Biomedical subjects

M Fujino

Publications and source records attributed to M Fujino.

At least 109 records · Page 6Linked to original sources

PubMed1997

Structure and function of the hemolymph node in rats.

Hemolymph nodes (HLs) are unique lymph nodes, in that their lymphatic sinuses contain numerous erythrocytes. In this study, we compared the internal structure and immunologic function of HLs with those of ordinary lymph nodes (OLs) and the spleen. Electron microscopy revealed erythrocytes passing through the walls of blood vessels in the intermediate sinus area (IMSA) of a HL between expanded endothelial cell junctions. However, no direct communication was found between lymphatic sinuses and blood vessels. Numerous carbon particles appeared in the IMSA of HLs on 5 days after intravenous carbon particle injection, while OLs lacked particle deposition. Immunohistochemical studies showed that lipopolysaccharide (LPS) reached the IMSA of HLs and extravasated into medullary cords 4 hours after intravenous LPS injection, resulting in the appearance of more IgM-stained lymphocytes in the IMSA of HLs than in that of OLs on day 5. The ability of organs to produce antibodies was determined by counting the number of plaque forming colonies after intravenous injection of sheep red blood cells (SRBC). The HLs antibody-producing ability was between that of OLs and the spleen. These results suggest that HLs possess functionally open blood vessels in the IMSA and their immunologic capability is between that of OLs and the spleen. These findings suggests that HLs are lymphoid organs that have characteristics between those of the OLs and the spleen, both ultrastructurally and functionally.

Animals↗

PubMed1997

[A case of primary endobronchial mucosa-associated lymphoid tissue lymphoma with middle lobe atelectasis].

A 60-year-old woman was admitted for evaluation of an abnormal shadow in her chest X-ray. A chest roentgenogram revealed middle lobe atelectasis and a tomogram showed masses mainly in the right intermediate bronchus. In bronchoscopy, the bronchus was severely narrowed by irregularly surfaced masses. Microscopic examinations of transbronchial biopsy specimen showed the mucosal and submucosal tissue diffusedly infiltrated mainly by lymphocytes composed of small lymphocytes, centrocyte-like cells and monocytoid cells. The B-cell origin was suggested by a positive L-26 stain. Monoclonality of lymphocytes was proven by Southern blot analysis and in situ hybridization. The diagnosis was primary endobronchial lymphoma of mucosa-associated lymphoid tissue. The case was treated with chemotherapy protocol (CAMBO-VIP) and has been in complete remission for 20 months.

Antineoplastic Combined Chemotherapy Protocols↗

PubMed1996

Effects of a new endothelin receptor antagonist, TAK-044, on myocardial stunning in dogs.

The effects of a new endothelin receptor antagonist, TAK-044, (cyclo[D-alpha-aspartyl-3-[(4-phenylpiperazin-1-yl)carbonyl]L-alan yl-L-alpha aspartyl-D-2-(2-thienyl)glycyl-L-leucyl-D-tryptophyl]disodium salt, on ischemic and post-ischemic myocardial dysfunction (stunned myocardium) were studied in anesthetized open-chest dogs. A short (15 min) occlusion of the left anterior descending coronary artery followed by 5-h reperfusion significantly reduced myocardial segment shortening during and after the ischemic period in the ischemic region. Regional myocardial blood flow was also decreased significantly 10 min after the occlusion, whereas it returned almost completely to its pre-ischemic value 5 h after reperfusion TAK-044 (3 mg/kg,i.v.) administered 10 min before occlusion significantly improved the reduced myocardial segment shortening in the ischemic region during and after occlusion. Cardiovascular hemodynamics and regional myocardial blood flow in a TAK-044-treated group were identical to those in the control group. These results indicate that endogenous endothelin contributes to the cause of ischemic and post-ischemic myocardial dysfunction without changing either hemodynamics or regional myocardial blood flow.

Animals↗

PubMed1996

Pharmacological characterization of cardiovascular responses induced by endothelin-1 in the perfused rat heart.

