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Search Results (1,731)

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21 pages, 810 KB  
Article
Enhancing Quantum Literacy in Secondary Education Through Quantum Computing and Quantum Key Distribution
by Aspasia V. Oikonomou, Ilias K. Savvas and Omiros Iatrellis
Educ. Sci. 2025, 15(9), 1167; https://doi.org/10.3390/educsci15091167 (registering DOI) - 6 Sep 2025
Abstract
In the current era of rapid technological change, where artificial intelligence and quantum computing are reshaping knowledge, quantum literacy in high schools is becoming increasingly relevant. An understanding of quantum science is now important for fostering future readiness to prepare students for the [...] Read more.
In the current era of rapid technological change, where artificial intelligence and quantum computing are reshaping knowledge, quantum literacy in high schools is becoming increasingly relevant. An understanding of quantum science is now important for fostering future readiness to prepare students for the future, as it directly affects research, technology and innovation. Introducing quantum computing through educational tools and interactive platforms in schools will make quantum science accessible, equipping students with the necessary skills to understand and participate in future developments. This work investigates the necessity of quantum literacy among secondary education students, as well as their perceptions and understanding of basic concepts of quantum physics. Prior to data collection, students participated in two 90 min educational presentations that introduced fundamental principles of quantum physics through quantum computing and its applications, with an emphasis on cryptography and key distribution. Then, through the application of a specially designed questionnaire, data were collected from 78 students of different kind of schools and background and analyzed quantitatively and qualitatively. The results showed positive trends in students’ responses regarding their familiarity with quantum literacy and their understanding of fundamental principles such as superposition and entanglement. In addition, the analysis highlighted students’ interest in quantum computing and technology and its potential applications. This study highlights the need to integrate quantum literacy into the secondary education curriculum in order to foster scientific thinking and prepare students for the challenges of the quantum era. The educational intervention with the two presentations seemed to contribute positively to the development of students’ quantum literacy. Full article
30 pages, 6580 KB  
Article
Advanced Nanomaterial-Based Electrochemical Biosensing of Loop-Mediated Isothermal Amplification Products
by Ana Kuprešanin, Marija Pavlović, Ljiljana Šašić Zorić, Milinko Perić, Stefan Jarić, Teodora Knežić, Ljiljana Janjušević, Zorica Novaković, Marko Radović, Mila Djisalov, Nikola Kanas, Jovana Paskaš and Zoran Pavlović
Biosensors 2025, 15(9), 584; https://doi.org/10.3390/bios15090584 (registering DOI) - 5 Sep 2025
Abstract
The rapid and sensitive detection of regulatory elements within transgenic constructs of genetically modified organisms (GMOs) is essential for effective monitoring and control of their distribution. In this study, we present several innovative electrochemical biosensing platforms for the detection of regulatory sequences in [...] Read more.
The rapid and sensitive detection of regulatory elements within transgenic constructs of genetically modified organisms (GMOs) is essential for effective monitoring and control of their distribution. In this study, we present several innovative electrochemical biosensing platforms for the detection of regulatory sequences in genetically modified (GM) plants, combining the loop-mediated isothermal amplification (LAMP) method with electrodes functionalized by two-dimensional (2D) nanomaterials. The sensor design exploits the high surface area and excellent conductivity of reduced graphene oxide, Ti3C2Tx, and molybdenum disulfide (MoS2) to enhance signal transduction. Furthermore, we used a “green synthesis” method for Ti3C2Tx preparation that eliminates the use of hazardous hydrofluoric acid (HF) and hydrochloric acid (HCl), providing a safer and more sustainable approach for nanomaterial production. Within this framework, the performance of various custom-fabricated electrodes, including laser-patterned gold leaf films, physical vapor deposition (PVD)-deposited gold electrodes, and screen-printed gold electrodes, is evaluated and compared with commercial screen-printed gold electrodes. Additionally, gold and carbon electrodes were electrochemically covered by gold nanoparticles (AuNPs), and their properties were compared. Several electrochemical methods were used during the DNA detection, and their importance and differences in excitation signal were highlighted. Electrochemical properties, sensitivity, selectivity, and reproducibility are characterized for each electrode type to assess the influence of fabrication methods and material composition on sensor performance. The developed biosensing systems exhibit high sensitivity, specificity, and rapid response, highlighting their potential as practical tools for on-site GMO screening and regulatory compliance monitoring. This work advances electrochemical nucleic acid detection by integrating environmentally-friendly nanomaterial synthesis with robust biosensing technology. Full article
(This article belongs to the Section Biosensor Materials)
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23 pages, 5967 KB  
Article
Performance Evaluation of a HBsAg-Specific Immunoadsorbent Based on a Humanized Anti-HBsAg Monoclonal Antibody
by Shuangshuang Gao, Xiaobin Cai, Tianhui Yan, Yefu Wang and Xinyuan Tao
Biomedicines 2025, 13(9), 2175; https://doi.org/10.3390/biomedicines13092175 - 5 Sep 2025
Abstract
Background/Objectives: Hepatitis B virus (HBV) infection poses a major global health challenge, with current therapies like nucleos(t)ide analogs and pegylated interferon alpha offering limited functional cure rates due to persistent HBsAg-driven immune tolerance. This study aimed to develop a targeted immunoadsorption system [...] Read more.
