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Biomedical subjects

K Nagashima

Publications and source records attributed to K Nagashima.

At least 235 records · Page 13Linked to original sources

Expression of vascular endothelial growth factor in human myocardial infarction.

To define mechanisms that may influence collateral circulation and angiogenesis, we investigated vascular endothelial growth factor (VEGF) mRNA expression in human hearts. In non-ischemic human hearts, VEGF mRNA was not detected in vessels, but was found in cardiomyocytes. In hearts with myocardial infarction, the intensity of the VEGF signal was much higher in smooth muscle cells of arterioles adjacent to necrosis and in infiltrating macrophages than in myocytes around the site of the necrosis. This study suggests that levels of VEGF expression are high in smooth muscle cells and macrophages around infarcted areas after myocardial infarction and that VEGF may play a role in promoting collateral circulation and angiogenesis in human ischemic hearts.

Aged↗

Localization of annexin I (lipocortin I, p35) mRNA in normal and diseased human skin by in situ hybridization.

Annexin I is a calcium- and phospholipid-binding protein that is involved in the regulation of cellular differentiation. The aim of the present study was to determine the localization of annexin I mRNA expression in normal and diseased human skin. In situ hybridization with a specific digoxigenin-labelled RNA probe was used throughout. We detected no annexin I mRNA signals in basal and suprabasal cells of normal epidermis, but positive signals were evident in the sudoriferous ducts. Annexin I mRNA expression was detected in the keratinizing squamous cells in keratotic type seborrhoeic keratosis and in keratinocytes at the periphery of the horn pearl in well-differentiated squamous cell carcinoma. Positive signals were also seen at the border between involved and noninvolved skin in psoriasis vulgaris and in dyskeratotic epidermal keratinocytes in keratosis follicularis Darier. By contrast, no annexin I mRNA signals were detected in tumour cells in basal cell carcinoma. The present results suggest that annexin I expression is related to, and may play a role in, keratinization disorders.

Annexin A1↗

Apoptotic process of cerebellar degeneration in experimental methylmercury intoxication of rats.

This report deals with the mechanism involved in the cerebellar degeneration following experimental methylmercury poisoning of male Wistar rats. The cerebellar granule cells of animals that exhibited typical hind leg paresis were shrunken and displayed marked nuclear pyknosis. At the ultrastructural level, the nuclei of these cells were condensed and fragmented, features which are characteristic of apoptosis. In situ staining for DNA strand breaks revealed that the pyknotic nuclei were positively labeled. DNA fragmentation was confirmed by agarose gel electrophoresis; a ladder pattern of multiples of approximately 200-base pair fragments, typical of apoptosis, was observed with the cerebellar DNA of the methylmercury-treated animals. These observations suggest that the degeneration of cerebellar granule cells by alkyl mercury compounds involves an apoptotic process.

Animals↗

Modulation of erythropoietin production by selective adenosine agonists and antagonists in normal and anemic rats.

Hypoxia or anemia is the fundamental stimulus for erythropoietin (EPO) production. Recent in vitro studies suggest that EPO secretion in response to hypoxia is regulated by adenosine in the kidney. In order to examine the in vivo effect of adenosine on EPO production, we determined the effects of adenosine receptor agonists and antagonists on serum EPO concentration in normal and anemic rats. In normal rats, intravenous injection of adenosine agonists (NECA, CHA and CGS-21680) dose-dependently stimulated EPO production. Pretreatment with KW-3902, an adenosine A1 antagonist with modest A2b antagonistic action, or KF17837, an adenosine A2a antagonist, inhibited the NECA (0.1 mg/kg, i.v.)-stimulated EPO production. Anemic hypoxia, induced by 2% (v/w body weight) blood withdrawal, increased serum EPO concentration from 38 +/- 2 to 352 +/- 76 mU/ml, with the increased serum adenosine concentration in the renal vein. KF17837 (0.1 mg/kg, i.v.), but not KW-3902 (0.1 mg/kg, i.v.), inhibited the anemic hypoxia-induced increase in EPO production. The present findings support the notion that adenosine mediates the EPO production in response to hypoxia in the kidney.

Adenosine↗

DOCK180, a major CRK-binding protein, alters cell morphology upon translocation to the cell membrane.

