Search Results (210)

Glioblastomas (GB) are highly aggressive brain tumors with poor patient prognosis and low survival rates. To identify novel therapeutic targets, the tumor microenvironment (TME) is increasingly examined, with a particular focus on biomechanical changes in the extracellular matrix (ECM) that contribute to GB aggressiveness. In GB, the ECM stiffens, regulating cell behavior through mechanotransduction. Preclinical in vitro and ex vivo studies generally report increased stiffness in GB relative to healthy brain tissue, whereas clinical in vivo measurements often report decreased stiffness. This review examines potential causes for this discrepancy, highlighting both biological and technical factors. Preclinical measurements are frequently performed using atomic force microscopy (AFM), while clinical stiffness is assessed via magnetic resonance elastography (MRE). Differences in methodology, including sample preparation, measurement modalities, and spatial scale, partly explain divergent stiffness values. Biological factors such as necrosis, edema, and physical confinement by the skull, which are preserved only in vivo, also contribute to these differences. To reconcile these findings, future research should employ physiologically relevant in vitro models that better replicate in vivo GB biomechanics, together with high-throughput and accurate animal models. Integrating these approaches may clarify the biomechanical landscape of GB and result in more effective therapeutic strategies. Full article

Rosa canina L. is a medicinal and nutritionally valuable species with increasing industrial demand, yet its conventional propagation is limited by low rooting capacity and high genetic heterogeneity. In this study, a complete and reproducible in vitro micropropagation protocol was established, from explant introduction to plantlet acclimatization. Axillary buds were disinfected and introduced into Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP). Shoot multiplication was achieved using sequential cytokinin treatments, and shoot elongation was promoted by adding liquid MS medium containing activated charcoal (AC). The highest and fastest root induction percentage (up to 75%) was obtained on WPM with 2 mg·L⁻¹ IBA and under a 16 h light/8 h dark photoperiod. Light promoted adventitious root formation depending on the nutrient formulation. Thereafter, shoots developed well-structured root systems in vitro, and plantlets fully survived to ex vitro acclimatization. This protocol provides an efficient platform for the large-scale propagation of R. canina and demonstrates that its auxin-driven adventitious rooting is strongly conditioned by the interaction between basal medium composition and photoperiod. Full article

To overcome the seasonal constraints of explant availability and facilitate genetic improvement in Catalpa ovata, this study established a dual-pathway in vitro regeneration system (encompassing adventitious shoot organogenesis and somatic embryogenesis) using mature zygotic embryos. We systematically evaluated the synergistic effects of maternal genotypes, plant growth regulators (PGRs), basal media, and the histone deacetylase inhibitor Trichostatin A (TSA). Genotype screening revealed significant divergence in regenerative potential, with the half-sib family 32F17 exhibiting superior responsiveness (84.7% callus induction). A high cytokinin-to-auxin ratio (ZA3 medium) optimally drove direct shoot organogenesis. For adventitious shoot proliferation, the addition of TDZ significantly improved the multiplication coefficient (up to 2.99 on ZB4 medium), although a physiological trade-off with shoot elongation was observed. In parallel, the application of 10 µM TSA significantly enhanced somatic embryogenesis from embryogenic calli, effectively alleviating the inhibitory constraints of exogenous PGRs. For rhizogenesis, the DKW basal medium proved superior to half-strength MS, with the ZE3 treatment (0.1 mg·L⁻¹ NAA + 0.1 mg·L⁻¹ IBA) yielding the highest rooting frequency (69.6%) and robust root architecture. Notably, while somatic embryo conversion remained recalcitrant, plantlets derived exclusively from the adventitious shoot organogenesis pathway were successfully acclimatized ex vitro. These transplanted plantlets exhibited consistently high survival rates (83.1–84.4%) across all tested genotypes, effectively overcoming the initial genotype-dependent recalcitrance. Collectively, this optimized protocol provides a reliable technical platform for the large-scale clonal propagation and biotechnological breeding of C. ovata. Full article

A rare and threatened pleurocarpous amblystegiaceaen dendrothelmatic moss, Anacamptodon splachnoides, was the subject of in vitro establishment and propagation. Tests on growth and rapid propagation were applied with the aim of preparing plant material for outdoor ex situ conservation and reintroduction into nature. This integrative conservation approach provides novel methodologies applicable to conservation programmes and management strategies, while also providing fundamental insights into the species’ biology. It further enables research on the species without harming its native populations, allowing for the exploration of questions raised during this investigation, such as spore biology and sex expression. A. splachnoides acts as both a shelter and signal species for a special microhabitat type, which also supports other rare organisms, and prolonged drought seems to affect reproduction, spore dispersal, and germinability more strongly than gametophore survival. Full article

