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Biomedical subjects

M Takagi

Publications and source records attributed to M Takagi.

At least 721 records · Page 40Linked to original sources

Time course analyses of kinins and other mediators in plasma exudation of rat kaolin-induced pleurisy.

Pleurisy was induced in rats by an intrapleural injection of 0.5 ml of 1% kaolin. The exudation of plasma into the pleural cavity showed two peaks at 20 min and 3-5 h after the kaolin injection. The volume of the pleural fluid increased gradually up to 5 h. The effects of treatment with mepyramine, methysergide, captopril, bromelain and indomethacin suggested that the early phase (20 min) of exudation was mediated mainly by kinins, histamine and 5-HT, and that the late phase (3 h) was mediated by prostaglandins (PGs) and possibly kinins. We measured the levels of histamine, kinin and PG in the pleural exudate to verify the involvement of the mediators mentioned above. Intracellular histamine levels decreased markedly and extracellular histamine levels increased significantly 20 min after the induction of kaolin pleurisy. Only threshold levels of kinin were detected after the induction of pleurisy. Captopril treatment, however, increased kinin levels which peaked at 20 min and decreased rapidly thereafter. Levels of 6-keto-PGF1 alpha and thromboxane B2 showed a peak at 20 min, whereas levels of PGE2 increased gradually from 20 min to 5 h. These results indicate that kaolin-induced pleurisy is a kinin-related inflammation and could be used as a model for studying the in vivo interaction of the kallikrein-kinin system and PGs at inflammatory sites.

Animals↗

Ultrastructural cytochemistry of trout arterial fibrils as elastic components.

The trout arterial wall contains numerous extracellular fibrils that are presumed to be elastic. However, the cytochemical properties of the arterial fibrils have not been studied. Thus, we have ultrastructurally and cytochemically examined these fibrils, utilizing routine uranyl acetate and lead (UA-Pb) double staining, the tannic acid (pH 7.0)-uranyl acetate (TA-UA) method as an electron-dense staining for elastin, and Thiéry's periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method to localize vicinal glycol-containing complex carbohydrates. The arterial fibrils, about 23 nm in thickness, were interwoven at random but frequently showed the circular alignment to the long axis of the aorta. Occasionally they appeared to coalesce side by side, and the coalesced portion tended to lose its affinity for UA-Pb stains. The TA-UA method stained the fibrils moderately to intensely and stained the coalesced parts of the fibrils more intensely. All of those TA-UA positive fibrils were completely removed after elastase en bloc digestion. The PA-TCH-SP method did not stain the arterial fibrils but stained another kind of much thinner interfibrillar filamentous structure. These results suggest that the fibrils in the wall of trout ventral aorta are elastin in nature and do not contain vicinal glycols, although the fibrils usually exist in a fibrillar form, which is unlike mammalian amorphous elastin.

Animals↗

Strength of bonding between A-W glass-ceramic and the surface of bone cortex.

We examined the bonding strength of apatite- and wollastonite-containing glass-ceramics (A-W.GC) to the surface of bone cortex. Semi-column-shaped blocks of A-W.GC were fixed with screws on the surface of the tibiae of rabbits. The blocks of alumina-ceramic were used as the control. The load required to detach the implant from the surface of bone cortex was measured 2, 4, 8, and 25 weeks after the implantation. The bonding strength between A-W.GC and the surface of bone cortex increased with time. It increased remarkably 4 weeks after the implantation, suggesting that the chemical bonding advanced rapidly between 2 and 4 weeks after the implantation. The tensile strength of bonding reached a maximum of 15.1 +/- 3.1 kg/cm2 25 weeks after the implantation. The shear strength of bonding reached a maximum of 17.1 +/- 2.3 kg/cm2 25 weeks after the implantation. From these studies, we conclude that A-W.GC has a capability of making relatively quick bonding to the surface of bone cortex.

Animals↗

Construction of promoter-probe vectors for Candida maltosa, a n-alkane-assimilating yeast, using the LEU2 gene of Saccharomyces cerevisiae.

For the purpose of isolation of promoter regions which are regulated by a carbon source in the medium in an n-alkane-assimilating yeast, Candida maltosa, two promoter-probe vectors were constructed. Each of them consists of the LEU2 gene of Saccharomyces cerevisiae whose 5'-noncoding region was trimmed with BAL31, an autonomously replicating sequence isolated from C. maltosa genome (the TRA region) which we have previously isolated, and the pBR322 sequence. One of them, pPLC2, having the TATA box, lacks the regulatory sequence ("sequence L") of the LEU2 gene, and the other, pPLC1, lacks both the TATA box and sequence L. Using pPLC1 as a short-gun cloning vector in C. maltosa, many promoter regions which were active when glucose was present in the medium as a carbon source were obtained from the genome of C. maltosa. The sizes of the inserted fragments of two of them were determined. (In this paper, a promoter region refers to a promoter which includes a TATA box, plus a regulatory sequence such as an UAS (upstream activating sequence)-like sequence).

