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Biomedical subjects

Y Kawarada

Publications and source records attributed to Y Kawarada.

At least 109 records · Page 6Linked to original sources
PubMed1996

Enhancement of TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method using mung bean nuclease, a single-stranded DNA digestion enzyme.

The TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method has been employed widely to demonstrate apoptotic cells in routinely prepared paraffin sections. Because the apoptotic cells were reactive with the antibody to single-stranded DNA, we attempted to enhance the TUNEL positivity by pretreatment with single-stranded DNA digestion enzymes, S1 nuclease, and mung bean nuclease. When mung bean nuclease (5 U/50 microliter/section) was incubated at 37 degrees C for 30 min, the TUNEL reaction was most effectively enhanced. Pretreatment with S1 nuclease (0.25 U/50 microliter/section) at 37 degrees C for 45 min was less reliable. Compared with the conventional TUNEL sequence, the enhancement technique using mung bean nuclease enabled us to detect more apoptotic cells in human small intestine, colon, tonsil, thymus, endometrium, ovary, liver, kidney, and pancreas. The positivity was not affected by autolytic change. The mechanism of enhancement is discussed.

Apoptosis↗
PubMed1996

Induction and involvement of endogenous IGF-I in pancreas regeneration after partial pancreatectomy in the dog.

To elucidate whether and how IGF-I is involved in the regeneration of the pancreas after partial pancreatectomy, IGF-I mRNA expression, IGF-I protein synthesis, ornithine decarboxylase (ODC) activity and DNA replication in the remnant pancreas were determined in the dog. After pancreatectomy, IGF-I mRNA expression was remarkably enhanced in the remnant pancreas, showing the maximal value at post-operative day (POD) 1. Subsequently, IGF-I synthesis in the tissue was significantly stimulated at POD 2, and its maximal concentration was observed at POD 3. Following IGF-I synthesis, ODC activity was induced and its maximal activity was also obtained at POD 3. Finally, DNA replication was induced in the remnant pancreas, and its maximal level was observed at POD 5. These responses in the remnant pancreatic tissue to partial pancreatectomy were greatly enhanced as the resection rate was increased up to 95%. Positive correlations were observed between IGF-I concentrations in the remnant pancreas and the activities of ODC and DNA synthesis in the tissue after 95% pancreatectomy. These results suggest that the gene expression of IGF-I is rapidly induced in the remnant pancreas after partial pancreatectomy, and subsequently synthesized endogenous IGF-I peptides may stimulate ODC and other cell growth-related activities in the tissue in paracrine and/or autocrine manners eventually to induce DNA replication and tissue regeneration.

Animals↗
PubMed1996

[Current status of treatment for small hepatocellular carcinoma].

In recent years ethanol injection therapy (PEI) and transcatheter hepatic arterial embolization (TAE) have come to be widely used in the treatment of small hepatocellular carcinoma, and the introduction of microwave coagulation therapy (MCT) more recently has made it possible to perform a variety of non-surgical treatments even in cases in which surgical resection has been indicated until now. There have also been reports based on survival rates that results comparable to those obtained by surgical resection can be achieved with non-surgical methods. The main issue is whether PEI or resection should be selected to treat small hepatocellular carcinomas. However, the recurrence rate after PEI is higher than after surgical resection, and according to our results, in patients with solitary lesions, especially when the tumor diameter is 2 cm or less, the level of malignancy in many cases is also low biologically, and postoperative survival rates (recurrence-free survival rates) are favorable [5 years : 85.0% (64.3%); 10 years: 67.9% (42.2%)]. After thoroughly evaluating liver function in these cases, while surgical resection should be considered first, it is also important to use a combination of various treatment methods rather than always resort to a single method.

Carcinoma, Hepatocellular↗
PubMed1995

Copper incorporation into ceruloplasmin in rat livers.

