Search PubMed⌕ Search

Biomedical subjects

F Nakayama

Publications and source records attributed to F Nakayama.

At least 37 records · Page 2Linked to original sources

Profiling of eicosanoids in inflamed gall bladder wall by gas chromatography with selected-ion monitoring.

The profiling of eicosanoids, including prostaglandin E2 (PGE2), prostaglandin F2 alpha (PGF2 alpha), 6-keto-prostaglandin F1 alpha (6-keto-PGF1 alpha), thromboxane B2 (TXB2) and leukotriene B4 (LTB4), in dog and human gall bladders was carried out by a combination of an effective and convenient clean-up procedure and gas chromatography with selected-ion monitoring. The clean-up procedure was based on the stepwise elution of their methyl ester derivatives from a silica gel column with n-hexane-ethyl acetate and ethyl acetate-methanol in various ratios. The LTB4 methyl ester was eluted with an n-hexane-ethyl acetate (2:1, v/v) fraction because LTB4 is more lipophilic than the other eicosanoids. The present method permitted the quantitation of trace amounts of eicosanoids, including LTB4, present in tissues in the order of pg/mg of protein, without interference from other endogenous substances. In experimental acalculous cholecystitis produced in dog, the levels of eicosanoids (except LTB4) were significantly changed. Of these eicosanoids, the level of 6-keto-PGF1 alpha was significantly higher in the seromuscular layer and correlated with the observed severe morphological changes. In human chronic cholecystitis with gallstones, the level of 6-keto-PGF1 alpha in the mucosal layer was significantly higher than that in the seromuscular layer. These data suggest that prostaglandin I2 may play an important pathophysiological role in the course of cholecystitis.

6-Ketoprostaglandin F1 alpha↗

Fibronectin: a possible factor promoting cholesterol monohydrate crystallization in bile.

To examine the hypothesis that fibronectin physiologically present in bile might be a possible nucleating factor, the concentrations of fibronectin in gallbladder bile were determined and its induced effect on nucleation time and on the form of vesicle were examined in bile-model and human gallbladder bile. The gallbladder bile samples taken from patients with cholesterol gallstone had a significantly higher concentration of fibronectin and the faster nucleation time than the control. However, no significant correlation was found between nucleation time and endogenous fibronectin concentration. The addition of 0.5, 1.2, 10 micrograms/ml of fibronectin into two kinds of bile-model significantly shortened the nucleation time in a dose-related manner. Nucleation time was significantly shortened by the addition of 1 microgram/ml exogenous fibronectin into abnormal bile while such an effect was absent in the control. The addition of fibronectin increased the size of vesicles observed by the electron microscope. The results suggest that fibronectin physiologically present in bile may be one of the possible nucleating factors.

Analysis of Variance↗

Improved delineation of the gallbladder wall with ultrasonography: its value in assessment of the depth of carcinoma invasion.

We examined 30 surgically removed gallbladders with ultrasound using a 10 MHz transducer in a water bath. The sonographic structure of gallbladder wall was imaged as 5 layers. The layer 3, a well-defined hyperechoic zone, was confirmed histologically as corresponding to the boundary between subserosa and muscularis. Since the spreading of gallbladder carcinoma beyond the muscle coat is an important prognostic indicator, high resolution images may well be used to advantage in assessing gallbladder wall penetration intraoperatively, enabling the surgeon to choose the proper operative procedure.

Adenocarcinoma, Papillary↗

Promoting effect of truncal vagotomy on pancreatic carcinogenesis initiated with N-nitrosobis(2-oxopropyl)amine in Syrian golden hamsters.

The effect of truncal vagotomy (TV) on pancreatic carcinogenesis initiated with N-nitrosobis(2-oxopropyl)amine (BOP) was investigated in 81 female Syrian golden hamsters. The animals were divided into four groups according to the treatment, groups 1 and 2 serving as non-initiated controls receiving a single s.c. injection of 0.9% NaCl followed by either a sham operation or TV respectively, at week 2. Groups 3 and 4 were given a single s.c. injection of 70 mg/kg body wt of BOP before the sham operation or TV. All hamsters were killed at week 24, and the pancreas, liver and gall bladder tissues were examined histologically. While TV itself caused no significant change in pancreatic weight, the incidence of pancreatic carcinomas in hamsters from group 4 was 48.4%, significantly higher than the 16.7% evident in hamsters from group 3 (P less than 0.05). GLC analysis of the bile acid composition of gall bladder bile from hamsters not receiving carcinogen 1 and 4 months after TV revealed significantly decreased secondary bile acids. The results thus indicated that changes in bile acid composition may be involved in enhancement of BOP-initiated pancreatic carcinogenesis in hamsters by TV.

