Expression of survivin in esophageal cancer: correlation with the prognosis and response to chemotherapy.

Survivin, a new member of the inhibitor-of-apoptosis (IAP) family, has been reported to be expressed in many cancers but not in differentiated normal tissue. Its expression in esophageal cancer, however, has not been reported. We investigated 51 esophageal cancers and their adjacent normal epithelial tissues for mRNA expression of survivin by RT-PCR. The survivin expression in esophageal cancer tissue was significantly higher than that in normal esophageal tissue (0.211 +/- 0.226 vs. 0.057 +/- 0.135, p < 0.0001). pN4 tumors had significantly higher survivin expression than the pN0-3 tumors (p = 0.0093). Fourteen patients with advanced esophageal cancer had received chemotherapy prior to surgery. The survivin expression in the cancer tissue in patients who achieved a partial response (PR) was significantly lower than that in patients with no change (NC) and in patients with progressive disease (PD; 0.099 +/- 0.134 vs. 0.320 +/- 0.222, p = 0.0434). The median survival for patients with high survivin expression (9.0 months) was less than that for patients with low survivin group expression (30.0 months, p = 0.0023). Survivin expression was one of the significant predictors of survival on univariate analysis (hazard ratio 2.471; 95% confidence interval 1.104-5.533). The results suggest that survivin expression may provide prognostic information in patients with esophageal cancer.

Effectiveness of the Kampo kami-shoyo-san (TJ-24) for tremor of antipsychotic-induced parkinsonism.

The aim of the present study was to evaluate the effectiveness of the Kampo medication kamishoyo-san against tremor due to antipsychotic-induced parkinsonism. Kami-shoyo-san consists of several medicinal herbs that are known in traditional Chinese medicine to be effective in treating Parkinson's disease and convulsions. We gave kami-shoyo-san to eight patients at Higashiowari National Hospital who were exhibiting tremor, a symptom of antipsychotic-induced parkinsonism. The tremor was evaluated on a five-point scale before and after the administration of kamishoyo-san, and the findings were compared statistically. The results showed a stastistically significant reduction in the tremor after the administration of kami-shoyo-san (P<0.01), with 62.5% of patients showing an improvement of one point or more. The tremor did not worsen in any of the patients, and none complained of side-effects. Thus, kami-shoyo-san appears to be effective against the tremor from parkinsonism. Kami-shoyo-san consists of 10 medicinal herbs, including Radix Bulpleuri, Radix Paeoniae, Radix Angelicae Sinensis, and Radix Glycyrrhizae, which are effective in treating disturbances in muscular movement according to TCM theory. Of the 10 herbs contained in kami-shoyo-san, we believe the abovementioned ones are particularly effective in helping to reduce the tremor associated with parkinsonism.

Crystal transformation from anhydrous alpha-maltose to hydrous beta-maltose and from anhydrous trehalose to hydrous trehalose.

Anhydrous sugars such as maltose and trehalose are useful for making dry powder of foods and liquids. The crystal-transformation rate of maltose and trehalose were investigated under humid conditions and by kneading. The enthalpy for solubilization was 7.0 kJ/mol for the anhydrous maltose. The crystal-transformation rate of anhydrous alpha-maltose to hydrous beta-maltose depended on the temperature at 75% humidity. However, that of anhydrous trehalose did not depend on the temperature, and transformation was very rapid. An anomeric change to maltose and no such change to trehalose might have caused this. The activation energy of crystal transformation was 79 kJ/mol for maltose and zero for trehalose. The rate of crystal transformation of anhydrous maltose while kneading depended on the purity of the anhydrous alpha-maltose and the amount of water present. This crystal transformation rate fitted the Avrami equation.

High frequency of the MAGE-1 gene expression in hepatocellular carcinoma.

