The chemiluminescence of the Cypridina luciferin analogue, 2-methyl-6-(p-methoxyphenyl)-3,7-dihydroimidazo[1,2-a]pyrazin++ +-3-one (MCLA) was observed at 462 nm in the presence of horseradish peroxidase (HRP) and the total spectrum of light emitted was found to depend linearly on HRP concentration. Methods for the determination of HRP concentration using the chemiluminescence was investigated. HRP could be detected in the range from 100 pmol/L to 100 nmol/L under the optimum condition, H2O2 (10 mmol/L) and MCLA (10 mumol/L) at pH 5.8.

Animals↗

PubMedDate not supplied

Establishment of an alpha-fetoprotein-producing cell line derived from gastric cancer.

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Aged↗

PubMedDate not supplied

Antisense epidermal growth factor receptor delivered by adenoviral vector blocks tumor growth in human gastric cancer.

Epidermal growth factor receptor (EGFR) protein overexpression is commonly found in human gastric cancer, and its gene amplification is known to correlate with poor prognosis in gastric cancer patients. With regard to therapy trials targeting EGFR, it has been reported that stable transfection of EGFR antisense or treatment with antibody against EGFR results in growth suppression of human cancer cells that express high levels of EGFR. We have designed an adenovirus-expressing antisense EGFR and have investigated its effect on the growth of gastric cancer in vitro and in vivo. Following infection with EGFR antisense RNA-expressing adenovirus (Ad-EAS), the cell surface EGFR protein levels of infected cancer cells were markedly reduced, and the in vitro growth of Ad-EAS-infected cells was significantly inhibited relative to control-infected cells in all three gastric cancer cell lines (AGS, KKLS, and MKN28) studied here (P < .0002). In a nude mouse subcutaneous tumor system, in vivo tumor growth of MKN28 was significantly inhibited after Ad-EAS treatment, and inhibition on day 48 was 93% by volume compared with that of untreated controls. These results suggest that an adenoviral vector system targeting the down-regulation of EGFR could be a good candidate for the therapy of gastric cancers that overexpress EGFR.

Adenoviridae↗

PubMedDate not supplied

Subcellular localization of PP5/TFPI-2 in human placenta: a possible role of PP5/TFPI-2 as an anti-coagulant on the surface of syncytiotrophoblasts.

Placental protein 5 (PP5)/tissue factor pathway inhibitor-2 (TFPI-2), a serine proteinase inhibitor, is homologous to tissue factor pathway inhibitor (TFPI) and commonly found in peripheral blood of pregnant woman. Although TFPI is well known to be synthesized primarily in endothelium and to play an important role in regulation of the extrinsic pathway of blood coagulation, the function of PP5/TFPI-2 remains unclear. Our previous report demonstrated that PP5/TFPI-2 expression is ubiquitous and extremely high in growing placental tissue. Using newly generated polyclonal anti-PP5/TFPI-2 antibody, and by immunohistochemistry and immunoelectromicroscopy, we examined precise localization of PP5/TFPI-2 in placenta especially in syncytiotrophoblasts, which had been shown to produce PP5/TFPI-2 mRNA by our previous study using in situ hybridization. Immunoelectromicroscopy revealed PP5/TFPI-2 localizing on the surface of the microvilli and the membrane of endoplasmic reticulum of syncytiotrophoblasts. To confirm the cell surface association of PP5/TFPI-2, placental villi were incubated with heparin and resultant soluble fraction was analysed by Western blotting. Heparin liberating PP5/TFPI-2 from villi suggested that PP5/TFPI-2 might be retained on the microvilli surface through the binding to membrane-anchored proteoglycans such as glypican and/or syndecan family members. We also examined the relationship between the presence of cell layer of syncytiotrophoblasts and the coagulation using clinical specimens, and revealed that the fibrin depositions were specifically observed on the regions lacking syncytiotrophoblasts cell layer in placental villi. Therefore, it is likely that during pregnancy PP5/TFPI-2 may be retained on the surface of placental villi via proteoglycans, and may play an important role to maintain intervillous blood flow and the patency of microvasculature in feto-maternal blood system mediated by the inhibition of serine proteinases involved in the blood coagulation.

