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Biomedical subjects

M Okamura

Publications and source records attributed to M Okamura.

At least 55 records · Page 3Linked to original sources

Nitrite-specific active transport system of the cyanobacterium Synechococcus sp. strain PCC 7942.

Studies on the nitrite uptake capability of a mutant of Synechococcus sp. strain PCC 7942 lacking the ATP-binding cassette-type nitrate-nitrite-bispecific transporter revealed the occurrence of a nitrite-specific active transport system with an apparent Km (NO2-) of about 20 microM. Similar to the nitrate-nitrite-bispecific transporter, the nitrite-specific transporter was reversibly inhibited by ammonium in the medium.

Biological Transport, Active↗

Salivary amylase activity of rats fed a low calcium diet.

Wistar strain rats fed low calcium diets (1, 2, 3, 4, 6 and 8 weeks) exhibited changes in secretory function of whole saliva. In particular, there were changes in salivary flow rate, total salivary protein, amylase enzyme activity, salivary amylase content and the level of cyclic AMP in the parotid gland acinar cell. Although there were no changes for the first 3 weeks, the levels increased at week 4 and decreased at week 6. The wet weight of the parotid gland started to decrease at week 4. These results suggest that when fed low calcium diets for long periods of time, rats develop defective salivary secretion.

Amylases↗

Separative determination of ascorbic acid analogs contained in mushrooms by high-performance liquid chromatography.

Analogs (6-deoxyascorbic acid, erythroascorbic acid, and associated glycosides) of L-ascorbic acid (AA) contained in mushrooms were allowed to react with hydrazine to form osazones, and the conditions for separative determination by HPLC using a Zorbax SIL column were examined. Separation was started using solvent system 1 (ethylacetate/n-hexane/acetone/acetic acid, 50:50:1:1, v/v) as the mobile phase, and switching after 15 min to solvent system 2 (ethylacetate/acetone/acetic acid, 100:1:1, v/v). Detection was performed by absorbance at 500 nm. Because these analogs showed different formation rates for osazone, calibration curves were prepared for each substance. The recovery rate in the load test was 93-105%. By this method, AA and the analogs contained in eight species of edible mushrooms have been determined. The results revealed that: (1) the main constituents of all mushrooms are AA analogs rather than AA itself; (2) only one species contained AA in a very small amount (2 mumol/kg); (3) the types of AA analogs present differed according to the species of mushrooms, and (4) the total amount of AA analogs was between ca. 100-500 mumol/kg (2-9 mg per 100 g, converted to AA). In addition, a new AA analog was found in Pleurotus ostreatus and identified as 5-O-(alpha-D-xylopyranosyl)-D-erythroascorbic acid in structural analyses by NMR and other methods.

Ascorbic Acid↗

Involvement of hepatocyte growth factor in formation of bronchoalveolar structures in embryonic rat lung in primary culture.

To clarify the role of hepatocyte growth factor (HGF) in embryonic lung development, organoids from fetal rat lung were cultured in collagen gels with or without HGF antisense oligonucleotides. Cyst-like structures formed within 24 h in organoids isolated from fetuses after 14 days' gestation, but this was abolished by the oligonucleotide addition, apparently by interference with the endogenous expression of HGF. Electron microscopy revealed two types of structure: an alveolar type characterized by osmiophilic lamellar bodies in the cytoplasm and lumen, and a bronchial type consisting of epithelial cells bearing microvilli on their apical surfaces. HGF mRNA was detectable from day 14 in fetal lung by RT-PCR. Our results suggest that HGF plays, coordinately with its expression, a crucial role in the morphogenesis of both alveolar and bronchial epithelia in the rat fetal lung.

Animals↗

Fetal heart rate patterns associated with periventricular leukomalacia.

OBJECTIVE: Our purpose was to assess the applicability of fetal heart rate (FHR) monitoring to detect fetuses at risk of developing periventricular leukomalacia (PVL). METHODS: FHR tracings obtained for babies delivered under 33 weeks' gestation and with a birth weight under 2000 g were assessed for baseline heart rate, variability, deceleration and "flip flap' (an oscillatory tracing pattern). RESULTS: PVL developed in 19 of the 103 infants studied. All of these infants were among the fetuses who exhibited average and increased variability. In addition, PVL was detected in 10 (47.6%) of the 21 flip flap positive fetuses, and in 9 (11.0%) of the 82 flip flap negative fetuses. The incidence of PVL was significantly higher in the flip flap positive fetuses (P < 0.005). CONCLUSION: The possibility that an unstable intrauterine environment, reflected by a flip flap pattern, is related to the subsequent development of PVL is indicated.

