Search Results (1,072)

Background/Objectives: The rapid expansion of New Approach Methodologies (NAMs) is transforming oral biopharmaceutics by offering mechanistically rich, human-relevant tools that can reduce reliance on animal testing while improving translational confidence. Regulatory agencies, including the Food and Drug Administration (FDA) and the European Medicines Agency (EMA), are increasingly open to NAM-generated evidence, provided that methods are fit-for-purpose and scientifically justified. This review synthesizes current advances and evaluates how NAMs can be integrated across drug-development stages to enhance the prediction of oral absorption, formulation performance, and regulatory decision-making. Methods: A comprehensive literature review was conducted across classical and emerging methodologies, including in vitro permeability and solubility models, organoids, organ-on-a-chip (OoC) systems, machine learning frameworks, and mechanistic approaches such as the physiologically based pharmacokinetic (PBPK) and biopharmaceutics (PBBM) models. Emphasis was placed on physiological relevance, predictive performance, validation status, and regulatory applicability. Results: Classical tools remain essential for the Biopharmaceutics Classification System (BCS)-based biowaivers and risk-based assessments, yet they often lack physiological fidelity. NAMs provide enhanced representation of intestinal architecture, hydrodynamics, transporter activity, and metabolism. Organoids and microphysiological systems generate high-quality permeability and metabolic data, while computational NAMs enable scalable prediction of ADME properties and formulation behavior. When integrated into PBPK/PBBM models, these methods have great potential in predicting in vivo performance in humans. Evidence demonstrates that NAMs can refine, reduce, and, in specific contexts, replace animal studies without compromising scientific rigor. Conclusions: NAMs complement, rather than displace, classical biopharmaceutic tools, enabling a more mechanistic, human-centered, and ethically responsible framework for drug development. Their effective implementation will depend on continued validation, standardization, and regulatory harmonization as the field transitions toward fully NAM-supported biopharmaceutical assessment. Full article

This study presents the first bioarcheological analysis of the Mnogovalikovaia culture (Middle Bronze Age) from the North Pontic region, evaluating six human skeletons (six adult males) discovered in tumuli at Novosiolovca, Burlăceni, and Ordășei (Republic of Moldova). Dental analysis reveals moderate-to-advanced occlusal wear, chipping on posterior teeth, and anterior teeth modifications indicating both dietary practices and extramasticatory activities. The characteristics of the dental morphology, including non-metric dental traits (i.e., degree of cusp expression, presence of supernumerary cusps, bilateral asymmetry and the occlusal surface shape of maxillary and mandibular second molars (M2) highlight population-specific features influenced by genetic and environmental factors. Traces of ochre on skeletons suggest specific funerary practices. The study fills a significant gap in the understanding of Mnogovalikovaia communities, providing important data on their biological and cultural characteristics, lifestyle, and funerary practices. These findings also represent a basis for future research on this population, requiring larger samples and biomolecular analysis. Full article

The detection and retrieval of specific single cells within a cell population is useful for elucidating cellular function as well as early-stage cancer diagnosis by detecting circulating tumor cells. Microcapillaries are used to retrieve specific single cells from cell populations; however, quick single-cell retrieval that firmly adheres to the substrate without damaging the cell is difficult. In this study, we propose a single-cell selective retrieval method using a cone-shaped light-responsive gas-generating polymer (LGP) microscaffold array chip. An LGP microscaffold array chip with cone-shaped LGP microscaffolds was fabricated without any special equipment. When human cervical cancer cells were spread on the LGP microscaffold array chip, adhesion was achieved, and single cells were arranged on up to 73.3% of the cone-shaped LGP microscaffolds. When low-toxicity ultraviolet A light was irradiated from the back of the LGP microscaffold array chip, only a single cell adhering to the cone-shaped LGP microscaffold was released by the generated N₂ gas bubbles. More than 90% of the retrieved cells adhered, spread, and could be cultured for over 24 h. In conclusion, the proposed method is a simple and quick single-cell retrieval method that requires only a conventional inverted fluorescence microscope. Full article

