Biomedical subjects

S Goya

Publications and source records attributed to S Goya.

At least 19 recordsLinked to original sources

PubMed2001

Microsatellite instability in transforming growth factor-beta 1 type II receptor gene in alveolar lining epithelial cells of idiopathic pulmonary fibrosis.

It has been reported that transforming growth factor (TGF)-beta, which plays an integral role in the pathogenesis of idiopathic pulmonary fibrosis (IPF), suppresses proliferation of alveolar epithelial cells in vitro. Although hyperplastic lesions of alveolar lining epithelial cells (ALECs) are characteristic pathologic features of IPF, the mechanism of their involvement in the pathogenesis has not yet been extensively studied. On the assumption that the hyperplastic ALECs have escaped from the growth-inhibitory effects of TGF-beta, we searched for mutations in the microsatellite of the TGF-beta receptor type II (T beta RII) gene. To detect a deletion in the polyadenine tract in exon 3 of the T beta RII gene, cells were isolated by microdissection from lung sections of IPF patients, and DNA was extracted from these cells and amplified by high-fidelity polymerase chain reaction. A total of 121 sites of hyperplastic ALECs from 11 IPF patients were analyzed, and a one-base-pair deletion was detected in nine sites from five patients. The mutation was also detected in smooth muscle-like cells of the thickened pulmonary artery. In some tissue areas where the deletion was detected, low T beta RII expression was confirmed by immunohistochemical staining. These data suggest that microsatellite instability in the T beta RII gene occurred in some lesions of hyperplastic ALECs in IPF, although at a low incidence, and that this genetic disorder might play a partial role in the pathologic changes of IPF.

Adult↗

PubMed2000

Introduction of the interleukin-10 gene into mice inhibited bleomycin-induced lung injury in vivo.

Interleukin (IL)-10 has been shown to reduce many inflammatory reactions. We investigated the in vivo effects of IL-10 on a bleomycin-induced lung injury model. Hemagglutinating virus of Japan (HVJ)-liposomes containing a human IL-10 expression vector (hIL10-HVJ) or a balanced salt solution as a control (Cont-HVJ) was intraperitoneally injected into mice on day -3. This was followed by intratracheal instillation of bleomycin (0.8 mg/kg) on day 0. Myeloperoxidase activity of bronchoalveolar lavage fluid and tumor necrosis factor-alpha mRNA expression in bronchoalveolar lavage fluid cells on day 7 and hydroxyproline content of the whole lung on day 21 were inhibited significantly by hIL10-HVJ treatment. However, Cont-HVJ treatment could not suppress any of these parameters. We also examined the in vitro effects of IL-10 on the human lung fibroblast cell line WI-38. IL-10 significantly reduced constitutive and transforming growth factor-beta-stimulated type I collagen mRNA expression. However, IL-10 did not affect the proliferation of WI-38 cells induced by platelet-derived growth factor. These data suggested that exogenous IL-10 may be useful in the treatment of pulmonary fibrosis.

Animals↗

PubMed1999

In vivo gene transfer of an extracellular domain of platelet-derived growth factor beta receptor by the HVJ-liposome method ameliorates bleomycin-induced pulmonary fibrosis.

A number of investigators have reported augmented expression of PDGF in lungs with idiopathic pulmonary fibrosis (IPF) or with other types of pulmonary fibrosis. To accomplish such a regulation of PDGF activity, we constructed an expression plasmid of the extracellular domain of PDGF receptor beta chain (XR), which lacks intracellular tyrosine kinase domain and transmembrane portions, and estimated the therapeutic effects of XR gene transfer through the trachea on bleomycin-induced lung fibrosis of C57BL/6 mice using the hemagglutinating virus of Japan(HVJ)-liposome method. The XR gene transfer ameliorated the increases in the wet weight and hydroxyproline content and the histopathologic changes of the lung induced by bleomycin. These findings suggest that PDGF plays a crucial role in the pathogenesis of pulmonary fibrosis, and that XR gene transfer using the HVJ-liposome method may limit the progression of pulmonary fibrosis.