Female↗

Anti-angiogenic compound (TNP-470) inhibits mesangial cell proliferation in vitro and in vivo.

Growth factors, especially basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and transforming growth factor-beta (TGF-beta) are known to play key roles in the pathogenesis of mesangial proliferative glomerulonephritis. TNP-470 (AGM-1470), a potent anti-angiogenic compound, has anti-growth factor properties and inhibits the activation of cyclin-dependent kinase (cdk) 2 and phosphorylation of RB protein. We investigated whether TNP-470 could suppress growth factor induced mesangial cell proliferation in vitro and experimental model of mesangial proliferative glomerulonephritis in vivo. TNP-470 inhibited potently PDGF- and bFGF-stimulated proliferation of rat mesangial cells in vitro (IC50 = 50 pg/ml). In anti-Thy 1.1 glomerulonephritis, high dose use of TNP-470 (20 mg/kg/day) markedly suppressed mesangial cell proliferation and mesangial matrix expansion on day 6; however, mesangiolysis remained. Low dose use of TNP-470 (10 mg/kg/day) moderately inhibited mesangial cell proliferation and mesangial matrix synthesis, and induced appropriate glomerular healing on day 14 in anti-Thy 1.1 glomerulonephritis. Thus, TNP-470 potently inhibits growth factor-induced proliferation of mesangial cells in vitro, and mesangial cell proliferation and extracellular matrix expansion in anti-Thy 1.1 glomerulonephritis in vivo. These results suggest a novel therapeutic potential of TNP-470 in mesangial proliferative glomerulonephritis.

Animals↗

Immunohistochemical localization of placental leucine aminopeptidase/oxytocinase in normal human placental, fetal and adult tissues.

While oxytocinase is known to exist in pregnancy serum and placenta, the present study describes the expression of the mRNA for this enzyme in a wide variety of other human tissues. Northern blot analysis was used to detect the mRNA, with a probe derived from a cDNA for oxytocinase/placental leucine aminopeptidase (P-LAP). Both the distribution and localization of immunoreactive oxytocinase/P-LAP protein have been determined immunohistochemically by use of an anti-P-LAP antibody in normal placental, fetal and adult tissues. In placental tissues, only syncytiotrophoblasts were stained positively. In both fetal and adult tissues, positive staining was obtained in vascular endothelial cells, gastrointestinal mucosal cells, epithelial cells of hepato-biliary, pancreato-biliary, bronchial-alveolar and renal tubular systems as well as islet cells of pancreas and neurons in the central nervous systems. Sweat-gland cells, seminal vesicles and prostate gland in the adult, as well as adipocytes and skeletal muscle cells in the fetus were also stained. The widespread distribution of P-LAP suggests its involvement in a variety of physiological events not restricted to the regulation of the amounts of bioactive peptides such as arginine vasopressin (AVP) and oxytocin (OT) in pregnancy. The presence of P-LAP in syncytiotrophoblasts supports the idea that P-LAP in pregnancy serum is derived from the placenta.

Adult↗

Enhanced gene expression of scavenger receptor in peripheral blood monocytes from patients on cuprophane haemodialysis.

BACKGROUND: Macrophage scavenger receptor (SR) is implicated in playing a key role in macrophage-derived foam cell formation by taking up a large amount of modified low-density lipoproteins (LDL). It has also been postulated that alpha 2-macroglobulin receptor/LDL receptor-related protein (alpha 2MG/LRP) is involved in the development of foam cells by taking up apo E-enriched chylomicrons and VLDL remnants, and lipoprotein lipase-triglyceride-rich lipoprotein complexes. Accumulation of these lipid-loaded monocyte/ macrophages in the subendothelial space is considered to be an early event of atherogenesis. Since atherogenesis is considered to be accelerated in dialysis patients, we attempted to investigate whether gene expression of SR and alpha 2MG/LRP are altered in peripheral blood monocytes from patients on haemodialysis with a cuprophane (Cu) or polymethylmethacrylate (PMMA) membranes. METHODS: Peripheral blood monocytes (PBM) were prepared from patients undergoing haemodialysis with a Cu membrane (n = 9), patients undergoing haemodialysis with a PMMA membrane (n = 9), and healthy controls (n = 7). In a separate experiment we examined SR gene expression in uraemic patients (n = 12) and healthy controls (n = 9). SR and alpha 2MG/LRP mRNA were semiquantitated using reverse-transcription polymerase chain reaction (RT-PCR) assay followed by Southern blotting. RESULTS: SR mRNA expression in PBM from patients on chronic haemodialysis with a Cu membrane was about twofold higher than that in PBM from patients on chronic haemodialysis with a PMMA membrane or the controls (P < 0.05). alpha 2MG/LRP mRNA expression in PBM showed no difference among these, three groups. SR gene expression in monocytes from uraemic patients was not increased compared with that in the controls. CONCLUSION: PBM from patients under Cu membrane dialysis showed higher gene expression of SR than patients under PMMA membrane dialysis, uraemic patients, or healthy controls. This increased gene expression of SR in monocytes may be associated with the pathogenesis of accelerated atherosclerosis in patients on dialysis with a Cu membrane.