CRK belongs to a family of adaptor proteins that consist mostly of SH2 and SH3 domains. Far Western blotting with CRK SH3 has demonstrated that it binds to 135- to 145-, 160-, and 180-kDa proteins. The 135- to 145-kDa protein is C3G, a CRK SH3-binding guanine nucleotide exchange protein. Here, we report on the molecular cloning of the 180-kDa protein, which is designated DOCK180 (180-kDa protein downstream of CRK). The isolated cDNA contains a 5,598-bp open reading frame encoding an 1,866-amino-acid protein. The deduced amino acid sequence did not reveal any significant homology to known proteins, except that an SH3 domain was identified at its amino terminus. To examine the function of DOCK180, a Ki-Ras farnesylation signal was fused to the carboxyl terminus of DOCK180, a strategy that has been employed successfully for activation of adaptor-binding proteins in vivo. Whereas wild-type DOCK180 accumulated diffusely in the cytoplasm and did not have any effect on cell morphology, farnesylated DOCK180 was localized on the cytoplasmic membrane and changed spindle 3T3 cells to flat, polygonal cells. These results suggest that DOCK180 is a new effector molecule which transduces signals from tyrosine kinases through the CRK adaptor protein.

3T3 Cells↗

Synthesis and assembly of chimeric human immunodeficiency virus gag pseudovirions.

Expression of the HIV Gag precursor in insect cells by recombinant baculoviruses results in the assembly and budding of noninfectious pseudovirions that resemble immature virus. Three strategies for packaging additional viral epitopes into pseudovirions were examined: coinfection of insect cells with individual baculoviruses encoding separate Gag and Env structural genes, inframe Gag-Env fusion proteins, and Gag-frameshift-Env fusion proteins. Electron microscopy and Western blot analysis indicated that neither the coinfection nor the inframe fusion strategies reliably produced large quantities of structurally stable chimeric pseudovirions. The frameshift fusion method utilized the retroviral Gag-Pol ribosomal frameshift mechanism for the coexpression of Gag and Gag-frameshift-Env fusion proteins. Large quantities of pseudovirions containing both the Gag and Env epitopes were produced in insect cells. Mice inoculated with the Gag-frameshift-Env pseudovirions developed cytotoxic lymphocyte responses to both HIV Gag and Env epitopes. Vaccine and immunotherapeutic applications of chimeric pseudovirions are discussed.

Animals↗

Intestinal Na+ and Cl- levels control drinking behavior in the seawater-adapted eel Anguilla japonica

To analyze drinking mechanisms in seawater teleosts, seawater-adapted eels were used as a model system. When the intestine of the eel was perfused with iso-osmotic mannitol, the eels drank sea water. However, when the perfusion medium was switched to iso-osmotic NaCl, seawater drinking was depressed. This depression was observed even after blocking NaCl absorption across the intestine by replacement of the perfusate with choline chloride or by treatment with furosemide, an inhibitor of NaCl and water absorption across the eel intestine. Furthermore, depression of drinking rate preceded an increase in urine flow by over 1 h. These results indicate that this depression is not due to a recovery of blood volume and suggest that intestinal Cl- itself inhibits drinking. Direct action of luminal Cl- on drinking behavior was further supported by the observation that perfusion with iso-osmotic NMDG-HCl, Tris-HCl, choline chloride and RbCl all inhibited seawater drinking. When NaCl in the perfusion medium was replaced with sodium acetate, sodium butyrate, sodium methylsulfate or NaSCN, the drinking rate was enhanced threefold, suggesting that Na+ itself stimulates drinking in the absence of Cl-. In the present study, concentrations of Na+ and Cl- in the swallowed fluid were also measured simultaneously. As the drinking rate was enhanced, the Na+ and Cl- concentrations in the gastrointestinal fluid were increased. On the basis of these results, it seems possible that high concentrations of Cl- in the intestine reduce the drinking rate, thus lowering esophageal Cl- concentration due to desalination of the ingested sea water. When Cl- concentration in the intestine falls below a certain level, Na+ will stimulate seawater drinking again.

Journal Article↗

Effects of KW-3902 (8-(noradamantan-3-yl)-1,3-dipropylxanthine), an adenosine A1-receptor antagonist, on urinary excretions of various electrolytes in rats.

