Current Issues in Molecular Biology

15 pages, 1557 KB

Open AccessArticle

Tea Polysaccharides Ameliorates Non-Alcoholic Fatty Liver Disease in Mice via Regulating Macrophages Polarization by Gut Microbial Metabolites

by Daixin Liu, Ang Li and Ping Li

Curr. Issues Mol. Biol. 2026, 48(3), 338; https://doi.org/10.3390/cimb48030338 - 23 Mar 2026

Viewed by 328

Abstract

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and a global public health concern, for which there is currently no effective method to inhibit its progression. The pathogenesis of NAFLD is related to hepatic lipid metabolism disorders and liver [...] Read more.

Non-alcoholic fatty liver disease (NAFLD) is the most common chronic liver disease and a global public health concern, for which there is currently no effective method to inhibit its progression. The pathogenesis of NAFLD is related to hepatic lipid metabolism disorders and liver inflammation. Previous studies have shown that tea polysaccharides (TPS) have the ability to regulate lipid metabolism and control inflammation. This study aimed to observe the effect of TPS on ameliorating NAFLD in a mouse model and to reveal its underlying mechanisms. In the current study, male C57BL/6J mice were fed a high-fat diet and administered 100 mg/kg TPS daily by gavage for 14 weeks. Then, liver injury indicators and macrophage polarization markers were detected. The results revealed that TPS could significantly ameliorate the progression of NAFLD and decrease liver injury indicators. Moreover, we found that treatment of NAFLD model mice with TPS could skew liver macrophages polarization from M1 to M2 type, which inhibited pro-inflammatory cytokines production and liver inflammation. Mechanistically, TPS cannot directly regulate the polarization of liver macrophages, but instead promotes the production of butyric acid by gut microbiota, which in turn regulates macrophage polarization. These findings suggest that TPS ameliorates NAFLD-associated inflammation by modulating the gut–liver axis and promoting M2 macrophage polarization, laying the foundation for the potential of TPS in the development of health foods for NAFLD. Full article

(This article belongs to the Section Bioorganic Chemistry and Medicinal Chemistry)

► Show Figures

Figure 1

20 pages, 5800 KB

Open AccessArticle

The Mechanism of Action of Stigmasterol in Bone Formation in Osteoporosis

by Cailian Lu, Hong Li, Zhengbo Liu, Sirui Lü and Junxing Liu

Curr. Issues Mol. Biol. 2026, 48(3), 337; https://doi.org/10.3390/cimb48030337 - 23 Mar 2026

Viewed by 466

Abstract

Osteoporosis (OP) is a metabolic bone disease characterized by reduced bone mass and impaired bone microarchitecture, significantly impacting patients’ quality of life. Stigmasterol (STG), a natural plant sterol, has been reported to possess multiple biological activities. However, its effects on OP bone formation [...] Read more.

Osteoporosis (OP) is a metabolic bone disease characterized by reduced bone mass and impaired bone microarchitecture, significantly impacting patients’ quality of life. Stigmasterol (STG), a natural plant sterol, has been reported to possess multiple biological activities. However, its effects on OP bone formation and underlying molecular mechanisms remain unclear. The effects of STG on OP bone formation and potential mechanisms were investigated through in vivo and in vitro experiments combined with network pharmacology analysis. An OP model was established in ovariectomized (OVX) rats, and the bone-protective effects of STG were evaluated via micro-CT analysis and histological staining. In vitro experiments, MC3T3-E1 pre-osteoblasts were used to assess STG’s influence on osteogenic differentiation through Western blot analysis and ALP/ARS staining. Network pharmacology methods were used to predict potential targets and signaling pathways for STG in OP treatment, followed by mechanism validation. STG significantly improved bone microarchitecture in OVX rats, increased key osteogenic marker expression, and promoted MC3T3-E1 osteogenic differentiation in a dose-dependent manner. Network pharmacology analysis predicted 278 potential targets for STG in treating OP, with pathway enrichment analysis indicating significant involvement of the JAK/STAT pathway. Mechanistic studies revealed that STG promotes osteogenic differentiation by activating the JAK2/STAT3 signaling cascade. As an osteogenic promoter, STG effectively alleviates bone loss and enhances osteoblast differentiation by activating the JAK2/STAT3 signaling pathway. Full article

(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)

► Show Figures

Figure 1

20 pages, 3760 KB

Open AccessArticle

Anti-Biofilm Activity of Combinations of Cinnamic Acid and Its Derivatives with Cloxacillin Against Methicillin-Resistant Staphylococcus epidermidis

by Tomasz Zawiła, Denis Swolana, Marta Zawiła, Zuzanna Rzepka and Robert D. Wojtyczka

Curr. Issues Mol. Biol. 2026, 48(3), 336; https://doi.org/10.3390/cimb48030336 - 23 Mar 2026

Viewed by 501

Abstract

Staphylococcus epidermidis (S. epidermidis) poses a significant clinical challenge, particularly in the context of biofilm-associated infections, with increasing antibiotic resistance further complicating infection eradication. In the present study, the effects of cinnamic acid and its derivatives (ferulic acid, p-coumaric acid, [...] Read more.

Staphylococcus epidermidis (S. epidermidis) poses a significant clinical challenge, particularly in the context of biofilm-associated infections, with increasing antibiotic resistance further complicating infection eradication. In the present study, the effects of cinnamic acid and its derivatives (ferulic acid, p-coumaric acid, and sinapic acid), alone and in combination with the β-lactam antibiotic cloxacillin, on biofilm formation by a single methicillin-resistant S. epidermidis (MRSE) clinical strain were explored. The expression of the biofilm-associated icaADBC operon genes and the icaR repressor gene was assessed using Real-Time PCR as an exploratory analysis under sub-minimal inhibitory concentrations (sub-MICs) of the tested compounds. Furthermore, confocal microscopy was used to qualitatively assess selected structural changes in the biofilm. Their occurrence was demonstrated depending on the fractional inhibitory concentration (FIC) levels used. The results revealed variable and nonlinear patterns of gene expression in response to the tested concentrations. Additionally, compound-dependent differences in anti-biofilm-related responses were observed. Overall, the findings provide insight into the potential influence of cinnamic acid derivatives combined with cloxacillin on biofilm-associated processes in S. epidermidis. Full article

(This article belongs to the Special Issue Antimicrobial or Antibiofilm Activity of Biodegradable Substances)

► Show Figures

Figure 1

23 pages, 3418 KB

Open AccessArticle

Biotransformation of Maclekarpine E in Rats: CYP2C19-Mediated Metabolism, Fecal Enrichment, and Network Pharmacology-Based Anti-Ulcerative Colitis Prediction

by Yingxue Yang, Lin Wang, Jiaojiao Xue, Zhen Dong and Pi Cheng

Curr. Issues Mol. Biol. 2026, 48(3), 335; https://doi.org/10.3390/cimb48030335 - 23 Mar 2026

Viewed by 284

Abstract

Maclekarpine E is a minor alkaloid from Macleaya species with reported in vitro anti-inflammatory activity, but its in vivo metabolism remains unexplored. This study investigated the metabolic fate of maclekarpine E in rats and evaluated the potential pharmacological relevance of its metabolites. Maclekarpine [...] Read more.

Maclekarpine E is a minor alkaloid from Macleaya species with reported in vitro anti-inflammatory activity, but its in vivo metabolism remains unexplored. This study investigated the metabolic fate of maclekarpine E in rats and evaluated the potential pharmacological relevance of its metabolites. Maclekarpine E was orally administered to male Sprague-Dawley rats (250 mg/kg). Plasma, urine and feces were collected and analyzed by UPLC-Q-TOF-MS/MS. CYP phenotyping was performed using recombinant human enzymes. Molecular docking against ABCG2 and ABCC2 was conducted to assess potential interactions of all fecal compounds with these efflux transporters. Network pharmacology was employed to predict potential anti-ulcerative colitis-related targets of the metabolites, generating hypotheses for future experimental validation. Nineteen phase I metabolites were identified. Biotransformations included ring-opening, demethylation and oxidation. All 19 metabolites were detected in feces, nine in plasma and two in urine. No phase II conjugates were observed. CYP2C19 was the only significantly active isoform under the tested conditions, mediating approximately 16.5% substrate depletion (p < 0.05). All 20 fecal compounds bound ABCG2 (ΔG < −5.0 kcal/mol); 19 bound ABCC2. Network pharmacology yielded 57 overlapping targets with ulcerative colitis, enriched in PI3K-Akt and MAPK pathways. This study provides the first comprehensive metabolic profile of maclekarpine E in rats. The compound undergoes CYP2C19-mediated oxidation and is predominantly excreted into feces. Its fecal metabolites are potential ABCG2/ABCC2 substrates and may target UC-associated pathways based on network pharmacology predictions, warranting further experimental validation. Full article

(This article belongs to the Special Issue Natural Products in Biomedicine and Pharmacotherapy, 2nd Edition)

► Show Figures

Figure 1

48 pages, 8443 KB

Open AccessReview

Neurotransmitter Systems in Alzheimer’s Disease

by María Jesús Ramírez-Expósito, Cristina Cueto-Ureña and José Manuel Martínez-Martos

Curr. Issues Mol. Biol. 2026, 48(3), 334; https://doi.org/10.3390/cimb48030334 - 22 Mar 2026

Viewed by 815

Abstract

Alzheimer’s disease (AD), the leading cause of global dementia, is a multifactorial process that goes beyond the accumulation of β-amyloid (Aβ) plaques and tau protein tangles, including glia cell-mediated neuroinflammation, vascular dysfunction, metabolic alterations, and synaptic loss. Its complex etiology also involves oxidative [...] Read more.

