Search Results (1,362)

Leishmaniases are infections caused by Leishmania parasites and transmitted through the bite of infected female Phlebotomus (Old World) and Lutzomyia (New World) sandflies. The disease disproportionately affects marginalized communities with limited healthcare access. With no approved human vaccines available, leishmaniasis treatment and prevention depend heavily on chemotherapeutics that face growing drug resistance challenges alongside toxicity concerns. The development of safe, effective and affordable vaccines against human leishmaniasis remains a global health priority for disease control and elimination, mostly in resource-limited settings. This review synthesizes progress in leishmaniasis vaccine platforms including live-attenuated parasites, whole-killed parasites, DNA, protein subunit, peptide-based and chimeric/multiepitope vaccines and their homogenous and heterogenous efficacy. Live-attenuated and whole-parasite vaccines have been accounted to elicit robust cellular immunity but pose safety risks, particularly in immunocompromised hosts. While both second- and third-generation vaccines exemplified by LEISH-F1/F3 polyproteins, elicit strong Th1-biased T cell responses in preclinical models, their efficacy in humans remains limited. However, the highlighted collective efforts are pivotal in steering the rational development of future research using various formulations for multiple management of leishmaniasis through cross-protection. Furthermore, emerging strategies including mRNA platforms, nanoparticle delivery, reverse vaccinology, and immunoinformatics offer promising avenues for accelerating vaccine discovery and advancing the development of novel and effective human vaccines. Full article

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9, a genome-editing technology pioneered in 2012, enables the precise correction of deleterious mutations or disruption of disease-causing genes through targeted double-strand breaks (DSBs), offering potential for treating genetic diseases. However, CRISPR/Cas9 can cause off-target cleavage at non-specific DNA sites, leading to unintended insertions or deletions (indels), which limit its safety and applicability despite ongoing improvements in specificity. Recently, prime editing (PE), an advanced CRISPR-derived technology, has been employed with a Cas9 nickase (Cas9n) fused with a reverse transcriptase and a prime editing guide RNA (pegRNA) to enable precise insertions, deletions, and transversions without inducing DSBs, thus reducing risks of indels and chromosomal aberrations. Furthermore, ongoing optimizations, such as improved pegRNA design and enhanced editing efficiency, have expanded the applications of PE in medical therapeutics, agriculture, and fundamental research. This review summarizes recent advancements in the PE system, including optimized pegRNA designs and enzyme engineering for enhanced efficiency and specificity, alongside novel delivery methods. It also evaluates cutting-edge delivery strategies, such as adeno-associated virus (AAV) vectors, lipid nanoparticles (LNPs) and novel extracellular vesicle (EV)-based systems, and explores PE applications in vitro and in vivo, including disease modeling and therapeutic gene correction. Full article

Sensitive quantitative detection of breast cancer gene synthetic sequences is crucial for related biosensing research. To address the limitations of traditional sensors for detecting ultra-low concentrations, this study developed a novel fiber-optic biosensor by combining nanomaterial sensitization with nanoparticle signal amplification strategies. A fiber optic sensor based on single-mode fiber-thin-core fiber-multimode fiber-single-mode fiber structure was fabricated and functionalized with black phosphorus (BP) nano-interface. The Au@cDNA complex was prepared by covalently immobilizing sulfhydryl-modified complementary DNA (cDNA) on the surface of gold nanoparticles (AuNPs). The complex specifically hybridized with the probe DNA (pDNA) immobilized on the surface of the sensor. The experimental results show that this sensor has a sensitivity of 0.793 nm/lgM and a detection limit of 20.27 fM in the concentration range of 100 fM to 100 nM. Specifically, the BP-functionalized sensor exhibits superior dynamic range, higher sensitivity, and lower detection limits for detecting Au@cDNA. The synergistic effect of interfacial sensitization by BP and signal amplification by AuNPs significantly enhances detection performance, providing a promising platform for ultra-sensitive biosensing applications. Full article

