Search Results (9,701)

Effective in vitro propagation of medicinal and aromatic plants such as Thymus vulgaris is often limited by the choice of optimal cytokinin sources, which are critical for successful shoot proliferation and overall regeneration. Since their discovery, topolins have been recognized as alternatives to traditional cytokinins (CKs) such as benzyladenine (BA) and kinetin (Kin) in plant tissue culture (PTC). This study investigated the influence of three aromatic CKs (meta-topolin (mT), BA, and Kin), on shoot proliferation of thyme, with the goal of improving current PTC protocols. Of all the tested treatments, the highest shoot proliferation (7.25 ± 0.72 shoots per nodal explant) was observed in the treatment with 1 µM mT, superior to BA and Kin. Increasing mT and BA concentrations from 1 to 15 µM had an inverse effect on shoot production. The addition of indole-3-butyric acid (IBA) at concentrations ranging from 1.0 to 5.0 µM in combination with 0.5 and 1 µM mT did not increase the mean shoot numbers per explant. Regenerated shoots exhibited a strong propensity for root development even in the absence of plant growth regulators with 100% survival ex vitro. This study demonstrated that mT is an effective, sustainable and cost-effective alternative to traditional CKs for in vitro propagation of T. vulgaris, achieving an improved shoot proliferation with 1 μM mT application. Full article

Dimethyl fumarate (DMF) has established a significant position among therapies for multiple sclerosis and psoriasis and is now being investigated for repurposing to many other non-malignant diseases. Despite decades of preclinical research, some issues about its pharmacology remain unresolved, with ongoing debate over which of the methyl esters of fumarate, whether DMF or monomethyl fumarate (MMF), is the active ingredient. It is generally accepted that DMF undergoes enzymatic hydrolysis to MMF and methanol. However, there is disagreement regarding its exact site, its extent, and the responsible enzyme(s). The enzymatic mechanisms, particularly the roles of carboxylesterases-1 and 2, vary across tissues and species, complicating the translation of in vitro and in vivo preclinical findings into clinical practice. In addition, the impact of DMF and MMF is often not clearly distinguishable and sometimes overlaps, making the true molecular mediators of therapeutic and side effects unclear. Thus, the interpretation of some results obtained in studies is inconsistent because of interchanging of in vitro and in vivo observed features of fumarate esters: while DMF demonstrates rapid and strong effects in cell culture studies, including nuclear factor erythroid 2-related factor 2 (NRF2) function activation and glutathione depletion, these observations may not exactly reflect systemic pharmacology and physiology dominated by MMF. Moreover, methanol, the co-product of DMF metabolism, may contribute to the observed DMF effects through increased production of reactive oxygen species, which could result in activation of NRF2-dependent mechanisms. This review highlights specific unresolved issues in DMF metabolism, which are sometimes overlooked. Full article

To identify the primary pathogen responsible for brown rot in mature Korla fragrant pears and to screen for effective biocontrol bacterial strains, a research team collected 30 samples of brown rot-infected fruits and 84 samples from healthy trees (including branches, leaves, and fruits) from three Korla pear cultivation areas in Korla, Aksu, and Kashgar between September and October 2024. Investigation into the severity of the disease revealed the local field incidence rate of brown rot ranged from 17% to 31%. From the 30 disease samples, 23 morphologically identical fungal strains were isolated, with 13 strains isolated from Korla City (56.5%), 6 from Aksu City (26.1%), and 4 from Tumushuk (17.4%), all detected in infected fruits. Pathogenicity tests were conducted using both inoculation by wounding with mycelium and spray inoculation experiments to study the pathogens’ effect on brown rot in Korla fragrant pears. From the 84 samples of healthy trees, 55 bacterial strains were isolated, and the antagonistic bacteria’s inhibitory effect on the isolated pathogens was determined using the dual-culture method. The pathogen was identified by morphological characteristics and phylogenetic analysis based on multi-locus sequencing (ITS-TUB2-Lcc2). Antagonistic bacterial strains were identified through morphological observation and 16S rDNA sequencing. The results demonstrated that the pathogen isolated from the diseased Korla fragrant pear tissues was identified as Monilinia yunnanensis. Among the biocontrol antagonistic bacteria isolated from the branches, bark, and leaves of healthy Korla fragrant pear trees, Bacillus siamensis exhibited significant inhibitory effects against the pathogen, with an in vitro inhibition rate of (88.18 ± 3.43)%. Full article

