Search

Many temporarily functioning proteins are generated during the replacement of nucleoproteins in the nuclei of late spermatids and seem to be degraded in the nucleus. This study was designed to clarify the involvement of the ubiquitin-proteasome degradation system in the nucleus of rat developing spermatids. Thus, we studied the nuclear distribution of polyubiquitinated proteins (pUP) and proteasome in spermiogenic cells and sperm using postembedding immunoelectron microscopy. We divided the nuclear area of late spermatids into two regions: (1) a dense area composed of condensed chromatin and (2) a nuclear pocket in the neck region. The latter was located in the caudal nuclear region and was surrounded by redundant nuclear envelope. We demonstrated the presence of pUP in the dense area and nuclear pocket, proteasome in the nuclear pocket, and clear spots in the dense area of rat spermatids. Using quantitative analysis of immunogold labeling, we found that fluctuation of pUP and proteasome levels in late spermatogenesis was mostly synchronized with disappearance of histones and transitional proteins reported previously. In the nuclei of human sperm, pUP was detected in the dense area, whereas proteasome was in the nuclear vacuoles and clear spots. These results strongly suggest that pUP occur in the dense nuclear area of developing spermatids and that the ubiquitin-proteasome system is more actively operational in the nuclear pocket than dense area. Thus, the nuclear pocket might be the degradation site for temporarily functioning proteins generating during condensation of chromatin in late spermatids.

Rehabilitation of patients who have undergone bilateral maxillectomy is difficult because of extensive loss of bone and soft tissue. In this clinical report, prosthodontic rehabilitation of oral function in a bilateral maxillecitomy patient combined with a new fibular osteocutaneous flap, which was designed to have two oronasal slits for the retention of an obturator prosthesis, was described. A 58-year-old man with a maxillary alveolar carcinoma underwent bilateral maxillectomy. The defect was reconstructed using a vascularized fibular bone wrapped circumferentially with a peroneal flap, which was fixed with miniplates between the right malar prominence and cut edge of the left zygoma remaining two slits anterior and posterior to the graft. Two and half weeks after the surgery, a delayed surgical obturator was delivered and an obturator prosthesis was delivered 6 weeks after the surgery. This obturator prosthesis could be extended into the slits to engage the tissue undercuts, and was stable during use. Mastication, deglutition, articulation and the mid-facial profile of the patient were rehabilitated. After installation of the obturator prosthesis, relining of the prosthesis base was carried out alongside the healing process of the graft, and adjustment of occlusions and high-pressure spots was carried out. No clinical disorders were observed either in the grafted tissue or the obturator prosthesis with a 3-year prognosis. Newly designing a fibular osteocutaneous flap combined with tissue-borne obturator prosthesis is one successful approach to the restoration of oral function, and increases the patient's quality of life after bilateral maxillectomy.

We studied temporal changes in the subcellular localization and levels of a moonlighting protein, phospholipid hydroperoxide glutathione peroxidase (PHGPx), in spermatogenic cells and mature sperm of the rat by immunofluorescence and immunoelectron microscopy. The PHGPx signals were detected in chromatoid bodies, clear nucleoplasm, mitochondria-associated material, mitochondrial aggregates, granulated bodies, and vesicles in residual bodies in addition to mitochondria, nuclei, and acrosomes as previously reported. Within mitochondria, PHGPx moved from the matrix to the outermost membrane region in step 19 spermatid, suggesting that this spatiotemporal change is synchronized with the functional change of PHGPx in mitochondria. In the nucleus, PHGPx was associated with electron-lucent spots and with the nuclear envelope, and PHGPx in the latter region increased after step 16. In early pachytene spermatids, PHGPx signals were noted in the nuclear material exhibiting a very similar density to chromatoid bodies and in the intermitochondrial cement, supporting the previous proposal that chromatoid bodies originate from the nucleus and intermitochondrial cement. The presence of PHGPx in such various compartments suggested versatile roles for this protein in spermatogenesis. Quantitative immunoelectron microscopic analysis also revealed dynamic changes in the labeling density of PHGPx in different subcellular compartments as follows: 1). Total cellular PHGPx rapidly increased after step 5 and reached a maximum at step 18; 2). mitochondrial labeling density increased after step 1 and achieved a maximum in steps 15-17; 3). nuclear labeling density suddenly increased in steps 12-14 to a maximum; 4). in cytoplasmic matrix, the density remained low in all steps; and 5). the labeling density in chromatoid bodies gradually decreased from pachytene spermatocytes to spermatids at step 18. These spatiotemporal changes in the level of PHGPx during the differentiation of spermatogenic cells to sperm infer that PHGPx plays a diverse and important biological role in spermatogenesis.

