Search Results (329)

Neuroinflammation is a key feature of Alzheimer’s disease (AD), and stem cell therapies have emerged as promising candidates due to their immunomodulatory properties. Neuro-Cells (NC), a combination of unmodified mesenchymal stem cells (MSCs) and hematopoietic stem cells (HSCs), have demonstrated therapeutic potential in models of central nervous system (CNS) injury and neurodegeneration. Here, we studied the effects of NC in APPswe/PS1dE9 mice, an AD mouse model. Twelve-month-old APPswe/PS1dE9 mice or their wild-type littermates were injected with NC or vehicle into the cisterna magna. Five to six weeks post-injection, cognitive, locomotor, and emotional behaviors were assessed. The brain was stained for amyloid plaque density using Congo red, and for astrogliosis using DAPI and GFAP staining. Gene expression of immune activation markers (Il-1β, Il-6, Cd45, Tnf) and plasticity markers (Tubβ3, Bace1, Trem2, Stat3) was examined in the prefrontal cortex. IL-6 secretion was measured in cultured human monocytes following endotoxin challenge and NC treatment. Untreated APPswe/PS1dE9 mice displayed impaired learning in the conditioned taste aversion test, reduced object exploration, and anxiety-like behavior, which were improved in the NC-treated mutants. NC treatment normalized the expression of several immune and plasticity markers and reduced the density of GFAP-positive cells in the hippocampus and thalamus. NC treatment decreased amyloid plaque density in the hippocampus and thalamus, targeting plaques of <100 μm². Additionally, NC treatment suppressed IL-6 secretion by human monocytes. Thus, NC treatment alleviated behavioral deficits and reduced amyloid plaque formation in APPswe/PS1dE9 mice, likely via anti-inflammatory mechanisms. The reduction in IL-6 production in human monocytes further supports the potential of NC therapy for the treatment of AD. Full article

Background: Alcohol-associated liver disease (ALD) is a primary global health concern, exacerbated by oxidative stress, inflammation, and gut barrier dysfunction. Conventional phytocompounds exhibit hepatoprotective potential but are hindered by low bioavailability. This study aimed to evaluate the hepatoprotective and gut-barrier-restorative effects of green-synthesized silver nanoparticles (AgNPs) derived from Enicostemma littorale, a medicinal plant known for its antioxidant and anti-inflammatory properties. Methods: AgNPs were synthesized using aqueous leaf extract of E. littorale and characterized using UV-Vis, XRD, FTIR, DLS, and SEM. HepG2 (liver) and Caco-2 (colon) cells were exposed to 0.2 M ethanol, AgNPs (1–100 µg/mL), or both, to simulate ethanol-induced toxicity. A range of in vitro assays was performed to assess cell viability, oxidative stress (H₂DCFDA), nuclear and morphological integrity (DAPI and AO/EtBr staining), lipid accumulation (Oil Red O), and gene expression of pro- and anti-inflammatory, antioxidant, and tight-junction markers using RT-qPCR. Results: Ethanol exposure significantly increased ROS, lipid accumulation, and the expression of inflammatory genes, while decreasing antioxidant enzymes and tight-junction proteins. Green AgNPs at lower concentrations (1 and 10 µg/mL) restored cell viability, reduced ROS levels, preserved nuclear morphology, and downregulated CYP2E1 and SREBP expression. Notably, AgNPs improved the expression of Nrf2, HO-1, ZO-1, and IL-10, and reduced TNF-α and IL-6 expression in both cell lines, indicating protective effects on both liver and intestinal cells. Conclusions: Green-synthesized AgNPs from E. littorale exhibit potent hepatoprotective and gut-barrier-restoring effects through antioxidant, anti-inflammatory, and antilipidemic mechanisms. These findings support the therapeutic potential of plant-based nanoparticles in mitigating ethanol-induced gut–liver axis dysfunction. Full article

