International Journal Of Molecular Sciences[JOURNAL]
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200 papers
Skin Cells' Protection Against UVA-Induced Changes in Co-Cultured Keratinocytes-Fibroblasts' Proteome and Released Signaling Proteins by 3--Ethyl Ascorbic Acid.
Gęgotek A, Jarocka-Karpowicz I, Mucha M … +1 more , Skrzydlewska E
Int J Mol Sci · 2026 Jun · PMID 42353265 · Full text
UVA radiation affects communication between the cells that create the human skin. To prevent UVA-induced damage, there is a constant search for compounds protecting all skin cells and homeostasis in their communication....
UVA radiation affects communication between the cells that create the human skin. To prevent UVA-induced damage, there is a constant search for compounds protecting all skin cells and homeostasis in their communication. Therefore, the aim of this study was to evaluate the effect of 24 h incubation with 3--ethyl ascorbic acid (EAA; 150 µM) on the intracellular proteome of co-cultured keratinocytes and fibroblasts after UVA irradiation (total dose 15 J/cm), and on the protein profiles released into the medium by both cell types. A proteomic approach (nanoHPLC/QOrbiTrap) allowed the identification of proteins significantly modified by UVA and EAA. In keratinocytes, UVA radiation enhanced expression of pro-inflammatory and pro-proliferative/keratinizing proteins and decreased expression of antiapoptotic and antioxidant proteins, while in fibroblasts, UVA radiation induced expression mainly of pro-inflammatory proteins, simultaneously decreasing levels of proteins involved in the antioxidant response and growth factors. Increased pro-inflammatory protein and decreased growth factor levels were also observed in the medium. EAA restored the levels of these proteins compared to control cultures. The results of this study show that EAA may protect epidermal and dermal cells by reducing levels of pro-inflammatory proteins, increasing antioxidant system activity in skin keratinocytes and fibroblasts, and normalizing intercellular signaling.
Raman Spectroscopy for Probing Pathological Protein Aggregates: Potential and Perspectives for Advanced Diagnostic Applications.
Gualerzi A, Mangolini V, Forleo L … +3 more , Cabrini C, Picciolini S, Bedoni M
Int J Mol Sci · 2026 Jun · PMID 42353264 · Full text
Parkinson's disease and Alzheimer's disease are currently classified as a major global health burden, sharing a defining pathological hallmark represented by insoluble protein aggregates of α-synuclein (α-syn) and amyloi...
Parkinson's disease and Alzheimer's disease are currently classified as a major global health burden, sharing a defining pathological hallmark represented by insoluble protein aggregates of α-synuclein (α-syn) and amyloid-β (Aβ), respectively. A defining characteristic of all amyloids is a highly ordered, unbranched filamentous morphology, where individual β-strands align perpendicularly to the filament axis. Despite recent technological advances, direct observation of protein conformational changes and amyloid formation in biological samples remains a challenge as well as the quantification of pathological aggregates in liquid biopsies. This review critically recapitulates the major advances in the application of Raman spectroscopy (RS) and surface-enhanced Raman spectroscopy (SERS) in the investigation of pathological protein aggregates in neurological disorders, with a focus on α-syn and Aβ. We discuss both in vitro structural characterization and the applications to biological and clinical samples, outlining the main challenges for clinical translation, including the need for standardized protocols. Recent achievements in the use of RS and SERS on liquid biopsies and other clinical samples are paving the way for further implementation of Raman-based approaches for the diagnosis of neurodegenerative disorders.
Evaluation of CXCL12 Diagnostic Performance in Tick-Borne Encephalitis and Other Central Nervous System Infections.
Świętoń P, Słoń J, Moniuszko-Malinowska A … +2 more , Kruszewska E, Grygorczuk S
Int J Mol Sci · 2026 Jun · PMID 42353263 · Full text
CXCL12 is a chemokine acting via CXCR4 that plays a key role in inflammatory processes by regulating leukocyte migration and immune responses, making it an important focus of research in neuroinfections. In this study, t...
