Search Results (9,752)

Background/Objectives: Melioidosis, caused by Burkholderia pseudomallei, is a severe tropical infectious disease associated with high mortality in endemic regions. Early diagnosis remains challenging because conventional diagnostic methods, including culture, serological assays, and molecular techniques, have limitations in sensitivity, specificity, processing time, and accessibility in resource-limited settings. This review evaluates current diagnostic approaches and highlights the potential of short peptide biomarkers for improving melioidosis detection. Methods: A narrative literature review was conducted using four electronic databases (PubMed, Scopus, Web of Science, and Google Scholar) covering publications from 2000 to 2024. Relevant studies were identified using predefined keywords related to melioidosis diagnostics, biomarkers, and peptide-based approaches, and were screened based on relevance to diagnostic methods and peptide biomarker development in Burkholderia pseudomallei. Results: Several biomarkers have been investigated for melioidosis diagnostics, including capsular polysaccharide (CPS), type III secretion system 1 (TTS1), and other virulence-associated proteins such as Hcp1 and BPSS1187. Among these, CPS and TTS1 are highly conserved and specific targets widely used in molecular and antigen-based detection methods. Short peptide epitopes derived from these antigens demonstrate promising advantages over whole proteins, including improved stability, high specificity, easier synthesis, and reduced production costs. Advances in epitope prediction technologies and peptide-based biosensors have further expanded the potential applications of short peptides in rapid diagnostic platforms, including ELISA, lateral flow immunoassays, and biosensor-based detection systems. Conclusions: Short peptide–based biomarkers represent a promising strategy for developing rapid, sensitive, and cost-effective diagnostic tools for melioidosis, particularly in endemic and resource-limited settings. Full article

In this study, the applicability of achiral column chromatography—including both medium-pressure liquid chromatography (MPLC) and classical gravity-driven techniques—was evaluated as a laboratory method for enantiomeric enrichment of scalemic (non-racemic) samples of axially chiral compounds. As model substrates, 3-(ortho-substituted-phenyl)quinazolin-4-one derivatives were employed. The results confirmed that self-disproportionation of enantiomers (SDE), occurring during column chromatography (SDEvCC), enabled the efficient isolation of enantiomerically pure fractions, with MPLC demonstrating particularly high effectiveness. Additionally, the parameters governing gravity-driven column chromatography were systematically optimized, with particular attention to variables such as eluent type and concentration, stationary phase composition, sample preparation protocol, and solvent purity. Furthermore, leveraging known crystallographic data and quantum chemical calculations based on Density Functional Theory (DFT), a molecular association mechanism was proposed to elucidate the physicochemical basis of the SDE phenomenon. Full article

Background/Objectives: Circulating tumour DNA (ctDNA) assays enable non-invasive assessment of tumour burden and treatment response in oncology. However, quantitative ctDNA outputs (such as variant allele frequency, tumour fraction, and aggregate burden scores) remain difficult to interpret and compare across platforms. This evidence-mapping review evaluates current quantitative reporting approaches in pancreatic ductal adenocarcinoma (PDAC) and examines the potential role of KRAS mutant ctDNA as a biologically grounded reference metric. Methods: A systematic literature search was conducted across PubMed/MEDLINE and Scopus to identify studies reporting quantitative ctDNA metrics in PDAC. Eligible studies included those measuring plasma KRAS mutations and/or reporting variant allele frequency, tumour fraction, or multi-locus aggregate metrics. Additional relevant primary studies identified through broader manual searching of PubMed were assessed against the same prespecified eligibility and classification criteria before inclusion. Data were synthesised narratively, focusing on reporting frameworks, units of measurement, assay characteristics, and the interpretability of quantitative outputs across platforms. Results: Substantial heterogeneity was observed in ctDNA quantification methods and reporting standards. Ratio-based metrics such as variant allele frequency and tumour fraction were commonly used but varied according to assay design, plasma input volume, and background cell-free DNA levels. Few studies reported absolute mutant molecule counts per unit volume. Given that approximately 90–95% of PDACs harbour truncal activating KRAS mutations, plasma KRAS was consistently represented across platforms and demonstrated potential as a shared quantitative anchor. Limited standardisation was noted in distinguishing detectability from quantifiability based on sampling depth and counting statistics. Conclusions: Current ctDNA reporting in PDAC lacks a shared quantitative reference, limiting cross-study comparability. Reporting KRAS mutant molecules per millilitre and adopting an assay-agnostic framework distinguishing detection from quantification may improve interpretability, support harmonisation across platforms, and facilitate cumulative learning in pancreatic cancer ctDNA research. Full article

