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This study investigated the wound healing properties of the aqueous extract of Morinda tinctoria Roxb leaves in rats. The study also provides information on the purification, qualitative, and quantitative analysis of phytochemical components present in M. tinctoria Roxb. Wound contraction and period of epithelialization was determined and topical application of M. tinctoria Roxb aqueous leaf extract showed better healing than orally treated and control groups. The results suggest that the efficacy of M. tinctoria Roxb aqueous leaf extract as a wound healing agent and can also be used as a therapeutic agent for internal as well as external wounds.

Background Sequence and structural elements in the 3'-untranslated region (UTR) of Japanese encephalitis virus (JEV) are known to regulate translation and replication. We previously reported an abundant accumulation of small subgenomic flaviviral RNA (sfRNA) which is collinear with the highly conserved regions of the 3'-UTR in JEV-infected cells. However, function of the sfRNA in JEV life cycle remains unknown. Results Northern blot and real-time RT-PCR analyses indicated that the sfRNA becomes apparent at the time point at which minus-strand RNA (antigenome) reaches a plateau suggesting a role for sfRNA in the regulation of antigenome synthesis. Transfection of minus-sense sfRNA into JEV-infected cells, in order to counter the effects of plus-sense sfRNA, resulted in higher levels of antigenome suggesting that the presence of the sfRNA inhibits antigenome synthesis. Trans -acting effect of sfRNA on JEV translation was studied using a reporter mRNA containing the luciferase gene fused to partial coding regions of JEV and flanked by the respective JEV UTRs. In vivo and in vitro translation revealed that sfRNA inhibited JEV translation. Conclusions Our results indicate that sfRNA modulates viral translation and replication in trans .

Chronic wounds present a major clinical challenge, often aggravated by infection and rising antimicrobial resistance. This study investigated the wound-healing efficacy of Lobelia alsinoides Lam., an ethnomedicinal herb, formulated as a topical ointment containing its ethanolic extract (LT). Phytochemical profiling identified high levels of phenolics, terpenoids, and tannins, while in vitro assays demonstrated strong antioxidant, broad-spectrum antimicrobial, and cytocompatible properties. Wound-healing potential was evaluated using excision and incision wound models in rats treated with 5% or 10% LT ointments, with Silverex™ as the reference standard. The 10% LT formulation significantly outperformed Silverex™, accelerating wound contraction (99.33 ± 0.55% by Day 16), shortening epithelialization time (16.1 ± 0.8 days), and enhancing tensile strength (837.36 ± 16.37 g; p < 0.001). Biochemical and histological analyses confirmed improved collagen deposition, extracellular matrix remodeling, and angiogenesis, without hepatic or renal toxicity. Overall, LT exhibited statistically superior wound-healing efficacy compared with Silverex™, supporting its potential as a safe, affordable, and sustainable phytotherapeutic alternative. These findings provide strong scientific validation for L. alsinoides as an evidence-based herbal candidate for integration into modern wound care, with future studies warranted to establish mechanistic and clinical efficacy in chronic and infected wounds.

Classical swine fever virus (CSFV) causes a broad range of disease in pigs, from acute symptoms including high fever and hemorrhages, to chronic disease or unapparent infection, depending on the virus strain. CSFV belongs to the genus Pestivirus of the family Flaviviridae. It carries a single-stranded positive-sense RNA genome. An internal ribosomal entry site (IRES) in the 5' untranslated region (UTR) drives the translation of a single open reading frame encoding a 3898 amino acid long polypeptide chain. The open reading frame is followed by a 3' UTR comprising four highly structured stem-loops. In the present study, a synthetic RNA composed of the 5' and 3' UTRs of the CSFV genome devoid of any viral coding sequence and separated by a luciferase gene cassette (designated 5'UTR-Luc-3'UTR) triggered apoptotic cell death as early as 4 h post-transfection. The apoptosis was measured by DNA laddering analysis, TUNEL assay, annexin-V binding determined by flow cytometry, and by analysis of caspase activation. Contrasting with this, only trace DNA laddering was observed in cells transfected with the individual 5' or 3' UTR RNA; even when the 5' UTR and 3' UTR were co-transfected as separate RNA molecules, DNA laddering did not reach the level induced by the chimeric 5'UTR-Luc-3'UTR RNA. Interestingly, RNA composed of the 5'UTR and of stem-loop I of the 3'UTR triggered much stronger apoptosis than the 5' or 3'UTR alone. These results indicate that the 5' and 3' UTRs act together in cis induce apoptosis. We furthered obtained evidence that the UTR-mediated apoptosis required double-stranded RNA and involved translation shutoff possibly through activation of PKR.

