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Biomedical subjects

A Ooshima

Publications and source records attributed to A Ooshima.

At least 109 records · Page 6Linked to original sources
PubMed1990

One-step sandwich enzyme immunoassay for human laminin using monoclonal antibodies.

A one-step sandwich enzyme immunoassay (one step sandwich EIA) for human serum immunoreactive laminin was set up with a pair of monoclonal antibodies prepared against human placental laminin P1 fragment. The assay was characterized by carrying out two immunoreactions simultaneously, laminin P1 fragment reacting with both a monoclonal antibody as a solid phase and a horseradish peroxidase-labeled monoclonal antibody (Fab') against human laminin P1 fragment as conjugate. Sensitivity of the immunoassay was 0.01 ng/well (0.5 microgram/l), and linearity was obtained between 0.01-20 ng/well (0.5-1,000 micrograms/l). The levels of laminin in sera from normal individuals and patients with liver cirrhosis, hepatocellular carcinoma and primary biliary cirrhosis were 103 +/- 15 micrograms/l, 228 +/- 70 micrograms/l, 341 +/- 163 micrograms/l and 232 +/- 93 micrograms/l, respectively. Protein immunoblotting showed that the serum immunoreactive laminin measured by the assay was a fragment with rel mol mass of 200 kDa.

Antibodies, Monoclonal↗
PubMed1990

Synthesis of lysyl oxidase in experimental hepatic fibrosis.

The synthesis of lysyl oxidase, which initiates the cross-linking of collagen and elastin, was investigated in carbon tetrachloride (CCl4) induced fibrotic liver of rat. Lysyl oxidase activity of the fibrotic liver was 4 times greater than that of normal liver. mRNAs from the livers of normal and CCl4-treated rats were prepared for in vitro protein synthesis and the products were analyzed by immunoprecipitation with a monoclonal antibody against lysyl oxidase. The mRNAs from the fibrotic liver gave more than 3 times higher level of messenger copies for lysyl oxidase than did mRNAs from normal liver. The molecular weight of the nascent lysyl oxidase was 48,000.

Amino Acid Oxidoreductases↗
PubMed1990

Distribution of type I collagen in human kidney diseases in comparison with type III collagen.

The distribution of type I collagen in normal and diseased renal tissues was studied using immunofluorescence and immunoelectron microscopy, and was compared with that of type III collagen. In normal human kidneys, a monoclonal antibody against type I or type III collagen reacted with the renal interstitium, but not with the intra-glomerular structures. In various types of glomerulonephritis, immunofluorescent staining for type I collagen was positive in the fibrocellular and fibrous crescents, sclerosed glomeruli, and infrequently within the glomerular mesangium. In the crescents and sclerosed glomeruli, type I collagen was co-localized with type III collagen. The staining intensity of type I collagen in those areas was generally stronger than that in the interstitium. Mesangial staining for type I collagen was present within the glomeruli, particularly with a marked mesangial matrix increase, but was less in amount and frequency compared with type III collagen staining. These findings indicate that the fibrosclerotic process in damaged glomeruli is accompanied by the appearance of interstitial collagens, and that participation of type I collagen is prominent in crescent organization and global glomerular sclerosis, but is less frequent in mesangial expansion, compared with type III collagen.

Adolescent↗
PubMed1990

Immunoelectron microscopic localization of human type III collagen in human gastrointestinal carcinomas.

Ultrastructural immunolocalization of human type III collagen/procollagen was investigated in 28 cases of human gastrointestinal carcinoma by the pre-embedding method using a monoclonal antibody. In addition to immunoreactivity on collagen fibers, amorphous or finely fibrillar immunoreactive substances were observed in the extracellular matrix adjacent to the plasma membrane and in the cisternae of rough endoplasmic reticulum (rER). These substances were considered to represent type III procollagen and/or collagen before fiber formation. Cytoplasmic recesses containing immunoreactive fibers were also observed. These findings related to collagen production were detected in stromal fibroblasts in 16 of 28 cases and in carcinoma cells in six of 28 cases. The positivity rate of these findings was lower in diffuse-type gastric carcinoma. The present study thus clarified further details of collagen production and secretion, and revealed that carcinoma cells may also be involved in the production of stromal collagen in some cases as well as stromal fibroblasts.

Antibodies, Monoclonal↗
PubMed1990

Primary cutaneous adenoid cystic carcinoma: ultrastructural study and immunolocalization of types I, III, IV, V collagens and laminin.

