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Anti-epidermal growth factor receptor (EGFR) antibody, cetuximab, therapy has significantly improved the clinical outcomes of patients with colorectal cancer, but the response to cetuximab can vary widely among individuals. We thus need strategies for predicting the response to this therapy. However, the current methods are unsatisfactory in their predictive power. Cetuximab can promote the internalization and degradation of EGFR, and its therapeutic efficacy is significantly correlated with the degree of EGFR degradation. Here, we present a new approach to predict the response to anti-EGFR therapy, cetuximab by evaluating the degree of EGFR internalization and degradation of colorectal cancer cells in vitro and in vivo. Our newly developed fluorogenic cetuximab-conjugated probe (Cetux-probe) was confirmed to undergo EGFR binding, internalization, and lysosomal degradation to yield fluorescence activation; it thus shares the action mechanism by which cetuximab exerts its anti-tumor effects. Cetux-probe-activated fluorescence could be used to gauge EGFR degradation and showed a strong linear correlation with the cytotoxicity of cetuximab in colorectal cancer cells and tumor-bearing mice. The predictive ability of Cetux-probe-activated fluorescence was much higher than those of EGFR expression or KRAS mutation status. The Cetux-probes may become useful tools for predicting the response to cetuximab therapy by assessing EGFR degradation. [ABSTRACT FROM AUTHOR]

We have developed two-photon (TP) pH-sensitive probes (BH-2 and BHEt-1) that exhibit absorption and emission maxima at 370 and 466 nm, and TP absorption cross-section values of 51 and 61 GM (1 GM = 10-50cm4s/photon), respectively, at 750 nm and pH 3.0 in a universal buffer (0.1 M citric acid, 0.1 M KH2PO4, 0.1 M Na2B4O7, 0.1 M Tris, 0.1 M KCl)/1,4-dioxane (7/3) solution. The TPM images of CCD-18co (a normal colon cell line) and HCT116 cells (a colon cancer cell line) labeled with BH-2 were too dim to be distinguished. When the same cells were labeled with BHEt-1, however, the TPM image of the HCT116 cells was much brighter than that of CCD-18co cells, and the relative proportion of the acidic vesicles (Pacid) of the former was 5-fold larger than that of latter. BHEt-1 could also differentiate HepG2 cells (a human liver cancer cell line) from LX-2 cells (a human hepatic stellate cell line) with a 6-fold larger Pacid value. Human colon cancer tissues labeled with BHEt-1 showed similar results, demonstrating much brighter TPM images and 6-fold larger Pacid values compared to normal tissue. These results suggest the potential utility of BHEt-1 for detecting colon cancer in human tissues using TPM. [ABSTRACT FROM AUTHOR]

Novel two-photon (TP) probes were developed for lysosomes (PLT-yellow) and mitochondria (BMT-blue and PMT-yellow). These probes emitted strong TP-excited fluorescence in cells at widely separated wavelength regions and displayed high organelle selectivity, good cell permeability, low cytotoxicity, and pH insensitivity. The BMT-blue and PLT-yellow probes could be utilized to detect lysosomes and mitochondria simultaneously in live tissues by using dual-color two-photon microscopy, with minimum interference from each other. [ABSTRACT FROM AUTHOR]

In this study, we developed organelle-specific blue-emitting two-photon (TP) probes for Ca2+ (BCa-1 , BCa-2 mito , and BCa-3 mem), with absorption maxima (λ max) at 350–358 nm, emission maxima (λ fl) at 464–466 nm, and TP action cross-section (Φδ max) values of 55–70 × 10−50 cm4s/photon, in the presence of excess Ca2+ at 750 nm. Moreover, the probes had dissociation constants of 0.18, 2.7, and 100 μM, respectively, which are appropriate values for sensing Ca2+ in the cytoplasm, mitochondria, and plasma membrane, respectively. The measurements were conducted using a calcium calibration buffer (10 mM 3-[ N -morpholino]propanesulfonic acid and 100 mM KCl) at pH 7.2. The TP microscopy results revealed that the probes could facilitate the real-time detection of Ca2+ in the cytoplasm, mitochondria, and plasma membranes of live cells and tissues. Additionally, we developed a green-emitting TP probe for H+ (FHEt-1 lyso) with λ max = 359 nm, λ fl = 571 nm, and Φδ max = 70 × 10−50 cm4s/photon at pH 4.3 in a universal buffer (0.1 M citric acid, 0.1 M KH 2 PO 4 , 0.1 M Na 2 B 4 O 7 , 0.1 M tris[hydroxymethyl]aminomethane, and 0.1 M KCl); this probe could detect H+ in the lysosomes. Using BCa-1 and FHEt-1 lyso , it was possible to simultaneously monitor the changes in cytosolic Ca2+ and lysosomal H+ concentrations in live cells and tissues using dual-color TP microscopy in real time. When used with TP probes emitting wavelengths of green light or longer, these blue-emitting Ca2+ probes can be used to investigate the physiological role of Ca2+ in cellular organelles as well as the crosstalk between Ca2+ and other metal ions in specific organelles. [Display omitted] • We developed organelle-specific blue-emitting two-photon Ca2+ probes. • They can detect Ca2+ in organelles of living cells and tissues in real-time. • They have appropriate K d values for Ca2+ for each organelle. • They show high sensitivity, low pH dependency, and negligible cytotoxicity. • They can monitor the crosstalk among metal ions using multicolor TPM imaging. [ABSTRACT FROM AUTHOR]

