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Biomedical subjects

H Nagae

Publications and source records attributed to H Nagae.

At least 19 recordsLinked to original sources
PubMed1995

Preparation and characterization of dextran magnetite-incorporated thermosensitive liposomes: an on-line flow system for quantifying magnetic responsiveness.

PURPOSE: Dextran magnetite (DM)-incorporated thermosensitive liposomes, namely thermosensitive magnetoliposomes (TMs), were prepared and characterized in order to investigate their possibility for magnetic drug targeting. METHODS: TMs containing calcein were prepared at various DM concentrations by reverse-phase evaporation of dipalmitoylphosphatidylcholine (DPPC). They were evaluated for their physicochemical properties including size, DM capture, magnetite distribution within liposomes, and temperature-dependent calcein release. Moreover, a novel on-line flow apparatus with a sample injector, a coil of tubing placed in an electromagnet, and a fluorescence detector was developed for quantifying the magnetic responsiveness of TMs. This device allowed us a real-time measurement of percentage holding of TMs by magnetic field. RESULTS: Due to water-soluble property of DM, higher contents of magnetite up to 490 mg per mmol DPPC were successfully incorporated into the liposomes with DM than with conventional magnetite (Fe3O4). Thermosensitivity and lipid integrity of TMs were not influenced by inclusion of DM. Using the on-line flow system, percentage holding of TMs by magnetic field was shown to vary with several factors; it increases as the magnetic field strength increases, the fluid flow rate decreases, the magnetite content increases, and the liposome concentration increases. Typically, at 490 mg incorporated magnetite per mmol DPPC, 0.5 ml/min-fluid flow rate, and high magnetic field strength (> or = 10 kiloGauss), approximately 100% of TMs were found to be held. CONCLUSIONS: The TMs were suggested to be useful in future cancer treatment by magnetic targeting combined with drug release in response to hyperthermia.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1995

Serial cultivation of human nail matrix cells under serum-free conditions.

We have established serial cultures of human nail matrix cells (NMCs) under serum-free conditions. We cultured NMCs using two different methods depending upon the volume of nail matrix obtained. When a sufficient amount of nail matrix was obtained, they were minced and treated with 0.25% trypsin and 0.03% EDTA. The NMCs were transferred directly as a dispersed cell culture into KGM medium. Because a sufficient amount of matrix was rarely obtained, we developed a method by which NMCs were cultured primarily as implanted small matrices in Eagle's MEM (high Ca+ medium) supplemented with 15% fetal bovine serum for the first 4 to 5 days; during this time, the NMCs expanded from the matrices and formed colonies around them. NMCs then were cultured with KGM. In both methods, KGM medium supported the growth of NMCs without a biological feeder layer. These cells could be cultivated serially for at least seven passages. Half of the cells were positively stained with a monoclonal antibody against hair (hard) keratin which is expressed in nail matrix in vivo, indicating that the cells originated from the nail matrix. These methods will now permit investigations of nail matrix cells that previously were unfeasible because of the relative lack of cells and difficulties with propagation.

Animals↗
PubMed1995

Maternal plasma endothelin levels and fetal status in normal and preeclamptic pregnancies.

Endothelin (ET) is a potent vasoconstrictor peptide. In this study, we investigated maternal venous plasma ET levels measured by Sandwich-enzyme immunoassay within a week before the onset of labor, and measured plasma renin activity and plasma aldosterone concentration by radioimmunoassay in normal and severely preeclamptic pregnancies. Also, we determined umbilical cord blood pH and gas concentrations after spontaneous vaginal deliveries and cesarean sections. There was a significant (p < 0.01) negative correlation between maternal ET levels within 1 week before the onset of labor and birth weights. There was no significant correlation between maternal ET levels and umbilical gas concentrations. These data suggest that the correlation is the result of decreasing uteroplacental blood flow. We speculate that increased maternal ET expresses not only maternal renal vascular endothelial injury but also other vascular endothelial injuries. These vascular injuries may occur at least 1 week before the clinical manifestation in the preeclamptic mothers and their fetuses.