CXCL12 is a chemokine acting via CXCR4 that plays a key role in inflammatory processes by regulating leukocyte migration and immune responses, making it an important focus of research in neuroinfections. In this study, the CXCL12 chemokine was evaluated as a potential diagnostic marker of different neuroinfections with particular emphasis on tick-borne encephalitis (TBE). A group of 214 patients with confirmed meningitis and/or encephalitis was included and divided according to etiology into TBE, aseptic meningitis, neuroborreliosis, and purulent meningitis. CXCL12 concentration and other inflammatory parameters were measured in cerebrospinal fluid (CSF). CXCL12 levels were compared with those of controls ( = 25) and analyzed statistically. In addition, serum was used to determine albumin concentrations. CXCL12 concentrations were significantly higher in patients with neuroinfections compared to controls and showed good diagnostic performance in the overall study group (AUC = 0.791), with a more moderate diagnostic performance observed in individual etiological groups, except in the purulent meningitis group, where the effect was not statistically significant, likely due to the small sample size. CXCL12 also demonstrated some utility in differentiating between specific etiologies; however, this effect was limited. Better diagnostic performance was observed when CXCL12 was combined with pleocytosis in a composite model differentiating between the TBE group and aseptic meningitis (AUC = 0.761). The presented results indicate the role of CXCL12 in neuroinflammation while simultaneously highlighting its potential in the development of novel diagnostic approaches for viral neuroinfections. Despite higher levels in TBE, its standalone diagnostic value is limited; however, it may enhance diagnostic accuracy when combined with other markers such as pleocytosis.
A Bone-Protective Role for IFN-γ? Evidence from Genetic Association and Osteoblast Functional Assays in Postmenopausal Osteoporosis.
de Lima CAD, Souza APO, Seabra MABL … +6 more , Guaraná WL, Oliveira BMR, Crovella S, Barbosa AD, de Azevêdo Silva J, Sandrin-Garcia P
Int J Mol Sci · 2026 Jun · PMID 42353262 · Full text
Osteoporosis (OP) is a complex disease in which several immune-related genes have been identified as contributing to susceptibility and disease progression. Despite efforts to achieve functional validation, many of these...
Osteoporosis (OP) is a complex disease in which several immune-related genes have been identified as contributing to susceptibility and disease progression. Despite efforts to achieve functional validation, many of these genes, such as interferon-gamma (), remain the subject of unresolved mechanisms. The present study aimed to examine whether the -1616 (G>A, rs2069705) polymorphism was associated with postmenopausal OP. A total of 251 OP patients and 115 healthy controls were genotyped to assess the association between the -1616 (G>A, rs2069705) polymorphism and osteoporosis. To further investigate the biological role of IFN-γ in bone metabolism, human SaOs-2 osteosarcoma cells were treated with recombinant IFN-γ (2 and 100 U/mL), and calcification and cell viability were evaluated using Alizarin Red staining and the MTT assay, respectively. We found that the rs2069705 G allele was associated with an increased risk of OP (OR = 1.45, 95% CI = 1.03-2.05, = 0.03). Furthermore, serum IFN-γ levels did not differ significantly between genotype groups. In SaOs-2 cells, IFN-γ (2 U/mL) significantly increased viability ( = 0.017) and enhanced calcification in a dose-dependent manner. The rs2069705 G allele may confer susceptibility to postmenopausal OP. IFN-γ promotes osteoblast viability and mineralization at low concentrations, suggesting a potential anabolic role that warrants further investigation in human primary osteoblasts.
Complex Distribution Phenomena and Plastic Binding of Test Chemicals in Cell Culture Experiments: Exemplification by Tebufenpyrad.
Alimohammadi M, Khalidi H, Zgheib E … +8 more , Holzer AK, Bürgers N, Brochot C, Lundquist P, Magel V, Gukalova B, Liepinsh E, Leist M
Int J Mol Sci · 2026 Jun · PMID 42353261 · Full text
Biokinetic complexities (plastic sorption, protein binding, and cellular accumulation) may cause large discrepancies between nominal and biologically effective concentrations of test compounds assessed by new approach me...
Biokinetic complexities (plastic sorption, protein binding, and cellular accumulation) may cause large discrepancies between nominal and biologically effective concentrations of test compounds assessed by new approach methods (NAMs). This case study was performed to explore a generally applicable workflow that addresses biokinetic complexities in the context of NAM-based hazard testing for next-generation risk assessment (NGRA). The pesticide tebufenpyrad (TEBU) is a challenging test compound, as it (i) is hydrophobic, (ii) has an intracellular target (mitochondrial respiration), and (iii) is acting at low concentrations (susceptible to biokinetic complexities). In the newly established NeuriTox-M neurotoxicity assay, based on human dopaminergic (LUHMES) neuron cultures, TEBU showed toxic effects at 20 nM. Mass spectrometric analyses of various experimental setups showed that a large fraction (75% to >90%) of TEBU was adsorbed to plastic. This effect was strongly attenuated by albumin in the medium. Cells, cultured on plastic, were considered unsuitable to assess cellular uptake. Therefore, alternatives were explored: when cells were used as suspension cultures (3% /) in albumin-containing medium, analysis worked best. Under such conditions, the concentration ratio (cells/medium) of TEBU was around 10. Data from an in vitro distribution (VIVD) model were in good agreement with the measurements. VIVD predicted the unbound medium TEBU concentration (C) to be 2-3 orders of magnitude below the nominal concentration and the total cellular concentration to be 10-100-fold above. Standard cell culture assays showed that the medium albumin content indeed altered the TEBU toxicity threshold. More such studies are needed to embed biokinetics information into NGRA.