The aim of this study was to elucidate the mechanism of the formation of the widespread mucous-plugging observed in autopsied lungs from patients with bronchial asthma. We performed morphometric analysis of airways of autopsied lungs from eight patients with bronchial asthma (Group BA), and compared it with those of six chronic bronchitics (Group CB) and four control patients (Control). The following parameters were measured in paraffin sections: volume proportion of bronchial glands to bronchial wall (Gland%); goblet cell granules to total epithelial layer (Goblet %); intraluminal mucus expressed as the mucus occupying ratio (MOR); volume ratio of intraluminal mucus continuous with goblet cells to total intraluminal mucus (Vc/Vtol %); and surface ratio of the contact surface of intraluminal mucus continuous with goblet cells to the total luminal surface (Sc/Stot %). Gland%, Goblet %, and MOR or inflammatory cell numbers in the airway walls both from Group BA and CB were larger than those from the Control group. However, no significant differences were observed between Group BA and CB in Gland%, Goblet %, MOR or inflammatory cell numbers, except for the eosinophil number: i.e. 23 +/- 3, 22 +/- 3 and 6 +/- 2% in Gland%; 22 +/- 9, 5 +/- 4 and 2 +/- 2% in Goblet%; 10 +/- 3, 18 +/- 3 and 0.3 +/- 0.5% in MOR; 199 +/- 68, 10 +/- 3 and 2 +/- 2 cells. mm-2 in eosinophil number of the peripheral airways from Groups BA, CB and Control, respectively. In contrast, marked and significant increases were observed both in Vc/Vtot% and Sc/Stot% in Group BA compared to Groups CB and Control both in central and peripheral airways: i.e. Vc/Vtot% in the peripheral airways was 53 +/- 5, 4 +/- 3 and 0.8 +/- 0.8% from Groups BA, CB and Control, respectively (BA vs CB or BA vs Control, p < 0.01 each). These findings suggest that the continuity of goblet cells and intraluminal mucus or lack of full release of mucus, from goblet cells, is peculiar to asthmatic airways, and may contribute to the formation of mucous-plugs.

We report a case of small cell esophageal carcinoma (SCEC) with multiple liver metastases treated with some success by chemotherapy with irinotecan (CPT-11) plus cisplatin (CDDP). Radiologic and endoscopic examination of a 75-year-old man with multiple liver tumors disclosed a 4.0-cm type 2 tumor in the middle third of the esophagus. An endoscopically obtained biopsy specimen was diagnosed as undifferentiated small cell carcinoma. Multiple liver metastases were confirmed but lymph node metastases and distant metastases other than those in the liver were not detected. After six courses of chemotherapy with CPT-11 plus CDDP, the primary lesion showed complete response and liver metastases showed partial response. However, because all lesions almost immediately relapsed or progressed, arterial infusion chemotherapy for liver metastases and radiation for the primary lesion were given as second-line treatment. The primary lesion showed complete response with radiation. Arterial infusion chemotherapy prevented the progression of liver metastases once, but the patient died of liver failure at last. No distant lesions including metastatic lymph nodes were confirmed over the course of his illness, and the patient survived for a year after first diagnosis. Although the prognosis of SCEC is quite unfavorable due to highly aggressive behavior, a better prognosis is possible with effective chemotherapy and second-line treatment is important in improving prognosis.

This preliminary investigation, involving 422 patients, tested the hypothesis that rate of eating is associated with obesity in patients with type 2 diabetes or hyperlipidaemia at all ages. The patients' eating habits were determined using a questionnaire, and the patients were classified as quick, normal or slow eaters. The body mass indices of the three groups were compared. The body mass indices of the male patients who ate quickly (25.4 ± 0.2 kg/m2) were significantly higher than those of the patients who ate at a normal rate (24.4 ± 0.3 kg/m2) or slowly (24.1 ± 0.5 kg/m2). No difference between body mass indices in the female groups was found. It was speculated that rate of eating affects body weight in male patients with type 2 diabetes or hyperlipidaemia.

The validity of mass screening using fecal occult blood testing remains controversial. In addition, no controlled clinical study has yet been performed to show the usefulness of sigmoidoscopy. The purpose of the present study was to compare the surgical results achieved in asymptomatic patients with colorectal cancer detected by screening with those in symptomatic individuals.A total of 285 patients underwent a surgical resection of colorectal cancer between 1991 and 1997 at our institution. Among them, 233 patients had complaints related to cancer at the time of diagnosis. In contrast, 52 were asymptomatic. In those 52 patients, colorectal cancer had been suspected based on routine screening including fecal occult blood testing, colonoscopy and/or elevated serum levels of carcinoembryonic antigen.Early stage of colorectal cancer was more frequently seen in asymptomatic patients than in symptomatic patients P0.01. The survival rates for asymptomatic patients was also superior to those of symptomatic patients P0.05.Screening using fecal occult blood testing, colonoscopy and tumor markers is thus considered to be beneficial for the early detection of colorectal carcinoma, which also tends to demonstrate good surgical results.

A 72-year-old woman was admitted because of anorexia and dyspnea. She was given a diagnosis of pulmonary hemorrhage and renal failure. Despite treatment with high-dose steroid and hemodialysis, the patient died of disseminated intravascular coagulation on the 9th hospital day. Autopsy revealed intra-alveolar hemorrhage, crescentic glomerulonephritis, and systemic vasculitis with fibrinoid necrosis. A direct immunofluorescence study demonstrated linear deposition of IgG along the glomerular basement membrane (GBM). Both anti-GBM antibody and anti-neutrophil cytoplasmic antibody with perinuclear pattern (P-ANCA) were detected in the patient's serum by enzyme immunoassay. Goodpasture's syndrome with P-ANCA was diagnosed. There has been some controversy as to whether vasculitis occurs in patients with Goodpasture's syndrome. This was a rare example of well-documented Goodpasture's syndrome with P-ANCA and systemic vasculitis, the exact etiologic relationships among which remain to be clarified.

