Search Results (294)

Biostimulants, particularly single amino acids, can increase plant growth and crop quality, gaining significant attention. This study investigates the effects of 10 amino acids via root/foliar application on the growth, quality, taste, and volatile flavor of mini-watermelons and compares the differences between the application methods. Here, we employed electronic noses, electronic tongues, and gas chromatography–ion mobility spectrometry to investigate these effects. Root application excels in fruit growth and pectin accumulation, while foliar application boosts soluble protein and specific nutrients. Specifically, root application (except for Val) significantly increases fruit weight, with Gly being most effective for longitudinal diameter, while most amino acids (except Val/Lys) promote transverse diameter. Pectin content shows bidirectional regulation: root application of Glu/Gly/Lys/Pro/Trp/Val enhances pectin, whereas foliar application inhibits it. For taste indices, most treatments improve soluble solids (except Glu root/Arg-Leu foliar), and Ala/Asp/Glu/Gly reduce titratable acids, optimizing the sugar–acid ratio. Foliar application is more efficient for soluble protein accumulation (Ala/Glu/Gly/Pro/Leu). For nutritional quality, except for Lys, all treatments increase vitamin C and widely promote total phenolics and lycopene, with only minor exceptions, and only Arg foliar application enhances ORAC. Additionally, the results revealed that root-applied lysine and valine greatly raised the levels of hexanal and 2-nonenal, whereas foliar-applied valine significantly increased n-nonanal and (Z)-6-nonenal. Overall, we found that amino acids can considerably improve mini-watermelon production, quality, taste, and antioxidant capacity, providing theoretical and practical references for their widespread use in agriculture. Full article

Background: Prostate cancer is a common malignancy among men worldwide, with various histopathologic features that influence its progression and prognosis. One such feature is perineural invasion (PNI), which has been associated with aggressive disease. In this retrospective study, we analyzed genomic alterations associated with PNI in patients who underwent radical prostatectomy. Methods: A total of 421 prostate cancer patients who underwent radical prostatectomy without neoadjuvant therapy were identified from The Cancer Genome Atlas. PNI was present in 378 patients (89.8%) and absent in 43 (10.2%). Differentially expressed genes were identified, and mRNA expression levels of key genes were analyzed. The prognostic significance of these genes was evaluated using log-rank tests and Cox proportional hazards models to estimate hazard ratios and 95% confidence intervals. Results: Levels of COL9A3, ASPN, ESR1, MUC1, PIP, SFRP4, KRT19, CLDN1, and COMP were significantly higher in the tumor tissues of patients in the PNI group compared to those in the non-PNI group (q < 0.05), and RYR2, MME, and AZGP1 expression levels were significantly higher in the non-PNI group (q < 0.05). A high mRNA expression level of AZGP1 was associated with longer disease-free survival, whereas high mRNA expressions of ASPN, COMP, RYR2, and SFRP4 were associated with shorter disease-free survival. Conclusions: Prostate cancer patients with genomic alterations associated with PNI may face a higher risk of disease progression after prostatectomy, highlighting the need for further prospective studies to validate these findings. Full article