Background Classical swine fever virus (CSFV) is the member of the genus Pestivirus under the family Flaviviridae . The 5' untranslated region (UTR) of CSFV contains the IRES, which is a highly structured element that recruits the translation machinery. The 3' UTR is usually the recognition site of the viral replicase to initiate minus-strand RNA synthesis. Adenosine-uridine rich elements (ARE) are instability determinants present in the 3' UTR of short-lived mRNAs. However, the presence of AREs in the 3' UTR of CSFV conserved in all known strains has never been reported. This study inspects a possible role of the ARE in the 3' UTR of CSFV. Results Using RNA pull-down and LC/MS/MS assays, this study identified at least 32 possible host factors derived from the cytoplasmic extracts of PK-15 cells that bind to the CSFV 3' UTR, one of which is HuR. HuR is known to bind the AREs and protect the mRNA from degradation. Using recombinant GST-HuR, this study demonstrates that HuR binds to the ARE present in the 3' UTR of CSFV in vitro and that the binding ability is conserved in strains irrespective of virulence. Conclusions This study identified one of the CSFV 3' UTR binding proteins HuR is specifically binding to in the ARE region.

Drought stress poses a significant threat to fertile soils worldwide, triggering profound physiological, biochemical, and molecular changes in plants that adversely impact agricultural productivity. This study explores the potential of nanotechnology, specifically Calcium Oxide Nanoparticles (CaO NPs) and Graphene Oxide (GO), to ameliorate the negative effects of drought stress on two distinct alfalfa ecotypes. Seeds from Erzurum and Konya regions were regenerated in the Murashige and Skoog (MS) medium, and ensuing callus formation was induced through 1 mg L−1 2,4-D and 1 mg L−1 kinetin MS medium. The callus samples underwent a one-month treatment with varying concentrations of mannitol (50 and 100 mM), CaO NPs, and GO (0.5 and 1.5 ppm). Results revealed a decrease in dry/wet weight with increasing mannitol concentration, contrasting with an increase in weight under CaO NPs and GO treatment. Proline, DNSA, MDA, and H2O2 exhibited proportional increases under drought stress, while CaO NPs and GO treatments mitigated these effects. Physiological and biochemical analyses identified optimal conditions for Erzurum as 50 mM mannitol/2 CaO NPs/0.5 ppm GO, and for Konya as 50 mM mannitol/0.5 ppm GO. Gene expression analysis indicated up-regulation of mtr-miR159 and mtr-miR393 with heightened drought stress, with down-regulation observed in CaO NPs and GO treatments. Scanning Electron Microscopy (SEM) and Confocal Laser Scanning Microscopy (CLSM) confirmed Ca2+ accumulation in alfalfa tissues. In conclusion, CaO NPs and GO treatments exhibited a significant reduction in the adverse effects of drought stress on alfalfa callus under tissue culture conditions. This research sheds light on the potential of nanotechnological interventions to alleviate the impact of environmental stressors on crop plants, opening avenues for sustainable agriculture in the face of changing climatic conditions. Further investigations are warranted to elucidate the underlying mechanisms and scalability of these findings for field applications.Key messageCaO NPs and GO enhanced the tolerance of M. sativa callus under drought stress improving biochemical activity, miRNA gene expression, confocal laser scanning, and electronic scanning analysis.

[This corrects the article DOI: 10.1371/journal.pone.0088863.].