10X Xenium is an in situ spatial transcriptomics platform that enables single-cell and subcellular-level gene expression analysis. In Xenium data analysis, defining matched keypoints to align H&E and spatial transcriptomic images is critical for cross-referencing sequencing and histology. Currently, it is labor-intensive for domain experts to manually place keypoints to perform image registration in the Xenium Explorer software. We present Xenium-Align, a keypoint identification method that automatically generates keypoint files for image registration in Xenium Explorer. We validated our proposed method on 14 human kidney samples and one human skin Xenium sample representing healthy and diseased states, with expert manually marked results. These results show that Xenium-Align could generate accurate keypoints for automatically implementing image alignment in the Xenium Explorer software for spatial transcriptomics studies. Our future research aims to optimize the method’s runtime efficiency and usability for image alignment applications. Full article

Nowadays, healthcare systems face two global challenges: the rise of multidrug-resistant pathogens and the growing incidence of cancer. Due to their broad spectrum of activities, antimicrobial peptides emerged as potential alternatives against both threats. Our group previously described the antifungal activity of the α-helical peptide Cm-p5, a derivative of the natural peptide Cm-p1, isolated from the coastal mollusk Cenchritis muricatus; however, its anti-cancer properties remained unexplored. Analyses through calorimetry and molecular dynamics simulations suggest the relevance of phosphatidylserine for the attachment of Cm-p5 to cancer cell membranes. Cm-p5 exhibited cytotoxic activity in a dose-dependent manner against A375 melanoma cells, without toxicity against non-malignant cells or hemolytic activity. DAPI/PI and DiSC3(5) staining confirmed permeabilization, disruption, and depolarization of A375 cytoplasmic membranes by Cm-p5. Furthermore, Annexin V-FITC/PI assay revealed the induction of cellular death in melanoma cells, which can result from the cumulative membrane damage and oxidative stress due to the overproduction of reactive oxygen species (ROS). Moreover, after the treatment, the proliferation of A375 cells was dampened for several days, suggesting that Cm-p5 might inhibit the recurrence of melanomas. These findings highlight the multifunctional nature of Cm-p5 and its potential for treating malignant melanoma. Full article

Trans fats are linked to numerous chronic diseases and cellular dysfunction; however, Spain has not implemented effective regulatory measures to restrict their presence in food products. This study addressed these gaps by analyzing trans fat content in commercial pastries sold in Spain and their biological impacts on mesenchymal stem cells, further examining its compliance with international guidelines. Also, a novel and scalable method for extracting fatty acids from pastry samples was developed and applied, enabling precise analysis using gas chromatography alongside sensory property assessments. The findings revealed significant variability in TFA levels across samples. To assess the biological implications of these TFAs, mesenchymal stem cells (MSCs) were cultured to perform dose–response experiments using two selected pastry samples with the highest TFA content. Cellular adhesion, cytotoxicity, and proliferation were evaluated through MTT assays, bright-field, and fluorescence staining using FITC and DAPI markers. Results demonstrated dose-dependent impacts of TFAs on MSC viability, including reduced adhesion and proliferation alongside increased cytotoxicity. This study underlines the need for stricter regulatory frameworks to monitor TFA levels worldwide, including in Spain’s food industry. Additionally, it highlights the potential health risks associated with excessive TFA consumption, particularly concerning cellular health and growth mechanisms, which provide insights into its potential bioaccumulation implications. These findings provide a foundation for further research into dietary guidelines and industrial practices aimed at minimizing TFA exposure while promoting public health safety. Full article

