Search Results (1,334)

The proliferation of methicillin-resistant Staphylococcus aureus (MRSA) in food processing is an escalating public health issue. This circumstance has intensified the quest for ecological alternatives to impede pathogen proliferation and avert food degradation. This study firstly investigated the chemical compositions of three essential oils (EOs) sourced from Eucalyptus, Rosemary and Lavender plants using GC-MS. Subsequently, the antibacterial and antibiofilm activities of the tested EOs were assessed against MRSA strains. The effects of these EOs on the expression of antibiotic resistance-related (mecA), virulence regulatory (agrA and sarA), and enterotoxin (sea) genes in MRSA strains were also evaluated by real-time PCR. Concerning the composition analyses performed on the EOs, our results revealed a total of 82 compounds, which accounted for 99.20, 98.10 and 92.78% of Eucalyptus, Rosemary and Lavender EOs, respectively. The anti-staphylococcal activity showed that Eucalyptus EO had the greatest effect, with diameter of inhibition exceeding 41 mm. Moreover, the association between Rosemary EO and the antibiotic (cefoxitin) highlighted the enhancement of the antibacterial effect against the MRSA reference strain. Additionally, Eucalyptus EO showed the highest inhibitory effect against both strains, with MIC values ranging from 0.781 to 1.563 mg/mL, followed by the Rosemary and Lavender EOs. All the tested EOs displayed a bactericidal effect against the tested MRSA strains. Regarding the antibiofilm activity, Rosemary and Lavender EOs had varying impacts on the pre-formed biofilms, with percentage reduction values ranging from 36% to 73% and 37% to 68%, respectively. Finally, the mRNA expression of the MRSA gene A mecA and virulence genes agrA, sarA and sea declined following EO treatment compared with the control. The findings of this study highlighted the efficacy of locally tested EOs in reducing MRSA biofilm formation and the expression of virulence factors and suggested their potential use in food safety and culinary applications. Full article

The spatial organization of the eukaryotic nucleus plays a pivotal role in regulating pre-mRNA splicing; however, the underlying principles governing this organization remain incompletely understood. Recent advances in imaging and sequencing technologies have revealed that splicing regulation is orchestrated across multiple hierarchical levels, from nanoscale protein–RNA interactions to large-scale nuclear architecture. Intrinsically disordered regions (IDRs) in RNA-binding proteins (RBPs) mediate multivalent interactions that drive liquid–liquid phase separation, leading to the formation of dynamic biomolecular condensates, such as nuclear speckles, paraspeckles, and nuclear stress bodies (nSBs). These structures act as functional hubs that modulate RNA processing efficiency and respond to cellular stress. In addition, emerging evidence highlights nucleus-wide RBP meshworks that spatially organize co-transcriptional splicing through dynamic RNA-dependent interactions. The interplay between these condensates and meshworks forms a spatially organized network that fine-tunes the efficiency and fidelity of pre-mRNA splicing. Collectively, this review presents a unified model in which phase separation and higher-order nuclear architecture coordinately regulate transcriptomic output in space and time. Full article

Background/Objectives: Polyethylene glycol (PEG) is widely used to enhance the stability and pharmacokinetics of nanomedicines, including lipid nanoparticle (LNP)-based mRNA vaccines. However, both pre-existing and vaccine-induced anti-PEG antibodies may compromise the efficacy and safety of PEGylated therapeutics. Methods: In this study, we analyzed the specificity and avidity of anti-PEG antibodies in human blood donor samples from Unvaccinated individuals and recipients of PEGylated mRNA-LNP vaccines (Comirnaty and Spikevax), polysorbate-containing vaccines, or vaccines lacking both PEG and polysorbates. Quantitative ELISA was used to characterize anti-PEG and anti-polysorbate IgM and IgG responses in 325 plasma samples, while an equilibrium titration method was applied to assess IgG binding to PEG molecules, micelles, and PEGylated liposomes with defined structural features in 36 plasma samples. Results: Vaccination with PEGylated mRNA-LNPs was associated with increased anti-PEG antibody levels and qualitative changes in antibody binding behavior. Anti-PEG IgG antibodies displayed progressively higher avidity toward larger and structurally more complex PEG-containing antigens, with the strongest binding observed for PEGylated liposomes. Notably, vaccinated individuals, particularly those who received Spikevax, showed increased end-group and backbone-specific avidity, as well as an enhanced ability of antibody paratopes to engage shorter PEG chains. In contrast, polysorbate-containing or PEG-free vaccines did not elicit comparable effects. Conclusions: These findings suggest that vaccination with PEGylated mRNA-LNPs is associated with the emergence of altered antibody populations with increased end-group reactivity and higher avidity toward PEG-directed immune responses, despite PEG being a synthetic, nonprotein polymer. Antibody binding to LNPs, accompanied by the emergence of high-avidity anti-PEG IgG seems to be consistent with an increased risk of adverse events, particularly following repeated vaccinations, including complement activation-related pseudoallergy (CARPA) and anaphylaxis. It may also contribute to altered immune protection against the vaccine target, underscoring the need for avidity-aware risk-benefit assessment of PEGylated therapeutics. Full article

