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Biomedical subjects

N Harada

Publications and source records attributed to N Harada.

At least 145 records · Page 8Linked to original sources
PubMed2000

Analysis of estrogen receptor alpha and beta in endometrial carcinomas: correlation with ER beta and clinicopathologic findings in 45 cases.

Estrogens play important roles in the pathogenesis of the great majority of endometrial endometrioid adenocarcinoma. Recently, a novel estrogen receptor (ER), ER beta, has been characterized, but little is known about the status of ER beta in endometrial carcinoma. We therefore examined expression of both ER alpha and ER beta in 45 cases of endometrioid endometrial adenocarcinoma using mRNA in situ hybridization, reverse transcription and polymerase chain reaction (RT-PCR), and immunohistochemistry. We also correlated the findings with various clinicopathologic parameters in these cases to examine their possible biologic significance. Accumulation of mRNA hybridization signals for both ER alpha and ER beta was detected predominantly in the cytoplasm of carcinoma cells, and to a lesser extent in some stromal cells. ER beta mRNA was detected in 16/45 cases (35.6%), and ER alpha mRNA hybridization signals were detected in 36/45 cases (80.0%). Among the 16 ER beta positive cases, 15 cases also had ER alpha mRNA hybridization signals. In the cases that expressed both ER alpha and ER beta, ER alpha mRNA hybridization signals were more widely distributed than ER beta mRNA. In 21 cases, carcinoma cells had ER alpha mRNA hybridization signals but not ER beta mRNA. There was a statistically significant positive correlation between the results of mRNA in situ hybridization and semiquantitative RT-PCR or immunohistochemistry for both ER alpha and ER beta. There were no significant correlations between ER beta mRNA expression and PR labeling index, Ki67 LI, age, or histologic grade. The results from our study indicate that ER beta is coexpressed with ER alpha, and that the estrogenic effects occur predominantly through ER alpha in endometrial carcinomas.

Adenocarcinoma↗
PubMed2000

The beneficial effect of mesalazine on esophageal ulcers in intestinal Behcet's disease.

Intestinal Behcet's disease in a 19-year-old girl was diagnosed because of the history of recurrent oral aphthous ulcers and typical endoscopic findings of esophageal and ileal ulcers. Her symptoms (e.g., dysphagia and retrosternal pain) were gradually relieved by treatment with prednisolone and total parenteral nutrition. However, about one month later, oral and esophageal ulcers appeared again. Mesalazine was added. Oral and esophageal ulcers healed promptly, and have not relapsed for about one year. Although mesalazine appears to act locally in the small intestine and colon, the therapeutic effect of mesalazine in this case may be explained by the systemic antiinflammatory effect. This case suggests that mesalazine is an effective drug and is a good candidate in the treatment of intestinal Behcet's disease, especially accompanied with esophageal involvement.

Adult↗
PubMed2000

Effect of acid-suppressive therapy on Helicobacter pylori production of interleukin-8 in the gastric mucosa.

BACKGROUND: Recent studies have shown that acid-suppressive therapy increases the severity of Helicobacter pylori- associated gastritis in the corpus. PURPOSE: To evaluate interleukin (IL)-8 production in the gastric corpus mucosa before and during acid-suppressive therapy in H pylori-infected patients. PATIENTS AND METHODS: Ten patients with reflux esophagitis (five H pylori-positive and five H pylori-negative) were treated with omeprazole 20 mg. Serum gastrin concentrations, H pylori colonization density and mucosal IL-8 levels in the corpus were investigated at entry and two weeks after starting therapy. IL-8 levels were measured by ELISA. The organism density was determined, and scored according to area occupied by the bacterial colonies. The presence of H pylori was assessed by rapid urease testing and histological finding of gastric biopsy specimens. RESULTS: In H pylori-positive patients, concentrations of IL-8 during therapy significantly exceeded those before therapy (36.2+/-6. 8 versus 18.3+/-3.8 pg/mg protein; P<0.05) without altering H pylori density. In H pylori-negative patients, IL-8 levels were similar before and during therapy (6.1+/-2.7 versus 6.3+/-3.0 pg/mg protein). Concentrations of gastrin during therapy were significantly higher than those before therapy in all patients. CONCLUSIONS: The results suggest that acid suppression increases mucosal IL-8 levels in H pylori-infected patients with reflux esophagitis.

Adult↗
PubMed2000

Down-regulation of CD98 in melphalan-resistant myeloma cells with reduced drug uptake.