Nano-graphene oxide (NGO) is a nanomaterial that has been frequently used in the fields of health and bioengineering in recent years. However, its potential use in semen cryopreservation is still in the exploratory phase. In this study, Angora bucks, a breed with low resistance to cold shock, were used. Sperm was collected from five different Angora bucks, pooled, diluted with a Tris-based egg yolk diluent, and frozen with the addition of NGO at two different sizes (50 and 500 nm) and doses (10 and 50 µg/mL). Nanoparticle characterization was performed using field emission scanning electron microscopy (FE-SEM), dynamic light scattering (DLS), and Fourier-transform infrared spectroscopy (FTIR). Post-thaw sperm analyses were evaluated based on motility and kinematic parameters, mitochondrial membrane potential (MMP), plasma membrane and acrosome integrity (PMAI), and DNA fragmentation. Applying 50 nm NGO at a dose of 50 µg/mL led to statistically significant improvements in motility and PMAI (p < 0.05). The same dose of 500 nm NGO, however, only showed a statistically significant improvement in the PMAI parameter (p < 0.05). No significant differences were observed between the groups for MMP and kinematic parameters (p > 0.05). Conversely, it was found that all sizes and doses of NGO significantly protected post-thaw sperm regarding DNA integrity (p < 0.05). These findings indicate that the NGO, at a size of 50 nm and a dose of 50 µg/mL, improves the post-thaw quality of Angora buck sperm and provides a cryoprotective effect that depends on size and dose. This study provides preliminary data on the potential effects of NGO; however, comprehensive mechanistic and in vivo validation studies are required to establish the biological and clinical validity of these findings. Full article

Monitoring the concentration of doxorubicin (DOX) was critical for tumor treatment, but existing methods failed to cross cell membrane. Here, an electrochemical platform for intracellular DOX detection in MCF-7 cells based on membrane-permeation strategy was developed. A modified gold electrode was prepared via electrodepositing AuNPs and assembling SH-DNA. Concurrently, the silica nanosphere/gold nanocluster-circular transmembrane peptide (SiO₂/AuNCs-iRGD) composite nanoparticles with membrane permeability, tumor targeting, and imaging capability were synthesized. After co-incubation of SiO₂/AuNCs-iRGD with MCF-7 cells and DOX, followed by co-incubation with the DNA-modified electrode, intracellular DOX intercalated into the DNA backbone, and redox-generated electrons were transferred to the electrode to produce a concentration-correlated electrochemical signal. The modification of the electrode, the morphology of the composite nanoparticles and the detection process were characterized by means of SEM, TEM, CV, EIS, DPV, fluorescence spectroscopy and laser confocal imaging. Under the optimized conditions, the proposed method exhibited a wide detection range of 0.05–300 μmol/L, with a detection limit of 0.01 μmol/L. Moreover, the modified electrode demonstrated satisfactory regenerability, and the proposed method showed excellent reproducibility and stability. The development platform could offer a new strategy for real-time assessment of drug concentration within cultured breast cancer cells in vitro. Full article