Nipah virus (NiV) is a highly pathogenic zoonotic pathogen. Since its discovery in 1998, recurrent epidemics have occurred in South and Southeast Asia, with a case fatality rate ranging from 40% to 100%. The outbreak in West Bengal, India in early 2026 has once again highlighted its severe threat to public health. To date, no licensed human vaccines or specific therapeutics against NiV are available worldwide. This review systematically summarizes the breakthroughs in antigen design for NiV vaccines, with a focus on conformational stabilization of prefusion F (pre-F) protein, chimeric G/F antigens, and multivalent nanoparticle strategies. In addition, we comparatively analyze the clinical progress of mainstream vaccine platforms, including viral vectors, mRNA and subunit vaccines. Given the sporadic nature and high mortality of NiV infection, the conventional licensing pathway relying on large-scale phase III clinical trials faces substantial practical obstacles. Accordingly, this article discusses adaptive adjustments in regulatory science. We propose several strategies to accelerate the clinical translation and emergency stockpiling of NiV vaccine candidates, including establishing unified correlates of protection thresholds, coordinating multinational regulatory resources, and optimizing the implementation of Animal Rule. Full article

Eastern, Venezuelan, and Western equine encephalitis viruses (EEEV, VEEV, and WEEV) are encephalitic alphaviruses transmitted by mosquitoes throughout the Americas. Infection by these viruses can present in humans as a febrile illness; however, it may progress into potentially life-threatening encephalitis. Currently, no publicly licensed vaccines are available, and at-risk individuals are restricted to superseded vaccines. Here, we will review recent advances in our understanding of how these viruses spread among animal populations and cause disease, and how we can manage their diagnosis and treatment. Additionally, we have summarised the recent developments in vaccines against these viruses in both pre-clinical and clinical stages. Overall, global climate change and ecological disruption drive a need for public access to safe and effective vaccines against EEEV, VEEV, and WEEV, which novel platforms, such as mRNA and viral vectors, may be able to achieve. Full article

The protective effect of arzanol, a natural prenylated phloroglucinol α-pyrone from the Helichrysum microphyllum subsp. tyrrhenicum, was investigated in HaCaT keratinocytes exposed to two inflammatory stimuli: lipopolysaccharide (LPS, 0.5–75 µg/mL), a component of gram-negative bacteria, and polyriboinosinic-polyribocytidylic acid (poly I:C, 0.5–50 µg/mL), a synthetic viral RNA analog. LPS and poly I:C significantly decreased HaCaT cell viability (18–93% reduction in the 5–75 μg/mL LPS range and 25% at 50 μg/mL poly I:C, MTT assay) and increased apoptosis and cell death (NucView 488 and propidium iodide assay) after 3 h and 24 h of exposure. Arzanol (1 h of pre-incubation, 5–25 μM) showed a significant protective effect against LPS and poly I:C-induced damage, preserving cell viability (25% of viability increase at 5 μg/mL LPS concentration, and 30% at 50 μg/mL of poly I:C) and decreasing apoptosis/cell death. Western blot analysis demonstrated the ability of arzanol (5 μM) to reduce the apoptotic protein Bax and the inflammatory cytokine IL-1β levels in HaCaT keratinocytes exposed for 3 h to 5 and 10 μg/mL LPS. Moreover, scratch assay showed the arzanol reparative effect on HaCaT cells. Our results qualified arzanol as a protective drug for dermatological applications in human skin diseases. Full article