Although melphalan has been used as a therapeutic reagent for multiple myeloma, many patients become refractory. To elucidate the mechanism of resistance to melphalan, we generated a melphalan-resistant myeloma cell line, KHM-11(EMS), by treating a parental line, KHM-11, with a mutagen, ethylmethanesulfonate. KHM-11(EMS) is 55 times more resistant to melphalan. gamma-Glutamylcysteine synthetase, P-glycoprotein, multidrug-resistance-associated protein, lung-resistance-related protein and the Bcl-2 family of proteins were not responsible for the drug resistance in KHM-11(EMS). Intracellular incorporation of melphalan to myeloma cells was determined by using [(14)C]-labeled melphalan. Accumulation of melphalan in KHM-11(EMS) was 43% of KHM-11, while the efflux rates were comparable in both cell lines. The uptake of melphalan was inhibited by the addition of L-phenylalanine, indicating that melphalan is incorporated through the L-phenylalanine transporter as reported previously. Expression of CD98, which was recently cloned as an L-phenylalanine transporter, was 6-fold decreased in KHM-11(EMS), suggesting that CD98 may be correlated with the incorporation of melphalan. CD98 expression and incorporation of melphalan were analyzed in fresh purified myeloma cells from 5 patients. All myeloma cells from 4 cases expressed CD98 at a high level and incorporated melphalan. However, tumor cells from 1 case expressed CD98 at low levels and did not incorporate melphalan. Taken together, reduced melphalan uptake could be responsible for the drug resistance in KHM-11(EMS), and down-regulation of CD98 may be related to this phenomenon. Further investigation of the correlation between impaired drug uptake and down-regulation of CD98 in myeloma cells should be important to understand the mechanism of resistance to melphalan.

ATP Binding Cassette Transporter, Subfamily B, Mem↗
PubMed2000

17beta-hydroxysteroid dehydrogenase type 1 and 2 expression in the human fetus.

The present study investigates the expression patterns of 17beta-hydroxysteroid dehydrogenase (17betaHSD) isozymes in human fetal tissues to understand how estrogenic activity is regulated in the human fetus. Using enzyme assay, high 17betaHSD activity was detected in the placenta and liver, and low levels of 17betaHSD activity were also present in the gastrointestinal tract and kidney. After Northern blot analysis, we detected the messenger ribonucleic acid for 17betaHSD type 1 (17betaHSD1) only in the placenta, whereas that for 17betaHSD type 2 (17betaHSD2) was detected in the placenta, liver, gastrointestinal tract, and urinary tract at 20 gestational weeks. In RT-PCR analysis of the messenger ribonucleic acid transcripts, 17betaHSD 1 was predominantly expressed in the placenta, brain, heart, lung, and adrenal, whereas 17betaHSD2 expression was predominantly detected in the liver, gastrointestinal tract, and kidney. In addition, we detected 17betaHSD2 immunoreactive protein in surface epithelial cells of the stomach, absorptive epithelial cells of the small intestine and colon, hepatocytes of the liver, and interstitial cells surrounding the urinary tubules of the renal medulla. 17betaHSD2 in these tissues may be functioning in the prevention of in utero exposure of the fetus to excessive estradiol from the maternal circulation and amniotic fluids.

17-Hydroxysteroid Dehydrogenases↗
PubMed2000

Enhancement of plant stem growth by flocculation of the antibiotic-producing bacterium, Pseudomonas fluorescens S272, on the roots.

The antibiotic-producing bacterium, Pseudomonas fluorescens, is assumed to be important in protecting plants from soilborne diseases. S. fluorescens S272, a hyper-producing strain of pyoluteorin (PT) and 2,4-diacetylphloroglucinol (DG), had previously been isolated from soil. The present paper reported that the growth of water-cultivated Kaiware radish was promoted to 120-140% of its normal level by the coaddition of an S272 culture broth (0.01-1% v/v) and a polysaccharide flocculant (1-100 ppm) from Klebsiella pneumoniae H12. Tight adhesion of S272 cells to the root tissue was microscopically observed. The growth promotion is assumed to have been caused by antibiotic effects for the following two reasons: 1) PT (4 mg/l) and DG (24 mg/l) addition to a radish culture enhanced stem growth to 130% of the normal level; 2) a culture solution containing the S272 culture broth (0.01-1% v/v) markedly inhibited the decomposition of hypersensitive chrysanthemum leaves. A soil-cultivation experiment with Gomphrena globosa under natural conditions also exhibited enhanced stem length (160%) by coaddition of the S272 culture broth and H12 polysaccharide. These results suggest that polysaccharide-enhanced adhesion of P. fluorescens S272 cells might be useful for promoting plant growth through the increased antibiotic effect.

