Search Results (341)

Leishmaniasis is an infectious disease common in tropical and subtropical regions worldwide. This study aimed to develop a topical meglumine antimoniate gel (MA-gel) for the treatment of cutaneous leishmaniasis. The MA-gel was characterized in terms of morphology, pH, swelling, porosity, rheology, and thermal properties by differential scanning calorimetry (DSC). Biopharmaceutical evaluation included in vitro drug release and ex vivo skin permeation. Safety was evaluated through biomechanical skin property measurements and cytotoxicity in HaCaT and RAW 267 cells. Leishmanicidal activity was tested against promastigotes and amastigotes of Leishmania infantum, and in silico studies were conducted to explore possible mechanisms of action. The composition of the MA-gel included 30% MA, 20% Pluronic^® F127 (P407), and 50% water. Scanning electron microscopy revealed a sponge-like and porous internal structure of the MA-gel. This formula exhibited a pH of 5.45, swelling at approximately 12 min, and a porosity of 85.07%. The DSC showed that there was no incompatibility between MA and P407. Drug release followed a first-order kinetic profile, with 22.11 µg/g/cm² of the drug retained in the skin and no permeation into the receptor compartment. The MA-gel showed no microbial growth, no cytotoxicity in keratinocytes, and no skin damage. The IC₅₀ for promastigotes and amastigotes of L. infantum were 3.56 and 23.11 µg/mL, respectively. In silico studies suggested that MA could act on three potential therapeutic targets according to its binding mode. The MA-gel demonstrated promising physicochemical, safety, and antiparasitic properties, supporting its potential as a topical treatment for cutaneous leishmaniasis. Full article

Humulus lupulus L. (hop) is a multipurpose crop valued for its essential role in beer production and for its bioactive compounds with recognized medicinal properties. Otherwise, climate change represents a major challenge to agriculture, particularly impacting the cultivation of crops with stenoecious characteristics, such as hop. This highlights the urgent need to enhance crop resilience to adverse environmental conditions. The phytohormone abscisic acid (ABA) is a key regulator of plant responses to abiotic stress, yet the ABA signaling pathway remains poorly characterized in hop. Harnessing the publicly available hop genomics resources, we identified eight members of the PYRABACTIN RESISTANCE 1 LIKE ABA receptor family (HlPYLs). Phylogenetic and gene structure analyses classified these HlPYLs into the three canonical ABA receptor subfamilies. Furthermore, all eight HlPYLs are likely functional, as suggested by the protein sequence visual analysis. Expression profiling indicates that ABA perception in hop is primarily mediated by the HlPYL1-like and HlPYL8-like subfamilies, while the HlPYL4-like group appears to play a more limited role. Structure modeling and topology predictions of HlPYL1b and HlPYL2 provided insights into their potential functional mechanisms. To assess the physiological relevance of ABA signaling in hop, we evaluated the impact of exogenous ABA application during the ex vitro acclimatization phase. ABA-treated plants exhibited more robust growth, reduced stress symptoms, and improved acclimatization success. These effects were associated with reduced leaf transpiration and enhanced stomatal closure, consistent with ABA-mediated drought tolerance mechanisms. Altogether, this study provides the first comprehensive characterization of ABA receptor components in hop and demonstrates the practical utility of ABA in improving plant performance under ex vitro conditions. These findings lay the groundwork for further functional studies and highlight ABA signaling as a promising target for enhancing stress resilience in hop, with broader implications for sustainable agriculture in the face of climate change. Full article