It is important to detect genes that may be good prognostic markers for colon cancer patients. With this in mind, we identified the motility related protein-1 (MRP1/CD9) gene in human colon tissues. The aim of this study was to clarify the significance of MRP1/CD9 gene expression in human colon cancers. We performed the differential mRNA display technique between tumor/normal paired samples of the colon and identified MRP1/CD9. Eighty-two surgical specimens of primary colorectal cancer were analyzed by means of reverse transcription-PCR for the MRP1/CD9 gene. Its expression status and clinicopathological variables were analyzed univariately and multivariately. The MRP1/ CD9 mRNA expression was positive in 56 cases and negative in 26 cases. The MRP1/CD9 negative cases showed a significantly higher frequency of venous-vessel invasion and liver metastasis, or a worse prognosis than the MRP1/CD9 positive cases (P0.05). Multivariate analysis with the Cox regression model disclosed that MRP1/CD9 expression was an independent prognostic factor distinct from the lymph node status. The findings imply that the study of MRP1/CD9 expression may be useful for predicting prognosis of patients with colorectal cancer.

The 12 members of the human melanoma antigen-encoding (MAGE) gene family encode tumor-specific peptide antigens. Some antigens coded by the MAGE genes are potentially useful for cancer-specific immunotherapy. However, little information on the expression of these genes in human colon carcinomas is available. We investigated the expression of 10 of the 12 genes in human colon tissue.Eighty pairs of tumor and normal tissue samples from the human colon were studied by means of reverse transcription-polymerase chain reaction.None of the genes was expressed in the 80 control samples of normal tissue. On the other hand, expression was recognized in tumor samples, ranging from 5% of samples for MAGE-6 to 44% for MAGE-8. Seventy of the 80 tumor samples (88%) expressed at least 1 of the 10 MAGE genes. The frequency of liver metastasis was significantly higher in cases with tumor samples that expressed MAGE-3 than in those that did not express this gene. This tendency was not observed for other members of the MAGE gene family. No significant differences were observed in the other clinicopathologic factors between any MAGE-positive and -negative tumor cases.The MAGE genes were exclusively expressed in carcinoma tissues and not in normal tissues of the colon. The finding that nearly 90% of tumors expressed at least one MAGE gene indicates the possible clinical use of this gene for both immunotherapy and molecular diagnosis.

MAGE genes code tumor antigens that are recognized by cytolytic T lymphocytes and have been shown to be expressed in various malignant tumors. However, there is still little information on the expression of the MAGE gene family except for reports of MAGE-1 and -3. In this study, we therefore investigated the expression of MAGE-4, -6, -8, -9, -10, -11 and -12, as well as MAGE-1, -2 and -3 in both cell lines and surgical samples of gastric carcinoma, using reverse transcriptionPCR. Of the investigated 11 cell lines, MAGE-4, -6, -8, -9, -10, -11 and -12 were detected in 8 (73%), 6 (55%), 2 (18%), 59 (44%), 6 (55%), 4 (36%), and 7 (64%), respectively. No expression of these genes was seen in any of the 54 samples of normal gastric tissue. In contrast, the tumor tissue samples were found to express MAGE-4, -6, -8, -9, -10, -11, and -12 in 18 (33%), 13 (24%), 6 (11%), 10 (19%), 5 (9%), 13 (24%), and 10 (19%), respectively. Forty-four (82%) of 54 gastric tumors expressed at least one of these genes. No significant correlation was observed between the expression of MAGE genes and any specific clinicopathological factors. These results may hopefully prove to be useful in developing strategies for tumor-specific immunotherapy of gastric carcinoma using MAGE gene products.

Three agents, verapamil, cepharanthine, and 2-[4-(diphenylmethyl)-1-piperazinyl]ethyl-5-(trans-4,6-dimethyl-1, 3,2-dioxaphosphorinan-2-yl)-2,6-dimethyl-4-(3-nitrophenyl)-3-py ridinecarboxylate P-oxide (PAK-104P), that reverse drug resistance in P-glycoprotein (P-Gp)-mediated multidrug-resistant cells were examined for their activity to reverse drug resistance in multidrug resistance-associated protein (MRP)-mediated multidrug-resistant C-A120 cells. Agents other than PAK-104P could not reverse the resistance to doxorubicin in C-A120 cells. PAK-104P moderately reversed the doxorubicin resistance. In contrast, PAK-104P almost completely reversed the resistance to vincristine (VCR) in C-A120 cells as well as in KB-8-5 cells, and other agents moderately reversed the VCR resistance in C-A120 cells. PAK-104P at 10 microM enhanced the accumulation of VCR in C-A120 cells to the level of that in KB-3-1 cells without the agent. PAK-104P competitively inhibited the ATP-dependent [3H]leukotriene C4 uptake in membrane vesicles isolated from C-A120 cells. These findings demonstrate that PAK-104P can completely reverse the resistance to VCR in both P-Gp- and MRP-mediated multidrug-resistant cells and that PAK-104P directly interacts with MRP and inhibits the transporting activity of MRP.

We compared cardiac output values obtained by the following 3 methods: 1. Iced bolus injectate method, using the injectate temperature actually measured for computation [bolus cardiac output with flow-through temperature probe: BCO (p)]. 2. Continuous cardiac output measurement, using Vigilance system (continuous cardiac output: CCO). 3. Iced bolus injectate method, assuming injectate temperature to be 0 degrees C (bolus cardiac output without flow-through temperature probe: BCO). Only the subject receiving cardiovascular surgeries using cardiopulmonary bypass (CPB), and requiring a thermodilution catheter for hemodynamic monitoring during anesthesia were selected for the study. The total cases consisted of 15 patients (10 males), in which 60 measurement were made. Following the anesthetic induction with fentanyl, midazolam, vecuronium bromide, and intubation, a Baxter Swan-Ganz CCO thermodilution catheter was inserted through the right internal jugular vein. Compared to BCO(p), CCO showed excellent precision and bias; (r2 = 0.935, P0.0001, bias = -0.19 l.min-1, SD = 0.36 l.min-1). Although BCO correlated well with BCO(p), the bias was much greater (r2 = 0.919, P0.0001, bias = -1.25 l.min-1, SD = 0.45 l.min-1) than BCO(p). In conclusion, the values obtained by BCO(p) and CCO methods were very accurate. On the other hand, a considerable overestimation was found with BCO since the compensation for actual injectate temperature was not performed