Background/Objectives: Hepatitis B virus (HBV) infection poses a major global health challenge, with current therapies like nucleos(t)ide analogs and pegylated interferon alpha offering limited functional cure rates due to persistent HBsAg-driven immune tolerance. This study aimed to develop a targeted immunoadsorption system using a high-affinity humanized anti-HBsAg monoclonal antibody for efficient HBsAg and viral particle clearance, providing a novel approach to overcome therapeutic bottlenecks in chronic hepatitis B (CHB). Methods: A murine anti-HBsAg monoclonal antibody was humanized via complementarity-determining region grafting, resulting in HmAb-12 (equilibrium dissociation constant, KD = 0.36 nM). A stable Chinese Hamster Ovary K1 (CHO-K1) cell line was established for high-yield expression (fed-batch yield: 8.31 g/L). The antibody was covalently coupled to agarose microspheres (coupling efficiency > 95%) to prepare the immunoadsorbent. Efficacy was evaluated through in vitro dynamic circulation assays with artificial sera and preclinical trials using an integrated blood purification system in two CHB participants. Clearance rates for HBsAg and HBV DNA were quantified, with safety assessed via blood component monitoring. Results: In vitro, a single treatment cycle achieved HBsAg clearance rates of 70.14% (high antigen load, >105 IU/mL) and 92.10% (low antigen load, ~3000 IU/mL). Preclinically, one treatment session resulted in acute HBsAg reductions of 78.30% and 74.31% in participants with high and moderate antigen loads, respectively, alongside HBV DNA decreases of 65.66% and 73.55%. Minimal fluctuations in total protein and albumin levels (<15%) confirmed favorable safety profiles, with no serious adverse events observed. Conclusions: Preliminary findings from this study indicate that the HBsAg-specific immunoadsorption system can achieve efficient HBV antigen clearance with an initial favorable safety profile in a small cohort. These results support its further investigation as a potential therapeutic strategy for functional cure in CHB. Future work will focus on validating these findings in larger studies and exploring the system’s combinatory potential with existing blood purification platforms. Full article
(This article belongs to the Section Immunology and Immunotherapy)
21 pages, 5984 KB  
Article
Chrysin-Loaded Micelles Regulate Cell Cycle and Induce Intrinsic and Extrinsic Apoptosis in Ovarian Cancer Cells
by Serife Cakir, Ummugulsum Yildiz, Turgay Yildirim and Omer Aydin
Nanomaterials 2025, 15(17), 1362; https://doi.org/10.3390/nano15171362 - 4 Sep 2025
Viewed by 57
Abstract
Effective intracellular delivery for ovarian cancer therapy remains a significant challenge. We present chrysin-loaded p(MMA-co-DMAEMA)-b-(OEGMA-co-DMA), PMOD-Chr, a nanoparticle platform precisely engineered via RAFT polymerization for advanced therapeutic delivery. This multi-functional platform features a hydrophobic p(MMA) core encapsulating chrysin (Chr), a pH-responsive p(DMAEMA) segment [...] Read more.
