Junction Formation Research Articles

Evaluating Physiological and Organizational Properties of the Tight Junction ZO-1 ZU-5 Domain in MDCK Cells

Zonula occludens-1 (ZO-1) is an essential TJ component contributing to junctional formation by mediating interactions between junctional proteins and the actomyosin cytoskeleton. MDCK II cells, a model epithelial cell line, with ZO-1 knocked-out (ZO-1 KO) have observable phenotypic changes such as alterations in the actin cytoskeletal organization and physiological barrier properties. This study focuses on the ZO-1-ZU5 domain by rescuing ZO-1 with full-length and a panel of ZO-1-ZU5 mutants. ZO-1-ZU-5 is known to bind cingulin and through this interaction ZO-1 adopts an extended conformation. We examined actomyosin cytoskeleton organization, ZO-1 mobility, gene expression and barrier function to determine whether the ZO-1-ZU5 mutants rescue full-length activity. We found that the ZO-1- ZU5 truncation mutant cell line rescued transepithelial electrical resistance (TER) but did not rescue tight junction distribution or the amount of total gamma-actin. We will present evaluation of additional ZU5 mutants. This study expands our understanding of how extended ZO-1 regulates the mechanical, genetic, and phenotypic properties of epithelial cells. Trinity University Biology Summer Undergraduate Research Fellowship. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Investigation of the Role of Aging on Pulmonary Microvascular Endothelial Cell Barrier Function During Mechanical Ventilation

Elderly individuals have substantially elevated morbidity and mortality following pulmonary insult that require mechanical ventilation (MV) to sustain respiratory function. It is not known if an altered response to MV contributes to the worsened outcomes in the elderly. MV is associated with injury to pulmonary microvascular endothelial cells (PMVEC), leading to a compromised vascular barrier, and fluid and protein leakage within the tissue. However, the precise impact that age has on the vascular endothelium during MV is unknown. We hypothesized that aging augments PMVEC barrier dysfunction during MV. To address this hypothesis, young and aged mice were ventilated at a tidal volume of 20 mL/kg for 3 hours. To assess microvascular permeability, lungs were lavaged and abundance of the serum protein, immunoglobulin M, within the lavage fluid was measured. Lung tissue was dissociated into single cells and RNA was collected for single-cell RNA sequencing (scRNAseq). Subsequent analysis was performed using the Seurat package in R to give insight into the role of specific cell types, with a particular emphasis on microvascular endothelial cells. To directly assess molecular mechanisms directly in the cells, PMVEC were isolated from a separate cohort of non-ventilated young and aged mice and cultured in vitro. Cells were grown to confluency, then assessed for barrier integrity by Evans blue-labelled albumin leak across monolayers and immunofluorescence staining of cell junctional adherens and tight junction proteins, VE-cadherin and claudin-5. Compared to young, aged-ventilated animals had a significant increase in immunoglobulin M in their lavage fluid. Analysis of scRNAseq data revealed more robust activation and inflammation in the endothelial cells from aged animals following MV. Endothelial monolayers from aged animals in vitro exhibited a significant increase in albumin flux, which was associated with poor VE-cadherin and claudin-5 junction formation. In conclusion, PMVEC from aged mice exhibit compromised barrier function, leading to augmented permeability following MV. The age-dependent loss in barrier function appears to be due to an impaired ability to re-establish functional cell-cell junctions. These findings suggest that the aged vascular barrier is more susceptible to injury from MV, due to impaired PMVEC cell-cell junction integrity. Our results may highlight molecular pathways involved in predisposing aged individuals to worsened outcomes during MV, which can help in developing targeted therapeutics. Funded by Lawson Health Research Institute, CIHR, the Ontario Graduate Scholarship, and Western University. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Disaggregation and fibrillation during sol-gel transition of alginate hydrogels

The rheological and morphological characteristics of Ca-crosslinked alginate hydrogels with two different M/G ratios, α-L-guluronate (G)-rich and β-D-mannuronate (M)-rich, each with one alginic acid concentration, were investigated. It was found that the stiffness and elasticity of alginate hydrogels are derived from the thickness and density of the fibril network structures. In aqueous alginate solution, ball-like aggregates of alginates are present. Time-resolved small-angle X-ray scattering and time-domain nuclear magnetic resonance measurements suggest that the disaggregation of alginate aggregates and loose fibrillation occur in the early stage of the sol-gel transition. After these induction stage, direct gelation is finally caused by the formation of the egg-box junction. G-rich alginate hydrogel has a higher stiffness and a thicker and denser fibril network structure than M-rich alginate hydrogel. The former also exhibits faster and more significant changes in physical properties during the sol-gel transition.