Barrier and Immune Modulation by ATCC PTA 6127 in Canine Epithelial and Immune Cells Under Lipopolysaccharide Challenge.
Udrea AC, Larsen KB, Bak SY … +5 more , Christensen N, Schwarzenberg A, Rekima A, Hibberd A, Shen C
Int J Mol Sci · 2026 Jun · PMID 42353260 · Full text
Coordinated responses of intestinal epithelial and immune cells are essential for maintaining barrier integrity and immune homeostasis in dogs, yet our mechanistic understanding of probiotic-derived metabolites remains l...
Coordinated responses of intestinal epithelial and immune cells are essential for maintaining barrier integrity and immune homeostasis in dogs, yet our mechanistic understanding of probiotic-derived metabolites remains limited due to reliance on non-canine experimental models, highlighting the need for studies in canine-derived systems. Here, we investigated the effects of metabolites derived from strain ATCC PTA6127 (Lr6127), delivered as a cell-free supernatant (CFS), on canine epithelial MCA-B1 cells and macrophage-like DH82 cells subjected to lipopolysaccharide (LPS)-induced inflammatory stress. Lr6127 CFS significantly reduced epithelial permeability, decreasing FITC-dextran leakage to 94.9 ± 1.9% (normalized relative to LPS-treated control, which was set as 100%) ( < 0.001), despite no detectable transcriptional changes in tight junction, adherens junction, or mucin genes. Barrier effects were instead associated with changes in markers of cellular stress responses, with heme oxygenase expression decreasing from 0.9 ± 0.1 to 0.7 ± 0.1 ( < 0.05). In DH82 immune cells, Lr6127-derived metabolites altered LPS-induced stress- and inflammation-related gene expression patterns; enhanced anti-apoptotic responses, as reflected by the increased BCL2 expression (1.4 ± 0.1 vs. 1.0 ± 0.0; < 0.01) and elevated BCL2/BAX ratios ( < 0.01); and reduced expression of pro-inflammatory mediators including IL-6 and CCL2 ( < 0.05-0.001). Proteomic analysis corroborated that Lr6127-derived metabolites reduced the abundance of inflammatory and STAT-associated signaling proteins under LPS challenge, while indicating context-dependent changes in immune-related protein profiles under resting condition. Collectively, these results suggest that Lr6127-derived metabolites improved epithelial barrier function, which was accompanied by coordinated changes in cellular stress-related and inflammatory pathways, highlighting their potential to positively influence host responses.
The Potential Role of Vitamin D in Pathogenic Variant Carriers: A Narrative Review.
Robaczyńska J, Kiljańczyk M, Maj M … +6 more , Kiljańczyk A, Byrski T, Cybulski C, Janiuk I, Gronwald J, Lubiński J
Int J Mol Sci · 2026 Jun · PMID 42353259 · Full text
Vitamin D is a fat-soluble secosteroid essential for skeletal development and calcium homeostasis, but it also exerts pleiotropic effects on numerous biological processes via its active metabolites. Vitamin D metabolites...
Vitamin D is a fat-soluble secosteroid essential for skeletal development and calcium homeostasis, but it also exerts pleiotropic effects on numerous biological processes via its active metabolites. Vitamin D metabolites act as steroid hormones that regulate cell-cycle progression, proliferation, differentiation, apoptosis, immune responses, and multiple intracellular signaling pathways. Moreover, they modulate the expression of genes involved in carcinogenesis. As circulating vitamin D levels are influenced by diet, fortified foods, and supplementation, they represent a potentially modifiable factor. Whether vitamin D status affects cancer risk or disease progression in carriers of pathogenic variants remains unclear and continues to be actively investigated. Clarifying this relationship could have significant clinical implications for risk stratification and prevention in this high-risk population. This narrative review summarizes current evidence from epidemiological, clinical, and molecular studies examining the role of vitamin D in pathogenic variant carriers. It also highlights key limitations in the existing literature and identifies critical directions for future research, emphasizing the need for well-designed prospective studies in representative cohorts.
Shared Transcriptomic Signatures and Network Interactions Between Lung Adenocarcinoma and Asthma.
Akduman S, Düz E, Gündoğdu M … +4 more , Tecimel D, Dalan AB, Bayrak ÖF, Seven D
Int J Mol Sci · 2026 Jun · PMID 42353258 · Full text
Lung adenocarcinoma (LUAD) remains the leading cause of mortality worldwide, while asthma is the most prevalent chronic disease affecting individuals of all ages. The shared airway involvement in these global health conc...