The provincial-level registered superior cultivar Camellia chekiangoleosa ‘Ganhongyou 1’ boasts superior economic traits coupled with significant ornamental value, driving demand for an efficient propagation system. Consequently, this study aimed to develop a rapid micropropagation protocol by investigating culture conditions using semi-woody nodal segments with axillary buds as explants on Hyponex basal medium supplemented with varying combinations of plant growth regulators. Contamination was effectively minimized to 18% by a combined approach of surface sterilization (75% ethanol, 0.1% HgCl₂, and 20% NaClO) and incorporating 1 mL/L bactericide into the induction medium. For bud induction, the optimal medium was 2 g/L Hyponex supplemented with 1.0 mg/L 6-benzylaminopurine (6-BA) and 0.2 mg/L indole-3-acetic acid (IAA), achieving an 86.67% induction rate. The best proliferation was achieved on the medium containing 2 g/L Hyponex, 1.0 mg/L 6-BA, 0.15 mg/L 3-indolebutyric acid (IBA), and 0.5 mg/L gibberellic acid (GA₃), yielding a proliferation coefficient of 6.53. A combined strategy, integrating in vitro pre-culture with ex vitro treatment, proved most effective for rooting and acclimatization: shoots were first pre-cultured for 20 days on 1/2 strength Hyponex medium supplemented with 0.5 mg/L 1-naphthaleneacetic acid (NAA) and 2.0 mg/L IBA, followed by ex vitro base treatment with 1.0 g/L ABT (a rooting powder complex) solution before transplantation into seedling bags. This approach resulted in an 88% survival rate. Furthermore, anatomical analysis revealed the origin of adventitious root primordia from phloem parenchyma cells, thereby confirming a phloem-rooting pattern for this species. In conclusion, this study establishes a practical and efficient micropropagation protocol for ‘Ganhongyou 1’, providing a reliable technical foundation for its commercial-scale seedling production. Full article

Background/Objectives: Human milk is a complex biological fluid containing not only macro- and micronutrients but also diverse bioactive molecules, including extracellular RNAs. Although RNA has been detected in milk for decades, only a subset of RNA species has been characterized in detail, and abundant families such as tRNA-, yRNA-, and rRNA-derived fragments remain underexplored. This study aimed to define the composition, fragmentation patterns, stability, and exploratory functional activity of these highly abundant RNAs in human milk. Methods: We performed small RNA sequencing on skim milk samples and analyzed the resulting profiles in comparison with publicly available milk and biofluid datasets. RNA stability assays, Northern blotting, and RT-qPCR were conducted to validate RNA abundance and degradation kinetics. Extracellular vesicles (EVs) and non-vesicular fractions were analyzed to determine the subcellular distribution of RNA species. Exploratory functional assays using synthetic RNA fragments were carried out to assess their ability to modulate cellular responses in vitro. Results: Human milk was found to be highly enriched in small RNA fragments derived from tRNA, yRNA, and rRNA, dominated by a limited set of discrete sequences. These profiles were highly reproducible across independent datasets and distinct biofluids. Orthologal validation assays confirmed their abundance and stability, with RNA levels exceeding those of serum by over two orders of magnitude. Full-length transcripts were enriched in EVs, whereas shorter fragments predominated in the non-vesicular fraction. Synthetic milk-derived exRNAs showed detectable pro-survival activity under stress conditions in vitro. Conclusions: This study reveals that human milk carries a limited set of highly abundant stable sRNA molecules, primarily derived from tRNAs, yRNAs, and rRNAs. These findings provide new insights into the RNA cargo of human milk and offer preliminary evidence that selected sRNA fragments can modulate cellular stress responses in in vitro models. Full article