Alzheimer’s disease (AD), the leading cause of global dementia, is a multifactorial process that goes beyond the accumulation of β-amyloid (Aβ) plaques and tau protein tangles, including glia cell-mediated neuroinflammation, vascular dysfunction, metabolic alterations, and synaptic loss. Its complex etiology also involves oxidative stress and mitochondrial dysfunction. Multiple neurotransmitter systems involved in the pathogenesis and the various cognitive and non-cognitive symptoms of AD are thus altered. The cholinergic system, historically the first to be associated with AD, suffers early degeneration and loss of neurons/receptors, correlating with cognitive impairment. The glutamatergic system, the main excitatory system, exhibits excitotoxicity due to increased extracellular glutamate and alterations in NMDA/AMPA receptor distribution, exacerbating neuronal damage. The GABAergic system, the main inhibitor, shows alterations in parvalbumin-positive interneurons, leading to hyperexcitability and dysfunction of neuronal networks. Monoaminergic systems (serotonergic, dopaminergic and noradrenergic) undergo early degeneration in key nuclei such as the raphe and locus coeruleus, contributing to the apathy, depression and sleep disturbances characteristic of AD. Other less explored systems, such as histaminergic and purinergic, are also crucial in cognitive modulation and neuroinflammation. The endocannabinoid system acts as a master modulator with neuroprotective and anti-inflammatory effects. These systems do not operate in isolation; their complex interactions generate pathological circuits that amplify neuronal dysfunction. The limited efficacy of current therapies, which are primarily symptomatic, highlights the need for multimodal approaches that may transform AD treatment toward personalized and more effective interventions. Full article

(This article belongs to the Special Issue Advances in Bioinformatics and Molecular Research on Alzheimer’s Disease)

► Show Figures

Figure 1

21 pages, 8614 KB

Open AccessArticle

Eupatorium lindleyanum DC. Suppresses Cytokine Storm by Inhibiting NF-κB and PI3K–Akt Signaling in Sepsis-Associated and Virus-Related Acute Lung Injury

by Chen Luo, Peilin He, Yan Yang, Lian Xia, Wenjie Xu, Daike Zou, Yiduo Feng, Lian Duan, Junjie Deng, Yong Jing and Xianqin Luo

Curr. Issues Mol. Biol. 2026, 48(3), 333; https://doi.org/10.3390/cimb48030333 - 21 Mar 2026

Viewed by 373

Abstract

Cytokine storm is a central pathogenic mechanism underlying sepsis-induced acute lung injury (SALI) and severe coronavirus disease 2019 (COVID-19), yet effective therapeutic strategies remain limited. Eupatorium lindleyanum DC. (EL), a traditional Chinese medicinal herb, has been reported to possess anti-inflammatory, antioxidant, and antiviral-related [...] Read more.

Cytokine storm is a central pathogenic mechanism underlying sepsis-induced acute lung injury (SALI) and severe coronavirus disease 2019 (COVID-19), yet effective therapeutic strategies remain limited. Eupatorium lindleyanum DC. (EL), a traditional Chinese medicinal herb, has been reported to possess anti-inflammatory, antioxidant, and antiviral-related activities; however, its protective mechanisms in SALI and virus-associated inflammatory lung injury remain incompletely understood. In this study, an integrated strategy combining computational prediction and experimental validation was employed to investigate the therapeutic potential and underlying mechanisms of EL. The chemical constituents of EL were characterized by UPLC–Q–TOF/MS, followed by network pharmacology, molecular docking, and molecular dynamics analyses to predict key targets and signaling pathways. A cecal ligation and puncture (CLP)-induced SALI rat model was used to evaluate lung histopathology, pulmonary edema, cytokine production, and inflammatory signaling activation. In parallel, LPS-stimulated RAW264.7 macrophages were used to assess cytokine secretion and pathway regulation in vitro. In addition, a SARS-CoV-2 pseudovirus-induced mouse model was employed to further evaluate the in vivo relevance of the representative bioactive compound hyperoside in pseudovirus-associated lung injury. A total of 32 active compounds and 697 putative targets were identified, among which 116 were associated with sepsis and COVID-19. In vivo, EL markedly alleviated lung injury, reduced the lung coefficient and wet/dry ratio, and suppressed excessive production of proinflammatory cytokines and activation of key signaling proteins. In vitro, EL dose-dependently inhibited TNF-α and IL-6 secretion and regulated the PI3K–Akt and NF-κB signaling pathways. Notably, hyperoside showed favorable predicted interactions with PI3K–Akt pathway-related targets (EGFR, PI3K, and Akt), while molecular dynamics simulations supported stable interactions with several COVID-19-related targets, including ACE2, Mpro, and RdRp. Furthermore, hyperoside significantly alleviated SARS-CoV-2 pseudovirus-associated lung injury, reduced ACE2 protein expression, and downregulated EGFR, PI3K, and Akt mRNA levels in vivo. Collectively, these findings indicate that EL exerts protective effects through multi-component, multi-target, and multi-pathway mechanisms, and support its potential value for further investigation in SALI and virus-associated inflammatory lung injury. Full article

(This article belongs to the Special Issue Advances in Phytochemical Research: Molecular Pathways in Health and Disease)

► Show Figures

Figure 1

26 pages, 1342 KB

Open AccessEditor’s ChoiceReview

Current and Developing Therapeutics for Dry Eye Disease: Targeting Ion Channels

by Rebecca Jung, Emily Kao, Victor H. Guaiquil, Ali R. Djalilian and Mark I. Rosenblatt

Curr. Issues Mol. Biol. 2026, 48(3), 332; https://doi.org/10.3390/cimb48030332 - 21 Mar 2026

Viewed by 613

Abstract

Dry eye disease (DED) is an ocular surface disorder characterized by tear film instability, inflammation, epithelial damage, and neurosensory abnormalities. Due to its multifactorial etiology and pathophysiology, conventional therapies that focus on lubrication and immunosuppression often fall short in addressing the neuropathic component [...] Read more.

Dry eye disease (DED) is an ocular surface disorder characterized by tear film instability, inflammation, epithelial damage, and neurosensory abnormalities. Due to its multifactorial etiology and pathophysiology, conventional therapies that focus on lubrication and immunosuppression often fall short in addressing the neuropathic component of ocular pain experienced by a growing subset of patients. Recent developments in sensory neuroscience have highlighted the pivotal role of ion channels in mediating ocular surface homeostasis, pain signaling, and inflammation. This review examines the role of the following major ion channel families in the pathophysiology of DED and neuropathic ocular pain: transient receptor potential (TRP) channels, voltage-gated sodium (Nav) channels, and purinergic P2X receptors. The review details their anatomical distribution, molecular function, and responses to environmental stimuli such as heat, cold, osmolarity, and injury. Current treatments, such as artificial tears, anti-inflammatory drops, and systemic neuromodulators, are also reviewed in relation to their effects on ion channel modulation. Additionally, emerging therapies that directly target sensory transduction pathways are introduced. This review highlights the therapeutic potential of ion channel modulation in personalizing treatment for patients with ocular surface pain, particularly those with neuropathic features unresponsive to standard care. Full article

(This article belongs to the Special Issue Latest Review Papers in Molecular Biology 2026)

► Show Figures

Figure 1

27 pages, 16894 KB

Open AccessArticle

MeNADP-ME3 Confers Salt and Drought Tolerance in Arabidopsis and Drives Functional Diversification of the NADP-ME Family in Cassava

by Shuwen Wu, Zhanming Xia, Jiazheng Zhao, Changyi Wang, Yi Min and Dayong Wang

Curr. Issues Mol. Biol. 2026, 48(3), 331; https://doi.org/10.3390/cimb48030331 - 20 Mar 2026

Viewed by 304

Abstract

As a typical C3-C4 intermediate plant, cassava (Manihot esculenta Crantz) exhibits high photosynthetic efficiency and low photorespiration. NADP-malic enzyme (NADP-ME) is a key enzyme in the C4 photosynthetic pathway that provides elevated

{CO}_{2}

concentrations for Rubisco. However, research on NADP-ME in [...] Read more.