Lung adenocarcinoma (LUAD) remains the leading cause of mortality worldwide, while asthma is the most prevalent chronic disease affecting individuals of all ages. The shared airway involvement in these global health concerns results in exposure to common risk factors, suggesting a potential overlap in their genetic background. Given the roles oxidative stress and chronic inflammation play in both LUAD and asthma, we aimed to investigate similarities in their molecular mechanisms and to explore whether these shared transcriptomic signatures may prove useful for potential drug repurposing hypotheses by analyzing relevant transcriptomic datasets. This analysis identified a set of genes and co-expression interactions shared between asthma and LUAD, suggesting potential common molecular mechanisms underlying both diseases. Specifically, , , and were highlighted as potential common molecular mediators underlying both diseases, offering correlative evidence for shared pathways. These genes may represent key molecular links connecting the pathogenic processes of asthma and LUAD. The transcriptomic profiles of asthma and lung cancer datasets reveal common molecular interactions, suggesting potential shared biological mechanisms between the two diseases. These findings provide a computational framework that may guide future studies investigating therapeutic associations and possible drug-gene relationships in lung adenocarcinoma and asthma.
Dissociated Humoral and Cellular Immune Responses to Recombinant Zoster Vaccine in Myeloproliferative Neoplasms Under JAK Inhibition: A Pilot Study.
Torres-González J, O'Donnell-Cortés B, Ortiz-Flores RM … +6 more , Talarczyk DPN, Cidoncha-Morcillo B, Fariñas-Guerrero F, Rodríguez-González M, García-Delgado R, Escamilla-Sánchez A
Int J Mol Sci · 2026 Jun · PMID 42353257 · Full text
Patients with myeloproliferative neoplasms (MPN) are at increased risk of herpes zoster, particularly during Janus kinase inhibitor (JAKi) therapy, yet the immunogenicity of recombinant zoster vaccine (RZV) in this setti...
Patients with myeloproliferative neoplasms (MPN) are at increased risk of herpes zoster, particularly during Janus kinase inhibitor (JAKi) therapy, yet the immunogenicity of recombinant zoster vaccine (RZV) in this setting remains incompletely characterized. We performed a prospective pilot translational study including 18 patients with MPN and a small age-matched healthy donor group ( = 4, descriptive reference only). Samples were collected at baseline, 21 days after the first dose, and 21 days after the second dose. Humoral response was assessed by anti-varicella-zoster virus (VZV) immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA), whereas antigen-specific cellular responses were evaluated after ex vivo stimulation with recombinant VZV glycoprotein E followed by flow cytometry and cytokine quantification. IgG levels increased over time in MPN patients, while cellular responses remained limited, heterogeneous, and not consistently enhanced. Cytokine production was low and variable across time points. Overall, RZV in MPN under JAKi was associated with detectable humoral responses but limited cellular activation, supporting an apparent discordance between humoral and cellular immune readouts under the experimental conditions used.
Evaluation of the Effect of Copper Sulfate Exposure on Organs in Juvenile Rats.
Öztürk O, Yılmaz S, Okan A … +6 more , Uçar S, Kaymak E, Söylemez ESA, Ateş Ş, Bor TB, Doğanyiğit Z
Int J Mol Sci · 2026 Jun · PMID 42353256 · Full text
Copper sulphate pentahydrate is widely used in agriculture to control bacterial and fungal diseases in various crops. Despite its extensive application, limited data exist regarding its potential toxic effects on juvenil...
Copper sulphate pentahydrate is widely used in agriculture to control bacterial and fungal diseases in various crops. Despite its extensive application, limited data exist regarding its potential toxic effects on juvenile rats following early-life exposure. In addition to oxidative stress and inflammation, copper overload may also trigger cuproptosis, a recently identified copper-dependent form of regulated cell death. This study aimed to investigate the histopathological, biochemical, and molecular effects of copper sulphate exposure on major organs in juvenile rats and to elucidate the associated inflammatory and oxidative stress-related mechanisms. Male and female Sprague-Dawley rats (30-40 days old, 50-70 g) were randomly assigned to control and experimental groups. Following copper sulphate exposure, histopathological examinations were performed on major organs, including the liver, kidney, heart, lung, and reproductive tissues (testis in males and ovary in females). Immunohistochemical analyses of tumor necrosis factor-alpha (TNF-α) and nuclear factor kappa B (NF-κB) were conducted. Oxidative stress parameters, including malondialdehyde (MDA), total antioxidant status (TAS), and total oxidant status (TOS), were measured using ELISA. Gene expression levels of TNF-α and NF-κB were evaluated by quantitative real-time PCR (qRT-PCR). Copper sulphate exposure induced significant histopathological alterations in all examined tissues of both male and female juvenile rats. Biochemical findings revealed increased oxidative stress, evidenced by elevated MDA and TOS levels along with altered TAS values. Furthermore, immunohistochemical and gene expression analyses demonstrated upregulation of TNF-α and NF-κB, indicating activation of inflammatory pathways. Copper sulphate exposure leads to widespread morphological changes in juvenile rats, potentially mediated by oxidative stress and inflammation. These findings provide insight into the biological impact of early-life pesticide exposure. Further studies are warranted to clarify the underlying molecular mechanisms and to develop effective preventive or therapeutic approaches.