Background: Serum-free culture of red blood cells (RBCs) typically leads to rapid loss of viability, limiting experimental and translational applications. Lipid-rich formulations and essential oils may provide biocompatible support for RBC integrity while limiting microbial overgrowth. Methods: RBCs from nine healthy adult donors were cultured in serum-free RPMI under four conditions: control, vehicle (olive oil, 1:100 v/v), genuine adenosine triphosphate (ATP)-oil^® (1:100 v/v), and laboratory oil, “mimicking” ATP-oil^®. Cultures were maintained for 18 days. Viability was assessed by light microscopy and trypan blue exclusion; bacterial contamination was qualitatively observed on day 18. Results: Genuine ATP-oil^® maintained 35–45% RBC viability at day 18, whereas control and vehicle cultures declined rapidly. The mimicking preparation did not reproduce these effects. ATP-oil^® immersion was associated with a qualitative reduction in bacterial contamination versus control, consistent with a dual action on RBC preservation and microbial suppression under serum-free conditions. Conclusions: Supplementation with ATP-oil^® substantially prolongs RBC survival and limits bacterial overgrowth in vitro, outperforming commonly used serum or plasma supplements on a per-volume basis. These findings suggest potential applications for improving ex vivo handling or storage of blood components and for reducing background contamination in diagnostic microbiology. Further studies with larger cohorts are warranted to reveal underlying mechanisms and to define active constituents in order to standardize production. Full article

The light spectrum and vapor pressure deficit (VPD) are key environmental factors that significantly influence the morphophysiological development and survival of micropropagated woody plants during acclimatization. However, few studies have focused on their interactive effects under ex vitro conditions. This study examined the combined effects of four light spectra (white, blue, red, and red–blue) and two VPD levels (low: 0.2 kPa; high: 1.0 kPa) on growth, photosynthesis pigments, biochemical indices, and leaf temperature of Pistacia spp. ‘UCB1’ plantlets over a 30-day acclimatization period. The results demonstrated that red–blue light under low VPD significantly enhanced plantlet performance across multiple parameters, resulting in the highest leaflet number (79.25 pieces), stem diameter (2.13 mm), leaf dry weight (0.048 g), leaf fresh weight (0.15 g), root length (1.48 cm), and leaf area (103.3 cm²). Furthermore, this treatment markedly increased anthocyanin, total soluble carbohydrate content, and photosynthetic pigments (chlorophyll a, chlorophyll b, and carotenoids). Principal component and correlation analyses identified that red–blue light under low VPD was strongly associated with traits linked to growth and photosynthetic ability, whereas blue and white light under high VPD showed the weakest responses. Entropy-weighted TOPSIS ranked red–blue light under low VPD as the most effective treatment for balanced morpho-physiological functions during acclimatization. These findings highlight the importance of optimizing spectral quality and VPD to enhance autotrophic transition and ex vitro establishment in pistachio plantlets. These findings are important for improving ex vitro survival and large-scale propagation efficiency of micropropagated pistachio plantlets. Full article

Background/Objectives: Uveal melanoma (UM) is the most common intraocular malignancy in adults. Although brachytherapy of the primary tumor provides an approximate 80% five-year survival, with time, nearly half of patients experience predominant liver metastases. It was proposed that malignant cells migrate early and stay dormant as they adapt to the liver microenvironment. We propose that cancer vaccine-mediated activation of UM-targeted immunity in primary UM patients could prevent progression of metastatic disease from dormant cells or malignant seeds. Thus, this study explored DNA vaccination as a measure to educate the immune system to recognize the most common UM-associated Q209L tumor driver mutation in GNAQ and GNA11 G-alpha proteins. Methods: Several DNA constructs encoding mutated GNAQ were developed and tested for activation of UM-reactive T cells in HLA-A2/Hd transgenic mice and human T cells ex vivo. Results: Constructs containing immune-enhancing PADRE and VP22-derived epitopes boosted T cell responses against mutant GNAQ, which correlated with reduced experimental lung metastases. Ex vivo dendritic cell-mediated T cell activation with vaccine constructs containing optimized structure produced cytolytic T cells that secreted IFN gamma and killed mutated GNAQ-expressing UM cells in vitro. Conclusions: These findings propose the utility of the fusion DNA vaccines in eliciting T cell immunity against UM cells bearing the Q209L mutation in GNAQ/GNA11 protein to prevent the establishment and progression of metastatic disease. Full article

