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Biomedical subjects

Y Fujiyama

Publications and source records attributed to Y Fujiyama.

142 records · Page 8Linked to original sources
PubMed1985

Clostridium ramosum, an IgA protease-producing species and its ecology in the human intestinal tract.

A bacterial strain isolated from feces of a patient with ulcerative colitis, which had been shown to produce a novel immunoglobulin A (IgA) protease (cleaving both the human IgA1 subclass and IgA2 subclass of A2m(1) allotype) extracellularly, was identified as Clostridium ramosum. By using a selective medium (propionate-rifampicin-gentamicin-colimycin-polymyxin medium) devised for C. ramosum, analysis of the population level of this organism was performed to determine its ecology in the human intestinal tract. C. ramosum was isolated in 20 of 25 fecal samples (80%) from patients with inflammatory bowel disease (I.B.D.) and in 112 of 135 samples (83%) from patients without I.B.D. (control group). C. ramosum was also isolated from 6 of 11 biopsy samples (55%) of the inflamed rectal mucosa from patients with ulcerative colitis and from five of 15 samples (33%) from the intact mucosa of the control group. The population levels of C. ramosum in most of the biopsy samples ranged from 2.3 to 5.0 log10 per gram. The IgA protease-positive C. ramosum was found in only four of 135 fecal samples (3%) and one of 15 biopsy samples (6.7%) from the control group. These results indicate that IgA protease-positive C. ramosum is not likely to play a role in the induction of I.B.D., unless the organism is first isolated from the patient with I.B.D.

Clostridium↗
PubMed1985

Metabolism of n-3 polyunsaturated fatty acids and modification of phospholipids in cultured rabbit aortic smooth muscle cells.

The metabolism of the linolenic acid family (n-3) of fatty acids, e.g., linolenic, eicosapentaenoic, and docosahexaenoic acids, in cultured smooth muscle cells from rabbit aorta was compared to the metabolism of linoleic and arachidonic acids. There was a time-dependent uptake of these fatty acids into cells for 16 hr (arachidonic greater than docosahexaenoic, linoleic, eicosapentaenoic greater than linolenic), and the acids were incorporated mainly into phospholipids and triglycerides. Eicosapentaenoic and arachidonic acids were incorporated more into phosphatidylethanolamine and phosphatidylinositol plus phosphatidylserine and less into phosphatidylcholine than linolenic and linoleic acids. Docosahexaenoic acid was incorporated into phosphatidylethanolamine more than linolenic and linoleic acids and into phosphatidylinositol plus phosphatidylserine less than eicosapentaenoic and arachidonic acids. Added linolenic acid accumulated mainly in phosphatidylcholine and did not decrease the arachidonic acid content of any phospholipid subfraction. Elongation-desaturation metabolites of linoleic acid did not accumulate. Cells treated with eicosapentaenoic acid accumulated both eicosapentaenoic and docosapentaenoic acids mainly in phosphatidylethanolamine and the arachidonic acid content was decreased. Added docosahexaenoic acid accumulated mainly in phosphatidylethanolamine and decreased the content of both arachidonic and oleic acids. The following conclusions are drawn from these results. The three n-3 fatty acids are utilized differently in phospholipids. The arachidonic acid content of phospholipids is reduced by eicosapentaenoic and docosahexaenoic acids, but not by linolenic acid. Smooth muscle cells have little or no desaturase activity, but have significant elongation activity for polyunsaturated fatty acids.

Animals↗
PubMed1985

A novel IgA protease from Clostridium sp. capable of cleaving IgA1 and IgA2 A2m(1) but not IgA2 A2m(2) allotype paraproteins.

Three bacterial strains of Bifidobacterium and Clostridium sp. from patients with inflammatory bowel disease (I.B.D.) and Streptococcus pneumoniae from a patient with pneumonia were identified to produce extracellular proteases cleaving IgA into Fab and Fc fragments. Although the proteases from the Bifidobacterium and the Streptococcus pneumoniae showed the characteristics of typical IgA1 proteases, cleaving the IgA of only the IgA1 subclass, the protease from Clostridium sp. revealed a dual substrate specificity, in that it cleaved both IgA1 and IgA2 of the A2m(1) allotype. The latter protease, however, did not show any activity with respect to the IgA2 of the A2m(2) allotype. Fc fragments isolated from the IgA1 and the IgA2 A2m(1) by digestion with the Clostridium sp. protease were identified to have an identical amino terminal residue of valine. The site of cleavage in both the alpha 1 and the alpha 2 of A2m(1) by the protease was assumed to be an identical peptide bond at Pro(221)-Val(222), which is a common one present just before the hinge of both the alpha 1 and the alpha 2 of the A2m(1) but not of the alpha 2 of the A2m(2). The protease was sensitive to ethylene-diamino tetraacetic acid, a chelating agent, similar to other already reported IgA1 proteases.

