Cell Behavior Research Articles

Alendronate sodium induces G1 phase arrest and apoptosis in human umbilical vein endothelial cells by inhibiting ROS-mediated ERK1/2 signaling

Bisphosphonates are potent bone resorption inhibitors, among which alendronate sodium (ALN) is commonly prescribed for most osteoporosis patients, but long-term application of ALN can cause bisphosphonate-related osteonecrosis of jaw (BRONJ), the pathogenesis of which remains unclear. Previous studies have suggested that bisphosphonates cause jaw ischemia by affecting the biological behavior of vascular endothelial cells, leading to BRONJ. However, the impacts of ALN on vascular endothelial cells and its mechanism remain unclear. The purpose of this work is to assess the influence of ALN on human umbilical vein endothelial cells (HUVECs) and clarify the molecular pathways involved. We found that high concentration of ALN induced G1 phase arrest in HUVECs, demonstrated by downregulation of Cyclin D1 and Cyclin D3. Moreover, high concentration of ALN treatment showed pro-apoptotic effect on HUVECs, demonstrated by increased levels of the cleaved caspase-3, the cleaved PARP and Bax, along with decreased levels of anti-apoptotic protein Bcl-2. Further experiments showed that ERK1/2 phosphorylation was decreased. Additionally, ALN provoked the build-up of reactive oxygen species (ROS) in HUVECs, leading to ERK1/2 pathway suppression. N-acetyl-L-cysteine (NAC), a ROS scavenger, efficiently promoted the ERK1/2 phosphorylation and mitigated the G1 phase arrest and apoptosis triggered by ALN in HUVECs. PD0325901, an inhibitor of ERK1/2 that diminishes the ERK1/2 phosphorylation enhanced the ALN-induced G1 phase arrest and apoptosis in HUVECs. These findings show that ALN induces G1 phase arrest and apoptosis through ROS-mediated ERK1/2 pathway inhibition in HUVECs, providing novel insights into the pathogenic process, prevention and treatment of BRONJ in individuals receiving extended use of ALN.

HTF4 modulates the transcription of GID2 to promote the malignant biological behavior of pancreatic cancer

BackgroundHelix-loop-helix transcription factor 4 (HTF4) as an anti-cancer target has been reported in many human cancers, but limited data exists regarding the effect of HTF4 in pancreatic cancer. In this study, we aimed to investigate the role of HTF4 in pancreatic cancer. MethodsThe expression levels of HTF4 in clinical pancreatic cancer samples were measured. HTF4 was knocked down or overexpressed in pancreatic cancer cells and was subsequently tested for bio-function using in vitro assays and in vivo. The regulation of HTF4 on GID2 was assessed via bioinformatic tools and dual-luciferase reporter assay. ResultsWe found that HTF4 was highly expressed in pancreatic cancer tissues and correlated with poor patient prognosis. In addition, knocking down HTF4 expression inhibited cell proliferation, migration, and invasion, whereas HTF4 overexpression exerted the opposite effect. Moreover, HTF4 promoted tumor growth and metastasis in pancreatic cancer. Further, HTF4 bound to the GID2 promoter region and promoted transcriptional activation of GID2 in pancreatic cancer cells. GID2 knockdown suppressed HTF4-induced malignant behaviors of pancreatic cancer cells. ConclusionsOur findings suggest that the HTF4/GID2 axis accelerates the progression of pancreatic cancer, providing a potential therapeutic target and prognostic indicator for the treatment of pancreatic cancer patients.

The effect of surface roughness and wettability on the adhesion and proliferation of Saos-2 cells seeded on 3D printed poly(3-hydroxybutyrate)/polylactide (PHB/PLA) surfaces

The aim of this work is to explore the effect of surface characteristics of the 3D printed scaffolds on their suitability for Saos-2 cells growth. Two promising materials for bone tissue engineering were examined, both containing poly(3-hydroxybutyrate)/poly(d,l-lactide) plasticized blend and one of them filled with bioactive tricalcium phosphate. The surface free energy (SFE) of samples was determined by contact angle measurement with four liquids. For non-filled sample the water contact angle (WCA) was 74° and its SFE was 40 mN m−1. The composite sample exhibited substantially decreased WCA, 61°. Moreover, the addition of the tricalcium phosphate caused doubling of the polar component of the SFE which increased by a total of 13%. 3D printed surfaces prepared by fused deposition modeling method showed a profound increase of WCA due to the increase in their surface roughness which was analyzed by confocal microscopy. The change in wetting properties strongly affected the behavior of Saos-2 cells on printed surfaces. The number of cells as measured by DNA quantification linearly decreased with increasing surface WCA. The highest number of cells was observed on the pressed samples and 3D printed surfaces made of simple lines and a grid with 50 μm gap between lines. The favored surfaces exhibit WCA under 80° which is important information for the future design of the scaffolds.