As a typical C3-C4 intermediate plant, cassava (Manihot esculenta Crantz) exhibits high photosynthetic efficiency and low photorespiration. NADP-malic enzyme (NADP-ME) is a key enzyme in the C4 photosynthetic pathway that provides elevated

{CO}_{2}

concentrations for Rubisco. However, research on NADP-ME in C3-C4 intermediate species remains limited. In this study, we identified four NADP-ME genes in the cassava genome, with segmental duplication serving as the primary driving force for gene evolution. Cis-acting element analysis indicated potential roles of MeNADP-ME genes in environmental adaptation, stress responses, and growth regulation. Expression profiling using bulk RNA sequencing and single-cell RNA sequencing revealed distinct expression patterns in different tissues and cell subsets. Comparative analysis with Arabidopsis (Arabidopsis thaliana) and maize (Zea mays) NADP-ME families demonstrated that MeNADP-ME3 exhibits bundle sheath cell-specific expression analogous to ZmchlC4NADP-ME in maize. Notably, photosynthetic genes and plasmodesmata (PD)-related genes exhibited high co-expression within mesophyll subcluster 13 and bundle sheath cells, providing molecular evidence for a limited C4 photosynthetic pathway in cassava. Protein–protein interaction predictions implicated MeNADP-ME3 in photosynthetic carbon metabolism and photorespiration regulation. Furthermore, qRT-PCR revealed significant responsiveness of MeNADP-ME3 to various abiotic stresses, and confocal imaging confirmed its chloroplast localization. Functional validation demonstrated that Arabidopsis overexpressing MeNADP-ME3 exhibited 30–120% enhanced antioxidant enzyme activities (SOD, POD, CAT) and 20–32% reduced oxidative damage markers (MDA,

H_{2} O_{2}

) under drought and salt stresses. These findings reveal the evolutionary trajectory of NADP-ME genes in C3-C4 intermediate species and provide genetic resources for developing stress-tolerant cassava cultivars. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Plant Stress Responses and Development)

► Show Figures

Figure 1

10 pages, 223 KB

Open AccessArticle

Personalized Immunotherapy in Osteoarthritis: A Clinical Trial of Autologous Dendritic Cell Immunotherapy in Knee Osteo-Arthritis

by Kurniawan Silalahi, Bhimo Aji Hernowo, Jonny Jonny, Lintang Sagoro, Chrismis Novalinda Ginting and Terawan Agus Putranto

Curr. Issues Mol. Biol. 2026, 48(3), 330; https://doi.org/10.3390/cimb48030330 - 20 Mar 2026

Viewed by 364

Abstract

Background/Objectives: Osteoarthritis (OA) is a chronic inflammatory disease with limited disease-modifying therapies. This study explored a novel immunomodulatory approach using autologous, antigen-pulsed semi-mature dendritic cells (DCs) to modulate the inflammatory milieu in knee OA patients. Methods: In this open-label, quasi-experimental study, [...] Read more.

Background/Objectives: Osteoarthritis (OA) is a chronic inflammatory disease with limited disease-modifying therapies. This study explored a novel immunomodulatory approach using autologous, antigen-pulsed semi-mature dendritic cells (DCs) to modulate the inflammatory milieu in knee OA patients. Methods: In this open-label, quasi-experimental study, 29 subjects received a single subcutaneous injection of autologous DCs. Outcomes assessed at baseline and 4 weeks included the WOMAC index for symptoms and serum levels of IL-6 and TNF-α. Responses were analyzed in the overall cohort and by BMI subgroups. Results: The overall cohort showed a non-significant trend in WOMAC improvement (p = 0.080) and no change in IL-6 (p = 0.785) or TNF-α (p = 0.330). Subgroup analysis revealed differential patterns of response: WOMAC scores improved significantly only in normal-weight patients (p = 0.030), while serum TNF-α decreased significantly only in overweight patients (p = 0.025). IL-6 levels were unchanged across all groups. Conclusions: Autologous antigen-pulsed DC administration was associated with differential responses across BMI subgroups. Symptomatic benefit was observed in normal-weight individuals, while a reduction in systemic TNF-α occurred in overweight patients. These findings suggest that the host metabolic state may modulate the response to DC-based immunotherapy, and therefore warrant validation in a randomized, placebo-controlled trial. Full article

(This article belongs to the Special Issue Molecular Mechanisms and Innovative Therapeutic Approaches in Inflammatory Diseases, Pioneering Precision Medicine Solutions, 2nd Edition)

25 pages, 1432 KB

Open AccessReview

Research Progress on Anticancer Mechanism of Ginsenoside Regulating Tumor Microenvironment

by Tianjia Liu, Wei Li, Da Liu and Baiji Xue

Curr. Issues Mol. Biol. 2026, 48(3), 329; https://doi.org/10.3390/cimb48030329 - 20 Mar 2026

Viewed by 431

Abstract

Cancer is currently one of the most significant health threats facing humanity in general. The clinical treatment of cancer is constrained by the current development of chemotherapy drug resistance, poor pharmacokinetics, off-target toxicity, and insufficient intratumoral accumulation. Although surgery combined with chemotherapy is [...] Read more.

Cancer is currently one of the most significant health threats facing humanity in general. The clinical treatment of cancer is constrained by the current development of chemotherapy drug resistance, poor pharmacokinetics, off-target toxicity, and insufficient intratumoral accumulation. Although surgery combined with chemotherapy is now maturely used in clinical practice, the results are unsatisfactory, and the incidence and mortality of cancer continue to increase year by year with high side effects from treatment. Therefore, it is important to find more effective therapeutic targets against cancer. Alterations in the tumor microenvironment can lead to cellular gene mutations, which are an important cause of tumorigenesis, and therapeutic interventions targeting the tumor microenvironment have been one of the most interesting research areas in the oncology community in recent years. Ginseng is rich in antitumor-active ingredients and is used in the treatment of many cancer diseases. Ginsenoside is one of the main active components of ginseng. This paper reviews the antitumor mechanism of action of ginsenoside through regulating the tumor microenvironment, emphasizing the key role of ginsenoside in the tumor microenvironment and providing a new target and theoretical basis for ginsenoside in the treatment of cancer. Full article

(This article belongs to the Special Issue Advances in Pharmacotherapeutic Strategies to Prevent Tumor Development, Progression and Treatment Resistance, 3rd Edition)

► Show Figures

Figure 1

14 pages, 746 KB

Open AccessArticle

Human Endogenous Retroviruses and Epigenetic Regulators Are Dysregulated in Beckwith–Wiedemann Syndrome

by Ilaria Galliano, Pier-Angelo Tovo, Cristina Calvi, Anna Pau, Anna Clemente, Paola Montanari, Stefano Gambarino, Alessandro Mussa and Massimiliano Bergallo

Curr. Issues Mol. Biol. 2026, 48(3), 328; https://doi.org/10.3390/cimb48030328 - 19 Mar 2026

Viewed by 337

Abstract

Beckwith–Wiedemann syndrome (BWS) is an overgrowth disorder caused by genetic and epigenetic alterations at chromosome 11p15.5. Increasing evidence suggests that imprinting defects may be accompanied by broader epigenomic perturbations affecting repetitive elements such as human endogenous retroviruses (HERVs). We quantified the transcriptional levels [...] Read more.

Beckwith–Wiedemann syndrome (BWS) is an overgrowth disorder caused by genetic and epigenetic alterations at chromosome 11p15.5. Increasing evidence suggests that imprinting defects may be accompanied by broader epigenomic perturbations affecting repetitive elements such as human endogenous retroviruses (HERVs). We quantified the transcriptional levels of the HERV-H, HERV-K, and HERV-W-pol genes, the HERV-derived env genes, Syncytin-1 (SYN1) and Syncytin-2 (SYN2), and the epigenetic regulators, TRIM28 and SETDB1, in whole blood from children and adolescents with BWS, stratified by molecular subtype (ICR2 loss of methylation, n = 14; UPD(11)pat, n = 10), and compared with age-matched healthy controls using quantitative real-time PCR. The BWS samples showed significantly increased transcription of HERV-H and HERV-K relative to controls, whereas HERV-W was unchanged. The SYN1 transcripts were significantly higher in UPD(11)pat compared with controls, while SYN2 did not differ between groups. TRIM28 and SETDB1 were significantly overexpressed in BWS, irrespective of molecular subtype, and no significant differences were observed between ICR2 and UPD(11)pat for HERV-H, HERV-K, HERV-W, TRIM28, or SETDB1. These findings indicate selective dysregulation of endogenous retroelements and key repressors in BWS, consistent with epigenetic alterations extending beyond canonical imprinted loci. Full article

(This article belongs to the Section Molecular Medicine)

► Show Figures

Figure 1

24 pages, 425 KB

Open AccessArticle

Phytochemical Profile and Bioactive Potential of Hampea rovirosae Standl.: Antioxidant, Antimicrobial, and Carbohydrate-Hydrolyzing Enzyme Inhibitory Activities

by Maria Candelaria Tejero-Rivas, José Rodolfo Velázquez-Martínez, Minerva Aurora Hernández-Gallegos, Angelica Alejandra Ochoa-Flores, Rodolfo Osorio-Osorio, Juan Guzmán-Ceferino, Emmanuel Cabañas-García and Josafat Alberto Hernandez-Becerra

Curr. Issues Mol. Biol. 2026, 48(3), 327; https://doi.org/10.3390/cimb48030327 - 19 Mar 2026

Viewed by 333

Abstract

Hampea rovirosae Standl. is traditionally used by local communities to treat infections, pain-related conditions, and to reduce blood sugar levels. In this investigation, we produced aqueous, ethanolic, and hydroethanolic extracts of H. rovirosae and assessed their antioxidant, antibacterial, and antihyperglycemic properties in [...] Read more.

