Search Results (25,193)

We report the first synthesis of IspH-directed prodrugs targeting the terminal enzyme of the 2-C-methyl-D-erythritol 4-phosphate (MEP) pathway, (E)-4-hydroxy-3-methylbut-2-enyl diphosphate reductase (IspH or LytB). A series of alkyne and pyridine monophosphate cycloSaligenyl (cycloSal) prodrugs were prepared to enhance membrane permeability by masking the phosphate group. The effects of electron-withdrawing (Cl, CF₃) and electron-donating (OCH₃, NH₂) substituents were examined, together with amino acid-functionalized and mutual prodrug analogs. Among the synthesized compounds, chlorine-substituted derivatives 5c and 6c displayed the strongest antimycobacterial activity against Mycobacterium smegmatis, surpassing isoniazid in agar diffusion assays. These results indicate that electron-withdrawing substituents accelerate prodrug hydrolysis and facilitate intracellular release of the active inhibitor. This work provides the first experimental evidence of an IspH-targeted prodrug approach, highlighting the cycloSal strategy as a valuable tool for delivering phosphorylated inhibitors and developing novel antimycobacterial agents acting through the MEP pathway. Full article

Amino acid availability is central for the synthesis of macromolecules and numerous bioactive compounds. Amino acids are also involved in ATP production, cell signaling, and the epigenetic regulation of gene expression in human cells. From clinical and experimental studies, it appears that supplementation with specific amino acids may be relevant to correct for amino acid deficiency in the case of insufficient supply from dietary proteins with regards to the amounts needed for optimal metabolism and physiological functions. Clinical and experimental arguments suggest that amino acid supplementation may be indicated in specific situations under a specific nutritional context. However, it is essential not to overdose with excessive quantities of amino acids in supplements beyond the upper levels of safe intake (ULSI). In this narrative review, I recapitulate the protein and amino acid requirements for the general population and for subgroups of the population, and these requirements are compared to the usual consumption. Typical examples of clinical trials showing the benefits from amino acid supplementation in different physiological and pathophysiological contexts are presented together with results obtained from experimental studies. Parameters such as the no-observed-adverse-effect-level (NOAEL) values used to determine the ULSI for amino acid supplementation are defined, and values determined in clinical trials are given and discussed. Finally, prospects for future research in the field are proposed. Full article

Background: Pseudomonas aeruginosa is one of the leading agents causing healthcare-associated infections (HAIs) due to its intrinsic resistance, its capacity to acquire resistance mechanisms, and its persistence in hospital environments. In Mexico, it ranks among the most frequently reported pathogens in national surveillance systems. The aim of this study was to characterize antimicrobial resistance profiles and the genetic determinants associated with MDR/XDR phenotypes in P. aeruginosa strains from HAIs at Hospital Juárez de México (HJM). Methods: Sixty-three strains from patients with HAIs were analyzed. Identification was confirmed by 16S rRNA gene sequencing. Antimicrobial susceptibility testing followed CLSI guidelines. MDR/XDR phenotypes were classified according to the Latin American consensus for categorizing MDR, XDR, and PDR pathogens. Screening for resistance mechanisms was carried out by PCR for the main β-lactamases circulating at HJM. Finally, mutations in the oprD gene were detected in imipenem-resistant isolates through amino acid sequence alignment. Results: Resistant phenotypes allowed the identification of MDR and XDR profiles. Only the metallo-β-lactamase bla_VIM was detected. Analysis of oprD porin sequences revealed recurrent mutations (S103T, T115K, L170F, G186P, and T189V) associated with imipenem resistance. Conclusions: In P. aeruginosa, the presence of bla_VIM and structural alterations in OprD confirms the multifactorial nature of carbapenem resistance. These findings underscore the need to strengthen microbiological surveillance programs and antimicrobial stewardship strategies to mitigate the impact and spread of MDR/XDR isolates. Full article