Background/Objectives: Eye infection with herpes simplex virus type 1 (HSV-1) can result in keratitis, a leading cause of corneal blindness. We evaluated whether an experimental vaccine containing HSV-2 immunogens to prevent genital herpes also protects against HSV-1 eye infection and neuroinvasion. Methods: Mice were immunized twice, one month apart, with PBS or a nucleoside-modified lipid nanoparticle vaccine containing mRNA encoding for gC2, gD2, and gE2. One month later, 10⁶ plaque forming units (PFU) (10 lethal dose 50, LD₅₀) of the HSV-1 McKrae strain were added to the intact cornea of each eye. Results: The vaccine prevented death and markedly reduced eyelid and attached conjunctival inflammation (blepharoconjunctivitis) and weight loss compared with the PBS group. Tissues from the ocular conjunctiva and eye bulb, olfactory bulb/peduncle, trigeminal ganglia, and brain (brainstem, cerebrum, and cerebellum) were harvested 5 days post-infection from 5 mice each in the PBS and vaccine groups, and from another 10 mice in the vaccine group 7 weeks post-infection. At 5 days, HSV-1 was not detected in any tissue in the vaccine group, while viral titers were positive in 16 of 25 (64%), and HSV-1 DNA was detected in 22 of 25 (88%) individual tissues in the PBS group. Histopathological and immunohistochemical analysis at 5 days post-infection confirmed that the vaccine protected against inflammation; however, some animals experienced breakthrough blepharoconjunctivitis. At 7 weeks, 3 of 10 (30%) mice in the vaccine group had HSV-1 DNA detected in the eyes or trigeminal ganglia tissues, but no animal had HSV-1 DNA detected in brain tissues. The vaccine produced cross-reactive HSV-1 neutralizing antibodies and gD1 IgG binding antibodies, but low or undetectable cross-reactive binding antibodies to gC1 and gE1. Conclusions: Despite occasional mild, localized breakthrough infections, the vaccine provided disease-modifying immunity and was neuroprotective. The results suggest that a single herpes vaccine effective against genital HSV-2 may be neuroprotective against HSV-1 following eye infection. Full article

Background: Silver nanoparticles (AgNPs) trigger apoptosis in cancer cells, while albumin nanoparticles enable effective drug delivery. This study compares the antitumor and cytotoxic effects of albumin-coated AgNPs (AgNPs-Alb) versus AgNPs on human prostate cancer cell lines. Method: AgNPs-Alb were synthesized and tested against PC3 and LNCaP prostate cancer cell lines. Characterization via Transmission Electron Microscopy (TEM), Dynamic Light Scattering (DLS), and Ultraviolet-Visible (UV-Vis) spectroscopy confirmed their properties. IC50 values were determined using MTT assay, with apoptosis assessed by Annexin-V/PI staining. DNA cell cycle was analyzed by PI staining. Migration, proliferation, and nuclear morphology were evaluated through scratch-wound, colony-forming, and Hoechst staining assays. Gene expression of Snail, E-cadherin, VEGF-C, VEGF-A, Bcl2, Bax, and P53 was analyzed using real-time PCR. Results: The IC50 values for AgNPs and AgNPs-Alb were 48 μM and 32 μM in PC3 cells, and 110 μM and 95 μM in LNCaP cells, respectively. AgNPs-Alb significantly inhibited PC3 cell migration compared to AgNPs (p < 0.001) and Bicalutamide (p < 0.0001). In both cell lines, AgNPs-Alb significantly reduced colony formation compared to AgNPs and Bicalutamide (p < 0.05). Flow cytometry revealed a higher percentage of apoptotic cells in PC3 with AgNPs-Alb treatment compared to AgNPs and Bicalutamide. In LNCaP cells, AgNPs-Alb induced a significantly higher percentage of Sub-G1 cells. AgNPs-Alb treatment caused greater mRNA suppression of VEGF-A and a higher Bax/Bcl2 ratio in PC3 and LNCaP cells (p < 0.05). Additionally, a significant increase in P53 and E-cadherin, alongside a decrease in VEGF-C expression in LnCAP cells, was observed (p < 0.05). Conclusions: This study suggests that AgNPs-Alb have stronger anticancer and cytotoxic effects compared to AgNPs alone against PCa cell lines and higher effects were observed on PC3 cells compared to LnCAP cells. Full article