An efficient in vitro propagation protocol for Eulophia bicallosa was developed using asymbiotic seed germination and protocorm proliferation. The effect of light on seed germination and development was evaluated on Vacin and Went (VW) medium under five conditions: darkness, white, green, red, and blue light for 24 weeks. Blue and red light significantly accelerated seed development, allowing progression to stage 5 within 24 weeks. For protocorm proliferation, six semi-solid culture media were tested. Half-strength Murashige and Skoog (½MS) medium yielded the best results after 8 weeks, producing the highest numbers of shoots (1.0), leaves (1.1), and roots (4.2) per protocorm, with 100% survival. The effects of organic additives were also evaluated using coconut water and potato extract. A combination of 200 mL L⁻¹ coconut water and 50 g L⁻¹ potato extract enhanced shoot formation (1.7 shoots), while 150 mL L⁻¹ coconut water with 50 g L⁻¹ potato extract increased both leaf (1.9) and root (8.8) numbers. The effects of cytokinins (benzyladenine (BA), kinetin (6-furfurylaminopurine), and thidiazuron (TDZ)) and auxins (indole-3-acetic acid (IAA), α-naphthalene acetic acid (NAA), indole-3-butyric acid (IBA), and 2,4-dichlorophenoxyacetic acid (2,4-D)) were investigated using ½MS medium supplemented with each plant growth regulator individually at concentrations of 0, 0.1, 0.5, 1.0, and 2.0 mg L⁻¹. Among the cytokinins, 0.1 mg L⁻¹ BA produced the highest survival rate (96%), while 1.0 mg L⁻¹ BA induced the greatest shoot formation (93%, 2.3 shoots). Among the auxins, 0.1 mg L⁻¹ IAA resulted in the highest survival (96%), and 1.0 mg L⁻¹ IAA significantly enhanced root induction (4.2 roots per protocorm). Acclimatization in pots containing a 1:1:1 (v/v) mixture of pumice, sand, and soil resulted in 100% survival. This protocol provides a reliable and effective approach for the mass propagation and ex situ conservation of E. bicallosa. Full article

Essential oils (EOs) are widely recognized for their antifungal properties, but their efficacy against specific phytopathogenic fungi associated with banana (Musa paradisiaca) rot remains underexplored. This study aimed to evaluate the antifungal potential of EOs from Origanum vulgare, Salvia rosmarinus, Syzygium aromaticum, Thymus vulgaris, Cinnamomum verum, and Ocimum basilicum against five fungal species isolated from infected banana peels. Fungal isolates were obtained using PDA medium supplemented with chloramphenicol and were purified by weekly subculturing. Morphological and microscopic characterization was complemented by molecular identification based on ITS sequencing and phylogenetic reconstruction using Neighbor-Joining and UPGMA methods in MEGA v11. In vitro and ex vivo antifungal assays were performed at EO concentrations ranging from 200 to 1000 ppm. Thyme oil exhibited the strongest inhibitory effect, with complete growth suppression at 1000 ppm. Cinnamon and oregano also demonstrated effective inhibition at 600 ppm, while clove, rosemary, and basil were markedly less effective. Statistical analysis confirmed significant effects of EO type and concentration on fungal growth (p < 0.001). Molecular results showed strong phylogenetic support for isolate identification, with bootstrap values above 93% in most clades. These findings support the selective use of specific EOs as sustainable alternatives to synthetic fungicides in the postharvest management of banana diseases and provide a molecularly supported basis for their targeted application in integrated control strategies. Full article

Background/Objectives: Radiolabeled biomolecules specifically targeting overexpressed structures on tumor cells hold great potential for prostate cancer (PCa) imaging and therapy. Due to heterogeneous target expression, single radiopharmaceuticals may not detect or treat all lesions, while simultaneously applying two or more radiotracers potentially improves staging, stratification, and therapy of cancer patients. This study explores a dual-tracer SPECT approach using [¹¹¹In]In-RM2 (targeting the gastrin-releasing peptide receptor, GRPR) and [^99mTc]Tc-PSMA-I&S (targeting the prostate-specific membrane antigen, PSMA) as a proof of concept. To mimic heterogeneous tumor lesions in the same individual, we aimed to establish a dual xenograft mouse model for preclinical evaluation. Methods: CHO-K1 cells underwent lentiviral transduction for human GRPR or human PSMA overexpression. Six-to-eight-week-old female immunodeficient mice (NOD SCID) were subsequently inoculated with transduced CHO-K1 cells in both flanks, enabling a dual xenograft with similar target density and growth of both xenografts. Respective dual-isotope imaging and γ-counting protocols were established. Target expression was analyzed ex vivo by Western blotting. Results: In vitro studies showed similar target-specific binding and internalization of [¹¹¹In]In-RM2 and [^99mTc]Tc-PSMA-I&S in transduced CHO-K1 cells compared to reference lines PC-3 and LNCaP. However, in vivo imaging showed negligible tumor uptake in xenografts of the transduced cell lines. Ex vivo analysis indicated a loss of the respective biomarkers in the xenografts. Conclusions: Although the technical feasibility of a dual-tracer SPECT imaging approach using ¹¹¹In and ^99mTc has been demonstrated, the potential of [^99mTc]Tc-PSMA-I&S and [¹¹¹In]In-RM2 in a dual-tracer cocktail to improve PCa diagnosis could not be verified. The animal model, and in particular the transduced cell lines developed exclusively for this project, proved to be unsuitable for this purpose. The in/ex vivo experiments indicated that results from an in vitro model may not necessarily be successfully transferred to an in vivo setting. To assess the potential of this dual-tracer concept to improve PCa diagnosis, optimized in vivo models are needed. Nevertheless, our strategies address key challenges in dual-tracer applications, aiming to optimize future SPECT imaging approaches. Full article