Candida↗

Specific binding of peanut agglutinin and soybean agglutinin to chondroitinase ABC-digested cartilage proteoglycans: histochemical, ultrastructural cytochemical, and biochemical characterization.

The binding of peanut agglutinin (PNA) and soybean agglutinin (SBA) to cartilage proteoglycans was investigated by histochemical, ultrastructural cytochemical, and biochemical methods. Following aldehyde fixation, specimens of rat epiphyseal cartilage were examined by horseradish peroxidase-labelled lectin cytochemistry with and without prior digestion in chondroitinase ABC. At the light microscope level neither PNA nor SBA exhibited any affinity for cartilage matrix, but became strongly bound following chondroitinase treatment. Similarly, at the ultrastructural level, extracellular matrix granules, presumed to be proteoglycan monomer(s), lacked PNA affinity in undigested specimens, and stained very weakly with SBA. Both PNA and SBA weakly to moderately stained the trans cisternae of the Golgi-flattened cisternae in chondrocytes. The chondrocyte plasmalemma lacked PNA staining, but reacted weakly with SBA. Following chondroitinase digestion, PNA and SBA stained matrix granules, and the cell surface of chondrocytes intensely, whereas the Golgi trans cisternae, the Golgi-derived vacuoles, and multivesicular bodies demonstrated weak to moderate reactivity. Proteoglycan aggregates purified from rat chondrosarcoma and bovine nasal cartilage bound PNA and SBA avidly after digestion with chondroitinase. Undigested proteoglycans lacked affinity for PNA and reacted very weakly with SBA. These results indicate that both PNA and SBA specifically react with chondroitinase-modified oligosaccharide(s) bound to core proteins of cartilage proteoglycans. This provided a specific histochemical and ultrastructural cytochemical procedure for localizing chondroitin sulphate-containing proteoglycans.

Animals↗

Development of inactivated vaccine against virus causing haemorrhagic fever with renal syndrome.

B-1 virus belonging to the hantavirus group was serially passaged in the brains of newborn mice. Inactivated vaccine was prepared from the brains after inactivation with formalin and then purification by ultracentrifugation. The antigenic potency of this vaccine in vitro was determined by antibody-bound enzyme-linked immunosorbent assay (ELISA) and serial diluted vaccine bound to an aluminium hydroxide gel was inoculated into Balb/c mice to test immunogenicity. After two injections of this vaccine preparation, antibodies were detected in the mice by immunofluorescent, neutralizing and haemagglutination inhibition antibody tests. When mice immunized with this vaccine were challenged with B-1 virus and Hantaan virus (KHF-83-61BL strain), the virus titres in their lungs and spleens were significantly less than those in non-immunized mice. These results suggest that inactivated B-1 virus vaccine is effective against virus challenge by homotypic (B-1 virus) and heterotypic (Hantaan virus) viruses.

Animals↗

Primary structure of a base non-specific ribonuclease from Rhizopus niveus.

The primary structure of a base non-specific ribonuclease from Rhizopus niveus (RNase Rh) was determined by nucleotide sequence analysis of the DNA fragment encoding RNase Rh gene including signal peptide sequence, and amino acid sequence analysis of the peptide obtained from RNase Rh and RNase Rh' (a protease-modified RNase Rh created during the course of purification). The sequence determined was: MKAVLALATLIGSTLASSCSSTA LSCSNSANSDTCCSPEYGLVVLNMQWAPGYGPANAFTLHGLWPDKCSGAYAPSGGCDSN RASSSIASVIKSKDSSLYNSMLTYWPSNQGNNNVFWSHEWSKHGTCVSTYDPDCYDNYE EGEDIVDYFQKAMDLRSQYNVYKAFSSNGITPGGTYTATEMQSAIESYFGAKAKIDCSSG TLSDVALYFYVRGRDTYVITDALSTGSCSGDVEYPTK (the sequence of signal peptide is underlined). The sequence indicates that the homology with the sequence of RNase T2 from A. oryzae with the same base specificity is about 42% and that the sequences around the two histidine residues which are supposed to be involved in the active site are fairly conserved.

Amino Acid Sequence↗

Dopamine and aldosterone in adrenal glomerulosa cells in spontaneously hypertensive rats.

