Interaction of bloom-syndrome cellular cancer antigens with sera of malignant-lymphoma patients - an immunological and cytogenetical study

We have analyzed chromosome abnormalities in 32 B-cell lymphoma (B-ML), 29 T-cell lymphoma (T-ML) and 29 adult T-cell leukemia/lymphoma (ATL) patients and tested the reactivity of Bloom syndrome (BS) derived malignant lymphoma (ML) antigen with the sera in these patients with immunofluorescence (IF) and Western blotting (WB) protocols. Among the cases analyzed, 14q32 abnormalities were observed in 11 (B-ML), 8 (T-ML) and 10 (ATL) cases; these changes were accompanied by complex numerical and structural abnormalities. In the IF and WB analyses, the reactivity of BS ML antigen and ML sera showed high positivity in over 85% of ML cases and appeared to clearly raise the number of successful diagnoses as compared to the chromosome finding, giving 50-60% success in identifying ML. WB analyses demonstrated common ML antigen band at 97 KD (B-ML, T-ML and ATL). DNA analysis from BS ML antigen cells (BS-SHI-4M ML) showed rearranged fragments in both T-cell receptor (TCR alpha3 and immunoglobulin joining region (I(g)J(H) genes. The present findings of rearrangements of TCR alpha3 and I(g)J(H) in BS-SHI-4M ML antigen cells possibly support the common reactivity of BS ML antigen to B- and T-ML and ATL sera. The possible role of immunoglobulin heavy chain variable region (I(g)V(H) gene mutation and ML antigen expression is discussed.     

Intracellular activation and cytotoxic action of RS-1541 against cultured human tumor cells

RS-1541, an acyl-derivative of rhizoxin (Fig. 1), is a potent antitumor compound. This agent showed cytotoxicity in vitro on some cultured human tumor cells, although it was less potent than rhizoxin. Rhizoxin exhibited antitumor effects by inhibiting the polymerization of tubulin, whereas RS-1541 did not inhibit tubulin polymerization in vitro. However, cell cycle analysis in vivo showed that the two agents had the same mode of action. The cytotoxicity of RS-1541 was enhanced when the initial cell density of the cells was increased. The cytotoxicity was also enhanced when the membrane fraction of St-4 cells, which were the most sensitive to RS-1541 among the cell lines tested, was added to the target cells. When St-4 cells were incubated with [¹⁴C]-RS-1541, significant amounts of [¹⁴C]-rhizoxin were produced within the cells. Further fractionation of the crude membrane showed that the activity that enhanced the cytotoxicity of RS-1541 (RS-1541-enhancing activity) belonged to the mitochondrial-lysosomal fraction, not to the microsomal fraction. Both the enhancing activity and the activity that converting [¹⁴C]-RS-1541 to [¹⁴C]-rhizoxin (RS-1541-converting activity) were inhibited by treatment with chloroquine, an inhibitor of lysosomal function. Cholesterol esterase derived fromCandida cylindracea had RS-1541-enhancingand-converting activities. These data suggest that RS-1541 exerts its cytotoxic action after being converted to rhizoxin within the cells by a lysosomal enzyme such as cholesterol esterase.Abbreviations DMSO Dimethylsulfoxide - PBS(-) Ca²⁺ Mg²⁺-free phosphate-buffered saline - HCO60 hydrogenated castor oil polyethylene glycor ether - DMA dimethylacetamide - RSB reticulocyte standard buffer, consisting of 10mM NaCl, 1.5 mM MgCl₂, and 10 mM TRIS-HCl, (pH 7.4) - TLC thin-layer chromatography - ara-C 1--D-arabinofuranosylcytosine - LDL low-density lipoprotein     

Anterior Transhepatic Approach for Isolated Resection of the Caudate Lobe of the Liver

RID=" ID=" <E5>Correspondence to:</E5> J. Yamamoto, M.D.     

Leptin effects on feeding-related hypothalamic and peripheral neuronal activities in normal and obese rats.