(1) Background: Low-molecular-weight hyaluronic acid displays moisturizing and anti-aging properties and reduces UV-induced inflammation when applied topically. A 3 kDa sodium hyaluronate oligosaccharide (Extra-Low HA) was designed, and studies were performed to evaluate its safety for cosmetic applications. (2) Methods: The ability of the Extra-Low HA (ExLMW-HA) to penetrate skin was evaluated. Then, pro-inflammatory cytokines were quantified in the culture medium of skin explants following ExLMW-HA application with or without inflammation inducer (PMA). Finally, four predictive in vitro tests (Keratinosens^™, kDPRA, Ames’ test, micronucleus test) were conducted to assess the safety of ExLMW-HA. (3) Results: The molecule permeates skin down to the living epidermis and possibly interacts with the dermal compartment. The oligosaccharide did not induce TNF-α, IL-1β, IL-1α, CXCL2, CCL3, or IL-15, neither in basal nor in stressed conditions. ExLMW-HA is not predicted to be a skin sensitizer or a mutagenic or genotoxic substance. (4) Conclusions: This 3 kDA HA is considered safe for use in topical application at the tested dosage. Full article

Mitochondria are crucial for energy metabolism and the regulation of apoptosis and the inflammatory response in acute myeloid leukemia (AML). This study examined key mitochondrial characteristics in apoptosis-resistant AML cells during in vitro aseptic pro-inflammatory activation utilizing spectrofluorimetry, quantitative reverse transcription PCR, Western blotting, differential gene expression analysis, flow cytometry, transmission electron microscopy, and cellular respiration analysis. Under conditions of aseptic inflammation simulated in three-dimensional high-density cultures, apoptosis-resistant AML cells exhibited a significant reduction in the transcriptional activity of genes linked to oxidative phosphorylation and the tricarboxylic acid cycle; demonstrated diminished mitochondrial respiration activity; and decreased levels of the mitophagy regulatory proteins PINK1 and Parkin. Furthermore, pathogenic alterations in mitochondrial morphology were observed. These cells demonstrated enhanced intracellular generation of reactive oxygen species, lactate accumulation in the culture media, elevated levels of DRP1 protein, and an increased fraction of small and medium-sized mitochondria. The acquired data demonstrate that aseptic pro-inflammatory activation results in metabolic remodelling of acute myeloid leukemia cells, integrating characteristics of mitochondrial dysfunction. This condition may facilitate the persistence of leukemic cells during inflammatory stress and potentially contribute to the development of an apoptosis-resistant phenotype. The established in vitro model is crucial for examining both the characteristics of energy metabolism and the anti-apoptotic mechanisms in leukemic cells. Full article

Platelet shortage poses a significant barrier to research and transfusion therapies because native megakaryocytes (MKs) are scarce in blood. To overcome this limitation, pluripotent stem cell–derived MKs (PSC-MKs) offer a standardized, donor-independent platform for research and therapeutic development, including disease modeling and ex vivo platelet production. Here, we report a chemically defined, feeder-free protocol to generate MKs from human pluripotent stem cells (hPSCs). The protocol combines the small molecule MPL agonist Butyzamide, macrophage colony-stimulating factor (M-CSF), and three-dimensional (3D) suspension culture, achieving high efficiency and reproducibility. Butyzamide replaced recombinant thrombopoietin (TPO), yielding comparable CD41⁺/CD42b⁺ populations and enhanced polyploidization. M-CSF accelerated nuclear lobulation and induced 4N MKs, while 3D culture increased yield, cell size, and substrate detachment. Multiple independent assays confirmed mature MK hallmarks, multi-nuclei, demarcation membranes, granules, and elevated mitochondrial respiration. Single-cell RNA sequencing outlined a continuous trajectory from early progenitors to functionally specialized MK subsets. This platform enables reliable MK supply for mechanistic studies and in vitro platelet production, advancing both basic research and therapeutic development. Full article