Effective intracellular delivery for ovarian cancer therapy remains a significant challenge. We present chrysin-loaded p(MMA-co-DMAEMA)-b-(OEGMA-co-DMA), PMOD-Chr, a nanoparticle platform precisely engineered via RAFT polymerization for advanced therapeutic delivery. This multi-functional platform features a hydrophobic p(MMA) core encapsulating chrysin (Chr), a pH-responsive p(DMAEMA) segment for endosomal escape, and a hydrophilic OEGMA (Oligo(ethylene glycol) methyl ether methacrylate) shell functionalized for enhanced cellular affinity and systemic stability. The combination of OEGMA and DMA (Dopamine methacrylamide) block facilitates passive targeting of ovarian cancer cells, enhancing internalization. Nanoparticles prepared via the nanoprecipitation method exhibited ~220 nm, demonstrating effective size modulation along with high homogeneity and spherical morphology. In A2780 and OVCAR3 ovarian cancer cells, PMOD-Chr demonstrated significantly enhanced cytotoxicity, substantially lowering the effective IC50 dose of Chr. Mechanistically, PMOD-Chr induced a potent G2/M cell cycle arrest, driven by the upregulation of the CDK1/Cyclin B1 complex. Furthermore, the formulation potently triggered programmed cell death by concurrently activating both the intrinsic apoptotic pathway, evidenced by the modulation of Bax, Bcl2, and caspase 9, and the extrinsic pathway involving caspase 8. These findings emphasize that precision engineering via RAFT polymerization enables the creation of sophisticated, multi-stage nanomedicines that effectively overcome key delivery barriers, offering a highly promising targeted strategy for ovarian cancer. Full article
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29 pages, 1371 KB  
Review
Preparation of Multilayer Platforms for Advanced Wound Care Management
by Amir Mohammad Sharafi, Sina Pakkhesal, Farnaz Monajjemzadeh, Nastaran Alipour and Samin Hamidi
Polymers 2025, 17(17), 2393; https://doi.org/10.3390/polym17172393 - 2 Sep 2025
Viewed by 247
Abstract
Multilayer platforms have emerged as promising tools in the field of wound healing, offering a multifaceted approach to promote effective and accelerated tissue regeneration. This review article aims to provide a comprehensive overview of the various multilayer platforms employed in wound healing applications, [...] Read more.
Multilayer platforms have emerged as promising tools in the field of wound healing, offering a multifaceted approach to promote effective and accelerated tissue regeneration. This review article aims to provide a comprehensive overview of the various multilayer platforms employed in wound healing applications, highlighting their structure, fabrication methods, and potential mechanisms of action. The first section of the review focuses on the design and composition of multilayer platforms, encompassing different materials such as polymers, hydrogels, and biocompatible scaffolds. It discusses the significance of each layer in terms of its specific functionalities, including cell adhesion, drug/bioactive factor loading, antimicrobial properties, and mechanical support. The second section of the review delves into the mechanisms of action associated with multilayer platforms in wound healing. It discusses how these platforms facilitate wound closure, promote angiogenesis, modulate inflammation, and enhance tissue regeneration. The article also examines the role of multilayer platforms in providing a physical barrier against external pathogens, reducing the risk of infection, and creating a favorable microenvironment for wound healing. Overall, this review highlights the significant advancements made in the field of multilayer platforms for wound healing and underscores their potential as versatile therapeutic strategies. Full article
(This article belongs to the Section Biobased and Biodegradable Polymers)
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8 pages, 699 KB  
Communication
Ethylenediaminetetraacetic Acid (EDTA)-Decalcified, Formalin-Fixed Paraffin-Embedded (FFPE) Tumor Tissue Shows Comparable Quality and Quantity of DNA to Non-Decalcified Tissue in Next-Generation Sequencing (NGS)
by Francis Hong Xin Yap, Jen-Hwei Sng, Jeremy Wee Kiat Ng, Hanis Abdul Kadir, Pei Yi Chan and Timothy Kwang Yong Tay
J. Mol. Pathol. 2025, 6(3), 21; https://doi.org/10.3390/jmp6030021 - 2 Sep 2025
Viewed by 149
Abstract
Background: Bone tissue decalcification is essential for histopathological evaluation, but conventional methods using inorganic acids degrade nucleic acids, limiting molecular testing. EDTA is known to better preserve DNA, but its suitability for next-generation sequencing (NGS) in clinical settings remains to be validated. Methods: [...] Read more.