Anthracnose is a disease caused by phytopathogenic fungi such as Colletotrichum siamense that attacks plants and fruits causing great postharvest losses. Different alternatives for the control of this fungus have been studied. In the present study, we evaluated the in vitro antifungal activity of the methanolic extracts of Baccharis glutinosa (ExB) and Jacquinia macrocarpa (ExJ) individually, as well as in combination with chitosan (CS), along with their toxicity in different models. Using the radial growth technique, it was observed that the mycelial development of C. siamense was altered and reduced during exposure to the different treatments evaluated during the first hours of incubation, indicating a fungistatic effect. While the cell viability, by colorimetric assay using the XTT salt, showed alteration since the chitosan reduced proliferation by 50%, while the plant extracts and their mixtures with chitosan reduced approximately 40% indicating cell damage, which was confirmed by fluorescence microscopy. In addition, toxicity tests demonstrated that the J. macrocarpa extract significantly affected the germination percentage of Lactuca sativa seeds, whereas radicle length was reduced in all treatments except for chitosan. The larval survival test for Artemia salina with the extracts indicated their potential toxicity by causing up to 60% mortality. The results indicate that ExB and ExJ mixed with CS are a good option for controlling C. siamense; however, at the concentrations used, they exhibit a toxic effect on the evaluated models. Full article

Postharvest fungal infections, particularly by Botrytis cinerea, can cause up to 50% losses in fruits and vegetables, and the overuse of synthetic fungicides has led to resistant pathogen strains. We hypothesized that hexane (Hex) and methanolic (MeOH) extracts from peanut (P) and pecan nut (PN) shells possess antifungal properties effective against B. cinerea in strawberries. To test this, we conducted both in vitro and ex vivo assays using strawberries inoculated with B. cinerea, comparing two controls (T0: water; T1: commercial synthetic fungicide) with four treatments—Hex-P, MeOH-P, Hex-PN, and MeOH-PN—at 1000 and 2000 ppm (in vitro) and 4000 ppm (ex vivo). Total phenolic content (TPC) and antioxidant activity (AA) were also measured. MeOH-P and Hex-PN extracts at 2000 ppm significantly inhibited fungal mycelial growth in vitro. In ex vivo assays, MeOH-P reduced both disease incidence and severity comparably to the synthetic fungicide. MeOH-PN exhibited the highest TPC and AA. These findings support the potential use of MeOH-P extract as a natural alternative to synthetic fungicides for controlling B. cinerea in strawberries during postharvest storage. Full article

Advanced glycation end-products (AGEs) and oxidative stress are recognized as central contributors to the pathogenesis of age-related or diabetic cataracts, diabetic retinopathy (DR), and age-related macular degeneration (AMD). These glycation-related diseases are characterized by impaired redox balance and decreased glutathione (GSH) levels. This review aims to examine the mechanistic links between AGEs and GSH depletion across ocular tissues by integrating in vitro, ex vivo, in vivo, and clinical studies relevant to this topic. The multiple levels of evidence highlight GSH homeostasis as both a biomarker and therapeutic target in glycation-related ocular disorders. Therapeutic strategies aimed at restoring GSH homeostasis under glycation stress are categorized into four mechanistic domains: (I) promoting GSH supply and synthesis, (II) enhancing GSH recycling, (III) mitigating glycation stress, and (IV) reducing oxidative and nitrosative stress. Most of these strategies have been explored via different approaches, and experimental findings with various interventions have shown promise in restoring GSH balance and mitigating AGE-induced damage. A pathological link between GSH depletion and vascular endothelial growth factor (VEGF) overexpression is observed in DR and wet AMD. GSH-centered interventions act upstream to modulate redox homeostasis while anti-VEGF therapies target downstream angiogenesis. This study supports the rationale for a dual-targeting strategy that combines redox-based interventions with VEGF inhibition in glycation-related ocular diseases. Full article