Microbial contamination in aquatic environments poses severe threats to aquaculture sustainability, ecological balance and public health. Traditional culture-based detection methods, while standardized, are time-consuming and labor-intensive, often failing to meet the urgent need for rapid on-site monitoring required to prevent disease outbreaks and manage water quality effectively. By integrating latest research advances (2020–2025), this study reviews advances in rapid detection technologies for aquatic microorganisms, including the evolution of nucleic acid amplification strategies, with a focused comparison of the analytical sensitivity and field deployability of quantitative polymerase chain reaction (qPCR) and mainstream isothermal amplification techniques (loop-mediated isothermal amplification, LAMP; recombinase polymerase amplification, RPA). Furthermore, this study reports on the emergence of Clustered Regularly Interspaced Short Palindromic Repeat (CRISPR)-associated protein (Cas) systems as next-generation diagnostic tools, highlighting their integration with microfluidic Lab-on-a-Chip (LOC) platforms to achieve attomolar sensitivity. We also consider the application of portable nanopore sequencing for real-time pathogen identification and the growing role of Artificial Intelligence (AI) in analyzing complex diagnostic datasets. Advanced molecular methods have achieved significant reductions in time consumption—from days to less than one hour—while challenges regarding sample preparation and environmental matrix inhibition remain. The future of aquatic monitoring lies in integrated, automated systems that combine the specificity of CRISPR-Cas diagnostics with the connectivity of IoT-enabled biosensors. Comparative analysis indicates that isothermal amplification methods (LAMP, RPA) coupled with CRISPR-Cas systems offer the optimal balance of sensitivity, speed, and field deployability for point-of-care aquaculture diagnostics, while qPCR/dPCR remain indispensable for quantitative regulatory applications. We propose a structured technology selection framework to guide researchers and practitioners in choosing appropriate detection modalities based on specific sensitivity, cost, throughput, and deployment requirements. Full article

Background/Objectives: Sepsis is a leading cause of hospital mortality and represents a time-sensitive medical emergency. Current diagnostic strategies rely on clinical assessment, severity scores, biomarkers, and blood cultures. However, blood cultures require 24–72 h for pathogen identification and demonstrate limited sensitivity, while biomarkers such as procalcitonin and C-reactive protein lack optimal specificity. These limitations support the widespread empirical use of broad-spectrum antibiotics and highlight the need for rapid, sensitive, and culture-independent diagnostic tools. Methods: A narrative literature review was conducted using PubMed and Google Scholar, including 28 studies published over the past 10 years, encompassing observational and preclinical investigations. Current evidence on the application of Raman spectroscopy in sepsis was summarized, with a dual focus on pathogen identification and the assessment of the host response. Results: Raman spectroscopy has demonstrated the ability to detect early molecular alterations in circulating immune cells and mitochondrial redox status, potentially preceding conventional biomarkers. For pathogen identification, Raman techniques have achieved diagnostic accuracies comparable to automated systems, but with significantly shorter turnaround times. Integration with microfluidics, optical tweezers, and deep learning algorithms has further enhanced performance, although these applications remain largely experimental. Conclusions: Despite these promising results, the lack of methodological standardization, spectral overlap among phylogenetically related species, limited large-scale validation, and challenges in interpreting certain spectral signatures remain unresolved. Most available evidence originates from preclinical, single-center, and controlled studies, underscoring the need for prospective multicenter trials and harmonized protocols. Full article