Dirigent proteins (DIRs) are pivotal regulators of lignin/lignan biosynthesis and play multifaceted roles in plant development and stress adaptation. Despite their functional significance, DIR genes remain unexplored in Rosa chinensis, a globally important woody ornamental species. This study identified 33 RcDIRs through whole-genome analysis, including their chromosomal distribution, phylogenetic relationships, collinearity, protein and gene structure, conserved motifs, and cis-acting element distribution, and classified them into three phylogenetically independent subgroups (DIR-a, DIR-b/d, and DIR-e). Notably, the DIR-e subgroup includes an exclusive tandem cluster comprising RcDIR7-RcDIR12, representing the largest lineage-specific RcDIR expansion in R. chinensis. Structural characterization revealed that most RcDIRs exhibit a conserved single-exon architecture. Promoter cis-element analysis uncovered abundant stress-/hormone-responsive elements and three pollen-specific motifs (AAATGA, POLLEN1LELAT52, GTGANTG10), with RcDIR12 from the DIR-e cluster showing high pollen-specific regulatory potential. Experimental validation included cloning the RcDIR12 promoter from R. chinensis ‘Old Blush’, constructing proRcDIR12::GUS vectors, and conducting histochemical GUS assays with pollen viability/DAPI staining in transgenic Arabidopsis. Histochemical assays demonstrated GUS activity localization in mature trinucleate pollen grains, marking the first experimental evidence of pollen-specific DIRs in rose. Our findings not only elucidate the DIR family’s genomic organization and evolutionary innovations in R. chinensis but also establish proRcDIR12 as a molecular tool for manipulating pollen development in plants. Full article

Chitosan–hydroxyapatite (CS-HAp) biocomposites, combining the biocompatibility and bioactivity of chitosan with the osteoconductive properties of hydroxyapatite, are emerging as promising candidates for tissue engineering applications. These materials consistently exhibit excellent cytocompatibility, with cell viability rates greater than 95% in MTT and Neutral Red Uptake assays, and minimal cytotoxicity, as demonstrated by low levels of cell death in DAPI and Trypan blue staining. More importantly, CS-HAp biocomposites modulate the immune environment by enhancing the expression of anti-inflammatory cytokines (IL-10 and IL-4) and the pro-inflammatory cytokine TGF-β, while avoiding significant increases in TNF-α, IL-6, or NF-κB expression in fibroblast cells exposed to HAC and HACF scaffolds. In an in vivo dermatitis model, these biocomposites reduced mast cell counts and plasma histamine levels and significantly decreased pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), JAK1/3, VEGF, and AnxA1 levels. Structurally, HACF scaffolds demonstrated larger average pore sizes (95 µm) compared to HAC scaffolds (74 µm), with porosities of 77.37 ± 2.4% and 65.26 ± 3.1%, respectively. These materials exhibited high swelling ability, equilibrium water content, and controlled degradation over a week in culture media. In addition to their immunomodulatory effects, CS-HAp composites promote essential cellular activities, such as attachment, proliferation, and differentiation, thereby supporting tissue integration and healing. Despite these promising findings, significant gaps remain in understanding the underlying mechanisms of immune modulation by CS-HAp biocomposites, and formulation-dependent variability raises concerns about reproducibility and clinical application. Therefore, a comprehensive review is essential to consolidate existing data, identify key knowledge gaps, and standardize the design of CS/HAp composites for broader clinical use, particularly in immunomodulatory and regenerative medicine contexts. Full article

Critical-sized bone defects or CSDs result from bone loss due to trauma, tumor removal, congenital defects, or degenerative diseases. Though autologous bone transplantation is the current gold standard in treating CSDs, its limitations include donor-site morbidity, unavailability of donor bone tissues, risk of infection, and mismatch between the bone geometry and the defect site. Customized scaffolds fabricated using 3D printing and biocompatible materials can provide mechanical integrity and facilitate osseointegration. Ti-6Al-4V (Ti64) is one of the most widely used commercial alloys in orthopedics. To avoid elastic modulus mismatch between bones and Ti64, it is imperative to use porous lattice structures. Ti64 scaffolds with diamond, cubic, and triply periodic minimal surface (TPMS) gyroid lattice architectures were fabricated using selective laser melting (SLM)with pore sizes ranging from 300 to 900 μm using selective laser melting and evaluated for mechanical and biological performance. Increasing pore size led to higher porosity (up to 90.54%) and reduced mechanical properties. Young’s modulus ranged from 13.18 GPa to 1.01 GPa, while yield stress decreased from 478.16 MPa to 14.86 MPa. Diamond and cubic scaffolds with 300–600 μm pores exhibited stiffness within the cortical bone range, while the 900 μm diamond scaffold approached trabecular stiffness. Gyroid scaffolds (600–900 μm) also showed modulus and yield strength within the cortical bone range but were not suitable for trabecular applications due to their higher stiffness. Cytocompatibility was confirmed through leachate analysis and DAPI-stained osteoblast nuclei. The biological evaluation reported maximum cell adherence in lower pore sizes, with gyroid scaffolds showing a statistically significant (p < 0.01) increase in cell proliferation. These findings suggest that 300–600 μm lattice scaffolds offer an optimal balance between mechanical integrity and biological response for load-bearing bone repair. Full article