Arteriosclerosis↗

Favorable response to heparin in a pregnant woman with possible glomerular thrombosis as a complication of systemic lupus erythematosus and anti-phospholipid antibody syndrome.

A rare case of possible glomerular thrombosis during pregnancy is reported in a patient with active systemic lupus erythematosus and the presence of anti-phospholipid antibodies. Acute renal impairment was restored by administering infusions of heparin. Cesarean section was performed due to fetal distress, and resulted the live birth of a healthy infant.

Adult↗

Bulge ductility of several occlusal contact measuring paper-based and plastic-based sheets.

Articulating paper/film is a sheet strip that is coated with ink- or dye-containing wax and is used for marking occlusal contacts and monitoring the results. New types of occlusal film systems have been developed recently, which are capable of being computer-interfaced to identify the occlusal contact points and digitize the occlusal force. The mechanical ductility and thickness of these occlusal sheets constitute some of their important properties. The objective of this study was to evaluate seven different occlusal sheets and compare their bulge ductility and thickness. A custom-designed photo-sensing bulge tester was used. Three paper-based sheets (BAP, BET and SFA), two plastic-based sheets (ACF and AOS), and advanced systems (TSS and FDP) were tested. The specimen size was 20 mm square. Fifteen samples were tested for each material. The sheet film thickness with coated layers was as follows: BAP, 62 microns; BET, 46 microns; SFA, 133 microns; ACF, 23 microns; AOS, 14 microns; TSS, 134 microns; and FDP, 82 microns. Ductility (standard deviation) was as follows: BAP, 2.10 (0.0060)%; BET, 2.14 (0.008)%; SFA, 5.19 (0.57)%; ACF, 8.68 (0.05)%; AOS, 16.26 (0.41)%; TSS, 16.26 (0.41)%; and FDP, 6.37 (0.09)%. One-way ANOVA analysis indicated that (1) there in no statistical difference between BAP and BET (p > 0.001), (2) there is a statistical difference (p < 0.001) among all the rest of the tested occlusal sheets, and (3) bulge ductility appears to be linearly related to film thickness, its correlation depending upon types of base materials.

Coloring Agents↗

Lovastatin inhibits gene expression of type-I scavenger receptor in THP-1 human macrophages.

Lovastatin, an inhibitor of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase, inhibits the synthesis of mevalonic acid and is widely used as an anti-atherosclerotic drug. The macrophage scavenger receptor (SCR), a trimeric membrane glycoprotein, is postulated to play a key role in atheroma macrophage foam cell formation. HMG-CoA reductase is involved in the control of the synthesis of glycoproteins and farnesylated proteins, including ras proteins, which are involved in the transcriptional regulation of SCR gene expression. Accordingly, we examined whether lovastatin alters the gene expression of SCRs in THP-1 cell derived human macrophages. Lovastatin (5-15 microM) caused a significant dose-related reduction in steady state levels of type-I SCR mRNA in phorbol 12-myristate 13-acetate (PMA)-treated THP-1 cells. The addition of exogenous mevalonate (1 mM) completely restored the lovastatin-induced decrease of type-I SCR mRNA levels. While the addition of the isoprenoid end-product, isopentenyl adenine (50 microM), had little effect on the type-I SCR mRNA levels in lovastatin treated cells, the addition of isoprenoid farnesol (5 microM) largely restored the lovastatin-induced decrease of type-I SCR mRNA levels. Actinomycin D treatment showed that degradation rates of type-I SCR mRNA did not differ between the THP-1 derived cells with and without lovastatin treatment. Nuclear run-on assays showed that lovastatin markedly decreased the transcription of SCR gene in the cells. These results suggest that lovastatin inhibits the transcription of type-I SCR gene by affecting mevalonate metabolism, possibly through the farnesyl-pyrophosphate related end-product(s) in the THP-1-derived macrophages.

