Characterization of lymphocyte-rich hepatocellular carcinoma and the prognostic role of tertiary lymphoid structures.

BACKGROUND & AIMS: Lymphocyte-rich hepatocellular carcinoma (LR-HCC) is largely unknown and a rare subtype of HCC with immune-rich stroma. Tertiary lymphoid structures (TLS), frequently observed in LR-HCC, are known to be prognostically significant in various malignancies; however, their significance in HCC remains unevaluated. METHODS: Clinicopathologic data of 191 cases of surgically resected conventional HCC (C-HCC, n = 160) and LR-HCC (n = 31) were retrieved. Immunohistochemistry, multiplex immunofluorescence staining, RNA sequencing and proteomic analysis were conducted. Differences between the subtypes were statistically evaluated. RESULTS: LR-HCC was significantly correlated to larger tumour size, higher Edmondson-Steiner grade, presence of TLS and higher CD3-, CD8- and FOXP3-positive T cell, high PD-1 and PD-L1 expression (p < .001 for all) compared to C-HCC. Patients with LR-HCC exhibited significantly better overall survival (OS) (p = .044) and recurrence-free survival (RFS) (p = .025) than C-HCC. LR-HCC demonstrated TLS signatures with significantly higher proteomic-based immune scores in 14 of 17 types of tumour-infiltrating immune cells. Furthermore, C-HCC with secondary follicles, the most mature form of TLS, exhibited significantly better OS (p = .031) and RFS (p = .033) than those without. Across the global proteome, LR-HCC was well-differentiated from C-HCC and a map of protein-protein interactions between tumour-infiltrating lymphocytes and HCC in tumour microenvironment was completed. CONCLUSION: LR-HCC is clinicopathologically and molecularly distinct and shows better prognosis compared to C-HCC. Also, the presence of secondary follicle can be an important prognostic marker for better prognosis in both LR-HCC and C-HCC.

Effects of a sidestream concentrated oxygen supply system on the membrane filtration performance of a high-loaded membrane bioreactor.

To investigate the influence of high-pressure and shear effects introduced by a concentrated oxygen supply system on the membrane filtration performance, a laboratory-scale membrane bioreactor (MBR) fed artificial municipal wastewater was operated continuously for 80 days in four phases equipped with different aerations systems: (P1) bubble diffusers (days 0-40), (P2) concentrated oxygen supply system, the supersaturated dissolved oxygen (SDOX) (days 41-56), (P3) bubble diffusers (days 57-74), and (P4) SDOX (days 75-80). Various sludge physical-chemical parameters, visual inspection of the membrane, and permeability evaluations were performed. Results showed that the high-pressure effects contributed to fouling of the membranes compared to the bubble diffuser aeration system. Biofouling by microorganisms appeared to be the main contributor to the cake layer when bubble diffusers were used, while fouling by organic matter seemed to be the main contributor to the cake layer when SDOX was used. Small particle size distribution (PSD) (ranging from 1 to 10 and 1-50 µm in size) fractions are a main parameter affecting the intense fouling of membranes (e.g., formation of a dense and thin cake layer). However, PSD alone cannot explain the worsened membrane fouling tendency. Therefore, it can be assumed that a combination of several factors (which certainly includes PSD) led to the severe membrane fouling caused by the high-pressure and shear.

Tumor infiltrated immune cell types support distinct immune checkpoint inhibitor outcomes in patients with advanced non-small cell lung cancer.