Nanogrooves provide instructive cues to cells in culture. Several nanofabrication techniques have been developed to create biomimetic substrates, advancing our understanding of cell adhesion. Their integration into nervous system models highlights the critical role of the extracellular matrix (ECM) in developing functional tissue constructs for in vitro platforms such as Brain-on-Chip (BoC) and Nervous System-on-Chip (NoC). This study presents a nanofabrication approach that integrates photolithography and microtransfer molding (μTM) to pattern nanogrooves using photocurable polymer NOA81 onto microelectrode array (MEA) plates. The resulting nanogrooves exhibited a pattern periodicity of 976 nm and a ridge width of 232 nm, as confirmed by scanning electron microscopy and atomic force microscopy. We assessed the biocompatibility and functional impact of these modified substrates using human induced pluripotent stem cell (hiPSC)-derived neuronal cultures. Neurons cultured on nanogroove-modified MEAs exhibited aligned neural processes due to the anisotropic surface features and expressed vivid spiking behavior and higher burst frequency compared to randomly cultured neuronal networks. In conclusion, the proposed fabrication technique integrates nanogrooves with commercial MEAs using a combination of microtransfer molding and photolithography, resulting in modified culture substrates that enhance spike activity and network organization, aiding in the development of more in vivo-like neural models. Full article

Background/Objectives: Hepatocellular carcinoma (HCC) remains a major malignancy with high incidence and mortality, in part due to its diverse etiology and intratumoral heterogeneity, which contributes to drug resistance and frequent recurrence. SALL1 (Spalt-Like Transcription Factor 1), a zinc-finger transcription factor, was reported to function as a tumor suppressor in several cancers, including breast cancer and glioma, and accumulating evidence support its involvement in tumor biology. In this study, the role of SALL1 in HCC was examined. Methods: Public RNA and protein databases derived from human HCC were interrogated. Western blotting quantification of clinical HCC for SALL1 levels was carried out. Cell culture and xenograft studies were performed using genetically modified HCC tumor cells. Results: As revealed by pubic RNA and protein database analysis and further western blotting quantification of clinical samples of HCC, SALL1 is decreased in human HCC. The effect of reduced SALL1 expression on the tumorigenic properties and transcriptional regulation in HCC was then examined. Knockdown of SALL1 in the HCC cell lines Huh7 and Hep3B, enhanced cell proliferation in vitro and accelerated tumor growth in a xenograft mouse model, suggesting that lower SALL1 expression increases cell proliferation and tumorigenesis in HCC. RNA-seq and ChIP analyses further identified three novel candidate target genes (SLC6A14, GABRG1, and AKR1B10), suggesting that SALL1 may exert a tumor-suppressive effect, at least in part, through negative regulation of these genes. Conclusions: These findings establish SALL1 as a possible tumor suppressor and provide new insights into the biological significance of SALL1 downregulation in HCC. SALL1 could be a candidate prognostic marker and a potential therapeutic target. Full article

Translating preclinical findings into effective clinical cancer immunotherapies remains a major challenge, mainly because conventional in vitro and animal models often fail to capture the complexity, dynamics, and species-specific features of human immune responses. Organ-on-a-chip (OoC) technologies that combine engineered tissue architectures with precisely controlled microfluidic transport provide human-relevant microphysiological platforms for mechanistic studies of immune–tumor interactions and evaluation of therapeutic efficacy and immunotoxicity under defined microenvironmental conditions. However, immune responses involve time-dependent and interconnected processes, including immune cell trafficking, cytokine programs, metabolic shifts, and cytolysis, that are not adequately resolved by static or endpoint assays. Engineering immune-competent OoC systems therefore requires coordinated design of platform architectures, immune cell incorporation strategies, and integrated measurement workflows capable of capturing dynamic and state-dependent responses. In this review, we summarize engineering strategies for building immune-competent OoC platforms for cancer immunotherapy, focusing on platform architectures, immune cell incorporation methods, and fit-for-purpose assay workflows. Emphasis is placed on embedded sensing modalities (e.g., cytokine, oxygen, and impedance readouts) that provide valuable kinetic and state-variable data. Finally, we discuss key translational challenges, including reproducibility, standardization, and benchmarking, and outline near-term priorities to accelerate the adoption of immune-competent OoC systems in immunotherapy research and development. Full article