Background: Bone tissue decalcification is essential for histopathological evaluation, but conventional methods using inorganic acids degrade nucleic acids, limiting molecular testing. EDTA is known to better preserve DNA, but its suitability for next-generation sequencing (NGS) in clinical settings remains to be validated. Methods: This retrospective study evaluated 752 formalin-fixed paraffin-embedded (FFPE) tissue samples undergoing NGS between January 2022 and October 2024. Of these, 31 were decalcified using EDTA (Osteosoft, Merck, Germany). DNA was extracted using the Qiagen AllPrep® kit and quantified using Qubit and NanoDrop. Libraries were prepared with a custom 30-gene Ampliseq panel and sequenced on the Ion Torrent platform. Sequencing was deemed suboptimal if <95% of target regions reached ≥250X depth. Results were compared to 721 non-decalcified FFPE samples. Results: Suboptimal sequencing occurred in 9.7% of EDTA-decalcified and 9.0% of non-decalcified cases (p = 0.9). DNA concentration (Qubit) and NanoDrop 260/280 ratios were not significantly different (p = 0.4 and p = 0.8, respectively), though EDTA cases had lower DNA concentrations (NanoDrop, p = 0.006) and 260/230 ratios (p = 0.002). Mutation detection in decalcified samples was consistent with known mutation profiles for respective tumor types. Conclusions: EDTA-decalcified FFPE bone tissues produce NGS results comparable to non-decalcified specimens, with similar sequencing success rates and acceptable DNA quality. These findings support the use of EDTA as a suitable decalcification method for molecular diagnostics, enabling broader inclusion of bone specimens in clinical testing. Full article
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20 pages, 2264 KB  
Article
Development and Characterization of Citalopram-Loaded Thermosensitive Polymeric Micelles for Nasal Administration
by Fatima Rajab, Bence Sipos, Gábor Katona and Ildikó Csóka
Pharmaceutics 2025, 17(9), 1147; https://doi.org/10.3390/pharmaceutics17091147 - 1 Sep 2025
Viewed by 242
Abstract
Background/Objectives: The intranasal (IN) route of administration is a promising non-invasive approach for brain targeting, bypassing the blood–brain barrier and enhancing bioavailability. Citalopram hydrobromide (CT), a widely prescribed sparingly water-soluble selective serotonin reuptake inhibitor (SSRI), faces challenges with oral and intravenous administration, including [...] Read more.
Background/Objectives: The intranasal (IN) route of administration is a promising non-invasive approach for brain targeting, bypassing the blood–brain barrier and enhancing bioavailability. Citalopram hydrobromide (CT), a widely prescribed sparingly water-soluble selective serotonin reuptake inhibitor (SSRI), faces challenges with oral and intravenous administration, including delayed onset, adverse effects, and patient compliance issues. Methods: This study aimed to develop a novel thermoresponsive polymeric micelle (PM) system based on Pluronic® copolymers (Pluronic F127 and Poloxamer 188) improving CT’s solubility, stability, and nasal permeability for enhanced antidepressant efficacy. A preliminary study was conducted to select the optimized formulation. The preparation process involved using the thin-film hydration method, followed by freeze-drying. Comprehensive evaluations of optimized formulation characteristics included Z-average, polydispersity index (PdI), thermal behavior (lower critical solution temperature, LCST), encapsulation efficiency, X-ray powder diffraction (XRPD), thermodynamic solubility, and biological stability. Additionally, in vitro CT release and CT permeability in nasal conditions were studied. Stability under storage was also evaluated. Results: The optimized CT-PM formulation showed nanoscale micelle size (Z-average of 31.41 ± 0.99 nm), narrow size distribution (polydispersity index = 0.241), and a suitable thermal behavior for intranasal delivery (lower critical solution temperature (LCST) ~31 °C). Encapsulation efficiency reached approximately 90%, with an amorphous structure confirmed via XRPD, leading to a 95-fold increase in CT solubility. The formulation demonstrated appropriate biological and physical stability. In vitro studies showed a 25-fold faster CT release from optimized formulation compared to the initial CT, while CT-PM permeability in nasal conditions increased four-fold. Conclusions: This novel nanoscale thermosensitive formulation is a value-added strategy for nasal drug delivery systems, offering enhanced drug solubility, rapid drug release, stability, and improved permeability. This smart nanosystem represents a promising platform to overcome the limitations of conventional CT administration, improving therapeutic outcomes and patient compliance in depression management. Full article
(This article belongs to the Section Nanomedicine and Nanotechnology)
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16 pages, 519 KB  
Article
Pharmacy Students’ Experience of an Inaugural Lecture on Intercultural Competence
by Atta Abbas Naqvi, Merhawi Samsom, Lucy Watson and Hung Nguyen
Pharmacy 2025, 13(5), 122; https://doi.org/10.3390/pharmacy13050122 - 1 Sep 2025
Viewed by 227
Abstract
Background: Pharmacy schools in the United Kingdom (UK) are required by the regulator to train pharmacy students to be culturally competent. To meet this requirement, the Reading School of Pharmacy (RSoP) incorporated an inaugural, stand-alone, introductory session on intercultural competency. This study aimed [...] Read more.