The evaluation of PD-L1 expression alone has limitations in predicting clinical outcome in immune-checkpoint inhibitors (ICI). This study aimed to evaluate the predictive and prognostic effects of the presence of various immune cells in pretreatment tissue samples and to identify determinants associated with response in patients with advanced non-small cell lung cancer (NSCLC) treated with PD-1 blockade. Immune cell distribution was heterogeneous and the most dominant immune cell type was T cells. Patients with durable clinical benefit (DCB) showed significantly higher PD-L1 expression. The ratio of tumor/stroma region of T cell, B cell, and macrophage was significantly higher in patient with DCB. High intratumoral T- and B-cell density (≥median) was associated with DCB in the low PD-L1 expression (<50%) group. In univariate analyses, the overall survival (OS) benefit was shown according to intratumoral B-cell density (p = 0.0337). The incidence of hyperprogressive disease (HPD) was 13.0%. The Chi-square test revealed that HPD was significantly associated with intratumoral B-cell density but not T-cell or macrophage density. Our results demonstrate different predictive and prognostic values for infiltrating immune cells in tumor tissue, which may help in selecting patients for ICI.

Label-free imaging and evaluation of characteristic properties of asthma-derived eosinophils using optical diffraction tomography.

Optical diffraction tomography (ODT), an emerging imaging technique that does not require fluorescent staining, can measure the three-dimensional distribution of the refractive index (RI) of organelles. In this study, we used ODT to characterize the pathological characteristics of human eosinophils derived from asthma patients presenting with eosinophilia. In addition to morphological information about organelles appearing in eosinophils, including the cytoplasm, nucleus, and vacuole, we succeeded in imaging specific granules and quantifying the RI values of the granules. Interestingly, ODT analysis showed that the RI (i.e., molecular density) of granules was significantly different between eosinophils from asthma patients and healthy individuals without eosinophilia, and that vacuoles were frequently found in the cells of asthma patients. Our results suggest that the physicochemical properties of eosinophils derived from patients with asthma can be quantitatively distinguished from those of healthy individuals. The method will provide insight into efficient evaluation of the characteristics of eosinophils at the organelle level for various diseases with eosinophilia.

Double Ki-67 and synaptophysin labeling in pancreatic neuroendocrine tumor biopsies.

BACKGROUND: Pancreatic neuroendocrine tumors (PanNETs) are frequently detected on endoscopic ultrasound-guided fine-needle aspiration biopsy (EUS-FNAB) specimens. The conventional methods for evaluating the Ki-67 labeling index (Ki67LI) in EUS-FNAB specimens are laborious, and their results are difficult to interpret. More practical and easy methods for evaluating the Ki67LI in PanNETs from EUS-FNAB specimens is increasing in need. METHODS: We used double Ki-67 and synaptophysin (double Ki-Syn) antibody cocktail; Ki67LI, total Ki-67 positive cells, and total tumor cells were counted and compared with those detected on conventional single Ki-67 immunostaining (single Ki-67) of 96 PanNETs [Grade 1 (G1), 68 cases (71%); G2, 26 (27%); G3, 2 (2%)] from EUS-FNAB specimens. RESULTS: The tumor grading between double Ki-Syn and single Ki-67 immunolabeling was highly concordant (correlation, 0.95; Fisher's exact test, P < 0.001). Seven EUS-FNAB specimens (7%) had discrepant results, of which 2 were removed through surgical resection and showed the same tumor grade as that detected on double Ki-Syn immunolabeling. Fifty-four specimens (56%) had higher Ki-67 positive tumor cell counts on single Ki-67 immunolabeling. Sixty-two specimens (65%) had higher total tumor cell counts on double Ki-Syn immunolabeling. The number of specimens with less than 500 total counted tumor cells were significantly reduced when double Ki-Syn immunolabeling was applied [P = 0.046; single Ki-67, 17 specimens (18%); double Ki-Syn, 9 specimens (9%)]. CONCLUSION: Double Ki-Syn immunolabeling enables the accurate counting of the number of proliferating tumor cells without including inflammatory and contaminant epithelial cells compared with single Ki-67 immunolabeling in PanNETs from EUS-FNAB specimens.

Immune profile by multiplexed immunohistochemistry associated with recurrence after chemoradiation in rectal cancer.