Adult↗
PubMed1995

Pulmonary surfactant protein D in sera and bronchoalveolar lavage fluids.

Pulmonary surfactant protein D (SP-D) is a hydrophilic glycoprotein with a reduced molecular mass of 43 kDa and a member of the C-type lectin superfamily, along with mannose-binding proteins and surfactant protein A (SP-A). We have recently prepared monoclonal antibodies against human SP-D and developed an enzyme-linked immunosorbent assay (ELISA). In this study, the levels of SP-D in sera and bronchoalveolar lavage (BAL) fluids of patients with lung diseases were determined by ELISA, using human recombinant SP-D as a standard. We demonstrated that the concentrations of SP-D in sera are prominently increased in patients with idiopathic pulmonary fibrosis (IPF), interstitial pneumonia with collagen disease (IPCD), and pulmonary alveolar proteinosis (PAP). Patients with IPF, IPCD, and PAP exhibited levels of serum SP-D 5.1-fold, 7.2-fold, and 7.0-fold, respectively, of those in healthy volunteers; 91.5% of the patients with IPF, 81.3% with IPCD, and 100% with PAP exhibited serum SP-D levels that exceeded the cut-off value (mean + 2 SD of control value). Serum SP-D levels appeared to reflect the disease activity of IPF and IPCD and the disease severity of PAP. High levels of SP-D in BAL fluids were shown in patients with PAP, but not with IPF and IPCD. We conclude that measurement of SP-D in sera can provide an easily identifiable and useful clinical marker for the diagnosis of IPF, IPCD, and PAP, and can predict the disease activity of IPF and IPCD and the disease severity of PAP.

Adult↗
PubMed1994

Brain natriuretic peptide and atrial natriuretic peptide levels in normal pregnancy and preeclampsia.

Brain natriuretic peptide (BNP) was increased in many hypertensive subjects. In this study, we have evaluated maternal, umbilical plasma and amniotic fluid BNP and atrial natriuretic peptide (ANP) in 19 normotensive pregnant women and in 35 preeclamptic patients. The maternal plasma and umbilical cord plasma ANP (p < 0.05) and BNP (p < 0.005) levels were significantly higher than those in normal pregnancy. There was no significant correlation among ANP level, BNP level, clinical symptoms and laboratory examinations. It is suggested that ANP and BNP may be rather a sequel to preeclamptic pathophysiological changes, and may not play an important role as the etiological factor of preeclampsia.

Amniotic Fluid↗
PubMed1994

Serum collagen IV and laminin levels in preeclampsia.

Collagen IV is the main collagenous component localized in the trophoblast and glomerular basement membrane. Serum collagen V reflects degradation of basement membrane collagen. In this study, we measured collagen IV levels in maternal serum, umbilical cord serum and amniotic fluid, both from preeclamptic and normal pregnant women, by radioimmunoassay. The serum collagen IV levels in the preeclamptic group were significantly (p < 0.05) higher than those in the normal pregnant group. The amniotic fluid collagen IV level at term was found to be higher than maternal serum collagen IV. We postulate that collagen IV may have an important role in the maintenance of pregnancy. There was a significant positive correlation between maternal serum collagen IV levels and serum laminin levels. There was no significant correlation between maternal serum collagen IV level and blood pressure, urinary protein concentration, or any other laboratory data. These results suggest that there is early damage of endothelial cells in preeclampsia.

Amniotic Fluid↗
PubMed1993

Expression of anticardiolipin cofactor, human beta 2-glycoprotein I, by a recombinant baculovirus/insect cell system.

A full-length cDNA coding a human beta 2-glycoprotein I (beta 2-GPI) was introduced into the baculovirus genome to construct a recombinant baculovirus. Spodoptera frugiperda (Sf9) cells were infected with the recombinant baculovirus. A protein (mol. wt 43,000) reactive with anti-beta 2-GPI antisera was produced in the insect cells and secreted into the culture medium. The recombinant beta 2-GPI was purified from the culture supernatant by sequential cardiolipin (CL)-affinity column chromatography and gel filtration. The N-terminal amino acid sequence of the protein was identical to that of the native beta 2-GPI purified from human sera, and a putative signal peptide was cleaved from the secreted form of the recombinant protein. The purified recombinant protein had a cofactor activity which enhances CL binding of anticardiolipin antibodies (aCL) in systemic lupus erythematosus (SLE) patients, as well as the native beta 2-GPI. Thus, the beta 2-GPI expressed in insect cells is an immunologically active cofactor.