To improve the surgical outcome in patients with radiation enteritis, 18 female cases were analyzed. Out of the 18 cases, 3 (16.7%) were treated conservatively while 15 (83.3%) underwent surgical procedures. Thirteen out of 14 patients (92.8%) with ileus underwent an operation. The overall mortality was 22.2% (4 out of 18 cases). However, no significant difference in the mortality between the operated and non-operated cases was observed. Although an analysis of the 15 operated cases did not reveal any significant factors that might have affected the prognosis, all four patients who underwent a bypass operation showed a good postoperative course, with only one excepting being a patient suffering from malnutrition. Although only a small number of patients were included in this study, these results suggest that 1) surgeons should not hesitate to operate on patients with radiation enteritis demonstrating ileus, and 2) a bypass operation may be one surgical alternative in the presence of massive adhesion or for patients at high risk for a standard operation.

It has been reported that several genes may be good indicators for determining biological behavior, including the prognosis, of colorectal cancers. We have summarized these reported genes, such as tumor suppressor gene, oncogenes, metastasis suppressor gene, adhesion molecules, growth factors, proteinases, and others, including microsatellite instability. Some of the genes such as p53, DCC, c-met, or matrix metalloproteinase are considered to be reliable for determining biological aggressiveness. We introduced several interesting genes which we are focusing using cDNA subtraction library analysis. We hope that these genes are well combined for best analysis of the biological behavior of colorectal cancers and use for practical clinical analysis. In addition, we hope that novel important genes indicative for prognosis will be found.

Intraoperative transesophagial echocardiography (TEE) was performed in a 27-year-old man who underwent direct suture for atrial septal defect (ASD) under general anesthesia. Myocardial ischemia occurred about five minutes after weaning from cardiopulmonary bypass, and at that time TEE showed that anteroseptal wall, lateral septal wall and lateral wall are severely hypokinetic and another wall is diffusely hypokinetic in the left ventricle after detecting air bubbles in the left ventricle by TEE. The changes in TEE preceded the ST elevation in the ECG. Bolus infusion of isosorbide dinitrate and continuous infusion of nitroglycerin showed no effects on these changes. Ten minutes after the onset of myocardial ischemia, these changes were alleviated. TEE enabled us to detect air bubbles and evaluates myocardial ischemia before the appearance of ST changes in ECG.

Expression of ganglioside alpha-2,8-sialyltransferase (GD3 synthetase; EC 2.4.99.8) gene and gangliosides in human leukemia cells were analyzed. Flow cytometric analysis of various types of leukemia cells using monoclonal antibodies revealed that GD3 was expressed at a moderate level on adult T-cell leukemia (ATL) cells and weakly on T-cell acute lymphocytic leukemia cells. Accordingly, high levels of GD3 synthase mRNA in ATL and low levels in T-cell acute lymphocytic leukemia were demonstrated by semiquantitative reverse transcription-PCR analysis. High-level expression of the GD3 synthase gene in ATL cells was detected in the uncultured state, and was not changed during short-term culture in vitro. This was in contrast with the rapid up-regulation of GM2/GD2 synthase gene, which was observed within 24 h after start of culture after the emergence of human T-lymphotropic virus type I (HTLV-I) gene expression in cultured normal T lymphocytes with or without HTLV-I p40tax protein, GD3 synthase gene was almost equally expressed, whereas GM2/GD2 synthase gene was much more strongly expressed in the p40tax-positive T cells. These findings indicated that the GD3 synthase gene was highly expressed in ATL cells unrelated to HTLV-I genome expression.

Intraoperative transesophageal echocardiography (TEE) was performed on a 62-year-old man who underwent abdominal aortic replacement for abdominal aortic aneurysm under general anesthesia combined with epidural anesthesia. Coronary artery spasm occurred after unexpected massive hemorrhage, and TEE showed hypokinesis in the posterior-inferior left ventricular wall. The changes in TEE preceded the ST elevation in the ECG. Bolus infusion of isosorbide dinitrate and continuous infusion of nitroglycerin alleviated these changes. TEE enabled us to detect and evaluate coronary spasm before the appearance of ST changes in ECG.

Human thymidine phosphorylase (dThdPase) has been reported to be identical with the platelet-derived endothelial cell growth factor (PD-ECGF). To investigate whether the dThdPase activity of PD-ECGF/dThdPase is indispensable to its angiogenic activity, three PD-ECGF/dThdPase mutants, K115E (Lys-115--Glu), L148R (Leu-148--Arg) and R202S (Arg-202--Ser) were made by site-directed mutagenesis. Although the expression level of the three mutant PD-ECGF/dThdPases in the COS-7 cells was similar to that of wild-type PD-ECGF/dThdPase, dThdPase activity was not detected in the COS-7 cells transfected with the mutant PD-ECGF/dThdPase cDNA. The lysates of COS-7 cells transfected with the wild-type PD-ECGF/dThdPase cDNA had angiogenic activity, but those transfected with the mutant PD-ECGF/dThdPase cDNAs did not. An inhibitor of dThdPase, 6-amino-5-chlorouracil, inhibited the angiogenic activity of purified dThdPase. These findings indicate that dThdPase activity is indispensable to the angiogenic activity of PD-ECGF/dThdPase.

