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Biomedical subjects

M Fujino

Publications and source records attributed to M Fujino.

At least 163 records · Page 9Linked to original sources

Identification of bone morphogenetic protein-2 in early Xenopus laevis embryos.

Polyclonal antibodies capable of reacting with amphibian bone morphogenetic protein (BMP-2 and -4) were raised in rabbits by immunization with a synthetic 21 amino acid peptide which corresponds to a sequence residing in the mature protein of Xenopus BMP-2 (xBMP-2). The antibodies recognized an embryonic BMP as well as mammalian and bacteria expressed recombinant xBMPs. The antibodies detected, under reducing conditions, a 30 kDa protein in the extract of oocytes and embryos during early development. Interestingly, acidification of the extract from each developmental stage yielded a protein band of smaller molecular weight of 18 kDa, which is similar in size to reduced form of mature BMPs purified from mammalian species. Two-dimensional electrophoresis employed to examine the molecular weight of unreduced forms using the antibody, revealed that both molecular forms are monomeric in the embryos. The result suggests that at least BMP-2 mRNA previously detected in early embryos, is translated into peptide but the dimerization may be incomplete or strictly limited in these embryos.

Amino Acid Sequence↗

[Study on local recurrence in patients undergoing curative surgical resection of lung cancer].

Clinical features of recurrence in patients of lung cancer undergoing curative surgical resection were examined with special reference to the local recurrence. Subjects were 308 patients, consisting of 160 adenocarcinomas, 121 squamous cell carcinomas, 15 large cell carcinomas, 12 small cell carcinomas. They underwent curative resection in our department between 1973 and 1987. Local recurrence developed in 54 patients (18%). There was no significant difference in the incidence of local recurrence among the four histological types of carcinoma. According to the pathological stage, the incidence was 9% in stage I, 15% in stage II, and 35% in stages IIIA+IIIB. The local recurrence was subdivided into the following patterns: 1) lymphatic metastases to the hilar, mediastinal, or supraclavicular sites, 2) recurrence at the surgical margin, 3) malignant pleuritis or pericarditis, 4) so-called "endobronchial metastasis". The incidence of recurrence according to the patterns was 15%, 2%, 2% and 1%, respectively. The incidence of recurrence due to lymphatic metastases was correlated significantly with the pN factor but not with the pT factor. Patients with central type lung cancer showed a significantly higher incidence of lymphatic recurrence than patients with the peripheral type. Patients having postoperative radiotherapy to prevent local recurrence showed a lower incidence of lymphatic recurrence than patients having no radiotherapy. In conclusion, lymphatic recurrence was the most frequent pattern in local recurrence after curative resection of lung cancer, and therefore improvements in the operative procedure of lymphatic dissection, as well as in postoperative adjuvant therapy including radiotherapy will be urgently required for the purpose of reducing local recurrence.

Adenocarcinoma↗

[Analysis of lymph node metastatic pattern in relation to prognosis and recurrence in pN2 resected lung cancer patients].

Lymph node metastasis was analyzed quantitatively with 4 categories and relation to post surgical survival and recurrence pattern was studied in patients with pN2 primary lung cancer who underwent relatively curative or relatively noncurative resection of the tumors. There was no relation between metastatic coefficient and post surgical survival, however, better survival was observed when the metastatic ratio and metastatic frequency were low and metastatic mode was random or skip pattern rather than sequential pattern. Metastatic coefficient and metastatic frequency were higher in cases with recurrence in lymph nodes but the former was lower and the latter was higher in cases with recurrence in intra-pulmonary dissemination or metastasis. There was no relation between metastatic coefficient and distant metastasis but metastatic frequency was lower in cases with recurrence in distant metastasis. Cases with sequential lymph node metastasis showed a tendency of lymph node recurrence and intrapulmonary metastasis and those with random or skip metastasis of lymph nodes had a tendency of distant metastasis.

Adenocarcinoma↗

Pituitary adenylate cyclase activating polypeptide (PACAP) with 27 residues. Conformation determined by 1H NMR and CD spectroscopies and distance geometry in 25% methanol solution.

