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Biomedical subjects

H Ishida

Publications and source records attributed to H Ishida.

At least 631 records · Page 35Linked to original sources

[Effect of medroxyprogesterone acetate on the anticellular activity of 5-fluorouracil against human breast and stomach cancer cells].

As 5-fluorouracil (5-FU) has been known to show clinically antitumor effects against both breast and stomach carcinomas. We compared the combined effects of medroxyprogesterone acetate (MPA) with 5-FU on the growth of cultured human breast and stomach carcinoma cells. MPA inhibited the growth of estrogen-dependent human breast carcinoma MCF-7 cells at the low concentrations and exhibited an additive effect in combination with 5-FU. MPA also inhibited the growth of human stomach carcinoma MKN-45 cells at relatively high concentrations and exhibited an additive effect in combination with 5-FU. Human stomach carcinoma MKN-28 cells were rather insensitive to MPA, but, an additive combination effect of MPA and 5-FU was observed. These three cell lines were found to have MPA-binding proteins which may be distinct from the nuclear progesterone receptor, suggesting the correlation with growth-inhibitory activity of MPA.

Breast Neoplasms↗

[Effect of medroxyprogesterone acetate on antitumor efficacies and side effect of 5-fluorouracil].

Effect of medroxyprogesterone acetate (MPA) on the lethal toxicity, bone marrow toxicity and antitumor efficacies of 5-fluorouracil (5-FU) was examined. The following results were obtained. In aged female ddY mice, the lethal toxicity and body weight loss caused by 5-FU were reduced by MPA. MPA also reduced the bone marrow toxicity of 5-FU in aged female ddY mice. These effects of MPA were not observed in young male ddY mice. Moreover, MPA did not affect the antitumor activity of 5-FU against MPA-insensitive mouse breast carcinoma FM3A, or rather augmented its antitumor activity against MPA-sensitive human breast carcinoma MCF-7. These results indicate that MPA reduces the side effects of 5-FU and augments the antitumor activity of 5-FU in female mice.

Animals↗

[Diagnosis of mediastinal neoplasms using mediastinoscopy and CT-guided needle biopsy supported by gene analysis].

Of 140 cases of mediastinal neoplasms in our hospital, histological diagnosis was confirmed in 129 cases. We examined the methods of preoperative biopsy with those 129 cases. Biopsy had been performed in 25 cases. Mediastinoscopy was performed in seven cases, needle biopsy in eight cases, lymph node biopsy in eight cases, esophageal biopsy using a gastrofiberscope in one case, transbronchial biopsy using a bronchoscope in one case. The true positive rates of those methods were 100% for both mediastinoscopy and lymph node biopsy, and 75% for needle biopsy. Preoperative misdiagnosis occurred in two cases of needle biopsy. The postoperative histological diagnosis was malignant lymphoma in both cases. We performed gene analysis of the immunoglobulin heavy chain gene, light chain kappa and lambda genes, and the T-cell receptor beta gene by use of biopsied specimens, and we found rearrangement bands of these genes in the cases of malignant lymphoma. Therefore, we summarize that gene analysis is a reliable method if malignant lymphoma is suspected. If a needle biopsy is performed under CT guidance, the needle is sure to puncture the tumor. We concluded, therefore, that if a tumor is located in the anterior mediastinum, CT-guided needle biopsy should be performed first of all. Mediastinoscopy is a useful method if the tumor is located in the mid-mediastinum.

Biopsy, Needle↗

[An autopsy case of primary pulmonary malignant lymphoma associated with right pulmonary infiltrative shadow and macroglobulinemia].

We experienced a patient with two peaks of elevated monoclonal IgM and IgA in the serum, and lambda type Bence-Jones protein in the urine. Chest X-ray films showed right pulmonary infiltrative shadow and effusion. The results of the autopsy of this patient revealed infiltration of malignant lymphoma cells in both lungs, but in no other organs. The results of immunohistochemical staining for L26. UCHL-1, IgM, IgA and lambda chain in this case led to the diagnosis of diffuse small cell type. B-cell lambda, IgM type malignant lymphoma. Gene analysis using bone marrow did not reveal any rearrangement bands for the heavy chain, light chain kappa and lambda, or TcR beta genes. Therefore, we concluded that malignant lymphoma cells had infiltrated only the bilateral lungs, causing a rare case of primary pulmonary malignant lymphoma with two monoclonal peaks of increased serum IgM and IgA.

Aged↗

Clonal dental pulp cells (RDP4-1, RPC-C2A) synthesize and secrete osteopontin (SPP1, 2ar).