Sphaerophoria rueppellii is a Palearctic hoverfly widely used as a native biocontrol agent against aphid pests in Mediterranean agroecosystems. In this study, we present a cytogenetic analysis and characterization of the mitochondrial genome of this species. Chromosomal preparations, obtained from third-instar larvae, were used for conventional staining, DAPI staining and C-banding techniques, and major ribosomal DNA (rDNA) location by fluorescence in situ hybridization (FISH). Karyotype analysis revealed a diploid number of 2n = 10, with heterochromatic blocks in the pericentromeric regions of all autosomes and rDNA clusters on both sex chromosomes. The complete mitochondrial genome (16,605 bp) was sequenced and annotated using next-generation sequencing and assembly pipelines. It contains the typical 37 mitochondrial genes and a highly A + T-rich control region with tandem repeats. Gene order and codon usage were conserved compared with other Syrphidae. Phylogenetic reconstruction based on mitochondrial protein-coding genes clarifies the species’ placement within the Syrphini tribe. Our results contribute valuable genomic and cytogenetic information that supports comparative analyses and may aid in taxonomic clarification within the genus. These findings also offer key data that could guide the genetic optimization of S. rueppellii as an efficient, environmentally safe biological control agent in sustainable agriculture. Full article

In the field of fish genetic breeding, accurately determining the DNA content and ploidy of fish is of great significance. This article introduces the use of flow cytometry (FCM) to measure the DNA content and conduct ploidy analysis by sampling different tissues of freshwater fish species. It describes the FCM detection methods and their effectiveness for different individual tissues. These tissues include embryos and fry, as well as the blood, caudal fins, and sperm of adult live fish, and also specific tissues such as testes, ovaries, gills, spleens, and livers under anatomical conditions. Moreover, the application of ploidy detection to different tissues or individuals in different stages in the practice of fish genetic breeding is analyzed. This research covers samples from different growth stages and a variety of tissue types. The results show that this method exhibits high stability and reliability in the detection of different tissue samples, providing solid data support for subsequent research. It holds significant application value in fish genetic breeding. Full article

This study explores the antioxidant activity and antioxidant mechanism of phenolic compounds (including free (FP), esterified (EP), and bound phenolic (BP)) from Dendrobium fimbriatum Hook (DFH) stems, before and after ultra-high pressure (UHP) treatment. A total of 374 compounds were identified, with 149 showing significant differences in DFH phenolic extracts before and after UHP treatment. UHP treatment significantly increased the total phenolic content (TPC) and total flavonoid content (TFC) and enhanced antioxidant activity in vitro. Particularly, the UEP-DFH, IC₅₀ values in ABTS and DPPH were reduced by 49.6% and 64.1%, respectively. In H₂O₂-treated HepG2 cells, the extracts demonstrated significant cytoprotective effects, including increased cell viability, ROS reduction, and enhanced GSH levels by 17.8% (UFP-DFH) and 12.5% (UEP-DFH). The activities of GS, GCL, GR, GSH-Px, SOD, CAT, NQO1, and HO-1 were also elevated in UHP-treated extracts. DAPI staining indicated that the extracts promoted nuclear Nrf2 expression, particularly UFP-DFH and UEP-DFH. Molecular docking indicated that vanillic acid could competitively bind to the Keap1-Kelch domain, facilitating activation of the antioxidant pathway. Overall, UHP treatment enhanced both extraction efficiency and antioxidant activity, making it a promising method for improving the bioactivity of DFH in the food and functional food industries. Full article

