1 and the differences observed in DMF-treated MS patients. Full article

PET biodegradation remains limited due to its intrinsic properties—high crystallinity, hydrophobicity, and strong chemical stability. These characteristics lead to extremely slow degradation rates and contribute to PET’s persistence in the environment. Understanding how microorganisms respond at the molecular level when exposed to such a recalcitrant polymer is therefore essential. Living organisms express genes in response to their needs during development. When microbes are under critical conditions, such as when contaminants are present, they express genes encoding specific enzymes that attack the pollutant. In this study, a fungus isolated from the infected fruit of the plant Randia monantha was identified as Aspergillus terreus. It was tested for polyethylene terephthalate (PET) degradation, and the fungus Aspergillus nidulans was evaluated due to its previously reported recombinant cutinases for PET degradation. A microplastic polyethylene terephthalate (PET-MP) particle size of <355 μm for degradation was established, and a PET weight loss of 1.62% for A. nidulans and 1.01% for A. terreus was found. Additionally, the degradation of PET was confirmed by FTIR and SEM. This study also compares the transcriptomic profiles of Aspergillus nidulans and Aspergillus terreus during cultivation with PET-MP residues, which serve as a replacement for the carbon source. We present the first evidence of chitinase overexpression during direct exposure of PET to Aspergillus fungi. Interestingly, chitinase expression was detected in the crude extracts of A. nidulans and A. terreus during culture in the presence of PET residues, which replaced the carbon source. The chitinase produced by each fungus has a similar molecular weight of approximately 44 kDa. Chitinase activity was monitored over a 14-day cultivation period; from day 2, chitinase activity was detected in both cultures and continued to increase until day 14, when the highest values reported in this work were 24.88 ± 4.17 U mg⁻¹ and 10.41 ± 0.47 U mg⁻¹ for A. nidulans and A. terreus, respectively. Finally, we proposed a pathway for PET degradation by Aspergillus fungi that involves mycelial adherence and the secretion of hydrophobins, followed by the production of intermediates and monomers via esterase hydrolysis, and ultimately, the entry of monomers to the ethylene glycol (EG) and terephthalic acid (TPA) pathways, further suggesting these Aspergillus as candidates to produce valuable compounds under these conditions, such as muconic acid, gallic acid, and vanillic acid. Full article

Chordomas are rare, highly morbid tumors arising from notochordal progenitor cells along the spinal axis, associated with severe neurological complications and high recurrence rates. Their resistance to conventional therapies and limited options beyond surgical resection and high-dose radiation underscore the urgent need for novel therapeutic targets. Publicly available preliminary RNA sequencing data from the Chordoma Foundation identified chitinase-3-like 1 (CHI3L1), a secreted glycoprotein implicated in immune checkpoint regulation and epithelial–mesenchymal transition (EMT), as a promising candidate for chordoma immunotherapy. Yet, the comprehensive function of CHI3L1 in chordoma immune response remains unclear. To evaluate its presence in chordoma, we employed RNA-based analyses alongside enzyme-linked immunosorbent assays (ELISA) on commercially available chordoma cell lines (JHC7, U-CH12, U-CH1, U-CH1-N) and human chordoma tumor specimens. Our results demonstrate elevated CHI3L1 expression in chordoma cells relative to notochordal precursors, with comparative analyses revealing higher CHI3L1 expression in the primary tumor relative to recurrent samples. These findings suggest the potential role of CHI3L1 in chordoma tumorigenesis, emphasizing its relevance as a biomarker and therapeutic target for primary tumors. Future studies are necessary to elucidate the mechanistic role of CHI3L1 in chordoma immune evasion and to explore targeted interventions that may improve patient outcomes in this aggressive cancer. Full article

Among the most important species of Ficus L. genus are F. deltoidea, F. exasperata, F. sycomorus, F. religiosa, F. microcarpa, F. hirta Vahl., F. benghalensis, F. racemosa, F. elastica, and F. carica. The genus has more than 30 traditional ethnomedicinal uses, attributed to the combination of different bioactive compounds, including flavonoids, (flavanols, flavones, flavonols, isoflavones, chalcones, anthocyanins), phenolic acids (hydroxycinnamic acids, hydroxybenzoic acids), terpenes (triterpenes, tetraterpenes, diterpenes, sesquiterpenes, monoterpenes), phytosterols, coumarins, hydroxybenzoates, phenylpropanoids, chlorins, pheophytins, megastigmans, chitinases, organic acids, fatty acids, amino acids, alkaloids, and glycosides. With this in mind, the objective of this manuscript was to conduct a scientific search in the main electronic databases (PubMed, SciELO, Latindex, Redalyc, BiologyBrowser, ScienceResearch, ScienceDirect, World Wide Science, Web of Science, Academic Journals, Etnobotany, Scopus, and Google Scholar) to gather information on published research regarding the genoprotective potential of the Ficus L. genus. Unlike most scientific articles, which primarily describe the individual characteristics and properties of each species, this document compiles the largest number of studies (in vitro and in vivo) on Ficus plants described by different authors. Thus, we aim to promote more detailed scientific research and expand studies on the protective capacity of these angiosperm plants to the genetic material. Full article

Tetranychus urticae is a globally important economic pest mite. Isaria cateniannulata can infect the mite through its conidial penetration of the cuticle, ultimately leading to host mortality; however, the immune mechanisms involving enzyme activity systems and chitinase genes of T. urticae during this process remain unclear. In this study, T. urticae infected with I. cateniannulata was used as experimental material to analyze changes in antioxidant and detoxification enzyme activities during infection. In addition, the expression patterns of six chitinase genes were analyzed, and significantly upregulated genes were selected for bioinformatics analysis and functional verification. The results showed that infection with I. cateniannulata enhanced the activity of defense-related enzymes in T. urticae, with Catalase (CAT) and mixed function oxidase (MFO) playing dominant roles. All six chitinase genes were activated, among which TuCHT7 and TuCHT12 were significantly upregulated at 24 h post-infection, and then TuCHT7 gradually declined. Whereas TuCHT12 maintained a sustained and stable induction pattern, TuCHT10 was suppressed, while the other genes exhibited transient expression. Feeding dsTuCHT12 suppressed the expression of this gene within 72 h, with significant suppression observed at 48 h. At this time, the expression levels of TuCHT8, TuCHT9 and TuCHT10 genes were activated, whereas TuCHT7 and TuCHT11 were suppressed. The combined application of dsCHT12 and I. cateniannulata increased the mortality of T. urticae by 3.19-fold and reduced egg production by 52.8%. This study preliminarily revealed the defense response mechanism of T. urticae against I. cateniannulata infection and provides a theoretical basis for eco-friendly pest control based on the combined application of RNAi and arthropodpathogenic fungi. Full article