BACKGROUND AND AIM: Evidence has emerged that a pretreatment immune profile in rectal cancer is associated with response to chemoradiotherapy (CRT) and recurrence after CRT. However, few studies have evaluated the immune profile differences after CRT regarding recurrence and nonrecurrence. METHODS: We included patients with advanced rectal cancer treated with CRT and surgery with recurrence within 1 year in a recurrence group. After sex and age matching with the recurrence group, patients with no recurrence for 3 years after CRT were included in a nonrecurrence group. We extracted the immune profile, including CD3 and CD8, from the surgical specimen after CRT using multispectral fluorescence immunohistochemistry and compared the two groups. RESULTS: The immune profiles of 65 patients with rectal cancer were assessed; 30 were included in the recurrence group and 35 were included in the nonrecurrence group. CD3+ and CD8+ T lymphocyte densities were significantly higher in the nonrecurrence group than in the recurrence group (CD3+ ; P < 0.001, CD8+ ; P = 0.003) in the primary tumor. Consistent results were found in epithelial and stromal cells. Compared with the recurrence group, the distinct profiles of co-expressed immune markers in the nonrecurrence group were revealed (CD3+ CD8+ , P = 0.001; CD3+ CD8+ PD-L1- , P = 0.001; CD3+ CD8+ FOXP3- PD-L1- , P = 0.001). CONCLUSIONS: Vigorous CD3+ and CD8+ T cell priming post-CRT was prominent in the nonrecurrence group compared with that of the recurrence group. This finding suggests that differences in immune profiles may have clinical significance even after CRT.

A shift from chemical oxygen demand to total organic carbon for stringent industrial wastewater regulations: Utilization of organic matter characteristics.

From 2022, industrial wastewater discharge regulations in South Korea will replace chemical oxygen demand (CODMn) with total organic carbon (TOC). A shift from CODMn to TOC is a pioneering change in protecting water bodies from organic contaminants. However, several industries are struggling to meet these TOC requirements even though their effluents met the CODMn limits. Effluent CODMn/TOC ratios (1.28 ± 0.64) found in our study were lower than the CODMn/TOC coefficients (1.33-1.80) suggested by the Ministry of Environment in South Korea. Aliphatic and particulate organic matter contents in effluents likely influenced the CODMn/TOC ratio. Regardless of the industrial category, dissolved organic carbon often consists of low molecular weight neutrals, hydrophobic organic carbon, and protein-like substances in raw and treated industrial wastewaters. The present study also revealed that TOC and CODMn represented different organic matter fractions in the paper mill and oil refinery wastewater, whereas the industrial park wastewater showed similar dissolved organic matter characteristics. Specifically, CODMn was effective in the determination of humic content in paper mill wastewater but was underestimated in oil refinery wastewater. Additionally, only paper mill effluents exceeded the TOC requirements (4 of 6 samples) and required an additional post-treatment process owing to higher organic loads.

The implications of clinical risk factors, CAR index, and compositional changes of immune cells on hyperprogressive disease in non-small cell lung cancer patients receiving immunotherapy.