Rice-field eel (Monopterus albus) slices, an important aquatic product in Southeast Asia, are prone to spoilage and deterioration during cold chain storage. In this study, the effects of a composite preservative (ε-polylysine, Vitamin C (Vc), epigallocatechin gallate (EGCG), and phloretin) on the muscle quality (color, texture, water holding capacity (WHC)) of rice-field eel slices during refrigeration storage at 4 °C for up to 7 days was investigated, and the underlying mechanism was elucidated by the integrated microbiome and metabolomics, in addition to Elisa and Low-Field Nuclear Magnetic Resonance (LF-NMR). After 7 days of storage, the WHC, shear force, and a* decreased by 11.39%, 34.37%, and 49.20% in treated samples, and by 19.18%, 38.38%, and 54.87% in control samples, respectively. The addition of the composite preservative significantly increased Hexokinase, Pyruvate kinase, and Creatine kinase, while it decreased the total viable count (TVC), total volatile basic nitrogen (TVB-N), thiobarbituric acid reactive substance (TBARS), and Lactic acid. Preservative treatment maintained the moisture content of the eel slices during storage and prevented bright red oxymyoglobin from transforming into brown metmyoglobin. Microbiota composition (especially Pseudomonas) and metabolic pathways (including amino acid and its metabolites, nucleotide and its metabolite, and organic acid and its derivatives, etc.) were obviously altered by the preservative treatment. Pseudomonas, tryptophan-aspartic acid (Trp-Asp), D-Glucose 6-phosphate, Succinic Acid, Biliverdin 1, 5-Diaminopentane, and Tyramine, etc., are potential biomarkers for the quality changes of eel slices during refrigeration. These findings provide an in-depth understanding of the improvement of the eel slice quality during refrigeration storage by the composite preservative. Full article

Osteosarcoma (OSA) is the most prevalent bone malignancy in people and dogs. Current survival rates show the need for advances in novel therapies to help overcome the growth, survival and metastatic progression of the cancer. Canine models are often used to advance prognostic and treatment opportunities for OSA due to the similarities in the disease between species. This study focusses on the genetic and molecular similarities of OSA between human and canine specimens. Differentially expressed genes (DEGs) were compared and identified in canine and human OSA tumours, revealing 86 common genes, 36 having high and 50 having low expression. Further immunohistochemical analysis of the corresponding proteins of three identified DEGs (ASPN, STK3, BAMBI) allowed for the visualisation of protein expression in canine OSA tissues (n = 19). Overall nuclear and cytoplasmic H-scores were generated, and nuclear and cytoplasmic scores in males and females and in different anatomical locations (axial versus appendicular) were also investigated, presenting unique opportunities to understand the expression in this cancer type. This study contributes to a deeper knowledge of genetic pathways changes and identifies avenues for the diagnosis, prognosis and treatment of OSA in people and dogs, whilst encompassing the One Health concept in medicine. Full article

Xylanases, important enzymes in the food industry, have severely limited use in industrial applications due to insufficient thermal stability. This study focused on improving the thermostability of XynASP, a glycoside hydrolase family 11 (GH11) xylanase from Aspergillus saccharolyticus JOP 1030-1, through modular assembly and rational mutagenesis. By aligning XynASP with nine thermostable GH11 homologs, six variable structural modules (β1, β3, β6, β7, α1, β14) and eight non-conserved residues were identified. Six chimeras (Z1, Z2, Z3, Z4, Z5, Z6) and eight single mutants (S131T, Y133T, A137G, A144T, T147Y, A156R, V198M, and Y204Q) were constructed. Among these, the β3-module-substituted chimera Z2 exhibited a 15.4-fold extended half-life at 45 °C compared to wild-type XynASP. Single-point mutagenesis revealed that V198M showed the highest residual activity after thermal treatment. To further optimize stability, combinatorial mutagenesis was performed: the double mutant A144T/V198M demonstrated a 4.3-fold longer half-life at 50 °C. Combining Z2 with the A144T/V198M mutations yielded the chimeric ZM, which demonstrated a 26.5-fold increase in half-life at 50 °C and a 5.5-fold improvement in catalytic efficiency (197.4 U/mg) compared to wild-type XynASP. Structural analysis and molecular dynamics simulations showed that increased hydrophobic interactions at both the N- and C-termini improved the structural stability of chimeric ZM, while increasing the flexibility of the thumb can offset the negative impact on catalytic activity during thermal stability modification of GH11 xylanase. This study further confirmed that modular assembly is an effective approach for obtaining high-activity, heat-resistant xylanases. This study also notably deepened our understanding of the thermal stability mechanisms of xylanases. Full article