Hampea rovirosae Standl. is traditionally used by local communities to treat infections, pain-related conditions, and to reduce blood sugar levels. In this investigation, we produced aqueous, ethanolic, and hydroethanolic extracts of H. rovirosae and assessed their antioxidant, antibacterial, and antihyperglycemic properties in addition to their phytochemical profiles and contents. The phytochemical characterization was performed through a targeted chromatographic and mass spectrometric analysis of phenolic compounds and the quantitation of total phenolic content (TPC), total flavonoid content (TFC), and total tannin content (TTC) by spectrometric assays. The antioxidant capacity was assessed using the DPPH, ABTS, and FRAP assays, and the antibacterial activity was determined by disk diffusion (DD) and minimum inhibitory concentration (MIC) methods. In addition, antihyperglycemic activity was evaluated by inhibiting α-amylase and α-glucosidase. Phytochemical analysis was performed using high-performance liquid chromatography coupled with mass spectrometry (HPLC-MS), employing a targeted analysis approach based on comparing retention times and fragmentation patterns with standards and databases. This analysis revealed a phytochemical profile dominated by phenolic compounds, with quercetin-3-glucoside (155,930.2), caffeic acid (134,399.1), catechin (98,408.8), procyanidin B2 (85,661.7), protocatechuic acid (83,824.3), and epicatechin (53,704.1) being the major metabolites. The hydroethanolic extract exhibited the highest phenolic (426.70 mg GAE/g), flavonoids (119.17 mg CE/g), and tannin (324.46 mg GAE/g) contents, as well as the strongest antioxidant capacity in the DPPH and FRAP assays. Regarding the antibacterial effects, the aqueous extract inhibited Salmonella typhimurium and Escherichia coli, while the hydroethanolic extract was active against S. aureus, B. cereus, and B. subtilis. In enzyme inhibition assays, the hydroethanolic extract showed strong α-glucosidase inhibition and moderate α-amylase inhibition. The findings provide preliminary scientific evidence of the antioxidant and biological activities of Hampea rovirosae in vitro, supporting its traditional use, which should be validated through vivo trials. Full article

(This article belongs to the Special Issue Pharmacological Activities and Mechanisms of Action of Natural Products, 2nd Edition)

11 pages, 2156 KB

Open AccessArticle

Tempol Attenuates Methotrexate-Induced Osteotoxicity via Antioxidant Mechanisms: Impairment of Protection by GPX4 Inhibition Through ML210

by Osman Fatih Arpağ, Fariz Selimli, Ahmet Can Haskan, Muhammed Said Altun, Soner Mete and Halil Mahir Kaplan

Curr. Issues Mol. Biol. 2026, 48(3), 326; https://doi.org/10.3390/cimb48030326 - 19 Mar 2026

Viewed by 291

Abstract

Purpose: Osteotoxicity is a well-recognized adverse effect of Methotrexate (MTX) therapy, primarily driven by oxidative stress and impaired bone remodeling. This study aimed to investigate the protective effects of Tempol, a membrane-permeable nitroxide antioxidant, against MTX-induced osteotoxicity, and to assess how these effects [...] Read more.

Purpose: Osteotoxicity is a well-recognized adverse effect of Methotrexate (MTX) therapy, primarily driven by oxidative stress and impaired bone remodeling. This study aimed to investigate the protective effects of Tempol, a membrane-permeable nitroxide antioxidant, against MTX-induced osteotoxicity, and to assess how these effects are influenced by ML210, a glutathione peroxidase 4 (GPX4) inhibitor. Methods: Murine osteocyte-like MLO-Y4 cells were treated with MTX alone, Tempol alone, or a combination of MTX with Tempol and ML210. Apoptotic markers (caspase-3, Bax, Bcl-2), MAPK signaling proteins (p-JNK, p-ERK), and oxidative stress parameters (TAS, TOS, SOD, GPx) were measured via ELISA to evaluate the redox and apoptotic responses. Results: MTX significantly induced apoptosis, as evidenced by increased caspase-3 activity and Bax expression, along with decreased Bcl-2 levels. MTX also activated the MAPK pathway by upregulating p-JNK and p-ERK. Furthermore, MTX decreased TAS, SOD, and GPx levels, while increasing TOS. Tempol treatment successfully reversed these effects, restoring apoptotic balance, inhibiting MAPK activation, and enhancing antioxidant capacity. However, co-treatment with ML210 markedly attenuated Tempol’s protective effects, resulting in sustained oxidative stress, elevated apoptotic markers, and persistent MAPK pathway activation. This suggests that Tempol’s cytoprotective actions are dependent on functional GPX4 activity. Conclusion: Tempol exhibits strong potential as an adjunctive antioxidant therapy to counteract MTX-induced osteotoxicity. Nevertheless, its efficacy is significantly influenced by the status of the endogenous antioxidant enzyme GPX4. These findings underscore the need for further investigation into Tempol’s mechanism of action in redox-dependent pathways and its suitability in clinical settings, especially where GPX4 function may be compromised. Full article

(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)

► Show Figures

Figure 1

13 pages, 2316 KB

Open AccessArticle

Changes in the Structure of the Neuromuscular Junction and Muscle Fiber Types Following an Acute Injury Model Induced by Eccentric Contraction

by Mariana Baptista, Jurandyr Pimentel Neto, Matheus Bertanha Fior, Isabella Gomes and Adriano Polican Ciena

Curr. Issues Mol. Biol. 2026, 48(3), 325; https://doi.org/10.3390/cimb48030325 - 19 Mar 2026

Viewed by 408

Abstract

The neuromuscular junction (NMJ) is responsible for transmitting neural signals that trigger muscle contraction. Muscle injuries cause damage to cellular structures and trigger local inflammatory processes. In this context, eccentric contraction was used as an experimental model because it involves excessive stretching, generating [...] Read more.

The neuromuscular junction (NMJ) is responsible for transmitting neural signals that trigger muscle contraction. Muscle injuries cause damage to cellular structures and trigger local inflammatory processes. In this context, eccentric contraction was used as an experimental model because it involves excessive stretching, generating mechanical stress. Twenty-five adult male Wistar rats were distributed into groups: Control (C) (n = 5) and Injury (I) (n = 20). The protocol was performed on a treadmill and consisted of 18 sets/5 min/16 m/min speed, with intervals, and with a negative incline (−16º). The analyses consisted of histochemical techniques, such as myofibrillar ATPase and immunofluorescence (calcium channels, synaptophysin and α-bungarotoxin). Group I-0H showed alterations in the presynaptic region and an increase in Type I fibers. I-24H presented disorganization in the postsynaptic region. In I-4D, we observed the reorganization of neuromuscular activity, while I-7D presented greater density and cross-sectional area (CSA) of Type II fibers. It is concluded that the protocol promotes changes in NMJ structure and fiber distribution, mainly in I-24H. In I-4d, a reorganization of neuromuscular activity is observed, and in I-7D, a structural indicator consistent with recovery demonstrates the skeletal muscle’s ability to adapt to injury. Full article

(This article belongs to the Special Issue Molecular Mechanisms of the Neuro-Musculoskeletal System)

► Show Figures

Figure 1

23 pages, 10592 KB

Open AccessArticle

Evaluation of Antitumor and Antimicrobial Photobiological Activity of Nanocarrier Containing Photosensitizer and Magnetic Nanoparticle

by Raphaela Aparecida Schuenck Rodrigues, Sandro Pinheiro da Costa, Veronica da Silva Cardoso, Alane Beatriz Vermelho, Ralph Santos-Oliveira, Franklin Chimaobi Kenechukwu and Eduardo Ricci-Junior

Curr. Issues Mol. Biol. 2026, 48(3), 324; https://doi.org/10.3390/cimb48030324 - 19 Mar 2026

Viewed by 309

Abstract

Nanotechnology combined with photodynamic therapy (PDT) has been explored to enhance antitumor and antimicrobial photobiological activity. Aluminum phthalocyanine chloride (Al-Pc-Cl), with or without magnetic nanoparticles (MagNPs), was incorporated into polymeric nanoparticles (PNPs) to improve the PDT for treating tumors and infectious diseases. Three [...] Read more.

