Biomedical subjects

S Nagai

Publications and source records attributed to S Nagai.

At least 91 records · Page 5Linked to original sources

PubMed1999

Smoke extract stimulates lung fibroblasts to release neutrophil and monocyte chemotactic activities.

Accumulation of monocytes and neutrophils and fibrous distortion of the airway are characteristics of airway disease secondary to smoking. The presence of inflammatory cells and fibrosis correlate, and, therefore, we postulated that lung fibroblasts might release chemotactic activity for neutrophils and monocytes in response to smoke extract. To test this hypothesis, human fetal lung (HFL1) fibroblasts were cultured, and the supernatant fluid was evaluated for neutrophil (NCA) and monocyte (MCA) chemotactic activities with a blind well chamber technique. HFL1 fibroblasts released chemotactic activity in response to smoke extract in a dose- and time-dependent manner (P < 0.05). Checkerboard analysis showed that the activity was predominantly chemotactic. Partial characterization of the released chemotactic activity revealed that the activity was partly heat labile, trypsin sensitive, and ethyl acetate extractable. Lipoxygenase inhibitors and cycloheximide inhibited the release of both NCA and MCA. Molecular-sieve chromatography revealed that NCA and MCA were heterogeneous. NCA was inhibited by anti-human interleukin (IL)-8 and anti-granulocyte colony-stimulating factor antibodies and a leukotriene (LT) B(4)-receptor antagonist. Anti-granulocyte-macrophage colony-stimulating factor (GM-CSF) and anti-monocyte chemoattractant protein (MCP)-1 antibodies and an LTB(4)-receptor antagonist inhibited MCA. Immunoreactive IL-8, granulocyte colony-stimulating factor, GM-CSF, and MCP-1 significantly increased in culture supernatant fluid in response to smoke extract. Finally, smoke extract augmented the expression of mRNAs of IL-8, GM-CSF, and MCP-1. These data demonstrate that lung fibroblasts release NCA and MCA in response to smoke extract and suggest that lung fibroblasts may modulate the inflammatory cell recruitment into the lung.

Cells, Cultured↗

PubMed1999

The number of interleukin 1 receptors on lung fibroblasts in patients with idiopathic pulmonary fibrosis.

BACKGROUND: We investigated the expression of the IL-1 receptor on lung fibroblasts in patients with idiopathic pulmonary fibrosis (IPF) the histology of which was diagnosed as usual interstitial pneumonia. METHODS: 125I-labeled IL-1alpha bound to lung fibroblasts in a specific and saturable manner in lung fibroblasts obtained from both IPF patients and control subjects. RESULTS: Scatchard plot analysis revealed a single type of binding site in both groups. In total, 8,600 +/- 1,450 (mean +/-SD) sites/cell with a dissociation constant (Kd) of 1.83 +/- 0.28 (mean +/-SD) nM in patients with IPF and 7,920 +/- 1,470 sites/cell with a Kd of 2.43 +/- 0.49 nM in control subjects were counted. Furthermore, affinity cross-linking experiments revealed that the IL-1 receptor on lung fibroblasts in patients with IPF and control subjects had a molecular size of 80 kD. CONCLUSION: In conclusion, type I receptor was present on human lung fibroblasts, but there were no differences in number or affinity between IPF patients and controls.

Aged↗

PubMed1999

Proinflammatory or regulatory cytokines released from BALF macrophages of healthy smokers.