The MAGE-1 gene, originally isolated from a human melanoma cell line, directs the expression of a potential tumor-rejection antigen, MZ2-E. This antigen is recognized by autologous cytotoxic T lymphocytes in association with a major histocompatibility complex class I molecule (HLA-A1), and has provided a basis for specific immunotherapy for melanoma patients. Here we show a high frequency of expression of the MAGE-1 gene in hepatocellular carcinoma (HCC). We examined the expression of the MAGE-1 gene in cell lines originated from hepatoma cells and in tumor and nontumor tissues of livers with HCC by reverse-transcription polymerase chain reaction (RT-PCR) and subsequent Southern blotting. MAGE-1 messenger RNA (mRNA) was expressed in three of four HCC cell lines (75%) and in 16 of 20 (80%) resected HCCs though none was detected in nontumor tissues. The high frequency expression of MAGE-1 gene in HCCs suggests the possibility as a target for tumor-specific immunotherapy for HCC patients.

Rotenone, a mitochondrial NADH dehydrogenase inhibitor, induces cell surface expression of CD13 and CD38 and apoptosis in HL-60 cells.

We previously demonstrated that the mitochondrial NADH dehydrogenase subunit 2 (ND2) gene was overexpressed in human acute myelogenous leukemia (AML) cells. Since this finding suggested that ND2 gene expression was related to myeloid differentiation, we here investigated the effects of rotenone, a specific NADH dehydrogenase inhibitor, on HL-60 cell growth, differentiation and death. Fifty nM rotenone inhibited the growth of HL-60 cells and caused an increase in the cell population in the G(2) +M phase. In the quantitative comparison of myeloid antigen, the expression of CD13 and CD38 were relatively increased in the rotenone-treated cells. These findings suggest that the inhibition of NADH dehydrogenase changes the cell cycle and induces some specific surface antigens of HL-60 cells. On the other hand, the expression of ND2 gene remained unchanged after the rotenone treatment, suggesting the rotenone-mediated mitochondrial inhibition did not affect the mitochondrial gene expression. Five mu M rotenone strongly inhibited the cellular proliferation. Electron microscopy and an electrophoretic analysis of DNA showed that the majority of the HL-60 cells were induced into typical apoptosis within 24-48 hours. On the basis of this and other studies, we believe that mitochondrial function is directly involved in both cellular differentiation and apoptotic cell death.

Serial chromosomal change during the progression of hepatocellular carcinoma.

We investigated the chromosomal loss pattern of a primary large nodule and one of the disseminated intrahepatic small nodules of hepatocellular carcinoma in one patient using RFLP analysis. Using D17S30 probe mapping to 17p, loss of heterozygosity was observed both the main nodule and the disseminated nodule, however, using IGLV probe mapping to 22q, loss of heterozygosity was observed only in the disseminated nodule. This is the case in which serial loss of chromosome in the progression on hepatocellular carcinoma was highly suggested.

Tumor necrosis factor production by rat Kupffer cells-regulation by lipopolysaccharide, macrophage activating factor and prostaglandin E2.

Tumor necrosis factor (TNF) is considered to be deeply involved in the hepatocyte damages in severe hepatitis. To delineate which mediators are involved in the production of TNF in vivo, we examined regulatory mechanisms of the production of TNF by rat Kupffer cells using a variety of mediators. Lipopolysaccharide (LPS) markedly induced TNF production by Kupffer cells. Kinetic studies revealed a rapid release of TNF within 3-4 hrs after the addition of LPS to the culture medium. Spleen cell derived-macrophage activating factor prepared from rat spleen cells did not by itself induce the production of TNF. However, the presence of a small amount of the factor during or before exposure to LPS induced higher levels of TNF, suggesting that macrophage activating factor had a priming effect. Recombinant human interferon-gamma and recombinant human granulocyte-macrophage colony stimulating factor, the natural types of which are components of the macrophage activating factor, displayed similar effects. Prostaglandin E2 (PGE2) and dexamethasone both inhibited LPS-induced TNF production in a dose dependent manner. Indomethacin, a cyclooxygenase inhibitor, increased LPS-induced TNF production. Interestingly, a combination of PGE2 and indomethacin inhibited TNF production more strongly than PGE2 alone, suggesting that the simultaneous treatment with PGE2 and indomethacin decreases liver damage in severe hepatitis rather than PGE2 alone. In addition, PGE2 pretreatment reduced the response to the newly added PGE2, suggesting the presence of a desensitization mechanism in the PGE2 receptor system. These findings suggest that spleen cell-derived macrophage activating factor and bowel-derived LPS take important parts in TNF production through the portal blood in the liver in vivo.