Ceruloplasmin, a blue copper oxidase found in plasma, is synthesized in hepatocytes as a single polypeptide chain consisting of a 19 amino acid leader peptide plus 1046 amino acids of mature protein (132 kDa). Holoceruloplasmin is secreted into the plasma with 6-7 atoms of copper bound per molecule. In this study we identified apo- and holoceruloplasmin and examined the mechanism of copper incorporation during ceruloplasmin biosynthesis using the Long-Evans Cinnamon (LEC) rat which does not incorporate copper into newly synthesized ceruloplasmin. We followed the conversion from ceruloplasmin precursor (with little or no carbohydrate) to the larger product (after carbohydrate addition), which occurred in the secretory compartments of hepatocytes, by native gel electrophoresis. We found that copper accumulates in the hepatocellular Golgi apparatus of LEC rats due to a disorder in the process of copper incorporation. The data indicate that copper is incorporated into ceruloplasmin late in the course of its transport through the secretory compartments.

Animals↗
PubMed1995

The preoperative nutritional assessment of surgical patients with hepatic dysfunction.

The precise influence of malnutrition on postoperative complications in patients with hepatic dysfunction is not well known. In phase I of the present study, we evaluated the nutritional status of 102 patients with hepatic dysfunction who were admitted for elective hepatobiliary or pancreatic surgery, and a model for the prediction of postoperative complications was developed using a computer-based stepwise regression procedure. The equation for this Prognostic Nutritional Index for Surgery (PNIS) was calculated by [-0.147 x (% weight change) + 0.046 x (% ideal body weight) + 0.010 x (actual triceps skinfold thickness as a % of standard value) + 0.051 x (hepaplastintest)]. In phase II this model was prospectively tested in 182 patients, including 145 with hepatic dysfunction. A total of 18 patients were classified as having a poor prognosis (PNIS < 5) and all of these patients in fact developed postoperative complications: 128 patients were classified as having an intermediate prognosis (5 < or = PNIS < 10), 23 (18.0%) of whom developed postoperative complications, and none of the 36 patients who were classified as having a good prognosis (PNIS > or = 10) developed any postoperative complications. These results demonstrate the importance of performing a thorough preoperative nutritional assessment of patients with hepatic dysfunction as malnourished patients with PNIS < 10 may need preoperative nutritional management, even when their surgical procedures are not major.

Humans↗
PubMed1995

Role of activation of ornithine decarboxylase and DNA synthesis on ethynylestradiol-induced hepatocarcinogenesis.

We have established an animal model for estrogen-induced hepatocarcinogenesis by oral administration of synthetic hormone to female Wistar rats. Daily treatment of rats with 0.15 mg of ethynylestradiol (EE) for 4 months resulted in the development of hyperplastic foci in all animals. At 12 months of EE treatment, four of the 13 rats (30.8%) developed hepatocellular carcinoma. Ornithine decarboxylase (ODC) activity and DNA synthesis in the liver were activated by a single administration of EE, reaching peak levels at 12 and 48 h respectively. The EE-activated peak levels of both ODC activity and DNA synthesis were markedly elevated at 2 months after consecutive treatment, indicating that the female sex hormone stimulates the cellular proliferation during the initiation phase of carcinogenesis. The activities then gradually decreased, but with the levels higher than those in the controls. On the other hand, simultaneous treatment of rats with tamoxifen completely suppressed the EE-induced ODC activity, hepatic foci formation and hepatocellular carcinoma development. Together with our previous findings of DNA adducts and 8-hydroxydeoxyguanosine formation in the early stage of EE-induced carcinogenesis resulting in DNA damage, the present results suggest that estrogen enhanced ODC activity which was followed by increased DNA synthesis (DNA replication). Moreover, these effects might increase misreading during damaged DNA replication and were closely related to initiation, promotion and progression of hepatocarcinogenesis in this experimental model.

Animals↗
PubMed1995

Role of reactive oxygen in synthetic estrogen induction of hepatocellular carcinomas in rats and preventive effect of vitamins.