Lactiplantibacillus plantarum IS-10506 Enhances Tight Junction Integrity in Bronchial Epithelium: An Experimental Study

BACKGROUND: Airway epithelium constitutes the initial structural defense against inhaled allergens, with its integrity primarily maintained through the formation of tight junctions (TJ) consisting of claudins, occludins, and zonula occludens (ZO). Currently, the effects of probiotics on airway TJ remain unclear. Therefore, this study was conducted to explore the impact of Lactiplantibacillus plantarum IS-10506, a native probiotic from Indonesia, on bronchial epithelial TJ in a mouse model of allergic airway inflammation.METHODS: Twenty-four BALB/c mice were randomly assigned to three groups: negative control, positive control subjected to Dermatophagoides pteryonissinus (Der p) sensitization and challenge, and treatment group exposed to Der p and administered L. plantarum IS-10506 since one week before sensitization up to three weeks after. The expression levels of claudin-18, occludin, and ZO-1 were investigated through immunohistochemistry analysis and quantified based on the immunoreactive score (IRS). RESULTS: The results of this study demonstrated that L. plantarum IS-10506 significantly enhanced the expressions of claudin-18, occludin, and ZO-1 (p<0.05).CONCLUSION: These findings underscore the probiotic's role in fortifying the integrity of airway TJ.KEYWORDS: allergic inflammation, probiotic Lactiplantibacillus plantarum, immunology, respiratory system, tight junction, occludin, claudin, zonula occludens

Synthesis of the composites of carbon quantum dots and metal organic framework and their application in hydrogen evolution

The Ti-MIL-125 metal-organic framework (MOF) functions as a nanoreactor, producing carbon quantum dots (CQDs) within its structure and forming CQD/MOF composites. Similarly, a Pt-CQD/MOF three-component composite is synthesized. Their properties, along with those of CQDs and MOF-free-CQDs (mf-CQDs) enclosed within the MOF, undergo assessment and comparison. Integrating CQDs and Pt minimally alters the MOF structure but reduces specific surface areas due to incorporation. The emitted wavelength of CQD/MOF (570 nm) differs from pristine CQDs (450 nm) and mf-CQDs (496 nm), indicating MOF influence on CQDs' growth and photoluminescence. HOMO and LUMO levels of CQDs/MOF, measured via ultraviolet photoelectron spectroscopy and low-energy inverse photoelectron spectroscopy, are more positive than those of the MOF, suggesting heterogeneous junction formation, aiding electron separation. CQD/MOF exhibits a modest photocatalytic hydrogen evolution rate of 20.4 μmol/g/h. Conversely, Pt-CQD/MOF significantly boosts the rate to 2213 μmol/g/h, underscoring Pt's pivotal catalytic role.

Cryopreservation and Rapid Recovery of Differentiated Intestinal Epithelial Barrier Cells at Complex Transwell Interfaces Is Enabled by Chemically Induced Ice Nucleation.

