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Capillary electrophoresis using laser-induced fluorescence detection (CE-LIF) is one of the most sensitive separation tools among electrical separation methods. The use of CE-LIF in immunoassays and enzyme assays has gained a reputation in recent years for its high detection sensitivity, short analysis time, and accurate quantification. Immunoassays are bioassay platforms that rely on binding reactions between an antigen (analyte) and a specific antibody. Enzyme assays measure enzymatic activity through quantitative analysis of substrates and products by the reaction of enzymes in purified enzyme or cell systems. These two category analyses play an important role in the context of biopharmaceutical analysis, clinical therapy, drug discovery, and diagnosis analysis. This review discusses the expanding portfolio of immune and enzyme assays using CE-LIF and focuses on the advantages and disadvantages of these methods over the ten years of existing technology since 2008.

Mesenchymal stem cells (MSCs) have been developed as cell therapeutics for various immune disorders using their immunoregulatory properties mainly exerted by their paracrine functions. However, variation among cells from different donors, as well as rapid clearance after transplantation have impaired the uniform efficacy of MSCs and limited their application. Recently, several strategies to overcome this limitation have been suggested and proven in pre-clinical settings. Therefore, in this review article, we will update the knowledge on bioengineering strategies to improve the immunomodulatory functions of MSCs, including genetic modification and physical engineering.

Pancreatic cancer remains a highly lethal malignancy due to late diagnosis and the lack of effective non-invasive biomarkers for detection, further complicated by biological heterogeneity, including intraductal papillary mucinous neoplasm (IPMN) and IPMN-associated invasive carcinoma (IPMC). Serum samples from a discovery cohort (n = 60), including patients with IPMN, IPMC, and pancreatic ductal adenocarcinoma (PDAC), were analyzed using LC-MS/MS-based serum proteomic profiling to identify candidate biomarkers. Feature selection was performed using ANOVA, receiver operating characteristic analysis, and permutation feature importance. The selected markers were further evaluated in an independent validation cohort (n = 40). In the discovery cohort, the model demonstrated robust performance, with leave-one-out cross-validation AUROC values ranging from 0.814 to 1.000 across classifications of healthy controls and disease groups (IPMN, IPMC, and PDAC). In the independent validation cohort, the six-marker panel (CALR, FCN1, MBL2, SPP1, TAGLN2, and VWF) achieved an AUROC of 0.962 with a specificity of 95.0% for distinguishing healthy controls from IPMC, and an AUROC of 0.856 with a specificity of 90.0% for healthy controls vs. PDAC. These findings demonstrate the robustness and clinical potential of the six-protein panel as a non-invasive diagnostic tool for pancreatic cancer.

This study evaluated the effects of the light intensity of curing and the post-curing duration on the mechanical properties and accuracy of the interim dental material. After designing the specimen, 3D printing was performed, and the light intensity was divided into groups G20, G60, G80, and G120 (corresponding to 1.4–1.6, 2.2–3.0, 3.8–4.4, and 6.4–7.0 mW/cm2, respectively), with no post-curing or 5, 10, or 20 min of post-curing being performed. The flexural properties, Vickers microhardness, degree of conversion (DC), and 3D accuracy were then evaluated. The flexural properties and Vickers microhardness showed a sharp increase at the beginning of the post-curing and then tended to increase gradually as the light intensity and post-curing time increased (p < 0.001). On the other hand, there was no significant difference between groups in the accuracy analysis of a 3D-printed three-unit bridge. These results indicate that the light intensity of the post-curing equipment influences the final mechanical properties of 3D-printed resin and that post-curing can be made more efficient by optimizing the light intensity and post-curing time.

Colorectal cancer (CRC) is a major global cause of death, with metastases and chemotherapy resistance contributing to poor outcomes. To identify natural compounds with anticancer potential against CRC and elucidate their action mechanisms, the cytotoxicity of 37 natural compounds was evaluated against the HCT116, leading to the identification of conferone as the lead candidate. Its anti‐migratory and anti‐invasive effects were evaluated in HCT116, Colo205, and SW480 cells. The interactions between conferone and focal adhesion kinase (FAK) were assessed through protein expression analysis and molecular docking. Glutaminolysis regulation was determined by LC–MS/MS, and related enzyme levels were detected by western blotting. Conferone inhibited migration and invasion in all three CRC cell lines, though it showed limited anti‐proliferative activity. At 10 μM, conferone reduced FAK and p‐FAK (Tyr397) protein levels, reversing the epithelial‐mesenchymal transition. Docking analysis confirmed direct FAK binding and predicted inhibition of its phosphorylation, with greater affinity than the FAK inhibitor 1,2,4,5‐benzene tetramine tetrahydrochloride. Conferone also downregulated glutaminase and glutamate–ammonia ligase, increasing glutamine and decreasing glutamic acid. Additionally, it suppressed c‐raf phosphorylation and reduced c‐Myc expression, blocking glutaminolysis‐driven metabolism. These findings highlight conferone as a potential therapeutic agent that targets FAK, alters metabolic reprogramming, and impedes CRC progression.