Background: Melanoma is a malignant tumor of melanocytes with an increasing incidence worldwide. Plant-based products are rich in bioactive compounds, offering low toxicity and accessible alternatives for melanoma treatment. A biotechnological approach to obtaining plant-derived produce ensures continuous and high-yield production of medicinally valuable biomass. Objectives: This study aimed to induce and optimize the growth of homogenous callus cultures of Kickxia elatine (L.) Dumort., consequently established a cell suspension culture with a high biomass growth rate, analyzed the phytochemical compositions, and assessed the cytotoxic activity against melanoma cells. Methods/Results: Callus cultures were induced under controlled in vitro conditions on Murashige and Skoog (MS) media supplemented with 2.0 mg L⁻¹ Dicamba and 2.0 mg L⁻¹ 2,4-Dichlorophenoxyacetic acid. The selected callus lines exhibited a high growth index (351.71% ± 27.77) and showed a homogeneous morphology, beige colour, and had friable and watery characteristics. A combination of auxin and cytokinin was found to enhance biomass production significantly. Phytochemical investigations putatively annotated major compounds, including benzoic acid derivatives, phenolic glycosides, phenylpropanoic acids, hydroxycinnamic acid derivatives, and tyrosol derivatives. Methanolic extract (KE-Ex) and 40% methanolic fraction (KE-40Fr) were prepared and tested for cytotoxicity against human fibroblast (MRC-5) and melanoma (MeWo) cell lines using direct cell counting and MTT assay. The crude extract exhibited the strongest cytotoxicity effect on MeWo cells, with IC₅₀ values of 125 ± 8 µg mL⁻¹ after 48 h and 117 ± 7 µg mL⁻¹ after 72 h of treatment. Conclusions: The extract demonstrated a time- and dose-dependent cytotoxic effect, making it a potential candidate for melanoma treatment. Full article

NaCl is the main cause of natural soil salinization. Exploring dark septate endophytes (DSEs) with NaCl tolerance provides information for ecological remediation in saline soil areas. In this study, six DSE strains (Didymella macrostoma (Dm), Paraboeremia selaginellae (Ps), Paraphoma pye (Pp), Paraphoma aquatica (Pa), Acrocalymma ampeli (Aa), and Exophiala xenobiotica (Ex)) isolated from the root sections of Anemone tomentosa were subjected to in vitro NaCl stress experiments and inoculation tests. The results showed that six DSE strains can grow on solid media with different NaCl concentrations (0, 0.2, 0.4, 0.6, 0.8, and 1.0 M) and increase the antioxidant enzyme activities and soluble protein contents to adapt to a salt stress environment. Among these strains, the Pp strain exhibited the greatest biomass accumulation under high NaCl concentrations (1.0 M), indicating greater NaCl tolerance compared to the other five strains. In addition, in the pot experiment, all six DSE strains were able to successfully establish a symbiotic relationship with A. tomentosa, and the Pp strain also showed significant growth-promoting effects on seedlings. In summary, the Pp strain is identified as having strong NaCl tolerance and a significant growth-promoting impact, indicating that it has potential applications as a NaCl-tolerant microbial agent and can be used for bioremediation in saline soils. This research contributes to the basic material and theoretical basis for joint plant–microbe combined remediation in areas prone to soil salinization. Full article

