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Biomedical subjects

Y Toda

Publications and source records attributed to Y Toda.

At least 19 recordsLinked to original sources

Immunoelectron microscopic studies of glycosaminoglycans in the metaphyseal bone trabeculae of growing rats.

The types and distribution of glycosaminoglycans (GAGs) were studied immunocytochemically in osteoid, mineralized bone matrix, and cartilage matrix of growing rat metaphyseal bone after aldehyde fixation and EDTA demineralization, using four monoclonal antibodies (mAbs 1-B-5, 2-B-6, 3-B-3 and 5-D-4). These mAbs specifically recognize epitopes in non-sulphated chondroitin (C0-S); chondroitin 4-sulphate (C4-S) and dermatan sulphate (DS); chondroitin 6-sulphate (C6-S) and C0-S; and keratan sulphate (KS) respectively. In osteoid, all mAbs except 1-B-5 weakly stained matrix material on and between collagen fibrils, and moderately stained organic material corresponding to bone nodules, which are known sites of mineralization. However, the staining of osteoid abruptly decreased at the mineralization front; weak staining was confined mostly to the organic material of bone nodules in mineralized bone matrix, with very weak or no staining of the rest of the bone matrix. This staining progressively decreased toward the mineralized cartilage matrix and became negative. The mineralized cartilage matrix and lamina limitans reacted strongly with all mAbs except 5-D-4. These results indicate that osteoid contains sulphated proteoglycans containing C4-S and/or DS, C6-S and KS, and subsequent bone matrix mineralization appears to require accumulation of these macromolecules within bone nodules and eventual loss of these substances for complete mineralization, whereas proteoglycans containing C0-S, C4-S and/or DS, and C6-S still exist in mineralized cartilage matrix and lamina limitans.

Animals

Structural determination of gangliosides that bind to influenza A, B, and C viruses by an improved binding assay: strain-specific receptor epitopes in sialo-sugar chains.

An improved binding assay for detection of ganglioside receptors for influenza A, B, and C viruses was developed. In this system, the virions bound to gangliosides that were developed on a silica gel thin-layer plate were detected by mouse monoclonal antibody against viral hemagglutinin and peroxidase-conjugated anti-mouse immunoglobin. No hydrolysis of the gangliosides by viral receptor-destroying enzyme was detected in the present condition. The reactivity of the viruses to gangliosides depended on the amount of developed gangliosides (10 pmols-10 nmols), the molecular species of sialic acid, and their sugar sequences. Human influenza A (PR/8/34), B (Lee/40), and C (Ann Arbor/1/50) viruses bound different receptor epitopes of sialo-sugar chains of gangliosides. The A/PR/8 virus bound most effectively to Neu5Ac-containing lacto-series gangliosides carrying type I and type II sugar chains, followed by ganglio-series and hematoside-series gangliosides. The A/PR/8 virus weakly bound to Neu5Ac alpha 2,6lactotetraosylceramide [IV6(Neu5Ac)Lc4Cer] and Neu5Ac alpha 2,6paragloboside [IV6(Neu5Ac)nLc4Cer] carrying Neu5Ac alpha 2,6Gal sequence, although their Neu5Ac alpha 2,3Gal derivatives were the most potent gangliosides tested. B/Lee/40 bound restrictively to IV6(Neu5Ac)Lc4Cer and IV6(Neu5Ac)nLc4Cer, which carry Neu5Ac alpha 2,6Gal sequence, and type I and type II lacto-series sugar chain, respectively. C/Ann Arbor/1/50 reacted only with 9-O-Ac-Neu5Ac-carrying sugar chains in all the gangliosides tested. This method also allowed the microanalysis of receptor gangliosides of unknown samples. ESK cells, sensitive to the influenza A viruses infection, expressed several kinds of receptor active gangliosides, while those from ESK-R cells, resistant to the virus infection, were undetectable.

Binding Sites

Siblings of 21-hydroxylase deficiency (non-salt-losing) with aldosterone hypersecretion.