Experimental investigation of thermal runaway behavior and propagation inhibition of lithium-ion battery by immersion cooling

Understanding the impact of immersion cooling on thermal runaway (TR) behavior and its inhibitory effect on thermal runaway propagation (TRP) is crucial for its practical application. This study explores immersion cooling for individual cells and battery modules, examines the TR behavior of 18,650 single cells at various immersion depths and states of charge (SOC) using filament heating, and investigates TRP between cells. Immersion cooling with a depth of 70 mm significantly extends the interval between the rupture of the battery safety valve and the TR trigger from 70 s (at a 20 mm depth) to 312 s, while reducing the peak TR temperature to 306 °C. Additionally, cells with 100 %, 75 %, and 50 % SOC show that higher battery capacity increases the risk of TR and thermal hazards. TRP experiments demonstrate that immersion cooling can inhibit TRP between cells, though safety valves of adjacent cells in 100 %, 75 %, and 50 % SOC battery modules still ruptured, with peak temperatures reaching 118.2 °C, 121.4 °C, and 111.5 °C respectively, posing a potential TR risk. This study highlights the TR behavior of single cells at different immersion depths and confirms that immersion cooling can inhibit TRP, providing valuable insights for the future application of immersion cooling in lithium battery thermal management systems.

SULF1 Activates the VEGFR2/PI3K/AKT Pathway to Promote the Development of Cervical Cancer.

Sulfatase 1 (SULF1) can regulate the binding of numerous signaling molecules by removing 6-O-sulfate from heparan sulfate proteoglycans (HSPGs) to affect numerous physiological and pathological processes. Our research aimed to investigate the effect of the SULF1-mediated VEGFR2/PI3K/AKT signaling pathway on tumorigenesis and development of cervical cancer (CC). The expression and prognostic values of SULF1 in patients with CC were analyzed through bioinformatics analysis, qRT-PCR, immunohistochemistry, and western blot. The function and regulatory mechanism of SULF1 in proliferation, migration, and invasion of cervical cancer cells were examined through lentivirus transduction, CCK8, flow cytometry analysis, plate colony formation assay, scratch assay, transwell assay, western blot, VEGFR2 inhibitor (Ki8751), and mouse models. SULF1 expression was significantly upregulated in CC tissues, which was significantly associated with poor prognosis of patients with CC. In vitro, the upregulation of SULF1 expression in HeLa cells promoted cell proliferation, colony formation, migration, and invasion while inhibiting apoptosis. Conversely, the downregulation of SULF1 expression had the opposite effect. In vivo, the upregulation of SULF1 expression resulted in a significant increase in both tumor growth and angiogenesis, while its downregulation had the opposite effect. Furthermore, western blot detection and cell function rescue assay confirmed that the upregulation of SULF1 in HeLa cells promoted the tumorigenic behaviors of cancer cells by activating the VEGFR2/PI3K/AKT signaling pathway. SULF1 plays an oncogenic role in the tumorigenesis and development of CC, indicating its potential as a novel molecular target for gene-targeted therapy in patients with CC.

Matrix stiffness increases energy efficiency of endothelial cells

To form blood vessels, endothelial cells rearrange their cytoskeleton, generate traction stresses, migrate, and proliferate, all of which require energy. Despite these energetic costs, stiffening of the extracellular matrix promotes tumor angiogenesis and increases cell contractility. However, the interplay between extracellular matrix, cell contractility, and cellular energetics remains mechanistically unclear. Here, we utilized polyacrylamide substrates with various stiffnesses, a real-time biosensor of ATP, and traction force microscopy to show that endothelial cells exhibit increasing traction forces and energy usage trend as substrate stiffness increases. Inhibition of cytoskeleton reorganization via ROCK inhibition resulted in decreased cellular energy efficiency, and an opposite trend was found when cells were treated with manganese to promote integrin affinity. Altogether, our data reveal a link between matrix stiffness, cell contractility, and cell energetics, suggesting that endothelial cells on stiffer substrates can better convert intracellular energy into cellular traction forces. Given the critical role of cellular metabolism in cell function, our study also suggests that not only energy production but also the efficiency of its use plays a vital role in regulating cell behaviors and may help explain how increased matrix stiffness promotes angiogenesis.