The Lobelia alsinoides Lam plant has traditionally been consumed as a wild food supplement by rural communities in Thailand’s north-eastern Kalasin province. Despite its traditional use, there has been a lack of scientific research to validate Lobelia alsinoides Lam’s biological properties. This is the first study to identify and characterize the ethanolic extract of Lobelia alsinoides Lam and assess it’s in vitro cytotoxic potential. Various spectroscopic techniques, including infrared (IR) spectroscopy, NMR spectroscopy ( 1 H and 13 C), and mass spectroscopy (LC-QTOF-MS), were used to characterize the Lobelia alsinoides Lam ethanolic extract. The extract was then tested for cytotoxicity against various cancerous cell lines (A549, KB, HeLa, MDA-MB-231, and MCF-7). Spectral investigation on the ethanolic extracts of the Lobelia alsinoides Lam’s plant afforded four components namely 5-{2-[(3,4-Dihydroxy-5-methyl-phenyl)-methyl-amino]-propoxy}-4,6-dimethyl-tetrahydro-pyran-2,3-diol ( 1 ), (3,4-Dimethyl-phenyl)-methyl-carbamic acid 3,4,5-trihydroxy-6-methyl-tetrahydropyran-2-yl ester ( 2 ); 5-{[3-(2,6-Dimethyl-tetrahydro-pyran-3-yl)-1-methyl-propyl]-methyl-amino}-benzene-1,2,3,4-tetraol ( 3 ); 3-Hydroxy-5-[3,4,5-trihydroxy-6-(3,4,5-trihydroxy-6-methyl-tetrahydro-pyran-2-yloxymethyl)-tetrahydro-pyran-2-yloxy]-pent-3-enoic acid methyl-(2,3,4,5-tetrahydroxy-phenyl)-amide ( 4 ). The ethanol extract of Lobelia alsinoides Lam was found to have a significant cytotoxic effect in a dose-dependent manner against five human cancer cells. The LDH release assay and the NO inhibitory assay confirmed Lobelia alsinoides Lam’s anti-proliferative and anti-inflammatory effects in vitro compared with the standard drug cisplatin. According to our findings, the wild food source Lobelia alsinoides Lam is non-toxic and may have cytotoxic activity against human cancers such as lung, breast, cervical, and oral. Graphical Abstract Highlights First study to report the characterization of ethanolic extract of Lobelia alsinoides Lam using FTIR, NMR ( 1 H and 13 C), and mass spectroscopy (LCMS-QTOF-MS). L. alsinoides Lam ethanol extract was found to have a potent cytotoxic effect in dose-dependent response against the five human cancer cells (A549, KB, HeLa, MDA-MB-231, and MCF-7) LDH release assay, and the NO inhibitory assay, confirmed the in vitro antiproliferative and anti-inflammatory effects of L. alsinoides Lam

Classical swine fever virus (CSFV) causes a broad range of disease in pigs, from acute symptoms including high fever and hemorrhages, to chronic disease or unapparent infection, depending on the virus strain. CSFV belongs to the genus Pestivirus of the family Flaviviridae. It carries a single-stranded positive-sense RNA genome. An internal ribosomal entry site (IRES) in the 5' untranslated region (UTR) drives the translation of a single open reading frame encoding a 3898 amino acid long polypeptide chain. The open reading frame is followed by a 3' UTR comprising four highly structured stem-loops. In the present study, a synthetic RNA composed of the 5' and 3' UTRs of the CSFV genome devoid of any viral coding sequence and separated by a luciferase gene cassette (designated 5'UTR-Luc-3'UTR) triggered apoptotic cell death as early as 4 h post-transfection. The apoptosis was measured by DNA laddering analysis, TUNEL assay, annexin-V binding determined by flow cytometry, and by analysis of caspase activation. Contrasting with this, only trace DNA laddering was observed in cells transfected with the individual 5' or 3' UTR RNA; even when the 5' UTR and 3' UTR were co-transfected as separate RNA molecules, DNA laddering did not reach the level induced by the chimeric 5'UTR-Luc-3'UTR RNA. Interestingly, RNA composed of the 5'UTR and of stem-loop I of the 3'UTR triggered much stronger apoptosis than the 5' or 3'UTR alone. These results indicate that the 5' and 3' UTRs act together in cis induce apoptosis. We furthered obtained evidence that the UTR-mediated apoptosis required double-stranded RNA and involved translation shutoff possibly through activation of PKR.