Cell-based models, such as organ-on-chips, can replace and inform in vivo (animal) studies for drug discovery, toxicology, and biomedical science, but most cannot be banked "ready to use" as they do not survive conventional cryopreservation with DMSO alone. Here, we demonstrate how macromolecular ice nucleators enable the successful cryopreservation of epithelial intestinal models supported upon the interface of transwells, allowing recovery of function in just 7 days post-thaw directly from the freezer, compared to 21 days from conventional suspension cryopreservation. Caco-2 cells and Caco-2/HT29-MTX cocultures are cryopreserved on transwell inserts, with chemically induced ice nucleation at warmer temperatures resulting in increased cell viability but crucially retaining the complex cellular adhesion on the transwell insert interfaces, which other cryoprotectants do not. Trans-epithelial electrical resistance measurements, confocal microscopy, histology, and whole-cell proteomics demonstrated the rapid recovery of differentiated cell function, including the formation of tight junctions. Lucifer yellow permeability assays confirmed that the barrier functions of the cells were intact. This work will help solve the long-standing problem of transwell tissue barrier model storage, facilitating access to advanced predictive cellular models. This is underpinned by precise control of the nucleation temperature, addressing a crucial biophysical mode of damage.

Cascaded autocatalytic hairpin assembly molecular circuit for amplified fluorescent aptamer luteinising hormone assay

The quantitative detection of luteinising hormone (LH) is critical for the study of the physiological mechanism of reproductive function and the assessment of infertility and the clinical treatment of reproductive disorders. However, conventional approaches for LH detection are mostly based on an antibody recognition module with the limitations of sensitivity, simplicity and cost. The development of robust LH sensing methods is therefore highly demanded for facilitating the diagnosis of LH-related diseases. We establish a convenient, amplified and sensitive fluorescent aptamer LH assay based on new target-triggered and cascaded autocatalytic hairpin assembly (C-aCHA) circuit amplification means via initiator sequence replication. Target LH molecules bind the aptamers in the aptamer/initiator duplexes to release the initiator sequences, which trigger CHA formation of DNA three-way junctions (TWJs) and the unfolding of fluorescently quenched signal hairpins to show amplified fluorescence. The TWJs further activate another CHA cycle for the yield of more initiator sequences to form the C-aCHA circuit amplification cycles, which lead to the unfolding of many signal hairpins to exhibit substantially magnified fluorescence recovery for detecting LH down to 8.56 pM in the range from 10 pM to 50 nM. In addition, the monitoring of trace LH in diluted serums by this sensing approach has been also verified. Our LH assay clearly outperforms current existing antibody-based methods and the C-aCHA signal amplification strategy can be easily extended as a robust means for sensitively monitoring various biomolecular markers with simple replacement of the corresponding aptamers for diverse applications.

Claudins in Cancer: A Current and Future Therapeutic Target.

Claudins are a family of 27 proteins that have an important role in the formation of tight junctions. They also have an important function in ion exchange, cell mobility, and the epithelial-to-mesenchymal transition, the latter being very important in cancer invasion and metastasis. Therapeutic targeting of claudins has been investigated to improve cancer outcomes. Recent evidence shows improved outcomes when combining monoclonal antibodies against claudin 18.2 with chemotherapy for patients with gastroesophageal junction cancer. Currently, chimeric antigen receptor T-cells targeting claudin 18 are under investigation. In this review, we will discuss the major functions of claudins, their distribution in the normal as well as cancerous tissues, and their effect in cancer metastasis, with a special focus on the therapeutic targeting of claudins to improve cancer outcomes.

NiAlFe LTH /MoS2 p-n junction heterostructure composite as an effective visible-light-driven photocatalyst for enhanced degradation of organic dye under high alkaline conditions

Designing of an effectual heterostructure photocatalyst for catalytic organic pollutant exclusion has been the subject of rigorous research intended to resolve the related environmental aggravation. Fabricating p-n junctions is an effective strategy to promote electron-hole separation of semiconductor photocatalysts as well as enhance the organic toxin degradation performance. In this study, a series of n-type NiAlFe-layered triple hydroxide (LTH) loaded with various ratios of p-type MoS2 was synthesized for forming a heterostructure LTH/MoS2 (LMs) by an in situ hydrothermal strategy. The photocatalysts were characterized by XRD, SEM&EDX, TEM, FT-IR, XPS, as well as UV–vis DRS. The photoactivity of photocatalysts was tested by the degradation of Indigo Carmine (IC) dye. The optimized catalyst (LM1) degrades 100% of indigo dye in high alkaline pH under UV light for 100 min. Besides, the degradation rate of LM1 is 15 times higher than that of pristine NiAlFe-LTH. The enhanced photoactivity is attributed to the synergistic effect between NiAlFe-LTH and MoS2 as well as the p-n junction formation.

