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Biomedical subjects

K Nabeshima

Publications and source records attributed to K Nabeshima.

At least 91 records · Page 5Linked to original sources

Jejunal perforation associated with cytomegalovirus infection in a patient with adult T-cell leukemia-lymphoma.

A patient with adult T-cell leukemia-lymphoma suffered a jejunal perforation, which we believe was directly attributable to cytomegalovirus (CMV) infection. In the areas of ulceration and perforation in the small bowel, blood vessels penetrating the muscularis propria showed extensive lining of cytomegalic endothelial cells with CMV inclusions, accompanied by occasional disruption of the walls, partial occlusion of the lumina, fibrin thrombi, and hemorrhage. The CMV-induced vascular damage seemed to be closely related to the occurrence of ulcers and perforation. The recognition of CMV as a cause of lethal gastrointestinal lesions in immunocompromised hosts has become more important with the advent of anti-CMV therapy.

Adult↗

Antilymphocyte antibodies against CD4+CD45R+ subsets in patients with IgA nephropathy.

The authors analysed CD4+ subset populations and particularly subset killing in order to evaluate the presence of antilymphocyte antibody against CD4+ subsets in patients with IgA nephropathy (IgA N). This study was performed in 45 patients with IgA N, 30 patients with other forms of glomerulonephritis and 30 healthy controls. CD4+, CD4+CD45R- and CD4+CD45R+ cells in the peripheral blood were counted by a flow cytometric analysis, and those cell killings were analysed by microcytotoxic assays. The percentage of circulating CD4+CD45R+ cells was significantly decreased in IgA N, and the percentage of CD4+CD45R+ cell killing was significantly elevated in IgA N compared with other groups. There was a significant negative correlation between the percentage of CD4+CD45R+ cells present in IgA N patients' peripheral blood lymphocytes and the killing CD4+CD45R+ cells by the same patients' serum. Both a depletion of CD4+CD45R+ cells in peripheral blood lymphocytes and an elevation of CD4+CD45R+ cell killing correlated with the grade of mesangial proliferation in patients with IgA N. However, there were no correlations in other clinicopathological indices. These results suggest that low levels of antilymphocyte antibodies against CD4+CD45R+ cells were present in patients with IgA N, who showed a depletion of CD4+CD45R+ cells in the peripheral blood. These antibodies were strongly associated with the elimination of CD4+CD45R+ cells and the proliferation of glomerular mesangial cells in patients with IgA N.

Adolescent↗

Establishment of a new human cancer cell line secreting protease nexin-II/amyloid beta protein precursor derived from squamous-cell carcinoma of lung.

A new cell line (LC-1/sq) of human lung squamous-cell carcinoma was established from a surgically resected specimen of primary lung cancer. Upon continuous propagation in serum-free culture medium, it secreted trypsin inhibitors into the conditioned medium. The major fraction of the trypsin inhibitor (T1-1) was purified to apparent homogeneity by anion-exchange and gel-filtration high-performance liquid chromatography (HPLC) and sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by transblotting to Immobilon. T1-1 effectively inhibited trypsin. Chymotrypsin, plasmin and kallikrein were inhibited to a lesser extent, but urokinase-type plasminogen activator, elastase, thrombin and papain were not inhibited. The activity of T1-1 was acid-stable and heat-resistant, and its molecular weight was 115 kDa by SDS-PAGE. It exhibited single NH2-terminal sequence, and its first 20 NH2-terminal amino-acid residues were identical with those of protease nexin-II (PN-II)/amyloid beta-protein precursor (APP). These characteristics of T1-1 suggest that the major trypsin inhibitor secreted by LC-1/sq is indistinguishable from PN-II/APP. LC-1/sq is the first lung squamous carcinoma cell line that secretes functionally active trypsin inhibitor, PN-II/APP, in vitro and is useful for studying its biological significance in malignant tumor.

Aged↗

Partial sequencing and characterization of the tumor cell-derived collagenase stimulatory factor.