Superposition of Substrate Deformation Fields Induced by Molecular Clutches Explains Cell Spatial Sensing of Ligands.

Cells can sense the physical properties of the extracellular matrices (ECMs), such as stiffness and ligand density, through cell adhesions to actively regulate their behaviors. Recent studies have shown that varying ligand spacing of ECMs can influence adhesion size, cell spreading, and even stem cell differentiation, indicating that cells have the spatial sensing ability of ECM ligands. However, the mechanism of the cells' spatial sensing remains unclear. In this study, we have developed a lattice-spring motor-clutch model by integrating cell membrane deformation, the talin unfolding mechanism, and the lattice spring for substrate ligand distribution to explore how the spatial distribution of integrin ligands and substrate stiffness influence cell spreading and adhesion dynamics. By applying the Gillespie algorithm, we found that large ligand spacing reduces the superposition effect of the substrate's displacement fields generated by pulling force from motor-clutch units, increasing the effective stiffness probed by the force-sensitive receptors; this finding explains a series of previous experiments. Furthermore, using the mean-field theory, we obtain the effective stiffness sensed by bound clutches analytically; our analysis shows that the bound clutch number and ligand spacing are the two key factors that affect the superposition effects of deformation fields and, hence, the effective stiffness. Overall, our study reveals the mechanism of cells' spatial sensing, i.e., ligand spacing changes the effective stiffness sensed by cells due to the superposition effect of deformation fields, which provides a physical clue for designing and developing biological materials that effectively control cell behavior and function.

Novel strategies in HPV‑16‑related cervical cancer treatment: An in vitro study of combined siRNA-E5 with oxaliplatin and ifosfamide chemotherapy.

Cervical cancer, primarily caused by HPV infection, remains a global health concern. Current treatments face challenges including drug resistance and toxicity. This study investigates combining E5-siRNA with chemotherapy drugs, Oxaliplatin and Ifosfamide, to enhance treatment efficacy in HPV-16 positive cervical cancer cells, targeting E5 oncoprotein to overcome limitations of existing therapies. The CaSki cervical cancer cell line was transfected with E5-siRNA, and subsequently treated with Oxaliplatin/Ifosfamide. Quantitative real-time PCR was employed to assess the expression of related genes including p53, MMP2, Nanog, and Caspases. Cell apoptosis, cell cycle progression, and cell viability were evaluated using Annexin V/PI staining, DAPI staining, and MTT test, respectively. Furthermore, stemness ability was determined through a colony formation assay, and cell motility was assessed by wound healing assay. E5-siRNA transfection significantly reduced E5 mRNA expression in CaSki cells compared to the control group. The MTT assay revealed that monotherapy with E5-siRNA, Oxaliplatin, or Ifosfamide had moderate effects on cell viability. However, combination therapy showed synergistic effects, reducing the IC50 of Oxaliplatin from 11.42 × 10-8 M (45.36 μg/ml) to 6.71 × 10-8 M (26.66 μg/ml) and Ifosfamide from 12.52 × 10-5 M (32.7 μg/ml) to 8.206 × 10-5 M (21.43 μg/ml). Flow cytometry analysis demonstrated a significant increase in apoptosis for combination treatments, with apoptosis rates rising from 11.02 % (Oxaliplatin alone) and 16.98 % (Ifosfamide alone) to 24.8 % (Oxaliplatin + E5-siRNA) and 34.9 % (Ifosfamide + E5-siRNA). The sub-G1 cell population increased from 15.7 % (Oxaliplatin alone) and 18 % (Ifosfamide alone) to 21.9 % (Oxaliplatin + E5-siRNA) and 27.1 % (Ifosfamide + E5-siRNA), indicating cell cycle arrest. The colony formation assay revealed a substantial decrease in the number of colonies following combination treatment. qRT-PCR analysis showed decreased expression of stemness-related genes CD44 and Nanog, and migration-related genes MMP2 and CXCL8 in the combination groups. Apoptosis-related genes Casp-3, Casp-9, and pP53 showed increased expression following combination therapy, while BAX expression increased and BCL2 expression decreased relative to the control. The study demonstrates that combining E5-siRNA with Oxaliplatin or Ifosfamide enhances the efficacy of chemotherapy in HPV-16 positive cervical cancer cells. This synergistic approach effectively targets multiple aspects of cancer cell behavior, including proliferation, apoptosis, migration, and stemness. The findings suggest that this combination strategy could potentially allow for lower chemotherapy doses, thereby reducing toxicity while maintaining therapeutic efficacy. This research provides valuable insights into targeting HPV E5 as a complementary approach to existing therapies focused on E6 and E7 oncoproteins, opening new avenues for combination therapies in cervical cancer treatment.