Animals↗

PubMed1999

Effects of in vivo soluble selectin gene introduction of lps-induced leukocyte accumulation in the murine lung.

The selectin family adhesion molecules exert a crucial role in accumulation of leukocytes at the site of inflammation. To test the biological effects of soluble selectin on lung inflammation, we introduced an expression plasmid vector of soluble selectin gene via HVJ-liposome into a murine model of LPS-induced lung injury. The myeloperoxidase activity in LPS-injected mice was suppressed by the in vivo injection of soluble P-selectin gene relative to control mice. On the contrary, soluble E- or L-selectin genes did not exert suppressive effects. Our observations suggest that P-selectin plays a crucial role in the initial steps of lung inflammation, and exogenous introduction of soluble P-selectin by in vivo gene transfer method may be a useful strategy for regulating inflammation of the lung.

Animals↗

PubMed1999

Introduction of plasmid DNA and oligonucleotides into lung epithelial cells by the hemagglutinating virus of Japan (HVJ)-liposome method.

The hemagglutinating virus of Japan (HVJ) fused with liposomes provides a unique transfection vehicle with characteristics of both virus vector and liposome. Here we investigate the efficiency and safety of the HVJ-liposome technique in delivering foreign genes and oligonucleotides into the lung of the Wistar rat. A plasmid vector containing the Escherichia coli beta-galactosidase (beta-gal) gene and the chicken beta-actin promoter was transfected via the trachea using the HVJ-liposome method. Cytochemical staining showed expression of exogenous beta-gal activity in airway epithelial cells, alveolar macrophages, and alveolar type II cells. This activity persisted at least 28 days after administration of the genes. FITC-labeled oligonucleotides also were introduced into the same types of lung cells as those expressing beta-gal. After instillation of HVJ-liposome, anti-HVJ antibodies were detected in the sera of the rats, but even after repeated administration of HVJ-liposome, no marked histopathologic change was observed while exogenous beta-gal expression was detected in pulmonary cells.

Actins↗

PubMed1996

Methylphenidate and attentional training. Comparative effects on behavior and neurocognitive performance in twin girls with attention-deficit/hyperactivity disorder.

The effectiveness of four doses (5-mg, 10-mg, 15-mg, 20-mg) of methylphenidate (MPH) and attentional training (AT) were evaluated using neurocognitive instruments (Continuous Performance Test; Matching Unfamiliar Figures Test), narrow- and broad-band rating scales in the context of a double-blind, placebo-control, within-subject reversal design for dizygotic twin girls with Attention-Deficit/Hyperactivity Disorder (ADHD). Both interventions proved effective for improving neurocognitive test performance and behavior, although broad-band ratings revealed dose-response curves different from those obtained from the neurocognitive tests. Implications for clinical management of girls with ADHD are discussed.

Attention↗

PubMed1993

[Determination of amantadine by pre-labeling with 3-(7'-methoxycoumarin-3'-carbonyl)-benzoxazoline-2-thione and high-performance liquid chromatography with fluorescence detection].

3-(7'-Methoxycoumarin-3'-carbonyl)-benzoxazoline-2-thione (MCBT) was synthesized as a precolumn fluorescent labeling reagent for amantadine for use in high-performance liquid chromatography (HPLC). Amantadine was derivatized quantitatively into a fluorescent compound through the amino group by treatment with MCBT at room temperature for 5 min in aqueous 95% acetonitrile solution in the presence of 4-dimethylaminopyridine. The derivative was subjected to HPLC on TSK gel ODS-80TM (250 x 4.6 mm i.d.) with methanol-water (10:1) as the mobile phase and monitored with an excitation wavelength of 355 nm and an emission wavelength of 405 nm. The limit of detection was 2.0 micrograms/ml in the urine. This method was satisfactory with respect to simplicity and precision to quantify amantadine spiked in the urine.

