BACKGROUND & AIMS:The incidence of gastric cardia adenocarcinoma (GCA) and metabolic syndrome (MetS) have increased concurrently. Lipopolysaccharide (LPS) may be a biomarker for GCA development and treatment resistance of immune checkpoint inhibitors. We aimed to elucidate whether high-fat diet (HFD)-related endotoxemia progresses GCA via programmed death-ligand 1 (PD-L1) signaling. METHODS:We studied K19-Wnt1/C2mE mice fed HFD or control diet with or without systemic administration of Escherichia coli LPS ± clodronate liposomes (CLs). To clarify the role of nuclear factor E2-related factor 2 (Nrf2) in tumorigenesis, we generated Nrf2-deficient K19-Wnt1/C2mE mice. We performed further experiments using MKN7 and THP-1 cells. RESULTS:In the HFD + LPS group of K19-Wnt1/C2mE mice, tumor growth with increase in tumor cell proliferation and macrophage infiltration was observed with gut dysbiosis, gut mucosal barrier damage, endotoxemia, and insulin resistance. The expression of 8-OHdG, NAD(P)H quinone oxidoreductase 1 (Nqo1), Tumor necrosis factor (TNF)-a, phosphorylated nuclear factor-kappa B (p-NFκB), and PD-L1 was upregulated in tumors. TNF-α expression was observed in tumor cells and stromal macrophages. These effects were abolished in the HFD + LPS + CL group. In the Nrf2-deficient K19-Wnt1/C2mE mice, tumors shrank as TNF-α and PD-L1 expression decreased, without improvement of gut barrier damage and endotoxemia. LPS stimulation of phorbol 12-myristate 13-acetate-treated THP-1 cells induced the expression of TNF-α, which activated NFκB-PD-L1 signaling in cocultured MKN7 cells. LPS stimulation of MKN7 cells increased the expression of NQO1, phosphorylated NFκB, and PD-L1, and NRF2 directly regulated CD274 transcription. CONCLUSIONS:HFD-related metabolic endotoxemia may promote GCA progression via PD-L1 induction in tumor cells directly through Nrf2 signaling activated by LPS and through NFκB signaling by TNF-α from LPS-activated macrophages in tumor microenvironment.

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Supplementary Data from Age-Related EBV-Associated B-Cell Lymphoproliferative Disorders Constitute a Distinct Clinicopathologic Group: A Study of 96 Patients

Atypical lipomatous tumor/well-differentiated liposarcoma (ALT/WDLPS) is usually a solitary adipocytic tumor. ALT/WDLPS shows no potential for metastasis unless it undergoes dedifferentiation. No case of multiple ALT/WDLPS has been reported in recent years. We present a rare case of multiple recurrent liposarcomas. A 71-year-old man with a history of scrotal ALT/WDLPS at 61 years presented with multiple large tumors spread throughout the body. The patient was bedridden and severely limited in his activities of daily living (ADL) due to multiple large tumors in the trunk and lower extremities. Radiological examination revealed multiple adipocytic tumors, mainly in the soft tissues of the trunk and extremities, with several visceral lesions. Tumors were resected in stages, starting with large tumors directly related to disability. Repeated palliative resections improved the patient’s ADL; he regained ambulation and was discharged 18 months after admission. Twelve surgeries were performed to remove 44 adipocytic tumors from the testis, left chest wall, perigastric area, ileum, left inguinal region, both buttocks, thighs, and lower legs. Histological examination revealed dedifferentiated components in five tumors, while 39 tumors were diagnosed as ALT/WDLPS. At the age of 76 years, the patient developed an unresectable dedifferentiated liposarcoma between the heart and aorta, leading to fatality at 79 years. The patient’s clinical course suggested multiple metastases of ALT/WDLPS of scrotal origin or ALT/WDLPS of multicentric origin. Although multicentric ALT/WDLPS or ALT/WDLPS metastases are rare, they should be considered when multiple large adipocytic tumors are found throughout the body. Despite the presence of numerous large malignant tumors, surgical treatments of the lesions can improve ADL and prolong life if the tumors are of low-grade malignancy.