Cardiac fat adipocytes: An optimized protocol for isolation of ready-to-use mature adipocytes from human pericardial adipose tissue

A better understanding of the pathophysiology of cardiac fat depots is crucial to describe their role in the development of cardiovascular diseases. To this end, we have developed a method to isolate mature fat cells from the pericardial adipose tissue (PAT), the most accessible cardiac fat depot during cardiac surgery. Using enzymatic isolation, we were able to successfully obtain mature fat cells together with the corresponding cells of the stromal vascular fraction (SVF). We subjected the PAT adipocytes to thorough morphological and molecular characterization, including detailed fatty acid profiling, and simultaneously investigated their reactivity to external stimuli. Our approach resulted in highly purified fat cells with sustained viability for up to 72 h after explantation. Remarkably, these adipocytes responded to multiple challenges, including pro-inflammatory and metabolic stimuli, indicating their potential to trigger a pro-inflammatory response and modulate endothelial cell behavior. Furthermore, we have created conditions to maintain whole PAT in culture and preserve their viability and reactivity to external stimuli. The efficiency of cell recovery combined with minimal dedifferentiation underscores the promise for future applications as a personalized tool for screening and assessing individual patient responses to drugs and supplements or nutraceuticals.

Bone cells influence the degradation interface of pure Mg and WE43 materials: Insights from multimodal in vitro analysis

In this study, the interaction of pure Mg and WE43 alloy under the presence of osteoblast (OB) and osteoclast (OC) cells and their influence on the degradation of materials have been deeply analyzed. Since OB and OC interaction has an important role in bone remodeling, we examined the surface morphology and dynamic changes in the chemical composition and thickness of the corrosion layers formed on pure Mg and WE43 alloy by direct monoculture and coculture of pre-differentiated OB and OC cells in vitro. Electrochemical techniques examined the corrosion performance. The corrosion products were characterized using a combination of the focused ion beam (FIB), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). Cell viability and morphology were assessed by fluorescent microscopy and SEM. Our findings demonstrate cell spread and attachment variations, which differ depending on the Mg substrates. It was clearly shown that cell culture groups delayed degradation processes with the lowest corrosion rate observed in the presence of OBOC coculture for the WE43 substrate. Ca-P enrichment was observed in the outer-middle region of the corrosion layer but only after 7 days of OBOC coculture on WE43 and after 14 days on the pure Mg specimens. Statement of significanceMagnesium metallic materials that can degrade over time provide distinct opportunities for orthopedic application. However, there is still a lack, especially in elucidating cell-material interface characterization. This study investigated the influence of osteoblast-osteoclast coculture in direct Mg-material contact. Our findings demonstrated that pre-differentiated osteoblasts and osteoclasts cocultured on Mg substrates influenced the chemistry of the corrosion layers. The cell spread and attachment were Mg substrate-dependent. The findings of coculturing bone cells directly on Mg materials within an in vitro model provide an effective approach for studying the dynamic degradation processes of Mg alloys while also elucidating cell behavior and their potential contribution to the degradation of these alloys.

Multifaceted role of FAM210B in hepatocellular carcinoma: Implications for tumour progression, microenvironment modulation and therapeutic selection.