The substrate specificity of beta 1,4-N-acetylgalactosaminyltransferase has been analyzed using a fusion enzyme which consisted of the catalytic domain of the enzyme and the IgG binding domain of protein A, and also by extracts from cDNA transfectants. Both enzyme sources were capable of producing not only GM2 and GD2, but also asialo-GM2, GalNAc-sialylparagloboside, and Gal-NAc-GD1a from appropriate acceptors, although the efficiencies were at most 1-3% of those of GM2/GD2. The biological significance of these low specificities was studied with transient and stable transfectant cells. From the results of transient expression of the cDNA, asialo-GM2 expression appeared to inversely correlate with GM2 synthase levels in those lines. Consequently, GM2 seemed to be preferentially synthesized when both GM3 and lactosylceramide are available, and asialo-GM2 is synthesized in the absence of GM3 synthesis. However, the results of double immunostaining of CHO transfectants with anti-GM2 and anti-asialo-GM2 antibodies indicated that another factor may be involved in asialo-GM2 synthesis. From the in vitro assay using mixed acceptors, it was concluded that the presence of certain levels of GM2 might enhance the asialo-GM2 synthesis. These results suggest that even acceptors showing low efficiencies in vitro might be used in certain cells depending on the availability of precursors, expression levels of other gangliosides, as well as the kinetic properties of the enzyme, and the compartmentation of the glycosylation machineries in the cells.

For the isolation of ganglioside GD3 synthase (EC 2.4.99.8) cDNA, we developed an expression cloning approach that used an anti-GD2 monoclonal antibody for selection. A host recipient cell line that we have named KF3027-Hyg5 was also utilized. This cell line expresses high levels of GM2 as well as GM3 but no GD3 or GD2 and was constructed from mouse B16 melanoma cells transfected with the polyoma large tumor antigen gene (KF3027) and the previously cloned beta-1,4-N-acetylgalactosaminyltransferase (EC 2.4.1.92) cDNA. Four rounds of transfection, monoclonal antibody 3F8 panning, and Hirt extraction resulted in the isolation of two cDNA clones, transfection of which directed the expression of GD3 in KF3027 and B16 melanoma cells and GD3 and GD2 in KF3027-Hyg5 cells. The cDNA contained a 1650-bp insert and a single open reading frame. The deduced amino acid predicted a type II membrane topology consisting of cytoplasmic (14 aa), transmembrane (18 aa), and catalytic (309 aa) domains. The sequence also predicted the presence of a sialyl motif similar to that found in the other sialyltransferases cloned so far. As expected, mRNA of this gene (2.6 kb) was strongly expressed in human melanoma lines.

Recently, it has been recognized that there are increasing incidences of hepatocellular carcinoma (HCC) multicentricity. Thus, intraoperatively detected hepatic lesions that were once thought to be metastatic lesions now need to be carefully reexamined to determine whether they are true metastatic lesions or the multicentric development of HCC.We investigated the histological characteristics of small nodular lesions detected during intraoperative ultrasonography in 33 consecutive patients with small HCC who underwent laparotomy at our institution.Fourteen nodular lesions were found incidentally in 10 of 33 patients (30.3%), and were classified into the following three groups: 11 nodules in nine patients (27.3%) were HCC, two nodules in two patients (6.1%) were hemangioma, and one nodule in one patient (3.0%) was a large regenerative nodule. HCC therefore comprised 78.6% of the intraoperatively detected nodular lesions. Of the 11 HCCs, six were hyperechoic, four were hypoechoic, and one was isoechoic. Five (83.3%) of six small hyperechoic HCCs and two (50.0%) of four hypoechoic HCCs were well differentiated and retained their preexisting liver structure. These findings closely coincide with the characteristics of early stage HCC. Thus, early stage HCC comprised 63.6% of the intraoperatively detected HCC cases.A certain proportion of small satellite HCCs detected during intraoperative ultrasonography in patients with small HCC, which were previously thought to be metastatic lesions from the main HCC, may instead be early stage HCCs. Such findings would also support the concept of the multicentric development of HCC. Approximately 60% of all small HCC cases detected intraoperatively may be early stage HCC. As a result, it is predicted that the emergence of HCC is either multicentric or unicentric, with early intrahepatic spread, although the former seems to be more common.

We report a case of pneumonitis due to Rikkunshi-to. A 79-year-old woman was admitted to our hospital because of interstitial pneumonia. She complained of dry cough and dyspnea on exertion. Fine crepitations were heard on physical examination. The chest X-ray film revealed diffuse reticulonodular shadows in both lung fields. Under the suspicion of drug-induced pneumonitis, all drugs were stopped and she was given prednisolone. Consequently her complaints, laboratory data and chest X-ray findings markedly improved. Microscopic examination of transbronchial lung biopsy specimens showed interstitial pneumonia. The results of lymphocyte stimulation test and lymphocyte migration inhibition test were positive for Rikkunshi-to. Based on these findings, we diagnosed this case as pneumonitis due to Rikkunshi-to. To our knowledge, there has been no previous case of pulmonary hypersensitivity due to Rikkunshi-to reported in the world.

We report a case of early gastric cancer with Virchow's node metastasis. The patient underwent partial gastrectomy and postoperative immunochemotherapy using MMC, 5'-DFUR and PSK, which reduced the Virchow's node. Three years after surgery, we found metastases to the left subclavicular and axillary nodes other than the Virchow's node. Then UFT was administered orally at 600 mg/day, and the metastatic nodes diminished, then vanished. The patient is alive nearly five years after surgery.