Adult↗

PubMedDate not supplied

MR imaging of bilateral adrenal hemorrhage.

Serial magnetic resonance (MR) findings (2 weeks and 7 weeks after onset of symptoms) in a case of bilateral adrenal hemorrhage, complicating severe infection are reported. Spin echo 600/20 images initially showed triangular enlarged adrenal glands with intermediate intensity centers and concentric high intensity outer rings. Spin echo 2,000/60 images showed inhomogeneous intensity. Follow-up MR examination 5 weeks later demonstrated that the hematomas decreased in size, and the signal intensity changes showed resolving hematomas (i.e., increased signal intensity centrally, decreased peripherally).

Adrenal Gland Diseases↗

PubMedDate not supplied

Heterogeneity of immune markers in hemophagocytic lymphohistiocytosis: comparative study of 9 familial and 14 familial inheritance-unproved cases.

PURPOSE: Although immune dysfunction is suspected in patients with hemophagocytic lymphohistiocytosis (HLH), the difference between immune dysfunction in patients with familial erythrophagocytic lymphohistiocytosis (FEL) and familial inheritance-unproved lymphohistiocytosis (FIU) remains unknown. The aim of this study was to determine useful markers to distinguish patients with FEL from those with FIU. PATIENTS AND METHODS: Clinical features and laboratory findings, especially natural killer (NK) cell activity and the relative frequencies of peripheral blood mononuclear cell (PBMC) subsets, and serum levels of interferon-gamma and soluble interleukin-2 receptor were compared in 9 patients with FEL and 14 age-matched patients with FIU. Twenty-seven healthy infants served as controls. The treatment and outcome were also compared for patients with FEL and FIU. RESULTS: Comparison between patients with FEL and FIU revealed significantly lower NK activity in those with FEL (p = 0.03) but failed to show any significant differences in PBMC subsets, except that the percentage of CD3+ T cells was higher in patients with FEL (p = 0.02). CD4- and CD8-dominant phenotypes were characteristic findings in both groups of patients, although increased CD19+ B cells were restricted to patients with FIU. NK activity was deficient (< 5%) in four of the seven patients with FEL tested but in only one of eight patients with FIU. By comparison to values for age-matched controls, the percentages of CD3+, CD3+DR+ and CD45RO+ PBMCs in patients with FEL were significantly high (p < 0.05) and those of CD19+ and CD45RA+ subsets were lower than normal. Among patients with FIU, PBMC subsets included significantly reduced CD3+, CD4+, CD45RA+, and CD4+CD45RA+. In this small series, the outcome of patients with FEL and FIU treated with chemotherapy was not significantly different at the time of evaluation. CONCLUSIONS: These results indicate considerable immune heterogeneity among patients with HLH younger than 2 years. Although NK activity was useful but not diagnostic, determination of PBMC subsets and patterns of cytokine expression was not helpful in distinguishing patients with FEL from those with FIU, suggesting that the immune responses characteristic of these diseases may reflect different triggering factors, including viruses. The impact of this immune heterogeneity on patients' outcome remains to be determined.

Antigens, CD↗

PubMedDate not supplied

Separation of benzodiazepines using cholesterol-modified fused-silica capillaries in capillary electrochromatography.