Protoparvoviruses are highly contagious pathogens that cause severe, often fatal diseases in both domestic and wild carnivores. Golden jackal (Canis aureus) populations have experienced expansion in recent years, increasingly occupying urban and peri-urban areas. Despite this, they remain largely overlooked in scientific research. This study aimed to detect and characterise Protoparvovirus carnivoran1 circulating in a golden jackal population in Croatia and to assess their role in the epidemiology of parvovirus infections in companion animals. Small intestines from 55 jackals hunted in 2024 and 2025 were tested for Protoparvovirus carnivoran1 using real-time PCR. Positive samples were found across all sampling sites, with an overall positivity rate of 40%. Based on characteristic amino acid residues within the VP2 protein, the viruses detected in jackals were classified as feline panleukopenia virus (FPV). Phylogenetic analysis of the VP2 protein demonstrated considerable genetic diversity among strains circulating in Croatia. Additionally, a distinct group was identified, shared exclusively by Croatian domestic cats and golden jackals. Amino acid analysis revealed the novel A91T mutation, found only in jackals, and the E411Q mutation, unique to Croatian FPV strains. Structural modelling of the VP2 protein indicates that the observed mutations are located on the protein surface, within the antibody-binding site. These findings highlight the potential role of wild carnivores in parvovirus epidemiology and underscore the importance of including them in future surveillance and research efforts. Full article

Chloroplasts are key organelles in plants that carry out photosynthesis, convert light energy into chemical energy, and synthesize organic compounds. In this study, a stably heritable chlorophyll-deficient mutant was screened from the ethyl methanesulfonate-induced mutation library of Wuyunjing 21 (WYJ21). This mutant was designated as chlorophyll deficient 6 (cde6). The cde6 mutant exhibits a low chlorophyll content, photosynthetic defects, an impaired chloroplast structure, a significant reduction in the number of stacked thylakoid layers, and a yellow-green leaf phenotype in the early tillering stage. Through MutMap analysis, it was found that the cde6 mutant harbors a single-base mutation (T→A) in the LOC_Os07g38300 gene. This mutation results in an amino acid substitution from valine (Val) to aspartic acid (Asp) in the encoded protein, thereby affecting the protein’s structure and function. The mutation of CDE6 leads to decreased expression of genes related to chloroplast development and chlorophyll biosynthesis. Further studies revealed that the CDE6, a potential chloroplast ribosome recycle factor, leads to high temperature sensitivity in rice when mutated. As high-temperature stress is a primary constraint to global rice productivity, the identification of CDE6 provides a genetic target for improving thermotolerance. In conclusion, these findings demonstrate that CDE6 plays a crucial role in chloroplast biogenesis and provide new insights into its regulatory function in high-temperature tolerance. Full article

Background: Solenopsis invicta, commonly known as the red imported fire ant (RIFA), is an important global invasive pest, and its management is challenging because of insecticide resistance and environmental problems. Methods: In this research, we applied transcriptomics to analyze the molecular responses of S. invicta worker ants exposed to different types of pesticides, beta-cypermethrin (BC) and the entomopathogenic fungus Cordyceps cicadae (CC), as well as to different concentrations of these pesticides. Results: A total of 2727 differentially expressed genes (DEGs) were identified across all samples. The number of DEGs in the BC treatment group was significantly higher than that in the CC treatment group (2520 vs. 433), and higher concentrations resulted in more DEGs (an increase of 47 in the BC group and 229 in the CC group). KEGG pathway analysis revealed that the DEGs were significantly enriched in lipid metabolism, carbohydrate metabolism, amino acid metabolism, signal transduction, and membrane transport. Immune-related gene analysis showed more general down-regulation (average FPKM value in BC 741.37 to 756.06 vs. CK 1914.42) of pathogen recognition genes (PGRP-SC2) under BC stress conditions, while CC treatment resulted in increases in expression of important immune effectors such as various serine proteases. Conclusions: Overall, this study provides useful insights into the molecular basis of responses to different pesticides in S. invicta and offers a basis to develop new approaches to control this pest. Full article

The misuse of antibacterial agents has contributed to the growing prevalence of antibiotic resistance, highlighting an urgent need to explore alternative anti-infection therapeutic strategies. Antimicrobial peptides (AMPs) are naturally occurring molecules. They exhibit broad-spectrum antimicrobial activity and represent promising candidates for the development of novel therapeutics. A cysteine-rich antimicrobial peptide was identified and characterized from the genome of Tigriopus japonicus and designated “TjRcys1”. The precursor form of TjRcys1 comprises 96 amino acids. Structural analyses of TjRcys1 revealed random coils, two α-helices, and two β-strands. Recombinant TjRcys1 had inhibitory effects upon Staphylococcus aureus and Bacillus sp. T2, with a minimum inhibitory concentration of 64 μM for both. TjRcys1 did not show complete inhibition against Vibrio alginolyticus, Klebsiella pneumoniae, or Aeromonas hydrophila at 64 μM, but it did slow their growth rate. TjRcys1 could disrupt the permeability of the cell membrane of S. aureus. Transcriptomic analyses indicated that TjRcys1 could interfere with the ribosome biosynthesis and nucleotide metabolism of K. pneumoniae. Our results provide a valuable reference for the development of new AMPs and optimization of their design. Full article