BACKGROUND: Chronic smoking influences bronchoalveolar lavage fluid (BALF) cell profiles in healthy subjects, which may alter profiles of inflammatory and regulatory cytokines. OBJECTIVE: We focused on the evaluation of smoking-related changes in the amounts of cytokines released from BALF macrophages. METHODS: We measured the amounts of immunoreactive culture supernatants by using ELISA. RESULTS: The amounts of IL-1 receptor antagonist (IL-1ra) were lower in smokers [n = 10, median 22.1 ng/ml, 25th and 75th percentiles (18.7-39.1)] than in nonsmokers [n = 10, 48.6 (39.2-66.1), p = 0.010]. In smokers, lipopolysaccharide stimulation revealed decreases in the amounts of interleukin-6 (IL-6) [nonsmokers: 2.1 ng/ml (0.68-5.4 vs. smokers: 0.5 (0.03-0.87), p = 0.049] as well as IL-1ra [nonsmokers: 69.2 ng/ml (48.3-83.8) vs. smokers: 27.3 (17.2-56.7), p = 0.028]. A delay in release from intracellular storage was not the cause of the reduced amounts of IL-1ra. In addition, interleukin-1beta (IL-1beta) was positively correlated with IL-6 and granulocyte macrophage colony-stimulating factor in nonsmokers, but not in smokers. Furthermore, the decreases in IL-1ra and interleukin-8 were correlated with the increase in the number of BALF macrophages in smokers, but not in nonsmokers. CONCLUSIONS: Chronic smoking caused changes in the profiles of cytokines released from BALF macrophages in healthy subjects. Decreases in the amounts of regulatory cytokines, but not prominent changes in the amounts of inflammatory ones, were characteristic.

Adult↗

PubMed1999

Monocyte chemotactic factors released from type II pneumocyte-like cells in response to TNF-alpha and IL-1alpha.

It has been reported that tumour necrosis factor (TNF)-alpha and interleukin (IL)-1 induce the release of monocyte chemotactic factors (MCF), including chemokines, from A549 cells, an alveolar type II cell line. However, the relative contribution of these chemokines to MCF is still uncertain. In the present study, the relative contribution of various chemokines released from A549 cells acting as MCF upon stimulation by TNF-alpha and IL-1alpha, was evaluated. TNF-alpha and IL-1alpha induced the release of MCF in a dose- and time-dependent manner (p<0.001). The release of MCF was inhibited by cycloheximide and lipoxygenase inhibitors. Molecular sieve column chromatography revealed multiple peaks of MCF (near 60 kDa, 25-22 kDa, 15-13 kDa, 8 kDa, and 400 Da). Leukotriene B4 (LTB4) receptor-antagonists inhibited MCF by 50% after 24 h and 30% after 72 h. Monocyte chemoattractant protein-1 (MCP-1), transforming growth factor (TGF)-beta, "regulated on activation, normal T-cells, expressed and secreted" (RANTES), and granulocyte-macrophage colony- stimulating factor (GM-CSF) were released significantly in response to IL-1alpha and TNF-alpha, and antibodies to MCP-1, GM-CSF, and RANTES inhibited MCF activity by 40, 5 and 20% after 24 h, and by 50, 20, and 10% after 72 h, respectively. Each antibody or LTB4 receptor-antagonist inhibited the corresponding column chromatography-separated molecular weight peak of MCF. These data suggest that A549 cells release monocyte chemoattractant protein-1 as the predominant monocyte chemotactic factor rather than granulocyte-macrophage colony-stimulating factor, RANTES, and transforming growth factor-beta, and that leukotriene B4 is constitutively released as a monocyte chemotactic factor.

Cell Line↗

PubMed1999

Different inhibitory effects of 5-S-glutathionyl-beta-alanyl-L-dopa (5-S-GA-L-D) analogues on autophosphorylation and substrate phosphorylation of Src protein tyrosine kinase.

Starting with 5-S-glutathionyl-beta-alanyl-L-dopa (1) and 5-S-glutathionyl-beta-alanyl-dopamine (2), a series of analogues with truncated glutathionyl and beta-alanyl-dopa moieties were synthesized, and their inhibitory effects on autophosphorylation and substrate phosphorylation reaction by c-Src and by epidermal growth factor receptor (EGFR) were evaluated. When the glutamyl residue was removed, the inhibitory effects on v-Src autophosphorylation decreased about 4- to 5-fold, and concomitant removal of the glutamyl and beta-alanyl residues resulted in a 40- to 60-fold decrease in the inhibition of v-Src autophosphorylation. On the other hand, these modifications had little effect on the inhibitory activity of substrate (Raytide) phosphorylation by c-Src. Interestingly, 5-S-cysteinyl dopamine inhibited the Src substrate phosphorylation reaction with comparable potency to that of genistein. Nonpeptide lipophilic derivatives had a similar inhibition on v-Src autophosphorylation but decreased inhibitory effects on substrate phosphorylation when compared to the lead compounds. Most compounds showed little effect on substrate phosphorylation by EGFR.