[Bladder cancer in spinal cord injury patient. A case report].

A case of bladder cancer following spinal cord injury is reported. A 57-year-old male with L1 incomplete paraplegia was referred to our hospital under the diagnosis of bladder tumor with gross hematuria in November, 1994. Radical cystectomy, rt nephroureterectomy (nonfunctioning kidney) and lt ureterocutaneostomy were carried. Histopathological study revealed squamous cell carcinoma (well differentiation) and transitional cell carcinoma (G3), stage pT3b. He has been doing well without recurrence for 4 months after operation.

Presence of C5a-potentiating activity in the plasma of a patient with Kimura's disease.

To determine whether the response to the active complement fragment C5a in polymorphonuclear leukocytes (PMN) obtained from a patient with Kimura's disease is similar to that in normal subjects, we evaluated superoxide anion (O2-) generation after stimulation with C5a. The patient's PMN produced more O2- than did those from healthy controls after stimulation with human C5a in vitro. The response returned to normal with the improvement in clinical indicators after initiation of steroid administration. We conducted the following experiments with the plasma of this patient during the active disease stage to establish the presence of a humoral factor that potentiated the C5a-response of PMN. Incubation of PMN from normal controls having the same blood type as the patient at 37 degrees C for 2 h with the patient's plasma at the active disease stage revealed that the cells generated abundant O2- after stimulation with C5a. Incubation of normal PMN with the patient's plasma during the inactive disease stage showed no potentiated response to C5a. This activity was lost after incubation at 56 degrees C for 30 min. Coincubation of PMN with methylprednisolone up to 100 micrograms/ml did not suppress this activity. Although the plasma concentration of C5a at the active stage was mildly elevated (13 ng/ml), it was below the limit of detection (< 10 ng/ml) at the inactive stage. These results suggest the presence of a heat-labile, humoral factor in the patient's plasma that upregulated the response of PMN to C5a and that was not suppressed by in vitro treatment with a steroid. This factor may influence the acute inflammatory reaction in this disorder.

Different subsets of CNS neurons express different glycosaminoglycan epitopes on large perineuronal proteoglycans.

Proteoglycans are known to occur in the central nervous tissue, but their function is not well understood. We made a biochemical study of four perineuronal antigens on different subsets of rat brain neurons and found that three antigens recognized by antibodies against glycosaminoglycan epitopes were large proteoglycans. Molecular masses estimated by immunoblotting were 700 kDa for 473, 376 and 1B5 antigens and 600 kDa for the 374 antigen. Reactivities of the antigens on immunoblots to monoclonal antibodies (MAbs) 473 and 376 were lost, and that to MAb 1B5 uncovered, after chondroitinase ABC treatment as in the brain sections. The elution profiles from ion-exchange and gel-filtration column chromatography of 473, 376 and 1B5 antigens were quite similar, but those of 374 antigen were slightly different. The immunoaffinity-purified 473 antigen migrated at 700 kDa on sodium dodecylsulfate gel electrophoresis, and chondroitinase ABC treatment decreased the molecular mass to 600 kDa. The 473 antigen was recognized by MAbs 376 and 1B5 although these MAbs also reacted with 473-negative 700 kDa molecules. These results suggest that neuronal subset-specific glycosylation may occur on the 700 kDa proteoglycans, and that the glycosaminoglycan structures may be involved in the pericellular diversity of CNS neurons.