BACKGROUND: Immune checkpoint blockades (ICBs) are characterized by a durable clinical response and better tolerability in patients with a variety of advanced solid tumors. However, we not infrequently encounter patients with hyperprogressive disease (HPD) exhibiting paradoxically accelerated tumor growth with poor clinical outcomes. This study aimed to investigate implications of clinical factors and immune cell composition on different tumor responses to immunotherapy in patients with non-small cell lung cancer (NSCLC). METHODS: This study evaluated 231 NSCLC patients receiving ICBs between January 2014 and May 2018. HPD was defined as a > 2-fold tumor growth kinetics ratio during ICB therapy and time-to-treatment failure of ≤2 months. We analyzed clinical data, imaging studies, periodic serologic indexes, and immune cell compositions in tumors and stromata using multiplex immunohistochemistry. RESULTS: Of 231 NSCLC patients, PR/CR and SD were observed in 50 (21.6%) and 79 (34.2%) patients, respectively and 26 (11.3%) patients met the criteria for HPD. Median overall survival in poor response groups (HPD and non-HPD PD) was extremely shorter than disease-controlled group (SD and PR/CR) (5.5 and 6.1 months vs. 16.2 and 18.3 months, respectively, P = 0.000). In multivariate analysis, HPD were significantly associated with heavy smoker (p = 0.0072), PD-L1 expression ≤1% (p = 0.0355), and number of metastatic site ≥3 (p = 0.0297). Among the serologic indexes including NLR, PLR, CAR, and LDH, only CAR had constantly significant correlations with HPD at the beginning of prior treatment and immunotherapy, and at the 1st tumor assessment. The number of CD4+ effector T cells and CD8+ cytotoxic T cells, and CD8+/PD-1+ tumor-infiltrating lymphocytes (TIL) tended to be smaller, especially in stromata of HPD group. More M2-type macrophages expressing CD14, CD68 and CD163 in the stromal area and markedly fewer CD56+ NK cells in the intratumoral area were observed in HPD group. CONCLUSIONS: Our study suggests that not only clinical factors including heavy smoker, very low PD-L1 expression, multiple metastasis, and CAR index, but also fewer CD8+/PD-1+ TIL and more M2 macrophages in the tumor microenvironment are significantly associated with the occurrence of HPD in the patients with advanced/metastatic NSCLC receiving immunotherapy.

Implication of CD69+ CD103+ tissue-resident-like CD8+ T cells as a potential immunotherapeutic target for cholangiocarcinoma.

BACKGROUND: The heterogeneous immune landscapes of intrahepatic cholangiocarcinoma (ICC) remain largely unknown. Here we aimed to investigate the implications of tissue-resident memory (TRM)-related features of tumour-infiltrating CD8+ T cells (CD8+ TILs) from ICC patients. METHODS: From ICC patients, we obtained blood samples and ICC surgical specimens (n = 33). We performed multicolour flow cytometry, multiplexed immunohistochemistry and RNA sequencing. RESULTS: When compared to peripheral CD8+ T cells, the CD8+ TILs included significantly higher proportions of the CD69+ CD103- and CD69+ CD103+ TRM-like subsets (P < .001 for both). Relative to CD69- and CD69+ CD103- cells, the CD69+ CD103+ CD8+ TILs harboured higher levels of T-cell markers representing tumour specificity (ie CD39), proliferation (ie Ki-67) and T-cell activation (ie HLA-DR and CD38) (all P < .001). Moreover, compared to the stroma, the tumour margin and core density each had a significantly higher density of CD103+ CD8+ TILs (P < .001 for both). ICCs with high proportions of CD69+ CD103+ cells displayed higher levels of parameters associated with response to immune checkpoint inhibitors (ICIs)-including number of CD8+ TIL infiltrates (P = .019), PD-L1 expression in the tumour (P = .046) and expression of the T cell-inflamed gene signature (P < .001). ICCs with lower proportions of CD69+ CD103+ CD8+ TILs exhibited significant enrichment of genes related to the Wnt/ß-catenin (P < .001) and TGF-ß pathways (P = .002). CONCLUSION: CD69+ CD103+ TRM-like CD8+ TILs represent prominent tumour-specific immune responses and hold promise as a potential therapeutic target in ICC patients. Differential TRM-related features of ICCs may help develop future immunotherapeutic strategies such as maximizing TRM responses or inhibiting pathways contributing to immune evasion.

Cyclase-associated protein 1 is a binding partner of proprotein convertase subtilisin/kexin type-9 and is required for the degradation of low-density lipoprotein receptors by proprotein convertase subtilisin/kexin type-9.