The widening gap between System-on-Chip (SoC) design complexity and verification productivity has rendered traditional script-based automation insufficient. While Large Language Models (LLMs) offer promise for code synthesis, they typically fail in hardware verification contexts due to a lack of architectural consistency and an inability to reason about temporal signal semantics. This paper proposes a Context-Aware Verification Loop (CAVL) methodology, an iterative framework that integrates semantic project indexing with simulation-based feedback to achieve verification closure. Unlike static generation, CAVL employs a dynamic refinement cycle where compiler diagnostics, simulation logs, and functional coverage metrics serve as feedback signals to guide the AI agent. We validate this framework on a dual-mode I²C Universal Verification Methodology (UVM) environment as a representative case study. The experimental results indicate, within this single-protocol context, the framework’s capacity to (1) resolve complex signal-level contention issues through logic refactoring, (2) achieve complete functional coverage via directed test synthesis, and (3) maintain cross-file architectural consistency with reduced human intervention. This work presents an initial quantitative baseline for AI-driven Electronic Design Automation (EDA), suggesting that context-aware feedback loops offer a pathway toward restructuring the verification engineer’s role from implementation to architectural intent specification. Full article

Ants play a central role in seed dispersal and predation, shaping plant recruitment, yet their foraging behavior is increasingly influenced by anthropogenic food subsidies. In human-modified landscapes, processed food waste may disrupt ant–seed interactions by diverting foragers or altering activity patterns, but the extent and mechanisms of these effects across habitats remain unclear. We conducted three field experiments in Panama to test how common food residues affect seed removal by ants in urban and forest environments. Using oat seeds as standardized diaspores, we (1) tested whether potato chips surrounding seed depots reduced removal, (2) evaluated the effects of adjacent chips or cookies on removal rates, ant activity, and species composition, and (3) manipulated the distance between chips and seeds (0, 30, 60 cm) to distinguish behavioral distraction from physical obstruction. Across experiments, seeds near food residues were removed significantly more slowly than controls, approximately half as fast in both habitats, despite differences in ant assemblages. Ant activity near seeds declined in the presence of food, particularly in the urban site. Suppression of seed removal occurred at close range but disappeared at 60 cm. These findings indicate that food waste disrupts ant-mediated seed removal through fine-scale behavioral shifts across contrasting habitats. Full article

Bone marrow(BM) is the primary site of hematopoiesis, supporting the self-renewal and differentiation of hematopoietic stem cells (HSCs). Its function depends on a highly complex microenvironment composed of stromal cells, vascular networks, extracellular matrix components, and dynamic biophysical signals. Traditional two-dimensional culture systems and animal models fail to adequately recapitulate the spatial architecture and dynamic regulatory processes of the human bone marrow niche, thereby limiting in-depth investigations into hematopoietic regulatory mechanisms, disease pathogenesis, and drug-induced bone marrow toxicity. In recent years, advances in microphysiological systems (MPS) have provided novel engineering approaches for the in vitro reconstruction of the bone marrow microenvironment. This review systematically summarizes current construction strategies for bone marrow MPS, including three-dimensional self-organized bone marrow organoids and microfluidic bone marrow-on-a-chip platforms. Particular attention is given to the roles of key cellular components, biomaterial scaffolds, vascularized architectures, and dynamic perfusion systems in biomimetic bone marrow engineering. In addition, we discuss strategies for constructing more complex models, such as vascular niches, vascularized bone tissue constructs, and bone metastasis models. Bone marrow MPS more faithfully recapitulate the hematopoietic microenvironment and provide a physiologically relevant in vitro platform for hematopoietic research, disease modeling, and drug evaluation, thereby supporting future advances in precision and regenerative medicine. Full article

Microphysiological systems (MPSs) have emerged as promising platforms for drug discovery and in vitro pharmacological testing. MPSs aid to reproduce physiologically relevant microenvironments, in which controlled perfusion can play important role. In this study, an on-chip AC electrothermal (ACET) pump was developed for pulsatile perfusion in microfluidic cell culture systems. The proposed pump generates fluid motion through the interaction between an applied electric field and temperature-dependent gradients in the electrical properties of the fluid. Pulsatile perfusion was produced by periodic application of an AC voltage to the electrode array, and the pulsation cycle could be controlled electrically. The maximum flow velocity increased with the applied AC voltage, demonstrating tunable flow generation by the ACET pump. To evaluate the applicability of the developed system to cell culture, human mesenchymal stem cells (hMSCs) were cultured under pulsatile perfusion conditions for five days. The results showed that osteogenic differentiation under pulsatile perfusion was higher than that under static culture conditions. These findings demonstrate the potential of the proposed on-chip ACET pump as a simple and effective platform for generating physiologically relevant pulsatile perfusion in microphysiological systems. Full article