Subcutaneous mycoses are a heterogeneous group of chronic fungal infections, usually acquired through traumatic inoculation of environmental fungi and particularly severe in immunocompromised and critically ill patients. These infections involve pathogens with marked morphological and physiopathological diversity, resulting in significant diagnostic and therapeutic challenges. Conventional treatment relies on systemic antifungals such as amphotericin B, itraconazole, and other azoles; however, these therapies are often limited by poor tissue penetration, adverse effects, and prolonged treatment regimens, especially in vulnerable patient populations. In this context, nanodrugs have emerged as promising alternatives by improving solubility, stability, bioavailability, and targeted delivery to infection sites. This review conducted a comprehensive literature search in PubMed, SciELO, ScienceDirect, Web of Science, and Scopus, identifying 31 eligible studies that developed and evaluated antifungal nanosystems using in vitro, ex vivo, and/or in vivo models. Quantitative outcomes included minimum inhibitory concentration (MIC), colony-forming units (CFU), inhibition halo diameter, and survival assays. Overall, the evidence indicates that several nanosystems may overcome key pharmacological limitations of conventional antifungals and enhance therapeutic outcomes. Nevertheless, important translational challenges remain, including toxicity, long-term safety, scalability, and regulatory approval, which must be addressed before clinical implementation. Full article

In the present study, we developed an efficient and reproducible protocol for in vitro regeneration and Agrobacterium tumefaciens-mediated genetic transformation of Broussonetia papyrifera (L.) L’Hér. ex Vent. (paper mulberry) using leaf explants from a hybrid genotype. First, we optimized surface sterilization of leaf explants. Treatment with 0.6% (w/v) sodium hypochlorite for 8 min, followed by three rinses with sterile water and blotting on sterile filter paper, yielded a 33.60% explant survival rate and reduced contamination to 35.84%. Second, we refined the co-cultivation step for transformation using A. tumefaciens strain EHA105 carrying pCAMBIA1300-35S-eGFP. Leaf discs were infected for 20 min and co-cultured for 2 days on co-cultivation medium overlaid with sterile filter paper, which limited the overgrowth of A. tumefaciens. After co-cultivation, explants were transferred sequentially to callus induction, shoot induction, shoot multiplication, and rooting media supplemented with 250 mg·L⁻¹ cefotaxime and 200 mg·L⁻¹ Timentin, as well as 5.0 mg·L⁻¹ hygromycin at a concentration that completely suppressed regeneration of non-transformed explants. Meanwhile, after transfer to the callus induction medium, eGFP fluorescence was detected in resistant calli as an initial screening for transformants. The integration and expression of the transgene were further confirmed by PCR and quantitative reverse transcription PCR (qRT-PCR) after the resistant calli developed into plantlets. Collectively, this streamlined protocol provides a practical platform for functional genomics and genetic improvement of B. papyrifera. Full article

Oca (Oxalis tuberosa Mol.) is an Andean crop with high nutritional and cultural value; however, its vegetative propagation makes it challenging to ensure a continuous supply of high-quality planting material. In this study, an efficient and reproducible in vitro propagation protocol was established for the oca genotype OT–001 (Amazonas, Peru), integrating shoot multiplication, rooting, and acclimatization. One-centimeter nodal segments were cultured in MS medium supplemented with 6-benzylaminopurine (BAP) or kinetin (KIN) at increasing concentrations ranging from 0.1 to 2.0 mg L⁻¹. For rooting, one-centimeter shoots were transferred to MS medium supplemented with indole-3-butyric acid (IBA) or 1-naphthaleneacetic acid (NAA) at increasing concentrations ranging from 0.1 to 2.0 mg L⁻¹. The variables evaluated four weeks after treatment initiation were regeneration percentage, rooting percentage, number of shoots per explant, number of roots per explant, number of nodes, and shoot length. The regeneration rate reached 100% with both BAP and KIN treatments; however, shoot proliferation was highest with 1.0 mg L⁻¹ BAP, producing an average of 7.4 shoots per explant compared to 2.3 shoots in the control. Meanwhile, KIN concentrations of 0.2–0.5 mg L⁻¹ promoted the development of longer shoots (up to 31.4 mm). In rooting, although the control achieved 93.3%, auxin supplementation improved root architecture. IBA at 0.1 mg L⁻¹ achieved 100% rooting with the longest roots (23.9 mm), while 2.0 mg L⁻¹ IBA maximized the number of roots (14.2 roots per explant). With NAA, the root systems were dense but shorter. The in vitro-regenerated plantlets exhibited 100% survival after 15 and 30 days of acclimatization in sterile agricultural soil, demonstrating the high quality of the plant material obtained. The protocol enables the production of homogeneous and vigorous plantlets throughout the year and provides a practical foundation for the ex situ conservation of oca germplasm and its commercial propagation. It also establishes the basis for advanced applications such as genetic transformation and gene editing. Full article