Dental pulp cells play an important role in maintaining dental mineralized tissue throughout life. Supplementary mineralization such as reparative dentin and pulp stone frequently occurs after primary dentin formation. Dental pulp cells are thought to be closely associated with such mineralization. We found that clonal rat dental pulp cells, RDP4-1 and RPC-C2A, produce and secrete osteopontin, but do not synthesize phosphophoryn which is a major noncollagenous protein found in dentin. The dental pulp osteopontin was highly phosphorylated and identified by thrombin susceptibility and immunoprecipitation with osteopontin/2ar antibody. Osteopontin synthesis markedly increased by 12-O-tetradecanoylphorbol-13-acetate (TPA) as observed in many osteoblastic cells. This study indicates that these cells can produce osteopontin as a major phosphoprotein and suggests that the synthesis of osteopontin could be used as a characteristic marker of dental pulp cells.

Animals↗

Purification and characterization of 7 alpha-hydroxy-4-cholesten-3-one 12 alpha-hydroxylase.

The isoform of cytochrome P450 that catalyzes the 12 alpha-hydroxylation of 7 alpha-hydroxy-4-cholesten-3-one, an intermediate in the conversion of cholesterol to cholic acid, was purified to homogeneity from rabbit liver microsomes. The extent of purification in the various steps was judged by an assay involving high performance liquid chromatography. The purified enzyme showed a single band on SDS-polyacrylamide gel electrophoresis (M(r) = 50,000). The NH2-terminal amino acid sequence is as follows: Val-Leu-Trp-Gly-Leu-Leu-Gly-Ala-Leu-Leu-Met-Val-Met-Val-Gly-, which is different from that of any other P450s so far reported. The specific content of the enzyme was 13.3 nmol of cytochrome P450/mg of protein. Upon reconstitution with NADPH-cytochrome P450 reductase and cytochrome b5, the P450 enzyme showed a high activity of 12 alpha-hydroxylation with a turnover number of 36.6 min-1 at 37 degrees C. The omission of either cytochrome P450 or NADPH-cytochrome P450 reductase resulted in complete loss of activity, and the omission of cytochrome b5 resulted in 40% loss of activity. Antibodies prepared from mouse inhibited the 12 alpha-hydroxylase activity of rabbit liver microsomes about 90% and that of the rat liver microsomes 50%. The enzyme activity was not inhibited by other antibodies raised against other forms of P450 that catalyze different monooxygenation reactions toward xenobiotics or endogenous substrates. Anti-cytochrome b5 antibody inhibited the activity 40%, suggesting the functional role of this protein, and anti-reductase inhibited the activity almost completely. The microsomal enzyme activity was markedly elevated by starvation or streptozotocin administration to the animals. However, an immunoblotting experiment showed no correlation between the enzyme activity and the amount of protein, suggesting that post-translational modification may occur.

Amino Acid Sequence↗

Bone marrow transplantation as a strategy for treatment of non-insulin-dependent diabetes mellitus in KK-Ay mice.

The effects of allogeneic bone marrow transplantation (BMT) on non-insulin-dependent diabetes mellitus (NIDDM) were examined using KK-Ay mice. KK-Ay mice reconstituted with KK-Ay bone marrow cells showed glycosuria, hyperinsulinemia, and hyperlipidemia. However, KK-Ay mice (H-2b) that had been lethally irradiated (9.0 Gy) and then reconstituted with T cell-depleted bone marrow cells from normal BALB/c mice (H-2d) showed negative urine sugar with decreases in serum insulin and lipid levels 4 mo after BMT. Morphological recovery of islets and glomeruli was also noted after allogeneic BMT. These findings suggest that BMT can be used to treat not only a certain type of NIDDM but also its complications such as hyperlipidemia and diabetic nephropathy.

Animals↗

Group II phospholipase A2 induced by interleukin-1 beta in cultured rat gingival fibroblasts.

Previously, we reported the presence of group II-like phospholipase A2 activity in the soluble fraction of rat gingiva. In the present study, we found that treatment of rat gingival cells with human recombinant interleukin-1 beta resulted in dose-dependent stimulation of intracellular and extracellular phospholipase A2 activity. Antisera against group II phospholipase A2 totally blocked the interleukin-1 beta-induced phospholipase A2 activity, but antisera against group I phospholipase A2 did not. Moreover, immunoblot analysis showed that the induced phospholipase A2 was group II phospholipase A2. These findings suggest that the induced enzyme belongs to the group II phospholipase A2 family of proinflammatory enzymes.

Animals↗

Continuous anti-interleukin 10 antibody administration depletes mice of Ly-1 B cells but not conventional B cells.