We examined the effects of KW-3902 (8-(noradamantan-3-yl)-1,3-dipropylxanthine), an adenosine A1-receptor antagonist, on urine volume and urinary excretions of various electrolytes in saline-loaded rats, as compared with those of furosemide, trichlormethiazide (TCM), acetazolamide and amiloride. KW-3902 at doses of 0.001-1 mg/kg (p.o.) significantly increased urine volume and excretions of sodium, calcium, magnesium, chloride and bicarbonate. In addition, KW-3902 shifted urine pH to alkaline and decreased free water reabsorption. KW-3902 did not induce kaliuresis, whereas furosemide (30 mg/kg, p.o.), TCM (1 mg/kg, p.o.) and acetazolamide (25 mg/kg, p.o.) induced kaliuresis. In the KW-3902-treated group, the increases in bicarbonate excretion and urine pH were less prominent than those induced by acetazolamide, and the excretions of sodium, calcium, magnesium and chloride were similar to those induced by furosemide. The present results suggest that the adenosine A1-receptor antagonist exhibits diuresis by the inhibited reabsorption of electrolytes, not only at the proximal tubule but also at the distal tubule.

Animals↗

Purification and characterization of two lectins from calllus of Helianthus tuberosus.

Two lectins were purified from Helianthus tuberosus callus by maltose affinity chromatography and subsequent preparative electrophoresis. The lectins were designated HTA I and HTA II and their molecular masses were about 34 kDa by gel-filtration chromatography. A single band of 17 kDa and bands of 17 kDa and 18 kDa were detected after SDS-PAGE of HTA I and HTA II, respectively, indicating that HTA I is a homodimer while HTA II is a heterodimer. The amino acid compositions of the two lectins were very similar; they were rich in glycine residues, lacking detectable amounts of methionine, cysteine, and histidine. A hapten-inhibition assay showed that HTA I and HTA II had identical saccharide-binding specificity to the extent tested and belonged to the group of so-called mannose/glucose-binding lectins. They had high affinity for alpha-linked manno-oligosaccharides. Each HTA completely lost its hemagglutinating activity at pH 5.0, as a result of its dissociation to monomers, but it did not lose its ability to bind to oligosaccharides.

Amino Acids↗

Intracranial extracerebral neuroblastoma originating from the convexity dura mater.

A 19-year-old female presented with an unusual intracranial extracerebral neuroblastoma originating from the dura mater. The tumor was located in the right frontal convexity, and had adhered neither to the underlining arachnoid membrane nor the cerebral cortex. The tumor feeders were meningeal arteries. Light microscopy showed the tumor consisted of small compact cells, forming numerous Homer-Wright rosettes. Immunohistochemical examination and electron micrography confirmed the diagnosis of neuroblastoma. Intracranial extracerebral neuroblastoma is extremely rare, especially originating from the dura mater. This neuroblastoma probably originated from heterotopic neuroglial nest cells in the meninges.

Adult↗

Venous hemangioma of the temporalis muscle.

A 14-year-old girl presented with a diffuse venous hemangioma of the right temporalis muscle. The muscle had become swollen, thinning the underlying zygomatic and temporal bones. Magnetic resonance (MR) imaging demonstrated a diffuse isointense area containing high intensity foci on the T1-weighted images, and a serpiginous high intensity pattern on the T2-weighted images. A biopsy specimen revealed irregularly dilated veins in the fibrous stroma of the muscle. Diffuse abnormal signals on MR images may be a pathognomonic feature of intramuscular venous hemangiomas.

Adolescent↗

Characterization of CYP2A6 involved in 3'-hydroxylation of cotinine in human liver microsomes.

Nicotine is primarily metabolized to cotinine, and cotinine is further metabolized to trans-3'-hydroxycotinine in human liver, which is a major metabolite of nicotine in humans. We studied the formation of trans-3'-hydroxycotinine from cotinine in human liver microsomes. trans-3'-Hydroxycotinine formation demonstrated single enzyme Michaelis-Menten kinetics (Km, 234.5 +/- 26.8 MicroM; Vmax, 37.2 +/- 2.4 pmol/min/mg protein). Significant correlation (r = .967, P < .001) between cotinine 3'-hydroxylase activities at low (50 microM) and high (1 microM) cotinine concentrations in 20 human liver microsomes suggested the contribution of a single enzyme to cotinine 3'-hydroxylation. The cotinine 3'-hydroxylase activity correlated significantly with immunoreactive cytochrome P450 (CYP)2A6 contents (r = .756, P < .01) and coumarin 7-hydroxylase activity (r = .887, P < .001). The cotinine 3'-hydroxylase activity was inhibited by coumarin, alpha-naphthoflavone, chlorzoxazone and anti-rat CYP2A1 antibodies. Microsomes of B-lymphoblastoid cells expressing human CYP2A6 exhibited cotinine 3'-hydroxylase activity. The Km value of the expressed CYP2A6 (264.7 microM) was almost identical to that of human liver microsomes. In conclusion, cotinine 3'-hydroxylation appears to be catalyzed solely by CYP2A6 in humans. Cotinine is a candidate for a new substrate for CYP2A6 in humans.