Amino Acid Sequence↗
PubMed1993

Protection by L-ascorbic acid against phototoxicity in tin-protoporphyrin-treated suckling rats.

The protective effect of free radical scavengers against phototoxicity was investigated with tin-protoporphyrin (SnPP)-treated suckling rats. Six kinds of scavengers (L-ascorbic acid, reduced glutathione, alpha-tocopherol, retinol, uric acid and cystine) were intraperitoneally injected to rats treated with SnPP plus photoirradiation. Among them, L-ascorbic acid was found to be most effective in protecting SnPP-treated rats against phototoxicity. The survival period was markedly prolonged, and the frequency of abnormal behaviors was reduced with the treatment. Lipid peroxidation in vitro with the brain membrane fraction was also suppressed. The other five substances gave only a little antioxidant effect both in vivo and in vitro. The present study shows that L-ascorbic acid may be a promising chemical to prevent the phototoxicity of SnPP.

Animals↗
PubMed1993

Serum laminin levels in normal pregnancy and preeclampsia.

Laminin is a large noncollagenous glycoprotein localized in the trophoblast and glomerular basement membrane. We measured laminin levels in maternal serum, umbilical cord serum and amniotic fluid, both from preeclamptic and normal pregnant women, by enzyme immunoassay. The serum laminin levels in the preeclamptic group were significantly (p < 0.05 to p < 0.01) higher than those in the normal pregnant group. It has been suggested that laminin plays an important role in implantation of the placenta during early pregnancy. In this study, the amniotic-fluid laminin level at term was found to be lower than maternal serum laminin. We postulate that laminin may not have an important role in the maintenance of late pregnancy. There was a significant positive correlation between maternal serum laminin levels and serum uric acid levels. There was no significant correlation between maternal serum laminin level and blood pressure, urinary protein concentration, or any other laboratory data. These results suggest that there is damage of glomerular and placental spiral arteries in preeclampsia.

Female↗
PubMed1993

Effects of thromboxane synthetase inhibitor (OKY-046) on placental blood flow, placental weight and fetal weight in normotensive and spontaneously hypertensive rats.

Recently, thromboxane synthetase inhibitor has been used for the treatment of preeclampsia. In this study, we investigated the effects of thromboxane synthetase inhibitor (OKY-046) on placental blood flow (measured with clearance of hydrogen gas generated by electrolysis) in normotensive Wistar-Kyoto rats (WKY) and spontaneously hypertensive rats (SHR). The systolic blood pressure of OKY-046-treated SHR (1, 4 and 8 mg/kg) decreased significantly (p < 0.05); however, the systolic blood pressure of WKY did not decrease. The placental blood flow of both OKY-046-treated WKY and SHR did not decrease. We found that OKY-046 has no reducing effect on placental blood flow in rats, and systolic blood pressure of SHR decreases. These data suggest that thromboxane synthetase inhibitor might have a beneficial effect on preeclampsia.

Animals↗
PubMed1992

Immunohistological analysis of P53 expression in human skin tumors.

The p53 expression in various skin tumors was immunohistologically evaluated using two mouse monoclonal anti-p53 antibodies, PAb421 and PAb1801. The p53 expression was not detected in the normal epidermal cells. Nuclear staining suggested that the p53 expression was observed in 10 of 26 squamous cell carcinomas (SCCs) from 24 patients, in one undifferentiated carcinoma, one proliferating trichilemmal cyst, one malignant proliferating trichilemmal tumor and in one metastatic carcinoma of breast cancer. None off four cases of Bowen's disease (SCC in situ) showed nuclear staining. In the SCCs, five of 20 primary lesions, three of four recurrent lesions and both of two metastatic lesions had positive nuclei. There was one case of SCC in which a primary lesion was negative but a recurrent lesion was positive. Thus, p53 expression was more frequently observed in SCCs at more clinically advanced stages. This may suggest that p53 has some relevance to progression of SCC. Nuclear staining was not detected in any of the following cases: two cases of seborrheic keratosis, one eccrine poroma, one keratoacanthoma, 11 basal cell epitheliomas, two mammary Paget's disease, three genital Paget's disease, one sebaceous carcinoma, four malignant melanomas, six lymphomas, two leukemia cutis and two angiosarcomas.