Effective communication between multipotent mesenchymal stromal cells (MSCs) and endothelial cells (ECs) plays a critical role in the regulation of angiogenesis, especially under conditions of hypoxia. In addition to paracrine stimulation, direct intercellular contacts play an important role in the angiogenic interaction between MSCs and ECs, making them an important target for modulating vascular network restoration under ischemic conditions. The aim of this study was to determine the contribution of gap junctions (GJs) to the angiogenic response of MSCs and the EA.hy926 cell line (an Endothelial Cell Model) under acute hypoxic stress. In a cell co-culture model at 0.1% O₂ using a specific GJ inhibitor (carbenoxolone), molecular, cellular, and functional tests were performed: assessment of viability, proliferation, migration, secretion of angiogenic mediators, and expression of crucial genes. GJ blockade was accompanied by decreases in the proliferation and migration activity and angiogenic potential of the conditioned medium in in vitro and in ovo tests. These data highlight the importance of the GJ in coordinating the angiogenic response in conditions of acute hypoxia and can be used to develop protocols for regenerative medicine. Full article

Porlieria chilensis Johnst. (guayacán), an endemic Chilean species native to the sclerophyllous forests, is experiencing a significant population decline. Typically growing as a shrub or small tree on sunny slopes and rocky soils, its populations have been severely impacted by overexploitation and habitat degradation, leading to its classification as Vulnerable (VU). Mature, well-developed individuals have become increasingly rare, now mostly replaced by shrubby regrowth. This study presents the first reported in vitro propagation method for P. chilensis, aimed at establishing a protocol for its mass propagation. A 92.0% survival rate was achieved for nodal segments cultured on Murashige and Skoog (MS) medium after surface disinfection with 1.5% sodium hypochlorite for 15 min. Shoot elongation was successfully stimulated by supplementing the medium with 8.88 μM of 6-benzylaminopurine (BAP), resulting in an average shoot length of 6.9 cm. For ex vitro rooting, plants were transferred to 200 mL containers filled with a 2:1 (v/v) peat/perlite substrate. High rooting rates were obtained (87.5% and 93.1%) when treated with 1968 μM and 1476 μM of indole-3-butyric acid (IBA), respectively. Moreover, 89.8% of the plants survived acclimatization under controlled conditions. These results highlight the strong potential of in vitro propagation as an effective strategy for the conservation and restoration of P. chilensis populations. Full article

Rice sheath blight caused by Rhizoctonia solani is a devastating global rice disease. This study aimed to isolate biocontrol bacteria from the medicinal plant Bletilla striata for managing the disease. Strain A1 demonstrated the strongest antagonistic activity, with a 91.92% inhibition rate against R. solani in vitro. It also exhibited a broad antifungal spectrum against ten plant pathogenic fungi. Morphological and molecular (16S rRNA and recA genes) analysis identified strain A1 as Pseudomonas koreensis. In detached leaf assays, lesion length was significantly reduced. Pot and field trials showed control efficacies of 65.54% and 72.53%, respectively, comparable to the chemical agent Jinggangmycin. Strain A1 secreted extracellular enzymes (protease, β-1,3-glucanase), siderophores, and auxin (IAA), and possessed phosphate-solubilizing and nitrogen-fixing capabilities. The strain significantly enhanced the activities of key defense enzymes (POD, PAL, PPO, CAT, SOD) in rice. Furthermore, both its sterile culture filtrate and the corresponding crude ethyl acetate extract exhibited strong, direct suppression of R. solani growth. LC-MS analysis identified potential antifungal compounds, including Pseudomonic Acid, Artemisinin, and Tetradecane, in the extract. In conclusion, P. koreensis A1 is a promising biocontrol and plant growth-promoting candidate for sustainable management of rice sheath blight. Full article