A primary adenoid cystic carcinoma of the skin is reported. Light microscopy revealed pseudocysts. PAS-positive basement membrane and true glandular lumen, which in aggregates are specific for adenoid cystic carcinoma. Perineural invasion was also observed. Ultrastructural examinations revealed three types of cystic spaces; pseudocysts, true glandular lumens and intercellular spaces. Enzyme histochemical examinations showed positive reactions for eccrine enzymes, including phosphorylase and succinic dehydrogenase and negative for apocrine enzymes. Immunolocalization of collagens and laminin revealed that basement membranes of the pseudocysts involve Type V collagen as well as Type IV collagen and laminin.

Aged↗
PubMed1990

Immunohistochemical localization of pituitary gonadotrophins and gonadal steroids confirms the 'two-cell, two-gonadotrophin' hypothesis of steroidogenesis in the human ovary.

Ovaries from 37 women with normal menstrual cycles were analysed for localization of pituitary gonadotrophins and gonadal steroids using an immunohistochemical method. In the follicular phase, FSH and oestradiol-17 beta localized in the granulosa layer, and LH, progesterone and testosterone localized in the internal thecal layer. In the luteal phase, gonadotrophins and steroids localized in luteal cells. Particularly in the early luteal phase, FSH and oestradiol-17 beta localized in large luteal cells, and LH, progesterone and testosterone localized in small luteal cells. The results of the present immunohistochemical analysis confirm the two-cell, two-gonadotrophin hypothesis of steroidogenesis in the human ovary.

Adult↗
PubMed1990

Serum type IV collagen concentrations in diabetic patients with microangiopathy as determined by enzyme immunoassay with monoclonal antibodies.

In long-term diabetes mellitus, thickening of basement membrane in capillaries and small vessels is a characteristic lesion and plays an important role in the progression of diabetic microangiopathy. We have developed a sandwich enzyme immunoassay for human serum type IV collagen peptide with monoclonal antibodies. Previous studies suggested that collagen levels reflect the activity of fibrogenesis in basement membrane. Serum type IV collagen levels were measured in 137 non-insulin-dependent diabetic patients (aged 50-75 yr) with or without clinical signs of retinopathy, nephropathy, and/or neuropathy and 110 healthy subjects (aged 50-75 yr) without serological abnormality. Serum concentrations of type IV collagen were significantly higher (P less than 0.01) in diabetic patients (mean +/- SE 124.1 +/- 4.1 ng/ml) than in healthy subjects (73.9 +/- 2.2 ng/ml) and were increased with the prevalence or incidence of diabetic complications. In the patients with diabetic microangiopathy, serum type IV collagen levels became higher as clinical signs worsened. Especially in the patients with diabetic nephropathy, serum type IV collagen levels became higher with elevation of blood urea nitrogen, serum creatinine, and serum beta 2-microglobulin but not urinary excretion of beta 2-microglobulin and N-acetyl-beta-glucosaminidase. These observations indicated that elevation of serum type IV collagen in diabetic nephropathy was related to glomerular filtration dysfunction rather than renal tubular dysfunction. However, the antigen, which can be detected by our assay system, did not exist in urine specimens of healthy subjects, and an intimate relationship was not observed between serum type IV collagen level and serum creatinine level.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1990

Possible formation of cutaneous amyloid from degenerative collagen fibers. Ultrastructural collagen changes and the immunoreactivity of cutaneous amyloidosis employing anti-type I, III, IV, V collagen antibodies.

The origins of primary cutaneous amyloid have been investigated and several possibilities are proposed. We investigated here several cases by electron microscopy and found in all cases characteristic filamentous changes which were identical with amyloid structures in the collagen bundles. These observations suggested a possible pathway of cutaneous amyloid formation from degenerative collagen fibers. Further studies were performed using immunoelectron microscopical and immunohistochemical methods employing a panel of anti-collagen antibodies to examine the reactivity of amyloid to them. Our results were much more suggestive of the collagen origin of this kind of amyloid.

Amyloid↗
PubMed1990

Immunohistochemical localization of lysyl oxidase with monoclonal antibodies.

Lysyl oxidase initiates the cross-linking of collagen and elastin by catalyzing the formation of the lysine-derived aldehyde. We cloned three hybridoma cell lines which secrete monoclonal antibodies to human lysyl oxidase. The localization of lysyl oxidase was investigated in various tissues and in cultured cells using an immunofluorescent antibody method. Antibodies showed a strong immunostaining in the aorta and dermal connective tissue suggesting a close relation to elastin and collagen. Fibroblasts, chondrocytes, and smooth muscle cells also yielded a marked positive immunoreaction as did a variety of nonfibroblastic cells such as endothelial cells, basal cells, biliary epithelial cells, and glomerular epithelial cells. In cultured cells, including human fibroblasts, an intense immunoreaction manifested as fine, filamentous structures in the cytoplasm. It is suggested that lysyl oxidase is associated with cytoskeletal protein.

Amino Acid Oxidoreductases↗
PubMed1990

Primary glomerular fibrosis: a new nephropathy caused by diffuse intra-glomerular increase in atypical type III collagen fibers.