This paper describes the use of cholesteryl-10-undecenoate as a capillary column modifier for capillary electrochromatography. This bonded phase has shown an entirely different selectivity as compared to octadecyl silica (ODS) phases in reversed-phase liquid chromatography, specifically in the analysis of the benzodiazepines. This difference serves as the basis for the continuing study as well as the present focus on electrochromatography as a relatively new separation technique. To significantly increase the surface area, etching of the inner wall of a 75-micron capillary is performed using ammonium hydrogen difluoride and is then subsequently modified via a silation reaction with triethoxysilane and subjected to hydrosilation with cholesteryl-10-undecenoate to form the bonded phase. The performance of this capillary is compared with unetched cholesterol-modified and bare, fused-silica capillaries. Efficient resolution was noted for the etched capillary showing the effectiveness of the etching process as well as the selective property of the bonded phase.

Benzodiazepines↗

PubMedDate not supplied

Indication of splenectomy for gastric carcinoma involving the proximal part of the stomach.

BACKGROUND/AIMS: The role of splenectomy in the surgical management of gastric carcinoma is controversial and there is no consensus of opinion regarding the therapeutic value of splenectomy. The aim of this study was to search for possible metastasis to lymph nodes in the splenic hilum or along the splenic artery to avoid unnecessary splenectomy and to determine its indication. METHODOLOGY: The clinical records of 204 patients who underwent total gastrectomy combined with splenectomy for gastric carcinomas involving the proximal part of the stomach were analyzed. RESULTS: The incidence of nodal involvement to the splenic hilum and/or along the splenic artery was 49 (24.0%) of 204 gastric carcinomas involving the proximal part of the stomach that underwent combined gastrectomy and splenectomy. The characteristics of gastric carcinoma with metastasis to these nodes included a larger tumor, deeper penetration (T3, 4 tumors), a number of lymph node metastasis, and infiltrative type. In T2 cases, all the tumors with cancerous involvement to these nodes showed intraoperative gross serosal change). When the tumor size was less than 40 mm, nodal metastatic rate to the splenic hilum and/or along the splenic artery was very low. CONCLUSIONS: In conclusion, splenectomy should be conducted in T2 cases with gross serosal change and T3, 4 cases. With regard to tumor size, in the cases with a tumor whose size was less than 40 mm, it is possible to preserve the spleen in most cases. In the near future, splenectomy should be clarified precisely by randomized trials in advanced gastric carcinoma.

Adenocarcinoma, Papillary↗

PubMedDate not supplied

Annexin II overexpression is correlated with poor prognosis in human gastric carcinoma.

Annexins belong to a family of the calcium-dependent phospholipid binding proteins. They are also substrates of receptor tyrosine kinases. Overexpression of Annexin II, which has been reported in various carcinomas, is thought to be associated with cell proliferation, differentiation and cell-cell adhesion in the pathogenesis of carcinoma, but the functions of Annexins have not been fully elucidated. In this study, we investigated the role of Annexin II (p36) and its relationship with c-erbB-2 overexpression in gastric carcinoma. We studied Annexin II expression using Western blot analysis in 8 human gastric carcinoma cell lines and expression of Annexin II and c-erbB-2 using, immunohistochemistry in 153 primary gastric carcinomas. Western blot revealed that Annexin II was expressed in 8 human gastric carcinoma cell lines. It was more strongly expressed in the cell membrane than in the cytoplasm of tumor cells in primary gastric carcinoma tissues. Thirty-three percent of all cases were immunopositive for Annexin II, overexpression of which was more frequent in differentiated type (p = 0.0009), lymph node, metastasis (p = 0.0147) and venous invasion (p = 0.0092). Annexin II and c-erbB-2 overexpression were significantly correlated p = 0.0002) and patients with Annexin II had poorer prognoses (p = 0.0066). Multivariate analysis showed that immunopositivity of both Annexin II and c-erbB-2 was an independent and poor prognostic factor (p = 0.0037). In conclusion, Annexin II was overexpressed in advanced gastric carcinomas and it could contribute to the progression of gastric carcinoma.

Annexin A2↗

PubMedDate not supplied

Detection of malignant thymoma in primary tumor and metastatic lesions using 99mTc-tetrofosmin scintigraphy.