MUM1/IRF4 expression is detected in 18% to 41% of Burkitt lymphoma (BL). However, only a few studies of MUM1-positive (MUM1 + ) BL have been reported, and its characteristics still remain controversial. To highlight the features of MUM1 + BL, we compared the clinicopathologic characteristics of 37 cases of MUM1 + and 51 cases of MUM1-negative (MUM1 − ) BL in Japan. Compared with MUM1 − BL, patients with MUM1 + BL showed significantly younger onset ( P =0.0062) and a higher ratio of females ( P =0.013). We have also revealed the difference in the involved sites. The MUM1 + group showed lower incidences of involvement of stomach ( P =0.012) and tonsil ( P =0.069). There was a more tendency in MUM1 + group to involve colon ( P =0.072), breast ( P =0.073), and kidney ( P =0.073). Regarding the prognosis, a trend toward a lower overall survival for MUM1 + group was noted ( P =0.089). Notably, comparing MUM1 + and MUM1 − BL cases of adults (age16 y old and above), the former showed significantly worse prognosis ( P =0.041). Among the BL patients treated with the intensive chemotherapy, a standard therapy for BL, MUM1 + cases showed worse prognosis ( P =0.056). In conclusion, MUM1 + BL showed worse prognosis, particularly in adult cases, compared with MUM1 − BL. In addition, the difference of the onset age, sex ratio, and involved sites between the 2 groups was noted. Our results demonstrate that MUM1 expression might predict worse prognosis of BL, and MUM1 + BL should be distinguished from MUM1 − BL.

It is well established that polymorphisms of the caspase activation and recruitment domain 15 (CARD15) gene, a major risk factor in Crohn's disease (CD), lead to loss of nucleotide-binding oligomerization domain 2 (NOD2) function. However, a molecular explanation of how such loss of function leads to increased susceptibility to CD has remained unclear. In a previous study exploring this question, we reported that activation of NOD2 in human dendritic cells by its ligand, muramyl dipeptide (MDP), negatively regulates Toll-like receptor (TLR)-mediated inflammatory responses. Here we show that NOD2 activation results in increased interferon regulatory factor 4 (IRF4) expression and binding to tumor necrosis factor receptor associated factor 6 (TRAF6) and RICK (receptor interacting serine–threonine kinase). We then show that such binding leads to IRF4-mediated inhibition of Lys63-linked polyubiquitination of TRAF6 and RICK and thus to downregulation of nuclear factor (NF)-κB activation. Finally, we demonstrate that protection of mice from the development of experimental colitis by MDP or IRF4 administration is accompanied by similar IRF4-mediated effects on polyubiquitination of TRAF6 and RICK in colonic lamina propria mononuclear cells. These findings thus define a mechanism of NOD2-mediated regulation of innate immune responses to intestinal microflora that could explain the relation of CARD15 polymorphisms and resultant NOD2 dysfunction to CD.

Oxidative stress might participate in the carcinogenesis of human esophageal squamous cell carcinomas (hESCC). 4‐Hydroxynonenal (HNE) is a major product of membrane lipid peroxidation with short life. It might act as an important mediator through the generation of adducts and activate epidermal growth factor receptor (EGFR) signaling. It is mainly trapped with glutathione (GSH) and catalyzed by glutathione S ‐transferases (GSTs). This study aimed to elucidate the possible participation of HNE, GSH/GST system, and EGFR signaling in hESCC development. Immunohistochemistry of HNE adducts, EGFR, and phosphorylated EGFR (pEGFR) was performed with hESCC specimens. The effect of HNE on the phosphorylation of EGFR and its downstream PhospholipaseCγ1 (PLCγ1) was investigated with KYSE30 cell‐line. Pretreatment with GSH inducer N ‐acetylcysteine (NAC) or GSH inhibitor Buthionine sulfoximine (BSO) and mandatory transfection of hGSTA4 gene in KYSE30 were conducted to investigate the relationship between HNE and GSH/GST system. Immunoreactants of HNE adducts, EGFR, and pEGFR were increased in hESCC compared to non‐cancerous epithelium with positive correlations. The treatment of HNE ligand‐independently induced the phosphorylation of EGFR and PLCγ1 accompanying the diminishment of intracellular GSH level. NAC increased GSH contents but BSO decreased in dose‐dependent manners. Reflecting changes in GSH, HNE‐induced EGFR phosphorylation was suppressed by NAC, whereas it was promoted by BSO. Mandatory expression of hGSTA4 suppressed HNE‐induced events. We first demonstrated that the ligand‐independent activation of EGFR by the balance between the stimulation of HNE and the prevention of intrinsic GSH/GST system might participate in the development of hESCC. © 2011 Wiley‐Liss, Inc.