The second most prevalent neurodegenerative illness in the world, Parkinson’s disease (PD), currently has no viable treatments. Although it is yet unknown if mitochondrial dysfunction is an initial event or evolves as a result of neurodegeneration, it is thought to be a crucial component of Parkinson’s disease etiology. From the perspective of mitochondrial quality control (MQC), which includes PINK1/Parkin-mediated mitophagy, mitochondrial dynamics, and mitochondrial proteostasis, this article examines mitochondrial dysfunction. Together, these processes preserve mitochondrial homeostasis and prevent the buildup of damaged mitochondria. Dysfunctional mitochondria gradually build up and cause oxidative stress and aberrant cellular signaling when mitochondrial quality control is compromised. According to available data, mitochondrial reactive oxygen species (mtROS) primarily worsen pre-existing mitochondrial damage by encouraging α-synuclein aggregation, cardiolipin remodeling, and dopamine oxidation. In addition, innate immune pathways like cGAS–STING and TLR9 signaling can be triggered by mitochondrial damage-associated molecular patterns (mtDAMPs), especially mitochondrial DNA, which can lead to long-term neuroinflammatory reactions in PD. While new research suggests that m⁶A RNA modification may be involved in the regulation of mitochondrial stress, the PINK1/Parkin pathway is crucial for maintaining mitochondrial homeostasis. Therapeutic approaches that target mitophagy augmentation, neuroinflammatory signaling, and mitochondrial protection have garnered increasing attention. In an attempt to improve mitochondrial function and lessen persistent neuroinflammatory activation, future research will probably need to concentrate on combination treatment techniques. Full article

Background: Spinal Muscular Atrophy (SMA) is a neurodegenerative disease caused by reduced survival motor neuron (SMN) protein levels due to SMN1 gene mutations. The natural history of SMA has dramatically changed since innovative therapies were approved; among them, Risdiplam (an oral molecule) increases the peripheral levels of SMN by modifying the pre-mRNA slicing of the paralogous SMN2 that also codes for the protein. Methods: We performed longitudinal RNA sequencing on peripheral blood samples from 16 adult SMA patients (types II and III) before and after 12 months of Risdiplam treatment to assess transcriptomic changes. Results: During Risdiplam treatment, increased SMN2 transcript levels were observed, which was coherent with the clinical condition of the investigated SMA cohort. Upregulated mitochondria genes or pseudogenes (i.e., MT-ATP8 and MTND1P11) and downregulated autophagy-related pathways were also found. Baseline differences in gene expression between SMA type II and type III involved neurodegenerative (i.e., MS4A3, C4BPA, and NEILS3) and immune-related (B2M) genes. Conclusions: These findings support Risdiplam’s systemic impact in adult SMA subjects and reveal molecular distinctions between SMA phenotypes (types II and III), which may be of some relevance for future clinical and therapeutic strategies. Full article

Metastatic hormone-naïve prostate cancer (mHNPC) is a clinically aggressive form of prostate cancer characterized by early systemic dissemination and poor long-term outcomes; however, the intrinsic epithelial cell states present at diagnosis remain poorly defined. In this study, we performed single-cell transcriptomic profiling of diagnostic prostate biopsy specimens from five patients with treatment-naïve mHNPC using Fixed RNA Profiling. Integrated and case-specific analyses characterized epithelial heterogeneity and lineage-associated transcriptional programs. Across 17,825 high-quality single cells, epithelial heterogeneity was identified in all cases. In addition to luminal androgen receptor (AR)-dependent epithelial cells, reproducible basal-associated epithelial populations with reduced AR signaling and stem-like transcriptional features were observed across tumors. Epithelial–mesenchymal transition (EMT)-related transcriptional programs were detected across multiple epithelial states with inter-case variability without forming a distinct EMT cluster, whereas no transcriptionally discrete neuroendocrine epithelial cluster was identified at baseline. These findings demonstrate that treatment-naïve mHNPC harbors pre-existing basal-associated epithelial states that contribute to intrinsic tumor heterogeneity at diagnosis. The presence of AR-low and stem-like epithelial populations prior to systemic therapy suggests a potential role for lineage plasticity in the aggressive biological behavior of metastatic prostate cancer. Full article