Background: Pharmacy schools in the United Kingdom (UK) are required by the regulator to train pharmacy students to be culturally competent. To meet this requirement, the Reading School of Pharmacy (RSoP) incorporated an inaugural, stand-alone, introductory session on intercultural competency. This study aimed to gather students’ experiences of the lecture. Methods: A qualitative study documented the experiences of students in Years 2 and 3 of the Master of Pharmacy (MPharm) at the RSoP from 15 September to 31 December 2023. Semi-structured interviews were conducted online via Microsoft Teams®. A demographic form was prepared and sent as an online survey link on the Online Surveys® platform. All eligible students were invited to participate in the study via student mailing lists. An interview guide was prepared. Thematic analysis was conducted to identify key themes related to students’ awareness, the perceived importance of the subject in healthcare, and students’ preferred learning methods. The transcripts were coded, and similar codes were grouped to form sub-themes and themes. The study was approved by a research ethics committee. Results: A total of 11 students attended the interviews. Three major themes emerged: (1) awareness of and reflection on cultural competence, (2) understanding cultural competence and its importance, and (3) student-preferred pedagogy. The students suggested incorporating workshops and simulation-based assessments. Conclusions: MPharm pharmacy students at the RSoP appear to be receptive to new educational interventions aimed at enhancing cultural competence. They prefer practice-based learning and assessment methods when it comes to developing this skill. Full article
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5 pages, 585 KB  
Short Note
3-([4-(Acetylamino)phenyl]methoxy-1-carbonyl)-7-oxabicyclo[2.2.1]heptane-2-carboxylic Acid
by Kathryn N. Mayeaux, Bailey N. Baxter, Hannah K. Lawley, Caleb N. Lopansri, Mary Helene Marmande, Lucy A. Orr and David C. Forbes
Molbank 2025, 2025(3), M2056; https://doi.org/10.3390/M2056 - 30 Aug 2025
Viewed by 245
Abstract
Overexpression of protein phosphatase 5 (PP5) is linked to tumor cell growth, making it a candidate for small-molecule drug therapy. Since the PP2A domain has been selectively inhibited using functionalized scaffolds that maximize contacts, a similar approach is proposed to work for PP5. [...] Read more.
Overexpression of protein phosphatase 5 (PP5) is linked to tumor cell growth, making it a candidate for small-molecule drug therapy. Since the PP2A domain has been selectively inhibited using functionalized scaffolds that maximize contacts, a similar approach is proposed to work for PP5. As cantharidin’s demethylated cousin, norcantharidin, is a potent but unselective phosphatase inhibitor that can be prepared in just two synthetic steps, the bicyclic scaffold holds promise as an attractive target upon functionalization. Our hypothesis targets PP5 selectivity through derivatives of norcantharidin with functionalized attachments for optimal active-site binding. The methodology offers a promising platform for developing PP5-selective anticancer therapeutics. The approach reported herein exploits anhydride reactivity to yield a carboxylic acid derivative as our next-generation inhibitor of PP5. The methodology offers groundwork for future optimization of norcantharidin-based drug candidates with improved tumor selectivity, potency, and synthetic feasibility. Full article
(This article belongs to the Section Organic Synthesis and Biosynthesis)
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18 pages, 5211 KB  
Article
Data-Driven Performance-Oriented Rapid Process Optimization for 316 Stainless Steels Prepared by Laser Powder Bed Fusion
by Junyan Zhu, Meiling Jiang, Guoliang Huang and Ke Huang
Metals 2025, 15(9), 968; https://doi.org/10.3390/met15090968 - 30 Aug 2025
Viewed by 269
Abstract
In this work, a rapid process optimization framework for laser powder bed fusion (LPBF) based on a high-throughput mechanical testing platform and data analytical methods was proposed and validated. This framework enables the efficient building of a process–properties database and analytical model, as [...] Read more.