In response to the pressing need for miniaturized MEMS microphones in wearable technology and mobile devices, and to surmount the technical limitations inherent in conventional piezoelectric microphones, which typically depend on enlarging chip dimensions or decreasing stiffness to attain low resonance frequencies, this study introduces a novel piezoelectric MEMS microphone (PMM) design predicated on a flexibility-enhanced rotating structure. The proposed design utilizes an aluminum scandium nitride (Al_0.8Sc_0.2N) piezoelectric thin film with 20% scandium doping and incorporates four equivalent sensing units formed by four curved cutting lines centrally located on the chip. This configuration employs a nested arrangement of four cantilever beams to substantially increase vibration compliance, thereby effectively lowering the natural frequency without altering the chip’s external size. Three-dimensional finite element simulations reveal that, relative to traditional triangular cantilever beam architectures, the flexibility-enhanced rotating structure reduces the natural frequency from 15.6 kHz to 13.49 kHz while enhancing sensitivity from −44.6 dB to −40 dB. The device was fabricated via a comprehensive microfabrication process and subsequently characterized within a standardized acoustic testing environment. Experimental results indicate that the microphone attains a sensitivity of −43.84 dB at 1 kHz and exhibits a first resonance frequency of 13.5 kHz, closely aligning with simulation predictions. Furthermore, the signal-to-noise ratio (SNR) reaches 58.3 dB across the full range of human-audible frequencies. By leveraging the flexibility-enhanced rotating structure, this work achieves an optimal compromise between elevated sensitivity and reduced resonance frequency within a compact form factor, thereby offering a viable technical solution for the advancement of high-performance miniature acoustic sensors. Full article

Metastatic breast cancer (MBC) remains an overwhelming clinical challenge due to its inherent clonal evolution and the frequent development of drug resistance. A significant hurdle in therapeutic discovery is the reliance on traditional 2D cell cultures and animal models, which often fail to accurately replicate human tumor pathophysiology or predict clinical responses. Consequently, the field of oncology is increasingly exploring a transition towards human-centric research that prioritizes biological data derived directly from patients. Considering the FDA Modernization Act 2.0 and the 2025 FDA Roadmap, frameworks are being established to explore the integration of new approach methodologies (NAMs)—including patient-derived organoids (PDOs) and organ-on-a-chip (OoC) systems—into the drug development pipeline. This review examines how these platforms aim to better simulate the human physiological environment by capturing the complex architecture and microenvironment of the tumor. We further discuss how the integration of these models with Artificial Intelligence (AI), spatial multi-omics, and real-time liquid biopsies is being investigated to enhance the speed and precision of therapeutic testing. While still in the translational phase, emerging evidence suggests that human-centric platforms may eventually support rapid functional drug screening, potentially informing patient treatment responses within clinically relevant timeframes. Strengthening the biological link between the patient and their longitudinal data represents a promising strategy to address the complexities of MBC and improve clinical outcomes. These human-centric platforms preserve patient-specific tumor heterogeneity, recapitulate microenvironmental interactions, and enable functional drug testing under physiologically relevant conditions, thereby improving translational accuracy compared to conventional models. Full article