Bacterial Infections↗
PubMed1984

Immunochemical identification of an alpha 1-acid glycoprotein-antigenic determinant on carcinoembryonic antigen (CEA) and non-specific cross-reacting antigen (NCA).

An immunochemical characterization of carcinoembryonic antigen (CEA) and NCA (non-specific cross-reacting antigen) was performed. Positive reactions of CEA and NCA (Mr 60 000) with some antibodies to alpha 1-acid glycoprotein (AG) were observed. Thus, both antigens may contain immune determinants in common with alpha 1-acid glycoprotein. CEA showed positive reactivity with anti-NCA. NCA showed positive reactivity with either polyclonal or monoclonal antibodies to CEA, but negative reactivity with auto-antibodies to CEA. 125I-Tetracosapeptide (synthetic peptide-24 corresponding to the amino terminal sequence of CEA) failed to react with any antisera against CEA, NCA and AG. 125I-AG also showed no immuno-reaction with any antibody against CEA, NCA and tetracosapeptide. These results suggest that some monoclonal antibodies to CEA are directed against a common antigenic determinant of both CEA and NCA in addition to AG and tetracosapeptide, and that the auto-antibody to CEA is directed against a unique immune determinant which is not common to NCA. Thus, CEA appears to contain a unique determinant not found in NCA. Similarities in the composition of both amino acids and carbohydrates of CEA and NCA suggest that CEA is 'big-big' AG and NCA is 'big' AG.

Antibodies, Monoclonal↗
PubMed1984

Role of aging in lipid metabolism in the aorta of rats.

To clarify the intrinsic process of aging in aorta, the lipid metabolism was investigated in serum, aorta, and liver of 24-month-old rats in comparison to 10-month-old animals. Serum total cholesterol and the lipid content of low-density lipoprotein slightly increased in aged animals. Enzyme activities which hydrolyze or synthesize cholesterol ester or triglyceride in aorta were enhanced in aged animals. The increase in hydrolysis overcame that in synthesis, suggesting that enzyme activities were not changed in order to accumulate cholesterol ester and triglyceride in the aorta with aging. Phospholipid content and choline phosphotransferase activity were low both in the aorta and liver of aged animals. Moreover the phospholipid to free cholesterol ratio was decreased in aged animals, suggesting a change of membrane property with aging.

Aging↗
PubMed1984

[Cell kinetics and nuclear ploidy pattern in relation to the growth of gastric cancers as analyzed by DNA-RNA cytofluorometry].

We investigated the cell kinetics and nuclear ploidy pattern of human gastric cancers (12 early and 30 advanced cancers) using DNA-RNA cytofluorometry (NIKON SPM-RF1-D) with AO stain. The results showed that the gastric cancers studied could be divided into two main groups on the basis of ploidy pattern determined both by DNA and RNA contents: group I without, and group II with polyploidization. Cells having nuclear DNA contents between 2 n and 4 n were regarded as representing those in the S phase, and it was found that both groups had similar proliferative activity. In group I, cell proliferation without polyploidization seemed to be maintained during tumor growth regardless of the extent of invasion, and the fraction of 2 n cells was 72-94%, compared to 93-99% in control cells from the non-neoplastic gastric epithelium. In group II, however, the extent of polyploidization appeared to be increased in association with both the tumor growth and its invasion into deeper tissues; thus the fraction of 2 n cells was low, ranging from 12 to 79%. Especially, the fraction of 2 n cells in cancers with aneuploid-polyploidization was further decreased to 6-25%. Based on these results, it is hypothesized that, in the early stages of gastric cancers, the cell population may be composed mostly of diploid cells, but with both further tumor development and its invasion, the neoplastic cells may gradually differentiate into two distinct cell populations.

Adult↗
PubMed1983

Neonatal monosodium glutamate-induced lesions of hypothalamus increase intestinal fat absorption in adult mice.

The hypothalamic lesion induced by monosodium glutamate injection during the suckling period affected the absorption of fat from the intestine. Mice in group 1, in which the hypothalamic lesion included not only the arcuate nucleus, but also the basal preoptic area and two-thirds of the ventromedial nucleus, showed more efficient rates of fat absorption with high serum concentrations of immunoreactive insulin than the control. Mice in group 2, in which the hypothalamic lesion was confined mostly to the arcuate nucleus, also showed a significant increase in fat absorption, but elevated immunoreactive insulin was not found in the serum of these mice.

