Exopolysaccharides from Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 prevent influenza virus infection and attenuate secondary bacterial infection risk.

The present study assessed the inhibitory action of exopolysaccharides (EPS) produced by Lactobacillus delbrueckii ssp. bulgaricus OLL1073R-1 against influenza virus infection followed by secondary bacterial infection. We found that the presence of 200 or 400 µg ml-1 of EPS significantly protected against influenza virus infection in a dose-dependent manner when A549 cells were treated with EPS before infection but not after it. The expression of carcinoembryonic antigen-related cell adhesion molecule 1 (CEACAM-1), an adhesion molecule for bacteria adherence, on A549 cells was significantly enhanced during influenza virus infection compared with viral-non-infected A549 cells. However, this upregulated CEACAM-1 expression was significantly decreased by EPS treatment before viral infection in association with the reduction in the virus titre in A549 cells. In a bacterial adhesion assay using Staphylococcus aureus, the bacterial adherence to viral-infected A549 cells was significantly greater than that to viral-non-infected A549 cells, and the increased bacterial adherence induced by influenza virus infection tended to be decreased by EPS treatment before the infection. Our findings show that EPS treatment before viral infection can inhibit influenza virus infection and alleviate secondary bacterial infection through decreased CEACAM-1 expression.

Mitotic activity and specification of fibrocyte subtypes in the developing rat cochlear lateral wall.

Spiral ligament fibrocytes (SLFs) in the mammalian cochlear lateral wall participate in K(+) recycling; they are classified into five subtypes based on their morphology, distribution, and function. Regeneration of SLFs is a potential therapeutic strategy for correcting several types of hearing loss, prompting us to investigate how SLF subtypes are established during development. We compared transitional SLF-type marker expression with mitotic activity to evaluate proliferation-differentiation relationships in SLFs from postnatal rat cochleae. I.p. injection of 5-bromo-2'-deoxyuridine (BrdU) demonstrated that the overall mitotic activity of SLFs decreased significantly between postnatal day 7 (P7) and P10. For all developmental periods, BrdU incorporation was weakest in the area where type I SLFs reside. The onset of expression of markers for type II/IV SLFs followed the reduced mitotic activity of the cells, whereas that of aquaporin-1, a marker for type III SLFs, was already detectable at P7, when the type III SLFs were still proliferating vigorously. Distribution of BrdU(+) cells increased in the area of type I SLFs between P7 and P10, suggesting migration of SLFs from adjacent areas. We conclude that the time course of development of SLFs is subtype-specific.

Decreased expression of CXXC4 promotes a malignant phenotype in renal cell carcinoma by activating Wnt signaling.

The Wnt signaling pathway is involved in normal embryonic development and controls the homeostatic self-renewal of stem cells in adult tissues. Constitutive activation of Wnt signaling contributes to cancer development and progression. We identified a CXXC4 homozygous deletion at 4q24 in an aggressive renal cell carcinoma (RCC) using single-nucleotide polymorphism (SNP) arrays. CXXC4 encodes Idax, which negatively regulates Wnt signaling by binding to the PDZ domain of Dishevelled. CXXC4 mRNA levels in tumor samples were significantly lower in patients with metastases compared with those without (P=0.0016). Patients whose tumors had lower CXXC4 expression than normal kidney showed a poorer cause-specific survival outcome than those with higher expression (P=0.0095). Decreased expression of CXXC4 also correlated with cytoplasmic staining of beta-catenin. Knockdown of CXXC4 induced the nuclear translocation of beta-catenin and altered expression of a set of genes involved in cell proliferation, invasion and survival. Furthermore, reduced expression of CXXC4 by small interfering RNAs promoted cell proliferation and inhibited apoptosis after 5-FU and doxorubicin treatment in RCC cells. These data suggest that CXXC4 plays a critical role in tumor progression of RCC through Wnt signaling. Wnt signaling could thus be a potential molecular target in RCC indicating decreased CXXC4 expression.

Circulating transforming growth factor beta 1 as a predictor of liver metastasis after resection in colorectal cancer.

Plasma transforming growth factor beta1 (TGF-beta1) has been reported to be correlated with the extent of disease in colorectal cancer, but it is not known whether measuring this cytokine can help predict liver metastasis after curative resection. We prospectively studied whether plasma TGF-beta1 levels could predict liver metastasis in 117 patients with colorectal cancer before and after curative resection. Blood samples were drawn before and 2 weeks after surgery to determine the cytokine levels. Abdominal ultrasonography or computed tomography was done every 3 months after surgery. The primary end point for follow-up was recurrence. Seventy-seven of 117 cases (66%) had preoperative levels of the cytokine higher than the borderline limit of 7.5 ng/ml. Postoperative levels were >7.5 ng/ml in 29 of 117 patients (25%). The median follow-up period was 42 months (range, 5--66 months), with follow-up of all 117 patients. No recurrence was observed in 13 patients with Dukes' stage A lesions. Liver metastasis occurred in 18 of 104 patients (17%) with Dukes' stage B or C disease. Fourteen of 18 patients (78%) who developed liver metastasis had shown a postoperative plasma TGF-beta1 level of >7.5 ng/ml. Cox proportional hazards regression analysis showed that the postoperative level was a significant predictive factor for liver metastasis (P < 0.001). A single point measurement of plasma TGF-beta1 levels at 2 weeks after curative resection seems to be able to predict liver metastasis in colorectal cancer. This finding suggests the value of a prospective trial of liver-targeted adjuvant therapy for patients with elevated postoperative plasma TGF-beta1 levels.