Laser shock-enabled optical–thermal–mechanical coupled welding method for silver nanowires

Silver nanowires (AgNWs) are recognized as highly promising materials for flexible and transparent electrode applications. However, existing material-processing methods fail to achieve uniform and reliable AgNWs junctions. In this study, we propose a new method using the laser shock effect combined with the laser heating effect, for creating AgNW junctions within thin films. We explored the welding mechanism of AgNWs through optic-thermal welding, laser shock-enabled mechanical welding, and laser-shock-enabled optical-thermal-mechanical (LS-OTM) experiments, as well as numerical simulations, and the results demonstrate that the innovative mechanism of the LS-OTM process lies in its utilization of laser shock to adjust the gap between the nanowire junctions, which in turn achieves a fine control of the thermal effect of the heating laser localised surface plasmon resonance, and the atomic diffusion in the solid state at intermediate temperature under the action of the impact force is the mechanism of the formation of high-quality junctions. We prepared flexible transparent conductive films and studied their transmittance, conductivity, and thermal properties, the results show that the flexible transparent conductive films prepared by LS-OTM welding method have excellent transmittance, conductivity, and thermal properties, this verifies the feasibility and effectiveness of this processing strategy. The LS-OTM method is a viable solution for manufacturing transparent, conductive films from AgNWs for emerging applications such as flexible heated films, flexible displays, and wearable medical devices.

The DEAD-box ATPase Dbp10/DDX54 initiates peptidyl transferase center formation during 60S ribosome biogenesis

DEAD-box ATPases play crucial roles in guiding rRNA restructuring events during the biogenesis of large (60S) ribosomal subunits, but their precise molecular functions are currently unknown. In this study, we present cryo-EM reconstructions of nucleolar pre-60S intermediates that reveal an unexpected, alternate secondary structure within the nascent peptidyl-transferase-center (PTC). Our analysis of three sequential nucleolar pre-60S intermediates reveals that the DEAD-box ATPase Dbp10/DDX54 remodels this alternate base pairing and enables the formation of the rRNA junction that anchors the mature form of the universally conserved PTC A-loop. Post-catalysis, Dbp10 captures rRNA helix H61, initiating the concerted exchange of biogenesis factors during late nucleolar 60S maturation. Our findings show that Dbp10 activity is essential for the formation of the ribosome active site and reveal how this function is integrated with subsequent assembly steps to drive the biogenesis of the large ribosomal subunit.

Surface decoration of flexible Si3N4 nanowire membrane by hydroxyapatite micron-flake with excellent thermal insulation at 1300 °C

Flexible and excellent thermal insulation membranes are required as the thermal protection system materials of aerospace vehicles for next-generation space missions. Using ceramic membranes with ultralow thermal conductivity has been proved to be an effective way to protect the electronic components inside the fuselage. Herein, we decorate hydroxyapatite (HA) micron-flake onto Si3N4 nanowire membrane (SR), and investigate the thermal stability, fire-resistance and thermal insulating properties of the obtained SR decorated by HA micron-flake (SRHM). Surface decoration of HA micron-flake onto the SR results in the formation of well-interconnected junctions and the increase of interfacial thermal resistance. Due to the increased interfacial thermal resistance, the as-prepared SRHM exhibits good thermal insulation and outstanding fire-retardant performance. SRHM shows robust mechanical property with tensile strength value of 4.10 MPa, increased by 247.5% compared to the SR without the surface decoration by HA micron-flake. The thermal conductivity of SRHM is as low as 0.037 W m−1 K−1, reduced by 33.9% compared to the SR without the surface decoration by HA micron-flake. After SRHM is heated by a butane blow torch at 1300 °C for 30 min, no obvious change is observed in the macroscopic morphology. And the temperature of the backside for SRHM maintains at a stable temperature of ∼260 °C for 300 s, which is 1040 °C lower than that of the front side. Additionally, SRHM can protect the electronic components form being burnt under the heating of an alcohol lamp for 30 min. The successful preparation of such thermal insulating and fire-retardant SRHM will open up a new world for the widespread applications of ceramic membranes in the fields of aerospace vehicles.