Several studies have demonstrated that methionine supplementation in fish diets enhances immune status, inflammatory response, and resistance to bacterial infections by modulating for DNA methylation, aminopropylation, and transsulfuration pathways. However, the immunomodulatory effects of methionine in viral infections remain unexplored. This study aimed to evaluate the effect of methionine supplementation on immune modulation and resistance to the viral haemorrhagic septicaemia virus (VHSV) in rainbow trout (Oncorhynchus mykiss). Two diets were formulated and fed to juvenile rainbow trout for four weeks: a control diet (CTRL) with all nutritional requirements, including the amino acid profile required for the species, and a methionine-supplemented diet (MET), containing twice the normal requirement of DL-methionine. After feeding, fish were bath-infected with VHSV, while control fish were exposed to a virus-free bath. Samples were collected at 0 (after feeding trial), 24, 72, and 120 h post-infection for the haematological profile, humoral immune response, oxidative stress, viral load, RNAseq, and gene expression analysis. In both diets, results showed a peak in viral activity at 72 h, followed by a reduction in viral load at 120 h, indicating immune recovery. During the peak of infection, leukocytes, thrombocytes, and monocytes migrated to the infection site, while oxidative stress biomarkers (superoxide dismutase glutathione S-transferase, and glutathione redox ratio) suggested a compromised ability to manage cellular imbalance due to intense viral activity. At 120 h, immune recovery and homeostasis were observed due to an increase in the amount of nitric oxide, GSH/GSSG levels, leukocyte replacement, monocyte influx, and a reduction in the viral load. When focusing on the infection peak, gene ontology (GO) analysis showed several exclusively enriched pathways in the skin and gills of MET-fed fish, driven by the upregulation of several key genes. Genes involved in recognition/signalling, inflammatory response, and other genes with direct antiviral activity, such as TLR3, MYD88, TRAF2, NF-κB, STING, IRF3, -7, VIG1, caspases, cathepsins, and TNF, were observed. Notably, VIG1 (viperin), a key antiviral protein, was significantly upregulated in gills, confirming the modulatory role of methionine in inducing its transcription. Viperin, which harbours an S-adenosyl-L-methionine (SAM) radical domain, is directly related to methionine biosynthesis and plays a critical role in the innate immune response to VHSV infection in rainbow trout. In summary, this study suggests that dietary methionine supplementation can enhance a more robust fish immune response to viral infections, with viperin as a crucial mediator. The improved antiviral readiness observed in MET-fed fish underscores the potential of targeted nutritional adjustments to sustain fish health and welfare in aquaculture. Full article

This review examines the feasibility of electrochemical synthesis of poly-L-arginine (PArg) using repetitive cyclic voltammetry in neutral aqueous phosphate-buffered saline. Previous studies on electrochemical deposition of PArg onto different carbonaceous electrode materials are discussed with respect to the already reported mechanistic models. Some controversial interpretations are of interest, predominantly the formation of peptide bonds during the electropolymerisation of L-arginine. Several alternative anodic pathways are considered via the possibilities and limitations of ways of attaching L-arginine molecules to the electrode surface. Furthermore, the role of oxygen-containing surface groups is discussed, as this aspect has been largely overlooked in the context of L-arginine deposition, despite the O-terminating character of the electrode surface and its effect on the reactivity of the nucleophilic guanidine group in L-arginine. Also, the application of extremely high potentials around +2 V vs. Ag/AgCl/3 mol L⁻¹ KCl is considered, as it can lead to the generation of reactive oxygen species that may interfere with or even govern the entire deposition process. Thus, the absence of such considerations may raise doubts about the peptide nature of the electrochemically assisted polymerisation of this basic amino acid. Finally, it seems that the identity of the electrochemically synthesised PArg does not correspond to that of this polymer prepared by conventional methods, such as solid-phase peptide synthesis, solution-phase synthesis, or N-carboxy-anhydride polymerisation, and therefore the whole process remains unproved. Full article