Nanotechnology combined with photodynamic therapy (PDT) has been explored to enhance antitumor and antimicrobial photobiological activity. Aluminum phthalocyanine chloride (Al-Pc-Cl), with or without magnetic nanoparticles (MagNPs), was incorporated into polymeric nanoparticles (PNPs) to improve the PDT for treating tumors and infectious diseases. Three batches of the nanoparticles (MagNPs, PNPs-PS and PNPs-PS-MagNPs) were developed and characterized in terms of size, PdI, morphology by TEM, release study, and antitumor (against A549 cells) and antimicrobial (against MRSA and C. albicans) photobiological activity. The developed nanoparticles were nanometric in size, with MagNPs, PNPs-PS, and PNPs-PS-MagNPs showing 33.6, 186.9, and 333.5 nm, respectively, maintained the magnetic properties (for MagNPs and PNPs-PS-MagNPs), and provided slow and sustained release of the photosensitizer. PNPs-PS and PNPs-PS-MagNPs showed excellent antitumor photobiological activity with cell viabilities of 42 and 34%, respectively, and were not cytotoxic in the dark, with cell viabilities above 70%. PNPs-PS showed strong antibacterial activity against MRSA with an IC₅₀ of 8.26 μg/mL, which was lower to free Al-Pc-Cl with an IC₅₀ of 14.22 μg/mL after I radiation. The results of the antifungal photobiological activity against C. albicans were excellent, with IC₅₀ values of 3.75 and 3.5 μg/mL for PNPs-PS and PNPs-PS-MagNPs, respectively, values which were significantly lower with p < 0.05 than free PS (IC₅₀ > 30 μg/mL) after irradiation with light and fluconazole (IC₅₀ > 30 μg/mL), the reference antifungal agent. PNPs-PS showed promising results regarding antitumor, antibacterial, and antifungal photobiological activity. However, PNPs-PS-MagNPs showed weak results for antibacterial photobiological activity against MRSA but with promising results for tumor cells and C. albicans. Full article

(This article belongs to the Special Issue Emerging Trends in Nanobiotechnology and Nanomedicine)

► Show Figures

Figure 1

22 pages, 8786 KB

Open AccessArticle

PSMB8 as a Core Target Mediating the Anti-Hepatocellular Carci-Noma Activity of Lingonberry (Vaccinium vitis-idaea L.) Extract in HepG2 Cells

by Liangyu Zhu, Zhi Zhang, Yandong Zhang, Dianwen Wei, Zhenyu Wang and Liping Zhou

Curr. Issues Mol. Biol. 2026, 48(3), 323; https://doi.org/10.3390/cimb48030323 - 18 Mar 2026

Viewed by 282

Abstract

Hepatocellular carcinoma (HCC) is a highly malignant tumour with a poor prognosis and few effective treatment options. Development of resistance to conventional therapies and occurrence of severe side effects highlight the urgent need for novel, low-toxicity interventions. Natural products are promising candidates for [...] Read more.

Hepatocellular carcinoma (HCC) is a highly malignant tumour with a poor prognosis and few effective treatment options. Development of resistance to conventional therapies and occurrence of severe side effects highlight the urgent need for novel, low-toxicity interventions. Natural products are promising candidates for HCC drug development thanks to their multi-target activity and favourable safety profiles. Previous studies reported that Lingonberry extract, a bioactive natural product, inhibits proliferation of HepG₂ cells. However, the key molecular targets and underlying anticancer mechanisms remain unclear. In this study, we analysed gene chip data from Lingonberry extract-treated HepG₂ tumour-bearing mice using bioinformatics tools, employing a cross-species, multi-level screening strategy to identify PSMB8 as the core regulatory gene. In vitro functional validations (Western blotting, RT-PCR, CCK-8 assay, colony formation assay, flow cytometry and TUNEL staining) confirmed these findings. Downregulating PSMB8 was found to effectively induce late apoptosis in HepG₂ cells, and Lingonberry extract was found to significantly reduce PSMB8 protein expression. This study identifies PSMB8 as a key mediator of the anticancer effect of Lingonberry extract in HepG₂ cells. It provides a reliable methodological reference for screening anticancer targets of natural products and supports further exploration of Lingonberry extract as a potential adjuvant/lead compound for HCC. Full article

(This article belongs to the Special Issue Pharmacological Activities and Mechanisms of Action of Natural Products, 2nd Edition)

► Show Figures

Graphical abstract

16 pages, 3953 KB

Open AccessArticle

PDGFD: A Dual-Function Regulator That Maintains Myoblast Pool and Fuels Myogenic Differentiation

by Hongzhen Cao, Jing Wang, Yunzhou Wang, Jingsen Huang, Wei Chen, Hui Tang, Junfeng Chen, Baosong Xing and Yongqing Zeng

Curr. Issues Mol. Biol. 2026, 48(3), 322; https://doi.org/10.3390/cimb48030322 - 18 Mar 2026

Viewed by 266

Abstract

The role of platelet-derived growth factor D (PDGFD) in mesenchymal cells is well-established, but its specific function in skeletal muscle generation remains unknown. This study reveals for the first time PDGFD’s dual regulatory role in myogenesis: it acts both as a [...] Read more.

The role of platelet-derived growth factor D (PDGFD) in mesenchymal cells is well-established, but its specific function in skeletal muscle generation remains unknown. This study reveals for the first time PDGFD’s dual regulatory role in myogenesis: it acts both as a “guardian” maintaining the myoblast pool and as an “initiator” driving myogenic differentiation. Through single-cell RNA sequencing analysis of skeletal muscle from Jiangquan Black pigs, we identified PDGFD as a common candidate gene for both muscle and fat development. In the C2C12 cell model, PDGFD knockdown significantly inhibited cell proliferation and promoted apoptosis, while overexpression enhanced viability and inhibited apoptosis, indicating its critical role in maintaining myoprogenic precursor cell homeostasis. Further studies revealed that PDGFD interference downregulated key myogenic differentiation markers MyoD and MyoG, inhibiting differentiation. Its expression peaked during mid-differentiation (D5), suggesting temporal regulation of differentiation. Interestingly, although PDGFD primarily acts through the PI3K/Akt pathway downstream of PDGFR-β, PDGFD knockdown did not show significant synergistic effects with PI3K/Akt pathway activation in inhibiting differentiation. This suggests PDGFD may specifically regulate myogenic differentiation via an independent or parallel signaling axis. This study not only expands the known functions of PDGFD in muscle biology but also provides new insights into the mechanisms by which growth factors coordinate cell fate decisions. Full article

(This article belongs to the Special Issue Molecular and Cellular Insights into the Musculoskeletal System, Joints and Spine)

► Show Figures

Figure 1

12 pages, 5258 KB

Open AccessArticle

Comparative Transcriptomic Analysis Reveals Salt Stress Adaptation Mechanisms in Cultivated Rice Varieties (Oryza sativa)

by Zihao Yuan, Ziqi Liu, Shengyu Mo, Feng Wang, Wuge Liu, Dilin Liu, Wu Yang, Yilong Liao, Leiqing Chen, Le Kong, Hui Wang, Tao Guo and Xing Huo

Curr. Issues Mol. Biol. 2026, 48(3), 321; https://doi.org/10.3390/cimb48030321 - 18 Mar 2026

Viewed by 357

Abstract

Salt stress is an injurious concern of global climate change that negatively impacts the growth and yield of rice plants. Identifying salt tolerance genes is essential to understanding the molecular mechanism regulating salt tolerance in rice. In this study, we treated two rice [...] Read more.

Salt stress is an injurious concern of global climate change that negatively impacts the growth and yield of rice plants. Identifying salt tolerance genes is essential to understanding the molecular mechanism regulating salt tolerance in rice. In this study, we treated two rice varieties, Xiangxiuzhan (XXZ) and Changxiang (CXG), with 100 mM NaCl to examine the effect on the germination and growth stages. Transcriptome analysis was investigated for changes in gene expression between the two varieties. During the germination stage, the CXG variety had higher germination potential than the XXZ variety, whereas in the growth stage, the XXZ variety showed higher survival efficiency than the CXG variety. Transcriptome analysis showed that the XXZ variety had more DEGs in grains, while CXG displayed greater DEGs in leaves and roots. Gene Ontology (GO) and KEGG pathway showed that beta-alanine metabolism, cutin biosynthesis, and plant hormone signal transduction were over-represented, whereas heatmap analysis showed cellular and environmental signal transduction. This study focuses on the molecular pathways of the salt stress tolerance mechanism of Xiangxiuzhan and Changxiang varieties. Full article

(This article belongs to the Special Issue Plant Hormones, Development, and Stress Tolerance)

► Show Figures

Figure 1

19 pages, 1883 KB

Open AccessArticle

Effects of Hybridization and Triploidization on Transcription of Core Metabolic and Stress Response Genes in Rainbow Trout (Oncorhynchus mykiss) × Brook Trout (Salvelinus fontinalis) Hybrids—Preliminary Results

by Marcin Kuciński, Rafał Rożyński and Konrad Ocalewicz

Curr. Issues Mol. Biol. 2026, 48(3), 320; https://doi.org/10.3390/cimb48030320 - 17 Mar 2026

Viewed by 375

Abstract

The transcriptomic effects of hybridization and triploidization were investigated in diploid and triploid rainbow trout, diploid brook trout, as well as triploid hybrids of rainbow trout and brook trout. The examined fish were reared under identical conditions for about two and a half [...] Read more.