Antibodies, Monoclonal↗
PubMed1992

Effects of nifedipine on placental blood flow, placental weight and fetal weight in normotensive and spontaneously hypertensive rats.

The effect of nifedipine on placental blood flow was investigated in spontaneously hypertensive rats (SHR) and compared with that in normotensive Wistar Kyoto rats (WKY) by means of the clearance of hydrogen gas generated by electrolysis. The placental blood flow of nifedipine-treated WKY (5, 10 and 25 mg/kg) was significantly (p < 0.05) reduced compared with that of WKY control. On the other hand, the placental blood flow of nifedipine-treated SHR did not change compared with that of SHR control. These data suggest that nifedipine has different effects on placental blood flow in SHR and WKY.

Animals↗
PubMed1992

Purification and some properties of a Haim-sensitive alpha-amylase from newly isolated Bacillus sp. No. 195.

Newly isolated Bacillus sp. No. 195 produced an extracellular alpha-amylase sensitive to Haim which was found to inhibit specifically animal alpha-amylases. The enzyme was purified easily by two steps of starch adsorption and gel filtration using Sephacryl S-200. The purified enzyme, which showed a single band on native-PAGE or SDS-PAGE, had a molecular weight of 60,000 as judged on SDS-PAGE. The optimum pH value for activity and the isoelectric point were around 7.0 and 4.5, respectively. The sensitivity of the amylase to Haim was similar to that of animal amylase rather than bacterial amylase. It was suggested that a Haim-amylase complex might be formed at the molar ratio of 1:1. The amino acid sequence F-S-W similar to the triplet F-E-W highly conserved among alpha-amylases sensitive to proteinaceous inhibitors, such as Hoe 467-A or Haim, was found in the amino-terminal part of the No. 195 amylase.

Amino Acid Sequence↗
PubMed1992

[Hyperthermia for cancer with dextran magnetite using tubular implants].

Dextran magnetite particles (Meito Sangyo Corp.) are an aqueous magnetite zol and a nanometer complex consisting of dextran chains surrounding a core of ultrafine iron oxide. The tubular implants were made with polyester tube (3 mm diameter, 20 mm length) filled with DM aqueous zol (29%w/v). The temperature of an agar phantom with implants was measured in the inductive field. Heating was effected by creating an electromagnetic field with a 7 kW generator operating at 500kHz (Yamamoto Vinyter). The temperature was elevated 3.4 degrees at a distance of 5 mm from the implant at an inductive power of 2.6 kW. An area of 20 x 20 mm was heated while changing the power, the number of implants, and their arrangement. Selective heating of cancer was considered possible by inductive heating at 500 kHz and DM implants. Since the DM aqueous zol configuration can be readily changed, treatment of various cancers is possible.

Dextrans↗
PubMed1991

Molecular definition of human beta 2-glycoprotein I (beta 2-GPI) by cDNA cloning and inter-species differences of beta 2-GPI in alternation of anticardiolipin binding.