Dopamine↗

PubMed1999

Inhibition effects of 5-S-glutathionyl-N-beta-alanyl-L-dopa analogues against Src protein tyrosine kinase.

Twelve analogues of the antibacterial phenolic peptide 5-S-glutathionyl-N-beta-alanyl-L-dopa (5-S-GA-L-D, 1) were synthesized via orthoquinone using tyrosinase. Several synthesized compounds inhibited the v-Src autophosphorylation tyrosine kinase reaction with an IC50 value comparable to that of herbimycin A. The inhibition of c-Src substrate phosphorylation was much less active than v-Src autophosphorylation inhibition. 5-S-GA-L-D (1) and its analogous competed with peptide substrate and non-compared with ATP. The analogues showed no effects on substrate phosphorylation by epidermal growth factor receptor (EGFR), and this selectivity is the most characteristic feature of the 5-S-GA-L-D and its analogues (1-12).

3T3 Cells↗

PubMed1999

[Sulfonylureas and daily blood glucose profiles in elderly diabetics].

The aim of this study is to clarify the relationship between the efficacy of sulfonylureas and duration of diabetes in elderly diabetics. Daily blood glucose profiles were measured in 87 Type 2 elderly diabetic patients on sulfonylureas (tolbutamide, gliclazide or glibenclamide). Plasma glucose concentrations were determined at 08.00 (before breakfast), 10.00, 12.00 (before lunch), 14.00, 18.00 (before dinner), 20.00, 24.00, 03.00, 06.00, 08.00 hours. The subjects were divided into 4 sub-groups, according to their duration of the diabetes (< 10, 10-14, 15-19, 20 or more years). Mean plasma glucose values at 08.00, 10.00, 20.00, 03.00 and 06.00 hours were not significantly different among the four groups. However, mean plasma glucose values at 12.00, 14.00, 18.00, 00.00 hours and mean total blood glucose area under the daily profile (total BG) were significantly different among the four groups and the values in patients with a history of diabetes of 15 years or more increased. Duration of diabetes positively correlated with blood glucose values at 12.00, 14.00, 18.00, 00.00, 03.00 hours and total BG, and the dose of sulfonylureas positively correlated with blood glucose values at 12.00, 14.00, 18.00, 00.00 hours and total BG in multiple regression analysis. These results suggest that duration of diabetes and dose of sulfonylureas are important determinants of blood glucose control with sulfonylureas in elderly diabetic patients.

Aged↗

PubMed1999

[Pulmonary lesion associated with collagen vascular diseases: interstitial pneumonia].

Various pulmonary lesions can be found in patients with collagen vascular diseases. Not only the diseases but also treatment with various drugs relate to an occurrence of the pulmonary lesions such as infection, drug reaction, bronchiolitis and interstitial pneumonias. We focused on interstitial pneumonias based on our own experiences with patients whose lesions were diagnosed by surgical lung biopsy. Incidence of the interstitial pneumonias and diagnostic problems before biopsy using HRCT and BAL were evaluated. Comparison of interstitial pneumonias between idiopathic and associated with collagen vascular diseases is a problem to be solved.

Bronchoalveolar Lavage Fluid↗

PubMed1999

Angiotensin II receptor on BALF macrophages from Japanese patients with active sarcoidosis.

BACKGROUND: Serum angiotensin converting enzyme (ACE) activity and its changes are important markers for disease activity in patients with sarcoidosis. We earlier reported that ACE and its enzymatic product, angiotensin II (A-II), might play a role in maintaining macrophage/T lymphocyte alveolitis by enhancing an accessory function of macrophages in chronic active cases with sarcoidosis. AIM OF THE WORK AND METHODS: We examined whether A-II receptor is present on BALF macrophages by a receptor binding assay using 125I-labeled A-II, and by amplification of A-II receptor gene transcripts using RT-PCR methods and quantification of the amounts of transcripts by HPLC. RESULTS: The receptor binding assay suggested that specific binding of A-II to BALF macrophages was somewhat more prevalent in active sarcoidosis than in controls. A specific band for A-II receptor was detected by RT-PCR. A-II receptor gene expression was standardized as the ratio to beta-actin. An increased ratio was shown in active sarcoidosis (0.70 +/- 0.19; n = 14) compared to inactive cases (0.14 +/- 0.06; n = 5; p = 0.023) and healthy subjects (0.15 +/- 0.09; n = 5, p = 0.029). The ratio correlated positively with the percentage of BALF T lymphocytes (r = 0.60, p = 0.0056), and negatively with BALF macrophages (r = 0.60, p = 0.0055). No difference was detected between nonsmokers and smokers. CONCLUSION: The amounts of A-II receptor gene expression of BALF macrophages correlated with disease activity in patients with pulmonary sarcoidosis.

