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Biomedical subjects

M Mizuno

Publications and source records attributed to M Mizuno.

At least 19 recordsLinked to original sources

Effects of a bone lysine-rich 18 kDa protein on osteoblast-like MC3T3-E1 cells.

A lysine-rich 18 kDa protein was isolated from bovine bone and examined for its effects on osteoblast-like MC3T3-E1 cells. This protein is homologous to a heparin-binding protein in brain and uterus. This protein enhanced cell attachment independent of the Arg-Gly-Asp cell-binding sequence and stimulated proliferation during the growth phase. Addition of this protein to cell cultures on days 11, 12, and 13 after confluency resulted in a 1.6-2.0-fold increase in the alkaline phosphatase activity and little increase in the DNA content. These findings suggest that the 18 kDa protein may be functional in promoting the proliferation and differentiation of osteoblasts.

3T3 Cells

Properties of plasma membrane-induced amylase release from rat parotid secretory granules: effects of Ca2+ and Mg-ATP.

A secretory granular fraction (SG) and a plasma membrane rich fraction (PM) have been isolated from rat parotid gland by differential and Percoll gradient centrifugation. With these two fractions, a cell-free interaction system has been reconstituted to clarify the exocytotic interaction between the secretory granules and plasma membranes, and the conditions of amylase release from SG have been characterized in vitro. The addition of PM into this assay system induced a rapid and transient release of amylase from SG. Some other membranes such as erythrocyte ghosts also mimicked the effect of PM. This release was increased by Ca2+, but was not completely blocked by EGTA. Simultaneous addition of 1 mM ATP with 1 mM MgCl2 (Mg-ATP) in the presence of Ca2+ reduced this release. However, in spite of the existence of Mg-ATP, the stimulation of PM-induced amylase release was caused by Ca2+ in a concentration-dependent manner (10(-7)-10(-3) M). These results suggest that Ca2+ and Mg-ATP should participate as important regulators in the exocytotic interaction between secretory granules and plasma membranes in this system. Furthermore, the differences between our system and intact cells are also discussed.

Adenosine Triphosphate

Time-dependent changes of collagen cross-links and their precursors in the culture of osteogenic cells.

The early stage of cross-link formation in bone collagen was studied in a cell culture system. An osteogenic cell line that produces and accumulates a remarkably high amount of collagen, and that eventually forms bone-like structures, was used in this study for its time-dependent development of reducible cross-links. It was found that precursors of the cross-link, dehydro-dihydroxynorleucine and dehydro-hydroxynorleucine became detectable as soon as the cells attained a confluent state. They showed maximal amounts at days 3-5 after confluence, but substantially disappeared at day 10 after confluence. In contrast, two characteristic cross-links of bone collagen, dehydro-dihydroxylysinorleucine (dehydro-DHLNL) and dehydro-hydroxylysinorleucine (dehydro-HLNL), which were present in trace amounts at the stage of cell confluence, gradually increased in amount and reached a plateau at day 10, just when their precursors disappeared. Thus, it was found that there was a time lag of about a week between the maximal formations of precursors and cross-links of bone collagen in this system. The significance of this time lag was interpreted in terms of the minimum essential accumulation of collagen for the precursor-product transition. The ratio of dehydro-DHLNL to dehydro-HLNL was as low as 0.7 at day 3 after confluency, increased to 4.2 at day 20, the period just before mineralization began, and decreased thereafter, suggesting a qualitative change in bone collagen associated with mineralization.

Animals

An osteonectin-like protein in the matrix of cultured osteogenic cell-line MC3T3-E1, which is associated with calcification.

Time-dependent changes of the [3H]-proline-labeled noncollagenous proteins synthesized by the osteogenic cell-line MC3T3-E1 were analyzed over a range starting from cell confluency to 13 days after confluency during which time cells formed a bone-like structure. It was found that the 40 kDa protein on SDS-polyacrylamide gel (SDS-PAGE) remarkably increased in the cell-matrix layer at about 9 days after cell confluence, just before calcification. This protein was highly purified and was found to contain high amounts of glutamic acid, glycine, and serine. An internal amino acid sequence of this protein was revealed to be K-X-M-A-P-E-E-X-P, which showed homology with the sequence of the EF-hand domain in osteonectin/SPARC (secreted protein, acidic, and rich in cysteine). This protein co-migrated with collagen in gel filtration, and ion-exchange chromatography. Furthermore, it showed high affinity to type I collagen.

Amino Acid Sequence

The natural course of osteoarthritis of the hip due to subluxation or acetabular dysplasia.