Background/Objectives: Leishmaniasis is a disease affecting millions of people caused by parasites of the genus Leishmania. The GP63 protein of Leishmania infantum (LiGP63) is one of its major surface antigens and a main virulence factor, playing a role in the adhesion of extracellular promastigote stages to macrophages and in the survival of intracellular amastigotes. Methods: Here, DNA aptamers have been developed against LiGP63 through the systematic evolution of ligands by exponential enrichment. Results: Twenty individual aptamer sequences were characterized using confocal fluorescence microscopy and flow cytometry analysis, and 14 of them had targeting to more than 70% of L. infantum promastigotes with different subcellular localization patterns. Subsequent dot blot analyses narrowed down the selection to five candidates for further characterization through an aptamer-linked immobilized sorbent assay where it was possible to detect endogenous LiGP63 in L. infantum promastigote lysates. The five selected aptamers recognized the recombinant LiGP63 protein with binding affinities ranging from 0.3 to 2.1 µM. Promastigotes preincubated with LiGP63Apt-4, -27 and -28 exhibited a significantly reduced adhesion to and infection of RAW 264.7 macrophages. Moreover, when LiGP63Apt-4 and -28 were conjugated to liposomes, these two aptamers significantly enhanced the targeting to L. infantum promastigotes compared to plain liposomes. Conclusions: Given their improved stability and cost-effectiveness over antibodies, the aptamers evolved here represent promising candidates for new therapeutic and diagnostic approaches and for future nanoparticle-based drug delivery strategies in leishmaniasis. Full article

Inspired by natural scaffolds, artificial scaffolds have garnered significant attention in recent years. Compared with synthetic scaffolds such as organic and polymer scaffolds, biological scaffolds from the foundational biomolecules nucleic acids (DNA/RNA) and proteins demonstrate distinct advantages in the assembly of inorganic nanoparticles and proteins, as well as in drug delivery. These advantages stem from their exquisite spatial structures, genetically encoded programmability, and their favorable biocompatibility, which is attributed to natural building blocks and degradable backbones that minimize long-term cytotoxicity. The intrinsic properties and structural symmetry of biomacromolecules as building blocks often determine the properties of the corresponding assemblies, and thus greatly influence their functions. In this review, we classify bottom-up constructed biological scaffolds according to these two primary constituent classes (nucleic acids and proteins) to examine their framework structures and key features. We also discuss the relevant applications of artificial bioscaffolds. As an emerging class of nanomaterial with precise structures and genetic programmability, biological scaffolds hold significant promise for future development. Full article

In this work, we designed non-viral gene delivery vector systems based on three poly(2-ethyl-2-oxazoline)-ran-poly[2-(3-butenyl)-2-oxazoline] copolymers functionalized by primary, secondary, and tertiary amino groups. The impact of copolymer structure and composition was sought through the examination of basic physicochemical and biological parameters. The complexation ability of copolymers with plasmid DNA was studied by ethidium bromide quenching assay. The polyplex particles size and ζ-potential were determined by dynamic and electrophoretic light scattering. The release ability of copolymers was assessed by competitive displacement of DNA using dextran sulfate. The biological performance of amino-functionalized poly(2-ethyl-2-oxazoline)-ran-poly[2-(3-butenyl)-2-oxazoline] based gene delivery systems was evaluated, and their behavior under various environmental conditions, such as pH and ionic strength, was investigated. Cytotoxicity was assessed in two human lung-derived cell lines, and the ability of the copolymers to mediate plasmid DNA delivery and expression was examined. The resulting polyplex nanoparticles exhibited the ability to release DNA molecules and sensitivity to alterations in pH and ionic strength. All systems showed high biocompatibility and were able to mediate plasmid DNA delivery, resulting in detectable EGFP expression in vitro. The vector properties were found to be driven by a multifactorial interplay among hydrophobic character, thermoresponsive behavior, polymer mobility, charge accessibility, intracellular environmental responsiveness, secondary structure effects, etc. The copolymer bearing primary amino groups displayed a distinct balance between DNA binding and release, characterized by moderate complex stability and enhanced sensitivity to environmental changes. These findings provide mechanistic insight into how amino functionality and polymer structure influence the structure–property–behavior relationships of polyoxazoline-based non-viral gene delivery systems. Full article