Aryl Hydrocarbon Hydroxylases↗

(+)-cis-3,5-dimethyl-2-(3-pyridyl) thiazolidin-4-one hydrochloride (SM-12502) as a novel substrate for cytochrome P450 2A6 in human liver microsomes.

(+)-cis-3,5-dimethyl-2-(3-pyridyl)thiazolidin-4-one hydrochloride (SM-12502) was oxidized by human liver microsomes to produce the S-oxide as a sole metabolite. Indirect evidence suggested that the S-oxidation was catalyzed by cytochrome P450 (CYP). Eadie-Hofstee plots showed biphasic pattern, suggesting that at least two enzymes were involved in the S-oxidation in human liver microsomes. Kinetic parameters of the S-oxidase with high-affinity showed Km and Vmax values of 20.9 +/- 4.4 microM and 0.111 +/- 0.051 nmol/min/mg microsomal protein, respectively. The S-oxidase activity was inhibited by coumarin and anti-CYP2A antibody. Among the contents of forms of CYP 20 samples of human liver microsomes, the content of CYP2A6 correlated with S-oxidase activity measured with 50 microM SM-12502 (r = .808, P < .0005). A close correlation (r = .908, P < .0001) was observed between activities of SM-12502 S-oxidase and coumarin 7-hydroxylase. Microsomes from genetically engineered human B-lymphoblastoid cells expressing CYP2A6 metabolized SM-12502 to the S-oxide efficiently. The results indicate that CYP2A6 isozyme is a major form of CYP responsible for the S-oxidation of SM-12502 in human liver microsomes. Thus, SM-12502 will be a useful tool in further research to analyze a human genetic polymorphism of CYP2A6.

Animals↗

[Late coronary artery aneurysm formation following directional coronary atherectomy].

The most important factor to reduce late restenosis following directional coronary atherectomy is the technical provision of the largest luminal diameter as possible. However, the safety of deep arterial resection and the resulting arterial perforation or late aneurysm formation is questionable. This study examined the frequency of coronary artery aneurysm formation in patients treated by directional coronary atherectomy. Ninety-eight patients with 123 lesions were analyzed by quantitative angiographic and histologic data. Coronary aneurysms (ratio of dilated vessel segment to the adjacent reference segment > 1.2:1) occurred in seven patients (seven lesions: 5.7%). Specimens were categorized by the deepest layer retrieved. Forty lesions (32.5%) were treated by resection of the intima alone, 65 (52.8%) by resection of the medial tissues and 18 (14.6%) received resections of the adventitial tissues including fatty tissues in three lesions. Late coronary aneurysms occurred often in lesions with resection of adventitial tissue (38.9%) and occurred in all three lesions with resections of fatty tissues. Six coronary aneurysms occurred in the left anterior descending artery (six in segs. 6, 7), and one aneurysm in the left circumflex artery. Among the target lesions in which coronary artery aneurysms were formed, six lesions were primary lesions and one lesion was a restenotic lesion after percutaneous transluminal coronary angioplasty. Statistically, patients with coronary artery aneurysms frequently have associated hypertension (p = 0.02) but rarely diabetes mellitus (p = 0.05). Lesions with late aneurysms had significantly high specimen weight (p = 0.01) and a trend to lower postprocedure % diameter (p = 0.20). There was no significant preprocedural angiographic predictor of aneurysms such as minimal lumen diameter or reference vessel diameter.

Adult↗

[Left cardiac output curve and pulmonary venous return curve in patients with various heart diseases].