AIMS: Proprotein convertase subtilisin/kexin type-9 (PCSK9), a molecular determinant of low-density lipoprotein (LDL) receptor (LDLR) fate, has emerged as a promising therapeutic target for atherosclerotic cardiovascular diseases. However, the precise mechanism by which PCSK9 regulates the internalization and lysosomal degradation of LDLR is unknown. Recently, we identified adenylyl cyclase-associated protein 1 (CAP1) as a receptor for human resistin whose globular C-terminus is structurally similar to the C-terminal cysteine-rich domain (CRD) of PCSK9. Herein, we investigated the role of CAP1 in PCSK9-mediated lysosomal degradation of LDLR and plasma LDL cholesterol (LDL-C) levels. METHODS AND RESULTS: The direct binding between PCSK9 and CAP1 was confirmed by immunoprecipitation assay, far-western blot, biomolecular fluorescence complementation, and surface plasmon resonance assay. Fine mapping revealed that the CRD of PCSK9 binds with the Src homology 3 binding domain (SH3BD) of CAP1. Two loss-of-function polymorphisms found in human PCSK9 (S668R and G670E in CRD) were attributed to a defective interaction with CAP1. siRNA against CAP1 reduced the PCSK9-mediated degradation of LDLR in vitro. We generated CAP1 knock-out mice and found that the viable heterozygous CAP1 knock-out mice had higher protein levels of LDLR and lower LDL-C levels in the liver and plasma, respectively, than the control mice. Mechanistic analysis revealed that PCSK9-induced endocytosis and lysosomal degradation of LDLR were mediated by caveolin but not by clathrin, and they were dependent on binding between CAP1 and caveolin-1. CONCLUSION: We identified CAP1 as a new binding partner of PCSK9 and a key mediator of caveolae-dependent endocytosis and lysosomal degradation of LDLR.

Response of Transitional Mixtures Retaining Memory of In-Situ Overburden Pressure Monitored Using Electromagnetic and Piezo Crystal Sensors.

The major and minor components in granular soil materials determine their properties and behavior. This study explores the transitional behavior within threshold fines fraction of soil mixtures based on the data from the literature and experiments. From the literature survey, the void ratio, shear wave velocity, compression index, and friction angle capture the transitional turning point between the low and data-adjusted high threshold fines fractions. In particular, there is a dramatic change in hydraulic conductivity below the low threshold fines fraction that highlights the critical role of small amounts of fines in the fluid flow (e.g., clogging). From an experimental study, the engineering properties of natural soil samples identified using deformation and elastic wave sensors show transitional trends within the Revised Soil Classification System framework. The evolution of compressibility and shear wave velocity indicate that either coarse, fine, or both particles are likely to contribute to large and small strain stiffnesses when the effective stress is below 400 kPa. Thereafter, both engineering properties indicate that the soil sample retains a memory of in-situ overburden pressure when the effective stress is around 400 kPa. There is a critical role of fines that are slightly higher than low threshold fines fraction on engineering properties that promote the application of Revised Soil Classification System RSCS to natural soils.

Evaluation of Thawing and Stress Restoration Method for Artificial Frozen Sandy Soils Using Sensors.

Undisturbed frozen samples can be efficiently obtained using the artificial ground freezing method. Thereafter, the restoration of in situ conditions, such as stress and density after thawing, is critical for laboratory testing. This study aims to experimentally explore the effects of thawing and the in situ stress restoration process on the geomechanical properties of sandy soils. Specimens were prepared at a relative density of 60% and frozen at -20 °C under the vertical stress of 100 kPa. After freezing, the specimens placed in the triaxial cell underwent thawing and consolidation phases with various drainage and confining stress conditions, followed by the shear phase. The elastic wave signals and axial deformation were measured during the entire protocol; the shear strength was evaluated from the triaxial compression test. Monotonic and cyclic simple shear tests were conducted to determine the packing density effect on liquefaction resistance. The results show that axial deformation, stiffness, and strength are minimized for a specimen undergoing drained thawing, restoring the initial stress during the consolidation phase, and that denser specimens are less susceptible to liquefaction. Results highlight that the thawing and stress restoration process should be considered to prevent the overestimation of stiffness, strength, and liquefaction resistance of sandy soils.