Extracellular Vesicles in Regenerative and Cosmetic Medicine: Safety, Clinical Effectiveness, Therapeutic Applications, and Regulatory Challenges.
Contreras C, Ariza-Donado A
Int J Mol Sci · 2026 Jun · PMID 42353255 · Full text
Extracellular vesicles (EVs), particularly small extracellular vesicles (sEVs), are lipid bilayer-delimited particles involved in intercellular communication through the transfer of proteins, lipids, and nucleic acids; m...
Extracellular vesicles (EVs), particularly small extracellular vesicles (sEVs), are lipid bilayer-delimited particles involved in intercellular communication through the transfer of proteins, lipids, and nucleic acids; many products and studies in aesthetic medicine refer to these preparations as exosomes, although endosomal origin is not always demonstrated. This review examines current evidence on the mechanisms, clinical effectiveness, safety, therapeutic applications, and regulatory challenges of EV- and sEV-based interventions, complemented by an exploratory qualitative assessment of physicians' perceptions regarding clinical implementation. A narrative review of studies indexed in Scopus and PubMed was conducted with emphasis on skin rejuvenation, hair restoration, wound healing, pigmentation disorders, and inflammatory dermatoses, and responses from 12 aesthetic physicians in Colombia were analyzed qualitatively. Available evidence suggests that EVs/sEVs may promote extracellular matrix remodeling, angiogenesis, immunomodulation, and tissue repair, with potential benefits across several aesthetic and regenerative indications. However, the literature remains heterogeneous and limited by variability in biologic sources, isolation and administration protocols, insufficient high-quality clinical trials, and unresolved regulatory issues. Reports of adverse reactions linked to unapproved products marketed as exosome-based formulations further highlight the need for stronger oversight. EVs, particularly sEVs, often referred to as exosomes in the aesthetic literature, remain a promising therapeutic platform, but safe clinical integration requires rigorous validation, technical standardization, and robust regulatory frameworks.
Ethnobotany, Phytochemistry, and Pharmacological Activities of Species in Low- and Middle-Income Countries: A Systematic Review.
Maluwa C, Zinan'dala B, Chuljerm H … +2 more , Parklak W, Kulprachakarn K
Int J Mol Sci · 2026 Jun · PMID 42353254 · Full text
species (family Lamiaceae) are among the most extensively utilized medicinal plants across low- and middle-income countries (LMICs), yet their pharmacological evidence base has not been comprehensively synthesized within...
species (family Lamiaceae) are among the most extensively utilized medicinal plants across low- and middle-income countries (LMICs), yet their pharmacological evidence base has not been comprehensively synthesized within an LMIC healthcare framework. A systematic review was conducted following PRISMA 2020 guidelines and a prospectively registered protocol (PROSPERO). Five electronic databases, PubMed, Scopus, Embase, Web of Science, and Google Scholar, were searched from January 2010 to December 2025. Studies reporting ethnobotanical, phytochemical, or pharmacological data on any species were eligible. The study selection, quality assessment and data extraction were done by two independent reviewers utilizing Rayyan software. Findings were synthesized using a narrative approach. Ninety-seven studies were included. , , and were most studied. Key bioactive constituents rosmarinic acid, eugenol, linalool, β-caryophyllene, and ursolic acid, demonstrated consistent antimicrobial [minimum inhibitory concentration (MIC): 0.31-1.25 mg/mL], antioxidant [2,2-diphenyl-1-picrylhydrazyl (DPPH) IC: 12.5-89.3 µg/mL], anti-inflammatory (35-55% edema reduction), and antidiabetic (α-glucosidase IC: 0.3-1.5 mg/mL) activities. Larvicidal efficacy exceeding 90% against spp. was demonstrated in field trials. The safety profile was broadly favorable (LD > 5000 mg/kg). species represent a pharmacologically credible and preclinically well-supported botanical resource with practical relevance for LMIC health systems, particularly in antimicrobial, antidiabetic, anti-inflammatory, and vector-control applications. To realize their therapeutic potential, future research must prioritize LMIC-contextualized randomized controlled trials, standardized phytochemical reporting, and chemotype-aware product development.