Anti-Bacterial Agents↗
PubMed2000

Estrogen secreting adrenal adenocarcinoma in an 18-month-old boy: aromatase activity, protein expression, mRNA and utilization of gonadal type promoter.

We examined clinical, endocrinological and molecular biological aspects of an estrogen-secreting adrenal carcinoma in an 18-month-old male to clarify the pathogenesis of this condition. An 18-month-old boy was referred for evaluation of progressive bilateral gynecomastia and appearance of pubic hair. The patient had elevated plasma estradiol (349 pg/ml) and testosterone (260 ng/dl) levels that completely suppressed FSH and LH levels, and was subsequently diagnosed with an adrenal tumor on the right side. After removal of a 300-g adenocarcinoma, gynecomastia regressed and essentially normal hormone levels were restored. Aromatase activity in the tumor tissue determined by the 3H-water method was 71.0-104.4 pmol/min/mg protein. High levels of aromatase protein and mRNA in the tumor tissue were also demonstrated, while neither aromatase activity nor protein was detected in normal adrenal glands. To investigate the regulation of aromatase expression in the adrenal carcinoma, we examined the usage of alternate promoters responsible for aromatase gene transcription. In the present case, the amounts of aromatase mRNA utilizing gonadal types of exon 1c (1.3) and 1d (II) were significantly higher than those that using other exon 1s. This result suggested that the utilization of a gonadal-type exon 1 might be involved in the over-production of aromatase in estrogen-secreting adrenal carcinoma.

Adenocarcinoma↗
PubMed2000

[Is it able to simplify the classifications of staging and curability for pancreatic cancer?].

This study was undertaken to simplify the classifications of the staging and curability of pancreatic cancer by the Japan Pancreas Society (JPS). A total of 461 patients who underwent surgical resection from 1968 to 1997 were analyzed in this study in accordance with our new classification of staging and curability. This staging mainly consists of three factors based on tumor extent:invasion to the retroperitoneum (rp); invasion to the extrapancreatic nerve plexus (pl); and lymph node metastasis (n). Each factor is simply judged as + (positive) or - (negative). Curability is also judged simply based on whether the presence of tumor is detected or not, and the degree of lymph node dissection is not included in curability. Our staging and curability assessment is strongly associated with survival rates after surgery. The 5-year survival rate of stage I patients is 50% and curability of A is > or = 50%. It is concluded that it is possible to simplify the classification of the staging and curability of pancreatic cancer by the JPS.

Humans↗
PubMed2000

Synthesis, circular dichroism and absolute stereochemistry of 1,1',2,2',3,3',4,4',-octahydro-3,3'-dimethyl-4,4'-biphenanthryl

In the course of synthetic studies of chiral olefins, (3R,3'R)-(P,P)-(E)-(-)-1,1',2,2',3,3',4,4'-octahydro3,3'-dimethyl-4,4'-biphenanthrylidene (1) and its (3R, 3'R)-(P,P)-(Z)-(+)-isomer (2), chiral (3R,3'R,4R,4'R)-(-)-1,1',2,2',3,3',4,4'-octahydro-3,3'-dimethyl-4,4'-biphenanthryl (3) was obtained as a by-product in the McMurry reaction of (3R)-(-)-2,3-dihydro-3-methyl-4(1H)-phenanthrenone (4). The relative and absolute stereostructures of (-)-3 were fully determined by NMR, X-ray crystallographic, and CD spectral analyses.

Journal Article↗
PubMed2000

Circular dichroism and absolute stereochemistry of

Enantiopure [8]paracyclophane-10-carbonitrile (1) and related compounds were prepared. The absolute stereochemistry of [CD(+)242.0]-1 was determined to be R by theoretical calculation of its CD spectrum using the pi-electron SCF-CI-DV MO method. The theoretical determination came to the same conclusion as the X-ray crystallographic method had previously given. Other related compounds show similar CD spectra leading to the R absolute stereochemistry.