The transcriptomic effects of hybridization and triploidization were investigated in diploid and triploid rainbow trout, diploid brook trout, as well as triploid hybrids of rainbow trout and brook trout. The examined fish were reared under identical conditions for about two and a half years after hatching. Expression of ten genes involved in cellular respiration (Atp5bp, Slc25a5), mitochondrial functioning (Mrpl28, Micu2), ribosome biogenesis (Rpl24, Rps24), proteasome-mediated protein turnover (Derl1, Psmc2), and protein chaperoning (Hsp90B1, Pdia4) was studied in liver and muscle tissues. Most of the analyzed genes (Atp5bp, Slc25a5, Mrpl28, Micu2, Rpl24, Rps24, Derl1, and Psmc2) displayed comparable expression levels in the liver tissue across the examined triploid hybrids and diploid parental species, with stabilization of genes that were both positively and negatively compensated in the triploid rainbow trout. In turn, significant upregulation of Slc25a5, Derl1, Rps24, and Rpl24 genes, together with downregulation of Micu2 gene, was observed in the triploid rainbow trout liver and muscle, respectively. On the other hand, triploid hybrids showed marked transcriptional upregulation of genes primarily associated with energy metabolism and protein synthesis (Atp5pb, Slc25a5, Rpl24, Rps24, and Pdia4) relative to all the fish groups examined. Although protein-synthesis- and energy-related genes were upregulated in the muscles of triploid hybrids, the recorded growth performance data did not indicate clear evidence of growth heterosis (MPH = −14.3% for body weight; MPH = −0.4% for body length), suggesting that potential benefits of increased heterozygosity in this cross may not be fully reflected in enhanced growth. Three- to four-fold downregulation of the heat shock protein (Hsp90B1) gene was also observed in both tissues of triploid hybrids compared with purebred diploid and triploid trout, which may reflect potential maladaptive genomic effects commonly observed in distant salmonid crosses, suggesting altered stress-response regulation in the examined triploid hybrids. Full article

(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)

► Show Figures

Figure 1

19 pages, 947 KB

Open AccessArticle

Ultrasound-Assisted Synthesis and Biological Profiling of 1,3,5-Triazine Derivatives with Antiproliferative Activity in Triple-Negative Breast Cancer

by Natalia Bosak, Anna Karolina Drabczyk, Jolanta Jaśkowska, Martyna Stachowicz-Suhs, Beata Filip-Psurska, Anna Boguszewska-Czubara, Katarzyna Ewa Greber, Krzesimir Ciura and Damian Kułaga

Curr. Issues Mol. Biol. 2026, 48(3), 319; https://doi.org/10.3390/cimb48030319 - 17 Mar 2026

Viewed by 315

Abstract

Triple-negative breast cancer (TNBC) remains one of the most aggressive breast cancer subtypes and is associated with limited therapeutic options, underscoring the urgent need for novel treatment strategies. In this study, a library of seventeen 1,3,5-triazine derivatives potentially targeting TNBC was developed using [...] Read more.

Triple-negative breast cancer (TNBC) remains one of the most aggressive breast cancer subtypes and is associated with limited therapeutic options, underscoring the urgent need for novel treatment strategies. In this study, a library of seventeen 1,3,5-triazine derivatives potentially targeting TNBC was developed using an activity-based approach. Compounds were synthesized via an ultrasound-assisted protocol, providing an efficient and environmentally friendly methodology. The synthesized library was evaluated in vitro against the human TNBC cell lines MDA-MB-468, MDA-MB-231, and Hs578T, as well as the non-tumorigenic epithelial cell line MCF10A. Compounds 9 and 17 exhibited the most promising antiproliferative activity against TNBC cell lines (MDA-MB-468: IC₅₀ = 36.62 µM for 9 and 38.29 µM for 17; MDA-MB-231: IC₅₀ = 37.32 µM for 9 and 32.86 µM for 17; Hs578T: IC₅₀ = 57.26 µM for 9 and 34.87 µM for 17), while maintaining acceptable selectivity toward non-cancerous cells. The lead compounds were further assessed in vivo using a Danio rerio model to evaluate general toxicity and cardiotoxicity. In addition, ADME parameters were predicted for all compounds using biomimetic chromatography. Overall, compounds 9 and 17 emerged as promising small-molecule candidates for TNBC treatment, requiring further toxicological evaluation in more human-relevant in vivo models. Full article

(This article belongs to the Special Issue Targeted Therapeutic Approaches in Cancer: Combining Natural Compounds and Conventional Drugs)

► Show Figures

Figure 1

13 pages, 1601 KB

Open AccessArticle

Exploring the Antimicrobial Potential of a Novel Phage-Derived Lytic Protein Against Pseudomonas aeruginosa

by Sibongile Mtimka, Kanyane Bridgett Malatji, Patrick Opare Sakyi, Noel David Nogbou, Andrew Munyalo Musyoki, Sipho Mamputha, Lusisizwe Kwezi, Samuel Kojo Kwofie, Ofentse Jacob Pooe and Tsepo Lebiletsa Tsekoa

Curr. Issues Mol. Biol. 2026, 48(3), 318; https://doi.org/10.3390/cimb48030318 - 17 Mar 2026

Viewed by 335

Abstract

The escalation of bacterial resistance to existing antibiotics represents a growing global health challenge, exacerbated by the widespread misuse of antimicrobial agents. As a result, alternative antibacterial strategies are increasingly being explored, including phage-derived lytic proteins. In this study, we report a preliminary [...] Read more.

The escalation of bacterial resistance to existing antibiotics represents a growing global health challenge, exacerbated by the widespread misuse of antimicrobial agents. As a result, alternative antibacterial strategies are increasingly being explored, including phage-derived lytic proteins. In this study, we report a preliminary characterisation of a novel phage-derived lytic protein identified through computational screening of bacteriophage genome sequences. A putative open reading frame, designated SM07 (1383 bp), was selected from bacteriophage sequences contributed by the University of KwaZulu-Natal to a global phage repository. The gene was synthesised, sub-cloned into the pET-30b(+) vector with an N-terminal histidine tag, and recombinantly expressed in Escherichia coli BL-21(AI) cells. The protein was purified using affinity and ion-exchange chromatography. Purified SM07 exhibited in vitro antimicrobial activity against Pseudomonas aeruginosa, with a minimum inhibitory concentration of 4 µg/mL, while no significant cytotoxic effects were observed in Vero kidney cells at concentrations substantially above the effective dose. Together, these findings provide initial evidence supporting the antimicrobial potential of SM07 and highlight phage-derived lytic proteins as candidates for further investigation as alternative agents against P. aeruginosa-associated infections. Full article

(This article belongs to the Section Molecular Microbiology)

► Show Figures

Figure 1

17 pages, 2757 KB

Open AccessArticle

Time-Series-Based Co-Expression Network Analysis Reveals Key Regulatory Modules and Hub Genes in Salt-Tolerant Wheat Under Salt Stress

by Guiqiang Fan, Jianan Huang, Hong-Jin Wang, Yuxiang Huo, Peiyu Liu, Uzair Ullah, Guohang Hu, Munib Ahmad, Abdullah Shalmani, Hui Fang and Tianrong Huang

Curr. Issues Mol. Biol. 2026, 48(3), 317; https://doi.org/10.3390/cimb48030317 - 16 Mar 2026

Viewed by 353

Abstract

Salt stress severely constrains wheat growth and yield by inducing osmotic imbalance, ion toxicity, and excessive accumulation of reactive oxygen species (ROS). Although salt-tolerant cultivars can adapt through rapid signaling transduction and maintenance of cellular homeostasis, the underlying dynamic regulatory networks remain insufficiently [...] Read more.

Salt stress severely constrains wheat growth and yield by inducing osmotic imbalance, ion toxicity, and excessive accumulation of reactive oxygen species (ROS). Although salt-tolerant cultivars can adapt through rapid signaling transduction and maintenance of cellular homeostasis, the underlying dynamic regulatory networks remain insufficiently characterized. In this study, we reanalyzed publicly available time-series RNA-seq data (0, 1, 3, 6, 12, and 24 h) from the salt-tolerant wheat cultivar Xiaoyan22 under salt stress and constructed a time-series-based co-expression network using weighted gene co-expression network analysis (WGCNA). Multiple gene modules were identified, among which the black module showed significant positive correlations with both salt treatment (treatment_bin) and stress duration (time_h). This module displayed a progressively increasing eigengene expression pattern throughout the stress period. Gene significance (GS) was positively correlated with module membership (MM), facilitating the identification of highly connected hub genes within this module. Functional enrichment analysis indicated that genes in the black module were primarily associated with DNA replication and genome stability maintenance, RNA metabolic regulation, phenylpropanoid metabolism, and cuticle/suberin/wax biosynthesis. Physiological analysis further revealed enhanced activities of superoxide (SOD), peroxide (POD), and catalase (CAT), enhanced accumulation of proline and soluble sugars, and a time-dependent increase in MDA under salt stress. qRT-PCR confirmed significant induction of candidate genes, including a ZAR1-like receptor kinase, Remorin, and NETWORKED 1D. Collectively, these findings integrate co-expression network inference with physiological and molecular validation, providing candidate regulators and pathways for understanding salt tolerance and supporting future molecular breeding efforts. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Plant Adaptation and Stress Tolerance Under Changing Environmental Conditions)

► Show Figures

Figure 1

16 pages, 5220 KB

Open AccessArticle

Dual Inhibition of GSK3 and JAK by BIO Suppresses Osteoblast Differentiation and Mineralization of Human Mesenchymal Cells

by Nihal Almuraikhi, Latifa Alkhamees, Sumaiya Tareen and Manikandan Muthurangan

Curr. Issues Mol. Biol. 2026, 48(3), 316; https://doi.org/10.3390/cimb48030316 - 16 Mar 2026

Viewed by 308

Abstract

Glycogen synthase kinase-3 (GSK3) inhibition is a commonly used approach to promote osteogenic differentiation through activation of Wnt signaling. However, 6-bromoindirubin-3′-oxime (BIO), which is commonly used for GSK3 inhibition, also targets JAK/STAT, raising the possibility of dual pathway interference during osteoblast differentiation, as [...] Read more.