Adult↗

PubMed1999

[Anesthetic management for newborn pharyngeal teratoma].

Epignathus (pharyngeal teratoma) is a rare disease of newborns associated with a high mortality secondary to airway obstruction in the neonatal period. We report anesthetic management of a newborn with epignathus who underwent tumor resection. He was delivered vaginally at 39 weeks of gestation and Apgar scores were 9 at 1 and 5 min. The tumor originated from the palate, almost filled the oral cavity and protruded through the mouth with its external part 6 x 7 cm in size. He could breathe with the head and mass turned to the left. The excision of the tumor was scheduled on the fifth day of life. Mask ventilation and laryngoscopy were considered impossible. Fiberoptic nasal intubation was successfully performed with topical anesthesia without sedation. Tumor was resected with blood loss of 103 gm. The trachea was extubated on the third postoperative day and the postoperative course was uneventful. For safe management of cases of pharyngeal teratoma, careful preoperative assessment of the airway is most important and sufficient preparation and careful intubation are mandatory to keep airway patent. The perioperative bleeding from the tumor and the airway obstruction by the tumor or its remnant after the excision could also be hazardous to the airway.

Anesthesia, Inhalation↗

PubMed1999

[Bronchiolitis obliterans organizing pneumonia].

A clinicopathological disease entity of bronchiolitis obliterans organizing pneumonia (BOOP) is reviewed. Various clinical backgrounds can be found in patients who were histopathologically diagnosed as BOOP. Historically, the idiopathic BOOP has been identified as a clinicopathological disease entity in 1985. BOOP shows clinically interstitial processes, however some confusion is still present in the aspect of differentiation with bronchiolitis obliterans (BO). Histopathologically, BOOP shows organizing reaction to subacute lung injuries, and absence of remodeling separates BOOP from other chronic fibrosing processes such as UIP. Clinically, there is differences in clinical outcomes between idiopathic, and secondary BOOP. We reviewed whether BOOP can be classified as interstitial pneumonia.

Cryptogenic Organizing Pneumonia↗

PubMed1999

Hospital-based historical cohort study of 234 histologically proven Japanese patients with IPF.

BACKGROUND AND AIM OF THE WORK: Variable clinical courses have been recognized in patients with idiopathic pulmonary fibrosis (IPF), but the disease is known to have a fundamentally unfavorable outcome. In addition, we have yet to fully understand whether the regional or population differences are negligible or not. Therefore, we analyzed the clinical outcomes in Japanese IPF patients in a hospital-based historical cohort study. METHODS: A questionnaire was used to collect records for retrospective analysis of the IPF patients. We analyzed data collected from 51 hospitals and clinics in Japan on 234 Japanese IPF patients (males 170, females 64) with IPF who had been followed up after diagnosis. RESULTS: 1. Five and ten years after the detection of exertional dyspnea, the overall survival rates were 41.0% and 20.1%, respectively. A higher age at the time of detection (over 59) was related to a decrease in survival rates. 2. The incidence of complications was no higher in the corticosteroid-treated cases than in the untreated cases. The effects of corticosteroid treatment on survival was not confirmed in this type of study. CONCLUSIONS: The survival rates of histologically proven Japanese patients with IPF were similar to the rates previously reported in different populations. Age at the detection of exertional dyspnea was critical in terms of the survival rate.

Adrenal Cortex Hormones↗

PubMed1999

Bone mineral density and vitamin D in patients with sarcoidosis.