Amantadine↗

PubMed1991

7-Alkylaminocoumarin-4-acetic acids as fluorescent probe for studies of drug-binding sites on human serum albumin.

7-Alkylaminocoumarin-4-acetic acids I-IX having alkylamino groups different in alkylchain lengths were synthesized as fluorescence probes for characterization of drug-binding sites on human serum albumin (HSA). The fluorescences of I-IX were quenched or enhanced in the presence of HSA with shifts of the emission maxima to shorter wavelength. The binding constants and the number of binding sites were determined by the spectral changes of the probes I-IX bound to HSA through analysis of Scatchard's and Job's plots. The primary binding sites of the tested probes were found to be site 2 (diazepam site) on HSA from the results of competitive displacement studies. The polarity of site 2 was estimated from the relationship between the emission maximum of the probe of IV and Z-values, and was found to be comparable to that of acetonitrile. Simple attempts to estimate the site 2 region from the molecular size of the probe of VIII obtained using the Corey-Pauling-Koltun molecular model suggest that the hydrophobic cleft at site 2 is about 21-25 A in depth. The distance between the lone tryptophan residue in HSA and probes bound to site 2 was estimated to be 15-17 A using Förster's equation on the basis of fluorescence energy transfer. The present data suggest that I-IX are useful as fluorescence probes for the characterization of site 2 on HSA.

Acetates↗

PubMed1991

[Fluorescence label of alcohols and amines with pyrene-1-carbonyl fluoride].

Pyrene-1-carbonyl fluoride (PCF) was synthesized as a precolumn fluorescent labeling reagent for alcohols and amines for use in high performance liquid chromatography (HPLC). PCF reacted with primary and phenolic hydroxyl groups in dichloromethane at 100 degrees C for 30 min in the presence of 4-dimethylaminopyridine (DMAP) to give the corresponding fluorescent pyrene esters and also reacted with primary amines in acetonitrile at room temperature for 2 min in the presence of DMAP to give the amides. The PCF esters of corticosteroids were separated by normal-phase chromatography on a Cosmosil 5 SL column with hexane-ethyl acetate (6:5, v/v) and the PCF amides of primary amines were separated by reversed-phase chromatography on a TSK gel ODS-80 TM column with methanol-water (10:3, v/v). The detection limits (S/N = 3) of cortisone and 2-phenylethylamine were 600 fmol and 800 fmol for an injection volume of 10 microliters, respectively.

Alcohols↗

PubMed1989

[Synthesis of 1-(difluoro-1,3,5-triazinyl)-2-methylisoindole as a fluorescent derivatization reagent and its reactivity for corticosteroids].

A fluorescence high-performance liquid chromatographic method for the determination of corticosteroids is described. Corticosteroids were labeled quantitatively by treatment with 1-(difluoro-1,3,5-triazinyl)-2-methylisoindole in an alkaline medium at room temperature for 10 s and the fluorescent adducts were eluted isocratically on a reversed-phase column by a mobile phase of aqueous methanol. The detection limit of cortisone was 3 pmol for an injection volume of 10 microliter.

Chromatography, High Pressure Liquid↗

PubMed1989

[Synthesis and reactivity of anthracene-1-carbonyl azide as a fluorescent derivatization reagent for alcohols].

Anthracene-1-carbonyl azide was found to be a sensitive fluorescent derivatization reagent for primary and secondary alcohols for high-performance liquid chromatography. Reaction conditions were investigated for cholestanol. The reagent reacts with alcohols in benzene in the presence of 4-dimethylaminopyridine to give the corresponding fluorescent urethanes at 100 degrees C for 50 min. The detection limit for cholestanol was 250 fmol for an injection volume of 10 microliters. Tertiary alcohols and phenols do not produce fluorescent derivatives under these conditions.

Alcohols↗

PubMed1988