Hepatocellular carcinoma (HCC) is a common and lethal liver cancer characterized by complex aetiology and limited treatment options. FAM210B, implicated in various cancers, is noteworthy for its potential role in the progression and treatment response of HCC. Yet, its expression patterns, functional impacts and correlations with patient outcomes and resistance to therapy are not well understood. We employed a comprehensive methodology to explore the role of FAM210B in HCC, analysing its expression across cancers, subcellular localization and impacts on cell proliferation, invasion, migration, biological enrichment and the immune microenvironment. Additionally, we investigated its expression in single cells, drug sensitivity and relationships with genomic instability, immunotherapy efficacy and key immune checkpoints. While FAM210B expression varied across cancers, there was no notable difference between HCC and normal tissues. Elevated levels of FAM210B were associated with improved survival outcomes. Subcellular analysis located FAM210B in the plasma membrane and cytosol. FAM210B was generally downregulated in HCC, and its suppression significantly enhanced cell proliferation, invasion and migration. Biological enrichment analysis linked FAM210B to metabolic and immune response pathways. Moreover, its expression modified the immune microenvironment of HCC, affecting drug responsiveness and immunotherapy outcomes. High expression levels of FAM202B correlated with increased resistance to sunitinib and enhanced responsiveness to immunotherapy, as evidenced by associations with tumour mutation burden, PDCD1, CTLA4 and TIDE scores. FAM210B exerts a complex influence on HCC, affecting tumour cell behaviour, metabolic pathways, the immune microenvironment and responses to therapy.

Multiphoton excited polymerized biomimetic models of collagen fiber morphology to study single cell and collective migration dynamics in pancreatic cancer

The respective roles of aligned collagen fiber morphology found in the extracellular matrix (ECM) of pancreatic cancer patients and cellular migration dynamics have been gaining attention because of their connection with increased aggressive phenotypes and poor prognosis. To better understand how collagen fiber morphology influences cell-matrix interactions associated with metastasis, we used Second Harmonic Generation (SHG) images from patient biopsies with Pancreatic ductal adenocarcinoma (PDAC) as models to fabricate collagen scaffolds to investigate processes associated with motility. Using the PDAC BxPC-3 metastatic cell line, we investigated single and collective cell dynamics on scaffolds of varying collagen alignment. Collective or clustered cells grown on the scaffolds with the highest collagen fiber alignment had increased E-cadherin expression and larger focal adhesion sites compared to single cells, consistent with metastatic behavior. Analysis of single cell motility revealed that the dynamics were characterized by random walk on all substrates. However, examining collective motility over different time points showed that the migration was super-diffusive and enhanced on highly aligned fibers, whereas it was hindered and sub-diffusive on un-patterned substrates. This was further supported by the more elongated morphology observed in collectively migrating cells on aligned collagen fibers. Overall, this approach allows the decoupling of single and collective cell behavior as a function of collagen alignment and shows the relative importance of collective cell behavior as well as fiber morphology in PDAC metastasis. We suggest these scaffolds can be used for further investigations of PDAC cell biology. STATEMENT OF SIGNIFICANCE: Pancreatic ductal adenocarcinoma (PDAC) has a high mortality rate, where aligned collagen has been associated with poor prognosis. Biomimetic models representing this architecture are needed to understand complex cellular interactions. The SHG image-based models based on stromal collagen from human biopsies afford the measurements of cell morphology, cadherin and focal adhesion expression as well as detailed motility dynamics. Using a metastatic cell line, we decoupled the roles of single cell and collective cell behavior as well as that arising from aligned collagen. Our data suggests that metastatic characteristics are enhanced by increased collagen alignment and that collective cell behavior is more relevant to metastatic processes. These scaffolds provide new insight in this disease and can be a platform for further experiments such as testing drug efficacy.

On the Fused Deposition Modelling of Personalised Bio-Scaffolds: Materials, Design, and Manufacturing Aspects.