In this work, a rapid process optimization framework for laser powder bed fusion (LPBF) based on a high-throughput mechanical testing platform and data analytical methods was proposed and validated. This framework enables the efficient building of a process–properties database and analytical model, as well as the fine-tuning of customized mechanical properties. Unlike previous approaches that focused primarily on density as the main optimization target, this method directly aligns the mechanical properties by systematically varying the LPBF process parameters (e.g., laser power, scanning speed, etc.). Tensile specimens in the high densification range were prepared and tested using a high-throughput mechanical property test platform (HTP). Following this, an analytical model correlating tensile properties and process parameters was developed using response surface methodology (RSM). Based on this model recommendation, a specimen with a densification of 99.46% and a yield strength (YS) of 524.74 MPa was achieved, with only a 3.72% variation compared to the predicted value (526.08 MPa), confirming the model’s reliability. A comprehensive analysis of relative density, phase content and microstructure was conducted, comparing them with a specimen exhibiting lower properties. This study provides an effective method for the rapid evaluation and optimization of LPBF processing parameters for fine-tuning customized mechanical properties. Full article
(This article belongs to the Special Issue Welding and Additive Manufacturing of Metals)
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31 pages, 4538 KB  
Article
Ex Vivo Traceability Platform for Phospholipoproteomic Formulations: Functional Evidence Without Clinical Exposure
by Ramón Gutiérrez-Sandoval, Francisco Gutiérrez-Castro, Natalia Muñoz-Godoy, Ider Rivadeneira, Andy Lagos, Ignacio Muñoz, Jordan Iturra, Francisco Krakowiak, Cristián Peña-Vargas, Matías Vidal and Andrés Toledo
Biomedicines 2025, 13(9), 2101; https://doi.org/10.3390/biomedicines13092101 - 28 Aug 2025
Viewed by 271
Abstract
Background: Structurally active phospholipoproteomic formulations that lack pharmacodynamic targets or systemic absorption present unique challenges for validation. Designed for immune compatibility or structural modulation—rather than therapeutic effect—these platforms cannot be evaluated through conventional clinical or molecular frameworks. Methods: This study introduces a standardized, [...] Read more.
Background: Structurally active phospholipoproteomic formulations that lack pharmacodynamic targets or systemic absorption present unique challenges for validation. Designed for immune compatibility or structural modulation—rather than therapeutic effect—these platforms cannot be evaluated through conventional clinical or molecular frameworks. Methods: This study introduces a standardized, non-invasive ex vivo protocol using real-time kinetic imaging to document biological behavior under neutral conditions. Eight human tumor-derived adherent cell lines were selected for phenotypic stability and imaging compatibility. Phospholipoproteomic preparations were applied under harmonized conditions, and cellular responses were recorded continuously over 48 h. Results: Key parameters included signal continuity, morphological integrity, and inter-batch reproducibility. The system achieved high technical consistency without labeling, endpoint disruption, or destructive assays. Outputs included full kinetic curves and viability signals across multiple cell–fraction pairings. Conclusions: This method provides a regulatorily compatible foundation for functional documentation in non-pharmacodynamic programs where clinical trials are infeasible. It supports early-stage screening, batch comparability, and audit-ready records within SAP, CTD, or real-world evidence (RWE) ecosystems. By decoupling validation from systemic exposure, the protocol enables scalable, technically grounded decision-making for structurally defined immunobiological platforms. Full article
(This article belongs to the Special Issue New Trends in Cancer Immunotherapy)
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18 pages, 39111 KB  
Article
Impact of Beam Shape and Frequency on Weld Seam Geometry and Penetration Depth Using a Coherent Beam Combining Laser
by Karthik Ravi Krishna Murthy, Reza Sanei, Abhay Sharma, Simon Olschok and Uwe Reisgen
Appl. Sci. 2025, 15(17), 9432; https://doi.org/10.3390/app15179432 - 28 Aug 2025
Viewed by 317
Abstract
The geometry and quality of a weld seam are critical factors in laser beam welding, influencing mechanical performance and structural integrity. Dynamically modulated laser beams provide a precise means of tailoring energy input in high-power laser welding processes. This study investigates the influence [...] Read more.