Recent Advances in TiO-Based Photocatalysis for the Treatment of Pesticide-Contaminated Wastewater: Mechanisms, Limitations, and Future Perspectives.
Tran HM, Kim T, Pham TH
Int J Mol Sci · 2026 Jun · PMID 42353253 · Full text
The discharge of pesticide residues (PRs) from agricultural activities into water bodies has raised concerns about their toxicity to humans and the ecosystem. Traditional methods such as adsorption, membrane filtration,...
The discharge of pesticide residues (PRs) from agricultural activities into water bodies has raised concerns about their toxicity to humans and the ecosystem. Traditional methods such as adsorption, membrane filtration, biological treatment, and conventional filtration usually result in incomplete removal of PRs. Currently, removal of PRs using advanced oxidation processes, particularly metal oxide-based photocatalysts, is considered a promising way. This review provides a comprehensive overview of recent advances in the photocatalytic degradation of PRs using TiO-based photocatalysts (T-BPs), the most widely investigated metal-oxide photocatalyst systems. First, we discuss the distribution, types, and negative impacts of major PRs on humans and the ecosystem. Next, we explore modification methods to enhance the properties of T-BPs, including light absorption behavior, charge separation rate, and photocatalytic degradation performance toward PRs. Afterward, this review carefully examines current challenges, such as complex water matrices, T-BP stability, energy supply for photocatalysis, and toxicity reduction. Finally, we highlight key future research directions, like the development of visible light-driven photocatalysts, enhanced mineralization efficiency, reduced secondary environmental risks, and the design of highly reliable catalyst and reactor systems for sustainable large-scale applications.
Comparative Analysis of rAAV Production from Plasmid-Encoded Versus Chromosomally Integrated rAAV Transgene in HEK293 Cells.
Toth M, Rempe A, Smesnik G … +3 more , Reithofer M, Dürauer A, Grabherr R
Int J Mol Sci · 2026 Jun · PMID 42353252 · Full text
Stable cell lines have recently achieved recombinant adeno-associated virus (rAAV) titers comparable to the standard triple transfection approach, making them a promising alternative to plasmid-based production systems....
Stable cell lines have recently achieved recombinant adeno-associated virus (rAAV) titers comparable to the standard triple transfection approach, making them a promising alternative to plasmid-based production systems. However, whether integration of the rAAV transgene into the host genome influences packaging efficiency and vector quality remains unclear. In this study, we generated stable HEK293 cell lines carrying the rAAV transgene in their genome. rAAV production was enabled by supplying the rep/cap and helper genes on two plasmids, rendering vector genome generation dependent on the chromosomally integrated transgene. Although the stable cell lines produced a 4.5-fold lower titer of viral genomes (VGs) compared to the standard triple transfection method, VG-normalized potency was four times higher. Detailed particle characterization further revealed 3-fold lower plasmid backbone DNA packaging in rAAVs produced by stable cell lines relative to triple transfection. Consistent results were obtained from mass photometry and ELISA/ddPCR analyses for the double transfection condition, while discrepancies emerged under triple transfection. These findings emphasize the importance of functional and qualitative assessments for evaluating different rAAV production approaches.
Ensemble Machine Learning- and Deep Learning-Driven Identification and Validation of Sennidin B as a Novel Dipeptidyl Peptidase-4 Inhibitor.
Ali S, Shaikh S, Lim JH … +2 more , Lee EJ, Choi I
Int J Mol Sci · 2026 Jun · PMID 42353251 · Full text
Dipeptidyl peptidase-4 (DPP-4) is a key therapeutic target for type 2 diabetes (T2D). Several synthetic anti-DPP-4 drugs are currently available for the treatment of T2D; however, the need for safe and effective therapie...
Dipeptidyl peptidase-4 (DPP-4) is a key therapeutic target for type 2 diabetes (T2D). Several synthetic anti-DPP-4 drugs are currently available for the treatment of T2D; however, the need for safe and effective therapies remains unmet due to the side effects associated with existing DPP-4 inhibitors. This study aimed to integrate structure-based and machine learning (ML)-based virtual high-throughput screening to identify natural DPP-4 inhibitors. Random forest, logistic regression, support vector machine (SVM), and multilayer perceptron (MLP) models were trained on DPP-4 IC datasets. Among these, the SVM and MLP models achieved high predictive performance, with areas under the curve of 0.928 and 0.923, respectively. Screening of a natural compound database identified 107 compounds for further analysis. Subsequent structure-based screening, using sitagliptin as a positive control, identified sennidin B and doxorubicin hydrochloride as promising candidates with strong binding affinity for DPP-4. Molecular dynamics simulations (200 ns) and MM-PBSA calculations confirmed stable interactions with DPP-4. Further, sennidin B and doxorubicin hydrochloride inhibited DPP-4 activity in a concentration-dependent manner, with estimated IC values of 39.39 and 19.78 μM, respectively. Sennidin B also reduced DPP-4 mRNA and protein expression levels in Caco-2 cells. Overall, sennidin B shows promise as a natural DPP-4 inhibitor and warrants further investigation as a potential antidiabetic agent.