Bone tissue engineering (BTE) is an important field of research, essential in order to heal bone defects or replace impaired tissues and organs. As one of the most used additive manufacturing processes, 3D printing can produce biostructures in the field of tissue engineering for bones, orthopaedic tissues, and organs. Scaffold manufacturing techniques and suitable materials with final structural, mechanical properties, and the biological response of the implanted biomaterials are an essential part of BTE. In fact, the scaffold is an essential component for tissue engineering where cells can attach, proliferate, and differentiate to develop functional tissue. Fused deposition modelling (FDM) is commonly employed in the 3D printing of tissue-engineering scaffolds. Scaffolds must have a good architecture, considering the porosity, permeability, degradation, and healing capabilities. In fact, the architecture of a scaffold is crucial, influencing not only the physical and mechanical properties but also the cellular behaviours of mesenchymal stem cells. Cells placed on/or within the scaffolds is a standard approach in tissue engineering. For bio-scaffolds, materials that are biocompatible and biodegradable, and can support cell growth are the ones chosen. These include polymers like polylactic acid (PLA), polycaprolactone (PCL), and certain bioglass or composite materials. This work comprehensively integrates aspects related to the optimisation of biocompatible and biodegradable composites with the low cost, simple, and stable FDM technology to successfully prepare the best designed composite porous bone-healing scaffolds. FDM can be used to produce low-cost bone scaffolds, with a suitable porosity and permeability.

Design, development, and benchmarking of a bioreactor integrated with 3D bioprinting: Application to skeletal muscle regeneration

In recent years, great efforts have been spent to create engineered muscle constructs recapitulating the 3D architecture and applying external stimulations. In this regard, tissue engineering approaches could be very promising in regenerating skeletal muscle, in which bioprinting techniques have produced encouraging results especially regarding 3D architecture. Tensile stimuli showed a fundamental role in regulating the behavior of muscle cells both in terms of 3D organizations and protein expression. Despite this promising premise, the combination of 3D bioprinting and mechanical stimulation has been poorly investigated, calling for novel approaches dealing with the mechanical stimulation of the 3D bioprinted construct and the integration of the bioprinting phase into the stimulation device. To this aim, the present work proposes the design, manufacturing, and benchmarking of a bioprinting-integrated mechanical platform conceived for mechanically stimulating a 3D muscle model directly printed into the bioreactor to foster the integration of printing and stimulation. The study consists of three main steps: 1) the design, fabrication, and mechanical characterization of stretchable supports suitable for bioprinting and long-term cell culture; 2) the design, assisted by computational tools, and the fabrication of the smart Petri dish containing the stimulation mechanism and of the final cyclic mechanical platform; 3) the in-vitro validation of the proposed platform in terms of transmission of the mechanical stimulation to the 3D construct and the biological effect of dynamic culture on 3D bioprinted muscle cells. The results highlighted excellent viability and demonstrated that the external stimulus influences the murine myoblasts behavior already after 7 days of culture. In conclusion, prototypes are now available of a mechanical platform that integrates the 3D bioprinting and is capable of stimulating 3D biological constructs for applications in the field of muscle tissue engineering.

Extracellular Matrix Sulfation in the Tumor Microenvironment Stimulates Cancer Stemness and Invasiveness.

Tumor extracellular matrices (ECM) exhibit aberrant changes in composition and mechanics compared to normal tissues. Proteoglycans (PG) are vital regulators of cellular signaling in the ECM with the ability to modulate receptor tyrosine kinase (RTK) activation via their sulfated glycosaminoglycan (sGAG) side chains. However, their role on tumor cell behavior is controversial. Here, it is demonstrated that PGs are heavily expressed in lung adenocarcinoma (LUAD) patients in correlation with invasive phenotype and poor prognosis. A bioengineered human lung tumor model that recapitulates the increase of sGAGs in tumors in an organotypic matrix with independent control of stiffness, viscoelasticity, ligand density, and porosity, is developed. This model reveals that increased sulfation stimulates extensive proliferation, epithelial-mesenchymal transition (EMT), and stemness in cancer cells. The focal adhesion kinase (FAK)-phosphatidylinositol 3-kinase (PI3K) signaling axis is identified as a mediator of sulfation-induced molecular changes in cells upon activation of a distinct set of RTKs within tumor-mimetic hydrogels. The study shows that the transcriptomic landscape of tumor cells in response to increased sulfation resembles native PG-rich patient tumors by employing integrative omics and network modeling approaches.