Background: Barrier membranes (BMs) have been used in dental surgical procedures for decades, but their exposure can increase the risk of infections and compromise healing from regenerative procedures. Liquid-leukocyte platelet-rich fibrin (LPRF) products have shown antimicrobial effects and enhance wound healing. This in vitro study aimed to evaluate the antimicrobial effects and cellular responses of LPRF products as adjunctive treatments for barrier membranes, hypothesizing that the two liquid LPRF products could improve antibacterial activity against selected oral pathogen species and augment human gingival fibroblast cellular proliferation on BM. Methods: LPRF exudate (LPRF-EX) and liquid fibrinogen (PLyf), human LPRF products, were prepared with recommended centrifugation protocols and used to treat resorbable (Bio-gide^®) and non-resorbable (Cyto-plast™) BMs. Human gingival fibroblasts (HGFs) were cultured on the treated and untreated BMs. Scanning electron microscopy (SEM) was applied to observe cell adhesion, and CCK-8 assays were used to study cell proliferation. Oral P. gingivalis and A. naeslundii were incubated with the BMs. Bacterial adhesion was visualized using SEM, and colony-forming unit (CFU) counts were obtained. Results: SEM images showed markedly greater fibrin network formation after 7 days on resorbable BM (Bio-gide^®) treated with PLyF, but with no notable differences in other resorbable BM or non-resorbable BM groups with both treatments. CCK-8 assays showed non-significant effects on HGF proliferation at 3 and 5 days. SEM showed A. naeslundii growth inhibition in the LPRF-EX- and PLyf-treated BMs, and the greatest reduction in CFU counts of both P. gingivalis and A. naeslundii was noted with treated Cytoplast™. Conclusions: Within the limitations of this preliminary study, it can be concluded that the LPRF-EX and PLyf treatment of BM induced an antimicrobial effect. Their effects on cellular response were unclear due to the lack of significant findings on SEM analysis. Full article

Oil palm (Elaeis guineensis Jacq.) is the leading global oil-producing crop due to its high oil yield. Increasing global demands for palm oil require efficient propagation. Conventional breeding is practical but slow, making micropropagation an attractive alternative for rapidly multiplying superior genotypes. However, transitioning from in vitro to ex vitro conditions causes physiological stress, restricting survival and productivity. This study assessed gas exchange and chlorophyll fluorescence dynamics during acclimatization from in vitro conditions to field establishment, comparing the seedlings obtained in vitro with conventional seed-derived palm seedlings to conventional seed-derived palms. A pronounced photosynthetic efficiency decline occurred after transfer from in vitro culture, followed by a gradual recovery. The photosynthetic rate (A) increased from 0.86 µmol m⁻² s⁻¹ early in acclimatization to 15.43 µmol m⁻² s⁻¹ in field-established seedlings. Physiological characterization using CO₂ and light response curves identified the reductions in carboxylation efficiency and overall quantum yield CO₂. These biochemical constraints gradually diminished during acclimatization, facilitating a transition from heterotrophic to autotrophic growth. Chlorophyll fluorescence analysis revealed remarkable photoinhibition during initial ex vitro stages, indicated by a decreased maximum quantum efficiency of photosystem II. However, the seedlings progressively restored photochemical function throughout subsequent acclimatization phases. These findings highlight the importance of carefully regulating environmental parameters—particularly irradiance, humidity, and carbon availability—during early seedling acclimatization. The effective management of growth conditions significantly mitigates physiological stress, ensuring robust photosynthetic activity and optimized stomatal regulation. The improved acclimatization practices, therefore, can substantially enhance seedling survival rates, physiological resilience, and the overall field performance of micropropagated oil palms. Future research should focus on refining acclimatization protocols, emphasizing targeted physiological interventions to maximize the efficiency, commercial viability, and sustainability of oil palm clonal propagation. Full article