Splicing factor 3b1 (SF3B1), a component of U2 small nuclear ribonucleoprotein (U2 snRNP), has been known for its essential roles in pre-mRNA splicing and alternative splicing. Here we show that knocking down (KD) of SF3B1 broadly induced a significant reduction in mRNA expression in the genome. One of the genes whose expression is reduced by SF3B1 KD is methyl-transferase-like 3 (METTL3), a writer of N6-methyladenosine (m⁶A). We demonstrate that expression of both METTL3 mRNA and protein is affected by SF3B1 KD, which further decreases the m⁶A RNA expression level. m⁶A-seq indicates that SF3B1 KD affects m⁶A distribution within multiple genes in the genome. In addition, a high proportion of hypo-methylation events by SF3B1 KD (~70%) are overlapped in METTL3 KD cells, and a conserved m⁶A motif is observed in the hypo-methylated regions as in SF3B1 KD cells, suggesting the m⁶A decrease by SF3B1 is a direct effect of the reduced METTL3 expression. Furthermore, RT-qPCR using unlabeled RNA and 5-Bromouridine (BrU)-labeled nascent RNA and actinomycin D treatment demonstrates that transcription of METTL3 is significantly reduced but the mRNA decay rate is not altered, suggesting that METTL3 expression is altered at the transcription level. We further show that SF3B1 interacts with RNA polymerase (Pol) II in the RNA independent manner, further indicating the involvement of SF3B1 in transcription. Lastly, we demonstrate that the transcription inactive H3K27me3 on the METTL3 promoter was significantly increased whereas transcription active H3K4me3 was not changed by SF3B1 KD. Taken together, we conclude that reduced SF3B1 expression suppresses the transcription of METTL3 and inhibits m⁶A RNA expression. Full article

Background: Salinity, which hampers wheat growth and development, is one of the major abiotic stresses. Plant-derived smoke (PDS) solution alleviates salt stress and promotes wheat growth and development; however, the underlying molecular mechanisms have not been completely clarified. Methods: In this study, nuclear proteomics was employed to reveal the promotive effect of PDS solution on salt-stressed wheat. Nuclear fractions were isolated from wheat roots, and their purity was confirmed via enrichment of histone H3 and reduction of cytosolic ascorbate peroxidase. Using this nuclear purification technique, label-free nano LC–MS/MS-based nuclear proteomics was performed to identify differentially abundant nuclear proteins in salt-stressed wheat with or without PDS solution treatment. Results: Salt stress decreased histone H2A and DNA polymerase levels, whereas PDS solution treatment of salt-stressed wheat increased levels of histone variants (H2A, H2B, H3, and H4), DNA polymerase, and DNA topoisomerase II. In addition, the PDS solution increased the levels of pre-mRNA cleavage factor Im 25 kDa subunit and RNA helicase in salt-stressed wheat. Immunoblot analysis further validated the increase in histone deacetylase levels triggered by the PDS solution treatment in the salt-stressed wheat. Conclusions: These results suggest that PDS solution alters nuclear proteins in a way that contributes to chromatin remodeling and transcription during salt stress. Full article

Background/Objectives: MicroRNAs are key post-transcriptional regulators involved in various diseases. Despite its status as the gold standard, real-time RT-PCR faces challenges arising from high sequence homology among closely related microRNAs and the substantial biomaterial required to enrich small RNA fractions. This study aimed to develop an optimized protocol for simultaneous analysis of microRNA and mRNA expression from a single total RNA sample using mouse (Mus musculus) brain tissue, avoiding dependence on pre-designed commercial assay panels. Methods: We optimized a real-time RT-PCR workflow enabling simultaneous analysis of mature microRNAs and mRNAs from a single total RNA sample. Modifications include a redesigned universal reverse primer, LNA-modified TaqMan probes, and omission of the 65 °C denaturation step during reverse transcription. The method was validated for five microRNAs in mouse brain tissue. Results: The assay showed high specificity, discriminating closely related miR-125a-5p and miR-125b-5p with a ΔCt difference of 6.7 ± 1.2 cycles. Co-analysis with Oligo(dT)₁₈ and Random hexamer primers did not interfere with microRNA detection. Conclusions: The developed approach enables reliable detection of closely related microRNAs and parallel analysis of different RNA types, which is particularly important for studying regulatory networks when working with limited amounts of biomaterial. This protocol provides a complementary, accessible option for targeted studies in resource-limited settings or for non-cataloged miRNA targets. Full article