99mTc-tetrofosmin was developed as a myocardial perfusion imaging agent and can also be used to depict tumors. We have experienced five cases of malignant thymoma delineated on 99mTc-tetrofosmin SPECT. In one case significant activity was clearly detected in the primary tumor and metastatic lesions. In quantitative analysis, similar 99mTc-tetrofosmin and 201Tl-chloride uptake ratios were obtained (1.95+/-0.57 versus 2.27+/-0.85, respectively; n.s.). The ability of 99mTc-tetrofosmin to detect malignant thymoma was comparable to that of 201Tl-chloride. Therefore, 99mTc-tetrofosmin might be a useful tracer for the detection of malignant thymoma, although more studies will be required to evaluate its diagnostic accuracy.

Adult↗

PubMedDate not supplied

Studies on metabolism of bromazepam. VI. Reduction of 2-(2-amino-5-bromobenzoyl)pyridine, a metabolite of bromazepam, in the rabbit, rat, and guinea pig.

Three urinary metabolites that were formed by cleavage of the benzodiazepine ring of bromazepam, 2-(2-amino-5-bromobenzoyl)pyridine (ABBP), 2-(2-amino-5-bromo-3-hydroxybenzoyl)pyridine (3-OH-ABBP), and 2-amino-5-bromo-2'-azabenzhydrol (ABAB), were measured in urine of rabbits, rats, and guinea pigs. The major metabolite was 3-OH-ABBP in all animals given bromazepam orally. ABAB was also excreted in major amounts in the guinea pig, but was excreted in minor amounts in the rabbit and rat. Moreover, ABAB was excreted in the urine of all animals given ABBP orally. It may be concluded that ABAB was formed by reduction of the carbonyl group of ABBP. ABBP reduction was catalyzed by NADPH-dependent enzymes occurring in rabbit liver cytoplasm, and rat liver microsomes, and guinea pig liver cytoplasm and microsomes. The reductases were inhibited by sulfhydryl group reagents. The optimum pH of the cytoplasmic enzyme ranged from 7.2 to 7.8, and that of the microsomal enzyme was 6.5. The apparent KM value for the reduction of ABBP by guinea pig liver microsomes was the lowest among all of the liver preparations.

Animals↗

PubMedDate not supplied

[Prospective study of cases of acute viral conjunctivitis in the city of São Paulo. Characterization of enterovirus 70 (acute hemorrhagic conjunctivitis)].

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Adolescent↗

PubMedDate not supplied

Cardiomyopathy and other chronic toxic effects induced in rabbits by doxorubicin and possible prevention by coenzyme Q10.

Cumulative dose-dependent toxic effects, particularly cardiomyopathy, induced by doxorubicin and the possible prevention by coenzyme Q10 (CoQ10) were studied in rabbits. In rabbits given doxorubicin alone, there was considerable body weight loss, alopecia, pancytopenia, significant increase in serum creatine phosphokinase and LDH, and ECG changes characterized by tachycardia, flat and inverted T wave, the premature ventricular contractions. In rabbits given doxorubicin plus CoQ10, the only change was pancytopenia. In rabbits treated with doxorubicin alone, the most prominent histologic changes were observed in mitochondria of myocytes, and these changes were characterized by loss of outer membrane, disarrangement of cristae, and formation of numerous concentric lamellae. In addition to mitochondrial changes, there were numerous vacuolizations and extensive depositions of both electron-dense and membranous laminated bodies in the sarcoplasm and disarrangement of Z-band and filamentous changes of myofibrils. Numerous vacuolizations in the capillary endothelial cytoplasm in the myocardium were also conspicuous. On the other hand, few significant morphologic changes were seen in the nuclei of myocytes. There were few ECG and histopathologic changes in rabbits treated with both doxorubicin and CoQ10. These findings suggest that the cardiomyopathy of doxorubicin may be prevented or is at least inhibited by CoQ10. The mechanisms of both doxorubicin-induced cardiomyopathy and its prevention by CoQ10 are discussed.

Animals↗