In 59 patients, 86 hips with subluxation or hip dysplasia were examined to determine the natural course of the condition and select suitable treatment. Thirty-three percent of the joints (13/39 hips) developed early osteoarthritis from pre-osteoarthritis within an average term of 9.2 years, while the remaining, sixty-six percent (31/47 hips) developed advanced-stage osteoarthritis from early osteoarthritis within an average term of 7.8 years. Patients were classified into advanced and non-advanced groups according to radiographic analysis of the advanced groups according to radiographic analysis of the advance of the disease and statistical analysis was performed. In pre-osteoarthritis, centre-edge angle, slope of the acetabular roof, acetabular head index, acetabular depth ratio and Japanese Orthopaedic Association (JOA) hip score were significant predictors, while in early osteoarthritis, a broken Shenton's line, cranial joint space and JOA score were significant. On the basis of multiple parameters, formulas for predicting development in patients with pre-osteoarthritis, those with early osteoarthritis, and all patients together were established, with an accuracy of 87%, 71%, and 68%, respectively.

Acetabulum

Hormonal regulation of the cell growth in an endometriotic cell culture system.

A primary cell culture of endometriotic cells was established to examine the hormonal regulation involved in the growth of endometriosis. The addition of estradiol had no effect on the DNA synthesis of cultured endometriotic cells. Progesterone at 10(-7) M caused a significant reduction in the amount of the DNA synthesis. An inhibitory effect of progesterone was completely eliminated by the concomitant addition of RU486, an antiprogesterone. The addition of epidermal growth factor (EGF) at 1 ng/ml significantly increased the DNA synthesis. Other growth factors, such as fibroblast growth factor and insulin-like growth factor also produced a modest but significant increase in the DNA synthesis. These results provide clues to the pathophysiology of endometriosis.

Cell Division

Hormonal regulation of endometriotic cell growth in primary cell culture system.

A primary cell culture of endometriotic cells was established to examine the hormonal regulation of endometriosis. The addition of estradiol had no effect on the DNA synthesis of cultured endometriotic cells. Progesterone at 10(-7) M caused a significant reduction in the amount of the DNA synthesis. An inhibitory effect of progesterone was completely eliminated by the concomitant addition of RU486, an antiprogesterone. The addition of epidermal growth factor (EGF) at 1 ng/ml significantly increased the DNA synthesis. Other growth factors, such as fibroblast growth factor and insulin-like growth factor also produced a modest but significant increase in DNA synthesis. These results seem to provide clues for clarifying the pathophysiology of endometriosis.

Cell Division

Hepatitis type C virus infection in patients with type B chronic liver disease.

Anti-c100-3 (Ortho) was determined in the sera of 152 patients with HBs antigen-positive chronic liver diseases to assess coinfection of hepatitis B virus (HBV) and hepatitis C virus (HCV). Eleven patients (7.2%) were positive for anti-c100-3. Anti-CP-9 (Okamoto) and HCV-RNA (RT-PCR) were also examined in these 11 patients. Anti-CP-9 was detected in 7 patients and HCV-RNA was detected in all 11 patients. Four of the 11 anti-c100-3-positive patients were positive for HBe antigen (HBeAg) and others were negative. In 8 of the 11 patients, HCV was suspected to be superinfected by blood transfusion. In HBeAg-positive patients, serum glutamic pyruvic transaminase (SGPT) was elevated in relation to active replication of HBV shown by DNA-polymerase activity. The histological findings showed chronic active hepatitis, with or without cirrhosis. On the other hand, in HBeAg-negative patients, SGPT fluctuated without evidence of active replication of HBV. Active inflammation in the liver was observed in 3 of 5 HBeAg-negative patients by liver biopsy. These findings suggest that HBV might play an important role in chronic active inflammation in HBeAg-positive patients coinfected with HCV, and that HCV might be responsible for continuous inflammation in HBeAg-negative patients coinfected with HCV.

Adult

Presence of a Vicia unijuga lectin-binding (Vgu) glycoprotein with Thomsen-Friedenreich (T) activity and Vgu glycoproteins in human primary hepatocellular carcinoma.

1. Twenty perchloric acid-soluble glycoprotein fractions (PASFs) were separated from carcinomatous and non-carcinomatous regions of the livers of 5 patients with primary hepatocellular carcinoma (HCC) and 5 patients with a complication of HCC and hepatocirrhosis (HC). 2. In autoradiography using 125I-labelled Vicia unijuga lectin (VUA) and 125I-labelled Arachis hypogaea anti-T lectin as probes, 10 PASFs from carcinomatous regions of the livers of 10 patients gave 1 Vgu glycoprotein with T activity and/or 1-4 Vgu glycoproteins, respectively, and among 10 PASFs from non-carcinomatous regions of the livers of 10 patients, 7 PASFs gave no glycoproteins, respectively. 3. These results show that a Vgu glycoprotein with T activity and Vgu glycoproteins occur in HCC and in a complication of HCC and HC as oncofetal antigens.

Antibodies

Two flavonol glycosides from Vancouveria hexandra.

In addition to two known glycosides, ikarisoside F and epimedin A, two new glycosides of a flavonol with a gamma, gamma-dimethylallyl group were isolated from the underground and the aerial parts of Vancouveria hexandra. The structures were determined to be des-O-methylanhydroicaritin 3,7-diglucoside and anhydroicaritin 3-glucosyl (1----3)rhamnoside-7-glucoside by means of spectral analysis.