We describe siblings with the non-salt-losing form of 21-hydroxylase deficiency who had hypersecretion of aldosterone and plasma renin activity (PRA). Blood pressure and serum electrolytes in both cases were normal despite the aldosterone hypersecretion. Aldosterone secretion was elevated markedly with ACTH administration and with sodium deprivation and/or volume depletion during ACTH suppression by dexamethasone. With suppression by dexamethasone, aldosterone hypersecretion was decreased with lowering of the steroids proximal to the block in the biosynthetic pathway. However, urinary sodium excretion was decreased. These results suggest that the biosynthetic pathway for aldosterone production was preserved. Furthermore, aldosterone hypersecretion and high PRA may serve to compensate for the sodium loss which results in turn from the overproduction of the sodium-losing steroids, such as progesterone and 17 alpha-hydroxyprogesterone which are aldosterone antagonists.

Adolescent

Studies on viridenomycin, a novel 24-membered macrocyclic polyene lactam antibiotic.

A new antitumor antibiotic, designated AL081, was obtained from the culture filtrate of an actinomycete identified as Streptomyces gannmycicus, and found to be identical with viridenomycin by direct comparison. The structure of the antibiotic was determined by NMR spectral analysis including a variety of two-dimensional techniques to be a novel 24-membered macrocyclic polyene lactam. Viridenomycin prolonged the survival periods of mice bearing P388 leukemia and B16 melanoma cells.

Animals

[The effect of anesthetic agents on descending spinal cord evoked potential and the compound muscle action potentials elicited by stimulation at the cerebral motor cortex and the spinal cord].

Recently, intraoperative monitoring of the motor tract by descending spinal cord motor evoked potentials (MEP) and compound muscle action potentials (CMAP) has been applied in clinical testing. Since several reports have mentioned the vulnerability of these potentials to anesthetic agents, experimental studies were carried out on the relationship between these potentials and anesthesia using 41 adult cats. The effects of anesthesia on changes in amplitude of the direct wave (D wave) and indirect wave (I wave) of the MEP and CMAPs were investigated. These potentials were generated by stimulation of the spinal cord and the motor cortex, respectively. Enflurane (2%), halothane (1%) and isoflurane (1.5%) with pure oxygen decreased the amplitude of the I wave to less than 50% of the control level. The CMAP after stimulation of the spinal cord was degraded to less than 30%, and the CMAP after cortical stimulation vanished completely. Only the D wave was stable against inhalational anesthetic agents. Sixty-seven percent nitrous oxide with the above concentrations of these inhalational anesthetic agents decreased the amplitude of the I wave to less than 30% and the CMAP evoked by spinal cord stimulation vanished. The effect of modified NLA (diazepam and pentazocine) on these potentials was weaker than that of the inhalational anesthetic agents.

Action Potentials

Impairment of bone formation with aluminum and ferric nitrilotriacetate complexes.

The deleterious effects of aluminum(AL) and iron(Fe) on bone formation were studied in the presence of nitrilotriacetate (NTA) as a chelator. Both Al-NTA (1.0-1.5 mg Al/kg/day, n = 12)- and ferric nitrilotriacetate (Fe-NTA) (2.0 mg/kg/day, n = 4)-treated Wistar rats showed renal insufficiency blood urea nitrogen [BUN] levels of 25 +/- 8.8-20 +/- 0.7 compared to 12 +/- 0.7-11 +/- 0.4 mg/dl), osteomalacia with a relative osteoid volume of 31.5 +/- 5.6-13.2 +/- 2.4 compared to 4.6 +/- 1.8-0.83 +/- 0.12%, and bone growth retardation (3.1 +/- 0-3.0 +/- 0.2 compared to 3.4 +/- 0-3.3 +/- 0.1 cm) in 24 control rats. Dietary vitamin E(VE) supplementation prevented the Fe-NTA-induced impairment, but not the Al-NTA toxicity. Aluminum was deposited at the interface between osteoid and mineralized bone, while Fe was deposited in the osteoblasts and osteoclasts. There seems to be a positive correlation between hypophosphatemia and osteomalacia but carboxy-terminal parathyroid hormone (C-PTH) and calcium (Ca) levels in the serum were not related to the degree of osteomalacia. Administration of Al-NTA results in more bone Al deposition than that of aluminum chloride (AlCl3) (450 +/- 40 compared to 211 +/- 18 mg/kg fat-free dry weight). The Fe-NTA bone change is related to VE-preventable cellular injury, being consistent with the notion that Fe-NTA toxicity is caused by lipid peroxidation. Al-NTA can be used as an animal model of renal osteodystrophy. Osteodystrophy by Al in chronic renal failure may be mediated by the intrinsic chelator or chelating substance(s) retained in the body fluid due to renal insufficiency.