Confocal microscopy image analysis is a useful method for neoplasm diagnosis. Many ambiguous cases are difficult to distinguish with the naked eye, thus leading to high inter-observer variability and significant time investments for learning this method. We aimed to develop a deep learning-based neoplasm classification model that classifies confocal microscopy images of 10× magnified colon tissues into three classes: neoplasm, inflammation, and normal tissue. ResNet50 with data augmentation and transfer learning approaches was used to efficiently train the model with limited training data. A class activation map was generated by using global average pooling to confirm which areas had a major effect on the classification. The proposed method achieved an accuracy of 81%, which was 14.05% more accurate than three machine learning-based methods and 22.6% better than the predictions made by four endoscopists. ResNet50 with data augmentation and transfer learning can be utilized to effectively identify neoplasm, inflammation, and normal tissue in confocal microscopy images. The proposed method outperformed three machine learning-based methods and identified the area that had a major influence on the results. Inter-observer variability and the time required for learning can be reduced if the proposed model is used with confocal microscopy image analysis for diagnosis. [ABSTRACT FROM AUTHOR]

Abstract: The surface modification of Li powder was attempted in order to enhance the electrochemical properties of the electrodes used in rechargeable batteries. To create a “native” LiF film on Li powder, Li powder was manufactured using a droplet emulsion technique (DET) process by introducing fluoride as a surfactant. The surface modification of the Li powder was confirmed by means of X-ray photoelectron spectroscopy (XPS) and energy dispersive X-ray analysis (EDX). Two-electrode cells (Li symmetric cells) were prepared for the purpose of impedance analysis and scanning electron microscope (SEM) observation. From the impedance analysis, the modified surface film was found to have a smaller resistance than the bare Li powder surface. SEM images showed that dendritic formations were more highly suppressed when the compacted Li powder and surface-modified Li powder electrodes were used than when the Li foil electrode was used. Moreover, the surface-modified Li powder electrodes suppressed dendritic formations more effectively than regular Li powder electrodes. [Copyright &y& Elsevier]

The kinetics of nucleophilic substitution reactions involving 2,4‐dinitrophenyl 5‐substituted‐2‐thiophenecarboxylate were studied kinetically with 4‐Z‐C6H4O−/4‐Z‐C6H4OH, which facilitated the reactions, in 20 mol% DMSO(aq). The reactions followed second‐order kinetics and exhibited βacyl = −2.34 to −2.92, ρ(Y) = 2.71–3.39, βnuc = 0.74–0.83, and |βlg| = 0.40–0.57. Based on the interpretation of the results, we have concluded that the reaction follows an addition–elimination mechanism in which the first step is the rate‐determining step (rds). The transition state structures for the 4‐ZC6H4O−‐promoted reactions remained nearly the same with a change in the leaving group from 4‐nitrophenoxide to 2,4‐dinitrophenoxide. The mechanism of the 4‐ZC6H4O−‐promoted reaction was similar to that of R2NH‐promoted reactions, except that the former proceeded with the 1st step being the rds and the latter proceeded with a change in the rds from the second to the first step with a stronger nucleophile. [ABSTRACT FROM AUTHOR]

We report blue- and green-emitting two-photon probes derived from naphthalene and fluorene derivatives (as fluorophores) and an endoplasmic reticulum (ER) retrieval peptide (KDEL; as an ER-targeting moiety) that can detect the ER in a live cell by both one-photon and two-photon microscopy (TPM) and in a live tissue by TPM. [ABSTRACT FROM AUTHOR]

Abstract: We studied the immune-modulating effect of Maillard-type lysozyme-galactomannan conjugate (LGC). LGC significantly induced nitric oxide, and expressions of tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-8 on the murine macrophage Raw 264.7 cell line. In the mechanism of LGC, while extracellular signal-regulated kinase (ERK) was important for the induction of TNF-α, IL-1β and IL-8, the phosphorylation of C-Jun NH2-termianl kinase (JNK) contributed to the induction of TNF-α and IL-1β to a greater degree. These cytokines were less sensitive to the inhibition of p38. Nuclear factor (NF)-κB was involved in the induction of TNF-α and IL-1β. These data indicate that LGC has immune-modulating effects via JNK, ERK and NF-κB pathways, and that LGC may contribute to host immune defense. [Copyright &y& Elsevier]