Human monoclonal antibodies from a patient with primary biliary cirrhosis that recognize two distinct autoepitopes in the E2 component of the pyruvate dehydrogenase complex.

Peripheral B lymphocytes from a patient with primary biliary cirrhosis were infected with Epstein-Barr virus, and Epstein-Barr virus-transformed B lymphocytes producing large amounts of IgG antibodies to pyruvate dehydrogenase complex were selected, expanded and fused with the human-mouse heteromyeloma cell line F3B6. The resulting Epstein-Barr virus-transformed B-cell hybrids were repeatedly cloned by limiting dilution, and three stable hybridoma clones producing human monoclonal antibodies to pyruvate dehydrogenase complex were generated. These monoclonal antibodies, designated M18GP8, M37GP11 and M82GP8, specifically bound to pyruvate dehydrogenase complex, and their dissociation constant with pyruvate dehydrogenase complex was calculated to be 2.4 x 10(-11), 2.3 x 10(-10) and 2.6 x 10(-11) mol/L, respectively. These three monoclonal antibodies stained the mouse stomach/kidney cryostat sections in a typical immunofluorescence pattern of antimitochondrial antibody. Furthermore, the enzymatic activity of pyruvate dehydrogenase complex was almost completely inhibited by the three monoclonal antibodies. Western blotting analysis revealed that M18GP8 and M82GP8 reacted with only pyruvate dehydrogenase complex-E2 in contrast to M37GP11, which reacted with both pyruvate dehydrogenase complex-E2 and protein X. The binding of monoclonal antibody M37GP11 to solid-phase pyruvate dehydrogenase complex was partially inhibited by two different synthetic peptides corresponding to both the inner and outer lipoyl-binding domains of pyruvate dehydrogenase complex-E2. These monoclonal antibodies, which are the first human monoclonal antibodies to pyruvate dehydrogenase complex generated from a patient with primary biliary cirrhosis, will be a valuable tool for studying the B-cell autoepitopes in PDC and the mechanism of autoantibody production in primary biliary cirrhosis.

[Interferon therapy after ablation of Kent bundle for a patient with chronic hepatitis type B complicated with WPW syndrome].

Cardiotoxicity of interferon-alpha or gamma, such as fatal arrhythmia and myocardial infarction, has been reported. Therefore cardiotoxicity of interferon should be seriously considered before administration for patients with a pre-existing heart disease. We treated a patient with chronic active hepatitis type B, coexisted with Wolff-Parkinson-White syndrome, who has had frequent attacks of paroxysmal atrial fibrillation. To prevent the occurrence of fatal arrhythmia with an interferon therapy in this patient, we performed radiofrequency catheter ablation of the Kent bundle. After the successful ablation, we could safely administered recombinant interferon alpha-2b for chronic hepatitis type B.

Frequent loss of heterozygosity on chromosome 22 in hepatocellular carcinoma.

We investigated 24 hepatocellular carcinomas in Japan to find loss of heterozygosity with 15 polymorphic DNA markers that detect allelic losses at specific chromosome loci. Loss of heterozygosity on chromosomes 10q, 17p and 22q was detected in 3 of 12 (25%), 9 of 21 (43%) and 5 of 15 (33%) informative cases of hepatocellular carcinoma, respectively. This is the first report of loss of heterozygosity on chromosome 22q in hepatocellular carcinoma; the newly recognized common chromosome loss was considered to exist between D22S9 and D22S10 on 22q11. On the basis of this and other studies, we believe it is likely that such a chromosome loss in hepatocellular carcinoma is a signal for malignant transformation and that loss of unknown genes on chromosomes 10q, 17p and 22q may contribute to tumor progression in hepatocellular carcinoma.

Electron microscopic localization and expression on cultured cells of a novel surface-associated molecule in the rat central nervous system.

