Subcutaneous seeding of hepatocellular carcinoma following fine-needle aspiration biopsy.
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Biomedical subjects
Publications and source records attributed to H Isobe.
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Expression vectors based on DNA or plus-stranded RNA viruses are being developed as vaccine carriers directed against various pathogens. Less is known about the use of negative-stranded RNA viruses, whose genomes have been refractory to direct genetic manipulation. Using a recently described reverse genetics method, we investigated whether influenza virus is able to present antigenic structures from other infectious agents. We engineered a chimeric influenza virus which expresses a 12-amino-acid peptide derived from the V3 loop of gp120 of human immunodeficiency virus type 1 (HIV-1) MN. This peptide was inserted into the loop of antigenic site B of the influenza A/WSN/33 virus hemagglutinin (HA). The resulting chimeric virus was recognized by specific anti-V3 peptide antibodies and a human anti-gp120 monoclonal antibody in both hemagglutination inhibition and neutralization assays. Mice immunized with the chimeric influenza virus produced anti-HIV antibodies which were able to bind to synthetic V3 peptide, to precipitate gp120, and to neutralize MN virus in human T-cell culture system. In addition, the chimeric virus was also capable of inducing cytotoxic T cells which specifically recognize the HIV sequence. These results suggest that influenza virus can be used as an expression vector for inducing both B- and T-cell-mediated immunity against other infectious agents.
In recent years, it has become clear that various extracellular substrates play an important role in the process of invasion and metastasis of malignant tumors. In a previous study, we reported that the serum concentration of laminin in renal cell carcinoma and prostate cancer is elevated, and furthermore, is remarkably high in patients with metastasis. However, in bladder cancer, this level is elevated only in those with metastasis, indicating that the serum concentration of laminin is useful as a marker for metastasis or clinical progression of the malignant tumor. In the present study, we measured the serum concentration of type IV collagen 7S-domain, which, as laminin constructs the basement membrane, and compared the result with the serum concentration of laminin, to evaluate the clinical significance of the findings. The serum concentration of type IV collagen 7S-domain was not elevated significantly in any tumor type, although the serum concentration of laminin was elevated in patients with renal cell carcinoma, prostate cancer and bladder cancer with metastasis. There were no significant differences in the serum concentration of type IV collagen 7S-domain in any group with metastasis.
A 58 year old man with arthralgia had been treated with antiinflammatory drugs since November, 1987. On March 1988, he was admitted to a local hospital because of jaundice. On April 18, exploratory laparotomy was done to look for the cause of continued jaundice. Macro-pathological and micropathological findings of the liver were compatible with those of intrahepatic cholestasis. On April 29, he was transferred to Kyushu University Hospital because of persistent jaundice. Physical examination showed jaundice, tachypnea and low grade fever. Laboratory examination showed elevated level of serum bilirubin, marked hypoxia and endotoxemia. Plain chest radiogram revealed ground-glass appearance in bilateral lung fields. He was diagnosed to be complicated with adult respiratory distress syndrome (ARDS). Glucocorticoid therapy and oxygen administration were started immediately. But pulmonary fibrosis developed and he died of respiratory failure. Histological findings of lung necropsy specimen were compatible with those of ARDS. These data strongly suggested that endotoxemia and reticuloendothelial dysfunction due to intrahepatic cholestasis had played important roles to develop ARDS.
To assess the accuracy of the bronchoscopic DNA content analysis, samples of non-small-cell lung carcinomas (NSCLC) were investigated by means of flow cytometry. Samples were dissociated using the detergent Triton X-100. In 58 NSCLC cases, 39 (67%) had DNA aneuploid tumors. We compared the DNA indices of bronchoscopic brushing samples with 21 corresponding surgical samples. In 16 (76%) cases, DNA ploidy of both bronchoscopic and surgical samples were in concordance. In 3 (14%) cases, both bronchoscopic and surgical sample showed DNA aneuploidy, but the number of the DNA aneuploid stem cell lines was different. The cause of these differences was ascribed to the intratumor DNA heterogeneity. In 2 (10%) cases, the bronchoscopic sample showed DNA diploidy, but the surgical sample showed DNA aneuploidy. In these cases, tumor cells obtained by bronchoscopic brushing were so few that the small DNA aneuploid peak was undetectable in the DNA histogram. But the tumor DNA ploidy was evaluated correctly in 90% of 21 cases using bronchoscopic samples. Consequently, despite some drawbacks, the DNA ploidy diagnosis using bronchoscopic samples in this relatively small study, was almost as reliable as surgical samples.