The conformation of pituitary adenylate cyclase activating polypeptide with 27 residues (PACAP27) has been determined by two-dimensional NMR and CD spectroscopies and distance geometry in 25% methanol. Residues 9-20 and 22-25 have well-defined conformations but other residues do not show ordered conformations. The conformation of residues 9-20 is composed of three distinct regions of beta turn-like conformation (residues 9-12), alpha helix (residues 12-14) and the looser helical conformation (residues 15-20), while residues 22-24 form alpha helix. PACAP27 has a 2 helices separated by a disordered region similar to a VIP analog reported by Fry et al. but is distinct from the VIP analog in the position of the first helix, which is shifted by 2 residues toward the C-terminus, and in the form of the second helix [Fry, D.C., Madison, V.S., Bolin, D.R., Greeley, D.N., Toome, V. and Wegrzynski, B.B. (1989) Biochemistry 28, 2399-2409].

Amino Acid Sequence↗

[A quantitative approach to understand the ultramicro-structure responsible for input mechanism in excitation-contraction (E-C) coupling, on the basis of physio-morphological observations].

Our morphophysiological studies using concanavalin A-ferritin (Con A-F) have indicated that: (1) an out- and up-ward movement of a movable structure at the luminal surface-portion of the T-tubular membrane opposite the feet initiates contraction; (2) the grade of the movement depends on that of depolarization; (3) the movable structure is essentially a 'moving arm', which is fixed in wall of T-tubules at its fixed end and is able to be bound to the Con A-moiety of Con A-F particle about at its free end. Calculation based on molecular morphology and behaviour of Con A-F particle revealed following points: If (a) the origin of coordinate be the intersection of longitudinal center line of foot and the surface of T-tubular membrane in the transverse section of the tubules, (b) the fixed point of the arm is exactly on the surface of T-tubular membrane, and (c) the movement takes place in the transverse direction to the longitudinal axis of T-tubules, (1) the location of the center point of the movement of the moving arm is at 5.4 nm in the outside direction from the origin, (2) the arm is about 4 nm in length and moves by about 2.4 nm up- and out-ward at its free end upon about complete depolarization.

Animals↗

[A case of occult breast carcinoma with micronodular shadows on chest x-ray film].

A 57-year-old woman was admitted to our hospital for the evaluation of roentgenological micronodular shadows of the bilateral middle and lower lung fields. Physical findings were normal except for left axillary lymph adenopathy. The level of tumor marker NCC-ST-439 was markedly elevated (2800 U/ml). Histological examination of the transbronchial biopsy and the swollen lymph node biopsy specimens showed proliferation of identical tumor cells. Breast or lung carcinoma was suspected as the primary site. Echogram of the left breast showed microcalcification without a mass. The biopsy specimen obtained from the calcified lesion indicated occult breast carcinoma. From these findings the diffuse micronodular shadows on the chest roentgenogram were considered to be the metastatic lesions of occult breast carcinoma. This case indicates that occult breast carcinoma should be considered as a possible primary site for diffuse pulmonary metastatic lesions.

Antigens, Tumor-Associated, Carbohydrate↗

Epidermal growth factor (EGF) decreased endothelin-2 (ET-2) production in human renal adenocarcinoma cells.

Production of immunoreactive (ir-) endothelin-2 (ET-2) in renal adenocarcinoma cells, ACHN, was reduced by transforming growth factor-beta, basic fibroblast growth factor, transforming growth factor-alpha and, most strikingly, by epidermal growth factor (EGF). These growth factors did not show such inhibitory effects on the secretion of ir-ET-1 in ET-1-producing cells, indicating that the production of ET-2 and ET-1 is regulated differently by the growth factors. EGF specifically reduced the secretion of not only ir-ET-2 but also ir-big ET-2 with only a small decrease in total protein synthesis. Northern blot analysis indicated that EGF controls the ET-2-production at the transcription levels of ET-2 gene.

Adenocarcinoma↗

Autoradiographic distribution of pituitary adenylate cyclase activating polypeptide (PACAP) binding sites in the rat brain.

Distribution of pituitary adenylate cyclase activating polypeptide (PACAP) binding sites was investigated in the rat brain and pituitary gland by means of in vitro autoradiography. High densities of specific [125I]PACAP binding were observed in the anterior pituitary, hippocampus (CA1-4 and dentate gyrus) and in the superior colliculus. Moderate to high labeling was observed in the periaqueductal gray matter, substantia nigra pars compacta, and in the habenula. The hypothalamus, thalamus, ventral tegmental area (VTA), mammillary body and medial geniculate body were moderately labeled. The present results support possible actions of PACAP on the pituitary functions, and further suggest that PACAP is a neurotransmitter/neuromodulator in the central nervous system.