To investigate the possible significance of dopamine in aldosterone production, the adrenal zona glomerulosa was examined for the presence of dopamine and aldosterone in spontaneously hypertensive rats (SHR) and Wistar-Kyoto rats (WKY) aged 10 weeks. Aldosterone and dopamine were measured in the adrenal capsular portions (mostly zona glomerulosa) by radio-immunoassay and high-performance liquid chromatography, respectively. Dopamine was found in the adrenal zona glomerulosa, with a significantly higher level in SHR than in WKY, whereas the content of aldosterone was significantly lower in SHR than in WKY. Furthermore, there was a significant negative relationship between the content of dopamine and that of aldosterone. These results suggest that the inhibitory effect of dopamine on aldosterone production may be mediated within the adrenal zona glomerulosa and that an increased dopamine content in the adrenal zona glomerulosa may contribute to the decreased aldosterone content in SHR.

Adrenal Glands↗

Isolation and sequencing of a genomic clone encoding aspartic proteinase of Rhizopus niveus.

A gene encoding Rhizopus niveus aspartic proteinase was isolated from an R. niveus genomic library by using oligonucleotides probes corresponding to its partial amino acid sequence, and its nucleotide sequence was determined. By comparing its deduced amino acid sequence with the amino acid sequence of rhizopuspepsin (5, 26), we concluded that the R. niveus aspartic proteinase gene has an intron within its coding region and that it has a preproenzyme sequence of 66 amino acids upstream of the mature enzyme of 323 amino acids.

Amino Acid Sequence↗

Effect of atrial natriuretic factor on calcium fluxes in adrenal glomerulosa cells.

We studied the effect of atrial natriuretic factor (ANF) on calcium influx and efflux in rat adrenal glomerulosa cells stimulated by angiotensin II (Ang II) or potassium ion, and observed how ANF inhibits the initial and sustained phases of the aldosterone response to Ang II or K+ using a superfusion system of dispersed adrenal glomerulosa cells. K+ (8 mM) significantly increased Ca2+ influx rate compared with basal rate (0.91 +/- 0.10 vs 0.42 +/- 0.04 nmol/min/10(6) cells; p less than 0.01). ANF (10(-8) M) did not inhibit the K+-induced increase in Ca2+ influx rate (0.99 +/- 0.18 nmol/min/10(6) cells). Ang II (10(-9) and 10(-8) M) stimulated Ca2+ influx rate (10(-9) M Ang II, 0.62 +/- 0.02; 10(-8) M Ang II, 0.71 +/- 0.09 vs basal, 0.44 +/- 0.03 nmol/min/10(6) cells; p less than 0.05), while ANF (10(-8) M) did not change the Ca2+ influx rate increased by Ang II (ANF + 10(-9) M Ang II, 0.62 +/- 0.06; ANF + 10(-8) M Ang II, 0.69 +/- 0.14 nmol/min/10(6) cells). In the Ca2+ efflux study ANF (10(-8) M) was perfused through the cells 10 minutes before the start of perfusion with Ang II (10(-9) M) or K+ (12 mM).(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Glands↗

Ultrastructural visualization of selective peanut agglutinin binding sites in rat osteoclasts.

We investigated the distribution of concanavalin A (ConA)-reactive alpha-D-mannosyl and alpha-D-glucosyl groups and peanut agglutinin (PNA)-reactive beta-D-galactose-(1----3)-N-acetyl-D-galactosamine residues on the surface of osteoclasts with pre-embedment ultrastructural lectin cytochemistry after aldehyde fixation of the metaphyses of the rat tibiae. By routine morphology, the plasma membrane of the ruffled border of the osteoclast was distinguished from the rest of the cell membrane, with the exception of the membrane of coated pits, by its characteristic thick coat at its cytoplasmic surface. Cytochemistry, using ConA in combination with horseradish peroxidase (ConA-HRP) and PNA conjugated to HRP, showed that binding of ConA was distributed over the entire cell surface of osteoclasts. In contrast, intense binding of PNA was limited to the membranes of the ruffled border and coated pits, whereas the remainder of the cell membrane stained weakly or not at all. These results demonstrate that preferential PNA binding sites of the cell surface correspond to coated membranes associated with osteoclastic endocytosis.

Animals↗

Effect of atrial natriuretic peptide on renin release in a superfusion system of kidney slices and dispersed juxtaglomerular cells.