The Sanjiang Plain hosts the largest freshwater wetland in Northeastern China and plays a critical role in regional climate stability. However, climate change and human activities have degraded the wetland, forming a successional gradient from the original flooded wetland to dry shrub and forest vegetation with a lower ground water level. This degradation has altered soil microbial structure and functions, reducing ecological and socio-economic benefits. Along this successional gradient, we used Biolog-ECO plates combined with soil enzyme assays (catalase, urease, sucrase, and acid phosphatase) to assess the dynamics of microbial carbon metabolic activity, measured by average well color development (AWCD). The results showed a systematic decline in AWCD values with advancing succession, revealing a pronounced reduction in overall microbial metabolic activity during wetland degradation. This trend correlated with loss of soil moisture, organic carbon, and nitrogen nutrients. Microbial communities in early successional wetland stages (i.e., original natural wetland and wetland edge) preferred labile carbon sources (e.g., carbohydrates, amino acids), while forested stages favored relatively more structural (e.g., polymers, phenolic compounds). These findings indicate that vegetation succession regulates microbial carbon metabolism by modifying soil physicochemical properties, providing insights for wetland restoration. Full article

Edible flowers have garnered increasing attention due to their high content of bioactive compounds, making them promising candidates for biomedical and functional food applications. This work evaluated the metabolomic data of fresh Rosa x hybrida petals, revealing seven types of metabolites, including amino acids, organic acids, vitamins, sugars, phenolic acids, and flavonoids. Notably, quercetin, kaempferol and their derivatives were the main flavonoids determined. Furthermore, in vitro studies were conducted to evaluate the potential antiproliferative effects against triple-negative breast cancer (TNBC). Thus, the methanolic extract derived from Rosa x hybrida petals demonstrated significant antitumoral activity against both sensitive and resistant TNBC cells, as evidenced by reduced MTT metabolization, colony formation, and wound healing activity. Furthermore, the cell death mechanism associated with the petal extract was studied. The antiproliferative activity was mediated by reactive oxygen species generation, triggering cell death mechanisms such as apoptosis and autophagy. In conclusion, these results propose Rosa x hybrida could be a new tool for nutraceuticals and functional food production. Full article

Background: Recombinant human arginase (rhArg) has been proven to exhibit an anticancer effect via arginine starvation. To further improve the efficacy of rhArg, we examined the feasibility of a combination strategy with Bcl-2 inhibitors (ABT263 and ABT199) or an antidiabetic drug (metformin) and investigated the mechanistic basis for these strategies. Methods: The combination effects were evaluated in a panel of human cancer cell lines modeling pancreatic ductal carcinoma (PDAC), triple-negative breast cancer (TNBC), colorectal cancer (CRC) and glioblastoma (GBM). Western blot analysis was used to evaluate the expression of apoptotic and cell cycle markers. MTT assay was used to evaluate the combination efficacy. Flow cytometric assays were used to investigate the apoptotic and cell cycle effects. Results: The combination of rhArg with sublethal doses of ABT263 significantly induced dose-dependent apoptosis, with elevated expression of apoptotic markers and a CI of 0.47 in U251. The combination inhibited CDK2 and cyclin A expression, indicating that the observed synergy also resulted from cell cycle arrest. We also found that rhArg + metformin was synergistic in a time-dependent manner. Compared to other amino acid depletion agents, rhArg + ABT263 was the most favorable combination pair. Conclusions: The combination of rhArg and ABT263 enhanced apoptosis and cell cycle arrest, demonstrating a potential broad-spectrum antitumor strategy. Full article