Journal Article↗
PubMed2000

Synthesis, circular dichroism, and absolute stereochemistry of a Fecht acid analog and related compounds

2,6-Dimethylspiro[3.3]heptane-2,6-dicarboxylic acid (2), an analog of the Fecht acid (1), was enantioresolved by the method of (1S,2R,4R)-(-)-2,10-camphorsultam yielding enantiopure acid (+)-2, the S absolute configuration of which was unambiguously determined by X-ray crystallography of camphorsultam amide derivative (R)-(-)-12b and chemical correlation. To determine the absolute configuration of chiral spiro[3.3]heptane compounds by the circular dichroism (CD) exciton chirality method, acid (+)-2 was converted to (+)-2,6-bis(phenylacetylenyl)-2,6-dimethylspiro[3.3]heptane (3) and (+)-2,6-bis(4-methoxyphenylacetylenyl)-2,6-dimethylspiro[3.3]heptane (4). The CD spectra of (+)-3 and (+)-4 exhibit intense exciton split Cotton effects of positive chirality, from which their S absolute configurations were confirmed.

Journal Article↗
PubMed2000

Adenosine and selective A(2A) receptor agonists reduce ischemia/reperfusion injury of rat liver mainly by inhibiting leukocyte activation.

To examine whether adenosine reduces ischemia/reperfusion (I/R)-induced liver injury by inhibiting leukocyte activation via A(2) receptor (A(2)R) stimulation, we investigated the effects of adenosine and selective A(2A) receptor (A(2A)R) agonists (YT-146 and CGS21680C) on I/R-induced liver injury in rats. Adenosine, YT-146, and CGS21680C, in the concentration of 10(-7) to 10(-5) M, significantly inhibited neutrophil elastase release by about 30 to 40% and increased intracellular Ca(2+) concentrations in isolated neutrophils stimulated with formyl-methionyl-leucyl-phenylalanine (fMLP) in vitro. Adenosine, YT-146, and CGS21680C, in the concentration of 10(-7) to 10(-5) M, significantly inhibited tumor necrosis factor (TNF)-alpha production by monocytes stimulated with endotoxin by about 50%. Although ZM241385, a selective A(2A)R antagonist, significantly enhanced the increase in neutrophil elastase release and intracellular Ca(2+) concentrations in neutrophils stimulated with fMLP, this agent did not affect the endotoxin-induced TNF-alpha production by monocytes. Rats were subjected to liver ischemia for 60 min. Serum levels of transaminases increased after hepatic I/R, peaking at 12 h after reperfusion. The i.v. infusion of adenosine (1 and 10 mg/kg/h), YT-146 (0.1 and 1 mg/kg/h), and CGS21680C (0.1 and 1 mg/kg/h) significantly inhibited the I/R-induced increase in serum transaminase levels 12 h after reperfusion. The I/R-induced decrease in hepatic tissue blood flow was significantly prevented by adenosine and YT-146. Hepatic levels of TNF-alpha, cytokine-induced neutrophil chemoattractant (equivalent to human interleukin-8), and myeloperoxidase were significantly increased after I/R. These increases were significantly inhibited by the administration of adenosine, YT-146, and CGS21680C. Although the histological neutrophil accumulation in the liver was significantly increased after I/R as evaluated by the naphthol AS-D chloroacetate technique, the administration of adenosine, YT-146, and CGS21680C significantly inhibited this increase. These findings suggest that adenosine reduces I/R-induced liver injury both by inhibiting the synthesis of inflammatory mediators and by inhibiting neutrophil degranulation directly, probably through A(2A)R stimulation.

Adenosine↗
PubMed2000

[Study of the two-step tuberculin tests and gamma-interferon assays in elderly persons].

Recently, an outbreak of lung tuberculosis among elderly residents happened in the home for aged, and it has become a social problem. To analyze the problem of tuberculosis in the elderly persons, we performed two-step tuberculin tests at two weeks interval and gamma-interferon assays for diagnosis of human tuberculosis to residents in the home for aged. 45 males (age; 77.3 +/- 8.9 SD) and 118 females (age; 81.9 +/- 8.6 SD) elderly persons (> or = 60) were examined. Standard dose of PPD was injected intracutaneously, and read at 48 hours. Erythema of 10 mm diameter or more was considered positive. The results were as follows. (1) The percentages of positive reactors in the 1st (T1) and the 2nd (T2) tuberculin tests were 50.3% and 68.1%, respectively. (2) The diameters of erythema and percentage of positive reactors (T1) did not very with age, but the size of erythema increased from 13.5 mm (T1) to 21.2 mm (T2) among those below 80 years, and from 13.0 mm (T1) to 16.4 mm (T2) among those 80 years or higher (p < 0.01). (3) The percentage of positive reactors (T1) varied by sex, namely males react stronger than females. (4) A dose of interferon-gamma released specifically in response to PPD stimulation of whole blood cultures showed no close relation with the diameters of erythema. In conclusion, tuberculin reactivity waned among elderly persons, and the fact suggests that they would have lower resistance against new infections, and as a result, they might have higher risk of developing tuberculosis. Elderly residents should be performed two-step tuberculin tests at the time of their entrance to the home for aged, and negative reactors should carefully be followed up.