Background: In tissue engineering, developing injectable hydrogels with tailored mechanical and bioactive properties remains a challenge. This study introduces an injectable hydrogel composite for soft tissue regeneration, composed of oxidized alginate (OA) and N-succinyl chitosan (NSC) cross-linked via Schiff base reaction, reinforced with graphene oxide (GOx) and cyclic arginylglycylaspartic acid (c-RGD). The objective was to create a multifunctional platform combining injectability, bioactivity, and structural stability. Methods: The OA/NSC/GOx-cRGD hydrogel was synthesized through Schiff base cross-linking (aldehyde-amine reaction). Characterization included FTIR (C=N bond at 1650 cm⁻¹), Raman spectroscopy (D/G bands at 1338/1567 cm⁻¹), SEM (porous microstructure), and rheological analysis (shear-thinning behavior). In vitro assays assessed fibroblast viability (MTT) and macrophage TNF-α secretion (ELISA), while ex-vivo injectability and retention were evaluated using chicken cardiac tissue. Results: The hydrogel exhibited shear-thinning behavior (viscosity: 10 to <1 Pa·s) and elastic-dominated mechanics (G′ > G″), ensuring injectability. SEM revealed an interconnected porous structure mimicking native extracellular matrix. Fibroblast viability remained ≥95%, and TNF-α secretion in macrophages decreased by 80% (30 vs. 150 pg/μL in controls), demonstrating biocompatibility and anti-inflammatory effects. The hydrogel adhered stably to cardiac tissue without leakage. Conclusions: The OA/NSC/GOx-cRGD composite integrates injectability, bioactivity, and structural stability, offering a promising scaffold for tissue regeneration. Its modular design allows further functionalization with peptides or growth factors. Future work will focus on translational applications, including scalability and optimization for dynamic biological environments. Full article

The global landscape is marked by climatic, socioeconomic, and demographic complexities, and enhancing food production through byproducts has emerged as a powerful strategy to address these challenges. This review aimed to analyze the potential impacts of VIUSID^® agro, an amino acid–based growth promoter, on plant growth, productivity, and tolerance to salt stress. A quantitative systematic review was conducted utilizing databases such as Web of Science, Scopus, and Google Scholar. The research papers highlighted the positive effects of VIUSID^® agro on growth and production, as well as on physiological and biochemical indices related to salt stress across various crops. Furthermore, this biostimulant can be administered in different doses and through various application methods. The review also examined its effects during the ex vitro acclimatization phase and in tissue culture. The results demonstrated enhanced crop growth, increased biomass accumulation, modulation of photosynthesis, improved enzymatic antioxidant defenses, and maintenance of ionic homeostasis, all contributing to superior crop performance. These findings suggest that VIUSID^® agro is beneficial for a wide range of crops, with its effectiveness primarily attributed to its rich amino acid composition, which influences and modulates various physiological and biochemical processes within plant cells. Full article

K. pneumoniae is a major cause of bovine mastitis worldwide, making it difficult to control due to its resistance to multiple drugs. L. paraplantarum has been explored as a promising new approach to fighting bovine mastitis. In this study, the probiotic potential and safety of L. paraplantarum SDN1.2, as well as its ex vivo and in vivo anti-inflammatory effects against K. pneumoniae-induced mastitis, were comprehensively investigated using bioinformatics analyses and experimental validation methods. The results revealed that L. paraplantarum SDN1.2 exhibits non-hemolytic activity, is not cytotoxic, lacks virulence genes (e.g., adhesion factors, toxins, and invasion factors) and antibiotic resistance genes (e.g., beta-lactamases and tetracycline resistance genes), as supported by whole-genome sequencing, and significantly inhibits the growth of K. pneumoniae, as evaluated by antimicrobial tests. Following further validation in vitro, L. paraplantarum SDN1.2 demonstrated the capability to inhibit the adhesion and invasion of K. pneumoniae to bMECs. In a mouse model of K. pneumoniae-induced mastitis, L. paraplantarum SDN1.2 reduced the extent of neutrophil infiltration and inflammatory lesions. Furthermore, L. paraplantarum SDN1.2 pretreatment significantly reduced myeloperoxidase (MPO) activity and the expression of inflammatory cytokines (IL-6, IL-1β, and TNF-a) in mouse mammary gland tissue. In K. pneumoniae-infected bMECs, L. paraplantarum SDN1.2 significantly lowered lactate dehydrogenase (LDH) levels and expression of inflammatory cytokines such as IL-6, IL-1β, and TNF-α. The results demonstrated that the newly isolated L. paraplantarum SDN1.2 from bovine sources exhibits promising characteristics as a safe probiotic for the alleviation of bovine mastitis due to its safety profile and anti-inflammatory and antibacterial properties. Full article