Carbohydrate Sequence

Four flavonol glycosides from Achlys triphylla.

Four new flavonol glycosides were isolated from the underground parts of Achlys triphylla in addition to eight known compounds. By means of spectroscopic analysis, the structures were characterized as isorhamnetin 3-glucosyl(1----3)galactoside, isorhamnetin 3-[6''acetylglucosyl(1----3)galactoside], isorhamnetin 3-[4''6''-di-acetylglucosyl(1----3)galactoside], and syringetin 3-[6''-acetylglucosyl(1----3)galactoside], respectively. In the aerial parts of the plant, seven known compounds were also confirmed.

Carbohydrate Sequence

Flavonol glycoside production in callus cultures of Epimedium diphyllum.

Callus cultures of Epimedium diphyllum produced a large amount of epimedoside A in addition to a small amount of diphylloside B, ikarisoside C, epimedoside E, diglycosides of des-O-methylanhydroicaritin (8-gamma, gamma-dimethylallylkaempfero). Icariin, epimedins A-C, which are glycosides of anhydroicaritin, were also produced in the callus cultures. Contents of the flavonol glycosides in callus tissue were higher than those of mother plants, but the composition of each flavonol glycoside mixture in the callus cultures was different from that of the original plants. The time-course experiments showed that an inverse relationship existed between cell growth and flavonol glycoside production. Effects of hormonal factors on cell growth and flavonol glycoside production indicated that 2,4-dichlorophenoxyacetic acid was needed for the production of flavonol glycosides.

Carbohydrate Sequence

The use of multimedia in patient care.

A personal computer based system was constructed to assess the use of various forms of information (multimedia) in patient record keeping. A patient's file with his records kept in a multimedia fashion was made by using the system. We describe the hardware and software construction of the system together with the results and the memory requirements of each type of media. Potential usage of the system in the future is discussed especially in connection with the Picture Archiving and Communications Systems (PACS).

Medical Records Systems, Computerized

Biochemical and physiological changes induced by nicotinamide in a C3H mouse mammary carcinoma and CDF1 mice.

We have continued our investigation into the mechanism by which nicotinamide can enhance radiation damage in tumors, using a C3H mouse mammary carcinoma grown in CDF1 mice. Biochemical analysis of tumor extracts showed that nicotinamide (1000 mg/kg; i.p.) increased the ATP/Pi and ATP/ADP + AMP ratios. This change in metabolic activity was consistent with nicotinamide increasing tumor oxygenation. Moreover, the greatest effect occurred 0.5-2.5 hr after drug injection, a time at which radiosensitization by nicotinamide in this tumor had previously been shown to be maximal. These changes were observed without any apparent modification in tumor blood perfusion, measured using the 86-RbCl uptake procedure, and occurred despite nicotinamide producing a 50% decrease in mean arterial blood pressure, estimated directly by a carotid cannulation technique.

Adenosine Diphosphate

Comparison of cytokine levels and embryo toxicity in peritoneal fluid in infertile women with untreated or treated endometriosis.

OBJECTIVE: Our aim was to examine the relationship between the levels of cytokines in peritoneal fluid and its embryo toxicity. STUDY DESIGN: The levels of interleukin-1 and tumor necrosis factor were measured in peritoneal fluid from infertile women who did not have endometriosis (n = 21), who had untreated endometriosis (n = 19), and who had undergone medical treatment for endometriosis (n = 10). Embryo toxicity was investigated in mouse two-cell embryos cocultured with the oviducts in culture media that contained various concentrations of peritoneal fluid. RESULTS: The levels of cytokines were significantly higher in the peritoneal fluid from women who had untreated endometriosis than in women who did not have endometriosis, but they were extremely low in women who had undergone medical treatment with either danazol or buserelin. The peritoneal fluid from women who had untreated endometriosis adversely affected the cleavage of mouse two-cell embryos. After medical treatment the embryo toxicity of the peritoneal fluid was almost undetectable. CONCLUSION: These results offer some theoretic bases in support of medical treatment to improve reproductive performance in infertile women who have endometriosis.

Adult

[A case of HBsAg positive liver cirrhosis who died after withdrawal of steroid].

A 42-year-old male was admitted with subarachnoidal hemorrhage. Dexamethasone 224 mg was used to reduce brain edema. His operation was successful without blood transfusion. No remarkable signs and symptoms were found except HBsAg positive and mild GPT elevation during his admission. He was discharged on the 33rd day. But 2 weeks later, he felt general fatigue and became worse day by day. He was re-admitted on the 75th day. Several therapies were given but he died of hepatic failure on the 85th day. The autopsy showed liver cirrhosis with massive necrosis. We believed that the steroid-withdrawal-phenomenon caused excessive immunological response and this process caused his hepatic failure leading to death.

Adult