The effect of atrial natriuretic peptide (ANP) on renin release is controversial. Several reports state that ANP inhibits renin secretion, while others have shown no effect. We investigated the effect of synthetic rat ANP with 24 amino acids (atriopeptin III) on renin release in vitro in a dynamic superfusion system of renal cortical slices as well as collagenase-dispersed juxtaglomerular cells. In the superfusion system of kidney slices, isoproterenol (5 x 10(-8) M) clearly stimulated renin release from kidney slices, while angiotensin II (AII; 10(-5) M) suppressed renin release. ANP (10(-10)-10(-6) M) did not inhibit basal renin release or blunt the stimulatory effect of isoproterenol. The suppression of renin secretion by AII was never modified in the presence of ANP. The superfusion system of juxtaglomerular cells demonstrated greater sensitivity of renin release in responses to isoproterenol and AII. In this system, ANP (10(-6) M) did not alter renin release from the cells stimulated by isoproterenol (5 x 10(-8) M) or inhibited by AII (10(-8) M). However, basal renin release was slightly stimulated in the late phase of ANP superfusion and for 20 min after the ANP perfusion was stopped. Similarly, 8 bromo-cGMP (10(-6) M) did not inhibit, but, rather, stimulated basal renin release slightly. These results suggest that ANP does not inhibit renin release by a direct effect on the juxtaglomerular cell in the rat.

Angiotensin II↗

Effects of dibutyryl adenosine 3',5'-monophosphate, angiotensin II, and atrial natriuretic factor on phosphorylation of a 17,600-dalton protein in adrenal glomerulosa cells.

Rat adrenal glomerulosa cells were incubated with [32P]phosphate and (Bu)2AMP (dbcAMP), angiotensin II, and atrial natriuretic factor (ANF). Incorporation of [32P]phosphate into cellular proteins was analyzed by one-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. dbAMP stimulated phosphorylation of a 17.6K protein, while angiotensin II did not stimulate it. ANF did not affect the protein phosphorylation, whether the cells were in the basal state or stimulated by dbcAMP or angiotensin II. On the other hand, ANF markedly inhibited angiotensin II-stimulated aldosterone production, but only slightly inhibited dbcAMP-stimulated aldosterone. These results suggest that in rat adrenal glomerulosa cells phosphorylation of the 17.6K protein may have a relationship with the stimulatory effect of cAMP on aldosterone production; however, neither angiotensin II nor ANF affected the phosphorylation of this protein, and phosphorylation of the 17.6K protein is not an obligatory step in the regulation of aldosterone production.

Aldosterone↗

Effects of intracerebroventricular infusion of aldosterone on blood pressure and sodium and potassium concentrations in cerebral spinal fluid in rats.

The central effects of aldosterone on blood pressure and sodium (Na) and potassium (K) concentrations in cerebral spinal fluid (CSF) were examined by chronic intracerebroventricular (ICV) infusion of aldosterone. The rats were infused for 14 days with either ICV artificial CSF, ICV aldosterone or aldosterone subcutaneously using miniosmotic pumps. ICV aldosterone increased blood pressure and lowered the K concentration in CSF. The Na concentration in CSF tended to increase, although not significantly. No significant changes in concentrations of serum Na and K, plasma renin, aldosterone, norepinephrine or ANP were observed. These results suggest that aldosterone induces blood pressure elevation through a central action, and that a change in Na or K concentration in CSF may be involved.

Aldosterone↗

Effects of active oxygen scavengers on the peroxidation of linoleic acid catalyzed by dehydro-L-ascorbic acid or its degradation products.

The addition of 1,4-diazabicyclo-[2,2,2]octane (DABCO) (100 mM) or 1,2-dihydroxybenzene-3,5-disulfonic acid (Tiron) (1 mM) to a reaction mixture containing 10 mM linoleic acid (LA), 20% EtOH, and 135 microM dehydroascorbic acid (DHA) as a catalyst suppressed LA peroxidation, but the addition of mannitol (approximately 100 mM), uric acid (100 microM), and catalase (6.5 units) did not. DHA or 2,3-diketo-L-gulonic acid (DKG) accelerated LA peroxidation, but the splitting products of DHA did not affect LA peroxidation. These results suggest that some specific radicals are liberated in the degradation of DHA or DKG.

1,2-Dihydroxybenzene-3,5-Disulfonic Acid Disodium ↗

SF2446, new benzo[a]naphthacene quinone antibiotics. I. Taxonomy and fermentation of the producing strain, isolation and characterization of antibiotics.

New antibiotics SF2446A1, A2, A3, B1 and B2 have been isolated from the culture of Streptomyces sp. SF2446 and antibiotic SF2446B3 has been obtained by methanolysis of SF2446B1 or B2. SF2446A1, A2 and B1 showed strong inhibitory activities against mycoplasmas and Gram-positive bacteria. Empirical molecular formulae of antibiotics SF2446-A1, A2, A3, B1, B2 and B3 were determined to be C34H35NO15, C26H21NO11, C34H35NO14, C34H35NO14 and C26H21NO10, respectively.

Aminoglycosides↗