Cathepsin G, a Neutrophil Protease, Induces Compact Cell-Cell Adhesion in MCF-7 Human Breast Cancer Cells

Cathepsin G is a serine protease secreted by activated neutrophils that play a role in the inflammatory response. Because neutrophils are known to be invading leukocytes in various tumors, their products may influence the characteristics of tumor cells such as the growth state, motility, and the adhesiveness between cells or the extracellular matrix. Here, we demonstrate that cathepsin G induces cell-cell adhesion of MCF-7 human breast cancer cells resulting from the contact inhibition of cell movement on fibronectin but not on type IV collagen. Cathepsin G subsequently induced cell condensation, a very compact cell colony, resulting due to the increased strength of E-cadherin-mediated cell-cell adhesion. Cathepsin G action is protease activity-dependent and was inhibited by the presence of serine protease inhibitors. Cathepsin G promotes E-cadherin/catenin complex formation and Rap1 activation in MCF-7 cells, which reportedly regulates E-cadherin-based cell-cell junctions. Cathepsin G also promotes E-cadherin/protein kinase D1 (PKD1) complex formation, and Go6976, the selective PKD1 inhibitor, suppressed the cathepsin G-induced cell condensation. Our findings provide the first evidence that cathepsin G regulates E-cadherin function, suggesting that cathepsin G has a novel modulatory role against tumor cell-cell adhesion

Magnetization and transport properties in the superconducting Pr2_{2}Ba4_{4}Cu7_{7}O15−δ_{15-\delta} with metallic double-chain

We have reported the effect of pressure on the magnetization, and transport properties in the nominal composition Pr$_{2}$Ba$_{4}$Cu$_{7}$O$_{15-\delta}$ synthesized by a sol-gel technique. A reduction treatment of the as-sintered sample in vacuum causes higher superconductivity achieving $T_{c,on}=\sim 30$ K for $\delta =0.94$. Application of hydrostatic pressure on the oxygen depleted sample enhances its onset temperature up to 36 K at 1.2 GPa, indicating the nearly optimum doping level of the charge carrier in comparison to the pressure dependence of lower $T_{c}$ samples with $\delta =0.45$. Seebeck coefficient of the superconducting sample shows a metallic conduction, followed by a clear drop below $T_{c,on}$ and is in its temperature dependence below 100 K quite different from that of the non-superconducting one. This finding strongly suggests a dramatic change of the electronic state along the CuO double chain due to the reduction treatment for the appearance of superconductivity .Comment: 5 pages,4 figure

ALMA 0.02"-resolution observations reveal HCN-abundance-enhanced counter-rotating and outflowing dense molecular gas at the NGC 1068 nucleus

We present ALMA ~0.02"-resolution observations of the nucleus of the nearby (~14 Mpc) type-2 AGN NGC 1068 at HCN/HCO+/HNC J=3-2 lines, as well as at their 13C isotopologue and vibrationally excited lines, to scrutinize the morphological/dynamical/chemical/physical properties of dense molecular gas in the putative dusty molecular torus around a mass-accreting supermassive black hole. We confirm almost east-west-oriented dense molecular gas emission both morphologically and dynamically, which we regard as coming from the torus. Bright emission is compact (<3 pc), and low-surface-brightness emission extends out to 5-7 pc. These dense molecular gas properties are not symmetric between the eastern and western torus. The HCN J=3-2 emission is stronger than the HCO+ J=3-2 emission within the ~7 pc torus region, with an estimated dense molecular mass of (0.4-1.0)x10^6Msun. We interpret that HCN abundance is enhanced in the torus. We detect signatures of outflowing dense molecular gas and a vibrationally excited HCN J=3-2 line. Finally, we find that in the innermost (<1 pc) part of the torus, the dense molecular line rotation velocity, relative to the systemic velocity, is the opposite of that in the outer (>2 pc) part, in both the eastern and western torus. We prefer a scenario of counter-rotating dense molecular gas with innermost almost-Keplerian-rotation and outer slowly rotating (far below Keplerian) components. Our high-spatial-resolution dense molecular line data reveal that torus properties of NGC 1068 are much more complicated than the simple axi-symmetrically rotating torus picture in the classical AGN unification paradigm.Comment: 45 pages, 20 Figures. Accepted for publication in Ap

Postnatal lethality and chondrodysplasia in mice lacking both chondroitin sulfate N-acetylgalactosaminyltransferase-1 and -2