We investigated the effects of leptin on central and/or peripheral feeding-related neuronal networks in Wistar male rats either normal (350-450 g) or Zucker obese (500-800 g). Low doses (1-10 pg) of leptin inhibited glucose-sensitive vagal hepatic afferent discharges and facilitated sympathetic efferent discharges to brown and white adipose tissue. Most (40-75%) neurons in the arcuate nucleus were significantly inhibited by superperfusion with leptin (0.1 nM-10 pM) under in vitro conditions. In anesthetized animals, leptin was applied electrophoretically to single hypothalamic neurons. Both glucose-sensitive neurons (GSNs) and non-GSNs in the feeding center (LHA) were significantly inhibited. Most glucoreceptor neurons in the satiety center (VMH) were significantly excited. Their depolarization was confirmed by activation of Na+ and K+ channels by 10(-11) M leptin using the perforate blind patch-clamp method. Although leptin excited GSNs in the parvocellular part of the paraventricular nucleus, the effects of leptin on such neuronal activity were slight or absent in Zucker obese rats. These results suggest that the feeding-suppression effects of leptin are mediated by its effects on signal transduction through both the central and the peripheral nervous systems.     

Diurnal variation in neutrophil function

Neutrophil functions, including chemotaxis, reactive oxygen species (ROS)-producing capacity of neutrophils, and serum opsonic activity were investigated in 9 young healthy male volunteers. Venous blood of these volunteers was obtained under standardized conditions at 4-h intervals over a 24-h span. Neutrophil chemotaxis was evaluated by a modified Boyden technique, ROS-producing capacity of neutrophils and serum opsonic activity were measured by a simultaneous multiple measurement system based on luminol-dependent chemiluminescence and indicated by peak height and peak time. ROS-producing capacity of neutrophils and serum opsonic activity were activated in the daytime, and decreased from night to morning. There were negative correlations between the peak time of the luminol-dependent chemiluminescent response, neutrophil number (p<0.01) and segmented neutrophil number (p>0.01). On the other hand, no significant correlations were noted between serum opsonic activity and IgG, IgA, IgM, C3 or C4. In contrast, the peaks of neutrophil chemotaxis were at the wake-up time (6:00a.m.) and in the evening (6:00p.m.). This study indicates that diurnal variation of neutrophil function exists.     

Delayed image of iodine-123 iomazenil as a relative map of benzodiazepine receptor binding: The optimal scan time

Delayed single-photon emission tomograpic (SPET) images after an intravenous bolus injection of iodine-123 iomazenil have been used as a relative map of benzodiazepine receptor binding. We determined the optimal scan time for obtaining such a map and assessed the errors of the map. SPET and blood data from six healthy volunteers and five patients were used. A three-compartment kinetic model was employed in simulation studies and analyses of actual data. The simulation studies suggested that, in the normal brain, the scan time at which a single SPET image best represented the relative receptor binding was 3.0–3.5 h post-injection. This finding was supported by actual data from the volunteers. The simulation studies also suggested that the optimal scan time was not greatly changed by the variability of the input functions, and that the error in the SPET image contrast in the vicinity of the optimal scan time was not increased by changes in the tracer kinetics in the entire brain. The SPET image contrast in the patients at 3.0 h post-injection agreed well with the reference receptor binding estimated by kinetic analysis, with a mean error of 3.6%. These findings support the use of a single SPET image after bolus injection of [¹²³I]iomazenil as a relative map of benzodiazepine receptor binding. For this purpose, a SPET scan time of 3.0-3.5 h post-injection is recommended.     