Background/Objectives: Barley malting is an agro-industrial process that produces malt, an essential ingredient for the brewing and distilling industries. Previously, tran-scriptome profiling has revealed mRNA changes during malting but less is known about their regulation. Methods: The spring 2-row barley variety ‘Conrad’ was sampled at five stages of malt-ing. Using small RNA (sRNA)-sequencing and degradome-sequencing data from these malting stages, de novo discovery of mature microRNA (miRNA), as well as cognate mRNAs targeted for slicing, was identified. ShortStack v4.1.0 was used to map sRNA reads to the Hordeum vulgare Morex V3 genome. Results: In total, 33 expressed MIRs were identified, six of which may be novel. Using the degradome-sequencing data from the same malting stages, CleaveLand4 v4.5 pre-dicted 64 sliced mRNA targets, predominantly transcription factors associated with root development. Conclusions: This study provides an overview of post-transcriptional modulations of miRNAs-cognate mRNA targets, as well as plausible interactions between miRNAs during barley malting. Full article

Background/Objectives: MicroRNAs (miRNAs) are key regulators of skeletal muscle adaptation; however, the extent to which exercise modulates the miRNA biogenesis pathway remains poorly understood. To investigate the impact of acute and chronic high-intensity exercise on components of miRNA biogenesis, and whether such changes are reflected in miRNA expression across stages of their biogenesis, we performed secondary analyses of muscle biopsy samples from two previously published studies. Methods: Muscle biopsies were analyzed from the following protocols: nine men and eight women pre- and 3 h post- a bout of high-intensity interval cycling exercise (HIIE), and eleven men and eight women pre- and post- a 6-week period of high-intensity interval training (HIIT) or non-exercise control. mRNA expression of components of miRNA biogenesis including Drosha, Exportin-5, Dicer, and Ago2 were assessed following HIIE using RT-qPCR and their protein abundance was measured following HIIT using Western blotting. Primary (pri-miR-133a1, -133a2, -133b) and mature (miR-133a-3p, -133a-5p, -133b) miRNA expression were quantified following HIIT. Results: An acute bout of HIIE significantly decreased Drosha mRNA (p < 0.05) and resulted in a reduction in Dicer mRNA that approached significance (p < 0.10). Following 6 weeks of HIIT, no significant changes were detected in the protein abundance of Drosha, Exportin-5, Dicer, or Ago2. HIIT did not alter miR-133 expression at either the primary or mature transcript level across all isoforms. Conclusions: This study highlights the complexity of miRNA regulation in skeletal muscle and underscores the need for further research examining the temporal and mechanistic control of miRNA biogenesis in response to exercise. Full article

Background: Acetamiprid (ACMP) exposure mediates a variety of pathological complications, including testicular toxicity. Berberine (BBR) is a plant-derived alkaloid with potential pharmacological properties. This study sought to evaluate the ameliorative effects of BBR against ACMP-induced testicular toxicity. Methods: Male Wistar rats were divided into four groups: control, BBR-treated, ACMP-exposed, and BBR+ACMP co-treated, and were administered with BBR (150 mg/kg b.wt) and ACMP (21.7 mg/kg b.wt) for 21 days. Biochemical and FTIR analyses, RT-PCR, computational analyses, and histopathological examination were conducted to assess alterations in lipid and protein profiles, as well as apoptotic and structural changes. Results: ACMP exposure was associated with oxidative injury, functional alterations (stretching of -OH, -CH₂, -NH, C=O, C-N, -COO-, -PO₂⁻), and compositional changes in proteins and lipids. Pre-treatment of BBR (2 h prior) was associated with attenuation of the functional and compositional alterations in proteins and lipids in co-treated rats. RT-PCR and computational analysis showed increased Bax and caspase-3 and decreased Bcl-2 mRNA expression, suggesting a potential modulation of ACMP-induced apoptosis by BBR. Histological examination showed that pre-treatment with BBR prevented ACMP-induced structural alterations, including cellular disorganization and alteration in seminiferous tubules. Conclusions: The study suggested that the BBR may exert ameliorative effects against ACMP-induced testicular toxicity by modulating lipid and protein changes and the anti-apoptotic pathway. Thus, BBR could be used as a potential ameliorative agent against oxidative stress. However, more mechanistic studies are needed for broader biological relevance and validity. Full article