A 38-year-old Japanese man suffering from nephrotic syndrome and slowly progressive impairment of renal function underwent renal biopsy. Light microscopy revealed diffuse lobular accentuation of glomerular tufts, occupied by homogeneous material which was eosinophilic, lightly stained with periodic acid silver methenamine and deeply with aniline blue. Further examinations using an electron microscope after treatment with phosphotungstic acid revealed that this material exhibited a diffuse increase in fibers having a periodicity of 65 +/- 4 nm (mean +/- SD) like type III collagen fibers and was reactive to the anti-type III collagen monoclonal antibody. In addition, the serum level of procollagen III peptides was markedly elevated to 221 ng/dl (normal range 2-13 ng/dl). However, the fibers were peculiarly curved and frayed as reported in the nail-patella syndrome, but were not observed in the lamina densa of the glomerular basement membrane (GBM) as seen in this syndrome. No family history or physical findings of bony and nail dystrophy, no laboratory data or pathological findings compatible with already described renal diseases including mesangiocapillary glomerulonephritis, light chain disease, immunotactoid nephropathy, amyloidosis, and non-amyloidotic fibrillary glomerulonephritis were observed. These data strongly suggest that the nephropathy is caused by a primary increase in atypical type III collagen fibers in glomeruli (primary glomerular fibrosis), which to our knowledge has not yet been described in the English literature.

Adult↗
PubMed1989

One step sandwich enzyme immunoassay for human type IV collagen using monoclonal antibodies.

Monoclonal antibodies were used in one step sandwich enzyme immunoassay (one step sandwich EIA) for human serum immunoreactive type IV collagen. The one step sandwich EIA using either polystyrene ball or microplate was characterized by carrying out two immunoreactions simultaneously, type IV collagen reacting with both a monoclonal antibody as a solid phase and a horseradish peroxidase-labeled monoclonal antibody (Fab') against human type IV collagen as a conjugate. Sensitivity of one step sandwich EIA system by using either polystyrene ball or microplate was 0.22 ng per tube or 0.04 ng per well for type IV collagen, and linearity was obtained between 0.22-40 ng/tube or 0.04-20 ng per well, respectively. Both methods gave reproducible quantitative analysis of immunoreactive type IV collagen levels in the sera of patients with hepatocellular carcinoma and patients with liver cirrhosis, which were apparently higher than the levels in the sera of healthy subjects. Protein immunoblotting shows that the immunoreactive type IV collagen trapped in our present one step sandwich EIA system was not the 7-S and NC1 domains of type IV collagen.

Antibodies, Monoclonal↗
PubMed1989

Alterations in the extracellular matrix components in human glomerular diseases.

We investigated the distribution of extracellular matrix components such as fibronectin, laminin, type III, IV, V, and VI collagens and heparan sulfate proteoglycan (HSPG) in normal and diseased glomeruli using the indirect immunofluorescence method. This study included 96 renal biopsies: 7 controls, 3 minimal change nephrotic syndrome (MCNS), 47 mesangial proliferative glomerulonephritis (PGN), 25 membranous nephropathy (MN) and 14 membranoproliferative glomerulonephritis (MPGN) including 3 lupus nephritis. Fibronectin was detected predominantly in the mesangium and less prominently in the glomerular basement membrane (GBM) of normal glomeruli. Laminin and type IV collagen were present in the mesangium and GBM, type III collagen in the interstitium, and type V collagen in the mesangium, interstitium and a part of GBM. Type VI collagen was observed in the mesangium, interstitium and slightly in GBM. Anti-HSPG antibody reacted with the mesangium and GBM. MCNS showed a distribution of these antigens similar to that in normal controls. The finding that staining for HSPG was not decreased in the GBM and mesangium indicated that there was no change in the core protein of HSPG. Fibronectin, laminin, type IV collagen and HSPG were increased in the thickened GBM of MN and in the expanded mesangium of PGN. In MPGN, these matrix components were increased in the mesangium and GBM with remarkable increase of type V and VI collagens. While type III collagen was not found in normal glomeruli, it became detectable in the mesangium and a part of GBM in MPGN. No significant decrease in the intensity of fluorescence for HSPG was observed in the glomeruli from nephrotic patients. These findings suggest that proteinuria might be caused by the structural alteration in the glycosaminoglycan portion of HSPG, changes in any anionic material other than HSPG, or both, and also indicate that the glomerular mesangial sclerosis is closely related to the increase of type V and VI collagens.

Basement Membrane↗
PubMed1989

Localization of gonadotropin binding sites in human ovarian neoplasms.