Adenosine nucleotides are vital bioactive molecules with potential applications in functional foods and clinical nutrition; however, their poor membrane permeability limits their bioavailability. The utilization of plant proteins is often hindered by their poor solubility and digestibility. To address these challenges, we developed a strategy involving the formation of complexes between the walnut protein (WP) and four adenosine nucleotides. Spectroscopy, mass spectrometry, cell model, molecular docking, and other experimental techniques were conducted in this study; these methods demonstrated that such a complexation significantly enhanced the solubility of the WP to 3~4 mg/mL, while also enhancing its digestive stability in the gastrointestinal tract by 2~3-fold. Most notably, while all adenosines interacted with the protein matrix, cAMP exhibited a superior absorption efficiency, around 100-fold compared with its linear counterparts. Mass spectrometry and molecular docking were combined to reveal a new absorption mechanism for cAMP with the WP hydrolysate. These findings suggest that the complexation of WP and adenosine nucleotides offers a platform to overcome plant protein limitations and achieve efficient intracellular adenosine delivery, thereby establishing a foundation for its use in the development of functional foods. Full article

Environmental DNA (eDNA) offers a promising, non-invasive approach for monitoring infectious agents in aquaculture. While molecular techniques for detecting shrimp pathogens are well established in host tissues, there is a lack of standardized protocols for pathogen detection from environmental samples using conventional PCR. In this study, we developed and validated a universal conventional PCR protocol for monitoring DNA from major viral and bacterial shrimp pathogens within pond water and sediment samples. The method was applied to two commercial shrimp farms in Mexico, where eDNA was extracted from field-collected water and sediment. Using published primer sets, we successfully amplified DNA sequences corresponding to six key pathogens—Infectious hypodermal and hematopoietic necrosis virus (IHHNV), Baculovirus penaei (BP), Monodon baculovirus (MBV), Shrimp hemocyte iridescent virus (SHIV), Candidatus Hepatobacter penaei (NHP-B), and Acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio spp.—in environmental samples. Sequencing of PCR amplicons confirmed 93–100% identity to previously reported pathogen strains, highlighting the method’s reliability. Pathogen detection rates varied by site, sample type, and date, with the percentage of positive samples ranging from 11.1% to 77.7%. Notably, this is the first report of SHIV DNA detection from environmental samples in the Americas, highlighting its value for pathogen surveillance even in the absence of documented outbreaks. This protocol offers a cost-effective and scalable tool for pathogen surveillance in shrimp aquaculture, enhancing early disease detection and contributing to improved biosecurity and risk assessment frameworks. Full article

Hordeum brevisubulatum ‘Mengnong No. 2’ is a new forage variety developed using traditional group selection breeding techniques. It features notable advantages in plant height, tillering capacity, and overall biomass yield. However, key molecular breeding techniques such as molecular marker identification and genetic manipulation have yet to be established for this variety, limiting improvements in important traits. Consequently, we assessed the biomass of ‘Mengnong No. 2’ against ‘Mengnong No. 1’, the most widely cultivated variety in the central and western regions of Inner Mongolia, China. We report that fresh forage, dry forage, and seed yields of ‘Mengnong No. 2’ increased by 20.6%, 31.78%, and 34.35%, respectively, compared with the control variety, indicating broad prospects for its application and promotion. To enable rapid identification of ‘Mengnong No. 2’ during its promotion and to prevent production losses caused by variety admixture, we used three screened SSR primer pairs (GST25, GST37, GST127) to construct a DNA fingerprint for five H. brevisubulatum varieties, including ‘Mengnong No. 2’. With the percentage of polymorphic bands exceeding 95%, these profiles enabled precise identification of the ‘Mengnong No. 2’ variety. Furthermore, callus regeneration in H. brevisubulatum represents a bottleneck for directed molecular breeding techniques such as genetic transformation and gene editing. Accordingly, we selected the inflorescences of ‘Mengnong No. 2’ as explants and investigated the callus induction and regeneration capacity of inflorescences at different developmental stages. We found that explants at the spikelet primordia differentiation stage exhibited the highest callus induction and regeneration efficiencies, reaching 62.7% and 72.8%, respectively. The resulting embryogenic callus lines can serve as recipients for Agrobacterium-mediated transformation or gene gun bombardment, facilitating the development of subsequent high-efficiency genetic transformation and gene-editing systems. The SSR-based variety identification system and the highly efficient regeneration technology using inflorescence-derived callus established in this study lay a solid foundation for the development of a molecular breeding system for ‘Mengnong No. 2’. Full article