Enhanced selectivity of novel sea urchin-like h-/m-WO3 hetero nanoflowers for highly sensitive detection of ammonia by double filtration method

Real-time monitoring of toxic gases and breath markers can be performed by metal oxide semiconductor-based chemo-resistive gas sensors due to their simple structures, low cost, and faster and better recovery and response. Due to the effective electron-hole separation, tuning the development of phase junctions of the metal oxide is an efficient method for improving their gas sensing properties. In this study, two distinct hydrothermal methods were used to synthesize h-/m-WO3 hetero-nanoflowers. In the first method, L-Cysteine was used to facilitate the formation of phase junctions between h-WO3 and m-WO3 nanostructures, whereas Thiourea was used in the second. Various characterisation approaches validated the hexagonal-monoclinic phase junction development in the nanoflowers. The sensing material fabricated using Thiourea with a concentration of 0.01M performed better than other sensors. At 350°C, the selected sample displayed outstanding ammonia sensing capacity with the experimental detection limit of 1 ppm (response=3.33). The theoretical detection limit is determined to be 27 ppb. The selectivity of the sensor towards NH3 over H2S and acetone is increased by combining the devices with a double filtration setup. The effectiveness of the selected h-/m-WO3 sensor for diagnosing kidney dysfunction was evaluated by analyzing simulated breath samples.

An engineered in vitro model of the human myotendinous junction

The myotendinous junction (MTJ) is a vulnerable region at the interface of skeletal muscle and tendon that forms an integrated mechanical unit. This study presents a technique for the spatially restrictive co-culture of human embryonic stem cell (hESC)-derived skeletal myocytes and primary tenocytes for two-dimensional modeling of the MTJ. Micropatterned lanes of extracellular matrix and a 2-well culture chamber define the initial regions of occupation. On day 1, both lines occupy less than 20 % of the initially vacant interstitial zone, referred to henceforth as the junction. Myocyte–tenocyte interdigitations are observed by day 7. Immunocytochemistry reveals enhanced organization and alignment of patterned myocyte and tenocyte features, as well as differential expression of multiple MTJ markers. On day 24, electrically stimulated junction myocytes demonstrate negative contractile strains, while positive tensile strains are exhibited by mechanically passive tenocytes at the junction. Unpatterned tenocytes distal to the junction experience significantly decreased strains in comparison to cells at the interface. Unpatterned myocytes have impaired organization and uncoordinated contractile behavior. These findings suggest that this platform is capable of inducing myocyte–tenocyte junction formation and mechanical coupling similar to the native MTJ, showing transduction of force across the cell–cell interface. Statement of significanceThe myotendinous junction (MTJ) is an integrated structure that transduces force across the muscle-tendon boundary, making the region vulnerable to strain injury. Despite the clinical relevance, previous in vitro models of the MTJ lack the structure and mechanical accuracy of the native tissue and have difficulty transmitting force across the cell–cell interface. This study demonstrates an in vitro model of the MTJ, using spatially restrictive cues to inform human myocyte–tenocyte interactions and architecture. The model expressed MTJ markers and developed anisotropic myocyte–tenocyte integrations that resemble the native tissue and allow for force transduction from contracting myocytes to passive tenocyte regions. As such, this study presents a system capable of investigating development, injury, and pathology in the human MTJ.

Bacterial Biohybrids for Invasion of Tumor Cells Promote Antigen Cross-Presentation Through Gap Junction.