Background/Objectives: Antibodies are a rapidly expanding field in drug discovery, but their monospecificity limits therapeutic applications, particularly in complex inflammatory diseases. Multispecific therapeutics, which combine variable regions targeting two or more antigens, offer potential advantages such as enhanced efficacy, broader target modulation, and reduced side effects. This study aimed to identify and characterize bispecific, VHH-based antibodies simultaneously targeting IL-6 and IL-17A—two key cytokines involved in autoimmune and chronic inflammatory conditions. Methods: A phage display screening was conducted using llama-derived VHH libraries to select binders against human IL-6 and IL-17A. Binding affinities of individual VHHs and assembled bispecific constructs were assessed using Bio-Layer Interferometry (BLI). Functional activity was evaluated using reporter cell lines responsive to IL-6 and IL-17A signaling. Biophysical and quality assessments of selected VHHs and bispecific antibodies were performed using the Uncle screening platform and LabChip capillary electrophoresis. Results: Several high-affinity VHH binders were identified for both IL-6 and IL-17A, and incorporated into bispecific antibody formats. The bispecific candidates exhibited simultaneous inhibition of both cytokine pathways in functional reporter assays. Biophysical characterization confirmed good stability and purity profiles for selected molecules. Conclusions: This study demonstrates the feasibility of generating stable, functional bispecific VHH-based antibodies targeting IL-6 and IL-17A. These constructs show potential as therapeutic agents for treating autoimmune and chronic inflammatory diseases by modulating multiple signaling pathways simultaneously. Full article

Background/Objectives: Epigenetic modifications, particularly DNA methylation, contribute to tumor progression and therapy resistance. Guadecitabine, a hypomethylating agent (HMA), has shown promising clinical activity when combined with carboplatin in preclinical models. We evaluated the combination of guadecitabine with carboplatin as a second-line treatment for extensive-stage small-cell lung cancer (SCLC; NCT03913455), one of the deadliest malignancies. Here, we report methylome changes in peripheral blood mononuclear cells (PBMCs) collected at baseline and during treatment from patients on the trial. Methods: PMBC DNA was analyzed using Infinium HumanMethylationEPIC v1.0 bead chips. Data were processed, and differentially methylated positions (DMPs) were identified and analyzed for pathway enrichment using bioinformatic approaches, and immune deconvolution analyses were conducted to investigate the impact on immune cell composition. Results: Direct comparison of PBMCs between cycle 2 day 5 (C2D5; post-treatment) vs. cycle 1 day 1 (C1D1; pre-treatment) revealed a greater number of hypomethylated DMPs (380 DMPs in C2D5 vs. C1D1 PBMCs; p < 0.05, |β| > 20%). Moreover, when first compared with normal PBMCs from cancer-free controls, the number of hypomethylated DMPs was even greater in C2D5 than in C1D1 (1771 vs. 237 DMPs, respectively; p < 0.05, |β| > 20%). Long interspersed nucleotide elements-1 (LINE-1) were significantly hypomethylated in PBMCs after HMA treatment (C2D5 vs. C1D1). Pathway analysis of hypomethylated DMPs revealed significant alterations in key signaling pathways, including NF-κB, Rho GTPase, and pulmonary fibrosis in C1D1 vs. C2D5. Normal PBMCs to C1D1 PBMCs revealed changes in IL-3 signaling, Fcγ receptor-mediated phagocytosis, and molecular mechanisms of cancer. Deconvolution analysis revealed a greater percentage of monocytes in C1D1 vs. normal PBMCs; after HMA treatment, percentages of monocytes and B cells decreased, while the eosinophil percentage increased in C1D1 vs. C2D5. Conclusions: HMA treatment has a global impact on PBMC methylomes in cancer patients. DNA methylation changes were associated with biological pathways related to PBMC function, and shifts in distinct immune cell populations were observed. Full article

One of the common issues in the R&D of new drugs is the failure of clinical trials caused by the species-specific inadequacy of animal models to assess drugs’ efficiency and safety. Therefore, systems like organ-on-a-chip and, particularly, liver-on-a-chip (LOC) can be an efficient tool for recapitulating in vivo-like human physiology at the microscale. This review focuses on discussing LOC design, emphasizing its architecture and validation to reveal the trends in searching for a balance between biomimetics and functionality. We found that the huge variety of already published models can be divided into five groups based on their configuration complexity: flat one-channel, flat two-channel, vertically stacked multilayered, hexagonal-patterned, and multi-well chips. While researchers attempt to recapitulate the liver’s histology and its functions in detail by increasing the complexity of devices’ architectonics, industrial companies prefer to promote more simple and flexible solutions. Thus, the LOC designs of the future require neglecting some liver characteristics to make them standardizable and sustainable, which could facilitate their introduction into the market and clinics. Full article