The impact of valproic acid administration: Effects on the growth of tongue cancer cells

Background: Tongue cancer represents the predominant malignancy within the oral cavity (25-40% of squamous cell carcinoma), necessitating treatment modalities such as surgery, radiotherapy, and chemotherapy. Valproic acid, an antiepileptic medication, functions as a histone deacetylase inhibitor or activator of anti-tumor signaling pathways. Objective: To deepen our understanding of the effects of valproic acid on the viability, cytotoxicity, proliferation, and migration capabilities of HSC-3 cells. Method: This study employed an in vitro laboratory approach, exposing HSC-3 cells to valproic acid. The experimental groups included a negative control with culture media devoid of valproic acid, and treatment groups exposed to valproic acid at concentrations of 145 ppm, 180 ppm, and 355 ppm, respectively. Results: Significant differences (p-value <0.05) were observed between HSC-3 cells treated with valproic acid (145 ppm, 180 ppm, and 355 ppm) and the control group in terms of viability, cytotoxicity, proliferation, and migration. Reduced cell viability, increased cytotoxicity, and decreased proliferation were noted. Migration assays indicated suppressed migration of HSC-3 cells. Conclusion: In summary, this study reveals that valproic acid exerts substantial effects on various aspects of HSC-3 cell behavior. It decreases cell viability, enhances cytotoxicity, suppresses proliferation, and inhibits cell migration. These findings highlight the potential of valproic acid as a therapeutic agent for tongue cancer by targeting crucial cellular processes involved in cancer progression. Further research and clinical trials are essential to confirm these effects and explore their application in cancer treatment strategies. Novelty/Originality of this article:: This study shows valproic acid has potential as a therapeutic agent for tongue cancer by decreasing cell viability, increasing cytotoxicity, suppressing proliferation, and inhibiting migration of HSC-3 cells. These findings introduce a new application of valproic acid as an anticancer agent, expanding the use of antiepileptic drugs. This study opens up opportunities for developing more effective tongue cancer therapies and encourages further research and clinical trials to validate these findings

A novel modified complementary metamaterial resonator based dual-band bandpass quasi-elliptic filter using half-mode SIW cavity with wide stopband rejection for wireless communication applications

This article presents a novel dual-band metamaterial bandpass quasi-elliptic filter (DBBPF) for wireless communication applications, addressing the critical need for compact, high-performance multi-band filters in modern systems. The proposed filter leverages substrate-integrated waveguide cavity (SIWC) technology combined with innovative complementary metamaterial resonators to achieve significant miniaturization while maintaining excellent electrical performance. The filter’s design incorporates four complementary resonators of modified rectangular shape (CMSRR) on the metalized top face, generating targeted operating bands below the SIW cutoff frequency. To control their resonances, the size of each CMSRR is optimized for electrical dimensions of just 0.154 λ0× 0.115 λ0× 0.028 λ0. We present a comprehensive analysis of both full mode (FMSIWC) and half mode (HMSIWC) configurations, demonstrating dual-bandpass behavior with central frequencies at 5.79 and 9.82 GHz for FMSIWC, and 5.72 and 9.74 GHz for HMSIWC. The electromagnetic behavior of the basic cell of the filter is analyzed based on the frequency characteristics of its permittivity and permeability. A parametric study according to the location of the metamaterial resonators in the filter is conducted to have the optimized dimensions. Additionally, the confinement of the electric field in the two filter configurations is discussed to better understand their behavior. The fabricated HMSIWC filter, implemented on an FR4-Epoxy substrate measuring only 54.60 × 25.75 × 1.5875 mm3, achieves remarkable miniaturization while maintaining desired performance characteristics. Experimental results validate the simulated performance, demonstrating excellent agreement for both configurations. Key performance metrics include fractional bandwidths of 3.77 and 6.98 %, and insertion losses of 1.33 dB and 2.14 dB for the two passbands in the HMSIWC design. This work advances the state-of-the-art SIW filter design by combining half-mode techniques with optimized CMSRR structures, resulting in a compact, high-performance dual-band filter. The proposed DBBPF’s simple design and small form factor make it an ideal candidate for integration into various wireless communication systems, particularly where space constraints are critical.