BACKGROUND AND AIM OF THE WORK: Imbalance of calcium homeostasis has ben frequently detected in sarcoidosis patients. In our previous study abnormal metabolism of calcium was detected in about 40% of the patients. In the current study, bone mineral density (BMD) was investigated in relation to serum concentration of calcium and vitamin D. METHODS: Consecutive groups of fifteen men (40 +/- 12 years, as mean +/- SD) and 18 women (51 +/- 11 years) with histologically proven sarcoidosis who had not been treated with corticosteroids prior to the study were studied together with control groups. BMD was measured with dual-energy X-ray absorptiometry (DXA) of lumbar vertebrae, and the Z score was calculated to avoid bias of sex and age. Serum concentrations of ionized calcium (s Ca2+), 1,25-Dihydroxyvitamin D (s1, 25(OH)2D3) and osteocalcin (sOC) were measured at the same time. RESULTS: In women with sarcoidosis only, sCa2+ and s1,25(OH)2D3 showed inverse correlations with Z score of BMD (r = 0.482, p < 0.05 and r = -0.635, p < 0.01, respectively), and s1,25(OH)2D3 showed a positive correlation with sOC (r = 0.658, p < 0.01). Multiple regression analysis showed that age and s1,25(OH)2D3 were the contributory factors to BMD in female patients. Such a significant correlation was not observed in men with sarcoidosis or controls. CONCLUSIONS: 1,25(OH)2D3 seemed to be contributory to accelerated turnover of the bone and reduction of BMD in women with sarcoidosis.

Absorptiometry, Photon↗

PubMed1999

[Docetaxel as a single agent in cases of advanced/recurrent breast cancer previously treated with anthracycline].

Docetaxel 60 mg/m2 was administered via 1-hour intravenous infusion every 3-4 weeks to 20 patients with advanced/recurrent breast cancer who had previously received anthracycline therapy. The overall response rate was 30.0% (6/20), with 1 complete remission and 5 partial remissions. The median response duration was 120 days. Response rates classified by recurrence site were soft tissues 43.0%, liver 33.3%, lung 33.3%, and bone 23.1%, respectively. Stable disease (prolonged NC) which continued more than 6 months was observed in patients with bone metastasis. Grade 3-4 leukocytopenia and neutropenia were observed in 45.0% and 55.0%, respectively. Grade 3-4 leukocytopenia and neutropenia were frequently seen in patients who had received anthracycline of more than 500 mg/body as first-line chemotherapy. Docetaxel thus seems to be effective in patients with advanced/recurrent breast cancer who are refractory to anthracycline therapy.

Adult↗

PubMed1998

An avian pathogenic Escherichia coli strain produces a hemolysin, the expression of which is dependent on cyclic AMP receptor protein gene function.

An avian pathogenic Escherichia coli strain M1000 showed a clear zone of erythrocyte lysis on sheep blood agar plates. The hemolytic activity was not detected in the culture supernatant nor was any DNA sequence homologous to the E. coli alpha-hemolysin gene detected in the chromosome or plasmid DNA of the strain, indicating that the observed hemolysis was different from alpha-type. To identify the genetic determinant responsible for the hemolysis, we performed random Tn5 insertional mutagenesis and obtained one mutant, named M5005, that totally lacked the hemolytic activity. Cloning and nucleotide sequencing of the region flanking the transposon insertion site in the M5005 chromosome revealed that the transposon was inserted within an open reading frame of the cyclic AMP receptor protein (CRP) gene, which is involved in one of the global regulatory networks of gene expression in E. coli. Nucleotide sequence analysis of the intact crp gene of the strain M1000 showed that the CRP protein of M1000 is 99% identical to that of K-12. Introduction of the intact crp gene on a low copy plasmid into the mutant M5005 restored the hemolytic phenotype, confirming that the mutation site in M5005 was in the crp gene. CRP plays a central role in catabolite repression, the phenomenon by which the synthesis of many enzymes required to metabolize various sugars is repressed in the presence of glucose. When the hemolytic activity of E. coli M1000 grown in the presence of glucose was examined, the hemolysis was totally impaired. These results indicate that the avian pathogenic E. coli strain M1000 produces a hemolysin the expression of which is dependent on crp gene function.