Comparative Study on Estimation Methods of Dynamic Resistance Using Dynamic Cone Penetrometer.

Dynamic resistance, which can be used to express strength in the unit of stress and improve the reliability of the dynamic cone penetration test (DCPT), has been estimated by numerous methods. This study aims to compare different dynamic resistance estimation methods by using an instrumented dynamic cone penetrometer (IDCP). DCPTs are conducted using a standard dynamic cone penetrometer (DCP) and IDCP in the laboratory and field. Dynamic responses are obtained from the strain gauges and an accelerometer installed at the cone tip of the IDCP. The test results show that dynamic resistance is more efficient in distinguishing profiles than the dynamic cone penetration index. Among the methods to estimate the dynamic resistance at the cone tip, the force-velocity integration method and force integration method are more related to the conventional dynamic resistance considering the potential energy of the hammer than the force squared integration method. Additionally, the dynamic resistance estimated for a longer time period is more reliable, particularly for small driving rod lengths. Regarding the limitation of the dynamic response from an accelerometer in a previous study, the force-based dynamic resistance estimated for a longer time period can be used as the most reliable approach for further soil strength characterization.

Fusional Single Vision with Prism-induced Vergence Has More Influence than Diplopia on Postural Stability.

SIGNIFICANCE: Prism-induced convergence and vertical vergence for maintaining binocular fusion induced postural instability to a greater extent than diplopic conditions. This finding has important implication in relation to postural control of patients with large exophoria or vertical phoria in optometric clinic. PURPOSE: The purpose of this study was to examine whether binocular single vision with prism-induced horizontal and vertical vergence and diplopia might affect the static postural stability. METHODS: Forty-two subjects with a mean ± standard deviation age of 23.79 ± 2.81 years were enrolled in this study. To simulate convergence and divergence, base-in and base-out (BO) prisms of 4 and 8 Δ were added, respectively. To simulate vertical vergence, vertical prism of 2 Δ was added in a trial frame that corrected for the subject's refractive error during far distance (6 m) gaze. The prism power necessary to break fusion in each subject was detected and applied to induce diplopia in horizontal and vertical directions. Indices of general instability and sway power were measured under visual conditions using Tetrax and compared with normal viewing without the prism. Correlations between general instability indices and the ranges of break point in convergence and divergence were analyzed. RESULTS: Postural instability was increased significantly when convergence induced by BO 4 and 8 Δ and vertical vergence induced by 2 Δ were stimulated. The correlation coefficient between ranges of BO break point and indices of general instability was -0.308 in the BO 4 Δ and -0.306 in the BO 8 Δ condition. CONCLUSIONS: Although binocular input is recognized as an important factor in postural stability, binocular input with excessively stimulated convergence and vertical vergence during a far distance gaze is a latent factor affecting postural stability. On the other hand, diplopia did not influence postural stability.

Anti-CD45RB Antibody Therapy Attenuates Renal Ischemia-Reperfusion Injury by Inducing Regulatory B Cells.