To report the results of radiation therapy with concurrent intermittent arterial chemotherapy using a reservoir for muscle-invasive bladder cancer. From Apr. 1997 to Sep. 2006, we analyzed the clinical results of 11 patients (8 males and 3 females), ranged in age 61∼83 (mean 78 years,). All 11 patients had urothelial cancer. The TNM classifications were with T stage; T1b: 1, T2a: 5, T2b: 2, T3a: 1, T4a: 2. All were classified N0 M0. Pathologic grades were G2 in 5 patients and G3 in 6. The nonsurgical reason was due to advanced age in 5 patients, refusal of surgery in 5 and multiple primary cancers in 1. In prior to treatment, arterial blood supply was altered by coil embolizations to one vessel for bladder. The contralateral internal iliac artery and the ipsilateral superior gluteal artery were embolized by coil, and the tip of catheter was fixed inside the superior gluteal artery and side hole of the catheter was indwelled into the internal iliac artery. Then, a reservoir was placed in a lower abdominal wall subcutaneous pocket. Irradiation was limited to the bladder using four opposed field techniques with 2 Gy x 25 = 50 Gy. In the meantime, 10mg/body ia THP on the days 2,9,16 and 23 and 20mg/body ia CDDP on the days 3, 10, 17, 24 were administered concurrently through the reservoir. The follow-up period ranged from 17 to 117 months. All patients achieved a CR due to the initial treatment. Only 2 of 11 patients had local recurrence: one treated with TUR-Bt, and the other became invasive after TUR-Bt and underwent total cystectomy. Consequently, 2 of 2 patients who had local recurrence were salvaged. Although only minor acute side-effects (microhematuria) was found, this was thought to be the late side-effects due to the deterioration of the contracted bladder caused by the precedent TUR-Bt. The following two complications caused by the intra-arterial chemotherapy were found: the reservoir was removed due to infection after treatment in one patient. In another, because of the positive cytology one month after treatment, COMPA chemotherapy was intended to add optional treatment, but perianal burning sensation and skin ulcer were appeared, probably caused by the recanalization of the embolized vessel. This resulted in the halt of treatment, but obtained a CR. Distant metastasis was found in three patients. Disease-specific death was seen in two patients, intercurrent-disease death in two, disease-free survivor in six and being alive with cancer in one. These findings suggest that radiation therapy with concurrent arterial chemotherapy for invasive bladder cancer produces an excellent local effect with less side effects. This therapy could be an alternative to bladder-preservation treatment.