Successful resection of a liver metastasis from gastric leiomyoblastoma: report of a case.

A 20-year-old woman was referred to our hospital for detailed investigation of a gastric submucosal tumor. A leiomyoma was preoperatively diagnosed and laparoscopic-assisted enucleation was performed. The resected tumor was 4 x 3 x 1.5 cm in size and postoperative histological examination identified it as a gastric leiomyoblastoma. Therefore, a secondary resection in the form of a distal gastrectomy was carried out. No tumor cells were found in the gastric specimen or in the lymph nodes; however, 5 months after the operation, an abdominal computed tomography scan revealed a recurrence in the liver, and she was readmitted for further examinations. The lesion was diagnosed as a single liver metastasis from the gastric leiomyoblastoma and successfully resected. The histopathological findings of the liver tumor resembled those of the primary gastric tumor. Her postoperative course was uneventful and she has been well, without any evidence of recurrence, to date. Only 12 other cases of leiomyoblastoma of the stomach with liver metastasis have been reported in Japan, all of which were associated with a very poor prognosis. Therefore, patients with this unusual disease entity should be carefully followed up after resection of the primary tumor.

Endoscopic observation of squamous metaplasia of the stomach: a report of two cases.

Squamous metaplasia of the stomach is a rare clinical entity that occurs during healing of gastric ulcers or chronic inflammation. We have treated two patients with this condition, which has only occasionally been observed endoscopically. The first was a 60-year-old woman who initially presented with multiple gastric ulcers. Two months after treatment, a white patch about 4 cm in diameter was found in the lesser curvature of the cardiac region of stomach. The second patient was a 65-year-old woman, who also developed a white patch in the same region. Two months later, a small ulcer and inflamed mucosa were seen near the lesion. The white areas in both patients were stained with Lugol's iodine solution, and biopsy specimens confirmed squamous epithelium. The squamous metaplasia was observed as a white mucosal area in the stomach, and the metaplastic area stained positively with Lugol's iodine solution.

Fecal clearance of alpha1-antitrypsin with lansoprazole can detect protein-losing gastropathy.

Detecting plasma protein loss from the stomach is quite difficult by measuring fecal alpha1-AT clearance because alpha1-AT is rapidly destroyed in the gastric juice at values below pH 3. We examined protein loss from the stomach using fecal alpha1-AT clearance with lansoprazole, a proton pump inhibitor. A 38-year-old Japanese male presented with hypoproteinemia (total serum protein: 4.4 g/dl). Abdominal scintigraphy using technetium-99m-labeled albumin revealed distinct radioactivity accumulation in the small intestine. It strongly suggested excessive protein loss to the gastrointestinal tract. Although, regular fecal alpha1-AT clearance was within the normal range (<13 ml/day), the fecal alpha1-AT clearance with the administration of lansoprazole was 80.5 ml/day. The results indicated that this method using lansoprazole is simple and useful for detecting protein-losing gastropathy.

Possible evidence for transmembrane K(+)-H+ exchange system in guinea pig myocardium.

The aim of this study was to obtain evidence for a transmembrane K(+)-H+ exchange system in Langendorff-perfused whole hearts and isolated ventricular myocytes of guinea pig. Effluent relation between K+ and pH in the whole hearts perfused with HEPES-buffered Tyrode's solution indicated a significant (p < 0.05) functional coupling of K+ uptake and H+ extrusion that was energy-dependent and omeprazole (OPZ)-sensitive. Administration of OPZ (0.3 mM) or dimethylamiloride (0.1 mM), an inhibitor of Na(+)-H+ antiport, to whole hearts subjected to the repetitive NH4Cl applications implied that both Na(+)-H+ and putative K(+)-H+ countertransports contribute to the regulation of intracellular pH. In isolated myocytes, voltage-dependent L-type Ca current (ICa) was inhibited by OPZ (0.3 mM) under K(-)- and Na(+)-free condition by 11 to 14%, and was inhibited to a greater extent (i.e., by 36 to 40%) by this agent in the presence of K+. OPZ-induced inhibition of the putative K(+)-H+ exchanger likely resulted in subsarcolemmal acidification which was responsible for the rate-independent suppression of ICa. In conclusion, these data provide functional evidence for a myocardial transmembrane K(+)-H+ exchanger.