BACKGROUND: Regulatory B cells are a newly discovered B cell subset that suppresses immune responses. Recent studies found that both anti-CD45RB and anti-Tim-1 treatments regulate immune responses by inducing regulatory B cells; however, the role of these cells in renal ischemia-reperfusion injury (IRI) is unknown. METHODS: Using mouse models, including T cell-deficient (RAG1 knockout and TCRα knockout) mice and B cell-deficient (µMT) mice, we investigated the effects of regulatory B cells and anti-CD45RB on IRI and the mechanisms underlying these effects. RESULTS: Adoptive transfer of regulatory B cells before or after IRI attenuated renal IRI. Anti-CD45RB treatment with or without anti-Tim-1 before IRI increased renal infiltration of CD19+Tim-1+ regulatory B and regulatory T cells. Anti-CD45RB decreased serum creatinine levels, pathologic injury score, tubular apoptosis, and proinflammatory cytokines levels, whereas IL-10 levels increased. Following IRI, anti-CD45RB with or without anti-Tim-1 also induced regulatory B cells, improving renal function and tubular regeneration. In RAG1 knockout mice with B cell transfer, TCRα knockout mice, and wild-type mice with T cell depletion, anti-CD45RB increased regulatory B cells and attenuated IRI. However, anti-CD45RB did not attenuate IRI in RAG1 knockout mice with T cell transfer or µMT mice and induced only mild improvement in wild-type mice with B cell depletion. Furthermore, B cell-deficient mice receiving B cells from IL-10 knockout mice (but not from wild-type mice) did not show renal protection against IRI when treated with anti-CD45RB. CONCLUSIONS: Anti-CD45RB treatment attenuated acute renal injury and facilitated renal recovery after IRI through induction of IL-10+ regulatory B cells, pointing to anti-CD45RB as a potential therapeutic strategy in renal IRI.

Variations in Velocity and Sensitivity of Electromagnetic Waves in Transmission Lines Configured in Model Piles with Necking Defects Containing Soils.

This study investigates variations in the velocity and sensitivity of electromagnetic waves in transmission lines configured in defective model piles for the detection of necking defects containing soil. Experiments are performed with model piles containing defects filled with different materials, such as air, sands, and clay. Five different types of transmission lines are configured in model piles. The electromagnetic waves are generated and detected using a time domain reflectometer. The velocity of electromagnetic waves is highest when the defect is filled with air, and it decreases with an increase in the water content. The velocity is lowest when the defect is filled with clay. The sensitivity of transmission lines for detecting defects decreases with an increase in soil water contents. The transmission line with a single electrical wire and epoxy-coated rebar exhibits the highest sensitivity, followed by that with three and two parallel electrical wires. Transmission lines with a single electrical wire and uncoated rebar and those with two parallel electrical wires wrapped with a sheath exhibit poor sensitivity when the defect is filled with clay. This study demonstrates that electromagnetic waves can be effective tools for detecting necking defects with wet and conductive soils in bored piles.

Strength Characteristics of Sand-Silt Mixtures Subjected to Cyclic Freezing-Thawing-Repetitive Loading.

Daily freezing-thawing-repetitive loading is a critical factor affecting soil stability. This study assesses the strength of sand-silt mixtures with various silt fractions (SFs) subjected to cyclic freezing-thawing-repetitive loading. Specimens with SF of 0-100% were prepared with a fixed relative density of 60%. The number of repetitive loadings (N) was 1, 100, and 1000 for each specimen with different SFs. After three cycles of freezing-thawing-repetitive loading, the specimens were frozen at -5 °C for the uniaxial compression test. Test results show that the change in relative density (∆Dr) increases with the increase in SF up to 30% and decreases as SF increases beyond 30% owing to the change in the void ratio. The volumetric unfrozen water content (θu) increases with the increase in both SF and N owing to the effect of the physicochemical characteristics of soils on small voids. Unconfined compressive strength of sand-dominant mixtures (SF ≤ 30%) is reinforced by ∆Dr. By contrast, for silt-dominant mixtures (SF > 30%), the unconfined compressive strength decreases with the increase in θu and N due to lubricant role and sands dispersion. Thus, the effects of SF and N should be considered for sand-silt mixtures that have a probability to undergo cyclic freezing-thawing-repetitive loading.

Advances in Analysis of Biodistribution of Exosomes by Molecular Imaging.