Tetanus, a severe and life-threatening illness caused by Clostridium tetani, produces symptoms such as muscle spasms, muscle stiffness and seizures caused by the production of tetanus neurotoxin (TeNT). TeNT causes spastic paralysis through the inhibition of neurotransmission in spinal inhibitory interneurons. This is achieved, in part, through pH-triggered membrane insertion of the translocation (HCT) domain, which delivers the catalytic light-chain (LC) domain to the cytosol. While the function of HCT is well defined, the mechanism by which it accomplishes this task is largely unknown. Based on the crystal structure of tetanus neurotoxin, we identified potential polar interactions between arginine 711, tryptophan 715 and aspartate 821 that appear to be evolutionarily conserved across the clostridial neurotoxin family. We show that the disruption of the Asp-Arg pair in a beltless HCT variant (bHCT) results in changes in thermal stability without significant alterations to the overall secondary structure. ANS (1-anilino-8-napthalene sulfonate) binding studies, in conjunction with liposome permeabilization assays, demonstrate that mutations at R711 or D821 trigger interactions with the membrane at higher pH values compared to wildtype bHCT. Interestingly, we show that the introduction of the D821N mutation into LH_NT (LC-HCT only), but not the holotoxin, resulted in the increased cleavage of VAMP 2 in cortical neurons relative to the wildtype protein. This suggests that, as observed for botulinum toxin A, the receptor-binding domain is not necessary for LC translocation but rather helps determine the pH threshold of membrane insertion. The mutation of W715 did not result in detectable changes in the activity of either bHCT or the holotoxin, suggesting that it plays only a minor role in stabilizing the structure of the toxin. We conclude that the protonation of D821 at low pH disrupts interactions with R711 and W715, helping to drive the conformational refolding of HCT needed for membrane insertion and the subsequent translocation of the LC. Full article

Background: Antimicrobial resistance (AMR) is a global threat in the public healthcare sector. The emergence of carbapenem-resistant Enterobacterales (CRE) has become a serious public health threat in South Africa. The spread of CRE has led to the use of colistin for treating severe infections. Colistin is a cationic, lipopeptide antibacterial agent that is effective against most Gram-negative bacteria through its disruption of the bacterial cell membrane. This study aims to determine the colistin resistance (MIC) and mobile colistin resistance (mcr-1) gene in clinical isolates from healthcare facilities in Mthatha and its surrounding areas. Methods: Fifty-three CRE isolates were collected from health facilities between January 2019 and June 2021 and stored in skim milk 10% and 5% inositol broth. The carbapenemase confirmatory test involved a RESIST-4 O.K.N.V assay (Coris BioConcept, Gembloux, Belgium), which was conducted following manufacturer protocol. Broth microdilution was performed according to the ISO standard method (20776-1) using A ComAspTM colistin 0.25–16 μg/mL MIC Broth. Conventional polymerase reaction (PCR) was performed for the detection of mcr-1. Results: N = 53 (100%) isolates were used. A total of 53% were defined as Klebsiella pneumoniae, Escherichia coli constituted 8%, Enterobacter cloacae 8%, Serratia marcescens 8%, Serratia fonticola 2%, Enterobacter aerogenes 2%, Klebsiella oxytoca 2%, Citrobacter koseri 2%, and Citrobacter freundii 2%. The specimens were from the following wards: Pediatric and Neonatal 38%, Medical 30%, Gynecology, Labour, and Maternity 11%, OPD and A&E 11%, ENT 4%, and Others—Male TB ward, Trauma, and adult ICU 6%. In total, 13% of the isolates were resistant and 86% were sensitive to colistin. The common CRE genes detected were OXA-48 at 47%, NDM at 13%, VIM at 1%, and a combination of OXA-48 and NDM at 5%. Of the isolates, 66% were positive for the production of carbapenamase. In this study, we found that all N = 53 (100%) isolates did not have the mobile colistin resistance gene (mcr-1). Conclusions: Antimicrobial resistance is associated with the emergence of carbapenemases genes. Increasing resistance to colistin in clinical settings can lead to difficulties in treating CRE infections, which may lead to clinical failure. In our study, 13% of isolates were phenotypically resistant to colistin. Full article