Globba sirirugsae Saensouk & P.Saensouk, known in Thai as Hong Hoen Sirirugsa, is a rare Zingiberaceae species with considerable potential for ornamental horticulture and phytopharmaceutical development. Despite its promising attributes, comprehensive studies on its micropropagation, bioactivities, and phytochemical composition remain limited. This study investigated the efficiency of in vitro propagation using rhizome-derived plantlets cultured on Murashige and Skoog (MS) medium supplemented with various concentrations of BA, kinetin, and NAA. The highest shoot proliferation (5.67 shoots) was achieved with 4 mg/L BA and 0.5 mg/L NAA, while acclimatization in a soil–sand substrate (1:1) resulted in a 90% survival rate. Comparative analyses of wild and tissue-cultured plants revealed abundant phenolic and flavonoid contents, particularly in wild specimens, as determined by TPC and TFC assays. HPLC profiling confirmed the presence of bioactive compounds under both growth conditions. Ethanolic extracts exhibited strong antioxidant activities via 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays. GC-MS analysis identified 23 volatile compounds in wild plants and 51 in tissue-cultured plants, with α-pinene, β-pinene, caryophyllene, and α-bergamotene as dominant constituents. FTIR spectroscopy revealed distinct functional groups and fingerprint regions, serving as a rapid screening tool for phytochemical accumulation and biological activity. These findings provide a strategic foundation for the conservation and sustainable utilization of Globba sirirugsae as a multifunctional bioresource, with future applications in pharmaceutical innovation, product development, and ornamental landscaping. Full article

The absence of a high-efficiency and stable genetic transformation system has been a critical bottleneck, impeding both functional gene characterization and precision breeding efforts in Camellia sinensis (tea), and browning is the first problem encountered in tissue culture of tea. In this paper, to identify optimal spectral conditions for minimizing browning in tissue culture, we subjected three tea plant cultivars to distinct light quality treatments and conducted comprehensive metabolomic profiling of their phytochemical contents. This study demonstrates that wavelength-specific light treatments can induce reversible modifications in the physicochemical characteristics of tea leaves, effectively reducing the accumulation of flavonoid compounds, including polyphenols, in plant tissues. Notably, tissues subjected to optimized wavelength conditions exhibit superior performance as explant sources for in vitro culture systems, demonstrating significantly lower browning rates. Comparative analysis of 460 nm, 660 nm, and 730 nm irradiation treatments revealed consistent suppression of polyphenol biosynthesis across all examined cultivars. However, the wavelength eliciting maximal reduction showed significant cultivar-dependent variation, indicating genotype-specific photoresponsive regulation of secondary metabolism. Full article

Background/Objectives: Skeletal muscle plays a pivotal role in whole-body glucose metabolism and is a major target in the pathogenesis and treatment of insulin resistance and type 2 diabetes. The C2C12 myotube cell line is one of the most used in vitro models to investigate mechanisms of insulin resistance. This systematic review (1) summarizes the most common experimental conditions including palmitate concentrations and treatment durations used to induce insulin resistance in C2C12 myotubes; (2) characterizes outcomes related to insulin resistance; and (3) discusses strengths and limitations associated with this model. Methods: A systematic search of PubMed and Scopus was conducted using terms “C2C12 AND palmitate AND insulin resistance” and related variations. A total of 191 articles met inclusion criteria. Results: The most frequently used palmitate concentrations were 0.25 mM, 0.5 mM, and 0.75 mM for at least 16 h, which consistently led to decreased insulin-stimulated pAkt expression, GLUT4 abundance, and insulin-stimulated glucose uptake. Conclusions: The high volume and consistency of primary findings is a key strength of this article which demonstrated reduced insulin signaling across various culture conditions, treatment durations, and insulin co-stimulation protocols. Full article