Ly-1 B cells have the distinctive property of continuous self-replenishment and, as we have shown previously, can be further distinguished from conventional B cells on the basis of greatly elevated constitutive and inducible production of the recently described cytokine interleukin 10 (IL-10). To test the possibility that IL-10 acts as either an autocrine or paracrine growth factor for Ly-1 B cells, we treated mice continuously from birth to 8 wk of age with a monoclonal rat IgM antibody that specifically neutralizes mouse IL-10. Mice treated in this way lacked peritoneal-resident Ly-1 B cells, contained greatly reduced serum immunoglobulin M levels, and were unable to generate significant in vivo antibody responses to intraperitoneal injections of alpha 1,3-dextran or phosphorylcholine, antigens for which specific B cells reside in the Ly-1 B cell subset. In contrast, conventional splenic B cells of anti-IL-10-treated mice were normal with respect to total numbers, phenotype, and in vitro responsiveness to B cell mitogens and the thymus-dependent antigen trinitrophenyl-keyhole limpet hemocyanin (TNP-KLH). The mechanism of Ly-1 B cell depletion appeared to be related to elevation of endogenous interferon gamma (IFN-gamma) levels in anti-IL-10-treated mice, since coadministration of neutralizing anti-IFN-gamma antibodies substantially restored the number of peritoneal-resident Ly-1 B cells in these mice. These results implicate IL-10 as a regulator of Ly-1 B cell development, and identify a procedure to specifically deplete Ly-1 B cells, thereby allowing further evaluation of the role of these cells in the immune system.

Animals↗

Synthesis of an S-(alpha-sialosyl)-(2----9)-O-(alpha-sialosyl)-(2----3')-beta-lactos ylceramide.

A ganglioside GD3 analog has been synthesized having an N-acetylneuraminic acid (Neu5Ac) residue alpha-thioglycosidically linked to C-9 of the Neu5Ac residue in the ganglioside GM3 structure. Glycosylation of 2-(trimethylsilyl)ethyl O-(6-O-benzoyl-beta-D-galactopyranosyl)-(1----4)-2,6-di-O-benzoyl-beta-D -glucopyranoside with methyl (methyl 5-acetamido-4,7,8-tri-O-acetyl-9-bromo-3,5,9-trideoxy-D-glycero-alpha-D- galacto-2-nonulopyranosid)onate, which was prepared from methyl (methyl 5-acetamido-3,5-dideoxy-D-glycero-alpha-D-galacto-2- nonulopyranosid)onate by 8,9-O-isopropylidenation, O-acetylation, hydrolysis of the isopropylidene group, selective bromination, and O-acetylation, using dimethyl(methylthio)sulfonium triflate (DMTST) as a promoter, gave the alpha-sialosyl-(2----3')-lactoside 8. Coupling of the O-acetyl derivative of 8 with the sodium salt of methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-3,5-dideoxy-2-thio-D-glycero-alpha-D- galacto-2-nonulopyranosonate gave an alpha-thioglycosidically linked tetrasaccharide. This was converted, via selective removal of the 2-(trimethylsilyl)ethyl group, trichloroacetimidation, and glycosidation with (2S,3R,4E)-2-azido-3-O-benzoyl-4-octadecene-1,3-diol into a ganglioside precursor. Finally the precursor on selective reduction of the azide group, coupling with octadecanoic acid, O-deacylation, and de-esterification gave the analog, S-(N-acetyl-alpha-neuraminosyl)-(2----9)-O-(N-acetyl-9-thio-alpha- neuraminosyl)-(2----3')-beta-lactosylceramide.

Antigens, CD↗

The role of 5'-methylthioadenosine on rat calvaria cell differentiation.

The role of 5'-methylthioadenosine (MTA), formed during the process of polyamine biosynthesis, on differentiation of osteoprogenitor cells was assessed by its effects on alkaline phosphatase (ALP) activity, bone nodule formation and osteopontin contents of cultured rat calvaria (RC) cells. These three markers were stimulated by exogenous MTA and were depressed by 5'-difluoromethylthioadenosine (DFMTA), a synthetic inhibitor of MTA phosphorylase, which cleaves MTA to adenine and 5-methylthioribose-1-phosphate. 5-Methylthioribose and 2-keto-4-methylthiobutyrate, metabolites of 5-methylthioribose-1-phosphate, had no effects on ALP activity and bone nodule formation in the presence or absence of DFMTA. On the other hand, adenine enhanced ALP activity, bone nodule formation and osteopontin contents in mineralized nodules and also partially reversed DFMTA-induced inhibition of these three markers. MTA, its metabolites and DFMTA did not affect the growth of RC cells under these culture conditions. These results suggest that adenine formed from MTA is important in the differentiation of RC cells.

Adenine↗