Oxygen uptake and carbon dioxide elimination during epinephrine-induced arrhythmias in humans

Key words  cardiac arrhythmias - oxygen uptake - carbon dioxide elimination     

Curative Resection of Microgastrinomas Based on the Intraoperative Secretin Test

The intravenous secretin injection test (secretin test) has been used for the differential diagnosis of gastrinoma. In this study we report that the intraoperative secretin test (IOS test) is also useful for determining the extent of curability in patients with Zollinger-Ellison syndrome (ZES). Twelve patients with ZES underwent surgical exploration and the IOS test. The results of the IOS test were obtained by rapid radioimmunoassay of the serum gastrin level (IRG) within 60 minutes. The test was diagnosed as negative when the maximum increase of serum IRG was less than 80 pg/ml and also less than 20% of the basal serum IRG level. Three of the twelve patients underwent pancreatoduodenectomy (PD), and two patients underwent distal pancreatectomy. Extirpation of duodenal tumors with dissection of regional lymph nodes was performed in seven patients. In two of the seven patients the IOS test remained positive after extirpation of the duodenal tumors and the dissection of regional lymph nodes. In one patient PD was performed on the basis of the positive results, and the IOS test became negative after PD. In the other patient, two tiny metastatic liver tumors were identified and were resected, but the IOS test did not become negative. We closed the abdomen in 11 patients when we obtained negative results from the IOS test. The results of the IOS test were almost identical to the data obtained by the standard assay postoperatively. The serum IRG levels of all but one patient fell to the normal level, and the secretin test became negative postoperatively. The IOS test is thus useful and indispensable for curative resection of microgastrinomas in patients with ZES.     

Increased expression of angiogenin in hepatocellular carcinoma in correlation with tumor vascularity.

PURPOSE: Neovascularization is known to be one of the major characteristics of human hepatocellular carcinoma (HCC). Angiogenin (ANG), originally discovered in a human colon cancer cell line, is a liver-derived polypeptide that shows strong angiogenic activity in vivo. However, the role of ANG on the development of HCC remains unknown. The present study was designed to examine the implication of ANG in the neovascularization of human HCC. EXPERIMENTAL DESIGN: Forty-one HCC patients who had undergone conventional celiac angiography were used in this study. ANG protein expression and microvessel density (MVD) in HCC specimens obtained by liver biopsy or surgical resection were examined by immunohistochemistry, and the levels were quantified by the KS-400 image analyzing system. ANG mRNA expression in liver tissues was evaluated by in situ hybridization. Serum ANG concentrations were measured by an ELISA. Survival rates were calculated using the Kaplan-Meier method. RESULTS: Immunohistochemistry and in situ hybridization showed greater increments of ANG protein expression and mRNA expression, respectively, in HCC tissues than in the surrounding nontumorous tissues. MVD within tumorous tissues increased according to dedifferentiation of the histological grade of HCC, showing a significant correlation (r = 0.877, P = 0.0009) with ANG expression levels. Mean +/- SD serum ANG levels of healthy subjects and chronic hepatitis (CH) patients were 362.3 +/- 84.1 ng/ml and 331.9 +/- 133.8 ng/ml, respectively, with no significant difference. Serum ANG levels of liver cirrhosis patients (242.4 +/- 126.9 ng/ml) were lower than those of healthy subjects or CH patients and decreased as the fibrosis grade advanced. In HCC patients, despite the cirrhotic background, serum ANG levels increased as the tumor vascularity increased (197.8 +/- 64.9 ng/ml for hypovascular, 326.7 +/- 148.6 ng/ml for hypervascular, and 405.0 +/- 121.3 ng/ml for very hypervascular), in good accordance with histological grading, and significantly decreased (P = 0.015) after successful treatment with transcatheter arterial embolization or percutaneous ethanol injection. HCC patients were conventionally divided into two groups according to the serum level of ANG, those with values higher than the mean level (332.9 +/- 143.8 ng/ml) and those with values lower than the mean,; the 5-year survival rate of the latter group was determined to be significantly higher than that in the former group. CONCLUSIONS: These results suggest that ANG is one of the neovascularization defining factors of HCC. Thus, measuring serum ANG may assist in monitoring the disease, and targeting ANG may provide a new strategy for treating advanced HCC.