Aluminum

Distribution and immunochemical specificities of fimbriae of Porphyromonas gingivalis and related bacterial species.

Rabbit polyclonal antibody (Poly-1) and mouse monoclonal antibodies (mAbs) TO-11, TO-14 and TO-M1 specific for Porphyromonas gingivalis 381 fimbriae were prepared. Poly-1 and the 3 mAbs were screened for their reactivity with whole cells oral and nonoral black-pigmented bacterial species by enzyme-linked immunosorbent assay (ELISA) and the binding experiment using [125I]Poly-1 and [125I]mAbs. ELISA revealed that Poly-1 definitely reacted with whole cells of all the 11 strains of P. gingivalis tested. However, 8 of 11 P. gingivalis strains reacted with mAbs TO-11, TO-14 and TO-M1. These results were confirmed by the specific binding of radiolabelled Poly-1 and mAb TO-11 to the 8 strains. The M(r) of the fimbrial subunit protein (fimbrilin) isolated and purified from P. gingivalis strains 381, BH18/10, HW24D-1, 6/26 and OMZ 314 was 41 kDa by sodium dodecylsulfate-polyacrylamide gel electrophoresis. It was found by immunoblotting that mAbs TO-11 and TO-14/TO-M1 recognized different epitopes of fimbrial protein from P. gingivalis strains. Immunoelectron micrographs of whole cells and the purified fimbriae of P. gingivalis strains visualized similar serotype-specific antibody bindings to the fimbriae. These results indicate that 11 strains of P. gingivalis could be divided into at least 2 separate groups based on the immunochemical specificities of the fimbriae.

Animals

Effects of color CRT display on pupil size in color-blind subjects.

The effects of color cathode ray tube (CRT) display on the pupil size in color-blind subjects were studied experimentally. Red, magenta, green, cyan, yellow, and white were presented on the CRT. Five protan subjects, five deutans, and five normal subjects were tested using infrared pupillography. In the protan group, the pupils were significantly less sensitive to red (wavelength: 600 nm) targets compared to the other colors. In the deutans and the control group, there were non-significant changes in pupil size in response to the colors. Dominant wavelengths above 600 nm in color CRTs should be avoided in routine work presentation because of less sensitivity in protanopias.

Adult

[Study of prognostic factors of survival in patients with unresectable non-small cell lung cancer].

We studied prognostic factors of survival in 121 patients unresectable non-small cell lung cancer treated between March 1983 and November 1988 at forth department of internal medicine, Kinki university school of medicine, about histology, clinical stage, age, sex, performance status, Brinkman Index, hemoglobin, serum TP, serum Alb, serum LDH, pulmonary function and chemotherapy. The prognostic factors were studied using survival rate by Kaplan-Meier method, univariate analysis by Log-rank test and generalized Wilcoxon test and multivariate analysis by proportional hazard model of Cox. The prognostic factors of pretreatment were age, serum Alb, serum LDH, pulmonary dysfunction of mixed type and performance status. In the classified study according to histology, age and pulmonary dysfunction of mixed type were prognostic importance in patients with squamous cell lung cancer. Hemoglobin in patients with squamous cell lung cancer was important, too. The chemotherapy given good response improved patient's prognosis.