The geometry and quality of a weld seam are critical factors in laser beam welding, influencing mechanical performance and structural integrity. Dynamically modulated laser beams provide a precise means of tailoring energy input in high-power laser welding processes. This study investigates the influence of beam shape and modulated frequency on weld seam geometry, penetration depth, and capillary behaviour using a coherent beam combining (CBC) laser system from Civan Lasers. Three beam intensity distributions—single point, line–point–line (LPL), and boomerang—were applied across a modulation frequency range of 1, 10, and 100 kHz during the welding of duplex and austenitic stainless steels. High-speed imaging captured real-time capillary dynamics, and the data were analysed to assess capillary stability, measure capillary diameter, and determine the capillary front angle as a function of frequency and beam shape. Transverse cross-sections of the welds were prepared to evaluate seam geometry and microstructure. The results show that beam shape significantly affects energy distribution and weld profile, while modulation frequency critically influences capillary behaviour and penetration characteristics. These findings highlight the critical role of dynamic beam shaping and frequency modulation in optimizing laser welding processes for material-specific performance, offering a versatile platform for advancing precision manufacturing using CBC technology. Full article
(This article belongs to the Special Issue Advanced Welding Technology and Its Applications)
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14 pages, 3301 KB  
Article
Optimized and Reliable Protoplast Isolation for Transient Gene Expression Studies in the Gymnosperm Tree Species Pinus densiflora
by Tram Thi Thu Nguyen, Na-Young Choi, Seung-Won Pyo, Young-Im Choi and Jae-Heung Ko
Forests 2025, 16(9), 1373; https://doi.org/10.3390/f16091373 - 26 Aug 2025
Viewed by 322
Abstract
Efficient protoplast isolation and gene transfection remain significant challenges in gymnosperms, particularly in Pinus species, where stable transformation is highly limited. Conventional pine protoplast preparation methods have resulted in extremely low transfection efficiencies, hindering functional genomic studies. This study presents an optimized method [...] Read more.
Efficient protoplast isolation and gene transfection remain significant challenges in gymnosperms, particularly in Pinus species, where stable transformation is highly limited. Conventional pine protoplast preparation methods have resulted in extremely low transfection efficiencies, hindering functional genomic studies. This study presents an optimized method for isolating high-yield, viable protoplasts from Pinus densiflora (Korean red pine), providing a robust system for transient gene expression assays. Splitting one-month-old cotyledons produced the highest mesophyll protoplast yield (5.0 × 106 cells/g FW), which further increased to 1.2 × 107 cells/g FW after optimizing the enzyme mixture (4.5% cellulase, 0.7% pectinase, 3% hemicellulase), maintaining viability above 86%. Developing xylem and whole-stem protoplasts were also successfully isolated by mitigating resin leakage and debris contamination, with a 17% sucrose gradient yielding 7.4 × 104 cells/g FW at 81.9% viability. Overcoming prior inefficiencies, this protocol significantly enhances gene transfection efficiency, achieving 94.1% GFP transformation with 82.9% viability. Furthermore, transient activation assays confirmed strong activation of pine-derived reporters by native effectors, underscoring the assay’s suitability for studying gymnosperm-specific gene regulation. Given the limited stable transformation strategies available for Pinus species, this optimized protoplast transient gene expression system provides a practical and reliable platform for transient gene expression analysis, offering valuable opportunities for studying gene function and regulation in gymnosperms. Full article
(This article belongs to the Section Genetics and Molecular Biology)
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20 pages, 72348 KB  
Article
Integrated Fluidic Platform for Washing and Mechanical Processing of Lipoaspirate for Downstream Fat Grafting and Regenerative Applications
by David Zalazar, Jiayi Feng, Derek A. Banyard, Marzieh Aliaghaei, Alan D. Widgerow and Jered B. Haun
Bioengineering 2025, 12(9), 918; https://doi.org/10.3390/bioengineering12090918 - 26 Aug 2025
Viewed by 579
Abstract
Autologous fat grafting of human lipoaspirate (LA) is increasingly used in reconstructive and cosmetic surgery for lipofilling and stem cell-rich “nanofat” reinjection for regenerative medicine. While commercial devices (e.g., REVOLVE and Puregraft) are available, many surgeons use non-standardized manual washing techniques, leading to [...] Read more.