Animals↗

Chemotactic peptide from ropalidian wasp as well as the authentic chemotactic tripeptide stimulates two distinct pathways in neutrophils, but the [LYS7] analog does only one of them.

In our previous study on chemotactic peptide isolated from Ropalidian wasp, it was found to induce not only chemotaxis but also other cellular responses, such as superoxide generation and lysosomal enzyme release, but [Lys7] analog was found to induce only chemotaxis but not others. Here, studies on intracellular Ca2+ changes and receptor-binding revealed that the wasp chemotactic peptide and the authentic tripeptide stimulated two distinct receptors, but that the [Lys7] peptide could have access to only one of them.

Animals↗

Assignment of disulphide bonds in synthetic endothelin-1 isomers by fast atom bombardment mass spectrometry.

Endothelin-1 (ET-1), a 21-residue vasoconstrictor peptide possessing four cysteinyl residues at positions 1, 3, 11 and 15, was synthesized by random oxidation of a tetrahydro-ET-1. On reverse-phase high-performance liquid chromatography, crude product was shown to be a mixture of two disulphide isomers. A method was developed to determine the disulphide structure of the isomers. The method consisted of (a) limited digestion with chymotrypsin, (b) cleavage with cyanogen bromide and (c) manual Edman degradation. Through this procedure, each isomer afforded specific fragments containing a single disulphide bond, which were identified by fast atom bombardment mass spectrometry. Isomer 1, the minor component, afforded a fragment containing Cys 3 and Cys 15, and isomer 2, the major component, afforded fragments containing Cys 3 and Cys 11. Since little disulphide exchange was observed, it could be concluded clearly that the disulphide bond pairs in isomer 1 were Cys 1-Cys 11 and Cys 3-Cys 15, while those in isomer 2 were Cys 1-Cys 15 and Cys 3-Cys 11 (the same as natural ET-1). The procedure was successfully applied to two synthetic analogues, [Gly18]-ET-1 and [Pro16]-ET-1.

Disulfides↗

Comparison of endothelin-1 concentrations in normal and complicated pregnancies.

Endothelin-1 (ET-1) may be important in regulating vascular tone in humans. To understand the role of ET-1 during pregnancy, we measured the concentrations of plasma immunoreactive ET-1 in normal, toxemic (pre-eclamptic or eclamptic), and other complicated pregnancies. There were no significant differences between ET-1 concentrations in normal pregnancy and those in toxemic pregnancy, except in one case with IgA nephropathy and severe toxemia. There also was no significant difference between the ET-1 concentrations in pure toxemia of pregnancy compared with superimposed toxemia of pregnancy. Furthermore, there was no correlation between ET-1 concentrations, blood pressure, or renal function during toxemic pregnancy. Similarly, there were no significant differences between ET-1 plasma levels in normal pregnancies and other complicated pregnancies except for two cases in which much blood was lost due to uterine atony and abruptio placenta. Thus, plasma ET-1 concentrations showed no significant relationship to blood pressure or renal function during normal or toxemic pregnancy, and were not related to the type (pure or superimposed type), grade, or severity of toxemic pregnancy.

Adult↗

Increased plasma concentrations of endothelin-1 during and after pulmonary surgery.

We studied the involvement of endothelin-1 (ET-1) in the physiological response to surgical stress. Plasma concentrations of ET-1 were measured by a sandwich-type enzyme immunoassay. The blood samples were collected from the pulmonary artery (PA), the left atrium (LA), and the median cubital vein (MCV) in four patients who had undergone pulmonary lobectomy. The samples were collected before, during, and after surgery. Plasma ET-1 concentrations increased in the blood from all sampling sites. The highest concentration of plasma ET-1 was observed at 6 h after surgery in MCV (3.97 +/- 1.47 pg/ml, mean +/- SD) and at the end of surgery in PA (1.79 +/- 0.48 pg/ml) and in LA (1.93 +/- 0.43 pg/ml). These values returned to the baseline value within 72 h after surgery [1.21 +/- 0.19 (MCV), 1.00 +/- 0.18 (PA)]. Although the lung has a large capacity to remove ET-1 from the circulating blood, ET-1 concentrations measured in plasma samples obtained from the LA were not different from those obtained from the PA. This may suggest that the lungs have the ability to absorb and release ET-1 simultaneously during pulmonary surgery. The increase of ET-1 in MCV was greater than that in PA or in LA. This suggests the possibility of ET-1 release from the forearm.