A tumor cell-derived, collagenase stimulatory factor (TCSF), previously isolated and purified from LX-1 human lung carcinoma cells and judged by immunoblotting and SDS-PAGE to contain a single protein of approximately 58 kDa, has been further analyzed for its biological activity and composition. Three significant new findings have been made. First, the biological activity of TCSF preparations was shown definitively to reside in the 58-kDa protein. This was achieved in two ways: (a) a polyclonal antibody was raised against the 58-kDa protein, after excision from an SDS-PAGE gel, and shown to inhibit the stimulation of fibroblast collagenase production by TCSF preparations; (b) the 58-kDa protein was eluted from a transblot of purified TCSF and shown to stimulate fibroblast collagenase production. Second, partial sequencing of the 58-kDa protein revealed no significant homologies with other known collagenase stimulatory factors. Third, purified TCSF was found, on transblotting to Immobilon, to contain a doublet of 58 kDa (TCSF1) and 54 kDa (TCSF2) proteins; the former was present in higher concentration than the latter. N-terminal amino acid sequencing of the two intact proteins and of four corresponding pairs of tryptic peptides derived from the two proteins showed identity in each case, indicating that TCSF1 and TCSF2 are very similar in composition. However, TCSF1 but not TCSF2 stimulated fibroblast collagenase production, confirming that the 58-kDa protein is the major active component of TCSF preparations.

Amino Acid Sequence↗

Glycocalyceal bodies in a human rectal carcinoma cell line and their interstitial collagenolytic activities.

In co-cultivation on a membrane of connective tissue matrix (CTM) obtained from human dura mater, human adenocarcinoma cells (RCM-1) degraded CTM. Morphologically, the destruction of CTM was associated with the shedding of membrane vesicles from the cells. Transmission electron microscopy, using ruthenium red (RR), showed that the shed vesicles were composed of various-sized membrane bound vesicles (MV). A large majority were small glycocalyceal bodies (G-bodies) measuring 20-120 nm in diameter, composed of an amorphous matrix of moderate electron-density surrounded by an RR-positive, trilaminar membrane. G-bodies were separated from medium-sized and large MVs by ultracentrifugation. Ultrastructural observation of the isolated collagen fibrils from CTM co-cultured with RCM-1 cells, showed G-bodies attached to degraded collagen fibrils with characteristic transverse notches along their axes. The lesions occurred as microerosions in the apolar region between the e and d bands of collagen fibrils. Collagenolytic activity in serum-free RCM-1 conditioned medium was localized in the G-body and MV fractions (80% and 20% of the total activity, respectively, when tested against 3H-labeled type I collagen). No activity was detected in the supernatant. The activity in G-bodies was also confirmed by ultrastructural analysis using reconstituted native type I collagen fibrils. The results suggest that RCM-1 cells release interstitial collagenase as a component of G-bodies which facilitates local breakdown of connective tissue during the process of invasion.

Adenocarcinoma↗

[A case of adult T-cell leukemia with a defective HTLV-I proviral DNA, in which the single T-cell clone appeared to have progressed from chronic phase to crisis].

A 39-year-old woman was first admitted to our hospital with increased white cell count on May, 1983. Physical examination showed only mild splenomegaly. Hematological examination revealed leukocytosis (14,600/microliters) with ATL cells (59%). Serum anti-HTLV-I antibody was positive. Examination of HTLV-I provirus in the abnormal T cells revealed the defective type. She was diagnosed as chronic type of ATL based on the clinical features. Cytogenetic study of the ATL cells revealed 47, xx, +4. For 12 months, she was followed without any therapy. WBC reduced to almost normal range and ATL cells decreased to 3 to 6% for 8 months. On May, 1985, she was readmitted to our hospital because of leukocytosis (32,200/microliters), and increased ATL cells (57%). She was diagnosed as crisis of ATL. Investigation of the proviral DNA and chromosome showed the same results as those of the chronic phase, indicating that ATL cells in both the chronic phase and the crisis phase originated from the same clone. She died after 3 months from massive diarrhea. Postmortem examination showed the extensive infiltration of leukemic cell in the small intestine.

Adult↗

[The effects of r-HuEPO on platelet function and coagulation factors in hemodialysis patients].

To evaluate the effects of correction of anemia with recombinant human erythropoietin (r-HuEPO) on the hemostatic defects in uremia, hemostatic parameters were examined in 18 patients with renal anemia receiving hemodialysis (HD). During the study, hematocrit (Ht) increased from 22.9 +/- 3.1% (mean +/- SD) at pre-treatment (stage-I) to 31.0 +/- 3.0% 12 weeks after 3000 IU intravenous r-HuEPO administration at the end of every HD (stage-II), and decreased to 26.2 +/- 4.2% 6 weeks after r-HuEPO discontinuation (stage III). Platelet count did not change among these three stages, however, mean platelet volume significantly increased at stage II compared to stage I. Ivy bleeding time (Ivy-BT) significantly shortened at stage II (I; 14.3 +/- 6.0, II; 10.1 +/- 6.5 min, p less than 0.01), and prolonged again at stage III (p less than 0.05 vs stage II). Among the patients, 6 out of 18 patients did not show any reduction in Ivy-BT (unchanged group). Though there were no significant changes in platelet aggregation rates, plasma TxB2, 6-keto-PGF1 alpha, F. VIII: C, and F. VIII: Ag levels throughout the study, platelet adhesion rate was significantly improved at stage II (I; 11.8 +/- 6.8, II; 19.6 +/- 12.8%, p less than 0.05), and similar augmentation in vWf: Ag was observed. Improvement in these two parameters were more remarkable in shortened Ivy-BT group (n = 12) than in unchanged group.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Monoclonal antibody preparation and purification of a tumor cell collagenase-stimulatory factor.