Microglial Dysfunction Induced by C9ORF72 Dipeptide Repeat Proteins: Biomarker and Therapeutic Perspectives.
Sharma N, An SSA
Int J Mol Sci · 2026 Jun · PMID 42353250 · Full text
The GGGGCC hexanucleotide repeat expansion (HRE) in was recognized as the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Repeat-associated non-AUG (RAN) translation o...
The GGGGCC hexanucleotide repeat expansion (HRE) in was recognized as the most common genetic cause of amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD). Repeat-associated non-AUG (RAN) translation of the expanded repeat generated dipeptide repeat proteins (DPRs), which disrupted multiple cellular processes and contributed to neurodegeneration. Emerging evidence indicated that disease pathogenesis involved both gain-of-function (GOF) and loss-of-function (LOF) mechanisms. DPR-mediated GOF toxicity induced ribosomal dysfunction, nucleolar stress, proteostatic impairment, and neuronal injury, whereas LOF disrupted lysosomal and autophagic pathways in microglia, impairing the immune homeostasis. Neuronal injury further promoted the release of damage-associated signals that triggered secondary microglial activations and chronic neuroinflammations. This review summarized current knowledge of DPR biology, microglial dysfunction, and their contributions to disease progression in C9ORF72-associated ALS/FTD. Therapeutic strategies targeting repeated RNA, DPR productions, proteostasis, autophagy, and neuroinflammatory pathways were also discussed. In addition, the potentials of fluid biomarkers, including cerebrospinal fluid poly (GP) and blood neurofilament light chain (NfL), for diagnosis, disease monitoring, and therapeutic assessment were shown. Together, these findings provided important insights into disease mechanisms and potential avenues for improved clinical management.
Molecular Profiling and Selective Pro-Apoptotic Activity of a Pruning-Derived Leaf-Surface Extract in Colorectal Cancer Cells.
Arabi SP, Scivicco M, Parisi V … +6 more , Rosa E, De Tommasi N, Bader A, Del Vecchio V, Cacciola NA, Severino L
Int J Mol Sci · 2026 Jun · PMID 42353249 · Full text
is a medicinal and aromatic plant rich in specialised secondary metabolites, but the biomedical potential of leaf-surface metabolites recovered from pruning biomass remains poorly investigated. In this study, a pruning-d...
is a medicinal and aromatic plant rich in specialised secondary metabolites, but the biomedical potential of leaf-surface metabolites recovered from pruning biomass remains poorly investigated. In this study, a pruning-derived leaf-surface extract of was obtained by brief acetone immersion followed by -hexane partitioning. Its chemical profile was investigated by ultra-high-performance liquid chromatography coupled with high-resolution tandem mass spectrometry analysis combined with feature-based molecular networking, which revealed an enrichment in methoxylated flavonoids and pentacyclic triterpenes, including oleanane- and ursane-like derivatives. The biological activity of the extract was evaluated in HCT116 colorectal cancer cells, MDA-MB-231 triple-negative breast cancer cells, and HaCaT keratinocytes. After 24 h treatment, the extract selectively reduced HCT116 cell viability in a concentration-dependent manner, with an IC of 27.8 ± 1.049 μg/mL, whereas MDA-MB-231 and HaCaT cells were less sensitive. Mechanistic analyses in HCT116 cells showed increased early and late apoptotic populations, mitochondrial membrane depolarisation, and enhanced cleavage of caspase-9, caspase-3, and PARP. These findings indicate that a chemically profiled leaf-surface extract selectively impairs colorectal cancer cell survival by activating mitochondria-mediated apoptosis. The study also supports the valorisation of pruning-derived aromatic plant biomass as a source of bioactive natural products with potential biomedical relevance.
Engineering a Fungal Non-Reducing Polyketide Synthase with an Apparently Inactive Product-Template Domain Reveals Insights into the Catalytic Reprogramming.