Thymidine phosphorylase (dThdPase) is an enzyme involved in pyrimidine nucleoside metabolism. However, little is known about its physiological functions. We previously purified dThdPase from human placenta, isolated a complementary DNA clone for this enzyme, and sequenced it. There was complete sequence identity between 120 amino acids of human dThdPase and the sequence of platelet-derived endothelial cell growth factor (PD-ECGF). Human KB epidermal carcinoma cells transfected with platelet-derived endothelial cell growth factor complementary DNA expressed a 55-kDa protein that was detected with anti-dThdPase antibody and the cell lysate had dThdPase activity. The sensitivity of transfected cells to the antimetabolites was compared with that of untransfected KB cells. The sensitivity of the transfected cells to Doxifluridine (5'-deoxy-5-fluorouridine) was higher than that of untransfected KB cells. Transfected cells were also more sensitive to Tegafur than untransfected KB cells. These results demonstrate that dThdPase is involved in the activation of these anticancer agents. Since many cancer tissues contain high dThdPase activity compared with normal tissues, these transfected and untransfected KB cells are useful for studying the role of dThdPase in the activation of pyrimidine antimetabolites and also in angiogenesis.

We describe a new model of human ischemia encephalopathy in rabbits by means of cross-clamping the ascending aorta in combination with blood exsanguination from the right atrium. Rabbit hippocampal CAI pyramidal cells and cerebellar Purkinje cells showed only a few changes in the early period after ischemia, but 4 days after ischemia a significant decrease in cell count and marked degeneration of the remaining cells were observed. When the pathological findings 4 days after TCI in the hippocampal area and cerebellar area were compared, the rate of cytoclasis was found to be significantly higher in the latter than in the former.

We resected a multiple hepatocellular carcinoma (HCC) which contained portal tracts, showed evidence of an early multistep condition, and a multicentric development of HCC which progressed simultaneously. A preoperative working examination revealed a tumor in S5 of the liver, which was 2.5 cm in diameter. In addition, a "nodule-in-nodule" appearance was seen on the ultrasonographic imaging. At intraoperative ultrasonography, an additional two HCCs were detected. At histologic examination of the resected specimen, the main nodule showed a nodule-in-nodule appearance. In the outer nodule, there was a well-differentiated HCC, while the portal tracts remained. The less-differentiated HCC grew in the inner nodule and was replacing the well-differentiated HCC in the outer nodule. These findings suggests morphological transition in the early stages of the multistep development of HCC. Two additional tumors (1.1 cm and 1.0 cm in diameter) detected at intraoperative ultrasonography proved to be HCCs (grade I) with marked fatty change, and with portal tracts remaining within both the HCCs. Furthermore, both of the HCCs retained their preexisting liver structure. These histologic findings coincided with the characteristics of the early stage HCC, whereas the coexistence of early stage HCCs suggests the multicentric development of HCC. Distortion, compression, and invasion of the portal tracts may appear as the tumor grows, thereby evoking an increasing risk of metastasis via the portal tracts. Therefore, the early diagnosis and treatment of HCC before the portal tracts disappear, when 1.5 cm in size or less, may be very important for surgeons and diagnosticians, as an effective curative resection may be feasible.

Partial complementary DNA (cDNA) for thymidine phosphorylase (dThdPase) was cloned by means of a polymerase chain reaction. There was complete sequence identity between the amino acid sequence deduced from the nucleotide sequence of a clone (288 nucleotides) and the residues of platelet-derived endothelial cell growth factor (PD-ECGF). The amino acid sequence of all four peptide fragments from purified human dThdPase could be aligned with that of PD-ECGF. Our data indicate that residues 125-244 of PD-ECGF are identical to the sequence of human dThdPase. The molecular weights of human dThdPase and recombinant PD-ECGF (rPD-ECGF) that lacks 10 amino acids at the amino terminal were 55 and 52 kDa, respectively. Anti-PD-ECGF antibody recognized dThdPase, and anti-dThdPase antibody recognized rPD-ECGF. rPD-ECGF had dThdPase activity and its specific activity was similar to that of purified human dThdPase. dThdPase activity and molecules were detected in COS cells transfected with human PD-ECGF cDNA, but not in nontransfected cells. The sizes of PD-ECGF and dThdPase in the transfected COS cells were identical. These data suggest that human dThdPase is identical to PD-ECGF.

Between 1965 and 1985, 194 of 1,113 patients (17.4 percent) with advanced carcinoma of the stomach who underwent gastric resection had Borrmann type IV carcinoma of the stomach, a macroscopically evident and diffusely spreading tumor. These patients tended to be younger and female and to have larger tumors that involved the entire stomach, as compared with patients with other types of carcinomas. Tissues were commonly undifferentiated, serosal invasion was prominent with infiltrative growth and high rates of metastasis to the lymph nodes and peritoneal dissemination were evident. In most instances, the disease was advanced; therefore, total gastrectomy was performed upon 82.0 percent of the patients and a palliative resection was done in 64.4 percent. The survival rate was lower than for patients with carcinoma of the stomach of other types (p less than 0.01). A multivariate analysis indicated that curative resection is an independent favorable prognostic event, while other factors are dependent covariates. Early detection of this carcinoma is crucial to extend survival time of patients with Borrmann type IV carcinoma of the stomach. Aggressive postoperative chemotherapy should be considered when a noncurative resection is done.