Tumor cells from several sources produce a factor(s) which stimulates fibroblast collagenase production. Monoclonal antibodies have been raised against the tumor cell collagenase-stimulatory factor from LX-1 human lung carcinoma cells and have been used for purification of the factor from LX-1 cell membranes. These purified preparations stimulated fibroblast collagenase production, and 80% of these preparations contained a single Mr approximately 58,000 protein detectable by immunoblotting; the other 20% contained an additional minor component with a molecular weight of 35,000. A single protein with a molecular weight of approximately 58,000 was also detected in radiolabeled preparations of the purified factor by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Conditioned media from LX-1 cells contain several species with molecular weights lower than 58,000 which are immunologically cross-reactive with the membrane-derived factor. Immunofluorescence analysis indicates that the tumor cell collagenase-stimulatory factor is distributed on the outer surface of LX-1 cells and is absent from the cell surface of fibroblasts. These and previous results indicate that the factor is present on the tumor cell surface, is released into conditioned media possibly after proteolytic cleavage, and appears to have an important role in inducing collagenolysis of host stroma during tumor invasion.

Animals↗

[Clinical significance of the intranuclear cleft of the lumbar intervertebral disk on MRI].

MR studies of the lumbar spine in 111 patients were analyzed at 469 disks to assess the prevalence of intranuclear cleft (INC) in the lumbar intervertebral disk. MR studies were performed on either 0.1-tesla (T) magnet (69 patients) or 0.22-T magnet (42 patients). The pulse sequences reviewed were saturation recovery (SR; TR = 0.5 sec), short TR, TE spin echo (S-SE; TR = 0.5 sec, TE = 34 msec) and long TR, TE spin echo (L-SE; TR = 1.5 sec, TE = 68,80 msec). All study were done in a sagittal plane with 10 mm slice thickness. The conclusions were as follows: 1) On a 80 msec TE, 1.5 sec TR image, INCs were detected in more than 80% of disks in patients over 30 years old but in only 13.3% of disks in patients under 20 years old. 2) In both imaging system, L-SE showed INCs more frequently than SR and S-SE. 3) INCs were less frequently demonstrated in the disk with decreased signal intensity on 0.1-T magnet as compared with 0.22-T magnet. 4) On SR and S-SE, there is an increase in the prevalence of INC in the disk with decreased signal intensity. We suggest that the INC will be a good landmark of the pathological process of the lumbar disk, such as degeneration.

Adolescent↗

New human colorectal carcinoma cell lines that secrete proteinase inhibitors in vitro.

Two new human cell lines, RCM-1 and CoCM-1, have been established from primary colorectal adenocarcinomas. Both cell lines were unique in that the cultures secreted trypsin inhibitors in vitro. The activities of these inhibitors were accumulated in serum-free media of both cell lines over a period of several days. Two inhibitors (PI-1 and PI-2) were isolated from serum-free conditioned medium in which RCM-1 was grown by anion-exchange and gel filtration high-performance liquid chromatography. PI-1 inhibited trypsin and chymotrypsin strongly, and pancreatic elastase weakly. Its molecular weight was about 57 kilodaltons (Kd) as determined by gel filtration chromatography. It cross-reacted with the antiserum elicited against human alpha 1-antitrypsin in double immunodiffusion. PI-1 corresponding to alpha 1-antitrypsin was also demonstrated immunohistochemically in both cell lines. PI-2 inhibited trypsin strongly, and chymotrypsin, kallikrein and plasmin weakly. It had higher molecular weight (200-300 Kd) than that of PI-1, and did not cross-react with antisera against human alpha 1-antitrypsin, alpha 2-macroglobulin, alpha 1-antichymotrypsin, alpha 2-plasmin inhibitor, inter-alpha-trypsin inhibitor and urinary trypsin inhibitor. RCM-1 and CoCM-1 are the first colorectal adenocarcinoma cell lines that secrete functionally active trypsin inhibitors, including alpha 1-antitrypsin in vitro, and are useful for the study of tumor-cell derived proteinase inhibitors.

Adenocarcinoma↗

Tumor markers in pleural effusion diagnosis.