The effects of the endothelin receptor antagonist TAK-044 (cyclo[D-alpha-aspartyl-3-[(4-phenylpiperazin-1-yl)carbonyl]-L-ala nyl-L-alpha-aspartyl-D-2-(2-thienyl)glycyl-L-leucyl-D-tryptophyl]+ ++disodiu m salt) and BQ-123 (cyclo[D-Asp-Pro-D-Val-Leu-D-Trp]) were studied in the rat heart to characterize the receptor subtypes responsible for the cardiovascular actions of endothelin-1. Endothelin-1 induced a transient decrease and subsequent increase in perfusion pressure in perfused rat hearts, and increased left ventricular developed pressure. TAK-044 diminished these endothelin-1-induced responses (100 pmol/heart) with IC50 values of 140, 57 and 1.3 nM, respectively. BQ-123 (1-30 mu M) partially inhibited the endothelin-1-induced hypertension (30-40%) in the rat heart, and failed to inhibit the hypotension. The positive inotropic effect of endothelin-1 was abolished by BQ-123. Neither indomethacin (10 mu M) nor Nomega-nitro-L-arginine methyl ester (100 mu M) attenuated the endothelin-1-induced hypotension. TAK-044 and BQ-123 attenuated the positive inotropic effect of endothelin-1 in rat papillary muscles. In rat cardiac membrane fractions, TAK-044 and BQ-123 inhibited [125I]endothelin-1 binding to endothelin ET(A) receptors with IC50 values of 0.39 +/- 0.6 and 36 +/- 9 nM, respectively, whereas only TAK-044 potently blocked the endothelin ET(B) receptor subtype (IC50 value: 370 +/- 180 nM). These results suggest that endothelin-1 modulates cardiovascular functions in the rat heart by activating both endothelin ET(A) and endothelin ET(B) receptors, all of which are sensitive to TAK-044.

Analysis of Variance↗

PubMed1996

Cloning and expression of the canine interferon-beta gene.

We isolated the canine interferon-beta (IFN-beta) gene from dog liver chromosomal DNA by the polymerase chain reaction (PCR). The coding region encodes a predicted protein of 197 amino acids, consisting of a signal sequence of 32 amino acids and a mature IFN-beta of 165 amino acids. In the IFN-beta sequence, there are five potential N-glycosylation sites and four cysteine residues. Canine IFN-beta has 44% and 60% amino acid sequence homology with murine and human IFN-beta, respectively, whereas it has only 28% homology with canine IFN-alpha. The canine IFN-beta gene was expressed in insect cells under the control of the polyhedrin promoter in a recombinant baculovirus. After infecting Sf21 cells with the recombinant baculovirus, IFN activity was detected in the culture medium, indicating that it is secreted from the cells. This activity was stable from pH 2 to 12 for 18 h at 4 degrees C. Southern blot analysis indicated that the gene for canine IFN-beta is a single gene in the dog haploid chromosome.

Amino Acid Sequence↗

PubMed1996

Kinetics of monocytoid B lymphocytes in localized inflammatory lesions induced by lipopolysaccharide in mice.

Immunohistochemistry was used to study the kinetics of B lymphocytes (B-lys) in the early stages of the localized inflammatory response induced in SMA mice by the subcutaneous injection of lipopolysaccharide (LPS). At the injection sites, medium-sized B-lys formed early inflammatory lesions with neutrophils and activated macrophages on days 1 and 2. The B-lys were morphologically similar to monocytes, but were not stained with Mac1 antibody. Remarkably the B-lys showed the phenotypes of B220+, IgM+, IgD (slight to negative), Ly-1- and CD23- by double immunohistochemical staining. The B-lys were also positive for alkaline phosphatase. Consequently the B-lys could be identified as monocytoid B-lys or marginal zone B-lys. Plasmacytic B-lys and plasma cells were first observed on days 3 and 4, but no lymphoid follicles were found at the injection sites. In the inguinal lymph nodes, the same B-lys responses were mainly induced in the paracortical lesions (T cell areas) preceding the formation of activated germinal centers (GC). These findings suggested that the B-lys, induced by injections of LPS, matured into plasma cells in the localized inflammatory lesions independent of GC, and that they were different from follicular B-lys.