Nucleotide oligomerization domain (NOD)2 is a member of the NOD-like receptor family of proteins that initiate inflammatory responses when exposed to ligands derived from bacterial components that gain access to the intracellular milieu. It is thus somewhat paradoxical that polymorphisms in the gene that encode NOD2 (CARD15) that lead to impaired NOD2 function, are susceptibility factors in Crohn's disease, a condition marked by excessive inflammatory responses to normal bacterial flora. In an initial series of studies conducted in our laboratory to better define NOD2 function and to resolve this paradox we showed that NOD2 activation by its ligand, muramyl dipeptide (MDP) ordinarily downregulates responses to Toll-like receptor (TLR) stimulation, and thus cells lacking NOD2 mount increased responses to such stimulation. This fits with the fact that mice bearing an NOD2 transgene, and thus having cells with increased NOD2 function display decreased responses to TLR stimulation and are resistant to experimental colitis induction. In further studies, we showed that prestimulation of cells with NOD2 ligand renders them unresponsive to TLR stimulation, because such prestimulation results in the elaboration of inhibitory factor (IRF4), an inhibitor of TLR-induced inflammatory pathways. Furthermore, administration of MDP to normal mice induces IRF4 and prevents experimental colitis. These studies strongly suggest that NOD2 polymorphisms are associated with Crohn's disease because they lead to a decrease in the negative regulation of TLR responses occurring in the normal gut, and thus a pathologic increase in responses to the normal flora. The finding that MDP administration prevents experimental colitis opens the door to the possibility that such treatment might quell Crohn's disease relapses in patients without NOD2 abnormalities.

Micronucleus induction was studied for the DNA target clastogens mitomycin C (MMC) and 1-beta-D-arabinofuranosylcytosine (Ara-C), and also the non-DNA target aneugen colchicine (COL) in order to evaluate the dose-response relationship at very low dose levels. The acridine orange (AO) supravital staining method was used for microscopy and the anti-CD71-FITC based method was used for flow cytometric analysis. In the AO method, 2000 reticulocytes were analysed as commonly advised, but in the flow cytometric method, 2000, 20 000, 200 000 and 1 000 000 reticulocytes were analysed for each sample to increase the detecting power (i.e. sensitivity) of the assay. The present data show that increasing the number of cells scored increases the statistical power of the assay when the cell was considered as a statistical unit. Even so, statistically significant differences from respective vehicle controls were not observed at the lowest dose level for MMC and Ara-C, or the lower four dose levels for COL, even after one million cells were analysed. When the animal was considered as a statistical unit, only the top dose group for each chemical showed significant increase of micronucleated reticulocytes frequency. As non-linear dose-response curves were obtained for each of the three chemicals studied, these observations provide evidence for the existence of a practical threshold for the DNA target clastogens as well as the non-DNA target aneugen studied.

We investigated the effectiveness of fosfomycin combined with other antibiotics, such as piperacillin, cefepime, ceftazidime, imipenem, meropenem, aztreonam, gentamicin, or levofloxacin, against 30 Pseudomonas aeruginosa strains, including multidrug-resistant strains, isolated from clinical specimens, using the efficacy time index (ETI) assay. The assay refers to the result of pharmacokinetics obtained from adult men volunteers, and yields an ETI to evaluate the effect of a combination of antimicrobial agents. With the ETI, based on serum concentration 3 h after the administration of two antimicrobial agents, the effectiveness of antimicrobial combinations was evaluated as follows: poor, ETI < 0.5; fair, 0.5 ≤ ETI < 1; good, 1 ≤ ETI < 8; and excellent, ETI ≥ 8. The combination of fosfomycin and cefepime (efficacy rate [excellent plus good], 76.7%) and fosfomycin/aztreonam (efficacy rate, 76.7%) appeared to be the most effective, followed by fosfomycin/meropenem (efficacy rate, 76.6%), fosfomycin/imipenem (efficacy rate, 73.3%), fosfomycin/ceftazidime (efficacy rate, 70%), fosfomycin/gentamicin (efficacy rate, 70%), fosfomycin/piperacillin (efficacy rate, 66.7%), and fosfomycin/levofloxacin (efficacy rate, 66.7%). Fosfomycin/cefepime, fosfomycin/aztreonam, and fosfomycin/meropenem may be clinically useful in selected patients, particularly for P. aeruginosa. The ETI assay provided information on the minimum inhibitory concentration (MIC) of many pairs of combined antimicrobial agents simultaneously. The ETI assay may be a useful technique with which to investigate the effect of combinations of antimicrobial agents against P. aeruginosa, including multidrug-resistant strains.