In order to discriminate between malignant and benign effusions, the values of carcinoembryonic antigen (CEA), ferritin, beta2-microglobulin (BMG), acid-soluble glycoprotein (ASP), tissue polypeptide antigen (TPA), adenosine deaminase (ADA), and immunosuppressive acidic protein (IAP) were measured in the pleural fluid of 54 patients with lung cancer, 20 with malignancies other than lung cancer, 18 with tuberculous pleurisy, and 22 with benign diseases other than tuberculosis. CEA levels in malignant effusions were significantly higher than those in benign effusions. At a cutoff level of 5 ng/ml, 68% of the patients with lung cancer and 44% of the patients with other malignancies showed elevated pleural fluid CEA levels. In 13 lung cancer cases with negative pleural fluid cytology, nine cases had elevated pleural fluid CEA levels. The mean pleural fluid BMG level of patients with benign diseases was significantly higher than that of patients with malignant diseases, but there was a marked overlap between those with malignant and benign diseases. No significant differences were found in the pleural fluid ferritin, ASP, TPA, and IAP levels between malignant and benign conditions. ASP and IAP pleural fluid levels showed significant correlations with the pleural fluid C-reactive protein (CRP) concentrations suggesting that they also reflect inflammatory activity. The mean ADA activity in tuberculous effusion was significantly higher than that resulting from other causes of pleural effusion.

Adult↗

High-molecular-weight neutral proteinase of rat ascites hepatoma cells and their collagenolytic activity.

A high-molecular-weight proteinase in rat ascites hepatoma AH109A cells was purified to apparent homogeneity by conventional chromatographic techniques. The purified proteinase degraded over the broad pH range, 7.0-9.0, not only denatured hemoglobin but also type I and IV collagen in the absence of calcium. Its activity was maximum at pH 8.0, heat-labile and affected by thiol reagents. Serine proteinase inhibitors such as diisopropyl fluorophosphate, soybean trypsin inhibitor, leupeptin, and antipain also partially inhibited its activity. The enzyme was found mainly in the cytosol fraction, and had a molecular weight of 450 kilodaltons (Kd) as determined by gel filtration chromatography and showed a single band on polyacrylamide gel electrophoresis under nondenaturing condition, although it was dissociated into lower-molecular-weight subunits, 25.5-32 Kd in the presence of sodium dodecyl sulfate and 2-mercaptoethanol. These properties suggested that it was a kind of a cytosolic high-molecular-weight proteinase. The collagenolytic activity of this proteinase may play an important role in cancer cell invasion and metastasis.

Animals↗

A new long-term in vitro invasion assay using fibrous connective tissue matrices maintaining architectural characteristics of connective tissue.

A long-term invasion assay using fibrous connective tissue matrices was developed. The matrices were prepared by treating murine skin or human dura mater with 25 mM ammonium hydroxide containing proteinase inhibitors at 4 degrees C for 7 days. They could be maintained almost indefinitely without the degeneration and necrosis. Electron micrographs of them revealed the preservation of native collagen fibers, and sequential enzyme digestion showed the presence of glycoprotein in the matrices. Local dissolution of extracellular matrix by cultured human rectal adenocarcinoma cell line, RCM-1, was observed morphologically and confirmed by a quantitative assay using radiolabeled matrices. The destruction of extracellular matrix occurred associated with membrane vesicle-shedding from the cells. Both the advantages and disadvantages of this assay were discussed.

Animals↗

Erythrocyte 2,3-diphosphoglycerate in liver diseases.

The erythrocyte 2,3-diphosphoglycerate (2,3-DPG) was studied in patients with liver diseases, chronic obstructive pulmonary disease, and in normal subjects. The level of 2,3-DPG in liver diseases occurred in the following increasing order: chronic persistent hepatitis, chronic active hepatitis, liver cirrhosis, and cirrhosis with hepatocellular carcinoma. A significant negative correlation between the 2,3-DPG concentration and serum albumin concentration was found in the liver diseases. The 2,3-DPG level was correlated to the serum concentration of total bile acids and to the arterial blood pH. A negative correlation was found between the arterial blood pH and the serum albumin concentration. The level of 2,3-DPG in hepatocellular carcinoma and/or liver cirrhosis was higher than that in more hypoxic chronic obstructive pulmonary disease. And an increased level of 2,3-DPG was also shown in nonhypoxic patients with liver diseases. These results suggest that the level of erythrocyte 2,3-DPG increases according to the severity of the liver disease, and compared to the level in hypoxic chronic obstructive pulmonary disease, the level of erythrocyte 2,3-DPG is higher in both hepatocellular carcinoma and liver cirrhosis.

2,3-Diphosphoglycerate↗