Cell Line↗

Ligand recognition in mu opioid receptor: experimentally based modeling of mu opioid receptor binding sites and their testing by ligand docking.

For three-dimensional understanding of the mechanisms that control potency and selectivity of the ligand binding at the atomic level, we have analysed opioid receptor-ligand interaction based on the receptor's 3D model. As a first step, we have constructed molecular models for the multiple opioid receptor subtypes using bacteriorhodopsin as a template. The S-activated dihydromorphine derivatives should serve as powerful tools in mapping the three-dimensional structure of the mu opioid receptor, including the nature of the agonist-mediated conformational change that permits G protein-coupling to "second messenger' effector molecules, and in identifying specific ligand-binding contacts with the mu opioid receptor. The analyses of the interactions of some opioid ligands with the predicted ligand binding sites are consistent with the results of the affinity labeling experiments.

Affinity Labels↗

Immunohistochemical and enzyme histochemical characteristics of short axon cells in the olfactory bulb of the golden hamster.

Short axon (SA) cells in the olfactory bulb are subdivided into six types after Golgi impregnation, although their functional significance is not fully elucidated. In the present study, we examined the golden hamster olfactory bulb by immunohistochemistry to localize neurotransmitters, neuron-specific marker, and nitric oxide synthase (NOS) in the SA cells. Enzyme histochemical staining was also performed to detect the activity of nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase, which is identified with NOS. In the main olfactory bulb (MOB), neuropeptide Y (NPY)-, NOS-, and NADPH-diaphorase-positive SA cells were detected in the glomerular layer (GL), vasoactive intestinal polypeptide (VIP)-positive SA cells in the external plexiform layer (EPL), and NPY-, somatostatin (SOM)-, protein gene product 9.5 (PGP 9.5)-, NOS-, and NADPH-diaphorase-positive SA cells in the granule cell layer (GCL). In the accessory olfactory bulb (AOB), VIP- and PGP 9.5-positive SA cells were detected in the mitral/tufted cell layer (MTL), and NPY-, SOM-, NOS-, and NADPH-diaphorase-positive SA cells in the GCL. The common presence of NPY- SOM-, VIP-, PGP 9.5-, NOS-, and NADPH-diaphorase-positive SA cells in both the MOB and the AOB may suggest that respective types of cells with the same immunoreactivity play the same role no matter where these cells are located in the MOB or the AOB.

Animals↗

Adult respiratory distress syndrome induced by a Chinese medicine, Kamisyoyo-san.

We report the first case of Kamisyoyo-san-induced adult respiratory distress syndrome (ARDS). A 59-year-old female was given Kamisyoyo-san for treatment of seborrheic dermatitis. She then presented with a respiratory illness having clinical, radiologic and functional characteristics of ARDS. Bronchoalveolar lavage fluid showed an increased number of lymphocytes, neutrophils and eosinophils. The lymphocyte stimulation test with Kamisyoyo-san was positive.

Bupleurum↗

Comparison of three methods for analysis of urinary sediments.

Efficiency of urine sediment analysis was evaluated in 423 specimens including 86 documented renal disorder ones using three methods; a traditional centrifugal and microscopic one, an image-scanning one and a flow cytometric one. Among the three methods, good correlations were observed in white blood cells and epithelial cells. Between white blood cells and bacterial counts a good correlation was observed only in the flow cytometry one. Female specimens usually demonstrated high values in white blood cells except with the microscopic method. Casts in patients with renal disorder exhibited significantly high in the flow cytometric method. In all methods, proteinuria specimens showed high cast count in the sediment. From these results we concluded that the flow cytometric method was clinically useful, especially in urinary tract disorder specimens.

Adolescent↗

Urinary excretion of type IV collagen and laminin in the evaluation of nephropathy in NIDDM: comparison with urinary albumin and markers of tubular dysfunction and/or damage.