Purpose: This study aimed to compare the histological characteristics of bone tissue following drilling with three implant systems under different rotational speeds and cooling conditions. Methods: A total of 54 implant bed preparations were performed in four swine ribs using three implant systems: Hiossen ET (Hiossen, Fairfield, NJ, USA), Paltop (Burlington, MA, USA), and Anyridge (Megagen, Daegu, Republic of Korea). Drilling was performed at three speeds (800, 1200, and 1500 rpm) under three cooling conditions: saline at room temperature, saline cooled to 4 °C, and no cooling. Histological evaluation was conducted using a Nikon Eclipse 80i fluorescence microscope (Nikon, Tokyo, Japan) with DAPI and rhodamine staining. Observations were performed at 40× magnification, focusing on the osteotomy wall and surrounding tissue. The samples were assessed based on surface smoothness, compressed tissue presence, carbonization, and adjacent tissue damage. Statistical analysis was performed using the Kruskal-Wallis test with Dunn’s post hoc comparisons to evaluate differences among experimental conditions. Results: The results demonstrated that the Hiossen ET system achieved optimal bone bed quality at 1200 rpm with saline cooling at 4 °C, producing the smoothest osteotomy walls and minimal thermal damage (p = 0.003). The Paltop system performed best at 800 rpm with 4 °C cooling, showing reduced tissue compression and fewer microcracks (p = 0.012). The Anyridge system exhibited the most favorable outcomes at 1200 rpm with saline cooling at room temperature, minimizing soft tissue remnants and preserving bone integrity (p = 0.021). Across all systems, the absence of cooling significantly increased thermal damage, carbonization, and tissue fragmentation, particularly at 1500 rpm (p < 0.001). Conclusions: The use of lower rotational speeds with effective cooling minimized tissue trauma and improved bone bed integrity. Further clinical validation is necessary to confirm the applicability of these results in human bone. Full article

Background/Objectives: Meniscus injuries represent a critical challenge in orthopedic medicine due to the limited self-healing capacity of the tissue. This study presents the development and characterization of polycaprolactone/graphene oxide (PCL/GO) scaffolds fabricated using 3D bioprinting technology for meniscus cartilage regeneration. Methods: GO was incorporated at varying concentrations (1%, 3%, 5% w/w) to enhance the bioactivity, mechanical, thermal, and rheological properties of PCL scaffolds. Results: Rheological analyses revealed that GO significantly improved the storage modulus (G’) from 36.1 Pa to 97.1 Pa and the yield shear stress from 97.2 Pa to 507.1 Pa, demonstrating enhanced elasticity and flow resistance. Mechanical testing showed that scaffolds with 1% GO achieved an optimal balance, with an elastic modulus of 614 MPa and ultimate tensile strength of 46.3 MPa, closely mimicking the native meniscus’s mechanical behavior. FTIR analysis confirmed the successful integration of GO into the PCL matrix without disrupting its chemical integrity, while DSC analysis indicated improved thermal stability, with increases in melting temperatures. SEM analysis demonstrated a roughened surface morphology conducive to cellular adhesion and proliferation. Fluorescence microscopy using DAPI staining revealed enhanced cell attachment and regular nuclear distribution on PCL/GO scaffolds, particularly at lower GO concentrations. Antibacterial assays exhibited larger inhibition zones against E. coli and S. aureus, while cytotoxicity tests confirmed the biocompatibility of the PCL/GO scaffolds with fibroblast cells. Conclusions: This study highlights the potential of PCL/GO 3D-printed scaffolds as biofunctional platforms for meniscus tissue engineering, combining favorable mechanical, rheological, biological, and antibacterial properties. Full article