The m.13513G>A (p.Asp393Asn) substitution in the MT-ND5 (Mitochondrially Encoded NADH/Ubiquinone Oxidoreductase Core Subunit 5) gene is a common pathogenic variant associated with primary mitochondrial disorders. It frequently causes Leigh syndrome and mitochondrial encephalomyopathy with lactate acidosis and stroke-like episodes (MELAS). In this study, we present clinical data, heteroplasmy levels in various tissues (blood, urine, and skin fibroblasts), and bioenergetic characteristics from a cohort of 20 unrelated patients carrying the m.13513G>A mutation, classified according to the following phenotypes: Leigh syndrome (n = 12), MELAS (n = 2), and Leber’s hereditary optic neuropathy (LHON, n = 6). We observed a significant correlation between high respiratory ratios and heteroplasmy levels in fibroblast cell lines of the patients. Furthermore, fibroblast cell lines with heteroplasmy levels exceeding 55% exhibited markedly reduced mitochondrial membrane potential. These findings contribute to a better understanding of the clinical and bioenergetic profiles of patients with m.13513G>A-variant-related phenotypes across different heteroplasmy levels, based on data from a single genetic center. Our data suggest that even a slight shift in heteroplasmy can improve cellular function and, consequently, the patients’ phenotype, providing a solid foundation for the development of future gene therapies for mtDNA diseases. Full article

Current weakly supervised salient object detection (SOD) methods for RGB-D images mostly rely on image-level labels and sparse annotations, which makes it difficult to completely contour object boundaries in complex scenes, especially when detecting objects with filamentary structures. To address the aforementioned issues, we propose a novel cross-modal weakly supervised SOD framework. The framework can adequately exploit the advantages of cross-modal weak labels to generate high-quality pseudo-labels, and it can fully couple the multi-scale features of RGB and depth images for precise saliency prediction. The framework mainly consists of a cross-modal pseudo-label generation network (CPGN) and an asymmetric salient-region prediction network (ASPN). Among them, the CPGN is proposed to sufficiently leverage the precise pixel-level guidance provided by point labels and the enhanced semantic supervision provided by text labels to generate high-quality pseudo-labels, which are used to supervise the subsequent training of the ASPN. To better capture the contextual information and geometric features from RGB and depth images, the ASPN, an asymmetrically progressive network, is proposed to gradually extract multi-scale features from RGB and depth images by using the Swin-Transformer and CNN encoders, respectively. This significantly enhances the model’s ability to perceive detailed structures. Additionally, an edge constraint module (ECM) is designed to sharpen the edges of the predicted salient regions. The experimental results demonstrate that the method shows better performance in depicting salient objects, especially the filamentary structures, than other weakly supervised SOD methods. Full article

Dried shrimp is a popular dietary ingredient that is often included in appetizer soups, stir-fry dishes, or other stews to improve the umami taste. The effects of adding dried shrimp on the sensory characteristics and taste components of sheep bone soup were investigated through sensory evaluation and untargeted approaches. The results of the single-factor and orthogonal experiments showed that the flavor qualities of sheep bone soup were optimal under the following conditions: 30% dried shrimp added, a 1:4.5 material–water ratio, and 2.7 h of stewing time. Sensory analysis showed a significant increase in the aroma, umami, kokumi, and texture intensity of the optimized sheep bone soup with dried shrimp. The untargeted approach combined with multivariate statistical analysis showed that compounds with a sweet taste (Lys and Ser), a umami taste and umami enhancement (Ala-Leu, Glu-Pro, Glu-Glu, Asp-Phe, pyroglutamic acid, and cinnamic acid), a bitter taste (Gly-Leu, Leu-Leu, Ile-Lys, and taurine), a kokumi taste (γ-Glu-Met, γ-Glu-Leu, γ-Glu-Ile, N-acetylmethionine, and N-acetylphenylalanine), a sour taste (malic acid), and a popcorn-like aroma (2-acetylthiazole) contributed significantly to the flavor enhancement of sheep bone soup. In addition, the contribution of Ac-Ser-Asp-Lys-Pro could not be ignored. These results contribute to a better understanding and improvement of the flavor qualities of sheep bone soup. Full article