Adenocarcinoma

Ultrastructural and cytochemical study of elastic fibers in the ventral aorta of a teleost, Anguilla japonica.

Previous studies have revealed that amorphous elastin and microfibrils are structural entities of mammalian elastic fibers. Elastin shows a wide phylogenetic distribution, but the presence of elastin-associated microfibrils has not been demonstrated in teleost aorta. Thus, we have ultrastructurally and cytochemically examined elastic fibers in the ventral aorta of eel, a teleost, by utilizing routine uranyl acetate and lead double staining, the tannic acid (pH 7.0)-uranyl acetate (TA-UA) method, elastase en bloc digestion, Thiéry's periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method, and the horseradish-peroxidase-labeled concanavalin A (Con A) method. In the ventral aorta of eel, a little ultrastructural difference between elastic fibers in the intima and media and those in the adventitia was noticed, but in either tunic each elastic fiber was basically composed of a "fibrillar core" and surrounding microfibrils. The fibrillar core was a collection of fibrils which showed a tendency to coalesce with each other, and these constituent fibrils were TA-UA positive and elastase-sensitive, representing their nature of elastin. By contrast, microfibrils associated with the fibrillar core were TA-UA negative and elastase-resistant, and their glycoproteinaceous nature was demonstrated by PA-TCH-SP and Con A methods. Thus, this study provides evidence for the presence of elastin-associated microfibrils in teleost aorta. These results are discussed in relation to the topographical difference of elastic fibers in eel aortic wall.

Anguilla

A new depsipeptide antibiotic, citropeptin.

An actinomycete identified as Streptomyces flavidovirens was found to produce a new cyclic hexadepsipeptide antibiotic, designated citropeptin. Citropeptin showed antitumor activity against P388 murine leukemia.

Animals

Ultrastructural cytochemistry of aortic microfibrils in the Arctic lamprey, Lampetra japonica.

In the ventral aorta of lamprey, microfibrils are major components of the extracellular matrix. With special reference to these microfibrils, we have cytochemically examined the lamprey ventral aorta, utilizing the tannic acid (pH 7.0)-uranyl acetate (TA-UA) method, elastase en bloc digestion, Thiéry's periodic acid-thiocarbohydrazide-silver proteinate (PA-TCH-SP) method, and ferritin- or horseradish peroxidase-labeled concanavalin A (Con A) methods. The lamprey microfibrils were strongly stained with PA-TCH-SP and both Con A methods, but did not show TA-UA staining nor elastase sensitivity. These cytochemical properties of lamprey microfibrils are identical with those of mammalian elastin-associated microfibrils. On the other hand, in spite of extensive examination, TA-UA positive and elastase-sensitive extracellular components were not found, so that lamprey ventral aorta does not appear to contain elastin. These results indicate that lamprey aortic connective tissue contains microfibrils as elastic components, but deposition of amorphous elastin does not occur.

Actin Cytoskeleton

Immunocytochemical demonstration of amelogenins and enamelins secreted by ameloblasts during the secretory and maturation stages.

Rabbit polyclonal antibodies against bovine amelogenins and enamelins which did not show any cross-reaction were raised, and ultrathin sections of rat incisors were examined by the protein A-gold and ABC methods. The immunoreactivity of amelogenins was found in dense granules in the intercellular spaces between preameloblasts, and later over the fine- and coarse-textured material. The immunoreactivity was present over the cell organelles associated with the secretory pathway, as well as pale and dark lysosomes of the presecretory and secretory ameloblasts. Here the enamel was immunolabeled in the intercrystal spaces. The immunoreactivity in multivesicular bodies was stronger in preameloblasts than in secretory ameloblasts. In the region of second ruffle-ended ameloblasts at the maturation stage, the immunolabeling was intense in the ruffled-border, but in the rough endoplasmic reticulum and Golgi apparatus, the immunolabeling was much weaker than at the secretory stage. The immunolabeling for enamelins showed essentially the same intracellular topographical pattern as that for amelogenins by the secretory stage, but was weaker. The immunoreactivity was found mainly attached to the enamel crystals. Double immunostaining of amelogenins and enamelins revealed that both immunoreactivities were present over the same cell organelles associated with secretion and lysosomal systems. It is suggested that the presecretory and secretory ameloblasts are actively involved in the secretion, degradation and resorption of enamel proteins, and that multivesicular bodies and lysosomes in the cells take part in these processes. Ameloblasts are considered to be related to the synthesis of enamelins.