To clarify the diagnostic relevance of urinary type IV collagen (IV-C) and laminin in diabetic nephropathy, the excretion of these basement membrane proteins were determined by enzyme immunoassay in 172 non-insulin-dependent diabetic patients with different grades of nephropathy and 64 non-diabetic control subjects, and were evaluated in comparison with those of urinary albumin, N-acetyl-beta-D-glucosaminidase (NAG) and alpha 1-microglobulin (alpha 1MG). These excretions were also compared between a group of non-diabetic renal disease (NDRD) patients (n = 24) and a subgroup of the diabetic patients studied (n = 76), whose urinary albumin excretion (UAE) varied within the ranges of micro- and macroalbuminuria. Of the diabetic patients studied, 49.7%, 53.4% and 32.4% had raised urinary albumin, NAG and alpha 1 MG excretion, respectively. In these patients, 54% and 53% exceeded the upper limit of normal for urinary IV-C and laminin. The level of IV-C and laminin excretion and the prevalence of their abnormal excretion showed a trend to increase with increasing grade of nephropathy, as assessed by UAE. In the normoalbuminuric [UAE < 20 mg/g creatinine (Cr)] stage, 28.3% and 26.3% patients had raised urinary IV-C and laminin excretion, respectively. In this stage, the excretion values for IV-C and laminin also rose significantly even when the UAE was < or = 10 mg/g Cr (P < 0.05 and P < 0.005, respectively). There was a close linear relationship between IV-C and laminin excretion (r = 0.73, P < 0.0001), together with their significant relationships with albumin, NAG and alpha 1MG excretion. The relationship of urinary IV-C and laminin with urinary NAG and alpha 1MG excretion remained significant even in normoalbuminuric patients. The normoalbuminuric patients with raised NAG and/or alpha 1MG excretion also had a higher prevalence of raised IV-C and laminin excretion than those with normal NAG and alpha 1MG excretion. The excretion values for IV-C and laminin, and the excretion ratios for IV-C/albumin and laminin/albumin were significantly higher in diabetic patients with evidence of incipient and clinical nephropathy than in NDRD patients, though the two patient groups had a comparable level of serum Cr and UAE. We conclude that the measurement of urinary IV-C and laminin may have potential for the evaluation of diabetic nephropathy. Furthermore, their determination might be helpful for distinguishing diabetic versus non-diabetic etiologies of altered renal function in diabetic patients.

Acetylglucosaminidase↗

Blood pressure regulates platelet-derived growth factor A-chain gene expression in vascular smooth muscle cells in vivo. An autocrine mechanism promoting hypertensive vascular hypertrophy.

To clarify the role of PDGF A-chain in hypertensive vascular hypertrophy of spontaneously hypertensive rats (SHRs), we studied levels of PDGF A-chain gene expression and transcription factors related to the gene in vascular smooth muscle cells (VSMCs) of SHRs in vivo. RNase protection assay and in situ hybridization showed that PDGF A-chain mRNA levels in VSMCs of SHRs were twofold higher than in those of normotensive Wistar-Kyoto rats. Gel retardation assays showed that levels of Sp1 and AP-2 in VSMCs of SHRs were twofold more abundant than in those of Wistar-Kyoto rats. Treatment with four pharmacologically different species of antihypertensive drugs for 2 wk decreased the levels of both PDGF A-chain mRNA and Sp1, but not AP-2 level in VSMCs of SHRs with regression of aortic hypertrophy, indicating that increases in levels of both PDGF A-chain mRNA and Sp1 in VSMCs of SHRs were associated with high blood pressure. These results suggest that high blood pressure is a stimulus which upregulates PDGF A-chain gene expression in VSMCs of SHRs, resulting in an autocrine enhancement in hypertensive vascular hypertrophy, and that the activation of the gene may be mediated through increases in Sp1 in these cells.

Animals↗

Retroperitoneal approach for lumbar lateral meningocele--case report.

A 29-year-old female with neurofibromatosis presented with a right lumbar lateral meningocele. Abdominal computed tomography (CT) showed a huge right retroperitoneal cyst expanding anterolaterally and displacing the right kidney. CT following myelography disclosed the cyst expanding through a wide defect of the right pedicles of the T-12 and L-1 vertebrae. The cyst was resected through a retroperitoneal approach with right flank oblique incision. Postoperatively, cerebrospinal fluid leakage occurred, which improved after lumboperitoneal shunt. Careful screening for lateral meningocele, including the lumbar region, should be undertaken in a patient with neurofibromatosis who presents with vertebral anomalies.

Adult↗