Animals↗

Manometric and histologic assessment following proctocolectomy and straight enteroanal anastomosis in canines.

Serial manometric studies were performed in mongrel dogs before and after total colectomy, mucosal proctectomy and interposed jejunoanal anastomosis without pouch. Fecal continence and body weight gain were clinically satisfactory by 28 weeks after surgery. Even at 28 weeks after the mucosal proctectomy, the mean maximal resting anal sphincter pressure was significantly lower than the preoperative level (p less than 0.001). Neorectal compliance also decreased. The rectoanal inhibitory reflexes were not observed 2 weeks after the mucosal proctectomy but atypical to typical patterns gradually appeared up to 28 weeks following the mucosal proctectomy. Histologic examination showed no injury of anal sphincter muscles. Mild inflammation was found in the neorectal mucosal layer in all animals. Meissner's plexus in rectal muscular cuff was almost completely disrupted. These data suggested that there is certain limitation in restoration of anoneorectal function after mucosal protectomy and enteroanal anastomosis, though with gentle and careful operations.

Anal Canal↗

Increased lysophosphatidylcholine and pancreatic enzyme content in bile of patients with anomalous pancreaticobiliary ductal junction.

A high incidence of inflammation and carcinoma of the biliary tract in patients with anomalous pancreaticobiliary ductal junction has been well documented. The change in biliary phospholipids as a result of the reflux of pancreatic juice into the biliary tract through anomalous pancreaticobiliary ductal junction may be responsible for it. We developed a new method of analysis of phospholipid classes using aminopropyl Bond Elut cartridge for extraction and high-performance liquid chromatography for separation. Satisfactory recovery was achieved (i.e., more than 95% for both phosphatidylcholine and lysophosphatidylcholine). With this method, the bile of 11 patients with anomalous pancreaticobiliary ductal junction was examined. The concentration and proportion of lysophosphatidylcholine in bile were much higher in the presence of anomalous pancreaticobiliary ductal junction than in controls (3.44 +/- 1.50 mmol/L vs. 0.52 +/- 0.25 mmol/L, 60.0% +/- 31.0% vs. 2.3% +/- 1.4% in gallbladder bile; p less than 0.001). In contrast, the concentration of phosphatidylcholine and the sum of phosphatidylcholine and lysophosphatidylcholine in gallbladder bile significantly decreased (p less than 0.001), but in hepatic bile they did not. An inverse correlation was found between the proportion of lysophosphatidylcholine and phospholipid concentration in gallbladder bile. Phospholipase A2 and amylase activities in bile were markedly high. Increased total fatty acid concentration and proportion of unsaturated fatty acid in bile were found. Total bile acid concentration in gallbladder bile was significantly lower than in controls. These results suggest that a considerable amount of lysophosphatidylcholine, which is known to have a cytotoxic effect, isp reduced by phospholipase A2 in refluxing pancreatic juice, and an increased concentration of lysophosphatidylcholine gives rise to cell damage causing mucosal hyperplasia and metaplasia.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

[Localization of HTLV-I-associated antigens in adult T cell leukemia cells and HTLV-I-infected cell line cells].

Localization of HTLV-I-associated antigens was studied in adult T cell leukemia (ATL) cells and HTLV-I-infected cell line cells using monoclonal and human polyclonal antibodies against the viral-related antigens. Two monoclonal antibodies that we obtained by hybridoma technique reacted with HTLV-I-virus core antigens, P19 and P24, respectively. Human anti-HTLV-I-antibodies, which were purified from sera from ATL patients reacted with not only HTLV-I virus particles but also their precursors located in the cytoplasm. In tumor cells freshly isolated from ATL patients, no expression of the virus antigens was observed. When the cells were cultured for several days, the virus antigens were defined in about 3-5% of the cultured cells by the monoclonal antibodies, and in 5-10% by the purified human anti-HTLV-I antibodies. Addition of 5-iodo-2'-deoxyuridine to the culture inhibited cell growth, and at the same time, increased the percentage of the virus antigen-positive cells. Established HTLV-I-infected cell lines showed different cytological profiles from the original ATL cells in the viral replication and morphology.

Antibodies, Monoclonal↗

Simultaneous determination of keto and non-keto bile acids in human serum by gas chromatography with selected ion monitoring.