This study assessed the cardiac function of humans by drawing simultaneous left cardiac output and pulmonary venous return curves using radionuclide angiocardiography and right heart catheterization which allows recording of the pressure-flow relationship. Thirty-one subjects with various cardiac diseases were divided into two groups [18 patients with New York Heart Association (NYHA) functional class I and 13 patients with NYHA classes II or III]. Mean pulmonary filling pressure (Pmp) was calculated from the formula of Guyton, using pulmonary arterial compliance which was measured by Reuben's method and pulmonary venous compliance measured as reported previously. On the pressure-flow plane, the down slope of the pulmonary venous return curve was drawn by joining the points of (Pmp, 0) and (mean pulmonary capillary wedge pressure, cardiac output). To construct the cardiac output curve, two levels of lower body negative pressure were used to regulate the venous return to the heart. Pmp and the resistance to pulmonary venous return in NYHA II or III patients were significantly higher than those in NYHA I patients (Pmp: 16.3 +/- 1.5 vs 9.0 +/- 0.5 mmHg, p < 0.01; resistance to pulmonary venous return: 0.75 +/- 0.09 vs 0.43 +/- 0.04 mmHg/l/min, p < 0.01, respectively). The slope of pulmonary venous return curve in NYHA II or III patients was smaller than that in NYHA I patients and the pulmonary venous return curve in NYHA II or III patients shifted rightward. The slope of cardiac output curve in NYHA II or III patients was significantly smaller than that in NYHA I patients. This curve in NYHA II or III patients shifted downward and rightward. These results indicate that simultaneous cardiac output and pulmonary venous return curves may be a useful method for assessing the cardiac function in patients with various heart diseases.

Adult↗

Analysis of the hepatocyte growth factor receptor in regeneration and oncogenesis of the liver.

The c-met proto-oncogene product is the tyrosine kinase receptor for hepatocyte growth factor. To gain insights into its functions in the liver, we carried out a study of the expression and tyrosine phosphorylation status of the Met protein during hepatic regeneration and in human hepatocellular carcinomas. Following partial hepatectomy of rats, decreased expression of the proreceptor p170MET and reduced levels of tyrosine phosphorylation were seen within 12 h. These changes were still evident 7 days later. By contrast, there was no significant alteration of the Met beta subunit. The analysis of samples from 18 patients with hepatocellular carcinoma revealed that the expression of p160MET proreceptor was increased in non-cancerous areas, while that of the p145 beta MET proreceptor was significantly greater in the tumor tissue than in the non-tumor areas (p < 0.05). These findings suggest that processing of the Met proreceptor is closely associated with regeneration and carcinogenesis of the liver.

Adult↗

[A case of spinal epidermoid caused by lumbar punctures].

A case of spinal epidermoid caused by lumbar punctures is reported. The 50-year-old man, who had been diagnosed as having poliomyelitis and who had been treated with lumbar puncture 24 times when he was 1 year old, came to our clinic complaining of gradual worsening of atrophy and weakness in his right leg during the last 10 years. Neuroradiologically, intraspinal tumor with calcification was detected in the lumbar region and partial removal of the tumor was performed. Pathological diagnosis was that it was epidermoid. During frequent lumbar puncture at the age of 1, some small skin grafts had been implanted intrathecally and we concluded that, changing its original nature, it had grown slowly to become epidermoid. We want to emphasize especially here that, when we performe lumbar puncture, we should, as far as possible, use a small needle with stilette so as not to induce future spinal epidermoid.

Epidermal Cyst↗

Role of human cytochrome P4502A6 in C-oxidation of nicotine.

Nicotine is primarily metabolized to cotinine in humans. In this study, human cytochrome P450 (CYP) isoform involved in cotinine formation was identified. The formation of cotinine in 16 human liver microsomes was determined with a 50 microM nicotine concentration and with a cytosol preparation as a source of aldehyde oxidase. Cotinine formation in human liver microsomes significantly correlated with immunochemically determined CYP2A6 levels (r = 0.663, p < 0.05), coumarin 7-hydroxylase activities (r = 0.831, p < 0.01), and cotinine 3'-hydroxylase activities (r = 0.735, p < 0.01) that are responsible for CYP2A6. In inhibition studies, cotinine formation in human liver microsomes was inhibited by coumarin and rabbit anti-rat CYP2A1 antibody specifically. When the capability of microsomes of B-lymphoblastoid cells expressing human CYPs to perform biotransformation of nicotine to cotinine was determined, cDNA-expressed CYP2A6 exhibited the highest cotinine formation. The KMapp values from microsome expressing CYP2A6 cDNA were similar to the value obtained from human liver microsomes. The large interindividual variabilities in cotinine formation and immunochemically determined CYP2A6 levels were observed in human liver microsomes, suggesting genetic polymorphism of CYP2A6. Nicotine is a new in vivo probe for phenotyping of CYP2A6 in humans.

Aryl Hydrocarbon Hydroxylases↗