Inhibition of DDR1 promotes ferroptosis and overcomes gefitinib resistance in non-small cell lung cancer

Gefitinib is an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI), which serves the critical pillar for the treatment of non-small cell lung cancer (NSCLC). However, the acquired resistance remains a challenge for its clinical application, for which, practical strategies to reverse gefitinib resistance in NSCLC are necessary. Ferroptosis, a programmed cell death driven by ferritin-dependent lipid peroxidation, involves in NSCLC progression and related chemoresistance. In our previous work, the self-synthesised EGFR inhibitor Yfq07 (N4, N6-disubstituted pyrimidine-4,6-diamine derivatives) displayed a considerable inhibitory effect on NSCLC both in vitro and in vivo. Herein, we observed that Yfq07 suppressed the proliferation of PC-9GR and HCC827GR cells, two gefitinib resistance NSCLC cell lines. Mechanically, Yfq07 inhibited the phosphorylation of the Discoidin Domain Receptor 1 (DDR1), a receptor tyrosine kinase (RTK) highly expressed in multiple cancers, accompanied by downregulated miR-3648 and upregulated SOCS2. Inhibition or knockdown of DDR1 suppressed the proliferation, migration, and invasion of gefitinib-resistant NSCLC cells, and on the other hand, also downregulated miR-3648 and promoted SOCS2 expression. More specifically, miR-3648 targeted the 3’UTR segment of SOCS2 mRNA and thus affecting the P-ERK signalling pathway to regulate the malignant behaviors of gefitinib-resistant NSCLC cells. Furthermore, Yfq07 also indirectly induced the ferroptosis of gefitinib-resistant NSCLC cells via SOCS2 triggered inhibition of xCT-GPX4 pathway. In conclusion, our study indicates that DDR1 inhibitor Yfq07 promotes ferroptosis and reverses gefitinib-resistance of NSCLC through DDR1-miR-3648-SOCS2 signalling pathway, which provides insights for targeted therapy of gefitinib-resistant NSCLC and drug developments targeting ferroptosis.

Mechanical Properties and Deformation Behavior of Open-Cell Type Aluminum Foams with Structural Conditions and Alloy Composition

Open-cell type aluminum foams with various structural conditions and alloy compositions were manufactured using the replication casting process. The porosity of the foams ranged from 55% to 62%, with pore sizes of 0.7~1.0 mm, 1.0~2.0 mm, and 2.8~3.4 mm. The alloys employed included commercial A356 and A383, as well as Al-6Mg-(0, 2, 4, 6)Si alloys. Compression tests were conducted under various conditions of the foams, and the results were comparatively analyzed based on the detailed structural conditions and alloy compositions. It was observed that for the same alloy composition and equivalent porosity, a reduction in pore size led to an increased number of cell walls, enhancing energy dispersion and resulting in higher compressive yield strength and energy absorption. Under the same pore size, a decrease in porosity increased the relative density and cell wall thickness, leading to improved compressive yield strength and energy absorption. Furthermore, compressive evaluation based on alloy composition revealed the influence of the inherent mechanical properties of the material on the mechanical properties of open-cell type aluminum foams. Specifically, it was confirmed that alloys with higher ductility exhibited elastic behavior of the internal cells under external stress, significantly influencing the energy absorption of foams.

Integrated overview of stramenopile ecology, taxonomy, and heterotrophic origin.

Stramenopiles represent a significant proportion of aquatic and terrestrial biota. Most biologists can name a few, but these are limited to the phototrophic (e.g., diatoms and kelp) or parasitic species (e.g., oomycetes, Blastocystis), with free-living heterotrophs largely overlooked. Though our attention is slowly turning towards heterotrophs, we have only a limited understanding of their biology due to a lack of cultured models. Recent metagenomic and single-cell investigations have revealed the species richness and ecological importance of stramenopiles-especially heterotrophs. However, our lack of knowledge of the cell biology and behaviour of these organisms leads to our inability to match species to their particular ecological functions. Because photosynthetic stramenopiles are studied independently of their heterotrophic relatives, they are often treated separately in the literature. Here, we present stramenopiles as a unified group with shared synapomorphies and evolutionary history. We introduce the main lineages, describe their important biological and ecological traits, and provide a concise update on the origin of the ochrophyte plastid. We highlight the crucial role of heterotrophs and mixotrophs in our understanding of stramenopiles with the goal of inspiring future investigations in taxonomy and life history. To understand each of the many diversifications within stramenopiles-towards autotrophy, osmotrophy, or parasitism-we must understand the ancestral heterotrophic flagellate from which they each evolved. We hope the following will serve as a primer for new stramenopile researchers or as an integrative refresher to those already in the field.