We have established an experimental model of oral contraceptive-induced hepatocellular carcinomas (HCCs) in female Wistar rats, revealing that ethynylestradiol (EE) and norethindrone acetate have actions as both initiators and promoters. The present time-sequence study was undertaken to clarify the role of free radicals in estrogen induction of HCC by measuring detoxifying enzyme activities and levels of 8-hydroxydeoxyguanosine (8-OH-dG) and by assessing the effects of concomitant vitamin C, vitamin E or beta-carotene administration on hepatocarcinogenesis. During 12 months oral administration of EE (0.075 or 0.75 mg/day), the 8-OH-dG levels reached peak values after 1 month, when they were significantly elevated as compared with the controls. Glutathione peroxidase demonstrated a tendency to decrease. Histologically, pre-neoplastic lesions assessed by immunohistochemical staining for placental glutathione S-transferase (GST-P) were first observed at 2 months in the groups given 0.075 and 0.75 mg/day of EE alone, with incidences of HCC at 12 months being 8.7% and 38.5% respectively. Combined administration of vitamins with 0.075 mg EE/day reduced the elevation of the 8-OH-dG levels. GST-P-positive lesions were first observed at 4 months in the vitamin E group and at 6 months in the vitamin C and beta-carotene groups. As compared with the value in the 0.075 mg EE alone group, vitamin administration significantly reduced the numbers of GST-P-positive foci after 12 months of treatment. The incidences of HCC at 12 months were 0% in the vitamin C group, 4.5% in the vitamin E group and 4.8% in the beta-carotene group, i.e. administration of the vitamins inhibited the development of GST-P-positive foci, with suppression of HCC. The results thus suggest that free radicals play an important role in the induction of HCC by estrogen.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1994

Apoptotic degeneration in the arhinencephalic brain of the mouse mutant Pdn/Pdn.

The homozygotes of a mouse strain with genetic polydactyly (Polydactyly Nagoya, Pdn) exhibit arhinencephaly and various brain malformations. In the present experiment, abnormal apoptotic degeneration in the arhinencephalic brain of Pdn/Pdn embryos and newborns was investigated immunohistochemically and by molecular genetic techniques. Polyclonal antibody against single-stranded DNA detected the nuclei of programmed dying cells (apoptotic cells) specifically in the interdigital necrotic zone of the normal mouse limb plate on day 14 of gestation. We used this antibody against single-stranded DNA to investigate the apoptotic degeneration in Pdn/Pdn brain. Abnormal apoptosis was observed in the infralimbic cortical plate, hypothalamus and periventricular thalamus on day 0 after birth in Pdn/Pdn brains. The TRPM-2 gene, which has been considered to mediate apoptosis, was expressed in the developing normal and Pdn/Pdn brains. TRPM-2 gene expressions in the brain stem and cerebellum of arhinencephalic Pdn/Pdn fetuses and newborns were higher than those of +/+ littermates. From these facts, it was suggested that the abnormal apoptosis caused a large amount of cell loss in the arhinencephalic mouse brain, and this cell loss induced the expansion of the ventricle, followed by the hydrocephaly.

Animals↗
PubMed1994

Antibody against single-stranded DNA detects both programmed cell death and drug-induced apoptosis.

Cyclophosphamide induced fragmented nuclei in mouse thymic epithelial cells. Agarose gel electrophoresis showed the fragmentation of the DNA extracted from mouse thymus exposed to cyclophosphamide. The cell death induced by cyclophosphamide was considered to be apoptotic. Polyclonal antibody against single-stranded DNA was used immunohistochemically to detect apoptotic cell death in thymic epithelial cells. This antibody also detected programmed cell death in the interdigital necrotic zone of the mouse limb plate on day 14 of gestation, and in the ganglion of the trigeminal nerve on day 13 of gestation. These results show that the antibody specific for single-stranded DNA detected both drug-induced apoptosis and programmed cell death during embryogenesis.

Animals↗
PubMed1994

Surgical treatment of hepatocellular carcinoma.