Due to inherent differences in cellular composition and metabolic behavior with host cells, tumor-harbored bacteria can discriminatorily affect tumor immune landscape. However, the mechanisms by which intracellular bacteria affect antigen presentation process between tumor cells and antigen-presenting cells (APCs) are largely unknown. The invasion behavior of attenuated Salmonella VNP20009 (VNP) into tumor cells is investigated and an attempt is made to modulate this behavior by modifying positively charged polymers on the surface of VNP. It is found that non-toxic chitosan oligosaccharide (COS) modified VNP (VNP@COS) bolsters the formation of gap junction between tumor cells and APCs by enhancing the ability of VNP to infect tumor cells. On this basis, a bacterial biohybrid is designed to promote in situ antigen cross-presentation through intracellular bacteria induced gap junction. This bacterial biohybrid also enhances the expression of major histocompatibility complex class I molecules on the surface of tumor cells through the incorporation of Mdivi-1 coupled with VNP@COS. This strategic integration serves to heighten the immunogenic exposure of tumor antigens; while, preserving the cytotoxic potency of T cells. A strategy is proposed to precisely controlling the function and local effects of microorganisms within tumors.

Abstract LB057: Pre-clinical characterization of IBI343, a site-specifically conjugated anti-Claundin18.2 ADC, for treating solid tumors

Abstract Claudin18.2 is important for the formation and function of tight junctions between cells in epithelial tissues. Normally expressed only in gastric mucosa, Claudin18.2 is overexpressed in multiple types of solid tumors, including gastric, pancreatic, breast and colon cancers. Despite being a well-established tumor-associated antigen, the clinical value and feasibility of targeting Claudin18.2 by various modalities are yet-to-be fully demonstrated. IBI343 is a fully humanized anti-Claudin18.2 monoclonal antibody conjugated to Exatecan via site-specific glycol-conjugation with an average drug-to-antibody ratio (DAR) of 3.6. IBI343 showed robust antigen-specific in vitro cytotoxicity in cancer cell lines with varying levels of Claudin18.2 expression. In addition, due to the cleavable linker design and high hydrophilicity of the payload, IBI343 demonstrated potent bystander killing effects in vitro. The glycan-based conjugation technology leads to enhanced stability of the conjugated linker-payload and the entire ADC molecule, which results in superior pharmacokinetics. IBI343 showed potent in vivo anti-tumor efficacy with complete tumor regressions in multiple xenograft models. Furthermore, it demonstrated strong in vitro and in vivo synergistic effects with various anti-cancer agents, including immune checkpoint blockers and DNA damage repair pathway inhibitors. IBI343 displayed good safety profile in monkey GLP toxicology study (HNSTD = 30 mg/kg). These pre-clinical findings show that IBI343 is a promising anti-cancer agent with large therapeutic window, and support clinical exploration of IBI343 in patients with Claudin18.2-expressing tumors. A companion late-breaking abstract of clinical phase 1 study results (NCT05458219) will also be presented in AACR 2024. Citation Format: Shuaixiang Zhou, Xiong Yao, Jian Guan, Keke Fei, Jia Lu, Weiwei Wu, Yang Liu, Tianyu Zhu, Zhuangguang Liao, Shi Chen, Bingliang Chen, Kaijie He. Pre-clinical characterization of IBI343, a site-specifically conjugated anti-Claundin18.2 ADC, for treating solid tumors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(7_Suppl):Abstract nr LB057.

Amorphous and crystalline heterostructure MoSe2/NiFe-LDH through p-n junction formation for electrochemical water splitting-synergistic back bonding effect

Designing a long-lasting, highly effective, non-noble metal-free electrocatalyst for both hydrogen evolution reaction (HER) and the oxygen evolution reaction (OER) is a strenuous task for water electrolysis. Here, in our study, an ordered NiFe-layer double hydroxide (LDH)/MoSe2 as p-n junction material was designed using facile two-step hydrothermal process for the overall water splitting reaction. Additionally, synthesized NiFe-LDH/MoSe2, electrode exhibits long-term durability in both acidic and alkaline medium for HER and OER, respectively. Mott-Schottky experiments provides the insights of interfacial electron transfer, where the electron is transported from n-type NiFe-LDH to p-type MoSe2, resulting in the generation of the p-n junction in MoSe2/NiFe-LDH. As the formation LDH and transition dichalcogenides p-n junction leads to activation of the catalysts towards the OH− adsorption that may enable the water electrolysis with a low overpotential. The XPS analysis supports the electron transfer from Fe to Mo resulting in the formation synergistic back bonding between the p-n heterojunction. This strategy opens up an alternative path to a highly effective, relatively low-price catalyst for the global production of renewable energy fuels.