The global expansion of genetically modified (GM) crop cultivation has increased the demand for analytical platforms that can provide rapid, reliable, and cost-effective detection of GM-derived ingredients to support traceability, regulatory compliance, and accurate labeling. Conventional molecular assays such as polymerase chain reaction (PCR) and isothermal amplification are highly sensitive and specific but depend on sophisticated instrumentation and trained personnel, limiting their applicability in field settings. Here, we present a label-free and amplification-free nanobiosensor based on citrate-capped gold nanoparticles (AuNPs) for the direct colorimetric detection of the Cry1Ac gene associated with the MON87701 soybean event, without the use of polymerase chain reaction (PCR) or any enzymatic nucleic acid amplification step. The assay relies on the localized surface plasmon resonance (LSPR) of AuNPs, which induces a red-to-purple color transition upon hybridization between complementary DNA strands. Critical reaction parameters, including NaCl concentration, AuNP size, and ionic strength, were optimized to enable selective and reproducible aggregation. Integration with a Support Vector Machine (SVM) algorithm enabled automated spectral classification and semi-quantitative discrimination of GM content levels. The optimized AuNP–SVM system achieved high sensitivity (limit of detection ≈ 2.5 ng μL⁻¹, depending on nanoparticle batch), strong specificity toward Cry1Ac-positive sequences, and reproducible classification accuracies exceeding 90%. By eliminating enzymatic amplification steps, the proposed platform significantly reduces assay time, operational complexity, and instrumentation requirements, making it suitable for rapid on-site GMO screening. Full article

Glioblastoma multiforme (GBM) remains the most aggressive and treatment-refractory form of primary brain tumor in adults, characterized by rapid proliferation, intratumoral heterogeneity and resistance to current therapies. Despite therapeutic advancements in surgical resection, radiotherapy and chemotherapy, clinical outcomes remain poor, underscoring the need for innovative molecular strategies. This review examines the therapeutic potential of CRISPR/Cas9 genome-editing technologies in GBM, highlighting their ability to model, dissect and potentially correct the genetic alterations that drive GBM tumorigenesis. Key molecular targets, such as EGFR, PTEN, TP53, NF1 and PIK3CA, are discussed within the context of GBM’s mutational and signaling landscape. We further outline emerging CRISPR applications in preclinical models, the current status of CRISPR-based clinical trials and the major barriers hindering translation, including off-target effects, immunogenicity and the challenge of delivering gene-editing systems across the blood–brain barrier. Particular emphasis is placed on delivery technologies, viral and non-viral vectors, including lipid nanoparticles, polymeric systems, inorganic nanocarriers and DNA nanostructures, which are rapidly evolving to improve precision, safety and CNS penetrance. Collectively, this review highlights CRISPR/Cas9 as a powerful tool whose integration with molecular neuro-oncology and precision medicine may ultimately shift GBM treatment toward more personalized and durable therapeutic interventions. Full article

Salmonella remains a leading cause of foodborne illness worldwide, with poultry products representing a major source of human exposure, underscoring the need for rapid and reliable detection methods. Although qPCR offers sensitive and timely pathogen detection, assay performance is highly dependent on sample preparation efficiency and nucleic acid purity, particularly in complex food matrices. In this study, we systematically optimized the sample preparation workflow of a SYBR Green based qPCR assay for enrichment-free detection of Salmonella enterica in poultry. Multiple lysis chemistries, incubation times, DNA extraction methods, centrifugation strategies, inoculum sources, and magnetic nanoparticle (MNP) assisted workflows were evaluated using phosphate-buffered saline and chicken rinsate matrices. Among the conditions tested, lysis with 20 µL Proteinase K and 400 µL PrepMan™ for 20 min produced the lowest and most consistent Cq values. Although Promega Wizard^® produced slightly lower mean Cq values than PrepMan™, statistical analysis showed no significant differences between extraction methods or centrifugation protocols, indicating comparable overall performance. Broth-derived inocula yielded earlier and more reproducible Cq values than colony-derived preparations. In contrast, inclusion of MNP processing resulted in higher Cq values in both matrices compared to the non-MNP workflow. Overall, these findings demonstrate that optimized lysis, extraction, and centrifugation workflows enhances the consistency and analytical reliability of direct qPCR detection of Salmonella in poultry matrices, supporting laboratory-based rapid detection applications. Full article