Human beta 2-glycoprotein I (beta 2-GPI) is involved in cardiolipin (CL) binding of anticardiolipin antibodies (aCL) in systemic lupus erythematosus (SLE). We examined the inter-species differences of beta 2-GPI in alternation of CL binding of aCL. beta 2-GPI preparations were obtained from human, bovine, and rat sera by sequential CL--polyacrylamide affinity, DEAE--cellulose, and anti-human IgG-conjugated Sepharose CL-4B column chromatography, and they had apparent molecular weights of 50, 53, and 55 kDa respectively. Human beta 2-GPI not only enhanced CL binding by aCL in SLE but also depressed it by those in syphilis. Either bovine and rat beta 2-GPI exerted no or quite small inhibition of the binding of syphilitic aCL compared with human beta 2-GPI whereas all three species of beta 2-GPI generated binding of aCL in SLE to a similar degree. Further, a complete cDNA clone, p beta 2-GPI, was isolated from a human hepatoma cell line, HepG2, and its nucleotide sequence was analyzed. The sequences of bovine and rat counterpart molecules (beta 2-GPI) are highly homologous to that of the deduced sequence, and their corresponding regions are 84.0 and 82.5% identical to the complete domain and to the amino acid sequence 53-326 of human beta 2-GPI respectively. One of major differences appears at position 154 in human beta 2-GPI, and might be associated with the inhibitory effect on the binding of syphilitic aCL. The sequencing analysis of these beta 2-GPI proteins might provide leads to functional sites of domains which would be associated with such serological phenomena.

Amino Acid Sequence↗
PubMed1991

Sn-protoporphyrin plus photoirradiation induces lipid peroxidation in vivo and in vitro in nonjaundiced Gunn rats.

Lipid peroxidation induced by Sn-protoporphyrin (SnPP) plus photoirradiation was investigated in vivo and in vitro using nonjaundiced Gunn rats. Membrane lipids from young adult rat brain were peroxidized by SnPP plus photoirradiation depending on the SnPP concentration and photoirradiance. Similarly, coadministration of SnPP and photoirradiation to suckling rats increased lipid peroxides in the whole blood and was found lethal. The influence of the wavelength distribution of light sources was also examined by using blue-white and green fluorescent lights. The photodynamic effect by green light irradiation whose energy distribution had no overlap with the Soret band of SnPP was about half of that produced by blue-white light with regard to the membrane peroxidation and the lethal effect on neonatal rats. We therefore conclude that the combination of SnPP and photoirradiation is a potentially hazardous treatment of neonatal jaundice.

Animals↗
PubMed1991

Maternal and fetal atrial natriuretic peptide levels, maternal plasma renin activity, angiotensin II, prostacyclin and thromboxane A2 levels in normal and preeclamptic pregnancies.

To clarify the possible role of elevated atrial natriuretic peptide (ANP) in the pathophysiology of preeclampsia, we measured ANP, renin activity (PRA), angiotensin II (Ang II), TXB2 (a stable metabolite of TXA2) and 6-keto-PGF1 alpha (a stable end product of PGI2) concentrations in the plasma of 19 normal pregnant women and 35 severe preeclamptic patients at term. Plasma ANP levels in the preeclamptic patients (n = 35, 71.5 +/- 3.8 pg/ml, mean +/- S.E.) and also umbilical plasma ANP (n = 35, 83.0 +/- 4.2 pg/ml) were significantly (p less than 0.01) higher than those of normal pregnant women plasma (n = 19, 58.7 +/- 3.7 pg/ml) and umbilical plasma (n = 19, 47.6 +/- 4.7 pg/ml). There was a significant (p less than 0.01) positive correlation between maternal ANP levels and fetal ANP levels (n = 54, r = 0.44). Plasma PRA and 6-keto-PGF1 alpha levels in preeclampsia were significantly (p less than 0.05) lower than those of normal pregnancy. The ratio of 6-keto-PGF1 alpha/TXB2 in preeclampsia was significantly (p less than 0.01) lower than that of normal pregnancy as we reported previously. There was no significant correlation between plasma ANP level and plasma PRA, Ang II, plasma TXB2 and 6-keto-PGF1 alpha concentrations. Moreover there was no significant correlation between plasma ANP level and the severity of preeclampsia. These data suggest the possibility of a transplacental crossing of ANP secreted by feto-placental unit, which might be, at least in part, responsible for the high ANP levels observed in preeclampsia. The ANP in preeclampsia is not related directly to hypertension, but it may play a substantial role in the regulation or normalization of blood volume and vascular reactivity.

6-Ketoprostaglandin F1 alpha↗