Aged↗
PubMed2000

Comparison between short-term food restriction and exercise on whole body glucose disposal in high-fat rats.

High-fat diets induce whole-body insulin resistance. The aim of this study was to compare effect of two interventions: 3-day food restriction (66% of ad libitum fed) and 3-day exercise training (voluntary running wheels), on decreased insulin-mediated whole body glucose uptake in high-fat fed rats (5 mo old) using the hyperinsulinemic euglycemic clamp procedure. The control group was maintained on rat chow alone. After high-fat feeding for 2 wk, insulin-stimulated whole body glucose utilization was significantly decreased by 26%. The exercise training was more effective than food restriction in lowering plasma concentrations of insulin and triacylglycerol and tissue concentrations of triacylglycerol in soleus muscles. Diminished whole-body glucose uptake resulting from high-fat feeding was reversed completely by exercise training, but only partially by food restriction. The time course of starvation on insulin-stimulated glucose uptake was also observed in high-fat fed rats. Although the extension of starvation time to 48 h resulted in decreased plasma glucose, insulin and triacylglycerol concentrations, whole body glucose uptake did not increase further. These findings suggest that short-term exercise has a higher restorative effect on insulin sensitivity in high-fat fed rats than food restriction, in spite of the same loss in body weight, presumably due in part to improved local lipid availability.

Animals↗
PubMed2000

Cellular biological differences between human myeloma cell lines KMS-12-PE and KMS-12-BM established from a single patient.

To clarify cellular biological varieties of myeloma cells, biological differences were analyzed between 2 human myeloma cell lines, KMS-12-PE and KMS-12-BM, derived from pleural effusion and bone marrow, respectively, of a single patient. Although both lines were considered to be derived from the same clone because both had the same chromosomal marker and immunoglobulin H rearrangement, several biological differences were noted. CD11a and CD20 were highly expressed in the KMS-12-BM line, whereas the KMS-12-PE line showed a higher expression of CD7 and CD95/Fas. Although growth was stimulated in KMS-12-BM by interleukin-6 and interferon-alpha, it was inhibited in KMS-12-PE. In addition, apoptosis inhibitors Bcl-2 and Bcl-X(L) were highly expressed in KMS-12-BM cells. Because KMS-12-PE was cultivated 2 months before KMS-12-BM, these differences might be related to their origin (pleural effusion and bone marrow) or the phases of disease progression. However, these biological differences may help clarify myeloma cell biology and lead to improvement in treatment for myeloma patients.

Antigens, Surface↗
PubMed2000

The invasion of Mycobacterium tuberculosis into non-phagocytic cells.

To explore the ability of tubercle bacilli to invade and survive within non-phagocytic cells, we used in this study a human fibroblast cell line, WI-38, derived from normal embryonic lung and a human epithelial cell line, SQ-5, derived from lung squamous cell carcinoma. Live M. tuberculosis Erdman and M. tuberculosis H37Rv invaded WI-38 cells more efficiently than live M. tuberculosis H37Ra, M. bovis Ravenel, M. bovis BCG Tokyo and M. bovis BCG Pasteur. The capability of tubercle bacilli to invade WI-38 cells was Erdman > or = H37Rv > BCG Pasteur [symbol: see text] M. bovis Ravenel [symbol: see text] BCG Tokyo > H37Ra. A similar invasive ability was observed using SQ-5 cells. In contrast with live bacilli, heat-killed bacilli failed to invade WI-38 cells, whereas they were detected within SQ-5 cells. These results and incorporation of latex beads suggest that SQ-5 cells, but not WI-38 cells, possess phagocytic activity. H37Rv multiplied most actively within WI-38 cells when compared to H37Ra and BCG Tokyo, suggesting that the ability to invade and survive within non-phagocytic cells reflects the more active invasion of virulent M. tuberculosis than avirulent M. tuberculosis. The assay system used in this study may help us to clarify the virulence of tubercle bacilli in vitro.

Animals↗