Autologous fat grafting of human lipoaspirate (LA) is increasingly used in reconstructive and cosmetic surgery for lipofilling and stem cell-rich “nanofat” reinjection for regenerative medicine. While commercial devices (e.g., REVOLVE and Puregraft) are available, many surgeons use non-standardized manual washing techniques, leading to inconsistent graft retention (20–80%). Moreover, no system can unite washing directly with mechanical processing to produce a nanofat-like product directly from raw LA. We developed a novel preparation device (PD) that is designed for peristaltic pump-driven washing of LA and can be seamlessly combined with our previously developed Emulsification and Micronization Device (EMD) into an automated closed-loop platform. Human LA samples were washed with the PD and compared to standard manual washing via visual colorimetric analysis. We then evaluated the mechanical processing of PD-washed LA using our EMD and assessed cell count, viability, and stromal vascular fraction-derived subpopulations (i.e., mesenchymal stem cells, endothelial progenitor cells (EPCs), pericytes, transit-amplifying (TA) progenitor cells, and supra-adventitial adipose stromal cells). Recirculating LA through the PD for at least one minute resulted in sufficient mixing, producing LA with equivalent color and quality to manual washing. Integrating the EMD within a platform enabled both washing and mechanical processing under peristaltic flow, enriching key subpopulations compared to manual methods. Thus, our fluidic platform effectively washes LA in a closed-loop system, minimizing LA tissue manipulation and opportunity for contamination while also simplifying the workflow for mechanical processing. Further refinement and automation of this platform would enhance the reproducibility and quality of small-volume fat grafts, cell-assisted lipotransfer, and stem/progenitor cell injections to promote wound healing and angiogenesis. Full article
(This article belongs to the Special Issue Regenerative Technologies in Plastic and Reconstructive Surgery)
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12 pages, 1631 KB  
Article
Development of a Method for Producing Recombinant Human Granulocyte-Macrophage Colony-Stimulating Factor Using Fusion Protein Technology
by Ekaterina A. Volosnikova, Tatiana I. Esina, Natalia V. Volkova, Svetlana V. Belenkaya, Yana S. Gogina, Galina G. Shimina, Elena A. Vyazovaya, Svetlana G. Gamaley, Elena D. Danilenko and Dmitriy N. Shcherbakov
Curr. Issues Mol. Biol. 2025, 47(9), 681; https://doi.org/10.3390/cimb47090681 - 25 Aug 2025
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Abstract
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine with therapeutic applications in oncology and neurodegenerative diseases. However, its clinical use is limited by the high cost of eukaryotic production systems. Here, we developed a cost-effective Escherichia coli-based platform for high-yield production of [...] Read more.
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine with therapeutic applications in oncology and neurodegenerative diseases. However, its clinical use is limited by the high cost of eukaryotic production systems. Here, we developed a cost-effective Escherichia coli-based platform for high-yield production of biologically active recombinant human GM-CSF (rhGM-CSF) using SUMO fusion technology. The engineered pET-SUMO-GM plasmid enabled expression of a 33 kDa fusion protein, accounting for 23–25% of total cellular protein, though it primarily accumulated in inclusion bodies. A multi-step purification strategy—including nickel affinity chromatography, Ulp protease cleavage, and hydrophobic chromatography—yielded >99.5% pure rhGM-CSF. In vitro functional assays demonstrated equivalent activity to the WHO international standard (ED50: 0.045 vs. 0.043 ng/mL in TF-1 cell proliferation). In vivo, the preparation significantly restored neutrophil counts (3.4-fold increase, p ≤ 0.05) in a murine cyclophosphamide-induced myelosuppression model. Our results establish a scalable, prokaryotic-based method to produce functional rhGM-CSF, overcoming solubility and folding challenges while maintaining therapeutic efficacy. This approach could facilitate broader clinical and research applications of GM-CSF, particularly in resource-limited settings. Full article
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