Endothelins↗

Sandwich-enzyme immunoassays for endothelin family peptides.

Sandwich-enzyme immunoassays (sandwich-EIAs) for endothelin-1 (ET-1), endothelin-3 (ET-3), big ET-1, big endothelin-2 (big ET-2), and big endothelin-3 (big ET-3) have been established using two antibodies directed against the N-terminal and C-terminal portions of each ET and big ET. These sandwich-EIAs are sensitive enough to detect 0.1-0.4 pg/well of each ET or big ET with cross-reactivity of less than 1% with other ETs and big ETs (except in the case of the EIA for ET-1, which detects immunoreactivity of ET-2 as well as ET-1). Plasma levels of immunoreactive (ir-) ET-1, big ET-1, and ET-3 in normal humans were approximately 2.0, 5.1, and 0.9 pg/ml, respectively, when corrected for recovery efficiency. In reverse-phase high-performance liquid chromatography (HPLC) analysis, immunoreactivities detected by these sandwich-EIAs were eluted at the positions of authentic ET-1, big ET-1, and ET-3, indicating that plasma levels of ET-1, big ET-1, and ET-3 were precisely measured by our methods. Levels of ir-big ET-2 and ir-big ET-3 were also detected in normal human plasma; however, further studies are needed to determine their precise values. The establishment of reliable sandwich-EIAs and the measurement of specific ETs and big ETs in normal human plasma will contribute to clarifying the biological and clinical significance of ET family peptides.

Adult↗

Contribution of endogenous endothelin to the extension of myocardial infarct size in rats.

Pathophysiological roles of endogenous endothelin have been studied from the viewpoint of its contribution to the extension of myocardial infarct size. A monoclonal antibody against endothelin 1 (AwETN40) suppressed changes induced by endothelin 1 and endothelin 2 but did not modify those by endothelin 3 in vivo or in vitro. Effects of AwETN40 on myocardial infarct size were investigated. Coronary ligation (1 hour) and reperfusion (24 hours) in rats caused infarction in 35% of the left ventricle. Repetitive or single administration of AwETN40 reduced the infarct size; an intravenous injection of 22.5 mg/kg of the antibody 5 minutes after coronary occlusion or 5 minutes before reperfusion reduced the size by 38% or 31% of the control, respectively. Plasma and tissue endothelin 1 and plasma big endothelin 1 in rats were measured at various stages after occlusion. Plasma endothelin 1 showed a fourfold increase 10 minutes after reperfusion (from 1.02 to 3.96 pg/ml) and had returned to the control value after 8 hours. Plasma big endothelin 1 showed changes similar to those of plasma endothelin 1. No significant changes in plasma endothelin 2 and endothelin 3 were observed. Cardiac tissue contained seven times as much endothelin 1 as the control value 1 hour after reperfusion (4.59 versus 33.1 pg/g tissue), and a high concentration (13.2 pg/g tissue) was maintained even after 48 hours. We concluded that an increase in endogenous endothelin 1 plays an important role in the extension of myocardial infarct size.

Animals↗

[Clinical backgrounds of six cases of childhood tuberculosis].

Clinical backgrounds of six children who entered Matsue National chest Hospital due to pulmonary tuberculosis and tuberculous pleuritis were investigated and the following results were obtained. 1) Of six children five had not received BCG vaccination. 2) There were two children who developed tuberculosis after the completion of chemoprophylaxis. 3) None was detected by the examination of the household of the source cases. 4) Both patients and doctors were responsible for the delay in detecting the source, which ranged between five and seventeen months in total. 5) Among the source cases there were two patients who were smear-negative but culture-positive. In view of the above observations it is suggested that BCG vaccination is important for the prevention of tuberculosis in children and that follow-up examinations should be continued after the completion of chemoprophylaxis. It should also be emphasized that early detection of the source case is imperative for the prevention of tuberculosis.

BCG Vaccine↗