Chondroitin sulfate (CS) is a sulfated glycosaminoglycan (GAG) chain. In cartilage, CS plays important roles as the main component of the extracellular matrix (ECM), existing as side chains of the major cartilage proteoglycan, aggrecan. Six glycosyltransferases are known to coordinately synthesize the backbone structure of CS; however, their in vivo synthetic mechanism remains unknown. Previous studies have suggested that two glycosyltransferases, Csgalnact1 (t1) and Csgalnact2 (t2), are critical for initiation of CS synthesis in vitro. Indeed, t1 single knockout mice (t1 KO) exhibit slight dwarfism and a reduction in CS content in cartilage compared with wild-type (WT) mice. To reveal the synergetic roles of t1 and t2 in CS synthesis in vivo, we generated systemic single and double knockout (DKO) mice and cartilage-specific t1 and t2 double knockout (Col2-DKO) mice. DKO mice exhibited postnatal lethality, whereas t2 KO mice showed normal size and skeletal development. Col2-DKO mice survived to adulthood and showed severe dwarfism compared with t1 KO mice. Histological analysis of epiphyseal cartilage from Col2-DKO mice revealed disrupted endochondral ossification, characterized by drastic GAG reduction in the ECM. Moreover, DKO cartilage had reduced chondrocyte proliferation and an increased number of apoptotic chondrocytes compared with WT cartilage. Conversely, primary chondrocyte cultures from Col2-DKO knee cartilage had the same proliferation rate as WT chondrocytes and low GAG expression levels, indicating that the chondrocytes themselves had an intact proliferative ability. Quantitative RT-PCR analysis of E18.5 cartilage showed that the expression levels of Col2a1 and Ptch1 transcripts tended to decrease in DKO compared with those in WT mice. The CS content in DKO cartilage was decreased compared with that in t1 KO cartilage but was not completely absent. These results suggest that aberrant ECM caused by CS reduction disrupted endochondral ossification. Overall, we propose that both t1 and t2 are necessary for CS synthesis and normal chondrocyte differentiation but are not sufficient for all CS synthesis in cartilage

A Case of Urachal Carcinoma of the Abdominal Wall in a Kidney Transplant Recipient

Urachal carcinoma is an extremely rare malignant tumor arising from the urachus in the fetus. We report a patient who developed urachal carcinoma 18 years after kidney transplantation. A 59-year-old man was admitted because of abdominal pain and massive ascites. He had undergone kidney transplantation 18 years earlier and had end-stage renal disease requiring dialysis. Abdominal CT showed massive ascites and an abdominal wall cystic mass separated from the peritoneal cavity. Hemodialysis was started, and paralytic ileus was diagnosed and treated. His ileus symptoms improved temporarily, but he died of myocardial infarction. An autopsy was performed, which revealed cystadenocarcinoma in the abdominal wall mass, leading to a diagnosis of urachal carcinoma

Immunological Microenvironment Predicts the Survival of the Patients with Hepatocellular Carcinoma Treated with Anti-PD-1 Antibody

Introduction: Although immune checkpoint inhibitors (ICIs) have been considered as promising agents for the treatment of advanced hepatocellular carcinoma (HCC), previous clinical trials revealed that the response to anti-programmed cell death protein 1 (anti-PD-1) monotherapy was as low as 20%. Identifying subgroups that respond well to ICIs is clinically important. Here, we studied the prognostic factors for anti-PD-1 antibody treatment based on the molecular and immunological features of HCC. Methods: Patients who were administered anti-PD1 antibody for advanced HCC at Kindai University Hospital were included. Clinicopathological backgrounds and antitumor responses were examined in 34 cases where tumor tissues before treatment were available. Transcriptome analysis was performed using 40 HCC samples obtained from surgical resection, and immune status was compared between 20 HCCs with activating mutations in β-catenin and those without the mutations using transcriptome-based immunogram. Results: Univariate analysis showed that the disease control rate was significantly better in patients with α-fetoprotein &#x3c; 400 ng/mL, negative for β-catenin/glutamate synthetase (GS) staining, high combined positive score (CPS) of programmed death-ligand 1 (PD-L1), and increased infiltration of CD8+ cells in tumor tissues. Among them, negative staining of β-catenin/GS, CPS of PD-L1 ≥ 1, and high degree of CD8+ tumor-infiltrating lymphocytes (TILs) were significantly associated with longer survival in both progression-free survival (PFS) and overall survival (OS). The combination of these factors well stratified the survival of the patients on anti-PD-1 antibody in both PFS and OS (p &#x3c; 0.0001 and p = 0.0048 for PFS and OS, respectively). In addition, the immunogram revealed that tumor-carrying mutations in β-catenin showed downregulation of immune-related genes, especially in those related to priming and activation by dendritic cells, interferon-γ response, inhibitory molecules, and regulatory T cells. Discussion/Conclusion: The combined score including Wnt/β-catenin activation, CPS of PD-L1, and degree of CD8+ TILs in HCC is informative for predicting the response to ICI in HCC cases. Constitutive activation of β-catenin can induce an immune cold phenotype with downregulation of immune-related genes, and immunohistochemistry-based evaluation is beneficial for identifying the subgroup that shows a good response to ICI