Yin R, Qin Y, Liang X … +5 more , Zhai Z, Zhang M, Xu D, Zhou L, Lai D
Int J Mol Sci · 2026 Jun · PMID 42353248 · Full text
Fungal iterative non-reducing polyketide synthases (NR-PKS) contain a unique product template (PT) domain for aromatic cyclization. Among them, some NR-PKSs, such as the sorbicillin NR-PKS (SorB), have an apparently inac...
Fungal iterative non-reducing polyketide synthases (NR-PKS) contain a unique product template (PT) domain for aromatic cyclization. Among them, some NR-PKSs, such as the sorbicillin NR-PKS (SorB), have an apparently inactive PT. It is unknown what role such PT plays in NR-PKS programming. In this study, the PT domain of SorB was first dissected and engineered. Removal of the PT domain from SorB did not change the product profile, but the yield decreased. Meanwhile, a significantly lower transcriptional level of the ketoacyl synthase (KS) domain was observed in the knockout mutant (). Phylogenetic tree analysis and multiple sequence alignments revealed this PT belongs to group I (C2-C7, monocyclic ring), and mutations were found at catalytic dyad sites when compared with functional fungal PTs. However, mutating these residues back to the conserved ones did not give rise to products corresponding to a functional PT, but rendered the NR-PKS unproductive. Likewise, removal of the C-methyltransferase (CMT) domain from SorB destroyed the polyketide production. Furthermore, in an attempt to alter the methylation pattern, mutations of the key substrate-binding sites of the CMT domain were made. Site-directed mutations of the C-MT led to cessation of the polyketide production. This reveals CMT is vulnerable to engineering in a collaborating NR-PKS (SorB). These results provide additional insights for catalytic reprogramming in fungal NR-PKS.
Discovery-Driven Plasma Proteomics Identifies a Multi-Protein Signature for Amyloid PET Positivity: A Machine Learning Analysis of the Bio-Hermes Cohort.
Lamprou S, Mavromati K, Gunn-Moore FJ … +1 more , Quinn TJ
Int J Mol Sci · 2026 Jun · PMID 42353247 · Full text
Alzheimer's disease is a progressive neurodegenerative disorder in which early detection remains limited by the cost and invasiveness of positron emission tomography and cerebrospinal fluid testing. We evaluated whether...
Alzheimer's disease is a progressive neurodegenerative disorder in which early detection remains limited by the cost and invasiveness of positron emission tomography and cerebrospinal fluid testing. We evaluated whether plasma proteomic profiles could distinguish amyloid PET-positive from amyloid PET-negative individuals using the Bio-Hermes cohort. After quality control and missing-data filtering, 988 participants and 295 proteins were analysed; 31 proteins showing group differences were used for supervised classification. Random Forest, Gradient Boosting, and Neural Network models were trained across four train/test splits with repeated cross-validation and class downsampling. Amyloid-positive and amyloid-negative groups differed across a subset of proteins, with five remaining significant after false discovery rate correction. Tree-based models performed most consistently, with Random Forest and Gradient Boosting achieving AUC values of 0.79-0.81 and balanced accuracy of 0.68-0.73. Eight proteins (SERPINA1, C3, CRP, APOE4, CFH, VTN, C1QTNF5, and PON1) emerged as recurring high-importance features. These findings indicate that discovery-driven plasma proteomics can identify multi-protein signatures associated with amyloid status and can complement established single-analyte blood biomarkers by adding pathway-level information.
An Archaeal Cyclodextrin Glycosyltransferase from sp.: Characterization and Application in Starch Degradation.
Li Y, Li A, Long X … +5 more , Cao Y, Zhang A, Gu J, Ma R, Zhao G
Int J Mol Sci · 2026 Jun · PMID 42353246 · Full text
Cyclodextrin glycosyltransferase (CGTase) is a highly valuable biocatalyst in industrial starch conversion, particularly for the synthesis of cyclic oligosaccharides. In this study, a CGTase, designated CGT, was cloned f...
Cyclodextrin glycosyltransferase (CGTase) is a highly valuable biocatalyst in industrial starch conversion, particularly for the synthesis of cyclic oligosaccharides. In this study, a CGTase, designated CGT, was cloned from sp. and heterologously expressed in . The recombinant enzyme was purified and biochemically characterized. CGT exhibited maximal catalytic activity at 70 °C and pH 8.0, tolerance to metal ions and EDTA, and enhanced activity in the presence of 1 M NaCl and Ca. High-performance liquid chromatography (HPLC) and high-resolution mass spectrometry (HRMS) analyses revealed that the starch products by CGT degradation were mainly large-ring cyclodextrins (LR-CDs) with polymerization degrees of 9 to 20. Altogether, the thermostability, haloalkaliphilic, and distinctive product profile make CGT a promising biocatalyst for pharmaceutical, food, and biotechnological applications.
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