A reliable method for the simultaneous determination of keto and non-keto bile acids in human serum was developed. Carbonyl substituents of bile acid ethyl esters were converted into methyloxime and hydroxyl substituents into dimethylethylsilyl ethers and the products were analysed directly by capillary gas chromatography with selected ion monitoring using [2H4]chenodeoxycholic and [2H4]3 alpha-hydroxy-7-oxo-5 beta-cholanoic acids as internal standards. The bile acid peaks on the selected ion chromatogram were separated without interference from endogenous substances present in serum. Recoveries of individual keto bile acids added to serum range from 74.4 to 94.7% with a mean of 87.1%. Eight kinds of keto bile acids not previously found in sera of normal subjects, namely 3-oxo-, 3-oxo-7 alpha-hydroxy-, 3-oxo-12 alpha-hydroxy-, 3 alpha-hydroxy-7-oxo, 3 alpha-hydroxy-12-oxo-, 3-oxo-7 alpha,12 alpha-dihydroxy-, 3 alpha,7 alpha-dihydroxy-12-oxo- and 3 alpha,12 alpha-dihydroxy-7-oxo-5 beta-cholanoic acids were identified and quantified. The total concentration of keto bile acids was found to be 0.16 +/- 0.08 nmol/ml and constituted 2.9 +/- 1.5% of that of the usual non-keto bile acids in peripheral venous serum.

Adult↗

Bile acid composition in brown pigment stones.

The bile acids in brown pigment stones and gallbladder bile were fractionated into free acids, glycine and taurine conjugates, and sulfates using diethylaminohydroxypropyl Sephadex LH-20 (DEAP-LH-20) column chromatography and were quantitated by gas chromatography. Twenty-eight cases of brown pigment stones were studied and divided into two groups: those with and those without bacteria possessing bile acid-deconjugating activity. In the former, free bile acid amounted to 62 +/- 34% of the total bile acid, while in the latter, only 0.1% of total bile acid was free bile acid. The fraction of total bile acid made up of free bile acids was found to be consistently higher in brown pigment stones than in the corresponding bile, irrespective of the presence or absence of biliary infection. Free bile acid is present in negligible amounts in normal bile. Total bile acid concentration in the bile of patients with brown pigment stones was significantly less than that of controls (13 vs 50 mg/ml). Biliary infection is almost always present in cases with brown pigment stones. These findings suggest that bacterial infection is present at the initiation of brown pigment stone formation as well as during the period of ensuing stone growth.

Bacterial Infections↗

Effects of bile acid feeding on hepatic deoxycholate 7 alpha-hydroxylase activity in the hamster.

In order to investigate the effects of bile acid feeding on hepatic microsomal deoxycholate 7 alpha-hydroxylase activity, three different bile acids were administered (0.2% w/w in chow) to hamsters for two weeks. Deoxycholate 7 alpha-hydroxylase activity was increased markedly by feeding of cholic acid (CA) and slightly by deoxycholic acid (DCA). Chenodeoxycholic acid (CDCA) had little effect on the enzyme activity. Feeding each of the bile acids significantly inhibited the activity of cholesterol 7 alpha-hydroxylase in the order CDCA greater than or equal to DCA greater than CA. There was no correlation between deoxycholate 7 alpha-hydroxylase activity and cholesterol 7 alpha-hydroxylase activity. It is concluded that the activity of deoxycholate 7 alpha-hydroxylase is up-regulated by feeding DCA and CA and that the mechanism seems to be different from that of cholesterol 7 alpha-hydroxylase. The increased activity of hepatic deoxycholate 7 alpha-hydroxylase by CA and DCA should be beneficial in minimizing the toxic effects of DCA in the hamster.

Alkaline Phosphatase↗

Unscheduled DNA synthesis after treatment with 20-methylcholanthrene and N-nitrosodimethylamine in primary culture of human gallbladder epithelial cells.

Unscheduled DNA synthesis (UDS) induced by treatment with 20-methylcholanthrene (MCA) and N-nitrosodimethylamine (DMN) was measured in cultured human gallbladder epithelial cells. MCA induced UDS very efficiently, while DMN was far less effective than MCA. Addition of rat S9 mixture did not affect the amount of UDS by the chemicals. Differences between the present results in human cells and our previous findings in bovine cells could be due to species and tissue specificity.

Cells, Cultured↗

Homogenous junctional sequence of the V14+ T-cell antigen receptor alpha chain expanded in unprimed mice.