From 1965 to 1985, 1,150 patients underwent gastrectomy for carcinoma of the stomach invading beyond the submucosa. One thousand, one hundred and forty-one patients were studied for five years or longer. The patients were classified into two groups--those treated from 1965 to 1974 (n = 622) and those treated from 1975 to 1985 (n = 519). In the more recent group, there was a significant increase in the number of patients more than 70 years of age, in the number of female patients, of carcinomas present in the upper two-thirds of the stomach, in tumors exceeding 10 centimeters and diffusely infiltrative type and in early cancer-simulating type of advanced carcinoma in gross appearance and in undifferentiated type adenocarcinomas in histology. Although differences in the incidence of patients treated by radical procedures, such as extensive lymph node dissection and combined resection of adjacent organs, were not statistically significant, the incidence for those who underwent total gastrectomy was significantly increased. The over-all long term survival rate remained unchanged during 1965 to 1985 (five year survival rate of 31.3 versus 32.8 per cent). However, there was a significantly longer survival period for patients who underwent total gastrectomy (five year survival rate of 22.9 versus 28.3 per cent) (p less than 0.05). Much of the improvement can be attributed to an increase in the detection of relatively small advanced carcinomas and of advanced carcinomas of the stomach simulating early carcinoma in the upper one-third of the stomach. Therefore, we emphasize the need to diagnose advanced carcinoma of the stomach when the lesion is in an earlier stage of disease.

From 1965 to 1985, of 1108 patients with advanced gastric cancer who underwent gastric resection in our department, 216 patients (19.5%) had synchronous peritoneal dissemination (PD) or liver metastasis (LM). The 1-year survival rate was 22.5% for PD, 14.7% for LM, and 4.8% for PD plus LM (p less than 0.01). With PD, patients were younger, tumor was larger, Borrmann type 3 and 4 and undifferentiated lesions were more frequent, and the incidence of serosal invasion was higher. Histologic findings showed a pattern of infiltrative growth. With LM, patients were more likely to be male, Borrmann type 2 and 3 and differentiated lesions were frequent, and the antral region was often involved. The degree of serosal invasion was less than with PD. The pattern of growth was the expansive type and vascular involvement was prominent. Rate of lymph node metastasis was high in both groups. The route of metastasis was partly linked to features of the primary lesion. Multivariate analysis showed that independent risk factors involved in the occurrence of each metastasis are serosal invasion, lymph node metastasis, and undifferentiated tissue type for PD, and lymph node metastasis and vascular involvement for LM. When designing treatment in an attempt to suppress a recurrence, even after curative resection, all of these risk factors must be kept in mind.

DNA ploidy was determined by cytofluorometric analysis of paraffin-embedded malignant tissue from 96 Japanese patients in whom gastric carcinoma had invaded the serosa. Aneuploidy was found in gastric carcinoma tissue from 63 patients (66%). The postoperative 5-year survival rate of patients with aneuploid malignancy was significantly lower (13%) than those with diploid malignancy (36%) (p less than 0.05). A multivariate analysis of various clinical and pathologic factors showed that tumor size, lymph node metastasis, vascular invasion, and DNA ploidy were significant and independent factors, which correlated with prognosis.

From 1965 to 1985, 356 of 1,468 patients (24.3 per cent) with carcinoma of the stomach who underwent gastric resection had disease arising from the upper one-third of the stomach. Tumors in the upper one-third of the stomach were larger, and gross appearance of types 3 and 4 was frequent. Serosal invasion was prominent, and the rates of metastases of the lymph nodes and liver were higher in carcinoma in the upper one-third of the stomach compared with carcinoma in other regions of the stomach. Palliative resection was done for 43.8 per cent of the patients. Survival rate for patients with carcinoma in the upper one-third of the stomach was lower than for patients with lesions in other regions of the stomach (p less than 0.01). Multivariate analysis indicated that operative curability, hepatic metastasis, serosal invasion, lymph node metastasis, peritoneal dissemination and tumor size are significant prognostic factors for carcinoma in the upper one-third of the stomach. These events, except for peritoneal dissemination, differed between carcinoma in the upper one-third of the stomach and in other regions of the stomach, and the survival rate for patients with carcinoma of the upper one-third of the stomach was less favorable. Early detection is crucial for improving the survival time of patients with carcinoma in the upper one-third of the stomach. Aggressive postoperative chemotherapy should be considered when noncurative resection is done.

Proliferative activities in 91 primary gastric carcinomas and 36 corresponding metastatic perigastric lymph nodes were investigated using Ki-67 labeling percentage and an argyrophilic nucleolar organizer region (AgNOR) count. Tumors with a high proliferative activity often metastasized to lymph nodes, and the proliferative activities of the primary lesion and the perigastric lymph node metastases were similar. A significant correlation was recognized between the Ki-67 labeling percentage and the AgNOR count (r = 0.744; P less than 0.001). The Ki-67 labeling percentage and AgNOR count proved to be useful predictors of nodal metastasis regardless of tumor size, depth of invasion, and histological type. Even when tumors are smaller (less than 7 cm) or the stage of the disease is early (pT1, 2), the formation of metastasis increased with an increased Ki-67 labeling percentage or AgNOR count. The combination analysis of depth of invasion with Ki-67 labeling percentage or AgNOR count gives a more precise prediction of nodal metastasis, compared with histological analysis alone.