Glycogen synthase kinase-3 (GSK3) inhibition is a commonly used approach to promote osteogenic differentiation through activation of Wnt signaling. However, 6-bromoindirubin-3′-oxime (BIO), which is commonly used for GSK3 inhibition, also targets JAK/STAT, raising the possibility of dual pathway interference during osteoblast differentiation, as both GSK3 and JAK/STAT pathways are critical regulators of osteoblastogenesis. In this study, we investigated the effect of BIO on the osteoblast differentiation of hMSCs-TERT4. While BIO had no significant effect on cell viability or apoptosis, it markedly inhibited osteoblast differentiation, as evidenced by reduced ALP activity, decreased matrix mineralization, and downregulation of osteoblast-associated markers. Microarray analysis followed by qRT-PCR validation revealed downregulation of Wnt and TGF-β pathway genes. These findings show that BIO suppresses osteoblast commitment and osteogenic differentiation, accompanied by altered Wnt- and TGF-β-related gene expression. This study provides mechanistic insight into the off-target consequences of widely used small molecules and highlights the importance of dissecting pathway-specific roles in stem cell differentiation. Understanding the interplay between GSK3 and JAK signaling is essential for optimizing pharmacological strategies in skeletal regenerative medicine. This study highlights the importance of pathway selectivity when using small molecules in stem cell-based therapies for bone regeneration. Full article

(This article belongs to the Special Issue Molecular Mechanisms and Therapeutic Targets in Bone Biology and Cancer)

► Show Figures

Figure 1

13 pages, 2942 KB

Open AccessArticle

American Ginseng (Panax quinquefolius) Extracts (G1899) Ameliorate Immunosenescence via Regulation of T Cell Populations and Aging-Related Proteins in a Mouse Model Induced by D-Galactose and Tert-Butyl Hydroperoxide

by Ji-Hye Park, Jaehoon Lee, Chang Hwan Lee, Sun Hee Hyun and Seung-Ho Lee

Curr. Issues Mol. Biol. 2026, 48(3), 315; https://doi.org/10.3390/cimb48030315 - 16 Mar 2026

Viewed by 389

Abstract

Immunosenescence is characterized by an age-associated decline in immune function, particularly involving T-cell dysfunction, which increases susceptibility to infections and chronic diseases. This study investigated the anti-aging and immunomodulatory effects of American ginseng extract (G1899) using in vitro and in vivo models of [...] Read more.

Immunosenescence is characterized by an age-associated decline in immune function, particularly involving T-cell dysfunction, which increases susceptibility to infections and chronic diseases. This study investigated the anti-aging and immunomodulatory effects of American ginseng extract (G1899) using in vitro and in vivo models of aging. Cellular senescence was induced in HepG2 cells by D-galactose treatment, followed by exposure to G1899 (20 and 100 μg/mL). Senescence-associated markers were assessed to evaluate cellular aging. An aging mouse model was established in male C57BL/6 mice through intraperitoneal administration of D-galactose (500 mg/kg) and tert-butyl hydroperoxide (0.4 mmol/kg), and G1899 was orally administered at 400 mg/kg. Thymic immune cell subsets and aging-related protein expression were analyzed using flow cytometry and Western blotting. G1899 significantly reduced p21 expression and senescence-associated β-galactosidase activity in senescent HepG2 cells. In aging-induced mice, G1899 restored CD4⁺ and CD8⁺ T-cell populations, normalized naïve T-cell levels, and reduced anergic CD28-negative T cells. Furthermore, G1899 regulated the expression of key aging-related proteins, including FOXO1, Sirt1, p53, and CD38. These findings demonstrate that G1899 attenuates age-related immune alterations by restoring thymic T-cell homeostasis and regulating aging-associated molecular pathways. Full article

(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)

► Show Figures

Figure 1

20 pages, 1827 KB

Open AccessEditor’s ChoiceArticle

Effects of Citicoline-Based Supplementation on Lipid Peroxidation Markers and Sirtuin-1 Expression in Ischemic Stroke

by Todorka Sokrateva, Bogdan Roussev, Daniela V. Vankova, Deyana G. Vankova, Diana Ivanova, Mihael Tsalta-Mladenov, Darina Georgieva, Miglena N. Nikolova, Galya Mihaylova and Milka A. Nashar

Curr. Issues Mol. Biol. 2026, 48(3), 314; https://doi.org/10.3390/cimb48030314 - 15 Mar 2026

Viewed by 610

Abstract

Ischemic stroke (IS) is associated with pronounced oxidative stress and lipid peroxidation, which contribute to secondary neuronal damage. This study explored the effects of a six-month intervention with a new formulation containing citicoline, vitamin C, and extracts from green tea and aronia (Cytodeox™) [...] Read more.

Ischemic stroke (IS) is associated with pronounced oxidative stress and lipid peroxidation, which contribute to secondary neuronal damage. This study explored the effects of a six-month intervention with a new formulation containing citicoline, vitamin C, and extracts from green tea and aronia (Cytodeox™) on arachidonic acid (AA) metabolism, lipid peroxidation assessed by total 8-iso-prostaglandin F₂α (8-iso-PGF₂α), and Sirtuin-1 (SIRT1) expression in healthy controls (n = 43) and patients with IS (n = 53), both with and without comorbidities. AA and 8-iso-PGF₂α were quantified in serum using UPLC–MS and ELISA, respectively, and the fold change in SIRT1 expression was assessed in peripheral blood mononuclear cells (PBMCs) by RT-qPCR. In healthy controls, Cytodeox™ significantly lowered AA and 8-iso-PGF₂α levels. IS patients showed markedly increased baseline 8-iso-PGF₂α, indicating severe oxidative stress. Following supplementation, 8-iso-PGF₂α levels increased in patients with comorbidities, particularly diabetes mellitus (DM), whereas an exploratory analysis suggested a decreasing trend in patients without comorbidities. SIRT1 expression was significantly upregulated in IS patients, with the most pronounced increase observed in the DM subgroup, while remaining unchanged in controls. These findings suggest a protective, antioxidant, and membrane stabilising effect of Cytodeox™ under conditions of preserved or moderately impaired redox homeostasis, supporting its potential role as a preventive or early supportive intervention. Full article

(This article belongs to the Special Issue Repurposing and Innovation: Drug Research in Neuroprotection)

► Show Figures

Figure 1

17 pages, 2177 KB

Open AccessArticle

Dietary Fructose and Palmitic Acid Induce Shared and Divergent Transcriptional Responses in the Larval Midgut of Drosophila melanogaster

by Laura Castañeda-Partida, Myriam Campos-Aguilar, Luis Felipe Santos-Cruz, Lizbeth Abigail Piña-Soto, Santiago Cristobal Sigrist Flores, María Eugenia Heres-Pulido, Irma Elena Dueñas-García, Elías Piedra-Ibarra, Rafael Jiménez-Flores and Alberto Ponciano-Gómez

Curr. Issues Mol. Biol. 2026, 48(3), 313; https://doi.org/10.3390/cimb48030313 - 14 Mar 2026

Viewed by 266

Abstract

Background: High-energy diets enriched in simple sugars and saturated fatty acids alter metabolic homeostasis, yet how distinct nutrients are integrated at the transcriptional level remains incompletely understood. Methods: Here, we profiled the larval midgut transcriptome of Drosophila melanogaster following 24 h exposure to [...] Read more.

Background: High-energy diets enriched in simple sugars and saturated fatty acids alter metabolic homeostasis, yet how distinct nutrients are integrated at the transcriptional level remains incompletely understood. Methods: Here, we profiled the larval midgut transcriptome of Drosophila melanogaster following 24 h exposure to diets enriched with 5% fructose (FD), 1% palmitic acid (PD), or their combination (MD). RNA sequencing (Illumina NovaSeq) was performed on pooled third-instar larval midguts, and differential expression analyses were conducted to identify diet-associated transcriptional changes. Results: The results revealed extensive transcriptional remodeling, with most responses being diet-specific, alongside a conserved core of genes regulated across all treatments. Shared transcriptional signatures were associated with proteostasis and amino acid transport pathways. Comparative and pattern-based analyses further uncovered discordant gene sets and pathway enrichments that were unique to individual diets or to the combined exposure. Notably, the mixed diet induced distinct expression patterns with specific functional signatures that were not predictable from either nutrient alone. Conclusions: Together, these findings indicate that the larval midgut integrates carbohydrate and lipid inputs through coordinated and context-dependent transcriptional responses, highlighting the importance of nutrient combinations in shaping epithelial metabolic programs. Full article

(This article belongs to the Special Issue Molecular Regulation of Insect Adaptation)

► Show Figures

Figure 1

15 pages, 3974 KB

Open AccessArticle

Genome-Wide Identification of SWEET Gene Family and Integrative Transcriptomic–Metabolomic Analysis Reveal Sugar Transport-Mediated Chilling Responses in Sesame (Sesamum indicum L.)

by Pan Zeng, Yunyan Zhao, Junchao Liang, Xiaowen Yan, Zhiqi Wang and Jian Sun

Curr. Issues Mol. Biol. 2026, 48(3), 312; https://doi.org/10.3390/cimb48030312 - 14 Mar 2026

Viewed by 296

Abstract

Sesame is a thermophilic oilseed crop that is vulnerable to low-temperature stress. SWEET sugar transporters are important for sugar allocation, but their roles in sesame cold responses remain poorly understood. In this study, 24 SWEET genes were identified in the sesame genome and [...] Read more.