Animals↗

PubMed1996

[Intralobar pulmonary sequestration associated with asymptomatic aspergillosis: a case report].

One case of intralobar pulmonary sequestration associated with asymptomatic aspergillosis are reported. A 39-year-old woman was admitted to our hospital because of abnormal shadow in the right lower lobe detected on mass miniature chest X-ray. The patient had been asymptomatic, and an aortogram showed an anomalous artery originating from abdominal aorta and entering the abnormal shadow suggesting sequestration. The right S10 segmentectomy was done. With bacterial culture of fluid in the specimen and on microscopic investigation this proved to be an intralobar pulmonary sequestration infected with Aspergillus. This case shows the necessity of prompt resection of intralobar pulmonary sequestration, even if asymptomatic.

Adult↗

PubMed1996

[Serum concentrations of the pyridoxal and pyridoxal-5'-phosphate in children during sustained-release theophylline therapy].

This study was undertaken to investigate the effect of sustained-release theophylline preparations on circulating vitamin B6 concentrations in children with asthma. Twenty-six children with asthma were treated with sustained-release theophylline preparations. Serum concentrations of theophylline and vitamin B6 [pyridoxal (PL) and pyridoxal-5'-phosphate (PLP)] were evaluated. The results obtained from this study were as follows: 1) The serum PL was significantly correlated with the duration of theophylline administration (r = -0.42). 2) The serum PL and PLP concentrations were significantly correlated with serum theophylline concentrations (r = -0.47, r = -0.59, respectively). 3) Serum PL and PLP concentrations in children treated for more than 5 weeks were significantly lower than those treated for less than 4 weeks (p < 0.01, p < 0.05, respectively). 4) Serum PL and PLP concentrations in children within theophylline therapeutic ranges (5-15 micrograms/ ml) were significantly lower than those with less than 5 micrograms/ml (p < 0.01, p < 0.01, respectively). These findings suggest that long-term theophylline therapy or serum theophylline concentration within the therapeutic ranges (5-15 micrograms/ml) can depress serum vitamin B6 concentration in children with asthma. Theophylline-induced seizure may be caused by decrease in gamma-aminobutyric acid concentration in the brain result from decrease in vitamin B6 concentration, even if the serum theophylline concentration is within the therapeutic ranges.

Asthma↗

PubMed1995

Prognostic significance of p53 and ras p21 expression in nonsmall cell lung cancer.

BACKGROUND: Alterations of the p53 gene are one of the most common genetic changes in various types of cancer, including lung cancer. Abnormalities in the ras genes, including point mutations and overexpression, are another common feature in the molecular biology of lung cancer and are associated with a poorer prognosis. The authors' purpose was to determine expression of the mutated p53 gene in nonsmall cell lung cancer (NSCLC) specimens that were studied for expression of ras p21 and to document whether altered p53 expression was also an important factor for survival. METHODS: Ninety-six patients with NSCLC underwent surgical resection between 1977 and 1985, 63 of whom received postoperative combination chemotherapy. None received radiation therapy. Tumor specimens were analyzed for altered p53 expression by immunohistochemistry. Univariate and multivariate analyses were performed to assess the association between p53 expression and survival. RESULTS: Fifty-six (58%) of 96 tumor specimens showed altered p53 expression, and 91 patients were analyzed for survival. Altered p53 expression did not correlate with clinicopathologic characteristics except for postsurgical pathologic tumor (pT) classification. The patients with altered p53 expression survived for a significantly shorter period after surgery than those without p53 expression, including all patients who underwent resection and potentially curative resection (P = 0.02 and P = 0.048, respectively, generalized Wilcoxon test). Multivariate analysis showed independent prognostic significance for altered p53 expression (hazard ratio [HR] = 1.72, P = 0.04) and surgical cure (HR = 4.69, P < 0.001). The combined analysis of mutated p53 and ras p21 expression in the same tumor specimens revealed that patients with p53- and ras p21-negative tumors survived the longest among those with different p53 and ras p21 features (P = 0.005, generalized Wilcoxon test). CONCLUSION: Altered p53 expression is a significant and independent negative prognostic factor for patients with surgically resected NSCLC: Combined immunohistochemical analysis of mutated p53 and ras p21 expression can divide patients with NSCLC into more accurate prognostic groups. If the current findings can be confirmed in larger prospective studies, combined immunohistochemical analysis of mutated p53 and ras p21 expression can be a useful clinical tool for stratifying patients with NSCLC into accurate prognostic groups and for identifying the population with a different risk of recurrence.