The binding of human luteinizing hormone and human follicle-stimulating hormone to ovarian tumor biopsy specimens from 29 patients was analyzed. The binding sites for human luteinizing hormone were demonstrated in one tumor of epithelial origin (mucinous cystadenoma) and in one of sex cord-stromal origin (theca cell tumor). The binding sites for human follicle-stimulating hormone were found in three tumors of epithelial origin (serous cystadenoma and mucinous cystadenoma) and in two of sex cord-stromal origin (theca cell tumor and theca-granulosa cell tumor). The surface-binding autoradiographic study revealed that the binding sites for gonadotropins were localized in the stromal tissue. The results suggest that gonadotropic hormones may play a role in the growth and differentiation of a certain type of human ovarian neoplasms.

Adolescent↗
PubMed1989

Glomerular localization of type III collagen in human kidney disease.

Kidney specimens taken from normal humans and patients with various renal diseases were examined by immunofluorescent and immunoelectron microscopy using a monoclonal antibody to the alpha 1 chain of type III collagen. Indirect immunofluorescent staining revealed that intraglomerular localization of type III collagen antigen in 41 of 66 patients, while it was absent from the glomeruli of normal human kidneys. Type III collagen was found within the mesangium of 22 patients with various types of renal diseases, and was distributed in a focal and segmental manner in most of the cases. Mesangial localization of the collagen correlated with the increase in the mesangial matrix. Type III collagen was also present in the vascular pole, crescents (particularly in the organizing phase) and sclerosed glomeruli. Immunoelectron microscopy using pre-embedding and post-embedding techniques confirmed the above observations. These findings indicate that type III collagen participates in mesangial expansion, crescent organization, and glomerulosclerosis.

Antibodies↗
PubMed1989

The subbasement membrane distribution of type IV collagen in normal human skin.

Samples of normal human skin were obtained from 48 sites in 26 subjects ranging in age from 2 to 85 years. The samples were examined by indirect immunofluorescence and immunoelectron microscopy using anti-human type IV collagen antibodies produced by immunizing rabbits with type IV collagen extracted from human placenta. Fluorescence was observed as granular or fine fibrous patterns, not only in the basement membrane at the dermo-epidermal junction, around the vessels, and the accessory organs of the skin, but also in the dermal regions in the vicinity of the basement membranes. This suggests the presence of type IV collagen in the dermis deep to the basement membrane. Ultrastructurally, the extrabasal lamina distribution of type IV collagen was noted as a partial distribution around the fibroblasts that existed close to the basal lamina. These findings are considered to be important in examining the function of this collagen in the dermis and the dynamics and metabolism of the basement membrane under normal and abnormal conditions.

Adolescent↗
PubMed1989

Increased amount of serum type IV collagen peptide in human liver fibrosis as determined by enzyme-immunoassay with monoclonal antibodies.

Chronic liver injury by various toxic agents causes an increase in collagen biosynthetic activity, resulting in deposition of excessive amounts of collagen and rearrangement of the lobular architecture leading to hepatocellular dysfunction and portal hypertension. The authors have developed sandwich enzyme-immunoassay for human serum type IV collagen peptide using monoclonal antibody as a marker of fibrogenesis and examined the relationship between the amount of this collagen peptide and hepatic disorders including chronic active hepatitis and liver cirrhosis. Sera from patients with chronic active hepatitis or liver cirrhosis were used after confirmation of the histopathological diagnosis. Control sera were obtained from healthy subjects without any serological abnormality in liver function tests. Serum type IV collagen peptide levels were significantly higher in patients with chronic hepatic disorders associated with fibrosis than in healthy subjects. This non-invasive enzyme-immunoassay gave reproducible quantitation, and serum type IV collagen peptide was concluded to be a useful, and reproducible marker for the early detection of fibrogenesis in the liver.

Antibodies, Monoclonal↗
PubMed1989

Increased serum type IV collagen peptide in carbon tetrachloride-treated rats.

We developed a competitive enzyme-immunoassay for serum type IV collagen peptide as a marker of fibrogenesis, and examined the relationship between serum type IV collagen peptide and hepatic disorder in CCl4-treated rats. The rats were treated for 8 weeks and signs of liver damage began to appear from about week 2. With the progression of these signs to liver fibrosis, type IV collagen increased in the fibrous septa and especially in the perisinusoidal walls, where the increase was manifested as development of a real basement membrane beneath the sinusoidal endothelial cells. In CCl4-treated rats, serum type IV collagen peptide significantly increased with the progression of liver fibrosis. When CCl4 administration was stopped, the collagen peptide rapidly decreased without any rebound rise. An intimate relationship was found between the production of serum type IV collagen peptide and liver prolyl hydroxylase activity and the amount of collagen deposited in the liver. These results suggest that serum type IV collagen peptide will be a useful biochemical marker for the early detection of fibrogenesis in the liver.

Animals↗