To evaluate the usefulness of contrast material-enhanced ultrasound (US) in detection and treatment of hepatocellular carcinoma (HCC), carbon dioxide was injected as a contrast agent into the hepatic artery in 22 patients with HCC. Plain US had enabled detection of 24 HCC nodules in these patients. Contrast material-enhanced US enabled detection of seven additional nodules, which were confirmed as HCC by means of fine-needle aspiration biopsy performed under guidance with contrast-enhanced US. Six of these seven nodules were detected incidentally during examination of other suspected HCC nodules. Five of the seven nodules were treated with percutaneous ethanol injection (PEI) performed under guidance with contrast-enhanced US; the two other nodules were resected. Contrast-enhanced US made the HCC lesions visible for 15-60 minutes, sufficient time to mark the nodule with an iodized oil-ethanol solution for PEI. Because contrast-enhanced US enabled detection of additional nodules and performance of PEI in lesions not detected with plain US, it may help improve the treatment of HCC.
A total of 92 patients with small-cell lung cancer (SCLC) were randomized to receive cisplatin (80 mg/m2, day 1)/etoposide (100 mg/m2, days 1, 3, 5) (PE) or cisplatin (80 mg/m2, day 1)/etoposide (100 mg/m2, days 1, 3, 5)/ifosfamide (2 g/m2, days 1, 2, 3) (PEI) combination chemotherapy. After 2 courses of chemotherapy, patients with limited disease (LD) received chest irradiation of 40-50 Gy. Of the 89 patients who could be wholly evaluated, the overall response rate was 78% for PE and 74% for PEI therapy (NS). For all patients the complete response (CR) rates were 14 versus 21%, respectively, and 22 versus 30% for LD. However, the median survival times for all patients were 55 weeks for PE therapy versus 56 weeks for PEI therapy (NS). The 2-year survival rates were 15 and 17%, respectively, for all patients (NS). There was no difference in the duration of response between PE and PEI therapy in cases with CR or partial response. However, severe leukopenia (< 2,000/mm3) occurred more often after PEI (73%) than after PE (44%) therapy (p < 0.05). These results suggest that PEI is not superior to PE chemotherapy in SCLC. The use of ifosfamide in multimodality treatment regimens needs to be studied further.
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Topical administration of interferon for locally recurrent renal cell carcinoma in 2 cases was reported. Case 1: a 56-year-old woman had undergone radical nephrectomy for left renal cell carcinoma. She had a locally recurrent tumor adjacent to spleen 26 months after the surgery. She received 3 million units of interferon alpha every day intralesionally for almost 3 years without any distant metastasis. Case 2: a 59-year-old man had undergone left radical nephrectomy for renal cell carcinoma. He had recurrent tumor contiguous to spleen 24 months after nephrectomy. With Seldinger method a catheter was indwelled selectively in splenic artery. Two million units of interferon gamma was administered through the catheter, twice a week for 4 weeks, and once a week in subsequent course. The tumor showed necrosis on CT film. He died of DIC 5 months after the initiation of intraarterial administration of interferon gamma. Topical use of interferon showed some favorable effects in both cases even after the failure of systemic administration. Further investigation is warranted to demonstrate the advantage of topical use of interferon over systemic administration.