Esophageal small cell carcinoma with ectopic production of parathyroid hormone-related protein (PTHrp), secretin, and granulocyte colony-stimulating factor (G-CSF).

A patient with primary small cell carcinoma of the esophagus is reported, in whom we have studied the secretion of a variety of hormones and cytokines. The tumor was an intermediate cell type of small cell carcinoma and had either epithelial and neuroendocrinological characteristics. Furthermore, hypercalcemia and neutrophilia were present, and the tumor was shown to produce PTHrp, secretin, and G-CSF. The present case is the first report of primary small cell carcinoma of the esophagus with ectopic production of PTHrp, secretin, and G-CSF.

Expression of vascular endothelial growth factor in human brain tumors.

Compared to normal brain an increased expression of vascular endothelial growth factor (VEGF) has been reported in many types of brain tumors. However, the numbers of samples analyzed and information about the cellular distribution of VEGF have been limited. Here we used novel monoclonal antibodies against VEGF to analyze, using immunohistochemistry, Western blotting and enzyme-linked immunosorbent assay, its expression in 108 human brain tumors that included astrocytic tumors, meningiomas, pituitary adenomas, primary intracranial germ cell tumors and neuronal tumors. The results showed that 37 of 48 astrocytic tumors (77%) and 15 of 19 meningiomas (79%) were immunoreactive for VEGF, consistent with previous reports. However, in contrast to a previous report that analyzed only VEGF mRNA; all of our 15 pituitary adenomas showed specific immunoreactivity for VEGF. We also extended the studies to previously unanalyzed neoplasms: 13 of 15 primary intracranial germ cell tumors (82%), and 7 of 10 neuronal tumors (70%) were immunoreactive for VEGF. Direct protein analysis by Western blotting confirmed the expression of VEGF in those tumors, and showed differential expression of the isoforms of VEGF protein; a pituitary adenoma expressed both VEGF165 and VEGF189 proteins, a central neurocytoma expressed only VEGF165, while an immature teratoma expressed only VEGF189. The data herein show that VEGF is expressed in a wide spectrum of brain tumors and suggest differences among tumor entities in the mechanisms of VEGF up-regulation as well as their employment of distinct VEGF isoforms for neovascularization.

Helicobacter pylori antigen in the glomeruli of patients with membranous nephropathy.

Renal biopsy specimens from patients with membranous nephropathy (MN) were studied using immunohistochemical labelling to clarify the aetiological significance of Helicobacter pylori antigen in this disease. Sixteen specimens were examined, from 7 male and 9 female MN patients. Renal specimens from patients with diabetic nephropathy and IgA nephropathy, and from autopsied patients without renal diseases were obtained as controls. Immunohistochemical labelling was performed using one polyclonal antibody and three monoclonal antibodies against H. pylori. Specimens from 11 of the MN patients revealed granular deposits along the glomerular capillary walls, which reacted positively with polyclonal antibody after trypsin pretreatment. None of the control specimens revealed positive labelling. The MN specimens showed no positive reaction with the primary antibody, which had been treated for immunoabsorption testing using sonicated H. pylori. We also determined H. pylori status in these MN patients histologically and/or serologically. Of the 11 patients whose glomeruli were positive for anti-H. pylori antibody, 7 were suitable for analysis, and all were regarded as positive for H. pylori infection. These results suggest that the presence of a specific antigen in the glomeruli of patients with MN and H. pylori infection may be involved in the pathogenesis of MN.

Coexistence of superficial esophageal carcinoma and leiomyoma: case report of an endoscopic resection.

A case of a 61 year old man who developed early esophageal cancer on top of a leiomyoma located in the upper third of the esophagus is described. Both lesions were successfully treated by endoscopic resection, without recurrence on short-term-follow-up.

Regression of duodenal mucosa-associated lymphoid tissue lymphoma after eradication of Helicobacter pylori.

It is rare for low-grade lymphoma of mucosa-associated lymphoid tissue (MALT) to affect the duodenum, and no reports have mentioned any relationship between this disease and Helicobacter pylori infection. This case report describes a patient with multiple small erosions and diffuse erythema in the duodenal bulb diagnosed histopathologically as MALT lymphoma. Immunoglobulin (Ig) heavy-chain gene rearrangement was detected, and monotypic plasma cell proliferation (IgG kappa) was shown by immunohistochemistry. The lesion was localized to the duodenal bulb. Antibiotic therapy for H. pylori resulted in resolution of the morphological features of the lymphoma, as confirmed by endoscopic and pathological examination. Moreover, the gene rearrangement could not be detected after eradication of the bacterium. Although additional follow-up is needed, it is suggested that H. pylori eradication therapy may be effective for patients with MALT lymphoma in the duodenum as well as the stomach.