Aged↗

PubMed1995

Effects of a potent, non-selective endothelin receptor antagonist, [Thr18, gamma-MeLeu19]-endothelin-1, on the isolated blood vessels.

The endothelin (ET) receptor has been classified as the ETA1, ETA2, ETB1 and ETB2 subtypes. BQ-123 and RES-701-1 are the ETA1 and ETB1 antagonists, respectively, whereas BQ-788 is the ETB1/ETB2 antagonist. To find the ETA2 antagonist, the effects of [Thr18, gamma-MeLeu19]-ET-1 (TM-ET-1) were examined. In the rabbit saphenous vein, contraction induced by ET-1 is due to simultaneous activation of the ETA1, ETA2, ETB1 and ETB2 receptors whereas contraction induced by sarafotoxin S6c is mediated by the ETB1 and ETB2 receptors. Neither BQ-123 nor RES-701-1 inhibited these contractions. TM-ET-1 antagonized the effects of ET-1 and sarafotoxin S6c. In the vein in which the ETB1/ETB2 receptors were desensitized and the ETA1 receptor was inhibited by BQ-123, ET-1 still induced contraction. This contraction was inhibited by TM-ET-1. In the rat aorta with endothelium, ET-3 induced endothelium-dependent relaxation by activating the endothelial ETB1 receptor. TM-ET-1 inhibited the effect of ET-3 without changing the relaxation induced by carbachol. In the rat aorta without endothelium, ET-1 induced contraction by activating the smooth muscle ETA1 receptor. TM-ET-1 inhibited this contraction. These results suggest that TM-ET-1 is a novel antagonist against the ETA2 receptor that also inhibits other types of the ET receptors.

Animals↗

PubMed1995

Gene organization of chicken anemia virus.

The genomic DNA of chicken anemia virus (CAV) was cloned and sequenced from a Japanese isolate CAA82-2. The nucleotide sequence of CAA82-2 isolate was 98% identical with that of the European Cuxhaven-1 strain (Noteborn et al., J. Virol. 65, 3131-3139, 1991). Nine open reading frames (ORFs) consisting of more than 100 nucleotides were found, i.e., four ORFs (CA1-CA4) on the plus strand and five ORFs (CA1R-CA5R) on the minus strand. These ORFs with the exception of CA4 are conserved between the two CAV isolates. All of these ORFs were expressed in Escherichia coli as fusion proteins with beta-galactosidase. By Western blot analysis, the CA2 and CA3 fusion proteins were found to react with CAV-infected chicken sera. Rabbit hyperimmune sera against the CA1, CA2, and CA3 fusion proteins were produced and tested their reactivity to CAV-infected cells. Two viral proteins with the apparent size of 54 and 16 kDa reacted with the antibodies against CA1 and CA3 fusion proteins, respectively. The 16-kDa protein, CA3, was suggested to be a major immunogen on CAV infection.

Animals↗

PubMed1995

Mechanical and histological investigation of hydrothermally treated and untreated anodic titanium oxide films containing Ca and P.