Proinsulin has three distinct regions: the well-folded A- and B-chains and the dynamic disordered C-peptide. The highly conserved B-chain is a hotspot for diabetes-associated mutations, including the severe loss-of-function R(B22)Q mutation linked to childhood-onset diabetes. Here, we explore R(B22)’s role in proinsulin stability using AlphaFold-predicted structures and metadynamics simulations to achieve enhanced sampling of the free energy landscape. Our results show that R(B22) stabilizes proinsulin by interacting with N86. Substituting R(B22) with E or Q disrupts this interaction, increasing conformational flexibility. The R(B22)Q variant exhibits a flattened free energy landscape, favoring unfolded states. Additional substitutions, including Gly, Ala, Lys, Tyr, Asp, and Phe, destabilize proinsulin to varying extents by weakening hydrogen bonding. Disrupting the R(B22)–N86 interaction broadly reduces inter-chain contacts, raising the risk of aggregation-prone states. Given the link between R(B22) mutations and diabetes, our study provides crucial molecular insights into proinsulin instability. These findings highlight the role of key inter-domain (A-Chain–B-chain, B-Chain–C-peptide, and A-Chain–C-peptide) interactions in maintaining protein structures and the implications this has for understanding disease-associated proinsulin variants. Full article

Urea groups appear in many biomolecules and polymers. They have a significant impact on the properties of the materials because of their inherent strength and for their ability to participate in hydrogen bonds. Typically, in classical urea-based polymer materials, the urea groups occur in their N,N′-disubstituted state. Recently, bis-aspartates have been introduced as a novel type of hindered amine resins providing, upon crosslinking with (poly)isocyanates, the polyurea–polyaspartate thermosets (PU-ASPE) for coatings, sealants, polyelectrolytes, and other applications. These materials contain N,N′N′-trisubstituted urea linkages in their structures. However, the infrared (IR) characterization of trisubstituted urea groups has not been documented in sufficient detail. Consequently, studies on the structure of aspartate-based polyurea materials often rely on data from N,N′-disubstituted ureas, which can lead to inaccurate conclusions. This study presents a detailed evaluation of the possible urea H-bonding states, focusing on the difference between the di- and trisubstituted species. Particularly, the attributions of the IR spectra to urea-based hydrogen bonding states are presented both in neat materials and their solutions. To systematize this study, we initially focus on a simple trisubstituted urea model system, tributyl urea (3BUA), and compare its spectral response with disubstituted N-butyl-N′-cyclohexyl urea (1B1CHUA) and trisubstituted N-butyl-N′,N′-dicyclohexyl urea (1B2CHUA), to elucidate their hydrogen-bonding fingerprints. This research provides a thorough understanding of the IR response of the di- and trisubstituted urea species and their structural characteristics in urea-containing materials. Full article

There is limited information on the genetic response of tomato cultivars to pathogens. This study investigated the genetic mechanism of tomato that confers tolerance against Phytophthora infestans (Ph) and Aspergills niger (Asp) infection using different tomato accessions. Also, the study examined the effect of Ph and Asp infection on the morphology and the chromosome number of the infected tomatoes. Thirty tomato seed accessions were grown in a screen house, using a complete randomized design in triplicate, and evaluated for tolerance to both pathogens using a disease severity rating scale of 1 (highly tolerant) to 6 (highly susceptible), while chromosome assessment was performed using Carnoy’s protocol. Morphological data of 28 characteristics were collected using an IPGRI descriptor, while variance components, genetic advance (GA), and heritability were estimated for treated tomatoes and controls using the R statistical program. Accession NHT0254b, which was highly tolerant to Ph (1.00 ± 0.00) and Asp (1.33 ± 0.58), was diploid (2n = 2x = 24). The Ph-treated NHT0343a was moderately susceptible (3.67 ± 2.31), with 2n = 2x = 23, while NGB00711 was moderately susceptible (4.33 ± 1.16) with 2n = 2x = 22 after Asp treatment, indicating aneuploidy. In Ph treatment, cumulative fruit weight (CFW) had the highest environmental variance (1509.57), while number of seeds (NS) showed the highest genotypic variance (8.22). In Asp treatment, NS exhibited the highest genetic advance (2.97), while CFW had the highest phenotypic variance (754.91). Heritability estimates showed that fruit length (63.0%), the size of core (65.0%), and number of chambers (60.0%) were tolerant to Ph, while only fruit length (56.0%) was tolerant to Asp in terms of yield characteristics. Thus, tolerant accessions and traits are recommended for selection and genetic improvement. Full article