Subcellular localization and in vitro expression of a newly raised monoclonal antibody 374 antigen, which outlines a subset of neurons in various regions of the rat CNS, were studied. Electron microscopy revealed that the immunoreactivity is associated with the outer surfaces of neuronal subsets, the surfaces of glial cell processes facing such neurons and less frequently, with the Golgi apparatus and inner cell surfaces. The monoclonal antibody also stained part of the cytoplasm of a population of dendrites. In order to test whether the antigen originates from neurons, we examined the expression of the antigen in cultured cortical cells. Double immunofluorescence staining with neuron-specific enolase and glial fibrillary acidic protein revealed that the antigen is expressed on a subset of neuronal cells and to a lesser extent on glial cells. The antigen is considered to be expressed on the outer surface because the cells were stained by the monoclonal antibody irrespective of whether they were alive or fixed. The monoclonal antibody 374 antigen may be a novel neuronal surface molecule which plays a role in neuron-neuron or glial-neuron interactions in the CNS.

[Loss of heterozygosity in hepatocellular carcinoma].

Several genetic events are necessary for a cell to become malignant. Some of these events are activations of protooncogenes and loss of normal functions of tumor suppressor genes. Recently loss of heterozygosity at specific loci on some chromosomes have been observed in several tumors. Loss of heterozygosity in the tumor suggests that a certain tumor suppressor gene may reside near the locus of the probe by which the loss of heterozygosity was demonstrated. In hepatocellular carcinoma, loss of heterozygosity on chromosome 1p, 4q, 5q, 10q, 11p, 13q, 16q, 17p and 22q are observed. Inactivation of tumor suppressor genes on these chromosomes may play important roles in tumorigenesis or progression of hepatocellular carcinoma.

[5-fluorouracil + low-dose leucovorin and cisplatin sequential chemotherapy with dipyridamole for advanced nonresectable squamous cell carcinoma of the lung: a case report].

A 62-year-old man diagnosed as Stage IIIB advanced non-resectable squamous cell carcinoma of the lung was treated with a sequential combination of 5-fluorouracil (5-FU) and cisplatin (CDDP), with concurrent administration of leucovorin and dipyridamole as a biochemical modulator for 5-FU. After 3 cycles, the mass reduced in size more than 70% in CT scan and the patient underwent a thoracotomy. Histologically, the primary lesion was completely necrotized and of the 10 metastatic regional lymph nodes, only one lymph node contained a small amount of viable cells and 3 additional cycles were conducted. The patient is still alive 30 months after initial chemotherapy. This regimen appears to be potentially useful for non-small-cell lung cancer and warrants further clinical study.

[Effective continuous hemofiltration and plasma exchange for the treatment of subacute type fulminant hepatic failure].

By combination of continuous hemofiltration (CHF) with plasma exchange therapy we successfully treated a patient with subacute type fulminant hepatitis to keep her consciousness alert. The patient was a 55-year-old woman who admitted because of severe jaundice. On the 51st day after the onset she had consciousness disturbance and was transferred to our hospital. We started the therapy of CHF and plasma exchange at the patient's hepatic coma grade 4. On the 5th day her consciousness level recovered to grade 2 and we could keep the level for almost 2 weeks. This combination therapy seemed good not only for the improvement of consciousness of patients in hepatic coma but also to support the hepatic function of almost ahepatic patients.

[A patient with liver cirrhosis manifesting various symptoms including cerebellar ataxia due to germanium intoxication].

A patient with hepatitis B virus-associated cirrhosis manifested various symptoms such as anemia, renal damage and neurological signs including cerebellar ataxia due to long-term administration of germanium-containing food. The patient was a 40-year-old male who had taken germanium containing mineral cheese for 26 months after he was diagnosed as having cirrhosis. Twenty four months after beginning to take the mineral cheese, he began manifesting paresthesia of the extremities, dysarthria and gait ataxia. Laboratory findings revealed anemia and renal damage. Biopsy of the peripheral nerve revealed loss of the large sheathed nerve, a characteristic feature of germanium intoxication. A high concentration of germanium (GeO2) was detected in patient's hair and urine. Cerebellar ataxia was characteristic in this patient, which was not reported in the previous papers.