Background: The standard diagnostic approach for prostate cancer (PCa) diagnosis consists of serum prostate-specific antigen (PSA) testing, digital rectal examination (DRE) and image-guided targeted biopsies. Given the invasive nature, potential adverse events and costs associated with these techniques, alternative approaches have been investigated, specifically with serum and urine assays. The work presented here is intended to further validate a novel noninvasive optical technique for PCa detection, targeting the VPAC genomic receptors that are overexpressed on prostate cancer’s malignant cells (MC), in non-DRE voided urine. Methods: Patients (N = 62) who had image-guided biopsy and histologically confirmed localized PCa, and who were scheduled for radical prostatectomy, provided a non-DRE voided urine sample prior to surgery. Urine was cytocentrifuged and cells fixed on a glass slide, incubated with 0.5 μg TP4303 (a receptor-specific fluorophore developed in our laboratory with high affinity for VPAC), excess washed and treated with 4,6-diamidodino-2-phenylindole (DAPI) for nuclear staining. The field of cells on each slide was analyzed using a Zeiss AX10 Observer microscope (20×). The total number of cells and MC were then counted, and the florescent intensity around each MC was measured using Zeiss software. Additionally, non-DRE voided urine samples collected from clinically determined BPH patients (N = 97), were also analyzed similarly. Results: Urine samples from 62 patients were processed and analyzed. Mean PSA levels by Gleason grade (GG) group were 6.5 ± 4.1 ng/mL for GG1 (N = 10), 7.2 ± 3.8 for GG2 (N = 31), 13.2 ± 14.6 for GG3 (N = 13), 6.2 ± 2.2 for GG4 (N = 2) and 50.2 ± 104.9 for GG5 (N = 6). Like the PSA, % MC shed (66.7 ± 27.7) in voided urine and the fluorescent intensity (35.8 ± 5.7) were highest in patients with GG5 prostate cancer. All PCa patients in GG1 to GG5 shed MC in voided urine with increasing % of MC and increasing fluorescence intensity which correlated with the increasing GG for PCa. For BPH, the specificity for the assay was 89.6% (95% CI:81.9–94.9%), PPV was 0.0% and NPV was 100% (95.9% CI, 95.9–100%). Conclusions: These data indicate the following: (i) PCa MC shed in non-DRE voided urine can be detected by targeting VPAC receptors, (ii) MC are shed in non-DRE voided urine with increasing quantity, corresponding to the severity of the disease, and (iii) this non-DRE voided urine optical assay provides a simple, noninvasive, and reliable method for the preliminary detection of PCa with potentially a lower cost than the currently available pre-biopsy detection technologies. Full article

Background/Objectives: Paspalum notatum is a key multipurpose species native to American grasslands. This study provides, for the first time, a detailed karyotype analysis of diploid (2n = 2x = 20) and tetraploid (2n = 4x = 40) accessions of P. notatum, the most common cytotypes within the species. Methods: The constitutive heterochromatin patterns revealed using CMA-DA-DAPI staining and genome size estimations are novel contributions to the understanding of the N genome in Paspalum. Results: Chromosomes were small (1.1–2.3 µm), with the diploid karyotype comprising nine metacentric pairs (one bearing microsatellites on the short arms, pair 6) and one submetacentric pair. In tetraploids, the diploid karyotype was duplicated. Heterochromatin analysis revealed two CMA⁺⁺/DAPI⁻ bands located on the short arm and satellite of chromosome 6 in diploids, while tetraploids exhibited two to three CMA⁺⁺/DAPI⁻ and one to two CMA⁺⁺/DAPI⁰ bands. The proportion of GC-rich heterochromatin represented 2.8 and 3.47% of the total chromosome length in diploid and tetraploid cytotypes, respectively. Genome size analysis revealed a reduction in monoploid genome size in tetraploids (1Cx = 0.678 pg) compared to diploids (1Cx = 0.71 pg), consistent with the autopolyploid origin hypothesis. Conclusions: These findings provide essential cytogenetic insights and suggest only minor structural changes in the N genome following polyploidization, which could guide future studies integrating genomic and cytogenetic maps of P. notatum. Full article