MicroRNA Regulatory Pattern in Diabetic Mouse Cortex at Different Stages Following Ischemic Stroke.

After ischemic stroke, microRNAs (miRNAs) participate in various processes, including immune responses, inflammation, and angiogenesis. Diabetes is a key factor increasing the risk of ischemic stroke; however, the regulatory pattern of miRNAs at different stages of diabetic stroke remains unclear. This study comprehensively analyzed the miRNA expression profiles in diabetic mice at 1, 3, and 7 days post-reperfusion following the middle cerebral artery occlusion (MCAO). We identified differentially expressed (DE) miRNAs in diabetic stroke and found significant dysregulation of some novel miRNAs (novel_mir310, novel_mir89, and novel_mir396) post-stroke. These DEmiRNAs were involved in apoptosis and the formation of tight junctions. Finally, we identified three groups of time-dependent DE miRNAs (miR-6240, miR-135b-3p, and miR-672-5p). These have the potential to serve as biomarkers of diabetic stroke. These findings provide a new perspective for future research, emphasizing the dynamic changes in miRNA expression after diabetic stroke and offering potential candidates as biomarkers for future clinical applications.

Molecular and Cellular Characterization of Primary Endothelial Cells from a Familial Cavernomatosis Patient.

Cerebral cavernous malformation (CCM) or familial cavernomatosis is a rare, autosomal dominant, inherited disease characterized by the presence of vascular malformations consisting of blood vessels with an abnormal structure in the form of clusters. Based on the altered gene (CCM1/Krit1, CCM2, CCM3) and its origin (spontaneous or familial), different types of this disease can be found. In this work we have isolated and cultivated primary endothelial cells (ECs) from peripheral blood of a type 1 CCM patient. Differential functional and gene expression profiles of these cells were analyzed and compared to primary ECs from a healthy donor. The mutation of the familial index case consisted of a heterozygous point mutation in the position +1 splicing consensus between exons 15 and 16, causing failure in RNA processing and in the final protein. Furthermore, gene expression analysis by quantitative PCR revealed a decreased expression of genes involved in intercellular junction formation, angiogenesis, and vascular homeostasis. Cell biology analysis showed that CCM1 ECs were impaired in angiogenesis and cell migration. Taken together, the results obtained suggest that the alterations found in CCM1 ECs are already present in the heterozygous condition, suffering from vascular impairment and somewhat predisposed to vascular damage.

Multi‐Dimensional Physically Unclonable Functions: Optoelectronic Variation‐Induced Multi‐Key Generation from Small Molecule PN Heterostructures

AbstractIn this era of demanding invulnerable security systems against the threat of hacking, physically unclonable functions (PUFs), especially in which can spawn multiple security keys within a single device, has gained attention. This investigation explores the multi‐key generable PUF devices employing organic small molecules, specifically C8‐BTBT and PTCDI‐C13. The variation stems from the formation of irregular PN junctions, haphazardly configured grain boundaries of C8‐BTBT. A comprehensive analysis including scanning electron microscopy (SEM), atomic force microscopy (AFM), kelvin probe force microscopy (KPFM), impedance spectroscopy (IS), and optical simulation, has been substantiated the underlying mechanisms. Exploiting the photo‐responsive characteristics within the light wavelength spectrum of 660 and 530 nm, alongside the electrical characteristics, the capability to generate a total of 30 distinct multi‐security keys in a single device is been successfully. These keys, distinguished by variable parameters such as voltage, light wavelength, and the calculated photo‐to‐dark current ratio (PDCR), manifest novel quantitative and qualitative dimensions in security protocol customization. Inter‐Hamming distance and entropy of these cryptographic keys exhibit commendable averages of 51.9–53.1%, and 0.81, respectively. Moreover, a noteworthy average bit‐aliasing mean value of 51.9%, derived from four distinct batches, underscores the pragmatic feasibility of the proposed conceptual framework for PUF devices.