Aging↗
PubMed1983

Production of antibody for alpha 1-acid glycoprotein using carcinoembryonic antigen and normal fecal antigen.

The structural characteristics of carcinoembryonic antigen (CEA) and CEA-like antigen in feces and meconium were examined using antibody raised against 2 antigens. When antibody to CEA was made, anti-alpha 1-acid glycoprotein (AG) (nonprecipitating antibody) besides the precipitating antibody for CEA was made. When antibodies against each CEA-like antigen (mol. wt. 180,000) purified from feces and meconium were tested, each antibody showed a fused precipitin line for 2 antigens. Some antibodies showed the non-precipitating antibody for AG. CEA and CEA-like antigen in feces and meconium may be composed of 2 portions (AG portion and non-AG portion). Antibody specific to these antigens may be the antibody directed to the non-AG portion.

Animals↗
PubMed1982

Purification of carcinoembryonic antigen by affinity chromatography with anti-alpha 1-acid glycoprotein.

Purification of radiolabeled carcinoembryonic antigen (CEA) preparations by affinity chromatography with anti-AG bound to Sepharose was attempted, since an immunological similarity between AG (alpha 1-acid glycoprotein) and a portion of CEA had been noted. When 125I-CEA was purified in this manner, the fraction which did not bind to the column showed decreased reactivity with either anti-AG or anti-CEA. The retained fraction showed enhanced reactivity with both anti-AG and anti-CEA. The yield of purified CEA increased when the CEA preparation was allowed to react with the anti-AG column overnight. Purification of CEA from tumor tissue was performed by affinity chromatography. A perchloric acid (PCA) extract from cancer tissue was mixed with antiserum against CEA to give an immune complex, and a CEA-reactive fraction obtained by PCA extraction. The CEA-reactive fraction was eluted from a Sephadex G-200 column, and final purification was by anti-AG chromatography. When purified CEA was applied to a Sephadex G-200 column with carrier protein after labeling with 125I, the eluted radioactivity was found only in the 180,000 dalton fraction. Almost all the radioactivity was precipitated from the labeled protein by either anti-AG or anti-CEA. Purification of CEA is possible by affinity chromatography with anti-AG bound to Sepharose.

Animals↗
PubMed1982

Immunological similarity of CEA with alpha 1-acid glycoprotein (orosomucoid).

We have demonstrated that 125 I-CEA (carcinoembryonic antigen) from two commercial sources (Roche and CIS RIA kit) can be precipitated by antibody to alpha 1-acid glycoprotein (AG) dose-dependently. The binding of 125 I-CEA to anti-AG can be displaced by unlabelled AG, though, there was no cross-reaction between AG and anti-CEA. The data strongly suggest that CEA has an immunological similarity to AG. The perchloric acid extract from cancerous tissue was fractionated on a Sephadex G-200 column, and the first eluted fraction (containing large amounts of CEA) was subjected to affinity chromatography using anti-AG bound to Sepharose. The bound fraction was eluted, labelled with 125 I, and then applied to a Sephadex G-200 column with carrier protein. The radioactivity was found mainly in the large molecular size eluted fraction (Fr 1). Almost all radioactivity of Fr 1 was precipitated by anti-AG and anti-CEA. This experiment also demonstrated that a large molecular weight component (Mr: 180 000) obtained from tumor was immunologically related to both CEA and AG. Based on differences in binding affinities between CEA with anti-CEA and with anti-AG we postulate that CEA may be a large molecular precursor of AG (big form of AG).

Carcinoembryonic Antigen↗
PubMed1982

Immunological similarity of NCA (non-specific cross-reacting antigen) in feces with alpha 1-acid glycoprotein.

We have recently suggested that carcinoembryonic antigen (CEA) may contain alpha 1-acid glycoprotein (AG) antigenic determinant. In the present work we examined a protein with CEA-like activity in the feces of healthy subjects (NCA) for immunological cross-reactivity with AG. When the perchloric acid extract of feces was fractionated on a Sephadex G-200, two fractions (large and small molecular weight) were obtained. The large molecular weight fraction had higher CEA activity than the small one. The perchloric acid (PCA) extract of feces was subjected to affinity chromatography using anti-CEA bound to Sepharose, and the bound protein was labelled with 125I, and then fractionated on a Sephadex G-200 column. Two radioactive peaks, Peak 1 corresponding to an approximate Mr of 180 000 and Peak 2, corresponding to an approximate Mr of 60 000 were found. Both peaks showed immunoreactivity with either anti-CEA or anti-AG. This experiment suggests the presence of two kinds of CEA-reactive proteins in feces: one which may be a big protein with immunological similarity to AG and a second which appears to be a hydrolysed fragment of this protein.