The species Psychotria densicostata Müll.Arg. is a shrub belonging to the Rubiaceae family, endemic to Brazil. So far, there are reports neither of phytochemical work on nor of biological evaluation of it. This study investigated its alkaloid profile and evaluated the inhibitory effects of extracts, alkaloid-enriched fractions and one of its major constituents on human neutrophil elastase (HNE). The monoterpene indole alkaloids (MIAs) strictosidine (1), (3α,5α)-5-carboxystrictosidine (2), strictosidine lactam (3), lyaloside (4), lyalosidic acid (5), 5-carboxystrictosamide (6), 3,4-dehydrostrictosidinic acid (7), and N-glucopyranosyl vincosamide (8) were characterized in mixture, in its leaves, and/or stems by using an integrated approach combining nuclear magnetic resonance (NMR) techniques, high performance liquid chromatography coupled to a tandem mass spectrometer with an electrospray ionization source (HPLC-ESI-MS/MS), and molecular networks. The crude leaf extract and an alkaloid-enriched fraction derived from it showed inhibitory activity against HNE. These results contribute to the chemical knowledge of the species and suggest its potential biological property. Full article

Siponimod, a selective sphingosine 1-phosphate (S1P) receptor modulator, represents a next-generation therapeutic drug for active secondary progressive multiple sclerosis. This study conducted in-depth forced degradation studies of siponimod in solid state subjected to acidic, alkaline, oxidative, photolytic, and thermal conditions, in compliance with ICH guidelines Q1A (R2) and Q3A (R2). An HPLC method was developed to quantify siponimod and separate its degradation products (DPs). The DPs were characterized using LC-HRMS/MS and LC-MSⁿ techniques. Moreover, the toxicological profiles of siponimod and its DPs were evaluated through the in silico tools ProTox 3.0 and ADMETlab 3.0, with molecular docking and dynamics simulations assessing their binding to the S1P1 receptor. Siponimod was stable to light but degraded under acidic, alkaline, oxidative, and thermal stress, producing five products: DP-1 (acidic), DP-2/3 (oxidative), DP-4 (hydrolytic), and DP-5 (thermal). The toxicity prediction suggested that neither siponimod nor its DPs exhibited carcinogenic or mutagenic potential, and the molecular modeling analysis revealed that DP-2 and DP-3 demonstrated favorable binding affinities, with stable dynamic profiles and thermodynamic properties that closely resembled those of siponimod. As far as we know, this is the first study on the structural elucidation of the DPs of siponimod by LC-HRMS/MS and LC-MSⁿ. Full article

Photoacoustic imaging (PAI) is an emerging hybrid biomedical imaging modality that combines the high molecular contrast of optical excitation with the deep tissue penetration of ultrasound detection. This review presents recent advances in PAI-based techniques for the detection and characterization of gynecological diseases in women, with particular focus on endometriosis and uterine-related disorders. We summarize the application of PAI across preclinical and translational studies, highlighting progress in photoacoustic microscopy, spectroscopic photoacoustic imaging, and endoscopic and probe-based implementations for non-invasive, high-resolution tissue evaluation. The role of functional and contrast-enhanced PAI approaches is discussed, emphasizing their ability to enhance diagnostic sensitivity, enable longitudinal monitoring, and provide detailed information on vascular, biochemical, and structural tissue characteristics. Furthermore, the expanding applications of PAI in assessing uterine, cervical, and ovarian pathologies, including tumor detection and tissue remodeling, are reviewed. Finally, current challenges, limitations, and future directions toward clinical translation are addressed. Collectively, this review underscores the potential of photoacoustic imaging as a powerful, non-invasive platform for early diagnosis, disease monitoring, and improved management of women’s health conditions. Full article