With emerging viruses and drug resistance on the rise, the discovery and development of innovative antiviral substances and agents are necessary for the effective treatment and control of viral outbreaks. Surrogate viruses are safer alternatives used in research to mimic dangerous or hard-to-culture viruses. They enable efficient, ethical, and cost-effective screening of antiviral compounds. In this study, we used a recombinant viral hemorrhagic septicemia virus (rVHSV) expressing enhanced green fluorescent protein as a surrogate for RNA viruses for the high-throughput screening of antiviral agents. An optimized mixture of viruses and EPC host cells was distributed in 96-well plates containing chemical compounds or plant extracts for screening. Using this system, 44,642 chemical compounds and 8104 plant and marine organism extracts were tested; 140 candidates were selected from primary screening, and 8 compounds and 5 plant extracts were further selected based on the selectivity index (SI), representing the ratio of the cytotoxic concentration (CC₅₀) to the inhibition concentration (IC₅₀). Among these, compound 3, which had the highest SI value of 1046, was further tested, considering in vitro activity against VHSV and another fish rhabdovirus, snakehead rhabdovirus (SHRV). Full article

Background/Objectives: Endothelial cells play a key role in peripheral nerve regeneration, forming aligned vasculature which bridges the gap in the injured nerve tissue and guides the regrowing tissue. This work aimed to mimic key features of this aligned vasculature by differentiating endothelial cells from human induced pluripotent stem cells (hiPSCs) and incorporating them into engineered neural tissue (EngNT). Methods: hiPSCs were differentiated into endothelial cells with the temporal addition of growth factors and biomolecules. These hiPSC-derived endothelial cells (hiPSC-ECs) were incorporated into EngNT fabricated from collagen hydrogels using the gel aspiration-ejection (GAE) technique and maintained in vitro to allow endothelial network formation. Results: At the mRNA and protein level, pluripotency marker expression decreased and endothelial cell marker expression increased over the course of hiPSC differentiation to endothelial cells. The derived endothelial cells expressed CD31, CD144, ENG, VEGFR2, and VWF, and formed network structures in the matrix tubulogenesis assay. hiPSC-ECs incorporated into EngNT were viable and aligned. They formed highly aligned tube-like structures containing lumens after four days in culture and the EngNT constructs supported neurite growth in vitro when co-cultured with rat dorsal root ganglion (DRG) neurons. Conclusions: This work rapidly generated engineered nerve tissue containing highly aligned endothelial tube-like structures, resembling key features of the early nerve regeneration bridge. Therefore, this 3D engineered tissue provides a platform to study the effects of endothelial cell structures in nerve repair treatment and translational development. Full article

Background: Following myocardial infarction (MI), cardiac fibroblasts (CFs) adopt distinct phenotypes to ensure scar formation and healing. Although leukocytes are a critical driver of post-MI healing, the role of neutrophils in modulating CF phenotype remains insufficiently explored. We therefore investigated the impact of soluble mediators released by neutrophil subtypes found post-MI—pro-inflammatory (N1) and anti-inflammatory (N2)—on shaping CFs phenotype. Methods: In vitro, human 3D grown CFs were indirectly co-cultured with N1 or N2 neutrophil-like cells using a two-chamber Transwell system. After 24 h, expression of inflammatory, remodeling, and pro-fibrotic markers was evaluated in fibroblasts and conditioned media. In vivo, soluble mediators derived from polarized mouse neutrophils (SN1 or SN2) were injected into the infarcted myocardium of C57BL/6J after MI surgery. The effects on the healing process were investigated at 1 and 7 days post-MI. Results: In vitro, CFs were found to exhibit a pro-inflammatory and matrix-degrading phenotype following indirect co-culture with N1 cells, characterized by overexpression of IL-1β, IL-6, MCP-1, and metalloproteases MMP-3/MMP-9. In vivo, both SN1 and SN2 treatments significantly reduced pro-inflammatory markers IL-1β and IL-6 gene expression at day 1 post-MI (inflammatory phase). At day 7 post-MI (resolution phase), SN1/SN2 treatments continued to limit local inflammation, while mitigating fibrotic remodeling by reducing CCN2, α-SMA, and key extracellular matrix proteins. Conclusions: Together, these findings suggest that while N1-derived mediators promote a pro-inflammatory fibroblast phenotype in vitro, factors secreted by both N1 and N2 support a more balanced reparative response in vivo, by limiting local inflammation and potentially mitigating adverse remodeling post-MI. Full article