Sesame is a thermophilic oilseed crop that is vulnerable to low-temperature stress. SWEET sugar transporters are important for sugar allocation, but their roles in sesame cold responses remain poorly understood. In this study, 24 SWEET genes were identified in the sesame genome and classified into six conserved groups with high structural conservation and limited duplication. Comparative transcriptomic and metabolomic analyses of cold-tolerant and cold-sensitive sesame accessions under chilling stress revealed distinct SiSWEET expression patterns and contrasting soluble sugar accumulation. Several SiSWEET genes showed significant correlations with glucose, fructose, and sucrose contents. These results suggest that SWEET-mediated sugar transport is involved in sesame chilling responses and provide candidate genes for improving cold tolerance. Full article

(This article belongs to the Special Issue Molecular Mechanisms of Plant Stress Responses and Development)

► Show Figures

Figure 1

13 pages, 4071 KB

Open AccessArticle

Maresin-1 Alleviates Sepsis-Induced Liver Injury by Regulating Apoptosis and Autophagy via Activation of the PI3K/Akt Signaling Pathway in Mice

by He Wang, Min Sun and Heng Fan

Curr. Issues Mol. Biol. 2026, 48(3), 311; https://doi.org/10.3390/cimb48030311 - 13 Mar 2026

Viewed by 297

Abstract

Sepsis-induced liver injury (SILI) stands as an independent prognostic factor for mortality among patients diagnosed with sepsis. Maresin-1 (MaR1) is a proresolving lipid mediator. However, its significance in SILI is uncertain. The current study sought to investigate MaR1’s effectiveness in treating SILI, as [...] Read more.

Sepsis-induced liver injury (SILI) stands as an independent prognostic factor for mortality among patients diagnosed with sepsis. Maresin-1 (MaR1) is a proresolving lipid mediator. However, its significance in SILI is uncertain. The current study sought to investigate MaR1’s effectiveness in treating SILI, as well as its molecular mechanism. In male C57BL/6J mice, we generated a SILI model by using cecal ligation and puncture (CLP). We further investigated how MaR1 influences inflammation, hepatic autophagy and apoptosis. We showed that treatment with MaR1 ameliorates SILI-induced hepatic injury, as reflected in decreased blood level of the alanine aminotransferase (ALT) and aspartate aminotransferase (AST) enzymes, as well as better appearance of liver tissues. Furthermore, this medication markedly reduced the expression of inflammatory mediators. Importantly, MaR1 inhibited hepatocyte apoptosis by regulating the Bax/Bcl-2 ratio, decreasing cleaved caspase-3 expression, lowering apoptotic cell count, and increasing autophagy. The findings demonstrated that MaR1 treatment reduced p62 protein expression while raising Beclin1 levels and the LC3-II/LC3-I ratio, and facilitated autophagosome formation. The observed effects were most likely due to the stimulation of PI3K/Akt signaling, which was completely prevented by LY294002 (LY), a specific PI3K inhibitor. MaR1’s protective effect in SILI may be mediated via stimulation of the PI3K/Akt pathway, which reduces inflammation and regulates apoptosis and autophagy. Our results give additional experimental evidence of the potential therapeutic uses of MaR1 in the treatment of SILI. Full article

(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)

► Show Figures

Figure 1

15 pages, 6216 KB

Open AccessArticle

Multi-Strain Probiotic and Bee Pollen Supplementation Attenuates CCl₄-Induced Altered Intestinal Tight Junctions in Rodents

by Nada Alsayari, Ramesa Shafi Bhat, Seema Zargar, Abeer M. Aldbass and Sooad Al-Daihan

Curr. Issues Mol. Biol. 2026, 48(3), 310; https://doi.org/10.3390/cimb48030310 - 13 Mar 2026

Viewed by 293

Abstract

Environmental toxins can impair gut microbiota and increase intestinal permeability, contributing to various health problems. While many such toxins are known to disrupt tight junctions and compromise barrier function, research specifically examining carbon tetrachloride (CCl₄) as a trigger of intestinal epithelial [...] Read more.

Environmental toxins can impair gut microbiota and increase intestinal permeability, contributing to various health problems. While many such toxins are known to disrupt tight junctions and compromise barrier function, research specifically examining carbon tetrachloride (CCl₄) as a trigger of intestinal epithelial barrier dysfunction remains limited. In this study, 54 young Western albino male rats, weighing 180–200 g, were randomly assigned to nine experimental groups, each comprising six rats. Group 1 received 1 mL of oral saline and served as a control. Groups 2 and 3 received 0.2 g/kg body weight probiotic and prebiotic, respectively, for four weeks. CCl₄ (1 mL/kg, i.p.) was administered either at the beginning of day 1 (damage induction; Group 4) or at the end of day 28 (protection assessment; Group 7). Intervention groups received probiotics and prebiotics for 4 weeks after (therapeutic) CCl₄ exposure on day 1 in Groups 5 and 6, respectively. Groups 8 and 9 received probiotics and prebiotics for 4 weeks before CCl₄ exposure on day 28, respectively. Quantification of gut bacterial populations, serum levels of Occludin and Zonulin, as biomarkers of intestinal permeability, and histopathological analysis of intestinal tissue were conducted. CCl₄ induces significant intestinal epithelial barrier dysfunction with marked histopathological alterations. Probiotic treatment was more effective than prebiotics at normalizing serum Zonulin and Occludin levels in CCl₄-induced intestinal damage. Probiotics restore microbial balance by suppressing the overgrowth of pathogenic organisms, while prebiotics confer partial protection. CCl₄-induced gut barrier disruption is restored through probiotic supplements by restoring gut microbial balance and normalizing tight junction-associated biomarkers. Full article

(This article belongs to the Section Molecular Microbiology)

► Show Figures

Figure 1

25 pages, 15185 KB

Open AccessArticle

Integrated Metabolomics and Transcriptomics Analysis Reveals the Biosynthetic Mechanism of Isoquinoline Alkaloids in Different Tissues of Hypecoum erectum L.

by Sainan Wang, Yan Du and Meiqing Yang

Curr. Issues Mol. Biol. 2026, 48(3), 309; https://doi.org/10.3390/cimb48030309 - 13 Mar 2026

Viewed by 332

Abstract

Hypecoum erectum L. is a medicinal plant known for its high content of isoquinoline alkaloids (IQAs), a class of compounds with diverse pharmacological activities. To elucidate the biosynthetic mechanisms and tissue-specific accumulation of IQAs, we integrated HPLC-MS/MS-based metabolomic analysis with RNA sequencing (RNA-seq) [...] Read more.

Hypecoum erectum L. is a medicinal plant known for its high content of isoquinoline alkaloids (IQAs), a class of compounds with diverse pharmacological activities. To elucidate the biosynthetic mechanisms and tissue-specific accumulation of IQAs, we integrated HPLC-MS/MS-based metabolomic analysis with RNA sequencing (RNA-seq) transcriptomic profiling across the roots, stems, and leaves of H. erectum. Metabolomic analysis identified twenty-six IQAs as differentially accumulated metabolites (DAMs) among the three tissues, while transcriptomic analysis revealed twenty-two categories of differentially expressed genes (DEGs) involved in IQA biosynthesis. KEGG pathway enrichment analysis demonstrated that nine DAMs and twenty categories of DEGs were co-enriched in the IQA biosynthetic pathway of Hypecoum erectum. Notably, seven key DAMs—Stylopine, Protopine, Magnoflorine, Corydaline, Tetrahydropalmatine, Sanguinarine, and Palmatine—preferentially accumulated in the root, concomitant with the elevated expression of eleven root-specific DEGs, including GOT1, CYP719A14, SMT, CYP719A1_2_3_13, PSOMT1, E2.1.1.116, CYP80B1, E2.1.1.128, NCS, ASP5, and BBE1. Gene–metabolite correlation network analysis further identified nine DAMs and fifteen DEGs closely associated with IQA biosynthesis, highlighting key enzymatic regulators of alkaloid accumulation. Additionally, several transcription factor (TF) families, including bHLH, NAC, and ERF families, were predicted to participate in the transcriptional regulation of IQA-related genes. Collectively, these findings demonstrate that roots are the primary site of IQA biosynthesis in H. erectum and provide a molecular framework for understanding the regulation and utilization of its medicinally active components. Full article

(This article belongs to the Special Issue Advances in Multi-Omics for Functional Genomics Studies and Molecular Breeding, 2nd Edition)

► Show Figures

Figure 1

Journal Menu

Journal Browser

Curr. Issues Mol. Biol., Volume 48, Issue 3 (March 2026) – 100 articles

Further Information

Guidelines

MDPI Initiatives

Follow MDPI