Macrobrachium nipponense is an important commercial freshwater prawn species in China. Since larger individuals command higher market value, there is a pressing need to identify growth-related genes and single-nucleotide polymorphisms (SNPs) to facilitate genetic improvement in this species. Previous studies have suggested a potentially regulatory role of an actin-like protein (ACTL) in the growth of M. nipponense. Therefore, the present study aimed to functionally characterize the role of ACTL in growth and identify growth-associated SNPs within this gene. The open reading frame of Mn-ACTL is 1131 bp, encoding a protein with 377 amino acids. Blastx and phylogenetic analyses indicated that Mn-ACTL shares a close evolutionary relationship with orthologs from Macrobrachium rosenbergii and Palaemon carinicauda. The highest expression level of Mn-ACTL in muscle tissue detected by qPCR suggested its potential involvement in growth regulation. RNA interference experiments showed that prawns injected with dsGFP exhibited larger body sizes than those injected with dsACTL, indicating that knockdown of Mn-ACTL expression inhibits growth performance in M. nipponense. Furthermore, muscle tissue from the dsACTL-injected group displayed looser myofibril packing, visibly eroded areas, and increased sarcomere spacing. Collectively, these results demonstrated that ACTL positively regulates growth in M. nipponense. Additionally, the T allele at locus S28_17149891 and the G allele at locus S28_17145758 were significantly associated with growth traits (p < 0.05). In conclusion, this study confirmed the positive regulatory role of ACTL in growth and identified growth-associated SNPs in M. nipponense, providing valuable insights for breeding new varieties with enhanced growth performance in this species. Full article

The family Capitellidae performs critical roles in bioturbation and sediment remediation within global marine benthic ecosystems. However, they are a taxonomically challenging group due to their simple morphology and a ‘morphological mosaic’, where traditional classificatory traits, such as thoracic chaetiger counts, appear convergently across genera. Previous multi-locus studies (using 18S, 28S, H3, and COI) first highlighted this conflict, revealing the polyphyly of major genera like Notomastus and even Leiochrides itself (based on unidentified specimens). More recently, mitogenomic studies uncovered massive gene order rearrangements and a conflicting topology but did not include Leiochrides. Critically, with no complete mitogenome reported for a formally identified Leiochrides species, its true phylogenetic position and the validity of its polyphyly remain unresolved. To address this critical gap, we sequenced and characterized the first complete mitochondrial genome from a formally identified species, Leiochrides yokjidoensis, recently described from Korean waters. The complete mitogenome was 17,933 bp in length and included the typical 13 protein-coding genes (PCGs), 2 ribosomal RNAs (rRNAs), and 22 transfer RNAs (tRNAs). Gene order (GO) analysis revealed the occurrence of gene rearrangements in Capitellidae and in its sister clade, Opheliidae. A phylogenomic analysis using the amino acid sequences of 13 PCGs from 30 species established the first robust systematic position for the genus Leiochrides (based on this formally identified species). Phylogenetic results recovered Leiochrides as a sister group to the clade comprising Mediomastus, Barantolla, Heteromastus, and Notomastus hemipodus (BS 99%). This distinct placement confirms that Leiochrides represents an independent evolutionary lineage, phylogenetically separate from the polyphyletic Notomastus complex, despite their morphological similarities. Furthermore, our analysis confirmed the polyphyly of Notomastus, with N. hemipodus clustering distinctly from other Notomastus species. Additionally, signatures of positive selection were detected in ND4, and ND5 genes, suggesting potential adaptive evolution to the subtidal environment. This placement provides a critical, high-confidence anchor point for the genus Leiochrides. It provides a reliable reference to investigate the unresolved polyphyly suggested by previous multi-locus studies and provides compelling evidence for the hypothesis that thoracic chaetiger counts are of limited value for inferring phylogenetic relationships. This study provides the foundational genomic cornerstone for Leiochrides, representing an essential first step toward resolving the systematics of this taxonomically challenging family. Full article

Cardiac metabolic flexibility is a key determinant of myocardial energetic resilience. In heart failure with reduced ejection fraction (HFrEF), intrinsic mitochondrial dysfunction and lipotoxicity compromise oxidative capacity. In contrast, heart failure with preserved ejection fraction (HFpEF) is orchestrated primarily by systemic comorbidities and coronary microvascular dysfunction, which decouple glycolysis from glucose oxidation. This review integrates these distinct pathophysiologies into a comprehensive biomarker framework. Beyond core hemodynamic markers, we detail indices of metabolic flux (ketones, acylcarnitines, branched-chain amino acids), endothelial injury, and fibrosis. We further prose a shift from static, isolated measurements to dynamic functional profiling using standardized challenges (e.g., mixed-meal or exercise tests) to quantify metabolic suppression and recovery kinetics. This structured hierarchy enables phenotype-tailored risk stratification and guides mechanism-based precision therapies in the era of personalized medicine. Full article