Amino Acid Sequence↗

PubMed1998

Micellar electrokinetic chromatography as an alternative to high-performance liquid chromatography for separation and determination of phenolic compounds in Japanese spirituous liquor.

A capillary electrophoretic method has been developed for the separation and determination of vanillin, ferulic acid, vanillic acid and 4-vinylguaiacol in Japanese spirituous liquor. These phenolic compounds were extracted from Japanese spirituous liquor by solid-phase extractioin and were successfully separated by micellar electrokinetic chromatography (MEKC) with 25 mM sodium dodecyl sulfate. MEKC required short analysis times and it could be an alternative technique for the analysis of vanillin, ferulic acid, vanillic acid and 4-vinylguaiacol in Japanese spirituous liquor. These phenolic compounds in five Japanese spirituous liquor samples were determined by this method. No significant difference in separation patterns and amounts was observed among the repeated analysis.

Alcoholic Beverages↗

PubMed1998

Simultaneous measurement of alpha-amylase and glucoamylase activities in sake rice koji by capillary electrophoresis of sodium dodecyl sulfate-protein complexes and activity measurement of glucoamylase by in-capillary enzyme reaction method.

Capillary electrophoresis (CE) of sodium dodecyl sulfate (SDS)-protein complexes using a nongel sieving matrix (CE-SDS) has been applied to the simultaneous analysis of alpha-amylase and glucoamylase activity in sake rice koji which is employed for the brewing of sake. Alpha-amylase and glucoamylase in sake rice koji extracts were successfully analyzed by CE-SDS. Alpha-amylase and glucoamylase were found to have molecular masses of 53000 and 63000 Da, respectively, as determined by the migration times of eight standard proteins. These values agree with those determined by SDS-polyacrylamide gel electrophoresis (PAGE). The results of CE-SDS method were compared with those achieved by the official method. The relative standard deviations (RSD) of the alpha-amylase and glucoamylase activities by CE-SDS were less than 5.0% in both intra-day and inter-day experiments. An electrophoretic analysis of products of an enzyme reaction of a substrate by in-capillary reaction was also useful for the activity measurement of glucoamylase in sake rice koji. p-Nitrophenyl-beta-D-maltoside (PNP-Mal) was employed as a substrate and p-nitrophenyl-beta-D-glucopyranoside (PNP-Glu) was the product of the enzyme reaction. The glucoamylase activity of sake rice koji samples gave the good linear relationship with the peak area observed in the in-capillary enzyme reaction method. The glucoamylase activity in sake rice koji was measured by either CE-SDS or the in-capillary enzyme reaction more easily than by the official method. Both methods can be applied to the routine quality control of alpha-amylase and glucoamylase activities in sake rice koji.

Alcoholic Beverages↗

PubMed1998

The value of long-term clinical follow-up for cases of spontaneous carotid cavernous fistula.

To clarify the value of clinical long-term follow-up with radiological examination, ranging from 12 to 63 months (average: 35 months), 18 consecutive patients suffering from spontaneous carotid cavernous fistula (CCF), were studied prospectively. Five aged patients without aggressive symptoms were treated conservatively, and the other 13 underwent transarterial embolization. The radiological follow-up was primarily by magnetic resonance angiography (MRA), performed from 2 to 6 times (average: 4.1 times) during the follow-up period. In three cases, CCFs persisted, but the other fifteen (83%) demonstrated complete cure as defined by long-term follow-up MRA. The three patients with persistent CCFs were comparatively young, less than 60 years old, had no atherosclerotic factors and demonstrated multiple venous drainage routes with cortical venous drainage on angiography. In two of them, the symptoms completely disappeared, and the other had only mild chemosis. However, surprisingly, in two, MRA revealed residual CCF with drainage into only cortical veins through the sphenoparietal sinus, this radiological finding being well known to signify danger. During the follow-up period, central retinal vein thrombosis occurred in two cases. The common point in these cases was that the superior ophthalmic vein was the only venous drainage route. This is also a point requiring care. We therefore emphasize the importance of careful long-term radiological follow-up for spontaneous CCF patients even when their symptoms improve or disappear. MRA is particularly suitable for this purpose and applicable in the out-patient clinic because of its non-invasive nature.

Adult↗