Non-invasive brain stimulation (NIBS) techniques—including repetitive transcranial magnetic stimulation (rTMS), theta-burst stimulation (TBS), paired associative stimulation (PAS), transcranial direct current stimulation (tDCS), and transcranial alternating current stimulation (tACS)—have emerged as valuable tools for modulating neural activity and promoting plasticity. Traditionally, their effects have been interpreted within a binary framework of long-term potentiation (LTP)-like and long-term depression (LTD)-like plasticity, largely inferred from changes in motor evoked potentials (MEPs). However, existing models do not fully capture the complexity of the biological processes engaged by these techniques and despite extensive clinical application, the cellular and molecular mechanisms underlying NIBS remain only partially understood. This systematic review, conducted in accordance with the PRISMA 2020 guidelines, synthesizes evidence from in vivo, in vitro, and ex vivo studies to delineate how NIBS influences neurotransmission through intracellular signaling, gene expression, and protein synthesis at the cellular level. Emphasis is placed on the roles of classical synaptic models, grounded in Ca²⁺-dependent glutamatergic signaling and receptor phosphorylation dynamics, as well as broader forms of plasticity involving BDNF–TrkB signaling, epigenetic modifications, neuroimmune and glial interactions, anti-inflammatory pathways, and apoptosis- and survival-related cascades. By integrating findings in humans with those in animal and cellular models, we identify both shared and technique-specific molecular mechanisms underlying NIBS-induced effects, highlighting emerging evidence for multi-pathway, non-binary plasticity mechanisms. Understanding these convergent pathways provides a mechanistic foundation for refining stimulation paradigms and improving their translational relevance for treatment of neurological and psychiatric disorders. Full article

Artemisia ludoviciana Nutt. is an important aromatic plant widely used in traditional Mexican medicine for its therapeutic potential. Its medicinal activity is attributed to a wide range of bioactive compounds, including flavonoids. However, overexploitation, habitat loss, climate change and plant diseases threaten its natural populations and diversity. Although the species reproduces both sexually and asexually, conventional propagation methods are often slow, limited by environmental factors, and susceptible to pests and pathogens. Therefore, this research aimed to establish a micropropagation protocol for A. ludoviciana and to evaluate the changes in its phenolic composition and antioxidant activity while adapting to ex vitro conditions. Full-strength Murashige and Skoog (MS) media supplemented with 0.1 mg L⁻¹ 6-benzylaminopurine resulted in the highest number of shoots (3.30 ± 0.34) and shoot length (3.00 ± 0.12 cm). Moreover, 1/2 MS media supplemented with 0.5 mg L⁻¹ indole-3-acetic acid improved the number (14.45 ± 0.56) and quality of roots. Hardening and acclimatization of plantlets showed 100% survival after 10 weeks. Also, the phenolic composition and antioxidant activity of A. ludoviciana changed in response to stress derived from growth conditions. The results support the sustainable use and rapid propagation of the species, as well as provide the basis for the study of secondary metabolism in the plant. Full article

Chronic low-grade inflammation and oxidative stress induced by the exposome represent key drivers of skin aging and related imperfections. The development of experimental models suitable for studying these metabolic processes is therefore of primary importance for the cosmetic industry. In recent years, the role of specialized pro-resolving mediators (SPMs) in the resolution of inflammation has been highlighted; however, in vitro skin models to investigate them are still lacking. In this work, we developed an ex vivo human skin culture model that allows the quantification of maresin 1 (MaR1) production by measuring its concentration in the conditioned culture medium using an ELISA-based assay. The presence and survival of MaR1-synthesizing immune cells, namely Langerhans cells and leukocytes, were quantified during the first days of culture. The model’s ability to modulate MaR1 production was assessed in response to treatment with its precursor, docosahexaenoic acid (DHA), and with a DHA-rich cosmetic ingredient named Isochrysis Galbana Extract. Results demonstrated that the model produces MaR1 even in the absence of stimulation and responds to treatments with a further increase in MaR1 production. Furthermore, the tissue-to-medium ratio required to obtain MaR1 concentrations suitable for effective ELISA quantification was optimized. This model establishes a reproducible and scalable experimental platform for quantifying SPMs and evaluating DHA-based formulations, supporting both cosmetic research and mechanistic investigations. Full article