Antigens↗
PubMed1982

Lipid metabolism in the aorta and the brain microvessels of rabbits on high cholesterol diet.

The characteristics and properties of lipid metabolism in the aorta and the brain microvessels of rabbits were investigated to clarify the role of lipid metabolism in formation of atherosclerosis. In rabbit aorta, cholesterol esterase and lipase each had an acidic and a neutral optimum pH, whereas acyl-CoA synthetase, acyl-CoA: cholesterol acyltransferase, triglyceride synthesizing activity and choline phosphotransferase each had one neutral optimum pH. These pH optima were similar to those in rats. High cholesterol diet induced atheromatous lesions in the aorta, but not in the brain microvessels. In atheromatous aorta, the acid lipase and acid phosphatase activities were higher than in controls, but not the acid cholesterol esterase activity. Moreover the activities of neutral lipase, acyl-CoA synthetase, acyl-CoA:cholesterol acyltransferase, triglyceride synthesis and choline phosphotransferase were increased, but neutral cholesterol esterase activity was normal. These data suggest that lipis metabolism in the atheromatous aorta is changed in a manner favoring accumulation of lipids, especially cholesterol esters. In controls, most of the above enzyme activities in the brain microvessels were higher than those in the aorta. However, these enzyme activities in the brain microvessels were not changed by cholesterol feeding. Thus it is suggested that the properties of lipid metabolism in the aorta and brain microvessels, including permeability of lipoproteins into the vessel walls, are important in formation of atherosclerosis in addition to the serum factors.

Animals↗
PubMedDate not supplied

Laparoscopy-guided intracorporeal ultrasound accurately delineates hepatobiliary anatomy.

UNLABELLED: The purpose of this study was to develop a technique and assess the ability of a laparoscopic ultrasound probe to delineate biliary antomy and to determine the presence or absence of duct stones. METHODS: Five pigs had ultrasonography of biliary structures and liver at laparoscopy followed by cholangiograms and anatomical dissection. Five patients had ultrasonography of the biliary tract at laparoscopic cholecystectomy. RESULTS: All animals had adequate visualization of important hepatobiliary structure, and an optimal method of accessing these structures at laparoscopy was established. Patients had ultrasonography which used methods developed in the animal trial. All had adequate visualization of the entire common bile duct confirmed by cholangiography. Limitations in demonstrating the relationship of the cystic duct to the common duct were technical and can be corrected. CONCLUSION: Laparoscopic ultrasonography has significant potential for delineation of biliary anatomy and determination of presence or absence of duct calculi. Clinical implementation could minimize the risk of iatrogenic duct injury and the need for operative cholangiography.

Animals↗
PubMedDate not supplied

Monitor peptide (rat pancreatic secretory trypsin inhibitor) directly stimulates the proliferation of the nontransformed intestinal epithelial cell line, IEC-6.

BACKGROUND/AIMS: The growth-stimulating activity of monitor peptide [rat pancreatic secretory trypsin inhibitor (PSTI)-61] was evaluated in the rat intestinal epithelial cell line IEC-6. In response to food intake, monitor peptide induces cholecystokinin release and the subsequent secretion of pancreatic enzyme into the rat small intestine. METHODS: Monitor peptide was purified from rat bile-pancreatic juice. The proliferation of IEC-6 cells was determined by both counting cell number and [3H]-thymidine incorporation. Ornithine decarboxylase (ODC) activity was assessed by radiometric technique using L-[1-14C]-ornithine. RESULTS: Monitor peptide (10(2)-10(5) ng/ml) induced a significant increase in the incorporation of [3H]-thymidine into IEC-6 cells, and this effect was observed in a dose-dependent manner. A significant increase in the cell number was also observed. An antibody specific for the synthetic NH2-terminal fragment of monitor peptide completely abrogated the growth-stimulating effects of this peptide. In addition, monitor peptide effectively increased the ODC activity of IEC-6 cells. CONCLUSIONS: These results indicate that monitor peptide stimulates the growth of intestinal epithelial cells, and suggest that polyamine metabolism may be involved in this mechanism of growth induction.

Animals↗