This study investigates the effects of different temperature conditions on the quality of black garlic (BG) during the aging process. Two temperature protocols were employed: gradual heating and cooling (GHC), where the temperature was slowly raised from 45 °C to 77 °C and then lowered to 59 °C at a rate of 1 °C per hour, and rapid heating and cooling (RHC), where the temperature was quickly raised from 45 °C to 85 °C and then lowered to 56 °C at a rate of 1 °C every 30 min. Changes in surface color, hardness, moisture, pH, fructose, total polyphenol content (TPC), and key sulfur compounds such as alliin, S-allylcysteine (SAC), and γ-glutamyl-S-allylcysteine (γ-GSAC) were analyzed. Our findings showed that GHC led to a higher increase in TPC and fructose content by the 15th day compared to RHC. In contrast, RHC retained significantly higher SAC concentrations, approximately 1.7 times that of GHC, by the end of the aging period. Surface color changes, particularly in lightness and redness, were more pronounced under GHC, while RHC demonstrated superior moisture retention. These findings indicate that GHC is better suited for products prioritizing polyphenols and sugars, while RHC is more optimal for SAC-enriched BG. This study provides valuable insights into optimizing BG production for diverse food and medicinal applications through precise temperature modulation.

The role of matrix metalloproteinase-2 (MMP-2) in tumor cell migration has been widely studied, however, the characteristics and effects of MMP-2 in clinical sample of metastatic colorectal cancer (CRC) remain poorly understood. Here, in order to unveil the perturbed proteomic signal during MMP-2 induced cancer progression, we analyzed plasma proteome of CRC patients according to disease progression, HCT116 cancer secretome upon MMP-2 knockdown, and publicly available CRC tissue proteome data. Collectively, the integrative analysis of multi-layered proteomes revealed that a protein cluster containing EMT (Epithelial-to-Mesenchymal Transition)-associated proteins such as CD9-integrin as well as MMP-2. The proteins of the cluster were regulated by MMP-2 perturbation and exhibited significantly increased expressions in tissue and plasma as disease progressed from TNM (Tumor, Node, and Metastasis) stage I to II. Furthermore, we also identified a plausible association between MMP-2 up-regulation and activation of focal adhesion kinase signaling in the proteogenomic analysis of CRC patient tissues. Based on these comparative and integrative analyses, we suggest that the high invasiveness in the metastatic CRC resulted from increased secretion of MMP-2 and CD9-integrin complex mediated by FAK signaling activation.

Glioblastoma (GBM) is the most common and the deadliest brain cancer. Glutamine anabolism mediated by glutamine synthetase (GS) is beneficial for GBM cell growth, especially under glutamine deprivation. However, the molecular mechanism underlying GS homeostasis in GBM remains undisclosed. Here, it is reported that histone deacetylase 6 (HDAC6) promotes GS deacetylation, stabilizing it via ubiquitin‐mediated pathway. It is found that deubiquitination of GS is modulated by ubiquitin‐specific peptidase 9, X‐linked (USP9X). USP9X stabilizes GS by removing its K48‐linked polyubiquitination on lysine 91 and 103. Accordingly, targeting HDAC6 and USP9X in vitro and in vivo represses GBM tumorigenesis by decreasing GS stability. Metabolic analysis shows that silencing HDAC6 and USP9X disrupts de novo nucleotide synthesis, thereby attenuating GBM cell growth. Furthermore, GS modulation by targeting HDAC6 and USP9X restrains the self‐renewal capacity. These results suggest that HDAC6 and USP9X are crucial epigenetic enzymes that promote GBM tumorigenesis by modulating glutamine metabolism.

We developed the microfluidic co-culture platform to study photothermal therapy applications. We conjugated folic acid (FA) to target breast cancer cells using reduced graphene oxide (rGO)-based functional nanomaterials. To characterize the structure of rGO-based nanomaterials, we analyzed the molecular spectrum using UV–visible and Fourier-transform infrared spectroscopy (FT-IR). We demonstrated the effect of rGO-FA-based nanomaterials on photothermal therapy of breast cancer cells in the microfluidic co-culture platform. From the microfluidic co-culture platform with breast cancer cells and human umbilical vein endothelial cells (HUVECs), we observed that the viability of breast cancer cells treated with rGO-FA-based functional nanomaterials was significantly decreased after near-infrared (NIR) laser irradiation. Therefore, this microfluidic co-culture platform could be a potentially powerful tool for studying cancer cell targeting and photothermal therapy.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) nucleocapsid protein (NP) participates in viral genome packaging and abundantly produced when infected. In this work, SPR biosensor for the detection of SARS-CoV-2 in viral fluid using Fv-antibodies with the binding affinity to nucleocapsid protein (NP) of SARS-CoV-2. The F V -antibodies with a specific binding activity to the SARS-CoV-2 NP were screened using the F V -antibody library, which was expressed on the outer membrane of E. coli . F V -antibodies comprised three complementarity-determining regions (CDRs) and four frame regions (FRs) of the heavy chain at the binding pocket of IgG. The F V -antibody library was prepared by performing site-directed mutagenesis and by using the autodisplay technology; F V -antibodies with specific binding activities to the nucleocapsid protein (NP) of SARS-CoV-2 were screened using NP-immobilized magnetic beads. First, E. coli isolates with the target F V -antibody were screened, and the binding affinity (K D ) was estimated for the screened E. coli clones using FACS analysis. Then, the outer membrane (OM) of the screened E. coli clones with autodisplayed Fv-antibodies was obtained and layered on an SPR biosensor, and the binding curves of four different coronavirus (CoV) culture fluids, SARS-CoV-2, SARS-CoV, MERS-CoV, and CoV strain 229E, were compared. Finally, the F V -antibodies of the screened E. coli clones were synthesized as peptides (11 amino acid residues), and the binding constants (K D ) to NP as well as the binding curves of the CoV strains in culture fluids were estimated. Using docking simulation, binding sites and interaction types between NP and each synthetic peptide were investigated. Graphical Abstract