The effect of surgical treatment for hepatocellular carcinoma (HCC) was evaluated in 149 resected cases, 83.2% of which were associated with liver cirrhosis. The 3- and 5-year survival rates were 60.1% and 39.4%, respectively. The mortality rate was 4.1%. In patients aged over 70 years, liver cirrhosis was found in 53.3% of cases and the mortality rate was 6.7%. The 3- and 5-year survival rates were 50.8% and 33.9%, respectively. Factors that significantly affected survival for more than 5 years were a tumor size of less than 3 cm, Stage I disease, vp(-), IMo, and diploid type. The 5-year survival rate for patients with a single tumor of 3 cm or less was 54.2%, regardless of the surgical procedure. All 15 patients with a solitary tumor of 2 cm or less (Stage I, small liver tumor) were alive with a 5-year survival rate of 100%. The problem is the treatment of patients with a tumor measuring 3-5 cm in diameter and associated liver cirrhosis, because their prognosis after surgery is the worst. HrS (subsegmentectomy) is the minimal procedure of limited hepatectomy for these cases with postoperative multidisciplinary therapy.

Adult↗
PubMed1994

Prospective and randomized clinical trial for the treatment of hepatocellular carcinoma--a comparison between L-TAE with farmorubicin and L-TAE with adriamycin: preliminary results (second cooperative study). Cooperative Study Group for Liver Cancer Treatment of Japan.

A randomized controlled clinical trial was conducted to compare the use of farmorubicin (FARM) and adriamycin (ADR) in Lipiodol transcatheter arterial chemoembolization (L-TAE) as a treatment of hepatocellular carcinoma. In all, 192 hospitals participated, and 415 patients were enrolled in the study during the period from October 1989 through December 1990, and their data were collected. The patients were randomly allocated to group A (FARM) or group B (ADR) by a central telephone registration. Several clinical characteristics were slightly worse in group A than in group B, but there was no statistically significant difference. The actual doses of FARM and ADR were 72 mg/body and 48 mg/body, respectively. Additional treatments, including repeated TAE or surgery, were given to 248 patients. The 1- and 2-year survival rates were 69% and 44% for group A and 74% and 57% for group B, respectively. The difference was marginally significant (P value in the log-rank test, 0.038). When each group of patients was classified into two subgroups, i.e., high-risk and low-risk categories, based on the severity index calculated by the Cox regression model from significant prognostic factors, the ADR subgroup was significantly superior to the FARM subgroup in the low-risk category, but there was no significant difference between the subgroups in the high-risk category. The change in the serum alpha-fetoprotein level, the extent of Lipiodol accumulation in the tumor, and the extent of tumor reduction did not show any significant difference between the groups. At the above-mentioned doses, ADR seemed to have efficacy almost the same as or slightly superior to that of FARM in L-TAE for the treatment of hepatocellular carcinoma.

Adult↗
PubMed1994

[A case of pulmonary alveolar proteinosis with increased IgE and CEA].

We report here a case of pulmonary alveolar proteinosis with significantly increased IgE and CEA values, in both serum and bronchoalveolar lavage fluid. A 35-year-old man presented with exertional dyspnea, diffuse abnormal shadows on radiological examination of the chest and physiologic evidence of a restrictive lung disease. Bronchoalveolar lavage and transbronchial lung biopsy proved that the patient was suffering from pulmonary alveolar proteinosis. He had worked as a landscape gardener for the past 2 years, and had been exposed to insecticides. IgE and CEA levels were significantly elevated in both serum and bronchoalveolar lavage fluid. After whole lung lavage, general status improved remarkably and serum CEA fell to the normal range. The serum IgE level was, however, reduced by only half. Proliferating type II epithelial cells in pulmonary alveolar proteinosis have been suggested to produce CEA. However, the cause of the increased IgE level remains unknown. This case report suggests that some abnormal immunological events, leading to hyperproduction of IgE, could play a role in the onset of pulmonary alveolar proteinosis.

Adult↗
PubMed1992

Prospective and randomized clinical trial for the treatment of hepatocellular carcinoma--a comparison of lipiodol-transcatheter arterial embolization with and without adriamycin (first cooperative study). The Cooperative Study Group for Liver Cancer Treatment of Japan.