Here we report a rare case of simple hepatic cysts causing dilatation of the intrahepatic bile duct. A 75-year old female was admitted to Kyushu University Hospital because of liver dysfunction. Ultrasonogram and CT scan showed two neighboring cysts in the liver (S2, S4), and dilatation of the peripheral bile duct in left lateral segment of the liver. Echo-guided percutaneous aspiration of cyst fluid showed the elevated levels of CA19-9, but malignant cell were not seen. These findings suggested that the compression by these neighboring simple hepatic cysts caused the stenosis of the bile duct. The dilatation of the bile duct disappeared after the percutaneous ethanol sclerosing therapy for cyst.
We evaluated the prognostic significance of nuclear DNA content by flow cytometry and ras oncogene expression in paraffin-embedded sections of tumors obtained surgically from 112 non-small cell lung cancer patients. Sixty-five (77%) of the 84 tumors had DNA aneuploid patterns that were statistically higher in adenocarcinoma than in squamous cell carcinoma. Of the 91 patients analyzed immunohistochemically using anti-ras Mr 21,000 protein (p21) monoclonal antibody rp-35, positive reactions (weak and strong) were observed in 56% of squamous cell carcinomas and 68% of adenocarcinomas. A better 5-yr survival rate was observed in the DNA diploid group (61%) than in the DNA aneuploid group (35%) (P less than 0.01). Patients with p21-negative tumors survived significantly longer (5-yr survival rate of 64%) than did those with p21-weak tumors (38%, P less than 0.05) or those with p21-strong tumors (12%, P less than 0.01). Cox's multivariate analysis showed that DNA ploidy, ras p21 expression, and the stage of the disease were significant prognostic factors for survival. However, the DNA content was not a major independent prognostic factor in adenocarcinoma. The intensity of ras p21 expression was not correlated with nuclear DNA content. These results suggest that DNA content or enhanced ras p21 expression may be different biological markers indicating the malignant potential of lung tumors.
Induction of carbonic anhydrase isozyme I (CA-I) by erythropoietin or hemin was investigated using erythroleukemia (K562) cells. Immunological estimation and purification of carbonic anhydrases showed that untreated K562 cells contained only carbonic anhydrase isozyme II(CA-II), while incubation of the cells with 2 units of erythropoietin (EP) per ml of the incubation medium or with 50 microM hemin resulted in the induction of CA-I. The purified CA-I induced in K562 cells was enzymatically and immunologically identical to that from mature erythrocytes. Flow cytometric analysis showed that incubation of K562 cells with EP as well as hemin induced CA-I at the 3rd h, while alpha-globin was detected at the 8th h. Northern blot analysis of CA-I mRNA using a cloned genomic DNA as a probe showed that mRNA of CA-I was induced by EP. These results suggest that induction of CA-I is regulated at the transcriptional level during developmental changes of erythroid cells, and that CA-I may play a physiologically important role during erythroid differentiation.
We investigated whether the proliferating cell nuclear antigen (PCNA) protein takes part in cis-diamminedichloroplatinum (II) (CDDP) resistance, using a murine leukemia cell line P388 and its CDDP resistant cell line. P388/CDDP was 4 times more resistant to CDDP than P388. The cell lines were maintained in the DBA/2 female mouse peritoneal space. In total cells, the amount of the PCNA protein decreased to 90% in P388 after 1 h CDDP treatment, but that of P388/CDDP increased to 127%. The difference was statistically significant (p = 0.012, n = 5). As for G2/M phase cells, the difference was also significant at 1 h (p = 0.016, n = 5) and at 2 h (p = 0.036, n = 5) after treatment. In P388 the amount of the PCNA protein decreased in accordance with the inhibited cell proliferation, whereas in P388/CDDP, the amount of the PCNA protein increased in spite of the inhibited cell proliferation. This increase of the PCNA protein suggests that the PCNA protein is involved in CDDP resistance of P388/CDDP through enhanced DNA repair synthesis.