In a previous study a new method for forming thin hydroxyapatite (HA) layers on titanium was described. Titanium was anodized at 350 V in an electrolyte solution containing sodium beta-glycerophosphate and calcium acetate, and an anodic titanium oxide film containing Ca and P (AOFCP) was formed on the surface. Then numerous HA crystals were precipitated on the AOFCP during hydrothermal treatment in high-pressure steam at 300 degrees C. In this study three types of hydrothermally treated films differing in amounts of precipitated HA crystals and tensile adhesive strength, and untreated films were histologically and mechanically investigated in a transcortical rabbit femoral model for 8 weeks of implantation using light microscopy, scanning electron microscopy (SEM), and push-out tests. Machined titanium and HA ceramics served as control materials. The push-out shear strength and bone apposition of the AOFCP significantly increased after hydrothermal treatment, and were equivalent to those of HA ceramics, although the HA layer on the AOFCP was thin at 1-2 microns. From SEM observation of the pushed-out specimen, it was found that the thin HA layer had directly bonded to bone but the AOFCP had not. The push-out strength of the hydrothermally treated film resulted from the chemical bonding of the bone-HA layer interface, while that of the untreated film resulted from mechanical interlocking force between bone and the microprojections. There was a small difference in bone apposition but no significant difference in push-out strength with the amount of precipitated HA crystals on the treated films. Among the treated films, the film formed at the lowest electrolyte concentration showed the lowest bone apposition because of incomplete covering by the HA crystals, and showed the highest stability against mechanical failure because the adhesive strength was very high at about 38 mPa. Also, the hydrothermally untreated anodic oxide films, whose surfaces were rough as a result of the large microprojections, showed much higher push-out strength and bone apposition than titanium. The good hard-tissue compatibility may be attributed to the surface roughness and the possible inhibition of titanium ion release from the specimen.

Animals↗

PubMed1995

Human papillomavirus type 18 DNA and E6-E7 mRNA are detected in squamous cell carcinoma and adenocarcinoma of the lung.

To provide an accurate evaluation of the association of human papillomavirus (HPV) with lung cancer, 36 cases of lung cancer were analysed for HPV DNAs by polymerase chain reaction (PCR) with dot-blot and Southern blot analyses, and for the transcripts from the E6-E7 transforming region by in situ hybridisation (ISH). HPV-18 DNA was detected in three (8%) of 36 specimens; histologically, in one (10%) of 10 squamous cell carcinomas and two (9%) of 22 adenocarcinomas. Neither HPV-16 nor -33 DNA was detected in any cases examined. Expression of E6-E7 mRNA was confirmed in the cases which contained, HPV-18 DNA. HPV-18 may play an important role in the development and progression of cancer in some cases of both squamous cell carcinoma and adenocarcinoma of the lung.

Adenocarcinoma↗

PubMed1995

Simultaneous use of the PCR-SSCP method and immunohistochemistry for increasing the detection efficacy of p53 abnormalities in human lung cancer.

Malignant neoplasms possess multiple genetic abnormalities. Among those, p53 gene abnormalities are the most frequent in human neoplasms. To screen for p53 abnormalities, both immunohistochemistry (IHC) and the polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) methods are commonly used, but neither can detect specimens simultaneously by the two methods, 12 abnormal cases (34%) were detected by IHC and 9 abnormal cases (26%) were detected by PCR-SSCP. Six abnormal (17%) and 20 normal (57%) cases showed concordant results between the methods, although 9 cases (26%) showed discordance, including 6 IHC-positive cases (17%) and 3 SSCP-positive cases (9%). Because it is known that some p53 abnormalities are detected only by IHC or only by PCR-SSCP, discordant cases should be assessed as possessing abnormalities. Fifteen cases (43%) were finally assessed as abnormal. These results suggest that the two methods together can increase the sensitivity of screening for p53 abnormalities.

Gene Expression↗

PubMed1995

Pharmacology of a non-selective ETA and ETB receptor antagonist, TAK-044 and the inhibition of myocardial infarct size in rats.

1. The aims of the present study were to characterize the pharmacological profile of a new endothelin (ET) receptor antagonist, TAK-044 and to consider whether it limits the extension of myocardial infarct size in rats. 2. Binding of [125I]-ET-1 to ET receptors on rabbit ventricular and cerebellar membrane fractions was inhibited by TAK-044 with IC50 values of 3.8 nM and 130 nM, respectively. 3. It inhibited ET-1, ET-2 and ET-3-induced vasoconstriction of porcine isolated coronary arteries in a competitive (ET-1, ET-2) and a non-competitive (ET-3) manner. 4. In the rat in vivo, the ET-1-induced blood pressure changes including transient hypotension followed by sustained hypertension, were inhibited by TAK-044 (0.1-10 mg kg-1, i.v.) in a dose-dependent manner. 5. Acute myocardial infarction induced by 1 h coronary occlusion followed by 24 h reperfusion in rats caused an infarct size of 60 +/- 2% (n = 12) of the area-at-risk by weight. 6. Intravenous injection of TAK-044 10 min before coronary occlusion reduced the infarct size in a dose-dependent manner: 32% and 54% reductions at 1 and 3 mg kg-1, respectively. 7. TAK-044 administered 10 min before or 1 h after reperfusion (1 mg kg-1, i.v.) showed similar inhibitory effects: 34% and 23% reductions, respectively. 8. We conclude that TAK-044 is an ETA/ETB receptor antagonist which shows strong inhibitory effects on the extension of myocardial infarct size after coronary artery occlusion-reperfusion in rats.