Exosomes are nano-sized membranous vesicles produced by nearly all types of cells. Since exosome-like vesicles are produced in an evolutionarily conserved manner for information and function transfer from the originating cells to recipient cells, an increasing number of studies have focused on their application as therapeutic agents, drug delivery vehicles, and diagnostic targets. Analysis of the in vivo distribution of exosomes is a prerequisite for the development of exosome-based therapeutics and drug delivery vehicles with accurate prediction of therapeutic dose and potential side effects. Various attempts to evaluate the biodistribution of exosomes obtained from different sources have been reported. In this review, we examined the current trends and the advantages and disadvantages of the methods used to determine the biodistribution of exosomes by molecular imaging. We also reviewed 29 publications to compare the methods employed to isolate, analyze, and label exosomes as well as to determine the biodistribution of labeled exosomes.

Increased nicotinamide adenine dinucleotide pool promotes colon cancer progression by suppressing reactive oxygen species level.

Nicotinamide adenine dinucleotide (NAD) exists in an oxidized form (NAD+ ) and a reduced form (NADH). NAD+ plays crucial roles in cancer metabolism, including in cellular signaling, energy production and redox regulation. However, it remains unclear whether NAD(H) pool size (NAD+ and NADH) could be used as biomarker for colon cancer progression. Here, we showed that the NAD(H) pool size and NAD+ /NADH ratio both increased during colorectal cancer (CRC) progression due to activation of the NAD+ salvage pathway mediated by nicotinamide phosphoribosyltransferase (NAMPT). The NAMPT expression was upregulated in adenoma and adenocarcinoma tissues from CRC patients. The NADH fluorescence intensity measured by two-photon excitation fluorescence (TPEF) microscopy was consistently increased in CRC cell lines, azoxymethane/dextran sodium sulfate (AOM/DSS)-induced CRC tissues and tumor tissues from CRC patients. The increases in the NAD(H) pool inhibited the accumulation of excessive reactive oxygen species (ROS) levels and FK866, a specific inhibitor of NAMPT, treatment decreased the CRC nodule size by increasing ROS levels in AOM/DSS mice. Collectively, our results suggest that NAMPT-mediated upregulation of the NAD(H) pool protects cancer cells against detrimental oxidative stress and that detecting NADH fluorescence by TPEF microscopy could be a potential method for monitoring CRC progression.

Association between the exposure to anti-angiogenic agents and tumour immune microenvironment in advanced gastrointestinal stromal tumours.

BACKGROUND: Tumour immune microenvironment (TIME) of gastrointestinal stromal tumours (GISTs) is largely unknown. METHODS: A total of 81 surgical specimens from 67 patients with advanced GISTs were categorised into treatment groups: tyrosine kinase inhibitor (TKI)-naive, n = 20; imatinib-progressed and no exposure to sunitinib or regorafenib (IM-PD), n = 30; and imatinib-progressed and sunitinib and/or regorafenib-treated (IM-PD/SU-treated), n = 31. Multiplexed immunofluorescence staining and RNA sequencing were performed to define TIME. RESULTS: PD-L1 expression rate (>1%) of DOG-1+ tumour cells was 5.0, 6.7, and 29.0% in TKI-naive, IM-PD, and IM-PD/SU-treated group, respectively (p = 0.02). FoxP3 expression of CD3+ T cells and CD204+ CD68+ monocytes per DOG-1+ cells was significantly higher in IM-PD/SU-treated group compared to TKI-naive and IM-PD groups (p < 0.05). IM-PD/SU-treated group showed increased expression of PD-1 on CD3+ T cells (p = 0.03 vs TKI-naive; p = 0.003 vs IM-PD) and DOG-1+ tumour cells (p = 0.02 vs TKI-naive; p = 0.006 vs IM-PD), TIM-3 expression on CD3+ T cells (p = 0.01 vs TKI-naive; p = 0.002 vs IM-PD), and LAG3 expression on CD3+ T cells (p = 0.001 vs TKI-naive; p = 0.004 vs IM-PD). In the RNAseq analysis, TIGIT expression was significantly increased in IM-PD/SU-treated GISTs compared to IM-PD (p = 0.01). CONCLUSION: Immunosuppressive phenotype was predominant in tumours treated with anti-angiogenic agents compared to TKI-naive and IM-treated tumours.