The relationship between the quantity of silver-binding nucleolar organizer regions (AgNORs), nuclear DNA content, and proliferative activity was studied in 61 patients with adenocarcinoma of the lung. The proliferative activity of adenocarcinoma was estimated by tumor volume doubling time based on chest X-ray findings. There was a high, inverse correlation between the AgNORs and the tumor doubling time (p less than 0.001, r = -0.815), and the contribution rate was high value (2 = 0.664). However, the AgNORs value was an independent prognostic factor for survival time. A better 5-year survival rate was observed in patients with DNA diploidy than in DNA aneuploidy, but there was no statistical difference between the two groups. There was an inverse correlation between the DNA index and tumor doubling time (p less than 0.05, r = -0.565), but the contribution rate had a low value (r2 = 0.319). These results indicate that the AgNORs value is important in providing an estimate of the proliferative activity of adenocarcinoma.
To evaluate the relationship between DNA ploidy and the clinical stage of the disease or the prognosis, tumor DNA content was determined by flow cytometry in paraffin-embedded specimens obtained surgically from the patients with primary lung cancer. A good correlation in DNA indices between fresh and paraffin-embedded specimens was observed (r = 0.978), although one tumor with DNA near-diploidy was mistaken as DNA diploid in paraffin-embedded specimens. DNA content analysis was carried out for 171 of 178 patients (401 of 427 paraffin-embedded specimens). Of the 171 patients, 155 could be enrolled in the clinical and pathological study, and 118 in the prognostic study. One hundred and nineteen (76.8%) patients had DNA aneuploid patterns that were higher in adenocarcinoma than in squamous cell carcinoma (p less than 0.05) Although the proportion of DNA aneuploidy was not associated with the pathologic stage, a better 5-year survival rate was observed in Group A (DNA diploidy, 62.9%) than in Group B (DNA aneuploidy and DNA peridiploidy, 28.4%; p less than 0.001). Therefore, the DNA content was considered to be an independent prognostic factor for survival in patients with non-small cell lung cancer. To evaluate the relationship of DNA content with histopathologic heterogeneity, the morphologic study was carried out on hematoxylin and eosin stained sections in adenocarcinoma of the lung. Seventeen patients had DNA multiploidy and 10 of 17 tumors with DNA multiploidy showed prominent pleomorphism in histologic morphology. DNA content seemed to be related to histopathologic differences in non-small cell lung cancer. The measurement of DNA content is useful for evaluation of clinical behavior and prognosis of the patients with non-small cell lung cancer.
A patient with inflammatory pseudotumor of the liver associated with acute myelomonocytic leukemia (M4) is reported. He had spiking fever, epigastralgia, and elevated levels of serum C-reactive protein (CRP) and alkaline phosphatase (ALP). Ultrasonography showed a hypoechoic mass in the liver, and ultrasonically guided fine needle aspiration biopsy of the mass revealed that it was composed of fibrous connective tissue infiltrated with plasma cells, eosinophils, and neutrophils. Accordingly, a diagnosis of inflammatory pseudotumor of the liver was made. Marked reduction in the size of the lesion and a decrease of the levels of the CRP and ALP occurred without specific treatment. We emphasize the importance of ultrasonically guided aspiration biopsy in diagnosis of inflammatory pseudotumor of the liver without the need for surgery.
A simple and rapid enzyme-linked immunoassay for human erythrocyte carbonic anhydrase isozyme I was developed. The assay was found to be sensitive enough for the detection of nanogram amounts of the enzyme in incubation mixtures. The first incubation with anti-human carbonic anhydrase I IgG was carried out for 6 hours at room temperature. The second incubation of the enzyme was carried out in the presence of goat plasma C1q coupled with peroxidase for 1 h at room temperature. The enzymatic reaction was performed for 30 min using 2,2'-azino-di(3-ethyl-benzthiazoline sulfonate) as a substrate, and absorbance at 414 nm was recorded. Carbonic anhydrase I was assayed on the range of 1 to 200 ng/ml using this method. The levels of carbonic anhydrase I in K562 cells induced by erythropoietin and in other blood cells were determined. This enzyme-linked immunoassay has application for the study of developing erythroid cells.