Animals↗

PubMed1995

PACAP stimulates catecholamine release from adrenal medulla: a novel noncholinergic secretagogue.

The secretory response of the adrenal medulla to pituitary adenylate cyclase-activating polypeptide (PACAP), a novel polypeptide with 68% structural homology to vasoactive intestinal polypeptide (VIP), was investigated in anesthetized rats by in vivo microdialysis. The injection of PACAP (1.5 nmol) caused a greater amount of increase in catecholamine concentration than carbachol (30 nmol) or VIP (30 nmol) in the dialysate for a period of 60 min. The ratios of norepinephrine to epinephrine in the dialysate after stimulation with these compounds showed no change from resting conditions. The simultaneous application of both nicotinic and muscarinic antagonists (10 mM mecamylamine and 1 mM atropine) eliminated the increase in catecholamine secretion induced by carbachol. In contrast, PACAP-induced catecholamine secretion was not inhibited by these cholinergic antagonists. These data led to the following conclusions: 1) PACAP was more potent than either carbachol or VIP in enhancing the secretion of catecholamine from the adrenal medulla, and 2) PACAP-induced catecholamine secretion was due to the direct action of PACAP on the adrenal medulla rather than an indirect action mediated by acetylcholine release. These results strongly suggest that PACAP is a noncholinergic secretagogue of the adrenal medulla in rats.

Adrenal Medulla↗

PubMed1995

Application of a unique automated synthesis system for solution-phase peptide synthesis.

An automated synthesis system, which is suitable for repetitive syntheses using similar reaction procedures, was used to synthesize systematically a library of all possible dipeptides (25) and tripeptides (125) from 5 protected amino acids. The apparatus has also been applied to the automated synthesis of 10 fragment tripeptide derivatives that are constituents of the hormone PACAP-27. The measured molecular optical rotation values of the library of 125 tripeptides were found to correlate well with calculated values obtained by summation of the molecular optical rotation values for the constituent amino acids.

Amino Acid Sequence↗

PubMed1995

An immunohistochemical study of the significance of a new 31.5-kD ouabain receptor protein isolated from cat cardiac muscle.

A new 31.5-kD ouabain receptor protein (NORP), which is independent of Na(+)-K+ ATPase, was recently isolated selectively from transverse tubule membrane-junctional sarcoplasmic reticulum (TTM-JSR) complexes of cat cardiac muscle. We investigated the role of this NORP in cardiac function with special reference to the positive inotropic effect (PIE) of ouabain, preparing and using a monoclonal antibody (MoAB, immunoglobulin) raised against the receptor protein. Electrically stimulated papillary muscles were immersed in a Tyrode solution containing the anti-NORP MoAB (40 microM), of which the binding potency was high enough for immunological use, for 60 min and then washed out. Thirty minutes after removal of the MoAB, both twitch and K-contracture were still inhibited, but both resting and action potentials and caffeine-induced contracture were unchanged, indicating that NORP plays a key role in excitation (E)-contraction (C) coupling. The intracellular localization of the protein was investigated by immunohistochemical electron microscopy, and the protein was shown to be located on the TTM, the location being probably its external surface and opposite to feet which occupy the TTM-JSR gap. These results indicate that E-C coupling of cardiac muscle cells is mediated through NORP and that ouabain-PIE occurs through the influence of ouabain on NORP in the E-C coupling process.

Animals↗