To assess the reliability of the exercise ECG in detecting silent ischemia, ECG results were compared with those of stress-redistribution thallium-201 single-photon emission computed tomography (SPECT) in 116 patients with prior myocardial infarction and in 20 normal subjects used as a control. The LV was divided into 20 segmental images, which were scored blindly on a 5-point scale. The redistribution score was defined as thallium defect score of exercise subtracted by that of redistribution image and was used as a measure of amount of ischemic but viable myocardium. The upper limit of normal redistribution score (= 4.32) was defined as mean +2 standard deviations derived from 20 normal subjects. Of 116 patients, 47 had the redistribution score above the normal range. Twenty-five (53%) of the 47 patients showed positive ECG response. Fourteen (20%) of the 69 patients, who had the normal redistribution score, showed positive ECG response. Thus, the ECG response had a sensitivity of 53% and a specificity of 80% in detecting transient ischemia. Furthermore, the 116 patients were subdivided into 4 groups according to the presence or absence of chest pain and ECG change during exercise. Fourteen patients showed both chest pain and ECG change and all these patients had the redistribution score above the normal range. Twenty-five patients showed ECG change without chest pain and 11 (44%) of the 25 patients had the abnormal redistribution. Three (43%) of 7 patients who showed chest pain without ECG change had the abnormal redistribution score. Of 70 patients who had neither chest pain nor ECG change, 19 (27%) had the redistribution score above the normal range. Thus, limitations exist in detecting silent ischemia by ECG in patients with a prior myocardial infarction, because the ECG response to the exercise test may have a low degree of sensitivity and a high degree of false positive and false negative results in detecting silent ischemia.

Two hundred and seventy-seven patients with advanced carcinoma of the stomach invading the serosa were studied with regard to the relationship between the length of the serosal invasion (LSI) and prognosis, with a division made between expanding and infiltrative types of tumors. Among patients with expanding types of tumors, five year survival rates were 40 per cent in patients with LSI less than 4.0 centimeters, 33 per cent in patients with LSI 4.1 to 6.0 centimeters and 13.0 per cent in patients with LSI more than 6.0 centimeters. Among those patients with infiltrative types of tumors, those with less than 2.0 centimeters of LSI had a better prognosis (five year survival rate of 53 per cent), whereas those patients with a LSI more than 2.0 centimeters had a very poor prognosis (five year survival rate of less than 20 per cent) and the significant LSI on prognosis was found to be 2.0 centimeters. Involvement of the lymph node and metastasis to the liver were not affected by LSI, whereas the rates of peritoneal dissemination and direct invasion to neighboring organs increased with an increase in LSI. Knowledge of these factors facilitates the planning of postoperative treatment.

The purpose of this prospective randomized study was to determine whether prophylactic administration of urinary trypsin inhibitor prevents postoperative damage to the pancreas caused by R2 gastrectomy in patients with gastric cancer, by analyzing related enzyme activities. Among the 22 patients who underwent distal partial gastrectomy together with R2 lymphadenectomy, 12 were given the drug for 3 days postoperatively, and 10 no therapeutic agent. These groups were otherwise comparable. Postoperatively, the control patients had significantly higher levels of total amylase activity in the serum and P-type amylase when compared to the preoperative data. These amylase activities almost remained at the preoperative level in those given the drug. When the ratio of increase in enzyme activities (postoperative value divided by preoperative value) were compared, on the 7th postoperative day, total amylase activity in the serum and P-type amylase were significantly different between the two groups (p less than 0.05). This difference was also evident when comparing total amylase activity in the urine (p less than 0.05). These findings indicate that prophylactic administration of urinary trypsin inhibitor would aid in preventing postoperative hyperamylasemia caused by R2 gastrectomy in patients with gastric cancer.

A 72-year-old female was admitted because of a palpable hard tumor, 15 X 7 cm in size, in the left lower quadrant of the abdomen. According to a diagnosis of retroperitoneal tumor, laparotomy was performed, revealing a large tumor extending between the descending colon and the pelvic cavity, which could not be separated from the retroperitoneum, uterus, left ovary, urinary bladder or rectum. A number of metastatic nodules were disseminated to the peritoneum and mesenterium. The pathological diagnosis of the nodules was poorly differentiated carcinoma of transitional cell type, origin unknown. After discharge, oral administration of bestrabucil (100-200 mg/day), the benzoate of an estradiol-chlorambucil conjugate, was given to the patient at an outpatient clinic. The tumor ceased to be palpable when the total amount administered reached at 14 g. Ultrasonography and computed tomography demonstrated significant reduction in tumor size. A decrease in the peripheral leukocyte count was observed after every consecutive administration and it took about 2 months to recover to the normal level. No other side effects were observed in this case. Although administration was stopped at a total amount of 16.1 g, reenlargement of the tumor has not been observed for 9 months.

The correlation between the growth pattern and peptic ulceration occurring in the cancer lesion was studied in early gastric cancer. Although the superficially spreading type (Super), which grows slowly, showed a high rate of being accompanied by peptic ulceration (74.0%), the deeply penetrating type (Pen), with rapid growth, had a lower such rate (Pen A- expansive type 9.7%; Pen B- infiltrative type 16.1%). Deep ulceration accompanied 32% of the Super cases, 12% of the small mucosal cases and no cases of the Pen type. The Super and small mucosal types located at the intermediate gland area had peptic ulceration more frequently than those at the other areas. The rate of the cases of peptic ulceration of undifferentiated adenocarcinoma was high in the Super and small mucosal types, and that of differentiated adenocarcinoma was 64.0% in the Super and only 33.3% in the small mucosal type. This study clarified that the growth of early gastric cancer was inhibitively influenced by peptic ulceration, which was modified by the localization and histologic type of the carcinoma.