Accurate identification of cariogenic bacteria is crucial for understanding caries development in children. Classical culture methods often underestimate microbial diversity, while polymerase chain reaction (PCR) can detect species that are difficult to cultivate. The aim of this study was to compare culture-based and PCR-based methods for detecting key cariogenic microorganisms in the carious dentin of pediatric patients. Thirty dentin samples were collected from the permanent teeth of children aged 8–14 years. Parallel analyses were performed using standard culture techniques and PCR targeting the gtfB gene of S. mutans and the 16S rRNA gene of Lactobacillus spp. Culture results were quantified as colony-forming units, while PCR results were classified as negative, low-positive, or positive. The results show that culture-based methods identified S. mutans in 16.7% of the samples and Lactobacillus spp. in 3.3%, while PCR identified a signal for S. mutans in 43.3% and Lactobacillus spp. in 100% of the samples. PCR-based methods provide higher sensitivity for detecting key cariogenic bacteria, including S. mutans and Lactobacillus spp. However, PCR detects bacterial DNA and does not indicate bacterial viability or activity. Combining molecular and culture-based approaches allows a more comprehensive assessment of the cariogenic microbiota, supporting accurate microbiological evaluation in pediatric caries research. Full article

The evaluation of the safety of chemical substances requires the identification of a safe dose, which has no adverse effects on humans. This is obtained through animal studies, with exposure prolonged for months. Repeated-dose toxicity is a term in toxicology and pharmacology referring to the highest tested dose of a substance, so-called No Observed Adverse Effect Level (NOAEL). Experimental data on NOAEL taken from the literature and the OpenFoodTox database (total n = 848). To speed up the processing of the enormous number of substances we are exposed to, in silico models are an attractive solution. Monte Carlo technique, incorporating the Las Vegas algorithm, was applied to develop models for repeated-dose toxicity in rats. Optimal descriptors were calculated using correlation weights for attributes of the Simplified Molecular Input Line Entry System (SMILES). Computational experiments were conducted 5 times, with splits obtained using the Las Vegas algorithm. Good predictive potential was observed for these models, with an average determination coefficient on the validation set of 0.77 ± 0.04. Full article

SARS-CoV-2 infection has emerged worldwide. To reduce the number of cases and limit the transmission of the virus, health and local authorities have implemented several strategies. Mass screening is a key strategy for mitigating the damage caused by this pandemic. This strategy is based on the use of qRT-PCR and pooling to diagnose SARS-CoV-2 infection. The present work explores the performance and limitations of this strategy for the molecular diagnosis of SARS-CoV-2 infection. Three important technical aspects were retained: the comparison of two commercial extraction kits (BIGFISH and BIOER), the simulation of a non-compliant nasopharyngeal swab, and the evaluation of the pooling strategy. A total of 97 SARS-CoV-2-positive nasopharyngeal samples were used. The comparison of the two extraction kits was based on threshold cycles (Ct) values. The results showed a significant difference (IC = 95%) in the Ct of the nucleocapsid gene (N; p = 0.0000384) and RNA-dependent RNA polymerase (RdRp; p = 0.0254). However, no significant difference was observed between the Internal Control gene (IC; p = 0.0723) and Envelope gene (E; p = 0.150). The Ct values resulting from the BIGFISH extraction kit were generally lower than those obtained from BIOER. In terms of sensitivity, the RT-qPCR technique allows for the detection of viral RNA up to 10⁻³ as a dilution factor. This study demonstrated that the pooling strategy is an effective diagnostic technique. Positive samples remained detectable even in pools of 1000 or even 10,000 samples. However, the size of the pool under diagnostic conditions should not exceed a limit that must be dynamically adapted to prevalence to ensure economic and analytical viability. Full article

As essential branches of systems biology, metabolomics and lipidomics systematically reveal the composition, dynamic changes, and biological functions of small-molecule metabolites and lipids using high-throughput analytical techniques. This review examines the application of these omics technologies in evaluating livestock and poultry meat and egg quality, focusing on their roles in elucidating the molecular mechanisms behind key traits such as flavor, tenderness, and nutritional value. By identifying key metabolic markers—including glutamic acid, inosine monophosphate, and specific triglycerides—the intrinsic links between these markers and intramuscular fat deposition, flavor precursor formation, and antioxidant capacity are highlighted. Furthermore, this paper emphasizes the transformative impact of integrating multi-omics data with artificial intelligence (AI). AI-driven analytical frameworks are overcoming the limitations of traditional high-dimensional data processing, enabling robust biomarker discovery, predictive modeling for product quality, and reverse design for genetic improvement. Ultimately, the synergistic application of metabolomics, lipidomics, and AI will drive the development of modern animal husbandry toward intelligent, predictable, and precision-based production. Full article