A randomized, controlled clinical trial comparing the use of lipiodol-transcatheter arterial embolization (L-TAE) in the presence versus the absence of Adriamycin (ADR) for the treatment of hepatocellular carcinoma was conducted from August 1988 through September 1989. In all, 125 Japanese hospitals participated in this study and 289 patients were entered in the trial. The patients were randomly allocated into group A (L-TAE) or group B (L-TAE + ADR) by telephone registration. There was no significant difference in background factors between group A and group B. Additional treatment, including repeated TAE or hepatic resection, was given to 189 patients. Among the four endpoints analyzed, the rate of tumor reduction and lipiodol accumulation in the tumor did not significantly differ between the two groups. The 3-year survival values for groups A and B were 33.6% and 34.9%, respectively; the difference was not significant. The serum alpha-fetoprotein level, however, decreased to a significantly greater extent in the group that received ADR than in the group that did not (P < 0.05). This result suggests that ADR has some favorable additional effect in L-TAE for the treatment of hepatocellular carcinoma.

Adult↗
PubMed1992

Significance of multidisciplinary therapy for hepatocellular carcinoma.

The effect of multidisciplinary therapy for hepatocellular carcinoma (HCC) was evaluated in 121 resected cases. The 5-year survival was 100% for absolute curative resection (12 cases), 59.1% for relative curative resection (n = 37) and 10.9% for relative non-curative resection (n = 59). However, none of the patients survived for more than 3 years after absolute non-curative resection (n = 13). The non-recurrence in the preoperative TAE groups was different from that in non-TAE groups undergoing absolute and relative curative resection. The 1- and 3-year non-recurrence rates for relative non-curative resection were 92.3% and 53.8%, respectively, for the preoperative TAE group and 56.1% and 28.1%, respectively for the non-TAE group. These data show that preoperative TAE is effective in relative non-curative resection. Functional disturbances of the coagulation-fibrinolysis system in cirrhotic patients were improved after PSE. All patients undergoing hepatectomy after PSE had an uneventful postoperative course, including well-maintained function of the coagulation-fibrinolysis system and a decrease in splenic volume. At 1 year after hepatectomy, cirrhotic patients with critical liver function and poor coagulation-fibrinolysis showed appreciable hepatic regeneration. One patient died of hepatic failure 1 year after the operation. In recurrent HCC, the 1-, 2- and 3-year survival values after reresection were 100%, 75.0% and 25.0%, respectively. The respective values following TAE were 79.0%, 42.0% and 9.0%. Three cases of recurrent HCC were effectively treated, i.e., two patients achieved a partial response and one showed no change, by continuous intra-arterial infusion of 5-FU and lentinan with intermittent one-shot injections of epirubicin using a subcutaneous infusion pump. These three patients are alive at 1 year and 7 months, 1 year and 4 months and 6 months after the treatment, respectively.

Adult↗
PubMed1992

Prospective and randomized clinical trial for the treatment of hepatocellular carcinoma--a comparison of L-TAE with Farmorubicin and L-TAE with adriamycin (second cooperative study). The Cooperative Study Group for Liver Cancer Treatment of Japan.

A randomized clinical trial comparing L-TAE with Farmorubicin (FARM) and L-TAE with Adriamycin (ADR) in the treatment of hepatocellular carcinoma was conducted from October 1989 through December 1990. In all, 192 hospitals participated in this study and 117 patients were entered. The patients were randomly allocated to group A (L-TAE+FARM) or group B (L-TAE+ADR). There was no significant intergroup difference in background factors. Additional treatment consisting of repeated TAE or surgery was given to 66 patients. Four factors were analyzed in this study: the percentage of reduction in tumor size, the change in the AFP level, lipiodol accumulation, and survival. None of these factors differed significantly between the two groups. The final evaluation of this study will be based on differences in survival after a long-term follow-up. Toxic effects manifested less frequently in group A than in group B, and the decrease in the platelet count in the peripheral blood was significantly lower in group A than in group B. These results suggest that FARM exerts a more favorable effect than does ADR in the treatment of hepatocellular carcinoma.

Adult↗