Nucleotide sequences of VJ (variable-joining) junctional regions of V14+ alpha-chain T-cell receptor genes show that most V alpha 14+ T cells use one alpha chain (V alpha 14J alpha 281 with a one-nucleotide N region, which is frequently used in keyhole limpet hemocyanin-specific suppressor T-cell hybridomas) in unprimed mice. Moreover, the frequency of this alpha-chain expression was greater than 1.5% of the total alpha chains found in laboratory strains, including B10 congenic mice. This is about 10(4) times higher than was expected. The V14J281 alpha-chain expression was relatively low but was significant in CD4+/CD8+ immature thymocytes and became quite high in mature single-positive T cells, implying that this alpha chain is selected during T-cell maturation. V14J281 expression increased with time after birth and reached a maximum at around 5 weeks of age. The ligand seems to be a self molecule and to be present in laboratory strains but to be absent in a wild mouse, Mus musculus molossinus, because bone marrow chimeras clearly showed that bone marrow cells derived from Mus musculus molossinus negative for this alpha chain raised V14J281-positive T cells in a C57BL/6 environment. The above results suggest that there are some selection mechanisms for this cell type other than those for conventional alpha beta T cells and also that the homogenous VJ junction of the V14J281 alpha chain plays a pivotal role in the selection of the T cell and its ligand reactivity.

Animals↗

A peculiar form of epidermotropism in cutaneous metastatic carcinoma.

A 71-year-old man developed metastatic cutaneous nodules of colon carcinoma on the scalp and chest wall. Histopathological findings revealed aggressive invasion of the tumor cells into the overlying epidermis (epidermotropic carcinoma); they occasionally adhered to the epidermal cells. These morphological features have not been described in cutaneous metastases from internal carcinomas.

Adenocarcinoma↗

Growth kinetics of fibroblasts from a patient with Ehlers-Danlos syndrome.

A 6-year-old girl with Ehlers-Danlos syndrome (type I) who shows hyperelastic skin, hypermobile joints and delayed wound healing is reported. The growth kinetics of fibroblasts derived from her skin was examined using 3H-thymidine autoradiography. Our study showed that these fibroblasts had poor proliferative activity, compared with normal age-matched fibroblasts. Thus, we could suggest that the pathogenesis of Ehlers-Danlos syndrome was partially attributed to the lack of proliferative activity of the fibroblasts.

Autoradiography↗

Esterified and total 7 alpha-hydroxycholesterol in human serum as an indicator for hepatic bile acid synthesis.

Serum levels of 7 alpha-hydroxycholesterol and activities of hepatic microsomal cholesterol 7 alpha-hydroxylase in surgical patients were analyzed by capillary gas-liquid chromatography-selected ion monitoring technique using a new internal standard, 5 alpha-cholestane-3 beta, 7 beta-diol. We found that concentrations of 7 alpha-hydroxycholesterol obtained after alkaline hydrolysis were higher than those without alkaline hydrolysis, indicating that a considerable amount of 7 alpha-hydroxycholesterol in human serum is present in esterified form. Esterified 7 alpha-hydroxycholesterol could also be quantitatively hydrolyzed with cholesterol esterase, suggesting that fatty acid is bound at the 3 beta-position of the cholestenediol. The serum levels of esterified and free 7 alpha-hydroxycholesterol in patients with cholelithiasis were 198.0 +/- 90.3 and 48.3 +/- 19.8 pmol/ml (mean +/- SD), respectively, and were similar to those in patients without hepatobiliary diseases. After treatment with chenodeoxycholic acid (300 mg per day) for 7 to 10 days, esterified and free 7 alpha-hydroxycholesterol levels decreased to 64.9 +/- 33.6 and 20.5 +/- 11.1 pmol/ml, respectively. Activity of cholesterol 7 alpha-hydroxylase was also inhibited. Treatment with ursodeoxycholic acid (600 mg per day) for 7 to 10 days had no inhibitory effect on serum 7 alpha-hydroxycholesterol levels and the enzyme activity. In all groups, high correlations were found between the activity of cholesterol 7 alpha-hydroxylase and serum levels of 7 alpha-hydroxycholesterol: free (r = 0.71, n = 38, P less than 0.001); esterified (r = 0.87, n = 38, P less than 0.001); total (r = 0.87, n = 38, P less than 0.001). Esterified and total 7 alpha-hydroxycholesterol was more highly correlated with the enzyme activity than the free form. We conclude that a significant amount of 3 beta-acyl esters of 7 alpha-hydroxycholesterol is present in human serum and that serum levels of esterified and/or total 7 alpha-hydroxycholesterol are likely to reflect the activity of hepatic cholesterol 7 alpha-hydroxylase and thus the amount of primary bile acids synthesized in the liver.

Adult↗

[Flow cytometric cell cycle analysis of lymphocytes in patients with drug eruption].

DNA synthesis of lymphocytes was analyzed by flow cytometry in patients with drug eruption, eczema, viral eruption, and normal individuals. Lymphocytes were taken from the patients having fresh skin rash. In the patients with drug eruption, the percentages of lymphocytes in the G0G1 phase were lower and those in S + G2M phase were higher than the values in other groups, indicating the occurrence of blastogenic reaction. We considered that this method was helpful for diagnosis of drug eruption.

Adult↗