Ameloblasts

Serum levels of pancreatic enzymes in lean and obese subjects.

Serum amylase and trypsin, but not lipase, were significantly lower in obese subjects than in lean subjects. Serum amylase and trypsin, but not lipase, had significant inverse correlation with body weight. Low serum amylase was associated with low protein, fat and carbohydrate intakes per kg body weight, whereas low serum trypsin was associated with low carbohydrate intake per kg body weight. Low serum pancreatic enzymes readily increased with diet therapy. It is assumed that lowered serum pancreatic enzymes in obese subjects are somehow related to their diet intake. The mechanism through which pancreatic enzymes in the blood reflect the composition of food is unclear.

Adult

Ultrastructural cytochemistry of oxytalan fibres in monkey periodontal ligaments with the high iron diamine method.

Monkey periodontal ligaments have been examined at the ultrastructural level to demonstrate the nature of reactive sites in oxytalan fibres. The high iron diamine (HID) and HID-thiocarbohydrazide-silver proteinate methods specific for sulphate groups, with and without prior oxidation with monopersulphate, were used. Oxytalan fibres were composed of bundles of microfibrils with a diameter of 11.5 +/- 1.7 nm (mean +/- S.D., n = 50). In cross section the microfibrils were found to have a denser periphery, giving them a 'tubular' appearance. The oxytalan microfibrils of non-oxidized specimens showed little reactivity with either HID method, except that the extracellular matrix material in close association with collagen fibrils stained weakly; in oxidized specimens, both HID methods strongly stained oxytalan microfibrils and weakly stained the extracellular matrix material. Such reactivity of oxytalan microfibrils was not altered by digestion with testicular hyaluronidase or chondroitinase ABC, performed prior to or after persulphate oxidation. Further, the sequential thiosulphation and HID method for the demonstration of disulphide and sulphhydryl groups stained oxytalan fibres moderately. These results indicate that the oxidative generation of sulphate groups in oxytalan fibres may occur from either disulphide or sulphhydryl groups, or both, rather than the result of unmasking of sulphated glycosaminoglycans.

Animals

Ultrastructure of extracellular polysaccharides produced by serotype c Streptococcus mutans.

The ultrastructure of extracellular polysaccharides produced in colonies by two clinical isolates and that of a nitrosoguanidine-induced mutant of serotype c Streptococcus mutans with different polysaccharide-synthesizing abilities were compared electron-microscopically. A large amount of polysaccharide was produced from sucrose by colonies of typical serotype c strain MT8148R and a clinical variant MT6801R with an enhanced fructan-synthesizing ability. Transmission electron-microscopy (TEM) revealed that the polysaccharides consisted of three structural components, i.e., globular, single-stranded filamentous, and double-stranded fibrillar structures. These structures were ascribed to production of fructan, water-soluble glucan, and water-insoluble glucan, respectively. On the other hand, two kinds of structures, a globular body and an amorphous substance, were observed by scanning electron-microscopy (SEM). The former was composed of fructan, while the latter contained a mixture of water-soluble and water-insoluble glucans which formed filamentous and double-stranded fibrillar structures under TEM. Very small quantities of polysaccharides were formed in colonies of mutant NG7183, which was derived from S. mutans MT6801R. This strain was found to possess low glucan- and no fructan-synthesizing abilities. The polysaccharides produced in colonies of mutant NG7183 were composed only of filamentous and double-stranded fibrils under TEM. A small amount of amorphous substance was observed by SEM in colonies of NG7183.

Dextranase