Background: Persistent splenomegaly, often an incidental finding, can originate from a number of inherited metabolic disorders (IMDs). Variants of APOE are primarily known as risk factors in terms of cardiovascular disease; however, severe dysfunction of APOE can result in a disease phenotype with considerable overlap with lysosomal storage disorders (LSDs), including splenomegaly and gross elevation of N-palmitoyl-O-phosphocholine-serine (PPCS). Methods: A case study (deep phenotyping, genetic and FACS analysis) and literature study was conducted. Results: The index patient, with a family history of early-onset cardiovascular disease, presented with splenic infarctions in a grossly enlarged spleen. The identified genetic cause was homozygosity for two APOE variants (c.604C>T, p.(Arg202Cys) and c.512G>A, p.(Gly171Asp); ε1/ε1), resulting in a macrophage storage phenotype resembling an LSD that was also present in the brother of the index patient. A FACS analysis of the circulating monocytes showed increased lipid content and the expression of activation markers (CD11b, CCR2, CD36). This activated state enhances lipoprotein intake, which eventually converts these monocytes/macrophages into foam cells, accumulating in tissues (e.g., spleen and vascular wall). A literature search identified seven individuals with splenomegaly caused by APOE variants (deletion of leucine at position 167). The combined data from all patients identified male gender, splenectomy and obesity as potential modifiers determining the severity of the phenotype (i.e., degree of triglyceride increase in plasma and/or spleen size). Symptoms are (partially) reversible by lipid-lowering medication and energy restricted diets and splenectomy is contra-indicated. Conclusions: Inherited dyslipidemic splenomegaly caused by disruptive APOE variants should be included in the differential diagnoses of unexplained splenomegaly with abnormal lipid profiles. A plasma lipid profile consistent with dysbetalipoproteinemia is a diagnostic biomarker for this IMD. Full article

Nitrogen (N) is the most important fertilizer for increasing crop production, as it is absorbed by various N transporters and metabolized by a series of enzymes. Aspartate aminotransferase (ASP) facilitates the conversion of Glu to Asp for N storage. Chinese cabbage is a typical leafy vegetable that requires a large amount of N for growth. To investigate the functions of BraASPs, 10 members of the ASP gene family in Brassica rapa (B. rapa) were identified. Phylogenetic analysis and collinearity comparisons of ASP members among B. rapa, Arabidopsis thaliana (A. thaliana), Oryza sativa (O. sativa), Brassica napus (B. napus), and Brassica oleracea (B. oleracea) were conducted to examine evolutionary associations and genome duplication events across species. Multiple cis-acting elements associated with stress responses were identified in the promoters of BraASPs, suggesting their diverse roles. Members of BraASPs were expressed in roots, stems, flowers, siliques, and leaves, with the highest expression in leaves. Their expression levels increased rapidly at 3 h under low N conditions, peaked at 6 h, and returned to low levels at 24 h. Based on transcriptomic data, BraASP1b was identified as a candidate gene in B. rapa under low N stress, localized in the nucleus and cytoplasm. Overexpression of BraASP1b in A. thaliana resulted in a higher biomass than Col-0 under low N conditions. Full article